Download pdf - Session V Meat Processing and Packaging · 60 th International Congress of Meat Science and Technology, 17- 22 rd August 2014, Punta Del Este, Uruguay COMBINED EFFECTS OF HIGH PRESSURE

Session V Meat Processing and Packaging

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INVESTIGATION INTO CARCASS TRAITS AND SALEABLE MEAT YIELD OF AUSTRALIAN ALPACAS (VICUGNA PACOS)…………………………………………………..N/A

Smith, Melanie M. ; Bush, Russell D. ; Thomson, Peter C. ; Hopkins, David L.

COMBINED EFFECTS OF HIGH PRESSURE AND SODIUM HYDROGEN CARBONATE ON BEEF TEXTURE AND COLOR…………………………………………………………………………232

Nishiumi, Tadayuki ; Ohnuma, Shun ; Watanabe, Yui ; Sakata, Ryoichi

PHYSICOCHEMICAL CHARACTERISTICS OF LOW-FAT, LOW-SALT FRESH SHEEP

MEAT SAUSAGE………………………………………………………………………………………….236

Maia Junior, Jonhny A. ; Henry, Fábio C. ; do Valle, Felipe R. A. F. ; Ferreira, Karla S. ; Quirino, Célia R. ; Dutra de Resende, Eder

INSTRUMENTAL EVALUATION OF COLOR AND TEXTURE OF LOWFAT, LOW-SODIUM

CHLORIDE FRESH SHEEP MEAT SAUSAGE……………………………………………………..239

Maia Junior, Jonhny A. ; Henry, Fábio C. ; do Valle, Felipe R. A. F. ; Alvarenga Reis, Suelen ; Vilela Talma, Simone

EFFECT OF HIGH PRESSURE PROCESSING ON THE EATING QUALITY OF LOW FAT

SAUSAGE EMULSIONS………………………………………………………………………………..243

Yang, Huijuan ; Han, Minyi ; Bai, Yun ; Xu, Xinglian ; Zhou, Guanghong

ROLE OF LACTOBACILLUS SPP. AND STAPHYLOCOCCUS XYLOSUS IN MEAT: CONVERSION OF METMYOGLOBIN AND INHIBITION OF SPOILAGE BACTERIA…N/A

Kong, Baohua ; Li, Peijun ; Luo, Huiting

MOISTURIZING AGENTS AS DETERMINANTS OF PHYSICOCHEMICAL AND SENSORY CHARACTERISTICS OF DEER MEAT PRODUCTS……………………………………………..N/A

Sakata, Ryoichi ; Kaneko, Miyuki ; Lin, Liang-Chuan ; Wan, Tien-Chun

ENHANCED REDDENING OF MEAT BY THE ADDITION OF HIMALAYAN ROCK SALT…………………………………………………………………………………………………………247

Kaneko, Miyuki ; Okuda, Yasushi ; Waga, Masahiro ; Oshida, Toshio ; Sakata, Ryoichi

EFFECT OF MEAT AND NON-MEAT PROTEIN MIXING RATIO ON QUALITY CHARACTERISTIC OF EMULSION-TYPE SAUSAGE…………………………………………..250

Kang, Geunho ; Ham, Hyoung-Joo ; Cho, Soohyun ; Seong, Pil-Nam ; Kang, Sun-Mun ; Park, Kyoungmi ; Ba, Hoa Van ; Park, Beomyoung

THE POTENTIAL ROLE OF ARGININE IN COMPETITIVENESS AND FUNCTIONALITY OF COAGULASE-NEGATIVE STAPHYLOCOCCI DURING MEAT FERMENTATION……253

Leroy, Frédéric ; Sanchez-Mainar, Maria ; Weckx, Stefan ; De Vuyst, Luc

EFFECT OF MUSHROOM INCORPORATION ON QUALITY, SENSORY AND MICROBIAL STABILITY OF FRANKFURTER TYPE-SAUSAGE……………………………………………….257

Seong, Pil-Nam ; Park, Kyoungmi ; Cho, Soohyun ; Kang, Geunho ; Kang, Sun-Mun ; Park, Beomyoung ; Ba, Hoa Van

APPLICATION OF NEAR INFRARED SPECTROSCOPY FOR GRADING AND CLASSIFICATION OF 4 CHICKEN MEAT…………………………………………………………N/A

Fernandes Barbina, Douglas ; Barreto Honorato, Danielle Cristina ; Midori Kaminishikawahara, Cintia ; Lourenco Soares, Adriana ; Yurika Mizubuti, Ivone ; Grespan, Moises ; Shimokomaki, Massami ; Iouko Ida, Elza

REDUCTION OF CHILLING LOSSES IN BEEF CARCASSES BY WRAPPING WITH POLYETHYLENE STRETCH FILM……………………………………………………………………N/A

J. Lolatto, Danielly C. Neto, Angelo Polizel ; Roça, Roberto O. ; Moreira, Paulo S.A. ; da Rosa, Claudineli C.B. ; Dromboski, Tiago ; Costa, Leidiane M. ; Reis, Ellen C.A.

OPTIMISING THE SHELF-LIFE OF PORK CHOPS WITH NEW GAS COMPOSITIONS AND REDUCED HEADSPACE…………………………………………………………………………259

Bozec, A. ; Vautier, A. ; Lhommeau, T. ; Le Roux, A

LOSSES DUE AGING PROCESS IN BREAST MEAT OF BROILERS WITH DIFFERENT

GENDERS AND AGES…………………………………………………………………………………..263

M. Mello, Juliana L. ; Rodrigues, Ana Beatriz B. ; Giampietro-Ganeco, Aline ; Ferrari, Fábio B. ; Borba, Hirasilva ; de Souza, Rodrigo A. ; de Souza, Pedro A.

ATTRIBUTES RELATED TO TENDERNESS IN BREAST MEAT OF BROILERS WITH

DIFFERENT GENDERS AND AGES AFTER AGING……………………………………………..266

M. Mello, Juliana. L. ; Rodrigues, Ana Beatriz B. ; Giampietro-Ganeco, Aline ; Ferrari, Fábio B. ; Borba, Hirasilva ; Vieira, Leonardo D. C. ; de Souza, Rodrigo A.

EFFECT OF LACTATE AND DIACETATE SALTS AND HIGH PRESSURE PROCESSING ON

THE SURVIVAL OF LISTERIA MONOCYTOGENES IN CURED BEEF CARPACCIO……269

Barrio, Yanina X. ; Wilches Perez, Diego ; Vaudagna, Sergio R. ; Rovira, Jordi

ROOIBOS (ASPALATHUS LINEARIS) IS AN EFFECTIVE FUNCTIONAL INGREDIENT IN RETARDING LIPID OXIDATION OF COOKED RABBIT MEAT PATTIES……………273

Dalle Zotte, Antonella ; Mousavikhorshidi, Hannaneh ; Pizzocchero, Valentino ; Cullere, Marco

GREEN TEA EXTRACT ALTERS THE FUNCTIONAL PROPERTIES OF MEAT EMULSIONS BY GENERATION OF PROTEIN CROSS-LINKS………………………………………………..N/A

Jongberg, Sisse ; Therkelsen, Linda de S. ; Miklos, Rikke ; Lund, Marianne N.

HIGH HYDROSTATIC PRESSURE TREATMENT OF RAW MATERIAL: EFFECT ON PHYSICOCHEMICAL AND TEXTURE PROPERTIES OF BEEF PATTIES………………….N/A

Eccoña Sota, Amparo ; Sanow, Claudio ; Sancho, Ana M. ; Vaudagna, Sergio R.

QUALITY CHARACTERISTICS OF CHICKEN NUGGETS WITH REDUCED AMOUNTS OF SODIUM CHLORIDE AND ADDED DIETARY FIBER………………………………………….N/A

Bonato, P. ; Perlo, F. ; Fabre, R. ; Teira, G. ; Tisocco, O. ; Tito, B. ; Dalzotto, G.

EFFECT OF THE INCLUSION OF CHESTNUTS IN THE FINISHING DIET ON THE CHANGES IN CHEMICAL COMPOSITION DURING THE PROCESSING OF CELTA DRY-CURED HAM………………………………………………………………………………………………277

Bermudez, Roberto ; Franco, Daniel ; Carballo, Javier ; Carril, J.A. ; Lorenzo, José M.

MINIMISING PROTEIN OXIDATION IN RETAIL-PACKED MINCED BEEF USING

THREE-GAS MA-PACKAGING……………………………………………………………………….281

Spanos, Dimitrios ; Baussá, Laura ; Baron, Caroline P. ; Trngren, Mari Ann

OPTIMISING EATING QUALITY AND SHELF LIFE OF ENHANCED AND MARINATED PORK CHOPS USING THREE-GAS MAP………………………………………………………….285

Trngren, Mari Ann

SENSORY CHARACTERISTICS OF MURRAH BUFFALOES MEAT SUBMITTED TO

DIFFERENT PERIODS OF AGEING………………………………………………………………..N/A

Andrighetto, Cristiana ; da Luz, Patrícia A. C. ; Jorge, André M. ; Santos, Carolina T. ; Roça, Roberto O. ; Zanetti, Leonardo H. ; Sampaio Aranha, Aline

CHEMICAL AND MICROBIOLOGICAL CHARACTERISTICS OF MURRAH BUFFALOES MEAT SUBMITTED TO DIFFERENT PERIODS OF AGEING…………………………………N/A

Andrighetto, Cristiana ; da Luz, Patrícia A. C. ; Jorge, André M. ; Santos, Carolina T. ; Roça, Roberto O. ; Zanetti, Leonardo H. ; Domingues, Miguel S.

INFLUENCE OF FREEZING TIME ON THE WATER ACTIVITY AND LIPID OXIDATION IN MEAT OF BROILERS REARED AT THE CONVENTIONAL AND ANTIBIOTIC-FREE

SYSTEMS…………………………………………………………………………………………………..289

Giampietro-Ganeco, Aline ; Borba, Hirasilva ; Mello, Juliana L. M. ; Saccomani, Ana Paula de O. ; de Souza, Rodrigo A. ; de Souza, Pedro A.

DETERMINATION OF COOKING WEIGHT LOSS IN DIFFERENT TYPES OF BROILERS AND FREEZING PERIODS……………………………………………….................................291

Giampietro-Ganeco, Aline ; Borba, Hirasilva ; Mello, Juliana L. M. ; Junior, Luiz C. M. ; Rodrigues, Ana Beatriz B. ; Demarchi, Caroline ; Vieira, Leonardo D. C.

INFLUENCE OF PH ON (ZINC) PROTOPORPHYRIN IX FORMATION IN DRY FERMENTED SAUSAGES ASSESSED USING A FAST SCREENING METHOD………….N/A

De Maere, H. ; De Mey, E. ; Dewulf, L. ; Fraeye, I. ; Chollet, S. ; Paelinck, H.

PHYSICOCHEMICAL PARAMETERS AND SENSORY PROPERTIES OF LAMB BURGER MANUFACTURED WITH DIFFERENT CONCENTRATIONS OF OREGANO EXTRACT.............................................................................................................N/A

Paseto Fernandes, Rafaella de P. ; Trindade, Marco A. ; Tonin, Fernando G. ; P. Pugine, Silvana M. ; Hirano, Merícia H. ; Lorenzo, José M. ; de Melo, Mariza P.

EFFECTS OF PORK ROUND POWDER ADDITION ON THE QUALITY CHARACTERISTICS OF NOODLE PRODUCT…………………………………………………….294

Jeon, Kihong ; Kim, Youngboong ; Kim, Youngho ; Hwang, Yoonseon ; Choi, Jinyoung

EFFECT OF OXYGEN ON THE OXIDATION OF TWO DIFFERENT MAP RETAIL PORK

MEAT PRODUCTS……………………………………………………………………………………….N/A

Spanos, Dimitrios ; Trngren, Mari Ann ; Christensen, Mette ; Baron, Caroline P.

DEVELOPMENT OF RAMAN SPECTROMETRIC ON-SITE METHOD TO MEASURE SOLID FAT CONTENT……………………………………………………………………………………………N/A

Motoyama, Michiyo ; Hayashi, Shigenobu ; Jimura, Keiko ; Chikuni, Koichi ; Nakajima, Ikuyo ; Aikawa, Katsuhiro ; Sasaki, Keisuke

RAPID AUTHENTICATION OF ENHANCED QUALITY PORK BY VISIBLE AND NEAR INFRARED SPECTROSCOPY…………………..…………………………………………………....299

Prieto, N. ; Prieto, N. ; Juárez, M. ; Zijlstra, R. T. ; López-Campos, Ó. ; Aalhus, J. L.

INFLUENCE OF MODIFIED ATMOSPHERE PACKAGING ON PHYSICAL-CHEMICAL PARAMETERS OF LOIN LAMB STORED UNDER REFRIGERATION………………………N/A

Rodrigues dos Santos, Isabela ; Souza Palú, Ana Flávia ; Santa Barbara, Beatriz Zillo ; Augusto Gallo, Fábio ; A. Freire, Maria Teresa ; TRINDADE, M.A.

TECHNOLOGICAL RESPONSE OF TWO BOVINE MUSCLES SUBJECTED TO FREEZE-

DRYING PROCESS……………………………………………………………………………………..N/A

Messina, Valeria ; Paschetta, Fernanda ; Chamorro, Verónica C. ; Sancho, Ana M. ; de Reca, Noemí Walsöe ; Pazos, Adriana A. ; Grigioni, Gabriela

EFFECT OF SODIUM, POTASSIUM AND CALCIUM CHLORIDE ON PROTEOLYSIS IN DRY FERMENTED SAUSAGES……………………………………………………………………….N/A

Dos Santos, B. A. ; Dos Santos, B. A. ; Campagnol, P. C. B. ; Wagner, R. ; Pollonio, M.A.R.

STABILITY OF LAMB BURGER CONTAINING NATURALANTIOXIDANT EXTRACT: OXIDATIVE AND COLOUR PARAMETERSDURING TWO MONTHS OF FROZEN

STORAGE………………………………………………………………………………………………….N/A

de P. Paseto Fernandes, Rafaella ; Trindade, Marco A. ; Tonin, Fernando G. ; P. Pugine, Silvana M. ; Hirano, Merícia H. ; Lorenzo, José M. ; de Melo, Mariza P.

EFFECTS OF ELECTRICAL STIMULATION TO LOCALIZED PARTOF CARCASSES ON SHEAR FORCE OF HANWOO BEEF………………………………………………………………..N/A

Yang, Jieun ; Jeong, Dawoon ; Amna, Touseef ; Park, Beom-Young ; Cho, Soohyun ; Hwang, Inho

DIELECTRIC PROPERTIES OF SELECTED PORK ORGAN MEATSAND EXPERIMENTAL PATE PRODUCT FORMULATIONS FORCONTINUOUS FLOW MICROWAVE

PROCESSING…………………………………………………………………………………………….303

Hanson, Dana J. ; Kemal, Mehmet ; Tennant, Travis ; Simunovic, Josip

THE EFFECT OF BRINE INJECTION LEVEL ON MOISTURE RETENTION AND SENSORY PROPERTIES OF CHICKEN BREAST MEAT………………………………………………………308

Moholisa, E. ; Roodt, E. ; Bothma, C. ; de Witt, F. ; Hugo, A.

MONITORING THE LIPID STRUCTURE OF FRANKFURTERS: TOWARD A FOODMODEL TO BETTER UNDERSTAND THE PERCEPTION OF FAT………………………………………N/A

Chapeau, A-L. ; Promeyrat, A. ; Astruc, Thierry ; Vénien, A. ; Ferreira, C. ; Santé-Lhoutellier, V

GENETIC POLYMORPHISM OF RYR 1 GENE FROM TURKEY PSE BREAST MUSCLE…………………………………………………………………………………………………….312

Gonzales Paião, Fernanda ; Ribeirete, Viviane ; de Carvalho, Rafael Humberto ; Pedrão, Mayka Reghiany ; Garcia Coró, Fábio Augusto ; Shimokomaki, Massami

ANTIOXIDANT AND ANTIMICROBIAL POTENTIAL OF JABUTICABA (MYRCIARIA CAULIFLORA) EXTRACT ADDED TO FRESH SAUSAGES……………………………………N/A

Baldin, Juliana C. ; MUNEKATA, P. E. S. ; Canan, Thais M. ; Michelin, Euder C. ; Flipsen, Keila D. ; FREGONESI, R. P. ; S. Godoy, Silvia H. ; F. Trindade, Carmen S. ; Fernandes, Andrezza M. ; TRINDADE, M.A.

USE OF ACID TURKEY BREAST MEAT IN EMBEDED PRODUCTS: FORMULATION IMPACTS ON TECHNOLOGICAL CHARACTERISTICS………………………………………..N/A

Kondo, Henry ; Gonzales Paião, Fernanda ; Gallo, André ; Augusto Iziquiel, Cézar ; Aparecida Mafra, Magali; Pedrão, Mayka Reghiany

HEADCHEESE AS AN ALTERNATIVE USE OF PORK BYPRODUCTS: PHYSICOCHEMICAL ANALYSIS AND TEXTURE PROFILE………………………………….N/A

Silva Junior, Jamil ; Ribeiro, Elissama R. ; Pedrão, Mayka Reghiany ; Machado-Lunkes, Alessandra

RESTRUCTURED AND BREADED (NUGGETS) CHICKEN PRODUCT WITH THE ADDITION OF OAT FIBER: CHEMICAL AND SENSORIAL CHARACTERIZATION …315

Fiorentin, Cristiane ; Pedrão, Mayka Reghiany ; Gallo, André ; Augusto Iziquiel, Cézar ; Aparecida Mafra, Magali ; Garcia Coró, Fábio Augusto

N/A= The short paper is not available, either because the authors reported to the editor they do

not wish to publish their work, or because the authors did not respond to the editor's invitation

to publish.

60th International Congress of Meat Science and Technology, 17-22rd August 2014, Punta Del Este, Uruguay

COMBINED EFFECTS OF HIGH PRESSURE AND SODIUM HYDROGEN CARBONATE ON BEEF TEXTURE AND COLOR

Tadayuki Nishiumi1*, Shun Ohnuma1, Yui Watanabe1 and Ryoichi Sakata2 1 Division of Life and Food Science, Graduate School of Science and Technology, Niigata University, Niigata, Japan

2 School of Veterinary Medicine, Azabu University, Sagamihara, Japan

* [email protected]

Abstract - Effect of combined high-pressure and sodium hydrogen carbonate (NaHCO3) treatment on the physical properties and color of silverside Australian beef was investigated. Meat samples were pressurized at 100 to 500 MPa and the water content, weight reduction, rupture stress, and meat color were determined. The water content of meat treated with NaHCO3 and high pressure (300 MPa) reached a maximum of 70.1%. Weight reduction tended to decrease with high-pressure treatment at 300 MPa. Meats treated with NaHCO3 and high pressure at 400 MPa showed a more than 50% decrease in toughness. Whitening of the meat was reduced by the combined high-pressure and NaHCO3 treatment, which had been maintained during refrigeration storage. Therefore the combined high-pressure and NaHCO3 treatment is effective for improvement of Australian beef quality. I. INTRODUCTION There are a number of reports on the effects of high pressure processing of meat and meat products. In a previous study of high-pressure treatment of meat, Ananth et al. (1) reported that color, peak load, water-holding capacity, and moisture were not different between samples treated with high pressure (414 MPa) for 9 min at 25°C and controls with cooking. In contrast, Souza et al. (2) reported improvements in the water-holding capacity and pork palatability by high-pressure treatment. High-pressure treatment can accelerate meat conditioning and, as a result, induce a great tenderization of raw meat (3). However, its effectiveness is limited on cooking. Many researchers have reported that myoglobin, which is associated with the red

pigmentation of meat, is denatured by high-pressure treatment. Shigehisa et al. (4) observed a decrease in the L* and a* values of pork slurries pressurized at 100 to 600 MPa. Carlez et al. (5) reported that the color of minced beef changed markedly, especially above 300 MPa. They also reported that meat discoloration from pressure processing may result from a ‘whitening effect’ in the range of 200 to 350 MPa due to globin denaturation and heme displacement or release, or protein coagulation with a resulting loss of solubility of sarcoplasmic and/or myofibrillar proteins that affect structure and surface properties (6). Sodium hydrogen carbonate (NaHCO3) treatment has been used traditionally in cooking as a meat tenderizer. However, NaHCO3 treatment causes cavity formation between the muscle fibers after heating. Kim et al. (7) applied a combined high-pressure and NaHCO3 treatment for pork and showed improved tenderness and juiciness without any cavity formation after heating. In the present study, we investigated the effects of combined high-pressure and NaHCO3 treatment on the physical properties and color of beef which is firm and difficult to chew. II. MATERIALS AND METHODS Sample preparation Silverside Australian beef was used as the meat sample, this cut being a part of the outside round and the most tough in beef. The samples were purchased from Itoham Foods Inc. and stored at -20°C. The meat was thawed at 4°C overnight. The meat was cut along the grain into slices 1 cm in thickness. The meat samples were then placed individually in vacuum-sealed polyethylene bags with 4 vol (v/w) of 0.4 M NaHCO3 solution or de-ionized water as the control.

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The samples were sealed and placed in a 20°C water bath for 40 min. They were then removed from the soaking solution and each meat sample was again vacuum-sealed in a polyethylene bag. Meat samples were pressurized at 0.1, 100, 200, 300, 400, and 500 MPa at 20°C for 10 min using a high-pressure food processor (Dr. CHEF, Kobe Steel, Japan). Non-pressurized meat was expressed as the meat treated under 0.1 MPa. After pressurization, each meat sample was heated for 30 min at 80°C and cooled down in ice-cold water until the core temperature of the meat reached 20°C. Measurement of water content and weight reduction Water content was measured using a halogen moisture analyzer (HG63, Mettler Toledo, Switzerland). Approximately 3 g of each meat sample was heated at 137°C. For determination of weight reduction, the weight of the meat was measured after thawing, and re-measured after heating. Weight reduction was shown in ratio (%) for the meat weight after thawing. Measurement of rupture stress The rupture stress of each heated meat sample was measured using a rheometer (Creep Meter RE2-3305B, Yamaden, Japan). Samples were cut into pieces 25 × 20 × 10 mm in size and punctured by using a plunger (wedge shape, 8 × 0.25 mm, 20-mm long) at 10 mm/s, stopping at 100% of the thickness, using a load cell of 200 N. The rupture stress experiment was conducted at room temperature (20°C). Measurement of meat color Meat color was assessed during refrigeration storage under an atmospheric condition after high-pressure treatment. The L* (lightness), a* (redness), and b* (yellowness) values were determined using a chroma meter (CR-400, Konica Minolta, Japan) in the 1976 CIELAB system. Surface determinations were carried out on each sample. Statistical analysis Results are expressed as the mean ± SEM. All data were analyzed using one-way analysis of variance. The statistical significance of differences among means was

evaluated using Student’s t-test at the 1% level. III. RESULTS AND DISCUSSION Water content and weight reduction The effects of high-pressure treatment on water content and weight reduction of meat were compared between control and NaHCO3 treatment groups (Fig. 1). Water content of NaHCO3 treated meats (68.0±1.0%) increased by approximately 6.5% in comparison with untreated meats (61.5±1.1%) (p<0.01). Significant differences were not observed in the meats treated without NaHCO3, regardless of the processing pressure. The effect of high-pressure treatment was obvious with 300 MPa and 400 MPa pressure treatments (p<0.01) in NaHCO3 treated meats. Specifically, the water content of meats treated with NaHCO3 and high pressure at 300 MPa (70.1±0.6%) increased approximately 8.6% in comparison with untreated meats (61.5±1.1%). In isolated myofibrils from rabbit skeletal muscle exposed to pressure at 200 MPa, the structural continuity of the sarcomere was completely lost with the loss of Z-line (8). The increase in water capacity in this study may be due to the high-pressure rupture of the myofibril ultrastructure. The weight reduction of NaHCO3 treated meats (22.6±1.9%) was approximately 16.8% less than of the untreated meats (39.4±0.3%) (p<0.01). Significant differences were not observed in either untreated or NaHCO3 treated meats, regardless of the given pressure. However, weight reduction tended to decrease at 300 MPa. It is suggested that a combined high-pressure and NaHCO3 treatment is effective for limiting the loss of water content and weight during the cooking of beef. Rupture stress The effect of high-pressure treatment on rupture stress in untreated and NaHCO3 treated samples is shown in Fig. 2. The rupture stress of untreated and NaHCO3

treated meats was 3.97 × 106 N/m2 and 2.89 × 106 N/m2, respectively. The values significantly decreased with high-pressure treatment above 100 MPa in untreated

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samples and above 200 MPa in NaHCO3 treated samples (p<0.01). In particular, meats treated with NaHCO3 and high pressure at 400 MPa (1.74 × 106 N/m2) showed a more than 50% decrease in hardness compared to untreated meats (3.97 × 106 N/m2). Kim et al. (7) reported that pressurization at 400 MPa showed the greatest effect on pork ham tenderness. In this study, the combined high-pressure and NaHCO3 treatment could improve the tenderness of beef as shown in pork. Meat color The effect of high-pressure treatment on the meat color of untreated and NaHCO3 treated samples are shown in Table 1. The L* (lightness) value of NaHCO3 treated meats (30.41±2.00) was approximately 8.83 units lower than untreated meats (39.24±2.98) (p<0.01). The L* value of meats treated with high pressure (more than 300 MPa) significantly increased (p<0.01). The b* (yellowness) value of NaHCO3 treated meats (4.21±1.26) was also approximately 3.89 units lower than untreated meats (8.10±1.10) (p<0.01). As similar to the L* value result, the b* value of meats treated with high pressure (more than 300 MPa) significantly increased (p<0.01). The significant increases observed in L* and b* values were thought to be due to the whitening of the meat. The trend a* (redness) value is characteristically different from that of L* and b* values. The a* value of NaHCO3

treated meats (11.93±1.43) was approximately 1.92 units higher than untreated meats (10.01±0.75) (p<0.01). The a* value of meats treated with high pressure at 100 MPa and 200 MPa increased in comparison with untreated meats. However, a* value of meat subjected to high pressure more than 400 MPa decreased in comparison with that subjected to 300 MPa (12.28±1.34). This is considered to be due to myoglobin denaturation by high-pressure treatment (5). Since a* value of meat treated with NaHCO3 and high pressure (more than 400 MPa) increased, it is suggested that NaHCO3 treatment affects the structure of myoglobin and the effect on color is depending on the degree of pressure. Therefore, in the color of silverside Australian beef, a darkening was induced by

NaHCO3 treatment, while the whitening effect was induced by high pressure at 300 MPa or more. On the other hand, by using a combined high-pressure and NaHCO3 treatment, an inhibition of the increase of the L* value was induced with an increase of the a* value, which is desirable for consumers. The color of the meat processed with high pressure and NaHCO3 was maintained during refrigeration storage, for approximately two weeks.

Fig. 1. Effect of high-pressure and NaHCO3 treatment on (a) water content and (b) weigh reduction of beef. -●-: untreated, -○-: NaHCO3 treatment. Values are expressed as mean ± standard deviation, n=5.

Fig. 2. Effect of high-pressure and NaHCO3 treatment on the rupture stress of beef. -●-: untreated, -○-: NaHCO3 treatment. Values are expressed as mean ± standard deviation, n=22.

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Table 1. Effect of high-pressure and NaHCO3 treatment on the beef color.

Values are expressed as mean ± standard deviation, n=20. Significant differences among values in the same row are indicated by different letters (p<0.01).

IV. CONCLUSION The combination of high-pressure at 300-400 MPa and 0.4 M sodium hydrogen carbonate treatment was shown to be effective for the improvement of Australian beef quality, resulting from several physical properties and color of the meat. ACKNOWLEDGEMENTS This work was supported in part by the Niigata Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence from Japan Science and Technology Agency. REFERENCES 1. Ananth, V., Dickson, J. S., Olson, D. G. &

Murano, E.A. (1998). Shelf life extension, safety and quality of fresh pork loin treated

with high hydrostatic pressure. Journal of Food Protection 61: 1649-1656.

2. Souza, C. M., Boler, D. D., Clark, D. L., Kutzler, L. W., Holmer, S. F., Summerfield, J. W., Cannon, J. E., Smit, N. R., Mckeith, F. K. & Killefer, J. (2011). The effects of high pressure processing on pork quality, palatability, and further processed products. Meat Science 87: 419-427.

3. Suzuki, A. (1995). Mechanism for meat tenderization and acceleration of meat conditioning induced by high pressure treatment. Nippon Shokuhin Kagaku Kogaku Kaishi 42: 388-394.

4. Shigehisa, T., Ohmori, T., Saito, A., Taji, S. & Hayashi, R. (1991). Effects of high hydrostatic pressure on characteristics of pork slurries and inactivation of microorganisms associated with meat and meat products, International Journal of Food Microbiology 12: 207-215.

5. Carlez, A., Veciana-Nogues, T. & Cheftel, J. C. (1995). Changes in colour and myoglobin of minced beef meat due to high pressure processing, LWT - Food Science and Technology 28: 528-538.

6. Goutefongea, R., Rampon, V., Nicolas, J. & Dumont, J. P. (1995). Meat color changes under high pressure treatment. In Proceedings of the 41st International Congress of Meat Science and Technology (pp. 384-385), 20-25 August 1995, San Antonio, U.S.A.

7. Kim, Y. -J., Nishiumi, T., Fujimura, S., Ogoshi, H. & Suzuki, A. (2013). Combined effects of high pressure and sodium hydrogen carbonate treatment on pork ham: improvement of texture and palatability. High Pressure Research 33: 354-361.

8. Suzuki, A., Suzuki, N., Ikeuchi, Y. & Saito M. (1991). Effect of high pressure treatment on the ultrastructure and solubilization of isolated myofibrils, Agricultural and Biological Chemistry 55: 2467-2473.

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PHYSICOCHEMICAL CHARACTERISTICS OF LOW-FAT, LOW-SALT FRESH SHEEP MEAT SAUSAGE

Jonhny A. Maia Junior1,2*; Fábio C. Henry1; Felipe R. A. F. do Valle1; Karla S. Ferreira1; Célia R.

Quirino1; Eder Dutra de Resende1

1 Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, Brasil 2 Prefeitura Municipal de Macaé, Brasil

*[email protected]

Abstract – This study evaluated the centesimal composition of fresh sheep meat sausages prepared with passion fruit meal and potassium chloride (KCl). Seven formulations were prepared, six containing different percentages of passion fruit meal and KCl, and one control. Centesimal composition analysis showed that formulations F1, F2, and F3 had low fat contents, which were reduced by 59% with the addition of passion fruit meal, compared with the control. No significant difference was observed in sodium levels, probably due to the presence of sodium nitrite and sodium erythorbate in formulations. The formulations are in accordance with the quality standards defined for the fresh sausages. Formulations F1, F2 and F3 presented a reduction of approximately 40% in fat content, and F2 and F3 had a non-significant decrease in sodium levels. F3 showed the best results. I. INTRODUCTION Fresh sausage is a raw meat product containing pork fat and other ingredients to improve properties like texture, conservation and flavor. However, these ingredients have high sodium and fat levels (1, 2, 3, 4). Sodium, calorie and alcohol excess as well as deficiency in calcium and potassium are the main nutrition factors associated with systemic arterial hypertension (SAH), as well as the interaction between genetic and environmental variables (5, 6). SAH prevalence is high in the Brazilian population, and affects approximately 10% to 45% of the country’s adult population (7, 8, 6) and between 1% and 2% of children and teenagers (9). The excessive consumption of fat may cause health problems such as arteriosclerosis, colon cancer, obesity and others. Therefore, consumers today prefer low-fat foods that retain the sensory characteristics of more traditional products (10). The present study evaluated the influence of passion fruit meal and potassium chloride (KCl) in the

physicochemical characteristics of fresh sheep meat sausages. II. MATERIAL AND METHODS Sheep shoulder blade cuts and pork fat were used in sausage formulations. All ingredients were purchased in local shops and appropriately inspected. Passion fruit meal was prepared in the Laboratory of Food Technology, (LTA, UENF). Experiments were carried out in Federal Institute Espírito Santo (IFES), in Alegre, state of Espírito Santo (ES), and in LTA-UENF. Seven formulations were prepared with fresh sheep meat, one of which was used as control. All formulations were defined after five pretests, which established acceptable levels of fat and sodium. Table 1 shows the decrease in fat and sodium in each formulation. Table 1. Reduction of fat and NaCl levels in fresh sheep

meat sausages (%). Formulation Fat NaCl Control 0 0 F1 59 0 F2 59 50 F3 59 25 F4 31 0 F5 31 50 F6 31 25

Fat was partially replaced by passion fruit meal, according to Oliveira (11), containing on average 26.4% pectin. Reduction of sodium levels was carried out replacing NaCl by KCl. Formulations are presented in Table 2.

Table 2. Composition of the seven fresh sheep meat

sausage formulations (%) Raw material Control F1 F2 F3 F4 F5 F6

236

Sheep meat 76.66 82.36

82.36

82.36

80.30

80.30

80.30

Fat 20.00 8.20

8.20

8.20

13.80

13.80

13.80

Salt (NaCl) 2.20 2.20

1.10

1.65

2.20

1.10

1.65

KCl 0.00 0.00

1.10

0.55

0.00

1.10

0.55

Sugar 0.095 0.095

0.095

0.095

0.095

0.095

0.095

Water 1.00 4.70

4.70

4.70

2.35

2.35

2.35

Passion fruit meal 0.00

2.40

2.40

2.40

1.20

1.20

1.20

Sodium nitrite

0.015 0.015

0.015

0.015

0.015

0.015

0.015

Sodium erythorbate

0.025 0.025

0.025

0.025

0.025

0.025

0.025

The data obtained were submitted to Analysis of Variance (ANOVA) and the Student-Newman-Keuls (SNK) test in the software SAS version 9.3 (12). III. RESULTS AND DISCUSSION Table 3 shows the results of the analyses fresh sheep meat sausage composition.

Table 3. Means (%) and standard deviations (SD) of centesimal compositions of sausage formulations and

quality standards for fresh sheep meat sausage composition.

Centesimal composition of the seven formulations of raw sheep meat sausage was in accordance with the quality standards for the product, defined by the Ministry of Agriculture, Livestock and Food Supply (1). The results for formulations F1, F2 e F3 did not differ in fat levels (p > 0.05). The same was observed for F4 and F5, and F4 and F6. In the present study, mean fat level of formulations F1, F2

and F3 was approximately 40% lower, and the decrease in fat/100 g solids was greater than 3 g. These formulations are categorized as light products, that is, those that present low fat levels, compared with conventional products, as defined by Resolution 27, Health Ministry, from 13/1/1998 (13), replaced by Resolution 54, from 12/11/2012 (14). The formulations are according to the quality standards for fresh meat sausages, which establishes a maximum of 30% fat (1). Tobin et al. (15) analyzed 20 samples of frankfurters produced in Ireland containing different salt and fat levels. Fat levels varied between 9.61% and 25.42% (mean: 17.52%), which differed from those reported in the present study. Tobin et al. (16) analyzed 28 samples of pork breakfast sausages also produced in Ireland, with variable contents of salt and fat. The values observed ranged between 22.27% and 37.98% (mean: 30.00%). These values also differed from those obtained in the present study. These differences may be due to the raw material used and fat levels, which were higher than those used here. Regarding sodium levels no difference was detected (p > 0.05) between formulations, due to the use of sodium nitrite and sodium erythorbate. The replacement of NaCl by KCl did not lead to a decrease in sodium levels. Carraro et al. (17) reported that sodium levels did not fall in mortadella, due to the presence of sodium erythorbate and sodium polyphosphate in formulations, confirming the values obtained in the present study. However, Corral et al. (18) observed a decrease in sodium levels in fermented encased meat products. IV. CONCLUSION The formulations prepared in the present study met the quality standards defined for fresh sausages. Formulations F1, F2 and F3 met the requirements in Resolution 54/2012, Brazilian Health Ministry, which specifies low-fat products. Formulation F3 afforded the best results in the present study. However, further studies are necessary to assess the sensorial acceptance of this product. ACKNOWLEDGEMENTS The authors thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the grants given, Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) for financial support, and the City Administration of Macaé for the support to carry out this study.

Raw material Fat Protein Water Na g/100g

Control 18.03a ±

1.24 16.30a ±

0.33 59.86d ±

0.84 889.14a ±

114.09

F1 10.12d ±

0.65 17.04a ±

0.71 66.19a ±

0.43 908.43a ±

48.00

F2 10.55d ±

0.70 16.78a ±

1.23 65.01a ±

0.60 812.20a ±

134.40

F3 10.33d ±

0.29 17.01a ±

0.90 65.47a ±

0.34 797.54a ±

80.00

F4 13.50bc ±

0.77 16.07a ±

1.86 63.24b ±

0.75 831.86a ±

107.62

F5 14.61b ±

0.65 17.01a ±

0.75 61.09c ±

0.95 901.92a ±

124.34

F6 12.92c ±

0.56 16.10a ±

1.30 62.27b ±

0.61 886.02a ±

115.57 Quality standard

≤ 30.00 ≥ 12.00 ≤ 70.00

237

REFERENCES 1. Brasil. (2000). Ministério da Agricultura e Abastecimento. Instrução normativa n. 4, de 31 de março de 2000. Regulamentos técnicos de identidade e qualidade de carne mecanicamente separada, de mortadela, de linguiça e de salsicha. Diário Oficial da Republica Federativa do Brasil, Brasília, 05 abr. 2000, Seção 1, p. 6. 2. Ferrari, C. C., Soares, L. M. V. (2003). Concentrações de sódio em bebidas carbonatadas nacionais. Revista Ciência Tecnologia de Alimentos 23: 414-417. 3. Almeida, O. C. (2005). Avaliação físico-química e microbiológica de linguiça toscana porcionada e armazenada em diferentes embalagens, sob condições de estocagem similares às praticadas em supermercados. 2005, 80p. Dissertation (MSc, Food Technologies), Food Engineering School, University of de Campinas (UNICAMP), Campinas. 4. Raimundo, A., Couto, S. M., Lanzillotti, H. S. (2005). Elaboração e análise sensorial de linguiças caseiras. Revista Higiene Alimentar 128: 70-77. 5. Cabral, P. C., Melo, A. M. C. A., Amado, T. C. F., Santos, R. M. A. B. (2003). Avaliação antropométrica e dietética de hipertensos atendidos em ambulatório de um hospital universitário. Revista de Nutrição 16: 61-71. 6. Molina, M. D. C. B., Cunhab, R. S., Herkenhoffb, L. F., Millb, J. G. (2003). Hipertensão arterial e consumo de sal em população urbana. Revista Saúde Pública, 37: 743-750. 7. Reis, N. T., Cople, C. S. (1999). Nutrição clínica na hipertensão arterial. Rio de Janeiro: Revinter. 8. Castro, I., Batlouni, M., Cantarelli, E., Ramires, J. A. F., Luna, R. L., Feitosa, G. S. (1999). Cardiologia: princípios e prática. Porto Alegre: Artes Médicas. 9. Kater, C. E., Costa-Santos, M. (2001). O espectro das síndromes de hipertensão esteróide na infância e adolescência. Arquivos Brasileiros de Endocrinologia e Metabolismo 45: 76-86. 10. Galvan, A. P., Rosa, G., Back, J., Lima, D. P., Corso, M. P. (2011). Desenvolvimento de linguiça tipo Toscana com teor reduzido de gordura e adição de pectina e inulina. In Encontro Paranaense de Engenharia de Alimentos, 3, 2011. Guarapuava/PR. 11. Oliveira, E. M. S. (2009). Caracterização de rendimento das sementes e do albedo do maracujá para aproveitamento industrial e obtenção da farinha da casca e pectina. 2009. Dissertion. (MSc Plant Production). Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes. 12. SAS. (2009). User’s guide statistics. Cary: Institute SAS. 13. Brasil (1998) BRASIL. Portaria n. 27 SVS/MS, de 13 de janeiro de 1998. Regulamento Técnico Referente à Informação Nutricional Complementar. Dário Oficial da União, 16 jan. 1998. Seção 1. Available at <http://www.in.gov.br.>. Accessed on 17/12/2013. 14. Brasil, Agência Nacional de Vigilância Sanitária. (2003). RDC no 54, de 12 de novembro de 2012.

Available at <http://www.anvisa.gov.br>. Accessed on 27/12/2013. 15. Tobin, B. D., O’Sullivan, M. G., Hamill, R. M., Kerry, J. P. (2012). Effect of varying salt and fat levels on the sensory and physiochemical quality of frankfurters. Meat Science. 92: 659-666. 16. Tobin, B. D., O’Sullivan, M. G., Hamill, R. M., Kerry, J. P. (2013). The impact of salt and fat level variation on the physiochemical properties and sensory quality of pork breakfast sausages. Meat Science. 93:145-152. 17. Carraro, C. I., Machado, R., Espindola, V., Campagnol, P. C. B., Pollonio, M. A. R. (2012). The effects of sodium reductions and the use of herbs and spices on the quality and safety of bologna sausage. Ciência e Tecnologia de Alimentos 32. Available at: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612012000200013. Accessed on 14/01/2014. 18. Corral, S., Salvador, A., Flores, M. (2013). Salt reduction in slow fermented sausages affects the generation of aroma active compounds. Meat Science 93: 776-785.

238

http://www.in.gov.br/

http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612012000200013

http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612012000200013

INSTRUMENTAL EVALUATION OF COLOR AND TEXTURE OF LOW-FAT, LOW-SODIUM CHLORIDE FRESH SHEEP MEAT SAUSAGE

Jonhny A. Maia Junior1,2*; Fábio C. Henry1; Felipe R. A. F. do Valle1; Suelen Alvarenga Reis1;

Simone Vilela Talma1,3

1 Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, Brasil 2 Prefeitura Municipal de Macaé, Brasil

3 Instituto Federal de Sergipe, Campus Glória, Brasil

*[email protected]

Abstract – This study evaluated instrumental measurements of color and texture of fresh sheep meat sausages containing passion fruit meal (PM) and KCl. Color analysis revealed no significant difference (p > 0.05) in L* values between F1, F2, F3 and F4 or between F4 and F5 and C, F5 and F6. Mean a* values in formulation C were different from the others (p < 0.05). No significant difference was observed (p > 0.05) between F4, F5 and F6, as well as F1, F2, F3, F4 and F5. Mean texture varied between formulations. No significant difference was observed (p > 0.05) in hardness between F2, F4, F5 and F6, or between F1, F3, F4, F5 and F6 and C. Mean chewiness also varied across formulations. No significant difference was observed (p > 0.05) between F1, F2, F4, F5 and F6, and between F1, F3, F4 and F6 and C, F1 and F3. The results show that formulations in which fat levels were the lowest because of replacement with PM, especially F2, presented paler red color and higher resistance, possibly due to the stabilizing and thickening properties of the pectin in PM. I. INTRODUCTION The awareness of the importance of healthy food habits, the concerns with quality of life and the commercialization of diet products have led more and more consumers to prefer diet and light food products. Additionally, the high prevalence of diabetes mellitus, cardiovascular diseases, hypertension and obesity also plays a role in this growth. In this context, one of the main concerns in the food industry is the development of products that meet the requirements of different consumer profiles. Among these, the group of people with hypertension stands out, due to low sodium restrictions in diets (1). Epidemiological studies have shown that individuals consuming low sodium diets presented lower blood pressure levels, compared with those that consumed sodium-free foods (2). In Brazil, studies have investigated the effect of the reduction of sodium levels on sensorial properties

of Toscana sausages (3), of the reduction of fat and salt contents in pork sausages using xantham gum and potassium chloride (KCl) (4), and of the reduction of fat content in Toscana sausage using pectin and inulin (5). Apart from the products mentioned, low-fat, low-salt fresh sheep meat sausages are considered potentially healthier foods. High fiber and potassium foods are believed to lower the risk of systemic hypertension, and are seen as “protection foods” (6). In turn, low-fat, low-sodium encased meat products have become the object of several studies. In this sense, this study aimed at assessing instrumental color and texture of fresh sheep meat sausages containing passion fruit meal (PM) and KCl. II. MATERIALS AND METHODS Samples of fresh sheep meat sausages were prepared according to the recommendations defined by the National Institute of Industrial Property (INPI) in the reference agroindustrial plant administered by Federal Institute Espírito Santo (IFES), in Alegre, state of Espírito Santo (ES), Brazil. Seven formulations were used, one of which was the control. Formulations were chosen after five pretests, which defined acceptable levels of reduction of fat and sodium levels. Table 1 shows the decrease in fat and sodium in each formulation.

Table 1. Reduction of fat and NaCl levels in fresh sheep

meat sausages (%). Formulation Fat NaCl Control 0 0 F1 59 0 F2 59 50 F3 59 25 F4 31 0 F5 31 50 F6 31 25

239

Fat was partially replaced by passion fruit meal, according to Oliveira (7), containing on average 26.4% pectin. Reduction of sodium levels was carried out replacing NaCl by KCl. Formulations are presented in Table 2.

Table 2. Composition of the seven fresh sheep meat sausage formulations (%)

Raw material

Control F1 F2 F3 F4 F5 F6

Sheep meat 76.66 82.36

82.36

82.36

80.30

80.30

80.30

Fat 20.00 8.20

8.20

8.20

13.80

13.80

13.80

Salt (NaCl) 2.20 2.20

1.10

1.65

2.20

1.10

1.65

KCl 0.00 0.00

1.10

0.55

0.00

1.10

0.55

Sugar 0.095 0.095

0.095

0.095

0.095

0.095

0.095

Water 1.00 4.70

4.70

4.70

2.35

2.35

2.35

Passion fruit meal 0.00

2.40

2.40

2.40

1.20

1.20

1.20

Sodium nitrite

0.015 0.015

0.015

0.015

0.015

0.015

0.015

Sodium erythorbate

0.025 0.025

0.025

0.025

0.025

0.025

0.025

Samples were wrapped in aluminum foil and cooked at 160ºC for 30 min on the top tray in a conventional electric oven (General Electric Deluxe Grill) preheated at 260ºC for 10 min. Cooking proceeded for 10 min after the temperature inside samples reached 75ºC, measured using a thermometer with a probe and an alarm (Incoterm ®). Samples were removed from the oven and the excess humidity on the surface was wiped off using tissue paper. Samples were then cut into 25-mm-thick slices. Eight readings were carried out in each. Instrumental color was analyzed by reflectance in a spectrometer (ColorQuest XE) using D65 as light source, a 10º observation angle in the CIELab system (1978). Results were expressed as angular coordinates L* = luminosity (0 = black; 100 = white), a* (-80 to 0 = green; 0 to + 100 = red) and b* (-100 to 0 = blue; 0 to +70 = yellow). Instrumental measurements of texture were carried out based on the texture profile analysis (TPA), where samples were assessed in a TAXT2 texture analyzer. Samples of each formulation were sliced as described and submitted to the compression test using a 25-kg load. After, samples were pressed to 40% of original thickness using a 50-mm cylindrical probe set at 2.0 mm/s as pretest, 1.5 mm/s as test and 2.0 mm/s return speeds. The texture parameters evaluated were hardness

(strength required to press the sample), cohesiveness (measure of sample deformation before break), chewiness (strength required to chew the sample before swallowing) as described by Tobin et al. (8), and gomosity (energy required to break the semisolid food ready to be swallowed). All analyses were carried out in triplicate. The data obtained were submitted to Analysis of Variance (ANOVA) and the Student-Newman-Keuls (SNK) test in the software SAS version 9.3 (9). III. RESULTS AND DISCUSSION The results of the instrumental color analyses are shown in Table 3. Table 3. Results of instrumental color analyses of fresh

sheep meat sausages Formulation L* a* b*

C 44.31c ±

1.40 9.06a ±

0.93 13.55a ±

0.92

F1 49.92a ±

1.55 6.54c ±

0.66 14.95a ±

1.27

F2 50.66a ±

1.60 6.81c ±

0.51 14.75a ±

0.64

F3 50.70a ±

1.05 6.75c ±

0.19 14.13a ±

0.78

F4 48.87ab ±

2.10 7.44bc ±

1.03 13.95a ±

1.12

F5 46.77bc ±

3.60 7.09bc ±

1.35 15.02a ±

0.87

F6 45.20c ±

2.32 8.04b ±

1.08 14.33a ±

1.19 a b c Means in the same column followed by different lowercase letters differed in the SNK test (p < 0.05).

Except for b*, all color parameters presented variations in formulation means. No statistically significant difference was observed (p > 0.05) in L* values (luminosity) between formulations F1, F2, F3 and F4, and between formulations F4 and F5 and formulations C, F5 and F6. Yalinkiliç et al. (10) analyzed nine sucuk samples (a dried, fermented sausage typically consumed in Turkey) presenting different levels of fat and orange fiber, and noted increased L* values, compared with samples containing orange fiber. Fernandez-Lopez et al. (11) also observed the same results in dried, cured sausages marketed in Spain. The mean a* values (red) in formulation C were different from the others (p < 0.05). No statistically significant difference was observed between formulations F4, F5 and F6, and between formulations F1, F2, F3, F4 and F5 (p > 0.05). In their study, Yalinkiliç et al. (10) observed that mean a* values of samples remained constant, also reported by Fernandez-Lopez et al. (11), which

240

differed from the findings obtained in the present study. Mean b* values (yellow) did not differ (p < 0.05) between formulations. However, the studies cited above reported an increase in b* in samples containing higher orange fiber levels, contrasting with the present study. Instrumental texture results are shown in Table 4. Except for cohesiveness, all texture means varied between formulations. No statistically significant differences (p > 0.05) were seen in hardness between formulations F2, F4, F5 and F6, and between formulations F1, F3, F4, F5 and F6 and the control. Mean chewiness values varied across formulations. No significant difference (p > 0.05) was observed between formulations F1, F2, F4, F5 and F6 and between formulations F1, F3, F4 and F6 and F1, F3 and the control. F2, which was produced with 59% less fat and 50% less salt, presented the highest consistency (hardness and chewiness). According to Bartolomeu (12), tilapia mortadella samples with higher wheat flour contents presented higher hardness.

Table 4. Results of instrumental texture of fresh sheep meat sausage samples

F H C Ch

C 2007.02c ±

216.51 0.58a ± 0.08

998.63c ± 148.22

F1 2849.35b ±

198.96 0.55a ± 0.05

1234.07abc ± 172.80

F2 3475.00a ±

262.42 0.55a ± 0.03

1520.61a ± 52.80

F3 2703.43b ±

373.84 0.49a ± 0.03

1075.60bc ±169.46

F4 3029.40ab ±

256.35 0.53a ± 0.04

1334.25ab ± 215.37

F5 3266.54ab ±

455.07 0.53a ± 0.01

1430.40a ± 221.07

F6 3019.54ab ±

166.24 0.52a ± 0.01

1310.74ab ± 81.81

a b c Means in the same column followed by different lowercase letters differed in the SNK test (p < 0.05). F: Formulations; H: Hardness; C Cohesiveness; Ch: Chewiness.

This finding was also reported by Cardoso (13), in a study that found that hardness is associated with increased fiber levels. This was confirmed in the present study, with samples in which fat was partially replaced with passion fruit meal. IV. CONCLUSION

The formulations prepared with lower fat levels using passion fruit meal, especially F2, presented lower a* values, that is, a pale red product, together with increased b* values (more easily observable yellow tones). F2 was also more resistant, possibly to the gelling, stabilizing and thickening properties of the pectin present in passion fruit meal. ACKNOWLEDGEMENTS The authors thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the grants given, Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) for financial support, and the City Administration of Macaé for the support to carry out this study. REFERENCES 1. He, F. J., Markandu, N. D., Mac Gregor, A. (2005).

Modest Salt Reduction Lowers Blood Pressure in Isolated Systolic Hypertension and Combined Hypertension. Hypertension 46: 66-70.

2. Pompeu, F. R. Tratamento não-farmacológico da hipertensão arterial. Available at: <http://www.medicina.ufmg.br/edump/clm/imphipert.htm>. Accessed on 14/01/2014.

3. Bernardi, D. M., Roman, J. A. (2011). Caracterização sensorial de linguiça Toscana com baixo teor de sódio e análise do consumo de carne suína e derivados na região Oeste do Paraná. Boletim CEPPA 29: 33-42.

4. Junior, J.A.M ; Henry, F. C. ; Valle, F. R. A. F. ; Martins, M. L. L. ; Quirino, C. R. ; Costa, R. S. (2013). Reducing fat and sodium content in pork sausage. African Journal of Biotechnology 12: 3847-3853.

5. Galvan, A. P., Rosa, G., Back, J., Lima, D. P., Corso, M. P. (2011). Desenvolvimento de linguiça tipo Toscana com teor reduzido de gordura e adição de pectina e inulina. In Encontro Paranaense de Engenharia de Alimentos, 3, 2011. Guarapuava/PR.

6. SBC - Sociedade Brasileira de Cardiologia, SBH - Sociedade Brasileira de Hipertensão, SBN - Sociedade Brasileira de Nefrologia. (2007). V Diretrizes Brasileiras de Hipertensão Arterial. Arquivos Brasileiros de Cardiologia, Rio de Janeiro, 89: 24-79.

7. Oliveira, E. M. S. (2009). Caracterização de rendimento das sementes e do albedo do maracujá para aproveitamento industrial e obtenção da farinha da casca e pectina. 2009. Dissertion. (MSc Plant Production). Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes.

8. Tobin, B. D.; O’Sullivan, M. G.; Hamill, R. M.; Kerry, J. P. (2012). Effect of varying salt and fat levels on the sensory and physiochemical quality of frankfurters. Meat Science, Oxford 92: 659-666.

241

http://www.medicina.ufmg.br/edump/clm/imphipert.htm

http://www.medicina.ufmg.br/edump/clm/imphipert.htm

9. SAS. (2009). User’s Guide Statistics. Cary: INSTITUTE SAS.

10. Yalinhiliç, B.; Kaban, G.; Kaya, M. (2012). The effects of different levels of orange fiber and fat on microbiological, physical, chemical and sensorial properties of sucuk. Food Microbiology 29: 255-259.

11. Fernadez-Lopez, J., Viuda-Mortas, M., Sendra, E., Sayas-Barbera, E., Navaroo, G., Perez-Alvares, J. A. (2007). Orange fiber as potential functional ingredient for dry cured sausages. European Food Research and Technology 226: 1-6.

12. Bartolomeu, D. A. F. S. (2011). Desenvolvimento e avaliação da aceitação de embutido defumado “tipo mortadela” elaborado com CMS de tilápia do Nilo (Oreochromis niloticus) e fibra de trigo. 2011, 121 p. Dissertation (MSc, Food Technologies), Federal University Federal of Paraná, Curitiba.

13. Cardoso, C., Mendes, R., Nunes, M. L. (2008). Development of a healthy low-fat fish sausage containing dietary fibre. International Journal of Food Science and Technology 43: 276–283.

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60th International Congress of Meat Science and Technology, 17-22nd August 2014, Punta Del Este, Uruguay

EFFECT OF HIGH PRESSURE PROCESSING ON THE EATING

QUALITY OF LOW FAT SAUSAGE EMULSIONS

Huijuan Yang, Minyi Han, Yun Bai, Xinglian Xu and Guanghong Zhou* Synergetic Innovative Center of Food Safety and Nutrition, College of Food Science and Technology, Nanjing

Agricultural University, Nanjing 210095, PR China

*[email protected]

Abstract – This study investigated the role of

high pressure processing (HPP) for enhancing

the textural properties of sausages with low

fat and no fat replacement. Sausage emulsions

were prepared with levels of fat contents from

zero to 30%. Experiments were carried out to

investigate the effect of 200 MPa HPP on

cooking loss, shear force and the state of

water by low field nuclear magnetic

resonance (LF-NMR). Cooking loss were

significantly reduced (P < 0.05) by HPP.

Textural tests showed that sausages were

exhibited better tenderness at all levels of fat

content treated at 200 MPa than control, and

shear force of sausages treated at 200 MPa

with 20% fat content were similar to the 30%

high fat content. LF-NMR suggested that

HPP significantly changed (P < 0.05) the P2

peak ratio of the three water component.

These findings show that HPP has potential in

transforming the free water (P22) to

immobilized water (P21), resulting in reduced

cooking loss and improved textural properties

of emulsion-type sausages with low fat

content.

. INTRODUCTION

High pressure processing (HPP), is a

relatively new non-thermal food treatment

that can have beneficial effects on the shelf

life and quality of many products (1). With

consumers’ growing awareness of a link between diet and health, HPP used in meat

products with low fat content achieved its

aim mainly by stabilizing substantial of

water and fats in a gel-like matrix, resulting

in some properties similar to those observed

at higher fat levels (2). As 200 MPa HPP

appears to be optimal for many functionality

attributes, this pressure was selected for

determining the optimal minimum fat

content in the range 0 to 30% (3). The main

objectives of this study were to investigate

the effects of HPP on eating properties of

sausage formulations with low fat content.

. MATERIALS AND METHODS

Pork meat from Su Shi group was trimmed

free of connective tissues and excess fat.

The preparation procedures for pork sausage

batters were as followings: added fat levels

were zero, 10, 15, 20 and 30%, protein

levels were maintained constant throughout

by replacing fat with water. 200 MPa

high-pressure processing was carried out in

a 0.3 L capacity 850 Mini FoodLab high

pressure vessel for 2 min at 10°C. In this

work, the raw sausages (with or without

HPP) was termed ‘batter’ and the cooked batters were termed ‘sausages’. NMR was

tested with meat batters, cooking loss and

shear force were tested with sausages.

The data were analyzed using one-way

analysis of variance (ANOVA), and means

were compared by Duncan’s multiple-range

test at 5% level.

. RESULTS AND DISCUSSION

The results of cooking loss of the samples

treated with and without HPP with

increasing fat contents are presented in Fig.

243


1. The lowest cooking losses of the samples

treated with 200 MPa were at 20 and 30%

added fat where losses were just 4 to 5%.

These results suggested that the application

of 200 MPa pressure with samples

containing 15-20% could have beneficial

effects for reducing cooking loss. In

conclusion, the small but significant (P <

0.05) improvements in yields of emulsion

sausages with lower fat contents found here

with the application of HPP is an important

outcome of this study. In this work,

treatment of the samples at 200 MPa is

likely to dissociate of oligomeric structures

into subunits states and partially unfolded

and denatured monomeric structures, as

reported by Bajovic et al. (4). A similar

effect has been reported for HPP (200 MPa)

of low-fat (4-7%) beef sausage batters

where cooking losses were less than 10%

compared with 30% for control (5).

Recently, a similar beneficial effect of HPP

on reducing cooking loss has been reported

for intact beef steak (6).

Fat (%)

0 10 15 20 30

Coo

king

loss

(%

)

0

4

8

12

16

20

0.1MPa200MPa

ab

cd

e

g

f

g

hh

Fig. 1. Changes in cooking losses of pork

sausage containing various fat levels in samples

subjected to 0.1 or 200 MPa pressure for 2 min

at 10°C.

Shear force significantly decreased (P <

0.05) with increasing fat levels both in 200

MPa treated samples and control samples in

Fig. 2. Further, there was no significant

difference (P > 0.05) between the 20% fat

content samples and 30% fat content

samples subjected to 200 MPa for 2 min at

10°C. The reduction in shear force was

probably due to partially protein structure

modification, with unfolding and

subsequent folding. Various researches

investigated the effect of high pressure on

textural properties, and the relationship

between water binding and texture

properties, especially at 200 MPa, results

showed that proper high pressure processing

caused depolymerization, solubilization,

denaturation and aggregation in myofibrillar

(7-8).

Fat (%)

0 10 15 20 30

She

ar fo

rce

(N)

80

100

120

140

160

1800.1MPa200MPa

a

b b

bd

e

d

f

c

f

Fig. 2. Changes in shear force of pork sausage

having various fat levels when subjected to

pressures of 0.1 and 200 MPa for 2 min at 10 °C

before cooking.

The LF-NMR T2 relaxation curves for batter

samples exhibited a multi-exponential

distribution with four distinct water

populations in Tab. 1. T2b1 and T2b2 were

protein-associated water, T21 was

immobilized water, T22 was free water

(9-11). T21, as the relaxation time of the

predominant water component, decreased its

relaxation time from 90.82 to 75.14, and

from 64.28 to 53.87, respectively at control

and 200 MPa treated samples with

increasing fat levels.

244


Tab.1. LF-NMR parameters of meat batters with

low fat after treatment at 0.1 or 200 MPa at 10℃

for 2 min.

Fat T2 relaxation times (ms)

% T2b T21 T22

Pressure 0.1 MPa

0 3.19±0.42a 78.61±4.42a 350.79±5.76bc

10 2.57±0.14bc 70.73±4.41b 351.12±0bc

15 2.17±0.34bc 69.20±3.37b 383.92±25.95a

20 2.16±0.46bc 63.90±2.72c 389.10±20.65a

30 2.01±0.77c 52.58±7.20d 392.02±20.84a

Pressure 200 MPa

0 3.54±0.44a 82.52±1.51a 337.19±17.27c

10 3.27±0.47a 70.67±1.86b 351.48±30.02bc

15 3.21±0.21a 65.75±1.74c 367.25±22.08ab

20 2.39±0.28bc 49.10±4.41d 375.13±24.43ab

30 2.62±0.44b 54.18±3.50d 372.51±17.04ab

Fat Proton compartment sizes (%)

% P2b P21 P22

Pressure 0.1 MPa

0 0.63±0.12g 74.60±3.41f 24.78±3.5a

10 0.78±0.24fg 78.37±2.01e 20.86±2.07b

15 0.94±0.22def 81.35±2.98de 17.73±3.12c

20 1.15±0.21abcd 83.80±1.35bcd 14.99±1.42cde

30 1.22±0.16abc 88.58±4.02a 10.29±0.21f

Pressure 200 MPa

0 0.93±0.12ef 78.01±2.27e 21.07±2.35b

10 1.03±0.03cde 83.35±0.55cd 15.63±0.55cd

15 1.28±0.12ab 86.20±0.98abc 12.53±1.03def

20 1.33±0.18a 86.82±3.58ab 11.87±3.73ef

30 1.09±0.07bcde 85.14±1.36bc 13.79±1.38de

Further, the 200 MPa treated samples were

shorter in relaxation time compared with the

control samples. This revealed that HHP

and increasing fat content both resulted in a

fast relaxation rate for T21, indicating that

both 200 MPa HPP and increasing fat

contents could lead to an increase in

relaxation time, resulting in a migration of

water from the loosely bound water (T22) to

a more tightly bound water (T21).Compared

with the control, pressure at 200 MPa for 2

min at 10°C caused a significant increase (P

< 0.05) in all fat levels. Further, addition of

fat from zero to 20% significantly (P < 0.05)

increased water population P2b1, P2b and P21

in samples treated with 200 MPa, but further

additions to 30% fat showed no further

improvement (P > 0.05). These results

suggested that the application of 200 MPa

pressure with samples containing 15-20%

could have beneficial effects for reducing

extra-myofibrillar water to increase bound

water and myofibrillar water, resulting in

altered cooking loss and textural properties.

. CONCLUSION

This study investigated the application of

HPP (200 MPa) at 10 °C for 2 min to

improve textural properties of pork

emulsion sausages with low fat content. The

application of HPP in sausage emulsions

with low fat content exhibited reduced

cooking loss and shear force, as well as

altered water distribution than control

samples at all fat levels, with a large

improvement for the samples having 15%

and 20% added fat. These results indicated

the suitability of HPP for production of

emulsion sausages having low fat content,

with reduced cooking loss and improved

textural properties.

ACKNOWLEDGMENTS

This work was financed by the National Science

and Technology Support Program

(2013BAD28B03) and the Three New

Agricultural Project of Jiangsu Province

(SX(2011)146).

REFERRENCES

1. Aymerich, T., Picouet, P. A., & Monfort, J.

M. (2008). Decontamination technologies

for meat products. Meat Science, 78(1–2):

114-129.

245


2. Ashie, I. N. A. & Lanier, T. C. (1999).

High pressure effects on gelation of surimi

and turkey breast muscle enhanced by

microbial transglutaminase. Journal of

Food Science, 64(4): 704-708.

3. Villamonte, G., Simonin, H., Duranton, F.,

Cheret, R. & Lamballerie, M. D. (2013).

Functionality of pork meat proteins:

Impact of sodium chloride and phosphates

under high-pressure processing. Innovative

Food Science and Emerging Technologies,

18: 15-23.

4. Bajovic, B., Bolumar, T. & Heinz, V.

(2012). Quality considerations with high

pressure processing of fresh and value

added meat products. Meat Science, 92(3):

280–289.

5. Sikes, A. L., Tobin, A. B. & Tume, R. K.

Use of high pressure to reduce cook loss

and improve texture of low-salt beef

sausage batters (2009). Innovative Food

Science & Emerging Technologies, 10(4):

405-412.

6. Sikes, A. L. & Tume, R. K. (2014). Effect

of processing temperature on tenderness,

colour and yield of beef steaks subjected to

high-hydrostatic pressure. Meat Science, In

press.

7. Bajovic, B., Bolumar, T. & Heinz, V.

(2012). Quality considerations with high

pressure processing of fresh and value

added meat products. Meat Science 92:

280–289.

8. Bak, K. H., Thulstrup, P. W. & Orlien, V.

(2014). Spectroscopic studies on the effect

of high pressure treatment on the soluble

protein fraction of porcine longissimus

dorsi. Food Chemistry 148: 120-123.

9. Bertram, H. C., Whittaker, A.K., Shorthose,

W. R., Andersen, H. J. & Karlsson, A. H.

(2004). Water characteristics in cooked

beef as influenced by ageing and

high-pressure treatment—an NMR micro

imaging study. Meat Science 66: 301–306.

10. Pearce, K. L., Rosenvold, K., Andersen, H.

J., & Hopkins, D. L. (2011). Water

distribution and mobility in meat during

the conversion of muscle to meat and aging

and the impacts on fresh meat quality

attributes-A review. Meat Science, 89(2):

111-124.

11. Sorland, G. H., Larsen, P. M., Lundby, F.,

Rudi, A. P., & Guiheneuf, T. (2004).

Determination of total fat and moisture

content in meat using low field NMR.

Meat Science, 66(3): 543-550.

246


ENHANCED REDDENING OF MEAT BY THE ADDITION OF HIMALAYAN ROCK SALT

Miyuki Kaneko1*, Yasushi Okuda2, Masahiro Waga3,

Toshio Oshida1 and RyoichiSakata1 1School of Veterinary Medicine, Azabu University, Kanagawa 252-5201, Japan

2Frieden, Co., Ltd., Hiratsuka, Kanagawa 252-1201, Japan

3 Itoham Foods Inc., Moriya, Ibaraki302-0104, Japan

*[email protected]

Abstract – This study was carried out to

examine the reddening of meat products

through the addition of natural yellow salt,

(YS). Following YS or NaCl addition at

2% to pork subsequent to nitrite (0~100

ppm) treatment, color was analyzed both

visually and spectrophotometrically.

Assessment was also made of heme

pigment content in meat. YS was found to

bring about greater reddening compared

to NaCl, thus indicating residual nitrite

and nitrate content to be significantly

higher, through the amount of either was

quite small. The color forming ratio in

meat was found to change by heme

pigment content and the amount of nitrite

required for red coloring was noted to

vary significantly.

I. INTRODUCTION

Attention is increasingly being directed to

food safety in response to consumer demand

for this regard and accordingly, techniques

for effectively enhancing red coloring using

coloring agents as little as possible have thus

become essential (1,2). Natural salts,

particularly such as those found in

Himalayan salts appear to be promising

candidates for reddening owing to their

mineral content. To effectively bring about

this coloring in meat products, examination

was made of Himalayan rock salt for

comparison with ordinary cooking salts.

II. MATERIALS AND METHODS

Meat taken from pig thigh was depleted as

much as possible of its fat and connective

tissue and then minced. To the meat samples,

YS or NaCl was added at 2% along with

0.1% sodium ascorbate and sodium nitrite in

the concentration range, 0, 10, 30, 50 or 100

ppm. The samples were then cooked at 75ºC

for 30 min after for 4 days storage at 4ºC

under anaerobic conditions. After sample

cooling, color was assessed visually and then

with a spectral colorimeter. The color

forming ratio (CFR) (3) and heme pigment

content were measured by acetone extraction.

Residual nitrite and nitrate content were

determined according to Mirna and Schütz

(4) and by copper-cadmium column

reduction, respectively.

III. RESULTS AND DISCUSSION

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http://ejje.weblio.jp/content/veterinary+medicine


In the case of NaCl addition without nitrite,

CFR was found to be less than that in YS

samples and a* (redness) was also less than

that in other samples. In YS samples with and

without nitrite, meat color was red and CFR

was found to exceed 70% regardless of the

low addition of nitrite in the range, 0 to 100

ppm (Fig.1).

Fig.1. Color forming ratio (CFR).

Compared to NaCl samples, residual nitrite

content tended to be higher in the case of YS

addition (Fig.2), as also noted for nitrate

included essentially in the rock salt. YS is

thus shown to effectively enhance meat

reddening. Small amounts of nitrite, nitrate

and minerals may possible have contributed

to this finding.

Fig. 2. Residual nitrite levels in samples.

CFR was seen to change with increase in

heme pigment content in pork, in the case of

YS addition. When the amount of this

pigment was quite small, the color ratio

increased in the samples (Fig.3). The limited

capacity of heme nitrosation for enhancing

meat reddening may possibly be the reason

for this.

Fig.3. CFR as a function of heme pigment content.

IV. CONCLUSION Using Himalayan rock salt, enhanced meat

reddening was achieved at only very small

nitrite content or even in its absence. The

color forming ratio in cooked meat changed

with heme pigment content in meat owing to

the low degree of nitosation in the meat.

ACKNOWLEDGEMENTS The authors are thankful to Azabu University,

Japan, for kindly awarding a research project

grant.

REFERENCES 1. Mancini, R.A. & Hunt M.C (2005). Current

research in meat color. Meat Science

71:100-121.

2. Sakata, R. (2010). Prospects for new

technology of meat processing in Japan. Meat

Science 86: 243-248.

3. Sakata, R. Ohso, M & Nagata, Y. (1981). Effect

of porcine muscle conditions on the color of

0

10

20

30

40

50

60

70

80

90

0 50 100

CF

R(%

)

Added nitrite (ppm)

NaCl samples

YS samples

0

10

20

30

40

0 50 100

Resi

du

al

nit

rite

(pp

m)

Amount of added nitrite (ppm)

YS samples

NaCl samples

50

55

60

65

70

75

80

0 10 20 30

CF

R(%

)

Amount of added nitrite (ppm)

Low levels of heme

High levels of heme

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cooked cured meat products. Agricultural

Biological Chemistry 45: 2077-2081.

4. Mirna A. & Schütz G. (1972). Verfahren zur

gleichzeitigen Bestimmung des Pökelfarb-

stoffes sowie von Nitrit und Nitrat in

Fleischerzeugnissen. Fleischwirtschaft 52:

1337-1338.

249


EFFECT OF MEAT AND NON-MEAT PROTEIN MIXING RATIO ON QUALITY CHARACTERISTIC OF EMULSION-TYPE SAUSAGE

Geunho Kang*, Hyoung-Joo Ham, Soohyun Cho, Pil-Nam Seong, Sun-Mun Kang, Kyoungmi

Park, Hoa Van Ba and Beom-Young Park

Animal Products Research and Development Division, National Institute of Animal Science, RDA, Suwon, Republic of Korea. *Corresponding author (phone: +82-31-290-1684; fax: +82-31-290-1697; e-mail: [email protected])

Abstract – This study was conducted to assess the effect of meat/non-meat protein mixing ratio the on quality characteristic of emulsion-type sausage. The sausages were formulated with various mixing ratios of meat and non-meat ingredients. The results showed that hardness was significantly (p<0.05) lower in sausage with pork (C) and sausage with more soybean (T4) in comparison to the other treatments. Springiness was significantly higher in sausage with more corn starch (T3) but lower in T4 sample in comparison to the other treatments. The TBARS values were significantly (p<0.05) lower in the C and sausage with chicken (T2) in comparison to the other treatments throughout the cold storage time. Also, the T4 samples showed significantly (p<0.05) lower TBARS value than other samples until 4 weeks of cold storage. These results indicate that the texture and lipid oxidation of emulsion-type sausage were affected by meat/non-meat protein ratio. Key Words – sausage, meat, non-meat protein I. INTRODUCTION There has been an increase interested in healthier processed food products by consumers. Therefore, reduction of saturated fat content has been an emphasis since it is not beneficial to human health [1-2]. Recently, research trends in meat products relates to replacing pork fat or reducing sodium content [3-5]. In Korea, there are several different types of sausages such as non-emulsion type, called ‘press ham’ or ‘mixed press ham’, and emulsion-type sausage. The materials used for the emulsion-type sausage manufacture are often pork, chicken, fish etc, and non-meat contents such as starch, corn, soybean etc. Depending on the meat/ non-meat materials mixing ratio the quality and price of final products are also different. Kang et al. [6] recently reported that moisture content and texture attributes of sausage were affected by

pork/duck meat mixing ratio. However, it is still unknown whether the meat/non-meat ingredients mixing ratio affects the quality characteristics of emulsion-type sausage. Therefore, the objective of this study was to investigate the effect of meat/non-meat protein mixing ratio on quality characteristic in emulsion type sausage. II. MATERIALS AND METHODS Treatments of sausages were subjected to 82% pork meat + 18% additives (C), 72% pork meat + 10% corn starch + 18% additives (T1), 72% pork meat + 10% chicken meat + 18% additives (T2), 62% pork meat + 10% corn starch + 10% chicken meat (T3), 80% pork meat + 2% concentrated soybean protein + 18% additives (T4), and 60% pork meat + 10% corn starch + 10% chicken meat + 2% concentrated soybean protein + 18% additives (T5). Three batches of emulsion-type sausages (20 kg each) were manufactured for each treatment. The meat was prepared with a meat grinder. All materials were treated with mixing, curing/ripening, emulsion, stuffing, and cooking. After cooking, the sausage samples were used for the chemical composition, texture and quality characteristics analyses. Chemical composition was performed according to the method of Anderson et al. [7]. Texture of sausages was analyzed by texture instrument (5543, Instron, USA). TBARS of all samples determined according to the method of Buege and Aust [8]. III. RESULTS AND DISCUSSION Table 1 shows the effect of different meat/non-meat protein mixing ratios on texture of emulsion-type sausage. Hardness was significantly (p<0.05) lower in C and T4 samples compared to the other treatments, whereas no differences in cohesiveness

250


among the treatments were observed. Springiness was significantly higher in T3 but significantly lower in T4 compared to the other treatments. These results indicate that hardness and springiness of emulsion-type sausage were affected by the mixing ratio. Table 1 Formulation (%) for manufacture of sausage used in the experiment Ingredients C T1 T2 T3 T4 T5

Pork meat 82 72 72 62 80 60

Corn starch 0 10 0 10 0 10

Chicken meat 0 0 10 10 0 10

Concentrated soybean protein

0 0 0 0 2 2

Pork fat 15 15 15 15 15 15

Phosphate 0.3 0.3 0.3 0.3 0.3 0.3

Salt 1.25 1.25 1.25 1.25 1.25 1.25

Sugar 0.75 0.75 0.75 0.75 0.75 0.75

Black pepper 0.2 0.2 0.2 0.2 0.2 0.2

L-ascorbic acid 0.05 0.05 0.05 0.05 0.05 0.05

Garlic powder 0.2 0.2 0.2 0.2 0.2 0.2

L-Glutamate 0.25 0.25 0.25 0.25 0.25 0.25

Total 100 100 100 100 100 100

Table 2 Effect of different meat and non-meat protein

mixing ratios on texture of emulsion-type sausage

Treatments* Hardness

(kg) Cohesiveness

(%) Springiness

(mm)

C 0.47D 1.20 14.21AB

T1 0.63B 1.32 14.15AB

T2 0.54C 1.19 14.69AB

T3 0.67A 1.18 15.12A

T4 0.47D 1.30 13.83B

T5 0.65AB 1.47 14.69AB

SEM 0.01 0.07 0.15 A-DMeans with different letters within the same column differ (p<0.05). Table 2 shows changes in TBARS value of emulsion-type sausage manufactured with different mixing ratio of meat and non-meat protein ingredients during cold storage. The TBARS value was significantly (p<0.05) lower in C and T2 samples compared to the other samples throughout the cold storage time. Also, T4 sample

showed significantly (p<0.05) lower TBARS value than other samples until 4 weeks of cold storage. Therefore, these results suggest that the TBARS value of emulsion-type sausage was affected by the meat/non-meat protein types mixing ratio. Table 3 Changes in TBARS of emulsion-type sausage

manufactured with different meat/non-meat protein mixing ratio during cold storage

Treat-ments*

Storage time (weeks)

1 2 3 4 5

C 0.92Cc 1.1Bb 1.27Ca 1.14Cb 0.65Cd

T1 2.43Bb 3.02Aa 2.77Bab 2.85Bab 3.1Aa

T2 0.81Cb 0.88Bab 0.99Ca 1.02Ca 0.76BCb

T3 3.07AB 3.08A 3.11AB 3AB 2.99A

T4 1.52C 1.2B 1.12C 1.09C 1.01B

T5 3.55Aa 3.3Aab 3.31Aab 3.35Aab 3.02Ab

SEM 0.22 0.2 0.19 0.19 0.21 A-CMeans with different letters within the same column differ (p<0.05). a-dMeans with different letters within the same row differ (p<0.05). IV. CONCLUSION Our study results indicate that texture (hardness and springiness) and lipid oxidation (TBARS) of emulsion-type sausage were considerably affected by the meat/non-meat protein types mixing ratio. Our study results indicate that sausages with meat protein were lower lipid oxidation compared to the sausages with non-meat protein during cold storage. Therefore, this data suggest that sausage manufactured with only meat protein is well beneficial to the health of consumers. ACKNOWLEDGEMENTS This research was supported by a grant from the Korean Rural Development Administration. REFERENCES 1. Food Standards Agency (FSA) (2009). Draft

recommendations on promotion of lower fat products including dairy products, fat and saturated fat reductions for meat products, and portion size availability for savoury snacks. UK: Food Standards Agency.

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2. Ospina-E, J. C., Sierra-C, A., Ochoa, O., Perez-Alvarez, J. A. & Fernandez-Lopez. J. (2012). Substitution f saturated fat in processed meat products: A review. Critical Reviews in Food Science and Nutrition, 52: 113-122.

3. Asuming-Bediako, N., Jaspal, M. H., Hallett, K., Bayntun, J., Baker, A. & Sheard, P. R. (2014). Effects of replacing pork backfat with emulsified vegetable oil on fatty acid composition and quality of UK-style sausages. Meat Science 96: 187-194.

4. Marchetti, L., Andés, S. C. & Califano, A. N. (2014). Low-fat meat sausages with fish oil: Optimization of milk proteins and carrageenan contents using response surface methodology. Meat Science 96: 1297-1303.

5. Mora-Gallego, H., Serra, X., Guàrdia, M. D. & Arnau, J. (2014). Effect of reducing and replacing pork fat on the physicochemical, instrumental and sensory characteristics throughout storage time of small caliber non-acid fermented sausages with reduced sodium content. Meat Science 97: 62-68.

6. 6. Kang, G., Cho, S., Seong, P., Kang, S., Park, K., Park, B. & Kim, J. (2013). Effect of mixture ratio of pork and duck meat on quality characteristic in comminuted sausage. In proceedings 59th International Congress of Meat Science and Technology (p. 138), 18-23 August 2013, Izmir, Turkey.

7. Anderson, S., Aldana, S., Beggs, M., Birkey, J., Conquest, A., Conway, R., Hemminger, T., Herrick, J., Hurley, C., Ionita, C., Longbind, J., McMaignal, S., Milu, A., Mitchell, T., Nanke, K., Perez, A., Phelps, M., Reitz, J., Salazar, A., Shinkle, T., Strampe, M., Van Horn, K., Williams, J., Wipperfurth, C., Zelten, S. & Zerr, S. (2007). Determination of fat, moisture, and protein in meat and meat products by using the FOSS FoodScan™ Near-Infrared Spectrophotometer with FOSS artificial neural network calibration model and associated database: collaborative study. Journal of AOAC International 90: 1073-1082.

8. Buege, J. A. & Aust, J. D. (1978). Microsomal lipid peroxidation. Methods in Enzymology 52:302-307.

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60th International Congress of Meat Science and Technology, 17-22rd August 2014, Punta Del Este, Uruguay THE POTENTIAL ROLE OF ARGININE IN COMPETITIVENESS AND FUNCTIONALITY OF COAGULASE-NEGATIVE STAPHYLOCOCCI DURING MEAT FERMENTATION

Frédéric Leroy1*, Maria Sanchez-Mainar1, Stefan Weckx1 & Luc De Vuyst1

1 Research Group of Industrial Microbiology and Food Biotechnology (IMDO), Faculty of Sciences and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium

* [email protected] The capacity for arginine metabolism during meat fermentation offers a potential bonus in fitness to coagulase-negative staphylococci (CNS) in the absence of carbohydrates, as is particularly so for the arginine deiminase (ADI) pathway. An alternative is provided by the nitric oxide synthase (NOS) enzyme, which converts arginine into citrulline and nitrogen oxide (NO). Since NO causes colour-stabilizing nitrosylation of myoglobine, this pathway could potentially be used for the generation of nitrosomyoglobine in clean label products without added nitrate and/or nitrite. A genotypic and physiological screening of 88 CNS strains indicated that active ADI machinery is often found in this CNS group, although in a strain-specific manner. In contrast, phenotypic NOS-like activity was only present in one of the strains (Staphylococcus haemolyticus G110), although the genetic potential for NOS was widespread among CNS strains. Attempts to express NOS activity in some of the latter strains were unsuccessful, suggesting that the genetic potential for NOS is not commonly expressed by CNS. Both ADI and NOS kinetics were investigated in a meat simulation medium, indicating the need for sufficient oxygen for NOS activity. The use of NOS-positive CNS cultures as a curing alternative in fermented meats is thus not clear-cut. Key Words – meat fermentation, coagulase-negative staphylococci, arginine deiminase, nitric oxide synthase, colour I. INTRODUCTION During the production of traditionally fermented sausages, spontaneous fermentation takes place that is dominated by communities of lactic acid bacteria (LAB) and catalase-positive cocci, mostly coagulase-negative staphylococci (CNS) (1). During this process, LAB cause a desirable acidification of the sausage batter, whereas CNS produce a stable cured colour via nitrate reductase activity,

generate aroma, and protect against oxidation. In such spontaneous processes, Lactobacillus sakei is the most dominant LAB species and thus frequently used as a starter culture for meat fermentation. A wider species variety exists amongst the meat-associated CNS communities but, in general, Staphylococcus xylosus, S. equorum, and S. saprophyticus are predominant. Yet, several other CNS may also occur (1). Because meat is generally poor in carbohydrates and added sugars are rapidly depleted, differences in the use of alternative energy sources may lead to improved competitiveness for certain meat bacteria (2). The arginine deiminase (ADI) pathway, which is present in certain CNS, is an interesting option, as it generates ATP and protects against acid stress through ammonia production. Whereas ornithine is the end-metabolite, the intermediate citrulline may be partly excreted too, as is the case for Lb. sakei (3). An alternative but poorly explored arginine-converting pathway could potentially be based on the action of nitric oxide synthase (NOS) (4). As a result of NOS activity, arginine is converted into citrulline and nitric oxide (NO). From a technological point of view, this could be of interest for colour formation in fermented meat products prepared without nitrate and nitrite (4). Indeed, the cured colour of fermented meat products is obtained via the formation of nitrosomyoglobin, resulting from the interaction between muscle-based myoglobin and NO that generally originates from the addition of nitrate and/or nitrite as curing agents under low pH conditions. Thus far, most of the studies looking into colour generation via NOS in fermented meat

253

products have been rather preliminary and definite proof is lacking (4, 5). Therefore, the present study investigated the use of arginine via ADI and NOS activities by CNS and explored the heterogeneity on species and strain level, both from a phenotypical and genotypical perspective. It also explored the kinetic patterns of arginine metabolism as a function of the prevailing glucose and oxygen levels. II. MATERIALS AND METHODS A genotypic screening for nos (NOS-encoding) and arcA (ADI-encoding) genes was performed in a set of 88 strains of CNS. Four primer sets for the nos gene and one primer set for the arcA gene were designed, based on Genbank entries for staphylococci, and the associated PCR assay conditions were optimised. Second, a phenotypic screening of the ability of these CNS strains to use the precursor arginine was carried out. Each CNS strain was cultivated at 30 °C in 100 ml of meat simulation medium (MSM), containing 3 g/l of added arginine (17.2 mM), under microaerobic or aerobic (shaker incubator) conditions, with sampling after 72 h. Third, a further screening under possibly NOS-stimulating conditions [including growth on solid agar, the use of different temperatures (12, 20, and 40 ºC) and initial pH values (pH 6.5, 7.0, and 7.5), and the presence of added glucose, hemin, methanol, myoglobin, different minerals (Fe++, Mg++, Ca++), and tetrahydrobiopterin] was performed on a subset of 12 nos-positive S. carnosus strains. Fourth, kinetic analyses of possible NOS activities in CNS (S. haemolyticus G110, S. pasteuri αS3-13, and S. carnosus 833), as a function of glucose and oxygen levels, were done in laboratory fermentors. Fermentations were carried out at 30 °C in 10 l of MSM containing 3 g/l of added arginine and 5 g/l of added glucose, and with an initial pH of 5.8 under micro-aerobic or aerobic conditions. Bacterial growth was followed by the determination of colony forming units (cfu) per ml on mannitol salt agar (MSA).

In all experiments, concentrations of arginine, citrulline, and ornithine were measured with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Glucose consumption was measured with high-performance liquid chromatography (HPLC) with refractive index (RI) detection. III. RESULTS AND DISCUSSION Among the 88 strains of CNS tested, the nos gene was present in 74 % of the strains. The genetic potential for ADI activity was also frequently found, i.e., amongst 44 % of the CNS strains tested. The phenotypic screening experiments confirmed that arginine metabolism was indeed common, resulting in mixtures of citrulline and mostly ornithine, even though considerable variability was found on species and even strain level. Although microaerobic conditions seemed to stimulate ADI activity, this pathway generally also took place under aerobic conditions. The production of citrulline without ornithine formation, indicative of potential NOS activity, was not found under the conditions tested for any of the CNS strains, except for S. haemolyticus G110. The latter strain showed modest citrulline production (9 ± 4 mM) without ornithine under aerobic conditions. The absence of phenotypic NOS activity amongst the other strains was remarkable, considering the fact that the nos gene was quite often present. Further attempts to express the nos potential, by applying growth on solid agar or by modifying the medium composition or the prevailing temperature and pH, were unsuccessful. This indicates that the genetic potential for NOS activity is not readily expressed under a range of conditions. Kinetic experiments in MSM confirmed that NOS-like activity, indicated by citrulline production without ornithine, was obtained with S. haemolyticus G110 (Fig. 1). This pattern was found under aerobic conditions, but no manifest consumption of arginine was seen under micro-aerobic conditions, further confirming the NOS-like behaviour of the strains since this pathway requires oxygen. With S. pasteuri αS3-13 and S. carnosus 833, combinations of citrulline and ornithine were obtained under both aerobic and micro-aerobic

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conditions, indicating ADI activity. The example of S. carnosus 833 is given in Fig. 2, but results for S. pasteuri aS3-13 were similar. ADI activity only started after glucose depletion, indicating carbon catabolite repression. This suggests a role for ADI activity in the CNS metabolism of alternative energy substrates under glucose-depleted conditions, commonly encountered in meat fermentation.

Fig. 1. Conversion of arginine (mM; circles) into citrulline (mM; triangles) by S. haemolyticus G110 [cell counts in log (cfu/ml); squares] under aerobic (full symbols) and micro-aerobic conditions (open

symbols) in MSM at 30 °C. The arrow indicates the moment of glucose depletion for both cases.

Fig. 2. Conversion of arginine (mM; circles) into citrulline (mM; triangles) and ornithine (mM;

diamonds) by S. carnosus 833 [cell counts in log (cfu/ml); squares] under aerobic (full symbols) and micro-aerobic conditions (open symbols) in MSM

at 30 °C. The arrow indicates the moment of glucose depletion for both cases.

Although NOS activity leading to NO generation could be of interest for colour formation in fermented meats with clean labels, its successful implementation in fermented meat products has thus far been insufficiently proven (5). It remains unclear to which degree NOS activity can be modulated by environmental conditions and process factors. The need for oxygen represents a major potential hurdle, besides the fact that expression of NOS activity is absent in the large majority of the CNS strains possessing the required gene under a wide range of tested conditions. IV. CONCLUSION Arginine consumption by CNS generally relied on the ADI pathway, although considerable variability existed on species and strain level. The ADI pathway only became active in the absence of glucose, which has physiological implications with respect to ecosystem adaptation. The genetic potential for NOS activity was present in many CNS strains too, but this was not reflected in the phenotypic behaviour, except for one strain, indicating that the bottleneck was likely to be on the gene expression level. The use of NOS-positive CNS cultures for nitrate and nitrite cutback in fermented meats is therefore not straightforward and its industrial application could be problematic. From an ecological perspective, further research is needed to assess to which degree differences in arginine metabolism play a role in the establishment of specific CNS communities during meat fermentation. ACKNOWLEDGEMENTS The authors acknowledge their financial support of the Research Council of the Vrije Universiteit Brussel (OZR, SRP, IRP, and IOF projects, and in particular the HOA project 'Artisan quality of fermented foods: myth, reality, perceptions, and constructions'), the Hercules Foundation (project UABR 09/004), the Research Foundation-Flanders (project FWOAL632), and Flanders' FOOD (project NITRILOW). REFERENCES 1. Ravyts, F., De Vuyst, L. & Leroy, F. (2012).

Bacterial diversity and functionalities in food

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fermentations. Engineering in Life Sciences 12: 356-367.

2. Janssens, M., Van der Mijnsbrugge, A., Sánchez Mainar M., Balzarini T., De Vuyst L. & Leroy, F. (2014). The use of nucleosides and arginine as alternative energy sources by coagulase-negative staphylococci in view of meat fermentation. Food Microbiology 39: 53-60.

3. Rimaux, T., Vrancken, G., Pothakos, V., Maes, D., De Vuyst, L. & Leroy, F. (2011). The kinetics of the arginine deiminase pathway in the meat starter culture Lactobacillus sakei CTC 494 are pH-dependent. Food Microbiology 28: 597-604.

4. Morita, H., Yoshikawa, H., Sakata, R., Nagata, Y. & Tanaka, H. (1997). Synthesis of nitric oxide from the two equivalent guanidino nitrogens of L-arginine by Lactobacillus fermentum. Journal of Bacteriology 179: 7812–7815.

5. Sudhamsu, J., & Crane, B. (2009). Bacterial nitric oxide synthases: what are they good for? Trends in Microbiology 17: 212-218.

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60th International Congress of Meat Science and Technology, 17-22rd August 2014, Punta Del Este, Uruguay EFFECT OF MUSHROOM INCORPORATION ON QUALITY, SENSORY

AND MICROBIAL STABILITY OF FRANKFURTER TYPE-SAUSAGE Pil-Nam Seong, Kyoungmi Park, Soohyun Cho, Geunho Kang, Sun-Mun Kang, Beom-Young

Park and Hoa Van Ba*

Animal Products Research and Development Division, National Institute of Animal Science, RDA, Suwon, Republic of Korea. *Corresponding author (phone: +82-31-290-1684; fax: +82-31-290-1697; e-mail: [email protected])

Abstract –This study was carried out to investigate the effect of Tricholoma matsutake mushroom addition on the quality, sensory and microbial stability of frankfurter during storage (1 and 14 d) at 4oC. Various levels (1, 2.5 and 4.5%) of ground mushroom were incorporated into frankfurters. Frankfurters with no added mushroom were used as control. Our results revealed that lightness was not affected by the mushroom addition but the redness decreased whereas the yellowness increased with increased mushroom levels. The addition of mushroom resulted in improvements of taste and flavor; sausages with higher mushroom level had higher flavor and taste scores (p< 0.05), whereas sausages formulated with 0.01% NaNO2 had the lowest flavor and taste scores. Also, the addition of mushroom reduced the number of aerobic bacteria in the sausages during storage. These results suggest that T. matsutake mushroom can be used as a natural functional ingredient to improve taste and flavor as well as shelf-life stability of frankfurter. Key Words – frankfurter type-sausage, mushroom

I. INTRODUCTION

The currently available chemical additives (e.g., antioxidants and antimicrobials) have been suspected to cause toxicity problems that negatively affect consumer’s health (1). Therefore, a new trend to substitute these synthetic compounds with bioactive compounds from natural sources have been received the most attention by consumers and meat processors (2). In recent times, the consumed amount of mushrooms has considerably increased, involving a large number of species. Of which, Tricholoma matsutake is widely distributed in Asian countries and has been regarded as famous foods due to its pleasant flavor, taste and high nutritional quality such as high levels of proteins, carbohydrates, minerals and vitamins as well as low fat (3, 4). Besides, T. matsutake has been demonstrated to contain significant amounts of compounds such as polysaccharides with strongly bioactive properties (4). Though the T. matsutake has a pleasant flavor and may exert bioactive properties in sausages, which can be effectively incorporated into sausage. However, very little attention has been paid to the use of edible

mushroom as functional ingredients in meat products. Therefore, the objective of this study was to evaluate the effect of addition of T. matsutake on the quality, sensory and microbial stability of frankfurter type-sausage during refrigerated storage.


Mushroom and frankfurter preparation Fresh T. matsutake mushroom purchased from a local supermarket (Suwon, Korea) was washed and then ground in a blender. The refrigerated pork meat and back fat were separately ground through a 3 mm plate. All sausages were prepared with same levels of following ingredients: 50% pork, 28% back-fat, 20% ice water, 0.5% phosphate and 1.5% salt. The following combinations were used; T1: 1% mushroom, T2: 2.5% mushroom, T3: 4.5% mushroom, and T4: 100ppm NaNO2. Ice water in T1, T2 and T3 was replaced by the added mushroom levels. Three batches (each 5 kg) were prepared for each treatment and the batches served as the control (C) were not added mushroom. The meat batters were prepared and then stuffed into 28-mm diameter collagen casings using a vacuum stuffer, and finally were cooked at 80 °C for 40 min in a smokehouse. The cooked sausages were cooled and then sealed in polyethylene/polyamide pouches. The quality and sensory characteristics were investigated on the day after processing (1 d) and the samples stored at 1 and 14 d at 4 °C were used to investigate microbial stability Color, microbial and sensory analysis Color values (CIE L*, a*, b*, chroma and hue) were measured at 3 different locations on the freshly cut surface of each sausage sample using a Minolta Chroma Meter CR-400 (Osaka, Japan). Aerobic bacteria were determined on 3M Petrifilm (HealthCare, MN, USA), and each sample was repeated 3 times. Sensory traits were evaluated by trained panel 6 members using the method of Deda et al. (5). Statistic analysis Data were analyzed using GLM procedure of SAS program (2007).

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60th International Congress of Meat Science and Technology, 17-22rd August 2014, Punta Del Este, Uruguay III. RESULTS AND DISCUSSION

Table 1 Effect of mushroom incorporation on color of frankfurter-type sausages

item

Added level

0 %

1% 2.5% 4.5%

0.01% nitrite

SEM F.

value

L* 72.71 73.27 73.08 73.11 74.47 0.2 5.65*

a* 2.99 2.83 2.69 2.42 4.86 0.06 12.92*

b* 9.41 9.75 10.15 10.77 9.14 0.11 7.22*

C* 9.88 10.16 10.50 11.04 10.36 0.12 3.72*

H 72.36 73.82 75.16 77.37 62.01 0.29 11.90*

SEM: standard error of mean; C*: chroma; H: hue; *, p < 0.05.

Table 1 shows the L*(lightness), a*(redness), b*(yellowness), C* (chroma) and h (hue) values of frankfurters formulated with various fresh mushroom levels and 0.01% NaNO2. No differences among the treatment groups were found for the lightness (p>0.05) but the redness decreased whereas the yellowness increased with increased mushroom levels. However, sausages made with 0.01% NaNO2 showed the highest a* values and lowest b* values. The addition of mushroom resulted in improvements of taste and flavor of frankfurter-type sausages; higher added mushroom level sausages had higher flavor and taste scores (p< 0.05), whereas sausages formulated with 0.01% NaNO2 had the lowest flavor and taste scores (Table 2). Table 2 Effect of mushroom incorporation on sensory characteristics of frankfurter-type sausages

Added level

item 0 %

1% 2.5% 4.5%

0.01% nitrite

SEM F.

value

Flavor 3.78 3.94 4.0 4.28 3.67 0.65 5.5*

Texture 3.39 3.89 3.67 4.33 3.39 0.90 1.65

Taste 3.61 3.72 3.72 3.94 3.28 0.92 8.2*

Acceptability

3.44 3.89 3.61 4.06 3.11 0.40 2.97

SEM: standard error of mean; *, p < 0.05; Rating scale: 7=extremely desirable, 1=extremely undesirable. Furthermore, the incorporation of mushroom significantly reduced the number of aerobic bacteria in sausages during cold storage in comparison to the control however the sausages made with 0.01% NaNO2 showed higher bacteria inhibition capacity (Fig. 1). The improvements of flavor and taste of sausages could be due to umami-taste active components in mushroom (3) while the

antimicrobial activity of the T. matsutake could be due to the presence of essential bioactive compounds as most of them exhibited antimicrobial activity.

Fig. 1. Effect of mushroom incorporation on aerobic

bacteria in frankfurter-type sausages during storage at 4oC.

IV. CONCLUSION

Based on the results obtained in the present study, it is concluded that T. matsutake mushroom incorporation into frankfurter-type sausage formulations resulted in improvement of flavor and taste, and shelf-life stability due to reduction of aerobic bacteria growth.

ACKNOWLEDGEMENTS

This work was supported by "Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ00937401)" Rural Development Administration, Republic of Korea. REFERENCES 1. Sherwin, E. R., Branen, A. L., Davidson, P. M., &

Salminen, S. (1990). Food additives New York: Marcel Dekker.

2. Ahn, J., Grun, I. U., & Mustapha, A. (2004). Antimicrobial and antioxidant activities of natural extracts in vitro and in ground beef. Journal of Food Protection 67: 148–155.

3. Cho, I. H., Choi, H. K., Kim, Y. S. (2010). Comparison of umami-taste active components in the pileus and stripe of T. matsutake-mushrooms of different grades. Food Chemistry 118: 804–807.

4. Tong, H., Liu, X., Tian, D., Sun, X. (2013). Purification, chemical characterization and radical scavenging activities of alkali-extracted polysaccharide fractions isolated from the fruit bodies of Tricholoma matsutake. World Journal of Microbiology and Biotechnology 29:775–780.

5. Deda, M. S., Bloukas, J. G., & Fista, G. A. (2007). Effect of tomato paste and nitrite level on processing and quality characteristics of frankfurters. Meat Science 76: 501–508.

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OPTIMISING THE SHELF-LIFE OF PORK CHOPS WITH NEW GAS COMPOSITIONS AND REDUCED HEADSPACE

Bozec A, Vautier A, L’Hommeau T, Le Roux A. IFIP, meat quality and safety, Le Rheu, France

*[email protected]

Abstract – The objective of this study was to evaluate the influence of several gases compositions and headspaces on pork chops packages. Three batches of fresh pork meat packed under Modified Atmosphere Packaging (MAP) were studied. The evolutions of the gas composition, pH during storage were monitored and the evolutions of the spoilage microflora were evaluated. A panel of consumers realized sensorial analyses. The composition of gases followed the same evolution in the two headspaces. There are no statistical differences between headspace 1 (H1) and H2. The H1 allowed an equivalent conservation H2. Consumers prefer smaller trays (H1). The best purchase intent (78%) is obtained by containing the lowest oxygen content (40%). The growth of Enterobacteriacae and Brochothrix shows respectively a 1 log and 2 log reduction with increasing CO2 rate (60% vs 30%). Small headspace (H1) with both 50%-50% and 60%-40% O2/CO2 mixture induced an overall microbiological quality that is in agreement with the GHP recommendations, and allows a 4 days extension of the shelf life.

INTRODUCTION Nowadays consumers are looking for fresh, tasty and safe to eat meat products, and want them in a convenient, easy-to-choose tray. Because consumers use meat color as an indicator of wholesomeness, recent researches in MAP have been focused on finding the correct gas mixture. Objectives are to maximize initial color, color stability, and shelf life while also minimizing microbial growth, lipid oxidation, and gaseous headspace (Mancini et al, 2005). Jakobsen et al (2000) reported that while oxygen levels higher than 20% were necessary to promote meat color, package oxygen contents higher than 55% did not result in additional color stabilizing benefits. The effectiveness of MAP is generally determined by the amount of available CO2. CO2 dissolves into the aqueous part of the product in traditional MA package resulting in a volume contraction of a flexible package. The package collapse is usually reduced by lowering the CO2 partial pressure by introducing gases with significantly less solubility, that is, N2 or O2, in the MA. This is, however, not

optimal for packaged products (Bjørn Tore Rotabakk, 2007). Suppliers of packaging are actively improving environmental performance through total material utilization, recycling or reuse of industrial scrap. These actions contribute to decrease packaging waste through progressively thinner, lighter, yet stronger materials. Another way to limit environnemental impact is to reduce the height of the tray with the optimization of the headspace. According to (Kenneth, et al 2008), headspace gas must be approximately 1.5–2 times the meat volume and package collapse is generally thought to be prevented by headspace gas to meat volumes of 2 to 3 (Gill & Gill, 2005). The first objective of this study was to investigate the effect of different gas compositions and headspaces (four-gas MAP, two headspaces) on shelf-life and consumers perception of fresh pork packaging. The second objective was to confirm shelf-life with new determined gas compositions and optimized headspace.

MATERIALS AND METHODS Three batches of fresh pork chops were packed in four different gas compositions using two different headspaces, then analyzed during 9 days of storage. For each of the 3 repetitions, pigs were selected according to weight (kg) and carcass composition (TMP). The same slaughter process was used for all the batches: slaughtering on Monday, pH24, cutting, deboning, shell freezing and slicing on Tuesday then MApacking on Wednesday (day 0). Packing: 2 x chops (150 g) MApacked in four gas compositions: 1. 70% O2 + 30% CO2 (70/30); 2. 60% O2 + 40% CO2 (60/40); 3. 50% O2 + 50% CO2 (50/50); 4. 40% O2 + 60% CO2 (40/60) on a tray sealer (Guelt, OPS1000, France). The trays used were, PS EVOH PE 11000 (Form’ Plast) with a 30 mm headspace (H1) and a 50 mm headspace (H2). The film was LINTOP PE HB A40 (Linpaq). The pH of meat were measured using a Sydel pH meter and a lot406 Metter Toledo electrode. Analysis of gas (% O2 and % CO2) were also 259

mailto:*[email protected]


performed (Checkpoint II, PBI Dansensor). Sensory analyses were performed using a panel of French consumers who evaluated the products in the trays and after cooking on a pan and served at 64°C. Case-ready packagings (n=250) were stored in the dark at 4°C for 9 days in a cold room. The microbiological evolutions of the lactic acid bacteria (NF ISO 15214), enterobacteriacae (NF ISO 21528-2), Pseudomonas (NF V04-504), Brochothrix thermosphacta (NF V04-505) and Aerobic colony count (NF EN ISO 4833) were monitored. At each point of analysis (days 2, 5, 9), bacterial counts were performed on five samples of each batch. Those products (n=250) were placed for 1/3 of their microbiological shelf-life at 4°C and for 2/3 at 8°C in a cold room. Statistical analysis were conducted with 9.2 SAS software version (SAS Institute, USA) using ANOVA, the Turkey and Student tests.

RESULTS AND DISCUSSION

Impact of the headspace We followed the composition of CO2 and O2 in each trays to investigate the effect of headspace on the composition of gas. The 50/50 gas follows the same pattern for the two headspaces (figure 1). The CO2 dissolution in the water-phase and the fat-phase of the pork chop is observed during the first hours of storage. Metmyoglobin formation and microbial development are responsible for oxygen consumption and could influence gas composition after two days of storage. Gas composition follow the same trend that was estimated by Bjørn et al (2007) with his mathematical model. With the same initial gas injection, the estimated gas equilibrium reaches here 35.2% CO2 and 58.5% O2.

Fig. 1. Gas composition evolution in two different

headspaces (n=250)

The dissolution of CO2 during the first period of storage is influenced by temperature and pH values. Gill (1988) showed a significant effect of temperature, type of fat tissue and fat content on the solubility of CO2 in meat. Lower solubility at higher temperatures has been mentioned by many authors (Wolfe, 1980; Wimpfheimer et al., 1990; Zhao et al., 1994) as the reason for the minor effect of MAP at higher temperatures. In our study, pork chops were preserved at 4°C. According to Devlieghere (1998), an increase in pH resulted in an increase of the CO2 solubility and this effect was strongest at high pH values. Similar results were demonstrated by Löwenadler et al (1994). In our study pH measures were constant and homogenous during storage as we can see on the table 1. Table 1 pH values in trays H1 and H2 with 50% O2 and 50% CO2

50/50 H1 (n=125) 50/50 H2 (n=125)

D2 5,7±0.3 5,7±0.2

D5 5,7±0.3 5,7±0.1

D9 5,6±0.3 5,7±0.2

Table 2 shows that the level of contamination of Enterobacteriacae at day 10 is significantly lower for small trays (H1) versus large sizes (H2) cf. table 2.

Table 2 Contamination level of Enterobacteriacae in two headspaces

Enterobacteriacae Log cfu/cm² at 10 days (n=85)

Headspace Average Level (5%) *

H1 0.19 A

H2 1.01 B

*a,b Proportion within a column sharing the same letter are not significantly different (p>0,05).

Whatever the floras and dates, there are no significant differences between H1 and H2 trays; this means that the headspace H1 allowed an equivalent conservation as H2. We can conclude that it’s possible to reduce the volume of gas and headspace whatever the gas mixture used. When observing trays (table 3), consumers prefer smaller trays (small headspace). The best purchase intent (78%) is obtained with the lowest oxygen content (40%) gas. It may be noted that the most

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frequent used by and the pork meat industry (gas 70/30) is also very popular in a package with a reduced headspace with 74% purchase intent before opening. Table 3 Evaluation of the trays by a panel of consumers

Gas/headspace Note /10 Percentage of consumers satisfied

40/60H1 7.0a 65% 70/30H1 6.9a 63% 60/40H1 6.7ab 65% 50/50H1 6.5ab 49% 50/50H2 6.2abc 46% 40/60H2 6.2abc 42% 70/30H2 5.9bc 39% 60/40H2 5.5c 34%

*a, b Proportion within a column sharing the same letter are not significantly different (p>0,05).

Impact of the gas compositions: We followed the microbial evolution during the shelf life and the pork chops were evaluated by consumers in order to investigate the effect of gas composition. After 10 days of storage (table 4) gas composition shows a significant effect on microbial growth for Pseudomonas, Enterobacteriaceae, and Brochothrix. Table 4 Comparison of contaminations after 10 days of storage Log cfu/cm² at 10 days (n=85) Pseudomonas Entero-

bacteriacae Brochothrix

O2/CO2 Average Average Average 40/60 1.68a 0.05a 2.57a 50/50 2.69ab 0.49ab 2.71a 60/40 2.47b 0.64ab 3.48b 70/30 3.56c 1.21b 4.22c


In the same thermal scenario there are 3.56 versus 1.68 log cfu/cm² of Pseudomonas with gas 70/30 compared to 40/60. The growth of Enterobacteriacae is reduced of 1 log and 2 log of Brochothrix with 60% of CO2 versus 30%. The growth of bacteria is quite similar with 60 and 50% of CO2. The effect of modified atmosphere packaging is not surprising can mainly be attributed to the bacteriostatic action of CO2. Devlieghere (1998) reported that the growth of P. fluorescens was highly influenced by CO2. A significant increase in the lag phase and a decrease in the growth rate of P. fluorescens was, as could be expected, noticed with increasing CO2 concentrations. The figure 1 represents of growth of aerobic colony count during ten days for all

gases and headspaces. According to the microbial results (fig. 2), the shelf life of pork chops may vary between gas compositions.

Fig. 2 The growth of aerobic colony count during ten

days (log cfu/cm²) After 7 days of storage at 4°C, pork chop were evaluated by 65 consumers. Table 5 Tasting session by a panel of consumers

Products

Overall impression (n=250) Re-consume Intention Note /10

Percentage of consumers satisfied

RI

70/30H1 6.9a 63% 74% a 70/30H2 6.6ab 55% 65% ab 50/50H2 6.4ab 51% 55% b 60/40H2 6.4ab 48% 63% ab 50/50H1 6.3ab 52% 62% ab 40/60H1 6.1b 46% 57% b 60/40H1 5.9b 42% 54% b 40/60H2 5.7b 48% 51% b


At the tasting, gas with the highest CO2 level is found ranked lower. Clearly, the gas mixture commonly used is the most appreciated by the consumer panel when combined with a small headspace. The intention to re-consume the product is statistically equivalent between the reference sample (70/30 H2) and the product (50/50 H1). Consumers noticed dryness of the meat in the (40/60) gas composition. Most of the consumers panel were satisfied by the gas mixture (70/30) with the small headspace.

Validation of shelf life According to the experimental results the trays with the lowest headspace enable a microbial preservation at least equivalent to those with large headspace. Furthermore they are more appreciated by consumers. The choice of the gas mixture was oriented towards the gas having the highest rates of CO2, because they provide a longer shelf life. With the aim of increasing the shelf life of pork chops. Two batches of trays packed under two gas compositions 40/60 H1 and 50/50 H1 products

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were monitored (evolution of gas and growth of bacteria) on two scenarios:

Scenario 1: 4°C until D8, 8°C until D12 Scenario 2: 4°C until D9, 8°C until D14

Table 6 Evolution of contaminations of several floras in two scenarios of temperature

Log cfu/cm² 40/60 H1 Log cfu/cm² 50/50 H1

Days Pseud. Ent ACC List. Pseud. Ent ACC List. D1 (n=5) 1.86 1.00 2.39 Abs 2.00 1.06 2.22 Abs D8 (n=5) 2.25 1.75 4.48 Abs 2.38 1.63 4.58 Abs

D12 (n=5) 2.43 2.60 6.60 Abs 3.18 3.08 6.65 Abs D9 (n=5) 2.22 1.53 4.99 Abs 2.00 1.33 4.38 Abs

D14 (n=5) 3.08 2.21 6.49 Abs 3.51 3.51 6.83 Abs

This observation means that within the two scenarios for shelf life evaluations, the Pseudomonas concentration obtained under gases (40/60 and 50/50) at the end of the shelf life remains acceptable according to the GHP guide criterion (6 log cfu/cm²). In the same way Enterobacteriacae remains acceptable according to the GHP guide criterion (5 log cfu/cm²). These results confirm a shelf life of 12 and 14 days for your individual retail packs, 4 days longer than the SL currently positioned on the individual retail packs.

CONCLUSION This study confirms that a reduction of the O2 percentage of oxygen from 70% to 40% in the gas mixture doesn’t impact the color of the pork meat for consumers. The increase of the CO2 percentage extends the shelf life of MAP. Consumers are sensible to the volume reduction of the trays. This reduction of headspace can lower the environmental impact without degrading the sanitary quality of the meat.

ACKNOWLEDGEMENTS The authors thank all the partners of the projects ATMO (Idele, Oniris, Air liquide, Elivia, Adria développement). This work was supported by the Area of Brittany.

REFERENCES

Paper: 1. R.A. Mancini, M.C. Hunt (2005). Current

research in meat color. Meat Science 71 100-121.

2. M. Jakobsen, G. Bertelsen (2000). Colour stability and lipid oxidation of fresh beef. Development of a response surface model for predicting the effects of temperature, storage time, and modified atmosphere composition. Meat Science, 54, 49-57.

3. Bjørn Tore Rotabakk, John Wyller, Odd Ivar Lekang b, Morten Sivertsvik (2007). A mathematical method for determining equilibrium gas composition in modified atmosphere packaging and soluble gas stabilization systems for non-respiring foods. Journal of Food Engineering 85 479-490 23.

4. W. Kenneth, McMillin (2008). Where is MAP Going? A review and future potential of modified atmosphere packaging for meat. Meat Science 80 43-65.

5. C.O. Gill, N. Penney (1988). The effect of the initial gas volume to meat weight ratio on the storage of chilled beef packaged under carbon dioxide. Meat Science 22 53-63.

6. S.K.Wolfe (1980). Use of CO and CO2

enriched atmospheres for meats, fish and produce. Food Technol. 34, 55-58.

7. L.Wimpfheimer, N.S. Altman, Hotchkiss, J.H (1990). Growth of Listeria monocytogenes Scott A, serotype 4 and competitive spoilage organisms in raw chicken packaged under modified atmospheres and in air. Int. J. Food Microbiol. 11, 205-214.

8. Y. Zhao, J.H.Wells, K.W. McMillin (1994). Applications of dynamic modified atmosphere packaging systems for fresh redmeats: a review. J. Muscle Foods 5, 299-328.

9. F. Devlieghere, J. Debevere, J. Van Impe (1998). Concentration of carbon dioxide in the water-phase as a parameter. International Journal of Food Microbiology 43 105-113.

10. J. Löwenadler, U. Rönner, (1994). Determination of dissolved carbon dioxide by coulometric titration in modified atmosphere systems. Lett. Appl. Microbiol. 18, 285-288.

Book Chapter : 1. A. O Gill & C. O. Gill, (2005). Preservative

packaging for fresh meats, poultry, and fin fish. In J. H. Han (Ed.), Innovations in food packaging (pp. 204-226). Amsterdam: Elsevier Academic Press.

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60th International Congress of Meat Science and Technology, 17-22rd August 2014, Punta del Este, Uruguay

LOSSES DUE AGING PROCESS IN BREAST MEAT OF BROILERS WITH DIFFERENT GENDERS AND AGES

Juliana L. M. Mello1*, Ana Beatriz B. Rodrigues1, Aline Giampietro-Ganeco1,

Fábio B. Ferrari1, Hirasilva Borba1, Rodrigo A. de Souza1 and Pedro A. de Souza1 1Department of Technology, Faculty of Agrarian and Veterinary Sciences, São Paulo State University, UNESP Jaboticabal, São Paulo, Brazil.

*[email protected]

Abstract – The aim of this study was to evaluate the possible losses due aging process, related to the quality of breast meat in broilers of fast growth lineage. Pectoralis major muscle from deboned carcasses was purchased in a commercial slaughterhouse and used in this study. A completely randomized design in 2x2x3 factorial was used with two ages, two genders and three aging periods in ten replications. The meat of birds in slaughter age (42 days old) had higher storage weight loss, higher exudate volume and lower soluble protein. The gender of birds influenced the storage weight loss and the quantity of soluble protein present in the exudate fluid. The meat of males showed higher storage weight loss and lower soluble protein. The lipid oxidation was not influenced by age and gender of birds. The longer the aging period, the higher is the storage weight loss, the exudate volume and the lipid oxidation of samples. The age, gender of birds and time of aging affect the storage weight loss of meat as well as the quantity of soluble protein lost in the exudate fluid. The increase of the aging period favors the lipid oxidation in the meat of broilers. I. INTRODUCTION With advancing age, lots of cocks and hens are discarded by showing a rigid meat, mainly due to the increased collagen concentration. Females are usually large birds, weighing approximately 3-4 Kg, whose body shape is similar to a broiler, with very developed breast muscles and legs [1]. However, they have a large fat deposition, both subcutaneous and abdominal and the meat is tougher and less juicy. Due to low consumer preference for this meat because of their sensory quality, it becomes necessary to seek alternative techniques, such as maturation, that add value and enable consumption of this meat as prime beef and not as by-products. Thus, the aim of this study was to evaluate the possible losses due aging process regarding to breast meat quality of broilers.

II. MATERIALS AND METHODS This study was developed in Technology Laboratory of Animal Products in São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil. Carcasses from 120 broilers of fast growth lineage, males and females with 42 days of age and spent birds (approximately 70 weeks of age) purchased from a commercial slaughterhouse were used in this study. After deboned, samples of the Pectoralis major muscle (breast) were individually packed in vacuum and stored in a BOD chamber, under ± 2°C for 3 and 7 days. Storage weight loss, exudate volume, soluble protein in the exudate and lipid oxidation were analyzed. The storage weight loss was determinate by difference between initial and final weight, before and after aging, expressed as percentage. The dosage of the soluble protein in the exudate was performed according Hartree [2]. Lipid oxidation was evaluated by measuring the substances reactive to thiobarbituric acid-2 (TBARS) according to the methodology proposed by Pikul et al. [3]. For statistical analysis a completely randomized design in 2x2x3 factorial was used with two ages (broilers at 42 days of age and spent birds at approximately 70 weeks), two genders and three aging periods, in ten replications. Data were submitted to analysis of variance and means compared by Tukey test (5%) using the statistical program SAS [4]. III. RESULTS AND DISCUSSION

Table 1 shows the average results obtained from storage weight loss, exudate volume, soluble protein in the exudate and lipid oxidation analysis of broilers and spent birds meat, subjected to aging process for seven days. The age of birds influenced storage weight loss, the exudate volume and the quantity of soluble protein present in the exudate fluid. The meat of

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birds in slaughter age (42 days old) showed higher storage weight loss, higher exudate volume and lower soluble protein.

Table 1 Storage weight loss (%), exudate volume (mL/g sample), soluble protein in the exudate (mg protein/mL drip) and lipid oxidation (mg MDA/Kg

sample) in breast meat of broilers with different genders and ages

SL Exudate SP TBARS

Age (A)

42 days 2.95 A 0.025 A 0.154 B 0.135

70 weeks 1.50 B 0.010 B 0.174 A 0.134

Gender (G)

Male 2.31 A 0.018 0.160 B 0.134

Female 2.14 B 0.017 0.168 A 0.135

Aging periods (T)

Control group

- - - 0.116 C

3 days 1.66 B 0.012 B 0.179 A 0.138 B

7 days 2.78 A 0.023 A 0.149 B 0.149 A

P-value (A) <0.0001 <0.0001 <0.0001 0.6664

P-value (G) 0.0042 0.0569 <0.0001 0.5618

P-value (T) <0.0001 <0.0001 <0.0001 <0.0001

CV (%) 11.61 15.60 4.18 8.33

Averages followed by different letters differ according to Tukey’s test. The following abbreviations are used: SL: Storage weight loss; SP: Soluble protein in the exudate; TBARS: lipid oxidation; CV: coefficient of variation; The gender of birds influenced the storage weight loss and the quantity of soluble protein present in the exudate fluid. The meat of males showed higher storage weight loss and lower soluble protein. The lipid oxidation was not influenced by age and gender of the birds. The aging process influenced these variables. As aging period increases, the storage weight loss also increases due to greater loss of exudate fluid. There was an increase in lipid oxidation. Bowker and Zhuang [5] evaluating the relationship between the composition of exudate liquid and breast meat quality of broilers, concluded that this composition and volume may change depending on pH, color and water holding capacity of meat. Vaithiyanathan et al. [6] studying the aging process of breast meat from spent hens with 72 weeks of age, observed the increase in TBARS value over the time.

Lipid oxidation is an important reaction which limits the shelf life of foods, it is one of the primary mechanisms of meat quality deterioration, especially for samples with higher quantity of fat. The storage of meat under low temperatures, such as during the aging process, can minimize lipid oxidation, however, even in freezing temperatures this is just delayed but not eliminated [7]. During storage the breast meat had a significant increase in the volume of exudate fluid. Roça [8] reports that during the meat aging there is slight increase in water holding capacity due to the higher pH and enzymatic degradation of myofibrillar structure. Thus, the exudate volume decreases and, consequently, decreases the storage weight loss, in contrast to what was observed in this study. IV. CONCLUSION

The age, gender of birds and the time of aging affect the storage weight loss of meat as well as the quantity of soluble protein lost in the exudate fluid. The increase of the aging period favors the lipid oxidation in broiler meat. ACKNOWLEDGEMENTS This study was financially supported by Foundation for Research Support in the State of São Paulo (Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP, 2012/08787-7 and 2011/21681-0) to whom the authors want to express their gratitude. REFERENCES 1. Sanfelice, C., Mendes, A. A., Komiyama, C. M.,

Cañizares, M. C., Rodrigues, L., Cañizares, G. I. L. (2010). Avaliação do efeito do tempo de desossa sobre a qualidade da carne de peito de matrizes pesadas de descarte. Acta Scientiarum. Animal Sciences. 32:85-92.

2. Hartree, E. F. (1972). Determination of protein: a modification of the Lowry method that gives a linear photometric response. Analytical Biochemistry. 48:422-427.

3. Pikul, J. (1989). Evaluation of three modified TBA methods for measuring lipid oxidation in chicken meat. Journal of Agriculture and Food Chemistry. 37:1309-1313.

4. SAS Institute. (2002). SAS user’s guide: statistics. Release 9.1. Cary.

5. Bowker, B. C., Zhuang, H. (2013). Relationship between muscle exudate protein composition

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and broiler breast meat quality. Poultry Science, 92(5):1385-1392.

6. Vaithiyanathan, S., Naveena, B. M., Muthukumar, M., Girish, P. S., Ramakrishna, C., Sen, A. R. and Babji, Y. (2008). Biochemical and Physicochemical Changes in Spent Hen Breast Meat During Postmortem Aging. Poultry Science, 87(1):180-186.

7. Mariutti, L. R. B.; Bragagnolo, N. (2009). A oxidação lipídica em carne de frango e o impacto da adição de sálvia (Salvia officinalis, L.) e de alho (Allium sativum, L.) como antioxidantes naturais. Revista do Instituto Adolfo Lutz, 68:1-11.

8. Roça, R. O. (2002). Tecnologia da carne e produtos derivados. Botucatu: Faculdade de Ciências Agronômicas UNESP, 202 p.

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ATTRIBUTES RELATED TO TENDERNESS IN BREAST MEAT OF BROILERS WITH DIFFERENT GENDERS AND AGES AFTER AGING

Juliana. L. M. Mello1*, Ana Beatriz B. Rodrigues1, Aline Giampietro-Ganeco1,

Fábio B. Ferrari1, Hirasilva Borba1, Leonardo D. C. Vieira1 and Rodrigo A. de Souza1 1Department of Technology, Faculty of Agrarian and Veterinary Sciences, São Paulo State University, UNESP Jaboticabal, São Paulo, Brazil.

*[email protected]

Abstract – The aim of this study was to evaluate the attributes related to tenderness of breast meat in broilers of fast growth lineage with two different genders and ages, after aging. Pectoralis major muscle from deboned carcasses was purchased in a commercial slaughterhouse and used in this study. A completely randomized design in 2x2x3 factorial was used with two ages, two genders and three aging periods, in ten replications. As the age of birds increases, there is a reduction of cooking weight loss, an increase of collagen percentage and, consequently, increased shear force. The meat from males showed greater cooking weight loss, higher collagen percentage and therefore increased shear force. The aging process promoted an increase of cooking weight loss and reduction of the collagen percentage and shear force, after three days of storage. The variables studied showed no significant difference between three and seven days of storage. The age of bird reduces cooking weight loss, increases the collagen content and consequently affects tenderness of breast meat. The gender of broilers influences the cooking weight loss, collagen percentage and tenderness of breast meat. The aging process favors the meat softening from spent birds. I. INTRODUCTION The aging of meat is a period during which significant changes in quality parameters and microstructure of muscle, especially related to texture, tenderness, water holding capacity and sensory properties of meat so that the muscle to meat conversion occurs properly, significantly influencing the palatability [1]. After slaughter, meat of spent birds is usually used in the formulation of emulsified sausages, burgers, among others, and may also be sold at a lower price than conventional chicken, a viable way to give the appropriate destination at the end of production and add value to this unexplored poultry segment. Thus, the aim of this study was to evaluate some attributes related to tenderness of broilers and spent birds meat after aging for up to seven days, so that aging process can be used as a

softening technique that adds value for this type of product. II. MATERIALS AND METHODS This study was developed in Technology Laboratory of Animal Products in São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil. Carcasses from 120 broilers of fast growth lineage, males and females with 42 days of age and spent birds (approximately 70 weeks of age) purchased from a commercial slaughterhouse, were used in this study. After deboned, samples from Pectoralis major muscle (breast) were individually vacuum packed and stored in a BOD chamber, under ± 2°C for 3 and 7 days. Cooking weight loss, shear force and collagen percent were analyzed. The cooking weight loss was determined by cooking the samples in a water bath according to methodology described by Honikel [2]. From cooked samples subsamples with known area (cm²) were obtained and submitted to cut in Texture Analyser TA-XT2i texturometer coupled to Warner Bratzler device, which determined the shear force in kgf. The collagen percent was determined by quantification of hydroxyproline amino acid according to the methodology proposed by Woessner Junior [3]. For statistical analysis a completely randomized design in 2x2x3 factorial was used with two ages (broilers at 42 days of age and spent birds at approximately 70 weeks), two genders and three aging periods, in ten replications. Data were submitted to analysis of variance and means compared by Tukey test (5%) using the statistical program SAS [4]. III. RESULTS AND DISCUSSION

Table 1 shows the average results obtained from cooking weight loss, shear force and

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collagen percent analysis of broilers and spent birds meat, subjected to aging process for up to seven days. It is observed that with increasing of bird age, there is a reduction of cooking weight loss because meat from older birds has higher water holding capacity. The meat from spent birds showed an increase of collagen percentage and, consequently, increased shear force, reducing the tenderness.

Table 1 Cooking weight loss (CWL), shear force (SF) and collagen percent in breast meat of broilers

with different genders and ages

CWL (%)

SF (kgf/cm²)

Collagen (%)

Age (A)

42 days 29.90 A 2.194 B 3.74 B

70 weeks 27.91 B 2.669 A 5.44 A

Gender (G)

Male 30.18 A 2.665 A 5.36 A

Female 27.63 B 2.198 B 3.82 B

Aging periods (T)

Control group 26.81 B 3.489 A 4.97 A

3 days 29.98 A 1.928 B 4.29 B

7 days 29.92 A 1.877 B 4.51 B

P-value (A) 0.0001 <0.0001 <0.0001

P-value (G) <0.0001 <0.0001 <0.0001

P-value (T) <0.0001 <0.0001 <0.0001

CV (%) 9.39 11.53 12.52

Averages followed by different letters differ according to Tukey’s test. The following abbreviations are used: CV: coefficient of variation; The meat from males showed greater cooking weight loss, higher collagen percentage and therefore increased shear force. The aging process promoted an increase of cooking weight loss and reduction of the collagen percentage and shear force, after three days of storage. The variables studied showed no significant difference between three and seven days of storage. According Hadlich et al. [5] the gender influences collagen content, while males have a higher amount of intramuscular connective tissue than females. Roça [6] reports that during the aging process of meat occurs a slight increase in water holding capacity due to the higher pH and enzymatic degradation of myofibrillar

structure. Thus, the exudate liquid volume decreases and, consequently, the storage weight loss. However, an increase in the water holding capacity does not prevent the increase of cooking weight loss, which possibly due to relaxation of muscle fibers during aging process. According Vaithiyanathan et al. [7], during aging of meat from spent hen breast, there are many changes in the structural proteins degradation and tenderization. These changes in the structural proteins improved the tenderness of spent hen breast meat stored at low temperatures. This improvement in tenderness through postmortem aging may be considered as an option to add market value to spent hen breast muscle. IV. CONCLUSION

The age of bird reduces cooking weight loss, increases the collagen percent and consequently affects tenderness of breast meat. The gender of broilers influences the cooking weight loss, collagen percent and tenderness of breast meat. The aging process favors the softening of meat from spent birds. ACKNOWLEDGEMENTS This study was financially supported by Foundation for Research Support in the State of São Paulo (Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP, 2012/08787-7 and 2011/21681-0) to whom the authors want to express their gratitude. REFERENCES 1. Komiyama, C. M., Martins, M. R. F. B..

Mendes, A. A., Sanfelice, C., Cañizares, M. C. S., Roça, R. O., Almeida, I. C. L. (2009). Avaliação da técnica de maturação sobre a qualidade da carne e estrutura da fibra muscular do peito de matrizes pesadas de descarte de frangos de corte. Brazilian Journal of Food Technology. v.2.

2. Honikel, K. O. (1987). The water binding of meat. Fleischwirttsch. 67:1098-1102.

3. Woessner Junior, J. F. (1961). The determination of hydroxyproline in tissue and protein samples containing small proportions of this amino acid. Archives of Biochemistry and Biophysics. 93:440-447.

4. SAS Institute. (2002). SAS user’s guide: statistics. Release 9.1. Cary.

5. Hadlich, J. C., Longhini, L. G. R., Mason, M. C. (2008). A influência do colágeno na textura

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da carne. Pubvet, v.2. Available at: <http://www.pubvet.com.br/artigos_det.asp?artigo=160> Accessed on: Feb. 24, 2012.

6. Roça, R. O. (2002). Tecnologia da carne e produtos derivados. Botucatu: Faculdade de Ciências Agronômicas UNESP, 202 p.

7. Vaithiyanathan, S., Naveena, B. M., Muthukumar, M., Girish, P. S., Ramakrishna, C., Sen, A. R., Babji, Y. (2008). Biochemical and physicochemical changes in spent hen breast meat during postmortem aging. Poultry Science. 87:180-186.

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EFFECT OF LACTATE AND DIACETATE SALTS AND HIGH PRESSURE PROCESSING ON THE SURVIVAL OF LISTERIA

MONOCYTOGENES IN CURED BEEF CARPACCIO

Yanina X. Barrio1,2*, Diego Wilches Perez3, Sergio R. Vaudagna1,2,4 and Jordi Rovira3

1 Instituto Tecnología de Alimentos, CIA, INTA, Morón, Buenos Aires, Argentina

2 Facultad de Ingeniería y Ciencias Exactas, UADE, Buenos Aires, Argentina

3 Departamento de Biotecnología y Ciencia de los Alimentos, Universidad de Burgos, Burgos, España

4 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina

*[email protected]

Abstract – The behavior of Listeria monocytogenes strains and spoilage microflora, in response to the addition of antimicrobial additives (potassium lactate 3.0% or potassium lactate 1.7% and sodium diacetate 0.12%) and high pressure processing (HPP), was evaluated on cured beef carpaccio during refrigerated storage. Samples with or without antimicrobials were inoculated with a five-strain pool of L. monocytogenes (at ca. 104 cfu/g), frozen at -60°C and pressurized at 600 MPa for 5 min at room temperature. Afterwards, samples were stored at 4°C for up to 21 days. HPP alone was sufficient to reduce the L. monocytogenes count below the detection limit (<2 log cfu/g). Concerning antimicrobials addition, potassium lactate and sodium diacetate contributed to delay the growth of lactic acid bacteria and psycrothophs of pressurized samples during refrigeration storage. I. INTRODUCTION The commercialization of a wide variety of "ready-to-eat" (RTE) meat products, including fresh marinated or cured meats has been increasing in the international and local markets. Beef carpaccio is a traditional Italian dish prepared as thin slices from frozen raw or cured beef, which are packed under vacuum or modified atmospheres, and kept at refrigeration temperature until consumption. Because of the minimal process that carpaccio is subjected, and the possibility of contamination with pathogenic bacteria, such as Escherichia coli O157:H7, Salmonella and Listeria monocytogenes, carpaccio is considered a high-risk food (1). In addition, conventional preservation technologies like thermal pasteurization cannot be applied because of the detrimental effects on product’s appearance and sensory characteristics. In this sense, the use of a non-thermal technology such as High Pressure could be an alternative for carpaccio pasteurization. High Pressure Processing (HPP) has been successfully applied for the pasteurization of dried or cooked meat

products (2). However, HPP of fresh pigmented meats causes a significant discoloration at pressure levels required for vegetative cells inactivation (>300 MPa) (3). Some studies (4, 5) have reported that HPP applied to frozen carpaccio improved the appearance of pressurized red meats but these treatments resulted less effective to microbial inactivation. HPP combined with natural antimicrobials has been proved in some foodstuffs (6). In this concern, lactate salts have been widely used by meat industry to extend shelf life of products. Also, some studies have shown a synergistic effect of lactate and diacetate combination against several foodborne pathogens (7). The aim of this study was to evaluate the combined effect of lactate and diacetate salts and HPP on the survival of Listeria monocytogenes and spoilage microflora of cured beef carpaccio. II. MATERIALS AND METHODS Bacterial strains and inocula preparation Listeria monocytogenes strains used in this study included five isolates from the International Life Sciences Institute North America (ILSI NA) L. monocytogenes strain collection (ILSI no. 7, 11, 29, 35 and 36) (8). The strains were maintained in frozen culture at -80°C until subcultures were prepared by inoculating test tubes with 5 ml of Brain Heart Infusion broth (BHI, Oxoid, England) with a single colony growing in Triptone Soya Agar (TSA, Oxoid, England) supplemented with 0.3% (w/v) yeast extract and 0.1% (w/v) of sodium pyruvate, and incubated at 37°C overnight. The cultures were centrifuged at 7000 rpm at 4°C for 10 min and the pellets washed twice with Ringer solution (Oxoid, England). Then, the suspensions were diluted three times in 9 ml of Ringer solution (Oxoid, England) to give an approximate concentration of 106 cfu/ml. The pool was prepared by mixing equal volumes of each strain.

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Carpaccio samples preparation and HPP Bovine Semitendinosus muscles were rubbed in by hand into plastic bags (Cryovac HT, Cryovac L.T., Spain) with curing salts (weight of product basis: sodium chloride 1.2%, sodium tripolyphosphate 0.1%, sodium citrate 0.05%, sodium nitrite 0.015%, sodium isoascorbate 0.05%) or with curing salts plus the addition of potassium lactate 3.0% (PURASAL Hi-Pure P Plus, PURAC, Spain) or potassium lactate 1.7% and sodium diacetate 0.12% (PURASAL Opti.Form PD4, PURAC, Spain). Then, muscles were vacuum-packed and stored at 4°C for 12 days. After chill storage, cured muscles were frozen and stored at -20°C until slicing (1.5-2 mm). Carpaccio slices (25±1 g) were inoculated with the pool of L. monocytogenes strains to obtain a concentration of approximately 104 cfu/g. Then, samples were vacuum-packed (Cryovac HT, Cryovac L.T., Spain) and frozen at -60ºC. Frozen carpaccio samples were pressurized at 600 MPa for 5 min in a discontinuous hydrostatic pressurization unit (Hiperbaric 135, Hiperbaric, Burgos, Spain). The initial water (pressurization fluid) temperature was 15°C. After treatment, all samples were stored at 4°C and analyzed after 1, 7, 14 and 21 days of storage. Experimental design and statistical analysis The experiment was arranged in a complete randomized 3x2 factorial design. The model included the main effects of antimicrobial addition (none, potassium lactate 3.0% or potassium lactate 1.7% and sodium diacetate 0.12%), HPP (0.1 or 600 MPa) and antimicrobial by HPP interactions. Three experimental units were used for each treatment. Then, ANOVA was performed to evaluate significant (p<0.05) effect of factors and the Tukey multiple comparisons test (p<0.05) was applied to analyze statistical differences among means. Statistical analysis was performed using the SPSS-v12 software (SPSS, Chicago, IL., USA). Microbiological and chemical analyses For microbiological analysis, carpaccio slices were aseptically removed from their plastic wrapping and stomached (AES laboratoire, France) for 1 min in sterile bags with filter (Bag Page, Interscience, France) with sufficient volume of Ringer solution (Oxoid, England) to obtain an initial 1/10 (w/v) dilution. Successive decimal dilutions were then performed and

plated onto appropriate medium. L. monocytogenes were determined by plate counting in Chromogenic Listeria Agar (Oxoid, England) supplemented with OCLA (ISO) Selective Supplement (SR0226E, Oxoid, England) and Brillance™ Listeria Differential Supplement (SR0228E, Oxoid, England), after 48 h of incubation time at 37ºC. Lactic acid bacteria (LAB) were determined by plating on Man, Rogosa and Sharpe agar (MRS, Oxoid, England) incubated for 72 h at 30ºC, while psychrotrophs on Plate Count Agar (PCA, Merck, Germany) incubated for 10 days at 8°C. The water activity (aw) and pH was determined for each carpaccio formulation. Water activity was measured at 25°C using a water activity meter (AquaLab CX-2, Decagon Devices Inc., USA) and pH using a digital pH-meter (Micro pH 2001, Crison, Barcelona, Spain). III. RESULTS AND DISCUSSION Water activity (aw) and pH In regard to carpaccio formulation, the aw values of samples with curing salts or with curing salts plus potassium lactate or potassium lactate and sodium diacetate were 0.978±0.003, 0.970±0.001 and 0.973±0.003, respectively. In addition, the pH values were 5.47±0.03, 5.80±0.03 and 5.78±0.02, respectively. Microbiological counts Table 1 shows the microbial evolution on carpaccio samples treated with or without antimicrobials and HPP, during 21 days of storage at 4°C. A significant (p<0.05) HPP effect was observed in L. monocytogenes counts. It can be seen that HPP was able to reduce the L. monocytogenes count below the detection limit (<2 log cfu/g). Moreover, L. monocytogenes remained undetectable in all pressurized samples throughout the storage. Regarding unpressurized samples (0.1 MPa), no significant (p>0.05) differences were observed in L. monocytogenes counts of samples with or without antimicrobial addition, at each day of storage. HPP has been recognized by the Food Safety and Inspection Service of USA as a useful listericidal post-processing and post-packing treatment for RTE foods (9). In addition, several studies reported about the effectiveness of HPP to control L. monocytogenes in RTE meat products. In this sense, Jofré et al. (10) reported that pressurization of sliced cooked ham at 600 MPa

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Table 1. Microbial evolution (log cfu/g) of carpaccio samples treated with or without antimicrobials and high pressure, during storage at 4ºC.

Antimicrobial*

Pressure (MPa)

Days of storage

1 7 14 21

Listeria monocytogenes

None 0.1 4.31±0.20 ns 4.00±0.30 ns 5.08±0.84 ns 3.70±0.20 ns 600 <2 <2 <2 <2 PL 0.1 5.02±0.18 ns 5.38±0.64 ns 4.47±0.47 ns 3.70±0.15 ns 600 <2 <2 <2 <2 PL+SD 0.1 4.37±0.51 ns 4.10±0.17 ns 4.37±0.65 ns 3.95±0.30 ns 600 <2 <2 <2 <2

LAB None 0.1 6.06±0.07 A 7.34±0.17 a 6.67±0.58 a 7.30±0.35 a 600 2.05±0.95 B 4.81±0.40 b 6.18±0.34 b 7.01±0.41 a PL 0.1 5.56±0.08 A 6.52±0.05 a 6.36±0.18 a 6.33±0.06 ab 600 2.52±0.24 B 3.89±0.04 bc 5.43±0.02 bc 6.31±0.72 ab PL+SD 0.1 5.67±0.20 A 6.47±0.11 a 6.39±0.36 a 6.67±0.02 b 600 <2 3.40±0.45 c 5.29±0.10 c 5.16±0.45 b Psycrotrophs None 0.1 7.00±0.20 A 8.08±0.09 a 8.43±0.08 ns nd 600 4.00±0.18 B 5.84±0.27 c 7.46±1.03 ns 8.28±0.05 ns PL 0.1 6.00±0.10 A 7.52±0.08 b 7.30±0.30 ns nd 600 3.00±0.24 B 4.41±0.15 d 7.35±0.14 ns 8.08±0.27 ns PL+SD 0.1 6.00±0.12 A 7.45±0.10 b 7.77±0.07 ns nd 600 3.00±0.30 B 3.86±0.29 e 7.37±0.44 ns 7.82±0.27 ns

*Antimicrobial: none, potassium lactate 3.0% (PL) and potassium lactate 1.7% and sodium diacetate 0.12% (PL+SD). Means with different capital letters in the same column indicate significant differences (p<0.05) regarding pressure effect while means with different lower case letters in the same column indicate significant differences (p<0.05) regarding antimicrobial by pressure interaction effect. ns: non-significant / nd: non-determined. for 5 min at 10ºC produced a significant reduction of around 3.5 log cfu/g in L. monocytogenes counts and samples showed levels lower than 10 cfu/g during 90 days of storage at 1ºC and 6ºC. Also, Koseki et al. (11) observed that the number of L. monocytogenes on cooked ham inoculated with 105 cfu/g was initially reduced by HPP at 500 MPa for 10 min to below the detectable level (10 cfu/g). However, the bacterial count gradually increased during storage at 10ºC, and exceeded the initial inoculum level at the end of the 70-day period. The analysis of the spoilage microflora of carpaccio showed an initial population of LAB and psycrotrophs between 5.5 and 7 log cfu/g depending on carpaccio formulation. After HPP (day 1), both microbial counts were significantly (p<0.05) reduced by about 3 log cycles. Garriga et al. (6) report that marinated beef loin slices treated at 600 MPa for 6 min at 31ºC showed a significant reduction of at least 4 log cycles after treatment for aerobic, psycrothrophic, and LAB counts. Instead,

Realini et al. (12) informed that frozen cured pork carpaccio treated at 600 MPa for 6 min at freezing temperature (−15°C or −35 °C) had only 0.5 and 1.5 log cycle reductions of LAB and psychrotrophs counts, respectively. Regarding antimicrobial addition, significant (p<0.05) interaction of HPP and antimicrobial additives was observed in LAB and psycrotrophs counts during storage at 4ºC. Samples with potassium lactate 1.7% and sodium diacetate 0.12% treated at 600 MPa presented a delay in the growth of LAB and the lowest counts (5.16 log cfu/g) at 21 days of storage. This effect was also observed in psycrotrophs count of pressurized samples with potassium lactate 3.0% and with potassium lactate 1.7% and sodium diacetate 0.12% at day 7 of storage. However, psycrothophs counts shown a noticeable increase at day 14 and no significant (p>0.05) differences were observed among treatments. In this concern, Diez et al. (13) observed that the addition of 3% mixture of potassium/sodium L-lactate to morcilla de Burgos formulation extended the shelf life by

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around three weeks respect to control samples but the combination of HPP (600 MPa for 10 min) and lactate salts showed only a small synergic effect on spoilage bacterial population, with no significant difference in comparison to HPP samples. IV. CONCLUSION

HPP alone was sufficient to ensure a 4 log reduction of Listeria monocytoges strains on frozen beef carpaccio and increase the shelf life of the product. After HPP, potassium lactate and sodium diacetate combination delayed the growth of LAB and psycrothophs during refrigeration storage. Nevertheless, the shelf life extension achieved in this work has been shorter than expected. ACKNOWLEDGEMENTS This work was partially funded by Facultad de Ingeniería y Ciencias Exactas, Universidad Argentina de la Empresa. For this work, Yanina X. Barrio was recipient of a scholarship awarded by Santander Bank. REFERENCES 1. Ethelberg, S., Sørensen, G., Kristensen, B.,

Christensen, K., Krusell, L., Hempel-Jørgensen, A., Perge, A. & NielseNielsen, E. M. (2007). Outbreak with multi-resistant Salmonella Typhimurium DT104 linked to carpaccio, Denmark, 2005. Epidemiology and infection 135 (6): 900–907.

2. Campus, M. (2010). High Pressure Processing of Meat, Meat Products and Seafood. Food Engineering Reviews 2(4): 256–273.

3. Carlez, A., Veciana-Nogues, T., & Cheftel, J.C. (1995). Changes in Colour and Myoglobin of Minced Beef Meat Due to High Pressure Processing. Lebensmittel-Wissenschaft Und- Technologie 28 (5): 528–538.

4. Szerman, N., Barrio, Y., Schroeder, B., Martinez, P., Sancho, A. M., Sanow, C., & Vaudagna, S. R. (2011). Effect of high hydrostatic pressure treatments on physicochemical properties, microbial quality and sensory attributes of beef carpaccio. Procedia Food Science 1: 854–861.

5. Vaudagna, S. R., Gonzalez, C. B., Guignon, B., Aparicio, C., Otero, L., & Sanz, P. D. (2012). The effects of high hydrostatic pressure at subzero temperature on the quality of ready-to-eat cured beef carpaccio. Meat Science, 92(4): 575–581.

6. Garriga, M., Grebol, N., Aymerich, M., Monfort, J. & Hugas, M. (2004). Microbial inactivation after high-pressure processing at 600 MPa in

commercial meat products over its shelf life. Innovative Food Science & Emerging Technologies 5: 451–457.

7. Barmpalia, I.M., Koutsoumanis, K.P., Geornaras, I., Belk, K.E., Scanga, J.A., Kendall, P.A., Smith, G.C. & Sofos, J.N. (2005). Effect of anti-microbials as ingredients of pork bologna for Listeria monocytogenes control during storage at 4º or 10ºC. Food Microbiology 22: 205–211.

8. Fugett, E., Fortes, E., & Nnoka, C. (2006). International Life Sciences Institute North America Listeria monocytogenes Strain Collection: Development of Standard Listeria monocytogenes Strain Sets for Research and Validation Studies. Journal of Food Protection 69 (12): 2929–2938.

9. FSIS (Food Safety and Inspection Service), (2006). Compliance Guidelines to Control Listeria monocytogenes in Post-lethality Exposed Ready-to-Eat Meat and Poultry Products. FDA, USDA. http://www.fsis.usda.gov/oppde/rdad/FRPubs/97013F/LM_Rule_Compliance_Guidelines_May_2006.pdf.

10. Jofre, A., Garriga, M. & Aymerich, T. (2008). Inhibition of Salmonella sp., Listeria monocytogenes and Staphylococcus aureus in cooked ham by combining antimicrobials, high hydrostatic pressure and refrigeration. Meat Science, 78 (1-2): 53–59.

11. Koseki, S., Mizuno, Y., & Yamamoto, K. (2007). Predictive modelling of the recovery of Listeria monocytogenes on sliced cooked ham after high pressure processing. International Journal of Food Microbiology 119 (3): 300–307.

12. Realini, C. E., Guàrdia, M. D., Garriga, M., Pérez-Juan, M., & Arnau, J. (2011). High pressure and freezing temperature effect on quality and microbial inactivation of cured pork carpaccio. Meat Science 88 (3): 6–11.

13. Diez, A, Santos, E., Jaime, I., & Rovira, J. (2009). Effectiveness of combined preservation methods to extend the shelf life of Morcilla de Burgos. Meat Science, 81(1), 171–177.

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ROOIBOS (Aspalathus linearis) IS AN EFFECTIVE FUNCTIONAL INGREDIENT IN RETARDING LIPID OXIDATION OF COOKED RABBIT

MEAT PATTIES

Antonella Dalle Zotte1*, Hannaneh Mousavikhorshidi1,2, Valentino Pizzocchero1,3, Marco Cullere1

1Department of Animal Medicine, Production and Health, University of Padova, Viale dell’Università 16, 35020 Legnaro (PD), Italy 2Department of Agriculture and Eniviromental Science, University of Udine, Via delle Scienze 206, 33100 Udine (UD), Italy

3Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell’Università 10, 35020 Legnaro (PD), Italy *[email protected]

Abstract – In the present trial, the effect of a fermented rooibos tea extract as a natural antioxidant on the shelf-life of rabbit meat patties was investigated. With this purpose, three treatments with the inclusion of 0.5% (R1), 1% (R2) and 2% (R3) fermented rooibos tea extract were compared to a control group without any extract (C: 0%). The inclusion of 0.5%, 1% and 2% fermented rooibos tea extract lowered the peroxides content of rabbit meat patties compared to untreated ones (P<0.0001). Moreover, the rooibos presence affected also the pH values of rabbit meat patties, with R2 and R3 groups providing lower values compared to R1 and C groups (P<0.0001). Differently, although a lower drip loss in patties treated with the rooibos tea extract compared to the control group was expected, no effect was observed in this sense. Consequently, further studies are needed to deeply understand the mechanisms through which the rooibos presence would affect quality aspects of meat and meat products. I. INTRODUCTION Lipid oxidation represents an important issue for the meat industry because it negatively affects meat colour, texture, flavour, and determine loss of nutritional value, ultimately reducing its shelf-life [1]. Synthetic antioxidants like butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) are commonly used to ensure optimal product quality. However, due to safety concerns and a changing consumer’s preference regarding food additives, since the last decade natural antioxidants have been gaining more and more scientific and commercial attention [2]. One emerging natural source of unique phenolic compounds such as aspalathin, is a South

African leguminous shrub named rooibos (Aspalathus linearis) which has been appreciated from centuries by locals as a beverage. Traditional rooibos preparation includes an oxidation (fermentation) step, essential to develop the characteristic sweetish flavour and red-brown colour which determines lower antioxidant levels compared to the unfermented (green) form [3]. The antioxidant activity of phenolic acids and flavonoids found in rooibos tea showed to be comparable to that of α-tocopherol and to the widely used synthetic antioxidants BHT and BHA [4]. Considering food matrixes, the antioxidant potential of rooibos was only tested in two recent studies [5; 6]. Unfermented rooibos was applied to ostrich meat patties in a shelf-life trial, whereas the classic fermented rooibos was evaluated in processed meat products: nitrates and nitrites-free ostrich salami and ostrich droёwors. The preliminary study by [5] provided promising results in terms of lipid stability of fresh ostrich meat and salami. Differently, even if the presence of rooibos generated positive sensory scores, the oxidative stability of the droёwors remained unaffected [6]. Consequently, further confirmations of the positive effects of rooibos on the quality of meat and meat products are required. In the specific case of rabbit meat, which is very rich in unsaturated fatty acids [7], a certain degree of lipid oxidation, mainly during processing, storage and cooking is expected [8; 9]. Thus, in the present trial, we studied the effect of a fermented rooibos extract in prolonging the shelf-life of rabbit patties during display conditions.

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The trial took place at the Department of Animal Medicine, Production and Health of the University of Padova (Italy). A total of 16 kg of rabbit meat from hindleg and loin cuts were ground. The rooibos extract was prepared according to the procedure described by Joubert et al. [10]. Meat was then divided in four batches, and manually mixed with different rooibos inclusion levels: Control (0% rooibos), R1 (0.5% rooibos), R2 (1% rooibos) and R3 (2% rooibos). Then the mixture was ground again in order to allow a better distribution of the rooibos extract. Afterwards, 60 patties per treatment of 50 g each were prepared, individually wrapped with plastic film in order to prevent direct air contact, and then placed in a fridge at 4 °C for a 7 days shelf-life trial. Continuous cold fluorescent light illumination was ensured for the whole period, in order to simulate commercial display conditions. Just before wrapping, n=40 patties per treatment were weighed for drip loss determination and pH was measured twice. The same measurements were repeated at days 1, 3 and 6 of storage. At days 0, 1, 3 and 6 a total of 80 patties (5/treatment/day of storage) were removed from the fridge, freed from the plastic film, wrapped with aluminium sheets and individually identified, vacuum sealed in four plastic bags (one/treatment) and cooked in a water bath for 25 minutes at 80 °C. After cooling, patties were frozen until peroxide value analysis (internal method of the Istituto Zooprofilattico Sperimentale delle Venezie) and expressed as meq O2/kg meat. A One-way Anova [11] tested the rooibos inclusion level as fixed effect and the significance level was calculated at the 5% confidence level. For drip loss determination the initial weight of the patties was considered as covariate. III. RESULTS AND DISCUSSION The presence of rooibos extract tea significantly affected the peroxides content of rabbit meat patties throughout the trial (Table 1), improving their shelf-life. Already at day 0, independently to the inclusion level, patties treated with rooibos tea extract showed a higher oxidative stability compared to the control group (P<0.0001). This

result could depend on the three week frozen storage prior to analysis. As a result, rooibos extract had time to effectively counteract the oxidative process. At days 1 and 3, groups R2 and R3 provided the best protection against lipid oxidation even if also R1 showed a lower peroxides value compared to C group. At day 6, the oxidative degree of the meat worsened in all groups. However, the presence of the rooibos tea extract (R1, R2 and R3) still generated lower peroxides values compared to the untreated samples (C). This result could be interesting for the meat industry in terms of costs, as the inclusion of a 1% rooibos tea extract was enough to provide satisfactory protection against lipid oxidation. Cullere et al. [5] showed a protective effect against oxidation of the same fermented rooibos tea extract on ostrich salami. Specifically, after 15 days of ripening, salami incorporated with 0.5% and 1% rooibos tea extract had a lower TBARS content compared to the control group. Differently, Hoffman et al. [6] did not find any significant effect of fermented rooibos (0.25%, 0.5% and 1% of inclusion level) on ostrich dro wors in terms of protection against lipid oxidation. The only other work that studied the effect of rooibos on fresh meat was that of Cullere et al. [5] where 2% unfermented rooibos was applied to ostrich patties and significantly retarded the oxidative process compared to a control group (0% unfermented rooibos). Table 1. Peroxide (meq O2/kg meat) values (measured at day 0, 1, 3, 6) of cooked rabbit patties without (C) or with 0.5% (R1), 1% (R2) and 2% (R3) fermented rooibos levels Treatments

RSD(1) C R1 R2 R3

No. 20 20 20 20

Day 0 8.4A 3.0B 2.5B 2.4B 0.60

Day 1 10.2A 4.6B 3.3C 2.9C 0.41

Day 3 11.8A 4.4B 3.1C 3.0C 0.45

Day 6 18.3A 6.7B 6.5B 5.8B 1.12 A,B,C Means in the same row with unlike superscripts differ (P<0.0001); (1) Residual Standard Deviation The results presented in Table 2 on the pH values of rabbit meat patties showed that the presence of the rooibos tea extract lowered the pH values compared to the control group (P<0.0001).

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Specifically, increasing the rooibos extract inclusion level, the pH progressively decreased. As a consequence, R3 group provided the best results throughout the experiment followed by groups R2 and R1, with C patties exhibiting always the worst values. These results were probably a consequence of the better oxidative status of the treated patties compared to those belonging to the C group. In the only other research studying rooibos inclusion on fresh meat quality [5] it was observed that the inclusion of a 2% dried unfermented rooibos leaves to ostrich meat patties provided lower pH values compared to a control group until the sixth day of an eight-days shelf-life trial. Table 2. pH values (measured at day 0, 1, 3, 6) for rabbit patties without (C) or with 0.5% (R1), 1% (R2) and 2% (R3) fermented rooibos levels Treatments

RSD(1) C R1 R2 R3

No. 40 40 40 40

Day 0 6.14A 6.10B 6.09B 6.00C 0.02

Day 1 6.26A 6.16B 6.14B 6.07C 0.03

Day 3 6.20A 6.15B 6.13B 6.09C 0.03

Day 6 6.25A 6.23A 6.19B 6.04C 0.02 A,B,C Means in the same row with unlike superscripts differ (P<0.0001); (1) Residual Standard Deviation Results presented in Table 3 dealt with drip loss of rabbit meat patties. The presence of rooibos tea extract had a significant effect on the drip loss between Days 0-1, 1-3 and 3-6. Results at Day 0-1 showed that R2 and R3 groups had higher drip loss values than those exhibited by C and R1 groups (P<0.001). But at Day 1-3 the situation was completely reversed, with a progressive drip loss lowering with increasing rooibos extract inclusion levels. In the third window of time (Day 3-6), the trend of drip loss changed again with C patties showing a lower value compared to R3 group, with R1 and R2 being intermediate (P<0.05). As a result of this inconstant pattern, the total drip loss (Day 0-6) was not significantly affected by treatments (P>0.05). This result was partly unexpected and it was probably the result of the interaction of two different phenomena. On one hand, the protection of the rooibos tea extract against lipid oxidation (Table 1) should have had a positive

effect also on drip loss, as lipid oxidation is known to affect the ability of myofibrillar proteins to entrap water [12]. On the other hand, however, in food products the interaction protein-polyphenol through hydrogen binding is known to occur. The strength of this complex, which depends on the specific structures of the phenolic group of polyphenols and the amide group of protein, would alter charge distribution on the protein surface and consequently also the water holding capacity [13; 14]. Even if there are few studies on the relationship between polyphenols-protein complex and functional properties of proteins, this effect seemed to partly explain our unexpected results and also those of other studies evaluating the ability of natural preservatives to enhance the shelf-life of meat products. In fact, in the above mentioned works, a protection against lipid oxidation didn’t always provide also an improvement in terms of WHC as well as a drip loss lowering [15]. Table 3. Drip losses (%) of rabbit meat patties without (C) or with 0.5% (R1), 1% (R2) and 2% (R3) fermented rooibos levels Treatments

RSD(1) C R1 R2 R3

No. 40 40 40 40

Day 0-1 1.18B 1.06B 2.20A 2.19A 0.46

Day 1-3 2.37A 2.04AB 0.70BC 0.52C 1.16

Day 3-6 0.05b 1.35ab 0.58ab 1.75a 1.44

Day 0-6 3.60 4.45 3.48 4.47 1.19 A,B,C,D Means in the same row with unlike superscripts differ (P<0.001); a,b means in the same row with unlike superscripts differ (P<0.05); (1) Residual Standard Deviation IV. CONCLUSION The practical application of the rooibos tea extract to meat and meat products could generate interesting outcomes for the meat industry. In fact, fermented rooibos tea extract confirmed to be an optimal antioxidant source also for rabbit meat patties enhancing their shelf-life through the reduction of the peroxides content and pH values. However, as literature data on the application of this natural source of antioxidants on fresh meat and meat products is still scarce and results are sometimes contrasting, further studies are needed

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to deeply understand the mechanisms through which the rooibos presence would affect quality aspects of meat and meat products. Moreover, the sensory acceptability of the product should also be further investigated. ACKNOWLEDGEMENTS Authors thank E. Joubert, of Post-Harvest and Wine Technology Division (Agricultural Research Council of South Africa; Infruitec- Nietvoorbij, Stellenbosch, South Africa), for providing the fermented rooibos extract. Research funded by the University of Padova (Ricerca Scientifica fondi quota EX 60% code: 60A08-2910/12). REFERENCES 1. Peña-Ramos, E. A., & Xiong, Y. L. (2003).Whey

and soy protein hydrolysates inhibit lipid oxidation in cooked pork patties. Meat Science 64: 259-263.

2. Andrés, S., Tejido, M. L., Bodas, R., Morán, L., Prieto, N., Blanco, C., & Griáldez, F. J. (2013). Quercetin dietary supplementation of fattening lambs at 0.2% rate reduces discolouration and microbial growth in meat during refrigerated storage. Meat Science 93: 207-212.

3. Joubert, E., & Schulz, H. (2006). Production and quality aspects of rooibos tea and related products. A review. Journal of Applied Botany and Food Quality 80: 138-144.

4. Von Gadow, A., Joubert, E., & Hansmann, C. F. (1997). Comparison of the antioxidant activity of aspalathin with ahat of other plant phenols of rooibos tea (Aspalathus linearis), α-tocopherol, BHT, and BHA. Journal of Agricultural and Food Chemistry 45: 632-63.

5. Cullere, M., Hoffman, L. C., & Dalle Zotte, A. (2013). First evaluation of unfermented and fermented rooibos (Aspalathus linearis) in preventing lipid oxidation in meat products. Meat Science 95: 72-77.

6. Hoffman, L.C., Jones, M., Muller, N., Joubert, E., & Sadie, A. (2014). Lipid and protein stability and sensory evalation of ostrich (Struthio camelus) droёwars with the addition of rooibos tea extract (Aspalathus linearis) as a natural antioxidant. Meat Science 96: 1289-1296.

7. Dalle Zotte, A. (2002). Perception of rabbit meat quality and main major factors influencing the rabbit carcass and meat quality. Livestock Production Science, 75, 11-32

8. Castellini, C., Dal Bosco, A., & Bernardini, M. (1998). Effect of dietary vitamin E on the oxidative

stability of raw and cooked rabbit meat. Meat Science 50: 153-161.

9. Cavani, C., & Petracci, M. (2004). Rabbit meat processing and traceability. Proc. 8th World Rabbit Congress, September 7-10, 2004 – Puebla, Mexico

10. Joubert, E., Viljoen, M., De Beer, D., & Manley, M. (2009). Effect of heat on aspalathin, iso-orientin, and orientin contents and color of fermented rooibos (Aspalathus linearis) iced tea. Journal of Agricultural and Food Chemistry 57: 4204–4211.

11. SAS. (2004). SAS/STAT User’s Guide (Release 9.1) SAS Inst. Inc., Cary NC, USA.

12. Huff-Lonergan, E., & Lonergan, S. M. (2005). Mechanisms of water-holding capacity of meat: the role of post-mortem biochemical and structural changes. Meat Science 71: 194-204.

13. Kawamoto, H., Mizutani, K., & Nakatsubo, F. (1997). Binding nature and denaturation of protein during interaction with galloylglucose. Phytochemistry 46: 473–478.

14. Labieniec, M., & Gabryelak, T. (2005). Measurement of DNA damage and protein oxidation after the incubation of B14 Chinese hamster cells with chosen polyphenols. Toxicology Letters 155: 15–25.

15. Hayes J.E., Stepanyan V., Allen P., O’Grady M.N., Kerry J.P. (2010) Effect of lutein, sesamol, ellagic acid and olive leaf extract on the quality and shelf-life stability of packaged raw minced beef patties. Meat science 84: 613-620.

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EFFECT OF THE INCLUSION OF CHESTNUTS IN THE FINISHING DIET ON THE CHANGES IN CHEMICAL COMPOSITION DURING

THE PROCESSING OF CELTA DRY-CURED HAM

Roberto Bermudez1, Daniel Franco1 , Javier Carballo2, Carril, J.A.3 and José M. Lorenzo1*

1Centro Tecnológico de la Carne de Galicia, Rúa Galicia Nº 4, Parque Tecnológico de Galicia, San Cibrán das Viñas, 32900

Ourense, Spain 2Area de Tecnologia de los Alimentos, Facultad de Ciencias de Ourense, Universidad de Vigo, 32004 Ourense, Spain

3ASOPORCEL, Recinto Ferial El Palomar s/n.27004. Lugo. España * Corresponding author. Tel: +34 988 548 277; fax: +34 988 548 276

E-mail address: [email protected]

Abstract –The effect of the inclusion of chestnut in the finishing diet on chemical composition through the manufacturing process of dry-cured Celta ham was studied. For this work, twenty-four hams of each type (from three different diets: concentrate, mixed and chestnut) were used. During the whole curing period, moisture content decreased progressively and significantly (P<0.001) throughout the process for all groups, reaching average values of 43.36, 45.47 and 44.35 g/100 g for hams from mixed, chestnut and concentrate diets, respectively at the end of process. The intramuscular fat content increased slightly from an initial average value of 8.96 to 12.65 g/100 g of total solids at the end of the “bodega” stage. Significant differences (P<0.01) were only observed among groups after salting and after the first days of drying-ripening. On the other hand, a significant (P<0.001) decrease in the protein content was observed during the manufacturing process for the three dry-cured batches studied, from an initial average value of 86.0 to 75.6 g/100 g of TS at the end of the “bodega” stage. I.- INTRODUCTION Dry-cured ham is a traditional product whose curing process is characteristic of a particular geographical origin and hence, sometimes, regulated by a protected designation of origin (PDO). These protected hams show specific sensory characteristics of aroma, flavour and texture. The use of local breeds as an alternative pig production system has important advantages, as these breeds are closely related to the environment and they help to maintain biodiversity and sustainable agricultural production, especially in depressed areas. However, the rationalisation of production often leads to strong modifications of these traditional systems, with the risk of losing product quality as well as loss of heritage, social and cultural. Galicia (NW Spain) is a great production area of chestnuts (Castanea sativa Mill.). At the present

time, chestnuts are underutilized and this situation contrasts with the high current prices of commercial concentrates for animal feed, so the use of chestnuts in the feeding of the Celta pig breed, in an extensive management system, would allow the reduction of production costs, and putting in the market products of quality, differentiated, and having a high added value and a healthier fat. Information about dry-cured ham from Celta pig is very scarce and only previous studies [1, 2, 3] have been realised on this breed, but no more knowledge is published in the literature. So, the aim of this study was to evaluate the effect of inclusion of chestnut in the finishing diet on the chemical composition throughout the manufacturing process of dry-cured Celta ham. II.- MATERIALS AND METHODS II.1.- Pigs and diets A total of 36 Celta pigs were used in the study. Piglets were vaccinated and deparasitised following the usual protocols. Animals were suckled until an age of 40 days. All pigs were reared and fattened until the age of 16 months in an extensive regime, with a livestock density of 12 animals per hectare. After weaning, the pigs were fed with a concentrate feeding. At the age of 12 months, the pigs were randomly divided into three different groups of 12 animals: Group A was fed concentrate feeding (3 kg/animal and day) for the 4 months prior to slaughter; Group B was fed a mixed diet (concentrate feeding/chestnuts; 1.5 kg concentrate + 2.5 kg chestnuts per animal and day) for the remaining four months, and Group C was fed a mixed diet (concentrate feeding/chestnuts) until the age of 13 months, and then a diet of chestnuts only (5 kg/animal and day) in the three months prior to slaughter. II.2.- Samples

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Seventy two Celta ham pieces of 13.5±1.4 kg were used (twenty-four per treatment). Raw pieces were salted with an excess of coarse salt. A heap was formed alternating layers of ham pieces and layers of salt. In this way, the pieces were totally covered with salt. Hams were salted during 14 days in a salting room maintaining a temperature between 2 and 5 °C and relative humidity between 80 and 90%. After the salting stage, the pieces were taken from the heap, brushed, washed, and transferred to a post-salting room where they stayed for 70 days at 3-6 °C and around 85-90% relative humidity. After the post-salting stage, the pieces were transferred to a room for ripening where, during 240 days, the temperature was moderately raised up to 30 ºC and the relative humidity progressively lowered to 40% to achieve adequate drying. Then, the hams were left to mature for 9 additional months in a chamber under environmental conditions of temperature ranging from 12 to 24 ºC and relative humidity of 70-80%. From each group, samples were taken from raw pieces, after sating, after post-sating, at 80, 160 and 240 days of ripening, and at 90, 170 and 270 days of bodega step. In each sample point two Celta hams of each group were analyzed. II.3.- Chemical composition Moisture, fat, protein (Kjeldahl N x 6.25) and ash were quantified according to the ISO recommended standards [4, 5, 6, 7], respectively. Total chlorides were quantified according to the Carpentier-Vohlard official method [8]. II.4.- Statistical analysis In order to study significant differences among the groups studied, an analysis of variance (ANOVA) and posterior Duncan´s test with a 0.05 level of significance was performed, using the computer programme SPSS package (SPSS 19.0, Chicago, IL, USA). III.- RESULTS AND DISCUSSION The changes in the proximate composition of dry-cured hams from the three finishing diets (mixed, chestnut and concentrate) throughout the manufacturing process are shown in Table 1. The moisture content decreased progressively and significantly (P<0.001) throughout the process for all groups, reaching average values of 43.36, 45.47 and 44.35 g/100 g for hams from mixed, chestnut and concentrate diets, respectively, at the end of process. Significant differences among the pieces that belonged to the different finishing diets were

observed in raw piece (P<0.01), in the first point of drying stage (P<0.05) and in the medium of “bodega” stage (P<0.05). Little differences, however, were observed and in general the trend was similar in the three batches studied. The final mean of moisture contents determined in dry-cured ham was lower than those observed in other dry-cured hams [9, 10]. During the first 14 days of manufacture, there was the most and significant change in the moisture content from the point of view of kinetic’s dehydration, due to the exit of fluids during the salting stage, although there was not significant differences (P>0.05) in this stage among groups. In terms of loss of water, the dehydration was more intense during the drying–ripening stage due to both the duration of this stage and the environmental conditions (higher temperature and lower relative humidity) in the chambers where it takes place. In “bodega” stage, the moisture content become stable in the three groups studied. The intramuscular fat content increased slightly from an initial average value of 8.96 to 12.65 g/100 g of total solids at the end of the “bodega” stage. Significant differences (P<0.01) were only observed among batches after salting and post-salting stage. On the other hand, a significant (P<0.001) decrease in the protein content was observed during the manufacturing process for the three dry-cured batches studied, from an initial average value 86.0 to 75.6 g/100 g of TS at the end of the “bodega” stage. The decrease in the protein content appears to be fundamentally due to the increase in the NaCl content during the salting stage. This decrease was less pronounced in the other stages. Only in the first sampling point of the drying ripening stage significant differences (P<0.01) among groups were observed. The NaCl and ash contents increased significantly (P<0.01) after the salting stage due to the homogeneous distribution of the salt throughout the whole piece. Significant differences were observed among batches (P<0.05) in the drying-ripening stage and “bodega” stage, although the differences were small. At the end of the process, the statistical analysis of the data revealed the absence of significant differences in the NaCl concentration on a dry weight basis among groups. At this point, the average NaCl content was 7.45 g/100 g of TS. This value at the end of the manufacturing process was lesser than the values (9.29-11.4% of TS) reported by other authors for

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dry-cured hams [10, 11]. Other authors reported values even two and three times higher (13–20% of TS) for the salt content in hams [12]. IV. CONCLUSIONS From the obtained results it can be concluded that that the chemical composition of dry-cured

ham from Celta pig is little affected by the inclusion of chestnuts in the finishing diet. However, the utilization “in situ” of this natural resource allows to reduce the feeding costs, and therefore also the final price of this high quality meat product.

Table 1. Evolution of the proximate composition during the manufacture of dry-cured ham from Celta pigs fed with different finishing diets (mixed, chestnut and concentrate)

Raw After salting

After post-

salting

Drying ripening stage Bodega stage

SEM

Sig

Days 0 14 84 164 244 324 414 494 594 Time Moisture (%) Mixed 75.22a7 66.796 63.235 57.61a4 51.913 46.782 44.731 42.72a1 43.361 2.16 *** Chesnut 74.83a7 65.816 62.135 58.12a4 51.283 47.892 43.991 44.85b1 45.471 2.01 *** Concentrate 73.94b7 66.346 62.545 57.00b4 52.213 46.652 44.741 44.72b1 44.351 2.10 *** Sig ** n.s. n.s * n.s n.s n.s * n.s Fat (% DM) Mixed 7.271 9.44a1,2 10.382 10.98a2 11.803 11.303 11.423 12.204 13.215 0.37 ** Chesnut 8.551 15.81b5 11.8323 11.43a2 12.784 11.522 12.744 12.874 12.032 0.40 *** Concentrate 11.081 14.92b 13.553 15.23b4 13.783 12.812 11.891 11.851 12.952 0.32 * Sig n.s. ** n.s ** n.s n.s n.s n.s n.s Protein (% DM) Mixed 87.434 79.363 78.852,3 77.70a2 75.991 77.011,2 75.441 75.401 74.231 0.76 *** Chesnut 85.733 75.331 75.541 76.72a1 75.281 77.381,2 76.001 76.021 76.361 0.68 *** Concentrate 84.873 75.722 75.102 73.59b1 75.042 76.212 76.432 77.022 76.322 0.67 *** Sig n.s. n.s. n.s ** n.s n.s n.s n.s n.s Ash (% DM) Mixed 4.09a1 10.24a2 10.65a2 11.042 11.503 11.203 12.24a4 11.773 11.743 0.46 *** Chesnut 4.01a1 9.03b2 12.55b4 11.353 11.373 11.213 10.70b2 11.132 10.942 0.46 *** Concentrate 3.65b1 9.01b2 11.25a3 11.703 10.753 11.513 12.51a4 11.173 11.153 0.49 *** Sig * * ** n.s n.s n.s ** n.s n.s Chlorides (% DM) Mixed 0.441 7.25a2 7.582 7.04a2 6.732 7.32ab2 7.092 7.36a2 7.422 0.42 *** Chesnut 0.411 6.03b2 7.443 7.34a3 6.762,3 7.68a3 7.663 6.97a2,3 7.503 0.43 *** Concentrate 0.431 6.46b2 7.723 8.02b3 7.423 6.87b2 7.543 8.07b3 7.453 0.44 *** Sig n.s. ** n.s * n.s * n.s * n.s

Significance: *** (P<0.001), ** (P<0.01), * (P<0.05), n.s. (not significant); a-b Means in the same column and parameter not followed by a common subscript letter differ significantly (P<0.05) (differences associated to finishing diet); 1-7 Means in the same row (corresponding to the same parameter) not followed by a common superscript number differ significantly (P<0.05) (differences among sampling points)

ACKNOWLEDGEMENTS Authors are grateful to Xunta de Galicia (The Regional Government) (Project FEADER 2012/64) for the financial support. REFERENCES

1. Bermúdez, R., Carballo, J., Franco, I., Franco, D., García-Fontán, M.C. & Lorenzo J.M. (2011). Efecto de la inclusión de la castaña en las características fisicoquímicas de jamones de cerdo celta. Libro de Actas del VI Congreso Mundial del Jamón, Lugo (Spain).

2. Bermúdez, R., Franco, I., Franco, D., Carballo, J. & Lorenzo J.M. (2012). Influence of inclusion of chestnut in the finishing diet on fatty acid profile of dry-cured ham from Celta pig breed. Meat Science 92:394-399.

3. Lorenzo, J.M., Carballo, J. & Franco, D. (2013). Effect of the inclusion of chestnut in the finishing diet on volatile compounds of dry-cured ham from Celta pig breed. Journal of Integrative Agriculture, 12:2002-2012.

4. ISO (1997). Determination of moisture content, ISO 1442:1997 standard. In International standards meat and meat

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products. Genève, Switzerland: International Organization for Standardization.

5. ISO (1973). Determination of total fat content, ISO 1443:1973 standard. In International standards meat and meat products. Genève, Switzerland: International Organization for Standardization.

6. ISO (1978). Determination of nitrogen content, ISO 937:1978 standard. In International standards meat and meat products. Genève, Switzerland: International Organization for Standardization.

7. ISO (1998). Determination of ash content, ISO 936:1998 standard. In International standards meat and meat products. Genève, Switzerland: International Organization for Standardization.

8. ISO (1996). Determination of chloride content—Part 1: Volhard method, ISO 1841-1:1996 standard. In International standards meat and meat products. Genève, Switzerland: International Organization for Standardization.

9. Armenteros, M., Aristoy, M.C., Barat, J.M. & Toldrá, F. (2011). Biochemical and sensory changes in dry-cured ham salted with partial replacements of NaCl by other chloride salts. Meat Science, 90:361-367.

10. Marušić, N., Petrović, M., Vidaček, S., Petrak, T. & Medić, H. (2011). Characterization of traditional Istrian dry-cured ham by means of physical and chemical analyses and volatile compounds. Meat Science, 88:786-790.

11. Moller, J.K.S., Parolari, G., Gabba, L., Christensen, J. & Skibsted, L.H. (2003). Monitoring chemical changes of dry-cured Parma ham during processing by surface autofluorescence spectroscopy. Journal of Agricultural and Food Chemistry, 51:1224-1230.

12. Gou, P., Guerrero, L., & Arnau, J. (1995). Sex and crossbreed effects on the characteristics of dry-cured ham. Meat Science, 40:21-31.

280


MINIMISING PROTEIN OXIDATION IN RETAIL-PACKED MINCED BEEF USING THREE-GAS MA-PACKAGING

Dimitrios Spanos2, Laura Baussá2, Caroline P. Baron2, Mari Ann Tørngren1*

1 Department of Meat Quality, Danish Meat Research Institute, Taastrup, Denmark 2 National Food Institute, Division for Industrial Food Research, DTU, Kgs. Lyngby, Denmark

*[email protected] Abstract – Minced beef is usually packed in high oxygen modified atmosphere packaging (MAP) with a gas mixture consisting of 70-80% oxygen (O2) and 20-30% carbon dioxide (CO2). Unfortunately, this results in rubbery and less juicy beef patties with a more rancid flavour compared with fresh or non-oxygen packed beef. To establish whether three-gas MAP (O2, CO2 and N2), instead of two-gas MAP (O2 and CO2), would affect sensory attributes, shelf life, protein and lipid oxidation, minced beef was packed in MAP with either 40%, 50% or 80% O2 and 20% or 40% CO2 with N2 as filler gas. When comparing traditional MA-packaging (80% O2 + 20% CO2) with a low oxygen packaging atmosphere (40% O2 + 20% CO2 + 40% N2), the latter is seen to increase the meat oxidative stability during storage but decrease acceptability and shelf life. In contrast, high oxygen MAP (80% and 50% O2) results in more oxidation but a longer shelf life. However, this was not sensorially detectable in the first five days of storage. To maintain shelf life, packaging in 50% O2 + 40% CO2 + 10% N2 or 80% O2 + 20% CO2 is preferable, although this gas mixture will not prevent lipid or protein oxidation in the meat. I. INTRODUCTION Modified atmosphere packaging is widely used in the packaging of fresh and processed foods. Traditionally, red meat is packed in 70-80% oxygen (O2) to obtain an attractive bloom colour and with 20-30% carbon dioxide (CO2) to extend microbiological shelf life (Singh et al. 2011). It has been reported that high oxygen MAP results in less tender and less juicy meat with a more rancid flavour and premature browning (PMB) in beef patties (Clausen 2005). This loss of eating quality is caused by oxidative modifications of lipids and oxidation of structural proteins (Jongberg 2011, Estevez, 2011). However, it is possible to lower the oxygen content in minced beef packaging (50% O2, 30% CO2, 20% N2) and still obtain meat with an acceptable colour and low microbial count as well as a good oxidative stability for up to 14 days at 4°C

(Esmer, 2011). However, the impact on protein oxidation has not been investigated in detail. Therefore, the objective of this study was to investigate the effect of low oxygen three-mixture of gas for MAP on the shelf life, protein oxidation markers and eating quality of minced beef. II. MATERIALS AND METHODS Experiment 1- Raw meat evaluation: Experimental design: Bovine shoulder clod (120 kg, approx. 6% fat) was pre-ground two days after slaughter at a Danish deboning plant using nitrogen for cooling. The meat was then transported to a Danish packing plant, where it was weighed out into 500 g trays and sealed with five different gas mixtures, as presented in Table 1 below. The samples were stored for 13 days at 5°C under fluorescent lighting (1200 lux). Samples were taken out on days 0, 6, 8, 11 and 13 after packaging. Samples for shelf life evaluation were immediately assessed whiles samples for protein and lipid oxidation were vacuum-packed and stored in a freezer at -80°C until analysis. Table 1. Experimental design for experiment 1 - Raw meat evaluation: shelf life, lipid and protein oxidation. 1 x cut Minced 2 days after slaughter 5 x gas 1 2 3 4 5

40% O2

20% CO2

40% N2

40% O2 40% CO2 20% N2

50% O2 20% CO2 30% N2

50% O2 40% CO2 10% N2

80% O2 20% CO2

-

1 x storage

C, for up to 13 days

Shelf life Shelf life was measured based on raw meat odour, colour and overall acceptance of bloomed and degassed meat, 30 minutes after opening of the package, using a 4-point scale, where 1 = no off-odour; 2 = slight off-odour, acceptable; 3 = off-odour, unacceptable; 4 = intense off-odour, unacceptable.

281

Lipid oxidation TBARS determination: Meat samples (10 g) were homogenised for 1 min. in 30 mL of TCA solution (7.5% TCA, 0.1% PG and 0.1% EDTA), using an Ultra-Turrax mixer. The mixture was filtered through Whatman Grade 2 filter paper (Sigma-Aldrich), and 5 mL of 0.002M TBA was added to 5 mL of the filtrate. The solution was then incubated for 40 min. at 100°C. Following incubation, absorbance at 532 nm was measured spectrophotometrically, on a Shimadzu UV-1800 spectrophotometer, against a blank sample and the results were expressed as mg MDA/kg of sample. Protein oxidation Free thiol determination: Meat samples (0.5 g) were homogenised with a Polytron PT1200E system in 10 mL of buffer solution (Trizma base 50 mM, EDTA 1mM, pH 7.4) and 100 μL of freshly prepared BHT solution (1mg/mL in methanol). The homogenate was placed in Eppendorf tubes and centrifuged in a Heraeus Biofuge Pico (Kendro, UK) at 13800 g for 10 minutes. The supernatant was then filtered through a 0.45 mM wheel filter (Sartorius). The free thiol concentration was determined fluorometrically on 50 μL of the filtrate using the Amplite fluorometric Thiol Quantification kit (ATT Bioquest). Fluorescence at Ex/Em of 490/520 nm was measured with a SpectraMAX Gemini fluorometer (Molecular Devices). The results were expressed as percentage compared to the thiol content of initial samples stored in vacuum, representing 100% of the free thiol group content. Statistical analysis: data were analysed using SPSS version 22 (IBM corporation). The model included storage time and gas mixture as fixed effect. Differences between the fixed effects were assessed with the Turkey method p < 0.05. Experiment 2 - Cooked meat evaluation: Experimental design: Non-specified bovine forequarter (800kg, approx. 7.1% fat) was pre-ground five days after slaughter at a Danish deboning plant and standardised to the target fat content of 10-12% under nitrogen cooling. The meat was then transported to an industrial packing plant, where it was minced finely (3 mm particle size) and weighed out into 450 g trays. The trays were then transported to DMRI and MA-packed in the three different gas mixtures

shown in Table 2. Sensory analysis was performed five days after packaging.

Table 2. Experimental design for experiment 2 -Cooked meat evaluation: sensory profiling.

1 x cut Minced beef 6 days after slaughter

3 x gas 1 2 3 40% O2

20% CO2 40% N2

50% O2 40% CO2 10% N2

80% O2 20% CO2

- 1 x storage Sensory analysis: For cooked meat evaluation, the meat in each tray was formed into four 110 g beef patties (with a diameter of approx. 9 cm and a thickness of approx. 1 cm), and cooked on a pre-heated pan (170°C) greased with a thin layer of grape seed oil. The patties were turned every two minutes and cooked to a core temperature of 63°C. Each beef patty was divided between two assessors and served under aluminium covers on pre-heated plates. The cooked beef patties were evaluated by a professional trained sensory panel consisting of eight assessors at the Danish Meat Research Institute. All of the assessors had participated in two training sessions in accordance with ISO 4121, ASTM-MNL 13, DIN 13299 and were trained in sensory assessment of meat. The beef patties were assessed according to descriptors developed during the training of the panel. All beef patties were evaluated on the same day as they were collected from storage on a 15-point unstructured scale anchored at the extremes (0 = low intensity and 15 = high intensity) with 6 replicates per treatment. The descriptive attribute for appearance was: internal colour (doneness). The descriptive attributes for taste were: sweet, acid and bitter. The descriptive attributes for flavour were: beef flavour, warmed-over flavour (WOF), metallic flavour, rancid and stale. The descriptive attributes for texture were: juiciness, cohesiveness, rubbery texture, chewing time and crumbliness. Statistical analysis: data were analysed using Panelcheck V1.4.0. The model included gas mixture as fixed effects, and assessors as random effects. Least squares (LSmeans) were calculated and separated using probability of difference. Levels of

282

significance: p > 0.05 = non-significant (ns), 0.05 > p > 0.01 = *, 0.01 > p >0.001 = **, p < 0.0001 = ***. III. RESULTS AND DISCUSSION Shelf life: The initial psychrotrophic plate count of the raw minced meat, two days after slaughter, was measured at 3.7 log cfu/g. During storage, the appearance and odour of the raw meat were evaluated in relation to acceptability, and, when the average score reached 2.5, it was regarded as predominantly unacceptable. As shown in Table 3, the acceptance limit varies depending on the gas mixture. When using only 40% O2, the meat deteriorates within 7-8 days, while 50% O2 extends the shelf life by 2-4 days. The longest shelf life is achieved for samples packaged in 50% O2 + 40% CO2 + 10% N2 or 80% O2 + 20% CO2.

Table 3. Acceptability limit (approx. storage time for score = 2.5) of

C (n=25). 40% O2 50% O2 80% O2

20% CO2 8 days 10 days 12 days 40% CO2 7 days 11 days -

Lipid oxidation: TBARS revealed that raw meat samples containing 40% O2 were, irrespective of the CO2 content, more stable than meat samples containing 50% or 80% oxygen (Figure 1). Therefore, gas mixtures with 40% oxygen (40/20/40 or 40/40/20) are preferred in relation to oxidative stability. CO2 content did not affect the development of lipid oxidation.

Figure 1. TBARS in raw meat samples from minced

beef stored in different MAP gas mixtures for 6, 8 and 13 days.

Protein oxidation: No impact of the gas mixture was observed on free thiols at six, eight or ten days after packaging. Irrespective of the gas composition, a time-dependent decrease in free thiol groups was observed during storage (Figure 2). By day 11, there was a 30-40% decrease in the fluorescence intensity compared with day 0, irrespective of the gas mixture. Protein oxidation occurred, although it was not clearly associated with the MAP gas composition, in contrast to what has previously been reported (Jongberg, 2011), and this might be linked to the method used in our experiment. Alternatively, the O2 level might need to be lower than 40% to observe any difference between samples.

Figure 2. Percentage of free thiol groups in raw meat samples from minced beef stored in different MAP gas

mixtures for 6, 8 and 11 days. Sensory analysis: Sensory profiling of cooked beef patties packed in 40% O2 + 20% CO2 + 40% N2, 50% O2 + 40% CO2 + 10% N2 or 80% O2 + 20% CO2 after five days of storage showed no statistically significant differences in taste, flavour or texture between the three different gas mixtures (Table 4). If the evaluation had been carried out later in the storage period, attributes related to oxidative changes in texture or flavour might have been significantly different when taking the onset of the oxidative changes into account. Three-gas MAP must be muscle-specific, as reported in an earlier study by Tørngren et al. (2013), which recommended packaging fresh pork chops in three-gas MAP with 40% O2 + 20% CO2 + 40% N2, since eating quality was increased.

0

2

4

6

8

10

12

14

16

18

0 2 4 6 8 10 12 14

mg

MD

A/K

g sa

mpl

e

Storage days

40% O2 / 20% CO2 40% O2 / 40% CO2

50% O2 / 20% CO2 50% O2 / 40% CO2

80% O2 / 20% CO2

0

10

20

30

40

50

60

70

80

90

100

Day 6 Day 8 Day 11

Thi

ol c

onte

nt c

ompa

red

to c

ontr

ol (%

)

40% O2 50% O2 80% O2

283

Table 4. Sensory attributes of cooked beef patties from minced meat stored for 5 days in MAP with different gas mixtures (O2/CO2/N2) at 5°C (n=48).

MAP 40/20/40

MAP 50/40/10

MAP 80/20

p-value

Cooked colour 10.0 10.7 10.4 0.095 Cooked beef 4.8 5.1 5.0 0.396 WOF 3.8 4.3 3.5 0.309 Rancid 3.2 2.2 2.3 0.159 Cohesiveness 5.5 5.0 5.3 0.101 Juiciness 6.2 6.3 6.5 0.232 Rubbery 4.5 4.9 4.7 0.506 Crumbliness 7.2 7.2 6.8 0.209

IV. CONCLUSION

Packaging of minced raw beef in three-gas MAP with 50% O2 + 40% CO2 + 10% N2 will result in the same shelf life as traditional high oxygen MAP with 80% O2 + 20% CO2. In contrast, using a three-gas mixture with 40% O2 + 20-40% CO2 will reduce the shelf life of raw meat at 5°C by 2-4 days compared with 50% O2 + 20-40% CO2.

Reducing the oxygen content in-package

from 50% to 40% O2 delayed lipid oxidation in raw meat. Whereas, protein oxidation was not found to be affected by gas composition, only by storage time.

Protein and lipid oxidation in raw meat remained unaffected when decreasing the carbon dioxide concentration from 40% to 20% CO2.

Sensory profiling of cooked beef patties

showed no benefit related to flavour and texture when using three-gas mixtures with 40-50% O2 instead of a two-gas mixture with 80% O2.

ACKNOWLEDGEMENTS The authors thanks the technicians at the Danish Meat Research Institute (DMRI), Roskilde, Denmark and the Danish cattle levy fund financed this work.

REFERENCES 1. Clausen (2005). Sensory evaluation of ground

beef stored in different atmospheres. 51st International Congress of Meat Science and Technology, August 7-12, Baltimore, Maryland, USA

2. Esmer, O.K., Irkin, R., Degirmencioglu, N., and Degirmencioglu, A. (2011). The effects of modified atmosphere gas composition on microbiological criteria, color and oxidation values of minced beef meat. Meat Science 88:221-226.

3. Estévez, M. (2011). Protein carbonyls in meat systems: A review. Meat Science 89:259-279.

4. Lagerstedt, Å., Lundstrøm, K., & Lindahl, G. (2011). Influence of vacuum or high-oxygen modified atmosphere packaging on quality of beef M. longissimus dorsi steaks after different ageing times. Meat Science 87:101-106.

5. Jongberg, S, Skov, S. H, Tørngren, M.A., Skibsted, L. H. and Lund, M. L. (2011). Effect of white grape extract and modified atmosphere packaging on lipid and protein oxidation in chill stored beef patties. Food Chemistry 218, 276-283.

6. Min, B., Nam, K.C., Cordray, J., and Ahn, D.U. 2008. Endogenous factors affecting oxidative stability of beef loin, pork loin, and chicken breast and thigh meats. Journal of Food Science 73, C439-C446.

7. Singh, P., Wani, A., and Saengerlaub S. (2011). Understanding critical factors for the quality and shelf-life of MAP fresh meat. A review. Critical Reviews in Food Science and Nutrition 51:146-177.

8. Tørngren, M. A., Darré, M. T. & Kristensen, L. (2013). Optimising the eating quality and shelf life of retail pork cuts using muscle-specific three-gas MA-packaging, 59th International Congress of Meat Science and Technology, Izmir, Turkey

284


OPTIMISING EATING QUALITY AND SHELF LIFE OF ENHANCED AND

MARINATED PORK CHOPS USING THREE-GAS MAP

Mari Ann Tørngren1* 1 Department of Meat Quality, Danish Meat Research Institute, Taastrup, Denmark

*[email protected] Abstract – Fresh pork is packed in high oxygen modified atmosphere packaging (MAP) to preserve an attractive bloom colour on the surface of the meat and to extend the microbial shelf life. Unfortunately, the gas composition consisting of 70-80% oxygen (O2) and 20-30% carbon dioxide (CO2) results in less tender and less juicy meat with a more rancid flavour compared with wrapped or vacuum-packed pork. To establish whether three-gas MAP (O2, CO2 and N2) with less oxygen, instead of two-gas MAP, would affect sensory attributes, shelf life and colour stability, enhanced and enhanced + marinated pork chops were packed in MAP with 0%, 40% or 80% O2 and 20% or 40% CO2 an N2 as filler gas. However, it was not possible to optimise the eating quality and shelf life of enhanced pork chops without compromising the acceptance of the raw meat, as seen for fresh pork. By marinating enhanced pork chops, the acceptance limit was extended by two to seven days depending on the gas mixture, and differences in eating quality were equalised. It is recommended to pack enhanced pork chops and enhanced + marinated pork in an anoxic atmosphere (20% CO2 + 80% N2) to preserve the appearance, flavour and texture of the meat. I. INTRODUCTION Modified atmosphere packaging is widely used in the packaging of fresh and processed foods. Traditionally, red meat is packed in 70-80% oxygen (O2) to obtain an attractive bloom colour and in 20-30% carbon dioxide (CO2) to extend shelf life (Singh et al. 2011). Unfortunately, high oxygen MAP results in less tender and less juicy meat with a more rancid flavour and premature browning (PMB) of pork (Lund el al., 2007), beef (Tørngren, 2003, Lagerstedt, 2011, Kim et al. 2010) and chicken (Jongberg 2013). The reduction in eating quality is caused by oxidative changes of lipids and structural proteins (Lund el al., 2007, Jongberg 2011, Estevez, 2011).

The objective of this study was to investigate the effect of low oxygen three-gas MAP on shelf life and eating quality. II. MATERIALS AND METHODS Two retail meat products (1. Enhanced (brine- injected) loin chops, longissimus dorsi (LD), and 2. Enhanced (brine-injected) + marinated loin chops (LD)) were packed in five different gas compositions, stored at 5°C and analysed during storage. For each cut, the experimental work was divided into two sub-trials – part 1: eating quality, and part 2: shelf life. For both sub-trials, all pigs were slaughtered on the same date. However, for Part 1 pigs were selected according to gender (female) and weight (79-87 kg), whereas random pigs were used for Part 2. The same slaughter process was used for both cuts: slaughtering on Monday, pH24, cutting, deboning and injection (10% weight gain) on Tuesday, and slicing and MA-packing on Wednesday (day 0).

Table1. Experimental design

2 x cut Enhanced pork chops (LD)

Enhanced, marinated pork chops (LD)

5 x gas - 20% CO2

80% N2

40% O2 20% CO2

40% N2

40% O2 30% CO2

30% N2

40% O2 40% CO2

20% N2

80% O2 20% CO2

-

Packing: 3 x chops (20 mm) were MA-packed in five different gas compositions (Table 1) with a tray sealer (Multivac, T200, Denmark). Samples for Part 1 were stored for up to nine days at 5°C, and samples for Part 2 were stored for up to 13 days at 5°C. Pigment: hemin (ppm) was measured spectro-photometrically at 640 nm.

285

Psychrotrophic count: samples (total surface) were diluted, surface-plated on PCA and incubated at 6.5°C for ten days. Shelf life: shelf life was measured based on raw meat odour and colour and overall acceptance of bloomed and degassed meat 30 minutes after opening of the package using a 4-point scale, where 1 = no off-odour; 2 = slight off-odour, acceptable; 3 = off-odour, unacceptable; 4 = intense off-odour, unacceptable. Sensory analysis: the meat was tempered at room temperature to 10-12°C and cooked on a frying pan at 170°C to a core temperature of 65-68°C. Samples were evaluated by eight trained assessors using a 15-point unstructured line scale anchored at the extremes (0 = low intensity and 15 = high intensity). The descriptive attributes were developed specifically for each meat product during training, with focus on flavour, texture, juiciness and appearance. Statistical analysis: data were analysed using mixed models (SAS, 9.2, 2002-2008). The model included gas mixture interaction as fixed effects, and assessors, pig and interactions as random effects. Non-significant interactions were deleted from the model. Least squares (LSmeans) were calculated and separated using probability of difference. Levels of significance: p > 0.05 = non-significant (ns), 0.05 > p > 0.01 = *, 0.01 > p >0.001 = **, p < 0.0001 = ***. III. RESULTS AND DISCUSSION Enhanced pork chops Odour is the limiting factor for acceptance of MA-packed enhanced pork chops, regardless of the gas mixture (Table 2), although the meat will deteriorate at different times.

Table 2. Acceptance of enhanced pork (% acceptable samples/days for average score 2.5),

MA-packed in different atmospheres (O2/CO2/N2) stored for 6-7 days at 5°C (n=25).

0/20/80 40/20/40 40/30/30 40/40/20 80/20/0

Day 2 100% 100% 100% 100% 100% Day 5 84% 100% 100% 100% 100% Day 7 0% 32% 84% 68% 92% Day 9 0% 0% 0% 10% 50%

Day 13 0% 0% 0% 0% 4% Colour limit 8 d 8-9 d 8-9 d 11 d 11 d

Odour limit 6 d 6-7 d 7-8 d 7-8 d 8-9 d

Odour limits for traditional MAP in 80% O2 + 20% CO2 and packing in 40% O2 + 30-40% CO2 are comparable. Fewer than 50% of the evaluated samples were unacceptable after approx. eight days of storage. The appearance, texture and flavour of enhanced pork chops are affected by the gas mixture (Table 3). Packing of enhanced pork chops in 30-40% CO2 increases the number of small holes in the cutting surface, although the level is generally very low. Doneness is also affected by the gas mixture, and oxygen seems to be responsible for the changes, with doneness or PMB increasing in the following order: 0% O2 < 40% O2 < 80% O2. Texture is affected with respect to hardness of the first bite, which increases when packing in high oxygen MAP (80% O2 + 20% CO2) or in three-gas MAP with 40% O2 + 30% CO2 + 30% N2 compared with MAP in 0-20% O2 + 20% CO2. Rancid flavour and sour taste are related to the oxygen level in the pack and increase in the following order: 0% O2 < 40% O2 < 80% O2. The significance level between gas 0/20/80 and gas 40/20/40 is as low as p = 0.0619, and between gas 40/40/20 and gas 80/20 p = 0.0645. These samples are therefore considered different.

Table 3. Sensory attributes (1-15) of enhanced pork chops, MA-packed in different atmospheres

(O2/CO2/N2) and stored for 6-7 days at 5°C (n=48)

0/20/8

0 40/20/

40 40/30/

30 40/40/

20 80/20/

0 p

Small holes

0.5a 0.6a 1.1b 0.9b 0.6a 0.0017

Doneness 7.0a 8.5b 8.7b 8.8bc 9.4 c < 0.0001

Tenderness 8.3 8.2 7.6 7.8 7.5 Ns

Hardness 5.0 a 5.0 a 6.1 c 5.3ab 5.8bc 0.0061

Juiciness 7.9 7.6 7.2 6.9 7.6 Ns

Stale 2.9 4.0 4.1 4.1 4.5 Ns

Rancid 1.0a 1.9ab 1.7a 1.8ab 2.6b 0.0263

Pork 5.3 4.7 4.6 4.5 4.5 Ns

286

Sour 1.7a 2.6a 2.5a 2.4a 3.6b 0.0042

In comparison with traditional MAP, two-gas MAP with an anoxic atmosphere (20% CO2 + 80% N2) optimises the appearance, texture and flavour of MA-packed enhanced pork (Table 3). Packing in three-gas MAP (40% O2 + 40% CO2 + 20% N2) optimises appearance and flavour, but to a smaller degree. In an earlier study by Tørngren et al. (2013), packing in three-gas MAP with 40% O2 + 20% CO2 + 40% N2 was recommended for fresh pork chops. For enhanced pork, 40% CO2 is needed to achieve an acceptable shelf life. Enhanced + marinated pork chops Odour is the limiting factor for acceptance of MA-packed enhanced + marinated pork chops (Table 4). In general, the marinade keeps the odour and the colour acceptable for longer compared with enhanced pork chops (Table 2), and the meat is perceived as acceptable for at least 13 days when packed in an anoxic atmosphere (20% CO2 + 80% N2).

Table 4. Acceptance of enhanced + marinated pork (% acceptable samples/days for average

score 2.5), MA-packed in different atmospheres (O2/CO2/N2) and stored for 6-7 days at 5°C

(n=25).

0/20/80 40/20/40 40/30/30 40/40/20 80/20/0

Day 2 100% 100% 100% 100% 100% Day 5 100% 100% 100% 100% 100% Day 7 100% 96% 100% 92% 100% Day 9 72% 64% 80% 48% 92% Day 13 100% 28% 52% 32% 56% Colour limit

>13 d >13 d >13 d >13 d >13 d

Odour limit

>13 d 11 d 13 d 9 d 13 d

In contrast to enhanced pork (Table 3), only the appearance and flavour are affected by the MAP gas mixture, whereas texture remains unaffected when the marinade is added to the surface (Table 5). With respect to appearance, the number of small holes increases when the gas mixture contains 40% CO2 compared with 20% or 30% CO2. Premature browning (PMB), measured by the

attribute doneness, is reduced only when packing the meat in an anoxic atmosphere (20% CO2 + 80% N2). Furthermore, the anoxic atmosphere decreases the stale flavour and increases the pork flavour of the meat. The high level of acceptance of the enhanced + marinated pork chops could partly be due to the masking of the off-flavour and discolouration by the red and flavourful marinade. Nevertheless, colourful marinades can be a way to enhance consumer acceptance of meat packed in an anoxic atmosphere, as the red surface might look more attractive than the purple deoxymyglobin. Table 5. Sensory attributes (1-15) of enhanced + marinated pork chops, MA-packed in different

atmospheres (O2/CO2/N2) and stored for 6-7 days at 5°C (n=48).

0/20/8

0 40/20/

40 40/30/

30 40/40/

20 80/20/

0 p

Small holes 0.6 a 0.7 a 0.7 a 1.1 b 0.6 a 0.0202 Doneness 5.9a 7.4b 8.3b 8.3b 8.1 b 0.0002 Tenderness 9.0 8.3 8.3 8.1 8.7 Ns Hardness 4.7 5.3 5.2 5.6 5.3 NS Juiciness 7.7 7.7 7.6 8.1 8.2 Ns Stale 1.8a 3.6b 3.6b 2.8ab 3.0b 0.0136 Rancid 0.4 1.1 1.2 0.7 0.9 Ns Fried pork 3.0 b 2.5 a 2.5 a 2.8 ab 2.6 a 0.0357 Sour 1.2 2.5 2.3 2.1 2.4 Ns IV. CONCLUSION Enhanced pork should be MA-packed in a two-gas anoxic atmosphere (20% CO2 + 80% N2) to optimise the appearance, flavour and texture of the meat. Unfortunately, it will shorten the shelf life by approx. two days at 5°C. To maintain a shelf life comparable to high oxygen MAP, it is recommended to pack in a three-gas MAP (40% O2 + 40% CO2 + 20% N2), although this gas mixture will optimise only appearance and flavour, and to a smaller degree. It is recommended to MA-pack enhanced + marinated pork chops in an anoxic atmosphere (20% CO2 + 80% N2), since shelf life remains uncompromised, and appearance and flavour are optimised.

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ACKNOWLEDGEMENTS The author thanks the technicians at the Danish Meat Research Institute (DMRI), Roskilde, Denmark. This work was financed by the Danish Pig Levy Fund, Nortura and HK Scan. REFERENCES 1. Estévez, M. (2011). Protein carbonyls in meat

systems: A review. Meat Science 89:259-279. 2. Lagerstedt, Å., Lundstrøm, K., & Lindahl, G.

(2011). Influence of vacuum or high-oxygen modified atmosphere packaging on quality of beef M. longissimus dorsi steaks after different ageing times. Meat Science 87:101-106.

3. Lund, M. N., Lametsch, R., Hviid, M. S., Jensen, O. N., and Skibsted, L. H. (2007). High-oxygen packaging atmosphere influences protein oxidation and tenderness of porcine longissimus dorsi during chill storage. Meat Science 77:295-303.

4. Jongberg, S, Skov, S. H, Tørngren, M.A., Skibsted, L. H. and Lund, M. L. (2011). Effect of white grape extract and modified atmosphere packaging on lipid and protein oxidation in chill stored beef patties. Food Chemistry 218, 276-283.

5. Jongberg et al (2014). Effect of high-oxygen atmosphere packaging on oxidative stability and sensory quality of two chicken muscles during chill storage. Food Packaging and Shelf Life 1, 39-48.

6. Kim,Y.H. Huff-Lonergan,E., Sebranek, J.G., and Lonergan, S.M. 2010. High-oxygen modified atmosphere packaging system induces lipid and myoglobin oxidation and protein polymerization. Meat Science 85:759-767.

7. Singh, P., Wani, A., and Saengerlaub S. (2011). Understanding critical factors for the quality and shelf-life of MAP fresh meat. A review. Critical Reviews in Food Science and Nutrition 51:146-177.

8. Tørngren, M. A. (2003). Effect of packing method on colour and eating quality of beef loin steaks. 49th International 49th International Congress of Meat Science and Technology. Brazil, September. 495-496.

9. Tørngren, M. A., Darré, M. T. & Kristensen, L. (2013). Optimising the eating quality and shelf life of retail pork cuts using muscle-specific three-gas MA-packaging, 59th International Congress of Meat Science and Technology, Izmir, Turkey.

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INFLUENCE OF FREEZING TIME ON THE WATER ACTIVITY AND LIPID OXIDATION IN MEAT OF BROILERS REARED AT THE

CONVENTIONAL AND ANTIBIOTIC-FREE SYSTEMS

Aline Giampietro-Ganeco1, Hirasilva Borba2, Juliana L. M. Mello1, Ana Paula de O.

Saccomani3, Rodrigo A. de Souza3 and Pedro A. de Souza2

1Post-graduation students at São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil

2Professor of Technology Department at São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil

3Post-graduation students at Instituto de Zootecnia - APTA/SAA, Nova Odessa, São Paulo, Brazil

*[email protected]

Abstract – The aim of this study was to evaluate the water activity and lipid oxidation in frozen breast meat from broilers reared at the conventional and antibiotic-free creation systems. Carcasses from 120 broilers of Cobb lineage, from conventional and antibiotic-free creation systems, were used in this study. Were analyzed the water activity and lipid oxidation (TBARs) in fresh breast meat and after three and six months under freezing. The water activity values decreased in broiler’s meat from antibiotic-free system. The meat from conventional broilers showed higher lipid oxidation during storage period. The storage influences the water activity and lipid oxidation values in breast meat from broilers from antibiotic-free and conventional creation systems. I. INTRODUCTION Classified as a healthy food, the chicken meat is used for food, by presenting rich protein content of good quality and is recommended, its consumption, the people of all age groups. But the great majority of animal foods can deteriorate easily. Therefore one way to preserve these foods is the system from freezing. Frozen foods have an enormous growth in the market. Freezing is one of the best methods of food preservation. The process of freezing appropriate limits microbial growth, reduces the speed of chemical reactions, such as enzyme activity and lipid oxidation. At the same time, the freezing process retains the flavor, aroma, color and nutritional value. The freezing process is to reduce the temperature of the food, crystallization from one part of water and some solutes. The benefits are the strengthening of structures and removal of free water from the food. Therefore the importance of studying the activity of water (aw) frozen foods. The deterioration of the meat due to fat oxidation is a factor limiting the shelf life of meat and frozen meat also. The meat from poultries and swine oxidizes more rapidly than

bovine meat. Lipid oxidation is the principal process by which the loss of quality of meat and its products. The aim of this study was to evaluate the water activity and lipid oxidation in frozen breast meat from broilers reared at the conventional and antibiotic-free creation systems. II. MATERIALS AND METHODS Carcasses from 120 broilers of Cobb lineage from antibiotic-free and conventional creation systems, with 42 days of age, purchased from a commercial slaughterhouse were used in this study. Later, the carcasses were sent to Technology Laboratory of Animal Products in São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil, maintaining the cold chain, for meat tenderness tests. Forty carcasses were considered refrigerated carcasses. The other 80 carcasses were brought to the freezing tunnel (fast freezing), and stored for up to three (40) and six months (40) under freezing. Were analyzed the water activity (aw) and lipid oxidation (TBARs) in fresh breast meat (Pectoralis major muscle) and after three and six months under freezing. The aw was measured using the water activity analyzer Aqualab (Decagon Devices Inc.), that bases its measure on the dew point technique. The lipid oxidation was evaluated by measuring substances reactive to thiobarbituric acid (TBARS) according Pikul et al. [1]. For statistical analysis a completely randomized design in 2x3 factorial was used with two creation systems (antibiotic-free and conventional systems) and three freezing periods (0, 3 and 6 months), in forty replications. Data were submitted to analysis of variance using the

289


GLM Procedure and means compared by Tukey test (5%) using the statistical program SAS [2]. III. RESULTS AND DISCUSSION Table 1 shows the average results obtained from water activity (aw) and TBARs analysis in breast meat from broilers from antibiotic-free and conventional creations systems.

Table 1 Water activity (Aw) and test substances reactive to thiobarbituric (TBARs) of breast meat

aw TBARs (mg TMP/kg)

Creation Systems Antibiotic Free 0.983 A 0.413 B Convencional 0.980 A 0.893 A P-value 0.2224 <0.0001

Freezing periods Refrigerated 0.990 A 0.079 C 3 months frozen storage

0.980 B 0.545 B

6 months frozen storage

0.974 C 1.334 A

P-value 0.007 <0.0001 CV(%) 0.89 6.32

Averages followed by different letters differ according to Tukey’s test. The following abbreviations are used: CV: Coefficient of variation

There was no statistical difference between creation systems to analysis that evaluates to water activity values. There was no difference to water activity with the storage period and are shown in Table 2. In broilers meat from the creation of antibiotic-free system decreased the values of water activity. There was a decrease of water activity values in meat from antibiotic-free broilers during the storage process.

Table 2 Breakdown of the interaction between creations systems and period of storage for water

activity

Refrigerated 3 months frozen storage


Antibiotic Free 0.992 Aa 0.985Aab 0.971 Ac Convencional 0.987 Aa 0.976 Aa 0.977 Aa

For a given factor, means followed by different capital letters in the column and different lowercase letters in the row are differ according to Tukey’s test. For lipid oxidation analysis, were observed significant difference between creation systems and storage periods, as shown in the Table 3.

Breast meat from broilers from conventional system showed higher lipid oxidation during storage period than broilers from antibiotic-free system. Carcasses only refrigerated showed lower TBARS values, indicating that the freezing don't interrupts the lipid oxidation.

Table 3 Breakdown of the interaction between creations systems and period of storage for test substances reactive to thiobarbituric (TBARs)



Antibiotic Free 0.054 Bc 0.345 Bb 0.939 Ba Convencional 0.105 Ac 0.745 Ab 1.829 Aa

For a given factor, means followed by different capital letters in the column and different lowercase letters in the row are differ according to Tukey’s test.

Lipid oxidation is a spontaneous and inevitable phenomenon, and the storage of meat under freezing not can to interrupt this process. The meat from "antibiotic-free broilers" showed lower TBARS values, it is possible that these animals receive food with some source of natural antioxidant that contributes to this lower lipid oxidation. IV. CONCLUSION

The storage under freezing influences the water activity and lipid oxidation of breast meat from broilers from antibiotic-free and conventional creation systems. ACKNOWLEDGEMENTS This study was financially supported by Foundation for Research Support in the State of São Paulo (Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP, 2012/10276-0) and supported by Korin Agriculture to whom the authors want to express their gratitude. REFERENCES

1. Pikul, J. (1989). Evaluation of three modified TBA methods for measuring lipid oxidation in chicken meat. Journal of Agriculture and Food Chemistry. 37:1309-1313.

2. Sas Institute. (2002). SAS user’s guide: statistics. Release 8.02. Cary.

290


DETERMINATION OF COOKING WEIGHT LOSS IN DIFFERENT TYPES OF BROILERS AND FREEZING PERIODS

Aline Giampietro-Ganeco1, Hirasilva Borba2, Juliana L. M. Mello1, Luiz C. M. Junior3, Ana

Beatriz B. Rodrigues1, Caroline Demarchi3 and Leonardo D. do C. Vieira 1

1Post-graduation students at São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil

2Professor of Technology Department at São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil

3Graduation students at São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil

*[email protected]

Abstract – The aim of this study was to evaluate cooking weight loss of broilers breast meat from the different types of creations systems and different times storage under freezing. Were analyzed the cooking weight loss in fresh breast meat (Pectoralis major muscle) and after three and six months under freezing. The statistical analysis a completely randomized design in 4x3 factorial was used with four creation systems (Colonial, conventional, antibiotic-free and organic systems) and three freezing periods (0, 3 and 6 months), in forty replications. There was statistical difference between creation systems and freezing periods to analysis that evaluates to cooking weight loss. Those meats were stored for 6 months under freezing showed higher values for weight loss in cooking, The process of freezing food can physically affect the meat. This study conclude different creation systems and storage under freezing influences the cooking weight loss of breast meat from broilers. I. INTRODUCTION Currently, the production of broilers adopts important criteria of productivity, such as carcass yield, breast meat yield and legs and carcass quality and meat. The degree of importance of these features varies according to the company, the type of product sold and the market to which it is intended. Improvement technological meat quality has become an important issue for the poultry industry, since it must offer products well suited to processing and responding to consumer demand. The genetic and the relationship between attributes of meat and other features of interest in broilers features can support an accurate and appropriate strategy to market the property. Consumers are shown to be choosy about the quality of food, they are also more concerned about the impact of agricultural production on the environment and social well-being of the population. In this way, alternative systems of production of broilers, as the rustic, natural and organic seeking to meet a portion of consumers

who prioritize the consumption of healthier products considered were developed. To meet the demands of the markets the quality of broiler meat depends on factors such as cooling rate and temperature. The demand of this product depends on the quality perception of the consumer customer. Are considered as objective criteria for its good acceptance items: appearance, water retention capacity, juiciness, tenderness, skin color, flesh color and cooking weight loss [1]. The aim of this study was to evaluate cooking weight loss of broilers breast meat from the different types of creations systems and different times storage under freezing. II. MATERIALS AND METHODS Carcasses from 120 broilers of colonial, 120 broilers of conventional, 120 broilers of antibiotic-free, and 120 broilers of organic creation systems, purchased from a commercial slaughterhouse were used in this study. The poultry must have access to paddocks and they should have at least three square meters available for each bird housed. The minimum age is 85 days, the use of specific strains for this type of farming is needed. It is the system used on poultry farms for commercial exploitation, with genetically selected for high growth rate lines and excellent feed efficiency, anticoccidial, growth promoters, chemotherapy and ingredients of animal origin and the birds are slaughtered at an average of 42 days. The antibiotic-free broilers are raised in a system without the use of antibiotics, anticoccidial, performance enhancers based antibiotic, chemotherapy and ingredients of animal origin in the diet. This production system has no restriction lines. Are raised in closed sheds and controlled environment and slaughtered at an average of 45 days.

291


The poultry are raised in an organic system presents the pasture area with low density and feed containing ingredients certified organic vegetables. Apart from the chemotherapeutic products should not be used in creating and slaughtered at an average of 45 days. Forty carcasses of each creation system were considered refrigerated carcasses. The other 80 carcasses of each creation system were brought to the freezing tunnel (fast freezing), and stored for up to three (40) and six months (40) under freezing. The carcasses with three and six months were stored in plastic bags traditional under a storage freezing (-18 ° C). Later, the carcasses were sent to Technology Laboratory of Animal Products in São Paulo State University – UNESP, Jaboticabal, São Paulo, Brazil, maintaining the cold chain, for meat cooking weight loss. Were analyzed the cooking weight loss in fresh breast meat (Pectoralis major muscle) and after three and six months under freezing. The cooking weight loss was determined by cooking the samples in a water bath according to methodology described by Honikel [2]. For statistical analysis a completely randomized design in 4x3 factorial was used with four creation systems (Colonial, conventional, antibiotic-free and organic systems) and three freezing periods (0, 3 and 6 months), in forty replications. Data were submitted to analysis of variance using the GLM Procedure and means compared by Tukey test (5%) using the statistical program SAS [2]. III. RESULTS AND DISCUSSION Table 1 shows the average results obtained from cooking weight loss analysis in breast meat from broilers from colonial, conventional, antibiotic-free and organic creation systems. And there was difference to creation systems with the freezing periods and are shown in Table 2.

Table 1 Cooking weight loss of breast meat from different creation systems under freezing periods

Cooking Weight Loss (CWL) Creation Systems

Colonial 27.49 C Convencional 29.14 D Antibiotic Free 25.91 B Orgânico 30.54 A P-value <0.0001

Freezing periods Refrigerated 26.34 C 3 months frozen storage 28.23 B

6 months frozen storage 30.44 A P-value <0.0001 P-value 0.0002 CV(%) 6.35

Averages followed by different letters differ according to Tukey’s test. The following abbreviations are used: CV: Coefficient of variation

There was statistical difference between creation systems and freezing periods to analysis that evaluates to cooking weight loss. Those meats were stored for 6 months under freezing showed higher values for weight loss in cooking, The process of freezing food can physically affect the meat. The freezing is carried out through rapid process, which avoids the formation of large crystal ice in the meat. The formation of ice crystals causes the disruption of cellular structures by drilling, releasing the liquid in the cellular thawing and cooking meat process, visualized in this study.

Table 2 Breakdown of the interaction between creations systems and freezing periods for cooking

weight loss



Antibiotic Free 23.06 Bb 25.08 Bb 29.88 Ba Colonial 26.92 Aa 27.25 Ba 28.14 Ba Conventional 27.08 Ab 30.21 Aa 30.32 Ba Organic 28.30 Ac 30.40 Ab 33.41 Aa

For a given factor, means followed by different capital letters in the column and different lowercase letters in the row are differ according to Tukey’s test. The meat from the organic rearing system showed higher cooking weight loss during all months of storage. The cooking weight loss is an important quality measure as associated with this meat yields after preparation for consumption. The meats of colonial systems didn´t differ with storage for 6 months in a freezing. IV. CONCLUSION

The different creation systems and storage under freezing influences the cooking weight loss of breast meat from broilers. ACKNOWLEDGEMENTS This study was financially supported by Foundation for Research Support in the State of São Paulo (Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP, 2012/10276-0) and supported by

292


Korin Agriculture to whom the authors want to express their gratitude. REFERENCES

1. BUENO, L. G.F. Diagnóstico do Uso de um Frigorífico de Frangos de Corte enfatizando Medidas de Eficiência Energética. Universidade Estadual de Campinas. 2008. Disponível em http://libdigi.unicamp.br/document/?code=vtls000444266. Acesso em 26 de março de 2009.

2. Honikel, K. O. (1987). The water binding of meat. Fleischwirttsch. 67:1098-1102.

3. Sas Institute. (2002). SAS user’s guide: statistics. Release 8.02. Cary.

293


EFFECTS OF PORK ROUND POWDER ADDITION ON THE QUALITY CHARACTERISTICS OF NOODLE PRODUCT

Kihong Jeon1, Youngboong Kim, Youngho Kim, Yoonseon Hwang and Jinyoung Choi2

1Division of Convergence Technology, Korea Food Research Institute, Sungnam, Korea

2 Division of Food Science and Culinary Arts, Shinhan University, Euijeongbu, Korea

*[email protected]

Abstract – The quality characteristics like color, viscosity, texture profile, tension, cooking properties and sensory evaluation were tested to find out the effect of pork round powder treatments in the noodle at the ratio of 1%, 3%, 5% and 7% respectively. In the color test, with the increment of pork round powder ratio, L value and b value decreased but a value increased. In the test of viscosity, the result of peak (P), trough (H), breakdown (P-H), final viscosity (C) and setback (C-P) was found highest in 1% treatment and it decreased with the pork round powder ratio increased. With this experiment, 3% of pork round powder treatment was the best condition for noodle manufacturing because it was most similar condition with the control (p<0.05). In the test of texture profile, the higher ratio of pork round powder in the noodle, its hardness was getting higher while tensile force and tensile distance were down. Tensile force in the 3% treatment was 44.15g which showed similar result with the control of 44.65g (p<0.05). In the turbidity test of the noodle soup, it went up while the pork round powder ratio was high, which resulted that solid in the noodle moved to the soup. Sensory evaluation of color, flavor and taste test resulted also reasonable in the 1% and 3% treatment (p<0.05) and there was no significant difference in the 5% treatment with control in the overall acceptability. Key Words – Pork round powder, Noodle, Viscosity, Texture, Sensory evaluation, I. INTRODUCTION Meat consumption pattern in Korea is normally for roasting uses. Pork belly part is very popular in Korea because it is believed the best part for roasting with its rich flavor comes from fat. This results pork belly part should be imported for short of domestic supply. In the other hands, the round part is called ‘less preferred part’ in the market because it is not good for the roasting with less fat contents and results oversupplied because of its less consumption. With this unique situation of unbalanced meat consumption in Korea, many researches are trying to find the

solution to make up demand and supply balance with development of popular meat processed products including pork round part. Noodle is very common food around the world for a long time. It is made mainly by wheat flour, salt and water after mixing, kneading dough and cutting in a certain form [1]. Nowadays, many convenient types of noodle products are found in the market and the consumption of those kinds of noodle products is increasing. But they seem to focus for its convenient purposes not much for its nutrition value. Now we believed that new concept noodle products with higher nutrition value but maintain its convenience function and texture profile at the same time need to be developed. In this study, new technology for binding capacity between wheat flour and meat powder was tried and physico-chemical quality of noodle with pork round powder was researched. II. MATERIALS AND METHODS Samples Treatments

Material like wheat flour, pork round powder, salt, gluten and water were mixed with the ratio in Table 1 and aged for 30 min. Mixed material of paste was passed through pasta roller (KitchenAid, K5SS, USA) several times until it had same thickness of 2 mm and then cut at same thickness with pasta cutter. Each noodle was cooked at 100℃ for 3 min in separated pots and washed with cold water for 30 sec. Pork round powder was made with chopping (Hand Blender, MR5550MCA, Braun, Spain), deep freezing (VIP Series, Sanyo, Japan), freeze drying for 48 hrs in a freeze drier (Ilshin FD5512, Ilshin, Korea) and then sieving at 20 mesh (Test sieve, 850 ㎛, Chuggye, Korea) before experiment. In this experiment, 3 samples of noodle were prepared and analyzed with triple replicates.

294


Table 1 Mixture formula of noodle with various ratio of pork round powder addition

(Unit: %)

PP-1,3,5,7% : Pork Round Powder 1,3,5 and 7% addition

Analysis Color : L value (lightness), a value (redness) and b valve (yellowness) were detected by color difference meter (Chroma meter, CR-410, Minolta Co., Japan) at the condition of L: 97.10, a: -0.17 and b: 2.08 in the standard white plate. Viscosity : Rapid Visco Analyzer (RVA, New Port Scientific Pty, Ltd, Australia) was used to evaluate the viscosity with the method of AACC [2] and resulted with value of Rapid Visco Unit (RVU). Texture Profile Analyzer (TPA) : Texture was experimented by texture analyzer (TA-XT, stable micro system. UK) with a p20∼40 mm round probe under the condition of pre-test speed 0.5 mm/s, test speed 0.5 mm/s, post speed 0.5 mm/s, strain 50% and 75% distance. Tension : Tension was tested by texture analyzer (TA-XT2i, stable micro system. UK) under the condition of pre-test speed 5.0 mm/s, test speed 5.0 mm/s and post speed 5.0 mm/s. Force (g) and distance (mm) of the cooked noodle were extended until it cut after gripping the end of noodle hold tight. Water Absorption ratio (WAR) : WAR of the noodle was test with below calculation method. WAR (%)=Noodle wt (after cooking-before cooking)

/Noodle wt before cooking x 100 Volume : Increased volume of the cooked noodle was measured with mess cylinder. Turbidity : Turbidity of the cooked noodle soup was tested by the spectrophotometer (Mecasys, Optizen 2120UV, Korea) at 675 nm of Optical Density (OD). Sensory evaluation : Trained 20 panel evaluated the cooked noodle with 9-point scale. Statistics Analysis : ANOVA test was done with SAS/PC+ program [3]. Significance test of the differences between samples was done with Duncan’s multiple range test (p<0.05).

III. RESULTS AND DISCUSSION Color Color of cooked noodles with the various mixture ratio of pork round powder (PP) 1∼7% is resulted in Table 2. Lightness (L) value of control was 70.79 but decreased to 60.36 as the PP ratio increased up to 7% treatment. Redness (a) value of control was -1.82 but increased while the PP ratio was getting higher up to 1.95 at the 7% treatment. Yellowness (b) value was also went down from 15.30 in the control to 11.54 in the 7% treatment (p<0.05). The changes of color was affected by the mixture ratio of pork round powder and also considered to influence to the palatability. The papers which studied the color changes of noodles with the additives like shiitake mushroom (Lentinula edodes) [4], Sanghwang mushroom (Phelllinus linteus) [5], king oyster mushroom (Pleurotus eryngii) [6] or kudzu powder [7] also showed the same result L value decreased at the higher mixture ratio but increased a value and b value in the same conditions.

Table 2 Color changes of cooked noodle with various ratio of pork round powder addition


a-e Mean with same letter in each column are not significantly different by Ducan’s multiple range test (p<0.05).

Viscosity Viscosity of the various mixture ratio of pork round powder (PP) 1∼7% is resulted in Table 3. With the higher ratio of PP, peak viscosity (P), trough (H), breakdown (P-H), final viscosity (C), setback (C-P) of the powder mixture declined gradually except 1% of PP treatment. In the peak viscosity, 72.89 RVU was in the control while 75.55 RVU in 1% treatment and 71.22 RVU in 3% treatment respectively, which showed no significant difference with the pork round powder mixture ratio. But in the 5% and 7% treatment, peak viscosity decreased 65.75 RVU and 61.42

Flour PP Water Salt Gluten

Control 71 0 27 1 1

PP-1% 70 1 27 1 1

PP-3% 68 3 27 1 1

PP-5% 66 5 27 1 1

PP-7% 64 7 27 1 1

Treat- ment

Hunter’s color value L a b

Control 70.79±0.30a -1.82±0.04d 15.30±0.06a PP-1% 69.64±0.15b -1.25±0.01c 14.53±0.18b PP-3% 67.12±0.44c -0.55±0.04b 14.15±0.13c PP-5% 60.94±0.16d 1.95±0.00a 11.88±0.04d PP-7% 60.36±0.29d 1.95±0.02a 11.54±0.13e

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RVU each, this result is believed that meat powder which has no amylose related more to the viscosity than the other treatments. When peak viscosity is high, the texture of noodle is getting strong but the noodle becomes too soft when it is cooked [8]. Trough (H) had highest result of 56.70 RVU in 1% treatment and 51.44 RVU in 3% treatment but there was no significant difference compare to the 53.33 RVU in the control. But it went down to 48.50 RVU and 44.67 RVU in the 5% and 7% treatment respectively. Breakdown (P-H) and final viscosity (C) has also no significant differences between control and 1% and 3% treatment. Setback (C-P) is the difference value of trough deducted from final viscosity which can predict the retrogradation tendency. With the result, all the treatments except 1% treatment shows slower retrogradation than the control [9].

Table 3 Viscosity changes of cooked noodle with various ratio of pork round powder addition

(Unit: RVU)


a-c Mean with same letter in each column are not significantly different by Ducan’s multiple range test (p<0.05).

Texture profile The result of texture profile analysis of cooked noodle made with various ratio of pork round powder in the mixture is in Table 4. Hardness in the control was 1,123.97g and it grew up when the additional ratio of pork round powder increased. In the 7% treatment, it resulted highest value of 1,448.33g among the treatments (p<0.05). There were no significant differences in the springiness and cohesiveness between control and treatments while gumminess and chewiness increased when the mixture ratio of pork round powder was high (p<0.05).

Table 4 Texture profile changes of cooked noodle with various ratio of pork round powder addition


a-c Mean with same letter in each column are not significantly different by Ducan’s multiple range test (p<0.05).

Tension In the result of test on tensile force and tensile distance of cooked noodle with various mixture ratio of pork round powder is shown in the Fig.1. The tensile force of noodle was 44.65g in the control but it got down gradually to 33.54g in the 7% treatment (p<0.05). Tensile distance was 110.68mm in the control but decreased to 81.75mm in 7% treatment which showed also it became shorter when the mixture ratio increased. But 3% treatment had no significant differences compare to control. And it results the best condition in the treatments.

Fig. 1. Tensile force and tensile distance changes of

cooked noodle with various ratio of pork round powder addition


a-d Mean with same letter in each column are not significantly different by Ducan’s multiple range test (p<0.05).

Cooking properties The result of water absorption ratio, volume and soup turbidity of the cooked noodle with various mixture ratio of pork round powder is shown in the Table 5. Water absorption ratio of the cooked

P H P-H C C-P

Con-trol

72.89±11.09a

53.33±8.53ab

19.56±8.53a

109.97±15.49ab

56.64±6.97ab

PP-1%

75.55±1.733a

56.70±0.46a

18.86±1.29ab

114.06±2.25a

57.36±1.80a

PP-3%

71.22±1.15a

51.44±1.18abc

19.78±0.25a

106.44±2.50abc

55.00±1.32ab

PP-5%

65.75±1.61ab

48.50±1.15bc

17.25±1.68bc

99.08± 2.04bc

50.58±2.30bc

PP-7%

61.42±1.09b

44.67±0.75c

16.75±0.66c

93.03± 1.40c

48.36±0.80c

Treat-ment

Hardness (g)

Springi-ness

Cohesi-veness

Gummi-ness

Chewi-ness

Control 1123.97±

48.56c 0.85± 0.06a

0.72± 0.02ab

811.32± 46.51c

688.15± 76.65b

PP-1% 1257.21± 165.55bc

0.87± 0.10a

0.73± 0.01a

853.88± 157.02bc

813.77± 182.77ab

PP-3% 1304.08±

51.11b 0.90± 0.03a

0.73± 0.01a

947.23± 35.64ab

945.39± 58.38a

PP-5% 1320.61± 120.51ab

0.90± 0.04a

0.72± 0.03ab

944.20± 93.59ab

848.62± 91.42a

PP-7% 1448.33±

77.74a 0.88± 0.01a

0.70± 0.02b

1004.40± 86.10a

850.76± 44.51a

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noodle was 69.11% in the control but gradually went down to 58.26% in the 7% treatment. And in the volume test, there were no differences between control and treatments except 7% treatment, which meant pork round powder had less capacity for water absorption [10]. Turbidity was tested in the cooked noodle soup to find the transferring effect of solid content from noodle to soup. With the higher contents of pork round powder in the noodle, the higher value of turbidity was resulted from 0.12 OD in the control and 1% treatment to 0.23 OD in the 7% treatment (p<0.05). All of these results causes for the weakness of binding capacity of noodle because the additives like meat powder replaces flour can't make up the combining strength enough when they are heated [11].

Table 5 Water absorption, volume and turbidity changes of cooked noodle with various ratio of

pork round powder addition

PP-1,3,5,7% : Pork Round Powder 1,3,5 and 7% addition a-d Mean with same letter in each column are not signifi-

cantly different by Ducan’s multiple range test (p<0.05).

Sensory Evaluation The result of sensory evaluation of cooked noodles added pork round powder is in Table 6. The sensory score in the color of 1% and 3% treatments showed higher than that of control, which was believed panels preferred the colored noodle than the white. Because they can think colored noodle contains functional material which helps health than just flour based white noodle. In the flavor and taste, there were similar results in the 1% and 3% treatment compare to control but different significantly in the 5% and 7% treatments (p<0.05). And there was no significant difference in the texture of the cooked noodle between 1%, 3% treatment and control. In the overall acceptability, the sensory score of 5% treatment was 5.3 but there was no significant difference compare to the score of 6.7 in the control. The sensory score in the 7% treatment was 4.8, which showed lowest result among

the treatments and also had significant different to the control (p<0.05). According to the result of this sensory evaluation, 3% of pork round powder treatment is found the best result among the treatments.

Table 6 Sensory evaluation of noodles with various ratio of pork round powder addition


Hedonic scale: 1 (extremely bad) to 9 (extremely good). a-c Mean with same letter in each column are not signifi-

cantly different by Ducan’s multiple range test (p<0.05).

IV. CONCLUSION Noodles added pork round powder 1%, 3 %, 5% and 7% respectively were manufactured and compared its quality characteristics with control. In the color test, control showed highest L and b value but a Value of 5% and 7% treatment had higher than other treatments significantly. In the test of viscosity, up to 3% treatment showed no difference compare to control and recognized it in the acceptable scope. In the hardness of texture profile test, it got higher score with more addition of pork round powder because it had different type of physical properties. Tensile force and tensile distance were important quality criteria in this experiment because of its binding strength of noodle. In this experiment, tensile force in the 3% treatment was 44.15g which showed similar result with the control of 44.65g (p<0.05). Turbidity went up while the pork round powder ratio was high which meant solid in the noodle extracted to the soup. In the color, flavor and taste test of sensory evaluation resulted reasonable in the 1% and 3% treatment. With these above results of test, 3% pork round powder treatment in the noodle is the best condition in this test because it remains reasonable ranges when it is compared to control. REFERENCES

1. Park, B. H., and Cho, H. S. (2006) Quality Characteristics Of Dried Noodles Made With Dioscorea Japonica Flour. Korean J. Food Cookery Sci. 22: 173-180

Treatment Water

absorption (%)

Volume (ml)

Turbidity of Soup (OD)

Control 69.11 70±0.02a 0.12±0.01d

PP-1% 68.26 70±0.01a 0.12±0.02d

PP-3% 61.71 70±0.02a 0.15±0.01c

PP-5% 60.49 70±0.02a 0.17±0.01b

PP-7% 58.26 70±0.05b 0.23±0.03a

Treat-ment

Color Flavor Taste Texture Overall

acceptability

Control 5.3±0.9b 6.2±1.3a 6.4±1.3a 6.9±1.0a 6.7±1.1a PP-1% 6.4±1.1a 6.2±1.2a 5.9±1.5ab 6.2±0.7ab 6.2±1.1a PP-3% 6.4±0.9a 5.7±1.2ab 5.7±1.1ab 6.3±1.0ab 5.7±1.3ab PP-5% 6.0±1.1ab 4.9±1.3b 4.9±1.6bc 5.3±0.5b 5.3±1.4ab PP-7% 6.0±0.9ab 4.4±1.3b 4.1±1.2c 5.1±0.7b 4.8±1.6b

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2. AACC. (2000) Approved Methods of the AACC. 10th ed. American Association of Cereal Chemists. St. Paul. MN. USA.

3. SAS Institute, Inc. (2002) SAS/STAT User’s Guide: Version 9.0. SAS Institute, Inc., Cary, North Caroline. USA.

4. Kim, S. Y., Kang, M. Y., and Kim, M. H. (2008) Quality Characteristics of Noodle Added With Browned Oak Mushroom (Lentinus edodes). Korean J.Food Cookery Sci. 24: 665-671

5. Kim, H. R., Hong, J. S., Chol, J. S., Han, G. J., Kim, T. Y., Kim, S. B., and Chun, H. K. (2005) Properties Of Wet Noodle Changed By The Addition Of Sanghwang Mushroom (Phellinus linteus) Powder And Extraet. Korean J. Food Sci. Technol. 37: 579-583

6. Sung, S. Y., Kim, M. H., and Kang, M. Y.

(2008) Quality Characteristics of Noodle Containing Pleurotus eryngii. Korean J. Food Cookery Sci. 24: 405-411

7. Lee, Y. S., Lim, N. Y., and Lee, K. H. (2000)

A Study on the Preparation and Evaluation of Dried Noodle Products made from Composite Flours utilizing Arrowroot Starch. Korean J. Soc. Food Sci. 16: 681-688

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Effects of Membrane-filtered Powder of Sunmul on the Quality Characteristics of Noodles. J Korean Soc. Food Sci. Nutr. 35: 199-204

9. Lee, Y. T., Jung, J. Y. (2003) Quality

Characteristics of Barley ß-Glucan enriched noodle. Korean J. Food Sci. Technol. 35: 405-409

10. Lee, K. H., and Kim, K. T. (2000) Properties of

Wet Noodle Changed by the Addition of Whey Powder. Korean J. Food Sci. Technol. 32: 1073-1078

11. Oh, B. Y., Lee, Y. S., Kim, Y. O., Kang, J. H.,

Jung, K. J., and Park, J. H. (2010) Quality Characteristics of Dried Noodles Prepared by Adding Hericium erinaceum Powder and Extract. Korean J. Food Sci. Technol. 42: 714-720

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RAPID AUTHENTICATION OF ENHANCED QUALITY PORK BY VISIBLE AND NEAR INFRARED SPECTROSCOPY

N. Prieto1,2*, M. Juárez2, R. T. Zijlstra1, Ó. López-Campos2,3, and J. L. Aalhus2

1 Department of Agricultural, Food and Nutritional Science, University of Alberta, 4-10 Agriculture/Forestry Centre, Edmonton,

Alberta, T6G 2P5, Canada. 2 Lacombe Research Centre, Agriculture and Agri-Food Canada, 6000 C&E Trail, Lacombe, Alberta, T4L 1W1, Canada.

3 Livestock Gentec, Edmonton, Alberta, T6G 2C8, Canada.

*[email protected]

Abstract – This study tested the ability of visible and near infrared spectroscopy (Vis-NIRS) to authenticate enhanced quality pork. One hundred and forty eight pigs from several genetic backgrounds, genders and diets were slaughtered at either 120 or 140 kg. Following splitting of the carcass, the right half carcass was blast chilled (BC) at -20 °C with a 2.5 m/sec wind speed for 1 h and moved into a cooler at 2 °C for 23 h, whereas the left half carcass was conventionally chilled at 2 °C for 24 h (Non-BC). The half loin from each left carcass side was moisture enhanced (ME; 0.50% salt and 0.49% disodium phosphate; pump rate 10%), whereas the other half was not subjected to ME treatment (Non-ME). Both ME and Non-ME half loins and another one from the right half carcass (BC) were cut in half, and the quarter loins were packed and aged for 2 or 14 days in a 1 °C cooler (2 and 14 d aged). The half and quarter loins were randomized by location to reduce location effects. After aging, Vis-NIR spectra were collected on the intact chops at the end of the loin using a portable LabSpec®4 spectrometer (350-2500 nm) at the laboratory. Partial least squares discriminant analysis based on Vis-NIR spectra correctly classified 90 and 95% of the 2 and 14 d aged pork samples. Vis-NIRS also correctly classified 99 and 96% of the Non-ME and ME pork samples aged for 2 d, and 95 and 94% of the Non-ME and ME samples aged for 14 d, respectively. Conversely, Vis-NIRS technology only correctly classified 57 and 54% of the Non-BC and BC samples aged for 2 d, and 53 and 54% of those aged for 14 d, respectively. Vis-NIRS technology can accurately discriminate the 14 d from 2 d aged and the ME from Non-ME pork samples but not the BC from Non-BC ones. I. INTRODUCTION To meet customer demands, pork processors are currently using processes such as moisture enhancement (ME) or blast chilling (BC) to

consistently produce a pork product with enhanced quality. Moisture enhanced meat is produced through multineedle injection of a brine solution that may contain ingredients such as salt, phosphates, sodium lactate, and lemon juice solids. The ME process produces a retail product with increased juiciness and tenderness and with substantially improved sensory quality [1]. BC can quickly reduce temperatures and has improved pork quality by lessening the incidence of PSE muscle [2]. Although some conflicting results exist, BC improved muscle color scores and firmness in some studies [3]. Additionally, it is well known that during meat aging, tenderness increases and characteristic flavors are developed [4]. In order to assure quality control and guarantee consumers that they are getting exactly what they paid for and not an inferior quality pork, rapid methods to distinguish pork products with enhanced quality are required. Near infrared spectroscopy (NIRS) is a sensitive, fast, and non-destructive technology, neither requiring reagents nor producing waste, that with minimal or no sample preparation provides information about the molecular bonds of organic compounds and tissue ultra-structure in a scanned sample [5]. NIRS has been successfully used for classification purposes in several species [6]. However, to the best of our knowledge, this technology was not to discriminate differentiated quality pork products. Therefore, the aim of the present study was to examine the potential of visible (Vis) and NIR spectroscopy to authenticate enhanced quality pork.

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II. MATERIALS AND METHODS A. Animals One hundred and forty eight pigs from several genetic backgrounds, genders, diets, and slaughter weights (3 × 2 × 3 × 2) were raised at the Lacombe Research Centre (LRC-Agriculture and Agri-Food Canada, Lacombe, AB, Canada). The genotypes were Duroc, Lacombe (Peak Swine Genetics, Leduc, AB, Canada) and Iberian (Semen Cardona, Cardona, Barcelona, Spain) sires × commercial Large White*Landrace F1 dams. Animals were fed a typically Canadian commercial diet, a high-oleic diet (canola based diet), or a high linolenic diet (flaxseed based diet) formulated to increase the omega-3 content in pork (O&T Farms, Ltd., Regina, SK, Canada). Pigs had ad libitum access to feed and water. All pigs in the study were managed, handled and slaughtered in accordance with the principles and guidelines established by the Canadian Council of Animal Care [7]. B. Sample collection and treatments When the animals reached the designated slaughter weight (either 120 or 140 kg), they were sent to the federally inspected abattoir at the LRC, stunned (400 V for 3 seconds), exsanguinated and dressed in a simulated commercial manner. Processing of carcasses included pasteurization (16 nozzles at 12 L/nozzle for 10 seconds with 86.4°C water for a total of 192 L/carcass) using an on-line stainless steel pasteurizing cabinet. Following splitting of the carcass, the right half carcass was blast chilled (BC) at -20 °C with a 2.5 m/sec wind speed for 1 h and moved into a cooler at 2 °C for 23 h. The left half carcass was conventionally chilled at 2 °C for 24 h (Non-BC). A half loin from each left carcass side was moisture enhanced (ME; 0.50% standard salt and 0.49% disodium phosphate; pump rate 10%; Hela Spice Canada Inc., Uxbridge, ON, Canada), whereas the other half was not subjected to ME treatment (Non-ME). Both ME and Non-ME half loins and another one from the right half carcass (BC) were cut in half, and the quarter loins were packed and aged for 2 or 14 days in a 1 °C cooler (2 and 14 d aged). The half and quarter loins were randomized by location to reduce location effects. After the corresponding ageing period, the chop at

the end of the loin from each quarter loin was used for collection of Vis-NIR spectra. C. Vis-NIR spectra collection A portable LabSpec®4 Standard-Res spectrometer (Analytical Spectral Device-ASD Inc., Boulder, CO, USA) equipped with an ASD fibre-optic high intensity contact probe (21 mm window diameter) was used to scan the intact chops at the laboratory. The spectrometer scanned 50 times per reading (~5 s) over the Vis-NIR range (350-2500 nm) in reflectance mode, and spectra were averaged by the equipment software. The data were interpolated to produce measurements in 1 nm steps, resulting in a diffuse reflectance spectrum of 2151 data points. Absorbance data were stored as log (1/R), where R was the energy reflected. Five spectra per chop were collected to increase the area of muscle scanned and reduce the sampling error (5), and then averaged. Instrument control and initial spectral manipulation were performed with the Indico™ Pro software package (Analytical Spectral Device-ASD Inc., Boulder, CO, USA). D. Statistical analysis Principal component analysis (PCA) was performed to decompose and compress the data matrix and to detect outlier samples. Partial least squares discriminant analysis (PLS2-DA, [8]) was applied on the raw spectra to classify samples into each of the treatments studied (2 and 14 d aged; Non-ME and ME; Non-BC and BC). This model seeks to correlate spectral variations (X) with defined classes (Y), attempting to maximize the covariance between the two types of variables for group differences and ignoring variance within a class. In this type of approach, Y is a dummy matrix with arbitrary numbers assigned to the different classes to be distinguished (2 d aged = 1, 14 d aged = 2; Non-ME = 1, ME = 2; Non-BC = 1, BC = 2). According to this equation, a sample was classified as meat belonging to a specific category (2 or 14 d aged; Non-ME or ME; Non-BC or BC) if the predicted value was within ±0.5 of the dummy value. The accuracy of the models obtained was evaluated using the percentage of correctly classified samples. Cross-validation was performed to validate calibrations and to restrict the number of PLS terms incorporated in the regression, to prevent over-fitting. Spectral data

300


management and PLS2-DA were performed by means of The Unscrambler® software (version 10.2, Camo, Trondheim, Norway). III. RESULTS AND DISCUSSION When the Vis-NIR spectra were collected on the non-BC and non-ME chops aged for 2 and 14 d, the regression model developed using a PLS2-DA and including 6 PLS terms correctly classified 90 and 95% of the pork samples, respectively (Table 1). Similar results were observed when the calibration model was cross-validated, where 10 and 7% of the pork samples aged for 2 and 14 d were misclassified, respectively. Regarding the spectra collection on the Non-ME and ME pork samples aged for 2 d, the discrimination model, including 7 PLS terms, correctly classified 99 and 96% of the samples, respectively. With regard to the validation, 1 and 10% of the pork samples from Non-ME and ME treatments were misclassified, respectively. When the Non-BC and BC samples were aged for 2 days, the PLS2-DA (1 PLS term) could only correctly discriminate 57 and 54% of the pork samples in the calibration, and 57 and 55% in the validation, respectively. This indicates that, by means of the Vis-NIR spectra, it was not possible to distinguish 2 d aged pork samples from BC carcasses from those conventionally chilled, since there would be about 50% chance of classifying a sample as BC or Non-BC.

Table 1 Discrimination results based on raw visible and near infrared spectra

Correctly classified (%)

Treatment Calibration Cross-Validation

2 d aged/14 d aged 90.0/95.0 90.0/92.8

Non-ME/ME (2 d aged) 99.3/96.3 99.3/89.7

Non-BC/BC (2 d aged) 57.4/54.4 56.8/55.0

Non-ME/ME (14 d aged) 95.2/94.4 93.8/91.6

Non-BC/BC (14 d aged) 52.7/54.1 50.0/52.7

ME: moisture enhancement; BC: blast chilling. When the quarter pork loins were aged for 14 d, the discrimination results for the Non-ME and ME pork samples were slightly lower than those reported for the 2 d aged ones; discrimination models (7 PLS) correctly classified over 95 and 94% of the Non-ME and 94 and 92% of the ME

samples in calibration and cross-validation processes, respectively. When the Non-BC and BC samples were aged for 14 d, about 50% of both pork samples were correctly classified (53 and 50% of the Non-BC and 54 and 53% of the BC pork samples during calibration and cross-validation, respectively; 1 PLS). Hence, Vis-NIRS technology was not successful to discriminate BC pork samples aged for 14 d from those conventionally chilled. Since the aging and ME processes entail changes in meat related to texture and water content, the successful Vis-NIRS performance in the discrimination of 2 from 14 d aged and Non-ME from ME pork samples could be due to the information related to water content and the structure of the muscle (i.e., the fibre arrangement of the muscle) obtained from the NIR region. Indeed, in Figure 1, differences between the 2 and 14 d aged and the Non-ME and ME (2 d aged) pork samples were observed in the NIR region due to the absorption of O-H bonds of water (970, 1450 and 1940 nm; [9]) and C-H bonds of fat (1215, 1725 and 1765 nm), the latter as a consequence of the inverse relationship between fat and water content in meat. Additionally, differences between the 2 and 14 d aged and between the Non-ME and ME (2 d aged) pork samples were found due to the N-H bonds of protein in the NIR region (2180, 2300, 2352 and 2470 nm) and the redox states of myoglobin in the Vis region (548, 580 and 762 nm; [10]), respectively. On the contrary, the spectra from the Non-BC and BC pork samples aged for 2 d were very similar, only showing minimal differences in the NIR region due to the O-H and C-H bonds absorption, which were not enough for Vis-NIRS technology to successfully distinguish between both samples. When the average spectra of the pork samples aged for 14 d were plotted (data not shown), differences between the Non-ME and ME samples were observed in the same regions than those found for the 2 d aged pork samples. However, no differences were found between the spectra from the Non-BC and BC samples.

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Fig. 1. Average Vis-NIR spectra (n = 148) of a) 2 and 14 d aged pork samples, b) moisture enhanced (ME)

and non-ME and c) blast chilled (BC) and non-BC pork samples aged for 2 d.

CONCLUSION Vis-NIRS technology has the potential to discriminate 2 from 14 d aged and moisture enhanced from non-moisture enhanced pork samples. Conversely, Vis-NIRS technology was not able to distinguish blast chilled pork samples from those conventionally chilled. This technology could hold value for on-line application in processing plants and at retail to authenticate pork of enhanced quality. ACKNOWLEDGEMENTS Financial support was provided by the Alberta Livestock and Meat Agency (ALMA). Drs. N. Prieto and Ó. López-Campos thank the Alberta Crop Industry Development Fund (ACIDF) and Livestock Gentec, the Canadian Beef Grading Agency and ALMA for their funding support, respectively. The authors also thank the Lacombe Research Centre operational, processing and technical staff for their dedication and expert assistance. REFERENCES 1. Brewer, M. S., Gusse, M. & McKeith, F. K. (1999).

Effects of injection of a dilute phosphate-salt solution on pork characteristics form PSE, normal and DFD carcasses. Journal of Food Quality 22: 3375-3385.

2. Milligan, S. D., Ramsey, C. B., Miller, M. F., Kaster, C. S. & Thompson, L. D. (1998). Resting of pigs and hot-fat trimming and accelerated chilling of carcasses to improve pork quality. Journal of Animal Science 76: 74-86.

3. Crenwelge, D. D., Terrell, R. N., Dutson, T. R.,

Smith, G. C. & Carpenter, Z. L. (1984). Effect of chilling method and electrical stimulation on pork quality. Journal of Animal Science 59: 697-705.

4. Lawrie, R. A. & Ledward, D. A. (2006). Lawrie’s

meat science. 6th ed. Cambridge: Woodhead Publishing, 336 pp.

5. Downey, G. & Hildrum, K. I. (2004). Analysis of

Meats. In L. Al-Amoodi, R. Craig, J. Workman, & J. Reeves III, Near-infrared spectroscopy in agriculture (pp 599-632). Winconsin: American Society of Agronomy Inc., Crop Science Society of America Inc., Soil Science Society of America Inc.

6. Prieto, N., Roehe, R., Lavín, P., Batten, G. &

Andrés, S. (2009). Application of near infrared reflectance spectroscopy to predict meat and meat products quality: A review. Meat Science 83: 175-186.

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Guide to the care and use of experimental animals. In: Canadian Council on Animal Care, Olfert, E.D., Cross, B.M., McWilliams, A.A., Ottawa Ontario, Canada. Volume 1.

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(2002). A user-friendly guide to multivariate calibration and classification. Chichester: NIR Publications, 420 pp.

9. Shenk, J. S., Westerhaus, M. O. & Workman, J. J.

(1992). Application of NIR spectroscopy to agricultural products. In D. A. Burns, & E. W. Ciurczak, Handbook of near infrared analysis. Practical spectroscopy series (pp 383-431). New York: Marcel Dekker.

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R. (2000). Study of dissected lamb muscles by visible and near infrared reflectance spectroscopy for composition assessment. Animal Science 70: 417-423.

a)

c)

b)

302


DIELECTRIC PROPERTIES OF SELECTED PORK ORGAN MEATS AND EXPERIMENTAL PATE PRODUCT FORMULATIONS FOR

CONTINUOUS FLOW MICROWAVE PROCESSING

Dana J. Hanson, Mehmet Kemal, Travis Tennant, Josip Simunovic

Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA

Abstract – Continuous flow microwave processing (CFMP) for pasteurization and sterilization is an emerging technology which has been recently commercialized for industrial production of vegetable, fruit and dairy products. There is a potential for use of CFMP for processing of specialty meat products such as pates and soft sausages with a high content of organ meats. The application of CFMP through production of specialty products for export may add value to these ingredients. I. INTRODUCTION Continuous flow microwave technology has been developed at North Carolina State University for sterilization of foods and biomaterials and has been commercially used over the last 5 years for production of vegetable and fruit based products (Coronel et al. 2005, Kumar et al. 2008, Steed et al. 2008). Processing of meat pates and other comminuted meat products that are commercial sterilization and thus stored and distributed under ambient (room) temperature conditions is common in Europe, Asia and other parts of the world. Currently this process represents a very small market segment in the United States. Processing and packaging of pate type products to achieve commercial sterility could be used to increase the consumption and particularly export potential of these products. Continuous flow microwave sterilization followed by aseptic packaging is one of the more recent technological developments which could provide the basis for development of these products.

In order to develop the continuous flow protocol for these new products, the dielectric properties need to be measured for all ingredients and potential formulations. These measurements have been used in both static (which are appropriate for homogeneous products) and continuous flow recirculated conditions (suited for heterogeneous and particulate-containing products), and have been reported by Coronel et al. (2008), Kumar et al. (2007) and Brinley et al. (2008). The dielectric property values have been subsequently used to develop the appropriate processes for preservation of a range of high acid and low acid products at pilot plant, semi-industrial and industrial capacity level. In order to initiate the development of appropriate processing sequences for continuous flow microwave sterilization a detailed dielectric property measurements and analyses are needed for each considered formulation and each of its major ingredients. This paper presents the methods used and results obtained for pork trim, organs as well as three different pate formulations within the realistic temperature ranges these products will need to be subjected to achieve commercial sterility. II. MATERIALS AND METHODS Raw meat samples (pork lean trimmings, pork liver and pork kidney) were ground through a 12mm grinder plate prior analysis. Treatment 1 represents ground pork lean trimmings, approximately 70% lean and 30% fat. Treatment 2 represents ground pork liver. Treatment 3 represents ground pork kidneys. Pork hearts were ground and used in the formulation of the

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pate, but they were not used as an individual treatment. This ground meat was then used in the following pate formulations. Treatment 4 meat block was a 1:1:2 ratio of pork liver, pork heart and 70% lean pork trimmings. Treatment 5 meat block was a 1:1 ratio of pork heart and 70% lean pork trimmings. Treatment 6 meat block was a 1:1 ratio of pork kidney and 70% lean pork trimmings. Pates were made using these meat blocks in combination of a standard spice block, including white pepper, coriander, nutmeg and onion powder. Salt was formulated at 2% of the meat block and all treatments were cured using 6.25% sodium nitrite curing salt at a 136ppm level. Raw mixed samples were emulsified by a Stephan Microcut MC machine (Stephan Machinery, Schwarzenbek, Germany). Raw samples were bagged and held at 4°C until further analysis. Cook samples were prepared by depositing raw pate in a plastic cooking bag and thermally process until an internal temperature of 71°C was achieved. Cooked pate samples were stored at 4°C until dielectric properties were recovered. Dielectric properties of the samples were measured from 10°C to 120°C using an open ended coaxial probe (HP 85070B, Agilent Technologies, Palo Alto, CA) connected to a network analyzer (HP 8753C, Agilent Technologies, Palo Alto, CA). The network analyzer was calibrated by leaving the tip of the probe in contact with air, metal, and 25ºC deionized water and measuring the dielectric properties. The dielectric properties were measured in the range of 300 to 3000MHz. The dielectric property measurement system is illustrated in Figure 1.

Figure 1. Instrument setup for measurement of dielectric properties in batch mode, with pressure cell and dielectric probe

Each sample was placed in a hermetically sealed stainless steel sample cup interfaced with the dielectric probe and incrementally heated in an oil bath, while taking the measurements at temperature increments of 10°C from 10°C to 120°C . From the obtained results, dielectric constant (ε'), dielectric factor ( ε”) and dielectric loss tangent have been plotted for each temperature level and for 2450 MHz and 915 MHz microwave frequencies, representing the two potential ranges to be used for eventual processing.

III. RESULTS AND DISCUSSION

Results for the dielectric properties of pork trim, liver and kidney are presented in Fig. 2 through Figure 4. Dielectric properties of three pate formulations at 915 MHz and 2450 MHz are presented in Fig. 5, 6 and 7.

Dielectric Properties of Lean Pork Trimmings at 2450MHz

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Figure 2. Dielectric properties of treatment 1, pork trimmings at 915 MHz and 2450 MHz at temperatures from 10C to 120C.

In all the analyzed cases, test ingredients and materials behaved similarly to other previously analyzed foods - dielectric constant decreased with the increasing temperature and dielectric factor increased with the increasing temperature. For analyzed test formulations, addition of salt and had a significant role, especially at 915MHz, in increasing the rate of the increase of dielectric loss tangent.

Dielectric Properties of Pork Liver at 2450MHz

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Figure 3. Dielectric properties of treatment 2, pork liver at 915 MHz and 2450 MHz at temperatures from 10C to 120C.

Dielectric Properties of Pork Kidney at 2450MHz

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Figure 4. Dielectric properties of treatment 3, pork kidney at 915 MHz and 2450 MHz at temperatures from 10C to120C.

Dielectric Properties of Treatment #2 at 2450MHz

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Figure 5. Dielectric properties of treatment 4, at 915 MHz and 2450 MHz at temperatures from 10C to 120C.


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Figure 7. Dielectric properties of treatment 6 at 915 MHz and 2450 MHz at temperatures from 10C to 120C.

The increase in the dielectric loss tangent is usually associated with an increase in the rate of heating and reduction of penetration of microwaves into the processed material. This decreased depth of penetration results in the loss of heating in deeper segments of the material and occasional overheating around the edge of the flow-through conduits under continuous flow conditions. While at 2450 MHz this trend was not as prominent as at 915, it should be taken into consideration when designing the processing systems for these types of products in the future. Reduction of salt content in experimental formulations could be potentially prove to be beneficial for the uniformity of product heating under continuous flow conditions, but would need to be confirmed both via additional dielectric property measurements for optimized formulations as well as experimentally under continuous flow heating and sterilization. These initial results indicate that the 2450 MHz microwave frequency might also be appropriate for processing of these types of products, but this also needs to be tested experimentally in future studies.

IV. CONCLUSION

Dielectric properties of pork trim, organs and pork pate products have been measured and evaluated for potential compatibility with the emerging continuous flow microwave technology for thermal sterilization of foods. Properties were measured for both industrially relevant microwave frequencies (915 MHz and 2450 MHz) and within a temperature range encompassing refrigeration (10C) to sterilization (120C). Test material properties followed the general trends observed previously for other foods. The addition of salt was related to the increase in dielectric loss tangent in test formulations, especially in the 915MHz range. Further experimental studies are recommended to define the processing parameters and ranges for these products more precisely.

V. REFERENCES 1. Coronel, P., Truong, V.D., Simunovic, J., Sandeep, K.P. (2005). Aseptic processing of sweetpotato purees using a continuous flow microwave system. Journal of Food Science. Vol. 70(9): E531-E536. 2. Kumar, P., Coronel, P., Truong, V.D., Simunovic, J., Swartzel, K.R., Sandeep, K.P. , Cartwright, G.D. (2008). Overcoming issues associated with the scale-up of a continuous flow microwave system for aseptic processing of vegetables purees. Food Research International. Vol. 41(5): 454-461. 3. Steed, L., Truong, V.D., Simunovic, J., Sandeep, K.P., Kumar, P., Cartwright, G.D., Swartzel, K.R. (2008). Continuous flow microwave-assisted processing and aseptic packaging of purple-fleshed sweetpotato purees. Journal of Food Science. Vol. 73(9): E455-E462.

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4. Coronel, P., Simunovic, J., Sandeep, K.P., Kumar, P. 2008. Dielectric properties of pumpable food materials at 915 MHz. International Journal of Food Properties. Vol. 11(3): 508-518. 5. Kumar, P., Coronel, P. Simunovic, J., Sandeep, K.P. 2007. Measurement of dielectric properties of pumpable food materials under static and continuous flow conditions. Journal of Food Science. Vol. 72(4): E177-E183 6. Brinley, T., Truong, V.D., Coronel, P., Simunovic, J., Sandeep, K.P. 2008. Dielectric properties of sweetpotato puree at 915 MHz as affected by temperature and chemical composition. International Journal of Food Properties. Vol. 11(1): 158-172.

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THE EFFECT OF BRINE INJECTION LEVEL ON MOISTURE RETENTION AND SENSORY PROPERTIES OF

CHICKEN BREAST MEAT

E. Moholisa1, E. Roodt1, C. Bothma1, F. de Witt2 , and A. Hugo1*,

1Department of Microbial, Biochemical and Food Biotechnology, University of Free-State, Bloemfontein, South Africa 2Department of Animal, Wildlife and Grassland Science, University of Free-State, Bloemfontein. South Africa

*[email protected]

Abstract – This study was conducted to determine the effect of 15% and 30% brine injection levels on chicken breast quality. Breasts injected to 15% showed lower thawing and cooking losses, compared to control and 30 % injected breasts. Control breasts had significantly (p<0.001) higher cooking losses compared to brine injected breasts, which could be ascribed to the absence of the brine ingredients, which held the moisture during cooking. Sensory properties of chicken meat, injected to 15% and 30% brine, were compared to uninjected controls by a 75 member consumer panel. The brine injected breasts were significantly preferred to the control samples, while the 15% injection level breast cuts were significantly preferred to the 30% level breast cuts. Tenderness of cooked meat was instrumentally predicted by Warner-Bratzler shear (WBS). The shear force values were significantly lower in brine injected breasts than controls. Sensory panellists rated breasts, injected to 15%, significantly more tender than 30% brine injected breasts. I. INTRODUCTION During commercial processing of poultry, brine is often introduced into poultry meat through injection. This has been justified on various technological grounds, such as improving the moisture retention and sensory properties (tenderness, juiciness, and flavour) [1, 2]. When brine injection was introduced, it was injected into poultry meat with an aqueous monosodium glutamate solution, to an extension level of 3% 8% of the weight of the dressed chicken [3]. But lately, chicken processors tend to inject brine at varying levels. Sodium chloride and phosphate are being used as basic ingredients, together with various other ingredients, such as antioxidants,

flavours, starch, non-meat proteins and hydrocolloids. In South Africa, injection levels are ranging from 30% 60% [4]. This is the result of no current legal regulations, governing brine injection levels into poultry meat, in South Africa [5].This lack of regulations results in poultry processors injecting brine into meat at varying percentages. It is important to emphasize that injection percentage is one of the most important factors that influences the quality of brine injected poultry meat. High injection levels may degrade some of the quality characteristics, such as texture and flavour, and may lead to excessive muscle contraction during cooking [2]. Information is still lacking on realistic injection levels to obtain optimum positive effects on meat quality.

Therefore, the aim of this study was to determine the effect of 15% and 30% injection levels on moisture retention and sensory properties of chicken breast. II. MATERIALS AND METHODS Sample preparation A total number of 288 “skin on” chicken breasts of known origin, not subjected to any processing steps that could add water to the portions, were injected with brine. Brine was prepared from cold tap water containing ice. Brine composition was calculated as follows,

Brine composition was altered to allow for a constant ingredient level in the product. Brine ingredient in product was 1% salt, 0.5% dextrose, 0.2% carrageenan, 0.1 % xanthan and 0.5% sodium tripolyphosphate. The chicken

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breasts with skin were injected manually. The treatments were control (no injection), 15% injection over raw weight, and 30% injection over raw weight. Breasts were weighed before, during and after injection, to determine the brine pick-up. After injection, breasts were vacuum packaged and stored at -18oC. Twelve portions were used for each treatment group, for all analysis.

Analytical procedures Total moisture loss: All the chicken portions were weighed pre- and post-thawing, as well as after cooking, for the determination of differences in the losses (%) during the thawing and cooking processes, respectively. Total moisture loss was calculated as:

Thawing losses: Chicken breast pieces were thawed at 4oC for 24 hours. Thawing loss was calculated as:

Cooking losses: Samples were subsequently dry cooked at 160°C to an internal end point temperature of 85°C, for approximately one hour in the oven (Mielé, model H217). Internal temperature was recorded according to the American Meat Science Association (AMSA) [6]. Calculations were performed according to the guidelines of the AMSA [7] and using Microsoft Excel 2007. Cooking loss was calculated as:

Shear force measurement: Cooked breast samples were cooled at 4°C overnight. Samples were cored, after being cooled further to room temperature (centrally controlled at 22°C). Cylindrical cores, with a diameter of 12.7 mm (20 – 22 mm long), were obtained from the mid-portions of the cooked muscle. Samples were sheared perpendicular to the fibre direction, with a Warner-Bratzler shear (WBS) device mounted on a Universal Instron Machine (Model 4301; Instron Corporation, 1990). The shear force was determined using 200 mm/min test speed with

a 1 kN load cell [8, 9, 10]. A total of 36 cores per treatment were obtained. Sensory analysis: Sample preparation was done according to the research guidelines of the AMSA [6]. During cooking, salt was lightly added to the control portions to compensate for the brine injected controls. The meat was evaluated in two sessions for the preference and attribute rating scale tests, respectively. Both tests were judged on a nine point hedonic scale, ranging from 1= dislike extremely to 9 = like extremely. The attributes tested were taste, tenderness, juiciness and aftertaste. For tenderness, the scale was changed to ‘extremely tough=1 to extremely tender=9’; for juiciness, to ‘extremely dry=1 to extremely juicy=9’ and for aftertaste from ‘non-present=1’ to ‘present=9’. A 75-member consumer panel, consisting of regular eaters of chicken meat, was used to evaluate the samples. Statistical analysis: Differences between treatments were determined, using a one-way analysis of variance (ANOVA) procedure. When applicable, the Tukey-Kramer multiple comparison test (α =0.05) was used to determine differences between treatment means [11]. III. RESULTS AND DISCUSSION The control treatment had significantly (p<0.001) higher thawing losses than the breasts injected to 15%. However, the thawing losses for the control treatment were significantly (p<0.001) lower than the breasts injected to 30% (Table 1). It was apparent that at higher injection levels the muscle fibres could not hold the excess water injected into meat. After cooking, the control treatment lost 28% water, compared to 24% and 25% of breasts injected to 15% and 30%, respectively. This confirmed that brine ingredients were able to keep water in the meat during cooking. Salt, carrageenan, xanthan and phosphate have previously been reported to retain water in meat [12]. Hydrocolloids possess a great gelling capacity, thereby contributing to an increased WHC (water holding capacity) during cooking [2]. Total moisture loss of the controls was significantly higher than that of

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Table 1: The effect of injection level on thawing loss, cooking loss and shear force resistance on

chicken breast meat.

a,b, Means with different superscripts in the same row differ significantly. the two injection levels. Even though not statistically significant when comparing the two injection levels, the cooking losses for the breasts injected to 15% were lower than the breasts injected to 30%. Volpato et al. [13] used 12% and 15% brine in the processing of deboned chicken breasts. They found that the use of 12% brine, in relation to the weight of the raw material, resulted in a lower water loss during cooking, than the use of 15% brine. More water loss by non-injected controls could be expected.

The WBS values were significantly (p<0.001) lower in brine injected breasts, than the control breasts that were not injected (Table 1). The shear values were the lowest with a 30% injection level, compared to the 15% injection level. This could be attributed to the gelling properties of carrageenan and xanthan gum. Dransfield [14] showed that injected substances were probably only effective up to limited injection (± 10%) and further improvement of tenderness, with higher levels of injection, only occurred due to the dilution effect of the water-hydrocolloid gel, which caused other obvious negative quality defects. Lower shear force values found in brine injected controls were in agreement with Baumert & Mandigo, Xargayó [1, 2]. They found that brine injected meat resulted in lower shear values when compared to controls. It is also important to realize that shearforce values as low as the ± 2.4 kg as observed in the control breasts can also be considered as very tender.

The control samples had significantly (p<0.001) lower sensory scores, than injected samples for both preference and attribute tests (Table 2). Breasts injected to 15% were preferred to 30% brine injected breasts. This could be expected, because brine ingredients had been shown to increase the sensory properties of meat [2]. The non-injected control breasts had lost water during thawing and cooking, thus contributing to less tender and less juicy meat. Although not statistically significant, breasts injected to 15% were preferred and rated even higher than breasts injected to 30%, for all the attributes,. These current findings are also consistent with that of Xargayó et al. [2], who found that products injected to 15% scored higher in overall acceptability, than at higher injection levels of 25%. The explanation, for the aftertaste being rated higher in brine injected samples, might be because of the added brine ingredients, meaning that the consumers could detect the added brine ingredients. Recommended injection percentages range between 5% and 20%, for increasing the meat’s sensory quality, depending on the type of animal and muscle [2]. Table 2: The effect of injection level on

sensory score of chicken breast meat.

Control 15%

Injection 30%

Injection Significance level

Preference 5.23a 6.80 c 6.13b p<0.001

Taste 4.38a 6.70b 6.63b p<0.001

Tenderness 4.04a 7.33b 7.03b p<0.001

Juiciness 3.29a 6.63b 6.53b p<0.001

Aftertaste 4.62a 6.32b 6.30 b p<0.001 a,b, Means with different superscripts in the same row differ significantly. IV. CONCLUSIONS

The effect of carrageenan and xanthan was apparent in keeping the moisture in the meat, during thawing and cooking of brine injected chicken portions. Brine injected meat was mostly preferred by the consumers. Sensory properties, which included taste, tenderness and juiciness, were enhanced in brine injected meat. The 15% injection level was preferred by consumers to the 30% injection level. These results showed that low brine injection levels induce positive effects on meat quality,

Control 15%

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Injection Significance

level Thawing loss (%)

4.04b 2.36 a 5.29c p<0.001

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28.41b 24.05 a 25.30 a p<0.001

Total moisture loss (%)

31.61c 25.85a 29.30b p<0.001

Shear force (kg)

2.39c 1.25b 1.06a p<0.001

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which were acceptable to the consumers. Results from this research clearly indicate that there is no real advantage in using injection levels higher than 15 % in fresh chicken. ACKNOWLEDGEMENTS E Moholisa acknowledges National Research Foundation (NRF) bursary and TATA Africa scholarships for financial support during her studies. REFERENCES 1. Baumert, J. L. & Mandigo, R. W. (2005). The

effects of phosphate type and potassium lactate on quality characteristics of enhanced beef steaks. Nebraska Beef Report (pp 95-99), University of Nebraska.

2. Xargayó, M., Lagares, J., Fernández, E., Borrell, D., & Juncà, G. (2010). Spray marinating: A definitive solution for improving meat texture. www.metalquimia.com/pdfs/doctec26.pdf. Retrieved on 05/02/2011.

3. Buchanan, B. F. (1955). US Patent 2, 709, 658. Process for treating poultry, Serial No. 221, 537. 17 April 1955. New York: International Minerals & Chemical Corporation.

4. Department of Agriculture, Forestry and Fisheries. (2011). Brine injection project and Supreme Poultry visit. www.daff.gov.za/doaDev/articles/BrineInjectionProject.html. Retrieved on 03/03/2011.

5. National Consumer Forum. (2010). Bloated chicken prices. Consumer Fair, Edition 4, 2010.

6. AMSA. (1995). Research Guidelines for Cookery, Sensory Evaluation of Meat and Instrumental Tenderness Measurements of Fresh Meat. Chicago. Illinois. National Livestock and Meat Board.

7. AMSA. (1978). Guidelines for Cookery and Sensory Evaluation of Meat. Chicago. Illinois. National Livestock and Meat Board.

8. Jaturasitha, S., Leangwunta, V., Leotaragul, A. & Phonghaew, T. (2002). A comparative study of Thai native chicken and broiler on productive performance, carcass and meat quality. Paper at Deutscher Tropentag, Witzenhausen. The Conference on International Agricultural Research for Development. October 9-11, 2002.

9. Castellini, C., Dal Bosco, A., Mugnai, C. & Pedrazzoli, M. (2005). Comparison of two chicken genotypes organically reared: oxidative stability and other qualitative traits of the meat. Italian Journal of Animal Science 5: 335-363.

10. Musa, H. H., Chen, G. H., Cheng, E. S., Shuiep, E.S. & Bao, W.B. (2006). Breed and sex on

meat quality of chicken. International Journal of Poultry Science 5: 566-568.

11. NCSS. (2007). Statistical System for Windows. Number Cruncher Statistical Systems. Kaysville, Utah, USA.

12. Feiner, G. (2006). Meat Products Handbook: Practical Science and Technology. London: Woodhead Publishing.

13. Volpato, G., Michielin, E. M. Z., Ferreira, S.R.S. & Petrus, J. C. C. (2007). Optimisation of the chicken breast cooking process. Journal of Food Engineering 84: 576-581.

14. Dransfield, E. (1994). Optimisation of tenderisation, ageing and tenderness. Meat Science 36: 105-121.

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GENETIC POLYMORPHISM OF ryr 1 GENE FROM TURKEY PSE BREAST MUSCLE.

Fernanda Gonzales Paião1*, Viviane Ribeirete2, Rafael Humberto de Carvalho3, Mayka

Reghiany Pedrão1, Fábio Augusto Garcia Coró1 and Massami Shimokomaki1,3

1 Federal Technological University - Paraná - Campus Londrina, Estrada dos Pioneiros, 3131,CEP 86036-370, Londrina. Paraná. Brazil.

2 Graduate Student, Federal Technological University – Paraná – Campus Londrina. Paraná. Brazil 3 Londrina State University –Londrina, PR 445, s/n, CEP 86051-990, Londrina. Paraná. Brazil

*[email protected]

Abstract – A mutation in ryr 1 gene affect meat quality in pork, causing PSE (Pale, Soft and Exudative) meat. The objective of this study was to identify polymorphisms in turkey´s α-ryr gene that could be associated with PSE meat. Because ryr genes are over 100,000 bp long and code for proteins with about 5000 amino acids, the primers used in this work amplified a portion of this gene, corresponding to the hotspot region 1 that contains the known mutation leading to PSE meat in pork. Total blood DNA was extracted from 10 breast muscles, 5 samples from PSE meat and 5 from samples considered normal. These DNA samples were amplified by PCR, cloned, sequenced, and used to identify single nucleotide polymorphisms (SNPs). The amplified fragment of α-ryr was 653 nucleotides in length. A non-synonymous nucleotide substitution (A/G) was identified in 3 PSE meats and 1 normal meat. This SNP caused a change from Met to Val in the α-RYR protein. Since this SNP was identified in both meat samples and due to the fact that this mutation does not change the structure and/or function of the muscle protein, this alteration in DNA sequence is an inappropriate candidate to be used as a genetic marker for turkey PSE meat. I. INTRODUCTION Porcine stress syndrome (PSS) leads to meat that is paler, tougher, and have lower water-holding capacity known as PSE meat [1]. The PSE meat results in great losses in the meat industry by offering product with altered colors and flavors and lower yield during cooking [2]. The main cause of PSS is a mutation at nucleotide 1843 of cDNA (GenBank M911451) of the gene encoding the calcium release channel in the skeletal muscle sarcoplasmic reticulum called ryanodine receptor (RYR1) commonly known as the halothane gene (HAL) [3]. Some turkey and chicken breast muscle have been found to be lighter or paler than what is considered to be normal [4, 5]. Similar to PSE pork, turkey pale

poultry muscles have reduced water-holding capacity (WHC) and higher drip loss than normal muscles. However, the lighter color poultry may also have normal WHC and drip loss [4,6]. Based on these findings, researchers have adopted the PSE term to describe pale avian muscle. However, the genetic basis for the PSE syndrome remains unknown and the participation of RYR on this abnormality has not yet been fully related to the occurrence of PSE meat in poultry [7,8,9]. The objective of this study was to identify genetic polymorphisms in a fragment of α-ryr homologous to mammalian ryr 1, sequencing a fragment of this amplified gene from normal and PSE turkey breast meats collected in a commercial plant in Brazil. II. MATERIALS AND METHODS

Turkeys 137- to 145-days-old were slaughtered under commercial processing plant conditions. The color (L values) and pH were measured in 810 breast fillet after 24h postmortem. Muscle samples (0.5 x 2.0 x 1.0 cm3) from 5 PSE fillets (pH< 5.7 and L*≥55.0) and 5 normal fillets (pH≥ 5.7 and L*<55) were collected and stored in microtubes (1.5 mL) at -20°C. Total DNA was macerated under nitrogen and extracted from Pectoralis major m. using the Axyprep multisource Genomic DNA Miniprep Kit (Axygen, Union City, CA) following the manufacturer's instructions. For PCR amplification, the primers were the same used by Droval et al. [8]. The PCR amplification consisted of an initial denaturation at 95ºC for 1 min, followed by 35 cycles at 95ºC for 30s, at 52ºC for 30s, at 68ºC for 2 min, and a final extension at 72ºC for 7 min. PCR confirmation was conducted by agarose gel electrophoresis and the amplified fragments were cut from the gel and purified

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using the Pure Link Quick Gel Extraction Kit (Invitrogen, Carlsbad, CA) following the manufacturer's instructions. All PCR products were inserted into the pGEM-T Easy ® vector (Promega, Madison WI) following the manufacturer's instructions and transformed using DH5a eletrocompetent cells. The recombinant clones were isolated and sequenced on both strands using the M13 universal primers and the BigDye® Terminator v3.1. kit (Applied Biosystems, Warrington, UK) for the automatic sequencer ABI 3100 (Applied Biosystems, Warrington, UK). The chromatograms obtained were manually analyzed using the program Vector NTI Suite 8 (InforMax), followed by removal of the vector sequence (Vector Screen (http://www.ncbi.nlm.nih.gov/VecScreen/VecScreen.html), and finally global alignment using ClustalW2 (http://www.ebi.ac.uk/Tools/clustalw2/index.html) to obtain the consensus sequence.

III. RESULTS AND DISCUSSION To identify the possible mutations, a region of 653 pb from ryr gene was amplified (Fig 1). This amplified fragment corresponded to the 3’ end of mutation hot spot 1 of human RYR1.

Figure 1:Electrophoresis of amplified products on a 1% agarose gel. Lane M=100-bp molecular weight marker (Invitrogen); lanes 1 and-2: DNA samples from PSE turkey meat; lanes 3 and 4: DNA samples from turkey normal meat, lane 6: negative control (no DNA). The ryr amplified fragment was cut from agarose gel, cloned and sequenced, showing 88% sequence identity with turkey RYR cDNA. Aligning the turkey α-ryr genomic sequence of allele I with the turkey cDNA sequence (EU177005.1) using the bl2seq

program [10], it was identified a complete intron localized in nucleotides 46-466. The similarity in 88% occurred because our sequence contains an intron, which is removed from RNAm during transcriptional process. This intron was not similar to ryr cDNA sequence available in Genbank database, because this cDNA was processed and the presence of intron was not observed. The part of our sequence was comparatively 100% in relation to the exons from turkey available in Genbank [11]. It was also identified two sequences similar to α-ryr that were called alleles I and II, respectively. A Single Nucleotide Polymorphism (SNP) was identified at position 612, where it was verified to be a substitution of an A (adenine) for a G (guanidine). Genomic sequence from allele I was more common within the samples with 60% of the birds being homozygous for A at position 612 (Fig. 2). Allele II, which carried the G to A substitution nucleotide at position 612, altered the RYR amino acid sequence relative to the turkey α-ryr, changing a methionine to a valine residue. According to the SIFT program [12] this substitution may cause changes within the protein structure, however this change may be tolerable, because both are hydrophobic amino acids. However, this detected polymorphism cannot be associated as mutation consequently promoting the occurrence of PSE meat, because in muscle that developed normal meat this phenomena was also detected. These results were similar to chickens [8] and corroborated to those reported by Chiang et al [7] suggesting that in poultry the ryanodine genetic basis of PSE meat is rather different from pork as recently suggested by Malila et al., [13].

Figure 2. Alignment of the partial nucleotide sequences from five turkey that carried a

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http://www.ncbi.nlm.nih.gov/VecScreen/VecScreen.html

http://www.ebi.ac.uk/Tools/clustalw2/index.html

http://www.ebi.ac.uk/Tools/clustalw2/index.html


polymorphism, both genomic sequences (alleles I and II) at position 612 of the α-RyR gene fragment. The polymorphic nucleotide position within each sequence is shown in bold.

IV. CONCLUSION

A polymorphic SNP was detected in α-ryr DNA sequences from turkey that developed either normal or PSE meat. However, this alteration in ryr gene was not conclusive to explain the turkey PSE meat genetic basis. ACKNOWLEDGEMENTS The authors are thankful to CNPQ, CAPES and UTFPR for supporting this work. FGP is pos doctoral CAPES research fellow; FAGC, MRP and MS are CNPq Research fellows. VR holds CNPq/IC scholarship. REFERENCES 1. Candek-Potokar, M. et al. (1998). Effects of age

and/or weight at slaughter on longissimus dorsi muscle: biochemical traits and sensory quality in pigs. Meat Science 48: 287-300.

2. O'Neill, D.J. et al. (2003). Effects of PSE on the quality of cooked hams. Meat Science 64: 113-118. Meat Science 64: 113–118.

3. Fujii, J. et al (1998). Identification of a mutation in porcine ryanodine receptor associated with malignant hyperthermia. Science 253: 448-451.

4. Owens, C.M. et al. (2000). The characterization and incidence of pale, soft and exudative turkey meat in commercial plant. Poultry Science 79: 553-558.

5. Marchi, D.F. et al. (2009) Sensitivity to halothane and its relationship to the development of PSE (Pale, Soft, Exudative) meat in female lineage broilers. Brazilian Archives of Biotechnology 52: 229-223.

6. Carvalho, R.H. et al. (2013) Commercial turkey PSE (Pale, Soft and Exudative) meat in Brazil during summer season. In Proceedings 59th International Congress of Meat Science and Technology (pp. 138-139), 18-23 August 2013, Izmir, Turkey.

7. Chiang, W. et al., (2003). Identification of two αRYR alleles and characterization of αRYR transcript variants in turkey skeletal muscle. Gene 330: 177-184.

8. Droval, A.A. (2012) A new single nucleotide

polymorphism in the ryanodine gene of chicken skeletal muscle. and Molecular Research 11: 821-829.

9. Paião, F.G, et al. (2013). Skeletal muscle calcium

channel ryanodine and the development of pale, soft and exudative meat in poultry. Genetics and Molecular Research 12: 3017-3027.

10. Altschul, S.F., et al. (1997). Basic local

alignment search tool. Jourrnal of Molecular Biology 215: 403-410.

11. Genebank site, disponible at:

http://www.ncbi.nlm.nih.gov/genbank/.

12. SIFIT program, disponible at: http://sift.bii.a-star.edu.sg/.

13. Malila, Y. et al. (2013). Differential gene

expression between normal and pale, soft and exudative turkey meat. Poultry Science 92:1621-1633.

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RESTRUCTURED AND BREADED (NUGGETS) CHICKEN PRODUCT WITH THE ADDITION OF OAT FIBER: CHEMICAL AND

SENSORIAL CHARACTERIZATION

Cristiane Fiorentin1, Mayka Reghiany Pedrão1, André Gallo2, Cézar Augusto Iziquiel2,

Magali Aparecida Mafra2 and Fábio Augusto Garcia Coró1

1 Professional Master Program, Federal Technological University - Paraná - Campus Londrina, Estrada dos Pioneiros, 3131,CEP 86036-370, Londrina. Paraná. Brazil.

2 Graduate Student, Federal Technological University – Paraná – Campus Londrina. Paraná. Brazil

*[email protected]

Abstract – The aim of this work was to add Oat Fiber (OF) in a restructured and breaded chicken product and determine their chemical composition, dietary fiber and sensory analysis of formulations. It was observed for chemical composition of products in different concentrations of OF (3%, 4.5% and 6%) that an increase in addition of fiber resulted in a decrease in the protein and moisture, whereas the opposite was observed for lipids and ashes, presenting significant difference (p <0.05) in the analysis of moisture, ash and protein. The content of dietary fiber in product followed the percentage of OF added in the formulations of 3% and 4.5%, but 6% did not correspond to the added percentage. In the analysis of color significant difference was observed (p <0.05) among the samples for parameters L* and b*, however there was no significant difference (p>0.05) for the parameter a*. Sensory analysis showed that the three formulations are similar to each other, with no significant difference (p> 0.05) for the attributes studied, only for juiciness significant difference was observed (p <0.05). This work has proved to be possible the application of dietary fibers in restructured and breaded meat products with a good overall acceptability by consumers. I. INTRODUCTION The industry have focused in producing food with high nutritional value by adding alternative ingredients. These ingredients need to satisfy this demand without interfering with two other important issues: product quality and consumer health, which is becoming more aware and critical of their food. Breaded and restructured meat products represent a category with great potential for application of functional ingredients with appeals of healthiness. This kind of products is very well accepted by the population, because its great convenience. Aditional studies are always needed in searching of improvements of technological, sensory and functional effects

of the ingredients used in such products. This segment of functional products, with the addition of fibers, is considered the new frontier in the food industry and one of the alternatives to add value to ingredients considered by-products in the industry. The presence of fibers in food often results in the reduction of calories and contributes to the amount of daily intake, thereby providing a better diet. According to the resolution of ANVISA (Brazilian National Health Surveillance Agency), which provides for the technical regulation on supplementary nutrition, we can consider a product as "source of fiber" when presenting in its composition at least 3g of fiber per 100g of prepared dishes or at least 2.5g of fiber per serving. For designate a product as "high fiber content" must have at least 6g of fiber in cooked dishes or at least 5g of fiber per serving BRAZIL [1]. The addition of an ingredient processing and breaded products may directly influence the quality attributes such as color, aroma, flavor, juiciness and tenderness. Quality aspects of a new product will determine their success and pleasant aroma and flavor, desirable color and texture become prerequisites. Therefore, knowledge of their composition, structure, process conditions and how these factors will influence the sensory aspects, nutritional value, safety, the costs and marketing share when developing a new product are extremely required. II. MATERIALS AND METHODS The process of elaboration of a restructured chicken meat product (nuggets) with the addition of OF was carried out in the laboratories of meat science of Federal Technological University – Paraná (UTFPR). The OF obtained by microgrinding was

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provided by SL Alimentos Company. The covering system (Predust, Breading and Batter) was supplied by Grififth Laboratories - GL. Other condiments incorporated into the formulation were purchased in local shops. The nuggets were processed by mixing the ingredients with the OF followed by molding and subsequent application of the cover layer (predust, batter and breading), pre frying at 200ºC for 25 seconds and freezing at -18°C. Three different formulations were prepared for analytical assays and shown below. Formulation 1: Standard (no OF added); Formulation 2: 3% of Oat Fiber added Formulation 3: 4.5% of Oat Fiber added Formulation 4: 6% of Oat Fiber added To determine the chemical composition of the formulations, methodologies recommended by the AOAC [2] were used. For dietary fiber, moisture, ash, lipids and proteins, were used the methods described in topics 941.43, 950.46, 920 153, 991.36 and 992.15, respectively. To the color analysis, were used the Minolta CR400 colorimeter, calibrated and expressed in the CIELAB system according Soares [3]. All the analysis were carried out in triplicate and for each treatment, e.g. concentration of OF, 20 samples were taken. To perform the sensory analysis was used Multiple Comparison Method [4] with 65 untrained panelists using a structured scale to scores from 1 to 5. All data were submitted to analysis of variance (ANOVA) and Tukey test at 5% using the Portable Software Statistica 8. III. RESULTS AND DISCUSSION The chemical composition of chicken nuggets prepared with different concentrations of OF are shown in Table 1. It was observed that the addition of OF altered the chemical composition of the product, causing a decrease in protein and moisture content. However, for lipid and ash content the opposite happened. The amount of protein decreased the extent that the fiber is added, resulting in a significant difference (p <0.05) between the standard and the samples added with 4.5% and 6% OF. Fact caused due to the substitution of chicken for OF. The larger the added fiber, lowest was protein concentration found in samples of breaded chicken, being in

accordance with the Technical Rules of Identity and Quality of Breaded [5], which states that the breaded meat products must have at least 10% protein and a maximum of 30% carbohydrates. The table below shows the results of total dietary fiber in samples of breaded chicken with the addition of OF. Table 1 Chemical composition of chicken nuggets added with different concentrations of Oat Fiber Std 3% 4.5% 6% Moisture 56.68

± 0.51 a 52.59

± 0.99 b 54.06

± 1.26 b 52.13

± 1.00 b Ashes 2.20

± 0.08 b 2.42

± 0.37 a,b 2.31

± 0.04 a,b 2.67

± 0.20 a Lipids 6.71

± 0.46 a 7.37

± 0.19 a 7.19

± 0.04 a 7.45

± 0.73 a Protein 17.25

± 1.64 a 15.99

± 0,74 a,b 14.04

± 0.74 b 13.18

± 1.48 b Carbs* 16.99 21.63 22.40 24.57 Values in the same line with different letters indicate significant difference (p <0.05). * Carbohydrates were calculated by difference. Std - Standard Table 2 Content of dietary fiber found in the formulations of breaded chicken with addition of OF.

Samples Dietary Fiber

(g/100g)

Std* <0,50 3% 3,09

4,5% 4,22 6% 4,54

*Std – Standard It is observed that dietary fiber content analyzed followed OF percentage in formulations added with 3% and 4.5%. However, the formulation with 6% OF did not match the added percentage, but remained with the highest dietary fiber level. The samples attended the Technical Regulation on Nutritional Supplement, containing at least 3% of dietary fiber per 100g, the labeling could receive the "source of fiber" attribute. From Table 3, we observe a significant difference between the samples analyzed for parameters L * and b *. However there was no significant difference for the parameter a*, which indicates that the higher the value, the color of the sample will tend to red. The higher the luminosity value (L*), the clearer the sample. The parameter b* denotes the amount of yellow color which in meats and their derivatives is related to brown.

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For the parameter L*, a standard sample differed from the others (p> 0.05), however, no significant difference for samples with addition of different concentrations of OF. Contributing to the increase in L* value in samples with fiber because the color of the OF (beige). TABLE 3: Parameters of color measurement of chicken nuggets added with different concentrations of Oat Fiber. L* a* b* Std 45.77

± 2.91 a 6.23

± 1.01 a 18.76

± 2.13 a 3% 49.61

± 2.47 b 6.26

± 0.91 a 21.84

± 2.44 c 4.5% 49.31

± 2.60 b 6.59

± 0.45 a 23.05

± 1.14 bc 6% 51.01

± 3.09 b 6.75

± 0.76 a 24.63

± 1.28 b Values in the same column with different letters indicate significant difference (p <0.05). Std – Standard Victorino (2008) [6] added wheat and oat fiber in emulsions with high levels of mechanically separated meat (CMS), and found that the results of L * are statistically significant (p ≤ 0.05), indicating that both adding wheat fiber and oat fiber influence linearly in the L * parameter, and the higher the addition of larger fibers higher the luminosity of the sample. In parameter b*, the standard sample differed statistically from other (p> 0.05). The formulation with the addition of 3% of OF had significant difference with the formulation of 6% of OF. And the formulation with addition of 4.5% was not statistically different from the other samples with fibers. Thus, it was found that the higher the addition of OF, the greater the intensity of the brown color. Table 4: Mean scores of sensory analysis by testing for multiple comparisons

Formulation Average Scores

3% 2.87 ± 1.26 a

4.5% 2.70 ± 1.22 a

6% 2.96 ± 1.08 a Means with the same letters in the same column indicate no difference at 5% by Tukey test. The results obtained in the sensory analysis, specifically for multiple comparisons can be

seen in Table 4, indicating that the three formulations are equal (p = 0.549) For the attributes of color, flavor and texture no significant difference (p <0.05) between samples of breaded chicken with addition of OF were observed. However, juiciness attribute the tasters noted difference between the samples at 5% significance. It was observed that a higher fiber addition, the higher the score given by the panel, being a score 1 the best and a score 5 worst value in the range of multiple comparison to a reference (R). Bortoluzzi (2009) [7] also found that products made with the highest concentration of orange fiber in bologna are often more dry. It can be observed from Figure 1 that formulation with addition of 3% of OF, presented the highest scores in all evaluated parameters, however, it was also not observed statistical difference among samples with added 4.5% and 6% of Oat Fiber (OF).

Chicken Nuggets with added Oat Fiber

Global Attributes

Color

Taste

Juiceness Texture

Figure 1 Average scores obtained by Multiple comparison for breaded chicken (nuggets) added with 3%, 4.5% and 6% of OAT FIBER.

IV. CONCLUSION

The data obtained in the development of this study indicate: - The chemical composition of the samples with addition of Oat Fiber met the criteria required by Brazilian legislation; - The formulation with addition of 6% Oat Fiber impact on the coloring of the final product, and interfered with the succulence of chicken nuggets; - All formulations showed good acceptability and indicated excellent purchase intent for formulations 3% and 4.5%.

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This work has proved to be possible develop formulations of restructured products with great acceptance, and showed good behavior with addition of OF. However, many variables must be evaluated if there is industrial scale production. An economic evaluation involving all areas must be performed in order to check availability of raw materials, target consumer, packaging and distribution system. These factors should be carefully evaluated to not impact the cost and sales of products, which could contribute to the success of the final product. ACKNOWLEDGEMENTS We are thankful to CAPES and UTFPR for supporting this work. FAGC and MRP CNPq research fellows. AG is CNPq/IC scholarship fellow and MAM and CAI are Fundação Araucária/IC scholarship fellows. REFERENCES [1] BRASIL. Ministério da Saúde. Agência Nacional de Vigilância Sanitária (Anvisa). Regulamento Técnico sobre Informação Nutricional Complementar. RDC, n°54, de 12 de novembro de 2012. Disponível em: http://portal.anvisa.gov.br/wps/wcm/connect/630a98804d7065b981f1e1c116238c3b/Resolucao+RDC+n.+54_2012.pdf?MOD=AJPERES. Acesso em 20 de janeiro de 2014. [2] AOAC. Official Methods of analysis of the assossiation analytical chemists. 18ed; Gaithersburg, Maryland, 2005. [3] SOARES, A. L. PSE (Pale, Soft, Exudative) em frangos: Implementação de parâmetro de cor e avaliação bioquímica e estrutural do filé (Pectoralis major). 2003. Tese (Doutorado em Ciência dos Alimentos) – Centro de Ciências Agrárias, Departamento de Ciência e Tecnologia dos Alimentos. Universidade Estadual de Londrina, Paraná. [4] ABNT. Associação brasileira de normas técnicas. Teste de comparação múltipla em análise sensorial de alimentos e bebidas: NBR 13.526. Rio de Janeiro, 1995. [5] BRASIL. Ministério da Agricultura Pecuária e Abastecimento. Instrução

Normativa n° 06 de 15 de fevereiro de 2001. Regulamento Técnico de Identidade e Qualidade de Empanados. Anexo III. Brasília, 2001. Disponível em: http://extranet.agricultura.gov.br/sislegis-consulta. Acesso em: 08 de novembro de 2012 [6] VICTORINO, L. C. S. Efeito da adição de fibras sobre as propriedades tecnológicas de emulsões com altos teores de carne de frango mecanicamente separada. 2008. Dissertação (Mestrado em Tecnologia de Alimentos) Universidade Estadual de Campinas, Campinas/SP. [7] BORTOLUZZI, R. C. Aplicação da fibra obtida da polpa da laranja na elaboração de mortadela de frango. 2009. Tese (Doutorado em Ciência dos Alimentos) Universidade de São Paulo. São Paulo/SP.

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http://extranet.agricultura.gov.br/sislegis-consulta

http://extranet.agricultura.gov.br/sislegis-consulta