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ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD PRODUCTS Antanas Šarkinas Food institute of Kaunas University of Technology, Taikos pr. 92, LT-51180, Kaunas; [email protected]

ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; [email protected]. Spices

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Page 1: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

ANTIBACTERIAL ACTIVITY OF PLANT

EXTRACTS IN FOOD PRODUCTS

Antanas Šarkinas

Food institute of Kaunas University of Technology, Taikos pr. 92, LT-51180,

Kaunas; [email protected]

Page 2: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Spices

• Spices have been widely consumed throughout history in the human diet not only as flavouring substances but also as antimicrobial agents.

• Numerous reports demonstrate an inhibitory effect of essential oils and extracts, as well as purified compounds isolated from various plants on the growth of microorganisms; such substances have been used for preserving foods and drinks.

Page 3: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Effect of spices in food preservation

• Since the very first scientific experiments on the antimicrobial properties of spices, herbs and their components which were used in the end of the 19th century the interest in this topic has not diminished. Hoffman and Evans (1911) were the first who performed a laboratory study on the effect of spices in food preservation. They found that cinnamon, mustard and clove were useful in preserving apple juice.

Page 4: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Antimicrobial test

• The assessment of antimicrobial activity wasperformed on

• Gram-positive Listeria monocytogenes (ATCC 9117), Bacillus cereus (ATCC 10876), Bacillussubtilis (ATCC 6633), M. luteus (ATCC 9341), S. aureus (ATCC 25923),

• and Gram-negative E. coli (ATCC25922), Enterobacter aerogenes (ATCC 13048), Salmonella typhimurium (ATCC 14028), andSalmonella agona, Enterococcus faecalis (ATCC 29212), bacterial test cultures.

Page 5: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Strains of yeast

• In addition, 8 strains of yeast were used: Debaryomyces hansenii, Trichosporoncutaneum, Kluyveromyces marxianus var. lactis, Saccharomyces cerevisiae, Candidaparapsilosis, Torulaspora delbrueckii, Pichiakluyveri, and Rhodotorula rubra.

• They were isolated from dairy products, equipment washing liquid, and the air ofindustrial premises.

Page 6: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Agar well diffusion method

• The antimicrobial properties were evaluated by the agarwell diffusion method.

• Bacteria were grown in peptone-soy bouillon (LAB 04, LABM) for 24 h at 37 C.

• After cultivation, test culture cells were mixed using a minishaker MS 1 (Wilmington, N.C., U.S.A.) and the cellsuspensions were adjusted according to McFarland nr0.5 standard (Hood and others 2003). The suspension ofbacteria cells was introduced into the dissolved mediaand cooled to 47 C;

• 10 ml was pipetted into a 90-mm diameter Petri plate. Eight-mm diameter wells were punched in the agar andfilled with 50 µL of extracts.

• The plates were incubated overnight at 37 C.

Page 7: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Yeasts

• Yeasts were grown on a potato dextrose agar slant (LAB 98, LAB M) for 48 h at 25 C.

• After cultivation, the yeast cells were washed with saline and the cell suspensions were treated as previouslymentioned.

• The media containing the yeast cells and oils wasincubated overnight at 25 C.

• A 50 µL volume of extract was added to the agar wellsusing an automatic pipettor.

• After incubation, the inhibition zones were measuredwith calipers to an accuracy of 0.1 mm and the effectwas calculated as a mean of 3 replicate tests.

Page 8: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Press vial into agar

Page 9: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Agar well diffusion method

Page 10: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Microbiological tests

• Microbiological tests showed that extract from coriander, celery, garlic, marjoram inhibit the growth of test cultures. Effectiveness of celery extract was higher in comparison with other extracts.

• The results showed that grampositivebacteria were more sensitive to extracts of celery, parsley, coriander, tarragon and lemon balm than gramnegative bacteria.

Page 11: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Preservative effect

• The possibility to stabilize a number of coliformsin meat mince was also evaluated. The extracts of plants did not have significant influence on the number of coliforms during cold storage.

• The test with ginger showed that it was contaminated with microorganisms which increased total number of bacteria in mince, and it did not have any preservative effect during storage.

Page 12: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Preservative effect

• Caraway extract inhibited growth of all test cultures, but its influence was not strong, zones of inhibition were not wide. Differences between sensitivity of salmonella were determined. S. typhimurium and S. agona had smallest sensitivity to caraway extract.

• Count of bacteria and coliform bacteria had growth in all samples with coriander or caraway extracts. In case with higher amount of extracts growth rate was slower.

Page 13: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Acceptability

• Consumer focus group had evaluated acceptability of cooked minced meat samples with plant extracts. Samples with 0,2 % of coriander extracts were evaluated as most acceptable but samples with 1 % were not acceptable for consumer at all.

• Consumer test results showed that increase in concentration of coriander extract in meat gives lower acceptability of samples.

Page 14: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Antimicrobial properties ofcranberry extracts

• Investigation of the antimicrobialproperties of cranberry extracts by thediffusion to agar method showed that B. cereus and M. luteus were the mostsensitive (average zones of inhibition were

• 2.28 and 2.24 cm),

• L. monocytogenes was found to haveaverage resistance, and E. coli was theleast sensitive

Page 15: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Different cultivars

• A comparison of results from the different cultivarsshowed that the largest zones of inhibition (1.70 to 2.55 cm) weremade by the extract of the cultivar “Pilgrim.”

• The cultivar “Stevens” had the 2nd highest antimicrobial

• properties.

• Extracts of less active “Ben Lear” and “Black Veil”showed zones of 1.60 to 2.05 cm in diameter.

• All investigated test cultures were more sensitive to theceftazidime/clavulanic acid 30/10 µg sensi-disc, exceptB. cereus, which was more sensitive to the extracts fromberry and berry cakes

Page 16: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Natural antimicrobial agents

• The differences between antimicrobial activity ofberry and berry cake extracts were notsignificant.

• Antimicrobial berry compounds may have usefulapplications as natural antimicrobial agents forthe food industry as well as for the medicalpurposes

• Cranberry extracts did not have any effect on thegrowth of any of the 8 yeast species studied.

Page 17: ANTIBACTERIAL ACTIVITY OF PLANT EXTRACTS IN FOOD … · Antanas Šarkinas Food institute of KaunasUniversity of Technology, Taikos pr. 92, LT-51180, Kaunas; direktorius@lmai.lt. Spices

Antimicrobialproperties of cranberry extracts

0 5 10 15 20

E. coli

S. aureus

S. typhimurium

E. faecalis

B. subtilis veg. ląstelės

B. subtilis sporos

L. monocytogenes

Bak

terij

ų ku

ltūro

s

Slopinimo zonos skersmuo, mm

Paprastoji spanguolė Stambiauogė spanguolė