MICROBIAL FACTORS INFLUENCE FOOD CHEMISTRY AND QUALITYKatie Kirsch

Food Spoilage and Waste• According to the FAO

(2011)…• Approximately 1/3rd (1.3

billion tons) of food produced is lost or wasted annually

• Industrialized countries waste more than developing• Europe and North America:

95-115 kg/year (209-254 lb/year) per capita

• Sub-Saharan Africa and South/Southeast Asia: 6-11 kg/year (13-24 lb/year) per capita

Biogenic Amines (BA)• What are biogenic amines?

• Where are biogenic amines found?

• How are they produced?

• What health impacts do they have?

• What factors influence production?

• How can they be reduced/eliminated?

Amines• Organic, low molecular weight nitrogenous bases

• Eukaryotic cells• Essential precursors for hormones, alkaloids, nucleic acids and

proteins• Neurotransmitters

• Prokaryotic cells• Defense mechanisms• Generation of energy• Protection against acid

• Restore intracellular pH by consuming protons and excreting amines and CO2

Biogenic Amines

• Natural polyamines• Produced by plants, animals and

microorganisms• Important for nucleic acid regulation, protein

synthesis, and stabilization of membranes

• Biogenic amines• Enzymatic decarboxylation (removal of α-

carboxyl group) of free amino acids• Found in everything good

• Meat, cheese, chocolate, beer, wine, vegetables

Why do we care?• Health

• Migraines, headaches• Gastrointestinal problems• Toxicological consequences• Histamine poisoning• Cheese reaction• Secondary amines can form

carcinogenic nitrosamines in the presence of nitrites

• Food Quality• Off odors and flavors

?

Histamine Intoxication• Between 1998 and 2008 in the

United States (Gould et al., 2013)• 333 outbreaks • 1,383 illnesses

• Yellow fin, tuna, mackerel, mahimahi, bluefish

• Flushing, sweating, diarrhea, vomiting, headaches

• Onset of symptoms: 10 min-4h

Vickers and Safai, 2013

Nitrosamine Formation• Nitrosamines are highly carcinogenic

• Cadaverine

• P

BA FormationO

NH2OH

OH

Tyro sin e (T y r) Tyramine

H istid in e (H is)

O

NH2

N

NH

OH

Histamine

O

NH2NH

OH

T ryp toph an (T rp) Tryptamine

O

NH2

NH2 OH

Lysin e (L y s) Cadaverine𝑑𝑒𝑐𝑎𝑟𝑏𝑜𝑥𝑦𝑙𝑎𝑠𝑒→

𝑑𝑒𝑐𝑎𝑟𝑏𝑜𝑥𝑦𝑙𝑎𝑠𝑒→

𝑑𝑒𝑐𝑎𝑟𝑏𝑜𝑥𝑦𝑙𝑎𝑠𝑒→

𝑑𝑒𝑐𝑎𝑟𝑏𝑜𝑥𝑦𝑙𝑎𝑠𝑒→

BA Producing Microorganisms• Histamine

• Gram-negative• Hafnia alvei, Morganella morganii, Klebsiella pneumonia

• Gram-positive• Pediococcus damnosus, Leuconostoc spp., Lactobacillus curvatus

• Tyramine• Gram-positive

• Enterococcus, Lactobacillus, Leuconostoc, Lactococcus

• Phenylethlamine• Produced by tyrosine decarboxylation microorganisms

• Putrescine and cadaverine• Gram-negative

• Enterobacteriaceae (Citrobacter, Klebsiella, Escherichia, Proteus, Salmonella, Shigella), Psudomonadacea, Shewanellaceae

Factors Influencing BA Production• Quality of raw materials

• Microbial contamination• Processing conditions

• Storage conditions• Temperature-20-37 °C optimal• Time- general increase over time• pH- 2.5-6.5 optimal• Oxygen- strain specific• Presence of fermentable carbohydrate stimulates activity

• 0.5-2.0% optimal, >3% inhibitory glucose

• Water activity• Lower Aw, less microbial activity

Mean biogenic amine levels in foods (mg/kg)Product Histamine Tyramine Cadaverine Putrescine

Beer 1.4 6.1 1.3 - 1.5 3.3 - 3.5

Red wine 3.6 - 3.7 2.7 - 2.9 0.2 - 0.5 4.2 - 4.8

White wine 0.8-0.9 1.1 - 1.2 0.1 - 0.2 1.4 - 1.5

Dried anchovies 348 - - -

Fermented fish 7.7 - 11.4 47.2-49.1 14 - 17.3 13.4 – 17

Fermented sausage

23.0 - 23.6 136 37.4 - 38 84.2 - 84.6

Fresh cheese 3.2 - 38.5 12.8 - 48 10.7 - 45 5.5 - 41.3

Hard cheese 25.2 - 65.1 82.9 - 113 47.8 - 83.5 26.6 - 65.5

Fermented vegetables

39.4 - 42.6 45 - 47.4 26 - 35.4 264

European Food Safety Authority, 2011

Legal Limits for BA• FDA Guidance Level

• 50 mg/kg of histamine in scombroid or scombroid-like fish

• Slovak Republic• 20 mg/kg of histamine in beer• 200 mg/kg of histamine in fresh fish and fish products• 200 mg/kg of tyramine in cheese

• The European Union• <100 mg/kg of histamine in raw fish• <200 mg/kg of histamine in salted fish (Scombridae and Clupeidae

species)

Indicators of Food Quality• Useful for fresh and thermally treat products

• Little utility for fermented products

• BA levels generally increase with microbial load• Proposed limits for fresh meat (Hernandez-Jover et al.,

1996)

Quality BA concentration (mg/kg)

Good <5

Acceptable 5-20

Low 20-50

Spoiled >50

BA Control• BA formation can be controlled

by• Inhibiting microbial growth• Inhibiting decarboxylase activity

• Traditional• Temperature control• High-quality raw material• Good manufacturing practices• Amine-negative starter cultures

• Emerging• BA degrading enzymes and

cultures• Modified atmospheric packaging• Irradiation• High hydrostatic pressure• Food additives and preservatives

Huis in’t Veld, 1996

Reduction of BA by L. casei in Cheese (Herrero-Fresno, et al., 2012)

A) Monitoring of tyramine concentration in mini-cheeses made with E. durans 655. B) Monitoring of histamine concentration in mini-cheeses made with L. parabuchneri

QUESTIONS?

References• European Food Safety Authority. 2011. Scientific opinion on risk based control of

biogenic amine formation in fermented foods. EFSA Journal 9(10):2393.• Garcia-Ruiz, A., Gonzalez-Rompinelli, E.M., Bartolome, B., and Moreno-Arribas, M.V.

2011. Potential of wine-associated lactic acid bacteria to degrade biogenic amines. International Journal of Food Microbiology 148 (2011) p 115-120

• Gould, L.H., Walsh, K.A., Vieira, A.R., Herman, K., Wiliams, I.T., Hall, A.J., and Cole, D. 2013. Surveillance for foodborne disease outbreaks- United States, 1998-2008. Morbidity and Mortality Weekly Report 62(SS02), p 1-34.

• Hernandez-Jover, T., Izquierdo-Pulido, M., Veciana-Nogues, M.T., and Vidal-Carou, M.C. 1997. Effect of starter cultures on biogenic amine formation during fermented sausage production. Journal of Food Protection 60 p 825-830

• Herrero-Fresno, A., Martinez, N., Sanchez-Llana, E., Diaz, M., Fernandez, M., Martin, M.C., Ladero, V., Alvarez, M.A. 2012. Lactobacillus casei strains isolated from cheese to reduce biogenic amine accumulation in an experimental model. International Journal of Food Microbiology 157(2) p297-304.

• Huis in’t Veld, J.H.J. 1996. Microbial and biochemical spoilage of foods: an overview. International Journal of Food Microbiology 33, p 1-18.

• Vickers, J., and Safai, B. 2013. Scombroid Poisoning. New England Journal of Medicine 368(31).