APPLICATION OF ANTIBIOTICS IN

FOOD INDUSTRY

PRESENTED BY
DINESH KUMAR THAWAIT
 ANTIBIOTICS
Antibiotics are secondary metabolites produced by microorganisms
that inhibit or kill a wide spectrum of other microorganisms.
 Most of the useful ones are produced by molds and bacteria of the
genus Streptomyces.
 Some antibiotic like substances are produced by Bacillus spp., and at
least one, nisin, is produced by some strains of Lactococcus lactis.

 Three antibiotics have been investigated extensively as heat adjuncts

for canned foods:
 Subtilin,
 Tylosin, and
 Nisin.
 60 to 80 percent of all livestock and poultry will receive antibiotics in
feed or water or by systemic injection at some time during their
production lifespan.
 Preservatives Maximum Organisms Affected Foods
Tolerance
Ethyl formate 15-120 ppm Yeasts and molds Dried fruits, nuts
Caprylic acid — Molds Cheese wraps
Sodium nitrite 120 ppm Clostridia Meat-curing preparations
Dehydroacetic acid 65 ppm Insects Pesticide on strawberries,
Squash
Nisin 1% Lactics, clostridia Certain pasteurized cheese
spreads
Sodium diacetate 0.32% Molds Bread
Ethylene/ propylene 700 ppm Yeasts, molds, vermin Fumigant for spices, nuts
oxides
SO2/ sulfites 200-300 ppm Insects, microorganisms Molasses, dried fruits, wine
making, lemon juice (not to be
used in meats or other foods
recognized as sources of
thiamine)

Parabens 0.1 %f Yeasts and molds Bakery products, soft drinks,

pickles, salad dressings
Benzoic acid/ benzoates 0.1 % Yeasts and molds Margarine, pickle relishes, apple
cider, soft drinks, tomato
ketchup, salad dressings
Sorbic acid/ sorbates 0.2% Molds Hard cheeses, figs, syrups, salad
dressings, jellies, cakes
Propionic acid/ 0.32% Molds Bread, cakes, some cheeses,
propionates rope inhibitor in bread dough
 THE ANTIBIOTIC ISSUE
• Antibiotics are used in food animals for
– treatment or prevention of disease
– for increased production performance or increased
efficiency of use of feed consumed by the animal for
growth, product output, or
– modifying the nutrient composition of an animal product
– Enhance their growth and production performance first
demonstrated for poultry.
– fighting subclinical diseases and bolstering health defense
processes

• Various nutritional studies in chicks showed that

antibiotic-fermentation products influenced the
growth of chicks.
• By 1951, the addition of growth-promoting antibiotics
to feed had become standard practice.
 Some 15 considerations on the use of antibiotics as food
preservatives were noted by Ingram et al., and several of the key
ones are summarized as follows:

• The antibiotic agent should kill, not inhibit, the flora and
should ideally be destroyed on cooking for products that require
cooking.

• The antibiotic should not be inactivated by food components

or products of microbial metabolism.

• The antibiotic should not readily stimulate the appearance of

resistant strains.

• The antibiotic should not be used in foods if used

therapeutically or as an animal feed additive.
 Nisin
 Nisin was discovered in 1928 by Rogers and his workgroup.
 Nisin is a lantibiotic (contains the rare amino acids, meso-
lanthionine and 3-methyl-lanthionine instead of cysteine
and methionine), it is a bacteriocin.
 Nisin has approved bactericidal effect against most LAB; S.
aureus; L. monocytogenes, Bacillus and Clostridium
vegetative forms.
 Nisin ihibits synthesis of peptidoglycan: Inhibition of cell
wall synthesis
Nisin – E234
Nisaplin (Danisco)
GRAS status
1969 WHO and FDA
1983 EU
1988 US Food and Drug Agency
 The first food use of nisin was by Hurst to prevent the spoilage of Swiss
cheese by Clostridium butyricum.
 Most widely used compound for food preservation, with around 50
countries.
 Approved in 1988 for food use in the United States.
 The compound is effective against gram-positive bacteria, primarily spore
formers, and is ineffective against fungi and gram-negative bacteria.
Among some of its desirable properties as a food preservative are the
following:
• It is nontoxic.
• It is produced naturally by Lactococcus lactis strains.
• It is heat stable and has excellent storage stability.
• It is destroyed by digestive enzymes.
 Nisin as a heat adjunct in canned foods or as an inhibitor of heat-shocked
spores of Bacillus and Clostridium strains. Nisin, however, is used most widely
in cheeses.
 Usable levels are in the range of about 2.5-100 ppm.
Nisin is most often employed in dairy products—processed cheeses,
condensed milk, pasteurized milk, and so on. Some countries permit its use in
processed tomato products and canned fruits and vegetables. It is most stable
 Monensin

• Monensin, isolated from Streptomyces cinnamonensis.

• First described by Agtarap et al. in 1967.

• This antibiotic was approved by the FDA as a cattle feed additive in the
1970s, and it is used primarily to improve feed efficiency in ruminants.

• It inhibits gram-positive bacteria.

• Destroys selective permeability of cell membranes.

• Monensin is used extensively in the beef and dairy industries

 Natamycin

• It is a polyene antibiotic that is quite effective against yeasts and molds but not bacteria,
i.e, employed as a food fungistat.

• Natamycin, the name, as it was isolated from Streptomyces natalensis.

• Binds to ergosterol, a major component of the fungal cell membrane. When present in
sufficient concentrations, it forms pores in the membrane that lead to K+ leakage and death
of the fungus. Ergosterol is fairly unique to fungi, so the drug does not have such
catastrophic effects on animals.

• Its use is limited as a clinical agent, and it is not used as a feed additive.

• To control fungi on strawberries and raspberries, natamycin was compared with rimocidin
and nystatin, and it, along with rimocidin, was effective at levels of 10-20 ppm, whereas 50
ppm of nystatin were required for effectiveness.
 Tetracyclines
• Tetracyclines are both heat sensitive and storage labile in foods.
• Chlortetracycline (CTC) and oxytetracycline (OTC) were approved by the FDA in
1955 and 1956, respectively, at a level of 7 ppm to control bacterial spoilage in
uncooked refrigerated poultry.
• The efficacy of this group of antibiotics in extending the shelf life of refrigerated
foods was first established by Tarr and associates working with fish in Canada.
• Delay bacterial spoilage of fish and seafoods, poultry, red meats, vegetables,
raw milk, and other foods.
• CTC is generally generally more effective than OTC.
• When CTC is combined with sorbate, a natural organic compound to delay the
spoilage of fish, the combination has been shown to be effective for up to 14
days.
• Used to treat diseases in animals and are also used in feed supplements.
 Effects of Antibiotics on the Shelflife of Prepeeled Potatoes at
Room Temperature
E. A. M. ASSELBERGS, W. E. FERGUSON, AND W. P. MOHR,
Plant Research Institute, Research Branch, Canada Department of Agriculture, Ottawa,Canada
AND
K. F. MACQUEEN
Commercial Products Division, Atomic Energy of Canada, Ltd., Ottawa, Canada

• The quality of commercially prepeeled potatoes deteriorates rapidly during

nonrefrigerated transportation and storage.
• A major factor in this deterioration is the development of microorganisms.
• Ceponis and Friedman have shown that the shelflife of prepeeled potatoes at room
temperature is less than 24 hr.
• Similar results were reported by Feustel and Harrington.
• The application of antibiotics in prepacked vegetables has been reported to delay
bacterial spoilage by Carroll and co-workers.
• Recently, Francis et al. reported that the addition of oxytetracycline and
chlortetracycline to a sodium bisulphite dipping solution increased the shelflife of
prepeeled packaged potatoes.
 Subtilin
 Discovered and developed by scientists at the
Western Regional Laboratory of the USDA
 Its properties were described by Dimick et al.
 Produced by some strains of Bacillus subtilis.
 Effective against gram-positive bacteria.
 Subtilin is effective in canned foods at levels of 5-20
ppm in preventing the outgrowth of germinating
endospores.
 Used as a food preservative as it reduces the
thermal resistance of bacterial spores and so
permits a reduction in the processing time.
 Tylosin

• This antibiotic is a nonpolyene macrolide, as are the clinically

useful antibiotics erythromycin, oleandomycin, and others.
• It is more inhibitory than nisin or subtilin.
• Denny et al. were apparently the first to study its possible use in
canned foods.
• Used in animal feeds and also to treat some diseases of poultry.
• As a growth promotant in food producing animals.
• Used as a treatment of colitis, an inflammation of colon, in
small animals.
• Most effective against gram-positive bacteria.
• It inhibits protein synthesis by associating with the 50S
ribosomal subunit.
Fluoroquinolones in poultry
– Enrofloxacin
– Sarafloxacin

Quinolones inhibit the bacterial DNA gyrase enzyme, thereby inhibiting DNA
replication and transcription.
Campylobacter Resistance to Fluoroquinolones Increasing
• Most common food-borne infection in US
• 2.5 million case of diarrhea and 100 deaths per year
• 13% in 1998, 18% in 1999
• Fluoroquinolone use up 40% over same period

 Oct. 2000: FDA proposed to ban fluoroquinolones for therapeutic use in

poultry
 Major poultry producers -- Perdue, Tyson Foods, Foster Farms, ConAgra, Gold Kist,
Claxton, and Wayne Farms – say they have reduced or eliminated routine uses of
medically important antibiotics or the use of fluoroquinolones in sick birds
 Nitrofurans, particularly furazolidone (FZD), furaltadone (FTD), nitrofurantoin (NFT)
and nitrofurazone (NFZ), belong to a class of synthetic broad spectrum antibiotics .
 Nitrofurans were commonly employed as feed additives for growth promotion, and
mainly used for livestock (i.e. poultry, swine and cattle), aquaculture (i.e. fish and
shrimp) and bee colonies in the prophylactic and therapeutic treatment of bacterial
and protozoan infections such as gastrointestinal enteritis caused by Escherichia coli
and Salmonella spp., fowl cholera and coccidiosis black heads.
 Furazolidone was broadly used in European countries as an effective veterinary
antibiotic, especially in pig husbandry.
 In 1995, the use of nitrofurans for livestock production was completely prohibited
in the EU due to concerns about the carcinogenicity of the drug residues and their
potential harmful effects on human health.
 The use of nitrofurans for livestock has also been prohibited in countries such as
Australia, USA, Philippines, Thailand and Brazil.
 Where is the danger?
40% of the antibiotics used each year in the United States are for animals

Heavy use of antibiotics in factory farming of animals has caused

bacteria to develop antibiotic resistant strains.

Campylobacter bacteria – the most common cause

of food-borne illness in the US – increased its drug
resistance from 0% in 1991 to 20% in 1999.

The rate at which microorganisms in food-animal populations

become resistant to antibiotics is slow because of the short lifespans
and high turnover of these animal populations (Walton 1986).
Consequences of Agricultural Antibiotic Use

• Campylobacter fluoroquinolone resistance

• VREF (due to avoparcin use in chickens)

• MRSA in pork, chickens

• Gentamycin- and Cipro-resistant E. coli in chickens

 RESISTANCE TO ANTIBIOTIC DRUGS
• Use of drugs in food animals - Microorganisms can mutate to develop or acquire
resistance to antibiotic drugs.
• Is the microorganism zoonotic; that is, can it cause a human disease by moving
from the animal to a human, more virulent, treatable with other antibiotics?
• Passage of antibiotic-resistant bacteria from animals to humans occurs from
– direct contact with animals or their manure,
– through indirect exposure to food contaminated with animal-derived bacteria,
– from person-to-person contact after a primary exposure of nonfarm persons.
• The resistance of microorganisms arising from subtherapeutic use of penicillin,
tetracyclines, and sulfa drugs in agriculture is suggested by WHO (WHO 1997) to
be a high- priority issue.
• World Health Organization in 1997- “The use of any antimicrobial agent for
growth promotion in animals should be terminated if it is used in human
therapeutics or known to select for cross-resistance to antimicrobials used in
human medicine.”
• zoonotic pathogens:
– Salmonella DT-104 and
– Campylobacter jejuni
• Vancomycin and Ciprofloxacin resistance is linked to antibiotic use in
animals.
• In Europe there is strong evidence that one type of the vancomycin-
resistant enterococci (VRE - vanA) developed in animals fed an antibiotic
called avoparcin (a glycopeptide or vancomycin-like antibiotic).
• Vancomycin-resistance genes can spread from VRE to bacteria that are
much more common and aggressive such as the multi-resistant strains
of Staphylococcus aureus (MRSA).
• Use of enrofloxacin has resulted in the development of ciprofloxacin-
resistant strains of Salmonella spp and Campylobacter spp. These
resistant bacteria have subsequently caused human infections.
• When the glycopeptide, avoparcin, was used as a growth promoter in
food animals in Europe this resulted in the development and
amplification of vancomycin resistant enterococcus (VRE) and
subsequent colonisation by a significant percentage of the human
population via the food chain (between 2 and 17%).
 Antibiotic-Resistant Bacteria Have Been Transferred
from Animals to Humans (statistical data)

• Epidemiologic studies suggest transference of antibiotic-resistant E. coli and

campylobacter bacteria
• Examples:
– Chickens on farm fed tetracycline-supplemented feed and within 2 weeks 90% were
excreting all tetracycline-resistant E. coli. Within 6 months, 7 of 11 people living on
or near farm were excreting high numbers of resistant E coli.
– Percentage of Campylobacter jejuni resistant to quinolone in human isolates
increased from 0.8% in 1996 to ~3% in 1998. FDA approved the use of
fluoroquinolones in 1995.

• Epidemiologic studies with molecular subtyping show transference of antibiotic-resistant

campylobacter and salmonella bacteria
– Fluoroquinolone-resistant Campylobacter jejuni was isolated from 14% of 91 chicken
products obtained from retail markets in 1997.
– In 1998, salmonella bacteria resistant to ceftriaxone were isolated from a 12-year-old
boy who lived on a cattle farm.
• Enrofloxacin is administered to flocks of poultry through their water to control
mortality from E. coli and Pasteurella multocida organisms
• New evidence that enrofloxacin use in poultry has not been shown safe for humans
– FDA determined that the use of enrofloxacin in poultry causes the
development of a fluoroquinolone-resistant strain of campylobacter in poultry,
which, when transferred to humans, is a significant cause of fluoroquinolone-
resistant campylobacter infections in humans
The use of antibiotics in livestock production for increased feed efficiency is widespread. Such
use may indirectly access human food in the form of residuals in products such as meat and
milk. For years, scientists and health officials have warned of the risk of developing resistant
pathogens from feeding antibiotics to livestock. Recently, this concern was fueled by a fatal case
of salmonellosis caused by an antibiotic resistant strain linked to meat.
• Use of antibiotics in the treatment of mastitis has created problems for the milk processor
and consumer. Following treatment of mastitis with antibiotics, they may be found in the
milk in sufficient concentrations to inhibit dairy starter microorganisms and cause economic
losses to the cheese and fermented milk industries.

• Penicillin in very small concentrations found in milk may cause reactions in highly sensitive
individuals.

• When antibiotics are used to treat mastitis, dairymen should follow the prescribed
recommendations for withholding milk for human use following treatment.

• When adulterated milk leaves the farm, it is subjected to various processes in the milk plant.
Antibiotics in milk are relatively stable to pasteurization temperatures and above, as well as
to low temperatures (0–10° F.). Under refrigeration temperatures up to seven days of
storage, in raw and pasteurized milk there tends to be a loss in antibiotic activity.

• The presence of antibiotics in milk constitutes an adulteration under the Federal Food, Drug
and Cosmetic Act.
 Triphenyltetrazolium
Chloride as a Test for Antibiotic Substances i
nSummary
Milk

• A simple test for detection of inhibitory substances in milk is outlined.

– The test is based on the conversion of 2,3,5-triphenyltetrazolium chloride
(TTC) to 1,3,5-triphenylformazan.
– The reaction is characterized by a color change from the white to red in the
presence of growing bacterial cells.
– This conversion is inhibited by antibiotics at low levels of concentration.
– The total time for the test is 2½ hours.
 Findings on antibiotic resistance for food animals are as follows:
• Increases the risk of emergence of microorganisms that are resistant to
specific, and perhaps other, antibiotics.
• Resistance emergence should be classified with regard to each antibiotic
used, the concentration and dosage administered, the blood and tissue
concentrations attained, the bacterial species or strain affected, and the
animal species in which the drug is used.
• Zoonotic spread of pathogens to humans is very low. Postfarm food
processing, storage, and improper handling and cooking are major
contributors to the chain of events that allows the pathogen to
contaminate the product, proliferate on or in the food, and attain the
large numbers that cause disease.
• Three basic principles of antibiotic use were adopted
in the agriculture sector to reduce or eliminate
antibiotic resistance:
– Antibiotics that are ‘critical’ or ‘last-line’ for serious human
infections should not be used in food production animals
or agriculture.
– The use of antibiotics for prophylactic purposes in
animals should be kept to a minimum. The use of
methods (other than antibiotics) to prevent infections
should be expanded and developed.
– Antibiotics should not be used as growth promoters.
 REFERENCES:-
• James M. Jay, Modern Food Microbiology, Sixth Edition
• Foodborne Threats to Health: Policies, Practices, and Global Coordination
• Microorganisms in food, 2nd edition, Kluwer Academic / Plenum
Publishers, New York, Boston, Dordrecht, London, Moscow
• The Use of Drugs in Food Animals: Benefits and Risks (1999), Board on
Agriculture (BOA)
• Khem M. Shahani and , Paul J. Whalen, Department of Food Science and
Technology, University of Nebraska, Lincoln, Agricultural Uses of Antibiotics
• Journal of dairy sciences
• International journal of food microbiology
• Journal of food microbiology
• Agricultural uses of antibiotics, ACS publications