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Antibiotic Residue- Food Producing Animal Origin and Its Impact on Human Health

Ramesh K. Nirala K. Anjana K. G. Mandal C. Jayachandran
Vol 8(10), 61-69

Antibiotics are mainly employed for chemotherapeutic, prophylactic purposes and also used as feed additives to promote growth and improve feed efficiency. However, antibiotic residues in animal products may occur, when administration of drug in extra label fashion and not following of withholding or withdrawal period after treatment. A withdrawal period is established to safeguard human from exposure of antibiotic added food. The withdrawal time is the time required for the residue of toxicological concern to reach safe concentration as defined by tolerance. Many of the administered drugs are not completely absorbed from gut and excreted through faeces and urine as either parent compound or its toxic metabolites. The application of manure or farm effluents in agricultural land leads to selection of resistant bacteria, development and transmission of antibiotic resistance genes in the microbes. The antibiotic resistance in animal and human leads to poor response to treatment during illness. The antibiotic residues in animal product causes harmful effect on health and also interfere with the processing of milk and milk products. The present review focuses on antibiotic residues in animal products and its impact on environments and human health.

Keywords : Antibiotic Buffalo Cattle Milk Residue Withdrawal Period

Antibiotics are substances either produced naturally by living organisms or synthetically in the laboratory, and they are able to kill or inhibit the growth of microorganisms. Antibiotics residue in foods of animal origin are one of the sources of concern among the public and medical health professionals (Bren, L. et al., 2002 ) Antibiotics are also used as feed additives for the purpose of livestock health maintenance. Apart from this, they are also used as feed additives as growth promoters may result in presence of antimicrobial residues in milk and dairy products, and can contribute to the development of microbial drug resistance and the spread of resistant bacteria (Stolker, A. A. M. et al., 2007). The residues of Veterinary Medicinal Product i.e. antibiotics present a potential risk to the human population, particularly with the appearance of allergic reactions and interferences of intestinal micro-flora (Dewdeny, J. M. et al., 1991). From the technological prospective, residues of antimicrobial agents in milk can cause significant losses in fermented products, by inhibition of bacterial fermentation in the production processes of cheese or yoghourt (Molina, M. P. et al., 2003). Therefore, accurate detection of low levels of antimicrobial drug residues in milk is of great importance for the dairy industry and for farmers, with a purpose to ensure that the contaminated milk from individual cows are not consigned to the bulk tank (Mitchell, J. M. et al., 2002).The fluoroquinolones antibiotics have wide range of  antibacterial activity and have seen increasing use in veterinary medicine because of their effectiveness in treating bacterial infection (A. L. Cinquina et al., 2003). Antibiotic residues in feedstuffs are currently a problem of some magnitude in different parts of the world, particularly due to associated public health concerns that include hypersensitivity reactions, antibiotic resistance, toxicity, teratogenicity, and carcinogenicity (Wageh Sobhy Darwish, et al., 2013). Use of Antibiotic that might result in deposition of residues in meat, milk and eggs must not be permitted in food intended for human consumption. If use of antibiotics is necessary as in prevention and treatment of animal diseases, a withholding period must be observed until the residues are negligible or no longer detected. Hence, the present review focuses on antibiotic residues in animal products and its impact on environments and human health.

Application of Antibiotics

The antibiotics nowadays used for improved performance in growth especially in broilers and fatteners. They may produce improved growth rate because of thinning of mucous membrane of the gut, facilitating better absorption, altering gut motility to enhance better assimilation, producing favorable conditions to beneficial microbes in the gut of animal by destroying harmful bacteria and partitioning proteins to muscle accretion by suppressing monokines. Antibiotics also favour growth by decreasing degree of activity of the immune system, reduced waste of nutrients and reduce toxin formation. In most of the cases only young growing animals and poultry are responsive to antibiotic mediated growth promotion. Indiscriminating use of antibiotics in all cases of pyrexia, inflammation, wounds and viral diseases have widespread residual effects on edible tissues. The use of antibiotics only in specific conditions is justified because the roll of microbial agents is mainly to kill the rapidly dividing invading cells. Animals and poultry are receiving sub therapeutic levels of antibiotics to prevent possible infection. But the antibiotics are specific to their spectrum of activity only in the active multiplying stage of bacteria. But it will not provide overall protection. Only in certain cases like dry cow therapy and surgical procedures are wanting of antibiotic prophylaxis. Antimicrobials are used either directly or indirectly during the production processing and storage of milk and milk products. Direct contamination of milk may occur from air and water during processing, storage and transportation. Besides feed given to animals is also source of indirect contamination. Man will be the ultimate consumer of these antibiotic residues. There are some causes of miscellaneous use like lack of awareness, lack of extension activities, inadequate literature supplied by manufacturers, lack of safer drugs and exploitation of more production and profit from animals. FDA prohibits the extra label use of chloramphenicol, furazolidone, nitrofurazone, sulphonamide drugs, and flouroquinolones in lactating animals.

Antibiotic Residues

Several antibiotic classes are extensively administered to food-producing animals, including tetracyclines, sulfonamides, fluoroquinolones, macrolides, lincosamides, aminoglycosides, beta-lactams, cephalosporins and others (Woodward, K. 2004; Jank, L. et al., 2017).

Table 1: Maximum Residues Limit (MRL) of antibiotic as per Codex Alimentarius commission

S. No. Name of Antibiotics Substrate  Maximum Residual Limit  (µg/kg)
1 Benzyl penicillin Milk 4
Muscle/liver 50
2 Ampicillin Milk 4
3 Amoxycillin Milk 4
Muscle/liver 50
4 Oxacillin Milk 30
5 Cloxacillin Milk 30
6 Dicloxacillin Milk 30
7 Tetracycline Milk 100
Muscle 200
8 Oxytetracycline Milk 100
Muscle 200
9 Chlortetracycline Milk 100
Muscle 200
10 Streptomycin Liver-Poultry 200
11 Dihydrostreptomycine Milk 200
Muscle 600
12 Gentamycine Milk 200
Muscle 100
13 Sulphonamides Milk 100
14 Ceftiofur Milk 100
Muscle 1000

Almost 90% of all antibiotics used in farm animals and poultry are reported to be administered at sub-therapeutic concentrations. About 70% of this is for the purpose of disease prevention and 30% are for growth promotion (Kebede, G. et al., 2014). The risk of residue from the milk is higher in developing countries compared to develop one. This might be related with lack of facilities for detection and regulatory bodies that control the drug residues level in foods in the form of maximum residue limits (Kebede, G. et al., 2014). The MRL is defined as the maximum concentration of a residue, resulting from the registered use of an agricultural or veterinary chemical that is recommended to be legally permitted or recognized as acceptable in or on a food, agricultural commodity, or animal feed.

Screening/Analytical Method

Microbial Inhibition Test

It is a qualitative or semi-quantitative test is used to detect antibiotic residues in milk and meat. Muller Hinton or Nutrient agar is commonly used to perform agar diffusion test. These tests comprise spores of specific bacteria (Bacillus subtilis, Bacillus stearothermphillus, Micrococcus luteus etc), sensitive to particular antibiotics on agar gel including nutrients for bacterial growth and a pH indicator (Chafer-Pericas C. et al., 2010). After addition of milk, the plate is incubated at appropriate temperature for growth and germination of bacteria. In the absence of antibiotic residues, the growth of bacteria can be detected visually either by the change of opacity of the agar medium or by the colour change of the pH indicator ( Hakem, A. et al., 2013, Navratilova P. et al., 2008, Dang, P.K. et al., 2010). This method is cheap and simple, showed 100% sensitivity and specificity for screening of residues (Dang, P. K. et al., 2010), 85.51% of poultry meat samples were positive for antibiotic residue, most of them contained β-lactams and tetracyclines (75.81%), macrolides (44.35%), sulphonamides (36.29%), aminoglycosides (13.71%) group (Thompkinson, K., 2012) and macrolides (12.5%) group of antibiotics in commercial eggs of urmia, Iran (Ehsani, A. et al., 2015).


Biosensor is a latest technique used for screening of antibiotics residues in milk. The sensitivity and selectivity of biosensors are comparable to immunoassay methods (Rinken, T. et al., 2006). These sensors have shown successful detection of β-lactams (β-Ls), tetracycline, streptogramin and macrolide antibiotics at nanogram per millilitre concentrations in milk and serum (Weber, C. C. et al., 2005, Toldra, F. et al., 2006). Microbial biosensor is mostly used for detection of quinolones (Qs) and tetracyclines (TCs). However, these were not sensitive toward detection of macrolides, β-Ls, aminoglycosides and sulphonamides (Kivirand, K. et al., 2015).

Enzyme Linked Immunosorbent Assay (ELISA)

ELISA is most useful and specific test for screening of drug residues in meat, milk and egg. The Competitive ELISA is commonly used for quantitative analysis of tetracycline, fluoroquinolones and chloramphenicol in meat (Yibar, A. et al., 2011, Kim, D. P. et al., 2013). Competitive indirect chemiluminescent enzyme-linked immunoassay (CL-ELISA) is used to determine the chloramphenicol (CAP) residues in milk and chicken muscle. The sensitivity of CL-ELISA is 2-3 times higher than conventional ELISA (Tao, X. et al., 2012) and able to detect trace amount of CAP as low as 3.19 ng/kg in chicken muscle. In India (Punjab), Gaurav et al., 2014 reported that tetracycline residues in milk by competitive ELISA. Out of 133 cattle milk sample, 18 samples were found to be contaminated with tetracycline. The concentration of tetracycline residues in cattle milk samples were found to be in the range 16-134.5 μg/l. Three samples exceeded the maximum recommended tetracycline antibiotic residue levels (MRLs). Sultan et al., 2014 reported that enrofloxacin residues in liver sample of poultry, sheep and cattle collected from slaughter house of Mousl city, Iraq. Out 30 samples from each species 17 poultry sample, 8 cattle sample and 5 sheep sample exceeded the maximum residue limits (Permitted MRL by European agency 100-300 μg/kg). The concentration of enrofloxacin in liver sample of poultry, cattle and sheep were 10-10690, 30-3610 and 20-1320 μg/kg respectively. This may be heavy use of enrofloxacin in poultry industry to control diseases.

High Performance Liquid Chromatography (HPLC)

Currently, HPLC is one of the most powerful tools in analytical chemistry. HPLC usage is increasing day by day in the field of residue analysis because, these having variety of mobile phases, the extensive library of column packings and the variation in modes of operations (Jank, L. et al., 2017). The residue analysis of oxytetracycline and penicillin G in milk collected from Nazareth dairy farms, Ethopia (Senyuva et al., 2000). Out of 400 milk samples 48 samples were found oxytetracycline and penicillin G in the range of 45-192 and 0-28 μg/l, respectively. A total of 497 raw milk samples were collected from different region of individual farms at Macedonia and analysed for chloramphenicol, sulfonamides, quinolones and tetracyclines by HPLC-Diode array detection. The concentration of sulphonamides, quinolones and tetracycline in the range of 13.5-147.9, 0.6-22.0 and 17.4-149.1μg/kg, respectively. None of the samples showed chloramphenicol above MRL (Elizabeta et al., 2011). A total number of 250 milk samples consisting of 125 from organized and 125 from unorganized sectors were randomly collected out of which 50 sample from each districts of Bihar. It was found that out of all the milk samples analyzed BY HPLC , eight samples (3.2 %) were found to contain antibiotics residue. Three samples (1.2 %) exceeded the maximum residue levels (MRL) for enrofloxacin antibiotics according to the regulation set by the European Union and the Codex Alimentarious Comission.(Nirala, R. K. et al., 2017). In Iran cattle tissue (Triceps muscle, gluteal muscle, diaphargam, kideney and liver) from local market were examined for tetracycline group of antibiotics (tetracycline, oxytetracycline and chlortetracycline) by HPLC method. The tetracycline concentration in Triceps muscle, gluteal muscle, diaphargam, kideney and liver were 176.3, 405.3, 96.8, 672.4 and 651.3 ng/g, respectively. The concentrations of tetracyclines were higher in liver and kidney sample compared to other sample (Abbasi, M. M. et al., 2012) and it was higher in cured meat product (Senyuva et al., 2000).

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Nowadays, the most frequently used analytical tool for detection of a large number of multiclass veterinary drug residues in food (Martins, M.T. et al., 2014, Layada, S. et al., 2016 ). The analysis of antibiotics in milk by LC-MS/MS is more specific and more reliable. Simultaneous determination of 14 antibiotics from different classes includes five β-lactams, four sulfonamides, three tetracyclines, one macrolide and one cephalosporin by liquid chromatography with electrospray ionization (LC-ESI) and triple quadrupole mass spectrometry (MS/MS) ( Martins-Junior HA et al., 2007), enrofloxacin and tetracycline residues in chicken and pork by LC-MS/MS (Kim, D.P. et al., 2013). Aminoglycoside in animal tissue includes bovine kidney, ovine kidney and poultry liver (Plozza, T. et al., 2011). Out of seventy two samples, twelve samples showed aminoglycosides includes neomycin, streptomycin and dihydrostreptomycin exceeded the MRL. The concentration of neomycin, streptomycin and dihydrostreptomycin were 10000, 300 and 300 μg (Plozza, T. et al., 2011).

Table 2: Antibiotic residue in food producing animal origin

S. No. Name of antibiotics Instrumentation  Substrate  Residual level(ppb) References 
1 Tetracycline HPLC-DAD Milk 17.4 – 149.1 Elizabeta et al., 2011
2 Tetracycline ELISA Milk 16-135.5 Gaurav et al., 2014
3 Gentamicin ELISA Milk 42-360 Senyuva et al., 2000
4 Streptomycin ELISA Milk 90 Zeina et al., 2013
5 Penicillin HPLC Milk 0-28 Abebew et al., 2014
6 Sulfamethoxazole UPLC-MS/MS Milk 4.2 Han et al., 2015
7 Sulphonamide HPLC-DAD Milk 13.5-147.9 Elizabeta et al., 2011
8 β-lactams, LC-MS/MS Milk Martin-Junior et al., 2007
Macrolides and
9 Quinolone HPLC-DAD Milk 0.6-22.00 Elizabeta et al., 2011
10 Enrofloxacin & Tetracycline ELISA Liver-Poultry 10-10690 Sultan et al., 2014

Preventive Measures to Minimizing the Antibiotic Residue in Livestock Products

This section includes recommendations and measures to reduce antibiotic residues in foods of animal origin-

  1. Reduce antibiotics use in food animal rearing. Many developed countries have banned its use as growth promoters.
  2. Rapid screening methods should be developed for detecting and segregating samples contains above MRL levels of antibiotics.
  3. Appropriate MRLs need to be set by regulatory bodies and should enforce it.
  4. Appropriate withdrawal periods should be strictly followed and enforced to make the meat rendered safe for human consumption.
  5. Improve the individual and organizational aware by enhancing proper knowledge dissemination.
  6. Follow best hygiene practices during animal rearing and avoid unwanted use of antibiotics.
  7. Alternates to antibiotics like bio control measures and Ethno-veterinary practices should be developed and followed.
  8. Organic poultry farming may be encouraged by providing appropriate incentives to the farmers in form of subsidies.
  9. Use of proper processing techniques to inactivate the antibiotic residue, e.g. refrigeration causes inactivation of penicillin.
  10. Use of activated charcoal, resins and UV irradiation to inactivate residues.


Use of antibiotics as feed additives at sub therapeutic dose should be strictly prohibited. For therapeutic purpose, it must be used in proper dose for proper time. There are two major concerns in the presence of antibiotic residues in milk, meat and egg. One is allergic reaction even at smaller dose, another development of antibiotic resistance and disruption of soil microbial community. Monitoring of antibiotic residues in milk and milk products, meat and meat products, egg, faeces and urine is necessary to safeguard the health of the consumers as well as minimize environmental contamination.


The authors are thankful to Indian Council of Agricultural Research, New Delhi and   Associate Dean-cum-Principal, Bihar Veterinary College, Patna-800014, Bihar State, India for providing funding along with   necessary facilities to conduct the present study.


  1. A.L. Cinquina et al. (2003): Determination of enrofloxacin and its metabolite ciprofloxacin in goat milk by high-performance liquid chromatography with diode-array detection Optimization and validation Journal of Chromatography. A 987, 221–226.
  2. Abbasi MM, Nemati M, Babaei H, Ansarin M, Nourdadgar AOS.(2012): Solid-Phase extraction and simultaneous determination of tetracycline residues in edible cattle tissues using an HPLC-FL method. Iranian Journal of Pharmaceutical Research. 2012; 11(3):781- 787.
  3. Bren, L. (2002): The Battle of the Bugs: Fighting Antibiotic Resistance. USDA consumer magazine; July-Aug. http//
  4. Chafer-Pericas C, Maquieira A, Puchades R (2010): Fast screening methods to detect antibiotic residues in food samples. Trends in Analytical Chemistry. 29(9):1038-1049.
  5. Dang PK, Degand G, Danyi S, Pierret G, Delahaut P, Ton VD et al.(2010): Validation of a two-plate microbiological method for screening antibiotic residues in shrimp tissue. Analytica Chimica Acta. 2010; 672:30-39.
  6. Dewdeny, J. M., Maes, L., Raynaud, J.P., Blanc, F., Scheid, J.P., Jackson, T., Lens, S., Verschueren, C. (1991): Risk assessment of antibiotic residues of beta-lactams and macrolides in food-products with regard to their immunoallergic potential, Food and Chemical Toxicology 29, 477-483
  7. Ehsani A, Hashemi M (2015): Determination of antibacterial drug residues in commercial eggs distributed in Urmia, Iran. Journal of Food Quality and Hazards Control. 2:61-65.
  8. Elizabeta DS, Zehra HM, Biljana SD, Pavle S, Risto U. (2011): Screening of veterinary drug residues in milk from individual farms in Macedonia. Macedonian Veterinary Review. 34(1):5-13.
  9. Gaurav A, Gill JPS, Aulakh RS, Bedi JS. (2014): ELISA based monitoring and analysis of tetracycline residues in cattle milk in various districts of Punjab. Veterinary World. 7:26-29.
  10. Hakem A, Titouche Y, Houali K, Yabrir B, Malki O, Chenouf N et al. (2013): Screening of antibiotics residues in poultry meat by microbiological methods. Bulletin of University of Agricultural Sciences and Veterinary Medicine. 70(1):77-82.
  11. Han RW, Zheng N, Yu ZN, Wang J, Xu XM, Qu XY et al.(2015): Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS. Food Chemistry. 181:119-126.
  12. Jank L, Martins MT, Arsand JB, Motta TMC, Feij TC, Castilhos TDS et al. (2017): Liquid chromatography-tandem mass spectrometry multiclass method for 46 antibiotics residues in milk and meat: Development and validation. Food Analytical Methods. 1-13.
  13. Jank L, Martins MT, Arsand JB, Motta TMC, Feij TC, Castilhos TDS et al. (2017): Liquid chromatography-tandem mass spectrometry multiclass method for 46 antibiotics residues in milk and meat: Development and validation. Food Analytical Methods.  1-13.
  14. Kebede G, Zenebe T, Disassa H, Tolosa T.(2014): Review on detection of antimicrobial residues in raw bulk milk in dairy farms. African Journal of Basic & Applied Sciences. 6(4):87-97.
  15. Kim DP, Degand G, Douny C, Pierret G, Delahaut P, Ton VD et al. (2013): Preliminary evaluation of antimicrobial residue levels in marketed pork and chicken meat in the red river delta region of Vietnam. Food and Public Health. 3(6):267-276.
  16. Kivirand K, Kagan M, Rinken T. (2015): Biosensors for the detection of antibiotic residues in milk. Published in: Biosensors – Micro and Nanoscale Applications, Intech. 425-456.
  17. Layada S, Benouareth DE, Coucke, Andjelkovic M. (2016): Assessment of antibiotic residues in commercial and farm milk collected in the region of Guelma (Algeria). International Journal of Food Contamination. 19(3):1-16.
  18. Martins MT, Melo J, Barreto F, Hoff RB, Jank L, Bittencourt MS et al. (2014): A simple, fast and cheap non-SPE screening method for antibacterial residue analysis in milk and liver using liquid chromatography–tandem mass spectrometry. Talanta. 129:374-383.
  19. Martins-Junior HA, Tereza A, Kussumi, Alexandre Y, Wang, Lebre DT. (2007): A Rapid method to determine antibiotic residues in milk using liquid chromatography coupled to electrospray tandem mass spectrometry. Journal of the Brazilian Chemical Society. 18(2):397-405.
  20. Mitchell, J.M., Groffi ths, M.W., Mcewen, S.A., McNab, W.B., Yee, A.J. (2002): Antimicrobial drug residues in milk and meat: Causes, concerns, prevalence, regulations, tests and test performance, Journal of Food Protection 61, 742-756.
  21. Molina, M.P., Althausb, R.L., Molinac, A., Fernándeza, N. (2003): Antimicrobial agent detection in ewe’s milk by the microbial inhibitor test brilliant black reduction test – BRT AiM®, International Diary Journal 13, 821-826.
  22. Navratilova P. (2008): Screening methods used for the detection of veterinary drug residues in raw cow milk- A review. Czechoslovakian Journal of Food Science.  26(6):393-401.
  23. Plozza T, Trenerry VC, Zeglinski P, Nguyen H, Johnstone P.(2011): The confirmation and quantification of selected aminoglycoside residues in animal tissue and bovine milk by liquid chromatography tandem mass spectrometry. International Food Research Journal. 18(3):1077-1084.
  24. Rinken T, Riik H. (2006): Determination of antibiotic residues and their interaction in milk with lactate biosensor. Journal of Biochemical and Biophysical Methods. 66:13-21.
  25. Senyuva H, Ozden T, Sarica DY. (2000): High-performance liquid chromatographic determination of Oxytetracycline residue in cured meat products. Turkish Journal of Chemistry. 24:395-400.
  26. Stolker, A.A.M., Zuidema, T., Nielen, M.W.F. (2007): Residue analysis of veterinary drugs and growth-promoting agents. Trends in Analytical Chemistry 26, 967-979.
  27. Sultan IA. (2014): Detection of Enrofloxacin in livers of livestock animals obtained from a slaughterhouse in Mosul City. Journal of Veterinary Science and Technology. 5(2):1-3.
  28. Swatantra S, Shukla, Tandia N, Nitesh K, Paliwal R. (2014): Antibiotic Residues: A global challenge. Pharma Science Monitor. 5(3):184-197.
  29. Tao X, Jiang H, Zhu J, Niu L, Wu X, Shi W et al. (2012): Detection of ultratrace chloramphenicol residues in milk and chicken muscle samples using a Chemiluminescent ELISA. Analytical Letters. 45(10):1254-1263.
  30. Thompkinson K. (2012): Quality assessment of milk and milk products, New India publishing agency, New Delhi, India. 226-227.
  31. Toldra F, Reig M. (2006): Methods for rapid detection of chemical and veterinary drug residues in animal foods. Trends in Food Science and Technology. 2006; 17:482- 489.
  32. Weber CC, Link N, Fux C, Zisch AH, Weber W, Fussenegger M.(2005): Broad-spectrum protein biosensors for class specific detection of antibiotics. Biotechnology and Bioengineering. 89:9-17.
  33. Woodward K. (2004): The evolution of safety assessments for veterinary medicinal products the European Union. Veterinary Human Toxicology: 46:199-205.
  34. Yibar A, Cetinkaya F, Soyutemiz GE (2011): ELISA screening and liquid chromatography-tandem mass spectrometry confirmation of Chloramphenicol residues in chicken muscle, and the validation of a confirmatory method by liquid chromatography-tandem mass spectrometry. Poultry Science. 90:2619-2626.
  35. Zeina K, Pamela AK, Fawwak S. (2013): Quantification of antibiotic residues and determination of antimicrobial resistance profiles of microorganisms isolated from bovine milk in Lebanon. Food and Nutrition Sciences. 4:1-9.
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