ANALYSIS OF MICROBIOLOGICAL QUALITY AND ANTIBIOTIC RESISTANCE PATTERNS IN MILK SUPPLY CHAIN

MICROBIOLOGICAL QUALITY AND ANTIBIOTIC RESISTANCE PATTERNS IN MILK

Authors

  • Farhana Rinky Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh
  • Sompa Reza Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh
  • Abira Nowar Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh
  • Supriya Ghosh Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh
  • Asma Rahman Food Nutrition and Agriculture Research Division, Centre for Advanced Research in Sciences (CARS), University of Dhaka, Dhaka, Bangladesh
  • Sharmin Rumi Alim Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh

DOI:

https://doi.org/10.3329/brc.v10i1.70686

Keywords:

Milk Supply Chain, Multi-antibiotics Resistance, Virulence Genes, Residual Antibiotics

Abstract

The widespread consumption of milk for its nutritional value and health benefits brings the risk of milk-borne diseases due to the presence of various microorganisms, including antibiotic-resistant pathogens. This has led to an increased focus on ensuring the safety of milk products across the supply chain by dairy industries. The study aimed to evaluate microbiological parameters and detect multi-antibiotic-resistant pathogens at three specific supply points and to explore the association between the presence of residual antibiotics and the resistant isolates in milk samples. About 50 milk samples, including raw, soon-after-processed, packaged marketed pasteurized, and UHT milk, were subjected to microbiological analysis. This involved assessing the total bacterial count (TBC) and total coliform count (TCC), conducting antibiotic susceptibility tests through disk and well diffusion assays, detecting virulence genes in multi-antibiotic resistant isolates using gene-specific PCR, and analyzing residual antibiotics by HPLC. The study revealed that the quality of raw milk samples was unacceptable (TBC >4.5x107 CFU/mL and TCC >5.6x104 CFU/mL), while pasteurized samples from processing plants had lower counts than those from retail stores (TBC >5x105 CFU/mL and TCC >1.6x104 CFU/mL) indicating post-pasteurization contamination. About 70.37% of the isolates were Gram-negative, with Escherichia coli (21.4%) and Vibrio (18.8%) being the most prevalent. Resistance to antibiotics was substantial, particularly against ampicillin (86.3%), tetracycline (76%), and ciprofloxacin (58.9%). Gene-specific PCR analysis detected uidA, oprL, and oprI virulence genes in multi-drug-resistant Escherichia coli and Pseudomonas sp. respectively. The study also revealed a direct association between the presence of residual antibiotics and the resistant isolates, emphasizing the need for dairy industry improvements. As high bacterial counts in milk can pose health risks by fostering antibiotic-resistant pathogens, it is essential to mitigate microbiological contamination in the milk supply chain through the implementation of various precautionary measures.

References

Adzitey, F. et al. (2022) ‘Antimicrobial Susceptibility and Molecular Characterization of Escherichia coli Recovered from Milk and Related Samples’, Microorganisms, 10(7). Available at:

https://doi.org/10.3390/MICROORGANISMS10071335.

Akter, T. et al. (2021) ‘Raw Milk In Noakhali, Bangladesh: Quality Assessment and Antibiotic Resistance of Identified Microorganisms’, Current Research in Nutrition and Food Science, 9(3), pp. 1104–1112. Available at: https://doi.org/10.12944/CRNFSJ.9.3.35.

Anika, T.T. et al. (2019) ‘Time dependent screening of antibiotic residues in milk of antibiotics treated cows’, Journal of Advanced Veterinary and Animal Research, 6(4), p. 516.

Arafat, M. et al. (2015) ‘Quality of ultra-high temperature treated milk available in Gazipur and Mymensingh of Bangladesh’, Bangladesh Journal of Animal Science, 44(3), pp. 132–136. Available at:

https://doi.org/10.3329/BJAS.V44I3.26362.

Aslam, M. et al. (2014) ‘Characterization of extraintestinal pathogenic Escherichia coli isolated from retail poultry meats from Alberta, Canada’, International Journal of Food Microbiology, 177, pp. 49–56.

Atia, R.M. et al. (2022) ‘Incidence of pseudomonas specises and effect of their virulence factors on milk and milk products.’, Benha Veterinary Medical Journal, 42(1), pp. 1–5. Available at:

https://doi.org/10.21608/BVMJ.2022.103086.1481.

Banik, S.K., Das, K.K. and Uddin, M.A. (2014) ‘Microbiological quality analysis of raw, pasteurized, UHT milk samples collected from different locations in Bangladesh’, Stamford Journal of Microbiology, 4(1), pp. 5–8. Available at: https://doi.org/10.3329/SJM.V4I1.22753.

BFSA (2021) law - Bangladesh Food Safety Authority-Food Safety (Determination and Control of Pathogenic Microorganisms) Regulations, 2021, bfsa.gov.bd. Dhaka. Available at: http://bfsa.gov.bd/site/view/law (Accessed: 20 March 2023).

Bhaurao, B.C., Rajendra, A.S. and Sarita, L.S. (2022) ‘Isolation and characterization of multidrug resistance bacteria from hospital sewage samples, Maharashtra, India’, African Journal of Biotechnology , 21(1), pp. 16–25. Available at: https://doi.org/10.5897/AJB2021.17394.

Brown, K. et al. (2020) ‘Antibiotic residues and antibiotic-resistant bacteria detected in milk marketed for human consumption in Kibera, Nairobi’, Plos one, 15(5), p. e0233413.

BSTI (2002) Bangladesh Standard: Specification for Pasteurized Milk. Dhaka.

Buczkowska, M. et al. (2021) ‘Penicillin and tetracycline residues in selected fresh and UHT milk with different fat contents’, International Food Research Journal, 28(4), pp. 780–787. Available at: https://doi.org/10.47836/IFRJ.28.4.14.

CLSI (2020) Performance Standards for Antimicrobial Susceptibility Testing .

Dashti, A.A. and Dashti, H. (2009) ‘Heat Treatment of Bacteria: A Simple Method of DNA Extraction for Molecular Techniques’, Article in The Journal of the Kuwait Medical Association [Preprint]. Available at: https://www.researchgate.net/publication/266888615 (Accessed: 29 April 2023).

Dhanashekar, R., Akkinepalli, S. and Nellutla, A. (2012) ‘Milk-borne infections. An analysis of their potential effect on the milk industry’, Germs, 2(3), p. 101. Available at: https://doi.org/10.11599/GERMS.2012.1020.

Economou, V. and Gousia, P. (2015) ‘Agriculture and food animals as a source of antimicrobial-resistant bacteria’, Infection and Drug Resistance, 8, p. 49. Available at: https://doi.org/10.2147/IDR.S55778.

Frazier, W.C. and Westhoff, D.C. (2007) Food Microbiology. Fourth. Edited by T. McGraw-Hill.

Hassani, S. et al. (2022) ‘High prevalence of antibiotic resistance in pathogenic foodborne bacteria isolated from bovine milk’, Scientific Reports, 12(1), p. 3878.

Hossain, T.J., Alam, M.K. and Sikdar, D. (2011) ‘Chemical and microbiological quality assessment of raw and processed liquid market milks of Bangladesh’, Continental journal of food science and technology, 5(2), pp. 6–17.

Howard, F. (2022) Dairy Supply Chain Continues to Face Challenges | Dairy Herd, Dairy Herd Management. Available at: https://www.dairyherd.com/news/business/dairy-supply-chain-continues-face-challenges (Accessed: 7 April 2023).

Hussain, Syed Ahmed Shahzaeem et al. (2023) ‘Antibiotic Misuse in South Asia: A Short Communication Report’, Asia-Pacific Journal of Public Health, 35(1), pp. 82–84. Available at:https://doi.org/10.1177/10105395221149252/ASSET/10105395221149252.FP.PNG_V03.

Islam, M.A. et al. (2018) ‘Microbiological quality assessment of milk at different stages of the dairy value chain in a developing country setting’, International Journal of Food Microbiology, 278, pp. 11–19.

Jamal, J.B., Akter, S. and Uddin, M.A. (2018) ‘Microbiological quality determination of pasteurized, UHT and flavoured milk sold in Dhaka, Bangladesh’, Stamford Journal of Microbiology, 8(1), pp. 1–6.

Jayarao, B.M. and Wang, L. (1999) ‘A Study on the Prevalence of Gram-Negative Bacteria in Bulk Tank Milk’, Journal of Dairy Science, 82(12), pp. 2620–2624. Available at: https://doi.org/10.3168/JDS.S0022-0302(99)75518-9.

Kamaruzzaman, E.A. et al. (2020) ‘Occurrence and characteristics of extended-spectrum β-lactamase-producing Escherichia coli from dairy cattle, milk, and farm environments in Peninsular Malaysia’, Pathogens, 9(12), p. 1007.

Kumar, A., Tolossa, D. and Abdisa, M. (2015) ‘Assessment of Raw Milk Microbial Quality at Different Critical Points of Oromia to Milk Retail Centers in Addis Ababa’, Food Science and Quality Management, 38(0), pp. 1–9. Available at: https://www.iiste.org/Journals/index.php/FSQM/article/view/21355 (Accessed: 9 April 2023).

Miciński, J. et al. (2012) ‘The effects of bovine milk fat on human health’, Polish Annals of Medicine, 19(2), pp. 170–175. Available at: https://doi.org/https://doi.org/10.1016/j.poamed.2012.07.004.

Mokhtari, A. and Amini, K. (2019) ‘Genotyping of Pseudomonas Aeruginosa Strains As A Multidrug Resistant (MDR) Bacterium And Evaluating The Prevalence of Esbls and Some Virulence Factors Encoding Genes By PFGE and ERIC-PCR Methods’, Iranian Journal of Pharmaceutical Research : IJPR, 18(3), p. 1580. Available at: https://doi.org/10.22037/IJPR.2019.1100762.

Momtaz, H. et al. (2013) ‘Detection of Escherichia coli, Salmonella species, and Vibrio cholerae in tap water and bottled drinking water in Isfahan, Iran’, BMC Public Health, 13(1), pp. 1–7. Available at: https://doi.org/10.1186/1471-2458-13-556/TABLES/5.

Naganboyina, T. and Kaple, Prof.G. (2022) ‘A Study On Dairy Supply Chain Management In India – Its Development, Policies & Barriers’, Journal of Positive School Psychology, 6, pp. 1977–1984.

Nikbin, V.S. et al. (2012) ‘Molecular identification and detection of virulence genes among Pseudomonas aeruginosa isolated from different infectious origins’, Iranian journal of microbiology, 4(3), p. 118.

Nirwal, S., Pant, R. and Rai, N. (2013) ‘Analysis of milk quality, adulteration and mastitis in milk samples collected from different regions of Dehradun’, International Journal of PharmTech Research, 5(2), pp. 359–364.

Nowar, A. et al. (2021) ‘Microbiological Quality Assessment and Identification of Antibiotic Resistant Bacteria at Different Stages of the Milk Supply Chain in Dhaka City of Bangladesh’, Journal of Advances in Microbiology, 21(10), pp. 67–76.

Ntuli, V., Njage, P.M.K. and Buys, E.M. (2016) ‘Characterization of Escherichia coli and other Enterobacteriaceae in producer-distributor bulk milk’, Journal of dairy science, 99(12), pp. 9534–9549.

Nur, I.T. et al. (2021) ‘Microbiological Quality Assessment of Milk and Milk Products Along with their Packaging Materials Collected from a Food Industry in the Dhaka DivisionMicrobiological Quality Assessment of Milk and Milk Products Along with their Packaging Materials Collected from a Food Industry in the Dhaka Division’, SVOA Microbiology, (2634–534X), pp. 19–25.

Oliveira, J. and Reygaert, W.C. (2022) ‘Gram Negative Bacteria’, Infection Management for Geriatrics in Long-Term Care Facilities, Second Edition, pp. 427–443. Available at: https://doi.org/10.1385/1-59259-036-5:43.

Peters, L. et al. (2019) ‘Multiple antibiotic resistance as a risk factor for mortality and prolonged hospital stay: a cohort study among neonatal intensive care patients with hospital-acquired infections caused by gram-negative bacteria in Vietnam’, PloS one, 14(5), p. e0215666.

Rahman, M.S., Hassan, M.M. and Chowdhury, S. (2021) ‘Determination of antibiotic residues in milk and assessment of human health risk in Bangladesh’, Heliyon, 7(8). Available at: https://doi.org/10.1016/J.HELIYON.2021.E07739.

Rice, E.W., Bridgewater, L. and Association, A.P.H. (2012) Standard methods for the examination of water and wastewater. American public health association Washington, DC.

Rokon-Uz-Zaman, Md. et al. (2023) ‘Detection of antimicrobial resistance genes in Lactobacillus spp. from poultry probiotic products and their horizontal transfer among Escherichia coli’, Veterinary and Animal Science, 20, p. 100292. Available at: https://doi.org/10.1016/J.VAS.2023.100292.

Sainz-Mejías, M., Jurado-Martín, I. and McClean, S. (2020) ‘Understanding Pseudomonas aeruginosa–host interactions: The ongoing quest for an efficacious vaccine’, Cells, 9(12), p. 2617.

Sarowska, J. et al. (2019) ‘Virulence factors, prevalence and potential transmission of extraintestinal pathogenic Escherichia coli isolated from different sources: recent reports’, Gut pathogens, 11, pp. 1–16.

Sebastião, F.A. et al. (2023) ‘Antimicrobial resistance profile of Aeromonas spp. isolated from asymptomatic Colossoma macropomum cultured in the Amazonas State, Brazil’, Brazilian Journal of Biology, 82, p. e260773. Available at: https://doi.org/10.1590/1519-6984.260773.

Siljanoski, A. et al. (2018) ‘Detection of tetracycline and other antimicrobial residues in milk from cows with clinical mastitis treated by combination therapy’, Journal of Dairy Research, 85(3), pp. 321–326. Available at: https://doi.org/10.1017/S0022029918000389.

Skalet, A.H. et al. (2010) ‘Antibiotic Selection Pressure and Macrolide Resistance in Nasopharyngeal Streptococcus pneumoniae: A Cluster-Randomized Clinical Trial’, PLOS Medicine, 7(12), p. e1000377. Available at: https://doi.org/10.1371/JOURNAL.PMED.1000377.

Smith, J.L., Fratamico, P.M. and Gunther, N.W. (2007) ‘Extraintestinal pathogenic Escherichia coli’, Foodborne pathogens and disease, 4(2), pp. 134–163.

Sobeih, A.M.K. et al. (2020) ‘Prevalence oF Enterobacteriaceae in raw milk and some dairy products’, Kafrelsheikh Veterinary Medical Journal, 18(2), pp. 9–13. Available at: https://doi.org/10.21608/KVMJ.2020.39992.1009.

Tamirat, T. (2018) ‘Microbiological quality analysis of raw and pasteurized milk samples collected from Addis Ababa and its surrounding in Ethiopia’, Appro Poult Dairy and Vet Sci, 4, pp. 0–8.

Tekinsen, K.K., Elmali, M. and Ulukanli, Z. (2007) ‘Microbiological quality of UHT milk consumed in Turkey’, Internet J. Food Safety, 7, pp. 45–48.

Ventola, C.L. (2015) ‘The Antibiotic Resistance Crisis: Part 1: Causes and Threats’, Pharmacy and Therapeutics, 40(4), p. 277. Available at: https://doi.org/Article.

Wall, B.A. et al. (2016) ‘Drivers, dynamics and epidemiology of antimicrobial resistance in animal production’, p. 58. Available at: https://doi.org/10.3/JQUERY-UI.JS.

Zahreddine, Z. et al. (2021) ‘HPLC-DAD multi-residue method for determination of florfenicol, penicillin and tetracycline residues in raw cow milk’, Journal of Clinical and Laboratory Research, 2(3), pp. 2487–2768.

Published

31-12-2023

How to Cite

Rinky, F., Reza, S., Nowar, A., Ghosh, S., Rahman, A., & Alim, S. R. (2023). ANALYSIS OF MICROBIOLOGICAL QUALITY AND ANTIBIOTIC RESISTANCE PATTERNS IN MILK SUPPLY CHAIN: MICROBIOLOGICAL QUALITY AND ANTIBIOTIC RESISTANCE PATTERNS IN MILK. Bioresearch Communications - (BRC), 10(01), 1462–1473. https://doi.org/10.3329/brc.v10i1.70686

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Original Article