20-Title: Detection of extended-spectrum beta-lactamase and carbapenemase resistant Klebsiella pneumoniae in bovine milk
Authors: Sangita Choudhary, Bincy Joseph, Sandeep Kumar Sharma, Surendra Singh Shekhawat and Nazeer Mohammed
Source: Ruminant Science (2022)-11(1):93-100.
How to cite this manuscript: Choudhary Sangita, Joseph Bincy, Sharma Sandeep Kumar, Shekhawat Surendra Singh and Mohammed Nazeer (2022). Detection of extended-spectrum beta-lactamase and carbapenemase resistant Klebsiella pneumoniae in bovine milk. Ruminant Science 11(1):93-100.
The present study aimed at the screening of ESBL resistance in the Klebsiella pneumoniae organism isolated from bovine milk. A total of 109 milk samples were collected randomly from the urban and peri-urban areas of two major cities of Rajasthan, Jodhpur and Jaipur. The study included milk samples from healthy as well as from animals with clinical mastitis. From all the samples 41 isolates of Klebsiella pnuemoniae were obtained which were characterized morphologically, biochemically and using molecular methods. All 41 isolates were subjected to an antibiotic sensitivity test and phenotypic and genotypic ESBL resistance test. Most of the isolates were resistant to more than 6 antibiotics used. The isolate number RJOBK98 and RJOCK10 were resistant to 21 antibiotic used and ten isolates (RJABK12, RJABK16, RJABK21, RJACK22, RJACK53, RJOCK66, RJOBK67, RJABK75, RJOCK94, RJABK107) were resistant to 20 antibiotics used. All the isolates (41) showed (100%) resistance to Penicillin- G (P10), Ceftazidine / Clavulanate (CAC30/10), Vancomycin (VA30). Out of the 41 isolates, 40 (97.56%) were resistant to Cefazolin (CZ30) and Cefotaxime (CTX30). Here 100% of isolates showed a MAR index of more than 0.2% which indicates that all the isolates had a high potential of developing multiple antibiotic resistance and the ability to transfer the resistance to others. 35 isolates were positive for phenotypic ESBL production by double disc diffusion test irrespective of the city, species, or the clinical condition of the animal. Among the three ESBL genes (blaTEM, blaSHV and blaCTX-M) screened blaTEM was present in all the 35 ESBL positive isolates. Two isolates harboured more than 1 ESBL resistant gene (blaSHV or blaCTX-M) in addition to blaTEM. The isolate RJABK 21 carried the blaSHV gene and RJOCK94 carries the blaCTX-M gene in addition to the blaTEM gene. None of the isolates carried all three genes. The isolate number RJABK 21 from Jaipur carried the gene for carbapenem resistance blaKPC and blaNDM. The presence of an emerging carbapenemase resistance gene blaNDM and blaKPC in milk samples is challenging to public health as there is an increased possibility of transmission to human beings through the food chain.
Bandyopadhyay S, Banerjee J, Bhattacharyya D, Samanta I, Mahanti A, Dutta TK, Ghosh S, Nanda PK, Dandapat P and Bandyopadhyay S (2018). Genomic identity of fluoroquinolone-resistant bla CTX-M-15-Type ESBL and pMAmpC beta-lactamase producing Klebsiella pneumoniae from buffalo milk India. Microbial Drug Resistance 24(9):1345-1353.
Bhati Taruna, Chhabra Rajesh, Yadav Rahul, Charaya Gaurav and Kataria Anil Kumar (2021). Antimicrobial resistance profiling of Staphylococcus aureus isolated from mastitic milk of bovine and dairy environment from arid and semi-arid regions of India. Ruminant Science 10(1):57-66.
Bonardi S and Pitino R (2019). Carbapenemase-producing bacteria in food-producinganimals, wildlife and environment: A challenge for human health. Italian Journal of Food Safety 8(2).
CLSI (2017). Performance Standards for Antimicrobial Susceptibility Testing. 27th Edn, Clinical and Laboratory Standard Institute (CLSI) S, M100 Wayne, PA
Dahmen S, Métayer V, Gay E, Madec JY and Haenni M (2013). Characterization of extended-spectrum beta-lactamase (ESBL)-carrying plasmids and clones of Enterobacteriaceae causing cattle mastitis in France. Veterinary Microbiology 162(2-4):793-799.
Ghatak S, Singha A, Sen A, Guha C, Ahuja A, Bhattacharjee U, Das S, Pradhan NR, Puro K, Jana C, Dey TK, Prashant Kumar KL, Das A, Shakuntala I, Biswas U and Jana PS (2013). Detection of New Delhi metallo beta-lactamase and extended-spectrum beta-lactamase genes in Escherichia coli isolated from mastitic milk samples. Transboundary and Emerging Diseases 60(5):385-389.
Jaibhaye CS, Bhikane AU, Masare PS and Bhonsle AV (2020). Clinico-diagnostic and therapeutic investigations on pneumonia in cattle. Ruminant Science 9(1):25-32.
Koovapra S (2015). Characterization of extended-spectrum beta-lactamase-producing Klebsiella spp of bovine mastitis origin. MVSc thesis submitted to IVRI, Izatnagar, UP, India.
Krumperman PH (1983). Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Applied and Environmental Microbiology 46(1):165-170.
Liu Y, Liu C, Zheng W, Zhang X, Yu J, Gao Q, Hao Y and Huang X (2008). PCR detection of Klebsiella pneumoniae in infant formula based on 16S–23Sinternal transcribed spacer. International Journal of Food Microbiology 125(3):230-235.
Nóbrega DB, Guiduce MV, Guimarães FF, Riboli DF, Cunha ML, Langoni H, Pantoja JC and Lucheis SB (2013). Molecular epidemiology and extended-spectrum beta-lactamases production of Klebsiella pneumoniae isolated from three dairy herds. Pesquisa Veterinária Brasileira 33:855-859.
Quinn PJ, Carter ME, Markey BK and Carter GR (1994). General procedures in microbiology. Clinical Veterinary Microbiology Wolfe Publishing, London, p 664.
Shikha Deep, Wazir Virendera Singh, Rashid Mohd and Gazal Sabahat (2021). Phenotypic and genotypic characterization of extended spectrum beta-lactamases producing Klebsiella pneumoniae in bovines from Jammu region, India. Ruminant Science 10(2):253-259.
Sudarwanto M, Akineden Ö, Odenthal S, Gross M and Usleber E (2015). Extended-spectrumâ-lactamase (ESBL)-producing Klebsiella pneumonia in bulk tank milk from dairy farms in Indonesia. Foodborne Pathogen and Disease 12(7):585-590.
Tiwari RK, Galav V, Sharma SK, Galav A, Kumar B, Khan S, Marodia S and Agrawal M (2019). Aetio-pathology of bronchopneumonia in sheep (Ovis aries). Ruminant Science 8(2):217-220.
Wilson K (1987). Preparation of genomic DNA from bacteria. Current Protocol in Molecular Biology. Unit 241, New York, Wiley.
Yadav SK (2020). Phenotypic and genotypic studies on antibiotic resistance pattern of Klebsiella pneumoniae associated with various clinical conditions of animals. MVSc thesis submitted to RAJUVAS, Bikaner, Rajasthan.