Veterinary Research Forum

Genetic characterization of virulence and extended spectrum β-lactamase producing genes of Klebsiella pneumoniae isolated from bovine milk

Document Type : Original Article

Authors

Bhavinkumar Pankajbhai Katira 1
Bhaveshkumar Ishwarbhai Prajapati 1
Ratn Deep Singh 2
Sandipkumar Sureshbhai Patel 3
Kirankumar Motiji Solanki 1

1 Department of Veterinary Public Health and Epidemiology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Sardarkrushinagar, India

2 Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Sardarkrushinagar, India

3 Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Sardarkrushinagar, India

https://doi.org/10.30466/vrf.2023.2006974.3947
Abstract
Mastitis associated Klebsiella pneumoniae species were isolated from bovine milk to characterize virulence genes (wabG and kfuBC) and extended spectrum β-lactamase (ESBL) genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaTEM, blaSHV and blaOXA). A total number of 325 bovine milk samples (195 raw and 130 mastitic milk specimens) collected from Banaskantha, a milk-shed district of Gujarat, India, were included in the study. A total number of 27 K. pneumoniae isolates were recovered, consisting of 17 (62.96%) isolates from raw milk and 10 (37.03%) isolates from mastitic milk samples, giving an overall prevalence of 8.31%. Antibiotic sensitivity patterns revealed that 20 out of 27 isolates were found to be multi-drug resistant. Based on combination disc diffusion test and HiCrome ESBL agar method, 20 (74.07%) and 25 (92.59%) isolates were detected as ESBL producers, respectively. Among virulence genes studied, presence of wabG (25/27; 92.59%) was higher than kfuBC (5/27; 18.51%). Beta-lactamase genes viz., blaSHV, blaTEM and blaCTX-M-1 were detected in 23/27 (85.18%), 3/27 (11.11%) and 2/27 (7.40%) of isolates, respectively; while, none of the isolates was found to be positive for blaCTX-M-9 and blaOXA-1 genes. Outcome of the study provided an insight into virulence genes and ESBL producing K. pneumoniae isolated from bovine milk samples in India.

Keywords

Bovine mastitis
Drug resistance
India
Klebsiella pneumoniae
Virulence factors
Subjects
  1. Milk production in India. Available at: https://pib.gov. in/FeaturesDeatils.aspx?NoteId=151137. Accessed July 18, 2023.
  2. Massé J, Dufour S, Archambault M. Characterization of Klebsiella isolates obtained from clinical mastitis cases in dairy cattle. J Dairy Sci 2020; 103(4): 3392-3400.
  3. Huang S, Ji S, Wang F, et al. Dynamic changes of the fecal bacterial community in dairy cows during early lactation. AMB Express 2020; 10(1): 167. doi: 10.1186/ s13568-020-01106-3.
  4. Langoni H, Guiduce MVS, Nóbrega DB, et al. Research of Klebsiella pneumoniae in dairy herds. Pesqui Vet Bras 2015; 35(1): 9-12.
  5. Sugiyama M, Watanabe M, Sonobe T, et al. Efficacy of antimicrobial therapy for bovine acute Klebsiella pneu-moniae J Vet Med Sci 2022; 84(7): 1023-1028.
  6. Hasani A, Soltani E, Rezaee MA, et al. Serotyping of Klebsiella pneumoniae and its relation with capsule-associated virulence genes, antimicrobial resistance pattern, and clinical infections: a descriptive study in medical practice. Infect Drug Resist 2020; 13: 1971-1980.
  7. Elasmer AH, Ibrahim MY, Rizk DE. Seroepidemiology, antimicrobial susceptibility and virulence characteristics of clinical Klebsiella pneumoniae isolates in Mansoura university hospitals. Egypt J Med Microbiol 2021; 30(1): 9-18.
  8. Zheng Z, Gorden PJ, Xia X, et al. Whole-genome analysis of Klebsiella pneumoniae from bovine mastitis milk in the US. Environ Microbiol 2022; 24(3): 1183-1199.
  9. Prajapati BI, Solanki KM, Devi S, et al. Phenotypic and molecular characterization of ESBLs producing Escherichia coli in bovine faecal and milk samples of North Gujarat. Indian J Anim Sci 2020; 90(7): 987-992.
  10. Riwu KHP, Effendi MH, Rantam A review of extended spectrum β-lactamase producing Klebsiella pneumoniae and multidrug resistant (MDR) on companion animals. Syst Rev Pharm 2020; 11(7): 270-277.
  11. Theocharidi NA, Balta I, Houhoula D, et al. High prevalence of Klebsiella pneumoniae in Greek meat products: detection of virulence and antimicrobial resistance genes by molecular techniques. Foods 2022; 11(5): 708. doi: 10.3390/foods11050708.
  12. Yang F, Zhang S-D, Shang X-F, et al. Prevalence and characteristics of extended spectrum β-lactamase-producing Escherichia coli from bovine mastitis cases in China. J Integr Agric 2018; 17(6): 1246-1251.
  13. Koovapra S, Bandyopadhyay S, Das G, et al. Molecular signature of extended spectrum β-lactamase producing Klebsiella pneumoniae isolated from bovine milk in eastern and north-eastern India. Infect Genet Evol 2016; 44: 395-402.
  14. Bandyopadhyay S, Banerjee J, Bhattacharyya D, et al. Genomic identity of fluoroquinolone-resistant bla CTX-M-15-Type ESBL and pMAmpC β-lactamase producing Klebsiella pneumoniae from buffalo milk, India. Microb Drug Resist 2018; 24(9): 1345-
  15. Quinn PJ, Markey BK, Leonard FC, et al. Veterinary microbiology and microbial disease. 2nd Chichester, UK: John Wiley & Sons 2011; 263-286.
  16. Younis G, Sadat A, Maghawry M. Characterization of coa gene and antimicrobial profiles of Staphylococcus aureus isolated from bovine clinical and subclinical mastitis. Adv Anim Vet Sci 2018; 6(4): 161-168.
  17. Performance standards for antimicrobial susceptibility testing. 28th informational supplement. M100-S28. Wayne, Pennsylvania: Clinical and Laboratory Standards Institute, 2018.
  18. Brisse S, Verhoef J. Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping. Int J Syst Evol Microbiol 2001; 51(Pt 3): 915-924.
  19. Chander Y, Ramakrishnan MA, Jindal N, et al. Differentiation of Klebsiella pneumoniae and oxytoca by multiplex polymerase chain reaction. Int J Appl Res Vet Med 2011; 9(2): 138-142.
  20. Brisse S, Fevre C, Passet V, et al. Virulent clones of Klebsiella pneumoniae: Identification and evolutionary scenario based on genomic and phenotypic characterization. PloS One 2009; 4(3): e4982. doi: 10.1371/journal.pone.0004982.
  21. Dallenne C, Da Costa A, Decré D, et al. Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteri-aceae. J Antimicrob Chemother 2010; 65(3): 490-4
  22. Kar D, Bandyopadhyay S, Bhattacharyya D, et al. Molecular and phylogenetic characterization of multi-drug resistant extended spectrum beta-lactamase producing Escherichia coli isolated from poultry and cattle in Odisha, India. Infect Genet Evol 2015; 29: 82-90.
  23. El-Sukhon SN. Identification and characterization of Klebsiellae isolated from milk and milk products in Jordan. Food Microbiol 2003; 20(2): 225-230.
  24. Martin RM, Bachman MA. Colonization, infection, and the accessory genome of Klebsiella pneumoniae. Front Cell Infect Microbiol 2018; 8: 4. doi: 10.3389/fcimb. 2018.00004.
  25. Osman KM, Hassan HM, Orabi A, et al. Phenotypic, antimicrobial susceptibility profile and virulence factors of Klebsiella pneumoniae isolated from buffalo and cow mastitic milk. Pathog Glob Health 2014; 108(4): 191-199.
  26. Enferad E, Mahdavi Antibiotic resistance pattern and frequency of some beta lactamase genes in Klebsiella pneumoniae isolated from raw milk samples in Iran. J Hellenic Vet Med Soc 2020; 71(4): 2455-2462.
  27. Yang Y, Peng Y, Jiang J, et al. Isolation and characterization of multidrug-resistant Klebsiella pneumoniae from raw cow milk in Jiangsu and Shandong provinces, China. Transbound Emerg Dis 2021; 68(3): 1033-1039.
  28. Wu X, Liu J, Feng J, et al. Epidemiology, environmental risks, virulence, and resistance determinants of Klebsiella pneumoniae from dairy cows in Hubei, China. Front Microbiol 2022; 13: 858799. doi: 10.3389/ fmicb.2022.858799.
  29. Badri AM, Ibrahim IT, Mohamed SG, et al. Prevalence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae isolated from raw milk samples in Al Jazirah state, Sudan. Mol Biol 2017; 7(1): 201. doi:10.4172/2168-9547.1000201.
  30. Wareth G, Neubauer H. The Animal-foods-environment interface of Klebsiella pneumoniae in Germany: an observational study on pathogenicity, resistance development and the current situation. Vet Res 2021; 52: 16. doi: 10.1186/s13567-020-00875-w.
  31. Bobbadi S, Chinnam BK, Nelapati S, et al. Occurrence and genetic diversity of ESBL producing Klebsiella species isolated from livestock and livestock products. J Food Saf 2020; 40(1): e12738. doi: 10.1111/jfs.12738.
  32. Younis AI, Elbialy AI, Abo Remila EM, et al. Molecular detection of genus Klebsiella and genotypic identification of Klebsiella pneumoniae and Klebsiella oxytoca by duplex polymerase chain reaction in poultry. Glob Vet 2017; 18(3): 234-241.
  33. Wyres KL, Lam MMC, Holt KE. Population genomics of Klebsiella pneumoniae. Nat Rev Microbiol 2020; 18: 344-359.
  34. He Y, Guo X, Xiang S, et al. Comparative analyses of phenotypic methods and 16S rRNA, khe, rpoB genes sequencing for identification of clinical isolates of Klebsiella pneumoniae. Antonie Van Leeuwenhoek 2016; 109(7): 1029-1040.
  35. Razmyar J, Zamani AH. An outbreak of yolk sac infection and dead-in-shell mortality in common canary (Serinuscanaria) caused by Klebsiella pneumoniae. Iran J Vet Res 2016; 17(2): 141-143.
  36. Memon J, Kashif J, Yaqoob M, et al. Molecular characterization and antimicrobial sensitivity of pathogens from sub-clinical and clinical mastitis in Eastern China. Pak Vet J 2013; 33(2): 170-174.
  37. Chehabi CN, Nonnemann B, Astrup LB, et al. In vitro antimicrobial resistance of causative agents to clinical mastitis in Danish dairy cows. Foodborne Pathog Dis 2019; 16(8): 562-572.
  38. Cheng F, Li Z, Lan S, et al. Characterization of Klebsiella pneumoniae associated with cattle infections in southwest China using multi-locus sequence typing (MLST), antibiotic resistance and virulence-associated gene profile analysis. Braz J Microbiol 2018; 49(Suppl 1): 93-100.
  39. Zhang S, Yang G, Ye Q, et al. Phenotypic and genotypic characterization of Klebsiella pneumoniae isolated from retail foods in China. Front Microbiol 2018; 9: 289. doi: 10.3389/fmicb.2018.00289.
  40. Remya PA, Shanthi M, Sekar U. Characterisation of virulence genes associated with pathogenicity in Klebsiella pneumoniae. Indian J Med Microbiol 2019; 37(2): 210-218.
  41. Cheng J, Zhou M, Nobrega DB, et al. Genetic diversity and molecular epidemiology of outbreaks of Klebsiella pneumoniae mastitis on two large Chinese dairy farms. J Dairy Sci 2021; 104(1): 762-775.
  42. Locatelli C, Scaccabarozzi L, Pisoni G, et al. CTX-M1 ESBL-producing Klebsiella pneumoniae pneumoniae isolated from cases of bovine mastitis. J Clin Microbiol 2010; 48(10): 3822-3823.
  43. Sudarwanto M, Akineden Ö, Odenthal S, et al. Extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae in bulk tank milk from dairy farms in Indonesia. Foodborne Pathog Dis 2015; 12(7): 585-590.
Veterinary Research Forum
Volume 15, Issue 2
February 2024
Pages 57-64

  • Receive Date 20 July 2023
  • Revise Date 29 August 2023
  • Accept Date 23 September 2023

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