3-Title: Phenotypic and genotypic characterization of extended spectrum beta-lactamases producing Klebsiella pneumoniae in bovines from Jammu region, India

Authors: Deep Shikha, Virendera Singh Wazir, Mohd Rashid and Sabahat Gazal

Source: Ruminant Science (2021)-10(2):253-259.

How to cite this manuscript: 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.

Abstract

The study was conducted to determine the phenotypic and genotypic characterization of Extended Spectrum Beta-Lactamases producing Klebsiella in bovines from Jammu region, India. Out of the total 30 ESBL isolates, resistance to cefotaxime and ceftazidime was observed in 60.0% (15) and 40.0% (10) isolates, respectively while 64.0% (16) isolates showed resistance to both. The present study revealed that the total prevalence of ESBL producing Klebsiella pneumonia in bovines of the Jammu region is 83.33% and is reported for the first time in Jammu region. Out of ESBL isolates, only 44.0% (11) isolates carried the gene/s for blaTEM, blaCTX-M and blaSHV genes by m-PCR. Out of 11 isolates, only 18.18% (2) isolates carried blaTEM gene alone, 54.54% (6) isolates carried blaCTX-M gene alone, 18.18% (2) isolates carried both blaTEM/blaCTX-M genes and only 9.09% (1) isolates carried blaSHV /blaTEM /blaCTX-M. Of the six blaCTX-M positive isolates, 50.0% (3) belonged to CTX-M-1 group, 33.33% (2) to CTX-M-9 group and 16.66% (1) isolates carried both blaCTX-M 1 / blaCTX-M 9. One representative amplicon of blaSHV, blaTEM and blaCTX-M were cloned in pGEM-T and pJET1.2 and got sequenced. The sequence of blaCTX-M was found to match with the database sequence of blaCTX-M -15 and is the first report of its kind in India. Antimicrobial sensitivity test of 11 ESBL producing Klebsiella isolates showed that resistance was most commonly to ampicillin (100%), cefexime (100%), neomycin (100%) followed by streptomycin (90.90%). Antibiotic chloramphenicol (81.81%) and imipenem (72.72%) show sensitive results. Out of 11 multi-drug resistance ESBL positive isolates, only 07 isolates tested positive for class 1 and 2 integrons. Among the 07, 57.14% (4) isolates harbour intI 1 gene, while 28.57% (2) isolates carried intI 2 gene. One (14.28%) isolate, was found to harbour both intI 1 and intI 2.

References

Carole GMV, Kouadio GN, Baguy OM, Djeneba OG, Ayayi A, Bertin TK, Anatole TA, Innocent KK, Kpoda DS, Eric T, Ali K, Fernique K, Alphonse K, Mireille D and Bmr G (2018). Antimicrobial resistance profile and molecular characterization of extended-spectrum beta-lactamase genes in enterobacteria isolated from human, animal and environment. Journal of Advances in Microbiology 10(1):1-9.

Chen T, Feng Y, Yuan JL, Qi Y, Cao YX and Wu Y (2013). Class 1 integrons contributes to antibiotic resistance among clinical isolates of Escherichia coli producing extended-spectrum beta-lactamases. Indian Journal of Medical Microbiology 31(4):385-389.

Collis CM, Kim MJ, Stokes HW and Hall RM (2002). Integron encoded IntI integrases preferentially recognize the adjacent cognate attI site in recombination with a 59-be site. Molecular Microbiology 46:1415-1427.

Correa FE, Dantas FG, Grisolia AB, CrispimBdo A and Oliveira KM (2014). Identiûcation of class 1 and 2 integrons from clinical and environmental Salmonella isolates. Journal of Infection in Developing Countries 8:1518-1524.

Das L, Borah P, Sharma RK, Malakar D, Saikia GK, SharmaK, Tamuly S and Dutta R (2020). Phenotypic and molecular characterization of extended spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae isolates from various samples of animal origin from Assam, India. bioRxiv.

Ejaz H, Younas S, Abosalif KOA, Junaid K, Alzahrani B and Alsrhani A (2021). Molecular analysis of blaSHV, blaTEM, and blaCTX-M in extended-spectrum beta-lactamase producing Enterobacteriaceae recovered from fecal specimens of animals. Public Library of Science ONE 16(1):1-15.

Fang H, Ataker F, Hedin G and Dornbusch K (2008). Molecular epidemiology of extended-spectrum beta-lactamases among Escherichia coli isolates collected in a Swedish hospital and its associated health care facilities from 2001 to 2006. Journal of Clinical Microbiology 46 (2):707-712.

Fateh S, Hirpurkar SD, Shakya S, Rawat N, Devangan P, Khan FF and Bhandekar K (2017). Presence of enterobacteria producing extended-spectrum beta-lactamases and/or carbapenemases in animals, humans and environment in India. Thai Journal of Veterinary Medicine 47(1):35-43.

Haennia M, Chatre P, Metayera V, Bour M, Signol E, Madec JY and Gay E (2014).  Comparative prevalence and characterization of ESBL-producing Enterobacteriaceae in dominant versus subdominant enteric flora in veal calves at slaughterhouse, France. Veterinary Microbiology 6527:7.

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.

Jain Shweta, Rajput Neetu and Sahni YP (2021). Disposition kinetics and dosage regimen of cefepime following single intravenous administration in healthy and febrile buffalo calves. Ruminant Science 10(1):143-146.

Kpoda DS, Ajayi A, Somda M, Traore O, Guessennd N, Ouattara AS, Sangare L, Traore A.S and Dosso M  (2018). Distribution of resistance genes encoding ESBLs in Enterobacteriaceae isolated from biological samples in health centers in Ouagadougou, Burkina Faso. BMC Research Notes 11:471.

Machado E., Canton, R, Baquero F, Galan JC, Rollan A, Peixe L and Coque TM (2005). Integron content of extended-spectrum-beta-lactamase-producing Escherichia coli strains over 12 years in a single hospital in Madrid, Spain. Antimicrobial Agents and Chemotherapy 49(5):1823-1829.

Mazel  D ( 2006). Integrons: Agents of bacterial evolution. Nature Reviews Microbiology 4:608- 620.

Montso P, Dlamini SB, Kumar A and Ateba CN (2019). Antimicrobial resistance factors of extended-spectrum beta-lactamases producing Escherichia coli and Klebsiella pneumoniae isolated from cattle farms and raw beef in north-west province, South Africa Kotsoana. BioMedical Research 13.

Olowe OA,  Adewumi O, Odewale G, Ojurongbe O and Adefioye OJ (2015). Phenotypic and molecular characterization of extended-spectrum beta-lactamase producing Escherichia coli obtained from animal fecal samples in Ado Ekiti, Nigeria. Journal of Environmental and Public Health 1-7.

Paterson DP and Bonomo RA (2005). Extended-spectrum beta-lactamases: A Clinical Update. Clinical Microbiology Reviews 657-686.

Qamsari MM, Panah MA, Eftekhar F and Feizabadi MM (2013). Integron mediated multidrug resistance in extended spectrum beta-lactamase producing clinical isolates of Klebsiella pneumoniae. Brazilian Journal of Microbiology 44(3):849-854.

Salem MM, Muharram M and Alhosiny IM (2010). Distribution of classes 1 and 2 integrons among multi-drug resistant E. coli isolated from hospitalized patients with urinary tract infection in Cairo, Egypt. Australian Journal of Basic and Applied Sciences 43:398-407.

Shabaz SS, Prameela DR, Sreenivasulu D and Sujatha K (2020). Evaluation of on-farm milk culture system for identification of mastitis pathogens. Ruminant Science 9(2):363-372.

Stokes HW and Hall RM (1989). A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: Integrons. Molecular Microbiology 3:1669-1683.

Subramanya SH, Bairy I, Metok Y, Baral BP, Gautam D and  Nayak N (2021). Detection and characterization of ESBL producing Enterobacteriaceae from the gut of subsistence farmers, their livestock, and the surrounding environment in rural Nepal. Scientific Reports 11:2091.

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.

Umadevi S, Kandhakumari G, Joseph NM, Kumar S, Easow JM, Stephen S and Singh UK (2011). Prevalence and antimicrobial susceptibility pattern of ESBL producing Gram negative bacilli. Journal of Clinical and Diagnostic Research 5(2):236-239.

Woodford N and Sundsfjord A (2005). Molecular detection of antibiotic resistance: When and where. Journal of Antimicrobial Chemotherapy 56:259-261.

Yan H, Li L, Zong M, Alam MJ, Shinoda S and Shi L (2010). Occurrence and characteristics of class 1 and 2 integrons in clinical bacterial isolates from patients in South China. Journal of Health Sciences 56:442-450.