The emergence and spread of carbapenem-resistant
Enterobacteriaceae
(CRE) are perceived as a serious public-health threat world-wide. Despite sporadic reports, no systemic study has been carried out ...on CRE in companion animals in Indian subcontinent. In total, 237 canine specimens collected from five veterinary polyclinics in and around Kolkata were analyzed for isolation, antimicrobial resistance profiling and molecular characterization of carbapenem-resistant (CR)
E. coli
. Of the 29 CR isolates, 19 were identified as metallo-β-lactamase producers (MP-CRE) and 10 as metallo-β-lactamase non-producers (MNP-CRE). Eleven of them were extended spectrum β-lactamase and/or AmpC type β-lactamase producers and harboured fluoroquinolone-, tetracycline-, sulfonamide- and aminoglycoside-resistant genes. Beside uropathogenic virulence determinants, they carried the adhesion factors mediating biofilm production which was remarkably higher in 6 MP-CRE and one MNP-CRE isolates. Although the CRE were of diverse origin including the healthy and the diseased dogs, these were more frequently isolated from canine pyometra. The MP-CRE harboured plasmids of IncF and Inc
A/C
types. Phylo-type B1 was observed in 38% of the CR isolates, followed by A0 in 31% and rest were attributed to A1 and D1. The Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) revealed that these isolates were genetically diverse and constituted of a heterogenous population. Detection of CRE in pet dogs despite the fact that carbapenems are not used in animals in India emphasizes the need for active surveillance to identify the transmission and dynamics of such pathogens in companion animals.
A cross-sectional study covering four agro-climatic zones of West Bengal, India, was carried out to understand the risk-factors, antimicrobial resistance mechanism and clustering of the resistance ...characteristics of Escherichia coli isolated from healthy (170) and diarrhoeic (74) goats reared under intensive (52) and semi-intensive (192) farming practices. Of the 488 E. coli isolates, the majority, including the extended spectrum (n: 64, 13.11%) and AmpC β-lactamase (ACBL) (n: 86, 17.62%) producers, were resistant to tetracycline (25.2%), followed by enrofloxacin (24.5%), cefotaxime (21.5%) and amikacin (20.5%). Statistical modelling revealed that the isolates from diarrhoeic animals (p < 0.001) are likely to be more ACBL-positive than those from the healthy counterparts. Similarly, cefotaxime (p < 0.05) and enrofloxacin-resistance (p < 0.01) were significantly higher in diarrhoeic goats and in goats reared intensively. The isolates (n = 35) resistant to multiple drugs revealed the presence of β-lactamase blaCTXM-1-(21), blaSHV-(7), blaTEM-(3), blaCMY-6-(1), blaCITM-(3); quinolone qnrB-(10), qnrS-(7), aac(6’)-Ib-cr-(3); tetracycline tetA-(19), tetB-(4) and sulphonamide resistance determinants sul1-(4); multiple plasmids, especially those belonging to the IncF and IncI1 replicon types; and active acrAB efflux pumps. Further, two isolates harbored the carbapenem resistance (blaNDM-5) gene and eight were strong biofilm producers. This first ever study conducted to unravel the status of AMR in goat farming reveals that not only the intensive farming practices but also certain clinical ailments such as diarrhoea can increase the shedding of the drug-resistant isolate. The emergence of multi-drug resistant (MDR) E. coli in goats, particularly those that are carbapenem resistant, is a cause for concern that indicates the spread of such pathogens even in the livestock sub-sector generally considered as naive.
The present study was conducted to detect the occurrence of methicillin-resistant and enterotoxin producing Staphylococcus aureus in bovine milk with SCCmec, spa type, antimicrobial sensitivity and ...MIC of anti-staphylococcal drugs against the isolates. A total of 450 composite milk samples both from cattle with or without mastitis were collected from all agro-climatic zones of West Bengal. Nine (9.6%) S. aureus isolates were phenotypically detected as MRSA and all of them possessed mecA. Whereas, 12 (12.8%) and 4 (4.3%) isolates possessed the staphylococcal enterotoxins sei and seg, respectively. Six and three isolates belonged to SCCmec type Iva and type V, respectively. Spa typing of the isolates showed MRSA isolates belonged to t304 and t6297. All the MRSA isolates were multidrug resistant. The MIC of oxacillin, vancomycin, ampicillin, ciprofloxacin and linezolid against MRSA isolates was 8->256 μg ml−1, 0.064–0.256 μg ml−1, 0.38–2 μg ml−1 and 6–8 μg ml−1, and 0.1 μg ml−1, respectively. The finding is worrying from public health point of view due to detection of MRSA and enterotoxin (seg,sei) producing S. aureus in milk samples which are commonly associated with human infection or food poisoning cases. This is the first comprehensive study with substantial numbers of samples for characterization of MRSA in bovine milk in India.
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•Occurrence of SCCmec type Iva and type V possessing MRSA in bovine milk in India.•Occurrence of t304 and t6297 spa types.•MRSA isolates were multidrug resistant.•MIC of anti-staphylococcal drugs against MRSA was detected.
Mastitis is a highly infectious disease prevalent in dairy cattle and it is majorly caused by Escherichia coli (E. coli). The objective of present study is to investigate the occurrence of virulence ...genes, antimicrobial susceptibility and comparative analysis of E. coli (IVRI KOL CP4 and CM IVRI KOL-1) isolates from mastitis infected animal. Whole-genome sequencing (WGS) was performed using a PacBio RS II system and de novo assembled using Hierarchical Genome Assembly Process (HGAP3). Bacterial Pan Genome Analysis Pipeline (BPGA) was used for pangenome analysis. A set of 50 E. coli isolates were used for comparative analysis (48 collected from the database and 2 reference sequences). Core genes were further concatenated for phylogenetic analyses. In silico analysis was performed for antibiotic resistance and virulence gene identification. Both of the E. coli isolates carried many resistance genes including, b-lactamase, quinolones, rifampicin, macrolide, aminoglycoside and phenicols resistance. We detected 39 virulence genes in IVRI KOL CP4 and 52 in CM IVRI KOL-1 which include toxins, adhesions, invasins, secretion machineries or iron acquisition system. High prevalence of mastitis strains belongs to phylogroups A, although few isolates were also assigned to phylogenetic groups B1 and B2. In conclusion, the present study reported the presence of genes involved in Adherence, Iron acquisition, secretion system and toxins which shown to be crucial in MPEC pathogenicity. This is the first whole genome analysis of MPEC strains to be carried out in Indian isolate to highlights the spread of resistance and virulence genes in food animals.
This study describes comparative occurrence and characterization of multidrug-resistant (MDR)
and
(KP) in healthy cattle (HC) and cattle with diarrhea (DC) in India. During 2018-2020, 72 MDR ...isolates, including 35
(DC: 27; HC 8) and 37
(DC: 34; HC: 3), from 251 rectal swabs (DC: 219; HC: 32) were investigated for extended-spectrum beta-lactamase (ESBL), AmpC type β-lactamase and carbapenemase production, antimicrobial susceptibility profile, biofilm production, and efflux pump activity. Fifty-five MDR isolates were ESBL producers (ESBLPs) (DC: 50; HC: 5) and ESBLPs from DC were coresistant to multiple antibiotics. The
CTX-M gene (50) was the most frequently detected β-lactamases followed by
AmpC (22),
TEM1 (13),
CMY-6 (6),
OXA1 (5),
PER (2),
DHA, and
FOX and
SHV12 (1 each). Plasmid-mediated quinolone resistance determinants
,
,
, and
were detected in 18, 16, 2, and 3 isolates, respectively. Twenty three isolates revealed mutation in
and
genes. Tetracycline-resistance markers
,
,
, and
were detected in 33, 10, 3, and 2 isolates, respectively. Only one of the 41 imipenem-resistant isolates harbored
NDM-5 and two were colistin-resistant. Altogether, 20 MDR isolates were strong biofilm producers and 19 harbored different virulence factors. This is the first ever report from India on the presence of MDR Enterobacteriaceae with resistance to even last-resort antimicrobials in the bovine diarrhea.
This study details about the phenotypic and molecular characteristics of multidrug‐resistant (MDR) Escherichia coli in the fresh community pond water (n = 257) collected from three districts of West ...Bengal, India. In total, 57 isolates were MDR of which 38 emerged as extended spectrum and 7 as AmpC‐type β‐lactamase producers in phenotypic assay. Among β‐lactamase genes, blaCTXM‐1was predominant (87.71%) followed by blaAmpC (77.2%) and blaTEM‐1 (22.8%). Six MDR strains carried metallo‐β‐lactamase (MBL, blaNDM‐1) gene. Tissue culture plate assay confirmed strong biofilm (SP) production in four MDR and one non‐MDR isolates. In PCR‐based replicon typing (PBRT), multiple plasmids of diverse replicon types (Frep, FIB, I1, FIA, K/B, HI1, and Y) were identified. The enterobacterial repetitive intergenic consensus–polymerase chain reaction (ERIC‐PCR)‐based phylogenetic analysis revealed a high degree of genetic divergence among the MDR isolates. Multiplex PCR‐based phylogrouping categorized 11 isolates as virulent (B2/D/F), which carried blaCTXM‐1 gene and three had blaNDM‐1 gene. Relative transcriptional activity of AcrAB efflux pump was significantly elevated among the SP and MBL producers. The presence of MDR E. coli isolates, particularly those resistant to carbapenem, in pond water used for daily domestic and household work, is a cause of concern as these pathogens may sneak into human food chain causing life‐threatening infections.
Practitioner Points
Multidrug‐resistant biofilm producing E. coli isolated from community pond water.
A few of them were carbapenem‐resistant and belonged to virulent (B2/D) types.
Expression of AcrAB efflux pumps was found significantly elevated among biofilm producers and carbapenem‐resistant population.
Fresh pond water harboured MDR, biofilm producing E. coli. Frequent resistance to tetracycline, cephalosporins and quinolone was noted. Majority were ESBL producers and a few belonged to virulent (B2/D) type.
The transmission of zoonotic bacteria through consumption of raw milk is complicated by the dissemination of antimicrobial-resistant bacteria. The present study was conducted to detect the occurrence ...of antimicrobial-resistant bacteria (ESBL-/AmpC-producing Klebsiella spp.) in cow’s milk originating from healthy or infected (mastitis) cattle in India. In total, 450 milk samples were collected from apparently healthy cattle and cattle suffering from clinical or sub-clinical mastitis. Out of 455 Klebsiella spp., 67 (14.73%) isolates were found to be ESBL producers in the double-disc diffusion test. The occurrence of ESBL-producing Klebsiella spp. was significantly (p < 0.05) higher in milk samples collected from cattle suffering with mastitis than in healthy cattle. Among the ESBL-producing Klebsiella spp., 56 (83.6%) isolates were also detected that produced AmpC β-lactamases. All the ESBL and AmpC-producing Klebsiella spp. possessed blaCTX-M (100%) and blaAmpC (100%), respectively. The present study revealed a higher occurrence of class 1 integron in ESBL-producing Klebsiella spp. isolates. All ESBL-producing- Klebsiella spp. isolates were multi-drug resistant. The ciprofloxacin- and/or levofloxacin-resistant Klebsiella spp. isolates possessed the quinolone resistance gene (qnrS). The co-trimoxazole-resistant isolates possessed the sul1 and sul2 genes. Phylogenetic analysis of the studied isolates revealed that strains isolated from the same location had a clonal relationship. The study increases consumer awareness of the need to avoid raw milk consumption to prevent the spread of antimicrobial resistance in the community.
The present investigation was carried out to study the vancomycin resistance pattern of Staphylococcus aureus isolates (n = 274) obtained from 352 milk samples of bovine (269) and caprine (63) ...clinical and subclinical mastitis from different districts of West Bengal, India. Of them, seven isolates (vancomycin-resistant S. aureus VRSA 1-7) exhibited resistance to vancomycin. Minimum inhibitory concentration of vancomycin (MIC
) for VRSA2 and VRSA3 was ≥16 μg/ml; thus categorized as VRSA. For rest of the isolates, MIC
was 8 μg/ml and they were grouped as vancomycin intermediate S. aureus (VISA). Even though all the isolates were resistant to cefoxitin and oxacillin and possessed mecA gene, none of them carried vancomycin resistance gene. Furthermore, all the seven isolates were subjected to Staphylococcal cassette chromosome mec (SCCmec) typing, Staphylococcal protein A (spa) typing, and enterobacterial repetitive intergenic consensus polymerase chain reaction. All the isolates except VRSA3 and VRSA4 from Kolkata district exhibited diverse genetic lineage, irrespective of their host and antibiotic resistance pattern. These two isolates showed clonal similarity (MRSA-SCCmec-V-spa t267) with methicillin-resistant S. aureus (MRSA) strains previously reported in human and animal infection. Isolation of VRSA and VISA could probably be due to intensive use of vancomycin in healthcare premises, which might have led to the development of glycopeptide-resistant strains and thereafter, further disseminated in the environment, including livestock farms. Detection of VRSA in milk is a serious concern as it may further cause health problems in the consumers. This is the first ever report of VRSA in food animals, even though the pathogen is otherwise prevalent in humans.
This study was undertaken to detect the prevalence of CTX-M-producing Klebsiella spp. in healthy broiler, indigenous, and kuroiler birds reared in West Bengal (India) during November 2014-February ...2015. In addition to CTX-M gene, the study was also conducted to reveal the occurrence of other β-lactamase and class I integron genes in Klebsiella spp. isolates along with their clonal relationship. A total of 321 cloacal swabs from healthy broiler, indigenous, and kuroiler birds were collected from different places of West Bengal, India. Klebsiella spp. isolation rate varies among different types of poultry birds (43.8-72.3%). In total, 33 (10.7%) Klebsiella spp. isolates were detected phenotypically as CTX-M producers and all the isolates possessed bla
in polymerase chain reaction. Whereas 17 (51.5%) and 16 (48.5%) Klebsiella spp. isolates possessed bla
, and bla
with bla
, respectively. None of the CTX-M-producing Klebsiella spp. isolates in this study possessed class I integron gene. Randomly amplified polymorphic DNA-based phylogenetic tree revealed the presence of clonal relationship among the CTX-M-producing Klebsiella spp. isolates, recovered from broilers and indigenous birds. This study identified broilers and indigenous game birds as a potential reservoir of CTX-M-producing Klebsiella spp., which could be transmitted to the human food chain directly or indirectly.
We investigated the occurrence of extended-spectrum β-lactamase (ESBL) and AmpC-type β-lactamase (ACBL) producing quinolone-resistant Klebsiella pneumoniae (KP) in milk samples of apparently healthy ...buffaloes (n = 348) and buffaloes (n = 19) with evidence of subclinical mastitis from seven districts of West Bengal, India. In total, 12 ESBL producing KP were isolated with blaCTX-M-15 gene and 7 of them were ACBL producers, as well. The blaCTX-M-15 genes were carried by transposable element ISEcp1. The plasmid-mediated quinolone resistance genes-qnrS, qnrA, qnrB, qepA, and aac(6')-Ib-cr were detected in five, one, three, four, and one isolate (s), respectively. In addition, eight isolates carried mutation in gyrase (gyrA) and six in topoisomerase IV (parC). Resistance markers/genes for sulfonamide (sul1), tetracycline tet(A) and tet(B), and aminoglycoside (aacC2) were also detected in eight, four, and one isolate(s), respectively. The class I integrons identified in five isolates carried aad2/aad5 and dfrA12/dfrA17 gene cassettes. The enterobacterial repetitive intergenic consensus-PCR revealed that all the isolates were genetically diverse and comprised a heterogeneous population. Isolation of multidrug-resistant KP, a typical nosocomial pathogen from buffalo milk, reiterates the need to monitor farm animals for ESBL producing Enterobacteriaceae and emphasizes on judicious use of antibiotics in animal husbandry sector.
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