The study was undertaken to investigate the occurrence and to characterize the ESBL/beta-lactamase producing-Escherichia coli in healthy pigs of organized and backyard farms in West Bengal, India. ...Total 200 rectal swabs were collected randomly from healthy pigs maintained in four organized farms and 10 backyard farms (n=100 each) and 76 isolates were identified as E. coli from organized (48/100, 48%) and backyard pigs (28/100, 28%). Twelve E. coli isolates (6%) in the present study were detected to possess any of the ESBL/beta-lactamase genes studied. ESBL/beta-lactamase producers were isolated with significantly more frequency from backyard pigs than the organized farm pigs (p=0.026). Six of ESBL/beta-lactamase producing isolates were phenotypically confirmed as CTX-M producers and ten of them were confirmed as TEM/SHV producers. PCR and sequencing of the amplified product from representative isolates revealed the presence of blaCTX-M-9, blaSHV-12 and blaTEM-1. No unique combination of the studied beta lactamase genes for organized and backyard farm pig isolates was noted. The ESBL isolates belonged to O13, O55, O133, O153, O157, O158, O166, rough and OUT serogroups. The association of heat labile toxin (elt) (p<0.0005) with organized farm isolates and heat stable toxin (estA) (p=0.0143) with backyard piggery sector was significantly higher. The ESBL/beta-lactamase producers from organized farm (Ak/Ex) and indigenous pigs (Ak/Ex/Te; Ak/CoT/G) showed a characteristic phenotypical antibiotic resistance pattern. Two pairs of isolates from organized and backyard farm pigs showed clonal relationship indicating a possible transmission between the farms which were situated adjacently. Thus the present study revealed backyard farm pigs as major source of ESBL/beta-lactamase producing-E. coli associated with STa and characteristic antibiotic resistance pattern in India.
•Higher carriage of ESBL/beta-lactamase producing E. coli in backyard farm pigs than the organized farm pigs•Positive correlation between the carriage of ESBL genes and ETEC toxin genes•No specific bla profile was detected in the isolates of both types of pigs but characteristic phenotypical resistance pattern•Clonal relationship between the ESBL producers of backyard and organized farm pigs•Isolation of O157 serogroup and association of STa with the E. coli isolates of indigenous pigs make them potential public health hazard•First report of such a comparative study
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.
<正>Objective:To evaluate the antimicrobial efficacy of berberine,a plant alkaloid.Methods:Five multi-drug resistant（MDR） STEC/EPEC and five MDR ETEC isolates from yaks with haemorrhagic ...diarrhoea were selected for the study.Antibacterial activity of berberine was evaluated by broth dilution and disc diffusion methods.The binding kinetics of berberine to DNA and protein was also enumerated.Results:For both categories of enterovirulent Escherichia coli（E.roli） isolates, berberine displayed the antibaclerial effect in a dose dependent manner.The MIC50 of berberine chloride for STEC/EPEC isolates varied from 2.07μM to 3.6μM with a mean of（2.95±0.33）μM where as for ETEC strains it varied from 1.75 to 1.96μM with a mean of（1.87±0.03）μM. Berberine bind more tightly with double helix DNA with Bmax and Kd of（24.68±2.62） and（357.8±57.8）,respectively.Berberine reacted with protein in comparatively loose manner with Bmax and Kd of（18.9±3.83） and <286.2±113.6),respectively.Conclusions:The results indicate clearly that berberine may serve as a good antibacterial against multi drug resistant E.coli.
Persistence of antibacterial drugs for prolonged period in milk increases the probability of antimicrobial resistance progress. Ceftizoxime was found to be excreted in milk for a prolonged period in ...goats, cows and buffaloes following intravenous injection of ceftriaxone and ceftizoxime. A single dose of ceftriaxone was administered intravenously in healthy control goats (group I) and a single oral dose of the commercial mammary protective polyherbal drug (1.9 gm) was given one hour prior to intravenous ceftriaxone injection in healthy (group II) and induced mastitic (group III) goats to evaluate milk disposition of ceftizoxime following single intravenous dosing of ceftriaxone at 42.25 mg kg
.Ceftriaxone/ceftizoxime was analyzed by HPLC. The t
α and t
β values were 14.755 ± 2.733 and 149.079 ± 18.565 hour, respectively indicating prolonged persistence of ceftizoxime in milk. The polyherbal drug increased the milk concentration at later hours and hastened the excretion of ceftizoxime from milk compared to control group. Ceftriaxone could not be detected in milk. The study suggested that adjunct single or repeated therapy of the polyherbal drug may cause non persistence of ceftriaxone and shorter persistence of ceftizoxime in milk.
•We isolated 18 ESBL producing MDR E. coli from cattle and poultry.•Isolates carried β-lactamase, PMQR, sul and ExPEC virulence genes.•Class-1 integron analysis revealed aadA2 and dfrA12 gene ...cassettes.•Phylogenetic analysis reflected genomic diversity among the isolates.•This is the first ever report of this kind from India.
The present study was undertaken to determine the occurrence and characterization of extended spectrum beta-lactamase (ESBL) producing Escherichia coli isolated from cattle and poultry in Odisha, India. Of 316 E. coli isolated from 305 samples (170 fecal samples from poultry and 135 milk samples from cattle), a total of 18 E. coli isolates were confirmed as ESBL producers by combination disc method and ESBL E-test. The isolates were resistant to oxyimino cephalosporins and monobactam as revealed by disc diffusion assay and determination of minimum inhibitory concentration. Resistance against other antibiotics was frequently noted as well. Further, beta-lactamase genes viz., blaSHV, blaCTXM, blaTEM and blaampC were detected in 17, 13, 9 and 2 isolates, respectively in PCR. Of the 18 ESBL strains, 16 were positive for class I integron (int1), nine of them carried sulphonamide resistance gene (sul1) and one harbored quinolone resistance gene (qnrB). Virulence markers for extraintestinal pathogenic E. coli like astA, tsh and iucD were also present in 4, 3 and 3 isolates, respectively. All the PCR amplified products were cloned and subjected to sequencing for homology analysis and data were submitted to gene bank. Sequence analysis of the amplified variable regions of class 1 integron of four representative isolates revealed the presence of aadA2 and dfrA12 gene cassettes conferring resistance to aminoglycosides and trimethoprim, respectively. Most of the ESBL producing strains emerged as single lineage through phylogenetic analysis by RAPD and ERIC PCR. This is the first ever systemic study on multidrug resistant ESBL producing E. coli in food producing animals from India.
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.
The present study was undertaken to detect the occurrence of beta-lactamase-/AmpC-producing
Klebsiella
and
Escherichia coli
in healthy pigs, feed, drinking water, and pen floor or surface soil. The ...study also intended to detect the clonal relationship between the environmental and porcine isolates to confirm the route of transmission. Rectal swabs and environmental samples were collected from apparently healthy pigs kept in organized or backyard farms in India. The pigs had no history of antibiotic intake. Production of phenotypical beta-lactamase, associated genes, and class I integron gene was detected in
E. coli
and
Klebsiella
isolates. The phylogenetic relationship among the isolates was established on the basis of Random amplification of polymorphic DNA banding pattern. Beta-lactamase-producing
Klebsiella
were isolated from healthy pigs (20.0%), pen floor swabs/surface soil swabs (14.0%), and drinking water (100%).
Escherichia coli
isolated from healthy pigs (14.4%), pen floor/surface soil (8.0%), and drinking water (33.3%) were detected as beta-lactamase producers. Majority of beta-lactamase-producing isolates possessed
bla
CTX-M-9
. Further, 35 (81%)
Klebsiella
and all the
E. coli
isolates were detected as AmpC beta-lactamase ACBL producers and possessed
bla
AmpC
. Sixteen beta-lactamase-producing
Klebsiella
(37.20%) and 13
E. coli
(86.67%) possessed class I integron. Few resistant isolates from environmental sources (surface soil swab and drinking water) and the studied pigs were detected within the same cluster of the dendrogram representing their similarities. The study indicated about the possible role of contaminated environment as a source of beta-lactamase/AmpC-producing
Klebsiella
and
E. coli
in pigs.
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