Background: Nowadays, the emergence and dissemination of Extended-spectrum &beta;-lactamase (ESBL) and Carbapenemase producing Enterobacteriaceae (CPE) constitutes a major public health concern. The extensive and inappropriate use of antibiotics may lead to development of multidrug resistant (MDR) organisms. The antibiotic resistant genes are mostly situated conjugative plasmids, which are horizontally transmitted from one species to another in highly populous country like India.

Methods: Total 240 samples were collected from Ayodhya and Sultanpur districts of Awadh region of Uttar Pradesh (India). Confirmation of isolates was done by PCR analysis using species specific Uid A gene. ESBL and Carbapenemase producing isolates were confirmed using DDST, ESBL-E-strip and MBL E-strip test. Plasmid profile of these isolates was performed using GeneJet Plasmid Miniprep Kit and gel-electrophoresis.

Result: In this study total 186 (77.5%) isolates of E. coli were confirmed by PCR analysis. Total 130(54.16%) isolates were confirmed as ESBL and 10(4.16%) as Carbapenemase producer using E-ESBL strip and MBL-E strip test. All ESBL and Carbapenemase positive isolates were found 99-100% sensitive against chloramphenicol, polypeptides and aminoglycosides group of antibiotics and 80-100% resistance against 3rd, 4th group of cephalosporins, monobactam and penicillin group of antibiotics. Total 76.18% ESBL positive and 100% Carbapenemase positive isolates were found MDR. Plasmid profiling revealed total 63.84% ESBL positive isolates&nbsp;carried plasmid of molecular weight in range of 1.5 to &gt; 10kb with 1-3 number of bands and 80.0% Carbapenemase positive isolates carried plasmid with molecular weight 3 to &gt;10 kb with 1-3 number of bands.

Multidrug Resistance Pattern and Plasmid Profiling of ESBL and Carbapenemase Producing E. coli Isolates of Bovine Origin in Awadh Region of Uttar Pradesh

<div>Background: Antibiotic resistance is an emerging concern in the therapy of clinical infections worldwide. Previous studies conducted in our laboratory have confirmed an increase in the prevalence of extended spectrum beta-lactamase (ESBL) among the Gram-negative bacterial pathogens associated with dogs, which could act as a potential source for the transfer of these resistant pathogens or their genetic determinants to human. Since carbapenems are the last resort drugs against these resistant pathogens, the study was aimed to isolate and characterise carbapenem resistance among Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae) and Pseudomonas aeruginosa (P. aeruginosa) associated with common clinical infections in dogs. 
 
Methods: A total of 100 samples were collected from lesional skin, urine and anterior vagina of dogs presented to the Veterinary Hospitals of Kerala Veterinary and Animal Sciences University at Mannuthy and Thrissur. The samples were cultured onto Brain Heart Infusion Agar (BHIA), Eosin Methylene Blue (EMB) and Mac Conkey (MAC) for isolation of bacteria. Identification of the isolates was performed based on cultural, morphological and biochemical characteristics. The isolates were subjected to antimicrobial susceptibility test (ABST) against the 12 commonly used beta-lactam and non&ndash;beta-lactam group of antibiotics by disc diffusion method and further subjected to screening for ESBL double disc diffusion method. Carbapenem-resistant isolates were subjected to phenotypic confirmatory test for carbapenemase production employing Imipenem-EDTA and Ertapenem-boronic acid minimum inhibitory concentration (MIC) strip method. 
 
Result: Forty four Gram-negative bacterial isolates obtained were viz., E. coli (30), K. pneumonia (11) and P. aeruginosa (3) from the 100 samples. Apart from these, other isolates obtained were Staphylococcus spp. (53) and Bacillus spp. (2). All the Gram-negative isolates were subjected to ABST employing 12 common antibiotics belonging to beta-lactam and non-beta-lactam groups. Multidrug resistance (MDR) could be observed in 28 E. coli, 11 K. pneumoniae and three P. aeruginosa isolates. All the 42 MDR isolates showed positive results for ESBL production. A total of 14 isolates out of the 44 Gram-negative bacilli were found to be resistant to carbapenem either to imipenem, meropenem or ertapenem. Among the 14 Gram-negative isolates, nine turned out to be positive for metallo-beta-lactamase (MBL) and none for K. penumoniae carbapenemase (KPC) on phenotypic confirmatory test for detecting major carbapenemase enzymes. The present study documented that Gram- negative bacteria like E. coli, K. pneumoniae and P. aeruginosa isolated from dogs are showing an increase rate of resistance against carbapenems which are the last resort drugs against ESBL producers. Hence, there is an urgent need to curb the irrational and excessive use of antibiotics in veterinary sector.</div>

<div>Phenotypic Characterisation of Klebseilla pneumoniae Carbapenemase and Metallo Beta-lactamase in Carbapenem Resistant Gram-negative Bacteria</div>

Background: Carbapenems are used as last resort of antibiotics to treat infection caused by multi-drug resistant pathogens of human, hence resulting into emergence and dissemination of Carbapenem-resistant Enterobacteriaceae (CRE). The occurrence of CRE among livestock is of great concern as these can be transferred to human and animals through adulterated food, water and environment. Therefore a specific study on the occurrence of CRE and their AMR profiling is warranted with regard to therapeutic options. 
 
Methods: Total 240 faecal samples were collected from Ayodhya and Sultanpur districts of Awadh region of Uttar Pradesh (India). The E. coli and Klebsiella spp. isolates were confirmed by PCR analysis using species specific uidA and 16S rRNA genes respectively. Carbapenemase producing isolates were confirmed by DDST, MBL E-strip and PCR analysis by targeting bla-NDM, bla-KPC and bla-OXA-48 genes. Antibiotic resistance profiling was performed against 20 antibiotics of 12 different classes. 
 
Result: In this study, PCR analysis revealed 90.80% confirmed isolates including 71.67% E. coli and 19.16% Klebsiella spp., out of which 36/218 (16.51%) and 28/218 (12.84%) isolates were confirmed as carbapenemase producer by DDST and MBL E-strip test respectively. Carbapenemase genes were detected in 17/218 (7.80%) isolates and among them bla-KPC gene was detected in both isolates while bla-NDM and bla-OXA-48 were detected only in E. coli and Klebsiella spp. respectively. All phenotypically confirmed carbapenemase positive isolates of E. coli and Klebsiella spp. were found 100% sensitive to aminoglycosides, polypeptides and 85-100% resistance against carbapenem, 3rd and 4th&nbsp;generation cephalosporines, monobactams and penicillin class of antibiotics. 
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Molecular Characterization and Antimicrobial Resistance Profiling of Carbapenemase Producing E. coli and Klebsiella spp. Isolates of&nbsp; Bovine Origin in Awadh Region of Uttar Pradesh

<div>Background: Among enterobacteria E. coli and Klebsiella spp. are of great concern in health care settings, as these bacteria sometimes may contaminate the milk due to unhygienic practices and poor udder condition which have been associated with various illnesses. Therefore, this study aimed to detect the carbapenem resistant E. coli and Klebsiella spp. of bovine milk origin with regard to the risk of human transfer via the food chain in community. 
 
Methods: Total 240 samples were collected from Ayodhya and Sultanpur districts of Eastern Plain Zone of Uttar Pradesh (India). Confirmation of E. coli and Klebsiella spp. isolates was done by using species specific uidA and 16S rRNA gene, respectively. Then, carbapenemase positive E. coli and Klebsiella spp. were confirmend by DDST, MBL E-strip test and PCR analysis by targeting (bla-NDM, bla-OXA-48 and bla-KPC). Antibiogram of all carbapenemase positive isolates was performed against 20 antibiotics of 12 different classes. 
 
Result: In the present study, total 74(30.83%) isolates were identified including 55(22.92%) E. coli and 19(7.92%) Klebsiella spp. by PCR, out of which 12(16.21%) isolates were confirmed as carbapenemase producers comprising 7(12.72%) E. coli and 5(26.31%) Klebsiella spp by DDST and E-strip. All carbapenemase positive E. coli were found 100% sensitive to polymyxin-B and chloramphenicol, while all Klebsiella spp. were 100% sensitive to amikacin and polymyxin-B. Resistance against imipenem, meropenem, cefotaxime, cefpodoxime, ceftazidime, ceftriazone, aztreonam and ampicillin ranged between 80.0%-100%. All carbapenemase positive isolates were found multidrug resistant. Carbapenemase genes bla-NDM and bla-KPC were detected in E. coli while bla-OXA-48 and bla-KPC were detected in Klebsiella spp.</div>

​Detection of Carbapenemase Genes (bla-NDM, bla-KPC, bla-OXA-48) in Escherichia coli and Klebsiella species, Isolated from Milk Samples of Bovine in Eastern Plain Zone of Uttar Pradesh

<div>Background: Carbapenems are beta-lactam antibiotics that are considered as the last line of therapy against multidrug resistant extended spectrum beta-lactamase. The resistance to carbapenems predominantly through carbapenemase is one of the most important emerging health problems worldwide in the therapy of clinical infections. The objective of the present study is to determine the presence of carbapenemase encoding genes among Gram- negative bacterial spp. associated with clinical infections in dogs.&nbsp; 
 
Methods: 30 Escherichia coli, 11 Klebsiella pneumoniae and three Pseudomonas aeruginosa isolated from urine, swabs from lesional skin and anterior vagina of dogs presented with different clinical ailments formed the samples for the study. Polymerase chain reaction was carried out to detect the presence of carbapenemase encoding genes viz., KPC, NDM, OXA, VIM and IMP among the isolates. 
 
Result: Out of the 44 Gram- negative isolates tested, 28 (76.3%) were positive for at least one tested carbapenemase gene. The highest frequency of carbapenemase recorded was for NDM followed by OXA-181, KPC, OXA-48 and VIM. Our study identified a high prevalence of carbapenemases among companion animals like dogs which could act as potential source of transmission of these resistance bacteria or their genomes to humans.</div>

<div>Molecular Detection of Carbapenem Resistant Gram Negative Bacterial Isolates from Dogs&nbsp;</div>

<div>Background: The emergence of a multidrug resistance phenotype is most likely outcome of co-existing multiple mechanisms. The underlying molecular mechanism of carbapenem resistance is poorly understood in bacterial isolates of animal origin. The present study investigates the resistance mechanism for reduced susceptibility to carbapenem in E. coli. 
&nbsp;&nbsp; 
Methods: The study includes 12 multidrug resistant E.coli isolates showing decreased susceptibility to imipenem. Carbapenemase production was evaluated by the mCarba-NP test. The presence of the carbapenem resistance gene was confirmed by multiplex PCR. Efflux pump activity was accessed by broth microdilution method with or without selective efflux pump inhibitor. We analyzed differential expression of the outer membrane porin genes (ompF, ompC) and efflux pump genes (acrA, acrB) by quantitative real-time PCR analysis.&nbsp; 
 
Result: We observed increased expression of acrA and acrB genes in all the E. coli isolates under imipenem stress. A positive correlation (p&lt;0.001) was evident between imipenem MIC and acrA and acrB over-expression in these isolates. A significant decrease in ompF and ompC expression was observed in these isolates under imipenem stress. The change in MIC was not correlated with the corresponding OmpC and OmpF gene expression. The results suggest the importance of efflux pump and porins proteins in the emergence of multidrug resistance besides carbapenemase production.</div>

Imipenem Induced Transcriptional Modulation of Multidrug Efflux Pumps and Porin Genes in Carbapenem Resistant Escherichia coli Isolates of Animal Origin

Multidrug Resistance Pattern and Plasmid Profiling of ESBL and Carbapenemase Producing E. coli Isolates of Bovine Origin in Awadh Region of Uttar Pradesh

Phenotypic Characterisation of Klebseilla pneumoniae Carbapenemase and Metallo Beta-lactamase in Carbapenem Resistant Gram-negative Bacteria

Molecular Characterization and Antimicrobial Resistance Profiling of Carbapenemase Producing E. coli and Klebsiella spp. Isolates of Bovine Origin in Awadh Region of Uttar Pradesh

Detection of Carbapenemase Genes (bla_-NDM, bla_-KPC, bla_-OXA-48) in Escherichia coli and Klebsiella species, Isolated from Milk Samples of Bovine in Eastern Plain Zone of Uttar Pradesh

Molecular Detection of Carbapenem Resistant Gram Negative Bacterial Isolates from Dogs

Imipenem Induced Transcriptional Modulation of Multidrug Efflux Pumps and Porin Genes in Carbapenem Resistant Escherichia coli Isolates of Animal Origin

Multidrug Resistance Pattern and Plasmid Profiling of ESBL and Carbapenemase Producing E. coli Isolates of Bovine Origin in Awadh Region of Uttar Pradesh

Phenotypic Characterisation of Klebseilla pneumoniae Carbapenemase and Metallo Beta-lactamase in Carbapenem Resistant Gram-negative Bacteria

Molecular Characterization and Antimicrobial Resistance Profiling of Carbapenemase Producing E. coli and Klebsiella spp. Isolates of Bovine Origin in Awadh Region of Uttar Pradesh

​Detection of Carbapenemase Genes (bla-NDM, bla-KPC, bla-OXA-48) in Escherichia coli and Klebsiella species, Isolated from Milk Samples of Bovine in Eastern Plain Zone of Uttar Pradesh

Molecular Detection of Carbapenem Resistant Gram Negative Bacterial Isolates from Dogs

Imipenem Induced Transcriptional Modulation of Multidrug Efflux Pumps and Porin Genes in Carbapenem Resistant Escherichia coli Isolates of Animal Origin

Detection of Carbapenemase Genes (bla_-NDM, bla_-KPC, bla_-OXA-48) in Escherichia coli and Klebsiella species, Isolated from Milk Samples of Bovine in Eastern Plain Zone of Uttar Pradesh