<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>

Background: The orexin (OX) is a hypothalamic neuropeptide that exists in two forms, orexin A (OXA) and orexin B (OXB), which bind two receptors (OX1 and OX2). While orexin B shows selectivity for OX2, OXA is potent at both receptors. Orexins are also reported in other mammalian organs where they undergo many regulatory functions. However, their detection in the dromedary camel reproductive organs remains to be revealed. 
 
Methods: We investigated the immunoreactivity of orexin OXA and OX1 in the testis of young (6-12 month), pre-pubertal (2-3 years) and adult (6-15 years) dromedary camels. We also studied the relative expression of hypocretin neuropeptide precursor (HCRT) in the testicular tissue of these age groups. 
 
Result: Positive immunoreactions of OXA and OX1 were shown in both the tubules and interstitium of dromedary camel testis in the young animals. While positive OXA and OX1 immune-labeling was detected in the spermatogonia, Sertoli cells and Leydig cells in all ages, adult camels also showed positive OXA and OX1 signals in the primary spermatocytes, spermatozoa and interstitial blood vessels. In testes at pre-pubertal and adult stages, spermatogenic cells, Sertoli cells and Leydig cells showed stronger OXA and OX1 reactions than in young camels. Real time PCR analysis also revealed that HCRT was expressed in the testis of dromedary camels with minimal expression in young camels. In conclusion, the current study provides the first evidence for the presence of OXA and OX1 in the testis of adult and younger dromedary camels. Before puberty, the binding of OXA and OX1 in the camel testis might be important for development and proliferation of spermatogenic cells, Sertoli cells and Leydig cells; in adult animals they may play a role in steroidogenesis and spermatogenesis.

Age-related Localization and Expression of Orexin A and its Receptor 1 in Dromedary Camel Testis&nbsp;

Background: The use of antimicrobials in food animals significantly contributes to the development of antimicrobial resistance in nature. This study aimed to determine the prevalence of ESBL-producing E. coli isolated from cattle, buffalo, sheep and goat fecal samples.&nbsp; 
 
Methods: In this study out of 2025 healthy animals from 10 different farms, fecal samples were randomly collected by rectal swabs from 200 animals (50 each from cattle, buffalo, sheep and goat). E. coli was isolated and further studied for AMR and ESBL production by phenotypic assays and PCR targeting ESBL genes.&nbsp; 
 
Result: The overall prevalence of E. coli was 81.50% with the highest isolation rate from cattle and goat samples. Out of 163 isolates, 45.39% were ESBL-producers. Genes encoding beta-lactam resistance viz. blaSHV, blaTEM, blaCTX-M and blaOXA were detected in 44.59% ESBL-positive isolates. The blaSHV was present in only cattle isolates, while blaCTX-M was present in E. coli isolated from all animals. The prevalence of beta-lactam genes was found as blaSHV (2.70%), blaTEM (12.16%), blaCTX-M (22.97%) and blaOXA (6.75%). A high degree of resistance to multiple antimicrobials was observed. Maximum susceptibility of E. coli was observed for trimethoprim, imipenem and chloramphenicol. This study demonstrated that food animals could be the source of ESBL-producing multidrug-resistant E. coli. The present findings are significant in the context of AMR monitoring in India.

Phenotypic and Molecular Characterization of Escherichia coli Isolated from Food Animals for Antimicrobial Resistance&nbsp;

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. 
&nbsp;

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 style="text-align:justify">The present study was carried out to characterize beta-lactam resistance in Escherichia coli isolated from healthy and diarrhoeic dogs. A total of 93 E. coli were isolated from the rectal swabs of 136 dogs (60/92 of healthy dogs and 33/44 of diarrhoeic dogs). Predominant serotypes detected include rough (19 isolates), O141 (5), O9 (2), O126 (2), O128 (2), O15, O20, O35, O49, O63, O85, O101, O116, O117, O118, O119 (1 isolate each) and the rest of 52 isolates were untypable (UT). Disc diffusion method revealed resistance to cefotaxime (41.9%), ceftriaxone (34.4%), ceftazidime (30.1%) and aztreonam (18.2%). Overall frequency of extended spectrum beta-lactamase (ESBL) phenotype was found to be 29% (27/93). Beta-lactamase genes detected include blaAmpC (86.0%), blaSHV (30.1%), blaCTX-M group-1 (19.3%), blaTEM (17.2%), blaOXA (13.9%) and blaCTX-M group-2 (7.5%). The study revealed resistance to commonly prescribed beta-lactams, with ESBL phenotype in E. coli of canine origin in Andhra Pradesh, India.&nbsp;</div>

<div style="text-align: justify;">Occurrence of beta-lactamase producing Escherichia coli in healthy and diarrhoeic dogs in Andhra Pradesh, India</div>

<div>Background: Proteus mirabilis is one of the organisms which is often associated with urinary tract infections of humans as well as animals. As a member of Enterobacteriaceae family, the level of antimicrobial resistance tend to pose a significant public health risk. Hence, the present study was undertaken to study antibiogram profiles, multidrug resistance P. mirabilis isolates and detection of &beta;-lactamase activity in them. 
 
Methods: A total of 175 P. mirabilis isolates from different sources were subjected to antibiotic sensitivity/ resistant test by Kirby Bauer disc diffusion method. Detection of ESBL production was done phenotypically by Phenotypic Screening Test and Phenotypic Confirmation Test as recommended by CLSI guidelines and genotypically using multiplex PCR assay to detect different classes of &beta;-lactamase genes. This study was carried out from March 2017 to August 2018 in and around areas of Krishna District Andhra Pradesh. 
 
Result: Out of 175 P. mirabilis isolates screened, antibiogram revealed highest sensitivity towards gentamicin (76.57%), followed by ampicillin (64.57%), kanamycin (61.14%), amikacin (60.57%) and streptomycin (43.42%). Higher resistance was observed for erythromycin (71.42%), nalidixic acid (62.85%), ciprofloxacin (62.85%), tetracycline (60%), polymyxin-B (60%), cefoxitin (49.14%) and amikacin (36%). &beta;-lactamase genes were detected in a total of 23 isolates (13.14%). Prevalence rates of &beta;-lactamase genes among different samples was 23.6%, 11.1%, 10.8% and 42.8% from chicken, pork, poultry cloacal swabs and human urine samples, respectively with blaTEM being the predominant gene detected (69.56%) followed by blaOXA (26.08%), blaAmpC gene FOX (13.04%), blaCTX-M group I (4.34%), blaSHV (4.34%) and blaAmpC gene CIT (4.34%) among all the tested P. mirabilis isolates.</div>

<div>A Study on Antibiogram and Beta-lactam Resistance of Proteus mirabilis Isolated from Animals and Humans in Andhra Pradesh, India</div>

Background: Multidrug-resistant pathogens are a significant cause of morbidity globally. The recent rise in resistant bacterial strains is largely attributed to the extensive use of antibiotics in human and animal treatment. In recent years, there has been a notable increase in drug-resistant Klebsiella pneumoniae strains, posing a major challenge for clinicians worldwide. 
 
Methods: In the current study, 121 samples were obtained from bovine mastitis milk and faeces, dairy cattle farm premises, chicken cloacal swabs, poultry farm litter and water. The samples were processed for the isolation and identification of Klebsiella species. Klebsiella isolates were identified by cultural, morphological and biochemical characteristics and were subsequently confirmed by PCR assays that targeted species-specific genes. The isolates were subjected to in vitro antibiotic sensitivity test. The DNA from all the isolates was extracted and subjected to PCR to identify the presence of 10 different Antibiotic Resistance Genes (ARG) using published primers. The ARGs targeted were blaTEM, blaSHV, blaOXA, blaIMP, DHAM, MOXM, tetA, tetB, sul1 and aadA. 
 
Result: Out of 121 samples, twenty-six (21.4%) samples were identified as Klebsiella pnuemoniae isolates. The occurence of Klebsiella pnuemoniae in samples from different sources was- 3/33 (09%) from mastitis milk, 8/27 (29.6%) from faecal samples and 04/14 (28.5%) from environmental samples collected from dairy farms. Similarly, Klebsiella pnuemoniae prevalence was recorded to be 6/33 (18.1%) in cloacal swabs and 5/14 (35.7%) from environmental samples collected from poultry farms. On in vitro antibiotic sensitivity test, all the Klebsiella pneumoniae isolates were found to be resistant to penicillin, amoxicillin, cefuroxime and ampicillin/ sulbactam (100% each), followed by cefalexin (65.3%), streptomycin (53.8%), tetracycline (38.4%), imipenum and co-trimoxazole (34.6% each), ofloxacin (30.7%), norfloxacin (27%), ceftriaxone (23%), amikacin (15.38%) and gentamicin (11.5%). All the K. pneumoniae isolates were found to be Multi Drug Resistant (MDR). Out of 26 isolates, 25 (96%) were positive for Sul1, 22 (84.6%) for blaTEM, 19 (73%) for blaSHV, 19 (3%) for tetA, 16 (61.5%) for aadA, 10 (38.4%) for OXA, 9 (34.6%) for tetB and 2 (7.6%) for DHAM and MOXM each. None of the isolates harboured blaIMP gene.

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

Age-related Localization and Expression of Orexin A and its Receptor 1 in Dromedary Camel Testis

Phenotypic and Molecular Characterization of Escherichia coli Isolated from Food Animals for Antimicrobial Resistance

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

Occurrence of beta-lactamase producing Escherichia coli in healthy and diarrhoeic dogs in Andhra Pradesh, India

A Study on Antibiogram and Beta-lactam Resistance of Proteus mirabilis Isolated from Animals and Humans in Andhra Pradesh, India

Multi Drug Resistant (MDR) Klebsiella pnuemoniae Isolated from Different Sources