<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: We investigated the occurrence of blaCTX-M carrying extended spectrum beta lactamase (ESBL) producing Escherichia coli in pigs from 8 North-eastern states of India with special emphasis on the transferability of ESBL gene from resistant E. coli strains to the susceptible Salmonella strains by in vitro and in vivo. 
 
Methods: Fecal samples (n=790) were collected from pigs reared under organized and unorganized farming set up of entire North-eastern region of India. All the samples were processed for isolation and identification of E. coli. All the isolates were subjected to antimicrobial sensitivity assay by disc diffusion method followed by determination of ESBLs producing ability by double disc synergy test (DDST). All the ESBLs producing isolates were screened for blaCTX-M gene by PCR using specific primers. The representative blaCTX-M gene positive isolates were used as donor to determine the ability to transfer of resistance gene in Salmonella by in vitro and in vivo assays with and without antibiotic selection pressure. &nbsp; 
 
Result: A total of 2,291 E. coli was isolated, of which 1113 and 1178 were from organized and unorganized farms, respectively. Majority of the isolates were multi-drug resistant with highest resistance against amoxicillin (84.81%) followed by cefalexin (77.17%), sulphafurazole (56.79%), piperacillin (46.40%), tetracycline (38.29%) and cefexime (35.66%). Isolates from unorganized farms showed higher resistance than the isolates recovered from organized farms. A total of 654 (28.55%) isolates were confirmed as ESBL producers by double disc synergy test (DDST) method, of which 65 (2.84%) isolates were positive for blaCTX-M gene. Genotypically, isolates with specific amino acids substitution revealed variation in their antibiotic susceptibility by phenotypic method. blaCTX-M gene could be successfully transferred horizontally from E. coli (donor) to Salmonella (recipient) by in vitro (3.6&plusmn;2.07x10-8 to 4.4&plusmn;2.88x10-8 transconjugate per donor) and in vivo method. By in vivo method in pig model, the frequency of transfer was higher under the antibiotic selection pressure (6.6&plusmn;3.05x10-5 to 7.2&plusmn;1.92x10-5 trans-conjugants per donor) than without antibiotic pressure (5.6&plusmn;2.3x10-4 to 6.8&plusmn;3.35x10-4 trans-conjugants per donor).</div>

Transferable blaCTX-M Carrying Multidrug Resistant Escherichia coli from Pig Population of North Eastern Region of India

<div>Background: The current situation of multidrug resistant bacteria has become a worrisome issue in occurrence of disease outbreak. Multidrugresistant E. coli can be readily encountered in farms during daily clinical practice and veterinarian should act timely. Over past many reported increased incidences of Extended-Spectrum Beta-Lactamase (ESBL)-producing E. coli. The introduction of ESBL-producing E. coli from poultry farms to the environment may pose a health risk if these bacteria reach places where people may become exposed and we report that different methods of identifications.&nbsp; 
 
Methods: 60 fecal samples were collected from apparently healthy and infected poultry in the mixed age group. Bacterial isolates were identified on colony morphology and biochemical properties. ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the TEM, SHV and CTX-M genes. 
 
Result: In the present study, 40 isolates of&nbsp;E.coli were isolated from 60 faecal samples. Out of 40 isolates, 12 isolates were resistant to two or more than two antibioticsand were positive for ESBL by combination disc test. CTX-M genes were detected in 6 (50%) isolates. TEM and SHV were not detected in any of the isolates. This study showed high resistance of E. coli to antibiotics, particularly in the third generation cephalosporins but were more susceptible to Gentamicin, Aztreonarm, Cefrtrazindime and Cefotoxime. Laboratory monitoring and detection of E. coli of ESBL producing bacteria important steps in the appropriate treatment for farm based poultry industry and infection control efforts.</div>

Phenotypic Characterization and bla Gene Detection in Antibiotic Resistant Escherichia coli Obtained from Poultry Fecal Samples

<div>Background: Higher prevalence of ESBL producers is alarming in dairy sector. So limiting antimicrobial use may curtail the selection and persistence of predominant ESBL and conjugative plasmids among strains. The study was aimed to determine the occurrence of extended-spectrum beta-lactamase (ESBL) producing E. coli as well as their genetic diversity, antimicrobial resistance and integrons in bovines.&nbsp; 
 
Methods:&nbsp;A total of 180 faecal samples were screened for the presumptive ESBL producing E. coli isolates. All ESBL producing isolates were subjected to the screening test and were confirmed as ESBL producers by double disc. Further, these ESBL producers were tested for the presence of bla genes and those found positive were tested for multidrug resistance (MDR) by disk diffsion. MDR positive isolates were further tested for the presence of intI 1, intI 2 and intI 3 genes. 
 
Result: A total of 360 presumptive ESBL producing E. coli isolates were obtained. Out of which, 154 (42.77%) isolates were found to be resistant and were confirmed as ESBL producers. Of 154 ESBL isolates, 120 (77.92%) isolates carried the gene/s screened for blaTEM, blaCTX-M and blaSHV genes are were declared as multi drug resistant. Out of 120 MDR isolates, 59 tested positive for the integron gene.</div>

Molecular Characterization and Antimicrobial Resistance Profiling of Extended Spectrum Beta-lactamase (ESBL) Producing Escherichia coli in Bovines from J and K, India

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: Cephalosporins are major antimicrobials used to treat serious infections. However, their effectiveness is being compromised by the emergence of Extended Spectrum Beta Lactamases (ESBL). The study was aimed to perform plasmid profiling of ESBL producing Escherichia coli from diarrheic calves. 
 
Methods: A total of 104 rectal swabs were collected from diarrheic calves (Less than 1 year old) from dairy farms (buffalo calves) and gaushalas (cattle) in and around Jabalpur (M.P.). Total of&nbsp; 81 E. coli isolates were isolated. The ESBL screening being conducted by using cefpodoxime, ceftazidime, cefepime and aztreonam antibiotics for initial screening. The phenotypic confirmation of the ESBL producing strains was performed through ESBL phenotypic identification kit and Double Disc Diffusion Test (DDDT). While, for genotypic detection PCR was performed for bla TEM, bla CTX-M and bla AmpC. Plasmid profiling of resistant isolates was performed. 
 
Result: Out of the 81 E. coli isolated, 07 (8.64%) were phenotypically confirmed as ESBL strains. PCR amplification revealed 03 (60%) isolates positive for bla TEM (867bp) and one (20%) isolate as bla CTX-M (540bp) genes, respectively. bla AmpC gene was not detected from these isolates. Co expression of bla TEM and bla CTX-M was recorded in only one diarrheic buffalo calf. Plasmid profiling of the isolates showed that all the isolates were harboring plasmids of varying size and number. The plasmid number among the isolates ranged from 3-4 plasmids per isolate and plasmid size varied from 900bp to &gt;10kb. 
&nbsp;

Detection and Plasmid Profiling of Extended Spectrum Beta Lactamase Producing Escherichia coli from Diarrheic Calves

<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: 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>Background: Antimicrobial resistance (AMR) due to emergence and spread of Extended-spectrum &beta;-lactamase (ESBL) producing bacteria are point of discussion in both human as well as in animals across the world as it is one of the latest challenges faced by scientific community. The present study highlighted the prevalence of ESBL producing E. coli and Klebsiella spp. isolated from buffaloes in Uttar Pradesh. 
 
Methods: Total 240 samples were collected from two district of eastern plain zone of Uttar Pradesh during January 2020 to March 2021. E. coli and Klebsiella spp. isolates were confirmed by cultural characteristics in selective media and biochemical tests. ESBL producing isolates were confirmed by DDST, ESBL E-strip and PCR analysis using specific primer for (bla-CTX-M-1, bla-CTX-M-2, bla-CTX-M-9, bla-TEM and bla-SHV). Antibiogram of all confirmed ESBL producing isolates was performed against 20 antimicrobials under 12 different classes. 
 
Result: In the present study, a total of 147 (61.25%) E. coli and 19 (7.91%) Klebsiella spp. were isolated and identified based on cultural and biochemical characteristics. A total of 104 (62.65%) and 99 (59.63%) isolates were confirmed as ESBL producers using DDST and ESBL E-strip tests respectively. PCR analysis, revealed the presence of selected ESBL genes in 92 (55.42%) isolates, among them bla-CTX-M-1 was found most dominant gene (69.41%) in E. coli and (71.42%) in Klebsiella spp. All the ESBL positive E. coli and Klebsiella spp. isolates were found to be 99-100% and 86-100% sensitive to chloramphenicol, polypeptides and aminoglycosides classes respectively. Both isolates of E. coli and Klebsiella spp. showed 100% resistance to cefotaxime, cefpodoxime ceftriazone and ampicillin, 76.47% and 71.42% to ceftazidime respectively and 81.52% isolates were found to be MDR.</div>

Prevalence of Extended-spectrum &beta;-lactamase Producing E. coli and Klebsiella spp. Isolated from Buffaloes in Eastern Plain Zone 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

Transferable bla_CTX-M Carrying Multidrug Resistant Escherichia coli from Pig Population of North Eastern Region of India

Phenotypic Characterization and bla Gene Detection in Antibiotic Resistant Escherichia coli Obtained from Poultry Fecal Samples

Molecular Characterization and Antimicrobial Resistance Profiling of Extended Spectrum Beta-lactamase (ESBL) Producing Escherichia coli in Bovines from J and K, India

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

Detection and Plasmid Profiling of Extended Spectrum Beta Lactamase Producing Escherichia coli from Diarrheic Calves

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

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

Prevalence of Extended-spectrum β-lactamase Producing E. coli and Klebsiella spp. Isolated from Buffaloes in Eastern Plain Zone of Uttar Pradesh