<div>Background: This study was conducted to explore the contribution of poultry farms to the contamination of the environment with ESBL-producing Escherichiacoli and there with, potentially to the spread of these bacteria to humans and other animals. Hence, the present work is a poultry farm based study aimed to detect prevalence of ESBL producing E.coli among poultry of small scale farmers. 
 
Methods: ESBL-producing E. coli were detected at poultry farm (n=40). The E. coli was isolated from poultry droppings in irrespective of diseases. The required data were collected through well-structured interview schedule in farm premises. E.coli isolates were more susceptible to Gentamicin, Aztreonarm, Cefrtrazindime and Cefotoxime.&nbsp; 
 
Result: Detection of ESBL isolates was performed by Combined Disc Diffusion Methods. Out of 40 E.coli isolates 12 were phenotypically identified as ESBL producers. The prevalence of CTX-m gene is 50% and Bla (TEM) gene is 50%.&nbsp;</div>

<div>Farmer&rsquo;s Stance on Antibiotic Resistance to E. coli and Extended Spectrum - &beta; -lactamase Producing (ESBL) E. coli Isolated from Poultry Droppings</div>

Background: Rural poultry farmers are gradually changing from the backyard poultry production to intensive production, which made poultry more vulnerable to diseases and lead to indiscriminate use of antimicrobials in poultry production. As per OIE, FAO and WHO, multiple spread antimicrobial resistant pathogenic bacteria are a serious global human and animal health problem. Hence the study has been carried out to know the antimicrobial resistance status of poultry in Tirunelveli, Tamil Nadu, India. 
 
Methods: The study was aimed to find the occurrence of extended spectrum beta lactamases (ESBLs) producing E. coli from poultry origin. Caecum samples from the broiler (n=180), desi chicken (n=180) and Japanese quail (n=180) were processed for the isolation and phenotypic identification of ESBL&nbsp;E. coli. The isolates were tested for antimicrobial susceptibility test and characterization of ESBL and virulence genes done by PCR. 
 
Result: On phenotypic confirmation of ESBL producing E. coli by combined disk diffusion test (CDDT), a total of 9.63% samples were positive for extended spectrum beta lactamase producer (ESBL). Multi drug resistance (MDR) was observed in 38.23% of the broiler isolates, 27.27% and 57.14% in desi chicken and Japanese quail isolates, respectively. All the phenotypically confirmed ESBL producing E. coli isolates (n=52) were positive for uspA gene by PCR confirming the isolates as E. coli. The overall presence of the blaTEM gene was 57.69%, with broiler isolates having the highest prevalence of 67.47%, followed by desi chicken 36.36% and Japanese quail 42.85%. In contrast, the blaSHV gene was found in only 17.64% of the broiler isolates, but none were positive for the blaCTXM and blaOXA-1 gene.

Prevalence and molecular characterization of Multidrug-Resistant ESBL-Producing E. coli in commercial poultry

<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

<div>Background: Mastitis is mostly caused by mixed infections, of which S. aureus and E. coli are amongst the most important bacteria. Detection of resistant strains of these bacteria is a major public health concern due to the possibility of infection via contaminated milk and milk products intended for human consumption. This study describes simultaneous occurance of methicillin-resistant&nbsp;S. aureus and extended spectrum &beta;-lactamase producing E. coli in mastitic milk samples of buffaloes in Jabalpur, India.&nbsp; 
 
Methods: A total of 408 buffaloes milk samples were screened for mastitis by California mastitis test (CMT) test. A total of 102 mastitic milk samples were collected for isolation of Staphylococcus aureus (S. aureus) and Eschirichia coli (E. coli). Polymerase chain reaction was performed for molecular characterization of methicillin resistant S. aureus for mecA gene and ESBL producing E. coli for bla&nbsp;TEM gene, respectively. 
 
Result: On the basis of cultural and biochemical characteristics, 31/102 samples yielded&nbsp;S. aureus and 10/102 samples yielded E. coli. Out of 31 S. aureus isolates, mecA gene was positive in 05 (16.1%) and out of 10 ESBL producing E. coli isolates bla TEM was positive in 02 (20%) isolates. Coexpression of bla&nbsp;TEM and mecA genes was recorded only in 01 (10%) isolate.</div>

Coexpression of Methicillin-resistant S. aureus and ESBL Producing E. coli in Mastitic Milk of Buffaloes

Background: Salmonella spp. is a pathogenic bacterium that can cause infection in animals and human beings. Drug-resistant Salmonellae make this infection more deadly. In this background, this study was undertaken to investigate the prevalence and antimicrobial resistance pattern of ESBL / ACBL-producing Salmonella enterica strains, obtained from bovine faecal samples from different districts of West Bengal. 
&nbsp; 
Methods: A total of 170 bovine faecal samples were aseptically collected from Purba Barddhaman, Birbhum and North 24 Paraganas in this study. Standard procedures were followed to identify Salmonella enterica isolates followed by their detailed characterization and in-vitro antibiogram. 
&nbsp; 
Result: A total of 35 (20.59%) Salmonella spp. isolates were recovered after detailed morphological and biochemical characterization which were also confirmed by PCR. Higher prevalence was recorded in cross-breed jerseys (88.57%) and younger cattle within 2 yrs. of age (24.53%). A total of 18 Salmonella spp. (51.24%) isolates were detected as ESBL and 29 Salmonella spp. (82.86%) isolates were detected as ACBL producers both phenotypically. In molecular characterization, the blaAmpC (91.43%) gene was the highest followed by other genes. No TEM gene was detected in this study. Antibiogram of all the ESBL-producing Salmonella spp., isolates showed high-level resistance (58-92%) to various antibiotics like cefotaxime, doxycycline, ampicillin/cloxacillin, tetracycline, enrofloxacin etc. Antibiotics like azithromycin, chloramphenicol, HL gentamicin etc. were sensitive to these isolates. 
&nbsp;

Detection of ESBL and ACBL-producing Salmonella enterica in Cattle from Different Districts of South Bengal

<div>Background: The present study assessed the occurrence and biofilm forming potential of ESBL-producing Escherichia coli isolates recovered from raw salad vegetables collected from 32 retail vegetable shops of Mirzapur district, Uttar Pradesh, India (n=224). 
&nbsp; 
Methods: Standard bacteriological culture methods using cefotaxime-supplemented EMB agar were used for initial isolation of cefotaxime-resistant E. coli which were subjected to phenotypic detection of ESBL production by Kirby-Bauer disc diffusion test. Subsequently, all the phenotypically positive ESBL-producing E. coli isolates were subjected to determine their biofilm-producing ability by Congo red agar (CRA) assay and microtiter plate assay (MPA) followed by genotypic confirmation of biofilm production by PCR assay targeted at fim-H, Sfa and papC genes. Additionally, the isolates were genotypically explored for a potential presence of Shiga-like toxin genes (stx1 and stx2). 
&nbsp; 
Result: Altogether, 276 cefotaxime-resistant E. coli isolates were recovered, of which, 99.27% (274/276) were observed to be phenotypically positive for ESBL production. The brown, dry and rough (BDAR) and red, dry and rough (RDAR) colony morphotypes were observed by 70.80% (194/274) and 29.19% (80/274) of the isolates, respectively by CRA method revealing their potential to form biofilms. Biofilm formation was evident in 94.16% (258/274) of the isolates by MPA method, of which 79.45% (205/258), 17.05% (44/258) and 3.48% (9/258) were considered as weak, moderate and strong biofilm producers, respectively.&nbsp; The fimH gene was found to be the predominant genetic determinant of biofilm formation which was detected in 77% (211/274) of the tested isolates followed by Sfa gene (11.31%) and papC gene (4.01%). A significant difference (p&lt;0.05) was evident between CRA and MPA methods as well as phenotypic and genotypic methods for detection of biofilm formation among the recovered isolates. In addition, significant differences (p&lt;0.05) were also observed upon vegetable-wise comparison of ESBL-producing E. coli isolates and the presence of biofilm producing genes. The stx1 and stx2 genes were found to be present in one isolate recovered from a carrot sample and two isolates from coriander samples, respectively. . The findings of this study highlight the widespread occurrence of ESBL-producing E. coli from raw salad vegetables with significant biofilm-forming potential posing a public health risk. The detection of Shiga-like toxin genes in some isolates further underscores the need for stringent surveillance and improved hygiene practices to mitigate contamination risk in fresh produce.</div>

Assessment of Biofilm Formation and Shiga-like Toxin Genes in ESBL-producing E. coli Isolates Recovered from Retail Salad Vegetables in Mirzapur District, Uttar Pradesh, India

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: 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 present study was aimed to characterize ESBL producing Escherichia coli from healthy and diarrheic cattle calves. 
&nbsp; 
Methods: A total of 52 rectal swabs were collected from healthy and diarrheic cattle calves between August 2019 to March 2020. Phenotypically E. coli were characterized based on morphology, microscopy and biochemical tests. Extended Spectrum Beta Lactamases (ESBL) E. coli initial screening was performed by using cefpodoxime, ceftazidime, cefepime and aztreonam antibiotics. The confirmation of the ESBL producing strains was conducted through ESBL phenotypic identification kit and double disc diffusion test (DDDT). Molecular characterization of ESBL E. coli was conducted by PCR amplification of blaCTX-M, bla TEM and bla AmpC genes. Plasmid profiling of the ESBL E. coli isolates was carried out. 
&nbsp; 
Result: Phenotypically, out of the 51 E. coli isolated, 05 (9.8%) E. coli isolates were confirmed as ESBL strains. PCR amplification revealed 01 (20%), 04 (80%) and 03 (60%) isolates positive for blaCTX-M, bla TEM and bla AmpC genes, respectively. Plasmid profiling of the isolates showed that 60% isolates were harboring plasmids of varying size and number. The plasmid number among the isolates ranged from 0-4 plasmids per isolate and plasmid size varied from 900bp to &gt;10kb. The relatively high rates of ESBL producers in diarrheic calves were found than healthy calves. The study highlights the existence of ESBL producing E. coli isolates with certain strains carrying mobile genetic elements in livestock and probable disseminators of resistance, thus imposing public health threat. 
&nbsp;</div>

Characterization of Extended Spectrum Beta Lactamase Producing Escherichia coli from Calves

<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

<div>Background:&nbsp;Escherichia coli and Klebsiella pneumoniae are two important commensals with pathogenic potentiality in animals and birds, causing intestinal and extra-intestinal infections. These are somewhat zoonotic too via different routes. So, the present study was undertaken to investigate the prevalence of these two bacteria from cloaca and environment of ducks from different districts of West Bengal, followed by detailed characterization and antibiogram. 
 
Methods: Around 197 duck and environmental samples were collected from different districts of West Bengal. E. coli and K. pneumoniae were isolated from collected samples as per standard methods followed by serotyping of the E. coli isolates. The detection of ESBL/ACBL producing isolates was done both phenotypically and genotypically by detecting the presence of antibiotic-resistant genes. Virulence property and biofilm-producing ability of these isolates were also studied. Antibiogram of the ESBL and biofilm positive isolates were done using common 12 antibiotics by disc diffusion method.&nbsp; &nbsp; 
 
Result: A total of 134(68.02%) Escherichia coli and 33(16.75%) Klebsiella pneumoniae isolates were recovered and confirmed by PCR. The prevalence of E. coli and K. pneumoniae in cloacal swabs were less in comparison to the environmental samples (72.09% and 20.93%). Approx. 79.85% E. coli and 72.73% K. pneumoniae isolates were found to be the ESBL/ACBL producers in vitro. E. coli and K. pneumoniae isolates possessed all major antibiotic-resistant genes with the blaAmpC (64.18% &amp; 48.48%) being the most prevalent one. Twenty three (17.16%) E. coli isolates were found positive for the virulence genes, of which stx1 (39.13%) was the most common one as detected by multiplex PCR. No virulence gene was found in K. pneumoniae isolates. Ninety one (67.91%) E. coli and 26(78.79%) K. pneumoniae isolates possessed the biofilm-producing genes. All the ESBL and biofilms producing E. coli and K. pneumoniae isolates showed 100% - 95.24% resistance to ampicillin. A high percentage of resistances were also seen against cefotaxime, ticarcillin/ clavulanic acid, ertapenem, ceftazidime, norfloxacin and ciprofloxacin, which may be due to several reasons. Maximum overall sensitivity was detected against chloramphenicol, co-trimoxazole and gentamicin.</div>

Detection and Characterization of &beta;​-lactamases and Biofilm-producing Escherichia coli and Klebsiella pneumoniae from Ducks and Duck Environments in West Bengal

Farmer’s Stance on Antibiotic Resistance to E. coli and Extended Spectrum - β -lactamase Producing (ESBL) E. coli Isolated from Poultry Droppings