Background: Food borne pathogens are transmitted to humans through contaminated foods like dairy products, handmade goods and meat. Microbial biofilms are city of microbes in the form of 3D structure and attachment of bacteria with food contact surfaces lead to subsequent development of biofilms that may lead to transmission of disease and food spoilage. This study was designed to evaluate the biofilm production capability of Shiga toxin-producing Escherichia coli (STEC) isolated from milk, milk-based products and hand swabs. 
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Methods: Samples were collected and processed for isolation of E. coli. STEC were detected on the molecular basis through mPCR. Biofilm formation capability of STEC was screened by methods namely Tissue culture plate (TCP), Tube method (TM) and Congo red agar (CRA) assay.&nbsp; 
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Result: The prevalence of E. coli and STEC was found to be 22.50%, 30.55%, 12.5% and 6.25%, 4.86%, 2.08% in retail raw milk, traditional milk products and hand swabs, respectively. The overall prevalence of E. coli, serotype E. coli O157:H7 and STEC was 24.43%, 1.42% and 5.11%, respectively. The biofilm forming STEC revealed by TCP, TM and CRA were 88.88%, 77.77% and 38.88%, respectively.

<div>Evaluation of Biofilm Production Capability of Shiga Toxin Producing Escherichia coli Isolated from Milk, Milk Products and Hand Swabs&nbsp;</div>

<div style="text-align:justify">Background: Shiga toxin-producing Escherichia coli (STEC) strains are considered to be most common food-borne enteric zoonotic pathogen, causing various disease conditions in both animals and humans and are highly pathogenic to human in low infectious doses. Resistance against antibiotics by STEC is also a big concern now a days. Hence in view of the public health significance of STEC, present work was planned to know the combination of phenotypic and genotypic resistance patterns against certain most commonly used antibiotics by using disk diffusion method and multiplex PCR respectively. 
 
Methods: A total of 426 PCR confirmed STEC isolates isolated from pooled samples (animal faecal(179), farm water(122) and human faecal samples (125) of different livestock farms in and around Proddatur andhra Pradesh, were subjected to antibiotic sensitivity test by using disc diffusion method. The isolates that showed resistance for tetracycline, streptomycin, sulphonamides and ampicillin were selected and subjected to multiplex PCR for the detection of resistance genes. 
 
Result: Disk diffusion assay revealed highest phenotypic resistance for STEC isolates against Cepahlothin (100%), followed by Ampicillin (99.06%), Tetracycline (97.2%), Streptomycin (94.3%), Sulphonamides (90.8%) and Trimethoprim (84.5%). Pooled samples also revealed the presence of antimicrobial resistance genes like tetA(59.2%), tetB(43.5%), tetC (9.2%), strA (39.3%), strB(54.1%), sul1 (40.8%), sul2 (58.7%), sul3 (3.8%) and blaTEM(83.4%). These findings indicate the highest prevalence of antimicrobial resistance among the STEC isolates, which alarms indiscriminate use of antibiotics both for therapeutic purpose and as growth promoters. Strict hygienic, sanitation and HACCP programmes should be applied to counter STEC prevalence.</div>

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Detection of Antimicrobial Resistance Genes from Shiga Toxin Producing Escherichia coli by Multiplex PCR

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<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). 
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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). 
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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

<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. 
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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. 
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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. 
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Characterization of Extended Spectrum Beta Lactamase Producing Escherichia coli from Calves

<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

Background: The aim of the research is to detect Extended-Spectrum Beta-Lactamase (ESBL) producing Escherichia coli among chickens, cage swabs and wastewaters from poultry in Indonesia. 
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Methods: A total of 426 samples were collected, consisting of 200 chicken small intestine contents, 149 cage swabs and 77 wastewater. Isolation and identification of Escherichia coli were carried out on samples, followed by antimicrobial resistance testing using the Kirby-bauer diffusion test method. Phenotypic ESBL confirmation testing uses the double disc synergy test (DDST). 
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Result: Of the 264 Escherichia coli isolates, 19.3% (n = 51) were confirmed ESBL producers, out of which 58.8% (n = 30) were obtained from ayam and 41.2% (n = 21) from poultry environments. Out of a total of 21 Escherichia coli isolates from poutry environments, 80.9% (n = 17) were from cage swabs and 19.1% (n = 4) from wastewaters. Based on these results, it can be seen that chickens can be considered as a transmission medium for ESBL-producing Escherichia coli and play a role in its spread into the environment which can ultimately pose a risk to public health. Coordinated and appropriate action is needed to reduce its impact now and in the future. 
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Phenotypic Extended-spectrum Beta-lactamase Producing Escherichia coli among Chickens, Cage Swabs and Wastewaters from Poultry in Indonesia

Background: Escherichia coli infection from inside the chicken&rsquo;s body may be present if the previously slaughtered chicken was contaminated with pathogens or the cage&rsquo;s hygiene is inadequate. One of the resistance mechanisms against antibiotics used by Gram-negative bacteria of the Enterobacteriaceae family is the production of extended-spectrum &beta;-lactamases (ESBL). Research is required to determine which E. coli genotypes in broiler chicken samples collected from Surabaya&rsquo;s traditional marketplaces produce ESBL.&nbsp; 
 
Methods: A total of 100 small intestine samples collected from broiler chickens were used in this study. After initial processing all the samples were separately injected into Eosin Methylene Blue Agar (EMBA) and Indole, Methyl Red, Voges Proskauer and Citrate (IMViC) tests were used to confirm the results. A test for antibiotic sensitivity based on the Kirby-Bauer method called the Double Disk Synergy Test (DDST) was used to analyze isolates that showed resistance to aztreonam and multidrug resistance (MDR). PCR testing was performed on ESBL-verified isolates to detect the TEM and CTX-M genes. 
 
Result: The results of the test for E. coli antibiotic resistance revealed that 14 isolates were proven to be multidrug-resistant (MDR) and six of them were found to be E. coli that produced ESBL. Four isolates were found to carry the TEM and CTX-M genes after genotyping analysis. Understanding the dangers of using antibiotics as feed additives and growth promoters is crucial for farmers to address the problem of ESBL-producing E. coli bacteria in chickens. Furthermore, the government needs to take action to monitor and enforce stronger controls on the use of antibiotics, which are even readily available on the market.

Identification of the Genotypes of Extended-spectrum &beta;-lactamase Producing Escherichia coli in the Small Intestine of Broiler Chickens at a Traditional Market in Surabaya, Indonesia

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

The present study describes the prevalence of extended spectrum b-lactamases (ESBL) producing E. coli in raw chevon, milk and human samples in different districts of Chhattisgarh state. A total of 330 samples comprising of chevon (n=126), raw milk (n=104), human urine and stool (n=100) were collected from Bilaspur, Durg, Raipur, Rajnandgaon and Dhamtari districts of Chhattisgarh and processed for isolation of E. coli. The biochemically confirmed E. coli isolates were further screened of for the presence of blaTEM gene by PCR amplification. Analysis of samples indicated an overall prevalence of 31.52%. The highest prevalence of E. coli was recorded in fresh chevon samples (38.09%) followed by human urine samples (37.14%), human stool samples (30%) followed by milk samples (20.19%). In &ndash;vitro antibiotic sensitivity test of E. coli isolates revealed that all isolates to be highly sensitive towards imipenem, gentamicin, ciprofloxacin, amoxyclav, ampicillin, oxytetracyclin. The highest numbers of E. coli isolates were found resistant to erythromycin, cefotaxim, ceftazidime, cephalexin and cifixime. The 49 E. coli isolates were found to have Multiple Antibiotic Resistance (MAR) index more than 0.2, thus indicating indiscriminate use of antimicrobials. The 44 (42.3%) isolates were identified as presumptive ESBL producers and out of them 39.4% isolates were found to harbour the blaTEM gene on their plasmid DNA indicating the presence of multidrug resistant ESBL producing E. coli in foods of animal origin and human samples.

Prevalence and molecular characterization of extended-spectrum b-Lactamases (blaTEM) producing Escherichia coli isolated from humans and&nbsp;foods of animal origin in Chhattisgarh, India

<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>

Evaluation of Biofilm Production Capability of Shiga Toxin Producing Escherichia coli Isolated from Milk, Milk Products and Hand Swabs

Detection of Antimicrobial Resistance Genes from Shiga Toxin Producing Escherichia coli by Multiplex PCR

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

Characterization of Extended Spectrum Beta Lactamase Producing Escherichia coli from Calves

Detection and Characterization of β-lactamases and Biofilm-producing Escherichia coli and Klebsiella pneumoniae from Ducks and Duck Environments in West Bengal

Phenotypic Extended-spectrum Beta-lactamase Producing Escherichia coli among Chickens, Cage Swabs and Wastewaters from Poultry in Indonesia

Identification of the Genotypes of Extended-spectrum β-lactamase Producing Escherichia coli in the Small Intestine of Broiler Chickens at a Traditional Market in Surabaya, Indonesia

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

Prevalence and molecular characterization of extended-spectrum b-Lactamases (blaTEM) producing Escherichia coli isolated from humans and foods of animal origin in Chhattisgarh, India

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