<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: Transmission of antibiotic resistance from animal food chain to human through animal food-borne pathogens have led to increased public concern. Wider surveillance on prevalence of antibiotic resistance in E. coli will provide information on evolution of resistance in various geographical locations. The purpose of this study was to investigate the presence of antimicrobial resistance of E. coli isolates from poultry under various farming system in A and N Islands and resistance genes of tet, ctx-M and aac encoding the isolates.&nbsp; 
 
Methods: Isolates were obtained from cloacal swabs in poultry under various farming systems and tested against major antimicrobial derivatives to study multi drug resistance. The presence of genes associated with resistance to tetracycline (tet A), ESBL (CTX-M) and Gentamycin (aac(3)-IV) were determined by PCR.&nbsp; 
 
Result: A total of 126 cloacal samples were analysed out of which 31.38% of the E.coli isolates from poultry under various farming systems were producing extended spectrum beta-lactamases and were multiple antimicrobial resistant. Poultry birds of commercial farms showed higher resistance levels (37.5%) than organised farms (24.76%) and desi birds (31.88%). Results indicate a high level of multi-drug resistance is emerging even in desi birds. It is suggested that an antimicrobial resistance surveillance program is needed in A and N Islands in order to detect bacterial resistance among rural poultry production as the 80 percentage of total poultry population belong to desi birds.</div>

<div>Distribution of TET, AAC and CTX-M Genes among Antibiotic Resistant Escherichia coli Isolated From Poultry under Various Farming System of A and N Islands</div>

<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

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>Background: Antibiotics are widely used in animals and human for treatment without referring an antibiogram that resulted into the proliferation of ESBL producing strains. Nowadays resistance to &beta;-lactam groups of antibiotic is expanding rapidly worldwide and threatening the public healthcare due to limited treatment options. Therefore this study aimed to detect ESBL resistance genes in E. coli and Klebsiella spp. isolated from various sources of cattle in this area of study.&nbsp; 
 
Methods: Total 240 samples were collected during August, 2019 to June, 2020, from two districts of Eastern plain zone of Uttar Pradesh. E. coli and Klebsiella spp. isolates were confirmed using uidA and 16S rRNA gene respectively. In vitro antibiotic sensitivity test was performed using disc diffusion method. ESBL producing isolates was confirmed by DDST, ESBL E-strip and PCR analysis by targeting (bla-CTX-M-1, bla-CTX-M-9, bla-TEM and bla-SHV) genes. 
 
Result: PCR analysis of these isolates confirmed 135(56.25%) as E. coli and 16(6.67%) as Klebsiella spp. Antibiotics found to be most resistant were ampicillin (89.40%) followed by cefotaxime (72.18%), cefpodoxime (71.52%), ceftriaxone (66.9%), aztreonam (61.0%) and ceftazidime (54.30%). Total 101(66.88%) isolates were confirmed as ESBL producers using DDST and 92(60.92%) by ESBL E-strip test. ESBL genes were detected in 87(57.61%) isolates by PCR analysis and among them, bla-CTX-M-1 was found most dominant gene.</div>

Detection and Molecular Characterization of Extended-spectrum &beta;-lactamase Producing E. coli and Klebsiella spp. Isolates of Cattle Origin in Eastern Plain Zone of Uttar Pradesh

<div>Background: Fracture of long bones is a common orthopaedic condition noticed in dogs and its primary goal is to completely restore the function of the injured limb as early as possible. Osteo-conductivity of hydroxyapatite can be improved further by decreasing the particle size to nanometre range and incorporation of inorganic materials in hydroxyapatite can enhance osteoblast cell material interactions. Strontium, Zinc, Silver and Fluorine are known to play an important role in the bone formation and also affect bone material characteristics such as crystallinity, degradation behaviour and mechanical properties. When doped with plasma spray nanohydroxyapatite, these multi-ions cause no harm to the physical environment during the degradation process of hydroxyapatite as these are nontoxic and play significant role in bone metabolism, growth and nourishment. Bone markers have tremendous potential as a rapid and sensitive method for assessing the response of the skeleton to medical or surgical interventions providing valuable information regarding bone turn over in animals. Hence, the current study was undertaken to evaluate the potential of multi-ion doped nano-hydroxyapatite coated intramedullary titanium implants in long bone fracture repair in dogs compared to the conventional intramedullary titanium implants through radiographical studies and evaluation of bone markers.&nbsp; 
 
Methods: Radiographical evaluation, Sandwich ELISA kits developed by Bioassay technology laboratory. 
 
Result: Plasma spray nano-hydroxyapatite coated titanium intramedullary implants have shown excellent osteo-conductivity when doped with multi-ions of Strontium, Zinc, Silver and Fluorine facilitating rapid osteoblastic activity and rapid bone turnover at the fracture site and complete fracture healing by 3rd week post-operatively as evidenced by radiographic scores and a peak BALP (Canine Bone Alkaline Phosphatase) values and early limb usage. Bone reabsorption and bone tissue remodelling due to osteoclastic action at the fracture site was quicker when the multi-ion doped nano-hydroxyapatite coated titanium intramedullary implants were used which is evidenced by the radiographic scores and highest CTX (Canine C-telopeptide of Type 1 Collagen) values indicating completion of fracture healing and near completion of bone tissue remodelling by 9th post-operative week in long bone fracture repair in dogs.</div>

​Efficacy of Multi-ion Doped Plasma Spray Nano-hydroxyapatite Coated Titanium Implants in Long Bone Fracture Repair in Dogs- Radiographical and Bone Markers Evaluation

<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: 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

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

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

Distribution of TET, AAC and CTX-M Genes among Antibiotic Resistant Escherichia coli Isolated From Poultry under Various Farming System of A and N Islands

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

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

Detection and Molecular Characterization of Extended-spectrum β-lactamase Producing E. coli and Klebsiella spp. Isolates of Cattle Origin in Eastern Plain Zone of Uttar Pradesh

Efficacy of Multi-ion Doped Plasma Spray Nano-hydroxyapatite Coated Titanium Implants in Long Bone Fracture Repair in Dogs- Radiographical and Bone Markers Evaluation

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

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

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