Genes tetB Research Articles

Multidrug Resistance of Gallibacterium anatis Biovar Haemolytica Isolated from the Reproductive Tracts of Laying Hens

Antimicrobial resistance is recognized worldwide as one of the greatest threats to human and animal health and the environment. To evaluate the resistance rate of Gallibacterium anatis biovar haemolytica, which contributes to bacteremia, oophoritis, ovarian follicle degeneration, salpingitis, decreased egg production, and increased mortality in hens, strains isolated from the reproductive tracts of layers were analyzed. The oviducts were taken from three hens from each of 10 flocks manifesting clinical signs related to laying. Twenty-two isolates of G. anatis biovar haemolytica collected from the three parts of the reproductive system were identified using MALDI-TOF and molecular methods. The biovar’s resistance to 19 antimicrobial substances was assessed using the disk diffusion (n = 8) and broth microdilution (n = 11) methods. The presence of virulence (gtxA, gyrB, and flfA) and antibiotic resistance (blaROB, aphA, tetB, and tetH) genes was examined using PCR. All the isolates were resistant to four or more classes of antibiotics and were considered multidrug-resistant. All such isolates were resistant to tilmicosin, tylosin, and enrofloxacin, 88.2% were to tetracycline, and 82.4% to vancomycin. The gtxA, gyrB, tetB, and tetH genes were demonstrated. Considering the present prevalence of multidrug resistance among G. anatis biovar haemolytica isolates from laying hen reproductive tracts, surveillance in reproductive flocks is warranted.

Detection and characterization of multidrug resistant Escherichia coli carrying virulence gene isolated from broilers in Bangladesh.

The emergence and dissemination of multidrug resistant (MDR) bacteria pose a severe threat to public health by limiting clinical treatment and prophylactic options. This study investigates the prevalence of Escherichia coli in broilers, their phenotypic antimicrobial resistance (AMR) profiles and the presence of virulence-associated genes (VAGs) and antimicrobial resistance genes (ARGs) using polymerase chain reaction (PCR). A total of 216 pooled cloacal samples were collected from 1080 broilers across six districts of Bangladesh. Each pooled sample comprised randomly selected cloacal swabs from five birds per farm. E. coli isolates were identified using standard bacteriological approach, followed by biochemical assays and PCR. Antimicrobial susceptibility was assessed using the Kirby-Bauer disc diffusion method, and the presence of ARGs and VAGs was determined via PCR. Five selected isolates were partially sequenced for five VAGs using Sanger sequencing. A total of 177 E. coli isolates (81.94%, 95% confidence interval: 76.24%-86.53%) were identified. The isolates showed the highest resistance to ampicillin (93.79%), followed by tetracycline (91.53%), erythromycin (89.27%) and ciprofloxacin (87%). Conversely, ceftriaxone (80.79%) showed highest susceptibility, followed by gentamicin (37.29%) and neomycin (31.07%). All isolates were MDR, with a multiple antibiotic resistance indexes were <0.3. A significant percentage (16.38%) of E. coli isolates were MDR to five antimicrobial classes and harboured blaTEM, sul1, ere (A), tetA, tetB and tetC genes. The highest prevalent ARGs were blaTEM (88.14%) followed by ere (A) (83.62%) and sul 1 (72.32%). The prevalence of VAGs was astA (56.50%), iucD (31.07%), iss (21.47%), irp2 (15.82%) and cva/cvi (3.39%), respectively. This study highlights the presence of ARGs contributing to the development of MDR in E. coli carrying VAGs in broilers. Effective monitoring and surveillance of antimicrobial usage in poultry production systems are urgently required to prevent emergence and dissemination of AMR.

Detection of Antibiotic Resistance Genes in Escherichia coli Isolated from Chicken Eggs

Total 150 chicken eggs were randomly collected from farms, households, and markets of Rajshahi and Naogaon districts. Isolation and identification of E. coli was done by using cultural and staining characteristics, and biochemical properties. Antibiotic sensitivity assay was also performed for the isolated E. coli by using disc diffusion method. Characterization and detection of antibiotic resistance genes in E. coli isolates were done through PCR analysis. Prevalence of E. coli was 30.00% in chicken eggs in Rajshahi and Naogaon districts. Results of antibiotic sensitivity patterns of the isolated E. coli showed 57.78%, 33.33%, 31.11%, 28.89%, 26.67%, 17.78%, 15.56%, 13.33%, 13.33%, and 11.11% resistance to ciprofloxacin, doxycycline, oxytetracycline, ceftriaxone, amoxycillin, erythromycin, levofloxacin, enrofloxacin, gentamycin, and neomycin, respectively. The isolates also showed 94.44%, 82.22%, 80.00%, 77.78%, 66.67%, 60.00%, 55.56%, 48.89%, 46.67%, and 33.33% sensitive to neomycin, enrofloxacin, erythromycin, levofloxacin, gentamycin, ceftriaxone, oxytetracycline, doxycycline, amoxicillin, and ciprofloxacin, respectively. All 45 phenotypic identified isolates were confirmed as E. coli by PCR analysis. Genotypic identified isolates of E. coli were further characterized by PCR to detect antibiotic resistance genes. Out of 45 E. coli isolates 37 (82.22%) showed tetracycline resistance (tetA) gene, 32 (71.11%) showed tetracycline resistance (tetB) gene, and interestingly all 45 (100%) isolates showed quinolone group resistance (gyrA) gene by PCR analysis. J. of Sci. and Tech. Res. 5(1): 135-142, 2023

Genomic analysis of two penicillin- and rifampin-resistant Corynebacterium rouxii strains isolated from cutaneous infections in dogs

Although diphtheria is a vaccine-preventable disease, numerous cases are still reported around the world, as well as outbreaks in countries, including European ones. Species of the Corynebacterium diphtheriae complex are potentially toxigenic and, therefore, must be considered given the possible consequences, such as the circulation of clones and transmission of antimicrobial resistance and virulence genes. Recently, Corynebacterium rouxii was characterized and included among the valid species of the complex. Therefore, two cases of C. rouxii infection arising from infections in domestic animals are presented here. We provide molecular characterization, phylogenetic analyses, genome sequencing, and CRISPR-Cas analyses to contribute to a better understanding of the molecular bases, pathogenesis, and epidemiological monitoring of this species, which is still little studied. We confirmed its taxonomic position with genome sequencing and in silico analysis and identified the ST-918 for both strains. The clinical isolates were sensitive resistance to benzylpenicillin and rifampin. Antimicrobial resistance genes, including tetB, rpoB2, and rbpA genes, were predicted. The bla and ampC genes were not found. Several virulence factors were also detected, including adhesion, iron uptake systems, gene regulation (dtxR), and post-translational modification (MdbA). Finally, one prophage and the Type I-E CRISPR-Cas system were identified.

Antibiotic Resistance Profiles of Escherichia coli and Salmonella spp. Isolated From Dairy Farms and Surroundings in a Rural Area of Western Anatolia, Turkey.

Background Antibiotic resistance is a significant public health issue worldwide. Antibiotic-resistant zoonotic bacteria such as Escherichia coli (E. coli), Campylobacter, Salmonella, Listeria, Coxiella, and Mycobacterium can be particularly isolated from biofertilizers. Epidemiological studies have shown that cases of foodborne infections and intoxications are significantly related to animal-derived foods. The presence of these species in aquatic environments indicates areas or organisms contaminated with animal or human feces. Especially, the presence of E. coli in aquatic environments has become a serious problem worldwide. Pathogenic strains of E. coli cause waterborne and foodborne diseases. Materials and methods This study included a total of 290 samples collected from five different dairy farms between April and September 2023 which comprised 20 samples of cow manure, 20 samples of milk, three samples of dairy workers' hand washing water, five samples ofsoil, five samples of water, and five samples of vegetables. The samples taken from the farms were homogenized with 0.1% peptone water at a ratio of 1/10. They were then cultured on xylose lysine deoxycholate (XLD), eosin methylene blue agar (EMB), and blood agar media, and gram-negative colonies were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and the VITEK2 automated system (BioMerieux Inc., Durham, NC). Amplification of the isolated DNA extracts was performed with A.B.T.™ 2X HS-PCR MasterMix (A.B.T Laboratory Industry, Arnavutköy, Turkey) in the SimpliAmp™ thermal cycler (Thermo Fischer Scientific Inc., Waltham, MA) and visualized by agarose gel electrophoresis. Results Among the 52E. coli strains isolated in our study, the highest antibiotic sensitivity rate was observed in meropenem, while the lowest sensitivity rates were determined in cefazolin and cefuroxime. While two of the Salmonella spp. (n = 2) isolates were found to be resistant to tetracycline, and one was found to be resistant to penicillin and ampicillin. No resistance to trimethoprim/sulfamethoxazole was detected in either isolate. Extended-spectrum beta-lactamases (ESBLs) were detected in only four (7.7%) E. coli strains. While tetA, tetB, and TEM genes were seen in almost all E. coli strains, they were not found in Salmonella spp. Conclusion In conclusion, our study revealed the presence of antimicrobial resistance genes in E. coli and Salmonella spp. isolates collected from various farms and environmental samples, which render the antimicrobials used for disease treatment ineffective. Consequently, research should be undertaken to prevent the development of new resistance genes in our country, as creating new medications and treatment strategies for these diseases is costly and time-intensive.

Antimicrobial susceptibilities, resistance mechanisms and molecular characteristics of toxigenic Clostridioides difficile isolates in a large teaching hospital in Chongqing, China

ObjectivesClostridioides difficile ranks among the primary sources of healthcare-related infections and diarrhoea in numerous nations. We evaluated the drug susceptibility and resistance mechanisms of C. difficile isolates from a hospital in Chongqing, China, and identified resistance rates and resistance mechanisms that differed from previous findings. MethodsThe toxin genes and drug resistance genes of clinical strains were detected using Polymerase Chain Reaction (PCR), and these strains were subjected to Multilocus Sequence Typing (MLST). The agar dilution technique was employed for assessing susceptibility of antibiotics. Clinical data collection was completed through a review of electronic medical records. ResultsA total of 67 strains of toxin-producing C. difficile were detected. All C. difficile isolates demonstrated susceptibility to both metronidazole and vancomycin. However, resistance was observed in 8.95%, 16.42%, 56.72%, 56.72%, 31.34% and 5.97% of the isolates for tigecycline, tetracycline, clindamycin, erythromycin, moxifloxacin and rifampin, respectively. Among the strains with toxin genotypes A + B + CDT - and belonging to the ST3, six strains exhibited reduced susceptibility to tigecycline (MIC=0.5mg/L) and tetracycline (MIC=8mg/L). The tetA(P) and tetB(P) genes were present in these six strains, but were absent in tetracycline-resistant strains. Resistance genes (ermB, tetM, tetA(P) and tetB(P)) and mutations (in gyrA, gyrB, and rpoB) were identified in resistant strains. ConclusionsIn contrast to prior studies, we found higher proportions of ST3 isolates with decreased tigecycline sensitivity, sharing similar resistance patterns and resistance genes. In the resistance process of tigecycline and tetracycline, the tetA(P) and tetB(P) genes may play a weak role.

Antibiotic resistance of Gallibacterium anatis biovar haemolytica isolates from chickens.

Gallibacterium anatis is an opportunistic bacteria inducing a range of clinical signs in poultry. Gallibacterium anatis strains show multidrug resistance to antibacterial substances. The purpose of this study was to examine the susceptibility of G. anatis biovar haemolytica isolates collected from the respiratory, reproduction and gastrointestinal tracts of chickens to different antibiotics from various classes. Gallibacterium anatis biovar haemolytica was identified in tracheal swab and gastrointestinal and reproductive tract tissue samples from Polish layer and broiler chicken flocks. Twenty six isolates with β-haemolysis capability, each from a different flock, obtained from the respiratory (n = 8), reproductive (n = 10) and gastrointestinal (n = 8) tracts were selected and identified by matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry after culturing. A PCR method targeting the 16S genes was used for verification of isolates. The isolates' susceptibility to 20 antimicrobials was evaluated using the disc diffusion method for 8 drugs and the dilution method for the other 12. In addition, they were tested for the presence of the GtxA, gyrB and flfA virulence genes and blaROB, aphA, tetB and tetH antibiotic resistance genes by PCR. The most prevalent antibiotic resistance was to tilmicosin, tylosin and quinupristin/dalfopristin (all 100%), erythromycin (96.2%), tetracycline (96.2%), linezolid (92.3%) and teicoplanin (92.3%). Universal susceptibility was to only one antibiotic, chloramphenicol. Statistically significant differences were found between the resistance of gastrointestinal tract strains and that of strains from other tracts to daptomycin, gentamicin, ciprofloxacin and colistin. The GtxA and gyrB genes were detected in 100% of isolates and flfA in 19.2%. The isolates most frequently contained tetB and less frequently tetH and aphA, and did not contain blaROB. Most G. anatis biovar haemolytica isolates were resistant to many classes of antibiotics. Therefore, it is necessary and important to be vigilant for the occurrence of these bacteria and thorough in their diagnosis.

Screening of Tetracycline’s Resistance Genes among Enterobacter cloacae and Citrobacter spp. isolates

There has been a growing prevalence of tetracycline-resistant bacteria across many species and genera. As a consequence, the efficacy of tetracycline treatment has diminished with time. So this work aimed to study of tetracycline-resistant genes among Enterobacter cloacae and Citrobacter spp. Out of a total of 364 clinical specimens, 135 (37.08%) exhibited bacterial growth. Out of the total, 22 (6.04%) were classified as gram-positive bacteria, 102 (28.02%) as gram-negative bacteria, and 11 (3.02%) as mixed growth (including both gram-negative and gram-positive bacteria). The Vitek-2 system findings indicated that the proportion of Enterobacter cloacae was 25 (6.86%), whereas Citrobacter spp. accounted for just 2 (0.54%) cases. The Enterobacter cloacae and Citrobacter isolates exhibited significant levels of resistance to several antibiotics, as shown by their drug susceptibility results. The PCR findings indicated that the tetrA gene was present in 8 out of 25 isolates (32%) of E. cloacae, and in 1 out of 2 isolates (50%) of Citrobacter spp. The tetrB gene was found in 92% of E. cloacae isolates (23 out of 25) and in 100% of Citrobacter spp. isolates (2 out of 2). In this study, the tetraC gene was identified in 6 out of 25 (24%) E. cloacae isolates and in 1 out of 2 (50%) Citrobacter spp. isolates. The TetraD and TetraE genes were not identified in the current study. The amino acid sequence analysis of the tetA and tetB genes revealed a similarity rate of 83% and 95% respectively.

Multi-drug resistant (MDR) Gram-negative pathogenic bacteria isolated from poultry in the Noakhali region of Bangladesh.

Rapidly increasing antibiotic-resistant bacterial strains in Bangladesh's food and farm animals stem from the excessive and inappropriate use of antibiotics. To assess the prevalence of multi-drug resistant (MDR) Gram-negative bacteria in poultry chicks, we sought to isolate and identify strains carrying antimicrobial resistance genes. Isolation and identification involved biochemical tests, 16S rRNA sequencing, and PCR screening of species-specific genes. MDR patterns were evaluated using CLSI guidelines with seventeen antibiotics across twelve classes. Targeted gene sequences were amplified for the detection of Extended-spectrum β-Lactamase (ESBL), carbapenem, tetracycline, sulfonamide, and colistin resistance genes. Common isolates, such as Escherichia coli, Klebsiella pneumoniae, Proteus penneri, and Enterobacter hormaechei, exhibited average Multiple Antimicrobial Resistance (MAR) indices of 0.66, 0.76, 0.8, 0.84, and 0.81, 0.76, 0.84, 0.41 for broiler and layer chicken, respectively. Providencia stuartii and Salmonella enterica, exclusive to broiler samples, had MAR indices of 0.82 and 0.84, respectively. Additional isolates Morganella morganii, Aeromonas spp., and Wohlfahrtiimonas chitiniclastica were found in layers (Average MAR indices: 0.73, 0.71, and 0.91). Notably, M. morganii, E. hormaechei and W. chitiniclastica were identified for the first time in Bangladeshi poultry chicken, although their evolution is yet to be understood. In this study, Pan-drug resistance was observed in one P. stuartii (broiler) and one Aeromonas spp. (layer) with a MAR index 1, while all isolates exhibited MAR indices >0.2, indicating MDR. Antimicrobial resistance (AMR) gene screening identified blaTEM, blaSHV, tetA, and sul1 in a majority of the MDR strains. Interestingly, E. coli (lactose positive and negative) and E. hormaechei were exclusively found to possess the tetB gene. In addition, E. coli (lactose negative), Klebsiella pneumoniae, Enterobacter hormaechei, M. morganii, and P. stuartii were observed to carry the colistin-resistant mcr-1 gene, whereas sul2 was detected in E. coli (lactose positive and negative), E. hormaechei, P. stuartii, and P. penneri. These findings emphasize the health risk of our consumers of both broiler and layer chickens as they have turned into a potent reservoir of various AMR gene carrying MDR and Pan-drug resistant bacteria.

Investigation of the antimicrobial susceptibility patterns of marine cyanobacteria in Bohai Bay: Cyanobacteria may be important hosts of antibiotic resistance genes in marine environment

Marine cyanobacteria, as widely distributed and photosynthetically autotrophic bacteria in the ocean, may contribute to the global dissemination of antibiotic resistance genes (ARGs) and develop a different antimicrobial susceptibility pattern from heterotrophic bacteria and cyanobacteria from freshwater environments. However, studies on antimicrobial susceptibility and the carriage of ARGs in marine cyanobacteria are still very limited. In this study, the antibiotic resistance characteristics of cyanobacteria in nearshore waters were examined through field monitoring and laboratory investigations, which included PCR detection and ARG transformation. The results showed a positive correlation between marine cyanobacteria and some ARGs in the nearshore waters of Bohai Bay. Moreover, most screened cyanobacteria showed high minimum inhibitory concentration (MIC) values for polymyxins, tetracyclines, kanamycin, and sulfonamides, moderate MIC values for streptomycin, chloramphenicol, rifampicin, and norfloxacin, and low MIC values for roxithromycin and cephalosporins. The blaTEM, blaKPC, sul1, sul2, strA, tetA, tetB, tetC, tetM, mdfA, and intI1 genes were detected in the screened marine cyanobacteria. The highest detection rates were observed for blaTEM (93.3 %), sul1 (56.6 %), sul2 (90 %), and strA (73.3 %). The detection rate of tetA (33.3 %) was the highest among the tetracycline resistance genes, and mdfA, a multidrug-resistant pump gene with resistance to tetracycline, also showed a high detection level (23.3 %). Overall, most of the screened marine cyanobacteria were found to tolerate multiple antibiotics in seawater, and the condition of the ARGs carriage was serious. Furthermore, the screened marine Synechocystis sp. C12-2 demonstrated the ability to accept ARGs on the RP4 plasmid through natural transformation and showed reduced sensitivity to ampicillin, suggesting the possibility that some marine cyanobacteria could acquire ARGs from the environment through horizontal gene transfer. Thus, marine cyanobacteria may play an important role in the propagation of marine ARGs.

Prevalence and Antimicrobial Resistance of Escherichia coli Isolates from Chicken Meat in Romania.

The current study was conducted in order to analyze the prevalence of Escherichia coli (E. coli) in samples of chicken meat (100 chicken meat samples), as well as to evaluate the antimicrobial susceptibility of these isolates. A total of 30 samples were positive for E. coli among the collected chicken samples. Most isolates proved to be highly resistant to tetracycline (80%), ampicillin (80%), sulfamethoxazole (73.33%), chloramphenicol (70%) and nalidixic acid (60%). Strong resistance to ciprofloxacin (56.66%), trimethoprim (50%), cefotaxime (46.66%), ceftazidime (43.33%) and gentamicin (40%) was also observed. Notably, one E. coli strain also proved to be resistant to colistin. The antimicrobial resistance determinants detected among the E. coli isolates recovered in our study were consistent with their resistance phenotypes. Most of the isolates harbored the tetA (53.33%), tetB (46.66%), blaTEM (36.66%) and sul1 (26.66%) genes, but also aadA1 (23.33%), blaCTX (16.66%), blaOXA (16.66%), qnrA (16.66%) and aac (10%). In conclusion, to the best of our knowledge, this is among the first studies analyzing the prevalence and antimicrobial resistance of E. coli strains isolated from chicken meat in Romania and probably the first study reporting colistin resistance in E. coli isolates recovered from food sources in our country.

Prevalence of Escherichia coli isolated from oropharynx and trachea of clinically sick poultry and antimicrobial resistance pattern of the strains isolated

The study was carried out to investigate the isolation of Escherichia coli from tracheal and oropharyngeal swab of clinically sick chickens. The antibiotic susceptibility patterns of the isolates to several antimicrobials were determined with a striking emphasis on oxytetracycline. The PCR technique was applied to detect tetA, tetB, and tetC in the tetracycline-resistant isolates. The isolates were initially screened for their resistance patterns against 6 antimicrobials of six different groups using the disc diffusion technique. The results showed that 41% tracheal, 51% oropharyngeal, and 34% samples from both sites were E. coli positive respectively. Antimicrobial resistance profiling of the isolates revealed that all the isolates were resistant to oxytetracycline and sulphamethoxazole-trimethoprim, and also 90 %, 82.9%, 63.4%, and 39% resistant to ciprofloxacin, amoxicillin, gentamicin, and colistin respectively. Notably, 82.9% isolates (95% CI 68.4%–91.8%) showed resistance to ≥3 groups of antimicrobials that means these were multi-drug resistant. Among the tetracycline-resistant isolates, 85.4% (95% CI 71.2%–93.5%), 29.3% (7.5%–44.6%), and 7.3% (1.8% - 20.1) were positive for tetA, tetB, and tetC genes respectively. The frequency of the isolation of E. coli is greater in oropharyngeal than tracheal and both kinds of samples. Commercial poultry with E. coli strains has acquired extensive resistance to oxytetracycline. This study suggests a possible association between the tetA gene and oxytetracycline resistance in E. coli isolates, but further investigations like knockdown, whole-genome sequencing, and rescue experiments are needed to establish a direct causal relationship.

Molecular detection of genes encoding resistance to tetracycline and quinolones among Shigella strains isolated from children with acute diarrhea in southwest Iran.

An increase in the antibiotic resistance of Shigella isolates has caused major global challenges in antimicrobial therapy. Knowledge of local antibiotic resistance trends is essential for selecting appropriate antibiotic treatment regimens. This study aimed to evaluate the frequency of efflux-mediated tetracycline resistance (tet) and plasmid-mediated quinolone resistance (qnr) genes among Shigella isolates. This survey investigated 91 Shigella isolates, obtained from children with acute diarrhea. The isolates were identified using standard biochemical tests and confirmed by polymerase chain reaction (PCR) assay. Besides, the susceptibility of isolates to six selected antibiotics was assessed by the disk diffusion method. All tetracycline-resistant and nalidixic acid and ciprofloxacin resistant strains were screened for tet and qnr genes by a multiplex PCR assay. According to the results of antibiotic susceptibility tests, the highest level of antibiotic resistance was related to tetracycline (80.2%) and doxycycline (78.1%), respectively. All isolates were sensitive to tigecycline. The PCR results showed that 40.6%, 3.1%, 21.8%, 61.6% and 28.7% of the isolates carried qnrA, qnrB, qnrS, tetA, and tetB genes, respectively. None of the isolates contained tetC and tetD genes. The current findings revealed that tetA and qnrA genes might play a key role in conferring tetracycline and quinolone resistance.

Phenotypic and genotypic antimicrobial resistance correlation and plasmid characterization in Salmonella spp. isolates from Italy reveal high heterogeneity among serovars.

The spread of antimicrobial resistance among zoonotic pathogens such as Salmonella is a serious health threat, and mobile genetic elements (MGEs) carrying antimicrobial resistance genes favor this phenomenon. In this work, phenotypic antimicrobial resistance to commonly used antimicrobials was studied, and the antimicrobial resistance genes (ARGs) and plasmid replicons associated with the resistances were determined. Eighty-eight Italian Salmonella enterica strains (n = 88), from human, animal and food sources, isolated between 2009 and 2019, were selected to represent serovars with different frequency of isolation in human cases of salmonellosis. The presence of plasmid replicons was also investigated. Resistances to sulphonamides (23.9%), ciprofloxacin (27.3%), ampicillin (29.5%), and tetracycline (32.9%) were the most found phenotypes. ARGs identified in the genomes correlated with the phenotypical results, with blaTEM-1B, sul1, sul2, tetA and tetB genes being frequently identified. Point mutations in gyrA and parC genes were also detected, in addition to many different aminoglycoside-modifying genes, which, however, did not cause phenotypic resistance to aminoglycosides. Many genomes presented plasmid replicons, however, only a limited number of ARGs were predicted to be located on the contigs carrying these replicons. As an expectation of this, multiple ARGs were identified on contigs with IncQ1 plasmid replicon in strains belonging to the monophasic variant of Salmonella Typhimurium. In general, high variability in ARGs and plasmid replicons content was observed among isolates, highlighting a high level of heterogeneity in Salmonella enterica. Irrespective of the serovar., many of the ARGs, especially those associated with critically and highly important antimicrobials for human medicine were located together with plasmid replicons, thus favoring their successful dissemination.

Serotypes, Pathotypes, Shiga Toxin Variants and Antimicrobial Resistance in Diarrheagenic Escherichia coli Isolated from Rectal Swabs and Sheep Carcasses in an Abattoir in Mexico

Sheep represent one of the main reservoirs of diarrheagenic Escherichia coli; this microorganism is an etiological agent of food-borne diseases; therefore, this work aimed to identify and characterize the principal pathotypes of diarrheagenic E. coli (DEC) obtained through rectal swabs and carcasses samples from sheep slaughtered in an abattoir at the central region of Mexico. The isolates were subjected to bacteriological identification, serotyping; phylogenetic classification; detection for virulence factors, and antimicrobial sensibility. A total of 90 E. coli isolates were obtained. It was observed through 49 E. coli isolates (54%), 8 of them from carcasses, and 43 from feces was DEC. DEC serotypes with health public relevance were found: O76:H19 (n = 5), O146:H21 (n = 3), O91:H10 (n = 1), O6:NM (n = 1), and O8:NM (n = 1). Regarding the presence of Shiga toxin-producing E.coli (STEC), 43/90 (47.7%) isolates have the stx1 w/o stx2 genes, and therefore were assigned as STEC non-O157; only one isolate expressed stx1 and eae genes and was classified as t-STEC (typical STEC). Additionally, 3/90 (3.3%) harbored only the eae gene and were classified as enteropathogenic E. coli (EPEC), the stp gene was found in 2/90 isolates (2.2%) and were classified as enterotoxigenic E. coli (ETEC); 1/90 (1.1%) isolates harboring the ipaH were classified as enteroinvasive E. coli EIEC. Regarding stx1 genes subtypes, stx1c only was found in 60.5% (26/43), followed by stx1a-stx1c 20.9% (9/43) and stx1a-stx1d 2.3% (1/43). The presence of both, stx1 and stx2 genes was found in 7/43 isolates (16.3%) from rectal swabs; the combination stx1c-stx2g was detected in 3/43 isolates (6.9%), while 4 (9.4%) isolates showed different patterns (stx1a-stx1c-stx2g; stx1c-stx2b-stx2g; stx1c-stx2b and stx1a-stx1c-stx2b-stx2g). STEC isolates showed the major diversity of phylogenetic groups, although phylogroup B1 was predominant in 90.6% (39/43) while there was only one isolate (2.3%) in each remaining phylogroup (A, B2, C, and F). All EPEC, ETEC, and EIEC isolates were clustered in phylogroup B1. We observed that 27.9% (12/43) of STEC isolates carried at least one antibiotic resistance: nine isolates expressed the tetB gene, one isolate the tetA gene, two isolates the sul2 gene, one isolate the sul1 and one isolate the sul1-tetB genes. These results highlight the importance of diarrheagenic E. coli as a potential risk for public health during the slaughtering process.

Prevalence, Identification and Antibiotic Resistance of Gallibacterium anatis Isolates from Chickens in Poland.

The Gram-negative bacterium Gallibacterium anatis is part of the normal avian respiratory, intestinal and reproductive tract microflora and can be transmitted horizontally and vertically. With the coexistence of other relevant factors, G. anatis becomes an opportunistic pathogen, economically damaging to the poultry industry. This bacterium's prevalence and molecular epidemiology were investigated, and the antimicrobial treatment options for G. anatis infection in chicken flocks in Poland were assessed. Tracheal samples from 182 flocks were collected between April 2022 and March 2023. The bacterial prevalence was determined by PCR targeting the gyrB gene and 16-23S rRNA. Gallibacterium anatis was identified by matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF) after culturing and PCR amplification. Isolates' susceptibility to 11 antimicrobials was assessed with a disc diffusion test. Isolates were also tested for gyrB, GtxA and flfA virulence genes and blaROB, aphA, tetB and tetH antibiotic resistance genes by PCR. Forty-one flocks (22.5%) were positive through PCR. Antibiotic resistance was most frequently observed against tilmicosin, tylosin, enrofloxacin, amoxicillin, tetracycline and doxycycline. Multiple resistance to at least eight antibiotics occurred in 20% of isolates and to at least four in 100%. The occurrence of gyrB was noted in 100%, GtxA was detected in 89%, and flfA was found in 14% of positive samples. The tetB gene was present in 61.0% of positive samples, tetH was in 36.0%, aphA was in 16.7%, and blaROB was in 5.6%. Significant differences were found in G. anatis isolates related to the presence of the virulence genes GtxA and gyrB and the presence of resistance genes (p < 0.05) associated with resistance to tetracyclines, β-lactams and aminoglycosides. The continued rise in the resistance of G. anatis to a broadening range of antibiotics is a major problem for the poultry industry worldwide, as well as for public health. The findings of this study may expand the knowledge of the pathogenicity of G. anatis in poultry.

Antibiotic Resistance Pattern and Distribution of Resistance Genes in Salmonella Isolated from Chicken and Duck Eggs in Chhattisgarh, India

Background: Salmonella is recognized as the most prevalent bacterial cause of foodborne diseases worldwide and animal-sourced foods have been reported as a common source of Salmonella infections among humans. Methods: The commercial chicken eggs, backyard chicken eggs, and duck eggs samples, 60 each, were processed for isolation and identification of Salmonella. All Salmonella isolates were further tested for resistance against six different antibiotics. The prevalence of virulence and antimicrobial resistance genes in the Salmonella isolates was determined by PCR. Result: A total of 28 Salmonella isolates were recovered with an overall prevalence of 15.6% and out of them, 11.1% and 4.4% were from eggshell and egg content, respectively. All the isolates were found sensitive to Gentamicin however maximum resistance was observed against Cefotaxime. PCR results revealed that 100% of the isolates were carrying the invA gene however stn gene was detected in 78.6% of isolates. Among presumptively identified β-lactam-resistant Salmonella isolates, 100% and 50% isolates harbored blaTEM and blaCTX-M genes, respectively whereas none of the isolates contained the blaSHV gene. All tetracycline-resistant isolates harbored the tetA gene whereas none of the isolates carried the tetB gene. 100% of fluoroquinolone-resistant isolates were carrying the gyrA gene however parC gene was present only in 60% of isolates. These results indicate that drug-resistant Salmonella spp. were prevalent in eggs sold in the study area which can pose a serious public health problem.

Prevalence and Antimicrobial Resistance of Salmonella spp. Isolated From Chilled Chicken Meat Commercialized at Retail in Federal District, Brazil

Salmonella represents one of the most common foodborne pathogens, frequently associated with the contamination of poultry products, constituting a prominent worldwide public health concern. This study determined the prevalence and antimicrobial resistance of Salmonella spp. in chilled chicken meat (115 samples) commercialized at retail in the Federal District, Brazil. Microbiological tests were performed to screen for Salmonella spp. in the chicken meat samples, and the isolated strains were confirmed by the invA gene presence (PCR technique). The strains were evaluated for antimicrobial susceptibility by the disk diffusion technique (Kirby-Bauer method) and tested for the presence of the sul2, blaCTX, and tetB antimicrobial resistance genes. The Salmonella spp. prevalence in chilled chicken meat sold at retail in the Federal District, Brazil, was 46.1% (53 of 115 chicken meat samples analyzed had invA gene-positive strains). Seventy-eight strains of Salmonella spp. isolated from the 53 contaminated samples showed higher resistance to amoxicillin/clavulanic acid (83.3%), followed by sulfonamide (64.1%) and tetracycline (46.2%); 53.8% of the isolates were multidrug-resistant (MDR). The sul2 gene that confers resistance to sulfonamide was found in 53 strains (68.0%), the blaCTX gene that confers resistance to beta-lactams was identified in 39 strains (50.0%), and the tetB gene that confers resistance to tetracycline was identified in 29 strains (37.2%). The high percentage of Salmonella contamination in chicken meat can pose a risk to consumers’ health due to the possibility of causing salmonellosis. In addition, many isolates were MDR and carried antimicrobial resistance genes. Public agencies can use these results to develop effective public health policies and strategies to ensure the safety of these food products.

Comparison of Antimicrobial Susceptibility of Escherichia Coli Isolated from Fecal Poultry and Bovine Housed in Tunisian Farms; Phylogroup Diversity and Detection of Tetracycline and Sulfonamides Resistant Genes With Integron Class1

Little detailed documentation researched the excessive use of antimicrobials such as tetracycline and sulfonamides in veterinary medicine in Tunisia and more studies are needed. A total of 58 of commensal Escherichia coli isolates recovered from fecal samples of healthy poultry (n=31) and bovin (n=27) recovered from farms in Tunisia were examinated for 20 antimicrobial as well as, researched the presence of integron, variable regions (VRs), phylogroupes, tetracycline (tetA, tetB et tetC) and sulfonamides(sul1, sul2, sul3) resistance genes. The most frequently resistance in poultry origin were to tetracycline (94.3%), sulfonamide (70.69%), nalidixic acid (61.29), amoxicillin (58%), to trimthetoprim-sulfamethoxazole and streptomycin with the same rate (64.51%), ticarcilline (58%). Whereas, the bovine isolates were most resistant to streptomycin (55.5%), to amoxicillin (18.5), to tetracycline (37%), and have a moderate same rates to kanamycine, to trimthetoprim-sulfamethoxazole 11.11, to nalidixic acid and to sulfonamide 7.4%, For poultry and bovine class 1 integron were detected in 20, 6 isolates, respectively as well as class 2 integron were found in 2 and 1 isolates, respectively. Class 1 integrons were significantly associated with poultry origin (p=0.001). For poultry sul1, sul2, and sul3 genes were detected in 14 (46.2 %), 7 (23.8 %), and 4 (8.9 %) resistant isolates, respectively. Whereas, for bovine 5 isolates were resistant to sulfonamide and sul1 and sul2 genes were detected in 4 and 1 isolates with absence of sul3 genes. and tetracycline genes tetA, tetB genes were observed in 27 (84.37 %) and 8 (25%) resistant isolates, respectively while, TetC was not detected amongst our isolates. Seven arrangement gene cassette were detected; dfrA1-satA1-aadA1 in one identical DNA fragments with approximate size of 2000 bp and six arrangements of resistance gene cassettes of class 1 integron were detected; dfrA1+aadA1 (5isolates); for dfrA17+aadA1, dfrA12+ orfF+aadA2 each one two 2isolates; one isolate for aadA1 and dfrA5, respectively. In poultry, 16 isolates were found to belong to phylogroup A (sub- groupA1: 12, sub- groupA0: 4); 9 to B1, 1 to B2 and 5 to phylogroup D. However, in bovine 9 isolates have the phylogroups A1, 7 isolates B1, 4 isolates B2, and 3 isolates found to phylogroup D. Our results showed that the prevalence of resistance in E. coli isolates from poultry was much higher than that in bovin.

Distribution analysis of tetracycline resistance genes in Escherichia coli isolated from floor surface and effluent of pig slaughterhouses in Banten Province, Indonesia.

Slaughterhouses and their effluents could serve as a "hotspot" for the occurrence and distribution of antibiotic-resistant bacteria in the environment. This study aimed to understand the distribution of tetracycline resistance genes in Escherichia coli isolated from the floor surface and effluent samples of pig slaughterhouses in Banten Province, Indonesia. Ten samples, each from floor surface swabs and effluents, were collected from 10 pig slaughterhouses in Banten Province. Escherichia coli strains were isolated and identified by referring to the protocol of the Global Tricycle Surveillance extended-spectrum beta-lactamase E. coli from the WHO (2021). Quantitative real-time polymerase chain reaction (qPCR) was used to detect the tet genes. The tetA, tetB, tetC, tetM, tetO, and tetX genes were distributed in the isolates from the floor surface samples, and the tetA, tetC, tetE, tetM, tetO, and tetX genes were distributed in the isolates from the effluent samples. The tetO gene (60%) was the most dominant gene in the isolates from floor surface samples, while the tetA gene was the dominant one in the isolates from the effluent samples (50%). The tetA + tetO gene combination was the dominant pattern (15%) in the E. coli isolates. The high prevalence and diversity of the tet genes in floor surface and effluent samples from pig slaughterhouses in Banten Province indicated that the transmission of the tet genes had occurred from pigs to the environment; thus, this situation should be considered a serious threat to public health.