Antibiotic resistance profiling and phylotyping of human-diarrheagenic Escherichia coli pathotypes detected from diarrheic and non-diarrheic calves in Iran.

BACKGROUND: Escherichia coli (E. coli) serves as a common indicator of gut microbiota and is utilized for monitoring antimicrobial resistance determinants in food-producing animals. This study aimed to investigate antimicrobial resistance patterns in virulence gene-positive E. coli isolates obtained from 340 healthy and diarrheic calves. METHODS AND RESULTS: A total of 340 fecal swab samples were obtained from diarrheic (n = 170) and healthy (n = 170) calves for 12 months from different farms in Kerman, Iran. The samples were phenotypically analyzed to detect E. coli isolates and antibiotic resistance. Also, antimicrobial resistance genes, diarrheagenic E. coli pathotypes, and phylogenetic background were screened by PCR. Fifteen percent (51/340) of E. coli isolates were positive for at least one of the examined virulence genes (VGs); the prevalence of VGs in E. coli isolates from healthy calves (36/170; 21.17%) was higher than that in diarrheic cases (15/170; 8.82%). Out of the 51 VG-positive isolates, six pathotypes including Shiga toxin-producing E. coli (STEC; 27.45%), enterotoxigenic E. coli (ETEC; 23.52%), enterohemorrhagic E. coli (EHEC; 19.6%), necrotoxigenic E. coli (NTEC; 19.6%), enteropathogenic E. coli (EPEC; 15.68%), enteroinvasive E. coli (EIEC; 1.96%) and three hybrid pathotypes including ETEC/STEC, ETEC/EHEC, and STEC/EIEC were detected among the strains. Antimicrobial resistance (AR) was observed in 98.03% of the VG-positive isolates, which was the same for both healthy and diarrheic calves. The maximum prevalence rate of AR was found against trimethoprim/sulfamethoxazole (49.01%; 3/51), while the minimum prevalence rate was against gentamycin (5.88%; 25/51). Among the VG-positives, phylotype A was found to be the most prevalent followed by B1 and D phylotypes. CONCLUSIONS: The prevalence of VG-positive E. coli isolates was higher in healthy calves compared to diarrheic cases. AR was widespread among VG-positive isolates. These findings suggest that calves may serve as potential reservoirs of antimicrobial-resistant hybrid pathotypes of E. coli.

Molecular detection of Shiga toxin-producing and antibiotic-resistant Escherichia coli isolates from buffaloes in southwest of Iran.

Three hundred fifteen bacteriological samples were obtained from feces and both external and visceral cavity surfaces of carcasses of 105 healthy buffalo slaughtered in southwest of Iran. Confirmed Escherichia coli isolates were examined for antimicrobial resistance phenotypically and were screened for stx1, stx2, and eae genes and their subtypes and assessment of antimicrobial resistance genes by regular PCR and RFLP techniques. One hundred forty-five E. coli were isolated from feces (96 isolates) and external (37) and internal (12) surfaces of carcasses. Results showed that the prevalence of STEC, EPEC, and EHEC pathotypes was 2.8%, 0.7%, and 0.7% respectively. Among 6 (4.13%) positive isolates for examined genes, 4 (2.8%) isolates were positive for stx1, 3 (2.1%) for stx2, and 2 (1.4%) for eae gene. The detected genes were classified into stx1a (4 isolates), stx2a, stx2b, stx2c, eae-ß, and unknown subtypes. The most prevalent antibiotic resistance gene was sulII (11.03%). The tetB, qnrB, floR, blaTEM, blaSHV, and aadA genes were found to a lesser extent, and all isolates were negative for blaCTX-15, blaOXA, aac(3)-I, tetA, cat1, qnrA, sulI, dhfrI, and dhfrV genes. Twelve combination patterns of antibiotic-resistant genes were observed. Maximum phenotypically resistance rate was against doxycycline (91.83%), and the minimum was against ceftazidime and florfenicol (2.75%). E. coli isolates from feces and carcasses of slaughtered buffalo can be considered a mild reservoir for stx and eae genes. However, healthy buffaloes could be considered a potential reservoir of multiple antibiotic resistance genes in E. coli isolates.

The emergence of carbapenem-resistance and New Delhi metallo-ß-lactamase-1 (blaNDM-1) among Salmonella spp. in Kerman, Iran.

Background and Objectives: Salmonella species (spp) are the most prevalent zoonotic pathogens that cause outbreaks of gastroenteritis worldwide. Therefore evaluation of the profile of antibiotic resistance, virulence factors, and plasmid replicon types in these bacteria is necessary to control and prevent the spread of potentially pathogenic and drug-resistant strains. Materials and Methods: This study was performed on 39 Salmonella spp. The antibacterial susceptibility of isolates to various antibiotic agents was determined using disk diffusion test. ß-lactamases (bla) including ESBLs, AmpC, MBLs, and virulence genes were detected by PCR methods. Plasmid incompatibility groups among the isolates were identified using PCR-based replicon typing (PBRT). Results: The most prevalent virulent gene was phoP/Q (84.6%). slyA, sopB, and stn were identified in 79.4% (n=31), 69.2% (n=27), and 2.5% (n=1) of the isolates, respectively. The antibiotic susceptibility testing showed that 30.7% of the isolates were ESBL-producing. blaTEM (41%; n=16) was the most frequent ß-lactamase gene among the isolates followed by blaNDM-1 (15.4%; n=6), blaDHA (7.7%; n=3), and blaCTX-M (1.5%; n=1). Six different plasmid replicon types, including IncP (n=9; 23%), IncFIC (n=3; 7.70%), IncY (n=3; 7.70%), IncI1-Iγ (n=2; 5.12%), IncFIIAs (n=1; 2.56%), and IncN (n=1; 2.56%) were observed among the isolates. Conclusion: Our study showed the emergence of carbapenem-resistant and blaNDM-1 among Salmonella spp. for the first time in Kerman, Iran. Since Salmonella spp. plays an important role in the transmission of resistance genes in livestock and humans in the food chains, so more stringent control policies are recommended to prevent the circulation of drug-resistant and potentially pathogenic strains from animals to humans.