Detection of numerous verotoxigenic E. coli serotypes, with multiple antibiotic resistance from cattle faeces and soil. - Related Documents




#
Rank
Similarity
Title + Abs.
Year
PMID
012345
121001.0000Detection of numerous verotoxigenic E. coli serotypes, with multiple antibiotic resistance from cattle faeces and soil. Verotoxigenic E. coli (VTEC) belong to a diverse range of serotypes. Serotypes O157 and O26 are predominately identified in VTEC-associated disease in Europe, however due to difficulty in detection little is known about the epidemiology of non-O157 serotypes. This study reports the identification of 7 VTEC serotypes from cattle faeces and soil. Cattle faeces samples (n=128) were taken from animals in 6 different farms, with soil samples (n=20) obtained from 1 farm. After sample incubation in modified tryptone soy broth (mTSB) supplemented with streptomycin sulphate samples were plated onto sorbitol MacConkey (SMAC) also supplemented with streptomycin sulphate. Bacteria detected on the plates were subjected to biochemical testing, antibiotic resistance profiling, and PCR to detect typical virulence genes, beta-lactamase and class 1 integron associated genes. Serotyping was performed on isolates positive for virulence genes. E. coli was identified from 103 samples, with verotoxin genes present in 7 E. coli isolates. Of these 7 isolates, 5 were resistant to 5 or more antibiotics. The isolate resistant to 9 antimicrobials contained a class 1 integron structure. Serotyping identified 7 separate VTEC, O2:H27, O26:H11, O63:H(-), O148:H8, O149:H1, O174:H21 and ONT:H25. Six of these VTEC have been previously associated with human disease, however with the exception of O26:H11, these serotypes have been rarely reported worldwide. Increased surveillance is required to determine the prevalence of these and other non-O157 VTEC. The presence of multi-antibiotic resistance in these isolates is of concern, and the overall implications for public health must be ascertained.200918838234
116010.9994Prevalence of shiga toxins (stx1, stx2), eaeA and hly genes of Escherichia coli O157:H7 strains among children with acute gastroenteritis in southern of Iran. OBJECTIVE: To survey the prevalence severe diarrhea arising from these bacteria in children under 5 years old in Marvdasht. METHODS: In this study faecal sample from 615 children aged <5 years old who were hospitalized for gastroenteritis in Fars hospitals in Iran were collected and then enriched in Escherichia coli (E. coli) broth and modified tryptone soy broth with novobiocin media. Fermentation of sorbitol, lactose and β -glucoronidase activity of isolated strains was examined by CT-SMAC, VRBA and chromogenic media respectively. Then isolation of E. coli O157:H7 have been confirmed with the use of specific antisera and with multiplex PCR method presence of virulence genes including: stx1, stx2, eaeA, hly has been analyzed. RESULTS: E. coli O157:H7 was detected in 7 (1.14%) stool specimens. A significant difference was seen between detection rate of isolated bacteria from age groups 18-23 months and other age groups (P=0.004). Out of considered virulence genes, only 1 of the isolated strains (0.16%) the stx1 and eaeA genes were seen and also all isolated bacteria had resistance to penicillin, ampicillin and erythromycin antibiotics. CONCLUSIONS: We found that children < 2 years of age were at highest risk of infection with E. coli O157:H7. Regarding severity of E. coli O157:H7 pathogenesis, low infectious dose and lack of routine assay for detection of these bacteria in clinical laboratory, further and completed studies on diagnosis and genotyping of this E. coli O157:H7 strain has been recommended.201525901920
267620.9993Characterization of commensal Escherichia coli isolates from slaughtered sheep in Mexico. INTRODUCTION: Commensal Escherichia coli is defined as bacteria without known virulence factors that could be playing a specific role in some diseases; however, they could be responsible to disseminate antimicrobial resistance genes to other microorganisms. This study aimed to characterize the commensal E. coli isolates obtained from slaughtered sheep in the central region of Mexico. METHODOLOGY: Isolates were classified as commensal E. coli when distinctive genes related to diarrheagenic pathotypes (stx1, stx2, eae, bfp, LT, stp, ipaH, and aggR) were discarded by PCR. Identification of serotype, phylogenetic group, and antimicrobial resistance was also performed. RESULTS: A total of 41 isolates were characterized. The phylogenetic groups found were B1 in 37 isolates (90.2%), A in 2 (4.8%), and 1 isolate (2.4%) for C and D groups. Serotypes associated with diarrhea in humans (O104:H2 and O154:NM) and hemolytic uremic syndrome (O8:NM) were detected. Thirty-three isolates (80%) were resistant to ceftazidime, 23 (56%), to tetracycline 8 (19.5%) to ampicillin, and 1 to amikacin. Six isolates (14.6%) were multidrug-resistant. CONCLUSIONS: This study provides new information about commensal E. coli in slaughtered sheep, high percentages of resistance to antibiotics, and different profiles of antimicrobial resistance were found, their dissemination constitute a risk factor towards the consuming population.202134898507
267430.9992Phylogeny, virulence factors and antimicrobial susceptibility of Escherichia coli isolated in clinical bovine mastitis. The aim of this study was to identify specific phylogeny groups, virulence genes or antimicrobial resistance traits of Escherichia coli isolated in bovine mastitis associated to clinical signs, persistence of intramammary infection in the quarter and recovery from mastitis. A total of 154 E. coli isolates from bovine clinical mastitis, 144 from the acute stage and 10 from follow-up samples 3 weeks later, originating from 144 cows in 65 dairy herds in Southern Finland were investigated. Phylogeny groups and virulence genes of the isolates were determined using polymerase chain reaction, and antimicrobial susceptibility using the VetMIC™ microdilution method. In ten cows (11.8%), infection persisted, confirmed by re-isolation of the same pulsed-field gel electrophoresis type from the affected quarter at 3 weeks post-treatment. The majority of isolates, 119 (82.6%), belonged to phylogeny group A, which mainly consisted of commensal strains. Altogether 56 isolates (38.9%) had at least one virulence gene detected. Most common virulence genes detected were irp2, iucD, papC iss; genes svg, stx1, stx2, cnf1 and hlyA were not found. Combinations of virulence genes varied greatly. Forty (27.8%) of the 144 E. coli isolates showed resistance to at least one antimicrobial tested. None of the studied phylogeny groups, virulence factors or antimicrobial resistance traits was associated with clinical signs, persistence of intramammary infection or clinical recovery from mastitis. The results support the conclusion that mastitis-causing E. coli bacteria are typical commensals.201120729012
267540.9992Prevalence and Zoonotic Risk of Multidrug-Resistant Escherichia coli in Bovine Subclinical Mastitis Milk: Insights Into the Virulence and Antimicrobial Resistance. The emergence of antibiotic-resistant microorganisms has made antimicrobial resistance a global issue, and milk is a potential source for the propagation of resistant bacteria causing zoonotic diseases. Subclinical mastitis (SCM) cases, often overlooked and mixed with normal milk in dairy farms, frequently involve E. coli, which can spread through contaminated milk. We conducted this study to determine the prevalence of virulence genes, antibiotic resistance genes (ARGs), antimicrobial susceptibility, and the genetic relatedness of multidrug-resistant (MDR) Shiga toxin-producing E. coli (STEC) isolated from SCM milk. SCM-positive bovine milk was subjected to E. coli detection using cultural, biochemical, and molecular methods. Further, we detected STEC virulence genes including stx1, stx2, and eaeA. STEC isolates were tested for ARGs including blaSHV, CITM, tetA, and aac(3)-IV, and underwent antimicrobial susceptibility tests. Moreover, we performed a phylogenetic analysis of the stx1 gene of MDR-STEC. SCM was detected in 47.2% of milk samples of which 50.54% were E. coli positive. About 17.20% of E. coli isolates contained STEC virulence genes, and stx2 was the most prevalent. Moreover, all STEC isolates harbored at least one of the ARGs, while about 43.75% of the isolates carried multiple ARGs. Additionally, all the STEC isolates showed multidrug resistance, and were found to be fully resistant against amoxicillin, followed by ampicillin (87.50%) and gentamycin (75%); and were mostly sensitive to aztreonam (81.25%) and meropenem (68.75%). In phylogeny analysis, the stx1 gene of isolated MDR-STEC showed close relatedness with disease-causing non-O157 and O157 strains of different sources including cattle, humans, and food.202539816483
115150.9992Genomic Analysis of Third Generation Cephalosporin Resistant Escherichia coli from Dairy Cow Manure. The production of extended-spectrum β-lactamases (ESBLs) conferring resistance to new derivatives of β-lactams is a major public health threat if present in pathogenic Gram-negative bacteria. The objective of this study was to characterize ceftiofur (TIO)- or cefotaxime (FOX)-resistant Escherichia coli isolated from dairy cow manure. Twenty-four manure samples were collected from four farms and incubated under anaerobic conditions for 20 weeks at 4 °C or at 25 °C. A total of 37 TIO- or FOX-resistant E. coli were isolated from two of the four farms to determine their susceptibility to 14 antibiotics. Among the 37 resistant E. coli, 10 different serotypes were identified, with O8:H1 being the predominant serotype (n = 17). Five isolates belonged to each of serotypes O9:NM and O153:H42, respectively. All 37 cephalosporin resistant isolates were multi-resistant with the most prevalent resistance spectrum being amoxicillin-clavulanic acid-ampicillin-cefoxitin-ceftiofur-ceftriaxone-chloramphenicol-streptomycin-sulfisoxazole-tetracycline-trimethoprim-sulfamethoxazole. The genomes of 18 selected isolates were then sequenced and compared to 14 selected human pathogenic E. coli reference genomes obtained from public repositories using different bioinformatics approaches. As expected, all 18 sequenced isolates carried at least one β-lactamase bla gene: TEM-1, TEM-81, CTX-M115, CTX-M15, OXA-1, or CMY-2. Several other antibiotic resistance genes (ARGs) and virulence determinants were detected in the sequenced isolates and all of them harbored antimicrobial resistance plasmids belonging to classic Inc groups. Our results confirm the presence of diverse ESBL producing E. coli isolates in dairy cow manure stored for a short period of time. Such manure might constitute a reservoir of resistance and virulence genes for other bacteria that share the same environment.201729149094
119660.9992Prediction of Phenotypic Antimicrobial Resistance Profiles From Whole Genome Sequences of Non-typhoidal Salmonella enterica. Surveillance of antimicrobial resistance (AMR) in non-typhoidal Salmonella enterica (NTS), is essential for monitoring transmission of resistance from the food chain to humans, and for establishing effective treatment protocols. We evaluated the prediction of phenotypic resistance in NTS from genotypic profiles derived from whole genome sequencing (WGS). Genes and chromosomal mutations responsible for phenotypic resistance were sought in WGS data from 3,491 NTS isolates received by Public Health England's Gastrointestinal Bacteria Reference Unit between April 2014 and March 2015. Inferred genotypic AMR profiles were compared with phenotypic susceptibilities determined for fifteen antimicrobials using EUCAST guidelines. Discrepancies between phenotypic and genotypic profiles for one or more antimicrobials were detected for 76 isolates (2.18%) although only 88/52,365 (0.17%) isolate/antimicrobial combinations were discordant. Of the discrepant results, the largest number were associated with streptomycin (67.05%, n = 59). Pan-susceptibility was observed in 2,190 isolates (62.73%). Overall, resistance to tetracyclines was most common (26.27% of isolates, n = 917) followed by sulphonamides (23.72%, n = 828) and ampicillin (21.43%, n = 748). Multidrug resistance (MDR), i.e., resistance to three or more antimicrobial classes, was detected in 848 isolates (24.29%) with resistance to ampicillin, streptomycin, sulphonamides and tetracyclines being the most common MDR profile (n = 231; 27.24%). For isolates with this profile, all but one were S. Typhimurium and 94.81% (n = 219) had the resistance determinants bla(TEM-1,)strA-strB, sul2 and tet(A). Extended-spectrum β-lactamase genes were identified in 41 isolates (1.17%) and multiple mutations in chromosomal genes associated with ciprofloxacin resistance in 82 isolates (2.35%). This study showed that WGS is suitable as a rapid means of determining AMR patterns of NTS for public health surveillance.201829636749
297270.9992Genetic characterisation of class 1 integrons among multidrug-resistant Salmonella serotypes in broiler chicken farms. OBJECTIVES: Antimicrobial resistance in Salmonella serotypes has been reported. Integrons play an important role in the dissemination of antimicrobial resistance genes in bacteria. Scarce literature is available on the identification of integrons in Salmonella isolated from broiler chickens. In this study, antimicrobial susceptibility testing and characterisation of class 1 integrons among multidrug-resistant (MDR) Salmonella enterica serotypes in broiler chicken farms in Egypt were performed. METHODS: Antimicrobial susceptibility was determined by the disk diffusion method. PCR was performed to detect antimicrobial resistance genes and class 1 integrons in the tested Salmonella serotypes. Gene sequencing of the variable region of a class 1 integron was performed. RESULTS: Salmonella spp. were detected in 26 (13.5%) of 192 broiler samples, with Salmonella Enteritidis being the most frequently detected serotype, followed by Salmonella Kentucky and Salmonella Typhimurium and other serotypes. A very high resistance rate was observed to trimethoprim/sulfamethoxazole (100%), whilst a low resistance rate was observed to cefuroxime (57.7%). MDR S. enterica isolates displayed resistance to ciprofloxacin and azithromycin. Class 1 integrons were detected in 20 (76.9%) of the 26 Salmonella isolates. A high prevalence of class 1 integrons, as the first recorded percentage in the literature, associated with MDR Salmonella isolates was observed. CONCLUSIONS: Antimicrobial resistance rates in Salmonella serotypes from broiler chicken farms were alarming, especially for ciprofloxacin and azithromycin. Thus, another therapeutic strategy other than antimicrobials is recommended to prevent outbreaks of MDR Salmonella.201829684574
119880.9992Third-Generation Cephalosporin- and Tetracycline-Resistant Escherichia coli and Antimicrobial Resistance Genes from Metagenomes of Mink Feces and Feed. American mink (Neovison vison) is a significant source of global fur production. Except for a few studies from Denmark and Canada reporting antimicrobial resistance in bacteria isolated from clinical cases, studies from the general mink population are scarce and absent in the United States. Mink feces (n = 42) and feed (n = 8) samples obtained from a mink farm were cultured for the enumeration and detection of tetracycline-resistant (TET(r))- and third-generation cephalosporin-resistant (TGC(r))-Escherichia coli. Isolates were characterized phenotypically for their resistance to other antibiotics and genotypically for resistance genes. TET(r)E. coli were detected from 98% of feces samples (mean concentration = 6 log(10)) and from 100% of feed samples (mean concentration = 3.2 logs). Among TET(r)E. coli isolates, 44% (n = 41) of fecal- and 50% (n = 8) of feed isolates were multidrug resistant (MDR; resistance to ≥3 antimicrobial classes), and 96% (n = 49) of TET(r) isolates were positive for tet(A) and/or tet(B). TGC(r)E. coli were detected from 95% of feces and 75% of feed samples with 78% (n = 40) of fecal isolates, and all six of the feed isolates were MDR. Nearly two-thirds (65%) of the TGC(r)E. coli isolates (n = 46) were positive for bla(CMY-2); the remaining 35% were positive for bla(CTX-M,) with the bla(CTX-M-14) being the predominant (75%, n = 16) variant detected. Metagenomic DNA was extracted directly from feces and feed samples, and it was tested for 84 antimicrobial resistance genes by using quantitative polymerase chain reaction (PCR) array; selected genes were also quantified by droplet digital PCR. The genes detected from the fecal samples belonged mainly to five antimicrobial classes: macrolide-lincosamide-streptogramin B (MLS(B); 100% prevalence), TETs (88.1%), β-lactams (71.4%), aminoglycosides (66.7%), and fluoroquinolones (47.6%). β-Lactam, MLS(B), and TET resistance genes were also detected from feed samples. Our study serves as a baseline for further studies and to streamline antimicrobial use in mink production in accordance with current regulations as in food animals.202133085531
131390.9992Molecular epidemiology of ceftiofur-resistant Escherichia coli isolates from dairy calves. Healthy calves (n = 96, 1 to 9 weeks old) from a dairy herd in central Pennsylvania were examined each month over a five-month period for fecal shedding of ceftiofur-resistant gram-negative bacteria. Ceftiofur-resistant Escherichia coli isolates (n = 122) were characterized by antimicrobial resistance (disk diffusion and MIC), serotype, pulsed-field gel electrophoresis subtypes, beta-lactamase genes, and virulence genes. Antibiotic disk diffusion assays showed that the isolates were resistant to ampicillin (100%), ceftiofur (100%), chloramphenicol (94%), florfenicol (93%), gentamicin (89%), spectinomycin (72%), tetracycline (98%), ticarcillin (99%), and ticarcillin-clavulanic acid (99%). All isolates were multidrug resistant and displayed elevated MICs. The E. coli isolates belonged to 42 serotypes, of which O8:H25 was the predominant serotype (49.2%). Pulsed-field gel electrophoresis classified the E. coli isolates into 27 profiles. Cluster analysis showed that 77 isolates (63.1%) belonged to one unique group. The prevalence of pathogenic E. coli was low (8%). A total of 117 ceftiofur-resistant E. coli isolates (96%) possessed the bla(CMY2) gene. Based on phenotypic and genotypic characterization, the ceftiofur-resistant E. coli isolates belonged to 59 clonal types. There was no significant relationship between calf age and clonal type. The findings of this study revealed that healthy dairy calves were rapidly colonized by antibiotic-resistant strains of E. coli shortly after birth. The high prevalence of multidrug-resistant nonpathogenic E. coli in calves could be a significant source of resistance genes to other bacteria that share the same environment.200616751500
1153100.9992Antimicrobial resistance, virulence factors and genetic diversity of Escherichia coli isolates from household water supply in Dhaka, Bangladesh. BACKGROUND: Unsafe water supplies continue to raise public health concerns, especially in urban areas in low resource countries. To understand the extent of public health risk attributed to supply water in Dhaka city, Bangladesh, Escherichia coli isolated from tap water samples collected from different locations of the city were characterized for their antibiotic resistance, pathogenic properties and genetic diversity. METHODOLOGY/PRINCIPAL FINDINGS: A total of 233 E. coli isolates obtained from 175 tap water samples were analysed for susceptibility to 16 different antibiotics and for the presence of genes associated with virulence and antibiotic resistance. Nearly 36% (n = 84) of the isolates were multi-drug(≥ 3 classes of antibiotics) resistant (MDR) and 26% (n = 22) of these were positive for extended spectrum β-lactamase (ESBL). Of the 22 ESBL-producers, 20 were positive for bla CTX-M-15, 7 for bla OXA-1-group (all had bla OXA-47) and 2 for bla CMY-2. Quinolone resistance genes, qnrS and qnrB were detected in 6 and 2 isolates, respectively. Around 7% (n = 16) of the isolates carried virulence gene(s) characteristic of pathogenic E. coli; 11 of these contained lt and/or st and thus belonged to enterotoxigenic E. coli and 5 contained bfp and eae and thus belonged to enteropathogenic E. coli. All MDR isolates carried multiple plasmids (2 to 8) of varying sizes ranging from 1.2 to >120 MDa. Ampicillin and ceftriaxone resistance were co-transferred in conjugative plasmids of 70 to 100 MDa in size, while ampicillin, trimethoprim-sulfamethoxazole and tetracycline resistance were co-transferred in conjugative plasmids of 50 to 90 MDa. Pulsed-field gel electrophoresis analysis revealed diverse genetic fingerprints of pathogenic isolates. SIGNIFICANCE: Multi-drug resistant E. coli are wide spread in public water supply in Dhaka city, Bangladesh. Transmission of resistant bacteria and plasmids through supply water pose serious threats to public health in urban areas.201323573295
2959110.9992Distribution of virulence-associated genes, antibiotic resistance and phylogenetic groups in Escherichia coli isolated from domestic and racing pigeons. Despite a lot of information about virulence and resistance of Escherichia coli (E. coli) in poultry, very limited data are currently available on its occurrence in pigeon isolates, although this poses a threat to human and animal health. Therefore, this study was conducted to explore the phylogenetic classification, antibiotic sensitivity, and virulence factors in E. coli isolated from cloacal swabs of domestic pigeons bred for meat (n = 47) and racing pigeons (n = 44). The most frequent phylogroup in racing pigeons was E (36, 82.00%), unlike domestic pigeons (B2- 19, 40.00%). The most abundant iron uptake system in both groups of bird was feoB (racing = 40, 90.90%; domestic = 44, 93.61%). The presence of ibeA (52, 57.10%) and kpsMTII (46, 50.50%) genes was detected in more than half of all strains belonging exclusively to phylogroups B2, D, E, F, clade I. Antibiotic resistance was higher in racing pigeons. All racing pigeon isolates were resistant to tetracycline and trimethoprim + sulphonamide. Resistance to ciprofloxacin was determined in three isolates (6.38%) of domestic and 33 isolates (75%) of racing pigeons. Aminoglycosides and β-lactamases resistance were also recorded. One of the important detected phenotypic mechanisms of resistance occurring in isolates from racing pigeons was AGL AAC(6´)I. Our study confirms that healthy pigeons are a reservoir of antibiotic-resistant E. coli containing an arsenal of virulence factors, thus capable of potentially causing infection. Pigeons with the option to fly to multiple places can transfer virulent and resistant bacteria. Direct contact with pigeons and their faeces and the contamination of water and food pose a threat of infection to humans and other animal species.202337076749
1143120.9992Antimicrobial Resistance and Virulence Profiles of mcr-1-Positive Escherichia coli Isolated from Swine Farms in Heilongjiang Province of China. ABSTRACT: The emergence and global distribution of the mcr-1 gene for colistin resistance have become a public concern because of threats to the role of colistin as the last line of defense against some bacteria. Because of the prevalence of mcr-1-positive Escherichia coli isolates in food animals, production of these animals has been regarded as one of the major sources of amplification and spread of mcr-1. In this study, 249 E. coli isolates were recovered from 300 fecal samples collected from swine farms in Heilongjiang Province, People's Republic of China. Susceptibility testing revealed that 186 (74.70%) of these isolates were colistin resistant, and 86 were positive for mcr-1. The mcr-1-positive isolates had extensive antimicrobial resistance profiles and additional resistance genes, including blaTEM, blaCTX-M, aac3-IV, tet(A), floR, sul1, sul2, sul3, and oqxAB. No mutations in genes pmrAB and mgrB were associated with colistin resistance. Phylogenetic group analysis revealed that the mcr-1-positive E. coli isolates belonged to groups A (52.33% of isolates), B1 (33.72%), B2 (5.81%), and D (8.14%). The prevalence of the virulence-associated genes iutA, iroN, fimH, vat, ompA, and traT was moderate. Seven mcr-1-positive isolates were identified as extraintestinal pathogenic. Among 20 mcr-1-positive E. coli isolates, multilocus sequence typing revealed that sequence type 10 was the most common (five isolates). The conjugation assays revealed that the majority of mcr-1 genes were transferable at frequencies of 7.05 × 10-7 to 7.57 × 10-4. The results of this study indicate the need for monitoring and minimizing the further dissemination of mcr-1 among E. coli isolates in food animals, particularly swine.202032730609
2138130.9992Isolation and molecular identification of multidrug-resistant Escherichia coli strains isolated from mastitic cows in Egypt. BACKGROUND: Mastitis is a common disease that affects the dairy sector globally because it not only impacts animal welfare but can also lead to significant financial losses. AIM: This study examined the phenotypic and genotypic profiles of the multidrug-resistant (MDR) Escherichia coli (E. coli) strains that were isolated from mastitic cows in Egypt to detect their pattern of antibiotic resistance. METHODS: Four hundred native breed lactating cows were evaluated to identify clinical and subclinical mastitis. A total of 100 mastitic milk samples (64 from clinical mastitis and 36 from subclinical mastitis) were collected for phenotypic isolation and identification of coliform bacteria. Escherichia coli isolates were identified through their morphological features, Gram staining, and biochemical tests. The identified E. coli strains were examined against various antibiotics using disk diffusion methods. All E. coli strains were analyzed for the antibiotic resistance genes Streptomycin (aadA), blaTEM, Tetracycline (tetA), Sulfonamides, and qnrA using PCR. RESULTS: Among 400 examined dairy cows, the prevalences of clinical and subclinical mastitis were 16% and 9%, respectively. Bacteriological isolation of coliform bacteria from mastitic milk samples revealed that E. coli was the most prevalent bacterium. Among 10 isolates of biochemically verified E. coli strains, 8 (80%) were MDR across 6 distinct classes of antibiotics. All recovered E. coli strains exhibited higher resistance to Amoxicillin, Cefotaxime, Sulphamethaxzole/Trimethoprim, and Tetracycline. High susceptibility was noticed to Ciprofloxaccin, Amoxicillin+Clavulinic, Streptomycin, Gentamicin, Chloramphenicol, and Colistin. The blaTEM gene was among the most common antibiotic resistance genes found in E. coli isolates (100%). Furthermore, the genotypes encoding resistance to tetA, aadA, and Sulfonamides were 50%, 40%, and 50%, respectively. CONCLUSION: MDR pathogenic E. coli strains are common in mastitic dairy cows in Egypt, and preventive actions must be implemented to avoid serious public health concerns.202540557079
2974140.9992Diversity of Virulence Genes in Multidrug Resistant Escherichia coli from a Hospital in Western China. BACKGROUND: Escherichia coli strains are the most commonly isolated bacteria in hospitals. The normally harmless commensal E. coli can become a highly adapted pathogen, capable of causing various diseases both in healthy and immunocompromised individuals, by acquiring a combination of mobile genetic elements. Our aim was to characterize E. coli strains from a hospital in western China to determine their virulence and antimicrobial resistance potential. METHODS: A total of 97 E. coli clinical isolates were collected from the First Affiliated Hospital of Chengdu Medical College from 2015 to 2016. Microbiological methods, PCR, and antimicrobial susceptibility tests were used in this study. RESULTS: The frequency of occurrence of the virulence genes fimC, irp2, fimH, fyuA, lpfA, hlyA, sat, and cnf1 in the E. coli isolates was 93.81, 92.78, 91.75, 84.54, 41.24, 32.99, 28.86, and 7.22%, respectively. Ninety-five (97.9%) isolates carried two or more different virulence genes. Of these, 44 (45.4%) isolates simultaneously harbored five virulence genes, 24 (24.7%) isolates harbored four virulence genes, and 17 (17.5%) isolates harbored six virulence genes. In addition, all E. coli isolates were multidrug resistant and had a high degree of antimicrobial resistance. CONCLUSION: These results indicate a high frequency of occurrence and heterogeneity of virulence gene profiles among clinical multidrug resistant E. coli isolates. Therefore, appropriate surveillance and control measures are essential to prevent the further spread of these isolates in hospitals.201931824179
2707150.9992Emergence of colistin resistance and characterization of antimicrobial resistance and virulence factors of Aeromonas hydrophila, Salmonella spp., and Vibrio cholerae isolated from hybrid red tilapia cage culture. BACKGROUND: Tilapia is a primary aquaculture fish in Thailand, but little is known about the occurrence of antimicrobial resistance (AMR) in Aeromonas hydrophila, Salmonella spp., and Vibrio cholerae colonizing healthy tilapia intended for human consumption and the co-occurrence of these AMR bacteria in the cultivation water. METHODS: This study determined the phenotype and genotype of AMR, extended-spectrum β-lactamase (ESBL) production, and virulence factors of A. hydrophila, Salmonella spp., and V. cholerae isolated from hybrid red tilapia and cultivation water in Thailand. Standard culture methods such as USFDA's BAM or ISO procedures were used for the original isolation, with all isolates confirmed by biochemical tests, serotyping, and species-specific gene detection based on PCR. RESULTS: A total of 278 isolates consisting of 15 A. hydrophila, 188 Salmonella spp., and 75 V. cholerae isolates were retrieved from a previous study. All isolates of A. hydrophila and Salmonella isolates were resistance to at least one antimicrobial, with 26.7% and 72.3% of the isolates being multidrug resistant (MDR), respectively. All A. hydrophila isolates were resistant to ampicillin (100%), followed by oxytetracycline (26.7%), tetracycline (26.7%), trimethoprim (26.7%), and oxolinic acid (20.0%). The predominant resistance genes in A. hydrophila were mcr-3 (20.0%), followed by 13.3% of isolates having floR, qnrS, sul1, sul2, and dfrA1. Salmonella isolates also exhibited a high prevalence of resistance to ampicillin (79.3%), oxolinic acid (75.5%), oxytetracycline (71.8%), chloramphenicol (62.8%), and florfenicol (55.3%). The most common resistance genes in these Salmonella isolates were qnrS (65.4%), tetA (64.9%), bla (TEM) (63.8%), and floR (55.9%). All V. cholerae isolates were susceptible to all antimicrobials tested, while the most common resistance gene was sul1 (12.0%). One isolate of A. hydrophila was positive for int1, while all isolates of Salmonella and V. cholerae isolates were negative for integrons and int (SXT). None of the bacterial isolates in this study were producing ESBL. The occurrence of mcr-3 (20.0%) in these isolates from tilapia aquaculture may signify a serious occupational and consumer health risk given that colistin is a last resort antimicrobial for treatment of Gram-negative bacteria infections. CONCLUSIONS: Findings from this study on AMR bacteria in hybrid red tilapia suggest that aquaculture as practiced in Thailand can select for ubiquitous AMR pathogens, mobile genetic elements, and an emerging reservoir of mcr and colistin-resistant bacteria. Resistant and pathogenic bacteria, such as resistance to ampicillin and tetracycline, or MDR Salmonella circulating in aquaculture, together highlight the public health concerns and foodborne risks of zoonotic pathogens in humans from cultured freshwater fish.202336855429
1147160.9992Detection and Characterisation of Colistin-Resistant Escherichia coli in Broiler Meats. The irrational use of antimicrobials has led to the emergence of resistance, impacting not only pathogenic bacteria but also commensal bacteria. Resistance against colistin, a last-resort antibiotic, mediated by globally disseminated plasmid-borne mobile colistin resistance (mcr) genes, has raised significant global concerns. This cross-sectional study aimed to investigate the antimicrobial resistance patterns of colistin-resistant Escherichia coli (E. coli) and mobilised colistin resistance (mcr 1-5) genes from broiler meat. A total of 570 broiler samples (285 liver and 285 muscle) were collected from 7 supermarkets and 11 live bird markets (LBMs) in Chattogram metropolitan areas of Bangladesh. The isolation and identification of E. coli were carried out using standard bacteriological and molecular techniques. Antimicrobial susceptibility testing (AST) was performed using the Kirby-Bauer disc diffusion method, and colistin's minimum inhibitory concentration (MIC) was determined by the broth microdilution (BMD) method. Colistin-resistant isolates were further tested for the presence of mcr (1-5) genes using polymerase chain reaction (PCR). Out of the 570 samples, 311 (54.56%; 95% confidence interval: 50.46-58.60) were positive for E. coli. AST results showed the highest resistance to sulphamethoxazole-trimethoprim (89.39%), while the highest susceptibility was observed for cefalexin (62.70%). A total of 296 isolates (95.18%) were found to be multidrug-resistant (MDR), with the multiple antibiotic resistance (MAR) index ranging from 0.38 to 1. Additionally, 41 isolates (13.18%) exhibited resistance to five antimicrobial classes, with resistance patterns of CIP + SXT + AMP + DO + TE + CT. A total of 233 isolates (74.92%) were resistant to colistin (MIC > 2 mg/L). A strong correlation between colistin resistance and the presence of the mcr-1 gene was observed (r = 1). All phenotypic colistin-resistant E. coli isolates carried the mcr-1 gene, while no isolates were positive for mcr (2-5). The detection of mcr genes in E. coli strains from poultry sources poses a significant risk, as these resistance genes can be transferred to humans through the food chain. The prevalence of multidrug-resistant Escherichia coli and the mcr-1 gene in poultry products in Bangladesh presents a significant public health and food safety concern.202439770738
1142170.9992Virulence Determinants and Plasmid-Mediated Colistin Resistance mcr Genes in Gram-Negative Bacteria Isolated From Bovine Milk. A major increase of bacterial resistance to colistin, a last-resort treatment for severe infections, was observed globally. Using colistin in livestock rearing is believed to be the ground of mobilized colistin resistance (mcr) gene circulation and is of crucial concern to public health. This study aimed to determine the frequency and virulence characteristics of colistin-resistant Gram-negative bacteria from the milk of mastitic cows and raw unpasteurized milk in Egypt. One hundred and seventeen strains belonging to Enterobacteriaceae (n = 90), Pseudomonas aeruginosa (n = 10), and Aeromonas hydrophila (n = 17) were screened for colistin resistance by antimicrobial susceptibility testing. The genetic characteristics of colistin-resistant strains were investigated for mcr-1-9 genes, phylogenetic groups, and virulence genes. Moreover, we evaluated four commonly used biocides in dairy farms for teat disinfection toward colistin-resistant strains. Multidrug-resistant (MDR) and extensive drug-resistant (XDR) phenotypes were detected in 82.91% (97/117) and 3.42% (4/117) of the isolates, respectively. Of the 117 tested isolates, 61 (52.14%) were colistin resistant (MIC >2 mg/L), distributed as 24/70 (34.29%) from clinical mastitis, 10/11 (90.91%) from subclinical mastitis, and 27/36 (75%) from raw milk. Of these 61 colistin-resistant isolates, 47 (19 from clinical mastitis, 8 from subclinical mastitis, and 20 from raw milk) harbored plasmid-borne mcr genes. The mcr-1 gene was identified in 31.91%, mcr-2 in 29.79%, mcr-3 in 34.04%, and each of mcr-4 and mcr-7 in 2.13% of the colistin-resistant isolates. Among these isolates, 42.55% (20/47) were E. coli, 21.28% (10/47) A. hydrophila, 19.12% (9/47) K. pneumoniae, and 17.02% (8/47) P. aeruginosa. This is the first report of mcr-3 and mcr-7 in P. aeruginosa. Conjugation experiments using the broth-mating technique showed successful transfer of colistin resistance to E. coli J53-recipient strain. Different combinations of virulence genes were observed among colistin-resistant isolates with almost all isolates harboring genes. Hydrogen peroxide has the best efficiency against all bacterial isolates even at a low concentration (10%). In conclusion, the dissemination of mobile colistin resistance mcr gene and its variants between MDR- and XDR-virulent Gram-negative isolates from dairy cattle confirms the spread of mcr genes at all levels; animals, humans, and environmental, and heralds the penetration of the last-resort antimicrobial against MDR bacteria. Consequently, a decision to ban colistin in food animals is urgently required to fight XDR and MDR bacteria.202134888259
2667180.9992Prevalence, virulence and antimicrobial resistance patterns of Aeromonas spp. isolated from children with diarrhea. BACKGROUND: Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. METHODS: A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. RESULTS: In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. CONCLUSION: Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially suspected to cause severe infections. Moreover, bacterial antimicrobial resistance is increasing, especially against trimethoprim-sulfamethoxazole and nalidixic acid.201627622161
1614190.9992Dissemination of Multidrug-Resistant Commensal Escherichia coli in Feedlot Lambs in Southeastern Brazil. Antimicrobial resistance (AR) is a public health issue since it limits the choices to treat infections by Escherichia coli in humans and animals. In Brazil, the ovine meat market has grown in recent years, but studies about AR in sheep are still scarce. Thus, this study aims to investigate the presence of AR in E. coli isolated from lambs during feedlot. To this end, feces from 112 lambs with 2 months of age, after weaning, were collected on the first day of the animals in the feedlot (day 0), and on the last day before slaughtering (day 42). Isolates were selected in MacConkey agar supplemented with 4 mg/L of ceftiofur and identified by biochemical methods. Isolates were submitted to an antimicrobial susceptibility test by disc-diffusion and PCR to investigate genes for phylogenetic group, virulence determinants and resistance to the several antimicrobial classes tested. The genetic localization of the bla genes detected was elucidated by S1-PFGE followed by Southern blot-hybridizations. The isolates were typed by XbaI-PFGE and MLST methods. Seventy-eight E. coli were isolated from 8/112 (7.1%) animals on day 0, and from 55/112 (49.1%) animals on day 42. Since only fimH was present in almost all E. coli (97.4%) as a virulence gene, and also 88.5% belonged to phylogroups B1 or A, we consider that isolates represent intestinal commensal bacteria. The dendrogram separated the 78 non-virulent isolates in seven clusters, two of which comprised 50 E. coli belonging to ST/CC 1727/446 or ST 3994 recovered on day 42 commonly harboring the genotype bla (CMY -2)-aac(3)-IIa -tetA-sul1-sul2-floR-cmlA. Special attention should be given to the presence of bla (CTX-M-15), a worldwide gene spread, and bla (CTX-M-14), a hitherto undetected gene in Enterobacteriaceae from food-producing animals in Brazil. Importantly, E. coli lineages and plasmids carrying bla genes detected here have already been reported as sources of infection in humans either from animals, food, or the environment, which raises public health concerns. Hence, two types of commensal E. coli carrying important AR genes clearly prevailed during feedlot, but lambs are also reservoirs of bacteria carrying important AR genes such as bla (CTX-M-14) and bla (CTX-M-15), mostly related to antimicrobial treatment failure.201931293542