Dataset Information

Genetic Characterization of Antimicrobial-Resistant Escherichia coli Isolated from a Mixed-Use Watershed in Northeast Georgia, USA.

ABSTRACT: In order to determine the role of surface water in the development and spread of antibiotic-resistant (AR) bacteria, water samples were collected quarterly from 2015 to 2016 from a mixed-use watershed in Georgia. In our previous study, 496 Escherichia coli were isolated from surface water, out of which, 34 isolates were resistant to antimicrobials. For the current study, these 34 AR E. coli were characterized using pulsed-field gel electrophoresis, AR gene detection, plasmid replicon typing, class I integron detection, and multi-locus sequence typing. Genes were identified as conferring resistance to azithromycin (mph(A)); β-lactams (blaCMY, blaCTX, blaTEM); chloramphenicol (floR); streptomycin (strA, strB); sulfisoxazole (sul1, sul2); tetracycline (tetA, tetB, tetC); and trimethoprim/sulfamethoxazole (dhfr5, dhfr12). Five ciprofloxacin- and/or nalidixic-resistant isolates contained point mutations in gyrA and/or parC. Most of the isolates (n = 28) carried plasmids and three were positive for class I integrons. Twenty-nine sequence types (ST) were detected, including three epidemic urinary-tract-infection-associated ST131 isolates. One of the ST131 E. coli isolates exhibited an extended-spectrum β-lactamase (ESBL) phenotype and carried blaCTX-M-15 and blaTEM-1. To our knowledge, this is the first study on the emergence of an ESBL-producing E. coli ST131 from environmental water in the USA, which poses a potential risk to human health through the recreational, agricultural, or municipal use of this natural resource. This study identified E. coli with AR mechanisms to commonly used antimicrobials and carrying mobile genetic elements, which could transfer AR genes to other bacteria in the aquatic environment.

SUBMITTER: Cho S

PROVIDER: S-EPMC6801870 | biostudies-other | 2019 Oct

REPOSITORIES: biostudies-other

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Genetic Characterization of Antimicrobial-Resistant Escherichia coli Isolated from a Mixed-Use Watershed in Northeast Georgia, USA.

Cho Sohyun S Nguyen Hoang Anh Thi HAT McDonald Jacob M JM Woodley Tiffanie A TA Hiott Lari M LM Barrett John B JB Jackson Charlene R CR Frye Jonathan G JG

International journal of environmental research and public health 20191007 19

In order to determine the role of surface water in the development and spread of antibiotic-resistant (AR) bacteria, water samples were collected quarterly from 2015 to 2016 from a mixed-use watershed in Georgia. In our previous study, 496 Escherichia coli were isolated from surface water, out of which, 34 isolates were resistant to antimicrobials. For the current study, these 34 AR E. coli were characterized using pulsed-field gel electrophoresis, AR gene detection, plasmid replic ...[more]

PMID: 31591305

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Project description:Surface waters are important sources of water for drinking, industrial, agricultural, and recreational uses; hence, contamination of water by fecal, pathogenic, or antimicrobial resistant (AR) bacteria is a major environmental and public health concern. However, very little data is available on prevalence of these bacteria in surface water throughout a watershed. This study aimed to characterize Escherichia coli present in the Upper Oconee Watershed, a mixed-use watershed in Athens, GA, USA for potential pathogenicity and AR. E. coli were enumerated by colony counts, cultured by enrichment and direct plating, and characterized by phylo-groups, diarrheagenic pathotypes, and antimicrobial susceptibility. From the analysis, 99.3% (455/458) of the total samples were positive for E. coli resulting in 496 isolates. E. coli counts were as high as 1.2×104 CFU/100 ml, which is above the United States Environmental Protection Agency (U.S. EPA) threshold for recreational water (235 CFU/100 ml based on a one-time measurement). Phylo-groups B2 (31.7%; 157/496) and B1 (30.8%; 153/496) were the most prevalent among the isolates. Enteropathogenic E. coli (EPEC) (19/496) and Shiga toxin-producing E. coli (STEC) (1/496) were the only diarrheagenic pathotypes detected. AR was observed in 6.9% (34/496) of the isolates, 15 of which were multidrug resistant (MDR; resistance to two or more classes of antimicrobials). Tetracycline resistance was most often detected (76.5%; 26/34), followed by ampicillin (32.4%; 11/34), streptomycin (23.5%; 8/34), sulfisoxazole (23.5%; 8/34), and nalidixic acid (14.7%; 5/34). Results from this study showed that E. coli is prevalent in high levels in the Upper Oconee Watershed, suggesting possible widespread fecal contamination. The presence of pathogenic, AR E. coli in the watershed indicates that environmental water can serve as a reservoir of resistant bacteria that may be transferred to humans through drinking and recreational activities.

Project description:To characterize the differentially expressed genes between pathogenic avian E. coli and human E. coli ATCC 25922, Abstract Escherichia coli (E. coli) is a harmless common bacterium of poultry intestine, but with a wide range of genomic flexibility, is also causative agent of many poultry diseases collectively called colibacillosis that is blamed for high economic loss in poultry sector worldwide. Numerous studies have been conducted to check the prevalence of pathogenic E. coli in poultry and poultry products, however limited data are available regarding their resistance and virulence associated genes expression profile. This study examined the pathogenomic content of poultry E. coli by antibiotic susceptibility, biofilm formation and adhesion, invasion and intracellular survivability assays in Caco-2 and Raw 264.7 cell lines along with the determination of median lethal dose in two-day old chickens. A clinical pathogenic multidrug resistant (MDR) isolate, E. coli 381, isolated from broilers was found to be highly virulent in cell culture and in chicken model. Transcriptome analysis has been skewed towards bacterial pathogens because of the prioritization of poultry diseases. Comparative gene expression profile of MDR E. coli 381 and the reference human strain E. coli ATCC 25922 was done using Illumina HiSeq2500 transcriptome and results were verified by RT-qPCR analyses. A number of resistant encoding genes including multidrug transporters, multidrug resistance proteins, porins and autotransporters were identified. We also noticed overexpression of very important virulent genes (fimA, fimC, fimH and fimI) encoding the type-1 fimbrial proteins, curli fimbriae genes , invasin genes, toxin-encoding genes and biofilm forming regulatory genes . In addition, many types of stress and metal homeostasis controlling genes were among up-regulated genes in E. coli 381 as compared to reference strain. GO and KEGG pathway analysis results revealed that genes controlling secondary metabolism, drug transport, adhesion and invasion proteins, and mobile genetic elements were over-expressed in E. coli 381. Several genes involved in cellular and metabolic processes such as carbohydrate metabolism were responsible for stress tolerance. Seminal description of the transcriptomic results and other unique features of E. coli 381 confirmed that it is highly virulent and MDR strain of poultry origin. This comparative study provides new avenues for further work on molecular mechanisms to prevent resistance development in bacteria and to ensure public health.

Project description:Decontamination practices, which often involve thermal treatments, are routinely performed in beef packing plants and have generally improved the safety of meat in North America. We investigated whether Escherichia coli in the beef production chain is becoming more heat resistant due to those treatments. Cattle isolates (n?=?750) included seven serogroups (O157, O103, O111, O121, O145, O26, and O45) which were collected between 2002 and 2017. Beef plant isolates (n?=?700) from carcasses, fabrication equipment, and beef products were included. Heat resistance was determined in Luria-Bertani broth at 60°C and by PCR screening for the locus of heat resistance (LHR). The decimal reduction for E. coli at 60°C (D 60ºC values) ranged from 0 to 7.54?min, with 97.2% of the values being?<2?min. The prevalence of E. coli with D 60ºC values?of >2?min was not significantly different (P?>?0.05) among cattle and meat plant isolates. E. coli from equipment before sanitation (median, 1.03?min) was more heat resistant than that after sanitation (median, 0.9?min). No significant difference in D 60ºC values was observed among E. coli isolates from different years, from carcasses before and after antimicrobial interventions, or from before and during carcass chilling. Of all isolates, 1.97% harbored LHR, and the LHR-positive isolates had greater median D 60ºC values than the LHR-negative isolates (3.25 versus 0.96?min). No increase in heat resistance in E. coli was observed along the beef production chain or with time.IMPORTANCE The implementation of multiple hurdles in the beef production chain has resulted in substantial improvement in the microbial safety of beef in Canada. In this study, we characterized a large number of Escherichia coli isolates (n?=?1,450) from various sources/stages of beef processing to determine whether the commonly used antimicrobial interventions would give rise to heat-resistant E. coli on meat, which in turn may require alternatives to the current control of pathogens and/or modifications to the current cooking recommendations for meat. The findings show that the degree and rate of heat resistance in E. coli did not increase along the production chain or with time. This furthers our understanding of man-made ecological niches that are required for the development of heat resistance in E. coli.

Project description:There is growing evidence that anthropogenic sources of antibiotics and antimicrobial-resistant bacteria can spill over into natural ecosystems, raising questions about the role wild animals play in the emergence, maintenance, and dispersal of antibiotic resistance genes. In particular, we lack an understanding of how resistance genes circulate within wild animal populations, including whether specific host characteristics, such as social associations, promote interhost transmission of these genes. In this study, we used social network analysis to explore the forces shaping population-level patterns of resistant Escherichia coli in wild giraffe (Giraffa camelopardalis) and assess the relative importance of social contact for the dissemination of resistant E. coli between giraffe. Of 195 giraffe sampled, only 5.1% harbored E. coli isolates resistant to one or more tested antibiotics. Whole-genome sequencing on a subset of resistant isolates revealed a number of acquired resistance genes with linkages to mobile genetic elements. However, we found no evidence that the spread of resistance genes among giraffe was facilitated by interhost associations. Giraffe with lower social degree were more likely to harbor resistant E. coli, but this relationship was likely driven by a correlation between an individual's social connectedness and age. Indeed, resistant E. coli was most frequently detected in socially isolated neonates, indicating that resistant E. coli may have a selective advantage in the gastrointestinal tracts of neonates compared to other age classes. Taken together, these results suggest that the maintenance of antimicrobial-resistant bacteria in wild populations may, in part, be determined by host traits and microbial competition dynamics within the host.IMPORTANCE Antimicrobial resistance represents a significant threat to human health, food security, and the global economy. To fully understand the evolution and dissemination of resistance genes, a complete picture of antimicrobial resistance in all biological compartments, including natural ecosystems, is required. The environment and wild animals may act as reservoirs for anthropogenically derived resistance genes that could be transferrable to clinically relevant bacteria of humans and domestic animals. Our study investigated the possible transmission mechanisms for antimicrobial-resistant bacteria within a wild animal population and, more broadly, contributes to our understanding of how resistance genes are spread and maintained in natural ecosystems.

Dataset Information

Genetic Characterization of Antimicrobial-Resistant Escherichia coli Isolated from a Mixed-Use Watershed in Northeast Georgia, USA.

Publications

Genetic Characterization of Antimicrobial-Resistant <i>Escherichia coli</i> Isolated from a Mixed-Use Watershed in Northeast Georgia, USA.

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