scholarly journals Genetic diversity and antimicrobial resistance of Escherichia coli as microbial source tracking tools of Karaj River, Iran

2017 ◽  
Vol 17 (5) ◽  
pp. 1468-1478
Author(s):  
Roohollah Kheiri ◽  
Leili Akhtari

The aim of this study was to analyze the enterobacterial repetitive intergenic consensus (ERIC)-types, phylo-groups and antimicrobial resistance (AMR) patterns of Escherichia coli and to investigate if these approaches are suitable for microbial source tracking (MST). E. coli strains were isolated from cattle faeces and Karaj River. For genetic diversity, AMR profile, and phylo-grouping, we applied ERIC-PCR, disk diffusion, and multiplex-PCR, respectively. Fifty isolates from each sample group were used in the study. ERIC fingerprinting produced ten different bands, demonstrating 64 unique and 36 repetitive profiles. Six isolates from the river showed the same ERIC pattern of the cattle, of which four expressed the same AMR profile. E. coli isolates from water were represented in A, B1, C, and D phylo-groups. Phylo-groups A, B1, and E were more prevalent in the cattle isolates and B2 was absent in both sources. Three of the water isolates with the same ERIC-type and AMR to cattle isolates showed the same phylo-groups. Genetic characteristics, AMR, and phylo-groups of the isolates from the river are diverse and complex. For accurate MST, complementary approaches should be applied together and a comprehensive library should be provided.

2005 ◽  
Vol 71 (10) ◽  
pp. 5992-5998 ◽  
Author(s):  
Zexun Lu ◽  
David Lapen ◽  
Andrew Scott ◽  
Angela Dang ◽  
Edward Topp

ABSTRACT Repetitive extragenic palindromic PCR fingerprinting of Escherichia coli is one microbial source tracking approach for identifying the host source origin of fecal pollution in aquatic systems. The construction of robust known-source libraries is expensive and requires an informed sampling strategy. In many types of farming systems, waste is stored for several months before being released into the environment. In this study we analyzed, by means of repetitive extragenic palindromic PCR using the enterobacterial repetitive intergenic consensus primers and comparative analysis using the Bionumerics software, collections of E. coli obtained from a dairy farm and from a swine farm, both of which stored their waste as a slurry in holding tanks. In all fecal samples, obtained from either barns or holding tanks, the diversity of the E. coli populations was underrepresented by collections of 500 isolates. In both the dairy and the swine farms, the diversity of the E. coli community was greater in the manure holding tank than in the barn, when they were sampled on the same date. In both farms, a comparison of stored manure samples collected several months apart suggested that the community composition changed substantially in terms of the detected number, absolute identity, and relative abundance of genotypes. Comparison of E. coli populations obtained from 10 different locations in either holding tank suggested that spatial variability in the E. coli community should be accounted for when sampling. Overall, the diversity in E. coli populations in manure slurry storage facilities is significant and likely is problematic with respect to library construction for microbial source tracking applications.


2021 ◽  
Vol 288 (1948) ◽  
Author(s):  
Katherine M. Lagerstrom ◽  
Elizabeth A. Hadly

A striking paucity of information exists on Escherichia coli in wild animals despite evidence that they harbour pathogenic and antimicrobial-resistant E. coli in their gut microbiomes and may even serve as melting pots for novel genetic combinations potentially harmful to human health. Wild animals have been implicated as the source of pathogenic E. coli outbreaks in agricultural production, but a lack of knowledge surrounding the genetics of E. coli in wild animals complicates source tracking and thus contamination curtailment efforts. As human populations continue to expand and invade wild areas, the potential for harmful microorganisms to transfer between humans and wildlife increases. Here, we conducted a literature review of the small body of work on E. coli in wild animals. We highlight the geographic and host taxonomic coverage to date, and in each, identify significant gaps. We summarize the current understanding of E. coli in wild animals, including its genetic diversity, host and geographic distribution, and transmission pathways within and between wild animal and human populations. The knowledge gaps we identify call for greater research efforts to understand the existence of E. coli in wild animals, especially in light of the potentially strong implications for global public health.


Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1013
Author(s):  
Isabel Carvalho ◽  
Rita Cunha ◽  
Carla Martins ◽  
Sandra Martínez-Álvarez ◽  
Nadia Safia Chenouf ◽  
...  

The purpose of this study was to analyse the prevalence and genetic characteristics of ESBL and acquired-AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick dogs in Portugal. Three hundred and sixty-one faecal samples from sick and healthy dogs were seeded on MacConkey agar supplemented with cefotaxime (2 µg/mL) for cefotaxime-resistant (CTXR) E. coli recovery. Antimicrobial susceptibility testing for 15 antibiotics was performed and the ESBL-phenotype of the E. coli isolates was screened. Detection of antimicrobial resistance and virulence genes, and molecular typing of the isolates (phylogroups, multilocus-sequence-typing, and specific-ST131) were performed by PCR (and sequencing when required). CTXRE. coli isolates were obtained in 51/361 faecal samples analysed (14.1%), originating from 36/234 sick dogs and 15/127 healthy dogs. Forty-seven ESBL-producing E. coli isolates were recovered from 32 sick (13.7%) and 15 healthy animals (11.8%). Different variants of blaCTX-M genes were detected among 45/47 ESBL-producers: blaCTX-M-15 (n = 26), blaCTX-M-1 (n = 10), blaCTX-M-32 (n = 3), blaCTX-M-55 (n = 3), blaCTX-M-14 (n = 2), and blaCTX-M-variant (n = 1); one ESBL-positive isolate co-produced CTX-M-15 and CMY-2 enzymes. Moreover, two additional CTXR ESBL-negative E. coli isolates were CMY-2-producers (qAmpC). Ten different sequence types were identified (ST/phylogenetic-group/β-lactamase): ST131/B2/CTX-M-15, ST617/A/CTX-M-55, ST3078/B1/CTX-M-32, ST542/A/CTX-M-14, ST57/D/CTX-M-1, ST12/B2/CTX-M-15, ST6448/B1/CTX-M-15 + CMY-2, ST5766/A/CTX-M-32, ST115/D/CMY-2 and a new-ST/D/CMY-2. Five variants of CTX-M enzymes (CTX-M-15 and CTX-M-1 predominant) and eight different clonal complexes were detected from canine ESBL-producing E. coli isolates. Although at a lower rate, CMY-2 β-lactamase was also found. Dogs remain frequent carriers of ESBL and/or qAmpC-producing E. coli with a potential zoonotic role.


2017 ◽  
Vol 15 (5) ◽  
pp. 757-765 ◽  
Author(s):  
Maria Laura Gambero ◽  
Monica Blarasin ◽  
Susana Bettera ◽  
Jesica Giuliano Albo

The genetic characteristics among Escherichia coli strains can be grouped by origin of isolation. Then, it is possible to use the genotypes as a tool to determine the source of water contamination. The aim of this study was to define water aptitude for human consumption in a rural basin and to assess the diversity of E. coli water populations. Thus, it was possible to identify the main sources of fecal contamination and to explore linkages with the hydrogeological environment and land uses. The bacteriological analysis showed that more than 50% of samples were unfit for human consumption. DNA fingerprinting analysis by BOX-PCR indicated low genotypic diversity of E. coli isolates taken from surface water and groundwater. The results suggested the presence of a dominant source of fecal contamination. The relationship between low genotypic diversity and land use would prove that water contamination comes from livestock. The genetic diversity of E. coli isolated from surface water was less than that identified in groundwater because of the different hydraulic features of both environments. Furthermore, each one of the two big strain groups identified in this basin is located in different sub-basins, showing that hydrological dynamics exerts selective pressure on bacteria DNA.


2021 ◽  
Author(s):  
Xiaoyue Li ◽  
Zewen Liu ◽  
Ningning Wang ◽  
Wei Liu ◽  
Ting Gao ◽  
...  

Abstract BackgroundIntestinal infections with Escherichia coli (E. coli) are mostly occur in piglets between 1~10 days old, which major lead to diarrhea and edema in newborn piglets. These diseases caused by E. coli can increase mortality, morbidity and growth delay of piglets, which are responsible for economic losses. In this study was to investigate the prevalence of antibiotic resistance, transmission mechanisms, and molecular epidemiology of E. coli strains isolated from pig farms in Hubei province. Furthermore, clonal and genetic diversity of isolates were identified. ResultsA total of 29 E. coli isolates were obtained from fecal of weaned piglets from Hubei province. The E. coli isolates in different regions demonstrate different genetic diversity. Multilocus sequence typing (MLST) presented that ST165 was the common sequence type, accounting for 27.6% of all E. coli isolates, followed by ST744, ST1081, ST101 and ST10. All of the isolates were resistant to the tested antibiotics to vary degrees, and more than 80% of E. coli isolates presented high resistance rates to ampicillin, lincomycin, doxycycline, tetracycline, sulfaisoxazole and ampicillin. There was one E. coli strain that was resistant to the fifteen antimicrobial agents tested. Overall, most of the isolates were conferring resistance to 5-7 antimicrobial agents tested.ConclusionsOur study reported E. coli isolates with high antimicrobial resistance and explores the genetic diversity of E. coli isolated from swine-origin. From the results obtained it can be concluded that these isolates present high prevalent multi-drug resistance. These data provide a greater understanding of the genetic diversity and antimicrobial resistance of E. coli.


2009 ◽  
Vol 55 (3) ◽  
pp. 269-276 ◽  
Author(s):  
Tanya Kon ◽  
Susan C. Weir ◽  
E. Todd Howell ◽  
Hung Lee ◽  
Jack T. Trevors

Repetitive element-polymerase chain reaction (REP-PCR) DNA fingerprinting and library-based microbial source tracking (MST) methods were utilized to investigate the potential sources of Escherichia coli pollution in recreational waters of southeastern Lake Huron. In addition to traditional sources such as humans, agriculture, and wildlife, environmentally persistent E. coli isolates were included in the identification library as a separate library unit consisting of the E. coli strains isolated from interstitial water on the beach itself. Our results demonstrated that the dominant source of E. coli pollution of the lake was agriculture, followed by environmentally adapted E. coli strains, wildlife, and then humans. A similar ratio of contributing sources was observed in all samples collected from various locations including the river discharging to the beach in both 2005 and 2006. The high similarity between the compositions of E. coli communities collected simultaneously in the river and in the lake suggests that tributaries were the major overall sources of E. coli to the lake. Our findings also suggest that environmentally adapted strains (EAS) of E. coli should be included as one of the potential sources in future microbial source tracking efforts.


2013 ◽  
Vol 79 (23) ◽  
pp. 7371-7380 ◽  
Author(s):  
Abhirosh Chandran ◽  
Asit Mazumder

ABSTRACTIn order to assess the health risk associated with a given source of fecal contamination using bacterial source tracking (BST), it is important to know the occurrence of potential pathogens as a function of host.Escherichia coliisolates (n= 593) from the feces of diverse animals were screened for various virulence genes:stx1andstx2(Shiga toxin-producingE. coli[STEC]),eaeand EAF (enteropathogenicE. coli[EPEC]), STh, STp, and LT (enterotoxigenicE. coli[ETEC]), andipaH(enteroinvasiveE. coli[EIEC]). Eleven hosts were positive for only theeae(10.11%) gene, representing atypical EPEC, while two hosts were positive for botheaeand EAF (1.3%), representing typical EPEC.stx1,stx2, or bothstx1andstx2were present in 1 (0.1%,) 10 (5.56%), and 2 (1.51%) hosts, respectively, and confirmed as non-O157 by using aE. coliO157rfb(rfbO157) TaqMan assay. STh and STp were carried by 2 hosts (2.33%) and 1 host (0.33%), respectively, while none of the hosts were positive for LT andipaH. The repetitive element palindromic PCR (rep-PCR) fingerprint analysis identified 221 unique fingerprints with a Shannon diversity index of 2.67. Multivariate analysis of variance revealed that majority of the isolates clustered according to the year of sampling. The higher prevalence of atypical EPEC and non-O157 STEC observed in different animal hosts indicates that they can be a reservoir of these pathogens with the potential to contaminate surface water and impact human health. Therefore, we suggest thatE. colifrom these sources must be included while constructing known source fingerprint libraries for tracking purposes. However, the observed genetic diversity and temporal variation need to be considered since these factors can influence the accuracy of BST results.


2010 ◽  
Vol 76 (10) ◽  
pp. 3255-3262 ◽  
Author(s):  
Linda K. Dick ◽  
Erin A. Stelzer ◽  
Erin E. Bertke ◽  
Denise L. Fong ◽  
Donald M. Stoeckel

ABSTRACT Fecal indicator bacteria (FIB), commonly used to regulate sanitary water quality, cannot discriminate among sources of contamination. The use of alternative quantitative PCR (qPCR) methods for monitoring fecal contamination or microbial source tracking requires an understanding of relationships with cultivated FIB, as contamination ages under various conditions in the environment. In this study, the decay rates of three Bacteroidales 16S rRNA gene markers (AllBac for general contamination and qHF183 and BacHum for human-associated contamination) were compared with the decay rate of cultivated Escherichia coli in river water microcosms spiked with human wastewater. The following five sets of microcosms were monitored over 11 days: control, artificial sunlight, sediment exposure, reduced temperature, and no autochthonous predation. Decay was characterized by estimation of the time needed to produce a 2-log reduction (t 99). No treatment-associated differences in the decay of the 4 targets were evident except with reduced predation, where E. coli, qHF183, and BacHum markers had lower levels of decay by day 3. However, there were substantial target-associated differences. Decay curves for the AllBac marker indicated a larger persistent population than those of the other targets. Exposure to sunlight, sediment, and reduced predation resulted in more rapid decay of the human-associated markers relative to cultivable E. coli, but there were no differences in t 99 values among the 4 targets under control conditions or at reduced temperatures. Further evaluation of epidemiological relationships will be needed in order to relate the markers directly to health risk. These findings suggest that the tested human-associated markers can complement E. coli as indicators of the human impact on sanitary water quality under the constrained conditions described in this paper.


2016 ◽  
Vol 10 (09) ◽  
pp. 920-931 ◽  
Author(s):  
Eyitayo O Adenipekun ◽  
Charlene R. Jackson ◽  
Hazem Ramadan ◽  
Bamidele A Iwalokun ◽  
Kolawole S Oyedeji ◽  
...  

Introduction: The emergence of multidrug resistance (MDR; resistance to ≥ 2 more antimicrobials) in Escherichia coli is of concern due to complications encountered in treatment. Methodology: In this study, prevalence, antimicrobial resistance, and genetic characteristics of MDR community isolates of E. coli from Lagos, Nigeria were determined. Urine and stool samples were obtained from outpatients attending Lagos State hospitals and from animal handlers in abattoirs, poultries, and open markets, from December 2012 to July 2013. Results: Approximately 50% of urine (200/394) and 88% of stool samples (120/136) were positive for E. coli. Based upon β-lactamase production, a subset of those isolates was selected for further study. Of the 22 antimicrobials tested, E. coli exhibited resistance to all antimicrobials except amikacin and piperacillin/tazobactam. The highest levels of resistance were to tetracycline (182/247; 73.7%), trimethoprim/sulfamethoxazole (152/247; 61.5%), and ampicillin (147/247; 59.1%). Resistance to the cephalosporins ranged from 1.6%–15% including the third- and fourth-generation cephalosporins, cefpodoxime (20/247; 8.1%) and cefepime (4/247; 1.6%), respectively. MDR was observed in 69.6% (172/247) of the isolates. Forty-eight E. coli resistant to at least five antimicrobials were selected for further analysis using pulsed-field gel electrophoresis; seven distinct clusters were observed among the diverse patterns. Of the 48 MDR E. coli, 30 different sequence types (ST) were detected using multilocus sequence typing, including four ST131. Conclusions: This study demonstrated circulating MDR E. coli in the Nigerian community. Monitoring of antimicrobial resistance in developing countries is necessary to optimize empiric treatment and the prudent use of antimicrobials.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pimlapas Leekitcharoenphon ◽  
Markus Hans Kristofer Johansson ◽  
Patrick Munk ◽  
Burkhard Malorny ◽  
Magdalena Skarżyńska ◽  
...  

AbstractThe emergence of antimicrobial resistance (AMR) is one of the biggest health threats globally. In addition, the use of antimicrobial drugs in humans and livestock is considered an important driver of antimicrobial resistance. The commensal microbiota, and especially the intestinal microbiota, has been shown to have an important role in the emergence of AMR. Mobile genetic elements (MGEs) also play a central role in facilitating the acquisition and spread of AMR genes. We isolated Escherichia coli (n = 627) from fecal samples in respectively 25 poultry, 28 swine, and 15 veal calf herds from 6 European countries to investigate the phylogeny of E. coli at country, animal host and farm levels. Furthermore, we examine the evolution of AMR in E. coli genomes including an association with virulence genes, plasmids and MGEs. We compared the abundance metrics retrieved from metagenomic sequencing and whole genome sequenced of E. coli isolates from the same fecal samples and farms. The E. coli isolates in this study indicated no clonality or clustering based on country of origin and genetic markers; AMR, and MGEs. Nonetheless, mobile genetic elements play a role in the acquisition of AMR and virulence genes. Additionally, an abundance of AMR was agreeable between metagenomic and whole genome sequencing analysis for several AMR classes in poultry fecal samples suggesting that metagenomics could be used as an indicator for surveillance of AMR in E. coli isolates and vice versa.


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