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1.
Foodborne Pathog Dis ; 11(12): 960-5, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25405655

ABSTRACT

Between April 2009 and July 2011, 311 surface water samples in 48 cattle feedlots distributed in an area of about 67,000 km(2) were analyzed to examine the environmental dissemination of Escherichia coli O157:H7. Samples were taken inside and outside the pens, exposed and not exposed to runoff from corrals, near the feedlots. Two types of samples were defined: (1) exposed surface waters (ESW; n=251), downstream from cattle pens; and (2) nonexposed surface waters (NESW; n=60), upstream from cattle pens. By multiplex PCR, 177 (70.5%) ESW samples were rfb(O157)-positive, and 62 (24.7%) E. coli O157, and 32 (12.7%) Shiga toxin-producing E. coli (STEC) O157:H7 strains were isolated. In the NESW samples, 36 (60.0%) were rfb(O157)- positive, and 9 (15.0%) E. coli O157, and 6 (10.0%) STEC O157:H7 strains were isolated. These results showed that the environmental surface waters exposed to liquid discharges from intensive livestock operations tended to be contaminated with more STEC O157:H7 than NESW. However, no significant difference was found. This fact emphasizes the relevance of other horizontal routes of transmission, as the persistence of E. coli in the environment resulting from extensive livestock farming. By XbaI-PFGE, some patterns identified are included in the Argentine Database of E. coli O157, corresponding to strains isolated from hemolytic uremic syndrome and diarrhea cases, food, and animals, such as AREXHX01.0022, second prevalent pattern in Argentina, representing 5.5% of the total database. In the study area, characterized by the abundance of waterways, pathogens contained in feedlot runoff could reach recreational waters and also contaminate produce through irrigation, increasing the potential dissemination of STEC O157:H7 and the risk of human infections. The control of runoff systems from intensive livestock is necessary, but other alternatives should be explored to solve the problem of the presence of E. coli O157 in the aquatic rural environment.


Subject(s)
Environmental Monitoring , Escherichia coli O157/isolation & purification , Water Microbiology , Animal Husbandry , Animals , Argentina , Cattle , Electrophoresis, Gel, Pulsed-Field , Escherichia coli O157/genetics , Fresh Water/microbiology , Polymerase Chain Reaction
2.
Foodborne Pathog Dis ; 9(10): 878-84, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22994915

ABSTRACT

The purposes of this study were to detect non-O157 Shiga toxin-producing Escherichia coli (STEC) in bovine rectums and water in a beef cattle farm in Argentina, and to determine the pathogenic potential of the circulating strains. During the study, 292 rectal swabs from healthy animals and 79 environmental water samples were collected. The rectal swabs and one loop of the Moore swabs, enriched in Escherichia coli broth for 24 h at 37°C, were streaked on MacConkey agar plates and incubated overnight at 37°C. The isolates were characterized by biochemical tests and serotyped. Nonmotile STEC strains were typed for their H-specific (fliC) antigens by polymerase chain reaction (PCR). Isolates were characterized by detection of stx1, stx2, and their variants, eae, ehxA, and saa genes. Macrorestriction fragment analysis by pulsed-field gel electrophoresis (PFGE) was performed using the PulseNet standardized protocol. From 371 samples analyzed, 36.6% of rectal swabs and 34.2% of water samples were non-O157 STEC-positive by PCR, and 110 strains from rectal swabs, but only three from water, were isolated. The strains were grouped into 24 different serotypes, from which, O103:[H2] (n = 12), O136:H12 (n = 8), O178:H19 (n = 8), and O103:NM (n = 5) were most prevalent, representing 29.2% of the isolates. Predominant genotypes were stx1/eae/ehxA (16.8%) and stx2/saa/ehxA (15.9%). PFGE analysis revealed 56 different patterns, with 65 strains grouped in 19 clusters of 100% similarity. Two STEC O124:H19 strains isolated from rectal swabs and water with a 5-month interval harbored the stx1/stx2/saa/ehxA genotype, and showed an indistinguishable PFGE profile. By comparison, some XbaI-PFGE patterns identified in the present study were identical to the profiles of strains isolated from human, food, and animal sources included in the Argentine PulseNet database. By PCR, similar non-O157 detection rates were found in rectal swabs and water. However, the methodology for water samples needs to be improved, since only three strains from the total number of positive samples were recovered.


Subject(s)
Cattle/microbiology , Rectum/microbiology , Rivers/microbiology , Shiga-Toxigenic Escherichia coli/isolation & purification , Water Microbiology , Animals , Argentina , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Electrophoresis, Gel, Pulsed-Field , Feces/chemistry , Feces/microbiology , Genotype , Multigene Family , Phenotype , Polymerase Chain Reaction , Serotyping , Shiga Toxin 1/genetics , Shiga Toxin 1/isolation & purification , Shiga Toxin 2/genetics , Shiga Toxin 2/isolation & purification , Shiga-Toxigenic Escherichia coli/pathogenicity
3.
Foodborne Pathog Dis ; 7(4): 475-7, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20092405

ABSTRACT

Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen, and ruminants are recognized as the main natural reservoir. The purposes of this study were to detect E. coli O157 in bovine feces and surface water in a beef cattle farm of Gualeguaychú, Argentina; to characterize the isolates; and to establish the clonal relatedness by pulsed-field gel electrophoresis. Between September 2005 and November 2006, 288 samples of bovine feces and 79 samples of water troughs were studied. E. coli O157 was detected by immunomagnetic separation and polymerase chain reaction as screening techniques. The rfb(O157) gene was detected in 3.8% of the 288 fecal samples and in 17.7% of the 79 water samples. The stx gene was detected in all rfb(O157)-positive fecal samples and in 5.1% of water samples. Eleven E. coli O157 strains isolated from bovine fecal samples and eight from water samples were characterized. The most frequent stx genotype identified was stx(1) and stx(2c(vh-a)). Twelve (63.2%) strains harbored fliC(H7), eae, and ehxA genes. Using pulsed-field gel electrophoresis with the enzyme XbaI, a total of eight patterns with at least 72.1% similarity were identified among the 19 strains. The patterns of 15 strains were grouped into four clusters: two of them included only bovine strains and the other two only aquatic strains. No genetic correlation was established between the bovine and water STEC strains detected. The prevalence of STEC O157:H7 established in the herd studied was higher than that previously reported for Argentine grazed cattle.


Subject(s)
Cattle/microbiology , Escherichia coli O157/genetics , Escherichia coli O157/isolation & purification , Feces/microbiology , Genes, Bacterial , Rivers/microbiology , Shiga Toxin/genetics , Animal Husbandry/methods , Animals , Argentina , Bacterial Proteins/genetics , Cattle Diseases/epidemiology , Disease Reservoirs/microbiology , Disease Reservoirs/veterinary , Escherichia coli Infections/epidemiology , Escherichia coli Infections/veterinary , Escherichia coli O157/classification , Escherichia coli O157/drug effects , Microbial Sensitivity Tests/veterinary , Phylogeny , Prevalence , Seasons , Serotyping/veterinary , Shiga Toxin 1/genetics , Shiga Toxin 2/genetics , Shiga-Toxigenic Escherichia coli/classification , Shiga-Toxigenic Escherichia coli/drug effects , Shiga-Toxigenic Escherichia coli/genetics , Shiga-Toxigenic Escherichia coli/isolation & purification
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