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1.
Rev Argent Microbiol ; 2024 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-39277447

RESUMO

Rectal swabs (122) from pediatric patients were analyzed by polymerase chain reaction (PCR) for the detection of EPEC and STEC. STEC isolates were tested for the presence of stx1, stx2, eae, saa and ehxA. All eae-positive samples were tested for the presence of bfpA, and antigen O was determined using the agglutination test. Int1 and Int2 were detected to identify the presence of integrons class 1 and 2, respectively. Escherichia coli was detected in 68% of the samples, of which 18.8% were STEC (2.45%) and EPEC (16.3%). Serogroups STEC O145 and EPEC O130, O113 and O157 were observed, while three strains were non-typable. None of the EPEC strains carrying tbfpA and class 1 and 2 integrons was detected in any of the samples. The results obtained are important considering the virulence profiles found in the isolated EPEC and STEC strains and the serogroups associated with disease in humans.

2.
Int J Mol Sci ; 24(3)2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36769094

RESUMO

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen that causes gastroenteritis and Hemolytic Uremic Syndrome. Cattle are the main animal reservoir, excreting the bacteria in their feces and contaminating the environment. In addition, meat can be contaminated by releasing the intestinal content during slaughtering. Here, we evaluated the safety and immunogenicity of a vaccine candidate against STEC that was formulated with two chimeric proteins (Chi1 and Chi2), which contain epitopes of the OmpT, Cah and Hes proteins. Thirty pregnant cows in their third trimester of gestation were included and distributed into six groups (n = 5 per group): four groups were administered intramuscularly with three doses of the formulation containing 40 µg or 100 µg of each protein plus the Quil-A or Montanide™ Gel adjuvants, while two control groups were administered with placebos. No local or systemic adverse effects were observed during the study, and hematological parameters and values of blood biochemical indicators were similar among all groups. Furthermore, all vaccine formulations triggered systemic anti-Chi1/Chi2 IgG antibody levels that were significantly higher than the control groups. However, specific IgA levels were generally low and without significant differences among groups. Notably, anti-Chi1/Chi2 IgG antibody levels in the serum of newborn calves fed with colostrum from their immunized dams were significantly higher compared to newborn calves fed with colostrum from control cows, suggesting a passive immunization through colostrum. These results demonstrate that this vaccine is safe and immunogenic when applied to pregnant cows during the third trimester of gestation.


Assuntos
Infecções por Escherichia coli , Síndrome Hemolítico-Urêmica , Escherichia coli Shiga Toxigênica , Vacinas de Subunidades Antigênicas , Animais , Bovinos , Feminino , Gravidez , Infecções por Escherichia coli/prevenção & controle , Infecções por Escherichia coli/veterinária , Imunização Passiva , Imunoglobulina G , Vacinas de Subunidades Antigênicas/efeitos adversos
3.
Rev Argent Microbiol ; 52(2): 131-135, 2020.
Artigo em Espanhol | MEDLINE | ID: mdl-31635897

RESUMO

Shiga toxin-producing Escherichia coli (STEC) is a food-borne pathogen in humans, with cattle being the main reservoir. The objective of this study was to determine the carrying of STEC in Paraguayan bovines and to analyze the virulence profile and serotypes of these isolates. A total of 197 samples of bovine fecal samples and an average of 5 to 50 colonies from stx1/stx2 positive samples were studied. The stx1, stx2, saa, ehxA and eae genes were amplified by PCR. 84.8% of the cattle were carriers of STEC. The predominant virulence profiles were stx2 and stx2/saa/ehxA. The serotyping was performed by agglutination reactions for 60 selected isolates, resulting in isolation of serogroup O103, which could produce infections in humans. This work shows the first data of STEC carriers in Paraguayan cattle, and indicates the need for other studies with greater territorial coverage for a complete vision of this phenomenon.


Assuntos
Escherichia coli Shiga Toxigênica/isolamento & purificação , Criação de Animais Domésticos , Animais , Bovinos , Fezes/microbiologia , Paraguai , Sorotipagem , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/patogenicidade
4.
Rev Argent Microbiol ; 51(3): 208-213, 2019.
Artigo em Espanhol | MEDLINE | ID: mdl-30665746

RESUMO

The ability to form biofilms of pathogenic microorganisms in a wide variety of environments, surfaces and conditions constitute an important risk, both for the food industry and for public health. The aim of this work was to evaluate and to compare the effects of the methodology applied and the culture medium used on the ability of a non-O157 verotoxigenic Escherichia coli strain and an enteropathogenic strain to form biofilm on polystyrene surface. Two methodological variants were tested in static culture and culture mediums with different composition were used. The results showed that both strains were able to form a greater biofilm under culture in LB supplemented with glucose, with medium replacement at 24h and the quantification of the biofilm carried out at 48h of incubation. These conditions could be used in future studies on biofilm formation.


Assuntos
Biofilmes/efeitos dos fármacos , Meios de Cultura/farmacologia , Escherichia coli Enteropatogênica/efeitos dos fármacos , Escherichia coli Shiga Toxigênica/efeitos dos fármacos , Técnicas Bacteriológicas , Biofilmes/crescimento & desenvolvimento , Escherichia coli Enteropatogênica/patogenicidade , Escherichia coli Enteropatogênica/fisiologia , Glucose/farmacologia , Poliestirenos , Escherichia coli Shiga Toxigênica/patogenicidade , Escherichia coli Shiga Toxigênica/fisiologia , Especificidade da Espécie
5.
Rev Argent Microbiol ; 49(2): 174-177, 2017.
Artigo em Espanhol | MEDLINE | ID: mdl-28343858

RESUMO

The genus Lactobacillus daily generates a growing interest among microbiologists and technologists, who try to discover new biotechnological applications and probiotic properties. The main goal of this study was to evaluate the inhibitory capacity of Lactobacillus spp. against pathogens (Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus) involved in foodborne diseases. For this purpose, samples were collected at different stages of the pork production chain. Seventy eight bacterial strains were isolated. Twenty seven (27) of these strains (37.18%) had genotypic and phenotypic characteristics corresponding to Lactobacillus spp. whereas 85.18% of them showed inhibitory capacity. These data showed that the studied strains represent a potential alternative to inactivate foodborne pathogens and thus provide safe food to consumers.


Assuntos
Microbiologia de Alimentos , Doenças Transmitidas por Alimentos , Lactobacillus , Escherichia coli O157 , Doenças Transmitidas por Alimentos/prevenção & controle , Humanos , Probióticos , Salmonella
6.
Foodborne Pathog Dis ; 13(3): 163-70, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26836701

RESUMO

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens, and beef cattle are recognized as the principal reservoir. The aims of this study were (1) to identify the most sensitive combination of selective enrichment broths and agars for STEC isolation in artificially inoculated ground beef samples, and (2) to evaluate the most efficient combination(s) of methods for naturally contaminated ground beef samples. A total of 192 ground beef samples were artificially inoculated with STEC and non-stx bacterial strains. A combination of four enrichment broths and three agars were evaluated for sensitivity, specificity, and positive predictive value for STEC isolation from experimentally inoculated samples. Enrichments with either modified tryptic soy broth (mTSB) containing 8 mg/L novobiocin (mTSB-8) or modified Escherichia coli (mEC) broth followed by isolation in MacConkey agar were the most sensitive combinations for STEC isolation of artificially inoculated samples. Independently, both enrichments media followed by isolation in MacConkey were used to evaluate ground beef samples from 43 retail stores, yielding 65.1% and 58.1% stx-positive samples by RT-PCR, respectively. No difference was observed in the isolate proportions between these two methods (8/25 [32%] and 8/28 [28.6%]). Identical serotypes and stx genotypes were observed in STEC strains isolated from the same samples by either method. In this study, no single enrichment protocol was sufficient to detect all STEC in artificially inoculated samples and had considerable variation in detection ability with naturally contaminated samples. Moreover, none of the single or combinations of multiple isolation agars used were capable of identifying all STEC serogroups in either artificially inoculated or naturally occurring STEC-contaminated ground beef. Therefore, it may be prudent to conclude that there is no single method or combination of isolation methods capable of identifying all STEC serogroups.


Assuntos
Infecções por Escherichia coli/microbiologia , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Carne Vermelha/microbiologia , Toxina Shiga/genética , Escherichia coli Shiga Toxigênica/isolamento & purificação , Ágar , Animais , Bovinos , Meios de Cultura
7.
Rev Argent Microbiol ; 48(4): 325-328, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27667362

RESUMO

Shigatoxigenic Escherichia coli (STEC) is a foodborne pathogen that causes hemolytic uremic syndrome (HUS) and the consumption of chicken products has been related to some HUS cases. We performed a non-selective isolation and characterization of STEC strains from retail chicken products. STEC isolates were characterized according to the presence of stx1, stx2, eae, saa and ehxA; stx subtypes and serotypes. Most of them carried stx2, showing subtypes associated with severe human disease. Although reported in other avian species, the stx2f subtype was not detected. The isolates corresponded to different serotypes and some of them, such as O22:H8, O113:H21, O130:H11, O171:H2 and O178:H19, have also been identified among STEC isolated from patients suffering from diarrhea, hemorrhagic colitis, HUS, as well as from cattle. Considering the virulence profiles and serotypes identified, our results indicate that raw chicken products, especially hamburgers sold at butcheries, can be vehicles for high-risk STEC strains.


Assuntos
Microbiologia de Alimentos , Genes Bacterianos , Produtos Avícolas/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Argentina , Técnicas de Tipagem Bacteriana , Galinhas/microbiologia , Proteínas de Escherichia coli/genética , Sorotipagem , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/patogenicidade , Virulência
8.
Foodborne Pathog Dis ; 12(8): 704-11, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26217917

RESUMO

Enterotoxigenic Escherichia coli (ETEC) is the major pathogen responsible for neonatal diarrhea, postweaning diarrhea, and edema disease in pigs. Although it can be harmless, ETEC is also present in the intestines of other animal species and humans, causing occasional diarrhea outbreaks. The evaluation of this pathogen's presence in food sources is becoming an increasingly important issue in human health. In order to determine the prevalence of ETEC in nondiarrheic pigs, 990 animals from 11 pig farms were sampled. Using end-time polymerase chain reaction (PCR), eltA, estI genes, or both, were detected in 150 (15.2%) animals. From the positive samples, 40 (26.6%) ETEC strains were isolated, showing 19 antibiotic-resistance patterns; 52.5% of these strains had multiple antibiotic resistances, and 17.5% carried the intI2 gene. The most prevalent genotypes were rfb(O157)/estII/aidA (32.5%) and estI/estII (25.0%). The estII gene was identified most frequently (97.5%), followed by estI (37.5%), astA (20.0%), and eltA (12.5%). The genes coding the fimbriae F5, F6, and F18 were detected in three single isolates. The aidA gene was detected in 20 ETEC strains associated with the estII gene. Among the isolated ETEC strains, stx(2e)/estI, stx(2e)/estI/estII, and stx(2e)/estI/estII/intI2 genotypes were identified. The ETEC belonged to 12 different serogroups; 37.5% of them belonged to serotype O157:H19. Isolates were grouped by enterobacterial repetitive intergenic consensus-PCR into 5 clusters with 100.0% similarity. In this study, we demonstrated that numerous ETEC genotypes cohabit and circulate in swine populations without clinical manifestation of neonatal diarrhea, postweaning diarrhea, or edema disease in different production stages. The information generated is important not only for diagnostic and epidemiological purposes, but also for understanding the dynamics and ecology of ETEC in pigs in different production stages that can be potentially transmitted to humans from food animals.


Assuntos
Anti-Infecciosos/análise , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli Enterotoxigênica/isolamento & purificação , Genes Bacterianos , Carne Vermelha/microbiologia , Animais , Anti-Infecciosos/farmacologia , DNA Bacteriano/genética , Diarreia/microbiologia , Diarreia/veterinária , Edematose Suína/microbiologia , Escherichia coli Enterotoxigênica/efeitos dos fármacos , Escherichia coli Enterotoxigênica/genética , Proteínas de Escherichia coli/genética , Contaminação de Alimentos , Microbiologia de Alimentos , Técnicas de Genotipagem , Carne Vermelha/análise , Suínos , Doenças dos Suínos/microbiologia
9.
Rev Argent Microbiol ; 46(2): 126-32, 2014.
Artigo em Espanhol | MEDLINE | ID: mdl-25011597

RESUMO

Verotoxin-producing Escherichia coli (VTEC) is the etiologic agent of hemolytic-uremic syndrome (HUS), which typically affects children ranging in age from six months to five years old. Transmission is produced by consumption of contaminated food, by direct contact with animals or the environment and from person to person. In previous studies we determined that the environment of a dairy farm is a non-animal reservoir; thus, we proposed to study the survival of 4 VTEC isolates (O20:H19; O91:H21; O157:H7 and O178:H19) in sterile water troughs and bovine feces by viable bacteria count and detection of virulence genes by PCR. It was demonstrated that the survival of different VTEC isolates (O157 and non-O157) varied in terms of their own characteristics as well as of the environmental conditions where they were found. The main differences between isolates were their survival time and the maximal counts reached. The competitive and adaptive characteristics of some isolates increase the infection risk for people that are visiting or working on a farm, as well as the risk for reinfection of the animals and food contamination.


Assuntos
Fezes/microbiologia , Escherichia coli Shiga Toxigênica/fisiologia , Microbiologia da Água , Animais , Bovinos
10.
Rev Argent Microbiol ; 46(2): 122-5, 2014.
Artigo em Espanhol | MEDLINE | ID: mdl-25011596

RESUMO

In Argentina, EPEC is one of the most prevalent agents isolated from children with diarrhea. Because contamination with this pathotype could occur during slaughter, the aim of this study was to isolate and characterize EPEC strains obtained from live animals (cloacae), eviscerated carcasses, washed carcasses and water from chillers. Twenty nine isolates of atypical EPEC were characterized. These isolates presented a wide variety of serotypes, some of which (O2:H40, O8:H19 and O108:H9) had been reported in other animal species. Serotype O45:H8, previously isolated from children with diarrhea was also found. Isolates of serotypes O2:H40, O108:H9 and O123:H32 were detected at different stages of the slaughtering process, suggesting that the process is not adequately performed. This latter fact highlights the importance of reinforcing control and hygienic measures at different stages of the chicken slaughtering process in order to reduce microbial contamination.


Assuntos
Galinhas/microbiologia , Escherichia coli Enteropatogênica/isolamento & purificação , Matadouros , Animais
11.
Foods ; 12(14)2023 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-37509768

RESUMO

The objective of this work was to molecularly and genotypically characterize and test the inhibitory activity of six colicinogenic Escherichia coli strains (ColEc) and their partially purified colicins against STEC O157:H7 isolated from clinical human cases. Inhibition tests demonstrated the activity of these strains and their colicins against STEC O157:H7. By PCR it was possible to detect colicins Ia, E7, and B and microcins M, H47, C7, and J25. By genome sequencing of two selected ColEc strains, it was possible to identify additional colicins such as E1 and Ib. No genes coding for stx1 and stx2 were detected after analyzing the genome sequence. The inhibitory activity of ColEc against STEC O157:H7 used as an indicator showed that colicins are potent growth inhibitors of E. coli O157:H7, being a potential alternative to reduce the presence of pathogens of public health relevance.

12.
Curr Microbiol ; 65(3): 337-43, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22706777

RESUMO

Environmental samples were taken from ground, cattle water troughs, and feeders from a dairy farm with different STEC prevalence between animal categories (weaning calves, rearing calves, and dairy cows). Overall, 23 % of samples were positive for stx genes, stx(2) being the most prevalent type. Isolates were analyzed by PCR monoplex to confirm generic E. coli and by two multiplex PCR to investigate the presence of stx(1), stx(2), eae, saa, ehxA, and other putative virulence genes encoded in STEC plasmids: katP, espP, subA, and stcE. The toxin genes were subtyped and the strains were serotyped. The ground and the environment of the rearing calves were the sites with the highest number of STEC-positive samples; however, cattle water troughs and the environment of cows were the places with the greater chance of finding stx(2EDL933) which is a subtype associated with serious disease in humans. Several non-O157 STEC serotypes were detected. The serotypes O8:H19; O26:H11; O26:H-; O118:H2; O141:H-; and O145:H- have been asociated with human illness. Furthermore, the emergent pathogen STEC O157:H- (stx(1)-ehxA-eae) was detected in the environment of the weaning calves. These results emphasize the risk that represents the environment as source of STEC, a potential pathogen for human and suggest the importance of developing control methods designed to prevent contaminations of food products and transmission from animal to person.


Assuntos
Indústria de Laticínios , Microbiologia Ambiental , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Bovinos , Distribuição de Qui-Quadrado , Água Potável/virologia , Feminino , Esterco/virologia , Plasmídeos/genética , Toxinas Shiga/genética , Microbiologia do Solo
13.
Rev Argent Microbiol ; 44(4): 312-23, 2012.
Artigo em Espanhol | MEDLINE | ID: mdl-23267631

RESUMO

Verocytotoxigenic Escherichia coli (VTEC) is associated with outbreaks and sporadic cases of hemorrhagic colitis (HC) and hemolytic-uremic syndrome (HUS), the most severe form of these human diseases. In Argentina HUS is endemic, with 500 new cases per year and an incidence of 17/100,000 in children under 5 years of age. VTEC O157:H7 is the most frequently isolated serotype, although there are non-O157 serotypes that have been associated with human disease. VTEC produces verocytotoxins and accessory virulence factors such as intimin, an enterohemolysin and an autoagglutinating protein called Saa. Cattle are VTEC carriers and several studies in Argentina have confirmed that bovines are the main reservoir of serotypes O157:H7 and non-O157, many of them involved in HUS and HC worldwide. Transmission of VTEC to humans occurs through consumption of undercooked meat, vegetables and water contaminated by feces of carriers, person-to-person and contaminated environment contact. Dairy farms can contribute to the risk of VTEC infection in humans through the consumption of raw milk, dairy products, and contaminated meat from dairy cattle and through contamination of the dairy environment. There is wide distribution and high prevalence of VTEC serotypes in dairy cattle in Argentina; therefore, it is important to improve the measures of control and management and to prevent the transmission of VTEC strains among animals, environment and humans.


Assuntos
Indústria de Laticínios , Escherichia coli Enterotoxigênica/isolamento & purificação , Escherichia coli Enterotoxigênica/patogenicidade , Leite/microbiologia , Toxinas Shiga , Animais , Bovinos , Reservatórios de Doenças , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/transmissão , Humanos
14.
PLoS One ; 15(11): e0242317, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33186398

RESUMO

We developed a quantitative microbiological risk assessment (QMRA) of haemolytic uremic syndrome (HUS) associated with Shiga toxin-producing Escherichia coli (STEC)-contaminated beef (intact beef cuts, ground beef and commercial hamburgers) in children under 15 years of age from Argentina. The QMRA was used to characterize STEC prevalence and concentration levels in each product through the Argentinean beef supply chain, including cattle primary production, cattle transport, processing and storage in the abattoir, retail and home preparation, and consumption. Median HUS probability from beef cut, ground beef and commercial hamburger consumption was <10-15, 5.4x10-8 and 3.5x10-8, respectively. The expected average annual number of HUS cases was 0, 28 and 4, respectively. Risk of infection and HUS probability were sensitive to the type of abattoir, the application or not of Hazard Analysis and Critical Control Points (HACCP) for STEC (HACCP-STEC), stx prevalence in carcasses and trimmings, storage conditions from the abattoir to retailers and home, the joint consumption of salads and beef products, and cooking preference. The QMRA results showed that the probability of HUS was higher if beef cuts (1.7x) and ground beef (1.2x) were from carcasses provided by abattoirs not applying HACCP-STEC. Thus, the use of a single sanitary standard that included the application of HACCP-STEC in all Argentinean abattoirs would greatly reduce HUS incidence. The average number of annual HUS cases estimated by the QMRA (n = 32) would explain about 10.0% of cases in children under 15 years per year in Argentina. Since other routes of contamination can be involved, including those not related to food, further research on the beef production chain, other food chains, person-to-person transmission and outbreak studies should be conducted to reduce the impact of HUS on the child population of Argentina.


Assuntos
Síndrome Hemolítico-Urêmica/microbiologia , Carne Vermelha/microbiologia , Adulto , Animais , Argentina , Bovinos , Criança , Dieta , Escherichia coli/fisiologia , Feminino , Contaminação de Alimentos , Armazenamento de Alimentos , Humanos , Masculino , Medição de Risco
15.
Open Vet J ; 9(1): 65-73, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31086769

RESUMO

Enterotoxigenic Escherichia coli (ETEC) is the most common and global cause of neonatal calf diarrhea, but there is a little information regarding calf ETEC strains in Argentina. In this study, five ETEC isolates from diarrheic dairy calves (2-10 d old) from Buenos Aires and Cordoba, Argentina were characterized on the basis of virulence gene (VG) pattern, O:H serotyping, hemolytic phenotype, phylogenetic group affiliation, antimicrobial (AM) resistance profile, and presence of integron class 1 and 2. The five isolates were examined by polymerase chain reaction (PCR) for the presence of 18 bovine VGs and showed the following genotypes: F5+/F41+/sta+ (D242), F5+/sta+ (D158), F5+/sta+ (D157), F5+ (D151-9), and F5+/iucD+ (D151-5). These VGs confer pathogenic potential and most of them are associated with the ETEC pathotype. The five isolates showed a non-hemolytic phenotype, belonged to five different serotypes: O101:H-, O141:H-, O60:H-, ONT:H10, and ONT:H-, and were assigned to the phylogenetic group A by the quadruplex Clermont PCR method. The AM resistance of the three isolates D242, D157, and D151-5 was determined by agar disk diffusion method for 24 AMs and they exhibited a multi-resistance phenotype (resistance to four different AM classes: Cephalosporins, Penicillins, Macrolides, and Ansamycins). In addition, class 1 integrons were found in the isolate D151-5 containing the dfrA17-aadA5 gene cassette and in the bovine ETEC reference strain FV10191 containing the dfrA1-aadA1 gene cassette. The present study revealed for the first time the occurrence of multi-resistant ETEC associated with neonatal diarrhea in dairy calves in Argentina. This finding may be used for diagnostic and therapeutic purposes.


Assuntos
Doenças dos Bovinos/microbiologia , Farmacorresistência Bacteriana/genética , Escherichia coli Enterotoxigênica/fisiologia , Escherichia coli Enterotoxigênica/patogenicidade , Infecções por Escherichia coli/veterinária , Animais , Animais Recém-Nascidos , Antibacterianos/farmacologia , Argentina , Bovinos , Escherichia coli Enterotoxigênica/efeitos dos fármacos , Escherichia coli Enterotoxigênica/genética , Infecções por Escherichia coli/microbiologia , Integrons/efeitos dos fármacos , Integrons/genética , Fenótipo , Sorotipagem/veterinária , Virulência
16.
Microorganisms ; 6(4)2018 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-30274159

RESUMO

The aim of this study was to investigate the presence of class 1 integrons in a collection of Shiga toxin-producing Escherichia coli (STEC) from different origins and to characterize pheno- and genotypically the antimicrobial resistance associated to them. A collection of 649 isolates were screened for the class 1 integrase gene (intI1) by Polymerase chain reaction The variable region of class 1 integrons was amplified and sequenced. Positive strains were evaluated for the presence of antimicrobial resistance genes with microarray and for antimicrobial susceptibility by the disk diffusion method. Seven out of 649 STEC strains some to serogroups, O26, O103 and O130 isolated from cattle, chicken burger, farm environment and pigs were identified as positive for intl1. Different arrangements of gene cassettes were detected in the variable region of class 1 integron: dfrA16, aadA23 and dfrA1-aadA1. In almost all strains, phenotypic resistance to streptomycin, tetracycline, trimethoprim/sulfamethoxazole, and sulfisoxazole was observed. Microarray analyses showed that most of the isolates carried four or more antimicrobial resistance markers and STEC strains were categorized as Multridrug-resistant. Although antimicrobials are not usually used in the treatment of STEC infections, the presence of Multridrug-resistant in isolates collected from farm and food represents a risk for animal and human health.

17.
Artigo em Inglês | MEDLINE | ID: mdl-28503491

RESUMO

Shiga toxin-producing Escherichia coli (STEC) are pathogens of significant public health concern. Several studies have confirmed that cattle are the main reservoir of STEC in Argentina and other countries. Although Shiga toxins represent the primary virulence factors of STEC, the adherence and colonization of the gut are also important in the pathogenesis of the bacteria. The aim of this study was to analyze and to compare the presence of putative virulence factors codified in plasmid -katP, espP, subA, stcE- and adhesins involved in colonization of cattle -efa1, iha- in 255 native STEC strains isolated from different categories of cattle from different production systems. The most prevalent gene in all strains was espP, and the less prevalent was stcE. katP was highly detected in strains isolated from young and rearing calves (33.3%), while subA was predominant in those isolated from adults (71.21%). Strains from young calves showed the highest percentage of efa1 (72.46%), while iha showed a high distribution in strains from rearing calves and adults (87.04 and 98.48% respectively). It was observed that espP and iha were widely distributed throughout all strains, whereas katP, stcE, and efa1 were more associated with the presence of eae and subA with the eae-negative strains. A great proportion of eae-negative strains were isolated from adults -dairy and grazing farms- and from rearing calves -dairy and feedlot-, while mostly of the eae-positive strains were isolated from dairy young calves. Data exposed indicate a correlation between the category of the animal and the production systems with the presence or absence of several genes implicated in adherence and virulence of STEC.


Assuntos
Bovinos/microbiologia , Infecções por Escherichia coli/veterinária , Toxinas Shiga/biossíntese , Escherichia coli Shiga Toxigênica/genética , Fatores de Virulência/genética , Virulência/genética , Adesinas Bacterianas/genética , Animais , Argentina , Toxinas Bacterianas/genética , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Proteína Reguladora de Resposta a Leucina/genética , Metaloendopeptidases/genética , Plasmídeos/genética , Serina Endopeptidases/genética , Escherichia coli Shiga Toxigênica/crescimento & desenvolvimento , Escherichia coli Shiga Toxigênica/isolamento & purificação , Escherichia coli Shiga Toxigênica/patogenicidade , Subtilisinas/genética
18.
Sci Rep ; 7(1): 7011, 2017 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-28765569

RESUMO

Shiga Toxin-producing Escherichia coli (STEC) are a group of foodborne pathogens associated with diarrhea, dysentery, hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). Shiga toxins are the major virulence factor of these pathogens, however adhesion and colonization to the human intestine is required for STEC pathogenesis. A subset of STEC strains carry the Locus of Enterocyte Effacement (LEE) pathogenicity island (PAI), which encodes genes that mediate the colonization of the human intestine. While LEE-positive STEC strains have traditionally been associated with human disease, the burden of disease caused by STEC strains that lacks LEE (LEE-negative) has increased recently in several countries; however, in the absence of LEE, the molecular pathogenic mechanisms by STEC strains are unknown. Here we report a 86-kb mosaic PAI composed of four modules that encode 80 genes, including novel and known virulence factors associated with adherence and autoaggregation. Therefore, we named this PAI as Locus of Adhesion and Autoaggregation (LAA). Phylogenomic analysis using whole-genome sequences of STEC strains available in the NCBI database indicates that LAA PAI is exclusively present in a subset of emerging LEE-negative STEC strains, including strains isolated from HC and HUS cases. We suggest that the acquisition of this PAI is a recent evolutionary event, which may contribute to the emergence of these STEC.


Assuntos
Adesinas Bacterianas/genética , Infecções por Escherichia coli/microbiologia , Loci Gênicos , Ilhas Genômicas , Escherichia coli Shiga Toxigênica/genética , Fatores de Virulência/genética , Biologia Computacional , Genoma Bacteriano , Humanos , Filogenia , Escherichia coli Shiga Toxigênica/classificação
19.
J Med Microbiol ; 55(Pt 5): 545-549, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16585641

RESUMO

Most cases of diarrhoea-associated haemolytic uraemic syndrome (HUS) are caused by verocytotoxin-producing Escherichia coli (VTEC). Argentina has the highest worldwide incidence of HUS, but with a lower incidence of VTEC O157 : H7 serotype than non-Latin American countries. A large number of VTEC serotypes have been isolated from cattle and cattle-derived food products in Argentina. The aim of this work was to study intraserotype genetic diversity among these VTEC strains by random amplification of polymorphic DNA (RAPD). Strains were selected that belonged to the same serotype, but had been isolated from different sources (cattle and meat). Intraserotype genetic diversity was detected among strains belonging to O20 : H19, O113 : H21, O117 : H7, O157 : H7, O171 : H2 and O174 : H21, but only one RAPD profile corresponded to strains belonging to O91 : H21, although these isolates were from different sources.


Assuntos
Infecções por Escherichia coli/veterinária , Escherichia coli O157/classificação , Escherichia coli/classificação , Polimorfismo Genético , Toxinas Shiga/biossíntese , Animais , Antígenos de Bactérias/análise , Argentina , Bovinos , Doenças dos Bovinos/microbiologia , Impressões Digitais de DNA , DNA Bacteriano/análise , DNA Bacteriano/genética , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Escherichia coli/fisiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/isolamento & purificação , Escherichia coli O157/patogenicidade , Escherichia coli O157/fisiologia , Variação Genética , Carne/microbiologia , Antígenos O/análise , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sorotipagem
20.
Front Microbiol ; 7: 93, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26903972

RESUMO

Pigs are important reservoirs of Shiga toxin-producing Escherichia coli (STEC). The entrance of these strains into the food chain implies a risk to consumers because of the severity of hemolytic uremic syndrome. This study reports the prevalence and characterization of STEC throughout the pork production chain. From 764 samples, 31 (4.05%) were stx positive by PCR screening. At farms, 2.86% of samples were stx positive; at slaughter, 4.08% of carcasses were stx positive and at boning rooms, 6% of samples were stx positive. These percentages decreased in pork meat ready for sale at sales markets (4.59%). From positive samples, 50 isolates could be characterized. At farms 37.5% of the isolates carried stx1/stx2 genes, 37.5% possessed stx2e and 25%, carried only stx2. At slaughter we detected 50% of isolates positive for stx2, 33% for stx2e, and 16% for stx1/stx2. At boning rooms 59% of the isolates carried stx1/stx2, 14% stx2e, and 5% stx1/stx2/stx2e. At retail markets 66% of isolates were positive for stx2, 17% stx2e, and 17% stx1/stx2. For the other virulence factors, ehxA and saa were not detected and eae gene was detected in 12% of the isolates. Concerning putative adhesins, agn43 was detected in 72%, ehaA in 26%, aida in 8%, and iha in 6% of isolates. The strains were typed into 14 E. coli O groups (O1, O2, O8, O15, O20, O35, O69, O78, O91, O121, O138, O142, O157, O180) and 10 H groups (H9, H10, H16, H21, H26, H29, H30, H32, H45, H46). This study reports the prevalence and characterization of STEC strains through the chain pork suggesting the vertical transmission. STEC contamination originates in the farms and is transferred from pigs to carcasses in the slaughter process and increase in meat pork at boning rooms and sales markets. These results highlight the need to implement an integrated STEC control system based on good management practices on the farm and critical control point systems in the food chain.

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