Detection and analysis of Shiga toxin producing and enteropathogenic Escherichia coli in cattle from Tierra del Fuego, Argentina.

Shiga toxin producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) are pathovars that affect mainly infants' health. Cattle are the main reservoir of STEC. Uremic hemolytic syndrome and diarrheas can be found at high rates in Tierra del Fuego (TDF). This study aimed to establish the prevalence of STEC and EPEC in cattle at slaughterhouses in TDF and to analyze the isolated strains. Out of 194 samples from two slaughterhouses, STEC prevalence was 15%, and EPEC prevalence was 5%. Twenty-seven STEC strains and one EPEC were isolated. The most prevalent STEC serotypes were O185:H19 (7), O185:H7 (6), and O178:H19 (5). There were no STEC eae + strains (AE-STEC) or serogroup O157 detected in this study. The prevalent genotype was stx2c (10/27) followed by stx1a/stx2hb (4/27). Fourteen percent of the strains presented at least one stx non-typeable subtype (4/27). Shiga toxin production was detected in 25/27 STEC strains. The prevalent module for the Locus of Adhesion and Autoaggregation (LAA) island was module III (7/27). EPEC strain was categorized as atypical and with the ability to cause A/E lesion. The ehxA gene was present in 16/28 strains, 12 of which were capable of producing hemolysis. No hybrid strains were detected in this work. Antimicrobial susceptibility tests showed that all strains were resistant to ampicillin and 20/28 were resistant to aminoglycosides. No statistical differences could be seen in the detection of STEC or EPEC either by slaughterhouse location or by production system (extensive grass or feedlot). The rate of STEC detection was lower than the one reported for the rest of Argentina. STEC/EPEC relation was 3 to 1. This is the first study on cattle from TDF as reservoir for strains that are potentially pathogenic to humans.

Potential Zoonotic Pathovars of Diarrheagenic Escherichia coli Detected in Lambs for Human Consumption from Tierra del Fuego, Argentina.

Diarrheagenic Escherichia coli (DEC) pathovars impact childhood health. The southern region of Argentina shows the highest incidence of hemolytic uremic syndrome (HUS) in children of the country. The big island of Tierra del Fuego (TDF) in Argentina registered an incidence of five cases/100,000 inhabitants of HUS in 2019. This work aimed to establish the prevalence of STEC, EPEC, and EAEC in lambs slaughtered in abattoirs from TDF as well as to characterize the phenotypes and the genotypes of the isolated pathogens. The prevalence was 26.6% for stx+, 5.7% for eae+, and 0.27% for aagR+/aaiC+. Twelve STEC isolates were obtained and belonged to the following serotypes: O70:HNT, O81:H21, O81:HNT, O102:H6, O128ab:H2, O174:H8, and O174:HNT. Their genotypic profiles were stx1c (2), stx1c/ehxA (3), stx2b/ehxA (1), stx1c/stx2b (2), and stx1c/stx2/ehxA (4). Six EPEC isolates were obtained and corresponded to five serotypes: O2:H40, O32:H8, O56:H6, O108:H21, and O177:H25. All the EPEC isolates were bfpA- and two were ehxA+. By XbaI-PFGE of 17 isolates, two clusters were identified. By antimicrobial susceptibility tests, 8/12 STEC and 5/6 EPEC were resistant to at least one antibiotic. This work provides new data to understand the ecology of DEC in TDF and confirms that ovine are an important carrier of these pathogens in the region.

Caracterización de cepas patógenas extraintestinales de Escherichia coli aisladas de perros y gatos de companía de Buenos Aires, Argentina / Characterization of extraintestinal pathogenic Escherichia coli strains isolated from household dogs and cats in Buenos Aires, Argentina

El pangenoma de Escherichia coli se compone de un core conservado y regiones genómicas variables. El componente genético constante permite determinar la filogenia del microorganismo, mientras que la variabilidad genética ha permitido el surgimiento de cepas patógenas intestinales y extraintestinales. En este estudio caracterizamos genéticamente 85 cepas patógenas extraintestinales aisladas de caninos y felinos. Se utilizó el esquema de Clermont para agruparlas en relación con elfilogrupo de pertenencia (intestinal: Ay B1; extraintestinal: B2 y D) y se investigó en ellas la presencia de varios marcadores de virulencia (pap1-2, pap3-4, sfa, afa, hlyA, aer y cnf), así como de marcadores de patovares híbridos. El 69,4% de los aislamientos pertenecieron al filogrupo A; el 1,2% al B1; el 16,5% al B2 y el 12,9% al D. El gen encontrado con mayor frecuencia fue sfa (21/85), seguido de los genes pap1-2 y cnf (20/85) y pap3-4 (19/85). No se detectaron híbridos. Los aislamientos en animales deben ser estudiados debido al potencial zoonótico del microorganismo.

The pangenome of Escherichia coli is composed of a conserved core and variable genomic regions. The constant genetic component allows to determine the phylogeny of the microorganism, while genetic variability promoted the emergence of intestinal pathogenic strains and extraintestinal strains. In this study we characterized 85 extraintestinal pathogenic isolates genetically isolated from canines and felines. We used the Clermont scheme that includes intestinal (A and B1) and extraintestinal (B2 and D) phylogroups, virulence markers (pap1-2, pap3-4, sfa, afa, hlyA, aer and cnf) and hybrid pathogens. A percentage of 69.4% of the isolates belonged to phylogroupA; 1.2% to phylogroup B1; 16.5% to phylogroup B2 and 12.9% to phylogroup D. The most commonly found gene was sfa (21/85), followed by pap1-2 and cnf (20/85) and pap3-4 (19/85). No hybrids were detected. Animal isolates should be studied due to the zoonotic potential of the microorganism.

[Characterization of extraintestinal pathogenic Escherichia coli strains isolated from household dogs and cats in Buenos Aires, Argentina]. / Caracterización de cepas patógenas extraintestinales de Escherichia coli aisladas de perros y gatos de compañía de Buenos Aires, Argentina.

The pangenome of Escherichia coli is composed of a conserved core and variable genomic regions. The constant genetic component allows to determine the phylogeny of the microorganism, while genetic variability promoted the emergence of intestinal pathogenic strains and extraintestinal strains. In this study we characterized 85 extraintestinal pathogenic isolates genetically isolated from canines and felines. We used the Clermont scheme that includes intestinal (A and B1) and extraintestinal (B2 and D) phylogroups, virulence markers (pap1-2, pap3-4, sfa, afa, hlyA, aer and cnf) and hybrid pathogens. A percentage of 69.4% of the isolates belonged to phylogroup A; 1.2% to phylogroup B1; 16.5% to phylogroup B2 and 12.9% to phylogroup D. The most commonly found gene was sfa (21/85), followed by pap1-2 and cnf (20/85) and pap3-4 (19/85). No hybrids were detected. Animal isolates should be studied due to the zoonotic potential of the microorganism.

Detection and Characterization of Enteropathogenic and Shiga Toxin-Producing Escherichia coli Strains in Rattus spp. from Buenos Aires.

Enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) are pathovars of E. coli that impact human health by causing childhood diseases. In this work, 118 synanthropic rodents of the genus Rattus from Buenos Aires, Argentina were evaluated as EPEC and STEC carriers. Rectal swab samples from captured animals were evaluated by conventional PCR to detect the presence of the eae, stx1, stx2, and rfbO157 genes. Twenty-one isolates were obtained (17 EPEC isolates from seven animals and four STEC isolates from the same animal). All EPEC isolates tested negative for the presence of the bfpA gene. One EPEC isolate carried the iha gene, and five EPEC isolates carried the toxB gene. STEC isolates exhibited two different virulence profiles: stx1a/stx2a/stx2c/stx2d/saa/ehxA/subA (3/4) and stx1a/stx2a/saa/ehxA/subA (1/4). EPEC isolate serotypes included O109:H46 (7), O71:H40 (4), O71:NM (2), O138:H40 (1), O108:H21 (1), O88:H25 (1), and O76:NM (1), and STEC isolates belonged to the O108:H11 (4) serotype. Antimicrobial susceptibility testing was carried out, and resistance to tetracycline was observed in one EPEC strain. Our results demonstrate that Rattus spp. may act as carriers of EPEC and STEC strains and may be involved in the epidemiology of diarrheal disease in infancy.

Putative virulence factors and clonal relationship of O174 Shiga toxin-producing Escherichia coli isolated from human, food and animal sources.

Shiga toxin-producing Escherichia coli (STEC) is th etiological agent of gastrointestinal diseases as haemorrhagic colitis and haemolytic-uraemic syndrome (HUS). Shiga toxin (Stx) is the main virulence factor. There are two types, Stx1 and Stx2, and several subtypes. Other virulence factors are involved in pathogenesis. While O157:H7 is the predominant serotype, at present non-O157 STEC strains are increasingly recognized as foodborne pathogens worldwide. In Argentina, STEC O174 stands out as an emerging pathogen and is one of the four most prevalent non-O157 STEC serogroups. The aim of this study was to characterize 41 O174:[H21, H28] STEC strains isolated from animals, food, and humans. Isolates were characterized by stx genotyping, adhesion factors (afaC, eae, iha, lpfO113, saa, and toxB), additional toxins (cdtV, ehxA, subA) and clonal relationship by pulsed-field gel electrophoresis (PFGE). Among 30 O174:H21 strains, the most prevalent stx subtype was stx2c (56.7%), and among 11 O174:H28 strains, the most prevalent was stx2a (90.9%). Regarding virulence factors, all strains were positive for afaC gene and negative for eae and toxB genes. In O174:H21, the frequency of additional factors was lpfO113 (96.6%), iha (83.3%), ehxA and saa (10%), and subA (3.3%), meanwhile in O174:H28 strains the frequency was iha and subA (100%), lpfO113 (90.9%), ehxA and saa (90.9%), and cdtV (9.09%). By Xbal-PFGE, 29 patterns were established with 64.01% similarity and three clusters were detected. Given the fact that the O174 serogroup is a local emergent, it is important to study the virulence profiles to understand its potential pathogenicity.

Erysipelothrix rhusiopathiae isolated from gull-inflicted wounds in southern right whale calves.

Southern right whales Eubalaena australis from Peninsula Valdés, Argentina, show wounds produced by kelp gulls Larus dominicanus that feed on the whales' dorsal skin and blubber. During the 2013 whale season, several calves were reported showing kelp gull injuries with a swollen area surrounded by rhomboid-shaped raised edges. Samples from 9 calves were taken in order to establish the etiology of these rhomboid-shaped wounds; 2 calves (one living, one dead) showed gull-inflicted injuries with rhomboid-shaped edges. Samples from the dead calf were histologically characterized by the presence of dermal congestion, suppurative dermatitis and panniculitis, necrotizing vasculitis and vascular thrombosis. Erysipelothrix rhusiopathiae was detected by culture and PCR in samples from both calves. In this study we report, for the first time to our knowledge, the isolation of E. rhusiopathiae from wounds produced by gull attacks on southern right whale calves, supplying evidence that these wounds may act as an entry route for pathogens. This work provides new information about the consequences of gull-inflicted injuries for whale health.

Molecular identification of coagulase-negative staphylococci isolated from bovine mastitis and detection of ß-lactam resistance.

INTRODUCTION: Bovine mastitis is a frequent cause of economic loss in dairy herds. Coagulase-negative staphylococci (CoNS) are increasing in importance as causeof bovine intramammary infection (IMI) throughout the world in recent years. CoNShave been isolated from milk samples collected from cows with clinical andsubclinical mastitis in several countries. Identification of mastitis pathogensis important when selection appropriate antimicrobial therapy. METHODOLOGY: A total of 93 strains of Staphylococcusspp isolated from bovine clinical andsubclinical mastitis in Argentina during 2010-2013 were identified by Restriction Fragment Length Polymorphism (RFLP)-PCR using the gap gene. The isolates were tested by PCR for the presence of blaZ gene and mecA gene and were tested by disk diffusion for the susceptibilityto penicillin and cefoxitin. RESULTS: The most common CoNS species was S.chromogenes 46.2% (43/93), followed by S. devriesei 11.8% (11/93) and S. haemolyticus 9.7% (9/93). The blaZ gene was detected in 19 (20.4%) but only 16 (17.2%) isolates were resistant to penicillin; the mecA was detected in6 (6.5%) isolates but only 4 (4.3) were resistant to cefoxitin. The 6 mecA-positive isolates showed oxaxillinMICs ≤ 0.5 µg/ml. DISCUSSION: CoNSare important minor mastitis pathogens and can be the cause of substantial economic losses. Despite the low resistance to PEN in Argentina, the presenceof MR isolates found in this study emphasize the importance of identificationof CoNS when an IMI is present because of the potentially risk of lateraltransfer of resistance genes between staphylococcal species.

Clade 8 and Clade 6 Strains of Escherichia coli O157:H7 from Cattle in Argentina have Hypervirulent-Like Phenotypes.

The hemolytic uremic syndrome (HUS) whose main causative agent is enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a disease that mainly affects children under 5 years of age. Argentina is the country with the highest incidence of HUS in the world. Cattle are a major reservoir and source of infection with E. coli O157:H7. To date, the epidemiological factors that contribute to its prevalence are poorly understood. Single nucleotide polymorphism (SNP) typing has helped to define nine E. coli O157:H7 clades and the clade 8 strains were associated with most of the cases of severe disease. In this study, eight randomly selected isolates of EHEC O157:H7 from cattle in Argentina were studied as well as two human isolates. Four of them were classified as clade 8 through the screening for 23 SNPs; the two human isolates grouped in this clade as well, while two strains were closely related to strains representing clade 6. To assess the pathogenicity of these strains, we assayed correlates of virulence. Shiga toxin production was determined by an ELISA kit. Four strains were high producers and one of these strains that belonged to a novel genotype showed high verocytotoxic activity in cultured cells. Also, these clade 8 and 6 strains showed high RBC lysis and adherence to epithelial cells. One of the clade 6 strains showed stronger inhibition of normal water absorption than E. coli O157:H7 EDL933 in human colonic explants. In addition, two of the strains showing high levels of Stx2 production and RBC lysis activity were associated with lethality and uremia in a mouse model. Consequently, circulation of such strains in cattle may partially contribute to the high incidence of HUS in Argentina.

Emerging types of Shiga toxin-producing E. coli (STEC) O178 present in cattle, deer, and humans from Argentina and Germany.

More than 400 serotypes of Shiga toxin-producing Escherichia coli (STEC) have been implicated in outbreaks and sporadic human diseases. In recent years STEC strains belonging to serogroup O178 have been commonly isolated from cattle and food of bovine origin in South America and Europe. In order to explore the significance of these STEC strains as potential human pathogens, 74 German and Argentinean E. coli O178 strains from animals, food and humans were characterized phenotypically and investigated for their serotypes, stx-genotypes and 43 virulence-associated markers by a real-time PCR-microarray. The majority (n = 66) of the O178 strains belonged to serotype O178:H19. The remaining strains divided into O178:H7 (n = 6), O178:H10 (n = 1), and O178:H16 (n = 1). STEC O178:H19 strains were mainly isolated from cattle and food of bovine origin, but one strain was from a patient with hemolytic uremic syndrome (HUS). Genotyping of the STEC O178:H19 strains by pulsed-field gel electrophoresis revealed two major clusters of genetically highly related strains which differ in their stx-genotypes and non-Stx putative virulence traits, including adhesins, toxins, and serine-proteases. Cluster A-strains including the HUS-strain (n = 35) carried genes associated with severe disease in humans (stx2a, stx2d, ehxA, saa, subAB1, lpfAO113 , terE combined with stx1a, espP, iha). Cluster B-strains (n = 26) showed a limited repertoire of virulence genes (stx2c, pagC, lpfAO113 , espP, iha). Among O178:H7 strains isolated from deer meat and patients with uncomplicated disease a new STEC variant was detected that is associated with the genotype stx1c/stx2b/ehxA/subAB2/espI/[terE]/espP/iha. None of the STEC O178 strains was positive for locus of enterocyte effacement (LEE)- and nle-genes. Results indicate that STEC O178:H19 strains belong to the growing group of LEE-negative STEC that should be considered with respect to their potential to cause diseases in humans.

Characterization of Shiga toxin-producing Escherichia coli isolated from ground beef collected in different socioeconomic strata markets in Buenos Aires, Argentina.

Consumption of raw/undercooked ground beef is the most common route of transmission of Shiga toxin-producing E. coli (STEC). The aim of the study was to determine the STEC contamination level of the ground beef samples collected in 36 markets of different socioeconomic strata in Buenos Aires, Argentina, and the characterization of the isolated strains. Ninety-one out of 252 (36.1%) samples were stx+. Fifty-seven STEC strains were recovered. Eleven STEC strains belonged to O157 serogroup, and 46 to non-O157 serogroups. Virulence markers of the 57 STEC were stx1, 5.3% (3/57); stx2, 86.0% (49/57); stx1/stx2, 8.8% (5/57); ehxA, 61.4% (35/57); eae, 26.3% (15/57); saa, 24.6% (14/57). Shiga toxin subtypes were stx2, 31.5% (17/54); stx2c-vhb, 24.1% (13/54); stx2c-vha, 20.4% (11/54); stx2/stx2c-vha, 14.8% (8/54); stx2/stx2c-vhb, 5.6% (3/54); stx2c-vha/vhb, 3.7% (2/54). Serotypes O178:H19 and O157:H7 were prevalent. Contamination rate of STEC in all strata was high, and the highest O157 contamination was observed at low strata at several sampling rounds. Persistence of STEC was not detected. Sixteen strains (28.1%) were resistant to ampicillin, streptomycin, amikacin, or tetracycline. The STEC contamination level of ground beef could vary according to the sociocultural characteristics of the population.

The icaA gene in staphylococci from bovine mastitis.

INTRODUCTION: Staphylococcus aureus and coagulase-negative staphylococci (CNS) are frequently isolated from cows with mastitis. A main virulence factor of CNS is the ability to adhere and form biofilms. The intercellular gene cluster adhesion (ica) operon is one factor involved in biofilm production although ica-independent factors are also involved. Previous reports based on the results of S. epidermidis and S. aureus suggested that ica is highly conserved between species, but this detection decreases in other CNS biofilm producers. In this study we evaluated the presence of the icaA gene in strains of Staphylococcus spp. isolated from the milk of bovines with mastitis. METHODOLOGY: Thirty-seven staphylococci strains were evaluated by detecting the icaA gene. A new set of PCR primers was designed by consensus region of eight staphylococci from GenBank. Species characterization was performed using the Kloos and Schleifer scheme. RESULTS: We identified the presence of the gene in S. aureus (n:4), S. chromogenes (n:4), and S. sciuri strains (n:2). We also, identified the presence of the gene in S. xylosus (n:5) for the first time. The icaA gene was not detected in S. capitis (n:1), S. epidermidis (n:2), S. hominis (n:2), S. saccharolyticus (n:1), S. simulans (n:4) and S. saprophyticus (n:3). The icaA gene was detected in 40.54% (15/37) of the CNS evaluated. CONCLUSIONS: Our results confirm the presence of the ica operon in various species of CNS pointing to polysaccharide intercellular adhesin (PIA) as the most important component for the formation of biofilms.

Synanthropic rodents as possible reservoirs of shigatoxigenic Escherichia coli strains.

Shigatoxigenic Escherichia coli (STEC) strains are worldwide zoonotic pathogen responsible for different cases of human disease including hemolytic uremic syndrome (HUS). Transmission of STEC to humans occurs through the consumption of food and water contaminated by faeces of carriers and by person-to-person contact. The objective of this study was two-fold: (1) to investigate whether synanthropic rodents are possible reservoirs of STEC in the urban area and (2) whether a particular genus out of synanthropic rodent is the principal carrier of STEC. One hundred and forty-five rodents were captured in Buenos Aires City. Screening for stx1/stx2 and rfbO157 was done by PCR from the confluence zone. STEC isolates were further characterized with biochemical tests by standard methods. Additional virulence factors (eae, ehxA, and saa) were also determined by PCR. Forty-one of the rodents were necropsied and sample of kidney and small and large intestine were taken for histopathological diagnosis. The samples sections were stained with hematoxylin-eosin, and observed by light microscopy to evaluate the systemic involvement of these species in natural infections. STEC was isolated from seven out of 27 suspect animals at screening. The following genotypes were found in the STEC strains: stx1/stx2/ehxA (1), stx2 (4), stx2/ehxA (1), stx2/ehxA/eae (1). Neither gross nor microscopic lesions compatible with those produced by Shiga toxin were observed in the studied organs of necropsied rodents. The bivariate analysis including the 145 rodent's data showed that the isolation of STEC is associated positively to Rattus genus. This synanthropic species may play a role in the transmissibility of the agent thus being a risk to the susceptible population. Their control should be included specifically in actions to dismiss the contamination of food and water by STEC in the urban area, as additional strategies for epidemiological control.

Clonal relations of atypical enteropathogenic Escherichia coli O157:H16 strains isolated from various sources from several countries.

Atypical enteropathogenic Escherichia coli (aEPEC) is comprised of a large heterogeneous group of strains and serotypes that carry the intimin gene (eae) but no other EPEC virulence factors. In a previous study, we examined a few aEPEC strains of O157:H16 serotype from the U.S. and France and found these to be nearly homologous, and speculated that the same strain had been disseminated or perhaps they are part of a large clonal group that exists worldwide. To test that hypothesis, we examined additional 45 strains isolated from various sources from 4 other countries and determined that although there are a few eae-negative O157:H16 strains, most are aEPEC that carried eae and specifically, the ε-eae allele. Analysis by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing showed that as a whole, O157:H16 strains are phylogenetically diverse and have different sequence types and PFGE profiles. But the aEPEC strains within the O157:H16 serotype, regardless of the eae allele carried, are a highly conserved and homologous group of sequence type (ST)-171 strains that shared similar PFGE profiles. These aEPEC strains of O157:H16 serotype are not closely related to any of the major EPEC and enterohemorrhagic E. coli clonal lineages and appear to be part of a large clonal group that are prevalent worldwide.

Escherichia coli shigatoxigénica en animales relacionados con casos de diarreas sanguinolentas o síndrome urémico hemolítico y prevalencia en roedores de la Ciudad de Buenos Aires / Shiga toxin-producing escherichia coli in animals related to cases of bloody diarrhea

Escherichia coli shigatoxigénica (STEC) es un patógeno endémico en Argentina, responsable de diarrea aguda sanguinolenta (DAS) y/o síndrome urémico hemolítico (SUH). La correlación entre SUH y alimentos contaminados ha sido documentada, aunque no siempre se estableció la fuente de infección. La ruta de contagio persona-persona es relevante. Dados los registros previos de prevalencia de STEC en animales de compañía y los hábitos de convivencia humano-animal en centros urbanos, es necesario evaluar la ruta mascota-persona. A su vez, los roedores podrían tener un papel epidemiológico en la endemia. OBJETIVO: Estudiar posibles reservorios animales relacionados con casos de SUH/DAS en la Ciudad Autónoma de Buenos Aires y estimar la prevalencia de STEC en roedores. MÉ-TODOS: Se intervino en 28 casos de SUH y 49 de DAS. Se realizó rastrillaje de cepas STEC por PCR a partir de hisopados rectales de los animales vinculados a cada caso. La prevalencia en roedores se estimó por PCR de sus hisopados rectales. RESULTADOS: Se aislaron cepas STEC en 1/10 caninos y 1/3 felinos con vivientes con casos de SUH, y 1/9 felinos contacto con casos de DAS. Rattus rattus fue hospedero de cepas STEC en 33% de los animales capturados en focos de SUH. En roedores, la prevalencia fue de 3,1%. CONCLUSIONES: Las cepas STEC circulan en los animales que conviven o tienen al menos un hábitat compartido con la población en riesgo, quienes podrían participar en la transmisión del agente. Es necesario reevaluar las intervenciones sanitarias en focos y en programas de control de SUH/DAS

Shigatoxigenic Escherichia coli(STEC) is an endemic pathogen in Argentina, which causes bloody diarrhea (BD) and/or hemolytic uremic syndrome (HUS).The co-relation between HUS and contaminated food has been documented, although the source of infection was not always established. Person-to-person route of infection is relevant. Taking into account previous STEC prevalence data in companion animals and the habits of human-animal coexistence in urban centers, it is necessary to evaluate pet-to-person transmission. On the other hand, rodents may also play an epidemiologic role in the endemic transmission. OBJECTIVE: To study potential animal reservoirs related to HUS and BD cases in the City of Buenos Aires and to estimate the prevalence of STEC in rodents. METHODS: An intervention was conducted in 28 cases of HUS and 49 of BD. Screening for STEC was performed by PCR from rectal swabs of linked animals to each case. The prevalence in rodents was estimated by PCR from rectal swabs. RESULTS: STEC strains were isolated in 1/10 dogs and 1/3 cats cohabiting with HUS cases, and in 1/9 cats in contact with DAS cases. Rattus rattus was host of STEC strains in 33% of the animals captured in HUS areas. In rodents, the prevalence was 3.1%. CONCLUSIONS: STEC strains circulate in animals that live withor share at least the same habitat with the population at risk, and could participate in the transmission of the agent. It is necessary to re-evaluate health interventions both in outbreaks and in control programs of HUS/BD

Risk factors for Shiga toxin-producing Escherichia coli infections in preadolescent schoolchildren in Buenos Aires, Argentina.

INTRODUCTION: Shiga toxin-producing Escherichia coli (STEC) infections are the leading cause of hemolytic uremic syndrome (HUS). STEC is the most common cause of acute kidney disease, responsible for 20% of renal transplants in Argentina. METHODOLOGY: In 2007, an epidemiological survey was conducted among 883 students from the fifth and sixth years of elementary education in the public schools of San Martin City, Buenos Aires, Argentina. Degree of exposure to the known risk factors previously detected in the region as primary causes of STEC infections was evaluated. Risk factors assessed included consumption of hamburgers, poor personal hygiene, and exposure to various types of drinking and recreational water. The study was designed to evaluate exposure to risk factors for STEC infections among different socioeconomic groups. RESULTS: Ninety-five percent of children surveyed reported consumption of hamburgers. Most of these hamburgers were precooked. Children of high and medium strata attended private swimming-pools, while children from the low stratum attended public pools. Only 30.2% of students washed their hands after going to the toilet and only 43.5% reported hand-washing before eating. CONCLUSIONS: Students demonstrated high levels of exposure to identified risk factors for STEC infections. Reduction of these risks will require cultural changes aimed at decreasing morbidity caused by food-borne infections. Institutional framework must provide the necessary resources to implement these changes and emphasize the importance of good personal hygiene. Health education must be implemented to increase food safety awareness of the consumers.

First isolation in Argentina of a highly virulent Shiga toxin-producing Escherichia coli O145:NM from a domestic cat.

INTRODUCTION: Hemolytic uremic syndrome (HUS) is distributed worldwide. In Argentina, more than 450 cases of HUS, mostly sporadic, are reported annually. The main serotype isolated is O157:H7, and among non-O157 STEC, O145:NM is the most frequent strain. We studied the relationship of companion animals living in contact with a child with sporadic HUS, as carriers of Shiga toxin-producing Escherichia coli (STEC) strains. METHODOLOGY: Duplicate rectal swab samples were taken weekly from the household cat and dog at the home of a patient with HUS. Samples were plated on MacConkey and sorbitol MacConkey-CT agar. Confluent growth from each plate was screened for the presence of stx1, stx2 and rfbO157 gene by PCR assays. Up to 300 individual colonies taken from positive plates at screening were retested by PCR. RESULTS: The strain from the cat belonged to the highly virulent serotype O145:NM. Although this strain differed antigenically from the strain isolated from a child with HUS living in the same house, both carried the stx2, eae and ehxA virulence genes. The strain isolated from the dog belonged to the serotype O178:H19. CONCLUSIONS: An asymptomatic household cat may harbour the high virulent STEC strain, such as O145:NM, the second most frequently STEC serotype associated with HUS in Argentina. Companion animals are probably exposed to the same sources as the humans. More studies are needed to establish dogs and cats as sources of infection in the epidemiological cycle of infections caused by STEC strains, and to develop effective control strategies for this pathogen.

Antibody response to the epsilon toxin of Clostridium perfringens following vaccination of Lama glama crias.

BACKGROUND: Enterotoxaemia produced by Clostridium perfringens A, C and D is an important cause of mortality in young llamas. There is no data on antibody responses following vaccination with epsilon toxin. METHODOLOGY: Twenty-six L. glama crias were divided into four groups which were vaccinated with a commercial vaccine (Mancha Gangrena Enterotoxemia, Instituto Rosembusch Sociedad Anónima, Argentina) on days 0, 21 and 42 or left as unvaccinated controls. An indirect ELISA was compared with the mouse neutralization test (MNT) for measuring titers to C. perfringens type D epsilon toxin and used to determine titers in sera taken before vaccination and 16, 28, 49, 59, and 93 days later. RESULTS: The ELISA gave comparable results to the MNT and showed animals vaccinated once failed to develop raised titers. A week following a second vaccination, mean antibody titers rose significantly (P < 0.05) and 7/12 animals developed high titers which were present in only one animal at the end of the study (day 93). A third vaccination resulted in a decrease in mean antibody titers a week later. CONCLUSIONS: Llamas develop antibodies to Clostridium perfringens type D epsilon toxin after two vaccinations at a 21-day interval. Further studies are indicated to determine if these inoculations protect against enterotoxemia and the most appropriate vaccination schedule.