The aim of this study was to investigate the role and clinical course of verotoxigenic Escherichia coli (VTEC) infections in children with acute diarrhoea from Argentina, the country with the highest worldwide incidence of haemolytic uraemic syndrome (HUS). To accomplish this objective, 437 samples from children up to 6 years old with acute diarrhoea were collected and processed. More than 60 % of the children studied presented watery or mucous diarrhoea without blood, and in 25.2 % of the cases the samples contained blood. In a first screening, a multiplex PCR was performed to detect the presence of the vt 1 , vt 2 , eae, ehxA and saa virulence genes. The strains were then isolated and analysed to characterize their serotypes, virulence genes, antibiotic susceptibility profiles and verotoxin (VT) production. Forty-four of the 437 samples (10.1 %) were positive for VTEC virulence genes. VTEC-infected patients presented different types of diarrhoea (27.3 % belonged to the non-bloody type). Several serotypes and virulence genotypes were found. Isolates belonged to the serotypes O157 : H7, O145 : H " , O26 : H11, O121 : H19, O111 : H2 and O118 : H2. HUS developed in 16 (36.4 %) patients positive for VTEC virulence genes. All of the VTEC isolates produced a cytopathic effect on Vero cell monolayers, confirming the ability to express VT. Despite most strains being sensitive to all of the antimicrobials studied, a positive association between clinical progression to HUS and antibiotic therapy was observed for the total number of patients studied, as well as for the VTEC + group. In conclusion, the data obtained in this study increase our knowledge of the role and clinical course of VTEC infection in childhood acute diarrhoea beyond bloody diarrhoea, and might be considered for the prevention, diagnosis and management of this disease. It is possible that the optimal approach for VTEC diagnosis could be using multiplex PCR to search for the presence of the vt 1 , vt 2 , eae and ehxA genes.

In order to study the seasonality of haemolytic uraemic syndrome (HUS) and verotoxigenic Escherichia coli (VTEC) infection in children, 437 patients under 6 years of age with acute diarrhoea were studied, 8% of whom progressed to HUS. VTEC was found in 10% of all of the stool samples analysed and seasonal occurrence of HUS (p < 0.01) was confirmed. VTEC infection was more prevalent in warm months, although the differences were not statistically significant. Moreover, a significant difference in the detection of O157:H7 serotype and in the vt profile between cold and warm months (autumn and winter; spring and summer, respectively) was established. The O157:H7 serotype was isolated more frequently during warm months. Moreover, a predominance of vt (2) was noted, which was partially replaced by the combination of vt (1) with vt (2) in the cold season. The results of this study indicate the seasonal variation of the disease and the presence of serotype O157:H7 and the vt types. They also reinforce the need to develop prevention programmes considering the seasonal pattern of the disease, which would generate an impact on public health. Control strategies of the pathogen in cattle in the most risky season of the year would also be of benefit.

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Verotoxigenic Escherichia coli (VTEC) are one of the most important emerging foodborne pathogens and the principal cause of hemolytic uremic syndrome (HUS). This entity has been recognized worldwide as a priority issue in the field of zoonoses and public health, and Argentina is the country with the highest incidence of HUS in children less than 5 years of age.The lack of specific treatment, combined with the high morbidity rate of VTEC infection, makes prevention the main tool for reducing the incidence of HUS. The current work aimed at assessing the factors associated with sporadic VTEC infection in children with acute diarrhea from the Central Eastern area of Argentina where the incidence rate of HUS in children under 5 is the highest worldwide. A univariate analysis was performed to identify potential factors associated with VTEC infection by calculating odds ratios (OR) with 95% confidence intervals (CI). Then, a multivariate logistic regression model was performed. Interaction and association between significant factors were checked. "Recent consumption of food prepared outside home" (OR: 2.4, 95% CI 1.05-5.7) and "recent vegetables consumption" (OR=0.4; 0.2-0.8) were identified as independent factors associated with VTEC infection. We believe that the data obtained from this study further the current knowledge about the epidemiology of VTEC infection in Argentina and could be considered when planning strategies for the prevention of the disease.

Control of ovine brucellosis with subcellular vaccines can solve some drawbacks associated with the use of Brucella melitensis Rev.1. Previous studies have demonstrated that the polymeric antigen BLSOmp31 administered by parenteral route was immunogenic and conferred significant protection against B. ovis in rams. Immunization with BLSOmp31 by conjunctival route could be efficient for the induction of mucosal and systemic immune responses. In this work, we evaluated the conjunctival immunization using a thermoresponsive and mucoadhesive in situ gel composed of Poloxamer 407 (P407) and chitosan (Ch) as vaccine delivery system for BLSOmp31 in rams. Serum samples, saliva, lacrimal, preputial and nasal secretions were analyzed to measure specific IgG and IgA antibodies. Cellular immune response was evaluated in vivo and in vitro. Immunization with BLSOmp31-P407-Ch induced high IgG antibody levels in serum and preputial secretions which remained at similar levels until the end of the experiment. Levels of IgG in saliva, lacrimal and nasal secretions were also higher compared to unvaccinated control group but decreased more rapidly. IgA antibodies were only detected in nasal and preputial secretions. BLSOmp31-P407-Ch stimulated a significant cellular immune response in vivo and in vitro. The induction of systemic and local immune responses indicates a promising potential of P407-Ch for the delivery of BLSOmp31 by conjunctival route.

Leptospirosis is an infectious disease caused by pathogenic Leptospira that affect humans and animals. This disease is complex and non-eradicable in nature. Therefore, the understanding of it is epidemiology in different environments is crucial to implement prevention and control measures. The prevalence of Leptospira infection in beef cattle farms is affected by multiple environmental, management and individual factors. In this study, a cross-sectional serological survey was carried on to estimate the prevalence of Leptospira antibodies in beef cattle in Tandil and Ayacucho Departments (Buenos Aires Province) and to identify risk factors and spatial clusters associated with seropositivity. Using a probabilistic two-stage sampling, 25 farms and 15 animals per farm were selected. The Microagglutination Test was used to analize all serum samples. Bivariate and multivariate analyses were performed. Seventy-three out of 375 cows were seropositive, representing a positivity rate of 19.47% (95% CI: 10.51–28.42), with Sejroe and Pomona being the most reactive serogroups: 9.33% (95% CI: 6.26–12.41) and 8.27% (95% CI: 5.35–11.19), respectively. The prevalence in Ayacucho was 23.11% (95% CI: 10.05–36.17), and in Tandil, 14% (95% CI: 3.25–24.75). The animals from Ayacucho presented 2.01 (1.16–3.49) more chances of being positive compared with those from Tandil (p &lt; 0.01). After the Generalized Linear Mixed Model (GLMM) with random effect of farm-level risk, the presence of lagoons (OR: 7.32, 95% CI: 1.68–31.8, p &lt; 0.05) and undulating terrain (OR: 0.24, 95% CI: 0.07–0.74, p &lt; 0.05) were associated with bovine leptospirosis. Four spatial clusters with higher rates of seropositivity were detected. A new GLMM was performed with the significant variables detected in the first GLMM and a new variable, “being inside the spatial cluster,” being the only one that remained significant (OR: 9.58, 95% CI: 3.39–27.08, p &lt; 0.0001). The animals inside the clusters belonged to farms with a greater presence of creeks (OR: 9.03, 95% CI: 3.37–24.18, p &lt; 0.0001), higher accumulated rainfall (OR: 1.01, 95% CI: 1–1.01, p &lt; 0.0001) and less undulating terrain (OR: 0.18, 95% CI: 0.10–0.35, p &lt; 0.0001). We conclude that Leptospira is seroprevalent in beef cattle in Tandil and Ayacucho Departments, especially in the latter, where the largest cattle farms are located. Prevalence of seropositivity animals was associated with selected environmental risk factors.

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Trichinellosis is an emergent zoonosis in several regions of the world and it is considered a public health problem. Trichinellosis represent one of the most important zoonotic diseases in Argentina. The purpose of this study was to develop a real-time PCR assay with internal control to detect Trichinella spiralis DNA in samples of swine muscles and its derivatives. PCR amplification of DNA from muscle samples was performed by real-time PCR with an internal porcine DNA amplification control. The developed PCR assay specifically detects T. spiralis showing no amplification with other Trichinella genotypes and showed an estimated sensitivity of 0.024 larvae per gram in swine muscle samples. From the 21 samples analyzed, five samples negative by enzymatic digestion were positive by real-time PCR, which demonstrates that this technique is capable of detecting T. spiralis in cases when enzymatic digestion cannot. In this work, a new molecular assay with internal control for T. spiralis detection was successfully developed. PRACTICAL APPLICATIONSTrichinellosis is considered a public health problem in Argentina and also represents an economic problem in porcine animal production and food safety. Due to the predominantly zoonotic importance of infection, the main efforts have focused on the control of Trichinella or the elimination of Trichinella from the food chain. In our country, most human infections are caused by Trichinella spiralis, which is transmitted mainly by the ingestion of raw or undercooked infected swine meat or its derivative products. New controls in order to improve food safety for consumers should be developed. This new real-time PCR assay with internal control shows a substantial increase in diagnostic sensitivity in comparison with muscle artificial digestion and the possibility to avoid false negative results. This specific PCR for T. spiralis may be useful for detection of infection at early stages in humans and food animals.