Identification et caractérisation de gènes impliqués dans la virulence de Salmonella typhi suite à une analyse globale par biopuces de l'infection de macrophages humains en culture

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Rapid emergence of multidrug resistant, H58-lineage Salmonella typhi in Blantyre, Malawi

INTRODUCTION: Between 1998 and 2010, S. Typhi was an uncommon cause of bloodstream infection (BSI) in Blantyre, Malawi and it was usually susceptible to first-line antimicrobial therapy. In 2011 an increase in a multidrug resistant (MDR) strain was detected through routine bacteriological surveillance conducted at Queen Elizabeth Central Hospital (QECH).

METHODS: Longitudinal trends in culture-confirmed Typhoid admissions at QECH were described between 1998-2014. A retrospective review of patient cases notes was conducted, focusing on clinical presentation, prevalence of HIV and case-fatality. Isolates of S. Typhi were sequenced and the phylogeny of Typhoid in Blantyre was reconstructed and placed in a global context.

RESULTS: Between 1998-2010, there were a mean of 14 microbiological diagnoses of Typhoid/year at QECH, of which 6.8% were MDR. This increased to 67 in 2011 and 782 in 2014 at which time 97% were MDR. The disease predominantly affected children and young adults (median age 11 [IQR 6-21] in 2014). The prevalence of HIV in adult patients was 16.7% [8/48], similar to that of the general population (17.8%). Overall, the case fatality rate was 2.5% (3/94). Complications included anaemia, myocarditis, pneumonia and intestinal perforation. 112 isolates were sequenced and the phylogeny demonstrated the introduction and clonal expansion of the H58 lineage of S. Typhi.

CONCLUSIONS: Since 2011, there has been a rapid increase in the incidence of multidrug resistant, H58-lineage Typhoid in Blantyre. This is one of a number of reports of the re-emergence of Typhoid in Southern and Eastern Africa. There is an urgent need to understand the reservoirs and transmission of disease and how to arrest this regional increase.

Ácidos orgânicos e probiótico na redução da colonização de Salmonella enterica Heidelberg em frangos de corte

In the food industry, Salmonella sp. and other pathogens represent a major threat as potential causes of foodborne diseases. The Salmonella enterica serovar Heidelberg (SH) has a great importance to public health, being currently the most frequent serotype in the official control analyzes carried out in Brazil’s Southern. In this region are concentrated the largest poultry flocks, contributing over 60 % of Brazilian production of chicken meat. To help prevent and minimize infections by pathogenic bacteria, in addition to attend the standards of biosecurity in the field and good hygiene and sanitizing programs in the industry, there is the possibility to control directly on the poultry breeding. Use of organic acids added in the diet of the fowls has an antibacterial action. In the other hand, the use of probiotics has been shown to prevent enteric diseases. This dissertation was divided into two chapters and aimed to evaluate the quantitative and qualitative reduction of SH in broiler chickens. In the first chapter the aim was to collect information on the prevalence rate of SH strain in broiler chickens. This coefficient was 100 %, being a suitable method for determining the frequency of salmonellosis in broiler chickens. In the second chapter five treatments were evaluated (a positive control and four tests), in order to reduce the colonization of SH in broilers, using organic acids and probiotic during production phase. In SH count related to the colonization of the cecum at 28 days of age differences were observed (p<0,05) among treatments. The Treatment 2 showed a count of 41 most probable number of bacterial cells per gram (MPN/g) with lower counts when compared to the positive control (5.106 MPN/g) and treatment 1 (1.354 MPN/g). The evaluation index of infected birds, there was a difference between treatments. In treatment 2, and treatment 4 reduced 58,34 % and 25 % respectively the number of fowls infected with SH. There was no effect of treatments on the performance. Administration of organic acids and probiotic used in the treatment 2 proved to be an effective tool in the SH control in broilers at 28 days of age.

Identificación de Enterobacter sakazakii y Salmonella enterica en preparados en polvo para lactantes suministrados en el Hospital de Maternidad "Dr. Raúl Argüello Escolán" de la red de salud pública de San Salvador, El Salvador

Los preparados en polvo para lactantes o PPL son sustancias utilizadas para la sustitución parcial o total de la alimentación materna; generalmente son utilizadas por los hospitales en niños prematuros, neonatos inmunodeficientes, hijos de madres VIH-positivas y/o con peso menor a los 2.5 Kg, según lo reporta la Organización Mundial para la Salud(FAO/OMS2007. Dentro de los peligros biológicos existentes en los PPL se encuentra E. sakazakii y Salmonella spp. En el entorno de la fabricación de las formúlas, cuando se trata de productos deshidratados como los PPL, no es posible utilizar la tecnología actual para producir PPL, que contenga bajas concentraciones de microrganismos, es decir, estos productos no pueden ser esterelizados. Para evitar posibles enfermedades o hasta la muerte es necesario la implementacion de buenas prácticas higiénicas (BPH) y buenas prácticas de manufactura (BPM. especialmente en paises como el nuestro.

Selection of polyvalent bacteriophages infecting Salmonella enterica serovar Choleraesuis

Background: Ideally, bacteriophages of pathogenic bacterial hosts should be polyvalent to be able to replicate in an alternative nonpathogenic bacterium. Thus, accidental infection by the original host can be avoided when bacteriophage lysates are used in biocontrol protocols. Results: From 15 wastewater samples, collected at different sites in the V Region in Chile, we selected three bacteriophages (FC, FP, and FQ) capable of productively infecting Salmonella enterica serovar Choleraesuis. By transmission electron microscopy (TEM) observation, the bacteriophages were found to belong to the order Caudoviridae. Molecular analyses indicated that FC, FP, and FQ contained double-stranded DNA genomes, of sizes similar to bacteriophage P22, and distinct recognition sites for the restriction endonucleases HaeIII and HindIII. Assays of host range revealed that the bacteriophages were polyvalent and thus capable of infecting different strains of Escherichia coli and other serovars of Salmonella . Conclusion: We have isolated newbacteriophages of the serovar Choleraesuiswith various potential applications in relation to this pathogenic bacterium.

Prevalence and characterization of Salmonella spp. in three shellfish-harvesting areas in France

Shellfish farming is an important economic activity in the Brittany and Normandy regions. However, a part of the production sites corresponds to relatively sensitive areas where the presence of faecal microorganisms is a major concern for shellfish and constitutes a possible health risk. Indeed, shellfish bioaccumulates in their tissues pathogenic contaminants present in water and can cause food-borne diseases such as salmonellosis. During a two-year study, we evaluated the presence of faecal indicators, measured the prevalence of Salmonella spp., isolated and characterized Salmonella spp. from three French shellfish-harvesting areas (shellfish and sediment) and their watersheds (from river water samples).

Diarrea crónica por Salmonella typhimurium en paciente inmunocompentente

La diarrea crónica de origen infeccioso en pacientes inmunocompetentes es un cuadro poco frecuente en países desarrollados, aunque ciertos patógenos, generalmente parásitos (Giardia lamblia, Isospora belli, Cryptosporidium, Cyclospora, Strongyloides, Ameba, Trichuris y Schistosoma) y algunas bacterias (Aeromonas, Plesiomonas, Campylobacter, Clostridium difficile, Salmonella o Mycobacterium tuberculosis) pueden ser causantes de diarrea persistente. Se presenta un caso de un paciente que presentó Salmonella typhimunium en el coprocultivo y se recuperó tras tratamiento con levofloxacino durante 7 días.

Eficácia de películas edíveis suplementadas com componentes de óleos essenciais no controlo de Listeria monocytogenes e Salmonella enterica Typhimurium

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014

Detecção de Salmonella spp. a partir de carcaças de bovinos obtidas durante o processamento em abatedouros-frigoríficos em Mato Grosso do Sul: resultados preliminares

2015

Detecção de Salmonella em tomates (Lycopersicon esculentum mill) pelos métodos fda-bam e mini vidas-biomérieux.

Resumo: Diversos surtos de Salmonella ocasionados pelo consumo de tomate contaminados com este micro-organismo têm sido relatados ultimamente, o que torna primordial a investigação sobre a presença desse patógeno nesse alimento. Métodos que permitam a avaliação rápida da presença de Salmonella em alimentos são de suma importância. O objetivo desse estudo foi comparar o método tradicional da Food and Drug Administration - Bacteriologycal Analytical Manual (FDA-BAM) com um método rápido da mini Vitek Immuno Diagnostic System Assay (Mini?Vidas-SLM)-bioMérieux, para detecção de Salmonella Brazil inoculada artificialmente na superfície de tomates. Foram analisadas 215 amostras de tomates inoculadas artificialmente com Salmonella Brazil com níveis de inóculos variando de 0,4 a 940 UFC/tomate. Os resultados obtidos mostraram que os métodos estudados apresentaram uma ótima concordância entre si, para todas as faixas de inóculo analisadas.

Salmonella enterica Serovar Typhimurium Exploits Inflammation to Modify Swine Intestinal Microbiota

Salmonella enterica serovar Typhimurium is an important zoonotic gastrointestinal pathogen responsible for foodborne disease worldwide. It is a successful enteric pathogen because it has developed virulence strategies allowing it to survive in a highly inflamed intestinal environment exploiting inflammation to overcome colonization resistance provided by intestinal microbiota. In this study, we used piglets featuring an intact microbiota, which naturally develop gastroenteritis, as model for salmonellosis. We compared the effects on the intestinal microbiota induced by a wild type and an attenuated S. Typhimurium in order to evaluate whether the modifications are correlated with the virulence of the strain. This study showed that Salmonella alters microbiota in a virulence-dependent manner. We found that the wild type S. Typhimurium induced inflammation and a reduction of specific protecting microbiota species (SCFA-producing bacteria) normally involved in providing a barrier against pathogens. Both these effects could contribute to impair colonization resistance, increasing the host susceptibility to wild type S. Typhimurium colonization. In contrast, the attenuated S. Typhimurium, which is characterized by a reduced ability to colonize the intestine, and by a very mild inflammatory response, was unable to successfully sustain competition with the microbiota.

Salmonella Typhimurium exploits inflammation to its own advantage in piglets

Salmonella Typhimurium (S. Typhimurium) is responsible for foodborne zoonotic infections that, in humans, induce self-limiting gastroenteritis. The aim of this study was to evaluate whether the wild-type strain S. Typhimurium (STM14028) is able to exploit inflammation fostering an active infection. Due to the similarity between human and porcine diseases induced by S. Typhimurium, we used piglets as a model for salmonellosis and gastrointestinal research. This study showed that STM14028 is able to efficiently colonize in vitro porcine mono-macrophages and intestinal columnar epithelial (IPEC-J2) cells, and that the colonization significantly increases with LPS pre-treatment. This increase was then reversed by inhibiting the LPS stimulation through LPS antagonist, confirming an active role of LPS stimulation in STM14028-intracellular colonization. Moreover, LPS in vivo treatment increased cytokines blood level and body temperature at 4 h post infection, which is consistent with an acute inflammatory stimulus, capable to influence the colonization of STM14028 in different organs and tissues. The present study proves for the first time that in acute enteric salmonellosis, S. Typhimurium exploits inflammation for its benefit in piglets.

Effect of cattle on Salmonella carriage, diversity and antimicrobial resistance in free-ranging wild boar (Sus scrofa) in northeastern Spain

Salmonella is distributed worldwide and is a pathogen of economic and public health importance. As a multi-host pathogen with a long environmental persistence, it is a suitable model for the study of wildlife-livestock interactions. In this work, we aim to explore the spill-over of Salmonella between free-ranging wild boar and livestock in a protected natural area in NE Spain and the presence of antimicrobial resistance. Salmonella prevalence, serotypes and diversity were compared between wild boars, sympatric cattle and wild boars from cattle-free areas. The effect of age, sex, cattle presence and cattle herd size on Salmonella probability of infection in wild boars was explored by means of Generalized Linear Models and a model selection based on the Akaike's Information Criterion. Prevalence was higher in wild boars co-habiting with cattle (35.67%, CI 95% 28.19-43.70) than in wild boar from cattle-free areas (17.54%, CI 95% 8.74-29.91). Probability of a wild boar being a Salmonella carrier increased with cattle herd size but decreased with the host age. Serotypes Meleagridis, Anatum and Othmarschen were isolated concurrently from cattle and sympatric wild boars. Apart from serotypes shared with cattle, wild boars appear to have their own serotypes, which are also found in wild boars from cattle-free areas (Enteritidis, Mikawasima, 4:b:- and 35:r:z35). Serotype richness (diversity) was higher in wild boars co-habiting with cattle, but evenness was not altered by the introduction of serotypes from cattle. The finding of a S. Mbandaka strain resistant to sulfamethoxazole, streptomycin and chloramphenicol and a S. Enteritidis strain resistant to ciprofloxacin and nalidixic acid in wild boars is cause for public health concern.