Whole-Genome Sequencing Analysis of Salmonella enterica Serovar Enteritidis Isolates in Chile Provides Insights into Possible Transmission between Gulls, Poultry, and Humans.

Salmonella enterica subsp. enterica serotype Enteritidis is a major cause of human salmonellosis worldwide; however, little is known about the genetic relationships between S Enteritidis clinical strains and S Enteritidis strains from other sources in Chile. We compared the whole genomes of 30 S Enteritidis strains isolated from gulls, domestic chicken eggs, and humans in Chile, to investigate their phylogenetic relationships and to establish their relatedness to international strains. Core genome multilocus sequence typing (cgMLST) analysis showed that only 246/4,065 shared loci differed among these Chilean strains, separating them into two clusters (I and II), with cluster II being further divided into five subclusters. One subcluster (subcluster 2) contained strains from all surveyed sources that differed at 1 to 18 loci (of 4,065 loci) with 1 to 18 single-nucleotide polymorphisms (SNPs), suggesting interspecies transmission of S Enteritidis in Chile. Moreover, clusters were formed by strains that were distant geographically, which could imply that gulls might be spreading the pathogen throughout the country. Our cgMLST analysis, using other S Enteritidis genomes available in the National Center for Biotechnology Information (NCBI) database, showed that S Enteritidis strains from Chile and the United States belonged to different lineages, which suggests that S Enteritidis regional markers might exist and could be used for trace-back investigations. IMPORTANCE: This study highlights the importance of gulls in the spread of Salmonella Enteritidis in Chile. We revealed a close genetic relationship between some human and gull S Enteritidis strains (with as few as 2 of 4,065 genes being different), and we also found that gull strains were present in clusters formed by strains isolated from other sources or distant locations. Together with previously published evidence, this suggests that gulls might be spreading this pathogen between different regions in Chile and that some of those strains have been transmitted to humans. Moreover, we discovered that Chilean S Enteritidis strains clustered separately from most of S Enteritidis strains isolated throughout the world (in the GenBank database) and thus it might be possible to distinguish the geographical origins of strains based on specific genomic features. This could be useful for trace-back investigations of foodborne illnesses throughout the world.

Serotype-associated polymorphisms in a partial rpoB gene sequence of Salmonella enterica.

Salmonella enterica is a zoonotic bacterium with more than 2500 serotypes, which affect a wide range of hosts and produce diverse clinical outcomes. Strain identification usually involves costly and time-demanding procedures. This paper describes the sequencing of a rpoB hypervariable gene segment (847 bp) that allows identification of serotypes in S. enterica strains isolated from several hosts. The nucleotide similarity values among S. enterica serotypes ranged from 98.23% to 99.88%, with potential usefulness for devising a simple one-step sequencing as a first approach for identification of S. enterica strains. In conclusion, the analysis of polymorphisms in the partial rpoB sequence can discriminate S. enterica strains at the subspecies level.

[Salmonella enterica: a review or the trilogy agent, host and environment and its importance in Chile]. / Salmonella enterica: una revisión de la trilogía agente, hospedero y ambiente, y su trascendencia en Chile.

Salmonella enterica is a major foodborne pathogen worldwide, being the main cause of outbreaks by food consumption in Chile. Despite all efforts deployed for control and prevention, the high incidence in people still persists, with several factors that could be influencing the epidemiological behavior of this infection. The objective of this review is to identify these factors belonging to the biological agent, the human host and the environment, which probably have a greater importance in Chile. Thus, priority areas for research of S. enterica are inferred, which hopefully will help to understand its spread in nature and its success as a wide host range pathogen. In the future, increased understanding of these determinants will facilitate the implementation of biosecurity and surveillance strategies for the prevention of disease in people and animals.

Quercetin Affects Erythropoiesis and Heart Mitochondrial Function in Mice.

Quercetin, a dietary flavonoid used as a food supplement, showed powerful antioxidant effects in different cellular models. However, recent in vitro and in vivo studies in mammals have suggested a prooxidant effect of quercetin and described an interaction with mitochondria causing an increase in O2 (∙-) production, a decrease in ATP levels, and impairment of respiratory chain in liver tissue. Therefore, because of its dual actions, we studied the effect of quercetin in vivo to analyze heart mitochondrial function and erythropoiesis. Mice were injected with 50 mg/kg of quercetin for 15 days. Treatment with quercetin decreased body weight, serum insulin, and ceruloplasmin levels as compared with untreated mice. Along with an impaired antioxidant capacity in plasma, quercetin-treated mice showed a significant delay on erythropoiesis progression. Heart mitochondrial function was also impaired displaying more protein oxidation and less activity for IV, respectively, than no-treated mice. In addition, a significant reduction in the protein expression levels of Mitofusin 2 and Voltage-Dependent Anion Carrier was observed. All these results suggest that quercetin affects erythropoiesis and mitochondrial function and then its potential use as a dietary supplement should be reexamined.

Genetic and phenotypic evidence of the Salmonella enterica serotype Enteritidis human-animal interface in Chile.

Salmonella enterica serotype Enteritidis is a worldwide zoonotic agent that has been recognized as a very important food-borne bacterial pathogen, mainly associated with consumption of poultry products. The aim of this work was to determine genotypic and phenotypic evidence of S. Enteritidis transmission among seabirds, poultry and humans in Chile. Genotyping was performed using PCR-based virulotyping, pulse-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Pathogenicity-associated phenotypes were determined with survival to free radicals, acidic pH, starvation, antimicrobial resistance, and survival within human dendritic cells. As result of PCR and PFGE assays, some isolates from the three hosts showed identical genotypic patterns, and through MLST it was determined that all of them belong to sequence type 11. Phenotypic assays show diversity of bacterial responses among isolates. When results were analyzed according to bacterial host, statistical differences were identified in starvation and dendritic cells survival assays. In addition, isolates from seabirds showed the highest rates of resistance to gentamycin, tetracycline, and ampicillin. Overall, the very close genetic and phenotypic traits shown by isolates from humans, poultry, and seabirds suggest the inter-species transmission of S. Enteritidis bacteria between hosts, likely through anthropogenic environmental contamination that determines infection of seabirds with bacteria that are potentially pathogenic for other susceptible organism, including humans.

Salmonella enterica: una revisión de la trilogía agente, hospedero y ambiente, y su trascendencia en Chile / Salmonella enterica: a review or the trilogy agent, host and environment and its importance in Chile

Salmonella enterica is a major foodborne pathogen worldwide, being the main cause of outbreaks by food consumption in Chile. Despite all efforts deployed for control and prevention, the high incidence in people still persists, with several factors that could be influencing the epidemiological behavior of this infection. The objective of this review is to identify these factors belonging to the biological agent, the human host and the environment, which probably have a greater importance in Chile. Thus, priority areas for research of S. enterica are inferred, which hopefully will help to understand its spread in nature and its success as a wide host range pathogen. In the future, increased understanding of these determinants will facilitate the implementation of biosecurity and surveillance strategies for the prevention of disease in people and animals.

Salmonella enterica es uno de los principales patógenos transmitidos por los alimentos en el mundo, siendo la primera causa de brotes de intoxicación alimentaria en Chile. A pesar de todos los esfuerzos de control y prevención desplegados, la incidencia en las personas se ha mantenido alta, por lo que diversos factores podrían estar influenciando el comportamiento epidemiológico de esta infección. El objetivo de esta revisión es describir factores referidos tanto al agente biológico, al hospedero humano y al medio ambiente, que podrían tener mayor trascendencia en Chile. De esta forma, se infieren ámbitos prioritarios para la investigación de S. enterica, que permitan entender su dispersión en la naturaleza y su éxito como patógeno de un amplio rango de hospederos. A futuro, el mayor conocimiento de estos determinantes facilitará la implementación de estrategias de bioseguridad y vigilancia para la prevención de la enfermedad en las personas y en los animales.

Identification of a gene cluster for the formation of extracellular polysaccharide precursors in the chemolithoautotroph Acidithiobacillus ferrooxidans.

A cluster of five genes, proposed to be involved in the formation of extracellular polysaccharide (EPS) precursors via the Leloir pathway, have been identified in the acidophilic autotroph Acidithiobacillus ferrooxidans. The order of the genes is luxA-galE-galK-pgm-galM, encoding a LuxA-like protein, UDP-glucose 4-epimerase, galactokinase, phosphoglucomutase, and galactose mutarotase, respectively. The gal cluster forms a single transcriptional unit and is therefore an operon. Two other putative genes of the Leloir pathway, galU, potentially encoding UDP-glucose pyrophosphorylase, and a gene designated galT-like, which may encode a galactose-1-phosphate uridylyltransferase-like activity, were found unlinked in the genome. Using semiquantitative reverse transcription-PCR, the genes of the gal operon were shown to be expressed more during growth in iron medium than in growth in sulfur medium. The functions of galE, pgm, galU, and the galT-like gene were validated by complementation of Escherichia coli mutants and by in vitro enzyme assays. The data suggest that A. ferrooxidans is capable of synthesizing the EPS precursors UDP-glucose and UDP-galactose. In addition, genes rfbA, -B, -C, and -D were identified in the genome of A. ferrooxidans, suggesting that it can also synthesize the EPS precursor dTDP-rhamnose. Since EPSs constitute the major bulk of biofilms, this study may provide an initial model for the metabolic pathways involved in biofilm formation in A. ferrooxidans and aid in understanding the role of biofilms in mineral leaching and the formation of acid mine drainage.