Adherence to and Invasion of Human Intestinal Cells by Arcobacter Species and Their Virulence Genotypes

The genus Arcobacter is composed of 17 species which have been isolated from various sources. Of particular interest are A. butzleri, A. cryaerophilus, and A. skirrowii, as these have been associated with human cases of diarrhea, the probable transmissionrroutes being through the ingestion of contaminated drinking water and food. To date, only limited studies of virulence traits in this genus have been undertaken. The present study used 60 Arcobacter strains isolated from different sources, representing 16 of the 17 species of the genus, to investigate their ability to adhere to and invade the human intestinal cell line Caco-2. In addition, the presence of five putative virulence genes (ciaB, cadF, cj1349, hecA, and irgA) was screened for in these strains by PCR. All Arcobacter species except A. bivalviorum and Arcobacter sp. strain W63 adhered to Caco-2 cells, and most species (10/16) were invasive. The most invasive species were A. skirrowii, A. cryaerophilus, A. butzleri, and A. defluvii. All invasive strains were positive for ciaB (encoding a putative invasion protein). Other putative virulence genes were present in other species, i.e., A. butzleri (cadF, cj1349, irgA, and hecA), A. trophiarum (cj1349), A. ellisii (cj1349), and A. defluvii (irgA). No virulence genes were detected in strains which showed little or no invasion of Caco-2 cells. These results indicate that many Arcobacter species are potential pathogens of humans and animals

SIDEROPHORES FOR SELECTIVE SOLID PHASE EXTRACTION OF STRATEGIC ELEMENTS

All over the world, industrial mining is leaving contaminated areas and dumps that, although being full of valuable metals, have high concentrations of toxic heavy metals that pollute the environment. The development of sustainable alternative biomining and bioremediation processes offers the potential to fully exploit these unexploited mining sites

Sanitary importance of arcobacter

The genus Arcobacter, belongs to the family Campylobacteraceae, along with. Some species, are considered emerging pathogens and have been associated with gastrointestinal diseases and bacteraemia in humans and with diarrhoea, mastitis and abortions in animals. Diarrhoea, is the most common clinical presentation in humans. However, still are not fully defined the routes of transmission, the virulence factors present in pathogenic strains have not been determined and the isolation and identification methods are still deficient. All of these factors contribute to a possible underestimation of the sanitary importance of this genus. In this thesis it has been demonstrated the existence of five new species of this genus. In their differentiation, new tools have been used, such as the Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI TOF) Multilocus Phylogenetic Analysis (MLPA) with concatenated 5 genes (rpoB, gyrB, hsp60, gyrA and atpA). Furthermore, the performance of 5 molecular identification methods was compared and none of them was useful for all the strains studied. In this regard, the updating of 16S rRNARFLP method for the identification of all species proved very useful. Moreover, in this thesis was a great diversity of species of this genus in samples of shellfish and sewage. Both matrices have epidemiological importance; however, they have been poorly studied. It was also demonstrated that the use in parallel of direct culturing and post enrichment, as well as the incubation under aerobic and microaerophilic, enhances the recovery and diversity of arcobacters. Regarding the virulence of Arcobacter spp., most of them proved to be potential enteropathogens for man, as they showed the presence of virulence factors and were able to adhere and invade human intestinal cells (Caco-2). Finally, it was demonstrated that Arcobacter may be confused with Campylobacter sp., generating an underestimation of their sanitary importance.El género Arcobacter incluye especies consideradas patógenos emergentes, ya que se han asociado con patologías gastrointestinales y bacteriemia en humanos y con diarrea, mastitis y abortos en animales. Sin embargo, aún no se ha definido completamente las rutas de transmisión, los factores de virulencia presentes en cepas patogénicas no se han determinado y los métodos de aislamiento e identificación aún son deficientes. Todo esto genera una subestimación de la importancia sanitaria de Arcobacter. En esta tesis doctoral se demostró la existencia de 5 nuevas especies de este género. Para su diferenciación se utilizó herramientas nuevas, tales como el Matrix Assisted Laser desorption Ionization Time of Flight (MALDI TOF) y un Multilocus Phylogenetic Analysis (MLPA) con 5 genes concatenados (rpoB, gyrB, hsp60, atpA y gyrA). Considerando el número de especies incluidas actualmente en el género (n=17), se evaluó si 5 de los métodos moleculares de identificación son todavía útiles para la identificación de las especies para las que se habían definido o si se generan confusiones, demostrrando que todos ellos generan algún error que osciló entre el 16,8% y 67,4% de las cepas estudiadas. En este sentido, la ampliación del método 16S rRNA-RFLP para poder identificar todas las especies resultó ser de gran utilidad. Por otra parte, en esta tesis se observó una gran diversidad de especies de este género en muestras de mariscos y aguas residuales. Ambas matrices han sido poco estudiados a pesar de su importancia epidemiológica. Más aún, se demostró que el uso en paralelo del cultivo por siembra directa y post enriquecimiento, además de incubación en aerobiosis y microaerofilia, favorece la recuperación de una mayor diversidad. Por otra parte, también se demostró que la mayoría de las especies de este género, en especial algunas cepas A. butzleri, A. cryaerophilus, A. skirrowii, A. trophiarum y A. defluvii, son potenciales enteropatógenos para el hombre, ya que presentaron factores de virulencia y fueron capaces de adherir e invadir células intestinales humanas (Caco-2). Por último, se demostró que Arcobacter puede ser confundido con Campylobacter sp., lo que puede contribuir aún más a subestimar su importancia sanitaria

Cronobacter condimenti sp. nov., isolated from spiced meat, and Cronobacter universalis sp. nov., a species designation for Cronobacter sp. genomospecies 1, recovered from a leg infection, water, and food ingredients

A re-evaluation of the taxonomic position of five strains, one assigned to Cronobacter sakazakii (strain 1330T), two previously assigned to Cronobacter genomospecies 1 (strains NCTC 9529T and 731) and two as Cronobacter turicensis (strains 96 and 1435) was carried out. The analysis included a phenotypic characterization, sequencing of the 16S rRNA gene and multilocus sequence analysis (MLSA) of seven housekeeping genes (atpD, fusA, glnS, gltB, gyrB, infB, ppsA; 3036 bp). The 16S rRNA gene sequence analysis and MLSA showed strain 1330T, isolated from spiced meat purchased in Slovakia, to form an independent phylogenetic line. Cronobacter dublinensis was the closest neighbour species on the basis of the MLSA. DNA–DNA reassociation and phenotypic analysis revealed that strain 1330T represented a novel species, for which the name Cronobacter condimenti sp. nov. is proposed, type strain 1330T = CECT 7863T, = LMG 26250T). The four bacterial strains NCTC 9529T, 731, 96 and 1435, isolated from water, a leg infectionand two food ingredients; onion powder and rye flour, repectively, showed on the phylogenetic tree to cluster together within an independent phylogenetic line, with Cronobacter turicensis as the closest species. The DNA–DNA hybridization data and the phenotypic characterization confirmed that these strains represented a novel species, for which the name Cronobacter universalis sp. nov. is proposed with type strain NCTC 9529T = CECT 7864T, = LMG 26249T

JC polyomavirus circulation in one-year surveillance in wastewater in Santiago, Chile

Human polyomavirus JC (JCPyV) is a widely distributed viral agent and because it high resistance against environmental conditions it is frequently recovered from diverse sources of water and is considered a good marker for human pollution. Phylogenetic analysis of JCPyV isolated in different part of the world has revealed 7 genotypes, which have been associated with specific populations or ethnics groups. This feature has been used to trace pre-historic and historic human migration patterns across the world. Although there are many reports describing genotypes distribution around the world, data on JCPyV genotypes in the southernmost areas of South America are scarce. The goal of this study is to detect and characterize the JCPyV that circulates in Santiago, Chile using sewage samples from wastewater treatment plants (WWTP). Sewage samples were obtained monthly during 1 year from three WWTPs which together process about 80% of wastewater generated in the city of Santiago, Chile. Our results show that JCPyV profusely circulates in Santiago, Chile, because it was detected in 80.56% of the samples, reinforcing the use of JCPyV as a feasible marker to assess human environmental pollution. JCPyV was detected in high frequency in influents and effluents samples, with the largest WWTPs showing the highest percentage of detection and viral loads. In the phylogenetic analysis the Chilean sequences clustered mainly with genotype 2A (Asian genotype). This is similar to that previously reported from Buenos Aires, Argentina and divergent to data from Brazil, where the circulation of European subtypes 1 and 4 and African subtypes 3 and 6 has been described

Performance of five molecular methods for monitoring Arcobacter spp

BACKGROUND: Bacteria belonging to the Arcobacter genus are emerging enteropathogens and potential zoonotic agents. Their taxonomy has evolved very rapidly, and there are presently 18 recorded species. The prevalence of species belonging to Arcobacter is underestimated because of the limitations of currently available methods for species identification. The aim of this study was to compare the performance of five PCR based methods that target regions of 16S rRNA, 23S rRNA or gyrA genes to identify Arcobacter species, and to review previous results reported in the literature using these methods. RESULTS: The five tested methods were found not to be reliable. They misidentified between 16.8% and 67.4% of the studied strains; this was dependent upon the target regions of the tested genes. The worst results obtained were for the identification of Arcobacter cryaerophilus and Arcobacter butzleri when the 23S rRNA gene was used as the target. These species were confused with many non-targeted species. CONCLUSION: Our results suggest that the known diversity of Arcobacter spp. in different environments could be expanded if reliable identification methods are applied in future studies

Genome Analysis of Planctomycetes Inhabiting Blades of the Red Alga

Porphyra is a macrophytic red alga of the Bangiales that is important ecologically and economically. We describe the genomes of three bacteria in the phylum Planctomycetes (designated P1, P2 and P3) that were isolated from blades of Porphyra umbilicalis (P.um.1). These three Operational Taxonomic Units (OTUs) belong to distinct genera; P2 belongs to the genus Rhodopirellula, while P1 and P3 represent undescribed genera within the Planctomycetes. Comparative analyses of the P1, P2 and P3 genomes show large expansions of distinct gene families, which can be widespread throughout the Planctomycetes (e.g., protein kinases, sensors/response regulators) and may relate to specific habitat (e.g., sulfatase gene expansions in marine Planctomycetes) or phylogenetic position. Notably, there are major differences among the Planctomycetes in the numbers and sub-functional diversity of enzymes (e.g., sulfatases, glycoside hydrolases, polysaccharide lyases) that allow these bacteria to access a range of sulfated polysaccharides in macroalgal cell walls. These differences suggest that the microbes have varied capacities for feeding on fixed carbon in the cell walls of P.um.1 and other macrophytic algae, although the activities among the various bacteria might be functionally complementary in situ. Additionally, phylogenetic analyses indicate augmentation of gene functions through expansions arising from gene duplications and horizontal gene transfers; examples include genes involved in cell wall degradation (e.g., κ-carrageenase, alginate lyase, fucosidase) and stress responses (e.g., efflux pump, amino acid transporter). Finally P1 and P2 contain various genes encoding selenoproteins, many of which are enzymes that ameliorate the impact of environmental stresses that occur in the intertidal habitat

Presencia de Violencia en Relaciones Románticas en Adolescentes de la Ciudad de Río Bueno, Sur de Chile

The objective of the present research is to portray violence in teen students’ romantic relationships in a high school in the city of Rio Bueno, XIV Region. The 239 students who participated were measured by the “Escala de Maltrato en la Pareja Forma A” (Rey, 2009) which was adapted by the DIULA Nº025/201 Research Project Team. In order to apply this research tool, participants were required to have been in a relationship in the last twelve months. Main results show that 90.8% of the participants stated to have been victim of some kind of violence at least once. On the other hand, 91.2% affirmed to have been a victimizer. There wasn’t any significant difference in regards to gender.La presente investigación tuvo por objetivo caracterizar situaciones de violencia que se presentan en las relaciones románticas de estudiantes adolescentes de Río Bueno, ciudad ubicada en la décima cuarta región de Chile. Los participantes fueron 239 estudiantes a quienes se aplicó la Escala de Maltrato en la Pareja Forma A (Rey 2009), adaptado por el equipo del Proyecto de Investigación DIULA N°025/2011. Para poder participar en el estudio los/las estudiantes debían estar o haber estado en una relación afectiva en los últimos doce meses. Los principales resultados obtenidos muestran que un 90,8% de los participantes declaró haber recibido al menos en una ocasión algún tipo de violencia, y que en cuanto a violencia ejercida un 91,2% de los participantes señaló ejercer al menos algún tipo de violencia, en ambos casos sin diferencia significativa entre géneros