Vaginal Microbiome Characterization of Nellore Cattle Using Metagenomic Analysis

Understanding of microbial communities inhabiting cattle vaginal tract may lead to a better comprehension of bovine physiology and reproductive health being of great economic interest. Up to date, studies involving cattle microbiota are focused on the gastrointestinal tract, and little is known about the vaginal microbiota. This study aimed to investigate the vaginal microbiome in Nellore cattle, heifers and cows, pregnant and non-pregnant, using a culture independent approach. The main bacterial phyla found were Firmicutes (~40–50%), Bacteroidetes (~15–25%) and Proteobacteria (~5–25%), in addition to ~10–20% of non-classified bacteria. 45–55% of the samples were represented by only ten OTUs: Aeribacillus, Bacteroides, Clostridium, Ruminococcus, Rikenella, Alistipes, Bacillus, Eubacterium, Prevotella and non-classified bacteria. Interestingly, microbiota from all 20 animals could be grouped according to the respiratory metabolism of the main OTUs found, creating three groups of vaginal microbiota in cattle. Archaeal samples were dominated by the Methanobrevibacter genus (Euryarchaeota, ~55–70%). Ascomycota was the main fungal phylum (~80–95%) and Mycosphaerella the most abundant genus (~70–85%). Hormonal influence was not clear, but a tendency for the reduction of bacterial and increase of archaeal populations in pregnant animals was observed. Eukaryotes did not vary significantly between pregnant and non-pregnant animals, but tended to be more abundant on cows than on heifers. The present work describes a great microbial variability in the vaginal community among the evaluated animals and groups (heifers and cows, pregnant and non-pregnant), which is significantly different from the findings previously reported using culture dependent methods, pointing out the need for further studies on this issue. The microbiome found also indicates that the vaginal colonization appears to be influenced by the gastrointestinal community.     

Oral Outbreak of Chagas Disease in Santa Catarina,Brazil: Experimental Evaluation of a Patient’s Strain

Chagas disease is a worldwide public health problem. Although the vectorial transmission of Chagas disease has been controlled in Brazil there are other ways of transmission, such as the ingestion of T. cruzi contaminated food, which ensures the continuation of this zoonosis. Here, we demonstrate the influence of the inoculation route on the establishment and development of the SC2005 T. cruzi strain infection in mice. Groups of Swiss mice were infected intragastrically (IG) or intraperitoneally (IP) with the T. cruzi SC2005 strain derived from an outbreak of oral Chagas disease. The results revealed that 100% of IP infected mice showed parasitemia, while just 36% of IG infected showed the presence of the parasite in blood. The parasitemia peaks were later and less intense in the IG infected mice. Mortality of the IP infected animals was more intense and earlier when compared to the IG infected mice. In the IP infected mice leucopenia occurred in the early infection followed by leucocytosis, correlating positively with the increase of the parasites. However, in the IG infected mice only an increase in monocytes was observed, which was positively correlated with the increase of the parasites. Histopathological analyses revealed a myotropic pattern of the SC2005 strain with the presence of inflammatory infiltrates and parasites in different organs of the animals infected by both routes as well as fibrosis foci and collagen redistribution. The flow cytometric analysis demonstrated a fluctuation of the T lymphocyte population in the blood, spleen and mesenteric lymph nodes of the infected animals. T. cruzi DNA associated with the presence of inflammatory infiltrates was detected by PCR in the esophagus, stomach and intestine of all infected mice. These findings are important for the understanding of the pathogenesis of T. cruzi infection by both inoculation routes.     

High ultraviolet C resistance of marine Planctomycetes

Planctomycetes are bacteria with particular characteristics such as internal membrane systems encompassing intracellular compartments, proteinaceous cell walls, cell division by yeast-like budding and large genomes. These bacteria inhabit a wide range of habitats, including marine ecosystems, in which ultra-violet radiation has a potential harmful impact in living organisms. To evaluate the effect of ultra-violet C on the genome of several marine strains of Planctomycetes, we developed an easy and fast DNA diffusion assay in which the cell wall was degraded with papain, the wall-free cells were embedded in an agarose microgel and lysed. The presence of double strand breaks and unwinding by single strand breaks allow DNA diffusion, which is visible as a halo upon DNA staining. The number of cells presenting DNA diffusion correlated with the dose of ultra-violet C or hydrogen peroxide. From DNA damage and viability experiments, we found evidence indicating that some strains of Planctomycetes are significantly resistant to ultra-violet C radiation, showing lower sensitivity than the known resistant Arthrobacter sp. The more resistant strains were those phylogenetically closer to Rhodopirellula baltica, suggesting that these species are adapted to habitats under the influence of ultra-violet radiation. Our results provide evidence indicating that the mechanism of resistance involves DNA damage repair and/or other DNA ultra-violet C-protective mechanism.     

Kinetics of growth and ethanol formation from a mix of glucose/xylose substrate by Kluyveromyces marxianus UFV-3

The fermentation of both glucose and xylose is important to maximize ethanol yield from renewable biomass feedstocks. In this article, we analyze growth, sugar consumption, and ethanol formation by the yeast Kluyveromyces marxianus UFV-3 using various glucose and xylose concentrations and also under conditions of reduced respiratory activity. In almost all the conditions analyzed, glucose repressed xylose assimilation and xylose consumption began after glucose had been exhausted. A remarkable difference was observed when mixtures of 5 g L^?1 glucose/20 g L^?1 xylose and 20 g L^?1 glucose/20 g L^?1 xylose were used. In the former, the xylose consumption began immediately after the glucose depletion. Indeed, there was no striking diauxic phase, as observed in the latter condition, in which there was an interval of 30 h between glucose depletion and the beginning of xylose consumption. Ethanol production was always higher in a mixture of glucose and xylose than in glucose alone. The highest ethanol concentration (8.65 g L^?1) and cell mass concentration (4.42 g L^?1) were achieved after 8 and 74 h, respectively, in a mixture of 20 g L^?1 glucose/20 g L^?1 xylose. When inhibitors of respiration were added to the medium, glucose repression of xylose consumption was alleviated completely and K. marxianus was able to consume xylose and glucose simultaneously.     

Analytical and numerical analysis of human dental occlusal contact

The knowledge of contact forces in teeth surfaces during mastication or para-functional movements can help to understand processes related to friction and wear of human dental enamel. The development of a numerical model for analysis of the occlusal contact between two antagonistic teeth is proposed, which includes three basic steps: the characterisation of the surface roughness, its homogenisation using an assumed distribution function and the numerical determination of the resulting forces. Finite element strain results for the main different asperities are statistically combined, deriving the predicted macroscopic behaviour of the interface. Axisymmetric and 3D numerical models with an elasto-plastic constitutive law are used to simulate micro-indentations and micro-contacts, respectively. The contact is allowed to occur locally in planes not necessarily parallel to the surface's mean plane, a problem for which there is no analytical solution. The three identified parameters, homogenised surface hardness (3.68 GPa), surface yield stress (3.08 GPa) and static friction coefficient (0.23), agree with the experimental values reported in the literature.     

Cross-Sectional Relationship Between Chronic Stress and Mineral Concentrations in Hair of Elementary School Girls

Chronic stress exposure is associated with diverse negative health outcomes. It has been hypothesised that stress may also negatively affect the body's mineral status. This study investigates the association between chronic stress and long-term mineral concentrations of calcium (Ca), copper (Cu), iron (Fe), magnesium (Mg), phosphorus (P) and zinc (Zn) in scalp hair among elementary school girls. Complete information on child-reported stress estimates (Coddington Life Events Scale (CLES)), hair cortisone and hair mineral concentrations, and predefined confounders in the stress–mineral relationship (i.e. age, body mass index, physical activity, diet, hair colour and parental education) was provided cross-sectionally for 140 girls (5–10 years old). The relationship between childhood stress measures (predictor) and hair minerals (outcome) was studied using linear regression analysis, adjusted for the abovementioned confounders. Hair cortisone concentrations were inversely associated with hair mineral concentrations of Ca, Mg, Zn and the Ca/P ratio. Children at risk by life events (CLES) presented an elevated Ca/Mg ratio. These findings were persistent after adjustment for confounders. This study demonstrated an independent association between chronic stress measures and hair mineral levels in young girls, indicating the importance of physiological stress–mineral pathways independently from individual or behavioural factors. Findings need to be confirmed in a more heterogeneous population and on longitudinal basis. The precise mechanisms by which stress alters hair mineral levels should be further elucidated.     

Pleural Effusion an Unusual Feature of Paracoccidioidomycosis: Report of Two New Cases with a Systematic Review of the Literature

Paracoccidioidomycosis (PCM) is an endemic disease restricted geographically to Latin America. Brazil accounts for about 80 % of the reported cases, and lungs are most frequently affected. A suggestive radiograph of PCM may only be seen late in the course of the disease. At the beginning, it mimics tuberculosis. On the other hand, pleural effusion on rare occasions has been reported in PCM. For this reason, we report two cases with such uncommon manifestation. Our first patient presented pleural effusion probably caused by PCM (a previously unreported cause of lung mass accompanied by effusion); the second with proved pleural effusion due to PCM. A systematic review of the literature was done.     

Breast cancer tissue slices as a model for evaluation of response to rapamycin

Rapamycin is a selective inhibitor of the mammalian target of rapamycin (mTOR), a regulator kinase that integrates growth factors signaling via the phosphoinositide-3-kinase pathway and that has emerged as a novel therapeutic modality in breast cancer (BC). We propose a pre-clinical “ex-vivo” personalized organotypic culture of BC that preserves the microenvironment to evaluate rapamycin-mediated gene expression changes. Freshly excised ductal invasive BC slices, 400 μm thick (n=30), were cultured in the presence or absence (control) of rapamycin (20 nM) for 24 h. Some slices were formalin-fixed for immunohistochemical determinations and some were processed for microarray analysis. Control slices in culture retained their tissue morphology and tissue viability (detected by BrdU uptake). The percentage of proliferating cells (assessed by Ki67) did not change up to 24 h of treatment. Immunohistochemical evaluation of p-AKT, p-mTOR, p-4EBP1 and p-S6K1 indicated that AKT/mTOR pathway activation was maintained during cultivation. For microarray analysis, slices were divided into two groups, according to the presence/absence of epidermal growth factor receptor-type 2 and analyzed separately. Limited overlap was seen among differentially expressed genes after treatment (P<0.01) in both groups suggesting different responses to rapamycin between these BC subtypes. Ontology analysis indicated that genes involved in biosynthetic processes were commonly reduced by rapamycin. Our network analysis suggested that concerted expression of these genes might distinguish controls from treated slices. Thus, breast carcinoma slices constitute a suitable physiological tool to evaluate the short-term effects of rapamycin on the gene profile of individual BC samples.     

Effect of temperature on the development time and life-time fecundity of Trichopria anastrephae parasitizing Drosophila suzukii

The aim of this study was to evaluate the effect of temperature (10, 15, 20, 25, 30 and 35°C), on the development time and life-time fecundity of Trichopria anastrephae Lima, 1940 (Hymenoptera: Diapriidae) parasitizing Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae). D. suzukii pupae that were up to 24-hr old were submitted to T. anastrephae parasitism for 24 hr. They were placed in plastic containers (50 ml) (ten pupae per container) in climatic chambers at temperatures of 10, 15, 20, 25, 30 and 35°C ± 1°C with a relative humidity of 70% ± 10% and a 12 hr photoperiod. For the adult phase, T. anastrephae couples that were up to 24 hr old were each placed in plastic cages (300 ml) and kept at the same temperatures cited above until their deaths. The higher numbers of parasitism and offspring production were obtained at temperatures between 15 and 25ºC. At the temperatures of 10 and 35ºC, there was no emergence of individuals. The lower thermal threshold (Tt) for the egg to adult period was ≈ 14.6°C for males and females with thermal constants (K) of 384.61 and 432.90, respectively. In terms of the fertility life table, T. anastrephae at 20 and 25°C presented shorter generation time (T) and higher net reproductive rates (Ro) in relation to other temperatures. The data show the ability of T. anastrephae to adapt to different thermal conditions, which is important for biological control programmes of D. suzukii.