Ativação local da rota da quinurenina por moduladores inflamatórios durante a meningite bacteriana.

Activation of the kynurenine (KYN) pathway (KP) by modulators of immune system has been observed during several neurological diseases. Here we assessed the association of chemo-/cytokine levels with the concentration of KP metabolites in cerebrospinal fluid (CSF) and plasma samples from patients with bacterial meningitis (BM). All samples were collected from 42 patients diagnosed with acute bacterial meningitis (ABM), aseptic meningitis, tuberculous meningitis and patients without infection neurological disorders. CSF and plasma concentration of metabolites from the KP was assessed by high pressure liquid chromatography (HPLC) and cytokines and chemokines by Bio-plex 200 suspension array system. Concentrations of the KP metabolites KYN and kynurenic acid (KYNA) were significantly higher in CSF of patients with ABM compared to other groups. Tryptophan (TRP), anthranilic acid (AA), 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) did not show statistical significance, although some of them presented a good accumulation during ABM. The expression of TNF-alpha, IL-6, IL-1beta, IFN-gamma, IL-10, IL-1 receptor antagonist (IL-1Ra), MIP-1alpha, MIP-1beta, MCP-1 and G-CSF was about 100-fold higher in CSF from ABM patients than other infected groups. In all CSF and plasma samples, the concentration of IL-2, IL-12(p70), IL-4, IL-8 and GM-CSF was not significant. ABM still showed significant concentrations of IL-6, IL-10, IL-1Ra and MCP-1 in plasma samples. Based on the comparison of KP metabolites concentrations between plasma and CSF samples we conclude that the activation of the tryptophan pathway upon BM occurs within the brain. This increase in KP metabolites is most due to activation of the KP by molecules as IFN-gamma and TNF-alpha in response to infection.

Caracterização da resistência do algodoeiro a Ramularia areola e variabilidade molecular do patógeno

This research was conducted with the aim to study the genetic and pathogenic structure of Ramularia areola isolates collected in Brazil and to characterize the resistance response in cotton plants to ramularia spot. The genetic variability of 28 isolates of R. areola was studied using RAPD markers. The pathogenicity evaluation was realized by the inoculation of 6 isolates on cotton varieties Guazuncho-2 (Gossypium hirsutum) and VH8-4602 (Gossypium barbadense). The inheritance of disease resistance was studied using an artificially inoculated population of F2 individuals derived from the intercross of Guazuncho-2 (susceptible variety) end VH8-4602 (resistant variety), and also the parents and F1 individuals. Molecular polymorphism between the G. hisutum varieties DeltaOpal (suscetible) and CNPA CO-11612 (resistant) was estimated by 118 SSR and 24 AFLP markers. The parental genotypes Guazuncho-2 and VH8-4602 were selected for mapping, and then Recombinant Inbred Lines (RIL´s) derived from this crossing were evaluated with SSR 12 markers. The analysis of population structure of R. areola revealed that the three subpopulations were genetically simillar (Gst=0.18), and the isolates from Goiás and Minas Gerais were more similar to each other (0,92). This probability can be related to the relatively high gene flow among the three subpopulations (Nm=2.20). The isolates R. areola 9.1, from Minas Gerais State and 8.1 and 8.3 from Goiás State were the most aggressive ones to the susceptible variety Guazuncho-2. The variety VH8-4602 presented high level of resistance to ramularia spot. No differential interaction was observed between the pathogens and the analyzed varieties, and the resistance was classified as horizontal. The quantification of disease by number of necrotic lesions and number of spores in individual plants of F1 and F2 generations from the crossing between the varieties Guazuncho-2 and VH8-4602 presented continuous distribution, suggesting polygenic resistance. The resistance is probabilly recessive, since necrotic lesions and sporulation were observed on F1 plants. The molecular polymorphism between DeltaOpal e CNPA CO-11612 lineages was low (6%), then would be difficult to accomplish molecular mapping of disease resistance using this intercross. With the genotyping of the RIL s it was verified that 25% of the markers segregated in the proportions proposed by Mendel s Law and 75% of the studied markers presented segregation distortion in favor to the parental G. hirsutum. Both the low genetic variability of the pathogen and the number of resistance genes suggest that durable genetic resitance may be achieved

Prevalência e susceptibilidade antimicrobiana de Staphylococcusspp. em quadros de saúde e doença periodontal

The aim of this study was determine the prevalence and antimicrobial susceptibility of Staphylococcus spp. from patients with periodontal disease and periodontally healthy, correlate them with factors to host, local environment and traits of the diseases. To this, thirty adults from 19 to 55 years old were selected. They had not periodontal treatment and no antibiotic or antimicrobial was administered during three previous months. From these individuals, sites periodontally healthy, with chronic gingivitis and/or periodontitis were analyzed. Eighteen subgingival dental biofilm samples were collected through sterile paper points being six from each tooth randomly selected, representing conditions mentioned. They were transported to Oral Microbiology laboratory, plated onto Mannitol Salt Agar (MSA) and incubated at 370C in air for 48 h. Staphylococcus spp. were identified by colonial morphology, Gram stain, catalase reaction, susceptibility to bacitracin and coagulase activity. After identification, strains were submitted to the antibiotic susceptibility test with 12 antimicrobials, based on Kirby-Bauer technique. To establish the relation between coagulase-negative Staphylococcus (CSN) presence and their infection levels and host factors, local environment and traits of diseases were used Chi-square, Mann-Whitney and Kruskal-Wallis tests to a confidence level of 95%. 86,7% subjects harbored CSN in 11,7% periodontal sites. These prevalence were 12,1% in healthy sites, 11,7% in chronic gingivitis, 13,5% in slight chronic periodontitis, 6,75% in moderate chronic periodontitis and in sites with advance chronic periodontitis was not isolated CSN, without difference among them (p = 0,672). There was no significant difference to presence and infection levels of CSN as related to host factors, local environm ent and traits of the diseases. Amongst the 74 samples of CSN isolated, the biggest resistance was observed to penicillin (55,4%), erythromycin (32,4%), tetracycline (12,16%) and clindamycin (9,4%). 5,3% of the isolates were resistant to oxacilin and methicillin. No resistance was observed to ciprofloxacin, rifampicin and vancomycin. It was concluded that staphylococci are found in low numbers in healthy or sick periodontal sites in a similar ratio. However, a trend was observed to a reduction in staphylococci occurrence toward more advanced stages of the disease. This low prevalence was not related to any variables analyzed. Susceptibility profile to antibiotics demonstrates a raised resistance to penicillin and a low one to methicillin. To erythromycin, tetracycline and clindamycin was observed a significant resistance

Marcadores fenotípicos para caracterização de caprinos com diferentes níveis de resistência às endoparasitoses gastrintestinais

The aim of this study was to evaluate and characterize phenotypically goats with different levels of resistance to gastrointestinal nematodes. For a period of 93 days, 60 F2 goats originated from ½ Saanen and ½ Anglo- nubian animals were kept in the same area of pasture. Every seven days, feces and blood were collected for eggs per gram counts of feces (EPG) and cultures of feces and to determinate the number of eosinophils, packed cell volume and total plasma protein, respectively. On the same day, the animals were weighed and submitted to body score condition and FAMACHA method to worm control. Based on the average of EPG, the twelve animals with the highest average (susceptible group) and the twelve animals with the lowest average of EPG (resistant group) were selected, slaughtered and necropsied to recovery, counting andparasites identification. The resistant animals present lower EPG mean (P <0.0001) and 4.7 folder less parasites than susceptible animals. The resistant group presented higher mean packed cell volume (26.48%) and total plasma protein (6.24 g / dl) than susceptible one (24,04% e 5,82g/dl, respectively). The average number of eosinophils was similar in both groups The Haemonchus sp. was the most prevalent in the culture of feces, followed by Trischostrongylus sp. and Oesophagostomum sp.. The counting of nematodes in the abomasum of susceptible group was higher than in resistant one. The species identified were H. contortus in abomasums and T. colubriformis in small intestine. It can be concluded that EPG, packed cell volume and total plasma protein were useful phenotypic markers to identify animals as resistant and susceptible to gastrointestinal nematodes infections

Fatores de influência na produção de leite bovino sobre os níveis da contagem bacteriana total

The objective of this study was to evaluate the influence of milking procedures on the levels of total bacterial count (TBC) in bovine milk. In the first study the influences of procedures for hygienic milking, cleaning of milking equipment and milk cooling tanks on the TBC levels were evaluated. Four bulk samples of milk were collected from each tank in eight properties for TBC analysis, employing the flow cytometry method. A questionnaire was applied in each property to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology for analysis of longitudinal data was considered, focusing on random effects models. The results showed that the handling procedures for milking and the cleanliness of the cooling tank contributed to a further reduction in the levels of TBC raw milk cooling tanks. The second study aimed to describe the percentage of the properties that comply with the Normative Instruction Nº 51 (Brazil s IN 51) with regard to total bacterial count (TBC) in bovine milk. The study was conducted from January 2010 to July 2011. Milk samples were collected from the eight properties selected for TBC analysis by the flow cytometry method. Again, on each property a questionnaire was applied to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology of marginal models based on Generalized Estimate Equations (GEEs) was followed in the statistical analysis. The results showed that the handling procedures of the milking and the cleanliness of the cooling tanks contributed to a considerable percentage of the properties that reached the limits of TBC established by IN 51

Estimação de parâmetros genéticos para resistência à infecção por IMNV em camarões Litopenaeus vannamei por meio de análise de sobrevivência

The main specie of marine shrimp raised at Brazil and in the world is Litopenaeus vannamei, which had arrived in Brazil in the `80s. However, the entry of infectious myonecrosis virus (IMNV), causing the infectious myonecrosis disease in marine shrimps, brought economic losses to the national shrimp farming, with up to 70% of mortality in the shrimp production. In this way, the objective was to evaluate the survival of shrimps Litopenaeus vannamei infected with IMNV using the non parametric estimator of Kaplan-Meier and a model of frailty for grouped data. It were conducted three tests of viral challenges lasting 20 days each, at different periods of the year, keeping the parameters of pH, temperature, oxygen and ammonia monitored daily. It was evaluated 60 full-sib families of L. vannamei infected by IMNV in each viral challenge. The confirmation of the infection by IMNV was performed using the technique of PCR in real time through Sybr Green dye. Using the Kaplan-Meier estimator it was possible to detect significant differences (p <0.0001) between the survival curves of families and tanks and also in the joint analysis between viral challenges. It were estimated in each challenge, genetic parameters such as genetic value of family, it`s respective rate risk (frailty), and heritability in the logarithmic scale through the frailty model for grouped data. The heritability estimates were respectively 0.59; 0.36; and 0.59 in the viral challenges 1; 2; and 3, and it was also possible to identify families that have lower and higher rates of risk for the disease. These results can be used for selecting families more resistant to the IMNV infection and to include characteristic of disease resistance in L. vannamei into the genetic improvement programs