Processo biotecnológico voltado a produção de estoques poliploides do camarão-branco litopenaeus vannamei (decapoda, penaeidae)

without practical results so far. Protocols used in biotechnological cultured aquatic organisms aimed at increasing growth rates and disease resistance, have been studied and perfected. Among the available techniques, the application of chromosomal manipulation, although still nascent, is presented as a tool aimed at mitigating ecological and economical issues in shrimp farming. The polyploidization artificial method already employed in fish and shellfish, has been widely researched for use in farmed shrimp. Some limitations of this method of expansion in shrimp refer to a better knowledge of cytogenetic aspects, the level of sexual dimorphism and performance in growing conditions. To contribute on some of these issues, the present study aimed to characterize cytogenetic species Litopenaeus vannamei (Decapoda) and Artemia franciscana (Anostraca), analyze the effectiveness of methods for detection of ploidy, through the use of flow cytometry in processes of induction polyploidy cold thermal shock at different stages of development of newly fertilized eggs. Additionally, aimed also the qualitative and quantitative comparison of larval development between diploid and polyploid organisms, besides the identification of sexual dimorphism in L. vannamei, through geometric morphometrics. The results provide information relevant to the improvement and widespread use of biotechnological methods applied toward national productivity in shrimp farming

Processo biotecnológico voltado a produção de estoques poliploides do camarão-branco litopenaeus vannamei (decapoda, penaeidae)

without practical results so far. Protocols used in biotechnological cultured aquatic organisms aimed at increasing growth rates and disease resistance, have been studied and perfected. Among the available techniques, the application of chromosomal manipulation, although still nascent, is presented as a tool aimed at mitigating ecological and economical issues in shrimp farming. The polyploidization artificial method already employed in fish and shellfish, has been widely researched for use in farmed shrimp. Some limitations of this method of expansion in shrimp refer to a better knowledge of cytogenetic aspects, the level of sexual dimorphism and performance in growing conditions. To contribute on some of these issues, the present study aimed to characterize cytogenetic species Litopenaeus vannamei (Decapoda) and Artemia franciscana (Anostraca), analyze the effectiveness of methods for detection of ploidy, through the use of flow cytometry in processes of induction polyploidy cold thermal shock at different stages of development of newly fertilized eggs. Additionally, aimed also the qualitative and quantitative comparison of larval development between diploid and polyploid organisms, besides the identification of sexual dimorphism in L. vannamei, through geometric morphometrics. The results provide information relevant to the improvement and widespread use of biotechnological methods applied toward national productivity in shrimp farming

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

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

Moléculas coestimulatórias na leishmaniose visceral

Visceral leishmaniasis (VL) is endemic in many countries, including Brazil. The protozoan Leishmania infantum, is the etiological agent of VL, and is transmitted by the bite of female sandflies during the blood meal. The majority of subjects when exposed to the parasite do not develop the disease, because of development of Th1 cellular responses. Those who have develop signs of VL such as fever, weight loss, hepatosplenomegaly, have impairment of the cellular immune response, specific to the Leishmania antigens. We evaluated whether the specififc anergy during symptomatic VL, may be associated with changes in T cells costimulatory molecules or their ligands in CD14+ monocytes. There is an increase in CTLA-4 porcentage on CD4+ T lymphocytes (p=0.001) and ICOS on CD4+ and CD8+ T cells (p=0.002 to CD4+ and p=0.003 to CD8+), after stimulation by soluble Leishmania antigen (SLA) during active visceral leishmaniasis, and that there is a higher percentage of these molecules ex vivo, when comparing symptomatic to recovered individuals (p=0.04 to CTLA-4 in CD4+, and p=0.001 to ICOS in CD4+ and p=0.026 to CD8+). Moreover, we found a high gene expression of CTLA-4, OX-40 and ICOS during active VL. CD40, CD80, CD86, HLA-DR and ICOSL molecules do not suffer changes during disease. There is IFN-γ production by the peripheral blood cells, after SLA stimulation, by peripheral blood cells in symptomatic subjects; however, there is a decrease of the ratio IFN-γ/IL-10, which is reversed after clinical recovery. The impairment of some costimulatory molecules pathways during symptomatic VL could inhibit the ability of phagocytes to kill Leishmania and could facilitate their survival and the proliferation inside macrophages.

Avaliação do desenvolvimento infantil e a influência dos fatores biopsicossociais em crianças com cardiopatia congênita

Child development is the result of the interaction of biological, psychological and social factors. Hostile environment, income, offered stimuli, as well as the presence of a chronic illness are issues that may interfere significantly. Considering the chronic diseases, we can identify congenital heart disease (CHD) is characterized by anatomical heart defects and functional and currently has presented an incidence of up to 1% of the population of live births. This research aimed to evaluate child development and verify an association with the commitment by biopsychosocial factors of children with and without CHD. Study participants were children from zero to six years, divided into three groups: Group1- 29 children pre-surgical congenital heart disease, Group2- 43 children post-surgical cardiac patients and Group3- 56 healthy children. The instruments used were a biopsychosocial questionnaire and the Screening Test Denver II. Of the total of 128 children evaluated, 66 (51.56%) are girls, and ages ranged from two months to six years (median 24.5 months). In G1 and G2 predominated acyanotic heart disease (55.2% and 58.1%). Regarding the Denver II reviews, children with heart disease had more development ratings "suspicious" and "suspect/abnormal", and 41.9% of children who have gone through surgery had characterized its development as "suspect/abnormal" . In the group of healthy children 53.6% were classified as developmental profile "normal" (p = ˂0,0001). On the areas of Denver II, among children with heart disease was greatest change in motor areas (p = 0.016, p = ˂0,001). The biopsychosocial variables that were related to a possible developmental delay were gender (p = 0.042), child's age (p = 0.0001) and income per capita (p = 0.019). There were no associations between the variables related to the treatment of disease, information, understanding of the disease and the way parents treat their children. In the group of healthy children showed that children who underwent hospitalization rates were more changes in development (p = 0.025) and the higher the number of admissions over these changes have intensified (p = 0.023). The results suggest that children with congenital heart disease have likely delayed development. It was also observed that there is a significant difference between the children who have gone through surgery, those who are still waiting for surgery only doing clinical follow-up. Changes in the development are more connected motor areas can be explained by the characteristic features of the disease and treatment, such as dyspnea, fatigue, care and limitations in daily activities. The gender and age appear to be decisive in the development as well as healthy children go through hospitalization experience. Already in children with heart disease, it was realized that social variables involved in the disease and the treatment did not affect the development. This question can be understood by means of protective factors and resiliency, as this population receives family and social support.