Avaliação preliminar da viabilidade de produção in vitro de um isolado brasileiro de baculovírus Spodoptera frugiperda MNPV

In vivo production of viral biopesticides is the major source of viral insecticides currently in the marketplace. However, this system presents limitations during production scale-up. For the Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV), the insect used for replication has cannibalistic characteristics, thus production is even more difficult. Insect cells are commonly used for in vitro baculovirus production. Most of these cell lines are derived from Lepidoptera species. The Sf21 cell line is derived from Spodoptera frugiperda caterpillar ovarian tissue, and its clonal isolate Sf9 has been used for biopesticide production due to its ease of growth in suspension cultures. In this work, the in vitro production capabilities of a Brazilian SfMNPV isolate obtained from cornfields was evaluated. Comparison of polyhedra production was carried out using both Sf21 and Sf9 cells, based on volumetric and specific yields. Both cell lines were cultivated in Hyclone medium supplemented with different fetal bovine serum concentrations (2,5 and 5%). The best results were obtained using Sf9 cells supplemented with 5% serum. These results were further confirmed quantitively through kinetic parameter estimation for both cells lines and different serum concentrations. After seven successive passages, this system still presented high specific polyhedra production

Bidens mosaic virus: detecção via RT-PCR e identificação de Galinsoga parviflora como um novo hospedeiro natural do vírus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resposta de plantas de alho livres de vírus ao nitrogênio em ambiente protegido

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos efeitos agudos de diferentes niveis de PEP sobre o padrão respiratório e volumes operacionais de pacientes com Parkinson

Objetivo: Determinar os efeitos agudos de diferentes intensidades de pressão expiratória positiva sobre o padrão respiratório e volumes operacionais de pacientes com doença de Parkinson. Métodos: Foram eleitos para o estudo 23 pacientes em estágios II ou III da doença, estando na condição ON , sendo 8 excluídos, e 15 controles saudáveis. Os indivíduos foram submetidos a uma avaliação inicial, constando de dados gerais, avaliação antropométrica, função pulmonar e força muscular respiratória. Após esta etapa inicial, realizou-se a avaliação dos volumes pulmonares com pletismografia optoeletrônica associado a utilização de pressão expiratória positiva (PEP) em três intensidades de pressão positiva, 10 cmH2O, 15 cmH2O e 20 cmH2O em ordem aleatória. Resultados: O grupo Parkinson apresentou valores espirométricos e de força muscular respiratória significativamente menores que o grupo controle (p<0.01). Houve diferença nos valores de volume corrente (Vt) do grupo Parkinson em relação ao grupo controle na respiração tranqüila (p<0.001) e aumento no Vt do grupo Parkinson com uso das três intensidades de PEP (p<0.001), sem diferenças estatisticamente significativas entre as intensidades e com diferença na distribuição compartimental do Vt entre os grupos (p<0.001). O fluxo inspiratório médio e o volume minuto do grupo Parkinson foram menores em relação ao grupo controle na respiração tranqüila (p<0.001) e aumentaram com o uso das três intensidades de PEP (p<0.001), sem diferenças estatisticamente significativas entre as intensidades. Houve diferença na variação dos volumes operacionais entre os grupos em todas as intensidades de PEP (p < 0.001). O volume pulmonar expiratório final não aumentou no grupo Parkinson com uso de PEP. O volume pulmonar inspiratório final aumentou no grupo Parkinson em todas intensidades de PEP (p < 0.001), não havendo diferenças entre as intensidades. Conclusão: Os pacientes com doença de Parkinson apresentam alterações em diferentes componentes do padrão respiratório e a terapia com PEP determina modificações clinicamente importantes nos volumes pulmonares, considerando a intensidade de 10 cmH2O suficientes para este objetivo terapêutico

Estudo da detecção do DNA do papiloma vírus humano (HPV) e da expressão imuno-histoquímica de proteína do ciclo celular no carcinoma epidermóide oral

Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease

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

Caracterização dos sistemas de produção de leite bovino na microrregião Seridó do estado do Rio Grande do Norte

The objective was to contribute to a reading of the characteristics and diversity of production systems from bovine milk in the microregion Seridó of Rio Grande do Norte, discuss the social, economic, environmental and husbandry-related primary activity of milk production. We randomly selected 28 agricultural establishments that performed the activity of dairy bovine culture with subsequent applicat ion of a structured questionnaire during September and October 2011. Data were analyzed with application of measures of descriptive analysis and determination of the index rural development (IRD). The results showed that 53.57% of the interviewees were owners of the land, the median area of the properties amounted to 135 hectares, the median number of animals in the herd was 51 head, with minimum 11 and maximum of 350 heads establishments in the sample, 85.72% of establishments had maximum 23 cows in lactation, 100% performed manual milking with suckling calves, average productivity of 3.91 liters / cow / day, 92.86% of the interviewees did not produce silage and / or hay, 64.29% had no access to technical assistance, the average age of interviewees was 51 ± 10.85 years, 78.57% had only elementary education. The average of the IRD computed in the test sample amounted to 0.43 ± 0.11 on a scale 0-1. The sites were grouped into 04 classes, based on your IRD. The average IRD in each dimension was 0.43, 0.55 and 0.34, respectively for economic / husbandry, social and environmental. The determination of the rate of rural development - IDR found for the sample could add establishments hierarchically. It is necessary that the inclusion of best practices in herd management, bookkeeping zootechnical, technical assistance and reorganization practices in land and environmental preservation

Levantamento exploratório da composição e frações proteicas do leite bovino em propriedades leiteras

The direct relationship with the processing, industrial yield and price of milk, the importance of the centesimal composition of milk and their protein fractions has increased for both the dairy industry and for the producers, so it is important to identify the possible causes on variation of these components. The aim of this study was to evaluate the effect of the seasons, order of calving and lactation phase on milk composition and protein fractions in dairy farms in the Agreste and Leste meso-regions, of Rio Grande do Norte State (RN). Milk samples were taken in seven milk farms directly from the bulk tanks except for one farm where samples were taken from higher milking cows producers and heifers. The experiment was done in seven properties with milk samples taken directly from the bulk tanks and in one property were collected from higher production cows and heifers. Composition analyses were carried out for both experiments and sensorial trial was performed for the second experiment only. The data results were submitted to analysis of variance (ANOVA) and Tukey test performed for 5% significance level. In the first experiment was observed response of the seasons, dry and rainy season; while in the second, the effect of the order of calving and lactation phase had influence on milk composition and protein fractions. There was significant response (p < 0.05) for season effects but due to farms. The rainy season had higher averages in the protein, lactose, total solids and nonfat dry extract. Multiparous cows had higher CCS, NUL and PCAS milk contents; while, first calving heifers showed higher average for the sensory attribute, color of milk. The final third of lactation was responsible for the elevation of the levels of almost all milk components and their protein fractions

Avaliação da estabilidade térmica e oxidativa dos biodieseis de algodão, girassol, dendê e sebo bovino

The search for new sources of environmentally friendly energy is growing every day. Among these alternative energies, biodiesel is a biofuel that has had prominence in world production. In Brazil, law 11.097, determine that all diesel sold in the country must be made by mixing diesel/biodiesel. The latter called BX, , where X represents the percent volume of biodiesel in the diesel oil, as specified by the ANP. In order to guarantee the quality of biodiesel and its mixtures, the main properties which should be controlled are the thermal and oxidative stability. These properties depend mainly of the chemical composition on the raw materials used to prepare the biodiesel. This dissertation aims to study the overall thermal and oxidative stability of biodiesel derived from cotton seed oil, sunflower oil, palm oil and beef tallow, as well as analyze the properties of the blends made from mineral oil and biodiesel in proportion B10. The main physical-chemical properties of oils and animal fat, their respective B100 and blends were determined. The samples were characterized by infrared and gas chromatography (GC). The study of thermal and oxidative stability were performed by thermogravimetry (TG), pressure differential scanning calorimeter (PDSC) and Rancimat. The obtained biodiesel samples are within the specifications established by ANP Resolution number 7/2008. In addition, all the blends and mineral diesel analyzed presented in conformed withthe ANP Regularion specifications number 15/2006. The obtained results from TG curves data indicated that the cotton biodiesel is the more stable combustible. In the kinetic study, we obtained the following order of apparent activation energy for the samples: biodiesel from palm oil > sunflower biodiesel > tallow biodiesel > cotton biodiesel. In terms of the oxidative stability, the two methods studied showed that biodiesel from palm oil is more stable then the tallow. Within the B100 samples studied only the latter were tound to be within the standard required by ANP resolution N° 7. Testing was carried out according to the EN14112. This higher stability its chemical composition

Caracterização de resíduo sólido formado em biodiesel de sebo bovino

Biodiesel production has increased over the last decade because of the benefits associated with this fuel, including renewability, domestic feedstock, lower toxicity, and biodegradability. From 2008, the use of beef tallow as a feedstock for biodiesel production in Brazil has increased in significance, representing the second largest source of biodiesel, after soybeans. However, the performance of biodiesel in cold weather conditions is worse than diesel because of deposition of insoluble at low temperatures, accelerating the plugging of fuel filters and injectors of the vehicle engine. Studies have been conducted on beef tallow biodiesel, mostly related to the properties of thermal and oxidative stability. However, few studies have described the nature of the precipitate formed and its influence on product quality. Research suggests that the cause of deposition is related to the nature of saturated esters and monoacylglycerols as inducing agents. This study monitored the levels of mono-, diand triacylglycerols, the oxidation stability and the cold filter plugging point (CFPP) in beef tallow biodiesel samples from two commercial producers in Brazil for a period of twelve months. Filtered precipitates were analyzed by comparative techniques of GCFID, HPLC-UV/VIS, HPLC-MS-IT-TOF and TG to verify the nature, using monopalmitin and monostearin as reference standards. The formation of precipitate reduced the levels of monoacylglycerols in the beef tallow biodiesel. GC-FID and LCMS- IT-TOF results confirmed the nature of the deposit as saturated monoacylglycerols, predominantly monostearin and monopalmitin as the second major component. Moreover the TG analysis of the residue indicated similar thermal decomposition of the reference standards. The precipitate did not affect the oxidation stability of beef tallow biodiesel and the CFPP characteristic of blends up B60. However, the presence of iron reduced significantly the oxidation stability of biodiesel

Caracterização de resíduo sólido formado em biodiesel de sebo bovino

Biodiesel production has increased over the last decade because of the benefits associated with this fuel, including renewability, domestic feedstock, lower toxicity, and biodegradability. From 2008, the use of beef tallow as a feedstock for biodiesel production in Brazil has increased in significance, representing the second largest source of biodiesel, after soybeans. However, the performance of biodiesel in cold weather conditions is worse than diesel because of deposition of insoluble at low temperatures, accelerating the plugging of fuel filters and injectors of the vehicle engine. Studies have been conducted on beef tallow biodiesel, mostly related to the properties of thermal and oxidative stability. However, few studies have described the nature of the precipitate formed and its influence on product quality. Research suggests that the cause of deposition is related to the nature of saturated esters and monoacylglycerols as inducing agents. This study monitored the levels of mono-, diand triacylglycerols, the oxidation stability and the cold filter plugging point (CFPP) in beef tallow biodiesel samples from two commercial producers in Brazil for a period of twelve months. Filtered precipitates were analyzed by comparative techniques of GCFID, HPLC-UV/VIS, HPLC-MS-IT-TOF and TG to verify the nature, using monopalmitin and monostearin as reference standards. The formation of precipitate reduced the levels of monoacylglycerols in the beef tallow biodiesel. GC-FID and LCMS- IT-TOF results confirmed the nature of the deposit as saturated monoacylglycerols, predominantly monostearin and monopalmitin as the second major component. Moreover the TG analysis of the residue indicated similar thermal decomposition of the reference standards. The precipitate did not affect the oxidation stability of beef tallow biodiesel and the CFPP characteristic of blends up B60. However, the presence of iron reduced significantly the oxidation stability of biodiesel

Incidência e severidade do vírus do endurecimento dos frutos em maracujazeiros enxertados em pé-franco

O objetivo deste trabalho foi avaliar a incidência e a severidade do vírus do endurecimento dos frutos em maracujazeiro-amarelo enxertado e pé-franco. O experimento foi conduzido no município de Adamantina-SP, no período de abril de 2006 a junho de 2007, adotando-se o delineamento em blocos ao acaso, com quatro tratamentos e oito repetições. Foram avaliados três porta-enxertos: Passiflora edulis, P. alata e P. gibertii, e plantas de pé-franco. Utilizou-se como copa o maracujazeiro-amarelo (Passiflora edulis Sims). Avaliaram-se a porcentagem de plantas com sintomas de virose e a severidade dos sintomas. As primeiras plantas com sintomas de virose ocorreram aos 90 dias do plantio das mudas no campo, atingindo, aos 180 dias, 100% de plantas com virose em P. alata e P. gibertii, e 97,5% em P. edulis e pé-franco.

Comparação entre pericárdio bovino preservado em glicerina e malha de poliéster no reparo de falhas da parede abdominal em ratos

OBJETIVO: Comparar uma malha comercial de poliéster com o pericárdio bovino preservado em glicerina na reconstituição de defeitos da parede abdominal. MÉTODOS: Foram utilizados 30 ratos, divididos em dois grupos eqüitativos. Efetuou-se uma excisão retangular de 2,5 x 2 cm, incluindo toda a musculatura abdominal e peritônio. No grupo I a parede abdominal foi reparada com malha de poliéster e no grupo II com pericárdio bovino conservado em glicerina. Os animais foram sacrificados aos 15, 60 e 90 dias de pós-operatório, sendo o local cirúrgico avaliado macroscópica e histologicamente. RESULTADOS: Os animais do grupo I apresentaram aderências mais severas e em maior número quando comparados aos do grupo II; porém, sem comprometimento funcional. A análise histológica revelou incorporação dos tecidos aos implantes, com maior resposta fibroblástica nos animais do grupo I. CONCLUSÃO: A malha de poliéster oferece maior resistência estrutural e resposta fibroblástica mais intensa; contudo, promove grande quantidade de aderências às vísceras abdominais, quando comparada ao pericárdio bovino.

Caracterização genética e evolução dos vírus dengue no Estado do Rio Grande do Norte

Dengue is considered as the most important arthropod-borne viral disease throughout the world due to the high number of people at risk to be infected, mainly in tropical and subtropical regions of the planet. The etiologic agent is Dengue Virus (DENV), it is a single positive-stranded RNA virus of the family Flavivirus, genus Flaviviridae. Four serotypes are known, DENV-1, DENV-2, DENV-3 and DENV-4. One of the most important characteristic of these viruses is the genetic variability, which demands phylogenetic and evolutionary studies to understand key aspects like: epidemiology, virulence, migration patterns and antigenic characteristics. The objective of this study is the genetic characterization of dengue viruses circulating in the state of Rio Grande does Norte from January 2010 to December 2012. The complete E gene (1485 pb) of DENV1, 2 e 4 from Brazilian (Rio Grande do Norte) patients was sequenced. Phylogenetic analysis was performed using MEGA 5.2 software, Tamura-Nei model and Neighbor-Joining trees were inferred for the datasets. In Brazil, there is just one DENV-1 genotype (genotype V), one DENV-2 genotype (Asian/American) and two DENV-4 genotypes (genotypes I and II). Brazilian strains of DENV-1 are subdivided in two different lineages (BR-I and BR-II), the Brazilian strains of DENV-2 are subdivided in four lineages (BRI-IV) and genotype II of DENV-4 is subdivided in three Brazilian lineages (BRI-III). The viruses isolated in RN belong to lineage BR-II (DENV-1), BR-IV (DENV-2) and BR-III (DENV-4).The Caribbean and near Latin American countries are the main source of these viruses to Brazil. Amino acids substitutions were detected in three domains of E protein, this makes clear the necessity of studies that associate epidemiological and molecular data to better understand the effects of these mutations. This is the first study about genetic characterization and evolution of Dengue viruses in Rio Grande do Norte, Brazil

Hibridização reversa e sequenciamento na genotipagem do vírus da hepatite C

INTRODUÇÃO: Os métodos de genotipagem do vírus da hepatite C têm sido muito discutidos. O objetivo deste trabalho foi comparar as metodologias de hibridização reversa e sequenciamento direto para a genotipagem do vírus da hepatite C. MÉTODOS: Noventa e uma amostras de plasma de pacientes assistidos na Faculdade de Medicina de Botucatu da Universidade Estadual Paulista foram utilizadas. A genotipagem por hibridização reversa foi realizada utilizando o kit comercial INNO-LiPA® v.1.0. O sequenciamento direto foi efetuado em sequenciador automático utilizando protocolos in house. RESULTADOS: A genotipagem por sequenciamento direto mostrou-se eficiente na resolução dos resultados inconclusivos pelo kit comercial. O kit mostrou resultados errôneos em relação à subtipagem viral. Além disso, a genotipagem por sequenciamento direto revelou um erro do kit com relação à determinação genotípica questionando a eficiência do método também para a identificação do genótipo viral. CONCLUSÕES: A genotipagem realizada por meio de sequenciamento direto permite uma maior acurácia na classificação viral quando comparada à hibridização reversa.