Influência do vírus da hepatite G (GBV-C) na resposta imune frente à infecção por Leishmania chagasi

GB virus type C (GBV-C) appears to promote a Th1 response and is associated with prolonged survival in HIV-infected people. L. chagasi causes a spectrum of illness that varies from severe visceral leishmaniasis, a disease that in the majority of cases is fatal if not treated, to self resolution of infection and development of positive DTH response that is protective against symptomatic disease. To determine if GBV-C viremia might influence the outcome of Leishmania infection, we characterized GBV-C status in a cohort of subjects residing in a L. chagasi endemic area in Brazil. GBV-C viremia was more prevalent in blood donors from urban than in periurban regions of Natal, Brazil (16% and 7.5% respectively). Evidence of prior GBV-C (anti-E2 antibodies) was detected in 24% and 12%of these groups respectively. Anti-E2 increased with age (p= 0.0121). No difference in GBV-C viremia was found in the DTH+ and VL groups (p= 0.269); however, subjects with visceral leishmaniasis were more likely to have anti-E2 than DTH+ subjects (p=0.0012), and DTH induration was smaller in subjects with E2 antibodies (4.5 mm) compared those without (7.12 mm) (p= 0.002). Furthermore, the size of the Leishmania DTH response was greater in GBV-C viremica subjects (6.8 mm) compared to non-viremic subjects (3.3 mm; p= 0.0054). There findings suggest that GBV-C virus may promote a type 1 immune response that could influence the outcome of Leishmania infection

Co-infecção pelo vírus da hepatite G em pacientes infectados pelo Leihsmania chagasi

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Estudo do genoma do vírus causador da mionecrose infecciosa em camarões e desenvolvimento de métodos para detecção de diferentes variantes

Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites

Influência de fatores ambientais na incidência do vírus da infecção hipodermal e necrose hematopoiética (IHHNV) no camarão Litopenaeus vannamei cultivado em fazendas do Estado do Rio Grande do Norte (RN)

The shrimp farming industry is the most profitable area of the aquaculture at Rio Grande do Norte (RN) state, which is one of the largest producers in Brazil. However the infections that affect the shrimp cause major economic losses. The infection is a result of the interaction between the shrimp, the environment and pathogen. The change of these factors may lead to a condition of stress and susceptibility to opportunistic infections. One of these infections caused by Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) is widely distributed in several countries and affects a wide range of hosts. To optimize conditions for production of Litopenaeus vannamei shrimp, the more species cultivated in Brazil, it is necessary to understand the effects of environmental factors in the susceptibility of this species to infections. The aim of this study was to determine the IHHNV prevalence and to investigate the influence of environmental factors as salinity, temperature, stocking density, dissolved oxygen and rainfall in the IHHNV incidence in L. vannamei grown in farms, in the RN state. To determine the IHHNV prevalence were used 1089 samples of L. vannamei collected in seven farms. To perform the study about the influence of environmental factors, 525 samples of L. vannamei shrimp were collected in eight farms located in regions of low (0-1 ), medium (21-30 ) and high (38-57 ) salinity, using extensive (≤15 shrimp/m2 ), semi-intensive (18-33 shrimp/m2) or intensive (>36 shrimp/m2) stocking density systems. The IHHNV infection was determined in pleopod and hemolymph using the polymerase chain reaction (PCR). The environmental factors were recorded during the collection of animals, using a refractometer to measure the salinity and a multi-parameter meter to measure the temperature and concentration of dissolved oxygen in the water. The IHHNV prevalence in RN was 43% (468 infected shrimp out of 1089), varying on different farms. On the seven farms studied, IHHNV prevalence ranged from 18.6% to 54.8%. The infection rates in the shrimp cultured in low, medium and high salinity were respectively 43.10% (125/290), 31.2% (15/48) and 24.6% (46/187) and was significantly higher in shrimp grown in low salinity (P<0.001). The infection rates in ponds of extensive, semi-intensive and intensive systems were respectively, 28.7%, 28.28% and 47.84%, and was significantly higher in high stocking densities (P<0.001). This study indicated a high IHHNV prevalence and a significant effect of salinity and stocking density, but not of the temperature, rainfall and dissolved oxygen on the IHHNV infection rate in the L. vannamei shrimp cultured in the northeastern Brazil

Prevalência da infecção genital pelo vírus herpes simples tipo 1 e 2 em mulheres grávidas e não grávidas de Natal/RN

Herpes simplex is a virus that can be transmitted sexually and is potentially associated with vertical transmission. This study evaluated the prevalence of genital infection by herpes simplex virus (HSV) types 1 and 2 in pregnant and nonpregnant care in the city of Natal / RN, including a total of 222 women, 92 pregnant and 130 nonpregnant. The participants answered a questionnaire to obtain data and socio-demographic characteristics, as well as potential risk factors for sexually transmitted diseases. After the interview, we collected two cervical specimens, one for the Pap test and the other for DNA extraction and analyzed by polymerase chain reaction (PCR) to detect both virus serotypes. Then the women underwent a clinical examination by colposcopy. For statistical analysis, we used the chi-square and logistic regression by SSPS 17.0 Statistic. Most women were up to 30 years of age, nonwhite ethnicity, married, elementary education, family income below the poverty level; initiated sexual activity with age up to 18 years; had more than one sexual partner lifelong and was not pregnant, but has had at least one child. The HSV-1 showed a prevalence of 26.1% among pregnant women and 30.0% in non-pregnant women. While HSV-2 prevalence was found with 10.9% and 19.2% in pregnant and nonpregnant women, respectively. The largest proportion of morphological changes of the uterine cervix was detected among nonpregnant women, both in cytology and in colposcopy. The women were nonwhite ethnicity, married, became pregnant aged less than or equal to 18 years and who had one to two pregnancies had a lower risk of acquiring genital HSV infection. There was a high prevalence of genital HSV infection, HSV-1 is more prevalent than HSV-2. No association was found between morphological changes of the uterine cervix and the presence of the virus in pregnant and nonpregnant women, nor between genital HSV infection and the classic risk factors for sexually transmitted diseases

Correlação entre a infecção genital pelo vírus do papiloma humano, a resposta imune e os achados colpocitológicos em mulheres grávidas e não grávidas

The genital HPV infection is very common between men and women worldwide, affecting particularly young women, constituting a serious public health problem in less developed regions, favored by the poor living conditions of population. The cytology and colposcopy have notorious importance in the diagnosis of precursor lesions of cervical cancer and therefore its prevention. However, even with such diagnostic tools, the number of women who develop cervical cancer is still high. This study aims to assess the prevalence of genital tract infection by HPV in pregnant and nonpregnant women, evaluating the profile of the immune response presented by the women of these two groups in order to establish correlations among profile of immune response, presence of virus and occurrence of lesions of the uterine cervix. We analyzed specimens obtained from the cervix of 221 patients, 91 pregnant and 130 non-pregnant, aged 14-72 years. The women were subjected to colposcopic and cytologic evaluation detect possible changes in the cervix and then samples were collected in order to perform HPV detection by PCR and real-time PCR for detection of mRNA of pro-inflammatory and anti-inflammatory cytokines. In the present study, the overall prevalence of HPV genital infection was 28.1%; of which 31.9% were pregnant patients and 25.4% in non-pregnant women. Young women under 30 years and those with low educational level education showed a higher risk of HPV infection. Colposcopy showed better correlation with detection of HPV DNA by PCR, when compared to cytology. Generally, HPV infected patients, pregnant or not, exhibited reduced mRNA expression of both pro-inflammatory (IFN-γ, TNF-α) and anti-inflammatory (IL -10) cytokines, when compared to patients not infected by HPV. Nonpregnant patients infected presented increase mRNA expression of IL-17 in patients without injury, whereas those with lesion showed higher mRNA expression of TGF-β. Pregnant women without injury infected exhibited increased mRNA expression of TGF-β. There was no difference in HPV prevalence between pregnant and nonpregnant women. There was a reduction of pro-inflammatory cytokines, except IL-17, in all women infected by HPV. Moreover, we observed an increase of TGF-β in HPV-infected women who are pregnant or not. The results suggest that, in women in this study, HPV infection promoted changes in the profile of cytokines necessary for activation of effective immune response, possibly favoring viral persistence

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

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