Diversity of cellulolytic and xylanolytic fungi associated with the digestive tract of aquatic insect larvae in streams of the Amazon Forest and Cerrado in Brazil.

The study of the relationship between fungi and insects brings important contributions to the knowledge of fungal biodiversity and to the understanding of mutualistic ecological interactions. This study reports the occurrence of a community of filamentous fungi in the digestive tract (DT) of mining insect larvae belonging to genus Stenochironomus in streams of two Brazilian biomes. Fungi were obtained from the digestive tract of larvae found on trunks and leaves of low-order streams in the Amazon Forest and Cerrado in the north of Brazil. The fungal community was screened for xylanolytic and cellulolytic activities. The diversity of fungal species in the DT of larvae is possibly related to the diversity of diets of species of that genus and the diversity of substrates in the ecosystems. The diversity and richness of fungal species were influenced by ecological differences between locations more than by the types of substrates in which they were collected (trunk and leaf). Most fungi in the DT of Stenochironomus larvae sampled in leaves exhibited cellulolytic enzyme activity. Such results stress that the mycobiomes of the DT of Stenochiromonus larvae produce enzymes that contribute to the process of breaking down plant remains in their hosts.

Diversity and enzymatic capabilities of fungi associated with the digestive tract of larval stages of a shredder insect in Cerrado and Amazon Forest, Brazil.

Tropical biomes such as Brazilian Cerrado and Amazon Forest have a great diversity of fungi and insects. Interactions between these organisms can be beneficial to both partners. In streams, these interactions contribute to litter decomposition. Studying the digestive tract (DT) of shredder insects as a habitat for fungal microorganisms is an opportunity to obtain fungal strains with biotechnological potential, which may help to understand the symbiotic relationships between these organisms in tropical forests. This study investigated the fungal community in the DT of larvae of Triplectides (Trichoptera: Leptoceridae) collected in low-order streams in the Cerrado and Amazon Forest biomes in Brazil. Forty-nine fungal isolates were obtained and identified among 32 species and 12 genera. The genus Roussoella was only found in the DT of insects in Amazon Forest streams, while 7 genera only occurred in the DT of insects in Cerrado streams. The genus Penicillium (40%) was the most frequent. In the Cerrado, 78% were producers of CMCase, more than two-fold that in the Amazon Forest (35%). And 62% were producers of xylanase, in the Cerrado and 71% in the Amazon Forest. In this context, the fungal community in the DT of Triplectides larvae may play an important role in the insect diet by breaking down lignocellulosic material.

Production of low-cholesterol butter with Lacticaseibacillus paracasei immobilized in calcium-alginate beads.

This study focused on the application of three strains of Lacticaseibacillus paracasei to assimilate cholesterol in cream and butter. The strains were enclosed in calcium-alginate beads and incubated in cream at 30 °C for 15 h. Immobilization of lactobacilli cultures in calcium-alginate beads resulted in a 23% reduction in cholesterol (p < 0.05) in cream, whereas a negligible reduction was observed in cream fermented with free cells. Butter with a 44% reduction in cholesterol was produced from fermented cream by L. paracasei L2A21K5 entrapped in alginate beads. No significant (p > 0.05) changes in the fatty acid profile were observed in the low-cholesterol butter, except for a slight but significant increase in n-3 fatty acids (p < 0.05). In addition, the indices of atherogenicity and thrombogenicity were significantly reduced in the low-cholesterol butter (p < 0.05). Panelists rated the low-cholesterol butter as good in appearance, consistency, and flavor.

Avaliação de um controle estratégico da artrite encefalite caprina em rebanho caprino leiteiro / Evaluation of strategic control of caprine arthritis- encephalitis in dairy herd goats

O objetivo deste trabalho foi avaliar a utilização periódica de testes de diagnóstico mais sensíveis aliados às práticas de manejo, visando ao controle eficaz da artrite encefalite caprina (CAE). Foram realizadas oito coletas de sangue em matrizes e reprodutores. Da primeira à sétima análise, as coletas foram quadrimestrais, utilizando-se os testes de imunodifusão em gel de agarose (IDGA), ensaio imunoenzimático indireto (ELISA-i) e Western Blot (WB). A oitava coleta aconteceu seis meses após a sétima, utilizando-se o WB e a reação em cadeia de polimerase (PCR). A prevalência da CAE foi de 6,8%, 14,9% e 39,2% no IDGA, ELISA-i e WB, respectivamente. Na última análise, foram detectados 0,9% de animais positivos pelo WB e 10,8% pela PCR. Apesar de não erradicarem a CAE, as medidas adotadas, aliadas à utilização periódica dos testes sorológicos e à combinação com a PCR, foram importantes para reduzir significativamente os animais soropositivos no rebanho.(AU)

The aim of this study was to evaluate the periodic use of more sensitive diagnostic tests associated to management practices for the effective control of caprine arthritis-encephalitis (CAE). We carried out eight blood samples in does and bucks. From the first to the seventh analysis, the samples were quarterly, using Agarose Gel Immunodiffusion (AGID), Enzyme linked immunosorbent assay (i-ELISA) and Western Blot (WB) tests. The eighth collection was made six months after the seventh, using the WB and Polymerase Chain Reaction (PCR). The prevalence of CAE was 6.8%, 14.9% and 39.2% in the AGID, i-ELISA and WB respectively. The last analysis detected 0.9% of animals positive by WB and 10.8% by PCR. Although they do not eradicate CAE, steps taken together with the periodic use of serological tests and the combination with PCR were important to significantly reduce positive animals in the herd.(AU)

In vitro and in vivo evaluation of sodium dodecyl sulfate (SDS) as an inactivator of caprine lentivirus (CLV) in colostrum and milk / Avaliação in vitro e in vivo do dodecil sulfato de sódio (SDS) como inativador do lentivírus caprino (LVC) em colostro e leite

The aim of this study was to evaluate in vitro and in vivo the effect of sodium dodecyl sulfate (SDS) on the caprine lentivirus (CLV) in colostrum and milk. This was performed to develop a practical and efficient method of blocking the lactogenic transmission of the virus. In the in vitro experiment, colostrum and milk were treated with 0.25%; 0.50% and 1% SDS. Then, somatic cells of colostrum and milk were submitted to co-culture with caprine synovial membrane cells (CSM). In the in vivo test, goats were fed with colostrum and milk provided from CLV-positive goats treated with SDS in the same concentrations used in the in vitro experiment. Animals were tested by nested polymerase chain reaction (nPCR) and Western blot (WB) assays. In the in vitro experiment, inhibitory activity against CLV without inactivation occurred in colostrum with all SDS concentrations. However, concentrations of 0.25 and 0.5% SDS presented only inhibitory activity against CLV in milk cells, and 1% concentration provided inactivation of the virus. In the in vivo tests, none of the three concentrations of SDS was effective in inactivating LVC in colostrum or goat milk, which was confirmed by seroconversion and presence of proviral DNA in animals afterwards.(AU)

O objetivo da pesquisa foi avaliar in vitro e in vivo o efeito do dodecil sulfato de sódio (SDS) sobre o lentivírus caprino (LVC) no colostro e no leite, a fim de desenvolver um método prático e eficiente no bloqueio da via de transmissão lactogênica do vírus. No experimento in vitro, o colostro e o leite de cabras positivas foram tratados com SDS a 0,25%, 0,50% e 1,0%. Em seguida, as células somáticas do colostro e do leite foram obtidas e direcionadas ao cocultivo com células de membrana sinovial caprina (MSC). No teste in vivo, os cabritos foram alimentados com colostro e leite providos de cabras positivas para LVC, tratados com SDS nas mesmas concentrações usadas no teste in vitro. Os animais foram acompanhados pelos testes de reação em cadeia da polimerase nested (nPCR) e western blot (WB). Nos resultados in vitro, no colostro, observou-se que, em todas as concentrações de SDS, ocorreu uma atividade inibitória contra o LVC, sem a inativação. Em relação às células do leite, o SDS apresentou, nas concentrações de 0,25 e 0,5%, atividade inibitória contra o LVC, e na concentração de 1%, houve inativação viral. Nos testes in vivo, as três concentrações de SDS testadas não foram efetivas na inativação do LVC no colostro e no leite caprino, o que se comprovou pela soroconversão e pela presença de DNA proviral nos animais.(AU)

Wharton's jelly cells from sheep umbilical cord maintained with different culture media are permissive to in vitro infection by Small Ruminant Lentiviruses / Células da geleia de Wharton do cordão umbilical ovino mantidas em diferentes meios de cultivo são permissivas à infecção in vitro por lentivírus de pequenos ruminantes

This study aimed to isolate cells from the Wharton's jelly of umbilical cord (WJUC) of sheep collected during natural parturition using different culture media, in addition to reporting for the first time the permissiveness of these cells to in vitro infection by small ruminant lentiviruses. Ten umbilical cords were collected from healthy sheep. Each cord explants were grown in different media consisting of MEM, low glucose DMEM, M199, and RPMI-1640. The permissiveness of infection of sheep cells from WJUC was tested with CAEV-Cork and MVV-K1514 strains, inoculating 0.1 MOI of each viral strain. Four supernatants from each strain were obtained from WJUC sheep cell cultures infected in different media. The results demonstrated the presence of cytopathic effect after the in vitro infection by CAEV-Cork and MVV-K1514 with all of the tested culture media. Nested-PCR detected proviral DNA in all supernatants. Supernatants containing CAEV-Cork viruses had TCID 50/ml titres of 10 5.5 in MEM, 10 4.0 in low glucose DMEM, 105.0 in M199, and 10 5.7 in RPMI-1640. Supernatants containing the MVV-K1514 virus had TCID 50/ml titres of 10 4.3 in MEM, 10 3.5 in low-glucose DMEM, 10 4.7 in M199, and 10 3.5 in RPMI-1640. Sheep cells from WJUC are permissive to in vitro infection by small ruminant lentivirus.(AU)

O objetivo deste estudo foi isolar células da geleia de Wharton do cordão umbilical (GWCU) ovino coletado por ocasião do parto natural, utilizando-se diferentes meios de cultivo, além de relatar, pela primeira vez, sua permissividade à infecção in vitro por lentivírus de pequenos ruminantes (LVPRs). Dez cordões umbilicais foram coletados de ovelhas hígidas e soronegativas para LVPRs pelo teste de imunodifusão em gel de agarose (IDGA). De cada cordão, explantes foram cultivados em quatro meios distintos que consistiram em MEM, DMEM baixa glicose, meio 199 e RPMI-1640, todos acrescidos de 10% de soro fetal bovino em estufa com atmosfera úmida e 5% de CO2 a 37ºC. A permissividade de infecção das células GWCU ovino foi testada frente às cepas CAEV-Cork e MVV-K1514, inoculando-se 0,1 MOI de cada cepa viral e corando as monocamadas com May Grunwald Giemsa para visualização do efeito citopático. Foram obtidos quatro sobrenadantes CAEV-Cork e quatro MVV-K1514, provenientes do cultivo de células GWCU ovino infectadas por 21 dias em meios distintos, dos quais foram realizadas titulação em membrana sinovial caprina e extração do DNA pró-viral para realização de nested-PCR e eletroforese em gel de agarose a 2%. Os resultados demonstraram a presença de efeito citopático na infecção in vitro tanto por CAEV-Cork como por MVV-K1514 em todos os meios de cultivo, sendo visualizados sincícios e lise celular em microscópio invertido. A nested-PCR detectou o DNA pró-viral tanto do CAEV-Cork como do MVV-K1514 em todos os sobrenadantes. Os sobrenadantes contendo o vírus CAEV-Cork apresentaram títulos em TCID50/mL de 10 5,5 em MEM, 10 4,0 em DMEM baixa glicose, 10 5,0 em meio 199 e 10 5,7 em RPMI-1640. Os sobrenadantes contendo o vírus MVV-K1514 apresentaram título em TCID 50/mL de 10 4,3 em MEM, 10 3,5 em DMEM baixa glicose, 10 4,7 em meio 199 e 10 3,5 em RPMI-1640. Células GWCU ovino são permissivas à infecção in vitro pelos lentivírus de pequenos ruminantes CAEV-Cork e MVV-K1514.(AU)

Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina / Standardization of indirect Elisa and Western Blot for diagnosis of Caprine Arthritis-Encephalitis

A artrite-encefalite caprina (CAE) é diagnosticada rotineiramente pela técnica de imunodifusão em gel de agarose (IDGA), que é considerada pouco sensível. Objetivou-se com este estudo padronizar testes de Elisa-i e Western Blot (WB) para diagnóstico precoce de anticorpos em caprinos contra CAEV e comparar os resultados obtidos nesses testes com a prova de IDGA. Para a padronização dos testes Elisa-i e WB, utilizaram-se diferentes concentrações e diluições de antígeno, soros e conjugado. No Elisa-i, adotaram-se microplacas rígidas com 96 poços, sendo a combinação de concentração de 0,5µg/poço de antígeno e diluições de 1:100 de soro e 1:1500 de conjugado a que apresentou melhor resultado. No WB foram utilizadas membranas de nitrocelulose, definindo-se as diluições de 1:50 de soro e 1:15000 de conjugado. Para avaliar o desempenho das técnicas, 222 amostras de soro caprino foram testadas e os dados obtidos foram comparados com o IDGA. A sensibilidade e a especificidade do Elisa-i/IDGA, WB/IDGA e WB/Elisa-i foram de 70% e 91%, 100% e 72,6%, 84,6% e 76,5%, concomitantemente. O índice Kappa desses testes foi de 0,35, 0,2 e 0,36, respectivamente. As técnicas de Elisa-i e WB apresentaram-se mais sensíveis que a IDGA, podendo ser utilizadas como ferramentas para o diagnóstico precoce da CAE...

Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE...

Produção de biocompostos com atividade antimicrobiana de Streptomyces sp. ante isolados de mastite caprina / Production of biocompounds with antimicrobial activity of Streptomyces sp. Isolated in face of goat mastitis

Actinomicetos são um dos principais produtores de enzimas, vitaminas e metabólitos secundários, destacando-se o gênero Streptomyces, o qual tem uma ampla capacidade de produção de antibióticos eficazes no combate a diferentes microrganismos, entre eles o Staphylococcus sp. Em virtude dessa eficiência no combate a patógenos, o objetivo deste trabalho foi avaliar a produção de metabólitos com atividade antimicrobiana produzidos por 67 Streptomyces isolados de liquens da região amazônica, ante isolados de mastite caprina (Staphylococcus aureus) do estado de Pernambuco, Brasil. Foi utilizado um planejamento fatorial para avaliar a influência das fontes de carbono (glicose) 0%, 0,5% e 1% e de nitrogênio (farinha de soja) 1%, 2,5% e 4% na produção dos antimicrobianos, bem como das variáveis pH, biomassa e atividade antimicrobiana. Dos Streptomyces estudados, o DPUA 1566 foi o que se destacou por formação de halos de inibição entre 18 e 26mm ante os isolados de mastite caprina. Foi possível verificar que a fonte de carbono inibiu a produção de antimicrobianos quando submetidos a uma concentração de glicose de 1%; com a retirada desta, os Streptomyces apresentaram uma elevada capacidade de produção de metabólitos com atividade antimicrobiana tendo potencial para o tratamento de mastite caprina.

Actinomycetes are a leading producer of enzymes, vitamins and secondary metabolites, especially the genus Streptomyces, which have a large capacity for the production of natural antibiotics and are effective against various micro-organisms including Staphylococcus sp. Due to this efficiency in combating micro-organisms, the aim of this study was to evaluate the production of metabolites with antimicrobial activity produced by Streptomyces isolated from lichens 67 in the Amazonia, compared to isolates from goat mastitis (Staphylococcus aureus) in the state of Pernambuco, Brazil. We used a complete factorial design to evaluate the influence of concentrations of carbon sources (glucose) at 0%, 0.5% and 1% and nitrogen (soybean flour) at 1%, 2.5% and 4% in the production of antimicrobial metabolites, and the influence of the pH, microbial biomass and activity variables. Of the studied Streptomyces DPUA 1566 what stood out was the formation of inhibition halos between 18 to 26 mm compared to the isolates from goats mastitis. It was noted that the carbon source inhibited the production of antimicrobial metabolites when subjected to a glucose concentration of 1%. However, after the discontinuation, Streptomyces showed a high capacity to produce metabolites with antimicrobial activity, which has an excellent potential for the treatment of mastitis in goats.

Biomechanical and structural parameters of tendons in rats subjected to swimming exercise.

The aim of this study was to evaluate the effect of swimming exercise, without overloading, on the biomechanical parameters of the calcaneal tendon of rats. 27 male Wistar rats (70 days) were distributed randomly into 2 groups, Control Group (CG; n=15) with restricted movements inside the cage and Swimming Group (SG; n=12), subjected to exercise training in a tank with a water temperature of 30±1°C, for 1 h/day, 5 days/week for 8 weeks. All animals were kept in a reversed light/dark cycle of 12 h with access to food and water ad libitum. After that, they were anesthetized and had their calcaneus tendons collected from their left rear paws. The tendon was submitted to a mechanical test on a conventional test machine. From the stress vs. strain curve, the biomechanical data were analyzed. For the statistical analysis, the Student-T test was used (p<0.05). Of the variables examined, the maximum tension (p=0.009), maximum force (p=0.03), energy of deformation/tendon cross sectional area (p=0.017) and elastic modulus of the tendon (p=0.013) showed positive outcomes in SG. There was no difference in the other parameters. The results indicate that the swimming exercise training, without overloading, was an important stimulus for improving the biomechanical parameters and structural properties of the calcaneal tendon.

Frequência de anticorpos contra o vírus da língua azul em ovinos do estado do Ceará, Brasil / Antibodies against the bluetongue virus in sheep flocks of Ceará state, Brazil

O objetivo deste trabalho foi avaliar a ocorrência de ovinos soropositivos para o vírus da línguaazul (VLA) no Estado do Ceará, Brasil, e analisar as proteínas imunogênicas das cepas virais circulantes nesses rebanhos. O teste de imunodifusão em gel de agarose (IDGA) foi utilizado para pesquisar 271 amostras de soro oriundas de 16 rebanhos. Os resultados demonstraram que 27,3% (74/271) das amostras analisadas apresentaram anticorpos contra o agente e 68,8% (11/16) das propriedades tiveram animais positivos. O immunoblotting (IB) foi utilizado para analisar as proteínas imunogênicas do VLA a partir dos soros de animais positivos no IDGA. Os soros demonstraram forte reação contra a proteína viral VP2. Para o VLA, das sete proteínas estruturais, a VP2 é a principal a estimular a resposta imune protetora. Concluiu-se que a soropositividade para a língua azul (LA) nos rebanhos ovinos estudados no Ceará é alta, apesar dos animais não apresentarem sinais clínicos, indicativo de que o vírus ocorra de forma endêmica. Além disso, a resistência à doença apresentada pelos animais pode estar relacionada com a forte reação imunológica desses à proteína VP2. Sendo assim, outros estudos são necessários para melhor esclarecer a situação epidemiológica da LA no país, através da identificação dos vetores e sorotipos virais circulantes nas diferentes regiões.

Antibodies against the bluetongue virus in sheep flocks of Ceará state, Brazil. The objective of this work was to verify the occurrence of sheep serologically positive for bluetongue virus (BTV) in the state of Ceará, Brazil, and analyze immunogenic proteins of circulating viral strains in these flocks. The agar gel immunodifusion test (AGID) was used to examine 271 serum samples from 16 herds. The results demonstrated that 27.3% (74/271) ofthe analyzed samples presented antibodies for the agent, and that 68.8% (11/16) of the propertiespresented positive animals. Immunoblotting (IB) was used to analyze the immunogenicproteins of BTV derived from AGID positive sera. Sera showed strong reaction against viral protein VP2. Of the seven BTV structural proteins, VP2 is the major protein to elicit protective immuneresponses. It was concluded that bluetongue (BT) seropositivity in sheep flocks studied in Ceará is high, despite that the animal's do not show clinical signs, indicating that it occurs in an endemic form. The animals’ resistance to the disease may be related to the strong immune response to the protein VP2. Therefore, further studies are needed to better clarify the epidemiological situation of BT in Brazilian sheep flocks, through the identification of viral vectors and serotypes circulating in different regions.

Alterações histopatológicas da glândula mamária e qualidade do leite de cabras naturalmente infectadas com o CAEV / Histopathological changes in the mammary gland and milk quality of goats naturally infected with CAEV

Avaliou-se a influência do vírus da CAE nas características físico-químicas de amostras de leite de 54 cabras, sem predileção racial, distribuindo-as em dois grupos: cabras positivas e negativas para o teste de imunodifusão em gel de agarose. As amostras de leite foram submetidas à análise ultrassônica para obtenção de parâmetros físico-químicos - gordura, extrato seco, proteínas, lactose e densidade; realização de microbiologia - bactérias mesófilas (UCF/mL). Foram coletadas amostras de tecido mamário para exame histopatológico e imunohistoquímica. Não houve diferença significativa das características avaliadas entre os dois grupos; no microbiológico, não houve relação direta da presença de mesófilas associada à infecção pelo CAEV. Na histopatologia, observaram-se áreas com infiltração celular de monócitos, polimorfonucleares, plasmócitos, fibrose, ausência de morfologia normal do parênquima mamário, denotando processo inflamatório crônico; e foi confirmada a presença do vírus na glândula pela imunohistoquímica. Os resultados não mostraram relação direta da incidência da CAE como fator negativo no desenvolvimento do rebanho.

Aiming to evaluate the influence of CAE viruses in the chemical and physical characteristics of milk, the samples were collected from 54 goats, without racial predilection, these were divided into two groups: goats positive and negative according results of test Agarose Gel Immunodiffusion. Milk samples were ultrasonic analyzed to obtain physicochemical parameters (fat, solids, protein, lactose and density); performance microbiology (mesophilic bacteria - CFU/mL) and mammary gland samples were collected for evaluation histopathology and immunohistochemistry. The results of physical-chemical analysis showed no significant difference between the milk samples of two groups. In the microbiological analysis showed the presence of aerobic mesophilic bacteria, but this change is not associated with the presence of CAEV infection. On histopathology, there were areas with infiltration of mononuclear-leukocyte and polymorph nuclear, plasma cells, fibrosis and absence of normal morphology of the mammary tissue, indicating a chronic inflammatory process; and confirmed the presence of virus, in the gland, by immunohistochemistry. The results showed no direct relationship between incidence of CAE in the herd as a negative factor for its development, however it is known that the disease in its chronic nature, causes reduction in the productivity of the herd.

Produção de antígeno e separação da proteína p28 por microfiltragem seriada para sorodiagnóstico da artrite encefalite caprina por ensaio imunoenzimático / Production of antigen and p28 protein separation by microfiltration serial for serodiagnosis of caprine arthritis encephalitis by enzyme immunoassay

Este estudo teve como objetivo produzir um antígeno (Ag) a partir de cultura de células de membrana sinovial caprina (MSC) infectadas com o vírus de artrite encefalite caprina (CAEV), pela técnica de microfiltração seriada, substituindo a ultracentrifugação em colchão de sacarose (UCCS) para utilização em ELISA indireto (ELISA-i). Amostras de 188 soros caprinos, que previamente foram testados pelo Western blot (WB) com Ag UCCS, foram submetidas à análise pelo ELISA-i com o novo antígeno produzido, que mostrou concordância de 92% em relação ao antígeno UCCS. A sensibilidade e a especificidade do ELISA em relação ao WB foram de 95,6% e 88,5%, respectivamente. A nova técnica, criada a partir de microfiltrações, mostrou-se efetiva e de baixo custo para o diagnóstico sorológico de anticorpos para CAEV em comparação ao antígeno ultracentrifugado, e constitui uma alternativa viável para produção de antígeno purificado de lentivírus de pequenos ruminantes.

This study aimed to produce an antigen (Ag) from the culture of goat synovial membrane cells (MSC) infected by CAEV through serial microfiltering technique replacing ultra ultracentrifugation in sacarosis Mattress (UCCS) for the indirect diagnosis ELISA tests (i ELISA). Samples of 188 sera from goats previously examined by Western Blot (WB) with Ag UCCS were submitted to analysis by i ELISA with new antigen produced, demonstrating an accordance of 92% in relation to UCCS antigen. The specificity and sensitivity relating to WB were of 95,65% and 88, 5% respectively. The new technique created from the microfiltering is effective and with low cost for the serological antibodies diagnosis of CAEV comparing to the ultracentrifuged one, presenting, therefore, as a viable alternative for purified antigen of lentivirus in small ruminants.

Actinomicetos produtores de inibidores de beta-lactamases com atividade antimicrobiana frente a isolados de mastite bovina / Actinomycetes producers of beta-lactamases inhibitors with antimicrobial activity against microorganisms isolated from bovine mastitis

Avaliou-se a capacidade de 71 actinomicetos isolados de líquens da região amazônica em produzir inibidores de β-lactamases com atividade antimicrobiana sobre Staphylococcus aureus, resistentes à penicilina, isolados de mastite bovina do estado de Pernambuco. A seleção dos actinomicetos produtores de inibidores de β-lactamases foi realizada pela técnica de bloco de gelose contra Klebsiella pneumoniae ATCC 29665, e os actinomicetos selecionados foram testados frente a 17 linhagens de Staphylococcus aureus resistentes à penicilina. Os melhores produtores de inibidores de β-lactamases foram Streptomyces sp. DPUA 1542 e Nocardia sp. DPUA 1571, os quais foram submetidos ao cultivo submerso para determinação da curva de crescimento, pH e atividade antimicrobiana. Os maiores halos de inibição foram obtidos pelos metabólitos produzidos após 96 horas de cultivo tanto para Nocardia sp. - 13,5 e 12,0mm - como para Streptomyces sp. - 8,0 e 14,0mm - com os testes de difusão nos discos e poços, respectivamente. Os resultados permitiram concluir que os actinomicetos são fonte promissora de inibidores de β-lactamases, com potencial uso no tratamento de mastites bovinas.

The ability of 71 actinomycetes, isolated from the Amazon lichens, to produce β-lactamase inhibitors with antimicrobial activity was evaluated against penicillin-resistant Staphylococcus aureus, isolated from bovine mastitis in Pernambuco State. The selection of actinomycetes producers of β-lactamase inhibitors was performed using agar-plug method against Klebsiella pneumoniae ATCC 29665 and the selected actinomycetes were tested against 17 penicillin-resistant Staphylococcus aureus strains. The best producers of β-lactamase inhibitors were Streptomyces sp. DPUA 1542 and Nocardia sp. DPUA 1571. They were submitted to the submerged cultivation to determine the growth and pH curve, and antimicrobial activity. The highest inhibition halo zonewas obtained by metabolites produced after 96 hours of cultivation for both Nocardia sp. (13.5 and 12.0mm) and Streptomyces sp. (8.0 and 14.0mm) with discs and well diffusion tests, respectively. The results showed that the actinomycetes are a promising source of β-lactamase inhibitors, with potential for use in the bovine mastitis treatment.

Avaliação imunohistoquímica e ultraestrutural de gametas e embriões caprinos infectados com o CAEV / Immunohistochemical and ultrastructural evaluation of gametes and embryos of goats infected with CAEV

O objetivo do presente estudo foi determinar a susceptibilidade dos folículos ovarianos, espermatozoides e embriões caprinos ao Vírus da Artrite Encefalite Caprina (CAEV). Para isto, foram analisados espermatozoides e folículos ovarianos pelas técnicas de imunohistoquímica e microscopia eletrônica de transmissão, antes e após protocolos de infecção in vitro com o CAEV. Foram submetidos à análise ultraestrutural, embriões caprinos produzidos in vivo, oriundos de cabras negativas e positivas para o CAEV. Nas amostras seminais, provenientes de animais tanto com infecção natural quanto dos artificialmente infectados, foi observada imunomarcação positiva dos espermatozoides, assim como alterações degenerativas na sua análise ultraestrutural. Já nas amostras de tecido ovariano, a imunomarcação foi mais discreta e identificada na região do estroma. No tocante à análise ultraestrutural, folículos e embriões se apresentaram íntegros. De acordo com esses resultados, pode-se concluir que os espermatozoides caprinos apresentaramse infectados, assinalando a susceptibilidade dessas células ao vírus, bem como a potencialidade do CAEV ser carreado ao cerne do oócito, originando embriões infectados.

The aim of this study was to determine the susceptibility of goat ovarian follicles, spermatozoa and embryos to caprine arthritis-encephalitis virus (CAEV). Spermatozoa and ovarian follicles were analyzed, before and after in vitro infection with CAEV, through immunohistochemistry and transmission electron microscopy techniques. Goat embryos, produced in vivo from infected and non-infected goats, were submitted to ultrastructural analysis. Immunohistochemical examination of seminal samples from goats naturally and artificially infected with CAEV revealed viral antigens in spermatozoa, while the ultrastructural analysis showed degenerative changes in these cells. Ovarian tissue samples presented a more discreet immunohistochemical positive reaction situated in the stroma region. Ultrastructural analysis revealed that the embryos and ovarian follicles were intact. These results indicate that the spermatozoa were infected, confirming the susceptibility of these cells to the virus, as well as the potential of CAEV entering the oocyte, giving rise to infected embryos.

Inibição dos lentivírus de pequenos ruminantes por drogas antivirais / Inhibition of small ruminant lentivirus by antiviral drugs

Inibidores da enzima transcriptase reversa e da protease foram avaliados quanto ao seu efeito inibitório na replicação do Vírus da Artrite Encefalite Caprina (CAEV) cepa CAEV Cork e do vírus Maedi-Visna (MVV) cepa K1514 cultivados em células fibroblásticas de caprinos. Os fármacos utilizados foram: lamivudina, didanosina, estavudina, zidovudina, efavirenz, atazanavir e lopinavir/ritonavir. A maior concentração utilizada para lamivudina, estavudina, zidovudina e efavirenz foi 500 ?M, para atazanavir foi 50 ?M e 5,0 ?M para lopinavir/r e didanosina. A atividade antiviral in vitro foi pesquisada por meio da avaliação da viabilidade celular através da redução do MTT e pela pesquisa de inibição dos efeitos citopáticos (CPE) dos vírus. A replicação dos vírus só não foi completamente bloqueada pelos inibidores de protease (IP) atazanavir e lopinavir/r enquanto os demais apresentaram eficácia antiviral significativa em diferentes concentrações. Os IP juntamente com o efavirenz, não mostraram atividade antiviral quando foram avaliados pela técnica de redução do MTT. Esses dados indicam que os fármacos inibidores da transcriptase reversa lamivudina, didanosina, estavudina e zidovudina são eficazes na inibição in vitro dos lentivírus de pequenos ruminantes.

Inhibitors of the reverse transcriptase and protease enzymes were evaluated for their inhibitory effect on the replication of caprine arthritis encephalitis virus (CAEV) strain CAEV Cork and of the Maedi-Visna virus (MVV) strain K1514 cultured in fibroblastic cells. The drugs lamivudine, didanosine, stavudine, zidovudine, efavirenz, atazanavir and lopinavir/ritonavir were used. The highest concentration used for lamivudine, stavudine, zidovudine and efavirenz was 500 ?M, for atazanavir it was 50 ?M and 5.0 ?M for lopinavir/r and didanosine. The in vitro antiviral activity was investigated by evaluating the cell viability by the MTT method and testing for inhibition of cytopathic effects (CPE) of the virus. The replication of the virus was not completely inhibited by the protease inhibitors atazanavir and lopinavir/r in the test for CPE, while the others drugs showed significant antiviral efficacy in different concentrations. The protease inhibitors together with the efavirenz did not show antiviral activity when they were assessed by the reduced MTT technique. These data showed that the reverse transcriptase inhibitor drugs lamivudine, didanosine, stavudine and zidovudine were effective in the in vitro inhibition of small ruminant lentivirus.

Temporal trends in incidence of Type 1 diabetes between 1986 and 2006 in Brazil.

BACKGROUND: Scarce information is available about the variation in the incidence of Type 1 diabetes in the Brazilian population in the last decades. AIM: The objective of this study was to assess the long-term trends (1986-2006) in the incidence of Type 1 diabetes in Bauru, São Paulo State, Brazil. SUBJECTS AND METHODS: The annual incidence of Type 1 diabetes (per 100,000 per yr) from 1986 to 2006 was determined in children or=20/100,000 per yr) in 71.43% of the study-years. Incidence was slightly higher among females, Caucasians, children in the 5-9 yr of age range and belonging to lower socio-economic classes. Most diagnoses were established during the colder months and/or with higher pluviometric indexes. CONCLUSIONS: The incidence of Type 1 diabetes in children is increasing in Bauru, São Paulo State, Brazil, and the global pattern of incidence was classified as high or very high, mainly in the last 10 yr. All Brazilian regions should be involved in the study.

Phenotypic characterization of three clinical isolates of Burkholderia pseudomallei in Ceará, Brazil.

Burkholderia pseudomallei, the causative agent of melioidosis was found in a small cluster of cases in Tejuçuoca, Ceará, Brazil. Tests were carried out to determine its phenotypic characteristics: colony morphology on Ashdown agar and MacConkey agar, biochemical profile in conventional biochemical tests and API 20NE, arabinose assimilation and susceptibility testing by disk diffusion, comparing with data in the literature. This study confirms the presence of B. pseudomallei in Brazil and describes its characteristics.

Phenotypic characterization of three clinical isolates of Burkholderia pseudomallei in Ceará, Brazil

Burkholderia pseudomallei, the causative agent of melioidosis was found in a small cluster of cases in Tejuçuoca, Ceará, Brazil. Tests were carried out to determine its phenotypic characteristics: colony morphology on Ashdown agar and MacConkey agar, biochemical profile in conventional biochemical tests and API 20NE, arabinose assimilation and susceptibility testing by disk diffusion, comparing with data in the literature. This study confirms the presence of B. pseudomallei in Brazil and describes its characteristics.

High frequencies of plexiform neurofibromas, mental retardation, learning difficulties, and scoliosis in Brazilian patients with neurofibromatosis type 1.

A clinical study of Brazilian patients with neurofibromatosis type 1 (NF1) was performed in a multidisciplinary Neurofibromatosis Program called CEPAN (Center of Research and Service in Neurofibromatosis). Among 55 patients (60% females, 40% males) who met the NIH criteria for the diagnosis of NF1, 98% had more than six café-au-lait patches, 94.5% had axillary freckling, 45% had inguinal freckling, and 87.5% had Lisch nodules. Cutaneous neurofibromas were observed in 96%, and 40% presented plexiform neurofibromas. A positive family history of NF1 was found in 60%, and mental retardation occurred in 35%. Some degree of scoliosis was noted in 49%, 51% had macrocephaly, 40% had short stature, 76% had learning difficulties, and 2% had optic gliomas. Unexpectedly high frequencies of plexiform neurofibromas, mental retardation, learning difficulties, and scoliosis were observed, probably reflecting the detailed clinical analysis methods adopted by the Neurofibromatosis Program. These same patients were screened for mutations in the GAP-related domain/GRD (exons 20-27a) by single-strand conformation polymorphism. Four different mutations (Q1189X, 3525-3526delAA, E1356G, c.4111-1G>A) and four polymorphisms (c.3315-27G>A, V1146I, V1317A, c.4514+11C>G) were identified. These data were recently published.