Anticorpos contra vírus do grupo da língua azul em caprinos e ovinos do sertão de Pernambuco e inferências sobre sua epidemiologia em regiões semiáridas / Antibodies against bluetongue-virus group in goats and sheep from Pernambuco state and inferences on bluetongue epidemiology under tropical conditions

The prevalence of antibodies against bluetongue virus was investigated in 41 dairy goats and 40 sheep herds in the semi-arid region of Pernambuco state and the conditions for insect Culicoides maintenance, considering climate dynamics and vector competence, were evaluated. The percents of seropositive herds in agar gel immunodiffusion test for bluetongue virus group were 24 for goats and 27.5 for sheep. The estimated prevalences of seropositive animals were 3.9 percent for goats (n = 410) and 4.3 percent for sheep (n = 400). The prevalences of seropositive animals were low in the mesoregion of Sertão Pernambucano (4.8 percent for goats and 4.1 percent for sheep) and São Francisco Pernambucano (1.0 percent for goats and 4.5 percent for sheep). There were no significant differences between species and regions. Considering the social and economic importance of goats and sheep raising in the semi-arid region, it is essential to establish preventive measures to control imports of ruminants from these areas.

Anticorpos contra vírus do grupo da língua azul em caprinos e ovinos do sertão de Pernambuco e inferências sobre sua epidemiologia em regiões semiáridas / Antibodies against bluetongue-virus group in goats and sheep from Pernambuco state and inferences on bluetongue epidemiology under tropical conditions

The prevalence of antibodies against bluetongue virus was investigated in 41 dairy goats and 40 sheep herds in the semi-arid region of Pernambuco state and the conditions for insect Culicoides maintenance, considering climate dynamics and vector competence, were evaluated. The percents of seropositive herds in agar gel immunodiffusion test for bluetongue virus group were 24 for goats and 27.5 for sheep. The estimated prevalences of seropositive animals were 3.9 percent for goats (n = 410) and 4.3 percent for sheep (n = 400). The prevalences of seropositive animals were low in the mesoregion of Sertão Pernambucano (4.8 percent for goats and 4.1 percent for sheep) and São Francisco Pernambucano (1.0 percent for goats and 4.5 percent for sheep). There were no significant differences between species and regions. Considering the social and economic importance of goats and sheep raising in the semi-arid region, it is essential to establish preventive measures to control imports of ruminants from these areas.(AU)

FREQUÊNCIA DE ANTICORPOS E FATORES DE RISCO ASSOCIADOS À LEUCOSE ENZOÓTICA BOVINA EM REBANHOS DA BACIA LEITEIRA DO ESTADO DO MARANHÃO

ABSTRACT This study was performed in order to determine the frequency of antibodies to enzootic bovine leukosis (EBL), and the main factors related to the infection in dairy herds of Maranhão State, Brazil. Serum samples from 920 animals were submitted to the agar gel immunodifusion technique. The study was carried out on 92 dairy properties, in 23 districts located in the regions of Açailândia, Bacabal, São Luis Island, Imperatriz and Pedreiras. An epidemiological questionnaire was applied for each herd to investigate variables that could be associated with this infection. Of the 920 serum samples examined, 53.8% were reagent. The frequency breakdown by region was 63.50%, 61.87%, 60.62%, 41.18% and 30.83%, for Bacabal, São Luis Island, Pedreiras, Imperatriz and Açailândia, respectively. Reagents animals were found in the 23 districts studied, with detection of serologically positive bovines on 98.91% of the properties. Variables identified as risk factors included the repeated use of the same needle for blood sampling or vaccination, repeated use of the same obstetric glove, animal housing and absence of veterinary assistance. These risk factors presented a statistical significance (P 0.05) associated to infection. These results indicate that enzootic bovine leukosis infection is high in dairy herds of the state of Maranhão and that factors related to livestock management were associated with the risk of EBL infection.

RESUMO Este trabalho teve como objetivo determinar a frequência e os possíveis fatores de risco as-sociados à ocorrência da Leucose Enzoótica Bovina (LEB) na bacia leiteira do Estado do Maranhão. Amostras de soro de 920 animais da raça girolanda foram submetidas à técnica de imunodifusão em gel de ágar. O estudo foi realizado em 92 propriedades leiteiras, pertencentes a 23 municípios localizados nas regionais de Açailândia, Bacabal, Ilha de São Luís, Imperatriz e Pedreiras. Em cada propriedade avaliada, aplicou-se questionário epidemiológico para investigar os fatores de risco que poderiam estar associados a esta infecção. Das 920 amostras de soro analisadas, 53,80% foram reagentes. Nas regionais obtiveram-se frequências de 63,50%, 61,87%, 60,62%, 41,18% e 30,83%, para Bacabal, Ilha de São Luís, Pedreiras, Imperatriz e Açailândia, respectivamente. Nos 23 municípios amostrados foram encontrados animais reagentes, com detecção de bovinos sorologicamente positivos em 98,91% das propriedades. Dentre os fatores de risco avaliados, uso repetido da mesma agulha para colheita de sangue ou vacinação, uso repetido da mesma luva obstétrica, estabulação dos animais e ausência de assistência veterinária apresentaram significância estatística (P 0,05) associada à LEB. Os resultados do estudo indicam que a frequência de anticorpos para a Leucose Enzoótica Bovina foi elevada na bacia leiteira do Estado do Maranhão e que os fatores relacionados ao manejo estiveram associados ao risco de infecção nos bovinos.

Frequência de anticorpos e fatores de risco associados à leucose enzoótica bovina em rebanhos da bacia leiteira do estado do Maranhão / Frequency of antibodies and risk factors associated to enzootic bovine leukosis in dairy herds of the state of Maranhão, Brazil

Este trabalho teve como objetivo determinar a frequência e os possíveis fatores de risco as-sociados à ocorrência da Leucose Enzoótica Bovina (LEB) na bacia leiteira do Estado do Maranhão. Amostras de soro de 920 animais da raça girolanda foram submetidas à técnica de imunodifusão em gel de ágar. O estudo foi realizado em 92 propriedades leiteiras, pertencentes a 23 municípios localizados nas regionais de Açailândia, Bacabal, Ilha de São Luís, Imperatriz e Pedreiras. Em cada propriedade avaliada, aplicou-se questionário epidemiológico para investigar os fatores de risco que poderiam estar associados a esta infecção. Das 920 amostras de soro analisadas, 53,80% foram reagentes. Nas regionais obtiveram-se frequências de 63,50%, 61,87%, 60,62%, 41,18% e 30,83%, para Bacabal, Ilha de São Luís, Pedreiras, Imperatriz e Açailândia, respectivamente. Nos 23 municípios amostrados foram encontrados animais reagentes, com detecção de bovinos sorologicamente positivos em 98,91% das propriedades. Dentre os fatores de risco avaliados, uso repetido da mesma agulha para colheita de sangue ou vacinação, uso repetido da mesma luva obstétrica, estabulação dos animais e ausência de assistência veterinária apresentaram significância estatística (P < 0,05) associada à LEB. Os resultados do estudo indicam que a frequência de anticorpos para a Leucose Enzoótica Bovina foi elevada na bacia leiteira do Estado do Maranhão e que os fatores relacionados ao manejo estiveram associados ao risco de infecção nos bovinos.

This study was performed in order to determine the frequency of antibodies to enzootic bovine leukosis (EBL), and the main factors related to the infection in dairy herds of Maranhão State, Brazil. Serum samples from 920 animals were submitted to the agar gel immunodifusion technique. The study was carried out on 92 dairy properties, in 23 districts located in the regions of Açailândia, Bacabal, São Luis Island, Imperatriz and Pedreiras. An epidemiological questionnaire was applied for each herd to investigate variables that could be associated with this infection. Of the 920 serum samples examined, 53.8% were reagent. The frequency breakdown by region was 63.50%, 61.87%, 60.62%, 41.18% and 30.83%, for Bacabal, São Luis Island, Pedreiras, Imperatriz and Açailândia, respectively. Reagents animals were found in the 23 districts studied, with detection of serologically positive bovines on 98.91% of the properties. Variables identified as risk factors included the repeated use of the same needle for blood sampling or vaccination, repeated use of the same obstetric glove, animal housing and absence of veterinary assistance. These risk factors presented a statistical significance (P < 0.05) associated to infection. These results indicate that enzootic bovine leukosis infection is high in dairy herds of the state of Maranhão and that factors related to livestock management were associated with the risk of EBL infection.

Western blot para o diagnóstico das infecções pelos lentivírus de pequenos ruminantes em caprinos: um método simples para a produção de antígeno

ABSTRACT The serodiagnosis of small ruminant lentivirus (SRLV) infections in goats and sheep is usually performed by the agar-gel immunodiffusion technique (AGID) and by ELISA. Therefore, the western blot (WB) is the choice for the definition of antigen recognition patterns during the disease progression, being potentially useful for the diagnosis itself. In the present study, the antigen used for WB was obtained by a novel simple purification procedure: the dialysis of the supernatant of infected cultured cells, followed by centrifugation in sucrose gradient. Proteins in the pellet were separated by SDS-PAGE at 10% density and transferred to nitrocellulose membranes in a semi-dry blotting device. After development, five viral proteins were recognized by the standard positive serum sample, with molecular weights of 14-16, 25, 40, 50 and 70 kDa respectively. All 8 AGID positive goat serum samples recognized at least one band in the immunoblot, with different intensities. The total number of bands recognized by each serum sample varied considerably. A positive reaction to gp40 was observed with 4 sera, although rather weak in 3 cases. Two samples were reactive to p16 and two others to gp50, although weakly. Curiously, no serum was reactive to the 70 kDa antigen, recognized by the standard positive serum sample. These results suggest that the WB can be effectively used for the routine serodiagnosis of viral caprine arthritis encephalitis infection (CAEV). It can also support further studies on antigen recognition patterns with the new antigen provided by the novel simplified purification system of viral components described here.

RESUMO O sorodiagnóstico das lentiviroses de caprinos e ovinos é realizado principalmente pela imunodifusão em gel de agar (AGID) e/ou ELISA. Embora relativamente simples, esses métodos não identificam os antígenos virais reconhecidos na resposta imune do animal examinado, por isso o western blot (WB) vem ganhando maior relevância como ferramenta de diagnóstico dessas enfermidades. Neste trabalho, o antígeno utilizado no WB foi obtido através de um sistema simplificado de purificação: concentração por diálise do sobrenadante de culturas celulares infectadas, seguido de centrifugação em gradiente contínuo de sacarose. A separação das proteínas virais foi obtida por SDS-PAGE a 10% e a transferência para membranas de nitrocelulose realizada pelo sistema semi-úmido. A revelação das membranas mostrou reconhecimento pelo soro padrão positivo de cinco proteínas, com pesos moleculares de 14-16, 25, 40, 50 e 70 kDa. Todas as 8 amostras de soro caprino, positivas na ADIG, reconheceram pelo menos uma banda proteíca no immunoblot, variando contudo o número de bandas reconhecidas. Reação positiva à glícoproteína 40 (gp 40) foi observada em quatro animais, com intensidade de reação discreta em três deles. Dois animais apresentaram reação positiva à proteína 16 (p16), e dois à gp 50, de pouca intensidade. Quanto à gp 70, proteína que, embora reagisse com o soro padrão positivo, não foi reconhecida por nenhum dos soros testados. Estes resultados sugerem que o WB pode ser empregado para o sorodiagnóstico rotineiro das lentiviroses, ensejando estudos mais amplos do padrão de reconhecimento dos antígenos apresentados por este novo sistema de purificação parcial de componentes virais.

Lentivírus de pequenos ruminantes em ovinos santa inês: isolamento, identificação pela pcr e inquérito sorológico no estado de pernambuco

ABSTRACT Small ruminants can be infected by a group generically denominated as small ruminant lentiviruses (SRLV), which comprehend several isolates, distributed into four phylogenetic groups. The Maedi-Visna (MVV) and caprine arthritis encephalitis (CAEV) viruses, originally isolated from ovine and caprine, respectively, are the prototypes of both groups. The objective of this work was to isolate and identify an SRLV strain and carry out a serological survey in ovines of the Santa Inês breed. Through co-cultivation of ovine sinovial cells and leucocytes of a seropositive ewe, LVSR was isolated (BrPe2-01 strain), and identified by the appearance of syncitium on the monolayers and by polymerase chain reaction (PCR). The serology was done through the agar gel immunodiffusion test (AGID) in 558 sheep of 25 flocks from the state of Pernambuco, Brazil. The results demonstrated positive serology in 6 (1.07%) sheep of 3 (12%) studied flocks. It was concluded that the ovine of the Santa Inês breed are being infected by SRLV, with low prevalence.

RESUMO Os pequenos ruminantes podem ser infectados por um grupo de vírus genericamente denominado de Lentivírus de Pequenos Ruminantes (LVPR), que compreende vários isolados, distribuídos em quatro grupos filogenéticos. Os vírus Maedi-Visna (MVV) e da Artrite Encefalite Caprina (CAEV), originalmente, isolados de ovinos e caprinos, respectivamente, são os protótipos dos dois grupos. O objetivo desse trabalho foi isolar, identificar amostra de LVPR e realizar levantamento sorológico, em ovinos da raça Santa Inês. Através do co-cultivo de células de membrana sinovial ovina e sangue periférico de um ovino soropositivo ao teste de IDGA foi isolado LVPR, amostra BrPe2-01, identificada pelo aparecimento de sincícios nas monocamadas e pela reação em cadeia da polimerase (PCR). A sorologia foi feita através do teste de Imunodifusão em Gel de Agar (IDGA) em animais de 25 propriedades procedentes das regiões Agreste, Sertão, Mata Meridional e Metropolitana do Estado de Pernambuco, totalizando 558 amostras. Os resultados demonstraram sorologia positiva em 6 (1,07%) ovinos de 3 (12%) rebanhos estudados. Concluiu-se que os ovinos da raça Santa Inês estão sendo infectados pelos LVPR, porém com baixa prevalência.

Anticorpos contra lentivírus de pequenos ruminantes em caprinos e ovinos em abatedouros do estado de Pernambuco / Small ruminant lentivirus infection in goats and sheep from two abattoirs in Pernambuco State, Brazil

The prevalence of small ruminant lentivirus (SRL) infection was evaluated in goats and sheep in two counties of Pernambuco State, Brasil. Seriological examinations were performed from a total of 672 goats and 325 sheep, one year of age and older, at two abattoirs. Sera were analyzed by agar gel immunodiffusion using Maedi-Visna K-1514 antigens. There were 42 reactive samples (95 percent confidence interval 3.6 percent to 4.9 percent) in both slaughter houses. In São Lourenço da Mata county, 3.2 percent and 4.0 percent of goat and sheep sera were responsive, whereas in Paulista county, 5.1 percent of goat samples and 8.2 percent of sheep were reactive. Thus, the prevalence of small ruminant lentivirus was low in goats and sheep sampled from the region.

Anticorpos contra lentivírus de pequenos ruminantes em caprinos e ovinos em abatedouros do estado de Pernambuco / Small ruminant lentivirus infection in goats and sheep from two abattoirs in Pernambuco State, Brazil

The prevalence of small ruminant lentivirus (SRL) infection was evaluated in goats and sheep in two counties of Pernambuco State, Brasil. Seriological examinations were performed from a total of 672 goats and 325 sheep, one year of age and older, at two abattoirs. Sera were analyzed by agar gel immunodiffusion using Maedi-Visna K-1514 antigens. There were 42 reactive samples (95% confidence interval 3.6% to 4.9%) in both slaughter houses. In São Lourenço da Mata county, 3.2% and 4.0% of goat and sheep sera were responsive, whereas in Paulista county, 5.1% of goat samples and 8.2% of sheep were reactive. Thus, the prevalence of small ruminant lentivirus was low in goats and sheep sampled from the region.(AU)

In vitro cytochemical studies of the golgi apparatus of pancreatic acinar cells exposed to drugs that inhibit the transport of membranes and proteins.

In a preceding article, we described alterations occurring in rat pancreas acinar cells at successive post-mortem (PM) intervals. In ultra-thin sections from samples obtained from 0.5, 1, 2, 4, 8 and 12 h, we observed in the Golgi apparatus the appearance of an anomalous membrane bound structure. Such structures are formed by tubules and vesicles that we have called tubular vesicular structure (TVS), and they are frequently located in the position corresponding to the 4th cisterna of the Golgian cisternal pile. Lobules of rat pancreas, incubated in vitro with metabolic inhibitors (such as antimycin A, sodium fluoride, sodium azide and potassium cyanide), were processed in order to be compared with the PM samples of the rat acinar cells. In sliced pieces of lobules, acid phosphatase (AcPase) and tiaminopirophosphatase (TPPase) activity were evaluated. Except for the potassium cyanide treatment, we frequently observed the TVS located at the position corresponding to the 4th cisternae (similar to those observed in the PM acinar cells). These TVS's are predominantly TPPase positive. Based on this result and the fact that the TVS's are surrounded by a membrane (as confirmed by the freeze-fracture replica results) with no structural elements inside, they seem not to correspond to autophagosomes. The TVS's, observed either at PM consecutive times or incubated with metabolic inhibitors, seem to be structures formed in response to ATP deprivation. In 0,5 h PM cells and in cells incubated for 30 and 60 min with metabolic inhibitors, the subcellular structures reacted for AcPase in the rigid lamellae, CV and lysosomes.

Evidência sorológica de infecçäo pelo vírus da artrite-encefalite caprina em caprinos leiteiros do Estado de Pernambuco / Sorological evidence of caprine arthritis-encephalitis in dairy goats in Brazil

Foram pesquisados anticorpos precipitantes contra o vírus da artrite-encefalite caprina (CAE), empregando-se a imunodifusäo em ágar-gel, utilizando-se o antígeno do vírus maedi/visna dos ovinos, em 214 amostras de soros de caprinos puros (Saaneu, Toggenburg e Parda Alpina) e seus mestiços, provenientes de cinco exploraçöes leiteiras de quatro municípios do Estado. Evidenciaram-se resultados positivos em 38 amostras (17,7 por cento), com frequências por criaçäo variando de zero a 29,5 por cento

Anticorpos contra pestivirus e herpesvirus em caprinos leiteiros no Estado de Pernambuco / Antibodies against pestivirus and herpesvirus in dairy goats in Pernambuco State, Brazil

A preliminar survey to detect antibodies against BVD/MD (NADL) and BHV1 (Los Angeles) was perfomed in five dairy goat flocks from four counties of Pernambuco State, Brazil. Two hundred and six serum samples of purebred (Saanen, Toggenburg and Alpine) and crossbred goats were tested by microplate serumneutralization test. Positive results (titre >/ 4) were recorded in 11.6 per cent and 6.8 per cent of samples test to BVD/MD and BHV 1, respectively

[The use of sodium azide as a fecal preservative for studying the eggs of Schistosoma mansoni and of other helminths by the Kato-Katz method: a study in an endemic area]. / Emprego da azida sódica, como conservador de fezes, para a pesquisa de ovos de Schistosoma mansoni e de outros helmintos, pelo método de Kato-Katz: estudo em área endêmica.

In two aliquots of 208 samples of stool, tests have been made by the Kato-Katz method, one being made after the use of 0.2mg of sodic azide for 200mg, and the other without the aforesaid conservant. The resulted percentages with or without the conservant were, respectively, for Ancylostomideos: 12.5 and 25.9; for Ascaris lumbricoides: 71.6 and 72.5; for Schistosoma mansoni: 7.6 and 17.7, and for Trichuris trichiura: 86 and 85. The count of the eggs with and without the conservant was, respectively, 264 and 539 for Ancylostomideos, 13186 and 33751 for A. lumbricoides, 55.5 and 63.5 for S. mansoni, and 1345 and 2068 for T. trichiura. The authors did not confirm the advantage of using sodic azide for study in endemic areas. They suggest that the exsiccation of the stool and the low intensity of infections can explain the unfavourable results of the present clinical trial.

[The possible therapeutic effect of deferoxamine in experimental infections of mice by Plasmodium berghei]. / Estudo sobre a possível atividade terapêutica da deferoxamina na infecção experimental de camundongos pelo Plasmodium berghei.

Deferoxamine is a compound with iron chelating properties. Body depletion of this mineral, according to some authors, might influence the metabolism of plasmodia that thrive in the erythrocytes. In order to verify this possibility, we administered daily doses of 300 or 1.000 mg/kg of the compound, for five days and again 15 days, to mice infected with Plasmodium berghei. The parameters used to check the activity of deferoxamine were mortality of the animals and the count of blood parasites. The results showed a progressive increase of mortality and of the parasitemia in all animals, without differences in relation to the controls. So, at least on hand of the present study, the mentioned substance cannot be considered useful for the treatment of malaria.

[Viability of Trypanosoma cruzi in sugar cane juice]. / Observações sobre a viabilidade do Trypanosoma cruzi no caldo da cana.

In Catolé do Rocha, State of Paraíba, Brasil several people were concomitantly contaminated by Chagas' disease and had the acute form of the infection. Because of this event, the hypothesis that Trypanosoma cruzi might have been transmitted by the digestive route, through the ingestion of accidentally contaminated sugar cane juice, was made. In the present study, we verified that the parasite has remained viable for four hours in the liquid, at room temperature. We have also demonstrated that, after one hour, it was capable of promoting infection in one mouse, by gastric tube administration. These observations enhance the interest on this alternative mechanism of disease transmission.

[Evaluation of the therapeutic effect of albendazole on experimental and human infections by Hymenolepis nana]. / Avaliação da atividade terapêutica do albendazol sobre infecções experimentale e humana pela Hymenolepis nana.

With the objective of knowing adequately the spectrum of activity of albendazole against intestinal helminthiases, we made observations regarding hymenolepiasis caused by Hymenolepis nana. Two series of investigations were carried out: a) treatment of mice with single doses of either 25 or 50 mg/kg, repeated after ten days, using as controls animals treated with 25 mg/kg of praziquantel or not treated with any antiparasitic drugs; b) treatment of adults and children with 400 mg daily for three consecutive days, repeated after ten days.

[Evaluation of the usefulness of the Coprotest for parasitologic examination of feces]. / Avaliação da utilidade do "Coprotest" para exame parasitológico das fezes.

"Coprotest" is a device used for parasitological examination of the stools that has been marketed in Brazil with no previous literature support. Therefore, we decided to evaluate the usefulness of the procedure using "Coprotest", comparing it with the routine methods by Hoffman et al., Faust et al., and Rugai et al. Taking into consideration only positive samples, we observed the following figures, according to each technique: Hofmann et al.--98 (47.34%); Faust et al.--119 (57.48%); Rugai et al., only for larva detection--21 (10.14%); "Coprotest"--115 (55.55%). We concluded that "Coprotest" in addition to its operational qualities, has the merits that allow its use together with other techniques for parasitosis diagnosis. Nevertheless it has not modified the previous situation, since more than one technique must still be used, as we have noted for detection of Strongyloides stercoralis larvae in this instance.