Raymundo Rizaldo Pinheiro

Risk factors for infection by Toxoplasma gondii in herds of goats in Ceará, Brazil

In order to identify possible risk factors for T. gondii infection in goat herds in Ceara, Brazil, 2362 serum samples were tested by ELISA. The serological prevalence was 25.1%. The risk factors identified for Toxoplasma gondii infection in goat herds were age, number of cats, use of wooden feeding troughs and absence of feeding troughs. Goats older than 37 months had 2.01 (CI 95%; 1.55 - 2.61) higher risk of infection than younger animals. Greater risk of infection was observed in farms with more than 10 cats (OR = 1.73; CI 95%; 1.01 - 3.33). The use of wooden feeding troughs represented a high probability of infection (OR = 7.81; CI 95%; 1.66 - 36.67). The lack of feeding troughs also represented a high probability of infection (OR = 5.50; CI 95%; 1.24 - 24.39).

Fatores de risco na transmissão do lentivírus caprino pelo sêmen

The objective of this work was to evaluate the presence of the DNA provirus of the caprine lentivirus (LVC) in ejaculates of naturally infected males, and to verify the influence of the wash of the semen as well as the presence of testicle inflammation on the viral load. Eight semen collections of seven soropositive reproducers were accomplished, four before testicle injury and four after injury. Amongst the collections carried out at the same week, in one the ejaculate was washed, to withdraw the plasma seminal, and in the other it was not. The provirus DNA was identified both by Nested polymerase chain reaction technique (Nested PCR) and by the viral isolation. The virus was isolated in 7.1% of the samples. The PCR identified the provirus DNA in 35.7% of all samples, 17.9% in the washed samples and 53.6% of the integral semen samples. The injury of the testicle tends to greater flow of virus for the semen, therefore, before injury, 21.4% of the samples were positive and after-injury, 50%. Risk of transmission of the LVC by semen of goat reproducers is strengthened by the presence of testicle inflammations and the fact that the criopreserved semen contains the LVC in infecting form.

Associação entre o California Mastitis Test e a Contagem de Células Somáticas na avaliação da saúde da glândula mamária caprina

A correlacao entre os resultados do California Mastitis Test (CMT) e da Contagem de Celulas Somaticas (CCS) foi estudada em 68 cabras das racas Anglo-Nubiana, Pardo Alpina e Saanen. Os dados foram coletados quinzenalmente, durante sete meses, realizando-se a CCS em amostras de leite bacteriologicamente negativas. Os resultados mostraram uma correlacao positiva e significativa (p < 0,05) entre os testes avaliados, com um coeficiente de r = 0,63. Quando os testes foram correlacionados com a producao de leite, observou-se uma correlacao negativa e significativa (p < 0,05) de r = -0,27 e r = -0,28 para o CMT e a CCS, respectivamente. Na associacao das reacoes do CMT com a CCS obtiveram-se medias de 0,78 x 106 cels/ml para as reacoes negativas (N, T e 1+) e media de 5,32 x 106 cels/ml para as reacoes positivas (2+ e 3+). Os resultados indicam que, para evitar resultados falso-positivos, outros testes diagnosticos deverao ser usados juntamente com o CMT, para se avaliar a saude da glândula mamaria caprina.

Interspecific transmission of small ruminant lentiviruses from goats to sheep

This study was conducted in order to evaluate the transmission of caprine lentivirus to sheep using different experimental groups. The first one (colostrum group) was formed by nine lambs receiving colostrum from goats positive for small ruminant lentiviruses (SRLV). The second group (milk group) was established by nine lambs that received milk of these goats. Third was a control group, consisting of lambs that suckled colostrum and milk of negative mothers. Another experimental group (contact group) was formed by eight adult sheep, confined with two naturally infected goats. The groups were monitored by immunoblotting (IB), enzyme-linked immunosorbent assay (ELISA), agar gel immunodiffusion (AGID) and nested polymerase chain reaction (nPCR). All lambs that suckled colostrum and milk of infected goats and six sheep of the contact group had positive results in the nPCR, although seroconversion was detected only in three of the exposed animals, with no clinical lentiviruses manifestation, in 720 days of observation. There was a close relationship between viral sequences obtained from infected animals and the prototype CAEV-Cork. Thus, it was concluded that SRLV can be transmitted from goats to sheep, however, the degree of adaptation of the virus strain to the host species probably interferes with the infection persistence and seroconversion rate.

Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina

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.

Detecção do vírus da Artrite Encefalite Caprina por nested PCR e nested RT-PCR em ovócitos e fluido uterino

Caprine arthritis-encephalitis (CAE) is an infectious disease caused by the caprine arthritis-encephalitis virus (CAEV), belonging to the lentivirus genus. The presence of the virus has been observed in the nervous system, respiratory tract and mammary gland, and also in the male and female genital tract. The objective of this study is to identify the virus in oocyte and uterine fluid of infected goats by molecular diagnostic techniques, in order to assess the possibility of CAEV transmission with reproduction. Thirteen infected goats were selected and submitted to euthanasia for the collection of the reproductive system, aspiration of the uterine fluid and dissection of ovaries for oocyte collection. In order to identify the CAEV in the collected material, in the protovirus and free forms, it was submitted to the nRT-PCR and nPCR techniques, respectively. As a result, it was observed that 53.8% of oocytes were positive to nRT-PCR, while only 9.1% were positive to nPCR. The nRT-PCR also identified the virus in the uterine fluid of 46.1% of the tested females. Even though the 13 goats had CAEV, 30.8% presented negative results in nPCR and nRT-PCR in all of the analyzed samples (oocyte and uterine fluid). This work concludes that nRT-PCR and nPCR can be used in the diagnosis of CAE for the analysis with oocytes and uterine fluid, and that the presence of CAEV in these materials points out to the risk of CAEV transmission through reproductive technologies used in females.

Inquérito soroepidemiológico do lentivírus caprino e perfil das criações de caprinos na região do Baixo Médio São Francisco (BA)

This work aimed to determine the serological prevalence of caprine artrithis encephalitis (CAE) in the microregion of Juazeiro, in the state of Bahia, using the technique of agar gel immunodiffusion (AGID) to characterize the farming systems of the region. We collected 693 blood serum samples of goats from 46 farms in 8 different locations belonging to the municipalities forming the microregion (Campo Alegre de Lourdes, Casa Nova, Curaca, Juazeiro, Pilao Arcado, Remanso, and Sobradinho). During the visits, a questionnaire was applied emphasizing information related to health management. All of the visited properties had extensive breeding systems, with predominance of mixed breed animals, low productivity and low technology rate, aiming mainly to use the meat. The main diseases reported were caseous lymphadenitis, diarrhea, ectoparasites and keratoconjunctivitis. Concerning seroprevalence, 0.29% (2/693) of samples tested positive for AGID. The positive animals belonged to the same property in the municipality of Curaca, which showed 12.5% (1/8) of positive properties, in contrast to 2.17% (1/46) of total seroprevalence in the other visited properties. These results therefore suggest the need for new epidemiological surveys in the region, especially at a time when the importation of animals for genetic improvement is taking place.

Inquérito soro-epidemiológico de Brucella ovis em rebanhos ovinos no semiárido baiano

In order to analyze the occurrence of antibodies to Brucella ovis in sheep in a semi-arid region, this survey was conducted in the micro-region of Juazeiro, Bahia State, Brazil. The agar gel immunodifusion test (AGID) was used to examine 694 serum samples of 58 flocks. Antibodies to B. ovis were found in 5 (0.72%) of the investigated animals. It is believed that this low number of positive cases is related to the characteristics of the production. The predominant system is the extensive one, with presence of locally adapted breeds and crossbred animals, aiming at the production of meat and skin, with low productivity rates and technical development.

Ocorrência de anticorpos contra o vírus Maedi-Visna em ovinos Santa Inês, no Estado de Sergipe, Brasil

Small Ruminant Lentiviruses (SRLV) are infectious diseases of viral etiology caused by retroviruses, characterized for being a slowly progressive degenerative disease. The aim of this study was to determine the occcurrence of antibodies against lentivirus in the flock of Santa Ines sheep in Sergipe, Brazil. A minimum number of samples was analyzed by the Astudillo calculation, with an expected prevalence (p) of 10%, error rate (α) of 20% and confidence level of 95% (g = 1.96) obtaining the minimum number of samples (n) of 861. Finally, 941 serum samples were collected, derived from 54 properties and 19 municipalities distributed in three regions of the State, and the number of collected samples was proportional to the sheep flock of each municipality. The serum was placed at -20°C until the Agar Gel Immunodiffusion test (AGID), and results considered as positive or suspicious in AGID were assessed by the Western blotting technique. Out of sampled animals, 194 (20.62%) were male and 747 (79.38%) were female. From the stratification by age, 300 were young sheep (31.88%), aged between six months and one year old, 308 (32.73%) were young adults, aged between 1 and 3 years old, and 333 (35.39%) were estimated to be older than three years old. Results showed that antibodies against the Maedi-Visna virus in the sheep flock occcurred in the State of Sergipe at low frequency (0.11%). Thus, the occcurrence of Maedi-Visna infection in sheep in the State of Sergipe is reported, and emphasizes the participation of the transit of animals in the epidemiology of the disease.

Caprine lentivirus in sheep milk and semen

Abstract: With the objective of detecting the presence of caprine lentivirus (CLV) in ewe milk and in ram semen, ten matrixes and four reproducers experimentally infected with CLV were used. Samples of ewe milk were collected during the four months of lactation, five collections per animal, totaling 50 samples. Regarding the rams, eight semen collections were made per animal, during one year of experimentation, totaling 32 samples. The milk and semen samples were submitted to DNA extraction and the nested polymerase chain reaction test (nPCR) to detect CLV proviral DNA. Eight (16%) of the milk samples were positive in nPCR originating from two ewes. Only one (3.12%) semen sample was positive. The amplification products were sequenced, and were confirmed to be a CLV genomic sequence. Thus, the presence of CLV proviral DNA in sheep milk and semen was demonstrated, confirming the feasibility of infection between species, and alerting to the risk of spreading infections. [Lentivirus caprino em leite e semen de ovinos]. Resumo: Com o objetivo de detectar a presenca do lentivirus caprino (LVC) no leite de ovelhas e no semen de carneiros, utilizaram-se 10 matrizes e quatro reprodutores infectados experimentalmente com o LVC. Foram coletadas amostras de leite das ovelhas durante os quatro meses de lactacao, ocorrendo cinco coletas por animal, totalizando 50 amostras. Quanto aos carneiros, realizaram-se oito coletas de semen por animal, durante um ano de experimentacao, totalizando 32 amostras. As amostras de leite e de semen foram submetidas a extracao de DNA e a prova de reacao em cadeia da polimerase do tipo nested (nPCR) visando a deteccao de DNA proviral do LVC. Oito (16%) amostras de leite foram positivas na nPCR oriundas de duas ovelhas. Apenas uma (3,12%) amostra de semen apresentou positividade. Produtos da amplificacao foram sequenciados, confirmando-se tratar de sequencia genomica do LVC. Dessa forma, demonstrou-se a presenca do DNA proviral do LVC em leite e semen de ovinos, confirmando a viabilidade da infeccao entre especies e, assim, alertando sobre o risco de que a infeccao seja disseminada.