Dilmara Reischak

Phylogenetic analysis of small ruminant lentiviruses from Southern Brazil

The first lentivirus isolated from sheep in Brazil was analysed phylogenetically. Evolutionary trees of the proviral 597 nucleotide gag and 432 nucleotide pol sequences obtained by the maximum likelihood method demonstrated that the sheep isolate clustered with prototype Maedi Visna virus whereas three lentiviruses isolated from goats in the same geographic region were close to caprine arthritis encephalitis prototypes. A subsequent comparison of sequence data of these viruses with those contained in the EMBL sequence database revealed that, in contrast to caprine prototypic viruses, all prototypic Maedi Visna viruses contain a deletion of six nucleotides in the gag gene resulting in the deletion of two residues in the central region of capsid protein. This deletion may be a useful marker in the analysis of small ruminant lentiviruses, especially when considering possible transmission of lentiviruses between sheep and goats.

Detecção da infecção pelo vírus da artrite-encefalite caprina: imunodifusão em ágar e reação em cadeia da polimerase com "primers" degenerados

The purpose of this work was to analyse serum and blood cells from caprine origin to detect antibody and proviral DNA of caprine arthritis encephalitis virus (CAEV), respectively. Agar gel immunodiffusion (AGID) and polymerase chain reaction (PCR) with degenerated primers were used. Samples of different geographical regions were analysed: 39 from Mato Grosso do Sul (MS), 19 from Sao Paulo (SP), 22 from Ceara (CE) including 10 from Canada (imported animals), providing a total of 80 samples. The results obtained by AGID and PCR were discordants, as 25 samples were detected as seropositive, while 16 infected animals were detected by PCR. On the other hand, PCR allowed the identification of infected animals that did not have detectable antibodies by AGID: eight samples from MS and one from CE. Different aspects related to these discordant results are discussed.

Imunofluorescência utilizando isolados brasileiros no diagnóstico sorológico de infecção por lentivírus em caprinos

Small ruminant lentiviruses (SRLV) are distributed worldwide and cause persistent infections in sheep and goats. The purpose of this work was to develop an indirect immunofluorescent antibody (IFA) test using Brazilian SRLV isolates for serological diagnosis of infection in goats. The IFA test was compared in its sensitivity and specificity to the AGID test in which Maedi-Visna WLC-1 strain was used as antigen. Ovine synovial secondary cell cultures infected with two goat SRLV isolates (CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5) were used for the IFA. Goat serum samples (n = 239) were tested by the two tests. The AGID test detected antibodies in 129 (53.9%) serum samples. A higher number of animals was considered positive for the IFA. However, different results were obtained depending on the isolate used for the antigen preparation. When CAEV UFRGS/Br-2 was used as antigen, 216 (90.3%) serum samples tested positive, against 213 (89.1%) with CAEV UFRGS/Br-5. There was no significant statistical difference between the antigens prepared with these two isolates. The IFA had sensitivity of 94.6% and 96.9%, and specificity of 14.5 % and 20%, when CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5 were used as antigens, respectively. These results demonstrate that the IFA with antigens prepared with Brazilian SRLV isolates may play an important a role as a complementary serological test for the diagnosis of infections by these agents.