Genotypic characterization of psittacid herpesvirus isolates from Brazil

Abstract Thirty-six isolates of psittacid herpesvirus (PsHV), obtained from 12 different species of psittacids in Brazil, were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis and PCR amplification. RFLP analysis with the PstI enzyme revealed four distinct restriction patterns (A1, X, W and Y), of which only A1 (corresponding to PsHV-1) had previously been described. To study PCR amplification patterns, six pairs of primers were used. Using this method, six variants were identified, of which, variants 10, 8, and 9 (in this order) were most prevalent, followed by variants 1, 4, and 5. It was not possible to correlate the PCR and RFLP patterns. Twenty-nine of the 36 isolates were shown to contain a 419 bp fragment of the UL16 gene, displaying high similarity to the PsHV-1 sequences available in GenBank. Comparison of the results with the literature data suggests that the 36 Brazilian isolates from this study belong to genotype 1 and serotype 1.

Genotypic characterization of psittacid herpesvirus isolates from Brazil.

Thirty-six isolates of psittacid herpesvirus (PsHV), obtained from 12 different species of psittacids in Brazil, were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis and PCR amplification. RFLP analysis with the PstI enzyme revealed four distinct restriction patterns (A1, X, W and Y), of which only A1 (corresponding to PsHV-1) had previously been described. To study PCR amplification patterns, six pairs of primers were used. Using this method, six variants were identified, of which, variants 10, 8, and 9 (in this order) were most prevalent, followed by variants 1, 4, and 5. It was not possible to correlate the PCR and RFLP patterns. Twenty-nine of the 36 isolates were shown to contain a 419bp fragment of the UL16 gene, displaying high similarity to the PsHV-1 sequences available in GenBank. Comparison of the results with the literature data suggests that the 36 Brazilian isolates from this study belong to genotype 1 and serotype 1.

Outbreak of human herpesvirus type 1 infection in nonhuman primates (Callithrix penincillata).

Human herpesvirus type 1 (HHV-1) is widely dispersed among the human population. Although infection is often asymptomatic in humans, nonhuman primates develop a severe and often fatal infection. In August 2006, 13 black-tufted marmosets (Callithrix penincillata) from a group of 14 presented with clinical apathy, anorexia, and ataxia. Physical examination revealed conjunctivitis, erosive or ulcerative lesions on the skin, and swollen lymph nodes. Of the 14 animals captured, 10 died. Grossly, ulcers and erosions were observed on the skin of face, nasal planum, lips, and oral mucosa. Histologically, superficial vesicular and erosive stomatitis with associated basophilic intranuclear inclusion bodies in the squamous epithelium were observed. Swabs from oral lesions and tissue samples from necropsied animals were positive for HHV-1 by nested polymerase chain reaction for eight animals.

Identification and isolation of psittacid herpesvirus from psittacids in Brazil.

Psittacid herpesvirus (PsHV) was isolated from 41 birds kept in captivity in Belo Horizonte, Minas Gerais/Brazil using chicken embryo fibroblasts (CEF) cell cultures. For this study, leukocytes or cloacal swabs of live birds were used. Also, portions of liver, spleen or kidney from birds collected at necropsy were utilized for these tests. PCR tests confirmed the presence of PsHV in 100% of samples. Thirty-three of the PCR products were sequenced and the results disclosed a 99% and 100% identity when compared with other sequences PsHV-1 (AY372243.1 and AF261756.1), previously deposited in GenBank. In addition, histopathology was performed and 19 of the 29 birds contained random multifocal lymphoplasmacytic hepatitis with necrotic foci, suggestive of viral infection. Three samples were examined by electron microscopy to visualize the viral particles obtained from cell culture. The viral structures measured 269 nm in average, had envelopes with an icosahedral capsid and tegument, consistent with herpesvirus. Thus, a total of 41 isolates were obtained from PsHV cell cultivation in CEF, confirming the circulation of the virus between parrots kept in captivity in Belo Horizonte, and affirming the importance of further studies in this area.

Naturally occurring feline leukemia virus subgroup A and B infections in urban domestic cats.

A nested-PCR (n-PCR) was used to detect feline leukemia virus (FeLV) proviral DNA in blood samples from 464 sick and 608 healthy domestic cats (Felis catus) selected by convenience, and a significantly high prevalence of FeLV infection was observed. n-PCR results revealed the presence of FeLV proviral DNA in 47.2 % of sick cats and 47.4 % of healthy cats. Phylogenetic analysis revealed that FeLV samples from healthy or sick cats were grouped into separate clades. We determined FeLV subgroups by an n-PCR based on the envelope (env) gene. The partial env gene of FeLV Minas Gerais (MG) samples were compared to various exogenous FeLV isolates and endogenous (enFeLV) provirus from the same region. FeLV-B MG samples were more similar to endogenous sequences and to natural FeLV-B isolates than to either FeLV-A or FeLV-C. The results revealed the circulation of FeLV-B in large populations of urban domestic cats in Brazil.