Use of a commercial ELISA kit specific for glycoprotein E peptides to indirectly detect Caprine Herpesvirus 1 (CpHV-1) in the state of São Paulo, Brazil / Uso de kit ELISA comercial específico para peptídeos da glicoproteína E para detecção indireta de Herpesvírus Caprino tipo 1 (CpHv-1) no estado de São Paulo, Brasil

Caprine herpesvirus 1 (CpHV-1) infection is associated with clinical manifestations related to animal age, with high mortality in kids and infertility in adults. Given the scarcity of research about the epidemiological situation of this infection in Brazilian flocks, we aimed to conduct a cross-sectional descriptive study to detect antibodies against CpHV-1 in goats in the state of São Paulo, Brazil. Fifty-five male and female goats ­ kids and adult ­ were assessed in this study. Blood serum was analyzed by a commercial ELISA kit to detect antibodies against CpHV-1, which had not been used in Brazil before. No animals were reactive. Brazil lacks information about CpHV-1 infection in goat flocks. Continuing the study is crucial to understand the epidemiological situation of the disease and establish protocols for infection control.(AU)

A infecção pelo Herpesvírus Caprino tipo 1 (CpHv-1) está associada a manifestações clínicas relacionadas à idade dos animais, com alta mortalidade em filhotes e infertilidade em adultos. Diante da escassez de estudos sobre situação epidemiológica dessa infecção nos rebanhos brasileiros, a presente pesquisa teve como objetivo realizar um estudo transversal e descritivo para a detecção de anticorpos anti-Herpesvírus Caprino tipo 1 em caprinos do estado de São Paulo, Brasil. Foram avaliados 55 caprinos machos e fêmeas, filhotes e adultos. O soro sanguíneo foi analisado por um kit ELISA comercial para detecção de anticorpos contra CpHv-1, de utilização inédita no Brasil. Nenhum animal estudado foi sororreagente. O Brasil carece de informações acerca da infecção pelo Herpesvírus Caprino tipo 1 nos rebanhos caprinos do país. A continuidade do estudo é imprescindível para compreender a situação epidemiológica da enfermidade e estabelecer protocolos para controle da infecção.(AU)

Use of a commercial ELISA kit specific for glycoprotein E peptides to indirectly detect Caprine Herpesvirus 1 (CpHV-1) in the state of São Paulo, Brazil / Uso de kit ELISA comercial específico para peptídeos da glicoproteína E para detecção indireta de Herpesvírus Caprino tipo 1 (CpHv-1) no estado de São Paulo, Brasil

Caprine herpesvirus 1 (CpHV-1) infection is associated with clinical manifestations related to animal age, with high mortality in kids and infertility in adults. Given the scarcity of research about the epidemiological situation of this infection in Brazilian flocks, we aimed to conduct a cross-sectional descriptive study to detect antibodies against CpHV-1 in goats in the state of São Paulo, Brazil. Fifty-five male and female goats ­ kids and adult ­ were assessed in this study. Blood serum was analyzed by a commercial ELISA kit to detect antibodies against CpHV-1, which had not been used in Brazil before. No animals were reactive. Brazil lacks information about CpHV-1 infection in goat flocks. Continuing the study is crucial to understand the epidemiological situation of the disease and establish protocols for infection control.(AU)

A infecção pelo Herpesvírus Caprino tipo 1 (CpHv-1) está associada a manifestações clínicas relacionadas à idade dos animais, com alta mortalidade em filhotes e infertilidade em adultos. Diante da escassez de estudos sobre situação epidemiológica dessa infecção nos rebanhos brasileiros, a presente pesquisa teve como objetivo realizar um estudo transversal e descritivo para a detecção de anticorpos anti-Herpesvírus Caprino tipo 1 em caprinos do estado de São Paulo, Brasil. Foram avaliados 55 caprinos machos e fêmeas, filhotes e adultos. O soro sanguíneo foi analisado por um kit ELISA comercial para detecção de anticorpos contra CpHv-1, de utilização inédita no Brasil. Nenhum animal estudado foi sororreagente. O Brasil carece de informações acerca da infecção pelo Herpesvírus Caprino tipo 1 nos rebanhos caprinos do país. A continuidade do estudo é imprescindível para compreender a situação epidemiológica da enfermidade e estabelecer protocolos para controle da infecção.(AU)

Molecular detection of viral agents in free-ranging and captive neotropical felids in Brazil.

We describe molecular testing for felid alphaherpesvirus 1 (FHV-1), carnivore protoparvovirus 1 (CPPV-1), feline calicivirus (FCV), alphacoronavirus 1 (feline coronavirus [FCoV]), feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and canine distemper virus (CDV) in whole blood samples of 109 free-ranging and 68 captive neotropical felids from Brazil. Samples from 2 jaguars ( Panthera onca) and 1 oncilla ( Leopardus tigrinus) were positive for FHV-1; 2 jaguars, 1 puma ( Puma concolor), and 1 jaguarundi ( Herpairulus yagouaroundi) tested positive for CPPV-1; and 1 puma was positive for FIV. Based on comparison of 103 nucleotides of the UL24-UL25 gene, the FHV-1 sequences were 99-100% similar to the FHV-1 strain of domestic cats. Nucleotide sequences of CPPV-1 were closely related to sequences detected in other wild carnivores, comparing 294 nucleotides of the VP1 gene. The FIV nucleotide sequence detected in the free-ranging puma, based on comparison of 444 nucleotides of the pol gene, grouped with other lentiviruses described in pumas, and had 82.4% identity with a free-ranging puma from Yellowstone Park and 79.5% with a captive puma from Brazil. Our data document the circulation of FHV-1, CPPV-1, and FIV in neotropical felids in Brazil.

Genome sequence of bubaline alphaherpesvirus 1 (BuHV1) isolated in Australia in 1972.

Bubaline alphaherpesvirus 1 (BuHV1) is a member of the family Herpesviridae, subfamily Alphaherpesvirinae, genus Varicellovirus. To date, no full genome sequence of BuHV has been published. Here, we report the complete genome sequence of bubaline alphaherpesvirus 1 (BuHV1) strain b6 (BuHV1-b6), isolated from a water buffalo (Bubalus bubalis) in 1972 in Australia. The virus was multiplied in MDBK cells, and the DNA was extracted and subjected to high-throughput sequencing. The reads were aligned and combined into a single genome sequence, with bovine alphaherpesvirus 5 (BoHV5) strain SV507/99 (accession number NC005261) as a reference. The BuHV1-b6 genome is a linear double-stranded DNA molecule, 137,452 bp long, with a GC content of 76.8%. The genome consists of two unique sequences: a long, or UL, sequence (103,818 bp) and a short, or US, sequence (9,586 bp), with the latter being flanked by inverted IR and TR elements of 12,024 bp each. The arrangement is typical of herpesvirus genomes of the D-type. The overall sequence has a 92.2% similarity at the nucleotide level to the reference BoHV5 strain. Our report provides a significant landmark in the history of herpesviruses, represented by the genome sequence of this 44-year-old virus isolate.

Bovine herpesvirus 6 in buffaloes (Bubalus bulalis) from the Amazon region, Brazil.

This study presents the first description of Bovine herpesvirus 6 (BoHV-6) that was isolated from buffaloes of Amazon region in Brazil. Phylogenetic analysis showed that the BoHV-6 Brazilian strains clustered with the sequence of BoHV-6 from elsewhere available at the GenBank. It was observed in some buffaloes with lymphoproliferative disease in one herd, thus the animals were also tested for Bovine leukemia virus (BLV), which has been associated to lymphoma in bovines. All animals were negative to BLV. These results indicate that BoHV-6 is present in buffaloes in Brazil, but the importance and impact of this infection and its association with any illness is still undefined.

Evaluation of prenucleic acid extraction for increasing sensitivity of detection of virus in bovine follicular fluid pools.

Bovine viral diarrhea virus (BVDV), bovine herpesvirus type 1 (BoHV-1), and bovine herpesvirus type 5 (BoHV-5) are major cattle pathogens that can be present in biological materials used in assisted reproduction biotechnologies. The aim of the present study was to increase the sensitivity of the polymerase chain reaction (PCR) for detection of BVDV, BoHV-1, and BoHV-5 in bovine follicular fluid (FF) collected during oocyte retrieval for in vitro embryo production. Ovaries were collected immediately after slaughter at a commercial abattoir, aspirated, and the 7336 samples of FF were pooled in 84 samples. Before testing the FF field samples, sensitivity of the protocol was determined using a prenucleic acid extraction procedure that was directly compared with standard RNA or DNA extraction protocols. The prenucleic acid extraction procedure increased sensitivity of reverse transcription (RT)-PCR for BVDV and nested PCR for BoHV-1 and BoHV-5 by 100 and 10 times, respectively. The 84 FF pools were assayed for BVDV, BoHV-1, and BoHV-5 using virus isolation and RT-PCR or nested PCR. Fourteen (16.7%) FF pools were positive for BVDV RNA, and one (1.2%) was positive for BoHV-1 DNA. Two of the BVDV RT-PCR positive samples and the one BoHV-1 PCR positive sample were also positive in cell culture, demonstrating that FF contained infectious viruses. In this study, the prenucleic acid extraction procedure increased the sensitivity of RT-PCR and PCR detection. This study highlighted the importance of assuring biosecurity by detecting the presence of viral pathogens in biological materials used during in vitro embryo production.

Serological evidence and risk factors associated with Caprine herpesvirus 1 in dairy goat flocks in a semiarid region of northeastern Brazil.

A cross-sectional study was carried out to determine the flock-level seroprevalence of Caprine herpesvirus 1 (CpHV-1) and Bovine herpesvirus 1 (BoHV-1) and 2 (BoHV-2) and risk factors associated with CpHV-1 in dairy goat flocks from a semiarid region of northeastern Brazil. A total of 1,034 serum samples from 110 flocks were collected from March 2009 through March 2010. A structured questionnaire focusing on variables related to risk factors for CpHV-1 infection was given to each farmer at the time of blood collection. Antibodies against CpHV-1, BoHV-1, and BoHV-2 were detected by neutralization tests. The flock-level prevalences of CpHV-1, BoHV-1, and BoHV-2 were 89.1% (98/110; 95% confidence interval [CI]: 81.7-94.2), 80% (88/110; 95% CI: 71.3-87), and 4.5% (5/110; 95% CI: 1.5-10.3), respectively. Frequencies of seropositive animals were 36.6% (379/1,034), 25.8% (267/1,034), and 0.6% (6/1,034) for CpHV-1, BoHV-1, and BoHV-2, respectively. The use of natural mating was identified as a risk factor associated with CpHV-1 flock-level prevalence (P = 0.001). It is suggested that adoption of veterinary services and active surveillance of the at-risk flocks in the study region should be initiated to reduce the prevalence of herpesvirus infections.

Development of a differential multiplex PCR assay for equine herpesvirus 1 and 4 as a diagnostic tool.

In this study, a multiplex polymerase chain reaction (PCR) procedure was developed for differentiation of strains and field isolates of equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4). Specific oli-gonucleotide primers were combined to amplify the thymidine kinase (TK) gene region of EHV-1 and EHV-4, which would yield fragments of different lengths for each virus in the same amplification reaction. The specificity of the largest PCR amplicon for EHV-4 was confirmed by restriction digestion with HindIII. The multiplex PCR proved to be a fast and sensitive method for typing EHV-1 and EHV-4 isolates and for detection and differentiation of both viruses in field samples in which infectious virus is no longer available. The sensitivity was improved by combining cycling optimization and visualization of PCR products in ethidium bromide and silver-stained acrylamide gels.