Proposed Update to the Taxonomy of Pestiviruses: Eight Additional Species within the Genus Pestivirus, Family Flaviviridae.

Pestiviruses are plus-stranded RNA viruses belonging to the family Flaviviridae. They comprise several important pathogens like classical swine fever virus and bovine viral diarrhea virus that induce economically important animal diseases. In 2017, the last update of pestivirus taxonomy resulted in demarcation of 11 species designated Pestivirus A through Pestivirus K. Since then, multiple new pestiviruses have been reported including pathogens associated with disease in pigs or small ruminants. In addition, pestivirus sequences have been found during metagenomics analysis of different non-ungulate hosts (bats, rodents, whale, and pangolin), but the consequences of this pestivirus diversity for animal health still need to be established. To provide a systematic classification of the newly discovered viruses, we analyzed the genetic relationship based on complete coding sequences (cds) and deduced polyprotein sequences and calculated pairwise distances that allow species demarcation. In addition, phylogenetic analysis was performed based on a highly conserved region within the non-structural protein NS5B. Taking into account the genetic relationships observed together with available information about antigenic properties, host origin, and characteristics of disease, we propose to expand the number of pestivirus species to 19 by adding eight additional species designated Pestivirus L through Pestivirus S.

Herpesvirus surveillance and discovery in zoo-housed ruminants.

Gammaherpesvirus infections are ubiquitous in captive and free-ranging ruminants and are associated with a variety of clinical diseases ranging from subclinical or mild inflammatory syndromes to fatal diseases such as malignant catarrhal fever. Gammaherpesvirus infections have been fully characterized in only a few ruminant species, and the overall diversity, host range, and biologic effects of most are not known. This study investigated the presence and host distribution of gammaherpesviruses in ruminant species at two facilities, the San Diego Zoo and San Diego Zoo Safari Park. We tested antemortem (blood, nasal or oropharyngeal swabs) or postmortem (internal organs) samples from 715 healthy or diseased ruminants representing 96 species and subspecies, using a consensus-based herpesvirus PCR for a segment of the DNA polymerase (DPOL) gene. Among the 715 animals tested, 161 (22.5%) were PCR and sequencing positive for herpesvirus, while only 11 (6.83%) of the PCR positive animals showed clinical signs of malignant catarrhal fever. Forty-four DPOL genotypes were identified of which only 10 have been reported in GenBank. The data describe viral diversity within species and individuals, identify host ranges of potential new viruses, and address the proclivity and consequences of interspecies transmission during management practices in zoological parks. The discovery of new viruses with wide host ranges and presence of co-infection within individual animals also suggest that the evolutionary processes influencing Gammaherpesvirus diversity are more complex than previously recognized.

Delayed by Design: Role of Suboptimal Signal Peptidase Processing of Viral Structural Protein Precursors in Flaviviridae Virus Assembly.

The Flaviviridae virus family is classified into four different genera, including flavivirus, hepacivirus, pegivirus, and pestivirus, which cause significant morbidity and mortality in humans and other mammals, including ruminants and pigs. These are enveloped, single-stranded RNA viruses sharing a similar genome organization and replication scheme with certain unique features that differentiate them. All viruses in this family express a single polyprotein that encodes structural and nonstructural proteins at the N- and C-terminal regions, respectively. In general, the host signal peptidase cleaves the structural protein junction sites, while virus-encoded proteases process the nonstructural polyprotein region. It is known that signal peptidase processing is a rapid, co-translational event. Interestingly, certain signal peptidase processing site(s) in different Flaviviridae viral structural protein precursors display suboptimal cleavage kinetics. This review focuses on the recent progress regarding the Flaviviridae virus genus-specific mechanisms to downregulate signal peptidase-mediated processing at particular viral polyprotein junction sites and the role of delayed processing at these sites in infectious virus particle assembly.

Ibex-Associated Malignant Catarrhal Fever in Duikers (Cephalophus Spp).

Eight duikers, representing 3 different species cohoused in a single zoological collection, died in a 10-month period. Black, red-flanked, and yellow-backed duikers were affected, appearing clinically with a combination of anorexia, diarrhea, ataxia, tremors, and/or stupor, followed by death within 72 hours of onset of clinical signs. Consistent gross findings were pulmonary ecchymoses (8/8), generalized lymphadenomegaly (6/8), ascites (5/8), and pleural effusion (4/8). Dense lymphocyte infiltrates and arteritis affected numerous tissues in most animals. Ibex-associated malignant catarrhal fever (MCF) viral DNA was detected in all cases by polymerase chain reaction and in situ hybridization. Identical ibex-MCF virus sequence was detected in spleen of a clinically healthy ibex (Capra ibex) housed in a separate enclosure 35 meters away from the duikers.

Infection with Foamy Virus in Wild Ruminants-Evidence for a New Virus Reservoir?

Foamy viruses (FVs) are widely distributed and infect many animal species including non-human primates, horses, cattle, and cats. Several reports also suggest that other species can be FV hosts. Since most of such studies involved livestock or companion animals, we aimed to test blood samples from wild ruminants for the presence of FV-specific antibodies and, subsequently, genetic material. Out of 269 serum samples tested by ELISA with the bovine foamy virus (BFV) Gag and Bet antigens, 23 sera showed increased reactivity to at least one of them. High reactive sera represented 30% of bison samples and 7.5% of deer specimens. Eleven of the ELISA-positives were also strongly positive in immunoblot analyses. The peripheral blood DNA of seroreactive animals was tested by semi-nested PCR. The specific 275 bp fragment of the pol gene was amplified only in one sample collected from a red deer and the analysis of its sequence showed the highest homology for European BFV isolates. Such results may suggest the existence of a new FV reservoir in bison as well as in deer populations. Whether the origin of such infections stems from a new FV or is the result of BFV inter-species transmission remains to be clarified.

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)

Orthobunyavirus spike architecture and recognition by neutralizing antibodies.

Orthobunyaviruses (OBVs) form a distinct genus of arthropod-borne bunyaviruses that can cause severe disease upon zoonotic transmission to humans. Antigenic drift or genome segment re-assortment have in the past resulted in new pathogenic OBVs, making them potential candidates for causing emerging zoonoses in the future. Low-resolution electron cryo-tomography studies have shown that OBV particles feature prominent trimeric spikes, but their molecular organization remained unknown. Here we report X-ray crystallography studies of four different OBVs showing that the spikes are formed by an N-terminal extension of the fusion glycoprotein Gc. Using Schmallenberg virus, a recently emerged OBV, we also show that the projecting spike is the major target of the neutralizing antibody response, and provide X-ray structures in complex with two protecting antibodies. We further show that immunization of mice with the spike domains elicits virtually sterilizing immunity, providing fundamental knowledge essential in the preparation for potential newly emerging OBV zoonoses.

Sodium dodecyl sulfate as a viral inactivator and future perspectives in the control of small ruminant lentiviruses / Emprego do dodecil sulfato de sódio como inativador viral e suas perspectivas no controle de lentivírus de pequenos ruminantes

Infections by small ruminant lentiviruses (SRLVs) affect goats and sheep causing chronic multisystemic diseases that generate great economic losses. The caprine lentivirus (CLV) and the ovine lentivirus (OLV) present tropism for cells of the monocyte/macrophage lineage, which are directly associated with the main route of transmission through the ingestion of milk and colostrum from infected animals. In this manner, controlling this route is of paramount importance. Currently, researches have investigated the use of chemical additives in milk that can preserve colostrum or milk and inactivate microbiological agents. Among the compounds, sodium dodecyl sulfate (SDS) has been shown to be satisfactory in the chemical inactivation of HIV and CLV in milk, and also as a biocide in goat colostrum.(AU)

As lentiviroses de pequenos ruminantes (LVPRs) são infecções que afetam caprinos e ovinos, causando doenças multissistêmicas crônicas, ocasionando grandes perdas econômicas. Os agentes causadores, lentivírus caprino (LVC) e o lentivírus ovino (LVO), apresentam tropismo por células da linhagem monocítico--fagocitária, as quais estão diretamente associadas à principal via de transmissão, por meio da ingestão de leite e colostro provindos de animais infectados. Desse modo, o controle por esta via é de suma importância. Atualmente, pesquisas vêm sendo desenvolvidas para o uso de aditivos químicos no leite, que possam conservar o colostro ou leite, e inativar agentes microbiológicos presentes. Dentre estes, o dodecil sulfato de sódio (SDS) vem apresentando resultados satisfatórios na inativação química do HIV e LVC em leite, e ainda como biocida em colostro caprino.(AU)

Serosurvey of bluetongue, caprine arthritis-encephalitis (CAE) and Maedi-Visna in Barbary sheep (Ammotragus lervia) of a southern Brazilian zoo / Estudo sorológico dos vírus da língua azul, da artrite-encefalite caprina e Maedi-Visna em aoudads (Ammotragus lervia) em um zoológico do Sul do Brasil

Bluetongue (BT) is an infectious and non-contagious disease of compulsory notification which may affect domestic and wild ruminants, transmitted by Culicoides spp. midges. Despite the high morbidity and mortality in sheep, role of wild animals in the BT cycle remains unclear. Caprine arthritis-encephalitis (CAE) and Maedi-Visna virus (MVV) have been reportedly found in goats and sheep, but not described in wildlife species. Accordingly, serum samples from 17 captive Barbary sheep (Ammotragus lervia) from Curitiba zoo, southern Brazil, were tested for bluetongue, caprine arthritis-encephalitis (CAE) and Maedi-Visna viruses by agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA). Antibodies for bluetongue were observed in 6/17 (35.3%) Barbary sheep by AGID test and in 7/17 (41.2%) by ELISA. All samples were negative for the presence of antibodies against caprine arthritis-encephalitis (CAE) and Maedi-Visna viruses. These findings indicate that Barbary sheep may be infected by bluetongue virus and act as wildlife reservoir in both captive and free-range environments.(AU)

A língua azul é uma doença infecciosa e não contagiosa, de notificação obrigatória, que pode afetar ruminantes domésticos e silvestres, transmitida por mosquitos do gênero Culicoides spp. Apesar da alta morbidade e mortalidade em ovelhas, o papel de animais silvestres no ciclo do vírus da língua azul é desconhecido. A artrite encefalite caprina (CAE) e Maedi-visna vírus (MVV) tem sido encontrados em cabras e ovelhas, porém não há descrição em espécies selvagens. Amostras de soro de 17 aoudads (Ammotragus lervia), mantidos em cativeiro no Zoológico de Curitiba, Sul do Brasil, foram testadas para os vírus da língua azul, da artrite encefalite caprina (CAE) e Maedi-visna, utilizando imunodifusão em gel de ágar e o teste de ELISA (enzyme linked immunosorbent assay). Foram observados anticorpos para o vírus da língua azul em 35,3% (6/17) aoudads utilizando a imunodifusão em gel de ágar e 41,2% (7/17) no ELISA. Todas as amostras foram negativas para a presença de anticorpos contra os vírus da artrite encefalite caprina e Maedi-visna. Esses resultados indicam que os aoudads podem ser infectados pelo vírus da língua azul e atuar como um reservatório silvestre tanto em cativeiro quanto em vida livre.(AU)

Ocorrência e fatores de risco associados à infecção por lentivírus de pequenos ruminantes no Estado de Sergipe / Occorrence and risk factors associated with small ruminant lentivirus infection in the State of Sergipe, Brazil

Os lentiviros de pequenos ruminantes (LVPR) são responsáveis por enfermidades infecciosas e multissistêmicas causadas pelo Vírus da Artrite Encefalite Caprina (CAEV) e o Vírus da Maedi-Visna (MVV), e se apresentam sob as formas clínicas: articular, mamária, respiratória e nervosa. Desta forma esse trabalho objetivou determinar a ocorrência e avaliar os fatores de risco associados à infecção por LVPR no Estado de Sergipe, Brasil. Foram coletadas amostras sanguíneas de 1200 ovinos e 675 caprinos oriundos respectivamente de 60 e 41 propriedades localizadas em 25 municípios sergipanos no período de 2011 a 2014. Os diagnósticos dos LVPR foram determinados pela técnica sorológica de Imunodifusão em Gel Ágar (IDGA) usando o kit comercial da marca Biovetech®. Os dados das variáveis associadas aos fatores de risco foram obtidos a partir de questionários aplicados aos proprietários dos rebanhos e analisados estatisticamente. As frequências absolutas e relativas foram determinadas por análise estatística descritiva e os fatores de risco por análise univariada das variáveis de interesse pelo Teste de Qui-quadrado de Pearson e Exato de Fisher, quando necessário, e em seguida submetidos à análise de regressão logística. Foi evidenciada uma soropositividade de 5,03% (34/675) em caprinos e 1,50% em ovinos com 26,82% (11/41) e 28,33% (17/60) das propriedades apresentando ao menos um animal positivo respectivamente. Na análise dos fatores de risco, não foram observadas diferenças significantes para os ovinos, enquanto que, para os caprinos, rebanhos acima de 100 animais, que pastejam em áreas comuns com outros rebanhos, em uma distância ≤500 metros entre as propriedades, que adotam medidas biotecnológicas da reprodução e não utilizam agulhas estéreis, são mais susceptíveis à infecção por LVPR. Sendo assim, conclui-se que, há a presença dos LVPR em rebanhos sergipanos, e mesmo que em baixas frequências faz-se necessário a implementação de medidas profiláticas devido a possibilidade de expansão e desenvolvimento da caprinocultura do estado, e o alto padrão genético da raça Santa Inês.(AU)

The lentiviruses of small ruminants are infectious and multisystemic diseases caused by the Caprine Arthritis Encephalitis Virus (CAEV) and the Maedi-Visna Virus (MVV), and present the clinical forms: articular, mammary, respiratory and nervous. This work aimed to determine the occurrence and to evaluate the risk factors associated with lentivirus infection of small ruminants in the State of Sergipe, Brazil. Blood samples were collected from 1200 sheep and 675 goats from 60 and 41 farms respectively, located in 25 Sergipe municipalities from 2011 to 2014. The diagnosis of small ruminant lentiviruses (LVPR) was determined by the serological technique of Immunodiffusion in Gel Agar (IDGA) using the commercial kit of the brand Biovetech®. Data from the variables associated with risk factors were obtained from questionnaires applied to the owners of the herds and analyzed statistically. Absolute and relative frequencies were determined by descriptive statistical analysis and risk factors by univariate analysis of the variables of interest by Pearson's Chi-square test and Fisher's exact test, when necessary. A logistic regression analysis was used, considering as a dependent variable for LVPR infection the reactive or non-reactive result observed in the IDGA. A seropositivity of 5.03% (34/675) was observed in goats and 1.50% in sheep with 26.82% (11/41) and 28.33% (17/60) of the properties had at least one animal positive respectively. The analysis of the risk factors, no significant differences were observed for sheep, while for goats, herds above 100 animals grazing in common areas with other herds, at a distance ≤ 500 meters between the properties, that adopt Biotechnological measures of reproduction and do not use sterile needles, are more susceptible to LVPR infection. Therefore, it´s concluded there is presence of lentiviruses of small ruminants in sergipan herds, and even if at low frequencies it is necessary to implement prophylactic measures due to the possibility of expansion and development of goat breeding of the state and the high genetic standard of the Santa Inês breed.(AU)

Papillomaviruses in ruminants: An update.

Papillomaviruses (PVs) are complex viruses which infect the skin or mucosae of a broad range of amniotes worldwide. They cause benign or malignant lesions depending on environmental factors, virus oncogenicity and the location of infection. Bovine papillomaviruses (BPVs) are the second most studied PVs beyond human PVs. In the past few years, genetic characterization of animal PVs has increased due to the availability of new techniques, which simplified the sequencing of entire genomes. Therefore, this review aims to provide an update of the current epidemiology, classification and genome features of ruminant PVs (mainly BPVs) affecting animals worldwide. The review also aimed to clarify the key differences between the high-risk Delta papillomaviruses and the seemingly low-risk Xi, Epsilon, Dyoxi and Dyokappapillomavirus as well as the recently described PVs BPV18, 19, 21 and PpuPV1 that belongs to an unclassified genus.

Epidemiological survey in single-species flocks from Poland reveals expanded genetic and antigenic diversity of small ruminant lentiviruses.

Small ruminant lentivirus (SRLV) infections are widespread in Poland and circulation of subtypes A1, A12, A13, B1 and B2 was detected. The present work aimed at extending previous study based on the analysis of a larger number of animals from single-species flocks. Animals were selected for genetic analysis based on serological reactivity towards a range of recombinant antigens derived from Gag and Env viral proteins. Phylogenetic analysis revealed the existence of subtypes B2 and A12 in both goats and sheep and subtypes A1 and B1 in goats only. In addition, two novel subtypes, A16 and A17, were found in goats. Co-infections with strains belonging to different subtypes within A and B groups were detected in 1 sheep and 4 goats originating from four flocks. Although the reactivity of serum samples towards the recombinant antigens confirmed immunological relatedness between Gag epitopes of different subtypes and the cross-reactive nature of Gag antibodies, eleven serum samples failed to react with antigens representing all subtypes detected up-to-date in Poland, highlighting the limitations of the serological diagnosis. These data showed the complex nature of SRLV subtypes circulating in sheep and goats in Poland and the need for improving SRLV-related diagnostic capacity.

Orf virus infection in Alaskan mountain goats, Dall's sheep, muskoxen, caribou and Sitka black-tailed deer.

BACKGROUND: The zoonotic Orf virus (ORFV; genus Parapoxvirus, Poxviridae family) occurs worldwide and is transmitted between sheep and goats, wildlife and man. Archived tissue samples from 16 Alaskan wildlife cases, representing mountain goat (Oreamnos americanus, n = 8), Dall's sheep (Ovis dalli dalli, n = 3), muskox (Ovibos moschatus, n = 3), Sitka black-tailed deer (Odocoileus hemionus sitkensis, n = 1) and caribou (Rangifer tarandus granti, n = 1), were analyzed. RESULTS: Clinical signs and pathology were most severe in mountain goats, affecting most mucocutaneous regions, including palpebrae, nares, lips, anus, prepuce or vulva, as well as coronary bands. The proliferative masses were solid and nodular, covered by dark friable crusts. For Dall's sheep lambs and juveniles, the gross lesions were similar to those of mountain goats, but not as extensive. The muskoxen displayed ulcerative lesions on the legs. The caribou had two ulcerative lesions on the upper lip, as well as lesions on the distal part of the legs, around the main and dew claws. A large hairless spherical mass, with the characteristics of a fibroma, was sampled from a Sitka black-tailed deer, which did not show proliferative lesions typical of an ORFV infection. Polymerase chain reaction analyses for B2L, GIF, vIL-10 and ATI demonstrated ORFV specific DNA in all cases. Sequences from Dall's sheep formed a separate cluster, comparable to ORFV from domestic sheep. Sequences from the other species were different from the Dall's sheep sequences, but almost identical to each other. CONCLUSIONS: This is the first major investigation of parapoxvirus infections in large Alaskan game species, and the first report of parapoxvirus infection in caribou and Sitka black-tailed deer. This study shows that most of the wild ruminant species in Alaska and from most parts of Alaska, can carry and be affected by ORFV. These findings call for attention to transmission of ORFV from wildlife to livestock and to hunters, subsistence harvesters, and wildlife biologists.

Gammaherpesvirus (Type 1 Ruminant Rhadinovirus) in Muskox ( Ovibos moschatus) in Greenland.

The world's native distribution of muskox ( Ovibos moschatus) is restricted to Canada and Greenland, and a muskox-specific gammaherpesvirus has been described from Canadian populations. We analyzed spleen samples from the Kangerlussuaq muskox population in Greenland and identified muskox gammaherpes by PCR and sequencing.

Teratogenic bluetongue and related orbivirus infections in pregnant ruminant livestock: timing and pathogen genetics are critical.

Congenital infections of domestic animals with viruses in several families, including Bunyaviridae, Flaviridae, Parvoviridae, and Reoviridae, are the cause of naturally occurring teratogenic central nervous system and/or musculoskeletal defects (arthrogryposis) in domestic animals. Congenital infections of ruminant livestock with bluetongue virus (BTV) and some related members of the genus Orbivirus (family Reoviridae) have clearly shown the critical role of gestational age at infection in determining outcome. Specifically, fetuses infected prior to mid-gestation that survive congenital BTV infection are born with cavitating central nervous system defects that range from severe hydranencephaly to cerebral cysts (porencephaly). Generally, the younger the fetus (in terms of gestational age) at infection, the more severe the teratogenic lesion at birth. Age-dependent virus infection and destruction of neuronal and/or glial cell precursors that populate the developing central nervous system are responsible for these naturally occurring virus-induced congenital defects of animals, thus lesions are most severe when progenitor cells are infected prior to their normal migration during embryogenesis. Whereas congenital infection is characteristic of certain BTV strains, notably live-attenuated (modified-live) vaccine viruses that have been passaged in embryonating eggs, transplacental transmission is not characteristic of many field strains of the virus and much remains to be determined regarding the genetic determinants of transplacental transmission of individual virus strains.

Transplacental transmission of Neospora caninum in naturally infected small ruminants from northeastern Brazil / Transmissão transplacentária de Neospora caninum em pequenos ruminantes infectados naturalmente do Nordeste do Brasil

Toxoplasma gondii and Neospora caninum are causative agents of abortion in sheep and goats. Thus, the present study aimed to describe the transplacental transmission of these protozoans in small ruminants of northeastern Brazil. Seventeen fetuses (6 goats and 11 sheep) from farms with history of abortion were necropsied and samples were collected from different tissues (brain, liver, lung, kidney and heart). The samples were analyzed by PCR, histopathology (HP) and immunohistochemistry (IHC) to evaluate whether T. gondii and/or N. caninum infection were the cause of abortion. None of the samples was positive for T. gondii according to PCR and IHC results. Some brain, liver, lung, kidney and heart samples of goat fetuses were positive for N. caninum by PCR. In the histopathology, mild mononuclear infiltration and necrosis with calcification were observed in the liver and brain of one goat fetus, respectively, that also was positive for N. caninum by PCR and IHC. The results confirmed vertical transmission of N. caninum in naturally infected goats of northeastern, Brazil.(AU)

Toxoplasma gondii e Neospora caninum são reconhecidos como protozoário causadores de aborto em ovinos e caprinos. Desta forma, objetivou-se descrever a transmissão transplacentária desses agentes em pequenos ruminantes na região Nordeste do Brasil. Foram examinados seis fetos caprinos e onze fetos ovinos, totalizando 78 amostras de diferentes tecidos (cérebro, fígado, pulmão, rim e coração) provenientes de propriedades rurais com histórico de aborto. As amostras foram analisadas por Reação em Cadeia da Polimerase (PCR), Histopatologia (HP) e Imunohistoquímica (IHQ), com a finalidade de associar o abortamento a T. gondii e/ou N. caninum. Nenhuma amostra foi positiva na PCR e IHQ para T. gondii. Algumas amostras de cérebro, fígado, pulmão, rim e coração de fetos de caprinos e ovinos foram positivas na PCR para N. caninum. Na histopatologia foi observado leve infiltrado mononuclear no fígado e necrose com calcificação no SNC de um caprino, associada à imunomarcação positiva para N. caninum na IHQ e PCR positiva. Os resultados confirmam a transmissão vertical de N. caninum em caprinos naturalmente infectados na região nordeste do Brasil.(AU)

Competitive enzyme-linked immunosorbent assay using baculovirus-expressed VP7 for detection of epizootic haemorrhagic disease virus (EHDV) antibodies.

Epizootic haemorrhagic disease (EHD) is a vector-borne infectious viral disease of domestic and wild ruminants. EHD could spread from infected northern African countries in free territories like the EU; therefore, the availability of diagnostic assays would represent key components for adequate surveillance and control programs. In this study, the gene encoding the VP7 protein of EHD virus (EHDV) was expressed into a baculovirus-infected insect cell system. With this unpurified protein we developed a home-made competitive ELISA (cELISA) and a total number of 275 serum samples, originating from domestic and wild ruminants, were tested. 74/275 were previously shown to be positive for EHDV antibodies by a commercially available ELISA kit. A "very good" agreement was demonstrated when compared to a commercial ELISA kit (Cohen's kappa value=0.832). Samples which caused disagreement between the two assays originated from wildlife which highlights the need for further validation by using serum samples from wild animals.

Whole-genome sequences of Odocoileus hemionus deer adenovirus isolates from deer, moose and elk are highly conserved and support a new species in the genus Atadenovirus.

We present the first complete genome sequence of Odocoileus hemionus deer adenovirus 1 (OdAdV-1). This virus can cause sporadic haemorrhagic disease in cervids, although epizootics with high mortality have occurred in California. OdAdV-1 has been placed in the genus Atadenovirus, based on partial hexon, pVIII and fibre genes. Ten field isolates recovered from naturally infected mule deer (Odocoileus hemionus), white-tailed deer (Odocoileus virginiana) and moose (Alces alces) from Wyoming, black-tailed deer (Odocoileus hemionus columbianus) from California, and Rocky Mountain elk (Cervus elaphus nelsoni) from Colorado and Washington state were sequenced. The genome lengths ranged from 30 620 to 30 699 bp, contained the predicted proteins and gene organization typical of members of genus Atadenovirus, and had a high percentage of A/T nucleotides (66.7 %). Phylogenic analysis found that the closest ancestry was with ruminant atadenoviruses, while a divergence of the hexon, polymerase and penton base proteins of more than 15 % supports classification as a new species. Genetic global comparison between the 10 isolates found an overall 99 % identity, but greater divergence was found between those recovered from moose and elk as compared to deer, and a single variable region contained most of these differences. Our findings demonstrate that OdAdV-1 is highly conserved between 10 isolates recovered from multiple related cervid species, but genotypic differences, largely localized to a variable region, define two strains. We propose that the virus type name be changed to cervid adenovirus 1, with the species name Cervid atadenovirus A. Sequence data were used to develop molecular assays for improved detection and genotyping.

Investigação sorológica de Rickettsia rickettsii e Coxiella burnetii em caprinos e ovinos no entorno do Parque Nacional da Serra das Confusões, Piauí / Infection survey of Rickettsia rickettsii and Coxiella burnetii in sheep and goats from National Park of Serra das Confusões, Piauí

As doenças causadas por bactérias dos gêneros Rickettsia e Coxiella possuem como vetores artrópodes hematófagos, na sua maioria carrapatos, que atuam diretamente na transmissão de patógenos responsáveis por enfermidades de grande impacto na medicina veterinária e humana. O presente estudo objetivou realizar uma investigação sorológica de Rickettsia rickettsii e Coxiella burnetii em caprinos e ovinos criados no entorno do Parque Nacional da Serra das Confusões (PNSC), localizado no estado do Piauí, região nordeste do Brasil. Amostras de soro de 202 caprinos e 153 ovinos foram testadas pela Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos anti-R. rickettsii e anti-C. burnetii, sendo consideradas positivas quando apresentaram títulos ≥ 64. Carrapatos em fase de parasitismo foram coletados e identificados. Todas as amostras de caprinos e ovinos foram soronegativas para antígenos de R. rickettsii. Foi verificado soropositividade em 2% (3/153) das amostras de ovinos para C. burnetii, com títulos variando de 64 a 4.096. As amostras de caprinos não foram reagentes ao referido antígeno. Não foi observado parasitismo em caprinos por carrapatos. No total, foram coletados 56 carrapatos parasitando 15 ovinos (9,8%), todos identificados como Rhipicephalus microplus. O estudo demonstrou a ausência de anticorpos anti-R. rickettsii nas amostras de caprinos e ovinos, ausência de anticorpos anti-C. burnetii em caprinos; e possibilitou o primeiro relato da ocorrência sorológica de C. burnetii em ovinos nesta região do Brasil.(AU)

The diseases caused by bacteria from the genera Rickettsia and Coxiella have hematophagous arthropods as vectors, mostly by ticks, which act directly on the transmission of pathogens that are responsible for diseases with major impact on veterinary and human medicine. The present study aimed to survey the infection of Rickettsia rickettsii and Coxiella burnetii in sheep and goats surrounding in the National Park of Serra das Confusões (NPSC), located in the state of Piauí, Northeast of Brazil. Serum samples from 202 goats and 153 sheep were tested by Indirect Immunofluorescence Assay (IFA) for the detection of antibodies against R. rickettsii and C. burnetii. The samples were considered positive when they showed titers ≥ 64. Ticks in parasitic stage were collected and identified. All samples from sheep and goats were seronegative for R. rickettsii. Seropositivity was verified in 2% (3/153) of the samples of sheep for C. burnetii, with titers ranging from 64 to 4096. The serum samples obtained from goats were seronegative to the above antigens. In total, 56 ticks were collected from 15 sheep (9.8%) all identified as Rhipicephalus microplus. The study demonstrated absence of infection by R. rickettsii in samples of sheep and goats, absence of infection of C. burnetii in goats; and the first report of serological occurrence of C. burnetii in sheep in this region of Brazil.(AU)

Caprine lentivirus in sheep milk and semen / Lentivírus caprino em leite e sêmen de ovinos

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.(AU)

Com o objetivo de detectar a presença do lentivírus caprino (LVC) no leite de ovelhas e no sêmen 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 lactação, ocorrendo cinco coletas por animal, totalizando 50 amostras. Quanto aos carneiros, realizaram-se oito coletas de sêmen por animal, durante um ano de experimentação, totalizando 32 amostras. As amostras de leite e de sêmen foram submetidas à extração de DNA e à prova de reação em cadeia da polimerase do tipo nested (nPCR) visando à detecção de DNA proviral do LVC. Oito (16%) amostras de leite foram positivas na nPCR oriundas de duas ovelhas. Apenas uma (3,12%) amostra de sêmen apresentou positividade. Produtos da amplificação foram sequenciados, confirmando-se tratar de sequência genômica do LVC. Dessa forma, demonstrou-se a presença do DNA proviral do LVC em leite e sêmen de ovinos, confirmando a viabilidade da infecção entre espécies e, assim, alertando sobre o risco de que a infecção seja disseminada.(AU)