The complete genome of equid herpesvirus-1 (EHV-1) field isolates from Argentina reveals an interspecific recombinant strain.

The Equid alphaherpesvirus type 1 (EHV-1) infection can have devastating economic consequences in the horse industry due to large-scale outbreaks of abortions, perinatal foal mortality, and myeloencephalopathy. The present study analyzed the genome of two isolates obtained from aborted fetuses in Argentina, E/745/99 and E/1297/07. The E745/99 genome shares 98.2% sequence identity with Ab4, a reference EHV-1 strain. The E/1297/07 genome shares 99.8% identity with NY03, a recombinant strain containing part of ORF64 and part of the intergenic region from Equid alphaherpesvirus-4 (EHV-4). The E/1297/07 genome has the same breakpoints as other United States and Japanese recombinants, including NY03. The recombinant regions have varying numbers of tandem repeat sequences and different minor parental sequences (EHV-4), suggesting distinct origins of the recombinant events. These are the first complete genomes of EHV-1 from Argentina and South America available in the Databases.

Mapping Eastern (EEE) and Venezuelan Equine Encephalitides (VEE) among Equines Using Geographical Information Systems, Colombia, 2008-2019.

INTRODUCTION: Eastern equine encephalitis virus (EEEV) and Venezuelan equine encephalitis virus (VEEV) viruses are zoonotic pathogens affecting humans, particularly equines. These neuroarboviruses compromise the central nervous system and can be fatal in different hosts. Both have significantly influenced Colombia; however, few studies analyse its behaviour, and none develop maps using geographic information systems to characterise it. OBJECTIVE: To describe the temporal-spatial distribution of those viruses in Colombia between 2008 and 2019. METHODS: Retrospective cross-sectional descriptive study, based on weekly reports by municipalities of the ICA, of the surveillance of both arboviruses in equines, in Colombia, from 2008 to 2019. The data were converted into databases in Microsoft Access 365®, and multiple epidemiological maps were generated with the Kosmo RC1®3.0 software coupled to shape files of all municipalities in the country. RESULTS: In the study period, 96 cases of EEE and 70 of VEE were reported, with 58% of EEE cases occurring in 2016 and 20% of EEV cases in 2013. The most affected municipalities for EEE corresponded to the department of Casanare: Yopal (20), Aguazul (16), and Tauramena (10). In total, 40 municipalities in the country reported ≥1 case of EEE. CONCLUSIONS: The maps allow a quick appreciation of groups of neighbouring municipalities in different departments (1° political division) and regions of the country affected by those viruses, which helps consider the expansion of the disease associated with mobility and transport of equines between other municipalities, also including international borders, such as is the case with Venezuela. In that country, especially for EEV, municipalities in the department of Cesar are bordering and at risk for that arboviral infection. there is a high risk of equine encephalitis outbreaks, especially for VEE. This poses a risk also, for municipalities in the department of Cesar, bordering with Venezuela.

Nodular lymphocytic conjunctivitis in a horse - case report / Conjuntivite linfocítica nodular em equino - relato de caso

The case of an equine with nodular lymphocytic conjunctivitis is described. A 14-year-old crossbred mare was seen presenting with development of a mass in the nasal angle of the right eye, behind the third eyelid. The tutor reported slow growth over 4 years, always accompanied by epiphora, and that no treatment had been performed prior to consultation. The histopathological and immunohistochemical results found a nodular, subepithelial structure, composed predominantly of densely packed small lymphocytes. Through the exams, associated with studies with monoclonal anti B lymphocyte antibodies and polyclonal anti T lymphocyte antibodies, the diagnosis of nodular lymphocytic conjunctivitis was reached. Only clinical pharmacological treatment was chosen, based on the use of topical and intralesional hydrocortisone acetate. After one month of treatment the mass completely disappeared without sequelae.

Descreve-se o caso de um equino com conjuntivite linfocítica nodular. Uma égua, mestiça, de 14 anos, foi atendida apresentando desenvolvimento de uma massa no ângulo nasal do olho direito, atrás da terceira pálpebra. O tutor relatou um crescimento lento durante 4 anos, sempre acompanhado de epífora, para o qual não foi realizado tratamento prévio à consulta. Os resultados histopatológico e imuno-histoquímico constataram uma estrutura nodular e subepitelial, composta predominantemente por linfocitos pequenos densamente agrupados. Por meio dos exames, associados a estudos com anticorpos monoclonais antilinfocitos B e anticorpos policlonais antilinfocitos T, chegou-se ao diagnóstico de conjuntivite linfocítica nodular. Optou-se apenas pelo tratamento clínico farmacológico, baseando-se na utilização de acetato de hidrocortisona tópica e intralesional. Após um mês de tratamento, a massa involuiu por completo, sem deixar sequelas.

Equine herpesvirus myeloencephalopathy

Clinical History: A euthanized, 14-year-old, Warmblood gelding that had participated in an equine show was presented for necropsy and diagnostic workup to the San Bernardino laboratory of the California Animal Health and Food Safety Laboratory System. The animal had a 4-day history of mildly swollen limbs and hyperthermia, and developed neurologic signs shortly before euthanasia. Necropsy Findings: The urinary bladder had multifocal to coalescing hemorrhages in the mucosa and approximately 5 ml of turbid urine with sandy sludge (Figure 1). The entire spinal cord was removed and cross-sectioned serially after fixation in 10% neutral-buffered formalin during 48h. Multifocally, in multiple sections of the cervical, thoracic, and lumbar segments, there were uni- or bilateral and asymmetrical, wedge-shaped areas of gray discoloration and hemorrhage (Figures 2-4). In addition, there were extensive hemorrhages around the nerve roots of the cauda equina. Follow-up questions: Five differential diagnoses for the gross lesions in the spinal cord (Fig. 1B-1D) Microscopic description for the lesions in the spinal cord (Fig. 2A-2D) Most likely cause based on clinical history and gross and microscopic findings.

Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São Paulo / West Nile Fever virus infection in horses in São Paulo state, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses andbirds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmedin a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse withnervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus(neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case reportis to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (SãoPaulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and arealso routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow,the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering,it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelviclimb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervoussystem samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negativeresults. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase...(AU)

Infecção pelo vírus da Febre do Nilo Ocidental em equinos no Estado de São Paulo / West Nile Fever virus infection in horses in São Paulo state, Brazil

Background: The West Nile virus (WNV) antibodies were reported in Brazil in the serum samples taken from horses andbirds in the Midwest region and Paraíba state in 2008 and 2013, respectively. In 2014, the first human case was confirmedin a rural worker in the state of Piauí and, in 2018, the virus was isolated from the central nervous system of a horse withnervous symptoms in the state of Espírito Santo. The virus is a member of the Flaviviridae family of the genus Flavivirus(neurotropic), infecting several mammalian species, with humans and horses being the most susceptible. Approximately35% of horses develop clinical signs, thus they are considered the best sentinels for this disease. The aim of this case reportis to describe the first confirmed cases of West Nile Fever (WNF) in two horses in the state of São Paulo.Cases: Two horses with neurological symptoms were treated at the Veterinary Hospital of Cruzeiro do Sul University (SãoPaulo, SP), in 2019. Both horses came from neighboring regions that have a large Atlantic Forest preservation area and arealso routes for migratory birds, known to be competent hosts for transmitting the West Nile Fever virus, such as the swallow,the falcon and the hawk. The first one had symptoms, such as weakness and sporadic seizures; however, after recovering,it was hospitalized a few days later due to kidney failure and laminitis. The second one showed incoordination, pelviclimb weakness, and was walking in circles, evolving to seizures. Both animals were euthanized, and their central nervoussystem samples and total blood samples were tested for rabies, herpes virus, and WNV; the first 2 tests showed negativeresults. Ribonucleic acids (RNA) were extracted from erythrocytes using the polymerase...

Seroprevalence and risk factors associated with seropositivity for equine encephalomyelitis virus in horses in Rio Grande do Norte, Brazil

The present objective was to investigate the presence of anti-equine viral encephalomyelitis (EVE) antibodies and the possible risk factors for its dissemination in horses raised in the East and West Potiguar mesoregions of the state of Rio Grande do Norte, Brazil. Serological diagnosis for neutralizing antibodies against Eastern (EEEV), Western (WEEV) and Venezuelan (VEEV). Equine viral encephalomyelitis was performed using a seroneutralization technique on 811 blood samples from horses from ninety properties and sixteen municipalities between July 2018 and February 2019. Factors associated with EVE were evaluated using an investigative epidemiological questionnaire, and the data were statistically analyzed using the Epi Info 3.5.2 software with a confidence level of 95%. The seroprevalence of anti-EVE antibodies was 14.2% (115), with 10.36% (84) for EEEV, 6.9% (56) for WEEV, and null for EVE. When analyzing risk factors, it can be concluded that horses raised in properties that do not clean installations and/or rent out their pasture are more likely to have anti-EVE antibodies. These results show evidence that horses raised in the East and West Potiguar mesoregions were exposed to EEEV and WEEV, thus reinforcing the importance of vaccination and serological survey of nonvaccinated horses as a means of monitoring the disease.

Fatores associados à infecção pelo vírus da estomatite vesicular em equídeos no estado do Rio Grande do Norte, Brasil / Factors associated with infection by the vesicular stomatitis virus in equines in the state of Rio Grande do Norte, Brazil

Objetivou-se investigar a presença do Vírus da Estomatite Vesicular (VEV) e seus fatores de risco para ocorrência e disseminação da enfermidade em equídeos das mesorregiões Leste e Oeste Potiguar do estado do Rio Grande do Norte, Brasil. Foram analisadas pela técnica de virusneutralização, 809 amostras sanguíneas de equídeos provenientes de noventa propriedades de dezesseis municípios Potiguares durante os meses de julho de 2018 a fevereiro de 2019. Os fatores de riscos associados ao VEV foram avaliados por meio de questionário epidemiológico e os dados submetidos a análise estatística no programa IBM SPSS Statistics versão 21.0 com nível de confiança de 95%. Posteriormente, todas as variáveis estatisticamente significantes foram submetidas a análise de regressão de Poisson. A soroprevalência de anticorpos anti-VEV foi 24,6% (199/809), sendo 3,2% (13/402) de soropositivos na mesorregião Leste e 45,7% (186/407) na do Oeste Potiguar. Com relação aos sorotipos, observou-se uma prevalência de 3,8% (31/809) e 24,5% (198/809) para Indiana 2 e 3 respectivamente, com 15,1% (30/198) de coinfecção. Equídeos criados na mesorregião Oeste, em propriedades que não realizam quarentena e onde os animais enfermos são mantidos no rebanho, foram consideradas fatores predisponentes a infecção pelo VEV. Esses resultados demonstram a circulação do VEV em equídeos no Rio Grande do Norte, com destaque ao Oeste Potiguar, e sendo necessário a aplicação de medidas sanitárias que impeçam introdução e disseminação do vírus ente as espécies susceptíveis, principalmente em condições climáticas favoráveis para a sua manutenção, no ambiente de criação e pastagens.

This study aimed to investigate the presence of Vesicular stomatitis virus (VSV) and risk factors for its occurrence and dissemination in equines from the Eastern and Western mesoregions of the state of Rio Grande do Norte, Brazil. Blood samples were analyzed, by Serum Virus Neutralization Assay, from 809 animals belonging to 90 properties distributed in sixteen municipalities from July 2018 to February 2019. Risk factors were assessed using an epidemiological questionnaire. Data were submitted to statistical analysis using the software IBM SPSS Statistics, version 21.0 with a 95% confidence level. Also, all statistically significant variables were subjected to Poisson regression analysis. The occurrence of anti-VSV antibodies was 24.6% (199/809) with 3.2% (13/402) and 45.7% (186/407) of seropositivity in the Western and Eastern mesoregion, respectively. Regarding serotypes, there were an occurrence of 3.8% (31/809) and 24.5% (198/809) for Indiana 2 and 3, respectively, and 15.1% (30/198) of co-infection for both. Equines kept of the Western mesoregion, on properties that do not quarantine, and where sick animals are kept in the herd, were considered risk factors for LVV infection. These results demonstrate the presence of VSV in equines in Rio Grande do Norte, with emphasis on Oeste Potiguar, and that sanitary measures must be adopted to prevent the introduction and viral spreading among susceptible species, especially due to favorable climatic conditions for the maintenance of VSV in the breeding and pasture environment.

Retrospective serological survey for influenza in horses from Brazil.

Equine influenza (EI) virus is one of the most economically important pathogens of respiratory diseases of horses worldwide. Despite availability of vaccines for control of EI, the highly contagious nature and variability properties of the virus mean global outbreaks occur. Thus, continuous surveillance programs, including seroprevalence studies of disease in different countries, may contribute to better control of the disease. In this study, the seroprevalence of equine influenza in 850 horses from Brazil was investigated. The serodiagnosis was based on the single radial hemolysis (SRH) assay using influenza A/equine/Richmond/1/2007 (H3N8) antigen. Antibodies against A/equine/Richmond/1/07 (H3N8) were detected in 44.7% (380/850, 95% CI: 41.4-48.1%) of horses. Seroprevalence was significantly lower (p = 0.001) in younger animals (< 5 years, 38.6%) than in "adult" animals (5-14 years, 52.1%). There was also a significant relationship between the year of sampling and seroprevalence (p < 0.0005). The mean SRH antibody value was 42.0 mm2 (range 4-238.9 mm2), with the majority of horses (95.3%) having an SRH value ≤ 150 mm2, which is considered an insufficient level for protection of equine hosts against influenza infections and potential virus shedding. These findings indicate the need to reinforce preventive/control measures against equine influenza in Brazil.

Occurrence of Leptospira spp. and factors associated with the infection in horses from a military contingent in the Rio Grande do Sul State, Brazil / Ocorrência de Leptospira spp. e fatores associados à infecção em equinos de contingente militar no estado do Rio Grande do Sul, Brasil

The seroprevalence of Leptospira spp. and infection risk factors were investigated in horses from a military contingent in the Rio Grande do Sul State, Brazil. A total of 446 horses were evaluated and categorized into 309 mares, 11 stallions, and 126 horses with an average age of three years. To determine seroprevalence, serum samples from all horses were submitted to the microscopic agglutination test against 12 serovars belonging to nine serogroups, usually circulating in equine populations. To investigate the possible risk factors of infection by Leptospira spp., questionnaires were applied in that military unit. The seroprevalence in the horses with an average age of 3 years, mares, and stallions were 57.94% (73/126), 54.05% (165/309), and 45.45% (5/11), respectively. The main risk factors identified were contact with different water sources, the presence of rodents, and contact with wild and domestic animals. The high seroprevalence of anti­Leptospira spp. antibodies may be associated with the presence of these risk factors. Therefore, the exposure of horses to possible reservoirs of Leptospira spp. should be minimized. Also, the immunoprophylaxis protocol should be reviewed and a shorter interval between vaccinations adopted to control leptospirosis in this herd.(AU)

A soroprevalência de Leptospira spp. e os fatores de risco para infecção foram investigados em cavalos de um contingente militar no Rio Grande do Sul, Brasil. Um total de 446 cavalos foram avaliados e categorizados em 309 éguas, 11 garanhões e 126 cavalos com idade média de três anos. Para determinação da soroprevalência, amostras de soro de todos os equinos foram submetidas ao teste de aglutinação microscópica contra 12 sorovares pertencentes a nove sorogrupos, normalmente circulantes em populações equinas. Para investigar os possíveis fatores de risco da infecção por Leptospira spp., foi aplicado um questionário naquela unidade militar. A soroprevalência nos cavalos com idade média de três anos, éguas e garanhões foi de 57,94% (73/126), 54,05% (165/309) e 45,45% (5/11), respectivamente. Os principais fatores de risco identificados foram o contato com diferentes fontes de água, a presença de roedores e o contato com animais silvestres e domésticos. A alta soroprevalência de anticorpos anti­Leptospira spp. pode estar associada à presença desses fatores de risco. Portanto, a exposição destes equinos aos possíveis reservatórios de Leptospira spp. deve ser minimizada. Além disso, o protocolo de imunoprofilaxia deve ser revisto e, possivelmente, um menor intervalo entre as vacinas deve ser adotado para o controle da leptospirose neste plantel.(AU)

Fatores associados à infecção pelo vírus da estomatite vesicular em equídeos no estado do Rio Grande do Norte, Brasil / Factors associated with infection by the vesicular stomatitis virus in equines in the state of Rio Grande do Norte, Brazil

Objetivou-se investigar a presença do Vírus da Estomatite Vesicular (VEV) e seus fatores de risco para ocorrência e disseminação da enfermidade em equídeos das mesorregiões Leste e Oeste Potiguar do estado do Rio Grande do Norte, Brasil. Foram analisadas pela técnica de virusneutralização, 809 amostras sanguíneas de equídeos provenientes de noventa propriedades de dezesseis municípios Potiguares durante os meses de julho de 2018 a fevereiro de 2019. Os fatores de riscos associados ao VEV foram avaliados por meio de questionário epidemiológico e os dados submetidos a análise estatística no programa IBM SPSS Statistics versão 21.0 com nível de confiança de 95%. Posteriormente, todas as variáveis estatisticamente significantes foram submetidas a análise de regressão de Poisson. A soroprevalência de anticorpos anti-VEV foi 24,6% (199/809), sendo 3,2% (13/402) de soropositivos na mesorregião Leste e 45,7% (186/407) na do Oeste Potiguar. Com relação aos sorotipos, observou-se uma prevalência de 3,8% (31/809) e 24,5% (198/809) para Indiana 2 e 3 respectivamente, com 15,1% (30/198) de coinfecção. Equídeos criados na mesorregião Oeste, em propriedades que não realizam quarentena e onde os animais enfermos são mantidos no rebanho, foram consideradas fatores predisponentes a infecção pelo VEV. Esses resultados demonstram a circulação do VEV em equídeos no Rio Grande do Norte, com destaque ao Oeste Potiguar, e sendo necessário a aplicação de medidas sanitárias que impeçam introdução e disseminação do vírus ente as espécies susceptíveis, principalmente em condições climáticas favoráveis para a sua manutenção, no ambiente de criação e pastagens.(AU)

This study aimed to investigate the presence of Vesicular stomatitis virus (VSV) and risk factors for its occurrence and dissemination in equines from the Eastern and Western mesoregions of the state of Rio Grande do Norte, Brazil. Blood samples were analyzed, by Serum Virus Neutralization Assay, from 809 animals belonging to 90 properties distributed in sixteen municipalities from July 2018 to February 2019. Risk factors were assessed using an epidemiological questionnaire. Data were submitted to statistical analysis using the software IBM SPSS Statistics, version 21.0 with a 95% confidence level. Also, all statistically significant variables were subjected to Poisson regression analysis. The occurrence of anti-VSV antibodies was 24.6% (199/809) with 3.2% (13/402) and 45.7% (186/407) of seropositivity in the Western and Eastern mesoregion, respectively. Regarding serotypes, there were an occurrence of 3.8% (31/809) and 24.5% (198/809) for Indiana 2 and 3, respectively, and 15.1% (30/198) of co-infection for both. Equines kept of the Western mesoregion, on properties that do not quarantine, and where sick animals are kept in the herd, were considered risk factors for LVV infection. These results demonstrate the presence of VSV in equines in Rio Grande do Norte, with emphasis on Oeste Potiguar, and that sanitary measures must be adopted to prevent the introduction and viral spreading among susceptible species, especially due to favorable climatic conditions for the maintenance of VSV in the breeding and pasture environment.(AU)

Occurrence of Leptospira spp. and factors associated with the infection in horses from a military contingent in the Rio Grande do Sul State, Brazil / Ocorrência de Leptospira spp. e fatores associados à infecção em equinos de contingente militar no estado do Rio Grande do Sul, Brasil

The seroprevalence of Leptospira spp. and infection risk factors were investigated in horses from a military contingent in the Rio Grande do Sul State, Brazil. A total of 446 horses were evaluated and categorized into 309 mares, 11 stallions, and 126 horses with an average age of three years. To determine seroprevalence, serum samples from all horses were submitted to the microscopic agglutination test against 12 serovars belonging to nine serogroups, usually circulating in equine populations. To investigate the possible risk factors of infection by Leptospira spp., questionnaires were applied in that military unit. The seroprevalence in the horses with an average age of 3 years, mares, and stallions were 57.94% (73/126), 54.05% (165/309), and 45.45% (5/11), respectively. The main risk factors identified were contact with different water sources, the presence of rodents, and contact with wild and domestic animals. The high seroprevalence of anti­Leptospira spp. antibodies may be associated with the presence of these risk factors. Therefore, the exposure of horses to possible reservoirs of Leptospira spp. should be minimized. Also, the immunoprophylaxis protocol should be reviewed and a shorter interval between vaccinations adopted to control leptospirosis in this herd.(AU)

A soroprevalência de Leptospira spp. e os fatores de risco para infecção foram investigados em cavalos de um contingente militar no Rio Grande do Sul, Brasil. Um total de 446 cavalos foram avaliados e categorizados em 309 éguas, 11 garanhões e 126 cavalos com idade média de três anos. Para determinação da soroprevalência, amostras de soro de todos os equinos foram submetidas ao teste de aglutinação microscópica contra 12 sorovares pertencentes a nove sorogrupos, normalmente circulantes em populações equinas. Para investigar os possíveis fatores de risco da infecção por Leptospira spp., foi aplicado um questionário naquela unidade militar. A soroprevalência nos cavalos com idade média de três anos, éguas e garanhões foi de 57,94% (73/126), 54,05% (165/309) e 45,45% (5/11), respectivamente. Os principais fatores de risco identificados foram o contato com diferentes fontes de água, a presença de roedores e o contato com animais silvestres e domésticos. A alta soroprevalência de anticorpos anti­Leptospira spp. pode estar associada à presença desses fatores de risco. Portanto, a exposição destes equinos aos possíveis reservatórios de Leptospira spp. deve ser minimizada. Além disso, o protocolo de imunoprofilaxia deve ser revisto e, possivelmente, um menor intervalo entre as vacinas deve ser adotado para o controle da leptospirose neste plantel.(AU)

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...(AU)

Expressão e caracterização da glicoproteína D do herpesvírus equídeo 1 em Pichia pastoris / Expression and characterization of equid herpesvirus 1 glycoprotein D in Pichia pastoris

O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)

Expressão e caracterização da glicoproteína D do herpesvírus equídeo 1 em Pichia pastoris / Expression and characterization of equid herpesvirus 1 glycoprotein D in Pichia pastoris

O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)

Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)

Venezuelan equine encephalitis virus: the problem is not over for tropical America.

The equine encephalitis viruses, Venezuelan (VEEV), East (EEEV) and West (WEEV), belong to the genus alphavirus, family Togaviridae and still represent a threat for human and animal public health in the Americas. In both, these infections are characterized by high viremia, rash, fever, encephalitis and death. VEEV encephalitis is similar, clinically, to other arboviral diseases, such as dengue, Zika or chikungunya. Most of the alphaviruses are transmitted between vertebrates and mosquitoes. They are able to replicate in a wide number of hosts, including mammals, birds, reptiles, amphibian and arthropods. The VEEV has enzootic and epizootic transmission cycles. At the enzootic one, enzootic strains (subtype I, serotypes D-F and serotypes II-VI) are continuously circulating between mosquitoes and wild rodents in tropical forests and mangroves of the Americas. The main reseroivrs are wild rodent species of the subfamily Sigmodontinae. However, bats can be also accidental reservoirs of VEEV. In this article, we reviewed the main features, epidemiology, clinical aspects and the current perspectives of the VEEV.

Molecular detection, histopathological analysis, and immunohistochemical characterization of equine infectious anemia virus in naturally infected equids.

Equine infectious anemia (EIA), a disease caused by equine infectious anemia virus (EIAV), is considered an obstacle to the development of the horse industry. There is no treatment or vaccine available for EIA, and its pathogenesis, as well as the immune response against the virus, is not fully understood. Therefore, an immunohistochemistry assay was developed for the detection of viral antigens in tissues of equids naturally infected with EIAV. Sections of organs of six equids from Apodi-RN, Brazil, that tested positive for EIA by serological tests (ELISA and AGID) were fixed in 10% formalin solution and embedded in paraffin. Immunohistochemistry was performed using a polyclonal anti-EIAV antibody. EIAV antigens were observed in red spleen pulp cells and hepatic sinusoids, as well as bronchiolar and alveolar epithelial cells of the lungs and proximal and distal tubules of the kidneys. The presence of EIAV in the spleen and liver was expected due to viral tropism by macrophages, which are abundantly present in these organs. However, EIAV was also found in lung and kidney epithelial cells, indicating that the virus infects cell types other than macrophages. In conclusion, the immunohistochemical assay standardized in this study was able to detect EIAV antigens in spleen, liver, kidney and lung cells from naturally infected EIAV equids. Immunostaining observed in the spleen confirms viral tropism by mononuclear phagocytes; however, the presence of EIAV in lung and kidney epithelial cells indicates that virus may be eliminated in urine and/or oronasal secretions, suggesting new routes for viral excretion.

Seroprevalence for equine infectious anaemia in Equidae seized in the municipality of Petrópolis, State of Rio de Janeiro, Brazil, 2015/2018 / Soroprevalência para anemia infecciosa equina em Equídeos apreendidos no município de Petrópolis, RJ, Brasil, 2015/2018

Equine infectious anaemia (EIA) is a globally distributed disease caused by a lentivirus. EIA is a notifiable disease that should be reported to the Official Veterinary Service (SVO). Outbreaks are controlled by sacrificing the EIA-positive animals. The study analysed the prevalence of EIA in 165 Equidae seized on public roads in the municipality of Petrópolis, Rio de Janeiro, Brazil, from 2015 to 2018. After the seizure and identification of the Equidae, blood was collected to diagnose EIA. A prevalence of 11.8% was observed among the Equidae seized, and the main risk factor was the region of seizure. A kernel-smoothed map showed areas with the highest density of EIA-positive Equidae in the centre and to the east of the study area. In this region, the chance of seizing an equine positive for EIA was greater (OR=3.6) relative to the remaining region. The implementation and maintenance of Equidae seizures and subsequent diagnostic testing for EIA in the municipality of Petrópolis were shown to have great importance for the control and prevention of the disease. Indicating the place of seizure on the identification cards of the animals enabled delineation of the locations in the municipality of Petrópolis, that had a higher risk of EIA-positive animals.(AU)

A anemia infecciosa equina (AIE) é uma doença globalmente distribuída causada por um lentivírus. AIE é uma doença de notificação obrigatória que deve ser relatada ao Serviço Veterinário Oficial (SVO), e os surtos são controlados pelo sacrifício dos animais positivos para AIE. O presente estudo analisou a prevalência de AIE em 165 equídeos apreendidos em vias públicas no município de Petrópolis, estado do Rio de Janeiro, Brasil, no período de 2015 a 2018. Após a apreensão e identificação dos equídeos seu sangue foi coletado para diagnosticar a AIE. Observou-se uma prevalência real de 11,8% entre os equídeos apreendidos, sendo que o principal fator de risco identificado foi a região de apreensão. O Mapa de Kernel mostrou que as áreas com maior densidade de equídeos positivos estavam no centro e leste da área de estudo. Nesta região, houve maior chance (OR = 3,6) de apreender um animal positivo para AIE em relação ao da outra região, composto pelos 34 locais restantes. A implementação e manutenção de apreensão de equídeos e posterior teste de diagnóstico para AIE no município de Petrópolis, RJ, mostrou-se de grande importância para o controle e prevenção da doença. A indicação do local da apreensão nos cartões de identificação dos animais possibilitou um delineamento dos locais no município de Petrópolis, RJ, que apresentavam maior risco de animais positivos para AIE.(AU)

High Genomic Variability in Equine Infectious Anemia Virus Obtained from Naturally Infected Horses in Pantanal, Brazil: An Endemic Region Case.

Equine infectious anemia virus (EIAV) is a persistent lentivirus that causes equine infectious anemia (EIA). In Brazil, EIAV is endemic in the Pantanal region, and euthanasia is not mandatory in this area. All of the complete genomic sequences from field viruses are from North America, Asia, and Europe, and only proviral genomic sequences are available. Sequences from Brazilian EIAV are currently available only for gag and LTR regions. Thus, the present study aimed for the first time to sequence the entire EIAV genomic RNA in naturally infected horses from an endemic area in Brazil. RNA in plasma from naturally infected horses was used for next-generation sequencing (NGS), and gaps were filled using Sanger sequencing methodology. Complete viral genomes of EIAV from two horses were obtained and annotated (Access Number: MN560970 and MN560971). Putative genes were analyzed and compared with previously described genes, showing conservation in gag and pol genes and high variations in LTR and env sequences. Amino acid changes were identified in the p26 protein, one of the most common targets used for diagnosis, and p26 molecular modelling showed surface amino acid alterations in some epitopes. Brazilian genome sequences presented 88.6% nucleotide identity with one another and 75.8 to 77.3% with main field strains, such as EIAV Liaoning, Wyoming, Ireland, and Italy isolates. Furthermore, phylogenetic analysis suggested that this Brazilian strain comprises a separate monophyletic group. These results may help to better characterize EIAV and to overcome the challenges of diagnosing and controlling EIA in endemic regions.