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From February to December 2021, 960 serum samples from different climate regions in two Mexican states were analyzed for antibodies against EIAV using competitive enzyme-linked immunosorbent assay (cELISA) and agar gel immunodiffusion (AGID) tests. The overall seroprevalence of EIAV infection in equids determined by ELISA and AGID tests was 19.69% and 17.60%, respectively. The highest rate of seropositivity was observed in the state of Veracruz (23.67%, using AGID test), and the lowest rate was found in the state of Puebla (13.12%, using cELISA test). In total, 17.45% (85/487, using AGID test) and 15.40% (75/487, using cELISA test) of horses were positive for EIAV infection. Donkeys showed a prevalence of 19.87% (94/473) in cELISA and 21.99% (104/476) in AGID. The cELISA was compared with AGID as the gold standard test. The diagnostic sensitivity for horses and donkeys was 90.4% and 92.2%, and the specificity was 97.5% and 97.3%, respectively, with an almost perfect degree of agreement (kappa values of 0.871 and 0.888). Therefore, the overall seroprevalence rate of EIAV infection in equines in the sampled states of Puebla and Veracruz was 19.3%, with 91.4% sensitivity, 97.4% specificity, and an agreement of 96.25%, indicating almost perfect agreement (kappa = 0.880).

De fevereiro a dezembro de 2021, 960 amostras de soro de diferentes regiões climáticas em dois estados mexicanos foram analisadas quanto à presença de anticorpos contra o EIAV, por meio de testes de imunoabsorção enzimática competitiva (cELISA) e imunodifusão em gel de ágar (AGID). A soroprevalência geral da infecção por EIAV em equídeos, determinada pelos testes ELISA e AGID, foi de 19,69% (189 de 960 amostras de soro) e 17,60% (169 de 960 amostras de soro), respectivamente. A maior taxa de soropositividade foi observada no estado de Veracruz (23,67%, usando-se o teste AGID), e a menor taxa foi encontrada no estado de Puebla (13,12%, usando-se o teste cELISA). No total, 17,45% (85/487, usando-se o teste AGID) e 15,40% (75/487, usando-se o teste cELISA) dos cavalos foram positivos para a infecção por EIAV. Os jumentos apresentaram uma prevalência de 19,87% (94/473) no cELISA e 21,99% (104/476) no AGID. O cELISA foi comparado com o AGID como o teste padrão-ouro. A sensibilidade diagnóstica para cavalos e jumentos foi de 90,4% e 92,2%, e a especificidade foi de 97,5% e 97,3%, respectivamente, com um grau de concordância quase perfeito (valores kappa de 0,871 e 0,888). Portanto, a taxa de soroprevalência geral da infecção por EIAV em equinos nos estados de Puebla e Veracruz amostrados foi de 19,3%, com sensibilidade de 91,4%, especificidade de 97,4% e concordância de 96,25%, indicando concordância quase perfeita (kappa = 0,880).

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To assist decision making regarding the National Equine Health Program in the state of Paraná, a study was conducted to estimate the prevalence of infected farms and seropositive animals for Equine Infectious Anemia (EIA) and also identify possible risk factors for the disease. The state was divided into three regions, within which about 300 farms were randomly selected. On the selected farms, a minimum number of animals aged 6 months or older were examined to characterize them as infected or free of EIA. In the sampled farms, an epidemiological questionnaire was applied to investigate possible associations between the disease and its characteristics, including sanitary practices and livestock management. The test used was the Agar Gel Immunodiffusion Test. A total of 2818 equids from 889 farms were tested. In the state, the prevalence of infected farms was 1.55% [0.92; 3.00] and the prevalence of seropositive animals was 0.55% [0.27; 1.00]. Introduction of equids showed an association with EIA (OR=5.5 [1.9; 15.9]). Paraná equine owners should be alerted to the need to observe health precautions regarding EIA when introducing animals to their herd. In 2018, the sensitivity of the Surveillance System for EIA in Paraná was only 1.36%, probably insufficient to change the endemic balance of the disease, therefore, needs to be re-evaluated involving in the process all public and private agents interested in the subject.(AU)

Para auxiliar a tomada de decisões em relação à execução do Programa Nacional de Sanidade dos Equídeos no estado do Paraná, foi realizado um estudo para estimar a prevalência de focos e de animais para Anemia Infecciosa Equina (AIE) e também identificar os possíveis fatores de risco para a doença. O estado foi dividido em três regiões, dentro das quais cerca de 300 propriedades foram aleatoriamente selecionadas. Nas propriedades sorteadas, foi examinado um número mínimo de animais com idade igual ou superior a 6 meses para caracterizá-la como foco (pelo menos um animal soropositivo) ou não foco de AIE. Nas propriedades amostradas foi aplicado um questionário epidemiológico para averiguar as possíveis associações entre a doença e as características da propriedade, incluindo suas práticas sanitárias e zootécnicas. O teste utilizado foi a Imunodifusão em Gel de Ágar (IDGA). Foram testados 2818 equídeos oriundos de 889 propriedades. No estado, a prevalência de focos foi de 1,55% [0,92; 3,00] e a de animais 0,55% [0,27; 1,00]. A introdução de equídeos apresentou associação com a AIE (OR=5,5 [1,9; 15,9]). Os proprietários paranaenses de equídeos devem ser alertados para a necessidade de observar cuidados sanitários relativos à AIE ao introduzir animais nos seus plantéis. Em 2018, a sensibilidade do Sistema de Vigilância para AIE no Paraná foi de apenas 1,36%, provavelmente insuficiente para alterar o equilíbrio endêmico da doença, portanto, precisa ser reavaliado envolvendo no processo todos os agentes públicos e privados interessados no tema.(AU)

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The state of Bahia has the third largest equine herd in Brazil, with the horse industry overtaking several sectors of the economy currently. However, animals may be exposed to several pathogenic agents, especially the equine infectious anemia (EIA) virus. EIA is a persistent viral disease with a worldwide distribution and is considered the main infectious disease in horses. Therefore, this study sought data from serological survey EIA in Itapé and Barro Preto municipalities, inserted in the southern Coastal Identity Territory of Bahia, by using the agar gel immunodiffusion test. A total of 1,880 blood serum samples from Equidae older than 6 months were analyzed from 172 properties. The general prevalence of outbreaks was 25.58% (44/172), presenting 40.47% (34/84) in Barro Preto (cocoa zone) and 11.36% (10/88) in Itapé (livestock zone). The prevalence of positive animals in Itapé was 1.01% and in Barro Preto 12.15%. Each assessed property received a questionnaire with emphasis on sanitary management and a booklet containing basic disease information. It was concluded the EIA is endemic in two epidemiologically distinct areas in the Southern Coastal Identity Territory of Bahia, one with a high prevalence in the cocoa zone for both animals and properties, and the other, in livestock zone, with a significantly low prevalence for animals, however also high for properties. Regarding the risk factors, there was a correlation between the species and age of the animals. Mules and the elderly showed a significant association, probably due to the habitat they live in and longer exposure to the virus.

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Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

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Small ruminant lentiviruses (SRLV) are difficult to diagnose due to their escape mechanisms. Therefore, proteomics is an alternative in the search for biomarkers through extracellular matrix metalloproteinases (MMPs), enzymes related to the immune response. In this sense, this study aimed to analyze the profile of MMPs in healthy and infected Toggenburg goats with chronic SRLV infection in Southeast Brazil. Five positive and five negative goats for SRLV were selected using the agar gel immunodiffusion (AGID) microtechnique, western blot (WB), and nested polymerase chain reaction (nPCR). All animals were submitted to blood collection by puncture of the jugular vein, followed by centrifugation to obtain blood plasma, protein quantification by the Bradford method, one-dimensional electrophoretic separation (1D), and identification of protease activity by zymography and confirmation via reverse zymography in the presence of MMP-2 through the action of tissue inhibitors (TIMP-2). The analysis of protein bands was performed using descriptive statistics and densitometry values for zymography were subjected to the Shapiro-Wilk test to determine normality. Little difference was observed in the occurrence of protein bands between groups. Regarding MMPs, no differences were observed in the expression of proMMP-9, MMP-9, and MMP-2 in animals affected by SRLV. TIMP-2 inhibited proMMP-2 and MMP-2 in all animals. Thus, the profile of protein bands does not change in healthy goats with chronic SRLV infection. The TIMP-2 expression allowed proving the existence of MMP-2 in animals chronically infected by SRLV via reverse zymography

Lentivírus de pequenos ruminantes (LVPR) demonstram diagnóstico complexo devido seus mecanismos de escape. Desse modo, a proteômica apresenta-se como alternativa na busca por biomarcadores através das metaloproteinases da matriz extracelular (MMPs), enzimas ligadas a resposta imunológica. Assim, objetivou-se analisar o perfil das MMPs em cabras Toggenburg sadias e com infecção crônica por LVPR no Sudeste brasileiro. Selecionou-se cinco cabras positivas e cinco negativas para LVPR utilizando: microtécnica de imunodifusão em gel de agarose (MIDGA), Western Blot (WB) e reação em cadeia da polimerase nested (nPCR). Todas foram submetidas à coleta de sangue por punção da veia jugular, seguido de centrifugação para obtenção do plasma sanguíneo, quantificação proteica pelo método Bradford, separação via eletroforese unidimensional (1D), e identificação da atividade das proteases por zimografia e confirmação via zimografia reversa na presença da MMP-2 por meio da ação de inibidores teciduais (TIMP-2). A análise das bandas proteicas ocorreu através de estatística descritiva e para a zimografia os valores de densitometria foram submetidos ao teste de Shapiro-Wilk para determinar a normalidade. Observou-se pouca distinção na ocorrência das bandas proteicas entre os grupos. Em relação as MMPs, não houve diferenças na expressão da proMMP-9, MMP-9 e MMP-2 nos acometidos por LVPR. Observou-se que a TIMP-2 inibiu a proMMP-2 e MMP-2 em todos os animais. Dessa forma, o perfil de bandas proteicas não se altera em cabras sadias e com infecção crônica por LVPR. Através da expressão da TIMP-2 foi possível comprovar a existência da MMP-2 em animais cronicamente infectados por LVPR via zimografia reversa

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The objective of this work was to describe the first record of antibodies to the Bluetongue Virus (BTV) in ewe, in the state of Amazonas. The ewe, which was in twin pregnancy, gave birth on May 9, 2015, but a lamb died hours after delivery. Veterinary service was then requested by the owner, where emaciation, loss of wool, pyrexia, apathy, dyspnea, mucoid nasal secretion, facial, lingual and submandibular edema were observed. There was a visit by the Agricultural Defense Agency of the State of Amazonas to the property and blood samples were collected from the animal. The whole blood and serum were sent to the National Agricultural Laboratory, where it was possible to detect the presence of specific antibodies to BTV, through the Agar Gel Double Immunodiffusion. The ewe was submitted to a new blood collection, following the same protocols and the samples were sent to the Biological Institute of São Paulo, confirmed diagnosis. The animal in a serious clinical condition, could not resist and died in July 2015. The occurrence of an allochthonous case, in an area where vector insects occur, can trigger an endemic process in the Amazon region. With this, the epidemiological control of these occurrences is necessary, in order to avoid the spread of the disease in the country.

O objetivo do trabalho foi descrever o primeiro registro de anticorpos para o Vírus da Língua Azul (VLA) em ovino, no estado do Amazonas. A ovelha, que se encontrava em gestação gemelar, pariu no dia 9 de maio de 2015, porém um cordeiro faleceu horas após o parto. Foi então solicitado serviço veterinário por parte do proprietário, onde foi observado emaciação, perda de lã, pirexia, apatia, dispneia, secreção nasal mucoide, edema facial, lingual e submandibular. Houve visita da Agência de Defesa Agropecuária do Estado do Amazonas na propriedade e coletadas amostras de sangue do animal. O sangue total e soro foram enviados ao Laboratório Nacional Agropecuário, no qual foi possível detectar a presença de anticorpos específicos para VLA, através do teste de Imunodifusão Dupla em Gel de Ágar. A ovelha foi submetida a uma nova coleta de sangue, seguindo os mesmos protocolos e as amostras foram enviadas ao Instituto Biológico de São Paulo, confirmando diagnóstico. O animal em estado clínico grave, não resistiu e veio a óbito em julho de 2015. A ocorrência de um caso alóctone, em uma área de ocorrência de insetos vetores, pode desencadear um processo de endemia na região amazônica. Com isso, o controle epidemiológico destas ocorrências, se fazem necessários, afim de se evitar a disseminação da doença no país.

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The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Resumo

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Resumo

Brucella ovis, a non-zoonotic species, is the etiological agent of ovine brucellosis, an infectious disease of clinical or subclinical occurrence in sheep flocks. Until then, there is no serological study of anti-Brucella ovis antibodies in purebred sheep herds. This study aimed to determine the presence of anti-Brucella ovis antibodies in purebred sheep flocks with breeding purposes from Parana State. Blood samples from 728 animals, of which 563 were females and 165 males, between 8 and 56 months of age from the six major sheep producing mesoregions of Parana, were submitted to detection of anti-Brucella ovis antibodies by the Agar Gel Immunodiffusion technique using an antigen from the bacteria Brucella ovis (Reo 198). The results indicate the presence of this disease in purebred sheep from Parana State in a low occurrence of 0.27% (2/728). The only two positive animals were rams, Santa Inês breed, from the same flock in the East Center region of Parana, without clinical disease. In conclusion, Brucella ovis is present in purebred sheep in Parana State, Brazil, and this low occurrence may have occurred due to rigorous breeding systems that may contribute to reduce the transmission of this disease.(AU)

Brucella ovis, espécie não zoonótica, é o agente etiológico da brucelose ovina, doença infecciosa de ocorrência clínica ou subclínica. Atualmente, não existe estudo sorológico de anticorpos anti-Brucella ovis em rebanhos de ovinos puros de origem. Este estudo teve como objetivo determinar a presença de anticorpos anti-Brucella ovis em rebanhos ovinos de raça pura de origem, com fins reprodutivos do estado do Paraná. Amostras de sangue de 728 animais, sendo 563 fêmeas e 165 machos, entre oito e 56 meses de idade, pertencentes a seis principais mesorregiões produtoras de ovinos no Paraná, foram submetidas à detecção de anticorpos anti-Brucella ovis pela técnica de imunodifusão em ágar gel usando-se um antígeno da bactéria Brucella ovis (Reo 198). Os resultados indicam a presença da doença em ovinos puros de origem do estado do Paraná em baixa ocorrência de 0,27% (2/728). Os dois únicos animais positivos foram reprodutores da raça Santa Inês, do mesmo rebanho da região Centro Leste do Paraná, sem manifestação clínica. Em conclusão, Brucella ovis está presente em ovinos puros de origem no estado do Paraná, e essa baixa ocorrência pode ter ocorrido devido a sistemas rigorosos de criação, que podem contribuir para a redução da transmissão dessa doença.(AU)

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Mycobacterium avium subesp. paratuberculosis (MAP) e o vírus da leucemia bovina (BLV) são agentes que causam grandes perdas econômicas nos rebanhos. O objetivo deste estudo foi avaliar a situação epidemiológica da paratuberculose bovina (PTB) e leucose enzoótica bovina (EBL) em rebanhos leiteiros de Lagoa Formosa, Minas Gerais, Brasil. Foram coletadas 236 amostras de sangue de vacas, as quais foram submetidas aos testes ELISA e imunodifusão em gel de ágar para detecção de anticorpos contra MAP e BLV. A soroprevalência de anticorpos contra MAP e BVL foi de 20% para os rebanhos e 6% para os animais e de 85% para os rebanhos e 50,42% para os animais, respectivamente. A presença dessas enfermidades deve servir como um alerta para os produtores e veterinários, para que concentrem maior atenção na implementação de medidas higiênico-sanitárias, incorporando elementos de vigilância com base nos riscos identificados no estudo.(AU)

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There are several factors affecting global milk production leading to severe economic losses. Mastitis and leukosis are highly prevalent diseases especially in Brazilian cattle herds. The present study aimed to evaluate the correlation between mastitis and the bovine leukemia virus (BLV) infection in naturally infected cows belonging to dairy herds in Minas Gerais State. Additionally, the prevalence of both diseases, pathogens causing mastitis, and their association with BLV infection and persistent lymphocytosis were evaluated. The prevalence of BLV was determined using the agar gel immunodiffusion test (AGID) in 1,392 lactating cows from ten dairy herds. From the AGID results, 240 cows were selected and divided into two experimental groups: A case group composed of 160 BLV-positive cows (BLV-P) and the control group of 80 BLV-negative cows (BLV-N). Based on serological and leucocytic evaluations, the animals were classified into three subgroups: BLV-N, BLV-P without persistent lymphocytosis (BLV-PWPL), and BLV-positive with persistent lymphocytosis (BLV-PPL). The California Mastitis Test (CMT) was performed on days 0 (D0) and 60 (D60) in the subgroups and the milk from CMT-positive glands were analyzed. BLV infection was present in 100% of herds, with an overall prevalence of 80.9% in cows older than 24 months. Among BLV-P animals, 74% were classified as BLV-PWPL and 26% as BLV-PPL. Coagulase-negative Staphylococcus (73.30%), Corynebacterium spp. (12.04%), Streptococcus agalactiae (8.90%), and Staphylococcus aureus (5.76%) were predominant (61.41%). There were no statistical differences among the scores of clinical or subclinical mastitis with the pathogen type (contagious or environmental) in the BLV-PWPL or BLV-PPL subgroups. However, BLV-P animals showed higher CMT scores than BLV-N animals, suggesting a higher predisposition to subclinical mastitis in cows affected by leukosis.(AU)

Tendo em vista a relevância econômica e social da produção de leite e a ampla ocorrência da leucose e mastite em rebanhos brasileiros, foi proposto o presente estudo com o objetivo de avaliar a correlação entre a mastite e a infecção pelo vírus da leucose bovina em vacas leiteiras naturalmente infectadas de 10 rebanhos leiteiros da região sul do estado de Minas Gerais. Além disso, foi avaliada a prevalência de ambas as doenças e dos patógenos causadores de mastite e sua associação com infecção por BLV e linfocitose persistente. A prevalência do VLB foi avaliada pelo teste de imunodifusão em gel de ágar (IDGA) em 1.392 vacas em lactação nos rebanhos estudados. De acordo com os resultados do IDGA, 240 vacas foram selecionadas para compor dois grupos experimentais: 1-caso: composto por 160 vacas BLV positivas (BLV-P) e 2-controle: composto por 80 vacas BLV negativas (BLV-N). De acordo com as avaliações sorológicas e leucocitárias, os animais foram classificados em três subgrupos: BLV-N; BLV-P sem linfocitose persistente (BLV-PWPL) e BLV positivos com linfocitose persistente (BLV-PPL). O California Mastitis Test (CMT) e a cultura do leite das glândulas CMT positivas foram realizados nos dias 0 (D0) e 60 (D60). A leucose foi detectada em 100% dos rebanhos com prevalência média de 80,9%. Entre os animais BLV-P, 74% foram classificados como BLV-PWPL e 26% como BLV-PPL. Staphylococcus coagulase negativos (73,30%), seguido por Corynebacterium spp. (12,04%), Streptococcus agalactiae (8,90%) e Staphylococcus aureus (5,76%) foram os agentes predominantes. Não houve diferença estatística entre os escores de mastite clínica ou subclínica e nem entre o tipo de patógeno (contagioso ou ambiental) nos subgrupos de animais BLV-PWPL ou BLV-PPL relacionados ao BLV-N. No entanto, animais BLV-P apresentaram escores mais elevados no CMT em relação aos animais BLV-N, sugerindo maior predisposição à mastite subclínica em vacas com leucose.(AU)

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Background: Small ruminant lentivirus (SRLV) belong to genus Lentivirus, family Retroviridae. These viruses causecaprine arthritis encephalitis (CAE) and maedi visna (MV), infectious diseases that cause economic, production, and reproductive losses. There are no effective treatments or vaccines for these diseases. Thus, early detection via serology hasgreat importance for control of SRLV. Therefore, the objective of this review is to demonstrate the potential of the westernblot (WB) test as an immunodiagnostic test for SRLV.Review: In general, immunodiagnosis of SRLV is performed via agar gel immunodiffusion (AGID) and indirect enzymelinked immunosorbent assay (ELISA), which can detect antibodies in several different biological samples but is used preferably with serum and blood plasma. However, WB has demonstrated efficacy in the early diagnosis of immunoglobulinsagainst SRLV, presenting higher sensitivity and specificity than the serological tests usually used, because this techniquecan detect antibodies at a dilution as much as 256 times greater than that of AGID and 32 times greater than that of ELISA.SRLV infection and consequent immunological activation result in the induction of cellular and humoral responses. Additionally, around the third week, production of antibodies directed mainly toward viral capsid proteins (p25 and p28)occurs. After the fifth week, production of immunoglobulins directed toward other viral proteins occurs. Because of thepersistence of SRLV infection, serology is considered to be the most practical means to diagnosis. Each serological testhas a percentage specificity and distinct sensitivity, as well as advantages and disadvantages in its applicability. It shouldbe noted that there is no gold standard test for diagnosis of SRLV infection. Moreover, SRLV are characterized by escapemechanisms such as genetic diversity, mutagenic potential, viral intermittence...

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Equine infectious anemia (EIA) is a viral infection, caused by a lentivirus of the Retroviridae family, Orthoretrovirinael subfamily and its occurrence generates significant economic losses due to culling of positive animals as a measure of infection control. The objective of this work was to determine the prevalence of horses positive for equine infectious anemia virus (EIAV) and to identify the occurrence of areas with higher densities of cases in the states of Paraíba (PB), Pernambuco (PE), Rio Grande do Norte (RN) and Ceará (CE), Northeast region of Brazil, during the rainy (May and June) and dry (October and November) periods of 2017 and 2018. Serum samples from 6,566 horses from the states of PB, PE, RN and CE, Brazil, provided by the Laboratório Veterinária Diagnóstico - Ltda., were used. Serological diagnosis of EIA was performed using indirect enzyme-linked immunosorbent assay (ELISA) as a screening test and agar gel immunodiffusion test (AGID) as a confirmatory test. The apparent prevalence was obtained by dividing the number of seroreactive animals by the total number of animals, while the true prevalence was estimated by adjusting the apparent prevalence, considering the sensitivity (100%) and specificity (98.6%) of the diagnostic protocol used. For the construction of Kernel estimates, the Quartic function was used. In the dry season, of the 1,564 animals sampled, 28 were serologically positive, of which 19 belonged to the state of Ceará, 7 to Paraíba and 2 to Rio Grande do Norte. In 2018, it was observed that, during the rainy season, 26 of the 1,635 horses were seroreactive, with 19 cases resulting from Ceará, 4 from Paraíba and 3 from Pernambuco. In the dry season, 32 of the 1,526 animals were seroreactive to EIAV, of which 26 were from Ceará, 3 from Paraíba, 1 from Rio Grande do Norte and 2 from Pernambuco. In the dry period of 2017, the CE had a real prevalence of 1.22% (95% CI = 0.05 - 2.99%). In 2018, during the rainy season, prevalences of 0.03% (95% CI = 0 - 1.18%) were identified in CE and 1.69% (95% CI = 0 - 8.38%) in PE. Regarding the 2018 dry period, a prevalence of 1.32% (95% CI = 0.26 - 2.84%) was found in the state of CE. In both dry and rainy periods of 2017, the presence of spatial clusters of animals positive for EIA was observed, mainly in the border areas among the states of CE, PE, PB and RN. In 2018, there was a variation in the distribution of areas with higher densities of cases between the rainy and dry periods. The state of CE had the highest prevalence of positive animals and the presence of areas with higher densities of EIA cases in both climatic periods, in the years 2017 and 2018. In some municipalities of the CE, important sporting events of agglomeration of animals take place, which can favor the transmission of EIAV by facilitating the contact of infected and susceptible animals. Population density may be a factor associated with the higher prevalence observed in this region, as it has the second largest herd among the states studied. Higher densities indirectly contribute to the occurrence of infectious diseases, as they favor the contact of infected and susceptible animals. The occurrence of higher densities of cases in the border areas of the states of PE, RN, CE, and PB may be related to the greater movement of animals in these regions, favoring the indirect contact of infected horses with susceptible ones. The observed results demonstrate the circulation of the EIAV in four states in the Northeast region of Brazil.(AU)

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Background: The enzootic bovine leukosis (EBL) is a viral disease with a chronic aspect and its etiological agent is anoncogenic deltaretrovirus called bovine leukemia virus (VLB). It has a wide distribution in dairy cattle herds, and it maytake several years before the first clinical manifestations occur. Animals seropositive for VLB are more susceptible todeveloping infectious diseases such as mastitis. The objective of this study was to evaluate the seroprevalence of EBL, theprevalence of mastitis, the possible association between leukosis and mastitis, and the risk factors related to leukosis inmunicipalities of the Caparaó region of Espírito Santo, Brazil.Materials, Methods & Results: The prevalence of enzootic bovine leukosis and its association with mastitis were evaluatedin an 854 clinically healthy crossbred (Bos tauros tauros x Bos taurus indicus) dairy cows were used, in different lactationphases, from farms located in the 12 municipalities that compose the Caparaó Capixaba micro-region, in the southernpart of Espírito Santo, Brazil, in the period from February to December 2015. The seroprevalence of EBL was determinedby the agar gel immunodiffusion test, the presence of clinical mastitis was ascertained by the black background mug testand of subclinical mastitis by the CMT. The reuse of needles, type of milking, reproductive management, calf mortality,presence of reproductive problems and milk yield were the risk factors assessed. The association between the variableswas estimated by the odds ratio (95% confidence interval). The significance of the associations was determined by theChi-square test. The prevalence of EBL, clinical and subclinical mastitis was 56.79%, 5.50% and 43.55%, respectively.There was a positive association between the EBL virus and the presence of...

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Background: Bluetongue is a vector-borne viral disease transmitted by midges from the genus Culicoides. The disease caninfect most of the ruminant and camelid species, but the severe disease is most often seen in european wool and muttonsheep breeds. In this sense, there is a gap in the knowledge on BTV infection in hair sheep breeds from tropical zones.Thus, this study aimed at establishing whether exposure to BTV is a risk factor for reproductive failure in Santa Inês ewes,a hair sheep breed, reared under tropical conditions in Brazil.Materials, Methods & Results: A retrospective cross-sectional study was carried out in sheep farms in São Paulo state,Brazil, after the rainy season. Serum samples from 110 Santa Inês ewes with a history of reproductive disorders, in the last6 months, which were included: abortion, premature birth, stillbirth, retention of placenta, infertility, estrus repetition, fetalmalformation, weak lamb birth and neonatal death were collected. The presence of antibodies against BTV was assessedby agar gel immunodiffusion method (AGID). Serology to the infectious agents Brucela ovis, Lepstopira spp., Toxoplasmagondii, Neospora caninum and Campylobacter sp. were also assessed. Bivariate associations between the outcome andindividual explanatory variables were assessed using the Fisher’s exact test. Abortion was the most common reproductive disorder (53%; 74/139) observed, followed by estrus repetition (12%; 17/139) and infertility (11%; 15/139). Otherdisorders related to the conceptus totaled nearly one fourth of the reported disorders. A total of 20% (22/110) of the eweswere seropositive to BTV. A higher frequency of BTV seropositive than BTV seronegative ewes with a history of abortionwas found. Also, abortion with seroreactivity to BTV was tested for prevalence ratio that showed 1.38 [95% CI 1.10-1.74;P = 0.030]. With regards to the abortion involvement of other infectious diseases associated with the seropositive ewes to...

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Equine infectious anemia (EIA) is one of the most important diseases from the health and economic point of view for equidae breeding, as it does not have treatment and vaccines. The Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA) instituted mandatory sanitary measures that include the official diagnosis by the agar gel immunodiffusion (AGID) test and sacrifice of seropositive animals to control this disease. Seventy-two seronegative equines, challenged with different vaccines, were used to verify the occurrence of non-specific reactions in the AGID techniques. Five serological controls were performed one week after vaccination, at seven-day intervals. The results indicated that the use of vaccines in equines in a period that precedes the performance of laboratory tests for the diagnosis of EIA does not induce seroconversion. However, 11.11% of the equines vaccinated against influenza, encephalomyelitis, equine rhinopneumonitis, and tetanus, and 15.38% of those vaccinated against leptospirosis had non-specific negative reactions to AGID. In this study, there was a non-specific line in the AGID for EIA described by Ordinance No. 84/1992 by MAPA but already mentioned in the Normative Instruction 55 of 26 November 2018.

Anemia Infecciosa Equina é uma das enfermidades mais importantes sob o ponto de vista sanitário e econômico para a equideocultura, por não possuir tratamento e vacinas. Para controle desta doença, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) instituiu medidas sanitárias obrigatórias em todo território nacional que incluem o diagnóstico e sacrifício dos animais soropositivos. Para verificar a ocorrência de reações inespecíficas na técnica de IDGA utilizou-se 72 equinos soronegativos, desafiados com diferentes vacinas. Uma semana após a vacinação, realizou-se cinco controles sorológicos, em intervalos de sete dias. Os resultados indicaram que o uso de vacinas em equinos em período que antecede a realização de exames laboratoriais para diagnóstico de AIE, não induz a soroconversão. Entretanto, 11,11% dos equinos vacinados contra influenza, encefalomielite, rinopneumonite equina e tétano, e 15,38% dos que foram vacinados contra leptospirose apresentaram reações negativas inespecíficas ao IDGA. Neste estudo, verificou-se uma linha inespecífica no IDGA para AIE.

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The present study aimed to estimate the prevalence of antibodies against Brucella ovis-epididymitis, smooth-Brucella, leptospirosis, toxoplasmosis and Maedi-visna in sheep slaughtered in Minas Gerais, Brazil and to study their simultaneous occurrence, including caseous lymphadenitis, at sheep and flock levels. The study was conducted at a sheep slaughterhouse with Federal Inspection Service. Sera from 594 animals from 21 flocks were collected, in 2007. The agar gel immunodiffusion (AGID) was employed to detect anti-B. ovis and anti-Maedi Visna antibodies, whereas Rose Bengal (RB) and the 2-mercaptoethanol test (2ME) were used to test anti-smooth Brucella antibodies. For the detection of anti-Leptospira antibodies, sera were examined by microscopic agglutination test (MAT), while for the detection of IgG antibodies to Toxoplasma gondii ELISA was used. Prevalence of antibodies against smooth Brucella, B. ovis-epididimitis, Leptospira spp., toxoplasmosis and Maedi-Visna found in sheep from Minas Gerais was 0.00%, 24.04%, 25.96%, 10.46% and 3.08%, respectively; whereas the seroprevalence in flocks was 0.00%, 80.95%, 90.48%, 71.43% and 23.81%, respectively. Moreover, when data on antibodies anti-Corynebacterium pseudotuberculosis, previously obtained, were included, about 60% of the flocks showed animals that were exposed to four or more of the studied agents. However, only 25.47% o

O presente estudo teve como objetivo estimar a prevalência de anticorpos contra Brucella ovis (epididimite ovina), Brucella lisa, leptospirose, toxoplasmose e Maedi-visna em ovinos abatidos em Minas Gerais, Brasil, e estudar sua ocorrência simultânea, incluindo linfadenite caseosa, nos ovinos e nos rebanhos. O estudo foi realizado em um abatedouro de ovinos com Serviço de Inspeção Federal. Soros de 594 animais de 21 rebanhos foram coletados, em 2007. A imunodifusão em gel de ágar (IDGA) foi empregada para detectar anticorpos anti-B. ovis e anticorpos anti-Maedi Visna, enquanto o teste do antígeno acidificado tamponado (AAT) e o teste de 2-mercaptoetanol (2ME) foram utilizados para testar anticorpos anti-Brucella lisa. Para a detecção de anticorpos anti-Leptospira, os soros foram examinados pelo teste de aglutinação microscópica (MAT), enquanto que para a detecção de anticorpos IgG para Toxoplasma gondii, foi usado o ELISA. A prevalência de anticorpos anti-Brucella lisa, B. ovis, Leptospira spp., toxoplasmose e Maedi-Visna encontrados em ovinos de Minas Gerais foi de 0,00%, 24,04%, 25,96%, 10,46% e 3,08%, respectivamente; enquanto a soroprevalência em rebanhos foi de 0,00%, 80,95%, 90,48%, 71,43% e 23,81%, respectivamente. Além disso, quando dados de anticorpos anti-Corynebacterium pseudotuberculosis, previamente obtidos, foram incluídos, cerca de 60% dos rebanhos apresentaram a

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Caprine arthritis encephalitis and Maedi-Visna are lentiviruses affecting goats and sheep, respectively. Despite the literature having studies about these diseases, there is a constant demand and the need to study the health status of flocks that exploit economically. Therefore, this study aimed to assess the frequency of small ruminant lentiviruses explored in regional locations of Chapadinha and Itapecuru Mirim, that compose the microregion of Low Parnaíba, Maranhão, Brazil, as well as analyze the spatial distribution of outbreaks in the studied regions. Therefore, 241 properties were visited, where blood was collected in 1150 sheep and 1260 goats and tested by agar gel immunodiffusion (AGID). Epidemiological questionnaire was applied and collected the geographic coordinates. There was a low frequency for lentivirus, with 0.39% (5/1260) of goats and 0.08% (1/1150) of sheep. Regarding the spatial analysis, the reagent flocks were distributed in strategic cities for commercialization throughout the microregion. There was a low occurrence of lentiviruses. The municipalities of Cantanhede and Pirapemas of the regional of Itapecuru Mirim and Brejo and Magalhães de Almeida had reagent flocks for CAE. Whereas the municipality of Matões do Norte presented flock reagent to Maedi-Visna, this belonging to the regional of Chapadinha.

A artrite encefalite caprina e a Maedi-Visna são lentivírus que afetam cabras e ovelhas, respectivamente. Apesar da literatura apresentar estudos sobre essas doenças, existe uma demanda constante e a necessidade de se estudar o estado de saúde dos rebanhosexploradoseconomicamente. Portanto, este estudo teve como objetivo avaliar a frequência de lentivírus de pequenos ruminantes explorados nasregionais de Chapadinha e Itapecuru Mirim, que compõem a microrregião do Baixo Parnaíba, Maranhão, Brasil, bem como analisar a distribuição espacial dos surtos nas regiões estudadas. Para tanto, foram visitadas 241 propriedades, onde foi coletado sangue de 1150 ovelhas e 1260 cabras e testado por imunodifusão em gel de ágar (IDGA). Foi aplicado questionário epidemiológico e coletado as coordenadas geográficas. Houve baixa frequência para lentivírus, com 0,39% (5/1260) de caprinos e 0,08% (1/1150) de ovinos. Quanto à análise espacial, os rebanhos reagentes foram distribuídos em cidades estratégicas para comercialização em toda a microrregião. Houve baixa ocorrência de lentivírus. Os municípios de Catanhede e Pirapemas da regional de Itapecuru Mirime Brejo e Magalhães de Almeida possuíam rebanhos reagentes para CAE. Já o município de Matões do Norte apresentou reagente rebanho à Maedi-Visna, esta pertencente à regional de Chapadinha.

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Among the diseases that affect the reproductive system of domestic animals, brucellosis in the sheep species is important because it generates significant economic losses to sheep rearing. Thus, it is a threat to the growth and productivity of sheep herds. In the face of this problem, the objective of the present research was to identify the prevalence of ovine brucellosis in herds in municipalities of the Teresina, Piauí, Brazil microregion by using the agar gel immunodiffusion assay (AGID) and indirect enzyme-linked immunosorbent assay (ELISA) serological tests. Fourteen municipalities were included in the research. Blood samples were collected from 521 pubescent animals by puncturing the jugular vein. After collection, the samples were submitted to the serological techniques, AGID and indirect ELISA, to detect anti B. ovis antibody. Of the 521 samples submitted to the AGIDtest, 23 (4.41%) were sera reagent and 498 (95.58%) negative. The indirect ELISA tests, 24 (4.61%) suspect samples and 497 (95.39%) negative samples were obtained, and there were no reagent animals in this test, only suspect. The seroprevalence of ovine brucellosis in the Teresina, Piauí microregion was 4.41%. Thus, it is possible to identify sheep with reagent serology to infection by B. ovis, that is present in municipalities in the state of Piauí, Brazil. Furthermore, AGIDwas shown to be more sensitive in detecting animals that had had contact with the etiological agent of the disease.(AU)

Dentre as enfermidades que acometem o sistema reprodutivo dos animais domésticos, a brucelose na espécie ovina tem se destacado por gerar prejuízos econômicos significativos à ovinocultura. Dessa forma, apresenta-se como uma ameaça ao crescimento e à produtividade dos rebanhos ovinos. Diante de tal problemática, objetivou-se, por meio desta pesquisa, identificar a prevalência de brucelose ovina em rebanhos pertencentes a municípios da microrregião de Teresina, Piauí, por meio dos testes sorológicos, imunodifusão em gel de ágar (IDGA) e ensaio imunoenzimático (ELISA) indireto. Quatorze municípios foram incluídos na pesquisa. Para sua execução, colheram-se, por punção venosa da jugular, amostras sanguíneas de 521 animais púberes. Após colheita, as amostras foram submetidas às técnicas sorológicas, IDGA e ELISA indireto, para a detecção de anticorpos anti-B. ovis. Das 521 amostras submetidas ao teste de IDGA, 23 (4,41%) foram sororreagentes e 498 (95,58%) negativas. Quanto ao teste ELISA indireto, obtiveram-se 24 (4,61%) amostras suspeitas e 497 (95,39%) amostras negativas, não havendo animais reagentes neste teste, apenas suspeitos. A soroprevalência da brucelose ovina na microrregião homogênea de Teresina, Piauí, foi de 4,41%. Assim, foi possível identificar ovinos com sorologia reagente à infecção por B. ovis, presente em municípios do estado do Piauí. Além disso, a IDGA mostrou-se mais sensível em detectar animais que tiveram contato com o agente etiológico da doença.(AU)

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Bluetongue is an infectious, non-contagious disease that affects domestic and wild ruminants, caused by a virus from the Orbivirus genus, Reoviridae family, transmitted by arthropod vectors of the Culicoides genus. This paper aims to be the first serological survey of bluetongue in sheep from the Meso-regions of Campo das Vertentes and South and Southeast of Minas Gerais. Samples were collected from sheep from different properties. The serum samples were submitted to Agar Gel Immunodiffusion (AGID) and competitive Enzyme-Linked Immunosorbent Assay (cELISA). 303 serum samples were submitted to AGID and cELISA. In these samples, 164 (54.13%) were positive in the AGID technique, and 171 (56.44%) positive in the cELISA technique, with an almost perfect agreement between the techniques (kappa index = 0.887). In all visited properties, positive animals have been found in the herd. Animals acquired from properties of the studied mesoregions were more likely to be positive in IDGA and cELISA tests than animals acquired from properties in other regions of Brazil (p<0.001). These results suggest that bluetongue virus (BTV) is widespread in the mesoregions of Campo das Vertentes and South and Southeast of Minas Gerais.(AU)

A língua azul (LA) é uma doença infecciosa, não contagiosa, que acomete ruminantes domésticos e silvestres, causada por um vírus do gênero Orbivirus da família Reoviridae, transmitida por vetores artrópodes do gênero Culicoides. O presente estudo representa o primeiro trabalho a realizar um inquérito sorológico da língua azul em rebanhos ovinos nas Mesorregiões de Campo das Vertentes e Sul e Sudoeste de Minas Gerais. Foram coletadas amostras de soro de ovinos de diferentes propriedades. As amostras de soro foram submetidas aos testes de imunodifusão em gel de ágar (IDGA) e ensaio de imunoadsorção enzimática por competição (cELISA). Ao todo 303 amostras de soro foram submetidas ao IDGA e cELISA. Dessas amostras, 164 (54,13%) foram positivas na técnica de IDGA e 171 (56,44%) positivas na técnica de cELISA, havendo concordância quase perfeita entre as técnicas (índice kappa = 0,887). Em todas as propriedades visitadas, foram encontrados animais positivos no rebanho. Animais adquiridos de propriedades das Mesorregiões estudadas, tiveram mais chances de serem positivos nos testes de IDGA e cELISA do que animais adquiridos de propriedades de outras Regiões do Brasil (p<0,001). Esses resultados sugerem que o vírus da língua azul encontra-se disseminado em ovinos nas Mesorregiões de Campo das Vertentes e Sul e Sudoeste de Minas Gerais.(AU)