RESUMO

This study aimed to evaluate, in vitro, the use of leaf extracts of Azadirachta indica (A. indica) and Melia azedarach (M. azedarach) as antivirals against caprine lentivirus (CLV) in colostrum and milk of goat nannies. These were collected from eight individuals and infected with the standard strain of CLV. Samples were then subdivided into aliquots and treated with 150 µg/mL of crude extract, and with ethyl acetate and methanol fractions for 30, 60, and 90 min. Next, somatic cells from colostrum and milk were co-cultured with cells from the ovine third eyelid. After this step, viral titers of the supernatants collected from treatments with greater efficacy in co-culture were assessed. The organic ethyl acetate fractions of both plants at 90 min possibly inhibited the viral activity of CLV by up to a thousandfold in colostrum. In milk, this inhibition was up to 800 times for the respective Meliaceae. In conclusion, the ethanolic fraction of ethyl acetate from both plants demonstrated efficacy against CLV in samples from colostrum and milk when subjected to treatment, which was more effective in colostrum.

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This study aimed to evaluate by means of Nested Polymerase Chain Reaction (nPCR), co-cultivation and sequencing, with genetic comparison between strains (mother/newborn), the occurrence of vertical transmission of Small Ruminant Lentiviruses (SRLV) from naturally occurring nannies infected for their offspring. For the detection of SRLV seropositive progenitors, blood was collected from 42 nannies in the final third of gestation in tubes with and without anticoagulant. The diagnostic tests used were Western Blot (WB) and nPCR. During the period of birth, the same blood collection procedure was performed on 73 newborns at zero hours of birth, with the same diagnostic tests. Seventeen blood samples from seven-day-old kids, proven positive for SRLV by nPCR, chosen at random, were subjected to coculture in goat synovial membrane (GSM) cells for 105 days. The pro-viral DNA extracted from the cell supernatant from the coculture was subjected to nPCR. For DNA sequencing from the nPCR products, nine positive samples were chosen at random, four nannies with their respective offspring, also positive. Each sample was performed in triplicate, thus generating 27 nPCR products of which only 19 were suitable for analysis. Among the 42 pregnant goats, in 50% (21/42) pro-viral DNA was detected by nPCR, while in the WB, only 7.14% (3/42) presented antibodies against SRLV. Regarding neonates, of the 73 kids, 34 (46.57%) were positive for the virus, using the nPCR technique, while in the serological test (WB), three positive animals (4.10%) were observed. The coculture of the 17 samples with a positive result in the nPCR was confirmed in viral isolation by amplification of the SRLV pro-viral DNA. When aligned, the pro-viral DNA sequences (nannies and their respective offspring) presented homology in relation to the standard strain CAEV Co. It was concluded that the transmission of SRLV through intrauterine route was potentially the source of infection in the newborn goats.

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Background: Caprine Arthritis Encephalitis (CAE) is a disease that causes productive losses in dairy goat flocks due to thereduction in milk production, followed by lesions in joints and mammary glands. An early diagnosis is essential, consideringthat there is frequent occurrence of asymptomatic animals. Hence, this study aimed to perform a comparison of immunological and molecular based diagnostic tests, represented by Agar Gel Immunodiffusion (AGID), Western Blot (WB) andnested Polymerase Chain Reaction (nPCR). In addition, the mammary glands (MG) of dairy goats were clinically evaluated.Materials, Methods & Results: Blood collection and clinical examination were performed in 1191 dairy goats of 12 farms locatedin Northeastern and Southeastern regions of Brazil. Serological (AGID, WB) and molecular (nPCR) test results were comparedand the data, along with MG alterations, were analyzed using Epi-info 7 and WinEpiscope 2.0. Seroprevalence in AGID test was41.14% (490/1191). In WB, 51.47% (613/1191) of animals were seropositive and nPCR detected 69.44% (827/1191) positiveanimals. Hence, WB was more sensitive (P < 0.001) than AGID. However, nPCR detected more positive animals than AGID (P< 0.001) and WB (P < 0.001). The analysis of mammary glands revealed that 105 out of 1096 nanny goats presented alterations,of which 101 presented altered consistency, 16 presented elevated temperatures and 60 had enlarged retromammary lymph nodes.There was significant statistic difference (P < 0.05) only when comparing the results of serological tests with MG alterations.Discussion: In general, AGID technique is most frequently used when screening flocks for the disease due to the practicalityand low cost this test presents. However, the results demonstrated that AGID detected the lowest number of positive animals.This low sensitivity that the test presented may be attributed to its antigen-antibody interaction mechanism...(AU)

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RESUMO

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

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

Assuntos

RESUMO

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

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

Assuntos

RESUMO

Background: Caprine Arthritis Encephalitis (CAE) is a disease that causes productive losses in dairy goat flocks due to thereduction in milk production, followed by lesions in joints and mammary glands. An early diagnosis is essential, consideringthat there is frequent occurrence of asymptomatic animals. Hence, this study aimed to perform a comparison of immunological and molecular based diagnostic tests, represented by Agar Gel Immunodiffusion (AGID), Western Blot (WB) andnested Polymerase Chain Reaction (nPCR). In addition, the mammary glands (MG) of dairy goats were clinically evaluated.Materials, Methods & Results: Blood collection and clinical examination were performed in 1191 dairy goats of 12 farms locatedin Northeastern and Southeastern regions of Brazil. Serological (AGID, WB) and molecular (nPCR) test results were comparedand the data, along with MG alterations, were analyzed using Epi-info 7 and WinEpiscope 2.0. Seroprevalence in AGID test was41.14% (490/1191). In WB, 51.47% (613/1191) of animals were seropositive and nPCR detected 69.44% (827/1191) positiveanimals. Hence, WB was more sensitive (P < 0.001) than AGID. However, nPCR detected more positive animals than AGID (P< 0.001) and WB (P < 0.001). The analysis of mammary glands revealed that 105 out of 1096 nanny goats presented alterations,of which 101 presented altered consistency, 16 presented elevated temperatures and 60 had enlarged retromammary lymph nodes.There was significant statistic difference (P < 0.05) only when comparing the results of serological tests with MG alterations.Discussion: In general, AGID technique is most frequently used when screening flocks for the disease due to the practicalityand low cost this test presents. However, the results demonstrated that AGID detected the lowest number of positive animals.This low sensitivity that the test presented may be attributed to its antigen-antibody interaction mechanism...

Assuntos

RESUMO

The production performance of a livestock herd can be compromised by various diseases. In sheep, maedi-visna (MV) infections, which have a chronic nature, are caused by a virus (maedi-visna virus (MVV)) belonging to the genus Lentivirus of the Retroviridae family. The infection can cause significant economic losses and has considerable health impacts on sheep breeding in production systems. Due to the importance of this disease in sheep flocks, the objective was to conduct a serosurvey of MVV in the states of Ceará (CE), Rio Grande do Norte (RN), Paraíba (PB), and Sergipe (SE). A total of, 3332 serum samples were collected in the four states, 1011 in CE, 931 in RN, 459 in PB, and 931 in SE, with the number of samples proportional to the actual herd size of each state. The samples were analyzed using the agar gel microimmunodiffusion test (AGID). Reproducers were revaluated using western blotting (WB). In addition to this serological survey, we administered an investigative questionnaire to identify possible risk factors that facilitate the introduction and spread of diseases (location, category, sex, breed type, creation system, production, herd size, and association with goats). After analysis of the sera using the AGID test, there was zero prevalence. Revaluating breeders by WB revealed a 5.5% prevalence of MV in the four states studied, with prevalences for the states of CE, RN, Paraiba, and SE of 2.3% (2/88), 10.4% (8/77), 3.6% (1/28), and 4.7% (2/42), respectively, corresponding to 13 breeders containing antibodies to the virus. These findings emphasized that the choice of diagnostic tests is extremely important for the early detection of seropositive animals and thus the prevention of the spread of the virus among herds in the region.(AU)

O desempenho produtivo de um rebanho pode ser comprometido por diversas enfermidades. Em relação aos ovinos, à ocorrência da Maedi - Visna (MV) doença infecciosa, de caráter crônico, causada por um vírus (Maedi-Visna Vírus MVV) pertencente ao gênero Lentivirus da família Retroviridae. A infecção pode causar importantes perdas econômicas e elevados impactos sanitários nos sistemas de produção da ovinocultura. Em virtude da importância desta enfermidade nos rebanhos ovinos, o objetivo do estudo foi realizar um levantamento soroepidemiológico do vírus da Maedi - Visna (MVV) nos estados do Ceará (CE), Rio Grande do Norte (RN), Paraíba (PB) e Sergipe (SE). Para tanto, foram coletadas 3332 amostras de sangue nos quatro estados, sendo 1011 do CE, 931 do RN, 459 da PB e 931 de SE, sendo este número de amostras proporcional ao rebanho efetivo de cada estado. As amostras foram analisadas utilizando o teste de microimunodifusão em gel de ágar (IDGA). Os reprodutores foram reavaliados pela técnica de Western Blot (WB). Associado a esse levantamento sorológico foi aplicado um questionário epidemiológico para identificar possíveis fatores de risco que podem facilitar a introdução e disseminação de enfermidades (localização, categoria, sexo, tipo racial, sistema de criação, produção, tamanho de rebanho e associação com caprinos). Após análise dos soros pelo teste de IDGA foi verificada uma soroprevalência nula nos rebanhos estudados. Os reprodutores reavaliados pelo WB apresentaram uma prevalência geral de 5,5% da MV nos quatro estados estudados, sendo que os estados do Ceará, Rio Grande do Norte, Paraíba e Sergipe apresentaram, 2,3% (2/88), 10,4% (8/77), 3,6% (1/28) e 4,7% (2/42) respectivamente, correspondendo a 13 reprodutores que apresentaram presença de anticorpos. Diante desses resultados, ressalta-se que a escolha dos testes diagnósticos é de extrema importância para que haja uma detecção precoce de animais soropositivos, e assim evitar a...(AU)

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RESUMO

The objective of this study was to demonstrate iatrogenic transmission of small ruminant lentivirus (SRLV) from goats to sheep and horizontal transmission between sheep. The study was conducted on a farm with separate goat and sheep rearing, and animals were monitored for lentivirus occurrence by clinical examination and testing by immunoblotting (IB), agar gel immunodiffusion (AGID), and nested polymerase chain reaction (nPCR). Positive results had not been observed in the sheep flock until this study. Conversely, virus positive dairy goats were known. For this reason, the farm performed the caprine arthritis-encephalitis (CAE) control program. The study was designed with a sheep group that presented positive animals for SRLV by nPCR. It was verified that three newborn animals in this group were rejected by their mothers and consequently received milk from the goat herd. These three animals remained with another 20 sheep of the same age, totaling 23 animals. After one year, during monitoring, 11 of the 23 animals in the group presented positive results in the nPCR and three demonstrated seroconversion by IB. Of the animals that had received goat milk, two had positive results in the nPCR and IB. The 11 animals positive in the nPCR were followed and it was verified that five animals did not present further positive results in the nPCR, nor seroconversion; two continued presenting positive results in the nPCR but were negative in the IB and AGID and four were positive in the nPCR, IB, and AGID. Thus, it was possible to demonstrate iatrogenic interspecific infection and the occurrence of horizontal caprine lentivirus transmission among sheep.(AU)

O objetivo foi demonstrar a transmissão iatrogênica de lentivírus de pequenos ruminantes (LVPR) de caprinos para ovinos e a transmissão horizontal entre ovinos. O estudo foi conduzido em propriedade com criação não consorciada de caprinos e ovinos, monitorada para ocorrência de lentiviroses, a partir de acompanhamento clínico e testes de immunoblotting (IB), imunodifusão em gel de agarose (IDGA) e reação em cadeia da polimerase do tipo nested (nPCR). Resultados positivos no rebanho ovino não haviam sido observados até então. Por outro lado, sabia-se que o rebanho caprino leiteiro era positivo. Por este motivo, a propriedade realizava programa de controle da artrite-encefalite caprina (CAE). O estudo foi delineado em um lote de ovinos que apresentou animais positivos para LVPR na nPCR. Verificou-se que três animais neonatos deste lote foram rejeitados por suas mães e por isso receberam leite do rebanho caprino. Estes três animais permaneceram com outros 20 ovinos de mesma faixa etária, totalizando 23 animais. Após um ano, durante a realização do monitoramento, dos 23 animais do lote, 11 apresentaram resultados positivos na nPCR e três demonstraram soroconversão por IB. Dos animais que haviam recebido leite do rebanho caprino, dois tiveram resultados positivos na nPCR e no IB. Os 11 animais positivos na nPCR foram acompanhados, verificando-se que cinco animais não apresentaram mais resultados positivos na nPCR, nem soroconverteram; dois continuaram apresentando resultados positivos na nPCR, mas negativos no IB e IDGA e quatro apresentaram-se positivos na nPCR, no IB e no IDGA. Dessa forma, foi possível demonstrar a infecção interespecífica de forma iatrogênica e a ocorrência de transmissão horizontal do lentivírus caprino entre ovinos.(AU)

Assuntos

RESUMO

The objective of this study was to demonstrate iatrogenic transmission of small ruminant lentivirus (SRLV) from goats to sheep and horizontal transmission between sheep. The study was conducted on a farm with separate goat and sheep rearing, and animals were monitored for lentivirus occurrence by clinical examination and testing by immunoblotting (IB), agar gel immunodiffusion (AGID), and nested polymerase chain reaction (nPCR). Positive results had not been observed in the sheep flock until this study. Conversely, virus positive dairy goats were known. For this reason, the farm performed the caprine arthritis-encephalitis (CAE) control program. The study was designed with a sheep group that presented positive animals for SRLV by nPCR. It was verified that three newborn animals in this group were rejected by their mothers and consequently received milk from the goat herd. These three animals remained with another 20 sheep of the same age, totaling 23 animals. After one year, during monitoring, 11 of the 23 animals in the group presented positive results in the nPCR and three demonstrated seroconversion by IB. Of the animals that had received goat milk, two had positive results in the nPCR and IB. The 11 animals positive in the nPCR were followed and it was verified that five animals did not present further positive results in the nPCR, nor seroconversion; two continued presenting positive results in the nPCR but were negative in the IB and AGID and four were positive in the nPCR, IB, and AGID. Thus, it was possible to demonstrate iatrogenic interspecific infection and the occurrence of horizontal caprine lentivirus transmission among sheep.

O objetivo foi demonstrar a transmissão iatrogênica de lentivírus de pequenos ruminantes (LVPR) de caprinos para ovinos e a transmissão horizontal entre ovinos. O estudo foi conduzido em propriedade com criação não consorciada de caprinos e ovinos, monitorada para ocorrência de lentiviroses, a partir de acompanhamento clínico e testes de immunoblotting (IB), imunodifusão em gel de agarose (IDGA) e reação em cadeia da polimerase do tipo nested (nPCR). Resultados positivos no rebanho ovino não haviam sido observados até então. Por outro lado, sabia-se que o rebanho caprino leiteiro era positivo. Por este motivo, a propriedade realizava programa de controle da artrite-encefalite caprina (CAE). O estudo foi delineado em um lote de ovinos que apresentou animais positivos para LVPR na nPCR. Verificou-se que três animais neonatos deste lote foram rejeitados por suas mães e por isso receberam leite do rebanho caprino. Estes três animais permaneceram com outros 20 ovinos de mesma faixa etária, totalizando 23 animais. Após um ano, durante a realização do monitoramento, dos 23 animais do lote, 11 apresentaram resultados positivos na nPCR e três demonstraram soroconversão por IB. Dos animais que haviam recebido leite do rebanho caprino, dois tiveram resultados positivos na nPCR e no IB. Os 11 animais positivos na nPCR foram acompanhados, verificando-se que cinco animais não apresentaram mais resultados positivos na nPCR, nem soroconverteram; dois continuaram apresentando resultados positivos na nPCR, mas negativos no IB e IDGA e quatro apresentaram-se positivos na nPCR, no IB e no IDGA. Dessa forma, foi possível demonstrar a infecção interespecífica de forma iatrogênica e a ocorrência de transmissão horizontal do lentivírus caprino entre ovinos.

Assuntos

RESUMO

The production performance of a livestock herd can be compromised by various diseases. In sheep, maedi-visna (MV) infections, which have a chronic nature, are caused by a virus (maedi-visna virus (MVV)) belonging to the genus Lentivirus of the Retroviridae family. The infection can cause significant economic losses and has considerable health impacts on sheep breeding in production systems. Due to the importance of this disease in sheep flocks, the objective was to conduct a serosurvey of MVV in the states of Ceará (CE), Rio Grande do Norte (RN), Paraíba (PB), and Sergipe (SE). A total of, 3332 serum samples were collected in the four states, 1011 in CE, 931 in RN, 459 in PB, and 931 in SE, with the number of samples proportional to the actual herd size of each state. The samples were analyzed using the agar gel microimmunodiffusion test (AGID). Reproducers were revaluated using western blotting (WB). In addition to this serological survey, we administered an investigative questionnaire to identify possible risk factors that facilitate the introduction and spread of diseases (location, category, sex, breed type, creation system, production, herd size, and association with goats). After analysis of the sera using the AGID test, there was zero prevalence. Revaluating breeders by WB revealed a 5.5% prevalence of MV in the four states studied, with prevalences for the states of CE, RN, Paraiba, and SE of 2.3% (2/88), 10.4% (8/77), 3.6% (1/28), and 4.7% (2/42), respectively, corresponding to 13 breeders containing antibodies to the virus. These findings emphasized that the choice of diagnostic tests is extremely important for the early detection of seropositive animals and thus the prevention of the spread of the virus among herds in the region.

O desempenho produtivo de um rebanho pode ser comprometido por diversas enfermidades. Em relação aos ovinos, à ocorrência da Maedi - Visna (MV) doença infecciosa, de caráter crônico, causada por um vírus (Maedi-Visna Vírus MVV) pertencente ao gênero Lentivirus da família Retroviridae. A infecção pode causar importantes perdas econômicas e elevados impactos sanitários nos sistemas de produção da ovinocultura. Em virtude da importância desta enfermidade nos rebanhos ovinos, o objetivo do estudo foi realizar um levantamento soroepidemiológico do vírus da Maedi - Visna (MVV) nos estados do Ceará (CE), Rio Grande do Norte (RN), Paraíba (PB) e Sergipe (SE). Para tanto, foram coletadas 3332 amostras de sangue nos quatro estados, sendo 1011 do CE, 931 do RN, 459 da PB e 931 de SE, sendo este número de amostras proporcional ao rebanho efetivo de cada estado. As amostras foram analisadas utilizando o teste de microimunodifusão em gel de ágar (IDGA). Os reprodutores foram reavaliados pela técnica de Western Blot (WB). Associado a esse levantamento sorológico foi aplicado um questionário epidemiológico para identificar possíveis fatores de risco que podem facilitar a introdução e disseminação de enfermidades (localização, categoria, sexo, tipo racial, sistema de criação, produção, tamanho de rebanho e associação com caprinos). Após análise dos soros pelo teste de IDGA foi verificada uma soroprevalência nula nos rebanhos estudados. Os reprodutores reavaliados pelo WB apresentaram uma prevalência geral de 5,5% da MV nos quatro estados estudados, sendo que os estados do Ceará, Rio Grande do Norte, Paraíba e Sergipe apresentaram, 2,3% (2/88), 10,4% (8/77), 3,6% (1/28) e 4,7% (2/42) respectivamente, correspondendo a 13 reprodutores que apresentaram presença de anticorpos. Diante desses resultados, ressalta-se que a escolha dos testes diagnósticos é de extrema importância para que haja uma detecção precoce de animais soropositivos, e assim evitar a...

Assuntos