In vivo evaluation of the antiretroviral activity of Melia azedarach against small ruminant lentiviruses in goat colostrum and milk.

This study aimed to evaluate in vivo the use of the extract from the leaves of Melia azedarach in the ethyl acetate fraction at a concentration of 150 µg/mL as an antiretroviral treatment against small ruminant lentiviruses (SRLV) in goat colostrum, and milk with a 90-min action. Two groups of six kids were treated with the extract. One group received three supplies of colostrum from does naturally positive for SRLV, treated with the ethyl acetate fraction of M. azedarach (EAF-MA) for three days, while the other group consumed milk from does also carrying the virus with the respective extract twice a day for five days. After undergoing treatment, all animals began to receive thermized milk until weaning (60 days) and were monitored for six months using nested polymerase chain reaction (nPCR) and western blot (WB) tests. The study revealed cumulative percentages of positive animals in WB or nPCR in the milk group of 66.66% on the seventh day, 83.33% in the following week, and 100% at 120 days, while the colostrum group showed values of 66.66% at 14 days, 83.33% at 90 days, and 100% at 120 days. Variation and intermittency were observed in viral detection, but all animals tested positive in WB or nPCR at some point. A potential delay in infection was observed, which was more significant in the colostrum group. The need for the combination of serological and molecular tests for a more efficient detection of the disease is also emphasized.

Risk factors associated with Leptospira sp. seroprevalence in goats in Northeast of Brazil.

Goat farming is of great importance for agriculture in Northeast of Brazil, which owns more than 90 % of the national herd, however, most studies on leptospirosis in goats in this region are local surveys with unplanned sampling. Thus, the objective of this study was to determine the factors associated with the seroprevalence of leptospirosis in goats in Northeast region of Brazil. Blood samples were collected from 4121 goats from 222 herds, belonging to the states of Ceará, Paraíba, Piauí , Rio Grande do Norte and Sergipe. Serological diagnosis was carried out using the microscopic agglutination test (MAT). In 82.9 % (95 % CI = 77.3-87.2 %) of the herds there was at least one animal seropositive for at least one of the serogroups of Leptospira sp., and 18.73 % (95 % CI = 17.5-19.9 %) of the animals were seropositive. Antibody titers ranged from 100 to 800 and the most frequent serogroups were Autumnalis (23.06 %) and Australis (20.98 %). According to the robust Poisson regression analysis, four factors were associated with seroprevalence: animal sex (female) (P = 0.040), educational level of the farmer (elementary school) (P < 0.001), contacting institution for technical support (P = 0.040) and presence of irrigation area (P = 0.002). There was a high frequency of Leptospira sp. in goats in the states of Northeast region of Brazil, and it's suggested the need for better technological and preventive assistance of herds.

Detection and isolation of small ruminant lentivirus in the amniotic fluid of goats.

The objective of this study was to verify the presence of small ruminant lentivirus in the amniotic fluid of goats using molecular tests and viral isolation by cocultivation in the amniotic fluid of naturally infected goats. The study analyzed eight goats: seven were small ruminant lentivirus-positive and one was negative. The amniotic fluid was collected from each of the eight animals during cesarean section at 147 days of pregnancy. Cocultivation was undertaken using secondary goat nictitating membrane cell cultures obtained by explant from a small ruminant lentivirus-negative calf followed by trypsinization and sub-cultivation of the cells for 63 days. During this period, five supernatant collections were performed for DNA extraction and subsequent nested polymerase chain reaction. DNA was extracted from the amniotic fluid after 3 h of cellular sedimentation, from which a sample of 600 µL was taken from the sediment and another 600 µL sample from the supernatant. After DNA extraction, nested polymerase chain reaction was performed. Of the eight goats, 62.5 % (05/08) were small ruminant lentivirus-positive, with 43.75 % (07/16) of the total samples positive when considering the two repetitions (supernatant and cell sediment). Moreover, positivity was confirmed by small ruminant lentivirus pro-viral DNA amplification in the cell supernatant throughout the cocultivation period. Small ruminant lentivirus were present in the amniotic fluid samples from the naturally infected goats indicating an intrauterine transmission route. Moreover, this biological fluid can be adopted for the diagnosis of these lentiviruse because it is an important risk factor related to intrauterine transmission.

Spatial distribution of serologically reactive sheep to Leptospira spp. in the northeast region of Brazil

Considering the importance of leptospirosis in sheep farming and public health and the significance of identifying which serogroups circulate in sheep within each region, the objective of this study was to determine the seroprevalence and spatial distribution of the most frequent serogroups causing infection by Leptospira sp. in ovine herds in the Northeast region of Brazil. Blood samples were collected from 4197 sheep from 229 herds in 7 Northeastern States. Sera were analyzed via microscopic agglutination test (MAT). The frequency of seroreactive sheep for Leptospira sp. was 14.06%. The states of Alagoas, Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe, located in the Caatinga biome, had the highest frequencies of serologically reactive sheep, and Maranhão, in the Cerrado biome, had the lowest frequency. The most frequent serogroups were Autumnalis (19.49%), Australis (15.76%) and Serjoe (14.41%). In the states of Ceará, Paraíba, Rio Grande do Norte, and Sergipe, 100% of their municipalities had at least one seroreactive animal. The highest frequencies of seropositive animals were found in the municipalities of União (50%), Passagem (49.06%), Canindé (48.89%), Igaci (28.95%), Gararu (31.2%), Pirapemas (17.5%), and Angicos (16%) located in the states of Piauí, Paraíba, Ceará, Alagoas, Sergipe, Maranhão and Rio Grande do Norte, respectively. The animal-level prevalence (14.06%) obtained in the present study is significant, especially considering the rustic nature of the species and the adverse conditions of the region for the infectious agent. In semi-arid conditions, it has been suggested that perhaps sheep do not seroconvert detectable titers on MAT with a cut-off point of 1:100. It is important to highlight that the ovine population in the Northeast region of Brazil is composed of mixed animals, which have been considered more resistant to infection by Leptospira spp. Also, environmental factors hostile to the survival of the infectious agent in the studied region should be taken into consideration, since they may have influenced the seropositive animal-level prevalence. A noteworthy variation was observed in agglutinin titers, which ranged from 100 to 1,600, where 80.2% of the positive samples had titers ≤ 200. It is important to highlight that more elevated titers (≥ 400) were obtained in all seven states, which may suggest an acute infection caused by a non-adapted serovar, indicating that preventive and control measures focused on possible infection sources for sheep should be adopted. Although some states showed the same serogroups as the most frequent, a variety of serogroups was observed in municipalities, which may indicate different sources of infection, whether interspecies, intraspecies, or via alternative routes of transmission in semi-arid conditions, such as venereal. This indicates that even though sheep are more resistant to infection, they become exposed due to the environment or management conditions. As such, identification, isolation, and treatment of the affected animals are alternative measures recommended for prevention and control of leptospirosis in sheep in the semi-arid region. It is evident that despite the lack of rain observed in the last decade in the Northeast region of Brazil, which prevented the formation of favorable environments for the presence of Leptospira, the infectious agent remains among the sheep, as well as other production and wild animals in the region. Some factors may be contributing to this scenario, such as the fact that sheep farming in the region is = characterized mainly by subsistence systems, where veterinary assistance and adequate sanitary management are absent, thus increasing the possibility of contact with Leptospira. (AU)

Vertical transmissibility of small ruminant lentivirus.

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.

Factors associated with the seroprevalence of caseous lymphadenitis in sheep from Northeastern Brazil.

Caseous lymphadenitis (CL) is one of the main infectious diseases in sheep, responsible for great economic losses, mainly in Northeast region of Brazil, which has the largest sheep flock in the country. Thus, the aim of this survey was to determine the factors associated with the seroprevalence of this disease in sheep from Northeastern Brazil using a planned sample of flocks and animals. Samples were collected from 2312 adult and young sheep of both sex from 196 farms, located in 51 municipalities in five Northeastern Brazilian states (Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe). Serological diagnosis was performed using the indirect ELISA technique. Factors associated with seropositivity were identified by variables extracted from epidemiological questionnaires administered to breeders, using univariable and multivariable analyses. Factors associated with prevalence were purebred sheep (Prevalence ratio - PR = 1.189; P = 0.017), ram acquisition from animal expositions (PR = 1.192; P = 0.020), offspring is separated from ewes (PR = 1.132; P = 0.048), water supplied to sheep from ponds (PR = 1.365; P = 0.002), and delayed disposal of infected animals (PR = 1.263; P = 0.027). Flock-level and animal-level prevalences were 93.88 % (95 % CI = 89.60-96.46%) and 37.46 % (95 % CI = 35.51-39.45%), respectively. In view of the high prevalence found here and based on the associated factors, the implementation of an efficient CL control programme in Northeastern Brazil is recommended.

Seroprevalence of Brucella ovis-epididymitis, smooth-Brucella, leptospirosis, toxoplasmosis, and Maedi-Visna in sheep slaughtered in Minas Gerais State, Brazil / Soroprevalência de Brucella ovis, Brucella lisa, leptospirose, toxoplasmose e Maedi-Visna em ovinos abatidos em Minas Gerais, Brasil

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-Leptospiraantibodies, 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, Leptospiraspp., 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% of the sheep exhibited simultaneously antibodies against more than one pathogen. Thus, data from the present study on sheep slaughtered in Minas Gerais, Brazil, showed no antibodies to smooth-Brucella and a low frequency of antibodies anti-Maedi Visna lentivirus, and a high and widespread seroprevalence of B. ovis, Leptospira spp., and T. gondii among animals and flocks.(AU)

O presente estudo teve como objetivo estimar a prevalência de anticorpos contra Brucella ovis (epididimite ovina), Brucellalisa, 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 animais expostos a quatro ou mais dos agentes estudados. No entanto, apenas 25,47% dos ovinos exibiram simultaneamente anticorpos contra mais de um patógeno. Assim, os dados do presente estudo sobre ovinos abatidos em Minas Gerais, Brasil, mostram que ausência de anticorpos anti-Brucella lisa e baixa frequência de anticorpos anti-Maedi Visna, e uma soroprevalência alta e generalizada de B. ovis, Leptospira spp. e T. gondii entre animais e rebanhos.(AU)

Seroprevalence and associated risk factors of Mycoplasma agalactiae and investigation of coinfection with the caprine lentivirus in Rio Grande do Norte, Brazil.

Contagious agalactia is a disease caused by Mycoplasma agalactiae that leads to a reduction or complete stop of milk production. Caprine arthritis encephalitis (CAE) is an infectious disease caused by a lentivirus of the Retroviridae family, member of the small ruminant lentivirus (SRLV) group. Although these diseases are caused by distinct pathogens, the clinical presentation is similar. Hence, this study aimed to perform a serological investigation, as well as to assess correlation between both diseases and risk factors associated in two mesoregions of Rio Grande do Norte, Brazil. Enzyme-linked immunosorbent assay (ELISA) was used for contagious agalactia and western blot for CAE. A total of 538 serum samples were used in this study that were collected from goats and sorted from a blood bank of the Brazilian Agricultural Research Corporation. Seroprevalence of M. agalactiae in flocks from Rio Grande do Norte was 7.8% (42/538). In both regions that were investigated, 25.9% (14/54) of farms had positive animals. CAE results revealed that 3.9% (21/538) of animals and 42.6% (23/54) of farms had this disease. Concerning risk factors, only sex and animal category presented significant relevance (P < 0.05) for contagious agalactia, in which females presented higher frequency of seropositive individuals (10.1%; 39/387). In the animal category, 4.3% (14/326) and 11.1% (36/323) of female breeders were positive for CAE and contagious agalactia, respectively, and significance was identified only in the latter (P < 0.05). In conclusion, there was no correlation between the investigated diseases, considering that no animal demonstrated antibodies for both pathogens.

Seroepidemiological study of Caseous lymphadenitis in sheep from the Northeast region of Brazil using an indirect ELISA.

The objective of this study was to determine the frequency of seropositivity Corynebacterium pseudotuberculosis in sheep in five states of northeastern Brazil, using an indirect enzyme-linked immunosorbent assay (i-ELISA). Young and adult sheep of both sexes were used. Blood samples were collected from 2638 sheep from 223 herds across all states. For the i-ELISA, antigens produced from the strain of C. pseudotuberculosis BRM 029971, a bacterial isolate from the Northeast region of Brazil, were used. Sensitivity and specificity indexes were calculated for the validation of the test, using as reference 49 and 134 serum samples from sheep known to be positive and negative, respectively. The i-ELISA presented four false-negative and four false-positive results, showing a specificity of 97.01%, a sensitivity of 91.84%, and an accuracy of 95.63%. These results were calculated based on an optical density (OD) cutoff point = 0.138. Of the 2638 sheep tested, 996 (37.76%, 95% CI = 35.93-39.62%) were seropositive, and of the 223 evaluated herds, 210 (94.17%, 95% CI = 90.28-96.56%) seropositive. The i-ELISA showed adequate sensitivity and specificity, proving to be a useful tool in the diagnosis of caseous lymphadenitis (CLA) in sheep. Infection by C. pseudotuberculosis, determined by serology, is disseminated in the sheep herds in the states of Northeast Brazil. Thus, there is a need to implement effective control measures that prevent the spread of infectious agents.

Seroprevalence of Corynebacterium pseudotuberculosis and Toxoplasma gondii in sheep in semi-arid region of Ceará state / Soroprevalência de Corynebacterium pseudotuberculosis de Toxoplasma gondii em ovinos de região semi-árida do estado do Ceará

ABSTRACT: The present study was conducted to investigate in 20 extensive sheep farms for the seroprevalence of Corynebacterium pseudotuberculosis (n=402) and Toxoplasma gondii (n=228). Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to C. pseudotuberculosis/T. gondii. It was observed that C. pseudotuberculosis showed the highest prevalence in the region (34.07%) with statistically significant presence (p<0.05) in ewes. Antibodies to T. gondii was reported in 14.91% of the animals studied. About C. pseudotuberculosis/T. gondii coinfection the categories of rams showed significant (p<0.05) differences, suggesting that this gender could perpetuate the diseases in the flocks. It was concluded that the knowledge about the diseases in the region under study would facilitate the execution of prophylactic measures, especially against the diseases that pose risks to the public health and cause damages to the producer.

RESUMO: O presente estudo foi conduzido para investigar a soroprevalência em 20 fazendas de criação extensiva de ovinos quanto à presença de anticorpos para Corynebacterium pseudotuberculosis (n=402) e Toxoplasma gondii (n=228). Ensaio de imunoabsorção enzimático (ELISA) foi utilizado para a detecção de anticorpos contra C. pseudotuberculosis/T. gondii. Observou-se que C. pseudotuberculosis apresentou a maior prevalência na região (34,07%), com presença estatisticamente significante (p <0,05) nas categorias de ovelha. Anticorpos contra T. gondii foram encontrados em 14,91% dos animais estudados. Sobre a coinfecção de C. pseudotuberculosis/T. gondii, as categorias carneiro apresentaram diferenças significativas (p <0,05), sugerindo que esse gênero poderia perpetuar as doenças nos rebanhos. Concluiu-se que o conhecimento sobre as doenças na região em estudo facilitaria a execução de medidas profiláticas, principalmente contra as doenças que apresentam riscos à saúde pública e causam danos ao produtor.

Seroepidemiological characterization and risk factors associated with seroconversion to Corynebacterium pseudotuberculosis in goats from Northeastern Brazil.

Goat breeding in the Northeast region of Brazil plays an important socioeconomic role. However, there are significant losses caused by sanitary deficits and infectious diseases, particularly caseous lymphadenitis (CL). Although CL is considered endemic in Northeastern Brazil, a comprehensive and up-to-date study of this disease in goat herds in this region is necessary. The objective of this study was to determine the farm-level and animal-level seroprevalences for the disease and to identify the possible risk factors that characterize CL in the caprine species of five Northeastern's states (Ceará, Piauí, Rio Grande do Norte, Paraíba, and Sergipe). A total of 2744 goat serum samples from 230 farms were collected between 2010 and 2012. The diagnosis of Corynebacterium pseudotuberculosis infection was performed using the indirect ELISA technique. Farm-level and animal-level seroprevalences were 87.8% and 30.3%, respectively, suggesting that C. pseudotuberculosis is widespread in goat herds of the Northeast region. The risk factors were as follows: absence of forage silage (odds ratio = 5.39), not separating animals by sex (odds ratio = 4.16) or by age (odds ratio = 6.30), not replacing old goat breeders (odds ratio = 7.80), and non-treatment of CL lumps prior to spontaneous rupture (odds ratio = 10.34). This study supports the idea that caseous lymphadenitis is widely disseminated in goats from Northeastern Brazil and based on the risk factor analysis attention should be given to the need to establish adequate control measures, such as incision and early drainage of superficial abscesses, quarantine and elimination of affected animals, periodic inspection of the herd, non-introduction of infected animals, and early disposal of animals with recurrent CL.

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

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

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

A panel of protein candidates for comprehensive study of Caprine Arthritis Encephalitis (CAE) infection.

The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.

Serum epidemiological survey and risk factors investigation for lentivirus in goats from Sisal Region, Bahia, Brazil / Inquérito soroepidemiológico e fatores de risco para lentivírus em rebanhos caprinos da região sisaleira, Bahia

This study aimed to carry out a serum epidemiological survey of goat arthritis encephalitis in the sisal region of Bahia, Brazil, and to evaluate risk factors. We evaluated 831 samples of goat blood serum among males and females older than six months, from 49 farms distributed among the municipalities of Araci, Cansanção, Conceição do Coité, Itiúba, Monte Santo, Nordestina, Queimadas, Santa Luz, São Domingos and Valente. An epidemiological questionnaire for the analysis of possible risk factors was applied. All sera were tested for immune-agar gel. The seroprevalence obtained in surveyed herds was 1.56% (13/831). There was significant difference (p<0.05) for animal racial pattern, type of farming and breeding systems. However, when considering herds with predominantly dairy breeds (Saanen and Alpine Pardo), the seropositivity in animals from Valente, Conceição do Coité and São Domingos amounted to 5.06% (12/237). In these municipalities, from 14 properties analyzed, five (38.5%) had at least one positive-testing animal. This result is extremely worrying when one considers that few control measures are adopted by farmers to prevent the goat arthritis encephalitis in the most important dairy region of Bahia state.(AU)

Este trabalho teve por objetivo realizar um levantamento soroepidemiológico da artrite encefalite caprino na região sisaleira do estado da Bahia e avaliar os fatores de risco. Foram avaliadas 831 amostras de soros sanguíneos de caprinos entre machos e fêmeas, com idade superior a seis meses, provenientes de 49 propriedades rurais distribuídas entre os municípios de Araci, Cansanção, Conceição do Coité, Itiúba, Monte Santo, Nordestina, Queimadas, Santa Luz, São Domingos e Valente. Foi aplicado um questionário epidemiológico destinado à análise de possíveis fatores de risco. Todos os soros foram submetidos ao teste da imunodifusão em gel de ágar. A soroprevalência obtida nos rebanhos pesquisados foi de 1,56% (13/831). Houve diferença significativa (p≤0,05) para padrão racial dos animais, tipo de exploração e sistemas de criação. Quando se consideraram apenas os rebanhos com raças predominantemente leiteiras (Saanen e Pardo alpina), dos municípios de Valente, Conceição do Coité e São Domingos, a soropositividade nos animais elevou-se para 5,06% (12/237). Nesses municípios, de 14 propriedades analisadas, 5 (38,5%) apresentaram pelo menos um animal sororeagente. Esse resultado é extremamente preocupante quando se constata que poucas medidas de controle são adotadas pelos criadores para impedir a disseminação dessa importante enfermidade na bacia leiteira mais relevante do estado da Bahia.(AU)

Transmission of caprine arthritis encephalitis virus between sheep / Transmissão do vírus da artrite encefalite caprina entre ovinos

ABSTRACT: This study was conducted to evaluate caprine arthritis encephalitis virus (CAEV) transmission among sheep using 15 lambs that were distributed in 2 experimental groups. The exposed group consisted of 10 lambs that remained with their mothers, who were experimentally infected with CAEV. The non-exposed group was characterized as the control group and was comprised of 5 lambs that remained with their CAEV-negative mothers. Blood samples were collected monthly from birth until 1 year of life. To evaluate the transmission, an agar gel immunodiffusion test (AGID), enzyme immunoassay (ELISA), immunoblotting (IB), and nested polymerase chain reaction (nPCR) techniques were used. The non-exposed group was negative in all of the tests throughout the whole experiment. In the exposed group, 2 individuals had positive nPCR results. Positive nPCR samples were sequenced for comparison with the original goat strains and were shown to be similar to the CAEV-Cork strain. Seroconversion was not detected, and clinical manifestations were not observed. Thus, after 1 year of observation, it was verified that CAEV transmission among sheep is possible; however, with discreet frequency. This was an initial study, and other experiments are needed to analyze the adaptive capacity of the CAEV to remain in an infected sheep flock and cause the disease.

RESUMO: O estudo foi conduzido para avaliar a transmissão do vírus da artrite encefalite caprina (CAEV) entre ovinos, utilizando 15 cordeiros, distribuídos em dois grupos experimentais. O grupo exposto foi constituído por 10 cordeiros, mantidos com suas mães, que foram infectadas, experimentalmente, com CAEV. O grupo não exposto caracterizou-se como grupo controle e foi formado por cinco cordeiros, mantidos com suas matrizes, negativas para CAEV. Foram colhidas amostras de sangue mensalmente, do periodo que compreende o nascimento até um ano de vida. Para avaliar a transmissão, foram utilizadas as técnicas de imunodifusão em gel de agarose (IDGA), ensaio imunoenzimático (ELISA), immunoblotting (IB) e reação em cadeia da polimerase do tipo nested (nPCR). O grupo não exposto se manteve negativo aos testes durante todo o experimento. Já no grupo exposto, dois indivíduos apresentaram resultados positivos na nPCR. As amostras positivas na nPCR foram sequenciadas para serem comparadas com as cepas originais de caprinos, comprovando se tratar de lentivírus semelhante à cepa CAEV-Cork. A soroconversão não foi detectada e a manifestação clínica não foi observada. Sendo assim, após um ano de observação, verificou-se que a transmissão do CAEV entre ovinos é possível, entretanto, com discreta frequência. Este foi um estudo inicial, e outros experimentos são necessários para analisar a capacidade adaptativa do CAEV de permanecer em rebanho ovino infectado e, com isso, causar doença.

Small ruminant lentiviruses: economic and productive losses, consequences of the disease / Lentiviroses de pequenos ruminantes: perdas produtivas e econômicas, consequências da doença

Small ruminant lentiviruses, caprine arthritis encephalitis virus, and Maedi-Visna virus cause diseases that result in significant productive losses, mostly in dairy animals. These viruses belong to the Retroviridae family, Lentivirus genus, and constitute a heterogeneous group, which may generate implications for the diagnosis and control of small ruminant lentiviruses. Losses caused by them are associated with reproductive failure, short productive life, and decreased milk production by the infected animals. In addition, these viruses may reduce milk quality, affecting the production of dairy products such as cheese. Small ruminant lentiviruses lead to indirect losses, decreasing herd value and forcing the development of epidemiological trade barriers for animal germplasm. Control of small ruminant lentiviruses is important to promote optimal milk production and to reduce costs with medicine and technical assistance. This control may vary in caprine and ovine populations of each country, according to seroprevalence, variety of breeds, and peculiarities of the practiced management.(AU)

Os lentivírus de pequenos ruminantes, o vírus da artrite encefalite caprina e o vírus Maedi-Visna causam enfermidades que ocasionam perdas produtivas significativas, principalmente em animais com aptidão leiteira. Esses vírus pertencem à família Retroviridae e ao gênero Lentivirus e formam um grupo genético heterogêneo, o que pode ocasionar implicações para o diagnóstico e o controle dos lentivírus de pequenos ruminantes. As perdas causadas pelos lentivírus de pequenos ruminantes estão relacionadas com falhas reprodutivas, vida produtiva curta e diminuição da produção leiteira dos animais infectados. Além disso, esses vírus podem promover a redução da qualidade do leite, afetando a produção de laticínios, tal como o queijo. Os lentivírus de pequenos ruminantes levam a perdas indiretas, reduzindo o valor dos rebanhos e forçando o desenvolvimento de barreiras comerciais epidemiológicas para germoplasma animal. O controle dos lentivírus de pequenos ruminantes é importante para promover uma maior produção de leite e reduzir os custos com medicamentos e assistência técnica. Esse controle pode variar de acordo com a população caprina e ovina de cada país em termos de soroprevalência, variedade de raças e particularidades do manejo adotado.(AU)

Evaluation of caprine arthritis-encephalitis virus transmission in newborn goat kids / Estudo da transmissão do vírus da artrite-encefalite caprina em cabritos neonatos

Caprine arthritis encephalitis causes considerable losses in goat production. The main form of the caprine arthritis encephalitis virus transmission is through the ingestion of colostrum or milk from infected females. However, some transmissions cannot be explained in this manner. Therefore, this study aimed to evaluate transplacental transmission of caprine arthritis encephalitis virus. Blood samples were collected from 283 newborn kids of Anglo-Nubian and Saanen breeds born from seropositive and seronegative goats. Samples were collected immediately after birth and analyzed with agarose gel immunodiffusion and western blot. All samples were negative in the agarose gel immunodiffusion. However, the western blot test demonstrated that four kids were born positive for caprine arthritis encephalitis virus. This result indicates that although in a low frequency (1.4%), there is a possibility of transplacental transmission of small ruminant lentivirus.(AU)

A artrite encefalite caprina causa perdas consideráveis para a produção caprina. A principal forma de transmissão do vírus da artrite encefalite caprina é a ingestão de colostro ou leite de fêmeas infectadas. No entanto, algumas transmissões não podem ser explicadas por esta via. Dessa forma, este estudo teve como objetivo avaliar a transmissão do vírus da artrite encefalite caprina por via transplacentária (vertical). Foram realizadas coletas de sangue em 283 crias recém-nascidas das raças Anglo-Nubiana e Saanen, provenientes de progenitores soropositivos e soronegativos. As amostras foram coletadas logo após o nascimento e analisadas pelas técnicas de imunodifusão em gel de agarose e western blot. No teste de imunodifusão em gel de agarose, nenhum cabrito foi detectado reagente. Porém, no teste de western blot, quatro cabritos nasceram soropositivos. Esse resultado indica que, apesar de baixa frequência (1,4%), existe a possibilidade de transmissão via transplacentária do lentivírus de pequenos ruminantes.(AU)

Investigação sorológica das lentiviroses de pequenos ruminantes nas microrregiões homogêneas do Alto Médio Canindé, Picos e Floriano, Piauí, Brasil / Serological investigation of lentiviruses of small ruminants in the microregions of Alto Médio Canindé, Picos and Floriano, Piauí state, Brazil

Lentivírus de pequenos ruminantes (LV) é o termo genérico utilizado para designar os vírus da artrite encefalite caprina e Maedi-Visna, os quais pertencem à família Retroviridae, subfamília Orthoretrovirinae, gênero Lentivirus. Tais vírus infectam caprinos e ovinos, causando enfermidades de curso lento com lesões inflamatórias, crônicas e degenerativas que podem atingir vários órgãos, provocando caquexia e morte. Os animais infectados eliminam o vírus sobretudo por meio de secreções e excreções e transmitem-no especialmente em situações de estreito contato. Não há tratamento até o momento. O controle é baseado na criação segregada, no manejo e no sacrifício dos positivos. Esse agente infeccioso já foi relatado em várias partes do mundo, sendo responsável por perdas econômicas significativas. Por o agente ter sido verificado em vários estados do Brasil e por não existirem dados soroepidemiológicos nas mesorregiões sudeste e sudoeste piauiense, esta pesquisa teve por objetivo realizar inquérito sorológico para investigar a ocorrência de anticorpos para o LV em ovinos e caprinos nas microrregiões do Alto Médio Canindé, Picos e Floriano, no Piauí. Para tanto, foram coletadas 1.280 e 1.360 amostras de soro caprino e ovino, respectivamente, oriundos de 20 municípios, distribuídos nas três microrregiões, sendo o número de amostras proporcional ao rebanho efetivo de cada município. As amostras de soro foram analisadas utilizando o teste de imunodifusão em gel de agarose (IDGA). Nenhum dos soros pesquisados reagiu positivamente, constatando-se soroprevalência nula. Ressalta-se a importância da implantação de um rigoroso programa de controle para que se possa evitar a introdução e/ou a disseminação desse agente infeccioso nessas microrregiões.(AU)

Small ruminant lentiviruses (LV) is the generic term for the caprine arthritis-encephalitis and ­­Maedi-Visna viruses, which belong to the Retroviridae family, Orthoretrovirinae subfamily, Lentivirus genus. The virus infects goats and sheep, causing slow course of disease with inflammatory, chronic and degenerative lesions, which can reach several organs, provoking cachexia and death. Infected animals eliminate the virus mainly through secretions and excretions and transmit it especially in close contact situations. There is no treatment to date. The control is based on segregated creation, management and sacrifice of the positive. This infectious agent has been reported in various parts of the world and is responsible for significant economic losses. It was verified in several states of Brazil and there are seroepidemiological data in southeast and southwest mesoregions of Piauí, Brazil. This research aimed to perform serological survey to investigate the occurrence of antibodies to LV in sheep and goats, in the regions of Alto Médio Canindé, Picos and Floriano. So, 1,280 and 1,360 serum goats and sheep samples, respectively, were collected, coming from 20 municipalities, distributed in the 3 microregions. The number of samples was proportional to the actual herd of each municipality. The samples were analyzed using the agar gel immunodiffusion test. None of the surveyed sera reacted positively, though there is a null seroprevalence. It was emphasized the importance of implementing a rigorous control program in order to prevent the introduction and spread of this infectious agent in these microregions.(AU)

Investigação sorológica das lentiviroses de pequenos ruminantes nas microrregiões homogêneas do Alto Médio Canindé, Picos e Floriano, Piauí, Brasil / Serological investigation of lentiviruses of small ruminants in the microregions of Alto Médio Canindé, Picos and Floriano, Piauí state, Brazil

Small ruminant lentiviruses (LV) is the generic term for the caprine arthritis-encephalitis and ­­Maedi-Visna viruses, which belong to the Retroviridae family, Orthoretrovirinae subfamily, Lentivirus genus. The virus infects goats and sheep, causing slow course of disease with inflammatory, chronic and degenerative lesions, which can reach several organs, provoking cachexia and death. Infected animals eliminate the virus mainly through secretions and excretions and transmit it especially in close contact situations. There is no treatment to date. The control is based on segregated creation, management and sacrifice of the positive. This infectious agent has been reported in various parts of the world and is responsible for significant economic losses. It was verified in several states of Brazil and there are seroepidemiological data in southeast and southwest mesoregions of Piauí, Brazil. This research aimed to perform serological survey to investigate the occurrence of antibodies to LV in sheep and goats, in the regions of Alto Médio Canindé, Picos and Floriano. So, 1,280 and 1,360 serum goats and sheep samples, respectively, were collected, coming from 20 municipalities, distributed in the 3 microregions. The number of samples was proportional to the actual herd of each municipality. The samples were analyzed using the agar gel immunodiffusion test. None of the surveyed sera reacted positively, though there is a null seroprevalence. It was emphasized the importance of implementing a rigorous control program in order to prevent the introduction and spread of this infectious agent in these microregions.

Lentivírus de pequenos ruminantes (LV) é o termo genérico utilizado para designar os vírus da artrite encefalite caprina e Maedi-Visna, os quais pertencem à família Retroviridae, subfamília Orthoretrovirinae, gênero Lentivirus. Tais vírus infectam caprinos e ovinos, causando enfermidades de curso lento com lesões inflamatórias, crônicas e degenerativas que podem atingir vários órgãos, provocando caquexia e morte. Os animais infectados eliminam o vírus sobretudo por meio de secreções e excreções e transmitem-no especialmente em situações de estreito contato. Não há tratamento até o momento. O controle é baseado na criação segregada, no manejo e no sacrifício dos positivos. Esse agente infeccioso já foi relatado em várias partes do mundo, sendo responsável por perdas econômicas significativas. Por o agente ter sido verificado em vários estados do Brasil e por não existirem dados soroepidemiológicos nas mesorregiões sudeste e sudoeste piauiense, esta pesquisa teve por objetivo realizar inquérito sorológico para investigar a ocorrência de anticorpos para o LV em ovinos e caprinos nas microrregiões do Alto Médio Canindé, Picos e Floriano, no Piauí. Para tanto, foram coletadas 1.280 e 1.360 amostras de soro caprino e ovino, respectivamente, oriundos de 20 municípios, distribuídos nas três microrregiões, sendo o número de amostras proporcional ao rebanho efetivo de cada município. As amostras de soro foram analisadas utilizando o teste de imunodifusão em gel de agarose (IDGA). Nenhum dos soros pesquisados reagiu positivamente, constatando-se soroprevalência nula. Ressalta-se a importância da implantação de um rigoroso programa de controle para que se possa evitar a introdução e/ou a disseminação desse agente infeccioso nessas microrregiões.

Ferramentas diagnósticas de Lentivirose de Pequenos Ruminantes: padronização da técnica de ELISA indireto / Diagnostic tools of small ruminant lentiviruses: standardization of indirect ELISA

As Lentiviroses de Pequenos Ruminantes (LVPR) incluem a Maedi-Visna (MV) em ovinos e a Artrite Encefalite Caprina (CAE). Essas enfermidades estão difundidas no mundo e são responsáveis por grandes perdas na produtividade destes animais. Os LVPR são vírus RNA da subfamília Lentivirinae que causam uma infecção persistente, sendo a detecção precoce uma das formas mais eficientes para limitar sua disseminação no rebanho. Visando contribuir com essas questões, este experimento foi realizado na Universidade Federal do Piauí (UFPI) em parceria com a Embrapa Caprinos e Ovinos, com o objetivo de padronizar a técnica de ensaio imunoenzimático indireto e compará-lo com a imunodifusão em gel de agarose no diagnóstico da CAE. Foram utilizadas 696 amostras de soros de caprinos machos e fêmeas oriundas do banco de soros da Unidade de Pesquisa de LVPR do Centro de Ciências Agrárias da UFPI. As amostras foram coletadas no período de janeiro de 2007 a março de 2010. Na padronização, verificou-se que 0,25 µg de proteína/poço, diluição de 1:200 do soro e concentração de 1:3.000 do conjugado anticorpo anti-IgG cabra apresentaram os melhores resultados. O ponto de corte obtido foi de 0,36. Na comparação, o Imunodifusão em Gel de Ágar (IDGA) detectou 128 (18,4%) amostras positivas, e o ELISA indireto (ELISA-i), 259 (37,2%). A sensibilidade e a especificidade do teste ELISA-i com relação ao IDGA foi de 94,5% e 75,7%, respectivamente. Verificou-se maior índice de positividade em caprinos acima de seis meses (p < 0,05), e nos machos obteve-se prevalência de 56,7% em comparação às fêmeas, 35,4%, (p < 0,01).(AU)

The Small Ruminant Lentiviruses (SRLVs) include Maedi-Visna (MV) of sheep and Caprine Arthritis-Encephalitis (CAE). These diseases are widespread and responsible for major production losses regarding sheep and goats. The SRLV is a RNA virus of the subfamily Lentivirus genus that causes persistent infections in goats. Early detection is one of the best ways to limit its spread in the herd. To contribute to these issues, this experiment was conducted at Universidade Federal do Piauí in partnership with Embrapa Goats and Sheep, with the objective of standardizing the technique of indirect ELISA (i-ELISA) and to compare it with Immunodiffusion in Agarose Gel to diagnose Caprine Lentiviruses (LC). Six hundred ninety six serum samples were used from the University Veterinary Hospital, Universidade Federal do Piauí, from January 2007 to March 2010. Standardization showed that 0.25 µg protein/well, a 1:200 dilution of the serum and concentration of 1:3,000 of the conjugated anti-goat IgG presented the best results. It was observed that the Agar Gel Immunodiffusion (AGID) detected 128 (18.4%) positive samples, and ELISA, 259 (37.2%). The sensitivity and specificity of i-ELISA regarding AGID were 94.5% and 75.7%, respectively. A higher prevalence was observed among animals older than six months (p < 0.05). The prevalence among males was of 56.7%, and among females, 35.4% (p < 0.01).(AU)