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.

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

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.

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

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.

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.

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 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.

In vivo evaluation of antiseptics and disinfectants on control of Caseous Lymphadenitis: clinical, haematological, serological and microbiological monitoring / Avaliação in vivo de antissépticos e desinfetantes no controle da Linfadenite Caseosa: acompanhamento clínico, hematológico, sorológico e microbiológico

The objective of this experiment was to evaluate the efficacy of iodine tincture at 10% and sodium hypochlorite at 2.5% applied into the abscess of animals affected by Caseous Lymphadenitis (CL). Eighteen ewes were used, assorted into three groups: one treated with iodine tincture at 10% (IT), another one with sodium hypochlorite at 2.5% (SH) and the last group underwent the conventional treatment (CT). Conventional treatment was based on surgical drainage and chemical cauterization of the lesion with iodine tincture at 10%. Natural rupture of six abscesses from IT group was observed and in five of them the viability of Corynebacterium pseudotuberculosis was confirmed on the lesion place, after rupture of lymph node. As for the SH group, spontaneous rupture was observed in five out of six abscesses treated, and the microorganism was identified on the lesion of five animals, after rupture. In the sixth animal of this group, abscess involution was noticed. A severe swelling was identified in the region of lymph node treated, resulting in wide lesion in animals from groups IT and SH. No difference (p > 0,05) was found in blood parameters due to treatments. As for the serological monitoring of animals, comparative analysis between months within each group showed that months 1, 2, 3 and 4 were different (p 0,05) from month 0, for groups IT and SH. For CT group, there was no difference (p > 0,05) between months 1 to 5 and 0. Then, the application of iodine tincture at 10% or sodium hypochlorite at 2.5% into the abscess of animals affected by CL, at stage in which lesions are detected through inspection, is not effective for its control.

Objetivou-se com este experimento avaliar a eficácia da tintura de iodo a 10% e do hipoclorito de sódio a 2,5% aplicados no interior do abscesso de animais acometidos pela Linfadenite Caseosa (LC). Foram utilizadas 18 fêmeas ovinas, divididas em 3 grupos: o primeiro foi tratado com tintura de iodo a 10% (TI), o segundo com hipoclorito de sódio a 2,5% (HS) e o último grupo foi submetido ao tratamento convencional (CT). O tratamento convencional da doença consistia na drenagem e cauterização química dos abscessos, utilizando-se solução de iodo a 10%. Foi observada a ruptura natural dos seis abscessos do grupo TI, sendo que em cinco deles a viabilidade do Corynebacterium pseudotuberculosis foi confirmada no local da lesão, após a ruptura do linfonodo. Quanto ao grupo HS, a ruptura espontânea foi observada em cinco dos seis abscessos tratados, sendo o microrganismo identificado na lesão dos cinco animais após o rompimento. No sexto animal desse grupo, foi constatada a involução do abscesso. Foi verificado um edema intenso na região do linfonodo tratado com formação de uma lesão de grande extensão nos animais dos grupos TI e HS. Não foi detectada diferença (p > 0,05) nos parâmetros hematológicos em decorrência dos tratamentos avaliados. Quanto ao acompanhamento sorológico dos animais, a análise estatística comparativa entre os meses, dentro de cada grupo, mostrou que os meses 1, 2, 3 e 4 foram diferentes (p 0,05) do mês 0 para os grupos TI e HS. No grupo CT, não houve diferença (p > 0,05) entre os meses de 1 a 5 e o mês 0. Conclui-se que a aplicação de tintura de iodo a 10% ou hipoclorito de sódio a 2,5% no interior do abscesso de animais acometidos pela LC, em estágio no qual as lesões são detectadas por meio da inspeção, não é eficaz para o seu controle.