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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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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...

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Background: Goats can be infected by a lentivirus called the caprine arthritis encephalitis virus (CAEV) that causes an infectious disease characterized by a chronic. For replication, the CAEV, integrate as a provirus in the DNA of the host cell genome. By consequence, infection of cells persistent life-long infection of the animal. The CAEV can be found in most body fluids, It has been demonstrated its presence in blood, milk, semen. However, the search for the CAEV in other body fluids besides blood is important to assess possible viral transmission. The aim of this study was to determine whether the presence of proviral DNA sequences in saliva of animals infected.Materials, Methods & Results: The study was carried out on the farm belonging to the Embrapa Goat and Sheep Research Center, located in the municipality of Sobral, Ceará, Brazil. To assess the oral fluid for the presence of CAEV, samples of saliva from eight infected breeders were collected by suctioning saliva from the oral cavity on the side region of the breeders lower molar teeth using a probe coupled to a plastic 5 mL syringe. And pro-viral DNA was extracted from the samples using NaCl and proteinase K. Two rounds of polymerase chain reaction (nested PCR) were carried out to amplify the final 187 pb fragment of the pro-viral DNA. All the oligonucleotide primers were determined from the gag region...(AU)

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Background: Goats can be infected by a lentivirus called the caprine arthritis encephalitis virus (CAEV) that causes an infectious disease characterized by a chronic. For replication, the CAEV, integrate as a provirus in the DNA of the host cell genome. By consequence, infection of cells persistent life-long infection of the animal. The CAEV can be found in most body fluids, It has been demonstrated its presence in blood, milk, semen. However, the search for the CAEV in other body fluids besides blood is important to assess possible viral transmission. The aim of this study was to determine whether the presence of proviral DNA sequences in saliva of animals infected.Materials, Methods & Results: The study was carried out on the farm belonging to the Embrapa Goat and Sheep Research Center, located in the municipality of Sobral, Ceará, Brazil. To assess the oral fluid for the presence of CAEV, samples of saliva from eight infected breeders were collected by suctioning saliva from the oral cavity on the side region of the breeders lower molar teeth using a probe coupled to a plastic 5 mL syringe. And pro-viral DNA was extracted from the samples using NaCl and proteinase K. Two rounds of polymerase chain reaction (nested PCR) were carried out to amplify the final 187 pb fragment of the pro-viral DNA. All the oligonucleotide primers were determined from the gag region...

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The aim of this study was to investigate the possible effects of reproductive experience on dopaminergic profile in three different brain tissues, hypothalamus, striatum and cortex in rats on 7th-8th day of pregnancy during the light-dark shift (between 1700-1900h). Results showed that in hypothalamus, dopamine levels increased and DOPAC/DA decreased as a function of parity. In cortex, no differences were observed. In striata, the haloperidol-induced HVA and HVA/DA increases were less intense in experienced animals. These findings suggested that reproductive experience produced functional central changes during pregnancy, with different neurochemical responses depending on the brain region.

A dopamina age nos níveis endócrino, neuroquímico e comportamental. A experiência reprodutiva modula alguns destes aspectos: a dopamina está aumentada no estriato e no hipotálamo de ratas muiltigrávidas entre 12:00-14:00 horas. A sensibilidade dos terminais dopaminérgicos também parece ser modulada por uma experiência reprodutiva prévia. Nosso objetivo foi o de investigar os possíveis efeitos da experiência reprodutiva no perfil dopaminérgico de três diferentes tecidos cerebrais, hipotálamo, estriato e córtex em ratas no 7º-8º dia de gestação durante a fase de virada do ciclo claro-escuro (entre 17:00-19:00h). Nossos resultados mostraram que os níveis dopaminérgicos no hipotálamo aumentaram e o índice DOPAC/DA diminuiu em multigrávidas quando comparadas às primigrávidas. No córtex, nenhuma diferença foi encontrada. No estriato, o aumento de HVA e HVA/DA induzidos pelo haloperidol foi menos intenso nos animais experientes. Nossos resultados sugerem que a experiência reprodutiva produz alterações funcionais centrais durante a gestação, com diferentes respostas neuroquímicas de acordo com a região cerebral.

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Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and was ranked as the most stable in abomasum and lymph node tissues. On the other hand, SDHA was the most stable in the small intestine followed by RPLO and GAPDH. These findings demonstrate the importance of correctly choosing reference genes prior to relative quantification. In addition, we determined that reference genes used in sheep from temperate regions, when properly tested, can be applied in animals from tropical regions such as the Brazilian Somalis sheep.

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Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and was ranked as the most stable in abomasum and lymph node tissues. On the other hand, SDHA was the most stable in the small intestine followed by RPLO and GAPDH. These findings demonstrate the importance of correctly choosing reference genes prior to relative quantification. In addition, we determined that reference genes used in sheep from temperate regions, when properly tested, can be applied in animals from tropical regions such as the Brazilian Somalis sheep.

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