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1.
Trop Anim Health Prod ; 50(1): 43-48, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29022241

RESUMO

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.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Doenças das Cabras/diagnóstico , Infecções por Lentivirus/veterinária , Proteínas de Plasma Seminal/análise , Animais , Western Blotting/veterinária , Eletroforese em Gel Bidimensional/veterinária , Doenças das Cabras/virologia , Cabras/genética , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/virologia , Masculino , Reação em Cadeia da Polimerase/veterinária , Sêmen/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária
2.
Anim Reprod ; 16(4): 902-913, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-32368270

RESUMO

The Saanen goat breed has been widely explored in breeding programmes; however, there are few reports about the breed's genetic and molecular composition. Thus, this study aimed to characterize the proteomic profile of spermatozoa from Saanen breeding goats. Five breeding animals with proven fertility were selected, the spermatozoa were collected, and the protein was extracted. Subsequently, the proteins were separated and analysed by two-dimensional electrophoresis and mass spectrometry; the proteins were then identified with the SwissProt database. A total of 31 proteins involved in reproduction were identified, including binding proteins on spermatozoa for fusion with the egg, acrosomal membrane proteins, metabolic enzymes, heat shock proteins, cytoskeletal proteins and spermatozoa motility proteins. The characterization of such proteins clarifies the molecular mechanisms of spermatogenesis and the modifications that ensure the success of fertilization.

3.
Acta Vet. Brasilica ; 17(2): 36-43, 2023. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1512957

RESUMO

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


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


Assuntos
Animais , Feminino , Cabras/virologia , Biomarcadores/análise , Lentivirus Ovinos-Caprinos/isolamento & purificação , Metaloproteinases da Matriz/análise , Reação em Cadeia da Polimerase/veterinária , Imunodifusão/métodos
4.
Anim Reprod ; 15(1): 84-92, 2018 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-33365100

RESUMO

This study aimed to develop protocols for the extraction of sperm proteins from Moxotó goats (Capra hircus) and to compare the resulting proteomic maps. The sperm proteins were isolated using an extraction buffer containing 7 M urea and 2 M thiourea, 20 mM DTT, and one of the following detergents: 1% or 4% CHAPS; 1% or 4% SDS; 1% or 4% Triton X-100; or a combination of CHAPS and SDS. The 1-DE and 2-DE profiles of the isolated proteins revealed that the various isolation methods were efficient. Qualitative and quantitative differences in the 1-DE and 2-DE profiles were observed. 2-DE maps indicated that the amount and diversity of proteins visualized depended on the detergent that was used. Furthermore, this work revealed that the combination of detergents increased the resolution of some spots and retained the characteristics of the individual detergents, depending on their concentrations.

5.
Acta sci. vet. (Online) ; 49: Pub. 1784, Jan. 28, 2021. tab
Artigo em Inglês | VETINDEX | ID: vti-29823

RESUMO

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturers instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...(AU)


Assuntos
Animais , Feminino , Ovinos/microbiologia , Infecções por Chlamydia/veterinária , Infecções por Chlamydia/epidemiologia , Zoonoses , Fatores de Risco , Ensaio de Imunoadsorção Enzimática/veterinária , Aborto Animal
6.
Acta sci. vet. (Impr.) ; 49: Pub.1784-2021. tab
Artigo em Inglês | VETINDEX | ID: biblio-1458423

RESUMO

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturer’s instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...


Assuntos
Feminino , Animais , Infecções por Chlamydia/epidemiologia , Infecções por Chlamydia/veterinária , Ovinos/microbiologia , Aborto Animal , Ensaio de Imunoadsorção Enzimática/veterinária , Fatores de Risco , Zoonoses
7.
Semina ciênc. agrar ; 41(06,supl. 2): 3165-3176, 2020. tab, ilus
Artigo em Inglês | VETINDEX | ID: biblio-1501676

RESUMO

Caprine arthritis encephalitis is a lentiviral disease that leads to considerable losses in goat farming. In the acute phase of viral infection, though antiviral antibodies are produced by the host's immune system, they are not sufficient to be detected by serological tests. Acute infections begin with an incubation period, during which the viral genome replicates and host innate responses are initiated. Matrix metalloproteinases (MMPs) are enzymes that play an important role in the physiological and pathological processes of tissue remodeling. The present study aimed to evaluate the expression of MMPs and their activity in the blood serum of male goats experimentally infected with caprine arthritis encephalitis virus (CAEV). Five dairy male goats, aged 3-4 years, were intravenously inoculated with CAEV Cork strain (titer: 105-6 TCID50/mL) after being tested negative for CAEV thrice at consecutive intervals of 30 days using western blot analysis and nested-PCR. The study included three stages: S1 or pre-infection stage; S2 or seroconversion stage, corresponding to the occurrence of first seroconversion; and S3 or post-seroconversion stage, corresponding to 23 weeks after seroconversion. Zymography was performed for the samples using gelatin zymography gels (12.5%), which were subjected to electrophoresis at 170V, 1A, and 300W for 50-70 min. The density of MMP-2 was found to be lower at S1 (1456.20 pixels) than that at S2 and S3 (1943.80 and 2104.40 pixels, respectively) (P < 0.05); and the density of MMP-9 was found to be lower at S3 (133.60 pixels) than that at S1 and S2 (359.60 and 370.60 pixels, respectively). The density of proMMP-2 was low at S1 and S3 (130.45 and 145.20 pixels, respectively). On the other hand, the density of proMMP-9 was statistically different between S1 and S3 (89.22 vs. 415.60 pixels). Both proMMP-2 and proMMP-9 were absent at S2. Thus, MMP-2 and MMP9 exhibited opposite behaviors depending on the stage [...].


A encefalite por artrite caprina é uma doença lentiviral que leva a perdas consideráveis na criação de caprinos. Na fase aguda da infecção viral, embora os anticorpos antivirais sejam produzidos pelo sistema imunológico do hospedeiro, eles não são suficientes para serem detectados por testes sorológicos. As infecções agudas começam com um período de incubação, durante o qual o genoma viral se replica e as respostas inatas do hospedeiro são iniciadas. As metaloproteinases da matriz (MMPs) são enzimas que desempenham um papel importante nos processos fisiológicos e patológicos da remodelação tecidual. O presente estudo teve como objetivo avaliar a expressão de MMPs e sua atividade no soro sanguíneo de reprodutores caprinos infectados experimentalmente pelo vírus da encefalite por artrite caprina(CAEV). Cinco machos caprinos, com idades entre 3-4 anos, foram inoculadas por via intravenosa com a cepa de CAEV Cork (título: 105-6 TCID50/mL) após serem testados negativamente para CAEV três vezes em intervalos consecutivos de 30 dias usando a análise de western blot e nested-PCR. O estudo incluiu três etapas: estágio S1 ou pré-infecção; S2 ou estágio de soroconversão, correspondente à ocorrência de primeira soroconversão; e S3 ou pós-soroconversão, correspondendo a 23 semanas após a soroconversão. A zimografia foi realizada para as amostras utilizando gel de gelatina (12,5%), que foram submetidos à eletroforese em 170V, 1A e 300W por 50-70 min. Verificou-se que a densidade de MMP-2era menor em S1 (1456,20 pixels) do que em S2 e S3 (1943,80 e 2104,40 pixels, respectivamente) (P <0,05); e a densidade de MMP-9 foi menor em S3 (133,60 pixels) do que em S1 e S2 (359,60 e 370,60 pixels, respectivamente). A densidade do proMMP-2 era baixa em S1 e S3 (130,45 e 145,20 pixels, respectivamente). Por outro lado, a densidade do proMMP-9 foi estatisticamente diferente entre S1 e S3(89,22 vs. 415,60 pixels). ProMMP-2 e proMMP-9 estavam ausentes no S2. Assim, MMP-2 e [...].


Assuntos
Masculino , Animais , Infecções por Lentivirus/enzimologia , Infecções por Lentivirus/sangue , Infecções por Lentivirus/veterinária , Lentivirus Ovinos-Caprinos/patogenicidade , Metaloproteases/sangue , Ruminantes/sangue , Ruminantes/virologia
8.
Semina Ci. agr. ; 41(06,supl. 2): 3165-3176, 2020. tab, ilus
Artigo em Inglês | VETINDEX | ID: vti-26736

RESUMO

Caprine arthritis encephalitis is a lentiviral disease that leads to considerable losses in goat farming. In the acute phase of viral infection, though antiviral antibodies are produced by the host's immune system, they are not sufficient to be detected by serological tests. Acute infections begin with an incubation period, during which the viral genome replicates and host innate responses are initiated. Matrix metalloproteinases (MMPs) are enzymes that play an important role in the physiological and pathological processes of tissue remodeling. The present study aimed to evaluate the expression of MMPs and their activity in the blood serum of male goats experimentally infected with caprine arthritis encephalitis virus (CAEV). Five dairy male goats, aged 3-4 years, were intravenously inoculated with CAEV Cork strain (titer: 105-6 TCID50/mL) after being tested negative for CAEV thrice at consecutive intervals of 30 days using western blot analysis and nested-PCR. The study included three stages: S1 or pre-infection stage; S2 or seroconversion stage, corresponding to the occurrence of first seroconversion; and S3 or post-seroconversion stage, corresponding to 23 weeks after seroconversion. Zymography was performed for the samples using gelatin zymography gels (12.5%), which were subjected to electrophoresis at 170V, 1A, and 300W for 50-70 min. The density of MMP-2 was found to be lower at S1 (1456.20 pixels) than that at S2 and S3 (1943.80 and 2104.40 pixels, respectively) (P < 0.05); and the density of MMP-9 was found to be lower at S3 (133.60 pixels) than that at S1 and S2 (359.60 and 370.60 pixels, respectively). The density of proMMP-2 was low at S1 and S3 (130.45 and 145.20 pixels, respectively). On the other hand, the density of proMMP-9 was statistically different between S1 and S3 (89.22 vs. 415.60 pixels). Both proMMP-2 and proMMP-9 were absent at S2. Thus, MMP-2 and MMP9 exhibited opposite behaviors depending on the stage [...].(AU)


A encefalite por artrite caprina é uma doença lentiviral que leva a perdas consideráveis na criação de caprinos. Na fase aguda da infecção viral, embora os anticorpos antivirais sejam produzidos pelo sistema imunológico do hospedeiro, eles não são suficientes para serem detectados por testes sorológicos. As infecções agudas começam com um período de incubação, durante o qual o genoma viral se replica e as respostas inatas do hospedeiro são iniciadas. As metaloproteinases da matriz (MMPs) são enzimas que desempenham um papel importante nos processos fisiológicos e patológicos da remodelação tecidual. O presente estudo teve como objetivo avaliar a expressão de MMPs e sua atividade no soro sanguíneo de reprodutores caprinos infectados experimentalmente pelo vírus da encefalite por artrite caprina(CAEV). Cinco machos caprinos, com idades entre 3-4 anos, foram inoculadas por via intravenosa com a cepa de CAEV Cork (título: 105-6 TCID50/mL) após serem testados negativamente para CAEV três vezes em intervalos consecutivos de 30 dias usando a análise de western blot e nested-PCR. O estudo incluiu três etapas: estágio S1 ou pré-infecção; S2 ou estágio de soroconversão, correspondente à ocorrência de primeira soroconversão; e S3 ou pós-soroconversão, correspondendo a 23 semanas após a soroconversão. A zimografia foi realizada para as amostras utilizando gel de gelatina (12,5%), que foram submetidos à eletroforese em 170V, 1A e 300W por 50-70 min. Verificou-se que a densidade de MMP-2era menor em S1 (1456,20 pixels) do que em S2 e S3 (1943,80 e 2104,40 pixels, respectivamente) (P <0,05); e a densidade de MMP-9 foi menor em S3 (133,60 pixels) do que em S1 e S2 (359,60 e 370,60 pixels, respectivamente). A densidade do proMMP-2 era baixa em S1 e S3 (130,45 e 145,20 pixels, respectivamente). Por outro lado, a densidade do proMMP-9 foi estatisticamente diferente entre S1 e S3(89,22 vs. 415,60 pixels). ProMMP-2 e proMMP-9 estavam ausentes no S2. Assim, MMP-2 e [...].(AU)


Assuntos
Animais , Masculino , Lentivirus Ovinos-Caprinos/patogenicidade , Infecções por Lentivirus/sangue , Infecções por Lentivirus/enzimologia , Infecções por Lentivirus/veterinária , Ruminantes/sangue , Ruminantes/virologia , Metaloproteases/sangue
9.
Semina Ci. agr. ; 41(2): 531-544, Mar.-Apr. 2020. tab
Artigo em Inglês | VETINDEX | ID: vti-26932

RESUMO

This study aimed to determine the prevalence of ovine brucellosis in the states of Rio Grande do Norte, Paraíba, and Sergipe. A seroepidemiological survey was carried out in 121 properties under 23 municipalities between 2011 and 2012. The 1,034 serological samples collected from the sheep were subjected to the agar gel immunodiffusion (AGID) test using kits produced by the Paraná Institute of Technology (TECPAR). Anti-Brucella ovis antibodies were observed in 5.37% (70/1,304) of the animals evaluated, and the difference in the incidence of brucellosis between the adults and young animals of both sexes, and between the males and females was statistically significant (p < 0.05). The prevalence of brucellosis in Rio Grande do Norte, Paraíba, and was Sergipe 7.66% (36/470), 5.40% (13/241) and 3.54% (21/593), respectively. Of the municipalities visited, 91.30% (21/23) harbored herds with positive serology, and 42.15% (51/121) of the properties had at least one seropositive animal. The lack of training among the workers (p ? 0.05) (odds ratio = 3.68; 95% CI = 1.08-12.60; p = 0.038) was identified as a risk factor. The results obtained in this study indicate that the sheep from Rio Grande do Norte, Paraíba, and Sergipe are infected by B. ovis, thus necessitating the adoption of proper sanitary measures for preventing the spread of the disease in the herds.(AU)


O estudo teve como objetivo determinar a prevalência da Brucelose Ovina nos Estados do Rio Grandedo Norte, Paraíba e Sergipe. Foi realizado o levantamento soroepidemiológico em 121 propriedadesde 23 municípios durante os anos de 2011 e 2012. As 1.034 amostras sorológicas de ovinos foramsubmetidas à técnica de Imunodifusão em Gel de Ágar (IDGA), utilizando o kit produzido pelo Institutode Tecnologia do Paraná (TECPAR). Anticorpos anti-Brucella ovis foram observados em 5,37%(70/1.304) dos animais testados, havendo diferença estatística significativa (p < 0,05) para adultos ejovens de ambos os sexos, e entre machos e fêmeas. A prevalência encontrada no Estado do Rio Grandedo Norte, Paraíba e Sergipe foi de 7,66% (36/470), 5,40% (13/241) e 3,54% (21/593), respectivamente.Dos municípios visitados, 91,30% (21/23) eram compostos por rebanhos com sorologia positiva e42,15% (51/121) das propriedades possuíam pelo menos um animal soropositivo. A falta de capacitaçãode trabalhadores (p ≤ 0,05) (odds ratio = 3,68; IC 95% = 1,08-12,60; p = 0, 038) foi identificada comofator de risco. Os resultados obtidos neste trabalho indicam que a infecção por B. ovis encontra-sepresente nos ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe, sendo necessária a adoçãode medidas sanitárias para evitar a propagação da doença nos rebanhos.(AU)


Assuntos
Animais , Brucella ovis/patogenicidade , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/epidemiologia , Brucelose/epidemiologia , Brucelose/imunologia , Fatores de Risco
10.
Anim. Reprod. (Online) ; 16(4): 902-913, 2019. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1461477

RESUMO

The Saanen goat breed has been widely explored in breeding programmes; however, there are few reports about the breed’s genetic and molecular composition. Thus, this study aimed to characterize the proteomic profile of spermatozoa from Saanen breeding goats. Five breeding animals with proven fertility were selected, the spermatozoa were collected, and the protein was extracted. Subsequently, the proteins were separated and analysed by two-dimensional electrophoresis and mass spectrometry; the proteins were then identified with the SwissProt database. A total of 31 proteins involved in reproduction were identified, including binding proteins on spermatozoa for fusion with the egg, acrosomal membrane proteins, metabolic enzymes, heat shock proteins, cytoskeletal proteins and spermatozoa motility proteins. The characterization of such proteins clarifies the molecular mechanisms of spermatogenesis and the modifications that ensure the success of fertilization.


Assuntos
Masculino , Animais , Cabras/fisiologia , Espermatogênese , Espermatozoides/química , Proteômica/classificação
11.
Anim. Reprod. ; 16(4): 902-913, 2019. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-24191

RESUMO

The Saanen goat breed has been widely explored in breeding programmes; however, there are few reports about the breeds genetic and molecular composition. Thus, this study aimed to characterize the proteomic profile of spermatozoa from Saanen breeding goats. Five breeding animals with proven fertility were selected, the spermatozoa were collected, and the protein was extracted. Subsequently, the proteins were separated and analysed by two-dimensional electrophoresis and mass spectrometry; the proteins were then identified with the SwissProt database. A total of 31 proteins involved in reproduction were identified, including binding proteins on spermatozoa for fusion with the egg, acrosomal membrane proteins, metabolic enzymes, heat shock proteins, cytoskeletal proteins and spermatozoa motility proteins. The characterization of such proteins clarifies the molecular mechanisms of spermatogenesis and the modifications that ensure the success of fertilization.(AU)


Assuntos
Animais , Masculino , Cabras/fisiologia , Proteômica/classificação , Espermatozoides/química , Espermatogênese
12.
Acta sci. vet. (Impr.) ; 46: Pub.1592-2018. map, tab
Artigo em Português | VETINDEX | ID: biblio-1457882

RESUMO

Background: Chlamydophilosis is a disease caused by the bacterium Chlamydophila abortus, responsible for reproductive damage in small ruminants, and economic losses to animal production. In addition, it is classified as a zoonosis once it presents symptoms similar to numerous bacterial and viral infections of the reproductive system, making clinical diagnosis difficult. The objective of this work was to estimate the seroprevalence of anti-C. abortus antibodies in goats and to identify the risk factors for the disease in two mesoregions of Rio Grande do Norte, Brazil.Material, Methods & Results: This research was carried out in two mesoregions of the State of Rio Grande do Norte (Central Potiguar - CP and West Potiguar - WP) located in the Northeast of Brazil. These mesoregions together represent 83% of the regional herd. A total of 54 properties were studied. Those are distributed in seven municipalities: Afonso Bezerra, Angicos, Lages and Pedro Avelino, in the CP mesoregion; and Apodi, Caraúbas, and Mossoró, in the OP mesoregion. We used 540 animals from herds characterized as non-defined breed (NDB) and native breed, regardless of their productive functions, being frequent the consortium of caprine and ovine species. Blood samples were collected through jugular venipuncture by vacuum system with 5 mL tubes without anticoagulant. After collection, the tubes were centrifuged at 1500 g for 15 min. Detection of anti-Chlamydophila sp. antibodies was performed by the Complement Fixation Test. In parallel with the collection, a questionnaire was applied addressing general property and producer characteristics. Seroprevalence of chlamydophilosis in the CP mesoregion was 3.04% (7/230) for the animals and 26.1% (6/23) for the properties, whereas, in the WP mesoregion, the values were 4.2% (13/310) and 35.5% (11/31), respectively.[...]


Assuntos
Animais , Infecções por Chlamydophila/diagnóstico , Infecções por Chlamydophila/epidemiologia , Infecções por Chlamydophila/veterinária , Ruminantes/microbiologia , Brasil , Estudos Soroepidemiológicos , Fatores de Risco
13.
Acta sci. vet. (Online) ; 46: Pub. 1592, 2018. mapas, tab
Artigo em Português | VETINDEX | ID: vti-19254

RESUMO

Background: Chlamydophilosis is a disease caused by the bacterium Chlamydophila abortus, responsible for reproductive damage in small ruminants, and economic losses to animal production. In addition, it is classified as a zoonosis once it presents symptoms similar to numerous bacterial and viral infections of the reproductive system, making clinical diagnosis difficult. The objective of this work was to estimate the seroprevalence of anti-C. abortus antibodies in goats and to identify the risk factors for the disease in two mesoregions of Rio Grande do Norte, Brazil.Material, Methods & Results: This research was carried out in two mesoregions of the State of Rio Grande do Norte (Central Potiguar - CP and West Potiguar - WP) located in the Northeast of Brazil. These mesoregions together represent 83% of the regional herd. A total of 54 properties were studied. Those are distributed in seven municipalities: Afonso Bezerra, Angicos, Lages and Pedro Avelino, in the CP mesoregion; and Apodi, Caraúbas, and Mossoró, in the OP mesoregion. We used 540 animals from herds characterized as non-defined breed (NDB) and native breed, regardless of their productive functions, being frequent the consortium of caprine and ovine species. Blood samples were collected through jugular venipuncture by vacuum system with 5 mL tubes without anticoagulant. After collection, the tubes were centrifuged at 1500 g for 15 min. Detection of anti-Chlamydophila sp. antibodies was performed by the Complement Fixation Test. In parallel with the collection, a questionnaire was applied addressing general property and producer characteristics. Seroprevalence of chlamydophilosis in the CP mesoregion was 3.04% (7/230) for the animals and 26.1% (6/23) for the properties, whereas, in the WP mesoregion, the values were 4.2% (13/310) and 35.5% (11/31), respectively.[...](AU)


Assuntos
Animais , Ruminantes/microbiologia , Infecções por Chlamydophila/diagnóstico , Infecções por Chlamydophila/epidemiologia , Infecções por Chlamydophila/veterinária , Estudos Soroepidemiológicos , Fatores de Risco , Brasil
14.
Anim. Reprod. (Online) ; 15(1): 84-92, Jan.-Mar. 2018. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1461342

RESUMO

This study aimed to develop protocols for the extraction of sperm proteins from Moxotó goats (Capra hircus) and to compare the resulting proteomic maps. The sperm proteins were isolated using an extraction buffer containing 7 M urea and 2 M thiourea, 20 mM DTT, and one of the following detergents: 1% or 4% CHAPS; 1% or 4% SDS; 1% or 4% Triton X-100; or a combination of CHAPS and SDS. The 1-DE and 2-DE profiles of the isolated proteins revealed that the various isolation methods were efficient. Qualitative and quantitative differences in the 1-DE and 2-DE profiles were observed. 2-DE maps indicated that the amount and diversity of proteins visualized depended on the detergent that was used. Furthermore, this work revealed that the combination of detergents increased the resolution of some spots and retained the characteristics of the individual detergents, depending on their concentrations.


Assuntos
Animais , Espermatozoides , Isolamento Reprodutivo , Ruminantes/embriologia , Proteômica
15.
Anim. Reprod. ; 15(1): 84-92, Jan.-Mar. 2018. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18637

RESUMO

This study aimed to develop protocols for the extraction of sperm proteins from Moxotó goats (Capra hircus) and to compare the resulting proteomic maps. The sperm proteins were isolated using an extraction buffer containing 7 M urea and 2 M thiourea, 20 mM DTT, and one of the following detergents: 1% or 4% CHAPS; 1% or 4% SDS; 1% or 4% Triton X-100; or a combination of CHAPS and SDS. The 1-DE and 2-DE profiles of the isolated proteins revealed that the various isolation methods were efficient. Qualitative and quantitative differences in the 1-DE and 2-DE profiles were observed. 2-DE maps indicated that the amount and diversity of proteins visualized depended on the detergent that was used. Furthermore, this work revealed that the combination of detergents increased the resolution of some spots and retained the characteristics of the individual detergents, depending on their concentrations.(AU)


Assuntos
Animais , Ruminantes/embriologia , Espermatozoides , Isolamento Reprodutivo , Proteômica
16.
Acta sci. vet. (Online) ; 43: Pub. 1258, Feb. 6, 2015. ilus, tab
Artigo em Português | VETINDEX | ID: vti-24345

RESUMO

Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...(AU)


Assuntos
Animais , Ruminantes/sangue , Ruminantes/virologia , Metaloproteinases da Matriz/análise , Metaloproteinases da Matriz/sangue , Vírus da Artrite-Encefalite Caprina , Zona Semiárida
17.
Acta sci. vet. (Impr.) ; 43: Pub.1258-2015. ilus, tab
Artigo em Português | VETINDEX | ID: biblio-1457349

RESUMO

Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...


Assuntos
Animais , Metaloproteinases da Matriz/análise , Metaloproteinases da Matriz/sangue , Ruminantes/sangue , Ruminantes/virologia , Vírus da Artrite-Encefalite Caprina , Zona Semiárida
18.
Braz. j. vet. res. anim. sci ; 36(5): 253-7, 1999. tab
Artigo em Português | LILACS | ID: lil-285615

RESUMO

Com a finalidade de se investigar o efeito no Sistema Nervoso Central (SNC) do opióide antagonista naloxone hidrocloride sobre a liberaçäo do hormônio luteinizante (LH) em ovelhas ovariectomizadas e hipoglicêmicas, utilizaram-se oito fêmeas mestiças oriundas das raças Mule e Suffolk, pesando 65,7 ñ 3,6 kg. Duas semanas antes do início dos trabalhos, os animais foram canulados bilateralmente nos ventrículos. Foram feitos dois tratamentos (TI- animais näo-estressados; TII- animais estressados), que foram subdivididos em três grupos (soluçäo salina, 1 mg e 2 mg de naloxone). Os animais foram distribuídos, aleatoriamente, dentro das parcelas e foram feitas repetiçöes com intervalo de uma semana, até que se alcançassem quatro observaçöes por tratamento. No TI näo se observou alteraçäo nas concentraçöes de ß-endorfinas e LH, enquanto no TII, apesar de os animais nao apresentarem alteraçöes nos níveis de beta-endorfinas após injeçäo intracerebroventricular (i.c.v.) de 1 mg de naloxone, observou-se diminuiçäo significativa (p<0,05) após injeçäo (i.c.v.) de 2 mg. No TII, as concentraçöes de LH aumentaram significativamente (p<0,05) após injeçäo (i.c.v.) de 1 e 2 mg de naloxone. Conclui-se, portanto, que mesmo na ausência dos esteróides gonadais, os opióides endógenos estao envolvidos no controle do LH em animais hipoglicêmicos


Assuntos
Adulto , Feminino , Animais , Hormônio Luteinizante , Naloxona , Entorpecentes , Receptores Opioides , Reprodução , Ovinos , Estresse Fisiológico
19.
Semina ciênc. agrar ; 41(2): 531-544, 2020. tab
Artigo em Inglês | VETINDEX | ID: biblio-1501763

RESUMO

This study aimed to determine the prevalence of ovine brucellosis in the states of Rio Grande do Norte, Paraíba, and Sergipe. A seroepidemiological survey was carried out in 121 properties under 23 municipalities between 2011 and 2012. The 1,034 serological samples collected from the sheep were subjected to the agar gel immunodiffusion (AGID) test using kits produced by the Paraná Institute of Technology (TECPAR). Anti-Brucella ovis antibodies were observed in 5.37% (70/1,304) of the animals evaluated, and the difference in the incidence of brucellosis between the adults and young animals of both sexes, and between the males and females was statistically significant (p < 0.05). The prevalence of brucellosis in Rio Grande do Norte, Paraíba, and was Sergipe 7.66% (36/470), 5.40% (13/241) and 3.54% (21/593), respectively. Of the municipalities visited, 91.30% (21/23) harbored herds with positive serology, and 42.15% (51/121) of the properties had at least one seropositive animal. The lack of training among the workers (p ? 0.05) (odds ratio = 3.68; 95% CI = 1.08-12.60; p = 0.038) was identified as a risk factor. The results obtained in this study indicate that the sheep from Rio Grande do Norte, Paraíba, and Sergipe are infected by B. ovis, thus necessitating the adoption of proper sanitary measures for preventing the spread of the disease in the herds.


O estudo teve como objetivo determinar a prevalência da Brucelose Ovina nos Estados do Rio Grandedo Norte, Paraíba e Sergipe. Foi realizado o levantamento soroepidemiológico em 121 propriedadesde 23 municípios durante os anos de 2011 e 2012. As 1.034 amostras sorológicas de ovinos foramsubmetidas à técnica de Imunodifusão em Gel de Ágar (IDGA), utilizando o kit produzido pelo Institutode Tecnologia do Paraná (TECPAR). Anticorpos anti-Brucella ovis foram observados em 5,37%(70/1.304) dos animais testados, havendo diferença estatística significativa (p < 0,05) para adultos ejovens de ambos os sexos, e entre machos e fêmeas. A prevalência encontrada no Estado do Rio Grandedo Norte, Paraíba e Sergipe foi de 7,66% (36/470), 5,40% (13/241) e 3,54% (21/593), respectivamente.Dos municípios visitados, 91,30% (21/23) eram compostos por rebanhos com sorologia positiva e42,15% (51/121) das propriedades possuíam pelo menos um animal soropositivo. A falta de capacitaçãode trabalhadores (p ≤ 0,05) (odds ratio = 3,68; IC 95% = 1,08-12,60; p = 0, 038) foi identificada comofator de risco. Os resultados obtidos neste trabalho indicam que a infecção por B. ovis encontra-sepresente nos ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe, sendo necessária a adoçãode medidas sanitárias para evitar a propagação da doença nos rebanhos.


Assuntos
Animais , Brucella ovis/patogenicidade , Brucelose/epidemiologia , Brucelose/imunologia , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/epidemiologia , Fatores de Risco
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