Effect of monochrome light with different wavelengths on biochemical parameters of hens

Artificial light, as one of the environmental factors, plays a significant role in regulating the synthesis and secretion of hormones related to the coordination of parameters of life, growth, immunity, and reproductive functions of hens. The article aims to study the influence of monochrome light with different wavelengths on the biochemical parameters of hens` blood serum. Four groups of "Hy-Line W-36" crossbred hens were formed. Hens of the 1st group were kept using monochrome light with different wavelength lamps with a wavelength of ~ 460 nm, the 2nd group ~ 600 nm, the 3rd group ~ 630 nm, and the 4th group ~ 650 nm. It was found that the use of light with different wavelengths for keeping hens in cages of multilevel batteries affects hen' biochemical parameters, according to the research results. It was established that when using light with a wavelength of ~ 630 and ~ 650 nm, the indicators of clinical biochemistry of hens' blood serum were within the normal physiological values. Whereas, with the use of light with a wavelength of ~ 600 nm, an increase in the level of glucose, creatinine, total protein, total bilirubin, and phosphorus, a decrease in the ratio of calcium to phosphorus, in the activity of alkaline phosphatase, aspartate aminotransferase, and lactate dehydrogenase, were observed in the hens' blood serum. The use of light with a wavelength of ~ 460 nm was accompanied by a further increase in the level of glucose, creatinine, total protein, urea, total bilirubin, phosphorus, the activity of alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, and gamma-glutamyltransferase, a decrease in the ratio of calcium and phosphorus.

Behavior and blood profile in Friesian-Holstein dairy cows in the special region of Yogyakarta, Indonesia

The background of this study was blood profile and cows' behavior that could be used as a tool to diagnose reproductive status in the dairy cows. This research was aimed to determine biochemical blood profile, progesterone level, and behaviors of Friesian Holstein crossbreed dairy cow in the tropical climates. Fifteen Friesian-Holstein crossbreeds cows, in early lactation, reared by local farmers were divided into two barn types: individual and groups of cows. Blood samples were collected from the jugular vein using a 22G needle on days 0, 14, and 28. For the determination of blood profile, blood was collected into Vacuntainer® blood tubes containing ethylenediaminetetraacetic acid (EDTA) to arrest glycolysis. The blood was transported to the laboratory with a coolbox. The concentration of progesterone in plasma was assayed byenzyme-linked immunosorbent assay (ELISA) using a commercial kit and the biochemical blood profile serum was measured by using microlab 300. The variables observed were glucose, cholesterol, urea, and non-pregnant progesterone. Data obtained was examined by T-test analysis. The result showed that the Temperature-Humidity Index were 81.3 and 79.6 respectively, there were no significant differences between two observation, in terms of total feeding time (3.06±0.48 and 2.91±0.22 hours/day), total water consumption (86.23±0.58 and 85.37±0.88 l/day), total standing (12.70±0.21 and 12.83±0.34 hours/day), total laying (11.30±0.71 and 11.16±0.62 hours/day). The blood profile and progesterone levels were lower than the normal range in dairy cows. Also, the result indicated that blood parameters were not significantly different (P > 0.05) between individual pen and group. The result indicated that the progesterone level was not significant in two models of management dairy cows. The conclusion of the research is the climate can affect behavior and the blood parameters. Blood profiles [...].(AU)

Behavior and blood profile in Friesian-Holstein dairy cows in the special region of Yogyakarta, Indonesia

The background of this study was blood profile and cows' behavior that could be used as a tool to diagnose reproductive status in the dairy cows. This research was aimed to determine biochemical blood profile, progesterone level, and behaviors of Friesian Holstein crossbreed dairy cow in the tropical climates. Fifteen Friesian-Holstein crossbreeds cows, in early lactation, reared by local farmers were divided into two barn types: individual and groups of cows. Blood samples were collected from the jugular vein using a 22G needle on days 0, 14, and 28. For the determination of blood profile, blood was collected into Vacuntainer® blood tubes containing ethylenediaminetetraacetic acid (EDTA) to arrest glycolysis. The blood was transported to the laboratory with a coolbox. The concentration of progesterone in plasma was assayed byenzyme-linked immunosorbent assay (ELISA) using a commercial kit and the biochemical blood profile serum was measured by using microlab 300. The variables observed were glucose, cholesterol, urea, and non-pregnant progesterone. Data obtained was examined by T-test analysis. The result showed that the Temperature-Humidity Index were 81.3 and 79.6 respectively, there were no significant differences between two observation, in terms of total feeding time (3.06±0.48 and 2.91±0.22 hours/day), total water consumption (86.23±0.58 and 85.37±0.88 l/day), total standing (12.70±0.21 and 12.83±0.34 hours/day), total laying (11.30±0.71 and 11.16±0.62 hours/day). The blood profile and progesterone levels were lower than the normal range in dairy cows. Also, the result indicated that blood parameters were not significantly different (P > 0.05) between individual pen and group. The result indicated that the progesterone level was not significant in two models of management dairy cows. The conclusion of the research is the climate can affect behavior and the blood parameters. Blood profiles [...].

Diagnosis of bovine brucellosis in bulls by seroagglutination and seminal plasma agglutination tests / Diagnóstico da brucelose bovina em reprodutores através da soroaglutinação e sêmen plasma aglutinação

Brucellosis is a zoonotic disease with common agent is Brucella abortus in bovine specie. The disease in bulls is usually asymptomatic and could have absence of anti-B. abortus antibody in serum. It was investigated of the occurrence of bovine brucellosis in bulls, which involved testing blood serum and seminal plasma from 177 bulls used for natural mating or as semen donors. Blood was collected from all the animals by venipuncture of the jugular vein to obtain the serum samples, while seminal plasma was obtained from the animals by electroejaculation. Samples were tested in a screening test (rose Bengal test). In the second stage, the samples testing positive in the RBT were subjected to the confirmatory test (2-mercaptoethanol). None of the animals tested positive when blood serum was used. However, 5.06% of the bulls were considered reagent positive when seminal plasma was tested. It also sought to determine whether there was an association between the kind of production and the destination of animals with positive results. The calculated odds ratio indicated that bulls of the kind of production dairy/both (beef and dairy) had a higher risk of brucellosis infection. It is recommended that seminal plasma agglutination and seroagglutination techniques are applied concomitantly.(AU)

Brucelose é uma zoonose que nos bovinos tem como principal agente a bactéria Brucella abortus. A doença nos touros é geralmente assintomática e pode haver ausência de anticorpos anti- B. abortus no soro sanguíneo. O presente estudo teve como objetivo principal avaliar a ocorrência da brucelose bovina em reprodutores para isso foram empregados testes oficiais tendo como material o soro sanguíneo e o plasma seminal. Foram utilizados 177 machos bovinos destinados à monta natural ou doação de sêmen. O soro sanguíneo foi obtido através de venopunção da jugular e o plasma seminal através da eletroejaculação. As amostras foram submetidas ao teste de triagem (Antígeno acidificado tamponado) e na segunda etapa, apenas as amostras positivas na triagem foram submetidas ao exame confirmatório (2-mercaptoetanol). Não houveram animais positivos quando utilizado o soro sanguíneo. Para plasma seminal houve 5,06% de touros reagentes. Avaliou-se também se sistema produtivo ou a destinação do animal possuíam associação com resultados positivos. O cálculo de odds ratio demonstrou que os reprodutores bovinos do sistema produtivo misto/leite tem maior risco de infecção para brucelose. Recomenda-se que a técnica de sêmen plasma aglutinação seja feita em conjunto com a soroaglutinação.(AU)

Metabolismo energético, proteico e mineral de ovelhas Santa Inês hígidas e com mastite subclínica / Energy, protein and mineral metabolism in Santa Inês ewes, both healthy and with subclinical mastitis

Este estudo teve por objetivo avaliar o metabolismo energético, proteico e mineral de ovelhas Santa Inês hígidas e com mastite subclínica acompanhadas durante o final da gestação e na lactação. Foram acompanhadas ovelhas submetidas ao mesmo sistema de criação semi-intensivo. Os animais foram avaliados conforme os momentos a seguir: 10 dias que precedeu o parto (dap) e 15 dias pós parto (dpp), 30 dpp, 60 dpp e 90 dpp. Os metabólitos sanguíneos foram avaliados a partir do momento que antecedeu ao parto e os metabólitos no soro lácteo nos momentos subsequentes. Após exame clínico e bacteriológico foi realizada a triagem das ovelhas acompanhadas neste estudo, sendo 12 hígidas e 18 com mastite subclínica. Durante a lactação, mantendo os mesmos critérios de triagem, foram selecionadas 11 glândulas mamárias sadias e 20 infectadas, das quais foi colhido o leite para obtenção do soro lácteo. Foram mensurados no soro sanguíneo os metabólitos do perfil energético (ácidos graxos não esterificados (AGNEs), β-hidroxibutirato (BHB), frutosamina, colesterol e triglicérides), do perfil proteico (proteína total, albumina, uréia e creatinina) e do perfil mineral (ferro, cobre, zinco, magnésio, cálcio total, cálcio ionizado, sódio e potássio). No soro lácteo foram mensurados os íons cálcio, sódio e potássio, bem como os AGNEs e o BHB. A bioquímica sanguínea revelou haver influência (P<0,05) do período do periparto e da lactação sobre as concentrações sanguíneas dos AGNEs, BHB, colesterol, albumina, uréia, cálcio ionizado e no soro lácteo sobre o íon potássio. As ovelhas portadoras de mastite subclínica apresentaram valores sanguíneos superiores (P<0,05) de colesterol, albumina e cobre e no soro lácteo teores superiores do íon sódio e dos AGNEs e inferiores do íon potássio. O bom escore corporal das ovelhas observado durante o estudo aliado aos achados bioquímicos permitiu concluir ter ocorrido maior requerimento energético no primeiro mês da lactação, porém não o suficiente para desencadear qualquer transtorno metabólico e o aparecimento de um quadro de cetonemia, sendo estas discretas alterações mais expressivas nas ovelhas com mastite subclínica.(AU)

The study aimed to evaluate the energy, protein and mineral metabolism in Santa Inês ewes, healthy and with subclinical mastitis, followed up during late gestation and lactation periods. Ewes subjected to the same semi-intensive nursing system were followed up. The animals were evaluated according to the following stages: 10 days before parturition (dbp) and 15 days postpartum (dpp), 30 dpp, 60 dpp, and 90 dpp. Blood metabolites were evaluated starting from the stage previous to parturition and whey metabolites were evaluated in the subsequent stages. A screening of the ewes followed up in this study (12 healthy and 18 with subclinical mastitis) was performed after a clinical and bacteriological examination. During lactation, maintaining the same screening criteria, 11 healthy and 20 infected mammary glands were selected; the milk for whey extraction was collected from these glands. Energy profile metabolites (non-esterified fatty acids [NEFAs], β-hydroxybutyrate [BHB], fructosamine, cholesterol and triglycerides), protein profile (total protein, albumin, urea and creatinine) and mineral profile (iron, copper, zinc, magnesium, total calcium, ionized calcium, sodium, and potassium) were measured in the blood serum. Calcium, sodium and potassium ions, as well as NEFAs and BHB were measured in the whey. Blood biochemistry revealed an influence (P<0.05) of the peripartum and lactation periods on the blood concentrations of NEFAs, BHB, cholesterol, albumin, urea, ionized calcium. An analysis of the whey also revealed an influence on the potassium ion. Ewes with subclinical mastitis showed higher (P<0.05) blood levels of cholesterol, albumin and copper; higher sodium ion concentrations and NEFAs, and lower potassium ion in whey. Good physical score of ewes observed during this study, combined with the biochemical findings, allowed us to conclude that there was a larger energy requirement in the first month of lactation; however, this requirement was not enough to trigger any metabolic disorder or the emergence of ketonemia, and these discrete changes were more apparent in ewes with subclinical mastitis.(AU)

PADRÕES BIOQUIMICOS SÉRICOS E HEMATOLÓGICOS EM BOVINOS MESTIÇOS CONFORME IDADE E SEXO

Este estudo objetivou estabelecer padrões bioquímicos séricos e hematológicos de bovinos mestiços conforme a idade e os sexos. Foram utilizados 2.741 animais, classificados dentro das seguintes categorias: 0 a 12 meses: 321 fêmeas e 192 machos; 13 a 24 meses: 723 fêmeas e 598 machos; acima de 24 meses: 527 fêmeas e 380 machos. O experimento foi realizado na propriedade rural Fazenda União, município de Fama, Sul de MG. Os animais selecionados apresentaram estados clínicos normais. Para os exames bioquímicos séricos e hematológicos, as coletas de sangue foram realizadas por venopunção da jugular externa e retirados 20 mL de sangue por animal, em tubos de coleta a vácuo, sendo 10 mL sem anticoagulante para exames bioquímicos séricos e 10 mL com coagulante para os exames hematológicos. No laboratório, foram avaliadas e analisadas as amostras, para a determinação sérica dos seguintes parâmetros bioquímicos: creatinina, ureia, fosfatase alcalina, globulina, albumina, proteínas totais, transaminases glutâmicas oxalacéticas; gama glutamil transferase e calculada a relação albumina/globulina. Para os exames hematológicos, foram avaliadas e analisadas as amostras para as determinações séricas de contagem de hemácias; hemoglobina (Hb); hematócrito (Ht); volume corpuscular médio (VCM); concentração de hemoglobina globular média (CHGM); hemoglobina corpuscular média (HCM); leucócitos (LEU); neutrófilos segmentados (NSEG); linfócitos (LIN); monócitos (MON); eosinófilos (EOS) e plaquetas (PLA). Os dados foram submetidos à análise de variância e comparados pelo teste Tukey a um nível 5% de probabilidade. As variáveis bioquímicas séricas, creatinina, globulina, PT determinaram resultados (p<0,05) em fêmeas e machos até 12 meses de idade e em animais acima de 12 meses de idade. A FA e a GGT determinaram resultados (p<0,05) em animais de até 12 meses de idade e acima de 12 meses de idade. TGO determinou resultado (p<0,05) em animais entre 13 e 23 meses de idade. A albumina e a relação A/G não apresentaram diferenças (P>005) para as variáveis estudadas. As variáveis hematológicas, Ht, HCM, NSEG, LIN e EOS determinaram resultados (p<0,05) em animais de até 12 meses de idade e acima de 12 meses de idade. MON determinou resultado (p<0,05) em fêmeas e machos até 12 meses de idade e em animais acima de 12 meses de idade. As Hemácias, Hb, CHGM, LEU e PLA não determinaram diferenças nos resultados (P>005) para as variáveis estudadas. Conclui-se que o trabalho estabeleceu padrões bioquímicos séricos e hematológicos em bovinos mestiços conforme idade e sexo.

This work (study) aimed to establish serum and hematological biochemical patterns of crossbred cattle according to age and sex. 2,741 animals were used, classified into the following categories: 0 to 12 months: 321 females and 192 males; 13 to 24 months: 723 females and 598 males; above 24 months: 527 females and 380 males. The experiment was carried out at Fazenda União rural estate, municipality of Fama, south of MG. The animals selected had normal clinical status. Serum and hematological biochemical examinations were performed by venipuncture of the external jugular vein and 20 mL of blood were collected per animal in vacuum collection tubes, 10 mL without anticoagulant for serum biochemical tests and 10 mL with coagulant for hematological examinations. In the laboratory, the following biochemical parameters were evaluated and analyzed: creatinine, urea, alkaline phosphatase, globulin, albumin, total proteins, glutamic oxalacetic transaminases; gamma glutamyl transferase and calculated the albumin / globulin ratio. For the hematological examinations, the samples were evaluated and analyzed for the serum determinations of red blood cell count; hemoglobin (Hb); hematocrit (Ht); mean corpuscular volume (MCV); mean globular hemoglobin concentration (CHGM); mean corpuscular hemoglobin (HCM); leukocytes (LEU); Segmented neutrophils (NSEG); lymphocytes (LIN); monocytes (MON); eosinophils (EOS) and platelets (PLA). Data were submitted to analysis of variance and compared by Tukey test at a 5% probability level. The serum biochemical variables, creatinine, globulin, PT determined results (p <0.05) in females and males up to 12 months of age and in animals over 12 months of age. AF and GGT determined results (p <0.05) in animals up to 12 months of age and above 12 months of age. TGO determined outcome (p <0.05) in animals between 13 and 23 months of age. Albumin and the A / G ratio did not present differences (P> 005) for the studied variables. Hematological variables, Ht, HCM, NSEG, LIN and EOS determined results (p <0.05) in animals up to 12 months of age and above 12 months of age. MON determined results (p <0.05) in females and males up to 12 months of age and in animals over 12 months of age. Haemocytes, Hb, CHGM, LEU and PLA did not determine differences in results (P> 005) for the variables studied. It is concluded that the work established serum and hematological biochemical patterns in crossbred cattle according to age and sex.

Lipid peroxidation in rats treated with vincristine sulphate and nandrolone decanoate / Peroxidação lipídica em ratos tratados com sulfato de vincristina e decanoato de nandrolona

Brain and serum lipid peroxidation was studied in rats treated with vincristine sulphate and different doses of nandrolone decanoate. Thirty rats were distributed into six groups (n=5). The treatments were applied once a week for two weeks. Sample collection was performed in the third week. Treatments during the first week were: G1 (control) - physiologic solution, G2 - vincristine sulphate (4mg/m²), G3 - physiologic solution, G4 - physiologic solution, G5- vincristine sulphate (4mg/m²), and G6 - vincristine sulphate (4mg/m²). In the second week, they were: G1 (control) - physiologic solution, G2- physiologic solution, G3 - nandrolone decanoate (1.8mg/kg-1), G4 - nandrolone decanoate (10mg/kg-1), G5 - nandrolone decanoate (1.8mg/kg-1), and G6 - nandrolone decanoate (10mg/kg-1). Lipid peroxidation increased with the isolated use of vincristine and nandrolone decanoate, and with vincristine associated to the highest dose of the ester as well. These results suggest that vincristine sulphate and nandrolone decanoate increase free radical production. Therapeutic dose of nandrolone decanoate when associated with vincristine sulphate proved to be beneficial, as it was able to protect the organism from damaging processes involved in free radical production.(AU)

Este estudo teve por objetivo detectar a peroxidação lipídica presente no cérebro e no soro de ratos tratados com sulfato de vincristina e diferentes doses de decanoato de nandrolona. Trinta ratos foram distribuídos em seis grupos (n=5). Os tratamentos foram aplicados uma vez por semana, durante duas semanas, e a coleta de amostras foi realizada na terceira semana. Na primeira semana, os tratamentos consistiram de: G1(controle) - solução fisiológica; G2 - sulfato de vincristina (4mg/m 2 ); G3 - solução fisiológica; G4 - solução fisiológica; G5 - sulfato de vincristina (4mg/m 2 ) e G6 - sulfato de vincristina (4mg/m 2 ). Na segunda semana: G1(controle) - solução fisiológica; G2 - solução fisiológica; G3 - decanoato de nandrolona (1.8mg/kg-1 ); G4 - decanoato de nandrolona (10mg/kg-1 ); G5 - decanoato de nandrolona (1.8mg/kg-1 ) e G6 - decanoato de nandrolona (10mg/kg-1 ). A peroxidação lipídica aumentou com o uso isolado tanto da vincristina quanto do decanoato de nandrolona e com a associação da vincristina à dose mais alta do éster. Estes resultados sugerem que o sulfato de vincristina e o decanoato de nandrolona aumentam a produção de radicais livres. A dose terapêutica do decanoato de nandrolona, quando associada ao sulfato de vincristina, provou ser benéfica, já que foi capaz de proteger o organismo dos processos prejudiciais induzidos pela produção de radicais livres.(AU)

Susceptibility of Arcobacter butzleri to human blood serum / Susceptibilidade de Arcobacter butzleri ao soro humano

A susceptibilidade de 10 amostras de Arcobacter butzleri ao soro humano foi estudada. A maior atividade bactericida foi encontrada no soro humano normal, com taxas de sobrevivência bacterianainversamente proporcionais à diluição do soro. As maiores taxas de sobrevivência foram obtidas com o soro inativado pelo calor. As taxas de sobrevivência decresceram com a adição de soro fresco ao inativado. O soro com valores reduzidos de gamaglobulinas e valores normais de complemento mostrou ativo efeito bactericida. Os resultados demonstraram que A. butzleri é altamente susceptível ao efeito bactericida do soro humano, sugerindo que pode ser capaz de ativar diretamente o complemento pela via alternativa.(AU)

Blood serum levels of calcium phosphorus and magnesium during the pregnancy of the she ass, Equus asinus, L. 1758 (Perissodactyla, Equidae) / Contribuição ao estudo dos teores séricos sanguíneos do cálcio, fósforo e magnésio durante a prenhez da jumenta, Equus asinus, Linnaeus, 1758. (Perissodactyla, Equidae)

In this work the blood serum levels of calcium, inorganic phosphorus and magnesium during pregnancy of the she ass have been carried out. Samples of blood have been collected from 15 she asses in differents periods of the pregnancy and in the post-partum. Total calcium and magnesium were determined by the TODD method and the inorganic phosphorus by the FISKE-SUBBAROW method. The levels of the total calcium were higher in the final periodo of pregnancy as well as in post-partum, with a statistical significance at the level of 1%. The results of the inorganic phosphorus and magnesium have not shown statistical significance differences among them.

Foram analisados os níveis séricos sanguíneos do cálcio, fósforo e magnésio durante a prenhez da jumenta. As amostras foram colhidas de 15 jumentas, em diferentes períodos da gestação e no pós-parto. A dosagem do cálcio e magnésio foi efetuada pelo método de TODD e a do fósforo pela técnica de FISKE-SUBBAROW. Para o cálcio verificou-se que na jumenta prenhe, a calcemia tende a se elevar com o progredir da gestação, atingindo teores maiores no pré-parto e assim se mantendo até o 3. dia do puerpério, com uma significância estatística ao nível de 1%. Com relação aos teores do fósforo inorgânico e do magnésio, não se obteve significância estatística dos resultados.

Blood serum levels of calcium phosphorus and magnesium during the pregnancy of the she ass, Equus asinus, L. 1758 (Perissodactyla, Equidae) / Contribuição ao estudo dos teores séricos sanguíneos do cálcio, fósforo e magnésio durante a prenhez da jumenta, Equus asinus, Linnaeus, 1758. (Perissodactyla, Equidae)

In this work the blood serum levels of calcium, inorganic phosphorus and magnesium during pregnancy of the she ass have been carried out. Samples of blood have been collected from 15 she asses in differents periods of the pregnancy and in the post-partum. Total calcium and magnesium were determined by the TODD method and the inorganic phosphorus by the FISKE-SUBBAROW method. The levels of the total calcium were higher in the final periodo of pregnancy as well as in post-partum, with a statistical significance at the level of 1%. The results of the inorganic phosphorus and magnesium have not shown statistical significance differences among them.

Foram analisados os níveis séricos sanguíneos do cálcio, fósforo e magnésio durante a prenhez da jumenta. As amostras foram colhidas de 15 jumentas, em diferentes períodos da gestação e no pós-parto. A dosagem do cálcio e magnésio foi efetuada pelo método de TODD e a do fósforo pela técnica de FISKE-SUBBAROW. Para o cálcio verificou-se que na jumenta prenhe, a calcemia tende a se elevar com o progredir da gestação, atingindo teores maiores no pré-parto e assim se mantendo até o 3. dia do puerpério, com uma significância estatística ao nível de 1%. Com relação aos teores do fósforo inorgânico e do magnésio, não se obteve significância estatística dos resultados.

Diagnosis of bovine brucellosis in bulls by seroagglutination and seminal plasma agglutination tests / Diagnóstico da brucelose bovina em reprodutores através da soroaglutinação e sêmen plasma aglutinação

Brucellosis is a zoonotic disease with common agent is Brucella abortus in bovine specie. The disease in bulls is usually asymptomatic and could have absence of anti-B. abortus antibody in serum. It was investigated of the occurrence of bovine brucellosis in bulls, which involved testing blood serum and seminal plasma from 177 bulls used for natural mating or as semen donors. Blood was collected from all the animals by venipuncture of the jugular vein to obtain the serum samples, while seminal plasma was obtained from the animals by electroejaculation. Samples were tested in a screening test (rose Bengal test). In the second stage, the samples testing positive in the RBT were subjected to the confirmatory test (2-mercaptoethanol). None of the animals tested positive when blood serum was used. However, 5.06% of the bulls were considered reagent positive when seminal plasma was tested. It also sought to determine whether there was an association between the kind of production and the destination of animals with positive results. The calculated odds ratio indicated that bulls of the kind of production dairy/both (beef and dairy) had a higher risk of brucellosis infection. It is recommended that seminal plasma agglutination and seroagglutination techniques are applied concomitantly.

Brucelose é uma zoonose que nos bovinos tem como principal agente a bactéria Brucella abortus. A doença nos touros é geralmente assintomática e pode haver ausência de anticorpos anti- B. abortus no soro sanguíneo. O presente estudo teve como objetivo principal avaliar a ocorrência da brucelose bovina em reprodutores para isso foram empregados testes oficiais tendo como material o soro sanguíneo e o plasma seminal. Foram utilizados 177 machos bovinos destinados à monta natural ou doação de sêmen. O soro sanguíneo foi obtido através de venopunção da jugular e o plasma seminal através da eletroejaculação. As amostras foram submetidas ao teste de triagem (Antígeno acidificado tamponado) e na segunda etapa, apenas as amostras positivas na triagem foram submetidas ao exame confirmatório (2-mercaptoetanol). Não houveram animais positivos quando utilizado o soro sanguíneo. Para plasma seminal houve 5,06% de touros reagentes. Avaliou-se também se sistema produtivo ou a destinação do animal possuíam associação com resultados positivos. O cálculo de odds ratio demonstrou que os reprodutores bovinos do sistema produtivo misto/leite tem maior risco de infecção para brucelose. Recomenda-se que a técnica de sêmen plasma aglutinação seja feita em conjunto com a soroaglutinação.

Comparison of different protocols for the bovine viral diarrhea virus detection by RT-PCR in pools of whole blood and blood serum artificially contaminated / Comparação de diferentes protocolos para a detecção do vírus da diarréia viral bovina por RT-PCR em grupos de sangue total e de soro sangüíneo, artificialmente contaminados

The RT-PCR technique was optimized and evaluated to detect the 5 untranslated region of bovine viral diarrhea virus (BVDV) from clinical samples that consisted of blood serum and whole blood artificially contaminated with the NADL strain of BVDV. To optimization of technique, the following conditions were evaluated: i) two pairs of primers, 103 / 372 (WEINSTOCK et al., 2001) and 324 / 326 (VILCEK et al., 1994), ii) four methods of nucleic acid extraction (phenol/chloroform/isoamyl alcohol; silica/guanidine isothiocyanate; a combination of the two previous methods; and TRIzol) and iii) different concentrations and compositions of reagents and time/temperature of the reactions. Between the alternatives tested that resulted in the amplification of the 290 bp product that was easily visualized in ethidium bromide stained 2% agarose gel was that presented the following conditions: i) primers 103 and 372; ii) initial volume and clinical sample: 50 mL of blood serum; iii) extraction of nucleic acid: silica/guanidine isothiocyanate method; iv) reverse transcription: 9 mL extracted nucleic acid, 1xPCR buffer (20 mM Tris-HCl pH 8.4 and 50 mM KCl), 1.5 mM MgCl2; 60 units of reverse transcriptase enzyme M-MLV, RNA denaturation at 97C / 4 min, and reverse transcription at 42C / 30 min; v) PCR: primers 103 / 372 with anneling temperature at 59C. The utilization of RT-PCR within these op

A técnica da RT-PCR foi otimizada e avaliada para a detecção da região 5 terminal não-traduzida do genoma do vírus da diarréia viral bovina (BVDV) em amostras clínicas de bovinos, constituídas por soro sangüíneo e sangue total, artificialmente contaminadas com a estirpe NADL do BVDV. Para a otimização da técnica foram avaliados: i) dois pares de primers, 103 / 372 (WEINSTOCK et al., 2001) e 324 / 326 (VILCEK et al., 1994) ii) quatro métodos de extração do ácido nucléico (fenol / clorofórmio / álcool isoamílico; sílica / isotiocianato de guanidina; uma combinação dos dois métodos anteriores e TRIzol) e iii) diferentes concentrações e composições de reagentes, tampões e tempo/temperatura das reações. Entre todas as alternativas testadas a que resultou na amplificação de um produto com 290 pb, que foi facilmente visualizado em gel de agarose 2% com brometo de etídeo, foi a que empregou as seguintes condições: i) primers 103 e 372; ii) volume inicial e amostra clínica: 50 mL de soro sangüíneo; iii) extração do ácido nucléico: método da sílica / isotiocianato de guanidina, iv) transcrição reversa: 9 mL do ácido nucléico extraído, 1xPCR buffer (20 mM Tris-HCl pH 8,4 e 50 mM KCl), 1,5 mM MgCl2, 60 unidades da enzima transcriptase reversa M-MLV, desnaturação do RNA a 97C / 4 min e transcrição reversa a 42C / 30 min e v) PCR: primers 103 / 372 com temperatura de anelamento de 59C.