PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Silva W.B., Machado C., Goldberg D.W., Moreira S.B., Silva L.C.C.P., Freire I.M.A., Maciel P.O. & Almosny N.R. 2009. [Assessment of blood inflammatory response in BCG stimulated rattlesnakes (Crotalus durissus Linnaeus, 1758).] Avaliação da resposta hematológica em cascavéis (Crotalus durissus Linnaeus, 1758) inoculadas com BCG. Pesquisa Veterinária Brasileira 29(12):985-992. Programa de Pós-Graduação em Clínica e Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Federal Fluminense, Rua Vital Brazil Filho 64, Niterói, RJ 24230-360, Brazil. E-mail: wellban@yahoo.com.br The high demand for anti-venom production in response to the increased number of cases of snakebite envenomation highlights the importance of raising and breeding venomous snakes in captivity. Knowledge of types of venoms and anti-venoms is of great interest to public health. The maintenance of venomous serpents in captivity started in the early twentieth century, but still nowadays it is a challenge to manage and prevent diseases in captive fauna. Hematology is commonly used for general health assessment and illness detection. However, data on serpent blood analysis are still scarce. Alterations in hematological parameters were experimentally induced in rattlesnakes by the inoculation of BCG. Based on this, hemograms can be used as a health auxiliary diagnosis method for bacterial diseases in this species. In this study, blood samples were taken from 10 healthy specimens of rattlesnakes (Crotalus durissus) born and bred in captivity in the Herpetological Division of Vital Brazil Institute. Animals were divided into two groups (group 1 and 2) with similar live weight and sex proportion, and were then inoculated subcutaneously with BCG (Bacillus Calmette-Guérin). Blood samples were taken before and after inoculation at three experimental times (days 3, 5 and 7 for group 1 and days 11, 17 and 21 for group 2). Hematological analysis was performed through semi-direct technique, blood samples were diluted in Natt and Herrick solution and smears were stained by Giemsa. Serpents from group 1 developed discrete anemia due to the inflammatory syndrome, and showed significant decrease of MCH and MCHC. Granulocytes were characterized by the presence of rough granules. The azurophils varied in shape and size and showed large amount of cytoplasmic vacuoles. The thrombocytopenia observed in group 2 suggests that these cells participate in the inflammatory process. A single individual from group 1 showed granulocytosis and a few animals showed a discrete azurophilia. In general, BCG inoculation unleashes hematological inflammatory responses characterized by the presence of thrombocytes, azurophils and granulocytes cells.

• Cascavel Crotalus durissus

Abstract in Portuguese:

RESUMO.- Silva W.B., Machado C., Goldberg D.W., Moreira S.B., Silva L.C.C.P., Freire I.M.A., Maciel P.O. & Almosny N.R. 2009. [Assessment of blood inflammatory response in BCG stimulated rattlesnakes (Crotalus durissus Linnaeus, 1758).] Avaliação da resposta hematológica em cascavéis (Crotalus durissus Linnaeus, 1758) inoculadas com BCG. Pesquisa Veterinária Brasileira 29(12):985-992. Programa de Pós-Graduação em Clínica e Reprodução Animal, Faculdade de Medicina Veterinária, Universidade Federal Fluminense, Rua Vital Brazil Filho 64, Niterói, RJ 24230-360, Brazil. E-mail: wellban@yahoo.com.br A criação de serpentes peçonhentas em cativeiro para produção de soros antipeçonhas possui crescente importância para a saúde pública devido ao aumento do número de notificações de acidentes ofídicos a cada ano no Brasil. Iniciado no século XX, ainda hoje essa atividade apresenta alguns desafios como a instalação de doenças no plantel. O hemograma é um exame de triagem clínica que auxilia no diagnóstico de diversas moléstias que acometem diferentes espécies de animais, no entanto ainda pouco estudado em serpentes. A caracterização das alterações hematológicas em cascavéis inoculadas experimentalmente com BCG pode servir de base na utilização deste exame no auxílio ao diagnóstico de infecções bacterianas na espécie. Dessa forma, foram realizados exames hematológicos em 10 serpentes da espécie Crotalus durissus pertencentes ao plantel da Divisão de Herpetologia do Instituto Vital Brazil. Os animais foram divididos em dois grupos (Grupos 1 e 2), homogêneos entre si em relação ao peso e proporção sexual. Os dois grupos foram inoculados com BCG e submetidos à coleta de sangue antes da inoculação e em três momentos pós-inoculação (3º, 5º, e 7º dias para o Grupo 1 e 11º, 17º e 21º dias para o Grupo 2). O hemograma foi realizado por método semidireto pela utilização de líquido de Natt e Herrick e as lâminas foram coradas pelo Giemsa. Observou-se anemia discreta, com redução dos valores de concentração de hemoglobina corpuscular média e da hemoglobina globular média no Grupo 1 que foi relacionada à doença inflamatória. A trombocitopenia observada no Grupo 2 sugeriu a atuação deste tipo celular em processos inflamatórios. Um único animal do Grupo 1 apresentou granulocitose e alguns animais apresentaram discreta azurofilia. Observaram-se alterações morfológicas nos leucócitos. Os granulócitos apresentaram granulações grosseiras e os azurófilos apresentaram aumento de tamanho e grandes vacúolos. De forma geral, a inoculação de BCG em cascavéis desencadeia respostas inflamatórias hematológicas caracterizadas pela participação de trombócitos, granulócitos e azurófilos.

Abstract in English:

ABSTRACT.- Silverio S.M., Mari R.B., Clebis N.K., Scoz J.R., Germano R.M., Agreste F., Bombonato P.P. & Stabille S.R. 2008. Assessment of NADPH-diaphorase stained myenteric neurons of the jejunum of diabetic rats supplemented with ascorbic acid. Pesquisa Veterinária Brasileira 28(2):95-102. Departamento de Ciências Biológicas, UNIPAR, Campus-Paranavaí, Av. Huberto Brüning 360, Jardim Santos Dumont, Paranavaí, PR 87706-490, Brazil. E-mail: srstabille@wnet.com.br The relation between hyperglycemia and diabetic neuropathy has already been demonstrated in some studies. Among the theories proposed for its etiology the oxidative stress stands out. The performance of nitric oxide as a link between the metabolic and vascular neuropathogenic factors that triggers the diabetic neuropathy has already been put forward. This study aimed to assess the quantification and measurements of the cell body profile area (CBPA) of NADPH-diaphorase reactive (NADPH-dp) myenteric neurons of the jejunum of diabetic rats (induced by streptozotocin) supplemented with Ascorbic Acid (AA). These changes in the myenteric neurons seem to be related to the gastrointestinal disturbances observed in diabetes mellitus (DM). Twenty male Wistar rats (Rattus norvegicus) were distributed in 4 groups (n=5): controls (C), control supplemented (CS), diabetic (D), and diabetic suplemented (DS). DM was induced by estreptozotocin (50mg/kg body wt). One week after the induction and confirmation of the DM (glycemia exam), animals of the groups CS and DS received 50mg of AA three times a week by gavage. After 90 days of experiment, the animals were anesthetized with lethal thiopental dose (40mg/kg) and the collected jejunum processed for the histochemistry NADPH-diaphorase technique. Whole-mount preparations were obtained for quantitative and morphometric analysis of the myenteric neurons. A quantity of jejunum neurons in the Group D (96±7.5) was not different (P>0.05) from Group DS (116±8.08), C (92±9.7), and CS (81±5.4), but in Group DS the quantity was higher (P<0.05) than in Group C and CS. The CBPA of neurons from Group D (189.50±2.68µm2) and DS (195.92±3.75µm2) were lower (P<0.05) than from Group C (225.13±4.37µm2) and CS (210.23±3.15µm2). The streptozotocin-induced DM did not change the jejunum-ileum area, the jejunum myenteric plexus space organization and the density of NADPH-dp neurons. The 50g AA-supplementation, three times a week, during 90 days, did not decrease hyperglycemia; however, it had a neuroprotective effect on the myenteric neurons, minimizing the increase on the CBPA of NADPH-dp neurons and increasing the amount of NADPD-dp neurons.

• Myenteric plexus intestine vitamin C antioxidants hyperglycemia

Abstract in Portuguese:

ABSTRACT.- Silverio S.M., Mari R.B., Clebis N.K., Scoz J.R., Germano R.M., Agreste F., Bombonato P.P. & Stabille S.R. 2008. Assessment of NADPH-diaphorase stained myenteric neurons of the jejunum of diabetic rats supplemented with ascorbic acid. Pesquisa Veterinária Brasileira 28(2):95-102. Departamento de Ciências Biológicas, UNIPAR, Campus-Paranavaí, Av. Huberto Brüning 360, Jardim Santos Dumont, Paranavaí, PR 87706-490, Brazil. E-mail: srstabille@wnet.com.br The relation between hyperglycemia and diabetic neuropathy has already been demonstrated in some studies. Among the theories proposed for its etiology the oxidative stress stands out. The performance of nitric oxide as a link between the metabolic and vascular neuropathogenic factors that triggers the diabetic neuropathy has already been put forward. This study aimed to assess the quantification and measurements of the cell body profile area (CBPA) of NADPH-diaphorase reactive (NADPH-dp) myenteric neurons of the jejunum of diabetic rats (induced by streptozotocin) supplemented with Ascorbic Acid (AA). These changes in the myenteric neurons seem to be related to the gastrointestinal disturbances observed in diabetes mellitus (DM). Twenty male Wistar rats (Rattus norvegicus) were distributed in 4 groups (n=5): controls (C), control supplemented (CS), diabetic (D), and diabetic suplemented (DS). DM was induced by estreptozotocin (50mg/kg body wt). One week after the induction and confirmation of the DM (glycemia exam), animals of the groups CS and DS received 50mg of AA three times a week by gavage. After 90 days of experiment, the animals were anesthetized with lethal thiopental dose (40mg/kg) and the collected jejunum processed for the histochemistry NADPH-diaphorase technique. Whole-mount preparations were obtained for quantitative and morphometric analysis of the myenteric neurons. A quantity of jejunum neurons in the Group D (96±7.5) was not different (P>0.05) from Group DS (116±8.08), C (92±9.7), and CS (81±5.4), but in Group DS the quantity was higher (P<0.05) than in Group C and CS. The CBPA of neurons from Group D (189.50±2.68µm2) and DS (195.92±3.75µm2) were lower (P<0.05) than from Group C (225.13±4.37µm2) and CS (210.23±3.15µm2). The streptozotocin-induced DM did not change the jejunum-ileum area, the jejunum myenteric plexus space organization and the density of NADPH-dp neurons. The 50g AA-supplementation, three times a week, during 90 days, did not decrease hyperglycemia; however, it had a neuroprotective effect on the myenteric neurons, minimizing the increase on the CBPA of NADPH-dp neurons and increasing the amount of NADPD-dp neurons.

Abstract in English:

Thirty one farms, located throughout the county of Bagé, Rio Grande do Sul, were assessed for anthelmintic usage and for sheep nematode resistance to anthelmintics. On each farm, 30 weaned lambs were weighed, ear-tagged and divided into three groups of ten. One group received thiabendazole at 66.75mg/kg, the second tetramisole at 12.6mg/kg, while the third remained untreated as a control. All lambs were drenched according to their individual body weight. Faeces were collected from all animals on the day of treatment and 7 days later to provide material for egg counts and larval cultures. Between the 31 surveyed farms, 38.7% showed benzimidazole resistance, 25.8% resistance to leva/tetramisole and 19.4% had multiple resistance. At the time of the assessment all farmers were using broad spectrum anthelmintics. The results of larval cultures showed that larvae surviving the treatment with thiabendazole were mainly Haemonchus, while those surviving tetramisole treatment were Trichostrongylus spp and Ostertagia spp.

• Anthelmintic resistance sheep assessment

Abstract in Portuguese:

A avaliação de resistência de parasitos aos anti-helmínticos a nível de campo foi realizado em 31 rebanhos de ovinos de Bagé, RS. Em cada fazenda foram separados 30 cordeiros desmamados (nascidos no inverno-primavera anterior) os quais foram individualmente pesados, identificados e distribuídos em três tratamentos: 1) Thiabendazole = 66,75mg/kg; 2) Tetramisole = 12,6mg/kg e 3) Controle = não medicado. Os anti-helmínticos foram administrados conforme o peso individual de cada cordeiro. Fezes foram coletadas para OPG e cultura de larvas no dia da medicação e 7 dias após. Das 31 propriedades estudadas 38,7% apresentaram resistência aos benzimidazóis, 25,8% aos leva/tetramisóis e 19,4% resistência múltipla. Apenas 16,1 % apresentavam-se totalmente sensíveis aos anti-helnúnticos. Os resultados das culturas de larvas revelaram que as larvas que sobreviveram a medicação com thiabendazole eram principalmente Haemonchus spp enquanto que as sobreviventes ao tratamento com leva/tetramisole eram principalmente Trichostrongylus spp e Ostertagia spp.

• Resistência anti-helmíntica ovinos avaliação