PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Vasconcellos A.S., Chelini M.O.M., Palme R., Guimarães M.A.B.V., Oliveira C.A. & Ades C. 2011. Comparison of two methods for glucocorticoid evaluation in maned wolves. Pesquisa Veterinária Brasileira 31(Supl.1):79-83. Department of Experimental Psychology, Institute of Psychology, University of São Paulo, Av. Professor Mello de Moraes 1721, São Paulo, SP 05508-030, Brazil. E-mail: angelv@usp.br Analysis of faecal glucocorticoid metabolites provides a powerful noninvasive tool for monitoring adrenocortical activity in wild animals. However, differences regarding the metabolism and excretion of these substances make a validation for each species and sex investigated obligatory. Although maned wolves (Chrysocyon brachyurus) are the biggest canids in South America, their behaviour and physiology are poorly known and they are at risk in the wild. Two methods for measuring glucocorticoid metabolites in maned wolves were validated: a radio- and an enzyme immunoassay. An ACTH challenge was used to demonstrate that changes in adrenal function are reflected in faecal glucocorticoid metabolites. Our results suggest that both methods enable a reliable assessment of stress hormones in maned wolves avoiding short-term rises in glucocorticoid concentrations due to handling and restraint. These methods can be used as a valuable tool in studies of stress and conservation in this wild species.

• ACTH corticosterone cortisol corticosterona

Abstract in Portuguese:

RESUMO.- Vasconcellos A.S., Chelini M.O.M., Palme R., Guimarães M.A.B.V., Oliveira C.A. & Ades C. 2011. Comparison of two methods for glucocorticoid evaluation in maned wolves. [Comparação de dois métodos para avaliação de glicocorticóides em lobos guarás.] Pesquisa Veterinária Brasileira 31(Supl.1):79-83. Department of Experimental Psychology, Institute of Psychology, University of São Paulo, Av. Professor Mello de Moraes 1721, São Paulo, SP 05508-030, Brazil. E-mail: angelv@usp.br A análise de metabólitos de glicocorticóides fecais é uma ferramenta não-invasiva poderosa para o monitoramento da atividade adrenal em animais silvestres. Entretanto, diferenças no metabolismo e na excreção dessas substâncias tornam obrigatória a validação desse método para cada espécie e sexo investigado. Embora os lobos guarás (Chrysocyon brachyurus) sejam os maiores canídeos da América do Sul, seu comportamento e fisiologia são ainda pouco conhecidos e estes animais são ameaçados. Dois métodos para mensuração de metabólitos de glicocorticóides em lobos guarás foram validados: um radio- e um enzima- imuno-ensaio. Um desafio de ACTH foi usado para demonstrar que alterações no funcionamento adrenal podem ser detectadas nas concentrações metabólitos de glicocorticóides fecais. Nossos resultados sugerem que ambos os métodos possibilitam uma avaliação confiável dos hormônios relacionados ao estresse em lobos guarás, evitando assim aumentos de curta duração nas concentrações glicocorticóides, devidos ao manejo e contenção. Esses métodos podem ser usados como uma ferramenta valiosa em estudos de estresse e conservação com essa espécie silvestre.

Abstract in English:

ABSTRACT.- Barbosa J.D., Sousa M.G.S., Tokarnia C.H., Brito M.F., Belo Reis A.S., Bomjardim H.A., Lopes C.T.A. & Oliveira C.M.C. 2011. [Comparison between the clinical and pathological picture in the experimental poisoning by crotalic venom.] Quadro clínico-patológico do envenenamento crotálico experimental em bubalinos comparado com o de bovinos. Pesquisa Veterinária Brasileira 31(11):967-973. Central de Diagnóstico Veterinário, Universidade Federal do Pará, Rua Maximino Porpino da Silva 1000, Pirapora, Castanhal, PA 68743-080, Brazil. E-mail: diomedes@ufpa.br The objective of the study was to verify the sensibility of buffaloes to the poison of Crotalus durissus terrificus and to study the clinical-pathological picture in buffaloes in comparison with the one in cattle. The subcutaneous inoculation of the liofilized poison of the snake, diluted in 1ml of physiologic solution, was done in the area of the humerus-radio-ulnar joint of three buffaloes at doses of 0.015, 0.03 and 0.066mg/kg, and of two cattle at doses of 0.03 and 0.066mg/kg. The buffalo that received the 0.03mg/kg dose presented severe clinical signs but recovered six days later. The bovine that received the same dose, died after a clinical course of 22h56min. The 0.066mg/kg dose caused death of the bovine as also the buffalo, with a clinical course of 4h23min and 8h12min, respectively. The buffalo that received the 0.015mg/kg dose recovered, after a course of 48 hours. The buffalo that died, showed clinical signs from 3h58min on, and the buffaloes that showed symptoms from 17h25min and 24h00min after inoculation of the venom, but recovered. In the two cattle that died (with doses of 0.03 and 0.066mg/kg), the first clinical signs occurred 6h10min and 6h31min after the inoculation of the venom. The inoculation produced in the buffaloes and cattle nervous symptoms of flaccid paralysis. The main signs observed in the buffaloes as well as the cattle, were slight volume increase at the site of inoculation, respiration difficulties characterized by predominantly abdominal breathing, apathy, sialorreia, difficulty to get up when stimulated, evolution to sternal decubitus followed by lateral decubitus with peddling movements of the legs, and decrease of the reflexes related to the cranial nerves. The buffaloes showed also augmentation of the sustentation base, dragging of the hooves of the hind legs, slow and staggering gait, difficulty in apprehension of the food, The cattle showed additionally paralysis of the eyeballs, revealed through non-exhibition of the sclera during head rotation in latero-caudal direction. Laboratory exams revealed in the cattle and the buffaloes leucocytosis by neutrofilia, and in the biochemistry series, increase in the levels of alanine aminotransferase, aspartato aminotransferase, creatinaquinase and lactic dehydrogenase. There was no alteration in the urinalysis nor in the activation time of protrombine and in the time of partially activated tromboplastin. Necropsy only evidenced slight edema corresponding to the inoculation site in one bovine. Histopathological examination revealed picnosis of the epitelial cell nuclei of some kidney tubules in the cortex (in the buffalo and in one bovine) and slight vacuolation of hepatocites (in one bovine).

• Crotalic poisoning Crotalus durissus terrificus snakebite accident nvenenamento crotálico acidente ofídico

Abstract in Portuguese:

RESUMO.- Barbosa J.D., Sousa M.G.S., Tokarnia C.H., Brito M.F., Belo Reis A.S., Bomjardim H.A., Lopes C.T.A. & Oliveira C.M.C. 2011. [Comparison between the clinical and pathological picture in the experimental poisoning by crotalic venom.] Quadro clínico-patológico do envenenamento crotálico experimental em bubalinos comparado com o de bovinos. Pesquisa Veterinária Brasileira 31(11):967-973. Central de Diagnóstico Veterinário, Universidade Federal do Pará, Rua Maximino Porpino da Silva 1000, Pirapora, Castanhal, PA 68743-080, Brazil. E-mail: diomedes@ufpa.br O estudo teve por objetivo verificar a sensibilidade dos bubalinos à peçonha de Crotalus durissus terrificus, estudar o quadro clínico-patológico e laboratorial nessa espécie e estabelecer comparações com o verificado em bovinos. A inoculação do veneno liofilizado de Crotalus durissus terrificus, diluído em 1ml de solução fisiológica, foi feita na região da articulação úmero-rádio-ulnar, por via subcutânea, em três bubalinos (doses de 0,015; 0,03; e 0,066mg/kg) e em dois bovinos (doses de 0,03 e 0,066mg/kg). O bubalino que recebeu a dose de 0,03mg/kg apresentou sinais clínicos graves recuperou-se seis dias após, e o bovino que recebeu a mesma dose morreu com evolução de 22h56min. A dose de 0,066mg/kg causou a morte tanto do bovino quanto do bubalino, com evolução clínica de 4h23min e 8h12min, respectivamente. O bubalino que recebeu a dose de 0,015mg/kg, recuperou-se com evolução de 48 horas. Os sinais clínicos tiveram início dentro de 3h58min no bubalino que morreu, e nos bubalinos que adoeceram mas se recuperaram, dentro de 17h25min e 24h00min após a inoculação do veneno. Nos dois bovinos que morreram (com doses de 0,03 e 0,066mg/kg), os primeiros sinais clínicos foram observados 6h10min e 6h31min após a inoculação do veneno. A inoculação do veneno produziu nos búfalos e bovinos um quadro nervoso de paralisia flácida. Os principais sinais observados tanto nos búfalos quanto nos bovinos, foram discreto aumento de volume no local da inoculação, dificuldade respiratória caracterizada por respiração predominantemente abdominal, apatia, sialorreia, dificuldade para se levantar quando estimulados, evolução para decúbito esternal permanente, seguido de decúbito lateral e movimentos de pedalagem, e diminuição dos reflexos relacionados aos pares de nervos cranianos. Nos bubalinos adicionalmente foi observado aumento da base de sustentação, arrastar das pinças dos membros posteriores, marcha lenta e cambaleante, dificuldade na apreensão dos alimentos; nos bovinos ainda foram observados paralisia do globo ocular, revelada através da não exposição da esclera durante a rotação da cabeça na direção latero-caudal. Tanto nos bovinos quanto nos bubalinos, verificou-se no leucograma, leucocitose por neutrofilia, e na bioquímica sérica, aumento nos níveis de alanina aminotransferase, aspartato aminotransferase, creatinaquinase e dehidrogenase láctica. Não houve alterações na urinálise, nem no tempo de ativação da protrombina e nem no tempo de tromboplastina parcial ativada. À necropsia evidenciou-se apenas discreto edema correspondente ao local da inoculação em um bovino. Os achados histopatológicos observados foram picnose nos núcleos de células epiteliais de alguns túbulos uriníferos no córtex renal (em um búfalo e um bovino) e fígado com leve vacuolização de hepatócitos (em um bovino).

Abstract in English:

ABSTRACT.- Rodrigues R.O., Herrmann G.P., Heinemann M.B., Lage A.P., Lopes L.B. & Moreira E.C. 2011. [Comparison between immunity induced in cattle vaccinated with commercial polyvalent bacterins and one experimental monovalent.] Comparação entre a imunidade induzida em bovinos vacinados com bacterinas polivalentes comerciais e uma monovalente experimental. Pesquisa Veterinária Brasileira 31(1):10-16. Departamento de Medicina Veterinária Preventiva e Epidemiologia, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG 30123-970, Brazil. E-mail: rogerrodriguesvet@gmail.com The study evaluated the induction of antibody production against ten bacterins, nine polyvalent and one experimental monovalent to serovar Hardjo strain Norma. An indirect enzyme linked immunosorbent assay (ELISA) was developed using anti-IgG conjugate to measure total levels of IgG class antibodies conferred by bacterins using three different strains: Hardjoprajiitino, Norma and Hardjo-bovis. Microscopic Agglutination Test (MAT) was also used to measure immunoglobulin levels of the same strains. Variable ELISA titers were induced by the tested bacterins. The MAT titers found showed lower intensity and shorter duration, indicating the need to cellular control in further standardization of these vaccines. Based on results of this study, the monovalent bacterin showed best performance.

• Leptospira vaccine immunoglobulin vacina imunoglobulinas

Abstract in Portuguese:

RESUMO.- Rodrigues R.O., Herrmann G.P., Heinemann M.B., Lage A.P., Lopes L.B. & Moreira E.C. 2011. [Comparison between immunity induced in cattle vaccinated with commercial polyvalent bacterins and one experimental monovalent.] Comparação entre a imunidade induzida em bovinos vacinados com bacterinas polivalentes comerciais e uma monovalente experimental. Pesquisa Veterinária Brasileira 31(1):10-16. Departamento de Medicina Veterinária Preventiva e Epidemiologia, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG 30123-970, Brazil. E-mail: rogerrodriguesvet@gmail.com O presente estudo avaliou a indução da produção de anticorpos contra Leptospira spp.por dez bacterinas, sendo nove polivalentes e uma monovalente experimental para a sorovariedade Hardjo amostra Norma. A concentração celular foi controlada e utilizou-se adjuvante de emulsão óleo em água. Um ensaio imunoenzimático (ELISA) indireto foi desenvolvido utilizando-se conjugado anti-IgG total para mensurar os níveis de anticorpos da classe IgG conferido pelas bacterinas utilizando três amostras diferentes: Hardjoprajitino, Norma e Hardjo-bovis. Paralelamente foi utilizado também o Teste de Soroaglutinação Microscópica (SAM) para mensurar os níveis de anticorpos contra as mesmas amostras. Encontraram-se títulos variáveis entre as bacterinas de acordo com o teste ELISA. Os títulos no SAM foram de pouca intensidade e de curta duração indicando a necessidade de controle celular para uma posterior padronização destes produtos. Com base nos resultados encontrados no presente estudo, a bacterina monovalente foi a que apresentou melhor desempenho.

Abstract in English:

ABSTRACT.- Safatle A.M.V., Hvenegaard A.P., Otsuki D., Martins T.L., Kahvegian M., Berezovsky A., Salomão S.R. & Barros P.S.M. 2010. Comparison of full-field electroretinogram in diabetic and non diabetic dogs with cataracts. Pesquisa Veterinária Brasileira 30(12):1071-1076. Laboratory of Investigation in Comparative Ophthalmology, Veterinary Teaching Hospital, School of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando de Marques de Paiva 87, Bloco 8 superior, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: angsaf@usp.br Being the commonest ocular disorder, dense cataracts disable fundoscopic examination and the diagnosis of retinal disorders, which dogs may be predisposed. The aim of this study was to compare the electroretinographic responses recorded according to the International Society for Clinical Electrophysiology of Vision human protocol to evaluate retinal function of diabetic and non diabetic dogs, both presenting mature or hypermature cataracts. Full-field electroretinogram was recorded from 66 dogs, with ages varying from 6 to 15 years old allocated into two groups: (1) CG, non diabetic cataractous dogs, and (2) DG, diabetic cataractous dogs. Mean peak-to-peak amplitude (microvolts) and b-wave implicit time (milliseconds) were determined for each of the five standard full-field ERG responses (rod response, maximal response, oscillatory potentials, single-flash cone response and 30 Hz flicker). Comparing CG to DG, ERGs recorded from diabetic dogs presented lower amplitude and prolonged b-wave implicit time in all ERG responses. Prolonged b-wave implicit time was statistically significant (p< 0.05) at 30 Hz flicker (24.0 ms versus 22.4 ms). These data suggests full-field ERG is capable to record sensible alterations, such as flicker’s implicit time, being useful to investigate retinal dysfunction in diabetic dogs.

• Cataracts diabetes mellitus diabetic retinopathy retina catarata retinopatia diabética

Abstract in Portuguese:

RESUMO.- Safatle A.M.V., Hvenegaard A.P., Otsuki D., Martins T.L., Kahvegian M., Berezovsky A., Salomão S.R. & Barros P.S.M. 2010. Comparison of full-field electroretinogram in diabetic and non diabetic dogs with cataracts. [Estudo comparativo do eletrorretinograma de campo total em cães diabéticos e não diabéticos com catarata.] Pesquisa Veterinária Brasileira 30(12):1071-1076. Laboratory of Investigation in Comparative Ophthalmology, Veterinary Teaching Hospital, School of Veterinary Medicine, University of São Paulo, Av. Prof. Dr. Orlando de Marques de Paiva 87, Bloco 8 superior, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: angsaf@usp.br Catarata madura e hipermadura, alteração frequentemente observada em cães, impossibilita a visibilização do fundo do olho e provável diagnóstico de degenerações retinianas. Objetivou-se comparar as respostas retiniana de cães diabéticos e não diabéticos, ambos portadores de catarata madura ou hipermadura, com auxílio do eletrorretinograma de campo total, utilizando o protocolo da International Society for Clinical Electrophysiology of Vision. Sessenta e seis cães, com idades variando entre 6 a 15 anos de idade foram divididos em dois grupos: (1) CG, cães não diabéticos com catarata madura ou hipermadura e (2) DG, cães diabéticos com catarata madura ou hipermadura. Mensurou-se amplitude pico a pico (microvolts) e tempo de culminação da onda-b (milisegundos) para as cinco respostas do ERG (resposta de bastonetes, máxima resposta, potencial oscilatório, resposta de cones e flicker a 30Hz). Avaliando-se as respostas obtidas com o exame, o grupo de cães diabéticos apresentou menor amplitude e maior tempo de culminação da onda-b em todas as respostas. O aumento do tempo de culminação da onda-b em DG foi estatisticamente significante (p<0.05) no flicker a 30 Hz (24.0ms versus 22.4ms). ERG de campo total é capaz de registrar alterações em respostas sensíveis como o tempo de culminação da onda-b do flicker, podendo ser útil para investigar retinopatias em cães diabéticos.

Abstract in English:

ABSTRACT.- Ramos C.A.N., Araújo F.R., Souza I.I.F., Guedes Jr D.S., Oliveira R.H.M., Farias T.A., Oliveira J.B., Alves L.C. & Faustino M.A.G. 2010. [Comparison between several antigens for diagnosis of Anaplasma marginale by ELISA.] Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA. Pesquisa Veterinária Brasileira 30(1):37-41. Laboratório de Doenças Parasitárias, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE 52171-900, Brazil. E-mail: carlosanramos@yahoo.com.br Bovine anaplasmosis is a major disease in tropical and subtropical regions of the world by determine economical loss due mortality and productive reduction. The disease is caused by Anaplasma marginale, an intraerythrocytic rickettsia whose control requires, besides an efficient vaccine, the accurate identification of chronically infected cattle. Although the existence of diverse methods of diagnosis of this rickettsia, the serological methods, in particular the enzyme immunosorbent assays (ELISAs), are the most used due to its versatility and practice. However, due to the high number of antigens currently available, an evaluation becomes necessary to define which antigens present the better performance in the diagnosis of anaplasmosis. Sera from cattle positive or negative to A. marginale by PCR, and sera from cattle proceeding from Brazil and Costa Rica, were tested by ELISAs based in recombinant MSP1a, MSP2, and MSP5, a pool of the three recombinant proteins, and initial body lisate antigen (CI). Using sera from A. marginale positive cattle by PCR, the highest sensitivity was shown by CI ELISA. Nevertheless, the highest specificity, with sera from negative cattle by PCR, was shown by recombinants ELISAs. The percentiles of positive cattle from Brazil and Costa Rica were higher with CI ELISA. Reasons for such differences were discussed.

• Anaplasma marginale

Abstract in Portuguese:

RESUMO.- Ramos C.A.N., Araújo F.R., Souza I.I.F., Guedes Jr D.S., Oliveira R.H.M., Farias T.A., Oliveira J.B., Alves L.C. & Faustino M.A.G. 2010. [Comparison between several antigens for diagnosis of Anaplasma marginale by ELISA.] Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA. Pesquisa Veterinária Brasileira 30(1):37-41. Laboratório de Doenças Parasitárias, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE 52171-900, Brazil. E-mail: carlosanramos@yahoo.com.br Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática–ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI). Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.

Abstract in English:

ABSTRACT.- Ferreira Junior R.S., Siqueira A.K., Campagner M.V., Salerno T., Soares T.C.S., Lucheis S.B., Paes A.C. & Barraviera B. 2009. Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus) microbiota. [Comparação da microbiota de cascavéis (Crotalus durissus terrificus) de vida-livre e cativeiro.]Pesquisa Veterinária Brasileira 29(12):999-1003. Centro de Estudos de Venenos e Animais Peçonhentos, Universidade Estadual Paulista, Fazenda Experimental Lageado, Rua José Barbosa de Barros 1780, Botucatu, SP 18610-307, Brazil. E-mail: rseabra@cevap.org.br The study evaluated and compared the aerobic microbiota from the oral cavity, cloaca and venom of Crotalus durissus terrificus snakes, recently caught from the wild and kept under quarantine (WQ), individual captivity (IC) and collective captivity (CC). Antimicrobial drug effectiveness on isolated agents also was assayed. From group I, II and III were isolated, respectively, 29 (63.04%), 38 (90.48%) and 21 (42.86%) microorganisms from the cloaca; 15 (32.61%), 3 (7.14%) and 25 (51.02%) microorganisms from the oral cavity; and, 2 (4.35%), 1 (2.38%) and 3 (6.12%) microorganisms from venom. The most frequent bacteria were Pseudomonas aeruginosa, Proteus vulgaris and Morganella morganii, with sensitivity to amikacin, gentamicin, norfloxacin, sulfazotrin and tobramycin. Snakes kept in semi-open captivity exhibited the fewest microorganisms in oral cavities, perhaps due to the environment in captivity, with different temperature gradients, running water, absence of daily handling, circulating air, possibility of moving around, daily cleaning, and sunlight access.

• Cascavel Crotalus durissus terrificus

Abstract in Portuguese:

RESUMO.- Ferreira Junior R.S., Siqueira A.K., Campagner M.V., Salerno T., Soares T.C.S., Lucheis S.B., Paes A.C. & Barraviera B. 2009. Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus) microbiota. Pesquisa Veterinária Brasileira 29(12):999-1003. Centro de Estudos de Venenos e Animais Peçonhentos, Universidade Estadual Paulista, Fazenda Experimental Lageado, Rua José Barbosa de Barros 1780, Botucatu, SP 18610-307, Brazil. E-mail: rseabra@cevap.org.br Este estudo avaliou e comparou a microflora aeróbica da cavidade oral, cloaca e veneno de serpentes Crotalus durissus terrificus recém-capturadas da natureza e mantidas sob quarentena (WQ), mantidas em cativeiro coletivo (CC) e em cativeiro individual (IC). A eficácia de drogas antimicrobianas de agentes isolados foi também avaliada. Foram isolados microorganismos dos grupos I, II e III respectivamente: 29 (63.04%), 38 (90.48%) e 21 (42.86%) da cloaca; 15 (32.61%), 3 (7.14%) e 25 (51.02%) da cavidade oral, e finalmente 2 (4.35%), 1 (2.38%) e 3 (6.12%) do veneno. As bactérias mais frequentes foram Pseudomonas aeruginosa, Proteus vulgaris e Morganella morganii, com sensibilidade para amikacina, gentamicina, norfloxacina, sulfazotrina e tobramicina. Serpentes mantidas no cativeiro semi-aberto mostraram menor número de agentes infecciosos em cavidade oral, talvez devido ao ambiente de cativeiro com diferentes gradientes de temperatura, água corrente, ausência de manejo diário, ampla circulação de ar, possibilidade de movimentação pelos animais, limpeza diária e acesso ao Sol.

Abstract in English:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

• Stress Leopardus pardalis ocelot ACTH challenge fecal glucocorticoid jaguatirica ACTH glicocorticóides fecais

Abstract in Portuguese:

ABSTRACT.- Dias E.A., Nichi M. & Guimarães M.A.B.V. 2008. Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge. Pesquisa Veterinária Brasileira 28(7):329-334. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: eantdias@usp.br The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at -20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

Abstract in English:

ABSTRACT.- Domingues-F. L.M., Gioso M.A. & Padilha Filho J.G. 2007. Histopathological and histomicrobiological study of root canal therapy medication, comparison of calcium hydroxide versus gutta-percha with zinc oxide/eugenol in the teeth of dogs. Pesquisa Veterinária Brasileira 27(2):71-74. Departamento de Cirurgia da Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando de Marques de Paiva 87, Bloco 8-superior, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: lesliedf@usp.br The presence of microorganisms in dental structures with experimentally induced necrosis was evaluated. The materials were tested to evaluate their antimicrobial activity and tissue repair efficacy. Four dogs were used in this experiment, with a total of 64 roots of premolar teeth, divided into three groups. The root canals of Group I were filled with gutta-percha and zinc oxide/eugenol cement; Group II were filled with calcium hydroxide, and Group III were not filled. All animals were clinically and radiographically examined 15 days after surgery and then again every subsequent 15 days until 120 days, when the teeth were extracted en bloc. Histopathological analysis showed inflammatory infiltration, cement and bone resorption and necrotic tissue in the apical delta in different proportions. Histomicrobiological analysis showed the presence of microorganisms inside the teeth structures, with different concentrations according to the treatment used. There was statistical significance between the groups (p>0.05). Gutta-percha with zinc oxide/eugenol demonstrated good antimicrobial activity; calcium hydroxide was not efficient. The conclusion of this study is that gutta-percha with zinc oxide/eugenol is the better protocol for filling root canals in dogs.

• Apical delta endodontic gutta-percha calcium hidroxide microorganisms dogs

Abstract in Portuguese:

ABSTRACT.- Domingues-F. L.M., Gioso M.A. & Padilha Filho J.G. 2007. Histopathological and histomicrobiological study of root canal therapy medication, comparison of calcium hydroxide versus gutta-percha with zinc oxide/eugenol in the teeth of dogs. Pesquisa Veterinária Brasileira 27(2):71-74. Departamento de Cirurgia da Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando de Marques de Paiva 87, Bloco 8-superior, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: lesliedf@usp.br The presence of microorganisms in dental structures with experimentally induced necrosis was evaluated. The materials were tested to evaluate their antimicrobial activity and tissue repair efficacy. Four dogs were used in this experiment, with a total of 64 roots of premolar teeth, divided into three groups. The root canals of Group I were filled with gutta-percha and zinc oxide/eugenol cement; Group II were filled with calcium hydroxide, and Group III were not filled. All animals were clinically and radiographically examined 15 days after surgery and then again every subsequent 15 days until 120 days, when the teeth were extracted en bloc. Histopathological analysis showed inflammatory infiltration, cement and bone resorption and necrotic tissue in the apical delta in different proportions. Histomicrobiological analysis showed the presence of microorganisms inside the teeth structures, with different concentrations according to the treatment used. There was statistical significance between the groups (p>0.05). Gutta-percha with zinc oxide/eugenol demonstrated good antimicrobial activity; calcium hydroxide was not efficient. The conclusion of this study is that gutta-percha with zinc oxide/eugenol is the better protocol for filling root canals in dogs.

Abstract in English:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

• Leptospires diagnostic techniques Mus musculus lesions.

Abstract in Portuguese:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

Abstract in English:

ABSTRACT.- Barbosa]. D., Oliveira C. M. C., Tokarnia C. H. & Riet-Correa E 2003. [Comparison of the sensibility of cattle and buffaloes to poisoning by Palicourea marcgravii (Rubiaceae).] Comparação da sensibilidade de bovinos e búfalos à intoxicação por Palicourea marcgravii (Rubiaceae). Pesquisa Veterinária Brasileira 23(4): 167-172. Escola de Medicina Veterinária, Campus Castanhal, Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68743- 080, Brazil. E-mail: diomedes@ufpa.br In order to verify if buffaloes are more resistant than cattle to poisoning by Palicourea marcgravii, the plant was given per os simultaneously to buffaloes and cattle. Seven buffaloes and three head of cattle were used. Doses of 0.5, 1.0 and 2.0 g/kg did not cause signs of poisoning in buffaloes; but doses of 3.0, 4.0 and 6.0 g/kg caused death. In cattle 0.25 g/kg did not cause symptoms; but 0.5 and 2.0 g/kg were lethal. The influence of exercise on the onset of symptoms, the time which elapsed from the beginning of the administration of the plant to the appearance of symptoms, and the symptoms themselves, were similar in the two animal species. The clinical course was longer in buffaloes; whilst in cattle the period between the onset of severe symptoms and death lasted 9 to 17 minutes; in buffaloes it varied from 10 minutes to 1 hour 28 minutes. It can be concluded, that buffaloes are about six times more resistant than cattle to the toxic action of P. maracgravii. The lower incidence of deaths in buffaloes by poisonous plants in the Amazon region is, at least in part, due the lower sensibility of buffaloes to poisoning by this plant. Another factor responsible for the smaller number of buffaloes which die due to plant poisoning in the Amazon region, is possibly that buffaloes prefer the Jowlands, e.g. temporarily flooded areas, which are the habitat of Arrabidaea bilabiata, the second most important poisonous plant of the Amazon region. This plant is less poisonous than P. marcgravii, and its habitat are the higher not flooded areas. So it would be safer, in order to diminish losses, to breed buffaloes instead of cattle in areas where P. marcgravii occurs. The reason for the greater resistance of buffaloes should be further investigated as a possible prophylactic measure to avoid poisoning by P. marcgravii in cattle. On the other hand, Australian investigators modified genetically the ruminai bacteria Butyrivibrio fibrisolvens by introducing a gene, isolated from a Moraxella sp, which codifies a dehalogenase, capable of hydrolyzing fluoroacetate, the poisonous principle of P. marcgravii. Transferring the genetically modified B. fibrisolvens into the rumen of animal, which consume plants that contain fluoroacetate, would be a viable method to control that poisoning through a ruminal detoxifying process for the toxin. Through preliminary contacts, the director of the consorcium in Australia, responsible for the modification of the bacteria, declared that he is interested to sell the technology to our country, but it would be necessary to know if it is permitted to introduce into Brazil the genetically modified bacteria. If allowed, it would be necessary to perform investigations on the viability and the methodology for using this bacteria to control P. marcgravii poisoning in our country.

• Poisonous plants plant poisoning Palicourea marcgravii Rubiaceae cattle buffaloes prophylaxis Butyrivibrio fibrisolvens genetically modified bactéria Intoxicação por planta Rubiaceae bovinos búfalos profilaxia bactérias geneticamente modificadas.

Abstract in Portuguese:

RESUMO.- Barbosa]. D., Oliveira C. M. C., Tokarnia C. H. & Riet-Correa E 2003. [Comparison of the sensibility of cattle and buffaloes to poisoning by Palicourea marcgravii (Rubiaceae).] Comparação da sensibilidade de bovinos e búfalos à intoxicação por Palicourea marcgravii (Rubiaceae). Pesquisa Veterinária Brasileira 23(4): 167-172. Escola de Medicina Veterinária, Campus Castanhal, Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68743- 080, Brazil. E-mail: diomedes@ufpa.br O presente estudo foi realizado para verificar se búfalos são mais resistentes do que bovinos à ação tóxica de Palicourea marcgravii, mediante a administração da planta por via oral, simultaneamente, a bovinos e a búfalos. Foram usados sete búfalos e três bovinos. Verificou-se que em búfalos doses de 0,5 g/kg, 1,0 g/kg e 2,0 g/kg não causaram sintomas de intoxicação. As doses de 3,0 g/kg, 4,0 g/kg e 6,0 g/kg causaram a morte dos búfalos. Em bovinos, a dose de 0,25g/kg não causou sinais clínicos de intoxicação, enquanto que doses de 0,5 g/kg e 2,0 g/kg causaram a morte. A influência do exercício sobre o aparecimento dos sintomas, o prazo decorrido desde o começo da administração da planta até o início de sintomas, e os próprios sintomas, foram semelhantes nas duas espécies animais. O curso clínico foi mais longo nos búfalos. Enquanto nos bovinos o período entre o aparecimento dos sintomas graves e a morte foi de 9 a 17 minutos, nos búfalos variou de 10 minutos a 1 hora e 28 minutos. Pode se concluir que os bubalinos são aproximadamente seis vezes mais resistentes do que os bovinos à ação tóxica de P. marcgravii. O menor índice de mortes pela ação de plantas tóxicas na Amazônia em búfalos é, pelo menos em parte, devido à maior resistência do búfalo à intoxicação por essa planta. Outro fator responsável pelo menor número de mortes em búfalos pela intoxicação por plantas, na Amazônia, poderia ser que os búfalos preferem a várzea, que é o habitat de Arrabidaea bilabiata, a segunda planta tóxica mais importante da Amazônia, menos tóxica do que P. marcgravii, e com habitat na terra firme. Em áreas onde ocorre P. marcgravii seria mais prudente, para diminuir os prejuízos, criar búfalos em lugar de bovinos. A causa dessa maior resistência do búfalo merece ser investigada para a eventual elaboração de métodos profiláticos da intoxicação por P. marcgravii em bovinos. Por outro lado, pesquisadores australianos modificaram geneticamente a bactéria ruminal Butyrivibrio fibrisolvens, mediante a introdução de um gene, isolado de Moraxella sp, que codifica uma dehalogenase, capaz de hidro-lizar fluoroacetato. A transferência de B.fibrisolvens geneticamente modificado para o rúmen de animais ingerindo plantas que contêm fluoroacetato, como é o caso de P. marcgravii, seria um método viável para o controle da intoxicação mediante a detoxificação ruminal do princípio ativo. Em contatos preliminares o diretor responsável do consórcio na Austrália responsável pela modificação da bactéria, declarou o interesse em vender a tecnologia ao nosso país, porém seria necessário saber se é possível importar essa bactéria geneticamente modificada no Brasil. Caso positivo, seria indispensável realizar pesquisas sobre a viabilidade e a metodologia para o uso dessa bactéria em nosso meio.