PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

The histological and molecular analysis of biopsy samples are fundamental steps for the understanding of physiopathology, diagnosis and prognosis of the diseases. However, harvest of tissue biopsies from hoof lamellar tissue is a procedure with limitations due to lack of effective surgical instruments and techniques. The aim of the current study is to develop and test in vivo a surgical instrument with the specific purpose of harvesting lamellar tissue in cattle. A prototype called Falcão-Faleiros’ lamellotome (INPIBR102013018765-8) was designed, produced and tested. After sedation, five adult cattle were restrained in lateral recumbency and locally anesthetized in two digits. The stratum corneum was worn down using a rotary tool coupled to a 3/8” high-speed cutter until the soft tissue proximity was reached. Next, the inner edge of the worn area was bounded with a scalpel. The lamellotome was introduced to obtain and hold the sample. The histological specimens of 16mm length by 6mm depth were stained with HE, PAS, Masson’s thricome and Shorr. The structures of interest were differentiated in the histological analysis without technical artifacts and a mean number of 85 epidermal laminae per sample were counted. No relevant lameness or wound complication were seen following the procedure. In conclusion the technique using the lamellotme was effective in obtaining lamellar tissue biopsy samples without causing clinical harm in cattle. The procedure showed potential to be used in clinical research or even as a supplementary diagnosis method for routine bovine podiatry.

• Surgery instrument bovine hoof biopsy claw laminitis cattle

Abstract in Portuguese:

A avaliação das propriedades histológicas e da expressão de genes e proteínas em biópsias tem sido determinante para o entendimento da fisiopatologia, o diagnóstico e o prognóstico das enfermidades. Entretanto, a obtenção de biópsias do casco é um procedimento com limitações devido à ausência de técnicas e instrumentos específicos. O objetivo foi desenvolver e testar, na espécie bovina, um instrumento cirúrgico especificamente desenvolvido para realização de biópsias de casco nominado lamelótomo de Falcão-Faleiros (INPI, BR102013018765-8). Utilizaram‑se cinco bovinos adultos que foram sedados, contidos em decúbito lateral e tiveram dois dígitos anestesiados. Em seguida, uma serra circular acoplada a uma microretífica foi usada para o desgaste do estrato córneo na parede dorsal até próximo do estrato lamelar. Após incisões retilíneas delimitando a borda interna da área desgastada, utilizou-se o lamelótomo para obtenção da amostra. Os fragmentos de 16mm de comprimento e 6mm de profundidade foram fixados em formalina e processados para histologia com colorações HE, PAS, Shorr e tricrômico de Masson. Nenhum dos animais apresentou claudicação ou complicação relevantes no período pós-opertório. As amostras foram consideradas adequadas quanto à integridade das lâminas e à preservação de sua arquitetura. Obtiveram-se média de 85 lâminas epidérmicas viáveis por biópsia. Conclui-se que o lamelótomo de Falcão-Faleiros é apropriado e seguro para a obtenção de biópsias de casco em bovinos, se mostrando promissor para uso em estudos clínicos e na rotina de diagnóstico de problemas podais em bovinos.

• Instrumento cirúrgico biópsia tecido laminar bovinos casco laminite cirurgia

Abstract in English:

ABSTRACT.- Carvalho T.F., Pereira N.B., Rocha C.R.J., Figueiredo C.C., Rachid M.A. & Vasconcelos A.C. 2016. [Apoptosis and maturation of the bovine placenta: an immunohistochemical and morphometric study.] Apoptose e maturação placentária bovina: um estudo imuno-histoquímico e morfométrico. Pesquisa Veterinária Brasileira 36(3):237-246. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: carvalhotativet@hotmail.com Placental release after birth involves loss of maternal-fetal adhesion and occurs only after complete maturation of the placentoma related to the decrease in cellularity of fetal and maternal tissues. Apoptosis is required for both the normal maturation and release of the placenta after birth. The aim of this study was to evaluate the occurrence of apoptosis in samples of the placenta of cows in different stages of gestation. Samples of 15 healthy cow placentomas at 4 (n=5), 6 (n=5) and 9 (n=5) months of gestation were harvested and processed for routine histology, immunohistochemistry and histochemistry. The slides were stained with HE, PicroSirius Red and subjected to immunohistochemical analysis of proteins Caspase 3, Caspase 8, Bax and Bid. Increase in number of vessels was not associated with increase in vascular area during progression of gestation. The results of histomorphometry revealed increased labeling for Bax and Caspases 3 and 8 in trophoblastic binucleated cells in late pregnancy, where the Bid remained without significant change. Histomorphometry analyzing the mononuclear trophoblast cells showed a high expression for Bax in early pregnancy, but decreased at 6 months of gestation. Immunolabeling revealed increased Caspases 3/8 and Bid with advancing of gestation. Further evaluation of type I and III collagen showed a decrease of both types of collagens at the end of gestation, what is very important for the reduction of maternal-fetal adhesion. These results confirm that Caspases 3 and 8 and Bax are involved in the mechanisms of activation of apoptosis through mitochondrial intrinsic and/or extrinsic pathway during pregnancy in trophoblastic binucleated cells. In mononuclear trophoblast cells Bax looses importance in the apoptosis process, awhile Bid and caspases 3 and 8 become the most significant.

• Placenta

Abstract in Portuguese:

RESUMO.- Carvalho T.F., Pereira N.B., Rocha C.R.J., Figueiredo C.C., Rachid M.A. & Vasconcelos A.C. 2016. [Apoptosis and maturation of the bovine placenta: an immunohistochemical and morphometric study.] Apoptose e maturação placentária bovina: um estudo imuno-histoquímico e morfométrico. Pesquisa Veterinária Brasileira 36(3):237-246. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: carvalhotativet@hotmail.com A liberação da placenta após o parto envolve a perda da adesão materno-fetal e ocorre somente após a maturação completa do placentoma, que está relacionada com a diminuição da celularidade dos tecidos fetal e materno. A apoptose é requerida tanto para a maturação quanto para a liberação normal da placenta após o parto. O objetivo do presente estudo foi avaliar a ocorrência de apoptose em amostras de placenta de vacas em diferentes fases de gestação. Amostras de placentomas de 15 vacas saudáveis com 4 (n=5), 6 (n=5) e 9 (n=5) meses de gestação foram colhidas e processadas rotineiramente para a histologia, imunoistoquímica e histoquímica. As lâminas obtidas foram coradas em HE, Picrosirius Red e submetidas à análise imunoistoquímica das proteínas Caspase 3, Caspase 8, Bax e Bid. O aumento no número de vasos não necessariamente se associou ao aumento do calibre destes durante a evolução da gestação. Os resultados de histomorfometria revelaram aumento da marcação para Bax e Caspases 3 e 8 em células trofoblásticas binucleadas no final da gestação, enquanto o Bid se manteve sem alteração significativa. A histomorfometria das células trofoblásticas mononucleadas revelou expressão alta para Bax no início de gestação, com diminuição aos 6 meses de gestação e aumento das imunomarcações para Caspases 3 e 8, e Bid com o avanço gestacional. Os colágenos tipo I e III não aumentaram do terço médio ao final da gestação, o que é importante para a diminuição da adesão materno-fetal. Esses resultados confirmam que as Caspases 3 e 8, e o Bax estão envolvidos nos mecanismos de ativação da apoptose pela via intrínseca mitocondrial e/ou extrínseca ao longo da gestação em células trofoblásticas binucleadas, e que nas células trofoblásticas mononucleadas o Bax deixa de ser importante, enquanto o Bid e as Caspases 3 e 8 se tornam os mais significativos.

Abstract in English:

ABSTRACT.- Mendes H.M.F., Casagrande F.P., Lima I.R., Souza C.H., Gontijo L.D’., Alves G.E.S., Vasconcelos A.C. & Faleiros R.R. 2013. Histopathology of dairy cows’ hooves with signs of naturally acquired laminitis. Pesquisa Veterinária Brasileira 33(5):613-619. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Campus de Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: faleirosufmg@gmail.com The purpose of this study was to investigate histological changes in dairy cows’ hooves with or without injuries from naturally acquired laminitis. Cull cows with no clinical signs of hoof abnormalities (G1, n=9) and those with macroscopic lesions associated with laminitis without (G2, n=23) or with lameness (G3, n=7) were used in the study. After slaughter, samples of dermo-epidermal junctions of sole, axial and dorsal regions of the hoof were obtained and histologically processed using HE and PAS staining. Congestion, hemorrhage and inflammatory infiltrate in the dermis of sole, axial and dorsal regions were blindly and semiquantitatively evaluated by the same researcher. Inflammatory infiltrate was evaluated in the dermal laminae of axial and dorsal regions. The morphology of epidermal cells and the presence of irregularities in three regions of the basement membrane (BM) length were examined using PAS staining. Scores of lesions in different regions of the hoof in the same group and in different groups for each region of the hoof were compared using non-parametric analyses. Inflammatory infiltrate in the dermis of all regions of the hoof was detected in all groups with no significant statistical difference. Cows with no clinical signs of hoof abnormalities secondary to laminitis (G1) have inflammation scores and epidermal cell changes similar to those of groups with laminitis injuries, suggesting the existence of a prodromal phase for this disease in bovines. BM had irregularities with a variable intensity along its length, however, with no difference among groups. The pattern of BM irregularities found has not been reported so far and does not resemble the BM collapse described in horses and cattle with induced acute laminitis. Is it concluded that even in the absence of macroscopic hoof signs associated to laminitis, dairy cows have histological injuries compatible with inflammation of the dermo-epidermal junction as in affected animals. Basement membrane of cows with or without laminitis associated lesions had irregularities with an irregular distribution along its length which need to be further studied.

• Laminitis lameness hoove

Abstract in Portuguese:

RESUMO.- Mendes H.M.F., Casagrande F.P., Lima I.R., Souza C.H., Gontijo L.D’., Alves G.E.S., Vasconcelos A.C. & Faleiros R.R. 2013. Histopathology of dairy cows’ hooves with signs of naturally acquired laminitis. [Histopatologia do casco de vacas de aptidão leiteira com e sem sinais de laminite naturalmente adquirida.] Pesquisa Veterinária Brasileira 33(5):613-619. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Campus de Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: faleirosufmg@gmail.com O objetivo deste trabalho foi investigar as alterações histológicas no casco de vacas leiteiras sem e com lesões de laminite, naturalmente, adquirida. Utilizaram-se animais de descarte sem lesões macroscópicas no casco (G1 n=9) e com lesões macroscópicas associadas à laminite sem (G2 n=23), ou com claudicação (G3 n=7). Após o abate, amostras da junção derme-epiderme das regiões solear, axial e dorsal do casco foram obtidas e processadas, histologicamente, nas colorações de HE e PAS. Avaliou-se, semiquantitativamente, às cegas e por um mesmo pesquisador, congestão, hemorragia e infiltrado inflamatório na derme das regiões solear, axial e dorsal. Avaliou-se infiltrado inflamatório na lâmina dermal das regiões axial e dorsal. A morfologia das células epidermais e a presença de irregularidades em três regiões do comprimento da membrana basal (MB) foram examinadas na coloração de PAS. Os escores das lesões nas diferentes regiões do casco no mesmo grupo e nos diferentes grupos para cada região do casco foram comparados através de análises não paramétricas (P<0,05). Infiltrado inflamatório na derme de todas as regiões do casco foi detectado em todos os grupos sem diferença estatística (P>0,05). Vacas sem lesões macroscópicas secundárias à laminite (G1) apresentaram escores de inflamação e alteração de células epidermais semelhantes aos dos grupos com lesões de laminite, sugerindo a existência de fase prodrômica para a doença em bovinos. A MB apresentou irregularidades com intensidade variável ao longo de seu comprimento (P<0,05), porém, sem diferença entre grupos (P>0,05). O padrão de irregularidades na MB encontrado não foi relatado até o momento e não se assemelha ao colapso de MB descrito em equinos e bovinos com laminite induzida. Concluiu-se que, mesmo na ausência de lesões macroscópicas no casco causadas por laminite, vacas leiteiras apresentam lesões histológicas compatíveis com inflamação da junção derme-epiderme como em animais afetados. A membrana basal de bovinos sem e com lesões de laminite apresenta irregularidades com distribuição irregular ao longo de seu comprimento que precisam ser melhor estudadas.

Abstract in English:

ABSTRACT.- Meça K.K.O.L., Del Puerto H.L., Rodrigues L.V., Rachid M.A., Pereira N.B., Cândido M.G.L. & Vasconcelos A.C. 2011. [Apoptosis in placental maturation of cows in different stages of pregnancy: Immunohistochemical and biochemical evidences.] Apoptose na maturação placentária de vacas em diferentes estágios de gestação: evidenciação imuno-histoquímica e bioquímica. Pesquisa Veterinária Brasileira 31(8):718-722. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Campus Pampulha, BeloHorizonte, MG 31270-010, Brazil. E-mail: karinameca@yahoo.com.br Apoptosis is important for placental homeostasis maintenance, and a misbalance in this process may compromise the success of pregnancy. The aim of this study is to evaluate apoptosis in bovine placental samples at different stages of pregnancy. Placentome samples from 15 healthy cows, at gestational ages of 4 (n=5), 6 (n=5) and 9 (n=5) months were collected and routinely processed for histological and immunohistochemical analysis, and for DNA isolation. Histopathology sections were stained with HE. Others were submitted to immunohistochemistry, using pro-apoptotic caspase-3 and Bax protein, and anti-apoptotic Bcl-2 specific antibodies. Isolated DNAs were processed for electrophoresis in agarose gel, for detection of internucleosomal fragmentation. Histomorphometry results demonstrated that apoptotic cells gradually increase with the advance of the gestation. Also, the DNA ladder pattern was observed in all groups. In addition, caspase-3, Bax and Bcl-2 were all expressed in the three different gestation periods. However, caspase-3 presented a higher expression in all groups, in comparison to Bcl-2 and Bax. These results confirm the importance of the apoptosis in the placental maturation. Also, these results indicated that caspase-3, Bcl-2 and Bax are involved in apoptotic activation mechanisms by mitochondrial intrinsic pathway during placental maturation, contributing for physiological cellularity and cellular turn over balance in bovine placenta.

• Apoptosis placenta apoptose

Abstract in Portuguese:

RESUMO.- Meça K.K.O.L., Del Puerto H.L., Rodrigues L.V., Rachid M.A., Pereira N.B., Cândido M.G.L. & Vasconcelos A.C. 2011. [Apoptosis in placental maturation of cows in different stages of pregnancy: Immunohistochemical and biochemical evidences.] Apoptose na maturação placentária de vacas em diferentes estágios de gestação: evidenciação imuno-histoquímica e bioquímica. Pesquisa Veterinária Brasileira 31(8):718-722. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Campus Pampulha, BeloHorizonte, MG 31270-010, Brazil. E-mail: karinameca@yahoo.com.br Apoptose tem um papel importante na manutenção da homeostase placentária, e o desequilíbrio desse processo pode comprometer a gestação. O objetivo do presente estudo foi avaliar a ocorrencia de apoptose em amostras de placenta de vacas em diferentes fases de gestação. Amostras de placentomos de 15 vacas saudáveis com 4 (n=5), 6 (n=5) e 9 (n=5) meses de gestação foram colhidas e processadas rotineiramente para a histologia, imuno-histoquímica e isolamento de DNA. As lâminas obtidas foram coradas em HE, ou submetidas à análise imuno-histoquímica das proteínas pró-apoptóticas caspase-3 e Bax, e da proteína anti-apoptótica Bcl-2. O DNA isolado foi submetido à eletroforese em gel de agarose para detecção da fragmentação internucleossômica do genoma. Os resultados de histomorfometria revelaram que as células apoptóticas aumentaram progressivamente com o avanço da gestação. Confirmou-se a apoptose pela fragmentação característica do DNA genômico, visualizada pelo clássico “padrão em escada” na eletroforese em gel de agarose. Adcionalmente, a imunoexpressão de caspase-3, Bax e Bcl-2 foram observadas em todas as amostras. Entretanto, a proteína caspase-3 apresentou marcação mais intensa em todos os tempos gestacionais, quando comparada com a marcação das proteínas Bcl-2 e Bax. Esses resultados confirmam e reforçam a importância da apoptose na maturação placentária. Além disto, indica que caspase-3, Bax e Bcl-2 estão envolvidas nos mecanismos de ativação da apoptose pela via intrínseca mitocondrial ao longo da gestação, contribuindo para o equilíbrio fisiológico da celularidade e renovação celular na placenta bovina.

Abstract in English:

ABSTRACT.- Alves G.E.S., Mendes H.M.F., Alves T.G.S., Faleiros R.R., Vasconcelos A.C., Santos R.L. & Rio Tinto J.J.M. 2011. Hydrocortisone decreases apoptosis in jejunum of horses subjected to experimental ischemia and reperfusion. Pesquisa Veterinária Brasileira 31(6):471-476. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil. E-mail: geraldo@vet.ufmg.br In order to evaluate the effect of hydrocortisone on apoptosis in the jejunum of horses subjected to ischemia and reperfusion, ten horses were paired and grouped into two groups - treated (n=5) and non treated (n=5). Segments of the jejunum were used as controls (C), or as venous ischemia (VIsc), which were subjected to 2h of ischemia followed by 2 or 12h of reperfusion. C samples were collected at time zero (prior to ischemia) and VIsc samples were collected at 2h of ischemia and at 2 and 12h of reperfusion. TUNEL positive apoptotic cells were counted in 10 microscopical fields in deep mucosa from each horse throughout the time course. After 12h of reperfusion, the number of apoptotic cells in treated group were significantly lower than in untreated animals, indicating that hydrocortisone inhibits apoptosis. These results indicate that hydrocortisone has a beneficial effects favoring the maintenance of jejunal integrity in horses with ischemia and reperfusion injuries by preventing apoptotic cell death.

• Hydrocortisone ischemia reperfusion apopitosis isquemia reperfusão apopitose

Abstract in Portuguese:

RESUMO.- Alves G.E.S., Mendes H.M.F., Alves T.G.S., Faleiros R.R., Vasconcelos A.C., Santos R.L. & Rio Tinto J.J.M. 2011. Hydrocortisone decreases apoptosis in jejunum of horses subjected to experimental ischemia and reperfusion. [Hidrocortisona diminui a apoptose no jejuno de equinos sujeitos a isquemia e reperfusão experimentais.] Pesquisa Veterinária Brasileira 31(6):471-476. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil. E-mail: geraldo@vet.ufmg.br Com o objetivo de avaliar o efeito da hidrocortisona sobre a apoptose no jejuno de equinos submetidos à is-quemia e reperfusão, dez cavalos foram agrupados em dois grupos: tratado (n=5) e não-tratado (n=5). Foi utilizado um segmento do jejuno como controle (C) e outro foi submetido a isquemia venosa (VIsc) por 2h seguida de 2 ou 12 h de re-perfusão. Amostras de C foram coletadas no tempo zero (antes da isquemia) e amostras de VIsc foram coletadas após 2h de isquemia e a 2 e 12h de reperfusão. Células apoptóticas TUNEL positivas foram contadas em 10 campos microscópicos da mucosa na região das criptas de cada animal em cada tempo. Após 12h de reperfusão, o número de células apoptóticas no grupo tratado foram significativamente menores do que no grupo não-tratado, indicando que a hidrocortiso-na inibe a apoptose. Esses resultados mostram que a hidro-cortisona tem efeito benéfico favorecendo a manutenção da integridade do jejuno em cavalos com lesão de isquemia e reperfusão por prevenir a morte celular por apoptose.

Abstract in English:

ABSTRACT.- Del Puerto H.L., Vasconcelos A.C., Moro L., Alves F., Braz G.F. & Martins A.S. 2010. Canine distemper virus detection in asymptomatic and non vaccinated dogs. Pesquisa Veterinária Brasileira 30(2):139-144. Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: helendelpuerto@hotmail.com A quantitative real time polymerase chain reaction (PCR) revealed canine distemper virus presence in peripheral blood samples from asymptomatic and non vaccinated dogs. Samples from eleven domestic dogs with no signs of canine distemper and not vaccinated at the month of collection were used. Canine distemper virus vaccine samples in VERO cells were used as positive controls. RNA was isolated with Trizol®, and treated with a TURBO DNA-free kit. Primers were designed for canine distemper virus nucleocapsid protein coding region fragment amplification (84 bp). Canine b-actin (93 bp) was utilized as the endogenous control for normalization. Quantitative results of real time PCR generated by ABI Prism 7000 SDS Software showed that 54.5% of dogs with asymptomatic canine distemper were positive for canine distemper virus. Dissociation curves confirmed the specificity of the real time PCR fragments. This technique could detect even a few copies of viral RNA and identificate subclinically infected dogs providing accurate diagnosis of this disease at an early stage.

• Cinomose cinomose canina

Abstract in Portuguese:

RESUMO.- Del Puerto H.L., Vasconcelos A.C., Moro L., Alves F., Braz G.F. & Martins A.S. 2010. Canine distemper virus detection in asymptomatic and non vaccinated dogs. Pesquisa Veterinária Brasileira 30(2):139-144. [Detecção do vírus da cinomose canina em cães assintomáticos e não vacinados.]Departamento de Patologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Campus Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: helendelpuerto@hotmail.com A reação em cadeia da polimerase (PCR) em tempo real revelou a presença do vírus da cinomose canina em amostra de sangue de cães assintomáticos e não vacinados. Amostra de onze cães domésticos sem nenhum sinal clínico de cinomose e que não foram vacinados no mês da coleta de sangue foram utilizados para análise. Amostra vacinal do vírus da cinomose canina em células VERO foi utilizada como controle positivo. O RNA total foi isolado utilizando-se Trizol®, e tratadas com o Kit TURBO DNA-free. Os iniciadores foram desenhados para amplificar a região do nucleocapsídeo viral com 319pb e 84pb para a PCR convencional e PCR em tempo real, respectivamente. O fragmento alvo da b-actina canina com 93pb foi utilizado como controle endógeno e normalizador. Resultados quantitativos da PCR em tempo real gerados pelo programa ABI Prism 7000 SDS demonstraram que 54,5% dos cães assintomáticos foram positivos para o vírus da cinomose canina. As curvas de dissociação confirmaram a especificidade dos fragmentos da PCR em tempo real. A detecção precoce do RNA viral é importante para a identificação de cães subclinicamente infectados e limitar a difusão da doença.

Abstract in English:

ABSTRACT.- Mendes H.M.F., Escobar A., Vasconcelos A.C., Zucoloto S., Alves G.E.S. & Faleiros R.R. 2009. [Morphometrical alterations in myoenteric plexus of experimentally distended equine small colon.] Alterações morfométricas no plexo mioentérico do cólon menor equino distendido experimentalmente. Pesquisa Veterinária Brasileira 29(7):557-562. Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG 30161-970, Brazil. E-mail: faleiros@ufmg.br The equine small colon is frequently affected by obstruction, and intestinal motility dysfunction is a common complication after its surgical treatment. This fact may be related to myoenteric plexus lesion caused by distention; however, little is known about the pathophysiology of this condition. The objective of this study was to evaluate the morphological alterations in the myoenteric inervation of segments of small colon of horses subjected to intraluminal distension with reduction of the microvascular perfusion (partial ischemia) of the intestinal wall. Nine horses were used to promote distension of on segment of small colon for 4 hours. Samples of intestinal wall were collected before and at the end of the distension, after 1.5 and 12 hours of reperfusion in the experimental segment and at the end of the procedure in a different distant segment. Samples were processed and histological sections were stained with cresyl violet for the morphometric studies. An image analyzer software was used to measure perimeter, diameter, and area of the neuronal body, nucleus and nucleolus of the neurons and the areas of the cytoplasm and nucleoplasm. Significant reductions (P<0.05) in the areas of the neuronal body and cytoplasm were detected at the end of intestinal distension, returning to the basal values during the reperfusion. In conclusion, intraluminal distension promoted changes in the morphology of the neurons of myoenteric plexus. These morphological modifications may be associated to the motility dysfunction frequently observed in clinical cases.

• Small colon distension myoenteric plexus horse

Abstract in Portuguese:

ABSTRACT.- Mendes H.M.F., Escobar A., Vasconcelos A.C., Zucoloto S., Alves G.E.S. & Faleiros R.R. 2009. [Morphometrical alterations in myoenteric plexus of experimentally distended equine small colon.] Alterações morfométricas no plexo mioentérico do cólon menor equino distendido experimentalmente. Pesquisa Veterinária Brasileira 29(7):557-562. Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG 30161-970, Brazil. E-mail: faleiros@ufmg.br The equine small colon is frequently affected by obstruction, and intestinal motility dysfunction is a common complication after its surgical treatment. This fact may be related to myoenteric plexus lesion caused by distention; however, little is known about the pathophysiology of this condition. The objective of this study was to evaluate the morphological alterations in the myoenteric inervation of segments of small colon of horses subjected to intraluminal distension with reduction of the microvascular perfusion (partial ischemia) of the intestinal wall. Nine horses were used to promote distension of on segment of small colon for 4 hours. Samples of intestinal wall were collected before and at the end of the distension, after 1.5 and 12 hours of reperfusion in the experimental segment and at the end of the procedure in a different distant segment. Samples were processed and histological sections were stained with cresyl violet for the morphometric studies. An image analyzer software was used to measure perimeter, diameter, and area of the neuronal body, nucleus and nucleolus of the neurons and the areas of the cytoplasm and nucleoplasm. Significant reductions (P<0.05) in the areas of the neuronal body and cytoplasm were detected at the end of intestinal distension, returning to the basal values during the reperfusion. In conclusion, intraluminal distension promoted changes in the morphology of the neurons of myoenteric plexus. These morphological modifications may be associated to the motility dysfunction frequently observed in clinical cases.

Abstract in English:

ABSTRACT.- Costa E.A., Bastianetto E., Vasconcelos A.C., Bomfim M.R.Q., Fonseca F.G., Gomes A.D., Leite R.C. & Resende M. 2009. An outbreak of malignant catarrhal fever in Murrah buffaloes in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 29(5):395-400. Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG 31270-901, Brazil. E-mail: lvc@icb.ufmg.br An outbreak of Malignant Catarrhal Fever (MCF) resulted in death of five female buffaloes and one domestic cow from the same farm. Four buffaloes died 10-15 days after the appearance of clinical signs, while the fifth was euthanized in extremis, after similar clinical signs. Histopathological lesions included multifocal histiolymphocytic epicarditis, myocarditis and lymphocytic interstitial pneumonia, which are commonly seen in cases of MCF in buffaloes. Furthermore, lymphocytic vasculitis centered in the adventitia, with occasional fibrinoid necrosis in the muscular layer, was found in the kidneys, liver, spleen, lymph nodes and brain. Nucleotide sequencing of DNA fragments from the central nervous system amplified by PCR revealed 98% similarity with known OHV-2 sequences from Genbank. Additionally, PCR analysis also revealed the presence of OHV-2 DNA in the peripheral mononuclear blood cells of two clinically healthy buffaloes. The diagnosis of MCF was based on epidemiological, clinical, gross and histopathological findings and on the results of a semi-nested PCR followed by nucleotide sequencing.

• Diseases of buffaloes doenças de búfalos gammaherpesviruses gammaherpesvirus malignant catarrhal fever

Abstract in Portuguese:

ABSTRACT.- Costa E.A., Bastianetto E., Vasconcelos A.C., Bomfim M.R.Q., Fonseca F.G., Gomes A.D., Leite R.C. & Resende M. 2009. An outbreak of malignant catarrhal fever in Murrah buffaloes in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 29(5):395-400. Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG 31270-901, Brazil. E-mail: lvc@icb.ufmg.br An outbreak of Malignant Catarrhal Fever (MCF) resulted in death of five female buffaloes and one domestic cow from the same farm. Four buffaloes died 10-15 days after the appearance of clinical signs, while the fifth was euthanized in extremis, after similar clinical signs. Histopathological lesions included multifocal histiolymphocytic epicarditis, myocarditis and lymphocytic interstitial pneumonia, which are commonly seen in cases of MCF in buffaloes. Furthermore, lymphocytic vasculitis centered in the adventitia, with occasional fibrinoid necrosis in the muscular layer, was found in the kidneys, liver, spleen, lymph nodes and brain. Nucleotide sequencing of DNA fragments from the central nervous system amplified by PCR revealed 98% similarity with known OHV-2 sequences from Genbank. Additionally, PCR analysis also revealed the presence of OHV-2 DNA in the peripheral mononuclear blood cells of two clinically healthy buffaloes. The diagnosis of MCF was based on epidemiological, clinical, gross and histopathological findings and on the results of a semi-nested PCR followed by nucleotide sequencing.

Abstract in English:

ABSTRACT.- Mendes H.M.F., Faleiros R.R., Vasconcelos A.C., Alves G.E.S. & Moore R.M. 2009. [Apoptosis in equine small colon subjected to experimental ischemia and reperfusion.] Apoptose no cólon menor eqüino submetido à isquemia e reperfusão experimentais. Pesquisa Veterinária Brasileira 29(3):198-204. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: faleiros@ufmg.br Intestinal ischemia and reperfusion are important factors for mortality in horses. The objective of this study was to detect and to quantify apoptosis in the mucosa of equine small colon in a model of ischemia and reperfusion. The small colon was surgically exposed in twelve horses, and two intestinal segments were demarcated and subjected to 90 (SI) or 180 (SII) minutes of complete arteriovenous ischemia. Intestinal samples were collected before ischemia (control), at its end and after 90 and 180 minutes of reperfusion. Samples were histological processed and stained by hematoxylin and eosin (SI and SII) and by the technique of TUNEL (SI). Digitized histological images were analyzed morphometrically to detect apoptotic cells and to determine the apoptotic index (AI). After 90 or 180 minutes of arteriovenous ischemia, an increase in apoptotic cells was verified when compared with the control group, although no difference could be detected between the different periods of ischemia (P<0.05). After the first 90 minutes of reperfusion, a decrease in AI occurred, similar in both segments, possibly due to lack of energy source promoted by ischemia. AI was maximized after 180 minutes of reperfusion (sample harvested only in SI) (P<0.05). In conclusion, apoptosis is an important cause of cellular mucosal death in equine small colon ischemic obstruction, occurring early in ischemia, and later (after 90 minutes) in the reperfusion period.

• Horse small colon apoptosis ischemia and reperfusion

Abstract in Portuguese:

ABSTRACT.- Mendes H.M.F., Faleiros R.R., Vasconcelos A.C., Alves G.E.S. & Moore R.M. 2009. [Apoptosis in equine small colon subjected to experimental ischemia and reperfusion.] Apoptose no cólon menor eqüino submetido à isquemia e reperfusão experimentais. Pesquisa Veterinária Brasileira 29(3):198-204. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Pampulha, Belo Horizonte, MG 31270-901, Brazil. E-mail: faleiros@ufmg.br Intestinal ischemia and reperfusion are important factors for mortality in horses. The objective of this study was to detect and to quantify apoptosis in the mucosa of equine small colon in a model of ischemia and reperfusion. The small colon was surgically exposed in twelve horses, and two intestinal segments were demarcated and subjected to 90 (SI) or 180 (SII) minutes of complete arteriovenous ischemia. Intestinal samples were collected before ischemia (control), at its end and after 90 and 180 minutes of reperfusion. Samples were histological processed and stained by hematoxylin and eosin (SI and SII) and by the technique of TUNEL (SI). Digitized histological images were analyzed morphometrically to detect apoptotic cells and to determine the apoptotic index (AI). After 90 or 180 minutes of arteriovenous ischemia, an increase in apoptotic cells was verified when compared with the control group, although no difference could be detected between the different periods of ischemia (P<0.05). After the first 90 minutes of reperfusion, a decrease in AI occurred, similar in both segments, possibly due to lack of energy source promoted by ischemia. AI was maximized after 180 minutes of reperfusion (sample harvested only in SI) (P<0.05). In conclusion, apoptosis is an important cause of cellular mucosal death in equine small colon ischemic obstruction, occurring early in ischemia, and later (after 90 minutes) in the reperfusion period.

Abstract in English:

ABSTRACT.- Silva S.M.M.S., Castro R.S., Costa F.A.L., Vasconcelos A.C., Batista M.C.S., Riet-Correa F. & Carvalho E.M.S. 2007. [Epidemiology and symptoms of conidiobolomycosis in sheep in the State of Piauí, Brazil.] Epidemiologia e sinais clínicos da conidiobolomicose em ovinos no Estado do Piauí. Pesquisa Veterinária Brasileira 27(4):184-190. Departamento de Clínica e Cirurgia, Centro de Ciências Agrárias, Universidade Federal do Piauí, Campus da Socopo s/n, Teresina, PI 64049-550, Brazil. E-mail: silvanammss@ufpi.br Conidiobolomycosis is reported in 25 farms, from January 2002 to December 2004, in the state of Piauí. The disease affects only sheep, mainly in April-June. The mean morbidity rate was 2.80%, but was higher in the first semester (2.1%), during the raining period, than in the second one (0.69%), during the dry period. Morbidity rate among flocks varied from 0.1-14.3%. Case fatality rate was 100%, and the clinical manifestation period varied from 1-5 weeks. Clinical signs were serous, mucous and/or bloody nasal secretion, respiratory distress, snoring respiration, cranium-facial asymmetry, exophthalmia, fever and progressive emaciation. Marked depression, sometimes with the head down or head pressing was observed in some cases. Gross, microscopic and ultrastructural lesions and identification of the agent are reported elsewhere. This is the first report of conidiobolomycosis in Brazil, which is endemic and has a high frequency in sheep in the State of Piauí, associated with high rainfalls (1000-1600mm annually) and high temperature (19-36°C).

• Conidiobolomycosis Conidiobolus coronatus zygomycosis clinical signs epidemiology sheep

Abstract in Portuguese:

ABSTRACT.- Silva S.M.M.S., Castro R.S., Costa F.A.L., Vasconcelos A.C., Batista M.C.S., Riet-Correa F. & Carvalho E.M.S. 2007. [Epidemiology and symptoms of conidiobolomycosis in sheep in the State of Piauí, Brazil.] Epidemiologia e sinais clínicos da conidiobolomicose em ovinos no Estado do Piauí. Pesquisa Veterinária Brasileira 27(4):184-190. Departamento de Clínica e Cirurgia, Centro de Ciências Agrárias, Universidade Federal do Piauí, Campus da Socopo s/n, Teresina, PI 64049-550, Brazil. E-mail: silvanammss@ufpi.br Conidiobolomycosis is reported in 25 farms, from January 2002 to December 2004, in the state of Piauí. The disease affects only sheep, mainly in April-June. The mean morbidity rate was 2.80%, but was higher in the first semester (2.1%), during the raining period, than in the second one (0.69%), during the dry period. Morbidity rate among flocks varied from 0.1-14.3%. Case fatality rate was 100%, and the clinical manifestation period varied from 1-5 weeks. Clinical signs were serous, mucous and/or bloody nasal secretion, respiratory distress, snoring respiration, cranium-facial asymmetry, exophthalmia, fever and progressive emaciation. Marked depression, sometimes with the head down or head pressing was observed in some cases. Gross, microscopic and ultrastructural lesions and identification of the agent are reported elsewhere. This is the first report of conidiobolomycosis in Brazil, which is endemic and has a high frequency in sheep in the State of Piauí, associated with high rainfalls (1000-1600mm annually) and high temperature (19-36°C).