Ativação de células de memória na produção de anticorpos e na expressão de células IgM positivas no baço de tilápias-do-nilo / Activation of memory cells in the antibody production and the expression of IgM positive cells in Nile tilapia spleen

O presente trabalho avaliou o papel do baço no armazenamento e na reativação das linhagens de células B, representadas por células IgM positivas imunomarcadas no tecido esplênico, bem como a funcionalidade dessas células, sobre a cinética dos linfócitos e na produção sistêmica de anticorpos em tilápias-do-nilo (Oreochromis niloticus). Foram separados dois grupos: grupo memória, constituído por peixes previamente imunizados com hemácia de carneiro a 2,5%, para a geração da memória imune, e o grupo naive, que recebeu o mesmo volume de solução salina a 0,65%. Após 32 dias, os dois grupos foram submetidos a uma nova dose do antígeno na mesma concentração, volume e via de inoculação. A reativação dos clones de memória foi evidenciada pelo aumento do número de células IgM positivas no baço do grupo memória no dia zero/pré-imune. Além disso, o mesmo grupo apresentou aumento dos títulos de anticorpos séricos no 14º dia e no número absoluto de linfócitos no 21º dia em relação ao grupo naive. Esses resultados sugerem que o baço não seja apenas um local de armazenamento, mas também de reativação de células B de memória em tilápia-do-nilo.(AU)

This work aimed to evaluate the role of the spleen in storage and reactivation of the memory B cells, represented by IgM positive cells and the systemic production of sheep antibodies anti-red cell in Nile tilapia (Oreochromis niloticus). Two groups were established: the memory group, containing fish previously immunized with a 2,5% sheep anti-red cell, to generate the immune memory; and the naive group, containing fish that received a 0,65% saline solution. After 32 days, both groups were subjected to a new dose of the same antigen at the same concentration, volume, and inoculation via. The memory clones reactivation was correlated to the increase of the IgM positive cells in the spleen in the memory group at 0 day. The memory group showed an increase in the absolute number of lymphocytes at 21 days and an increase in the antibodies at 14 days after inoculation when compared to the naive group. The results suggest that the spleen may be a storage and reactivation place of memory B cells in Nile tilapia.(AU)

Ativação de células de memória na produção de anticorpos e na expressão de células IgM positivas no baço de tilápias-do-nilo / Activation of memory cells in the antibody production and the expression of IgM positive cells in Nile tilapia spleen

O presente trabalho avaliou o papel do baço no armazenamento e na reativação das linhagens de células B, representadas por células IgM positivas imunomarcadas no tecido esplênico, bem como a funcionalidade dessas células, sobre a cinética dos linfócitos e na produção sistêmica de anticorpos em tilápias-do-nilo (Oreochromis niloticus). Foram separados dois grupos: grupo memória, constituído por peixes previamente imunizados com hemácia de carneiro a 2,5%, para a geração da memória imune, e o grupo naive, que recebeu o mesmo volume de solução salina a 0,65%. Após 32 dias, os dois grupos foram submetidos a uma nova dose do antígeno na mesma concentração, volume e via de inoculação. A reativação dos clones de memória foi evidenciada pelo aumento do número de células IgM positivas no baço do grupo memória no dia zero/pré-imune. Além disso, o mesmo grupo apresentou aumento dos títulos de anticorpos séricos no 14º dia e no número absoluto de linfócitos no 21º dia em relação ao grupo naive. Esses resultados sugerem que o baço não seja apenas um local de armazenamento, mas também de reativação de células B de memória em tilápia-do-nilo.(AU)

This work aimed to evaluate the role of the spleen in storage and reactivation of the memory B cells, represented by IgM positive cells and the systemic production of sheep antibodies anti-red cell in Nile tilapia (Oreochromis niloticus). Two groups were established: the memory group, containing fish previously immunized with a 2,5% sheep anti-red cell, to generate the immune memory; and the naive group, containing fish that received a 0,65% saline solution. After 32 days, both groups were subjected to a new dose of the same antigen at the same concentration, volume, and inoculation via. The memory clones reactivation was correlated to the increase of the IgM positive cells in the spleen in the memory group at 0 day. The memory group showed an increase in the absolute number of lymphocytes at 21 days and an increase in the antibodies at 14 days after inoculation when compared to the naive group. The results suggest that the spleen may be a storage and reactivation place of memory B cells in Nile tilapia.(AU)

Deformities in reared cobia, Rachycentron canadum L. and grouper, Epinephelus marginatus, in São Paulo state coast, Brazil: case report / Deformidades em cobias, Rachycentron canadum L, e garoupas, Epinephelus marginatus, cultivadas na costa do estado de São Paulo, Brasil: relato de caso

Skeletal deformities are frequent problems in cultured fish populations due mainly to unfavorable abiotic conditions, inappropriate nutrition, and genetic factors. These may cause erratic swimming, decrease of conversion rate, growth, and market value. The aim of this case report was to present the occurrence of deformities in juvenile cobia and groupers from an offshore marine farm in Ilhabela, São Paulo, Brazil. Nine cobias and seven juvenile groupers, that presented deformities, were euthanized and fixed in 10% formaldehyde. They were tagged, processed, and stained with "Alcian Blue" (AB) and "Alizarin Red-S" (ARS) for visualization of cartilage and bone tissue, respectively. After evisceration, radiographic examination was performed. The deformities observed in cobia were atrophy of the operculum, disorganization of the gill lamellae, deformity of the lower jaw, kyphosis, and scoliosis. Groupers presented lower jaw deformity and saddleback syndrome. The farm studied in this case report had poor management standards, fed commercial feed for freshwater herbivorous fish and cooked sardines, high inbreeding and absence of adequately trained professionals to manage these species. The sum of these factors must have directly influenced the appearance of these deformities. It is necessary to control this situation to minimize the incidence of malformations that have economic impacts to farmers.(AU)

As deformidades esqueléticas são problemas frequentes em populações de peixes cultivados devido, principalmente, a condições abióticas desfavoráveis, nutrição inapropriada e fatores genéticos. Estas podem causar natação errática, diminuição do índice de conversão, do crescimento e do valor no mercado. O objetivo deste relato de caso foi apresentar a ocorrência de deformidades em cobias e garoupas juvenis provenientes de uma fazenda marinha em sistema offshore, em Ilhabela, São Paulo, Brasil. Nove cobias e sete garoupas juvenis, que apresentavam deformidades, foram sujeitos à eutanásia e fixados em formol a 10%. Os espécimes foram registrados, processados e corados com "Alcian Blue" (AB) e "Alizarin Red-S" (ARS) para visualização de tecido cartilaginoso e ósseo, respectivamente. Os peixes foram eviscerados para o exame radiográfico. As deformidades observadas em cobia foram: atrofia do opérculo, desorganização das lamelas branquiais, deformidade do maxilar inferior, cifose e escoliose. As garoupas apresentaram deformidade do maxilar inferior e síndrome de Saddleback. Os peixes deste relato de caso estiveram sob manejo deficiente, alimentados com ração comercial para peixes herbívoros de água doce e sardinhas cozinhadas, alta consanguinidade e ausência de profissionais adequadamente treinados para o manejo dessas espécies. A soma desses fatores influenciou diretamente na aparição dessas deformidades, precisando ser controlados para minimizar a incidência de malformações que tenham impacto econômico para os produtores.(AU)

Deformities in reared cobia, Rachycentron canadum L. and grouper, Epinephelus marginatus, in São Paulo state coast, Brazil: case report / Deformidades em cobias, Rachycentron canadum L, e garoupas, Epinephelus marginatus, cultivadas na costa do estado de São Paulo, Brasil: relato de caso

Skeletal deformities are frequent problems in cultured fish populations due mainly to unfavorable abiotic conditions, inappropriate nutrition, and genetic factors. These may cause erratic swimming, decrease of conversion rate, growth, and market value. The aim of this case report was to present the occurrence of deformities in juvenile cobia and groupers from an offshore marine farm in Ilhabela, São Paulo, Brazil. Nine cobias and seven juvenile groupers, that presented deformities, were euthanized and fixed in 10% formaldehyde. They were tagged, processed, and stained with "Alcian Blue" (AB) and "Alizarin Red-S" (ARS) for visualization of cartilage and bone tissue, respectively. After evisceration, radiographic examination was performed. The deformities observed in cobia were atrophy of the operculum, disorganization of the gill lamellae, deformity of the lower jaw, kyphosis, and scoliosis. Groupers presented lower jaw deformity and saddleback syndrome. The farm studied in this case report had poor management standards, fed commercial feed for freshwater herbivorous fish and cooked sardines, high inbreeding and absence of adequately trained professionals to manage these species. The sum of these factors must have directly influenced the appearance of these deformities. It is necessary to control this situation to minimize the incidence of malformations that have economic impacts to farmers.(AU)

As deformidades esqueléticas são problemas frequentes em populações de peixes cultivados devido, principalmente, a condições abióticas desfavoráveis, nutrição inapropriada e fatores genéticos. Estas podem causar natação errática, diminuição do índice de conversão, do crescimento e do valor no mercado. O objetivo deste relato de caso foi apresentar a ocorrência de deformidades em cobias e garoupas juvenis provenientes de uma fazenda marinha em sistema offshore, em Ilhabela, São Paulo, Brasil. Nove cobias e sete garoupas juvenis, que apresentavam deformidades, foram sujeitos à eutanásia e fixados em formol a 10%. Os espécimes foram registrados, processados e corados com "Alcian Blue" (AB) e "Alizarin Red-S" (ARS) para visualização de tecido cartilaginoso e ósseo, respectivamente. Os peixes foram eviscerados para o exame radiográfico. As deformidades observadas em cobia foram: atrofia do opérculo, desorganização das lamelas branquiais, deformidade do maxilar inferior, cifose e escoliose. As garoupas apresentaram deformidade do maxilar inferior e síndrome de Saddleback. Os peixes deste relato de caso estiveram sob manejo deficiente, alimentados com ração comercial para peixes herbívoros de água doce e sardinhas cozinhadas, alta consanguinidade e ausência de profissionais adequadamente treinados para o manejo dessas espécies. A soma desses fatores influenciou diretamente na aparição dessas deformidades, precisando ser controlados para minimizar a incidência de malformações que tenham impacto econômico para os produtores.(AU)

Transferência passiva de soro hiperimune anti-Streptococcus agalactiae e seu efeito profilático em tilápias-do-nilo infectadas experimentalmente: sobrevivência e títulos de anticorpos / Passive transfer of hyperimmune serum anti Streptococcus agalactiae and its prophylactic effect on Nile tilapia experimentally infected

A bactéria Streptococcus agalactiae é um potente agente causador de surtos por doenças bacterianas em peixes. O estresse provocado pelo manejo zootécnico e pela má qualidade ambiental torna a tilápia susceptível às infecções por essa bactéria. O objetivo do presente trabalho foi avaliar a resistência de tilápias-do-nilo imunizadas com soro hiperimune anti-S. agalactiae, posteriormente desafiadas com cepa homóloga da mesma bactéria. Após determinação da DL 50 de S. agalactiae, 36 tilápias foram distribuídas em quatro aquários, dois para o grupo controle e dois para inoculação celomática para produção de anticorpos anti-S. agalactiae. No 21° e 28° dias, foi coletado sangue para obtenção de soro hiperimune utilizado na transferência passiva. Em seguida, 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos: GI: controle; GII: imunizadas com o soro inativado; GIII: imunizadas com soro ativo. Após 48 horas e sete, 14, 21, 28 e 35 dias, foram realizadas coletas de sangue para titulação de anticorpos anti-S. agalactiae utilizando-se o teste de aglutinação direta. Para avaliar a taxa de sobrevivência, outras 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos (GI: controle; GII: imunizadas com soro inativado; GIII: imunizadas com soro ativo). Após 48 horas da imunização, as tilápias foram desafiadas via celomática com 100µL de S. agalactiae e avaliadas duas vezes ao dia, pelo período de 35 dias. Os resultados dos títulos séricos de anticorpos foram detectados pela aglutinação direta até o 21° dia pós-transferência passiva, e, no mesmo período, houve proteção de 80% entre os grupos imunizados com soro inativado e soro ativo contendo anticorpos anti-S. agalactiae. Ao final, os grupos soro inativado e soro ativo apresentaram 60 e 80% de proteção, respectivamente, enquanto no grupo controle 100% dos peixes adoeceram, apresentando sinais graves da infecção, e foram eutanasiados. Não houve diferença estatística significativa na taxa de proteção entre os grupos imunizados.(AU)

The Streptococcus agalactiae bacteria is a potent agent which causes outbreaks of bacterial diseases in fish. The stress caused by management and poor environmental quality makes tilapia susceptible to infections, including by bacterium. The aim of this study was to evaluate the resistance of the Nile tilapia immunized with hyperimmune serum against S. agalactiae subsequently challenged with homologous strain of the same bacteria. After determining the DL 50 of S. agalactiae, 36 tilapias were distributed in 4 aquariums, 2 for the control group and 2 for the group via coelomic, inoculated with the DL 50 for anti-S. agalactiae antibodies production. On the 21st and 28th day blood was collected for the obtainment of hiperimmune serum used in passive transference. Then, 30 tilapias were distributed in 3 aquariums submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum). After 48 hours, 7, 14, 21, 28 and 35 days blood collections were done for anti-S. agalactiae antibodies titration using the direct agglutination test. For survival evaluation 30 tilapias were distributed in 3 aquariums and submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum). After 48 hours of inoculation tilapias were challenged with 100µL of S. agalactiae and monitored twice a day for 35 days. The results showed that titers of antibodies were detected by direct agglutination until the 21st day after passive transference and during the same period the protection between the groups immunized with inactivated serum and non-inactivated serum contending anti-S. agalactiae antibodies was of 80% after challenge with S. agalactiae. In the end, the serum-inactivated serum and active groups had 60 and 80% protection, respectively, while 100% of the fish in the control group developed severe signs of infection and were euthanized. There was no statistical difference in the survival rate between the immunized groups.(AU)

Transferência passiva de soro hiperimune anti-Streptococcus agalactiae e seu efeito profilático em tilápias-do-nilo infectadas experimentalmente: sobrevivência e títulos de anticorpos / Passive transfer of hyperimmune serum anti Streptococcus agalactiae and its prophylactic effect on Nile tilapia experimentally infected

A bactéria Streptococcus agalactiae é um potente agente causador de surtos por doenças bacterianas em peixes. O estresse provocado pelo manejo zootécnico e pela má qualidade ambiental torna a tilápia susceptível às infecções por essa bactéria. O objetivo do presente trabalho foi avaliar a resistência de tilápias-do-nilo imunizadas com soro hiperimune anti-S. agalactiae, posteriormente desafiadas com cepa homóloga da mesma bactéria. Após determinação da DL 50 de S. agalactiae, 36 tilápias foram distribuídas em quatro aquários, dois para o grupo controle e dois para inoculação celomática para produção de anticorpos anti-S. agalactiae. No 21° e 28° dias, foi coletado sangue para obtenção de soro hiperimune utilizado na transferência passiva. Em seguida, 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos: GI: controle; GII: imunizadas com o soro inativado; GIII: imunizadas com soro ativo...

The Streptococcus agalactiae bacteria is a potent agent which causes outbreaks of bacterial diseases in fish. The stress caused by management and poor environmental quality makes tilapia susceptible to infections, including by bacterium. The aim of this study was to evaluate the resistance of the Nile tilapia immunized with hyperimmune serum against S. agalactiae subsequently challenged with homologous strain of the same bacteria. After determining the DL 50 of S. agalactiae, 36 tilapias were distributed in 4 aquariums, 2 for the control group and 2 for the group via coelomic, inoculated with the DL 50 for anti-S. agalactiae antibodies production. On the 21st and 28th day blood was collected for the obtainment of hiperimmune serum used in passive transference. Then, 30 tilapias were distributed in 3 aquariums submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum)...

Muscular dystrophy in dogs: does the crossing of breeds influence disease phenotype?

Golden Retriever (GR) muscular dystrophy is an inherited degenerative muscle disease that provides an excellent model for Duchenne muscular dystrophy in humans. This study defined the histopathologic lesions, including the distribution of type I and II muscle fibers (FTI and FTII), in 12 dystrophic and 3 nondystrophic dogs between 7 and 15 months of age. The authors were interested in studying the influence on disease phenotype from crossing the base GR breed with Yellow Labrador Retrievers. The dystrophic dogs were divided according to breed: GRs and Golden Labrador Retrievers (GLRs). On hematoxylin and eosin staining, histopathologic lesions were more severe in GRs than GLRs. Six of eight GR muscles (75%) had a severe lesion grade (grade 3). In contrast, seven GLR muscles (87.5%) had mild lesions (grade 2), and only one had severe lesions (grade 3). Changes in fiber-type distribution were more pronounced in GRs versus GLRs. FTI:FTII ratio inversion was observed in three dystrophic GRs but only one GLR. The mean diameter of FTI and FTII was smaller in GRs and GLRs than in nondystrophic dogs (P < .01). The FTI of five GR muscles (62.5%) were larger than those of GLRs, whereas only one GLR muscle was larger (P < .05). The differential was less pronounced for FTII, with four GR muscles being larger and three GLR being larger. Observations indicate that crossing the base GR breed with Labrador Retrievers lessened the severity of the GR muscular dystrophy phenotype.

Effects of glycosaminoglycan polysulphate on the organisation of collagen fibres in experimentally induced tendonitis in horses.

An inflammatory process was induced by intratendinous injection of bacterial collagenase into the superficial digital flexor tendon (SDFT) of the left thoracic limb of 10 horses. One week later, the tendons in five of the horses (group 1) were treated with glycosaminoglycan polysulphate (GAGPS), and the tendons of the other five (group 2) were treated with saline solution. The horses were euthanased 150 days after the collagenase injections, and samples of the SDFTs were frozen at -14 degrees C, sectioned at 5 to 7 mum longitudinally and transversely, and stained by the picrosirius red method. Morphometric analysis was used to quantify the organised and disorganised bundles of collagen in the samples from groups 1 and 2. Significantly more organised bundles of collagen were observed in the tendons treated with GAGPS.

Renal involvement in canine leishmaniasis: a morphological and immunohistochemical study / Envolvimento renal na leishmaniose canina: estudo morfológico e imunoistoquímico

IgG and IgM deposits in kidneys of dogs with visceral leishmaniasis (VL) were studied in 25 symptomatic dogs (case) and 15 asymptomatic dogs (control) by an immunohistochemical method. All tested dogs were positive for VL by polymerase chain reaction, enzyme-linked immunosorbent assay, and indirect immunofluorescence test. Kidney fragments were submitted to immunohistochemical reaction. Many morphological patterns of distribution of subendothelial granules were identified for IgG and IgM in glomerular capillaries: global, segmental, diffuse, or focal. Intensity of immunohistochemical reaction to IgG was not significantly different when comparing the symptomatic and the asymptomatic animal groups by Fisher's exact test. IgM reactions were significantly different between groups (P<0.01). Deposits of IgM on mesangial cells and in inflammatory interstitial infiltrate were rarely seen, although IgG reactions were frequent at these sites. This study concluded that immunohistochemical reactions for IgM were more intense than those observed for IgG in canine VL, and these reactions were characterized by distribution of subendothelial granules in glomerular capillaries.(AU)

Caracterizou-se a deposição de IgG e IgM em rins de cães com leishmaniose visceral (LV) pelo uso da técnica imunoistoquímica. Foram estudados rins de 25 cães sintomáticos (caso) e de 15 cães assintomáticos (controle). Todos os animais foram positivos para leishmaniose pela reação em cadeia da polimerase, pelo ELISA e pela imunofluorescência indireta. Fragmentos renais foram submetidos à reação imunoistoquímica. Diversos padrões morfológicos de distribuição de grânulos subendoteliais de IgG e IgM foram identificados nos capilares glomerulares: global, segmentar, difuso ou focal. A intensidade da reação imunoistoquímica da IgG, medida pelo teste exato de Fisher não diferiu entre os grupos sintomáticos e assintomáticos e a intensidade de reação da IgM foi diferente entre os grupos (P<0,01). Depósitos de IgM nas células mesangiais e no infiltrado inflamatório raramente foram visualizados, no entanto as reações IgG foram freqüentemente visualizadas nesses locais. Concluiu-se que as reações de IgM foram mais intensas que as reações de IgG na LV canina e caracterizam-se pela distribuição de grânulos subendoteliais nos capilares glomerulares.(AU)

Renal involvement in canine leishmaniasis: a morphological and immunohistochemical study / Envolvimento renal na leishmaniose canina: estudo morfológico e imunoistoquímico

IgG and IgM deposits in kidneys of dogs with visceral leishmaniasis (VL) were studied in 25 symptomatic dogs (case) and 15 asymptomatic dogs (control) by an immunohistochemical method. All tested dogs were positive for VL by polymerase chain reaction, enzyme-linked immunosorbent assay, and indirect immunofluorescence test. Kidney fragments were submitted to immunohistochemical reaction. Many morphological patterns of distribution of subendothelial granules were identified for IgG and IgM in glomerular capillaries: global, segmental, diffuse, or focal. Intensity of immunohistochemical reaction to IgG was not significantly different when comparing the symptomatic and the asymptomatic animal groups by Fisher's exact test. IgM reactions were significantly different between groups (P<0.01). Deposits of IgM on mesangial cells and in inflammatory interstitial infiltrate were rarely seen, although IgG reactions were frequent at these sites. This study concluded that immunohistochemical reactions for IgM were more intense than those observed for IgG in canine VL, and these reactions were characterized by distribution of subendothelial granules in glomerular capillaries.

Caracterizou-se a deposição de IgG e IgM em rins de cães com leishmaniose visceral (LV) pelo uso da técnica imunoistoquímica. Foram estudados rins de 25 cães sintomáticos (caso) e de 15 cães assintomáticos (controle). Todos os animais foram positivos para leishmaniose pela reação em cadeia da polimerase, pelo ELISA e pela imunofluorescência indireta. Fragmentos renais foram submetidos à reação imunoistoquímica. Diversos padrões morfológicos de distribuição de grânulos subendoteliais de IgG e IgM foram identificados nos capilares glomerulares: global, segmentar, difuso ou focal. A intensidade da reação imunoistoquímica da IgG, medida pelo teste exato de Fisher não diferiu entre os grupos sintomáticos e assintomáticos e a intensidade de reação da IgM foi diferente entre os grupos (P<0,01). Depósitos de IgM nas células mesangiais e no infiltrado inflamatório raramente foram visualizados, no entanto as reações IgG foram freqüentemente visualizadas nesses locais. Concluiu-se que as reações de IgM foram mais intensas que as reações de IgG na LV canina e caracterizam-se pela distribuição de grânulos subendoteliais nos capilares glomerulares.

Influência da heparina sódica na ocorrência de laminite eqüina induzida por sobrecarga de carboidratos / Influence of heparin in occurrence of carbohydrate overload-induced equine laminitis

Avaliou-se a eficiência da infusão intravenosa de heparina sódica (100UI/kg/8h, a partir de 24h após o fornecimento de carboidrato, até completar 48h) no controle da laminite eqüina experimentalmente induzida por sobrecarga de carboidrato (17,6g de amido de milho/kg de peso corpóreo). Foram utilizados 15 eqüinos adultos, distribuídos em três grupos experimentais: GI (grupo-controle); GII (grupo laminite) e GIII (grupo laminite+heparina). Posteriormente ao fornecimento de carboidrato, os animais foram submetidos a exames físicos e laboratoriais durante um período de 48 horas. Ao final do experimento, os animais foram submetidos à eutanásia pela aplicação intravenosa de 5ml de maleato de acepromazina seguida de 1g de tiopental sódico e 1 litro de solução saturada de KCl para a obtenção de amostras de tecidos dos cascos, necessárias ao exame histológico. Os animais de GII e GIII, submetidos à sobrecarga de carboidratos, desenvolveram laminite, exibindo claudicação 12 e 24h após o fornecimento de carboidrato, respectivamente, bem como aumentos da freqüência cardíaca e do tempo de preenchimento capilar. As alterações histológicas, semelhantes em GII e GIII, eram do tipo degenerativo, como adelgaçamento de lâminas epidérmicas, retração, achatamento e deslocamento de lâminas dérmicas, vacuolização epidérmica e desorganização do tecido epidérmico. A infusão da heparina sódica não preveniu ou atenuou a degeneração laminar.(AU)

The efficacy of intravenous heparin administration (100UI/kg/8h, from 24 to 48h after carbohydrate administration) in the control of carbohydrate overload-induced equine laminitis (17.6g of corn starch/kg live weight) was evaluated. Fifteen horses were allocated into three experimental groups: GI (control group), GII (laminitis group), and GIII (laminitis+heparin group). These animals were submitted to physical and laboratorial examination during 48h. After that time, they were euthanized with intravenous administration of 5ml of acepromazine followed by 1g of thiopental sodium and 1 liter of saturated solution of KCl to obtain hoof tissues samples for histological examination. GII and GIII horses developed laminitis, showing lameness 12 and 24h after carbohydrate administration, respectively, as well increased heart rate and capillary refill time. The histological alterations, similar in GII and GIII, were degenerative lesions, as thinness of epidermal lamina, retraction, flattening and dislocation of the dermal lamina, epidermal vacuolization, and disruption of the epidermal tissues. The occurrence of laminar degeneration was not prevented or attenuated with intravenous heparin administration.(AU)

Influência da heparina sódica na ocorrência de laminite eqüina induzida por sobrecarga de carboidratos / Influence of heparin in occurrence of carbohydrate overload-induced equine laminitis

Avaliou-se a eficiência da infusão intravenosa de heparina sódica (100UI/kg/8h, a partir de 24h após o fornecimento de carboidrato, até completar 48h) no controle da laminite eqüina experimentalmente induzida por sobrecarga de carboidrato (17,6g de amido de milho/kg de peso corpóreo). Foram utilizados 15 eqüinos adultos, distribuídos em três grupos experimentais: GI (grupo-controle); GII (grupo laminite) e GIII (grupo laminite+heparina). Posteriormente ao fornecimento de carboidrato, os animais foram submetidos a exames físicos e laboratoriais durante um período de 48 horas. Ao final do experimento, os animais foram submetidos à eutanásia pela aplicação intravenosa de 5ml de maleato de acepromazina seguida de 1g de tiopental sódico e 1 litro de solução saturada de KCl para a obtenção de amostras de tecidos dos cascos, necessárias ao exame histológico. Os animais de GII e GIII, submetidos à sobrecarga de carboidratos, desenvolveram laminite, exibindo claudicação 12 e 24h após o fornecimento de carboidrato, respectivamente, bem como aumentos da freqüência cardíaca e do tempo de preenchimento capilar. As alterações histológicas, semelhantes em GII e GIII, eram do tipo degenerativo, como adelgaçamento de lâminas epidérmicas, retração, achatamento e deslocamento de lâminas dérmicas, vacuolização epidérmica e desorganização do tecido epidérmico. A infusão da heparina sódica não preveniu ou atenuou a degeneração laminar.

The efficacy of intravenous heparin administration (100UI/kg/8h, from 24 to 48h after carbohydrate administration) in the control of carbohydrate overload-induced equine laminitis (17.6g of corn starch/kg live weight) was evaluated. Fifteen horses were allocated into three experimental groups: GI (control group), GII (laminitis group), and GIII (laminitis+heparin group). These animals were submitted to physical and laboratorial examination during 48h. After that time, they were euthanized with intravenous administration of 5ml of acepromazine followed by 1g of thiopental sodium and 1 liter of saturated solution of KCl to obtain hoof tissues samples for histological examination. GII and GIII horses developed laminitis, showing lameness 12 and 24h after carbohydrate administration, respectively, as well increased heart rate and capillary refill time. The histological alterations, similar in GII and GIII, were degenerative lesions, as thinness of epidermal lamina, retraction, flattening and dislocation of the dermal lamina, epidermal vacuolization, and disruption of the epidermal tissues. The occurrence of laminar degeneration was not prevented or attenuated with intravenous heparin administration.

CLINICAL AND LABORATORIAL EVALUATION IN PRODROMAL STAGE OF CARBOHYDRATE OVERLOAD-INDUCED EQUINE LAMINITIS. / ESTUDO CLÍNICO E LABORATORIAL DA FASE PRODRÔMICA DA LAMINITE EQÜINA INDUZIDA POR SOBRECARGA DE CARBOIDRATO.

Laminitis is a complex disorder that is difficult to diagnose before the onset of lameness. Ten horses were allocated into 2 experimental groups: The 5 animals of GI did not undergo carbohydrate overload (control group) whereas the remaining 5 in GII underwent carbohydrate overload (laminitis group). After carbohydrate was provided to the animals, clinical and laboratorial examination were performed over a 48-hour period. The hoof tissues underwent histological assessment at the end of study. The digital pulse increased from 12 hours on, simultaneous to initial signs of lameness, which increased to Obel grade 3 after 24 hours. Change of external maxilar artery pulse was noted to 24 hours; at the same time an increase of the capillary refill time was seen. It was verified that heart rate and internal temperature increased 36 hours after overfeeding. Eosinopenia occurred 12 hours after carbohydrate overload. The packet cell volume changed after 36 hours, concomitantly with neutrophilia. Mild leucocytosis occurred during the experiment. The changes of the plasma total protein and fibrinogen concentrations and serum aspartate aminotransferase activity were not significant. Although not expected, the serum creatine kynase activity was highest in control group. The serum alkaline phosphatase activity was highest in laminitis group. The major histological changes were degenerative, in

Laminite é um complexo distúrbio metabólico de difícil diagnostico antes do início da claudicação. Foram utilizados dez eqüinos adultos distribuídos em 2 grupos experimentais: GI, 5 animais hígidos (grupo controle) e GII, 5 animais submetidos à dieta com sobrecarga de carboidratos (grupo laminite). Posteriormente ao fornecimento de carboidratos, os animais foram submetidos a exames físicos e laboratorial durante o período de 48 horas. Ao final do experimento procedeu-se a avaliação histológica dos tecidos dos cascos. Notou-se que o pulso digital elevou-se a partir de 12 horas após o início da fase experimental, simultaneamente aos primeiros sinais de claudicação, que evoluíram para grau III 24 horas após. Alteração do pulso da artéria maxilar externa foi notada a partir de 24 horas, coincidindo com o aumento do tempo de preenchimento capilar. Elevações de freqüência cardíaca e da temperatura retal ocorreram 36 a 48 horas após o fornecimento de carboidratos. Notou-se eosinopenia 12 horas após a administração de carboidratos. O volume globular alterou-se a partir de 36 horas, concomitantemente à neutrofilia. Constatou-se discreta leucocitose ao longo da fase experimental. As alterações dos teores plasmáticos de proteína total e de fibrinogênio e a atividade sérica de aspartato aminotransferase não foram significativas. A atividade sérica de creatina quinase foi maior em eqüinos d

Alterações morfológicas e funcionais dos rins de cães com insuficiência renal crônica / Morphologic and functional alterations of the kidneys of dogs with chronic renal failure

Alterações morfológicas de 11 casos de cães com insuficiência renal foram caracterizadas e classificadas de acordo com os padrões estabelecidos pela Organização Mundial de Saúde para seres humanos. Glomerulonefrite esclerosante difusa foi diagnosticada em 82,0% dos animais e nefrite intersticial crônica nos 18,0% restantes. Os tipos e freqüência das lesões identificadas foram similares às encontradas na literatura para a insuficiência renal crônica.(AU)

Morphologic alterations of 11 cases of dogs with renal failure were characterized and classified according to the patterns established by the World Health Organization for human beings. Diffuse sclerosing glomerulonephritis was diagnosed in 82.0% of the animals and chronic interstitial nephritis in the remaining 18.0%. The types and frequencies of lesions were similar to the those noticed in the literature for chronic renal failure.(AU)

Alterações morfológicas e funcionais dos rins de cães com insuficiência renal crônica / Morphologic and functional alterations of the kidneys of dogs with chronic renal failure

Alterações morfológicas de 11 casos de cães com insuficiência renal foram caracterizadas e classificadas de acordo com os padrões estabelecidos pela Organização Mundial de Saúde para seres humanos. Glomerulonefrite esclerosante difusa foi diagnosticada em 82,0 por cento dos animais e nefrite intersticial crônica nos 18,0 por cento restantes. Os tipos e freqüência das lesões identificadas foram similares às encontradas na literatura para a insuficiência renal crônica.

Morphologic alterations of 11 cases of dogs with renal failure were characterized and classified according to the patterns established by the World Health Organization for human beings. Diffuse sclerosing glomerulonephritis was diagnosed in 82.0 percent of the animals and chronic interstitial nephritis in the remaining 18.0 percent. The types and frequencies of lesions were similar to the those noticed in the literature for chronic renal failure.

Clinical and morphological evaluation of snake venom derived fibrin glue on the tendon healing in dogs

The aim of this study was to evaluate the effect of snake venom derived fibrin glue on the healing of the deep digital flexor tendon, during three periods. The tendon of the 2nd digit of 30 thoracic limbs of dogs was partially sectioned for glue application. Biopsies were performed 7, 15, and 30 days post surgery for the clinical and morphological study of tendons. Analysis of the results showed that 73.3% of the tendons showed stump retraction and 16.6% moderate to excessive adherence, which affected sliding. There was a significant difference in the number of inflammatory cells among the three studied periods, being the highest on day 15. The morphological analysis revealed a typical tendon healing process with a lower level of inflammation in the acute phase, facilitating the cicatricial maturation phase. Snake venom derived fibrin glue promotes the healing in dog flexor tendon

Polymerase chain reaction in detecting Leishmania sp in symptomatic and asymptomatic seropositive dogs

In human and canine renal histological studies of visceral leishmaniasis (VL), the etiological agent is rarely found in situ. The objective of this study was to evaluate PCR in identifying the etiological agent in spleen, liver, lymph node, and kidneys of VL-seropositive dogs. Twenty-five symptomatic (case group) and 15 asymptomatic (control group) VL-seropositive dogs of different breeds, sexes, and ages from Teresina, Piauí State, Brazil, were used. Serologic diagnosis was made by enzyme-linked immunosorbent assay and indirect immunofluorescence test. Animals were subjected to euthanasia and necropsy. Renal fragments were immersed in buffered formaldehyde solution. Spleen, liver, lymph node, and kidney samples were collected and frozen at -70ºC until DNA extraction. After dehydration and diaphanization, renal fragments were infiltrated and embedded in paraffin, cut at 3 mm, and stained with hematoxylin-eosin (HE). DNA amplification used an automatic thermocycler with specific Leishmania primers. All case-group dogs and 2 controls showed positive results in spleen, liver, or lymph node PCRs. There was a significant difference by Fisher exact test. In symptomatic seropositive dogs, renal histopathological evaluation showed one animal (4%) with amastigote forms of Leishmania in inflammatory infiltrate, and kidney PCRs detected Leishmania DNA in eight animals (32%). The conclusion was that PCR is more precise than the conventional histopathology in detecting the Leishmania parasite in kidney

Renal involvement in visceral leishmaniasis dogs

Visceral leishmaniasis (VL) is a zoonosis that affects both animals and man. Dogs are the etiological agent’s main reservoir. The aim of this study was to evaluate the clinical laboratory aspects and renal histopathology of VL dogs. Thirty-four symptomatic (case) and 17 asymptomatic (control) VL seropositive dogs of different breeds, sexes, and ages from Teresina, Piauí State, Brazil, were used. Diagnosis was confirmed by enzyme-linked immunosorbent assay and indirect immunofluorescence test. Clinical and laboratory tests included blood cell count and renal function analysis (urea and creatinine). Animals were subjected to euthanasia and necropsy. Renal fragments were prepared by the usual histological techniques and stained with hematoxylin-eosin and periodic acid-Schiff. Physical examination showed that lymph node hypertrophy (85.29%) and skin lesions (35.29%) were frequent in the case group. Anemia was found in 55.88% of the case and in 11.76% of the control group. There was a significant difference between groups by Fisher’s exact test. Two case-group dogs showed azotemia. Renal histopathological evaluation showed that 61.76% case and 17.65% control-group dogs had membranoproliferative glomerulonephritis. Mesangial proliferative glomerulonephritis was seen in 32.35% case and 64.70% control-group animals. There was a significant difference for both types of glomerulonephritis between groups. Amastigote forms of Leishmania were found in the renal parenchyma, in the inflammatory infiltrate of one case-group dog. We concluded that, in canine VL, regardless of the clinical signs at physical examination, the kidneys are frequently compromised

Effect of snake venom derived fibrin glue on the tendon healing in dogs. Clinical and biomechanical study

The aim of this study was to evaluate the effect of snake venom derived fibrin glue on the strength of tendon healing in dogs. The deep digital flexor tendon of the 5th digit of 24 thoracic limbs was partially sectioned for adhesive application. On the 7th, 15th, and 30th postoperative day tendons segments were removed for the clinical and biomechanical study. Results indicated that 62.5% of the tendons showed stump retraction and 20.8% moderate to excessive adherence, which affected gliding. The biomechanical evaluation showed that, over time, tendon healing gained progressive resistance for maximum traction and permanent deformations with satisfactory results on the 15th day for rigidity and resilience compared to the other two studied periods. Snake venom derived fibrin glue promotes healing in dog flexor tendon

Snake venom derived fibrin adhesive in dog uterine healing. Biomechanical study

Healing is a complex process with many interfering factors. The objective of this work was to evaluate regeneration strenght of non-pregnant adult dog uterus when snake venom derived fibrin adhesive is used to reinforce hysterorrhaphy. Maximum limit and rigidity were analyzed. Twenty uterine horns from 10 dogs were hysterotomized and distributed into 2 groups. Hysterorrhaphy was performed using the Shimieden-Cushing double layer suture. In one group, animals received snake venom derived fibirn adhesive as reinforcement. Although neither variable was significantly different, our results showed higher rigidity values in the adhesive group. This can be attributed to the adhesive's effect on organ elasticity or to more granulation tissue formed in the uterine scar