Zootechnical performance, degree of steatosis and the genotoxic potential in yellowtail tetra Astyanax lacustris fed with different levels of L-carnitine / Desempenho zootécnico, grau de esteatose e potencial genotóxico de lambaris Astyanax lacustris alimentados com diferentes níveis de L-carnitina

L-carnitine perform a major role in transporting long-chain fatty acids into the mitochondria, where they are oxidized. It has been used in animal diets to decrease fat and increase muscle protein. The aim of this study was to evaluate the zootechnical performance, degree of steatosis in the liver, and genotoxic potential in Astyanax lacustris fed with different levels of L-carnitine (LC). Yellowtail tetra juveniles (n = 140) were distributed in 20 tanks of 70 L, with seven fish in each, in a water recirculation system with controlled temperature (27±0.1°C). The treatments with different levels of L-carnitine supplementation were: 0 (control), 250, 500, 750, and 1000 mg of LC per kg of food. The diets were provided twice a day for 60 days. The results showed that the different levels of LC did not affect (P>0.05) weight gain, survival, viscerosomatic index, and the liver hepatocytes showed a normal appearance. However, the use of LC supplementation showed genotoxic potential with a significant difference (P<0.05) for cell alterations when compared to the control at concentrations above 500mg kg-1.

A L-carnitina exerce um papel importante no transporte de ácidos graxos de cadeia longa até a mitocôndria para serem oxidados e tem sido incorporada em rações para animais com o objetivo de diminuir a deposição de gordura e aumentar a proteína muscular. O objetivo deste trabalho foi avaliar o desempenho zootécnico, o grau de esteatose no fígado e o potencial genotóxico em Astyanax lacustris alimentados com diferentes níveis de L-carnitina (LC). Juvenis de lambari-do-rabo-amarelo (n=140) foram distribuídos em 20 caixas de 70L, sete peixes em cada, em um sistema de recirculação de água com temperatura controlada (27±0,1°C). Os tratamentos com os níveis de suplementação foram: 0 (controle), 250, 500, 750 e 1000 mg de LC kg-1 de ração. As dietas foram fornecidas duas vezes ao dia, durante 60 dias. Os resultados mostraram que os diferentes níveis de LC não influenciaram (P>0,05) o ganho de peso; a sobrevivência, o índice viscerossomático e os hepatócitos do fígado apresentaram-se com aparência normal. No entanto, a suplementação com LC apresentou potencial genotóxico com diferença significativa (P<0,05) para alterações celulares quando comparada ao controle em concentrações superiores a 500mg kg-1.

Antifungal activity of terpenes isolated from the Brazilian Caatinga: a review / Atividade antifúngica de terpenos isolados da Caatinga: uma revisão

Terpenoids, also named terpenes or isoprenoids, are a family of natural products found in all living organisms. Many plants produce terpenoids as secondary metabolites, and these make up a large part of essential oils. One of most important characteristic is that the compounds are volatile, have odor and can be used in a variety of applications in different industrial segments and traditional medicine. Brazil has a rich and diverse flora that can be used as a source of research for obtaining new molecules. Within the Brazilian flora, it is worth mentioning the Caatinga as an exclusively Brazilian biome where plants adapt to a specific series of weather conditions and therefore become a great storehouse of the terpenoid compounds to be described herein. Fungal infections have become increasingly common, and a great demand for new agents with low toxicity and side effects has thus emerged. Scientists must search for new molecules exhibiting antifungal activity to develop new drugs. This review aims to analyze scientific data from the principal published studies describing the use of terpenes and their biological applications as antifungals.

Os terpenóides, também chamados terpenos ou isoprenóides, são uma família de produtos naturais encontrados em todos os organismos vivos. Muitas plantas são produtoras destes metabolitos secundários, que constituem uma grande parte dos óleos essenciais. Uma das características mais importantes é que os compostos são voláteis, têm odor e podem ser utilizados numa variedade de aplicações em diferentes segmentos industriais ou na medicina tradicional. O Brasil tem uma flora rica e diversificada que pode ser utilizada como fonte de pesquisa para a obtenção de novas moléculas. Dentro desta flora, vale a pena mencionar a Caatinga como um bioma exclusivamente brasileiro que possui plantas adaptadas a uma série de condições climáticas e, portanto, um armazém de compostos a serem descritos. As infecções fúngicas são doenças cada vez mais comuns, devido a isso existe uma grande procura de novos agentes com baixa toxicidade e efeitos secundários. Os cientistas devem procurar novas moléculas que exibam atividade antifúngica para o desenvolvimento de novos medicamentos contra as infecções fúngicas. Esta revisão visa analisar dados científicos dos principais estudos publicados que descrevem o uso de terpenóides e as suas aplicações biológicas como antifúngicos.

Identification of the tributary branches of the hepatic portal vein in the common sloth, Bradypus variegatus Schinz, 1825 (Pilosa: Bradypodidae) / [Identificação dos ramos tributários da veia porta hepática na preguiça-comum, Bradypus variegatus Schinz, 1825 (Pilosa: Bradypodidae)]

Bradypus variegatus, the common sloth, belongs to the Bradypodidae family, being considered a biological model to be applied in multidisciplinary research. This study was developed with the aim of being applied to clinical medicine and to the adequate management of the common sloth. Ten sloths were utilized, obtained post-natural death. The animals were fixed and to obtain the results, they were submitted to the dissection technique. For 80% of the animals, the portal vein originated from five tributaries, which were: the resulting vein from the anastomosis of the cardia vein, fundic vein, and the pyloric branches; the mesenteric trunk; the vein formed by the confluence of the stomach body branches and the cranial portion of the cavity of the cardia; the pyloric vein and splenic vein. While in 20% of the animals, the portal vein was comprised of six tributaries, because the fundic vein and cardia vein form two direct anastomoses, arriving at the portal vein two tributary vessels. This pattern differs in number and arrangement of branches when compared to the main domestic species. Therefore, the hepatic portal system is responsible for the drainage of the stomach, spleen, pancreas and intestines.

Bradypus variegatus, a preguiça-comum, pertence à família Bradypodidae, sendo considerada um modelo biológico a ser aplicado em pesquisas multidisciplinares. Este estudo foi desenvolvido a fim de ser aplicado à clínica médica e ao manejo adequado da preguiça-comum. Foram utilizadas 10 preguiças, obtidas após morte natural. Os animais foram fixados e, para a obtenção dos resultados, submeteram-se à técnica de dissecação. Em 80% das observações, a veia porta originou-se a partir de cinco tributárias, são elas: a veia resultante da anastomose da veia cárdia, da veia fúndica e dos ramos pilóricos; o tronco mesentérico; a veia formada a partir da confluência de ramos do corpo estomacal e da porção cranial da cavidade cárdica; a veia pilórica e a veia esplênica. Enquanto em 20% dos animais a veia porta é constituída por seis tributárias, a veia fúndica e a veia cárdica formam duas anastomoses diretas, chegando à veia porta dois vasos tributários. Esse padrão difere em número e em disposição dos ramos, quando comparado ao das principais espécies domésticas. Portanto, o sistema porta hepático é responsável pela drenagem do estômago, do baço, do pâncreas e dos intestinos.

In vivo phagocytosis and hematology in Astyanax altiparanae, a potential model for surrogate technology

Although the potential of surrogate propagation technology for aquaculture and conservation of Neotropical fish, the poor understanding of the host immune system may results in rejection and destruction of the donor material. Thus, it is necessary to study and to develop methods to evaluate the effects of immunosuppressive drugs employment and to evaluate the immunocompatibility between donor and receptor. Thus, the present study aimed to optimize a methodology to assess in vivo phagocytosis in Astyanax altiparanae using Saccharomyces cerevisiae and to evaluate their hematological response resultant from the inflammatory induction. To this, S. cerevisiae were labeled with Congo red and injected in the coelomic cavity of A. altiparanae at the concentration of 2.5 x 106 cells mL-1. A PBS solution and a non-injected group were kept as control. Fish blood was sampled and the phagocytic capacity and index were determined at 1, 2, 3 and 6 h post-injection (hpi). The yeast injection successfully stimulated phagocytosis, with the best result for phagocytosis assessment after 2 hpi. Moreover, it was achieved a high traceability of phagocytized and non-phagocytized yeast under optic microscopy analysis due to the Congo red labeling. The hematological profile was similar to usually observed in early infections, indicating lymphocyte migration to inflammatory site and increase in number of circulating phagocytes due to natural response to inflammatory stimulus. In conclusion, our method was efficient to assess in vivo phagocytosis in A. altiparanae and will be an important tool to evaluate the efficacy of immunosuppressive drugs in this species. Additionally, these results may serve as support for further studies in fish immunocompetence, both in laboratory and in field conditions.(AU)

Apesar do potencial apresentado pela tecnologia de propagação mediada para a aquicultura e conservação de peixes Neotropicais, o pobre entendimento do sistema imune do hospedeiro pode resultar na rejeição e destruição do material do doador. Com isso, se fazem necessários o estudo e o desenvolvimento de métodos para análise tanto dos efeitos de drogas imunossupressoras quanto para a avaliação da imunocompatibilidade entre doadores e receptores. Logo, o presente estudo teve como objetivo aperfeiçoar um método para analisar a fagocitose in vivo em Astyanax altiparanae usando Saccharomyces cerevisiae marcado e avaliar seu perfil hematológico resultante da indução inflamatória. Para isso, S. cerevisiae foram marcados com vermelho Congo e injetados na cavidade celomática dos A. altiparanae na concentração de 2,5 x 106 células.mL-1. Peixes injetados com PBS e peixes não injetados foram mantidos como controle. Sangue foi colhido e a capacidade fagocítica e o índice fagocítico foram determinados após 1, 2, 3 e 6 horas após à injeção (hpi). A injeção de levedura estimulou a fagocitose com sucesso, com o melhor resultado atingido após 2 hpi. Ainda, foi observada uma alta rastreabilidade das leveduras fagocitadas e não fagocitadas sob microscopia óptica devido à marcação com vermelho Congo. O perfil hematológico foi similar ao observado usualmente em infecções recém-induzidas, indicando migração de linfócitos ao sítio inflamatório e aumento no número de fagócitos circulantes devido à resposta natural ao estímulo inflamatório. Como conclusão, nosso método foi eficiente para analisar a fagocitose in vivo em A. altiparanae e será uma ferramenta importante para a avaliação de eficácia de drogas imunossopressoras para esta espécie. Em adição, estes resultados podem contribuir para futuros estudos em imunocompetência em peixes, tanto em âmbito laboratorial quanto a campo.(AU)

In vivo phagocytosis and hematology in Astyanax altiparanae, a potential model for surrogate technology

Abstract Although the potential of surrogate propagation technology for aquaculture and conservation of Neotropical fish, the poor understanding of the host immune system may results in rejection and destruction of the donor material. Thus, it is necessary to study and to develop methods to evaluate the effects of immunosuppressive drugs employment and to evaluate the immunocompatibility between donor and receptor. Thus, the present study aimed to optimize a methodology to assess in vivo phagocytosis in Astyanax altiparanae using Saccharomyces cerevisiae and to evaluate their hematological response resultant from the inflammatory induction. To this, S. cerevisiae were labeled with Congo red and injected in the coelomic cavity of A. altiparanae at the concentration of 2.5 x 106 cells mL-1. A PBS solution and a non-injected group were kept as control. Fish blood was sampled and the phagocytic capacity and index were determined at 1, 2, 3 and 6 h post-injection (hpi). The yeast injection successfully stimulated phagocytosis, with the best result for phagocytosis assessment after 2 hpi. Moreover, it was achieved a high traceability of phagocytized and non-phagocytized yeast under optic microscopy analysis due to the Congo red labeling. The hematological profile was similar to usually observed in early infections, indicating lymphocyte migration to inflammatory site and increase in number of circulating phagocytes due to natural response to inflammatory stimulus. In conclusion, our method was efficient to assess in vivo phagocytosis in A. altiparanae and will be an important tool to evaluate the efficacy of immunosuppressive drugs in this species. Additionally, these results may serve as support for further studies in fish immunocompetence, both in laboratory and in field conditions.

Resumo Apesar do potencial apresentado pela tecnologia de propagação mediada para a aquicultura e conservação de peixes Neotropicais, o pobre entendimento do sistema imune do hospedeiro pode resultar na rejeição e destruição do material do doador. Com isso, se fazem necessários o estudo e o desenvolvimento de métodos para análise tanto dos efeitos de drogas imunossupressoras quanto para a avaliação da imunocompatibilidade entre doadores e receptores. Logo, o presente estudo teve como objetivo aperfeiçoar um método para analisar a fagocitose in vivo em Astyanax altiparanae usando Saccharomyces cerevisiae marcado e avaliar seu perfil hematológico resultante da indução inflamatória. Para isso, S. cerevisiae foram marcados com vermelho Congo e injetados na cavidade celomática dos A. altiparanae na concentração de 2,5 x 106 células.mL-1. Peixes injetados com PBS e peixes não injetados foram mantidos como controle. Sangue foi colhido e a capacidade fagocítica e o índice fagocítico foram determinados após 1, 2, 3 e 6 horas após à injeção (hpi). A injeção de levedura estimulou a fagocitose com sucesso, com o melhor resultado atingido após 2 hpi. Ainda, foi observada uma alta rastreabilidade das leveduras fagocitadas e não fagocitadas sob microscopia óptica devido à marcação com vermelho Congo. O perfil hematológico foi similar ao observado usualmente em infecções recém-induzidas, indicando migração de linfócitos ao sítio inflamatório e aumento no número de fagócitos circulantes devido à resposta natural ao estímulo inflamatório. Como conclusão, nosso método foi eficiente para analisar a fagocitose in vivo em A. altiparanae e será uma ferramenta importante para a avaliação de eficácia de drogas imunossopressoras para esta espécie. Em adição, estes resultados podem contribuir para futuros estudos em imunocompetência em peixes, tanto em âmbito laboratorial quanto a campo.

In vivo phagocytosis and hematology in Astyanax altiparanae, a potential model for surrogate technology

Abstract Although the potential of surrogate propagation technology for aquaculture and conservation of Neotropical fish, the poor understanding of the host immune system may results in rejection and destruction of the donor material. Thus, it is necessary to study and to develop methods to evaluate the effects of immunosuppressive drugs employment and to evaluate the immunocompatibility between donor and receptor. Thus, the present study aimed to optimize a methodology to assess in vivo phagocytosis in Astyanax altiparanae using Saccharomyces cerevisiae and to evaluate their hematological response resultant from the inflammatory induction. To this, S. cerevisiae were labeled with Congo red and injected in the coelomic cavity of A. altiparanae at the concentration of 2.5 x 106 cells mL-1. A PBS solution and a non-injected group were kept as control. Fish blood was sampled and the phagocytic capacity and index were determined at 1, 2, 3 and 6 h post-injection (hpi). The yeast injection successfully stimulated phagocytosis, with the best result for phagocytosis assessment after 2 hpi. Moreover, it was achieved a high traceability of phagocytized and non-phagocytized yeast under optic microscopy analysis due to the Congo red labeling. The hematological profile was similar to usually observed in early infections, indicating lymphocyte migration to inflammatory site and increase in number of circulating phagocytes due to natural response to inflammatory stimulus. In conclusion, our method was efficient to assess in vivo phagocytosis in A. altiparanae and will be an important tool to evaluate the efficacy of immunosuppressive drugs in this species. Additionally, these results may serve as support for further studies in fish immunocompetence, both in laboratory and in field conditions.

Resumo Apesar do potencial apresentado pela tecnologia de propagação mediada para a aquicultura e conservação de peixes Neotropicais, o pobre entendimento do sistema imune do hospedeiro pode resultar na rejeição e destruição do material do doador. Com isso, se fazem necessários o estudo e o desenvolvimento de métodos para análise tanto dos efeitos de drogas imunossupressoras quanto para a avaliação da imunocompatibilidade entre doadores e receptores. Logo, o presente estudo teve como objetivo aperfeiçoar um método para analisar a fagocitose in vivo em Astyanax altiparanae usando Saccharomyces cerevisiae marcado e avaliar seu perfil hematológico resultante da indução inflamatória. Para isso, S. cerevisiae foram marcados com vermelho Congo e injetados na cavidade celomática dos A. altiparanae na concentração de 2,5 x 106 células.mL-1. Peixes injetados com PBS e peixes não injetados foram mantidos como controle. Sangue foi colhido e a capacidade fagocítica e o índice fagocítico foram determinados após 1, 2, 3 e 6 horas após à injeção (hpi). A injeção de levedura estimulou a fagocitose com sucesso, com o melhor resultado atingido após 2 hpi. Ainda, foi observada uma alta rastreabilidade das leveduras fagocitadas e não fagocitadas sob microscopia óptica devido à marcação com vermelho Congo. O perfil hematológico foi similar ao observado usualmente em infecções recém-induzidas, indicando migração de linfócitos ao sítio inflamatório e aumento no número de fagócitos circulantes devido à resposta natural ao estímulo inflamatório. Como conclusão, nosso método foi eficiente para analisar a fagocitose in vivo em A. altiparanae e será uma ferramenta importante para a avaliação de eficácia de drogas imunossopressoras para esta espécie. Em adição, estes resultados podem contribuir para futuros estudos em imunocompetência em peixes, tanto em âmbito laboratorial quanto a campo.

Produção de embriões em ovinos Morada Nova e Somalis Brasileira / Embryo production in Morada Nova and Somalis Brasileira ewes

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

Produção de embriões em ovinos Morada Nova e Somalis Brasileira / Embryo production in Morada Nova and Somalis Brasileira ewes

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

Uroguanylin induces electroencephalographic spikes in rats / Uroguanilina induz potenciais em espiga no eletroencefalograma de ratos

Uroguanylin (UGN) is an endogenous peptide that acts on membrane-bound guanylate cyclase receptors of intestinal and renal cells increasing cGMP production and regulating electrolyte and water epithelial transport. Recent research works demonstrate the expression of this peptide and its receptor in the central nervous system. The current work was undertaken in order to evaluate modifications of electroencephalographic spectra (EEG) in anesthetized Wistar rats, submitted to intracisternal infusion of uroguanylin (0.0125 nmoles/min or 0.04 nmoles/min). The current observations demonstrate that 0.0125 nmoles/min and 0.04 nmoles/min intracisternal infusion of UGN significantly enhances amplitude and frequency of sharp waves and evoked spikes (p = 0.03). No statistical significance was observed on absolute alpha and theta spectra amplitude. The present data suggest that UGN acts on bioelectrogenesis of cortical cells by inducing hypersynchronic firing of neurons. This effect is blocked by nedocromil, suggesting that UGN acts by increasing the activity of chloride channels.(AU)

A uroguanilina (UGN) é um peptídeo endógeno que age em receptores do tipo guanilato ciclase de membrana de células intestinais e renais aumentando a produção de GMPc e regulando o transporte epitelial de eletrólitos e água. Pesquisas recentes demonstraram a expressão deste peptídeo e de seus receptores no sistema nervosa central. O presente trabalho foi realizado com objetivo de avaliar possíveis mudanças no espectro do eletroencefalograma (EEG) de ratos Wistar anestesiados, submetidos à infusão intracisternal de uroguanilina (0.0125 nmoles/min or 0.04 nmoles/min). Os resultados apresentados no corrente trabalho demonstram que a infusão intracisternal de ambas as doses de UGN aumenta significativamente a amplitude e frequência das espículas (p = 0.03). Não foram encontradas diferenças estatísticas na amplitude absoluta dos espectros alfa ou teta. Os dados apresentados neste trabalho mostram que a UGN age na bioeletrogênese de células corticais induzindo disparo hipersincrônico de neurônios. Este efeito é bloqueado por nedocromil, sugerindo que UGN atua pelo aumento de atividade de canais de cloreto.(AU)

Study of irradiated bothropstoxin-1 with60Co gamma rays: immune system behavior

Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules. Very promising results were obtained when crude animal venoms, as well as isolated toxins, were treated with 60Co gamma rays, yielding toxoids with good immunogenicity. The achievement of modified antigens with lower toxicity and preserved or improved immunogenicity can be very useful. Ionizing radiation has already been proven to be a powerful tool to attenuate snake venom toxicity without affecting, and even increasing, their immunogenic properties. However, little is known about the modifications that irradiated molecules undergo and even less about the immunological response that such antigens elicit. In the present work, we investigated the immunological behavior of bothropstoxin-1, a K49 phospholipase, before and after irradiation. Structural modifications of the toxin were analyzed by SDS-PAGE. Isogenic mice were immunized with either the native or the irradiated toxin. The circulating antibodies were isotyped and titrated by ELISA. According to our data, irradiation promoted structural modifications in the toxin characterized by higher molecular weight forms of proteins (aggregates and oligomers). The results also indicated that irradiated toxins were immunogenic and antibodies elicited by them were able to recognize the native toxin in ELISA. These findings suggest that irradiation of toxic proteins can promote significant modifications in their structures; however they still retain many of the original antigenic and immunological properties of native proteins. Also, our data indicate that irradiated proteins induce higher titers of IgG2a and IgG2b, suggesting that Th1 cells are predominantly involved in the immune response.

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 µg/g in the liver and 51.2 µg/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 µg/g in the liver and 51.2 µg/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.(AU)

Laboratory evaluation of young ovines inoculated with natural or 60co-irradiated Crotalus durissus terrificus venom during hyperimmunization process

Laboratory profile of young ovines was studied in order to evaluate and compare their antiserum production from natural and Cobalt-60 irradiated Crotalus durissus terrificus (C.d.t.) venoms. The parameters analyzed included complete blood count, and urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin serum measurements. Three groups of six animals each were used. Group 1 (G1) received natural C.d.t. venom; Group 2 (G2) received irradiated C.d.t. venom; and Group 3 (G3) was used as control and did not receive venom, only adjuvants, using seven venom inoculations. During the experimental period, animals were fortnightly weighed. According to clinical and weight evaluation, sheep in post-weaning phase showed no changes in their physiological profiles but had excellent weight gain. The parameters analyzed were not statistically different (p 5%) among the groups tested. The hyperimmunization process was successfully accomplished with the production of specific antibodies against Crotalus durissus terrificus venom. Results bring a new possibility of utilizing ovines in the commercial production of anticrotalic serum, which may be used to treat human and animal envenomation. Its production cost may be reduced by subsequent use of hyperimmunized sheep for human consumption.

Immunological assessment of mice hyperimmunized with native and Cobalt-60-irradiated Bothrops venoms

ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated (60Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom.

Immunization with native and cobalt 60-irradiated Crotalus durissus terrificus venom in swiss mice: assessment of the neutralizing potency of antisera

ELISA was used to evaluate, follow, and compare the humoral immune response of Swiss mice during hyperimmunization with natural and Cobalt 60-irradiated (60Co) Crotalus durissus terrificus venom. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by "in vivo" challenge and the side effects were assessed. The animals immunization with one LD50 of the venom was on days one, 15, 21, 30, and 45, when blood samples were collected; the challenges occurred on the 60th day. Results showed that ELISA was efficient in evaluating, following, and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co irradiation. Serum produced from Crotalus durissus terrificus irradiated venom showed higher potency and neutralization capacity than that from natural venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom.

Avaliação toxicológica do extrato acetato de etila de Spigelia anthelmia Linn, em ratos e camundongos

O extrato acetato de etila de Spigelia anthelmia (EASa) mostrou formalmente ser altamente eficaz contra o desenvolvimentolarvar e a eclosão de ovos de Haemonchus contortus, um importante parasito de ruminantes, in vitro. A DL1 e a DL10 de EASaforam administradas subcrônica e cronicamente pela via oral em ratos wistar e o perfil bioquímico foi comparado antes e apóscada tratamento e com veículo. Vários órgãos foram coletados e processados para análise histopatológica. Os parâmetroshematológicos foram avaliados antes e depois da administração de EASa durante 30 dias. E os efeitos do EASa administradopela via oral durante o período embriogênico ou organogênico a camundongas gestantes foram estudados. Os efeitos diretosde EASa, in vivo, foram calculados na pressão sangüínea arterial média e no eletrocardiograma (ECG), e in vitro no coraçãoisolado e no átrio isolado de ratos. A administração de EASa não afetou qualquer parâmetro bioquímico, hematológico oureprodutivo estudado. EASa induziu um efeito hipotensivo de curto prazo em ratos normotensivos sem qualquer alteraçãoconcomitante nos parâmetros do ECG. As maiores doses de EASa induziram uma significante diminuição da amplitude decontração do coração e átrio direito. EASa é desprovido de toxicidade significante e tem leves efeitos no sistema cardiovascular.

Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

Natural (NV) and Cobalto60-irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 mM, pH 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count (CBC), total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/ml indicates that slight leukocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the NV group, showing its greater toxicity.

Avaliação toxicológica do extrato acetato de etila de Spigelia anthelmia Linn, em ratos e camundongos

O extrato acetato de etila de Spigelia anthelmia (EASa) mostrou formalmente ser altamente eficaz contra o desenvolvimentolarvar e a eclosão de ovos de Haemonchus contortus, um importante parasito de ruminantes, in vitro. A DL1 e a DL10 de EASaforam administradas subcrônica e cronicamente pela via oral em ratos wistar e o perfil bioquímico foi comparado antes e apóscada tratamento e com veículo. Vários órgãos foram coletados e processados para análise histopatológica. Os parâmetroshematológicos foram avaliados antes e depois da administração de EASa durante 30 dias. E os efeitos do EASa administradopela via oral durante o período embriogênico ou organogênico a camundongas gestantes foram estudados. Os efeitos diretosde EASa, in vivo, foram calculados na pressão sangüínea arterial média e no eletrocardiograma (ECG), e in vitro no coraçãoisolado e no átrio isolado de ratos. A administração de EASa não afetou qualquer parâmetro bioquímico, hematológico oureprodutivo estudado. EASa induziu um efeito hipotensivo de curto prazo em ratos normotensivos sem qualquer alteraçãoconcomitante nos parâmetros do ECG. As maiores doses de EASa induziram uma significante diminuição da amplitude decontração do coração e átrio direito. EASa é desprovido de toxicidade significante e tem leves efeitos no sistema cardiovascular.

Avaliação toxicológica do extrato acetato de etila de Spigelia anthelmia Linn, em ratos e camundongos

O extrato acetato de etila de Spigelia anthelmia (EASa) mostrou formalmente ser altamente eficaz contra o desenvolvimentolarvar e a eclosão de ovos de Haemonchus contortus, um importante parasito de ruminantes, in vitro. A DL1 e a DL10 de EASaforam administradas subcrônica e cronicamente pela via oral em ratos wistar e o perfil bioquímico foi comparado antes e apóscada tratamento e com veículo. Vários órgãos foram coletados e processados para análise histopatológica. Os parâmetroshematológicos foram avaliados antes e depois da administração de EASa durante 30 dias. E os efeitos do EASa administradopela via oral durante o período embriogênico ou organogênico a camundongas gestantes foram estudados. Os efeitos diretosde EASa, in vivo, foram calculados na pressão sangüínea arterial média e no eletrocardiograma (ECG), e in vitro no coraçãoisolado e no átrio isolado de ratos. A administração de EASa não afetou qualquer parâmetro bioquímico, hematológico oureprodutivo estudado. EASa induziu um efeito hipotensivo de curto prazo em ratos normotensivos sem qualquer alteraçãoconcomitante nos parâmetros do ECG. As maiores doses de EASa induziram uma significante diminuição da amplitude decontração do coração e átrio direito. EASa é desprovido de toxicidade significante e tem leves efeitos no sistema cardiovascular.