RESUMO Objetivou-se avaliar a condição metabólica e estrutural das células espermáticas bovinas após congelação, com adição prévia de IGF-I e insulina no meio diluidor seminal. Os ejaculados de seis touros Nelore foram submetidos a quatro tratamentos: controle; insulina (100µUI/mL); IGF-I (150ng/mL) e insulina + IGF-I (50µUI/mL e 75ng/mL, respectivamente). Após a congelação, realizaram-se os testes de termorresistência rápida, coloração pelo corante azul de tripan e Giemsa, além da análise computadorizada da motilidade espermática, da integridade das membranas plasmática e acrossomal, e da peça intermediária por meio de sondas fluorescentes. O teste de termorresistência rápida apresentou efeito dentro do tempo de cada tratamento, mas não entre os tratamentos. Na análise computadorizada da motilidade espermática, foram observados movimento, motilidade e velocidade espermáticos; não houve efeitos dos tratamentos sobre qualquer uma dessas variáveis. Respostas iguais foram obtidas com as sondas fluorescentes e o corante azul de tripan/Giemsa. A adição de insulina e IGF-I, de forma isolada ou combinada, ao meio diluidor para congelação de sêmen não produziu efeitos sobre as condições metabólica e estrutural das células espermáticas.

ABSTRACT This study aimed to evaluate the metabolic and structural condition of the spermatic bovine cells after the freezing, with addition, previously, of IGF-I and Insulin in the seminal thinner medium. The semen of 6 Nellore bulls were submitted to four treatments: Control, Insulin (100µUI/mL); IGF-I (150ng/mL) and Insulin + IGF-I (50µUI/mL and 75ng/mL, respectively). After freezing, rapid resistance tests, Tripan and Giemsa Blue staining, and computerized analysis of sperm motility and integrity of the plasma and acrosomal membranes and the intermediate part were performed by fluorescent probes. The term rapid resistance test had effect within the time of each treatment, but not between treatments. In the computer analysis of sperm motility, sperm movement, motility and velocity no effects of treatments were observed on any of these variables. The same results were obtained with the fluorescent probes and the Blue dye Trypan / Giemsa. The addition of Insulin and IGF-I, alone or in combination, to the semen freezing dilution medium had no effect on the metabolic and structural condition of sperm cells.

RESUMO Este estudo tem como objetivo avaliar a adição da vitamina C, da glutationa reduzida e do trolox sobre características espermáticas do sêmen refrigerado de suínos. Seis cachaços foram coletados pela técnica de mão enluvada (10 coletas/animal). O sêmen foi diluído em MR-A®. Após as avaliações prévias, os tratamentos foram adicionados: grupo controle: apenas diluidor; grupo vitamina C: 200μM/mL de vitamina C; grupo trolox: 200μM/mL de trolox; grupo glutationa: 2.5mM/mL de glutationa reduzida. O sêmen foi armazenado em caixas térmicas e alocado dentro do refrigerador a 15oC e avaliado nos tempos zero, 12, 48 e 72 horas . Após 30 horas de incubação, cada tratamento foi dividido em duas frações iguais e adicionou-se a mesma concentração de antioxidantes em uma das partes. Os resultados demonstram que a suplementação de glutationa reduzida preserva a motilidade espermática após 24 horas, bem como tem maior percentual de acrossoma intacto na presença desse antioxidante. Não houve efeito ao se adicionar uma segunda dose dos antioxidantes. Em conclusão, o acréscimo da glutationa reduzida ao diluidor de sêmen suíno é uma alternativa promissora para melhor preservação das características espermáticas, e a adição da segunda dose dos antioxidantes durante o armazenamento é prejudicial ao sêmen.

ABSTRACT This study aimed to evaluate the addition of Vitamin C, reduced Glutathione and trolox on sperm characteristics of pork refrigerated semen. Six pigs were collected through the technique of gloved hand (10 ejaculates/animals). The semen was diluted in MR-A®. After the previous evaluations, the treatments were added: Control group: diluent only; Vitamin C Group: 200μM/mL Vitamin C; Trolox Group: 200μM/mL Trolox; Glutathione group: 2.5mM/ml Reduced glutathione. The semen was stored in thermal boxes and placed inside the refrigerator at 15oC and evaluated at D0, 12, 48, 72 hours. After 30 hours of incubation, each treatment was divided into two equal fractions and the same concentration of antioxidants was added in one of the parts. The results show that reduced glutathione supplementation preserves sperm motility after 24 hours but also has a higher percentage of acrosome intact in the presence of this antioxidant. There was no effect of adding a second dose of the antioxidants. In conclusion, the addition of reduced Glutathione to the swine semen diluent is a promising alternative for better preservation of sperm characteristics and the addition of the second dose of antioxidants during storage is detrimental to semen.

This study aimed to evaluate the chemical composition, fractionation of nitrogen compounds and carbohydrates, digestibility parameters, gas production kinetics and ruminal fermentation of fruit seed cakes. The experiment was conducted in a completely randomized design with three fruit seed cakes (treatments) and four replications. The cakes were made of seeds of custard apple (Annona squamosa), soursop (Annona muricata) and passion fruit (Passiflora edulis), which were collected in a biodiesel plant. The passion fruit seed cake presented the highest content of dry matter, neutral detergent fibre crude protein (NDFcp), acid detergent fibre (ADF), lignin and total carbohydrates (P<0.05). The custard apple seed cake presented high proportions of crude protein, condensed tannins and digestible energy (P<0.05). The soursop seed cake presented higher values for ether extract and total digestible nutrients (P<0.05). For potential degradability, the authors detected a difference between the custard apple, soursop and passion fruit seed cakes. The total volume of gases was higher for custard apple and soursop seed cakes. Custard apple seed cake presented higher proportions of in vitro degradation parameters (P<0.05). Thus this cake could provide greater nutrient levels when supplied in ruminant diets, followed by soursop seed cake and passion fruit seed cake.

RESUMO O número de transplantes de órgãos e tecidos em humanos e animais tem crescido significativamente nos últimos anos, principalmente após o advento de técnicas modernas e mais seguras indutoras de imunossupressão. Objetiva-se com o presente estudo avaliar macro e microscopicamente o alotransplante parcial de bexiga a fresco em coelhos, utilizando como agente imunomodulador células-tronco mesenquimais derivadas do tecido adiposo (ADSC) alogênicas. Foram utilizados 25 coelhos, sendo um deles macho e doador das ADSCs, e os outros 24 eram fêmeas, submetidas a alotransplante parcial de bexiga, tratadas com ciclosporina (GCi) ou células-tronco mesenquimais (GCe). Conclui-se que o GCe teve melhor aceitação histológica do implante em relação ao GCi aos 30 dias de avaliação.

ABSTRACT The number of organ and tissue transplantation in humans and animals has grown significantly recently, especially after the advent of modern and safer techniques of immunosuppression. The objective of this study was to evaluate macro and microscopically partial urinary bladder fresh allograft in rabbits, using as immunomodulatory agent cyclosporine or allogenic adipose tissue derived mesenchymal stem cells (ADSCs). For this purpose, 25 rabbits were used. One male was the donor of ADSCs; 24 females received a partial urinary bladder allograft and were treated with cyclosporine (GCi) or mesenchymal stem cells (GCe). We conclude that the GCe group had better histological acceptance of the implant than GCi group at 30 days evaluation.

Torch ginger Etlingera elatior (Jack) R.M. Smith is a tropical herb-like, rhizome-like and perennial plant that possesses inflorescences that are both pretty and showy at different colour shades, and have a great visual appeal. Producing seedlings at a high enough level when done conventionally by the division of tussocks, has high costs and phytosanitary problems. The objective of this study was to evaluate the induction of somatic embryogenesis and its potential for subsequent plant regeneration in torch ginger. Rhizomes, leaves and roots from the seedlings grown in vitro were used as explant segments. The different explants were cultured on MS medium supplemented with 30 g L-1 sucrose, and it was solidified with 5.5 g L-1 of agar. The effect of 2.4-D and picloram concentrations of 0.0, 1.0, 2.0 and 4.0 mg L-1 was studied. Astructural analysis was performed by scanning electron microscopy (SEM). Cytochemical analysis were performed to detect nucleeous with carmine acetic, starch with lugol and lipids with Sudan III, and a flow cytometry analysis to verify the stability of ploidy in the callus. Results indicate that the culture medium with 2.4-D yielded the induction of embryogenic callus with the characteristics of the rhizome segments of the explant. Regarding the ultrastructural analysis of embryogenic callus characteristics, the cells showed a similar isodiametric form as somatic embryos in the globular stage and cytochemical analysis. The presence of callus cells in pro-embryogenic explants containing each of the regulators was confirmed. In the cytometry analysis, the average DNA content of callus ranged from 5.43 to 10.15 pg, and the coefficients of variation varied between 0.3 to 2.26. Callus induced with 2.4-D showed 86 % stability, and with picloram was 64 %. The callus formation was more evident after application of 2.4 D. This is the first study of embryogenic potential in cells of the torch ginger, and showed that 2,4-D induced callus with higher starch content and genetically stable.

El jengibre antorcha Etlingera elatior (Jack) R. M. Smith es una planta tropical perenne, similar al rizoma, que tiene inflorescencias que son bellas y llamativas, de diferentes colores y matices y de un gran atractivo visual. La producción de plántulas en un nivel lo suficientemente alto cuando se hace convencionalmente utilizando la división de matas tiene altos costos y problemas fitosanitarios. El objetivo de este estudio fue evaluar la inducción de la embriogénesis somática y su uso potencial para la reproducción posterior de plantas de jengibre antorcha. Los rizomas, hojas y raíces de las plántulas cultivadas in vitro se utilizaron como segmentos de explantes. Los diferentes explantes se cultivaron en un medio MS al que se le agregaron 30 g L-1 de sacarosa y fue solidificado con 5.5 g de agar L-1. Se estudió el efecto del 2.4-D y picloram en concentraciones de 0.0, 1.0, 2.0 y 4.0 mg L-1. El análisis estructural se llevó a cabo por microscopía electrónica de barrido (SEM). Se realizaron análisis citoquímicos para detectar núcleos con carmín acético, almidón con lugol y lípidos con Sudán III; y un análisis de citometría de flujo para verificar la estabilidad de la ploidía en el callo. Los resultados indicaron que el medio de cultivo con 2.4-D produjo la inducción de callo embriogénico con las características de los segmentos de rizomas del explante. En el análisis ultraestructural de las características del callo embriogénico, las células mostraron una forma isodiamétrica similar a embriones somáticos en etapa globular y análisis citoquímico. Se confirmó la presencia de células de callo en explantes pro- embriogénicos conteniendo cada uno de los reguladores. En el análisis de citometría, el contenido medio de ADN del callo varió de 5.43 a 10.15 pg, y los coeficientes de variación oscilaron entre 0.3 y 2.26. El 2.4-D indujo callos con estabilidad de 86 %, y con picloram fue 64 %. La formación de callo fue más evidente después de aplicar 2.4-D. Éste es el primer estudio del potencial embriogénico de células del jengibre antorcha, y mostró que el 2.4-D indujo la formación de callo con mayor contenido de almidón y genéticamente estable.

Entre os meses de março de 2000 e dezembro de 2005, 87 espécimes de L. gastrophysus foram obtidos de supermercados e empresas de exportação localizadas nos municipios de Cabo Frio, Niteroi, Duque de Caxias e Rio de Janeiro. Os peixes foram necropsiados, filetados e seus órgãos investigados para presença de helmintos. Dois espécimes (2,3%) estavam parasitados por plerocecoides de Diphyllobothrium Cobbold, 1858 na cavidade abdominal e serosa do intestino, com intensidade média de um parasito por peixe e abundância média de 0,02, mostrando baixo potencial infectivo. Este é o primeiro registro de plerocercoides de Diphyllobothrium sp. no peixe sapo-pescador.

A espécie vegetal Alpinia zerumbet (Pers.) B.L.Burtt & R.M. Sm. é popularmente empregada para o tratamento de diversas enfermidades, entre elas a hipertensão. Avaliar a composição química, a atividade antihipertensiva e ação na hipertrofia cardíaca do óleo essencial das folhas de Alpinia zerumbet (OEAZ) em ratos foram os objetivos deste estudo. O OEAZ, obtido por hidrodestilação em aparelho Clevenger, teve sua composição química analisada em cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Foram identificados 14 constituintes, sendo terpinen-4-ol (37,45%) o majoritário, seguido pelos óxido de cariofileno (7,56%), trans-hidrato de sabineno (6,61%) e 1,8-cineol (4,02%). A avaliação cardiovascular foi feita após o tratamento crônico de ratos espontaneamente hipertensos (SHR) e seus respectivos controles, ratos Wistar-Kyoto (WKY). Os dados hemodinâmicos revelaram redução da pressão arterial média (PAM) no grupo tratado (SHRP: 160 ± 7 mm Hg; p<0,01) em relação ao não tratado (SHR: 180 ± 5 mm Hg). A relação entre peso do ventrículo esquerdo e peso corporal (VE/PC) do SHRP (2,50 ± 0,03 mg g-1; p<0,01) mostrou-se inferior ao SHR (2,61 ± 0,01 mg g-1), confirmando a redução da hipertrofia cardíaca (HC). Os dados de PAM e VE/PC dos animais SHRP foram estatisticamente diferentes quando comparados com os ratos controle (WKY: 116 ± 2 mm Hg e WKYP: 119 ± 4 mm Hg; p<0,05; WKY: 2,15 ± 0,04 mg g-1 e WKYP: 2,17 ± 0,04 mg g-1 ; p<0,01), indicando não ter havido normalização dos mesmos. Conclui-se que o tratamento crônico com OEAZ foi capaz de determinar redução, mas não a normalização, da PAM e da HC de ratos SHR, provavelmente pela presença dos componentes terpinen-4-ol e 1,8-cineol. Estudos com doses maiores ou período de tratamento superior são necessários para avaliar a possibilidade de o OEAZ normalizar os parâmetros analisados (PAM e HC).

Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. is traditionally employed to treat several diseases such as hypertension. The aim of this study was to evaluate the chemical composition, the anti-hypertensive activity and the capacity to reduce cardiac hypertrophy of the essential oil of A. zerumbet leaves (EOAZ) in rats. EOAZ was obtained through hydrodistillation in Clevenger apparatus and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Several constituents (14) were identified, terpen-4-ol (37.45%) being the major component, followed by caryophyllene oxide (7.56%), trans-sabinene hydrate (6.61%) and 1,8-cineol (4.02%). The cardiovascular effect was investigated after chronic treatment with spontaneously hypertensive rats (SHR) and their respective controls, Wistar-Kyoto rats (WKY). The treated group showed a lower mean arterial pressure (MAP) (SHRP: 160 ± 7 mm Hg; p<0.01) than the untreated group (SHR: 180 ± 5 mm Hg). The ratio of left ventricle-to-body weight (LV/BW) for SHRP was lower (2.504 ± 0.03 mg g-1; p<0.01) than that for SHR (2.162 ± 0.01 mg g-1), confirming the cardiac hypertrophy (CH) reduction. There were significant differences in MAP and CH between SHRP animals and control rats (WKY: 116 ± 2 mm Hg and WKYP: 119 ± 4 mm Hg; p<0.05. WKY: 2.152 ± 0.04 mg g-1 and WKYP: 2.168 ± 0.04 mg g-1; p<0.01), indicating that these values were not normalized. Those data showed that the chronic treatment with EOAZ reduces MAP and CH in SHR probably due to the presence of the compounds terpinen-4-ol and 1,8-cineol. Studies with higher doses or longer treatment periods are necessary to evaluate whether EOAZ can reduce the analyzed parameters (MAP and CH) to normal values.

Relata-se o primeiro caso de infecção de perus, Meleagris gallopavo, por Cheilospirura hamulosa no Brasil, com base no achado de três espécimes fêmeas de C. hamulosa, inseridos na submucosa da moela de um peru, retirado de um lote de 15 aves, provenientes de criadouros domésticos no estado de Minas Gerais. A ocorrência foi de 6,7%. A ave parasitada não apresentava sinais clínicos. As lesões microscópicas da moela eram severas, caracterizando-se por intensos processos inflamatórios crônicos e difusos, com infiltrados mistos de granulócitos, estendendo-se à mucosa e às camadas musculares. Nessa área, foram observados fragmentos dos parasitos e perfuração da mucosa, com destruição das camadas musculares.

Cytochrome P450 (CYP) is a superfamily of enzymes involved in the metabolism of endogenous compounds and xenobiotics. CYP2A6 catalyzes the oxidation of nicotine and the activation of carcinogens such as aflatoxin B1 and nitrosamines. CYP2E1 metabolizes ethanol and other low-molecular weight compounds and can also activate nitrosamines. The CYP2A6 and CYP2E1 genes are polymorphic, altering their catalytic activities and susceptibility to cancer and other diseases. A number of polymorphisms described are ethnic-dependent. In the present study, we determined the genotype and allele frequencies of the main CYP2A6 and CYP2E1 polymorphisms in a group of 289 volunteers recruited at the Central Laboratory of Hospital Universitário Pedro Ernesto. They had been residing in the city of Rio de Janeiro for at least 6 months and were divided into two groups according to skin color (white and non-white). The alleles were determined by allele specific PCR (CYP2A6) or by PCR-RFLP (CYP2E1). The frequencies of the CYP2A6*1B and CYP2A6*2 alleles were 0.29 and 0.02 for white individuals and 0.24 and 0.01 for non-white individuals, respectively. The CYP2A6*5 allele was not found in the population studied. Regarding the CYP2E1*5B allele, we found a frequency of 0.07 in white individuals, which was statistically different (P < 0.05) from that present in non-white individuals (0.03). CYP2E1*6 allele frequency was the same (0.08) in both groups. The frequencies of CYP2A6*1B, CYP2A6*2 and CYP2E1*6 alleles in Brazilians are similar to those found in Caucasians and African-Americans, but the frequency of the CYP2E1*5B allele is higher in Brazilians.

O trematódeo digenético Paratanaisia bragai é referido pela primeira vez parasitando a rolinha-caldo-de-feijão, Columbina talpacoti, proveniente de área suburbana do Rio de Janeiro, Brasil. Embora com baixa prevalência (10%), a intensidade de infecção foi alta, considerando que 116 exemplares do parasito foram obtidos de um dos rins. Não foram observadas lesões macroscópicas. A análise histopatológica demonstrou grande dilatação dos dutos coletores renais, com destruição e achatamento das células epiteliais de revestimento, sem reação inflamatória. Os achados patológicos foram comparados aos anteriormente relatados para P. bragai em outros hospedeiros, desde a proposição da espécie em 1934.

This is the first report of the digenetic trematode Paratanaisia bragai infestation in a ruddy ground dove Columbina talpacoti, captured in a suburban area of Rio de Janeiro, Brazil. Although with a low prevalence (10%), the intensity of infection was high, considering that 116 worms were recovered from one of the kidneys. Gross lesions were not observed and histopathological analysis showed very dilated renal collecting ducts with destruction and flattening of the lining epithelial cells, without inflammatory reaction. The pathological findings were compared to those previously reported for P. bragai in other hosts, since the proposal of the species in 1934.

Coelhos foram infectados experimentalmente per os com 10 larvas L4 de Eustrogylides ignotus (n= 10) e 50 L3 de Contracaecum multipapillatum (n= 50) coletados em traíras (Hoplias malabaricus) naturalmente parasitadas a fim de se avaliar a patogenicidade induzida por essas espécies de nematóides em mamíferos. Dois coelhos (20%) infectados com E. ignotus morreram antes do quarto dia pós-infecção (um após 51 horas e outro após 78 horas). Seis coelhos (60%) mostraram-se inapetentes até o quinto dia após a infecção experimental. Não foram observados sinais clínicos nos coelhos infectados com C. multipapillatum. À necropsia, oito (80%) mostraram-se positivos para essa espécie de nematóide. Os coelhos infectados com E. ignotus apresentaram congestão gástrica, com formação de hematoma na parede gástrica em 60% dos casos. O peritônio estava congestionado em 20% dos animais, e em 10% dos casos foi observada a presença de abscesso. Todos os animais infectados apresentaram hiperemia da mucosa gástrica com gastrite hemorrágica provocada pela infecção com E. ignotus. Nas infecções com C. multipapillatum, a hiperemia foi seguida por ruptura da mucosa nos locais de fixação do parasito, e na mucosa gástrica observou-se infiltrado leucocitário, com necrose multifocais alcançando a submucosa. Nas infecções com E. ignotus foram constatadas lesões perfurantes em diversos órgãos, principalmente na parede gástrica, pâncreas e fígado, sempre na presença de processo inflamatório misto, intensamente fibrótico, com hemorragia e necrose.

Rabbits were infected per os with 10 Eustrongylides ignotus L4 and with 50 Contracaecum multipapillatum L3 per rabbit, recovered from naturally infected freshwater fishes (Hoplias malabaricus) in order to evaluate the patogenicity of these two nematode species in mammalian host. Two rabbits (20%) infected with E. ignotus died before the fourth day post-inoculation (one after 51 and the other after 78 hours). Six rabbits (60%) were inappetent until the fifth day following experimental inoculation. No clinical signs in rabbits inoculated with C. multipapillatum were observed; nevertheless, eight (80%) animals were positive for this nematode species. Rabbits inoculated with E. ignotus, had gastric congestion with hematoma of the gastric wall in 60% of the cases. Peritoneum was congested in 20% of the animals with the presence of peritoneal abscess in 10% of the cases. All inoculated animals showed hyperemia of the gastric mucosa with hemorrhagic gastritis due to infections with E. ignotus. In C. multipapillatum inoculated animals, the hyperemia was followed by disruption of the epithelial mucosa in the sites of parasite attachment. In the gastric mucosa, miscellaneous leukocitary infiltrates, with multifocal necrosis reaching the submucosa in the infections with C. multipapillatum were observed under bright field microscopy. Perforating lesions in several organs, mainly in the gastric wall, pancreas and liver, always in the presence of a mixed inflammatory process, intensely fibrous, with hemorrhage and necrosis were observed in animals infected with E. ignotus.

Cytochrome P450 (CYP) 2A enzymes are involved in the metabolism of numerous drugs and hormones and activate different carcinogens. Human CYP2A6, mouse CYP2A5 and rat CYP2A3 are orthologous enzymes that present high similarity in their amino acid sequence and share substrate specificities. However, different from the human and mouse enzyme, CYP2A3 is not expressed in the rat liver. There are limited data about expression of CYP2A3 in extrahepatic tissues and its regulation by typical CYP inducers. Therefore, the objective of the present study was to analyze CYP2A3 mRNA expression in different rat tissues by RT-PCR, and to study the influence of 3-methylcholanthrene, pyrazole and ß-ionone treatment on its expression. Male Wistar rats were divided into four groups of 5 rats each, and were treated ip for 4 days with 3-methylcholanthrene (25 mg/kg body weight), pyrazole (150 mg/kg body weight), ß-ionone (1 g/kg body weight), or vehicle. Total RNA was extracted from tissues and CYP2A3 mRNA levels were analyzed by semiquantitative RT-PCR. CYP2A3 mRNA was constitutively expressed in the esophagus, lung and nasal epithelium, but not along the intestine, liver, or kidney. CYP2A3 mRNA levels were increased in the esophagus by treatment with 3-methylcholanthrene and pyrazole (17- and 7-fold, respectively), in lung by pyrazole and ß-ionone (3- and 4-fold, respectively, although not statistically significant), in the distal part of the intestine and kidney by 3-methylcholanthrene and pyrazole, and in the proximal part of the intestine by pyrazole. CYP2A3 mRNA was not induced in nasal epithelium, liver or in the middle part of the intestine. These data show that, in the rat, CYP2A3 is constitutively expressed in several extrahepatic tissues and its regulation occurs through a complex mechanism that is essentially tissue specific.

Tracheal organ cultures (TOC) were prepared and used for evaluating four Brazilian isolates of infectious bronchitis virus (IBV). IBV field isolates and vaccine strains were titrated in TOC and results compared to those from chicken embrionated eggs. Serum neutralization (SN) employing IBV strain-specific serum was performed for evaluating relationships between isolates. Titration results of tests performed in TOC or eggs were in mutual agreement and were considered for validating the adapted TOC methodology as alternative for virological studies in our laboratory. Sera specific to M41 (Massachusetts) or A5968 (Connecticut) did neutralize their respective IBV strains only. Field strains 208 and 29-78 specific sera did neutralize Massachusetts serotype strains M41 and H120, but PM2 serum did only M41. Strain PM4 specific serum did not neutralize any of the reference IBV analyzed, including M41, A5968 and H120 and may indicate that the isolate is serologically different from the Massachusetts serotype, currently adopted for vaccine strains in Brazil.