Achatina fulica (Gastropoda: Pulmonata): Occurrence, environmental aspects and presence of nematodes in Sergipe, Brazil / Achatina fulica (Gastropoda: Pulmonata): ocorrencia, aspectos ambientais e presença de nematódeos em Sergipe, Brasil

Abstract The objective of this study was to identify the occurrence of Achatina fulica in the Great Aracaju-SE Territory. This research was developed during the months of June 2016 to May 2017. The plots methodology was used for the monthly collections, following the collection time of 20 minutes per area. The mollusks were measured and soon afterwards the parasitological analysis of the mucus and faeces was performed. Molecular identification was performed to evaluate the possible presence of Angiostrongylus sp. We observed that the occurrence of A. fulica was more representative in urban areas, especially in vacant lots with the presence of litter and decomposing materials. In the specimens of A. fulica examined were male and female nematodes of the genus Rhabditis, in the research municipalities, except in Barra dos Coqueiros. No parasites of the genus Angiostrongylus were found in the mollusks examined. The influence of the presence of litter on the plots associated with the frequency of A. fulica was statistically significant. There was a correlation between the influence of rainfall and the frequency of A. fulica. A significant relationship between moisture and frequency and nematode positivity was identified. It was also observed that the larger the mollusk, the greater the chances of being infected by nematodes. Scientific knowledge about the biology of the A. fulica species associated with human intervention such as the maintenance of land and adequate sanitary measures may be important factors for a better control of this invasive mollusk.

Resumo O objetivo deste estudo foi verificar a ocorrência de A. fulica no território da Grande Aracaju, Sergipe, avaliando a infecção destes moluscos por nematódeos, a influência das condições climáticas e das características do ambiente. Esta pesquisa foi desenvolvida durante os meses de junho de 2016 a maio de 2017. Foi utilizada a metodologia de parcelas para as coletas mensais de 20 min por área, um total de 15 parcelas distribuídas em 5 municípios. Os moluscos foram mensurados e logo em seguida realizada a análise parasitológica do muco e fezes. Foi realizada a identificação molecular para avaliar a possível presença de Angiostrongylus sp. Observamos que a ocorrência de A. fulica foi mais representativa em áreas urbanas, sobretudo em terrenos baldios com a presença de lixo e materiais em decomposição. Nos exemplares de A. fulica examinados foram encontrados nematódeos do gênero Rhabditis, nos municípios da pesquisa, exceto em Barra dos Coqueiros. Não foram encontrados parasitos do gênero Angiostrongylus nos moluscos examinados. A influência da presença de lixo nas parcelas associado à frequência da A. fulica mostrou-se estatisticamente significativa. Houve correlação entre a influência da chuva e a frequência de A. fulica. Foi identificada uma relação significativa entre a umidade e a frequência e a positividade para nematódeos. Foi também observado que quanto maior o molusco, maiores são as chances deste apresentar nematódeos. O conhecimento científico a respeito da biologia da espécie A. fulica associado à intervenção humana como, a manutenção de terrenos e medidas sanitárias adequadas podem ser fatores importantes para um controle malacológico mais adequado para esta espécie invasora.

Hematological and biochemical profiles of Mangalarga Marchador mares in the transition period bred on pasture / Perfis hematológico e bioquímico de éguas da raça Mangalarga Marchador no período de transição mantidas a pasto

The present study aimed to evaluate the effects of the transition period on hematological and biochemical constituents in Mangalarga Marchador mares. Forty-eight mares were used to form a maintenance group (MG) and transition group (TG), formed by pregnant mares and, after delivery, infants. Blood samples were collected at the following times: T-60 (60 d pre-delivery), T-30 (30 d pre-delivery), T-15 (15 d pre-delivery), T0 (first 6h post-delivery), T15 (15 d post-delivery), T30 (30 d post-delivery), and T60 (60 d post-delivery). The TG had lower values (P< 0.05) of red blood cells, hematocrit and hemoglobin at T0, T15, T30 and T60 times than MG. The mean corpuscular volume was lower in MG (P< 0.05) than in TG (T0, T15, T30 and T60) and mean corpuscular hemoglobin concentration was higher (P< 0.05) in MG than in TG (T15, T30 and T60). On the other hand, the diameter distribution of red blood cells presented a lower value (P< 0.05) in MG than in TG (T15 and T30). Mares in transition period presented regenerative anemia. The results demonstrate physiological metabolic variations of different intensities during pregnancy, delivery and early lactation.(AU)

O presente estudo teve como objetivo avaliar os efeitos do período de transição em constituintes hematológicos e bioquímicos em éguas Mangalarga Marchador. Foram utilizadas 48 éguas para formar um grupo de manutenção (GM) e um grupo de transição (GT), composto por éguas gestantes e, após o parto, lactentes. Amostras de sangue foram coletadas nos seguintes tempos: T-60 (60 dias pré-parto), T-30 (30 dias pré-parto), T-15 (15 dias pré-parto), T0 (seis primeiras horas pós-parto), T15 (15 dias pós-parto), T30 (30 dias pós-parto) e T60 (60 dias pós-parto). O GT apresentou valores menores (P<0,05) de hemácias, hematócrito e hemoglobina, nos tempos T0, T15, T30 e T60, do que o GM. O volume corpuscular médio foi menor no GM (P<0,05) do que no GT (T0, T15, T30, T60) e a concentração corpórea de hemoglobina corpórea foi maior (P<0,05) no GM do que no GT (T15, T30, T60). Por outro lado, a distribuição do diâmetro dos eritrócitos apresentou um valor menor (P<0,05) no GM do que no GT (T15 e T30). Éguas em período de transição apresentam anemia regenerativa. Os resultados demonstram variações metabólicas de diferentes intensidades durante a gestação, o parto e o início de lactação.(AU)

Padronização de uma PCR para diagnóstico molecular de Microsporum canis em amostras de pelos e crostas de cães e gatos / Standardization of a pcr for molecular diagnosis of microsporum canis in samples of fur and crusts of dogs and cats

Objetivou-se neste estudo padronizar um protocolo de reação em cadeia da polimerase (PCR) para detecção de Microsporum canis em amostras de pelos e/ou crostas de cães e gatos. Foram selecionadas 48 amostras previamente identificadas por meio de cultura. Destas, 23 foram positivas para dermatófitos no cultivo. Padronizou-se a PCR a partir de primers desenhados para o alvo M. canis. Sessenta e um por cento (14/23) das amostras positivas para dermatófitos foram identificadas como M. canis em cultura. Desse total, 71,4% (10/14) apresentaram um fragmento de 218pb compatível com o esperado para a espécie fúngica alvo dessa reação. Observou-se uma sensibilidade de 71,4% e especificidade de 100% na PCR, além de uma boa concordância entre essas técnicas de diagnóstico (Kappa: 0,78; P<0,0001). O protocolo utilizado neste estudo apresentou alta especificidade na detecção de M. canis diretamente de amostras de pelos e/ou crostas de cães e gatos, viabilizando um diagnóstico mais rápido e específico, podendo esse protocolo ser empregado como um método confirmatório para agilizar a detecção de M. canis.(AU)

The aim of this study was to standardize a Polymerase Chain Reaction protocol (PCR) for the detection of Microsporum canis in fur and/or crusts of dogs and cats. 48 samples previously identified by culture were selected. Of these, 23 were positive for dermatophytes in culture. PCR was standardized from drawn primers whose target is M. canis. A total of 61% (14/23) of the dermatophyte positive samples were identified as M. canis in culture. Of this total, 71.4% (10/14) presented a fragment of 218bp compatible with that expected for the fungal species target of the reaction. A sensitivity of 71.4% and specificity of 100% in the PCR were observed, in addition to a good agreement between the techniques (Kappa: 0.78; P<0.0001). The protocol used in this study showed high specificity in the detection of M. canis directly from fur and/or crusts of dogs and cats, making possible a faster and more specific diagnosis. This protocol could be used as a confirmatory method, speeding the detection of M. canis.(AU)

Hypotonic enteral electrolyte solutions administered by nasoesophageal tube in continuous flow in dogs dehydrated by water restriction: Part 1 / Soluções eletrolíticas enterais hipotônicas administradas por sonda nasoesofágica em fluxo contínuo em cães desidratados por restrição hídrica: Parte 1

The present study assessed and compared the effects of hypotonic enteral electrolyte solutions administered by nasoesophageal tube in continuous flow in dogs submitted to water restriction on packed cell volume; total serum protein and serum osmolarity concentrations; blood volume; plasma glucose and lactate levels; blood gas analysis, anion gap, and strong ion difference. Six adult dogs were used (four males and two females). All animals were submitted to both proposed treatments in a crossover design 6×2. The treatments were as follows: ESmalt consisting of 5g sodium chloride, 1g potassium chloride, 1g calcium acetate, 0.2g magnesium pidolate, and 9.6g maltodextrin that were diluted in 1.000mL water (measured osmotic concentration of 215mOsm L−1) and ESdext consisting of 5g sodium chloride, 1g potassium chloride, 1g calcium acetate, 0.2g magnesium pidolate, and 9.6g dextrose that were diluted in 1.000mL water (measured osmotic concentration of 243mOsm L−1). All solutions were administered at 15ml kg−1 h−1 for 4 hours. Both solutions increased the plasma volume in dehydrated dogs without causing adverse effects. However, ESmalt was more effective in promoting the increase in blood volume.(AU)

O presente estudo avaliou e comparou os efeitos de soluções eletrolíticas enterais hipotônicas, administradas por sonda nasoesofágica em fluxo contínuo em cães submetidos a restrição hídrica, sobre o hematócrito, proteínas totais séricas, osmolaridade sérica, volemia, glicose e lactato plasmáticos, hemogasometria, ânion gap e DIF. Foram utilizados seis cães adultos (quatro machos e duas fêmeas). Todos os animais foram submetidos aos dois tratamentos propostos, em um delineamento crossover 6×2. Os tratamentos foram os seguintes: SEmalt - 5g de cloreto de sódio, 1g de cloreto de potássio, 1g de acetato de cálcio, 0,2g de pidolato de magnésio e 9,6g de maltodextrina, diluídos em 1.000mL de água (osmolaridade mensurada: 215mOsm L -1 ); SEdext - 5g de cloreto de sódio, 1g de cloreto de potássio, 1g de acetato de cálcio, 0,2g de pidolato de magnésio e 9,6g de dextrose, diluídos em 1.000mL de água (osmolaridade mensurada: 243mOsm L -1 ). Todas as soluções foram administradas no volume de 15mL kg -1 hora -1 , durante quatro horas, em fluxo contínuo. Ambas as soluções aumentaram o volume plasmático em cães desidratados, sem gerar o aparecimento de efeitos adversos. Porém, a SEmalt foi mais eficaz em promover a expansão da volemia.

Imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas / Immunodetection of Toxoplasma gondii in placenta tissue of naturally infected goats

A imuno-histoquímica (IHQ) é considerada uma ferramenta rápida e precisa para a identificação de protozoários, como Toxoplasma gondii, em tecidos fetais e placentários. Neste estudo foi avaliada a imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas. Foram coletadas e analisadas 80 amostras de placentas de cabras procedentes de único rebanho com sorologia positiva para T. gondii na técnica de ELISA. Na histopatologia, 27/80 amostras apresentaram lesões sugestivas de infecção por protozoários. Após a avaliação histopatológica, procedeu-se à realização da técnica de imuno-histoquímica, obtendo-se 85,2% (23/27) de amostras com marcação positiva. A imunodetecção ocorreu no epitélio de revestimento das vilosidades coriônicas e foi classificada de acordo com o grau de intensidade da imunomarcação. Também foi evidenciada imunomarcação no interior dos vasos sanguíneos fetais em 8,69% (2/23) das amostras. Este estudo demonstrou que a técnica de IHQ se comportou como uma ferramenta valiosa no diagnóstico da infeção por T. gondii em tecido placentário de cabras naturalmente infectadas e complementou, de forma decisiva, o diagnóstico, além de agregar maior valor aos resultados obtidos nas análises histopatológica e sorológica.(AU)

Immunohistochemistry (IHC) is considered to be a rapid and accurate tool for the identification of protozoa such as Toxoplasma gondii in fetal and placental tissues. In this study, we evaluated the immunodetection of Toxoplasma gondii in placental tissue from naturally infected goats. A total of 80 samples of goat placentas from a single herd with positive ELISA serology for T. gondii were collected and analyzed. In the histopathology, 27/80 samples presented lesions suggestive of protozoal infection. After the histopathological evaluation, the immunohistochemistry technique was performed, obtaining 85.2% (23/27) of samples with positive marking. Immunodetection occurred in the lining epithelium of the chorionic villi and was classified according to the degree of intensity of the immunostaining. Immunostaining within the fetal blood vessels was also evidenced in 8.69% (2/23) of the samples. This study demonstrated that the IHQ technique behaved as a valuable tool in the diagnosis of T. gondii infection in placental tissue of naturally infected goats completing the diagnosis in a decisive way and adding greater value to the results obtained in the histopathological and serological analysis.(AU)

Preweaning injections of bovine ST enhanced reproductive performance of Bos indicus-influenced replacement beef heifers.

A 3 yr study evaluated the effects of three preweaning injections of bovine ST, administered 14 d apart, on growth and reproductive performance of Bos indicus-influenced beef heifers. On d 0 of each year, suckling Angus × Brangus heifers (n = 15 heifers/treatment/yr) were stratified by BW (147 ± 20 kg) and age (134 ± 11 d) and randomly assigned to receive an s.c. injection of saline (SAL; 5 mL; 0.9% NaCl) or 250 mg of sometribove zinc (BST; Posilac, Elanco, Greenfield, IN) on d 0, 14, and 28. Heifers and respective dams were managed as a single group on bahiagrass (Paspalum notatum) pastures from d 0 until weaning (d 127). From d 127 to 346, heifers were grouped by treatment, allocated to bahiagrass pastures (1 pasture/treatment/yr) and fed a molasses-based supplement (2.9 kg/heifer daily; DM basis) until d 346. Blood samples were collected on d 0, 14, 28, 42, and then every 9-10 d from d 179 to 346. In yr 3, liver biopsy samples were collected on d 0, 42, and 263. Heifers were exposed to mature Angus bulls from d 263 to 346. Growth performance and physiological parameters were analyzed using the MIXED procedure, whereas reproductive variables were analyzed using the GLIMMIX procedure of SAS. Effects of treatment × year and treatment × year × time were not detected for any variable measured in this study (P ≥ 0.14), except for calving percentage (P = 0.03). Heifers assigned to BST injections had greater overall plasma concentrations of IGF-1 and ADG from d 0 to 42 (P ≤ 0.05), less ADG from d 42 to 127 (P = 0.04), but had similar BW at weaning and postweaning ADG (P ≥ 0.25) compared to SAL heifers. Heifers assigned to BST tended to achieve puberty 26 d earlier (P = 0.10), had greater percentage of pubertal heifers on d 244, 263, 284, and 296 (P ≤ 0.04), tended to have greater overall pregnancy percentage (P = 0.10), and had greater (P ≤ 0.05) calving percentages in yr 1 and 2 (but not yr 3; P = 0.68) compared to SAL heifers. Liver mRNA expression of GHR-1B and IGF-1 on d 0 and 42 did not differ between treatments (P ≥ 0.15), but was greater for BST vs. SAL heifers on d 263 (P ≤ 0.02). Hence, administering three injections containing 250 mg of sometribove zinc at 14 d intervals before weaning (between 135 and 163 d of age) induced long-term impacts on liver gene expression and may be a feasible management practice to enhance puberty and pregnancy attainment in B. indicus-influenced replacement beef heifers.

Occurrence of Mycoplasma bovigenitalium and Ureaplasma diversum in dairy cattle from to Pernambuco state, Brazil / Ocorrência de Mycoplasma bovigenitalium e Ureaplasma diversum em bovinos leiteiros do estado de Pernambuco, Brasil

The objective of this study was to conduct an investigation of Mycoplasma bovigenitalium and Ureaplasma diversum infections in cattle in the microregion of the Ipanema Valley, state of Pernambuco, Brazil. Vaginal swabs were collected from 355 breeding cows in reproductive age and were analyzed by multiplex PCR (mPCR) and culture. An epidemiological investigation of risk factors was performed for Mollicutes. mPCR analysis showed that, 9.29% (33/355) of the cows were positive for M. bovigenitalium and 21.69% (77/355) for U. diversum; coinfection was observed in 2.81% (10/355) of the cows. The microbiological isolation showed, 81.81% (27/33) of Mycoplasma spp. and 24.67% (19/77) of Ureaplasma spp.. The risk factors related to Mollicutes infection identified were semi-intensive breeding system (OR= 4.6), pasture rent (OR= 3.6), non-isolation of animals with reproductive disorders (OR= 3.2), and natural mounting and artificial insemination (OR= 3.5). There was a significant association between Mollicutes infection and abortions in the first gestational third (P= 0.001). This is the first record of M. bovigenitalium and U. diversum infection in cows in the semiarid region of the state of Pernambuco, Brazil. Preventive measures directed to the identified risk factors can decrease the occurrence of Mollicutes in these herds.(AU)

O objetivo deste estudo foi realizar uma investigação de Mycoplasma bovigenitalium e Ureaplasma diversum em bovinos leiteiros da microrregião do Vale do Ipanema, estado de Pernambuco, Brasil. Foram coletados suabes vaginais de 355 vacas em idade reprodutiva. As amostras foram analisadas por multiplex PCR (mPCR) e cultura. Foi realizada uma investigação dos fatores de risco para Mollicutes. Na mPCR, 9,29% (33/355) das vacas foram positivas para M. bovigenitalium e 21,69% (77/355) para U. diversum; coinfecção foi observada em 2,81% (10/355) das vacas. O isolamento microbiológico mostrou crescimento de Mycoplasma spp. em 81,81% (27/33) das amostras e em 24,67% (19/77) para Ureaplasma spp. Os fatores de risco relacionados à infecção por Mollicutes identificados foram sistema de produção semi-intensivo (OR= 4,6), aluguel de pastagem (OR= 3,6), não isolamento de animais com desordens reprodutivas (OR= 3,2) e monta natural e inseminação artificial (OR= 3,5). Houve uma associação significativa entre a infecção por Mollicutes e abortos no primeiro terço gestacional (P=0,001). Este é o primeiro relato da infecção por M. bovigenitalium e U. diversum em vacas na região semiárida do estado de Pernambuco, Brasil. As medidas preventivas direcionadas aos fatores de risco identificados podem diminuir a ocorrência de Mollicutes nesses rebanhos.(AU)

Anti-Müllerian hormone reduces growth rate without altering follicular survival in isolated caprine preantral follicles cultured in vitro.

The aim of the present study was to evaluate the effect of anti-Müllerian hormone (AMH), with and without FSH, on the in vitro development of isolated caprine preantral follicles, as well as follicular steroid production and mRNA levels of AMH, hormone receptors (AMH and FSH), CYP19A1 (cytochrome P450, family 19, subfamily A, polypeptide 1), CYP17 (cytochrome P450, family 17, subfamily A, polypeptide 1), HSD3B (3-beta-hydroxysteroid dehydrogenase) and Myc (myelocytomatosis oncogene). Isolated secondary follicles were cultured in minimum essential medium alpha (α-MEM+) alone or supplemented with 50ng mL-1 AMH and/or 100ng mL-1 FSH added sequentially on different days of culture. Follicles were cultured for a total of 18 days, with different media during the first (Days 0-9) and second (Days 10-18) halves of the culture period, resulting in six treatment groups, as follows: α-MEM+/α-MEM+, FSH/FSH, AMH/AMH, AMH+FSH/AMH+FSH, AMH/FSH, and FSH/AMH. Follicle development was evaluated on the basis of follicular growth, oocyte maturation and steroid secretion. There was a decrease in follicular growth rate in the AMH, AMH+FSH and AMH/FSH treatment groups compared with α-MEM+ and FSH treatment groups (P<0.05). However, the different culture conditions had no effect on rates of meiotic resumption and steroid secretion (P>0.05). Moreover, follicles cultured in the presence of FSH had lower levels of AMH receptor type II (AMHRII) mRNA compared with non-cultured control (freshly isolated follicles), and the AMH and AMH/FSH treatment groups. In conclusion, AMH reduces the follicular growth rate of isolated goat preantral follicles in vitro without affecting follicular survival.

Achatina fulica (Gastropoda: Pulmonata): Occurrence, environmental aspects and presence of nematodes in Sergipe, Brazil

Abstract The objective of this study was to identify the occurrence of Achatina fulica in the Great Aracaju-SE Territory. This research was developed during the months of June 2016 to May 2017. The plots methodology was used for the monthly collections, following the collection time of 20 minutes per area. The mollusks were measured and soon afterwards the parasitological analysis of the mucus and faeces was performed. Molecular identification was performed to evaluate the possible presence of Angiostrongylus sp. We observed that the occurrence of A. fulica was more representative in urban areas, especially in vacant lots with the presence of litter and decomposing materials. In the specimens of A. fulica examined were male and female nematodes of the genus Rhabditis, in the research municipalities, except in Barra dos Coqueiros. No parasites of the genus Angiostrongylus were found in the mollusks examined. The influence of the presence of litter on the plots associated with the frequency of A. fulica was statistically significant. There was a correlation between the influence of rainfall and the frequency of A. fulica. A significant relationship between moisture and frequency and nematode positivity was identified. It was also observed that the larger the mollusk, the greater the chances of being infected by nematodes. Scientific knowledge about the biology of the A. fulica species associated with human intervention such as the maintenance of land and adequate sanitary measures may be important factors for a better control of this invasive mollusk.

Resumo O objetivo deste estudo foi verificar a ocorrência de A. fulica no território da Grande Aracaju, Sergipe, avaliando a infecção destes moluscos por nematódeos, a influência das condições climáticas e das características do ambiente. Esta pesquisa foi desenvolvida durante os meses de junho de 2016 a maio de 2017. Foi utilizada a metodologia de parcelas para as coletas mensais de 20 min por área, um total de 15 parcelas distribuídas em 5 municípios. Os moluscos foram mensurados e logo em seguida realizada a análise parasitológica do muco e fezes. Foi realizada a identificação molecular para avaliar a possível presença de Angiostrongylus sp. Observamos que a ocorrência de A. fulica foi mais representativa em áreas urbanas, sobretudo em terrenos baldios com a presença de lixo e materiais em decomposição. Nos exemplares de A. fulica examinados foram encontrados nematódeos do gênero Rhabditis, nos municípios da pesquisa, exceto em Barra dos Coqueiros. Não foram encontrados parasitos do gênero Angiostrongylus nos moluscos examinados. A influência da presença de lixo nas parcelas associado à frequência da A. fulica mostrou-se estatisticamente significativa. Houve correlação entre a influência da chuva e a frequência de A. fulica. Foi identificada uma relação significativa entre a umidade e a frequência e a positividade para nematódeos. Foi também observado que quanto maior o molusco, maiores são as chances deste apresentar nematódeos. O conhecimento científico a respeito da biologia da espécie A. fulica associado à intervenção humana como, a manutenção de terrenos e medidas sanitárias adequadas podem ser fatores importantes para um controle malacológico mais adequado para esta espécie invasora.

Impact of insulin concentration and mode of FSH addition on the in vitro survival and development of isolated bovine preantral follicles.

UNLABELLED: The insulin and FSH are two important substances in the folliculogenesis process. Thus, the hypothesis of this experiment is that insulin concentration and the form of FSH addition affect the in vitro survival, growth, and estradiol production after culture of isolated bovine preantral follicles. The effects of insulin concentration (experiment 1) and the influence of both fixed and sequential concentrations of FSH (experiment 2) on the in vitro survival and development of bovine preantral follicles were investigated in this study by IVC for 18 days. In experiment 1, on Day 18 of culture, the addition of insulin at all concentrations promoted follicular survival rates significantly higher than that of the control, with the 10-ng/mL insulin treatment showing values significantly higher than the other treatments. The addition of 5- and 10-ng/mL insulin promoted higher follicular growth than the control and other treatments. In experiment 2, FSH 100 had a higher percentage of follicular viability compared with the control. FSH 100 produced follicle diameters significantly higher than those of the control and FSH seq. TREATMENT: Estradiol levels in the presence of FSH (fixed concentration) were significantly higher than the other treatments. In conclusion, the association of insulin (10 ng/mL) and fixed concentration FSH (100 ng/mL) provides high rates of survival, growth, and estradiol production in bovine preantral follicles.

Impact of growth hormone (GH) and follicle stimulating hormone (FSH) on in vitro canine preantral follicle development and estradiol production.

OBJECTIVE: Evaluate the effect of different concentrations of growth hormone (GH) on the in vitro development of domestic dog (Canis lupus familiaris) preantral follicles in the presence or absence of follicle stimulating hormone (FSH). METHODS: Secondary preantral follicles, isolated by microdissection, were cultured in a medium composed of αMEM with bovine serum albumin (BSA), glutamine, hypoxanthine, insulin, transferrin, selenium and ascorbic acid (αMEM(+)-control) added at different concentrations of GH (GH10 ng/ml or GH50 ng/ml) and FSH (GH10+FSH, GH50+FSH). Follicle development was evaluated based on the percentage of intact follicles, antrum formation, follicular diameter, follicular viability using fluorescent markers and estradiol production. RESULTS: GH50 was the only treatment that maintained the same percentage of normal morphologically follicles from day 0 to day 18 of culture (P<0.05). For all treatments, except the control, follicles were viable throughout the 18 days of culture (P<0.05). GH50 supplemented with FSH (GH50+FSH) resulted in the highest average follicular diameter (P<0.05) from day 12 to 18. Follicles from both the control and the GH50+FSH treatment groups actively and increasingly secreted estradiol from day 6 to 18 of culture (P<0.05). CONCLUSIONS: Our study demonstrates that GH benefits the maintenance of follicular morphology in a dose-dependent manner and, in association with FSH, stimulates in vitro follicular growth and estradiol production.

In vitro development of secondary follicles from pre-pubertal and adult goats cultured in two-dimensional or three-dimensional systems.

The aim of this study was to evaluate the influence of two-dimensional (2D) and three-dimensional (3D) alginate culture systems on in vitro development of pre-antral caprine follicles. In addition, the influence of the reproductive age of the ovary donor on the in vitro culture success was investigated. Pre-antral follicles from pre-pubertal or adult goats were isolated and cultured directly on a plastic surface (2D) or encapsulated in an alginate-based matrix (3D). After 18 days, the oocytes underwent in vitro maturation (IVM) and in vitro fertilization (IVF) to produce embryos. The 3D system showed higher rates of follicle survival, lower rates of oocyte extrusion, and a greater number of recovered oocytes for IVM and IVF (P < 0.05). Only pre-antral follicles from adult animals produced MII oocytes and embryos. The estradiol concentrations increased from day 2 to day 12 of culture in all groups tested (P < 0.05). Conversely, progesterone concentrations were lower in 3D-cultured follicles than in 2D-cultured follicles, with differences on days 2 and 6 of culture (P < 0.05). We provide compelling evidence that a 2D or 3D alginate in vitro culture system offers a promising approach to achieving full in vitro development of caprine pre-antral follicles to produce mature oocytes that are capable of fertilization and viable embryos.

Chirality of weakly bound complexes: the potential energy surfaces for the hydrogen-peroxide-noble-gas interactions.

We consider the analytical representation of the potential energy surfaces of relevance for the intermolecular dynamics of weakly bound complexes of chiral molecules. In this paper we study the H2O2-Ng (Ng=He, Ne, Ar, Kr, and Xe) systems providing the radial and the angular dependence of the potential energy surface on the relative position of the Ng atom. We accomplish this by introducing an analytical representation which is able to fit the ab initio energies of these complexes in a wide range of geometries. Our analysis sheds light on the role that the enantiomeric forms and the symmetry of the H2O2 molecule play on the resulting barriers and equilibrium geometries. The proposed theoretical framework is useful to study the dynamics of the H2O2 molecule, or other systems involving O-O and S-S bonds, interacting by non-covalent forces with atoms or molecules and to understand how the relative orientation of the O-H bonds changes along collisional events that may lead to a hydrogen bond formation or even to selectivity in chemical reactions.

Potencial antioxidante in vitro das folhas da Bauhinia ungulata L / Antioxidant Potential in vitro of leaves of Bauhinia ungulata L

Bauhinia ungulata L. (Pata de Vaca) is native to Brazil, mainly found in the Amazon, Cerrado and Atlantic Forest, and popularly used in the treatment of diabetes. This paper describes an analysis of the in vitro antioxidant potential of crude ethanol extract and fractions (hexane, chloroform, ethyl acetate and hydroalcoholic residue) obtained from leaves of this species, using three different methods: 1) scavenging of organic radical 2,2 - diphenyl-1-picrylhydrazyl(DPPH•), 2) reducing phosphomolybdenum complex and 3) assaying the products formed during lipid peroxidation (TBARS). The antioxidant potential of the extract and fractions varied with the methodology, but the ethyl acetate fraction showed activity similar to the standards used in the three methods. The results provide evidence that the leaves of B. ungulata L. area potential source of natural antioxidants, stimulating new studies aimed at the production of potential drugs for the treatment of diseases related to free radicals...

A Bauhinia ungulata L. (Pata de Vaca) é uma planta nativa do Brasil, encontrada principalmente na região Amazônica, Cerrado e Mata Atlântica, utilizada popularmente no tratamento do diabetes. Este trabalho descreve uma análise in vitro do potencial antioxidante do extrato bruto etanólico e frações (hexano, clorofórmio, acetato de etila e hidroalcoólica residual) obtidos de folhas desta espécie, através de três metodologias distintas: 1) captura de radical orgânico 2,2-difenil-1-picril-hidrazila-DPPH•, 2) redução do complexo fosfomolibdênio e 3) quantificação de produtos formados durante a peroxidação de lipídios (TBARS). O potencial antioxidante do extrato e frações foi variável de acordo com a metodologia utilizada com destaque para a fração acetato de etila que apresentou atividade similar aos padrões utilizados nos três métodos. Os resultados obtidos fornecem evidências de que as folhas de B. ungulata L. são uma potencial fonte de antioxidantes naturais, estimulando novos estudos que objetivem a produção de possíveis medicamentos destinados ao tratamento de patologias associadas aos radicais livres...

Effect of morphological integrity, period, and type of culture system on the in vitro development of isolated caprine preantral follicles.

The aims of this study were the following: (1) to define an optimal period for the IVC of isolated caprine preantral follicles, (2) to verify the relationship between follicular morphology (intact, extruded, and degenerate follicles) and estradiol production, and (3) to evaluate the effects of the bidimensional (2D) and three-dimensional (3D) culture systems on the in vitro development of caprine preantral follicles. Three experiments were performed. In experiments 1 and 2, the isolated secondary follicles were cultured for 18, 24, and 30 days or 30, 36, and 42 days, respectively. In experiment 3, the optimal culture period from experiment 2 was used for 2D and 3D culture systems. After culture, the oocytes were submitted to IVM. The morphological integrity, antral cavity formation rates, follicular diameter, presence of healthy, grown oocytes (≥110 µm), rates of resumption of meiosis, and estradiol concentrations were evaluated. In experiment 1, the percentage of oocytes that resumed meiosis was higher in oocytes cultured for 30 days (48.84%) than in oocytes cultured for 18 and 24 days (15% and 20.93%, respectively). In experiment 2, the percentage of oocytes that resumed meiosis was significantly higher in oocytes cultured for 30 and 36 days (47.5% and 50%, respectively) than in oocytes cultured for 42 days (20%). The estradiol concentrations on Day 12 of culture were similar for normal and extruded follicles and higher than those observed in degenerate follicles at the end of the culture period. In conclusion, the 36-day culture period resulted in the highest rates of meiosis resumption. In addition, because the loss of follicular integrity affects the patterns of estradiol production, follicular integrity is a good predictor of follicular quality.

Radon exposure at a radioactive waste storage facility.

The Waste Management Department of Nuclear and Energy Research Institute (IPEN) is responsible for the safety management of the waste generated at all internal research centers and that of other waste producers such as industry, medical facilities, and universities in Brazil. These waste materials, after treatment, are placed in an interim storage facility. Among them are (226)Ra needles used in radiotherapy, siliceous cake arising from conversion processes, and several other classes of waste from the nuclear fuel cycle, which contain Ra-226 producing (222)Rn gas daughter.In order to estimate the effective dose for workers due to radon inhalation, the radon concentration at the storage facility has been assessed within this study. Radon measurements have been carried out through the passive method with solid-state nuclear track detectors (CR-39) over a period of nine months, changing detectors every month in order to determine the long-term average levels of indoor radon concentrations. The radon concentration results, covering the period from June 2012 to March 2013, varied from 0.55 ± 0.05 to 5.19 ± 0.45 kBq m(-3). The effective dose due to (222)Rn inhalation was further assessed following ICRP Publication 65.

Estudo de biocompatibilidade in vivo e in vitro do extrato hidroalcoólico de Cissus sicyoides L. - Vitaceae em ratos / Study of in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts in rats

Biocompatibilidade é a capacidade de um material exercer funções específicas quando aplicado em contato com tecidos vivos de determinado hospedeiro, sem, contudo, causar danos ou prejuízo ao mesmo. Este trabalho objetivou determinar a biocompatibilidade in vivo e in vitro do extrato hidroalcoólico do Cissus sicyoides L - Vitaceae. Foram utilizados 30 ratos (Rattus novergicus albinus wistar), com idade entre 45 e 90 dias e pesando entre 170 e 260 g. Os animais foram divididos em 3 grupos (A1, A2 e A3) de 6 animais cada para o teste in vivo, os quais foram sacrificados com 2, 4 e 6 dias, respectivamente. Para o teste in vitro, foram utilizados 12 animais para obtenção do índice de aderência e da capacidade fagocítica dos macrófagos de ratos do grupo controle e do grupo experimental. Nos resultados encontrados no teste in vivo, conclui-se que o extrato apresentou-se biocompatível, visto que não provocou alterações significativas no tecido. Já no teste in vitro, o mesmo não se apresentou biocompatível, pois o extrato puro apresentou índice de aderência baixo (7,1) e taxa de fagocitose elevada (35,7), indicando diferença significante quando comparado ao controle. Porém, quando diluído, o extrato se mostrou inócuo, devido ao aumento dos valores do índice de aderência nas concentrações de 1/10 (61,4) e 1/100 (74,3) nos ensaios, as quais não apresentaram diferença significante quando comparadas ao controle. Após a análise dos dados, concluiu-se que a solução diluída do extrato hidroalcoólico do Cissus sicyoides L. não causa danos ou prejuízos. Entretanto, como nem todos os efeitos farmacológicos foram testados no presente trabalho, não se pode inferir automaticamente que ele é biocompatível em todos os casos.

Biocompatibility is the ability of a material to perform specifictasks when applied to living tissues without causing damage or injuries to it. Thus, this study aimed at determining the in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts. A total of 30 rats (Rattus norvegicus Albinus Wistar), with ages ranging from 45 to 90 days and weighing between 170 and 260g were used. The animals were divided into 3 groups (A1, A2 and A3) with 6 animals each, for the in vivo test, which were sacrificed after 2, 4 and 6 days, respectively. For in vitro test, 12 animals were used to obtain the index of adherence and phagocytic ability of macrophages of rats from the control and the experimental groups. In results found for the in vivo test, it was concluded that the extract was biocompatible, whereas no significant changes were observed in the tissue. As to the in vitro test, the extract was not biocompatible, since the pure extract showed a low rate of adherence (7.1) and a high rate of phagocytosis (35.7), indicating a significant difference when compared to the control group. However, when diluted, the extract was shown to be harmless, due to an increase in the values of the adherence index at the following concentrations : 1/10 (61.4) and 1/100 (74.3) in the tests, which showed no significant differences when compared to the control group. After analyzing the data, it was concluded that since the infusion of the plant is a kind of dilution, its use does not cause any harm to the body. A new study is necessary at the moment to possibily demonstrate its effects on the long term.

Vascular endothelial growth factor-A(165) (VEGF-A(165)) stimulates the in vitro development and oocyte competence of goat preantral follicles.

The aim of this study was to evaluate the effect of vascular endothelial growth factor-A(165) (VEGF-A(165)) on the in vitro development of goat secondary preantral follicles. Preantral follicles (≥150 µm in diameter) were isolated from the ovaries of adult mixed-breed goats and individually cultured for 18 days in αMEM in the absence (control) or presence of VEGF-A(165) at concentrations of 10 ng/ml (VEGF10) and 100 ng/ml (VEGF100). Analyses of follicular survival, diameter, antrum formation and rate of daily growth were performed every 6 days. At the end of the culture period, morphologically normal oocytes (≥110 µm in diameter) were taken for in vitro maturation (IVM). The results demonstrated that all follicles presented oocytes and granulosa cells that were morphologically normal and after labeling with calcein-AM, high rates of oocyte viability were observed in all treatments. The follicular diameter and the growth rate achieved in the presence of VEGF10 were higher than those of the control. Both treatments with VEGF-A(165) showed higher rates of oocyte recovery for IVM when compared with the control. Moreover, only the addition of VEGF-A(165) permitted oocytes grown in vitro to reach metaphase II. Thus, the addition of VEGF-A(165) to the culture medium improves the development of goat preantral follicles cultured in vitro, allowing the production of mature oocytes.

Efeito do nível de ácido linoléico na ração de matrizes pesadas sobre o peso, composição e eclosão dos ovos / Effect of linoleic acid level in the diets on the weight, composition and eclodibility of broiler breeders eggs

Avaliaram-se os efeitos dos níveis de ácido linoléico na dieta de matrizes pesadas, de 27 a 40 semanas de idade, sobre o peso e composição dos ovos, porcentagem de eclosão, peso do pinto, porcentagem de saco vitelino em relação ao peso do pinto e composição de ácidos graxos da gema. Os tratamentos foram definidos pelo nível de ácido linoléico na dieta. O peso dos ovos e a composição de ácidos linoléico na gema foram mais altos com a inclusão de 1,93 por cento de ácido linoléico que a de 1,48 por cento (59,5g vs 59,0g e 19,5 por cento vs 15,5 por cento, respectivamente). Não houve diferença entre os tratamentos quanto às porcentagens de gema, albúmen, casca, saco vitelino, eclosão e peso do pinto.

The effect of diet linoleic acid level on egg weight, egg composition, hatchability, chick weight, yolk sac percentage relative to chick weight and yolk fatty acid composition of broiler breeders Ross 305, from 27 to 40 weeks of age, were evaluated using two diets with two levels of linoleic acid. Egg weight and linoleic acid yolk composition were higher for diet with 1.93 percent of linoleic acid than for diet with 1.48 percent (59.5g vs 59.0g and 19.5 percent vs 15.5 percent, respectively). No linoleic acid effects on yolk, albumen, egg shell, yolk sac percentage and hatchability and chick weight were observed.

The effect of diverticular disease on the colonic J pouch.

OBJECTIVE: The aim of this study was to assess the impact of the diverticular disease (DD) on function and on postoperative complications of the colonic J-pouch (CJP) with pouch-anal anastomosis. METHODS: Patients who underwent a CJP between December 1990 and August 2001, were retrospectively reviewed. The presence of DD in the CJP was assessed on pouchogram prior to ileostomy closure. A questionnaire designed to evaluate the degree of continence (total incontinence score (IS): 0 = worst, 20 = best) and pouch evacuation (total evacuation score (ES): 0 = worst, 28 = best) was used for comparison between patients with DD and those without DD (NDD). RESULTS: Sixty-six patients (47 males; 19 females) with a median age of 68 years (range 28-87 years) were included. The median follow-up period was 22 months (range 2-106 months). Twenty-four patients comprised the DD group and 42 were in the NDD group. The two groups were comparable for age, gender and time from ileostomy closure; all patients with postoperative chemoradiation therapy were in the NDD group. The total ES and IS total did not significantly differ between the two groups with a P-value of 0.11 and 0.09 respectively. Furthermore, there was no significant difference in the total incidence of pouch complications between the two groups (3 strictures, 1 leak, 1 fistula in the NDD group vs. 1 pelvic sepsis in the DD group; P = 0.4). CONCLUSIONS: The presence of DD in a CJP does not seem to impact pouch function or the postoperative complication rate.