Orientador: Arielle Cristina Arena / Banca: Daniela Cristina Ceccatto Gerardin / Banca: Carla Dal Bianco Fernandez / Banca: Luis Antonio Justulin Junior / Banca: Luis Gustavo de Almeida Chuffa / Banca: Cândida Aparecida Leite Kassuya / Banca: Patrícia Fernanda Felipe Pinheiro / Resumo: Fipronil é um inseticida amplamente utilizado na agricultura, medicina veterinária e saúde pública, e recentemente tem sido listado como um provável desregulador endócrino. Estudos demonstram que este composto pode afetar a função da tireoide, parâmetros reprodutivos e o sistema nervoso central. No entanto, a maioria dos estudos realizados com o fipronil avaliou sua toxicidade aguda, e poucos se referem aos seus possíveis efeitos reprodutivos, especialmente se a exposição ocorrer durante a gestação e lactação. Este estudo objetivou avaliar os efeitos resultantes da exposição perinatal ao fipronil e suas possíveis repercussões tardias sobre parâmetros reprodutivos masculinos e femininos, em ratos. Ratas prenhes foram expostas a três doses de fipronil (0,03; 0,3 ou 3 mg/kg) do dia gestacional 15 até o dia pós-natal 7, por gavage. Durante o tratamento, foram monitorados o consumo de água e ração e o peso corporal das ratas expostas. Após o desmame, foram coletados o sangue das mães para análises bioquímicas e órgãos para registro do peso. A prole masculina foi avaliada através dos seguintes parâmetros: peso corporal, distância anogenital e idades da separação prepucial e da descida testicular. Na vida adulta, peso de órgãos reprodutores, contagens, morfologia e motilidade espermática, histo-morfometria do testículo e epidídimo, contagem de células de Sertoli, comportamento e preferência sexual, fertilidade e expressão de receptores de andrógenos no testículo foram investigados. Nos filhotes do sexo feminino foram avaliados a distância anogenital, idade de instalação de puberdade, ciclo estral, dosagem hormonal, fertilidade, comportamento sexual e histologia de útero e ovário. O tratamento não alterou os parâmetros bioquímicos e os pesos da maioria dos órgãos analisados das mães expostas ao fipronil, com exceção do peso da hipófise, que apresentou uma redução no grupo exposto a menor dose... / Abstract: Fipronil is an insecticide widely used in agriculture, veterinary medicine and public health, and recently has been listed as a possible endocrine disruptor. Studies demonstrate that this compound can affect the thyroid function, reproductive parameters and central nervous system. However, most of studies performed with fipronil evaluated the acute toxicity, and a few of them refer to its reproductive effects, especially if the exposure occurs during gestation and lactation. This study aimed to evaluate the effects resulting from perinatal exposure to fipronil and its possible late repercussion on male and female reproductive parameters, in rats. Pregnant rats received three doses of fipronil (0.03; 0.3 or 3 mg/kg) from gestational day 15 to postnatal day 7, by gavage. During the treatment, water and food consumptions as well as the body weight of females exposed were investigated. After weaning, we collected the blood of mothers for biochemical analysis and the organs to verify the weight. The male offspring was evaluated for body weight, anogenital distance and ages of preputial separation and testicular descent. At adult life, weight of reproductive organs, sperm counts, morphology and motility, testicular and epididymal histomorphometry, sertoli cells count, sexual behavior and preference, fertility and patterns of expression of androgen receptor in the testis were evaluated. On female offspring was investigated anogenital distance, puberty onset, estrous cycle, hormonal levels, fertility, sexual behavior and histology of uterus and ovaries. The treatment did not alter the biochemical parameters and the weight of most of analyzed organs of mothers exposed to fipronil, however the weight of pituitary gland presented a reduction in the group exposed to a lower dose. On males, the perinatal exposure to fipronil did not affect the body weight, anogenital distance and puberty onset. Similarly, at adult life, fipronil did not alter the organ... / Doutor

Bacillus sphaericus 2362 is a mosquito pathogenic bacterium. Its greatest industrial potential may be in developing countries where mosquitos are often vectors for diseases. This strain is typical of the species in that it is unable to grow using carbohydrates as a sole source of carbon. The goal of this research was to determine the metabolic deficiency(s) responsible for the inability of this organism to grow on carbohydrates. Compounds that supported light growth of this organism on an agar-solidified, defined medium included acetate, glycerol, and gluconate. Growth in a defined liquid medium with acetate as the source of carbon was much slower than growth in a complex, protein-based broth. B. sphaericus grew poorly in a defined, liquid medium with glycerol or gluconate as the carbon source. Activity of enzymes responsible for the initiation of metabolism of some substrates was not detected in cell extracts. These enzymes were: glucokinase, hexokinase, beta-galactosidase, and amylase. Growth of this bacterium on glycerol as a sole source of carbon implies the presence of the enzymes from the lower half of the Embden-Meyerhof-Parnas (EMP) pathway. Two enzymes of the upper half of the EMF pathway, phosphofructokinase and fructose diphosphate aldolase, were undetected in cell extracts. In addition, glucose dehydrogenase activity was not detected. The inability to form glucose-6-phosphate from glucose prevents the catabolism of this and related substrates via the Entner-Doudoroff (ED), hexose monophosphate (HMP), and EMF pathways. Oxygen uptake studies indicated that B. sphaericus oxidized gluconate slightly but only when the cells were grown in a complex, protein-based medium supplemented with gluconate. Although gluconokinase activity was detected in cell extracts, no activity was detected for the key enzymes of the ED (phosphogluconate dehydratase/KDPG aldolase), or HMP (6-phosphogluconate dehydrogenase) pathway. It is unclear how B. sphaericus grows on a defined medium with gluconate as the sole source of carbon. In addition to enzymatic deficiencies, whole cells were unable to accumulate [¹⁴C]glucose or [¹⁴C]sucrose. / Master of Science