A nutritional method commonly used to deliver feedings to premature infants is the use of a gavage tube. To measure for any undigested breastmilk or formula, a gastric aspirate is checked prior to the next feeding. There is a gap in practice as to what to do if these aspirates signify feeding intolerance. The project question centered on identifying evidence-based guidelines in the literature that would help to define best practices related to feeding intolerance of gavage-fed infants. The Johns Hopkins Nursing Evidence-Based Practice model and the Appraisal of Guidelines Research and Evaluation provided the frameworks for gathering and evaluating evidence as well as the process used in forming the practice guideline. The primary methods employed were a team approach that included a Neonatal Intensive Care Unit (NICU) Project Team and NICU expert opinion along with a literature review conducted by the doctor of nursing practice student. The NICU Project Team collected the NICU experts' input via surveys they developed and distributed as well as e-mails to authors identified from the literature review. The surveys yielded a 76% response rate from the registered nurses and a 59% response rate from the medical providers. All data collected were shared and descriptive statistics were used to evaluate the data. One of the central research findings was that gastric aspirates should no longer be routinely obtained on stable infants and, if used in evaluating feeding intolerance, they must be used in combination with other indicators. An enteral feeding guideline was developed to reflect this finding that can be shared with other NICUs and nurseries in the United States and globally to decrease the morbidity and mortality of neonates.

Disease outbreaks have overwhelmed the aquaculture industry as a whole and have been catastrophic for many single operations. To minimize disease outbreaks, efforts are underway to enhance animal health and disease resistance to pathogens without the use of antibiotics. The overall purpose of this study was to explore a potential prophylactic, a novel antibody diet, for the bacterial pathogen, Aeromonas hydrophila, in tilapia. The tilapia were on an anti-interleukin-10 antibody diet as a neutralization of interleukin-10 (IL-10), an anti-inflammatory cytokine. The anti-inflammatory function of IL-10 has been shown to allow persistence of gastrointestinal pathogens. Tilapia were fed the novel diet and were challenged via bath immersion or oral gavage with A. hydrophila. Four trials of challenge studies were conducted. Clinical signs of the disease and survival were monitored post-challenge of the bacteria. Out of the 4 trials, one bath immersion trial showed significantly lower survival in the group fed the novel antibody diet (p=0.044) compared to the control fed group, after challenged with A. hydrophila. The other trials tested showed no significant differences in survival between diets. Among the survival percentages collected as a whole, it cannot be determined from in vivo results whether this anti-IL-10 diet is effective in preventing mortality from A. hydrophila in tilapia. Therefore, an in vitro study using an enzyme-linked immunosorbent assay (ELISA) was used to determine the neutralization capability of anti-IL-10 on IL-10 using tilapia splenocytes. Interferon-γ, a pro-inflammatory cytokine, was quantified in order to find a trend in expression of IL-10 in vitro in various tilapia cell treatments. The protocol for the ELISA study is under development being that the use of this antibody is novel and has never before been done in fish. / Master of Science in Life Sciences

As alterações respiratórias durante o aumento do volume gástrico, como na dieta por gavagem, acarretam algumas conseqüências ao recém-nascido pré-termo, alterando indicadores da função respiratória, como: aumento da freqüên cia respiratória e cardíaca, fadiga muscu lar, apnéias, cianose, hipoxemia e hiper capnia. Um posicionamento corporal adequado apresenta grande influência sobre a biomecânica da caixa torácica, pois serve como ponto de apoio à mus culatura respiratória, ocasiona maior contratibilidade do músculo, altera a pressão pleural, aumenta o volume de ar pulmonar gerando menor esforço respi ratório e menor desvantagem mecânica ao sistema respiratório. O posicionamen to direcionado aos recém-nascidos pre maturos promove uma reorganização sensorial, sono tranqüilo, menor gasto energético e menor desordem no equilíbrio muscular postural, promovendo menor índice de complicações futuras no desenvolvimento. O presente estudo propôs analisar através dos indicadores da função respiratória a resposta aos posicionamentos em recém-nascidos pré-termo submetidos ao aumento do volume gástrico durante a gavagem. Foi realizado um estudo com delineamento prospectivo, randomizado e tipo cross over. Foram estudados 16 recém-nasci dos prematuros de 31 a 34 semanas de idade gestacional, com peso de nasci mento até 2.500 g. Foram selecionados os recém-nascidos que recebiam a dieta via sonda oro-gástrica com volume total de 150 ml/kg/dia, que se encontravam do sétimo ao décimo dias de vida, sem oxigenoterapia suplementar. Foi utilizado a cada administração da dieta um posicionamento de decúbito diferente, sendo todos com 30° de elevação. Cada recém-nascido recebeu quatro gava gens com intervalo entre elas de 3 ho ras. Os decúbitos foram: lateral direito, lateral esquerdo, pronação e supinação. Os indicadores da função respiratória analisados foram: freqüência respirató ria e cardíaca, saturação de oxigênio, tiragem intercostal, batimento de asa nasal e gemência. Os parâmetros foram coletados de dois em dois minutos a partir de cinco minutos antecedendo a gavagem até o quinto minuto após o término da administração da dieta. Os recém-nascidos acompanhados apre sentaram média de 32 semanas de idade gestacional (desvio padrão, DP, 1,31), peso médio ao nascimento de 1.722 g (DP 276,33). Na comparação entre os decúbitos, a freqüência respiratória média apresentou-se com valores mais altos durante toda a coleta em supina ção e lateral esquerdo (média = 69 e 68, incursões por minuto, ipm) e os valores mais baixos foram encontrados em lateral direito e pronação (61 e 59 ipm). Nos posicionamentos em lateral direito e pronação a freqüência cardíaca média obteve os níveis mais estáveis e mais baixos durante toda a coleta, porém, a supinação apresentou os valores mais altos (média = 142,4, batimentos por minuto, bpm). No decúbito lateral esquerdo e supinação a saturação de oxigênio médio apresentou os menores valores (média geral = 94%) principal mente após a gavagem. Na tiragem intercostal, no batimento de asa nasal e na gemência o lateral direito e pronação apresentaram maior ausência destes sinais, e o lateral esquerdo e supinação apresentaram maior freqüência a eles. Assim, conclui-se que os decúbitos lateral direito e pronação demonstraram maior repercussão positiva nos indica dores da função respiratória, sendo a supinação e lateral esquerdo as posi ções de maiores efeitos negativos nos recém-nascidos submetidos ao aumento do volume gástrico. / The respiratory alterations during the gastric volume\'s increasing, as the gavages, cause some consequences to the premature newborn, modifying the respiratory function indication as: increase of the respiratory and cardiac frequency, muscular fatigue, apneas, cyanosis, hypoxemia and hypercapnia. An appropriate corporal positioning presents great influence on the biomechanics of the rib cage, abutment to the respiratory musculature; it causes greater muscle contraction, modifies the pleural pressure, it increases the pulmonary air volume, causing less respiratory effort and less mechanics\' disadvantages to the respiratory system. The positioning directed to the premature newborns promote a sensorial reorganization, calm sleep, less energy expenses and less clutter in the postural muscular balance, promoting less possibility of future complications in the baby\'s development. The purpose of this study is to analyze through the pointers of the respiratory function the reply of the positioning of in newborn premature submitted to the increase of the gastric volume during the gavages feeding. It was made a study with prospective delineation, randomized and crossover type. It was studied 16 preterm infants of 31 - 34 weeks of gestational age, weighting about 2.500 g. It had been selected newborn that received the diet through the oral-gastric sounding with the volume of 150 ml/kg/day, which was found in the seventh to the tenth days of life, without supplemental oxygen-therapy. It was used different positions to each act of receiving the diet, with 30 degrees elevation. Each newborn used four gavages with spaces of time between them of 3 hours. The positions were: right lateral, left lateral, prone and supine. The analyzed indicators of the respiratory function were: respiratory and cardiac frequency, oxygen saturation, drawing of intercostals, beating of nasal wing and grunting. The parameters were collected from two to two minutes from five minutes preceding the gavage until the fifth minute after the administration diet ending. The newborns studied presented about 32 weeks of gestational age (shunting line standard, DP, 1.31), average weight to the birth of 1.722 g (DP 276,33). Comparing the positions, the average respiratory frequency presented higher values during all the collection in supine and left lateral (average = 69 and 68, breaths per minute, rpm) and the lowest values were found in right lateral and prone (61 and 59 rpm). In the average cardiac frequency, the positions in the right lateral and prone were the ones that had kept the lower levels steadiest during all the collection. However, the supine presented the highest values (average = 142,4, beatings per minute, bpm). In the saturation of medium oxygen, the left lateral and supine presented less values mainly after (average generality = 94%) the gavages. In the drawing of intercostals, in the beating of nasal wing and the grunting, the right lateral and prone presented a greater absence of these signals, and the left lateral and supine presented greater frequency of them. Thus, it is concluded that the right lateral and prone positions showed a greater positive repercussion in the pointers of the respiratory function, being the lateral left and supine the positions of bigger negative effects in the submitted newborn to the increase of the gastric volume.

As alterações respiratórias durante o aumento do volume gástrico, como na dieta por gavagem, acarretam algumas conseqüências ao recém-nascido pré-termo, alterando indicadores da função respiratória, como: aumento da freqüên cia respiratória e cardíaca, fadiga muscu lar, apnéias, cianose, hipoxemia e hiper capnia. Um posicionamento corporal adequado apresenta grande influência sobre a biomecânica da caixa torácica, pois serve como ponto de apoio à mus culatura respiratória, ocasiona maior contratibilidade do músculo, altera a pressão pleural, aumenta o volume de ar pulmonar gerando menor esforço respi ratório e menor desvantagem mecânica ao sistema respiratório. O posicionamen to direcionado aos recém-nascidos pre maturos promove uma reorganização sensorial, sono tranqüilo, menor gasto energético e menor desordem no equilíbrio muscular postural, promovendo menor índice de complicações futuras no desenvolvimento. O presente estudo propôs analisar através dos indicadores da função respiratória a resposta aos posicionamentos em recém-nascidos pré-termo submetidos ao aumento do volume gástrico durante a gavagem. Foi realizado um estudo com delineamento prospectivo, randomizado e tipo cross over. Foram estudados 16 recém-nasci dos prematuros de 31 a 34 semanas de idade gestacional, com peso de nasci mento até 2.500 g. Foram selecionados os recém-nascidos que recebiam a dieta via sonda oro-gástrica com volume total de 150 ml/kg/dia, que se encontravam do sétimo ao décimo dias de vida, sem oxigenoterapia suplementar. Foi utilizado a cada administração da dieta um posicionamento de decúbito diferente, sendo todos com 30° de elevação. Cada recém-nascido recebeu quatro gava gens com intervalo entre elas de 3 ho ras. Os decúbitos foram: lateral direito, lateral esquerdo, pronação e supinação. Os indicadores da função respiratória analisados foram: freqüência respirató ria e cardíaca, saturação de oxigênio, tiragem intercostal, batimento de asa nasal e gemência. Os parâmetros foram coletados de dois em dois minutos a partir de cinco minutos antecedendo a gavagem até o quinto minuto após o término da administração da dieta. Os recém-nascidos acompanhados apre sentaram média de 32 semanas de idade gestacional (desvio padrão, DP, 1,31), peso médio ao nascimento de 1.722 g (DP 276,33). Na comparação entre os decúbitos, a freqüência respiratória média apresentou-se com valores mais altos durante toda a coleta em supina ção e lateral esquerdo (média = 69 e 68, incursões por minuto, ipm) e os valores mais baixos foram encontrados em lateral direito e pronação (61 e 59 ipm). Nos posicionamentos em lateral direito e pronação a freqüência cardíaca média obteve os níveis mais estáveis e mais baixos durante toda a coleta, porém, a supinação apresentou os valores mais altos (média = 142,4, batimentos por minuto, bpm). No decúbito lateral esquerdo e supinação a saturação de oxigênio médio apresentou os menores valores (média geral = 94%) principal mente após a gavagem. Na tiragem intercostal, no batimento de asa nasal e na gemência o lateral direito e pronação apresentaram maior ausência destes sinais, e o lateral esquerdo e supinação apresentaram maior freqüência a eles. Assim, conclui-se que os decúbitos lateral direito e pronação demonstraram maior repercussão positiva nos indica dores da função respiratória, sendo a supinação e lateral esquerdo as posi ções de maiores efeitos negativos nos recém-nascidos submetidos ao aumento do volume gástrico. / The respiratory alterations during the gastric volume\'s increasing, as the gavages, cause some consequences to the premature newborn, modifying the respiratory function indication as: increase of the respiratory and cardiac frequency, muscular fatigue, apneas, cyanosis, hypoxemia and hypercapnia. An appropriate corporal positioning presents great influence on the biomechanics of the rib cage, abutment to the respiratory musculature; it causes greater muscle contraction, modifies the pleural pressure, it increases the pulmonary air volume, causing less respiratory effort and less mechanics\' disadvantages to the respiratory system. The positioning directed to the premature newborns promote a sensorial reorganization, calm sleep, less energy expenses and less clutter in the postural muscular balance, promoting less possibility of future complications in the baby\'s development. The purpose of this study is to analyze through the pointers of the respiratory function the reply of the positioning of in newborn premature submitted to the increase of the gastric volume during the gavages feeding. It was made a study with prospective delineation, randomized and crossover type. It was studied 16 preterm infants of 31 - 34 weeks of gestational age, weighting about 2.500 g. It had been selected newborn that received the diet through the oral-gastric sounding with the volume of 150 ml/kg/day, which was found in the seventh to the tenth days of life, without supplemental oxygen-therapy. It was used different positions to each act of receiving the diet, with 30 degrees elevation. Each newborn used four gavages with spaces of time between them of 3 hours. The positions were: right lateral, left lateral, prone and supine. The analyzed indicators of the respiratory function were: respiratory and cardiac frequency, oxygen saturation, drawing of intercostals, beating of nasal wing and grunting. The parameters were collected from two to two minutes from five minutes preceding the gavage until the fifth minute after the administration diet ending. The newborns studied presented about 32 weeks of gestational age (shunting line standard, DP, 1.31), average weight to the birth of 1.722 g (DP 276,33). Comparing the positions, the average respiratory frequency presented higher values during all the collection in supine and left lateral (average = 69 and 68, breaths per minute, rpm) and the lowest values were found in right lateral and prone (61 and 59 rpm). In the average cardiac frequency, the positions in the right lateral and prone were the ones that had kept the lower levels steadiest during all the collection. However, the supine presented the highest values (average = 142,4, beatings per minute, bpm). In the saturation of medium oxygen, the left lateral and supine presented less values mainly after (average generality = 94%) the gavages. In the drawing of intercostals, in the beating of nasal wing and the grunting, the right lateral and prone presented a greater absence of these signals, and the left lateral and supine presented greater frequency of them. Thus, it is concluded that the right lateral and prone positions showed a greater positive repercussion in the pointers of the respiratory function, being the lateral left and supine the positions of bigger negative effects in the submitted newborn to the increase of the gastric volume.

acase@tulane.edu

BackgroundAtrazine suppression of the LH surge slowly develops over time and peaks after 4 days; sensitivity to atrazine decreases after 8 or 14 days of dosing. Adaptation of the LH response was correlated with increased phase I and phase II liver enzyme activity/expression. MethodsThe effect of atrazine on the LH surge was evaluated in female Sprague-Dawley rats administered 100 mg/kg/day atrazine by gavage for 1, 2, 3, or 4 consecutive days or 6.5, 50, or 100 mg/kg/day atrazine for 4, 8, or 14 days. ResultsNo statistically significant effects of atrazine were seen on peak plasma LH or LH area under the curve (AUC) after one, two, or three doses of 100 mg/kg/day. Four daily doses of 50 or 100 mg/kg atrazine significantly reduced peak LH and LH AUCs, whereas 6.5 mg/kg/day had no effect. After 8 or 14 days of treatment, statistically significantly reduced peak LH and LH AUC were observed in the 100 mg/kg/day dose group, but not in the 6.5 or 50 mg/kg/day dose groups, although significantly reduced LH was observed in one sample 9 hr after lights-on in the 50 mg/kg/day dose group on day 14. The number of days of treatment required to achieve a significant suppression of the LH surge is consistent with the repeat-dose pharmacokinetics of the chlorotriazines. ConclusionThe apparent adaptation to the effect of atrazine on the LH surge after 8 or 14 days may be related to the induction of phase I or, more likely, phase II metabolism observed in this study after 8 days, or to a decreased sensitivity of the hypothalamic-pituitary-adrenal axis or an homeostatic adaption of the effect of atrazine on the LH surge mechanism. Birth Defects Research 110:246-258, 2018. (c) 2017 The Authors. Birth Defects Research Published by Wiley Periodicals, Inc.

Le microbiote intestinal constitue un élément important pour l’hôte, il est impliqué notamment au niveau immunologique ou physiologique. La connaissance de la composition de ce microbiote est la première étape dans la compréhension des phénomènes qui lui sont associés. Les travaux de cette thèse se sont articulés selon plusieurs objectifs. La première étape a été de faire un état des lieux de la composition du microbiote de deux types génétiques parentaux : le canard Pékin (Anas Platyrhynchos), le canard de Barbarie (Cairina Moschata) ainsi que de leur hybride ; le canard mulard. Nous avons ainsi pu observer des compositions différentes selon le type génétique avec tout de même la prédominance dans chacune des trois espèces de deux phyla : les Firmicutes et les Bacteroidetes. Différentes conditions rencontrées dans l’environnement sont à même de créer un déséquilibre dans la composition du microbiote. Une des conditions possibles est un changement dans l’alimentation ainsi la seconde étape a été de voir l’impact du gavage sur les communautés bactériennes composant le microbiote. Celui-ci induit bien des modifications au sein du microbiote en privilégiant certaines classes bactériennes notamment les Bacilli et les Clostridia selon le type génétique. Un effet du gavage a été montré au niveau du microbiote iléal tandis que l’on retrouve un effet du type génétique ainsi que du gavage mais plus faible que dans l’iléon au niveau des caeca. Deux autres travaux ont été réalisés, le premier concernait l’ajout d’un probiotique (Lactobacillus sakei) sur le microbiote des canards mulards durant la phase de gavage. Nous avons montré que lors de cet ajout, nous observions une forte augmentation des lactobacilli au niveau de l’iléon. Enfin suite aux fortes contaminations retrouvées dans le sud-ouest ainsi que ses effets toxiques montrés dans différentes études, le dernier travail effectué a été de voir l’effet du cadmium sur le microbiote des canards Pékin et Barbarie. Nous avons pu ainsi observer les modifications de la communauté microbienne lors de l’ajout de cadmium ainsi l’accumulation de celui-ci dans les reins au cours du gavage. Des effets combinés entre le cadmium et la période de gavage ainsi qu’avec le type génétique ont été mis en lumière. Une tendance à une accumulation différentielle du cadmium a été observé selon le type génétique. / The intestinal microbiota is an important element for the host; it is particularly involved in immunological or physiological level. Knowledge of the composition of the microbiota is the first step in understanding the phenomena associated with it. The work of this PhD was organized according to several objectives. The first step was to realize a molecular inventory of the microbiota composition of the two parental genetic types: the Pekin duck (Anas Platyrhynchos), the Muscovy duck (Cairina moschata) and their hybrid, the mule duck. We have observed different compositions depending on the genetic type although with a predominance of two phyla: the Firmicutes and Bacteroidetes for all ducks species. Different conditions encountered in the environment are likely to create an imbalance in the composition of the microbiota. One is a change in diet; therefore the second step was to see the impact of overfeeding on bacterial communities. Overfeeding causes many changes in the bacterial microbiota and increase two classes: Bacilli and Clostridia according to the genetic type. The effect of overfeeding has been shown principally on the ileal microbiota while genetics and overfeeding both affected weekly cecal microbiota. We then studied the impact of adding a probiotic strain (Lactobacillus sakei) on the microbiota of mule ducks during the overfeeding period. We could see here a significant effect of this addition only in the ileum with a sharp increase in lactobacilli. Finally, due to high levels of contamination found in the southwest of France and its toxic effects on metabolism shown in various studies, the last work was to see the effect of cadmium on the microbiota of Pekin and Muscovy ducks. We observed changes in the microbial community when adding cadmium and see the accumulation of it in the kidneys during overfeeding. Combined effects between cadmium and the feeding period as well as the genetic type were highlighted. Finally

Orientador: Rinaldo Poncio Mendes / Resumo: A paracoccidioidomicose (PCM) é micose granulomatosa sistêmica causada por fungos termodimórficos do gênero Paracoccidioides. O presente estudo teve como objetivos avaliar a atividade da liriodenina, extraída de Annona macroprophyllata Donn. Sm, sobre isolados clínicos e cepas padrão de fungos do gênero Paracoccidioides e outras espécies que causam micoses sistêmicas e, através de um estudo-piloto realizado em modelo murino, avaliar a absorção da liriodenina e possíveis efeitos indesejáveis. A concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM) da liriodenina foram determinadas pelo método de microdiluição. As alterações celulares causadas pela liriodenina em P. brasiliensis / cepa padrão Pb18 foram avaliadas pela microscopia eletrônica de transmissão (MET) e de varredura (MEV). Em estudo-piloto, quatro camundongos isogênicos albinos, da linhagem BALB/c, foram utilizados para avaliar a absorção e os efeitos indesejáveis da liriodenina. Os animais, sem infecção, foram divididos em dois grupos: grupo 1, com dois camundongos que receberam a dose de 0,75 mg.kg-1 e grupo 2, com dois camundongos que receberam a dose de 1,50 mg.kg-1, em única tomada. Após seis e 12 horas após a administração, os animais foram sacrificados e amostras de sangue foram coletadas para a determinação dos níveis séricos. Os intestinos foram coletados para exame histológico. O teste de sensibilidade in vitro revelou que a liriodenina possui atividade sobre parte dos fungos do gênero P... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Paracoccidioidomycosis (PCM) is systemic granulomatous mycosis caused by thermodymorphic fungi of the genus Paracoccidioides. The present study aimed to evaluate the activity of liriodenine, extracted from Annona macroprophyllata Donn. Sm on clinical isolates and standard strains of fungi of the genus Paracoccidioides and other species that cause systemic mycoses and, through a pilot study conducted in a murine model, to assess the absorption of liriodenine and possible undesirable effects. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (CFM) of liriodenine were determined by the microdilution method. Cellular alterations caused by liriodenin in P. brasiliensis / Pb18 standard strain were evaluated by transmission electron microscopy (SEM) and scanning electron microscopy (SEM). In a pilot study, four albino isogenic mice of the BALB / c strain were used to assess the absorption and undesirable effects of liriodenine. The animals, without infection, were divided into two groups: group 1, with two mice receiving the dose of 0.75 mg.kg-1 and group 2, with two mice receiving the dose of 1.50 mg.kg- 1 , in single outlet. After six and 12 hours after administration, the animals were sacrificed and blood samples were collected for the determination of serum levels. The intestines were collected for histological examination. The in vitro sensitivity test revealed that liriodenine has activity on part of fungi of the genus Paracoccidioides, with MIC v... (Complete abstract click electronic access below) / Doutor

Le microbiote intestinal joue un rôle fondamental chez son hôte. Son étude contribue à la compréhension de différents mécanismes physiologiques chez l’hôte. Suite à l’interdiction d’utiliser des antibiotiques comme facteurs de croissance dans l’alimentation animale, l’utilisation de probiotiques s’est largement répandue pour améliorer les performances zootechniques des animaux. De plus, depuis 1999, des études ont été demandée aux pays producteurs de foie gras de rechercher des méthodes alternatives au gavage. Les travaux de cette thèse se sont donc articulés autour de ces deux objectifs. L’effet de la supplémentation en probiotiques (genre Lactobacillus ou mix de souches) chez des canards mulard durant la phase d’élevage et de gavage a donc été étudié. Si aucun effet des probiotiques sur le métabolisme n’a été démontré, l’expression des marqueurs de l’inflammation a diminué lors du gavage. De plus, nous avons confirmé l’importance du genre Lactobacillus, pendant le gavage, groupe bactérien à fort pouvoir amylolytique. Dans un second temps, la composition du microbiote intestinal de l’oie grise landaise gavée ou suite à une stéatose hépatique spontanée a été étudiée et comparée dans le cadre d’un projet d’alternative au gavage. Pour finir, une étude sur le déclenchement d’une stéatose hépatique chez des oies issues du système de reproduction à l’aide d’un aliment complet de croissance a été réalisée. Ces deux études sur les oies nous ont permis d’observer des similitudes dans les phyla majoritaires du microbiote intestinal qui sont les Firmicutes et les Proteobacteria bien que des différences ont été observées à des niveaux taxonomiques plus bas. L’ensemble de ces travaux ont confirmé la corrélation entre diversité du microbiote intestinal et aptitude à déclencher une stéatose hépatique qu’elle soit issue d’un gavage ou spontanée. / The intestinal microbiota plays a fundamental role in its host. Microbiota studies contribute to understand the different physiological mechanisms in the host. Then, following the ban on antibiotics as growth factors in animal feed, the use of probiotics has increased to improve zootechnical performances. Moreover, since 1999, studies have been requested to the European countries producing “foie gras” to develop alternative methods to overfeeding. The work of this phD was organized according to these two objectives. The effect of probiotic supplementation in ducks during breeding and overfeeding periods was studied. If no impact of probiotics on metabolism has been demonstrated, the expression of markers of inflammation decreased during overfeeding. Moreover, we confirmed the importance of the Lactobacillus genus during overfeeding period, bacterial group with strong amylolytic activities. In a second step, the composition of the intestinal microbiota of the graylag goose after overfeeding period or following a spontaneous hepatic steatosis was studied and compared. Finally, a study on the initiation of hepatic steatosis in graylag geese fed with complete pellets was performed. These two geese studies allowed us to observe the similarities in the major phyla of the intestinal microbiota. Major phylum are still Firmicutes and Proteobacteria although differences were observed at lower taxonomic levels. All of these studies confirmed the correlation between intestinal microbiota diversity and the ability to trigger hepatic steatosis, whether with overfeeding or spontaneous.

Submitted by Adriele Dandara Levorato Vinche (adrielelevorato@yahoo.com.br) on 2018-03-13T14:11:39Z No. of bitstreams: 1 Tese de Doutorado Adriele Dandara Levorato Vinche.pdf: 2200417 bytes, checksum: 23e69a47a9fa5126001d659e0803e67a (MD5) / Approved for entry into archive by Luciana Pizzani null (luciana@btu.unesp.br) on 2018-03-13T18:45:09Z (GMT) No. of bitstreams: 1 vinche_adl_dr_bot.pdf: 2200417 bytes, checksum: 23e69a47a9fa5126001d659e0803e67a (MD5) / Made available in DSpace on 2018-03-13T18:45:09Z (GMT). No. of bitstreams: 1 vinche_adl_dr_bot.pdf: 2200417 bytes, checksum: 23e69a47a9fa5126001d659e0803e67a (MD5) Previous issue date: 2018-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A paracoccidioidomicose (PCM) é micose granulomatosa sistêmica causada por fungos termodimórficos do gênero Paracoccidioides. O presente estudo teve como objetivos avaliar a atividade da liriodenina, extraída de Annona macroprophyllata Donn. Sm, sobre isolados clínicos e cepas padrão de fungos do gênero Paracoccidioides e outras espécies que causam micoses sistêmicas e, através de um estudo-piloto realizado em modelo murino, avaliar a absorção da liriodenina e possíveis efeitos indesejáveis. A concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM) da liriodenina foram determinadas pelo método de microdiluição. As alterações celulares causadas pela liriodenina em P. brasiliensis / cepa padrão Pb18 foram avaliadas pela microscopia eletrônica de transmissão (MET) e de varredura (MEV). Em estudo-piloto, quatro camundongos isogênicos albinos, da linhagem BALB/c, foram utilizados para avaliar a absorção e os efeitos indesejáveis da liriodenina. Os animais, sem infecção, foram divididos em dois grupos: grupo 1, com dois camundongos que receberam a dose de 0,75 mg.kg-1 e grupo 2, com dois camundongos que receberam a dose de 1,50 mg.kg-1, em única tomada. Após seis e 12 horas após a administração, os animais foram sacrificados e amostras de sangue foram coletadas para a determinação dos níveis séricos. Os intestinos foram coletados para exame histológico. O teste de sensibilidade in vitro revelou que a liriodenina possui atividade sobre parte dos fungos do gênero Paracoccidioides, com valores de CIM entre 31,2 e 250 μg.mL-1 e sobre a cepa padrão de Histoplasma capsulatum (CIM de 1,95 μg.mL-1). No entanto, menor atividade foi observada em leveduras da maioria das espécies de Candida ensaiadas, com CIM de 125 a 250 μg.mL-1, e maior atividade sobre as cepas padrões de Cryptococcus neoformans e Cryptococcus gattii (CIM de 62,5 μg.mL-1), enquanto a cepa de Aspergillus fumigatus revelou-se resistente à maior concentração de liriodenina. Deve-se ressaltar a atividade da liriodenina sobre Candida krusei, que é intrinsecamente resistente ao fluconazol e, ao contrário, a ausência de atividade sobre Candida tropicalis. A liriodenina revelou ter atividade fungicida sobre todas as cepas padrão e isolados clínicos que foram inibidos in vitro. A MET revelou alterações citoplasmáticas e danos na parede celular de P.brasiliensis. O estudo-piloto revelou que, após 12 horas da administração da liriodenina, os animais que receberam a dose de 1,50 mg.kg-1 apresentavam gases intestinais e distensão abdominal, achados não observados nos que receberam 0,75 mg.kg-1. Os exames histológicos não evidenciaram alterações de intestino com nenhuma das doses empregadas. Níveis séricos de liriodenina foram detectados em ambos os grupos de animais. No entanto, os animais que receberam 0,75 mg.kg-1 de liriodenina apresentaram aumento da concentração com o passar do tempo, enquanto achado oposto foi observado nos que receberam 1,50 mg.kg-1. Os testes de sensibilidade in vitro indicaram que a liriodenina é uma alternativa promissora no tratamento de várias micoses sistêmicas. O estudo-piloto revelou que a liriodenina é absorvida após administração por gavagem, embora a dose de 1,50 mg.kg-1 tenha induzido alterações funcionais, caracterizadas por formação exagerada de gases e conseqüente distensão abdominal, que talvez possam explicar a diminuição de sua absorção. Os resultados do presente estudo estimulam a realização da avaliação da farmacocinética da liriodenina, de sua ação antifúngica em maior número de isolados de espécies de Candida spp, Cryptococcus spp, Histoplasma capsulatum e fungos do gênero Paracoccidioides e de sua eficácia em infecções experimentais em modelo murino. Por se tratar de composto antifúngico não pertencente às classes hoje utilizadas na terapia antifúngica, pode vir a contribuir com o tratamento de pacientes com infecções fúngicas resistentes. / Paracoccidioidomycosis (PCM) is systemic granulomatous mycosis caused by thermodymorphic fungi of the genus Paracoccidioides. The present study aimed to evaluate the activity of liriodenine, extracted from Annona macroprophyllata Donn. Sm on clinical isolates and standard strains of fungi of the genus Paracoccidioides and other species that cause systemic mycoses and, through a pilot study conducted in a murine model, to assess the absorption of liriodenine and possible undesirable effects. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (CFM) of liriodenine were determined by the microdilution method. Cellular alterations caused by liriodenin in P. brasiliensis / Pb18 standard strain were evaluated by transmission electron microscopy (SEM) and scanning electron microscopy (SEM). In a pilot study, four albino isogenic mice of the BALB / c strain were used to assess the absorption and undesirable effects of liriodenine. The animals, without infection, were divided into two groups: group 1, with two mice receiving the dose of 0.75 mg.kg-1 and group 2, with two mice receiving the dose of 1.50 mg.kg- 1 , in single outlet. After six and 12 hours after administration, the animals were sacrificed and blood samples were collected for the determination of serum levels. The intestines were collected for histological examination. The in vitro sensitivity test revealed that liriodenine has activity on part of fungi of the genus Paracoccidioides, with MIC values between 31.2 and 250 μg.mL-1 and on the standard Histoplasma capsulatum strain (MIC of 1.95 μg .mL-1 ). However, lower activity was observed in yeasts of most Candida species tested, with MIC of 125 to 250 μg.mL-1 , and greater activity on the standard strains of Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 μg. mL-1 ), while the strain of Aspergillus fumigatus was resistant to the highest concentration of liriodenine. The activity of liriodenine on Candida krusei, which is intrinsically resistant to fluconazole and, on the contrary, the absence of activity on Candida tropicalis should be emphasized. Liriodenin reveale over all standard strains and clinical isolates that were inhibited in vitro. MET showed cytoplasmic alterations and damage to P. brasiliensis cell wall. The pilot study revealed that animals receiving the 1.50 mg.kg-1 dose had no intestinal gas and abdominal distension 12 hours after administration of liriodenine, which were not observed in those receiving 0.75 mg.kg-1 . Histological examinations showed no intestinal changes at any of the doses used. Serum levels of liriodenine were detected in both groups of animals. However, animals receiving 0.75 mg.kg-1 liriodenine showed increased concentration over time, while the opposite finding was observed in those receiving 1.50 mg.kg-1 . In vitro sensitivity tests indicated that liriodenine is a promising alternative in the treatment of various systemic mycoses. The pilot study revealed that liriodenine is absorbed after administration by gavage, although the 1.50 mg.kg-1 dose induced functional alterations, characterized by exaggerated gas formation and consequent abdominal distension, which may explain the decrease of its absorption. The results of the present study stimulate the evaluation of the pharmacokinetics of liriodenine, its antifungal action in a greater number of isolates of Candida species, Cryptococcus spp, Histoplasma capsulatum and fungi of the genus Paracoccidioides and their efficacy in experimental infections in murine model. Because it is an antifungal compound that does not belong to the classes currently used in antifungal therapy, it may contribute to the treatment of patients with resistant fungal infections.