L'amélioration de la digestion de lactose est une allégation "santé" liée aux ferments viviants du yaourt : Streptococcus thermophilus (S. thermophilus) et Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) validée par l'EFSA en 2010. La physiologie de S. thermophilus et de L. bulgaricus est connue dans le lait et particulièrement le yaourt, alors qu'elle n'a été que peu étudiée dans le tractus digestif (TD). Mon travail de thèse est basé sur l'hypothèse de travail suivante : l'utilisation de modèles animaux gnotobiotiques permet de mieux connaître la physiologie des bactéries lactiques et de proposer des mécanismes d'action de leurs effets "santé". La stratégie a donc été d'obtenir des animaux mono-associés avec chacune des deux bactéries du yaourt ou les deux en même temps. Les principaux résultats obtenus sont : 1/ S. thermophilus colonise le TD en s'adaptant progressivement à l'environnement colique et y induit une glycolyse massive et une production de lactate. La glycolyse est la signature majeure de S. thermophilus dans le TD et le lactate pourrait être est la molécule "signal" qui induit une réponse chez l'hôte par une augmentation des transporteurs de mono-carboxylates (SLC16A1 et SLC5A8) et d'une protéine impliquée dans l'arrêt du cycle cellulaire p27kip1. 2/ L'apport de lactose stimule la colonisation du TD, la glycolyse ainsi que la production de L-lactate par S. thermophilus in vivo. 3/ Contrairement à ce qui est observé pour S. thermophilus, L. bulgaricus ne s'implante pas en absence de lactose. Quand les deux bactéries sont en co-culture, S. thermophilus est toujours avantagé numériquement par rapport à L. bulgaricus aussi bien in vitro qu' in vivo. Au niveau nutritionnel, tous nos résultats sont cohérents avec les allégations "santé" du yaourt avec un effet prébiotique du lactose. L'étude d'animaux gnotobiotiques a permis de proposer des nouvelles voies de régulation du métabolisme des sucres de bactéries lactiques et de nouvelles voies moléculaires (via le lactate) par lesquelles des bactéries lactiques et de nouvelles voies moléculaires (via le lactate) par lesquelles des bactéries lactiques pourraient influencer la physiologie de l'hôte. / *

Quatro experimentos foram conduzidos com o objetivo de avaliar o efeito da forma física da ração pré-inicial no desempenho e na alometria de órgãos do sistema digestório de frangos de corte, provenientes de ovos de pesos semelhantes ou distintos, produzidos por matrizes Ross de idades diferentes. Foram utilizados 2.340 ovos para os experimentos 1 e 2 e 2.160 ovos para os experimentos 3 e 4. A composição percentual dos ovos foi avaliada. Os experimentos 1 e 2, assim como os experimentos 3 e 4, ocorreram simultaneamente. O experimento 1 contou com 1.380 pintos de um dia, machos e fêmeas sexados, provenientes de ovos com peso médio de 57,8 g ou 68,8 g, de matrizes com, respectivamente, 29 e 55 semanas. O experimento 3 contou com 1.380 aves provenientes de matrizes com 29 e 55 semanas de idade, sendo o peso médio dos ovos padronizado em aproximadamente 62 g. O delineamento experimental utilizado foi o inteiramente casualizado, com seis tratamentos e cinco repetições, em esquema fatorial 2x3 (idade da matriz x forma física da ração pré-inicial). Na fase pré-inicial, a forma física da ração foi diferenciada em farelada, triturada e micro-peletizada, com fornecimento fixado em 200 g/ave. Do término da fase pré-inicial até os 42 dias de idade, foi fornecida uma ração comum farelada, para todos os tratamentos. Os parâmetros de desempenho avaliados nos experimentos 1 e 3 foram: ganho de peso, consumo de ração, conversão alimentar e viabilidade criatória. O tempo utilizado para o consumo total da ração pré-inicial foi monitorado. Para a avaliação da alometria de órgãos (intestinos delgado e grosso, fígado, pâncreas, proventrículo, moela e saco vitelino), foram utilizados em cada experimento (2 e 4), 240 pintos de um dia, machos e fêmeas sexados, nos mesmos tratamentos adotados nos experimentos 1 (pesos distintos) e 3 (pesos semelhantes). Foram sacrificadas seis aves provenientes de cada idade de matriz ao alojamento e quatro aves por tratamento nos dias três, cinco, sete, 14 e 21, sendo cada ave considerada uma unidade experimental. A gema de ovos provenientes de matrizes adultas foi maior mesmo quando estes apresentavam pesos semelhantes aos de matrizes jovens. Em ambos os experimentos de desempenho, melhores resultados foram obtidos pelos frangos provenientes de matrizes de 55 semanas de idade, bem como, por aqueles que consumiram rações pré-iniciais processadas. As aves levaram menos tempo para consumir 200 g de rações processadas do que farelada. Ao alojamento, moela, proventrículo, fígado e intestino delgado foram mais pesados para as aves oriundas de matrizes adultas, quando estas apresentavam pesos iniciais superiores aos das aves de matrizes jovens. O desenvolvimento do pâncreas, intestino delgado, fígado e proventrículo na primeira semana de vida foi superior ao crescimento corpóreo das aves, independente da idade da matriz. Conclui-se que o peso do ovo influencia o peso do pinto à eclosão, porém, é a idade da matriz que atua sobre o desempenho de frangos de corte. Além disso, o consumo de dietas processadas na fase pré-inicial favorece o desempenho dos frangos até os 42 dias de idade. / Four trials were conducted to evaluate the physical form of pre-starter diets on performance and allometric growth of gastrointestinal organs of chickens from eggs having the same or different weights, hatched from Ross breeders of different ages. In trails1 and 2, 2,340 eggs were incubated and in trails 3 and 4, 2,160. The eggs percentage composition was evaluated. Trials 1 and 2, as well as trials 3 and 4, were carried out simultaneously. After hatching, 1,380 day-old chicks, males and females, from young breeders (29 weeks) or adult breeders (55 weeks) were used in performance experiments: in experiment 1 the eggs weighted 57,8 g or 68,8 g, respectively, and in experiment 3, egg weight was standardized at 62 g. The performance trials were conducted in a completely randomized design with 6 treatments and 5 replicates, and the treatments consisted of a 2 3 factorial arrangement of breeder age (young and adult) and physical form of the pre-starter feed (mash, crumbled or micro-pellets). This feed was provided at a fixed amount of 200 g per chick and, after that, the birds in all treatments were fed a common starter, grower and finisher diet in mash form, to 42 days of age. Live weight, weight gain, feed intake, feed conversion and viability were evaluated on a pen basis. The time required for total consumption of the pre-starter feeds was monitored. To evaluate the allometric growth of gastrointestinal organs (small and large intestines, liver, pancreas, proventriculus, gizzard and yolk sac), in experiments 2 and 4, 240 one-day-chicks, males and females, were distributed to six pens with the same treatments as experiments 1 and 3, respectively. Six chicks from each breeder age on the first day and four chicks at ages three, five, seven, 14 and 21 days were sacrificed. The experimental unit was each chick. The yolk from adult breeder eggs was heavier, even when the eggs had similar weight of those from young breeders. In both performance trials, better results were obtained for chickens from adult breeders, as well as, for chickens fed processed pre-starter diets. The chickens consumed 200 g of processed diets faster than the mash diet. On the first day, gizzard, proventriculus, liver and small intestine were heavier in chicks from adult breeders than in chicks from young breeders, when the former presented heavier initial weight. Pancreas, small intestine, liver and proventriculus development was superior to body growth, independent of breeder age. It is concluded that egg weight influences the chick weight at hatching, however, breeder age affects the overall performance of the chickens. Furthermore, processed feeds benefit the chicks in the pre-starter phase and this benefit is maintained in the broilers to market weight.

Existe uma série de evidências que os pintos de corte na fase pré-inicial têm seu sistema digestório ainda imaturo, implicando em menor capacidade de aproveitamento dos alimentos. Em função desse fato, ingredientes de maior digestibilidade seriam recomendados para proporcionar um desenvolvimento adequado do organismo. Com isso, o objetivo do presente trabalho foi determinar a energia metabolizável aparente corrigida pelo balanço de nitrogênio (EMAn) de ingredientes como fontes de carboidrato e proteína para frangos de corte na fase pré- inicial, bem como avaliar a utilização destes ingredientes sobre o desenvolvimento inicial de órgãos do trato gastrintestinal, desempenho e características da carcaça das aves. Os ingredientes avaliados foram: milho (MI), amido de milho (AMI), amido de mandioca (AMA), glicose (GLI), lactose (LAC) e sacarose (SAC), como fontes de carboidrato; e farelo de soja (FS), soja integral micronizada (SIM), isolado protéico de soja (IS), ovo em pó (OP), plasma sanguíneo (PS), farelo de glúten de milho (GM) e levedura seca (LS), como fontes de proteína. Para a determinação da EMAn foram desenvolvidos vários ensaios de metabolismo utilizando-se o método de coleta total de excretas. Para a avaliação do desempenho, desenvolvimento do trato gastrintestinal e características da carcaça foram conduzidos diversos experimentos, em gaiolas e em piso de galpão experimental, com a inclusão dos ingredientes nas dietas. Os valores de EMAn para a fase pré- inicial das fontes de carboidrato foram: 3.213; 3.269; 3.690; 3.427; 1.225 e 3.524 kcal/kg para MI, AMI, AMA, GLI, LAC e SAC, respectivamente. Para as fontes protéicas a EMAn foi: 2.085; 4.068; 2.110; 5.095; 3.831; 3.374 e 2.037 kcal/kg para FS, SIM, IS, OP, PS, GM e LS, respectivamente. Para as aves criadas em gaiolas pode-se dizer que o desenvolvimento do trato gastrintestinal, de maneira geral, não foi influenciado pelas diferentes fontes de carboidrato, mas o consumo de ração das aves foi aumentado pelo fornecimento de carboidrato de maior disponibilidade na dieta préinicial, contribuindo para um aumento no peso vivo das aves. As diferentes fontes de proteína afetaram o desenvolvimento dos órgãos, mas esse efeito não foi suficiente para explicar as variações no desempenho das aves. A adição de diferentes fontes de proteína e carboidrato, bem como suas misturas, em dietas pré- iniciais de frangos de corte criados sob piso de galpão experimental, promoveram alterações no desempenho e no desenvolvimento do intestino delgado das aves até o sétimo dia de idade, entretanto as diferenças de desempenho não foram mantidas nas fases subseqüentes, contribuindo para que as características da carcaça e composição da carne de perna não apresentassem diferenças no momento do abate. / There are some evidences in the literature that newly hatched chicks have the digestive system still immature, what result in a small capacity to metabolize feed ingredients. So, more digestible ingredients could be recommended to allow an adequate development of the body. Herewith, the objective of this work was to determine the Ncorrected apparent metabolizable energy (EMAn) of carbohydrate and protein sources for newly hatched chicks, and to evaluate the utilization of these ingredients on the early development of gastrointestinal tract organs, performance and carcass characteristics of birds. The ingredients evaluated were: corn (MI), corn starch (AMI), cassava starch (AMA), glucose (GLI), lactose (LAC) and sucrose (SAC), as carbohydrates sources, and soybean meal (FS), micronized full fat soybean (SIM), isolated soy protein (IS), dried whole eggs (OP), blood plasma (PS), corn gluten meal (GM) and dried sugar cane yeast (LS), as protein sources. Several metabolism trials were conducted to determine the EMAn, using the total excreta collection method. To evaluate the performance, gastrointestinal tract development and carcass characteristics, experiments were conducted, in cages and floor pens, using diets containing those ingredients. The EMAn of the carbohydrate sources determined for chicks in the first week were: 3,213; 3,269; 3,690; 3,427; 1,225 and 3,524 kcal/kg for MI, AMI, AMA, GLI, LAC and SAC, respectively. The EMAn for protein sources were: 2,085; 4,068; 2,110; 5,095; 3,831; 3,374 and 2,037 kcal/kg for FS, SIM, IS, OP, PS, GM and LS, respectively. For the birds reared in cages, the results showed, in general, that the different carbohydrate sources did not affect the gastrointestinal tract development, but the feed consumption of the birds was improved by the more available carbohydrate sources in the newly hatched chicks diets, contributing to a higher liveweight of the birds. Otherwise, the different protein sources affected the gastrointestinal tract development, but these effects did not explain the changes on broiler performance. The addition of different protein and carbohydrate sources, and their mixtures, in the diets of newly hatched chicks reared in the floor pens resulted in differences on performance and on small intestine development of the birds until seven days of age, but these differences were not maintained on the following phases, contributing to a similar carcass characteristics and similar chemical composition of the whole leg meat at slaughter time.

Streptococcus thermophilus est la bactérie lactique la plus utilisée après Lactococcus lactis dans l’industrie laitière pour la fabrication de yaourts et de fromages. Il s’agit du seul streptocoque à avoir le statut de bactérie GRAS (Generally Recognized As Safe). Malgré de récentes études montrant sa capacité à survivre dans le tractus digestif humain et des effets santé intéressants, le statut probiotique de S. thermophilus reste l’objet d’interrogations. Ainsi, les objectifs de cette thèse ont été (i) d’approfondir les connaissances sur la capacité de survie de S. thermophilus en conditions digestives humaines simulées, grâce, en particulier, au système dynamique multi-compartimenté TIM (TNO gastro-intestinal model) et (ii) d’identifier des gènes de S. thermophilus spécifiquement activés dans des conditions complexes comme l’environnement digestif, à l’aide de la technologie R-IVET (Recombinase-based In Vivo Expression Technology) basée sur l’excision d’un gène rapporteur. Le système R-IVET est composé d’un vecteur plasmidique portant la recombinase cre démunie de son promoteur et d’une cassette chromosomique composée d’un gène marqueur entouré de sites loxP reconnus par Cre. Ainsi, dans un premier temps, nous avons implanté la technologie R-IVET chez S. thermophilus LMD-9. Sa fonctionnalité a été testée et validée in vitro et dans le tractus digestif de la souris. Puis, l’étude de la survie de quatre souches de S. thermophilus dans le système TIM a montré que trois d’entre elles étaient plus résistantes que la quatrième, très sensible aux stress gastro-intestinaux. Ces résultats confirment donc que la survie de S. thermophilus dans l’environnement digestif est souche-dépendante. Ils montrent également que la survie de S. thermophilus est influencée par la matrice alimentaire, celle-ci étant plus importante en lait fermenté qu’en lait liquide. Enfin, dans un troisième temps, nous avons construit une première banque génomique R-IVET, en clonant en amont de cre des fragments d’ADN génomiques provenant de LMD-9. Cette banque a été testée uniquement en conditions gastriques simulées dans le TIM. Puis, après avoir optimisé notre outil chez S. thermophilus en améliorant la méthode d’identification des gènes activés, une seconde banque R-IVET a été testée dans l’ensemble du tractus gastro-intestinal (TIM) et en système batch en présence du microbiote intestinal. Ces expériences nous ont permis de mettre en évidence, pour la première fois, des gènes de S. thermophilus spécifiquement activés dans les différents compartiments digestifs de l’homme. Ce travail de thèse contribue ainsi à approfondir les connaissances sur le comportement de cette bactérie dans le tractus gastro-intestinal humain. A moyen terme, ces travaux devraient permettre d’identifier des marqueurs de survie de S. thermophilus et de mieux comprendre son activité métabolique dans l’environnement digestif, facilitant la sélection de souches dans la perspective de développement d’aliments fonctionnels. / Streptococcus thermophilus is the lactic acid bacterium most commonly used after Lactococcus lactis in the dairy industry for the production of yogurt and cheese. It is the only streptococcus strain to have the GRAS status (Generally Recognized As Safe). Despite recent studies showing its ability to survive through the human digestive tract and valuable health effects, the probiotic status of S. thermophilus remains questioned. Thus, the objectives of this pHD work were (i) to increase knowledge on the survival of S. thermophilus in human digestive environment, by using the dynamic multi-compartmental TIM system (TNO gastro-intestinal model) and (ii) to identify the genes from S. thermophilus that are specifically activated in complex digestive environment using the R-IVET technology (Recombinase-based In Vivo Expression Technology). R-IVET is based on the excision of a reporter gene and consists of a plasmid vector carrying the promoterless recombinase cre and a chromosomal cassette composed of a marker gene flanked by loxP recognized by Cre. First, we introduced the R-IVET technology in S. thermophilus LMD-9. Its functionality was tested and validated in vitro and in the mice digestive tract. Then, the survival of four S. thermophilus strains was investigated in the TIM system and we showed that 3 of these strains were more resistant than the other one, very sensitive to gastrointestinal stresses. These results strengthen the idea that the survival of S. thermophilus is strain-dependent. We also highlighted that the survival of S. thermophilus was influenced by the food matrix, being higher in fermented compared to liquid milk. Lastly, we constructed a first genomic R-IVET library, by cloning upstream of cre genomic DNA fragments from LMD-9. This library was tested only in gastric condition (TIM). After optimization of our tool in S. thermophilus (improvement of the method allowing identification of the activated genes), a second R-IVET library was tested throughout the gastrointestinal system (TIM) and in batch system including intestinal microbiota. By identifying bacterial genes specifically activated in human digestive conditions, this work contributes to extend our knowledge on the behavior of S. thermophilus in the human gastrointestinal tract. This could open up opportunities in determining survival markers for S. thermophilus and better describing its metabolic activity in the human gut, then facilitating the selection of strains that can be included in functional foods.

Submitted by (edna.saturno@ufrpe.br) on 2016-11-24T14:28:14Z No. of bitstreams: 1 Franklin Magliano da Cunha.pdf: 1244821 bytes, checksum: e00050f2f8c4d369ea9fc3b76f6cad2e (MD5) / Made available in DSpace on 2016-11-24T14:28:14Z (GMT). No. of bitstreams: 1 Franklin Magliano da Cunha.pdf: 1244821 bytes, checksum: e00050f2f8c4d369ea9fc3b76f6cad2e (MD5) Previous issue date: 2007-02-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / About 10% of the termites’ species have been pointed as agent of some type of damage to the plants. Due to the diversity of food utilization of termites, the characteristics of the digestive tract of the workers and pseudo-workers can be used in systematics and filogenia of these insectes. A control methods of termites is the use of entomopathogenic fungi. Aspects regarding the immunologic response of termites triggered by pathogens invasions, however, remain scarce especially related to the immune reactions deflagrated by the hemocytes. Thus this research focused the following objectives: 1. to identify and to characterize the hemocytes morphology of Nasutitermes coxipoensis (Holmgren) worker´s cast; 2. to generate information about the immunologic reactions of hemocytes when challenged by Metarhizium anisopliae (Metsch.) Sorokin in N. coxipoensis workers and, 3. to describe the histology of workers digestive tractaiming to contribute with support information for systematics of termites. Analysis in light and electronics microscopy of the hemocytes typified the presence of the following cells: Plamatocytes (PL), Granulocytes (GR), Spherulocytes (ES), Prohemocytes (PR), Adipohemocytes (AD) and, Eonocytoids (OE). Among the concentrations from 1x10? to 1x107 conidia/mL of M. anisopliae, the highest concentration 107 conidia/mL was the most efficient with LT50 of 1.62 days. Under this concentration verified greatest frequency of Plamatocytes and Granulocytes in the hemolymph collected during the intervals of from 24 to 72h after application. The hemocitarydynamics during the intervals of 24h, 48h, and 72h after application exhibited significant reduction in Plamatocytes and increasing in Granulocytes. Despite of variation on cell counting was not notified any morphological alterations in the cells. The use of termites spraying as treatment method with 1x107 conidia/mL, in laboratory, triggered the defense mechanisms of N. coxipoensis. The histology of the digestive tract of the workers, in general, exhibited similar characteristics to those observed in other species of termites. Exceptions, however, were identified in the morphology of the epithelial cells regarding the types, number and displaying of these cells in the region of stomodeum as well as the existence of the peritrophic membrane in the region of midgut. Thus these characteristics can be used as criteria for the differentiation among the species of termites of the genus Nasutitermes. / Cerca de 10% das espécies de cupins têm sido apontadas como agente de algum tipo de dano às plantas. Devido ao hábito alimentar variado desses insetos, as características do tubo digestivo dos operários e pseudo-operários são normalmente utilizadas em sistemática e filogenia. Uma alternativa para o controle desses insetos é a utilização de fungos entomopatogênicos. No entanto, os aspectos ligados às reações imunológicas dos insetos promovidas pelas interações com esses patógenos, permanecem ainda bastante escassos, principalmente ao que tange as reações imunes deflagrada pelos hemócitos. Assim a presente pesquisa teve os seguintes objetivos: 1. identificar e caracterizar a morfologia e ultra-estrutura dos hemócitos de operários do cupim Nasutitermes coxipoensis (Holmgren); 2. ampliar as informações sobre as reações imunológicas dos hemócitos quando desafiados imunológicamente por Metarhizium anisopliae (Metsch.) Sorokin. e 3.descrever a histologia do canal alimentar, visando contribuir com informações que possam ser utilizadas na sistemática desse inseto. A análise em microscopia de luz e eletrônica dos hemócitos revelou os seguintes tipos celulares: Plasmatócitos (PL), Granulócitos (GR), Esferulócitos (ES), Prohemócitos (PR), Adipohemócitos (AD) e Oenocitóides (OE). Entre as concentrações de 1x103a 1x107 conídios/mL de M. anisopliae, a concentração mais elevada de 1x107 conídios/mL foi a mais eficiente com TL50 de 1,62 dias. Verificou-se que as células mais freqüentes na hemolinfa foram osPlasmatócitos e os Granulócitos para os tratamentos nos diferentes intervalos de tempo. Analisando a dinâmica hemocitária nos intervalos de 24h, 48h, e 72h, notou-se que os Plasmatócitos reduziram significativamente, enquanto que os Granulócitos aumentaram, após 72h da inoculação. Os Prohemócitos não diferiram estatisticamente nos intervalos de tempos, bem como entre os tratamentos. Durante a contagem diferencial não foram evidenciadas alterações na morfologia dessas células. Assim, a utilização do método de inoculação por pulverização na concentração de 1x107 conídios/mL, em laboratório, mostrou-se eficiente contra os mecanismos de defesa de N. coxipoensis. As características histológicas do canal alimentar dos operários, de modo geral, foram semelhantes as observadas em várias espécies de cupins, porém foram evidenciadas diferenças na morfologia das células epiteliais e tipos de dobras na região do estomodeu, na disposição e númerodos ninhos, bem como a presença da membrana peritrófica na região do mesêntero. Assim essas características podem ser utilizadas como critérios para a diferenciação entre as espécies de cupins do gênero Nasutitermes.

Existe uma série de evidências que os pintos de corte na fase pré-inicial têm seu sistema digestório ainda imaturo, implicando em menor capacidade de aproveitamento dos alimentos. Em função desse fato, ingredientes de maior digestibilidade seriam recomendados para proporcionar um desenvolvimento adequado do organismo. Com isso, o objetivo do presente trabalho foi determinar a energia metabolizável aparente corrigida pelo balanço de nitrogênio (EMAn) de ingredientes como fontes de carboidrato e proteína para frangos de corte na fase pré- inicial, bem como avaliar a utilização destes ingredientes sobre o desenvolvimento inicial de órgãos do trato gastrintestinal, desempenho e características da carcaça das aves. Os ingredientes avaliados foram: milho (MI), amido de milho (AMI), amido de mandioca (AMA), glicose (GLI), lactose (LAC) e sacarose (SAC), como fontes de carboidrato; e farelo de soja (FS), soja integral micronizada (SIM), isolado protéico de soja (IS), ovo em pó (OP), plasma sanguíneo (PS), farelo de glúten de milho (GM) e levedura seca (LS), como fontes de proteína. Para a determinação da EMAn foram desenvolvidos vários ensaios de metabolismo utilizando-se o método de coleta total de excretas. Para a avaliação do desempenho, desenvolvimento do trato gastrintestinal e características da carcaça foram conduzidos diversos experimentos, em gaiolas e em piso de galpão experimental, com a inclusão dos ingredientes nas dietas. Os valores de EMAn para a fase pré- inicial das fontes de carboidrato foram: 3.213; 3.269; 3.690; 3.427; 1.225 e 3.524 kcal/kg para MI, AMI, AMA, GLI, LAC e SAC, respectivamente. Para as fontes protéicas a EMAn foi: 2.085; 4.068; 2.110; 5.095; 3.831; 3.374 e 2.037 kcal/kg para FS, SIM, IS, OP, PS, GM e LS, respectivamente. Para as aves criadas em gaiolas pode-se dizer que o desenvolvimento do trato gastrintestinal, de maneira geral, não foi influenciado pelas diferentes fontes de carboidrato, mas o consumo de ração das aves foi aumentado pelo fornecimento de carboidrato de maior disponibilidade na dieta préinicial, contribuindo para um aumento no peso vivo das aves. As diferentes fontes de proteína afetaram o desenvolvimento dos órgãos, mas esse efeito não foi suficiente para explicar as variações no desempenho das aves. A adição de diferentes fontes de proteína e carboidrato, bem como suas misturas, em dietas pré- iniciais de frangos de corte criados sob piso de galpão experimental, promoveram alterações no desempenho e no desenvolvimento do intestino delgado das aves até o sétimo dia de idade, entretanto as diferenças de desempenho não foram mantidas nas fases subseqüentes, contribuindo para que as características da carcaça e composição da carne de perna não apresentassem diferenças no momento do abate. / There are some evidences in the literature that newly hatched chicks have the digestive system still immature, what result in a small capacity to metabolize feed ingredients. So, more digestible ingredients could be recommended to allow an adequate development of the body. Herewith, the objective of this work was to determine the Ncorrected apparent metabolizable energy (EMAn) of carbohydrate and protein sources for newly hatched chicks, and to evaluate the utilization of these ingredients on the early development of gastrointestinal tract organs, performance and carcass characteristics of birds. The ingredients evaluated were: corn (MI), corn starch (AMI), cassava starch (AMA), glucose (GLI), lactose (LAC) and sucrose (SAC), as carbohydrates sources, and soybean meal (FS), micronized full fat soybean (SIM), isolated soy protein (IS), dried whole eggs (OP), blood plasma (PS), corn gluten meal (GM) and dried sugar cane yeast (LS), as protein sources. Several metabolism trials were conducted to determine the EMAn, using the total excreta collection method. To evaluate the performance, gastrointestinal tract development and carcass characteristics, experiments were conducted, in cages and floor pens, using diets containing those ingredients. The EMAn of the carbohydrate sources determined for chicks in the first week were: 3,213; 3,269; 3,690; 3,427; 1,225 and 3,524 kcal/kg for MI, AMI, AMA, GLI, LAC and SAC, respectively. The EMAn for protein sources were: 2,085; 4,068; 2,110; 5,095; 3,831; 3,374 and 2,037 kcal/kg for FS, SIM, IS, OP, PS, GM and LS, respectively. For the birds reared in cages, the results showed, in general, that the different carbohydrate sources did not affect the gastrointestinal tract development, but the feed consumption of the birds was improved by the more available carbohydrate sources in the newly hatched chicks diets, contributing to a higher liveweight of the birds. Otherwise, the different protein sources affected the gastrointestinal tract development, but these effects did not explain the changes on broiler performance. The addition of different protein and carbohydrate sources, and their mixtures, in the diets of newly hatched chicks reared in the floor pens resulted in differences on performance and on small intestine development of the birds until seven days of age, but these differences were not maintained on the following phases, contributing to a similar carcass characteristics and similar chemical composition of the whole leg meat at slaughter time.

Quatro experimentos foram conduzidos com o objetivo de avaliar o efeito da forma física da ração pré-inicial no desempenho e na alometria de órgãos do sistema digestório de frangos de corte, provenientes de ovos de pesos semelhantes ou distintos, produzidos por matrizes Ross de idades diferentes. Foram utilizados 2.340 ovos para os experimentos 1 e 2 e 2.160 ovos para os experimentos 3 e 4. A composição percentual dos ovos foi avaliada. Os experimentos 1 e 2, assim como os experimentos 3 e 4, ocorreram simultaneamente. O experimento 1 contou com 1.380 pintos de um dia, machos e fêmeas sexados, provenientes de ovos com peso médio de 57,8 g ou 68,8 g, de matrizes com, respectivamente, 29 e 55 semanas. O experimento 3 contou com 1.380 aves provenientes de matrizes com 29 e 55 semanas de idade, sendo o peso médio dos ovos padronizado em aproximadamente 62 g. O delineamento experimental utilizado foi o inteiramente casualizado, com seis tratamentos e cinco repetições, em esquema fatorial 2x3 (idade da matriz x forma física da ração pré-inicial). Na fase pré-inicial, a forma física da ração foi diferenciada em farelada, triturada e micro-peletizada, com fornecimento fixado em 200 g/ave. Do término da fase pré-inicial até os 42 dias de idade, foi fornecida uma ração comum farelada, para todos os tratamentos. Os parâmetros de desempenho avaliados nos experimentos 1 e 3 foram: ganho de peso, consumo de ração, conversão alimentar e viabilidade criatória. O tempo utilizado para o consumo total da ração pré-inicial foi monitorado. Para a avaliação da alometria de órgãos (intestinos delgado e grosso, fígado, pâncreas, proventrículo, moela e saco vitelino), foram utilizados em cada experimento (2 e 4), 240 pintos de um dia, machos e fêmeas sexados, nos mesmos tratamentos adotados nos experimentos 1 (pesos distintos) e 3 (pesos semelhantes). Foram sacrificadas seis aves provenientes de cada idade de matriz ao alojamento e quatro aves por tratamento nos dias três, cinco, sete, 14 e 21, sendo cada ave considerada uma unidade experimental. A gema de ovos provenientes de matrizes adultas foi maior mesmo quando estes apresentavam pesos semelhantes aos de matrizes jovens. Em ambos os experimentos de desempenho, melhores resultados foram obtidos pelos frangos provenientes de matrizes de 55 semanas de idade, bem como, por aqueles que consumiram rações pré-iniciais processadas. As aves levaram menos tempo para consumir 200 g de rações processadas do que farelada. Ao alojamento, moela, proventrículo, fígado e intestino delgado foram mais pesados para as aves oriundas de matrizes adultas, quando estas apresentavam pesos iniciais superiores aos das aves de matrizes jovens. O desenvolvimento do pâncreas, intestino delgado, fígado e proventrículo na primeira semana de vida foi superior ao crescimento corpóreo das aves, independente da idade da matriz. Conclui-se que o peso do ovo influencia o peso do pinto à eclosão, porém, é a idade da matriz que atua sobre o desempenho de frangos de corte. Além disso, o consumo de dietas processadas na fase pré-inicial favorece o desempenho dos frangos até os 42 dias de idade. / Four trials were conducted to evaluate the physical form of pre-starter diets on performance and allometric growth of gastrointestinal organs of chickens from eggs having the same or different weights, hatched from Ross breeders of different ages. In trails1 and 2, 2,340 eggs were incubated and in trails 3 and 4, 2,160. The eggs percentage composition was evaluated. Trials 1 and 2, as well as trials 3 and 4, were carried out simultaneously. After hatching, 1,380 day-old chicks, males and females, from young breeders (29 weeks) or adult breeders (55 weeks) were used in performance experiments: in experiment 1 the eggs weighted 57,8 g or 68,8 g, respectively, and in experiment 3, egg weight was standardized at 62 g. The performance trials were conducted in a completely randomized design with 6 treatments and 5 replicates, and the treatments consisted of a 2 3 factorial arrangement of breeder age (young and adult) and physical form of the pre-starter feed (mash, crumbled or micro-pellets). This feed was provided at a fixed amount of 200 g per chick and, after that, the birds in all treatments were fed a common starter, grower and finisher diet in mash form, to 42 days of age. Live weight, weight gain, feed intake, feed conversion and viability were evaluated on a pen basis. The time required for total consumption of the pre-starter feeds was monitored. To evaluate the allometric growth of gastrointestinal organs (small and large intestines, liver, pancreas, proventriculus, gizzard and yolk sac), in experiments 2 and 4, 240 one-day-chicks, males and females, were distributed to six pens with the same treatments as experiments 1 and 3, respectively. Six chicks from each breeder age on the first day and four chicks at ages three, five, seven, 14 and 21 days were sacrificed. The experimental unit was each chick. The yolk from adult breeder eggs was heavier, even when the eggs had similar weight of those from young breeders. In both performance trials, better results were obtained for chickens from adult breeders, as well as, for chickens fed processed pre-starter diets. The chickens consumed 200 g of processed diets faster than the mash diet. On the first day, gizzard, proventriculus, liver and small intestine were heavier in chicks from adult breeders than in chicks from young breeders, when the former presented heavier initial weight. Pancreas, small intestine, liver and proventriculus development was superior to body growth, independent of breeder age. It is concluded that egg weight influences the chick weight at hatching, however, breeder age affects the overall performance of the chickens. Furthermore, processed feeds benefit the chicks in the pre-starter phase and this benefit is maintained in the broilers to market weight.

This study describes, on a comparative basis, the morphology of mechanical (the linguo-laryngeal apparatus) and sensory (Herbst corpuscles and taste buds) specialisations in the upper digestive tract (bill and oropharynx) of the ostrich and emu, with a view to a better understanding of the functional significance of these structures. The ostrich and emu are commercial entities that constitute important niche industries and are farmed intensively throughout South Africa. A lack of information on the mechanical and sensory specialisations of the upper digestive tract in these two birds hampers a sound understanding of food selection and intake. A total of 48 adult (12-14 months) ostrich heads and 48 adult emu (12-14 months) heads obtained from birds at slaughter at commercial abattoirs and farms, as well as 5 ostrich chick (2-4 weeks) heads and 1 emu chick (8 weeks) head, obtained from previous research projects, were used for this study. Morphological features were described using basic gross anatomical (dissection and stereomicroscopy) and histological techniques (H&E staining), supplemented by differential staining for cartilage and bone, transmission electron microscopy and immunohistochemistry. The findings of the study were compared with the relevant literature and hypotheses for functional significance were formed. The avian glottis channels air from the oropharynx to the trachea and is situated on an elevated structure, the laryngeal mound. It is imperative that the glottis be protected and closed during swallowing, which in mammals is achieved by covering the glottis with the epiglottis, as well as by adduction of the arytenoid cartilages. An epiglottis, however, is reportedly absent in birds. Ratites such as the ostrich and emu possess a very wide glottis in comparison to other birds. The question therefore arises as to how these large birds avoid inhalation of food particles through a wide glottis, with apparently little protection, particularly as their feeding method involves throwing the food over the glottis to land in the proximal esophagus. In the ostrich, when the glottis was closed and the tongue body retracted, the smooth tongue root became highly folded and the rostral portion of the laryngeal mound was encased by the pocket in the base of the ∩-shaped tongue body. In this position the lingual papillae also hooked over the most rostral laryngeal projections. However, in the emu, retraction of the tongue body over the closed glottis resulted in the prominent, triangular tongue root sliding over the rostral portion of the laryngeal mound. In both the ostrich and emu these actions resulted in the rostral portion of the laryngeal mound and weakest point of the adducted glottis being enclosed and stabilised. Only after conducting a comparative study between these two birds using fresh specimens did it become clear how specific morphological peculiarities were perfectly specialised to assist in the closure and protection of the wide glottis. A unique anatomical mechanism in ratites was identified, described and proposed, which may functionally replace an epiglottis; the linguo-laryngeal apparatus. The oropharynx of the ostrich and emu is richly supplied with Herbst corpuscles. This widespread distribution of these mechanoreceptors has not previously been reported in birds. Specific concentrations of Herbst corpuscles within the oropharynx, which differ between the ostrich and emu, assist in the accurate positioning of the tongue and laryngeal mound for cleaning the choana (internal nares). The Herbst corpuscles are strategically located to aid in the handling and transport of food and the median palatine and ventral ridges in the ostrich display a concentration of Herbst corpuscles which denote these structures as sensory organs, namely the palatal and interramal organs. Three specific arrangements of Herbst corpuscles were noted in the oropharynx. The first arrangement consisted of groups of corpuscles located peripherally around a myelinated nerve and was present in the bill tip. The second arrangement, possibly linked to the first, was that of individual or groups of corpuscles without an obvious associated nerve and was present throughout the remaining regions of the oropharynx. The third arrangement was that of corpuscles associated with large, simple branched tubular mucus-secreting glands. The basic structure of Herbst corpuscles in the ostrich and emu, observed by light and transmission electron microscopy, of a capsule (with cellular and acellular lamella), an outer zone (collagen fibrils, fibroblasts and a fluid matrix), an inner core (formed by bilaterally symmetrical specialised Schwann cells) and a receptor axon, is similar to that noted for other avian species. However, unlike in other birds, the capsule of the Herbst corpuscle in the ostrich and emu is formed by myofibroblasts which indicates contractile properties for this component of the corpuscle in ratites. Sensory cilia were noted in the myofibroblasts of the capsule and fibroblasts of the outer zone of the ostrich Herbst corpuscle which may assist in regulating the tension of the capsule. These features have not been reported in other avian species. Although the structure of the palaeognathous palate has been widely studied, relatively little information is available on the morphology of the ratite bill. The kiwi possesses a bill tip organ and the present study confirmed the existence of this somatosensory organ in the ostrich and emu. Examination of the rhamphotheca of these two birds demonstrated numerous specialisations. In the emu, rhamphothecal serrations with intervening keratinised pegs on the rostral mandibular tomia resembled a form of pseudo-teeth. These structures may share a similar embryological origin to teeth; however, they would appear to function by channelling and enhancing vibratory stimuli to Herbst corpuscles in nearby bony pits. In the ostrich, epidermal troughs were present in the regions overlying the bill tip organ and functioned to enhance vibratory stimuli to the underlying Herbst corpuscles. Additionally, in the ostrich only, and not related to the structure or functioning of the bill tip organ, the rostral tomia and maxillary and mandibular nails were composed of typical tubular and inter-tubular horn. This may represent a unique feature in birds. The structure of the mandible and premaxilla was similar to that described previously for these birds. However, the persistence of Meckel’s cartilage through to the adult bird in the ostrich and emu is a novel avian feature not previously reported. The bony bill tips were adorned with numerous sensory (bony) pits which displayed similar distribution patterns in the ostrich and emu and indicated the presence, macroscopically, of a bill tip organ. The total number of pits in the bill tip of the ostrich and emu did not differ significantly, although regional differences did occur. The sub-divisions of the trigeminal nerve (N. opthalmicus R. medialis and N. intramandibularis) innervating the bill tip were well developed in both birds and displayed extensive branching. The emu displayed more myelinated nerve fibres in both nerves than in the ostrich. As myelinated nerve fibres supply Herbst corpuscles, the number of nerve fibres is correlated to the number of corpuscles. No correlation could be made between the number of pits in a particular region and the number of nerve fibres or with the relative percentage of Herbst corpuscles in that region. The bill tip organ in both species was basically similar except for the epidermal specialisations noted above. Two parts of the bill tip organ were recognised; the bony bill tip organ (Herbst corpuscles stacked in bony cavities and pits) and the peripheral bill tip organ (Herbst corpuscles in sheets or chains in the connective tissue between the epithelium and bone). The morphology of the bill tip organ in the ostrich and emu indicates that it is an organ that functions by direct touch. These two ratite species appear to possess the most elaborate bill tip organ of any pecking bird. The existence of a bill tip organ in the ostrich and emu is an enigma and points to the possibility that a bill tip organ is a basal structure in all palaeognathous birds (living and extinct). Furthermore, it is evident by observing the exploratory behaviour of the ostrich and emu, that they use their bill tip organ extensively as a tool for exploring and interpreting their environment as well as for discriminating food. The sense of taste in birds is an important motivator for feeding as well as initial food selection. The existence of this sense in ratites has remained largely speculative. In the present study taste buds were only identified in the emu and were predominantly located in the caudal region of the non-pigmented oropharyngeal roof and sparsely located on the oropharyngeal floor. The taste buds extended the full width of the epithelium in which they were located and were ovoid structures. The taste bud was composed of centrally located, vertically oriented light and dark cells (representing both receptor cells and supporting elements) and peripherally situated follicular cells which were continuous with the surrounding Str. germinativum of the stratified squamous epithelium. Positive IHC labelling for neurofilament demonstrated numerous fine nerve fibres (Neurofibra gustatoria) within the connective tissue immediately surrounding the taste bud. Taste bud morphology in the emu was similar to that described in other birds. However, when sectioned tangentially they were indistinguishable from the surrounding epithelium with H&E staining. By using IHC labelling, concentrations of nerve fibres could be demonstrated beneath apparently nondescript epidermal structures, thus indicating the presence of a taste bud. The distribution of taste buds in the oropharynx could be linked to the particular feeding method of the emu. Based on information from GenBank, it would appear that the relatively few taste buds present in the emu oropharynx would mainly function in distinguishing bitter taste. As bitter-tasting compounds can cause a negative association with a particular food type, it would appear that the sense of taste in the emu would predominantly function for protection and not food selection. This study revealed various unique findings regarding the mechanical and sensory specialisations in the upper digestive tract of the ostrich and emu.  The ostrich and emu possess a combination of structures which functionally replace an epiglottis, namely the linguo-laryngeal apparatus.  Herbst corpuscles are widely distributed in the oropharynx of the ostrich and emu and their distribution is related to the particular feeding habits of these birds.  The capsule of Herbst corpuscles in the ostrich and emu is composed of contractile elements, a feature not reported in other birds.  The ostrich and emu possess a well-developed bill tip organ, which is an unusual feature amongst pecking birds.  Taste buds are present in the emu and no structures resembling taste buds were identified in the ostrich. / Thesis (PhD)--University of Pretoria, 2013. / gm2014 / Anatomy and Physiology / Unrestricted

The University Hospital of Liège has 955 beds in 8 intensive care units, 15 medical wards, 10 surgical wards and 1 paediatric ward. Approximately 36,000 patients are admitted each year, giving a total of 265,000 patient-days hospitalization. Extended-spectrum beta-lactamase-producing Enterobacteriaceae (E-ESBL) constitute, along with methicillin-resistant Staphylococcus aureus (MRSA), the main multi-resistant bacteria recovered in our hospital. The aims of the present study were to: - evaluate the epidemiology of E-ESBL - evaluate the impact of an infection control programme to reduce the spread of E-ESBL in the University Hospital of Liège. In order to do this, several studies were carried out between 2001 and 2007: 1. Determination of the high risk units in the CHU (2001) The high risk units were determined by comparing the incidence rates of each type of unit. Two types of high risk unit were identified in this way: the Intensive Care Units (ICUs) and the Onco-Haematology Unit. 2. Epidemiology of E-ESBL in the Onco-Haematology unit (2002-2003 and 2005-2006) Digestive tract colonization by E-ESBL was found to be relatively high (7.3%) and this explains the high incidence of E-ESBL in Onco-Haematology in comparison with the rest of the hospital. However, the clinical gravity associated with exposure to the risk factor (digestive tract colonization by E-ESBL) was found to be relatively weak. 3. Importance of digestive tract colonization by E-ESBL in General ICUs (2002-2003) Digestive tract colonization by E-ESBL was found to be relatively common (8.8%) and faecal carriage of E-ESBL was found to be a good marker for infection with E-ESBL at another body site. Even though the number of infected patients was found to be low, the risk of infection due to E-ESBL was multiplied by 14.7 in a group of digestive carriers of E-ESBL with regard to a group of non carriers. Our data also showed that Enterobacter aerogenes is the most frequent species producing extended-spectrum beta-lactamase (ESBL) and that TEM-24 is the most prevalent ESBL produced by E-ESBL species in our ICUs. No CTX-M-type genes were identified. With regard to antibiotic susceptibility, meropenem and cefepime appeared to be the most active agents against the majority of isolates. 4. Impact of an infection control programme to reduce the spread of E-ESBL (2006-2007) A surveillance programme was carried out to evaluate the implementation of infection control procedures including surveillance of ESBL-producing strains, utilization of computer alerts for E-ESBL positive patients and the application of contact precautions for colonized or infected patients. Infection control compliance observations were performed by trained referring nurses. During the 2 years of application, one or more E-ESBL were identified in 500 patients. A total of 2268 internal messages regarding the identification of E-ESBL were sent within the hospital, among which 91.84 % were received (at least 1 for every patient). An alert was associated with 406 patients, who were always hospitalized as the identification of the E-ESBL by the laboratory was obtained. A total of 257 registration forms were filled in by the referring nurses, resulting in a survey compliance of 63%. This survey showed that door signs identifying positive patients, hydro-alcoholic solution and gloves were present in 90% of the cases, but that gowns were only present in 59%. The overall incidence of nosocomial acquisition of E-ESBL between 2006 and 2007 was 0.92/1000 patient-days, more or less the same as in 2002. In relation to this research, several questions remain: - Even though the rates of digestive tract colonization with E-ESBL in the 2 types of high risk unit were found to be more or less the same (7.3 and 8.8%), the impact on infections due to E-ESBL was very different. - Are the infections due either to E-ESBL endogenous infections (owing notably to the use of broad spectrum antibiotics) or to secondary infections (resulting from cross-transmission) or to both? The implementation of an infection control programme to limit the spread of E-ESBL has been based on the limitation of the cross-transmission of these micro-organisms. An enhanced barrier precautions policy has been in place in our institution for 2 years, and we have seen no erosion in compliance. We should not however lose sight of the fact that, whatever the institutional policy for the management of multi-resistant bacteria, the correct application of standard precautions for all patients is the first measure to limit the cross-transmission of all micro-organisms.

Shape memory alloys (SMAs) provide exciting opportunities for miniature actuation systems. As SMA actuators are scaled down in size, cooling increases and bandwidth, improves. However, the inclusion of a bias element with which to cycle the SMA actuator becomes difficult at very small scales. One technique used to avoid the necessity of having to include a separate bias element is the use of local annealing to fabricate a monolithic device out of nickel titanium (NiTi). The actuator geometry is machined out of a single piece of non-annealed NiTi. After locally annealing a portion of the complete device, that section exhibits the shape memory effect while the remainder acts as structural support and provides the bias force required for cycling. This work proposes one such locally-annealed monolithic SMA actuator for future incorporation in a device that navigates the digestive tract. After detailing the derivation of lumped electro-mechanical models for the actuator, a description of the prototyping procedure, including fabrication and local annealing of the actuator, is provided. This thesis presents the experimental prototype actuator behaviour and compares it with simulations generated using the developed models.