Orientador: Pedro Luiz Toledo de Arruda Lourenção / Resumo: Introdução: na prática clínica, nem sempre é fácil obter informações adequadas a respeito do padrão evacuatório. Na população pediátrica, os desafios para caracterização do aspecto das fezes são ainda maiores. Desta forma, é de fundamental importância a utilização de uma ferramenta adaptada para a linguagem e entendimento do público infantil. A Escala de Bristol para Consistência de Fezes Modificada para Crianças (“modified Bristol Stool Form Scale for Children – mBSFS-C”) foi recentemente criada e propõe a redução do número de tipos de fezes da Escala de Bristol original, de 7 para 5, e uma adaptação da linguagem utilizada nos descritores. O uso desta escala foi validado nos EUA, mas para que possa ser utilizada em países que possuam outros idiomas, é mandatória a realização de um processo de tradução, adaptação transcultural e validação. Objetivo: realizar o processo de tradução, adaptação transcultural e validação da mBSFS-C para a língua portuguesa do Brasil. Metodologia: a etapa de tradução e adaptação transcultural foi realizada segundo metodologia aceita internacionalmente que propõe a tradução, retro-tradução e pré-teste da versão traduzida da escala, em uma população de 74 crianças para avaliação do entendimento. A etapa de validação foi realizada através de um ensaio, que envolveu 64 crianças e 25 profissionais da saúde, estruturado para investigar a validade e confiabilidade da versão da escala traduzida. Resultados: a tradução e adaptação da mBSFS-C para o portugu... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Functional constipation (FC) is a major gastrointestional (GI) disorder which affects about 14% population worldwide. However, due to efficacy and safety concerns, more than 50% FC patients are not completely satisfied with current conventional therapies, thus alternative therapies are needed. A substantial part of FC patients have symptom of slow colonic motility, while therapy targeting a single pathway cannot benefit all of them. In this thesis, we searched for novel FC therapeutics from two distinct sources, both of which can improve colonic motility significantly: (1) MaZiRenWan (MZRW), an herb formula from Traditional Chinese Medicine (TCM); and (2) Spexin (SPX), a newly identified neuropeptide that is deregulated in FC. On the basis of efficacy validation for MZRW by randomized, placebo-controlled clinical studies, we investigated the bioactive compounds and pharmacological actions of MZRW. Firstly, a machine-learning based method, namely MOST, was developed to relate bioactive compounds with their mechanism-of-action targets. MOST demonstrated good performance in 7-fold cross-validation (over 87% accuracy) and temporal validation (over 76% accuracy). In the case laxative effect, MOST predicted that acetylcholinesterase (ACHE) was the mechanism-of-action target of aloe-emodin; in vivo studies validated this prediction. Secondly, we analyzed the bioactive compounds and mechanism-of-actions of MZRW with combination of UPLC-QTOF-MS/MS, clustering analysis, organ bath, and MOST approaches. 97 compounds were identified in MZRW extract, and 35 of them can be found in plasma and feces samples of rats with oral administration of MZRW. Chemical space analysis suggested that these compounds can be classified into component groups, while the corresponding pharmacology can be studied with representative compounds. Emodin, amygdalin, albiflorin, honokiol, and naringin were shown to induce spontaneous contractions of rat colonic smooth muscle in vitro. Biological targets in ACh-, estrogen-, prostaglandin-, cannabinoid-, and purine signaling pathways are able to explain the prokinetic effects of representative compounds and component groups. Pharmacological actions of MZRW are mixture of five classic paradigms. Thirdly, the latest results of three-armed, randomized and controlled clinical study showed that MZRW demonstrated comparable efficacy with the first line drug Senna, the first line drug for constipation in HK, during treatment period, both were better than placebo; and the efficacy was more sustainable in follow-up period when comparing that of Senna and placebo. These data suggested the unique pharmacological profile of MZRW for FC. With pharmacometabolomic analysis, we found that change of oleamide is negatively correlated (pearson r = -0.59, p<0.001) with improvement of Complete Spontaneous Bowel Movement (CSBM) in MZRW group, but not in Senna or placebo group. Oleamide is up-regulated in FC patients compared with healthy controls, and MZRW can significantly reduce oleamide in FC patients (n=30), healthy human volunteers (n=23), and in normal mice (n=12) serum, ileum, and colon. The regulation of oleamide by MZRW is possibly via augmenting FAAH-mediated degradation. Lastly, we investigated the possibility to use SPX, the newly identified, FC-associated neuropeptide to change GI motility. The deregulation of SPX has been found in several disorders including FC, however, the metabolic instability of SPX prevent it to be directly used in clinical practices. Our investigation through combination of molecular dynamics (MD) simulations and NMR analysis suggested a β-turn-helix-β-turn (βαβ) conformation for human spexin (hSPX) adopts in solution. Consistent with this conformation, cyclic analogues of hSPX with a disulfide bond between residue 1 and 13, LH101 (CWTPQAMLYLKGCQ-NH2), activated both GalR2 (EC50=1.19 μM) and GalR3 (EC50=1.56 μM) with potency comparable to wild type, and that the acetylation at the N-terminal, LH101(Ac) raises the potency EC50=0.38 μM on GalR2 and EC50=0.39 μM on GalR3. The serum half lives of LH101 (t1/2=355.7 min) and LH101(Ac) (t1/2=1973.7 min) were significantly longer than the wild type (t1/2=66.5 min), and LH101(Ac) induces the contractions of mice intestinal segment in vitro and attenuates the oleamide-induced slow GI motility in vivo. Collectively, our studies in MZRW suggested that estrogen and oleamide signaling pathways are potential new targets to develop novel therapeutics for FC, while lead compounds targeting these pathways could be found from MZRW. The final study suggested CSAs have potential to be developed as new FC therapy by targeting the galanin receptor associated pathway.

Probiotics are live microorganisms which when administered in adequate amounts confer one or more health benefits on the host. Different processing conditions, the acidic condition of the stomach and exposure to hydrolytic enzymes affect the viability and efficacy of probiotic organisms. This study investigated the protective effects of two biopolymers poly-gamma-glutamic acid (γ-PGA) and bacterial cellulose (BC) on probiotics during freeze drying and during exposure to simulated intestinal juices and bile salts. The antibacterial property of Bifidobacterium strains was also investigated against four pathogenic bacteria. γ-PGA, a naturally occurring biopolymer was produced by two bacteria (Bacillus subtilis ATCC 15245 and B. licheniformis ATCC 9945a) in GS and E media, γ-PGA yields of about 14.11g/l were achieved in shake flasks and molecular weight of up to 1620 k Da was recorded, γ-PGA production was scaled up in a fermenter with B. subtilis using GS medium. BC, an edible biopolymer was produced by Gluconacetobacter xylinus ATCC 23770 in HS medium and a modified HS (MHS) medium. A yield of about 1.37g/l was recorded and BC production with MHS medium was used for probiotic application. B. longum NCIMB 8809 B. breve NCIMB 8807 and B. animalis NCIMB 702716 showed the best antimicrobial properties against the investigated pathogens. Survival of Bifidobacterium strains was improved when protected with powdered BC (PBC) although γ-PGA offered better protection than PBC. Viability of B. longum NCIMB 8809, B. breve NCIMB 8807 and B. animalis NCIMB 702716 in simulated gastric juice (SGJ) and simulated intestinal juice with bile salts was improved when protected with 5% γ-PGA and 5% γ-PGA+PBC with a reduction of < 1 Log CFU/ml while a reduction of ≤2 Log CFU/ml was recorded in PBC protected cells. Protecting Bifidobacterium strains with γ-PGA, PBC or a novel γ-PGA + PBC combination is a promising method to deliver probiotic bacteria to the target site in order to confer their health benefits on the host.

Orientador: Izabelle Auxiliadora Molina de Almeida Teixeira / Banca: Gustavo Rezende Siqueira / Banca: Antonello Cannas / Banca: Gilberto de Lima Macedo Junior / Banca: Renata Helena Branco Arnandes / Resumo: O objetivo principal foi determinar a contribuição dos diferentes segmentos do trato gastrointestinal (TGI) para a digestão da fibra. Dois experimentos foram conduzidos, os quais foram determinados o tempo médio de retenção (TMR) de partículas e líquido, conteúdo e pesos dos tecidos nos diferentes segmentos pelo método do abate. O primeiro estudo avaliou TMR de partículas e líquido de cabritos Saanen em crescimento de diferentes sexos e submetidos a diferentes níveis de restrição alimentar. Foram utilizados 54 cabritos da raça Saanen (18 fêmeas, 18 machos castrados e 18 machos inteiros) em um arranjo fatorial 3 × 3 que compreendeu os três sexos e três níveis de restrição alimentar (ad libitum / sem restrição, restrição moderada e severa). Contrastes polinomiais foram utilizados para determinar o efeito linear e quadrático da restrição alimentar, enquanto o efeito do sexo foi comparado pelo teste de Tukey. Os efeitos do sexo e interação entre sexo e restrição alimentar não foram significativos na maioria das variáveis avaliadas. A restrição alimentar aumentou o TMR de partículas e fluído. O TMR foi um recurso importante para aumentar a oferta de nutrientes, quando os animais foram submetidos à restrição alimentar, como indicado pelo aumento da digestibilidade total. O segundo estudo avaliou aspectos fisiológicos da digestão da fibra, retenção de partículas e líquido, conteúdo e peso dos tecidos em todo o TGI de cabritos da raça Saanen. O total de 58 cabritos da raça Saanen foram aleatoriamente dispostos em um arranjo fatorial 3 × 3, composto por três sexos (fêmeas, machos castrados, machos inteiros) e três pesos de abate (aproximadamente 16, 23, e 30 kg de peso no abate). Os efeitos do tratamento foram avaliados em um esquema de parcelas subdivididas, com o sexo como efeito principal e peso de abate como sub parcela. Contrastes polinomiais foram utilizados para determinar os efeitos ... / Abstract: The main objective was to assess the contribution of different segments of the gastrointestinal tract (GIT) to the fiber digestion. Two experiments were conducted, which mean retention time (MRT) of particulate and liquid matter, pool size, and tissues weight in different segments were determined by slaughter technique. The study one evaluated MRT of particulate and liquid matter in growing Saanen goats of different sexes and subjected to different levels of feed restriction. Fifty-four Saanen goats (18 each of females, castrated males and intact males) were used in a 3×3 factorial arrangement comprising the three sexes and three levels of feed restriction (unrestricted/ad libitum, moderate and severe restriction). Polynomial contrasts were used to determine linear and quadratic effect of feed restriction, while the effect of sex was compared by Tukey test. The effects of sex and the interaction between sex and feed restriction were not significant on most of variables evaluated. In conclusion, the level of feed restriction increased the MRT of particulate and liquid matter. The MRT was an important mechanism to increase nutrient supply when animals were subjected to feed restriction, as indicated by increased total tract digestibility. The second study examined physiological aspects in the digestion of fiber, particulate and liquid matter residence, pool size, and tissues weight in the whole tract of growing Saanen goats. A total of 58 Saanen goats were disposed in a 3×3 factorial arrangement consisting of three sexes (female, castrated males, intact males) and three slaughter weights (target BW 16, 23, and 30 kg at slaughter). Treatment effects were evaluated in a split-plot design, with sex as the main plot and slaughter weight as the subplot. Polynomial contrasts were used to determine linear and quadratic effects of slaughter weight. Thus increased BW at slaughter resulted in greater MRT of particulate and liquid matter in ... / Doutor

Orientador: José Ricardo de Arruda Miranda / Banca: Cristina Helena dos Reis Serra / Banca: Oswaldo Baffa Filho / Banca: Raul César Evangelista / Banca: Eryvaldo Sócrates Tabosa do Egyto / Resumo: A administração oral de drogas é uma prática comum na terapia e as formas farmacêuticas sólidas são amplamente utilizadas. A variação no perfil de absorção ao longo do trato gastrintestinal (TGI) humano e a possibilidade de liberar drogas em diferentes regiões são os maiores desafios para o desenvolvimento de novos produtos. Desse modo, avaliar formas farmacêuticas sólidas in vivo fornece um entendimento mais profundo quando um efeito sistêmico ou local é desejado. Geralmente, estes estudos são realizados por meio da cintilografia e técnicas biomagnéticas. A Biosusceptometria de Corrente Alternada (BAC) é uma técnica que merece destaque por suas características, acurácia dos resultados obtidos e versatilidade. A BAC propiciou imagens do processo de desintegração de comprimidos tanto in vitro quanto no estômago humano, introduzindo outra perspectiva na análise desse processo. Os resultados também foram correlacionados com sucesso com aqueles obtidos por metodologias específicas, garantindo uma análise mais acurada dos parâmetros físicos envolvidos com a desintegração de comprimidos. A utilização da BAC permitiu avaliar a motilidade gastrintestinal e o processo de desintegração de cápsulas de hidroxipropilmetilcelulose (HPMC) revestidas no cólon humano. Além disso, também foi possível investigar a influência do estado prandial no esvaziamento gástrico e no trânsito gastrintestinal de um sistema multiparticulado magnético. Todos esses trabalhos fortaleceram a BAC como um método alternativo na pesquisa farmacêutica demonstrando seu potencial para avaliar diferentes processos, apesar das suas limitações. Sintetizando, a BAC é uma ferramenta valiosa, com a vantagem de ser livre de radiação e inócua aos voluntários, e vasta aplicabilidade na pesquisa farmacêutica, farmacológica e fisiológica. / Abstract: Oral administration is widely accepted route for drug delivery and solid dosage forms are commonly administered. The variation of absorption profiles along the human gastrointestinal tract (GIT) and the ability to target drugs by adequate dosage forms to distinct sites is the challenge in the pharmaceutical development of solid dosage forms. An understanding of the factors involved in drug absorption and how the gastrointestinal variables can interfere with this process is important to develop more reliable drug delivery systems. The performance of pharmaceutical dosage forms must be fully investigated in vivo to provide more reliable information when a local or systemic effect is desirable. Generally, in vivo investigation on the behavior of dosage forms has been made by using gamma‐scintigraphy and biomagnetic techniques. AC Biosusceptometry (ACB) deserves consideration due to its features, accuracy and versatility. By using ACB technique, it was possible to monitor the disintegration process through acquisition of magnetic images in vitro and in human stomach. The results also were successfully correlated with those obtained with standard methods which provided a more reliable analysis on the physical parameters involved in the disintegration process of tablets. ACB allowed evaluating the gastrointestinal motility and the disintegration of hydroxipropylmethylcellulose (HPMC) coated capsules in human colon. Moreover, it was possible to investigate the gastric emptying and gastrointestinal transit of a magnetic multiparticulate system under influence of prandial state. All these studies have contributed to establish the ACB as an alternative method for pharmaceutical research and, despite some limitations, it was feasible to evaluate different pharmaceutical processes. In summary, ACB is a radiation free and non‐invasive technique with wide applicability in pharmaceutical, physiological and pharmacological researches. / Doutor

Made available in DSpace on 2016-08-15T20:27:55Z (GMT). No. of bitstreams: 1 AnaCDSB_TESE.pdf: 11862899 bytes, checksum: db329299185e2c022af1f916b746aae8 (MD5) Previous issue date: 2014-11-14 / The increase in anthelmintic resistance in gastrointestinal nematodes on small ruminants became a worldwide issue on the development of livestock. In order to assess the resistance scenario in that context in the Rio Grande do Norte region, in vivo and in vitro resistance tests were carried out in 30 rural properties using chemicals in the avermectin, salicylanides, benzimidazoles and imidatiazols groups. For the in vivo test, group I was used as control and four other experimental groups received oral medication, being group II given albendazole (10 mg/kg), group III levamisole (7.5 mg/kg), group IV closantel (5.0 mg/kg) and group V ivermectin (0.2 mg/kg). The nematode infection was determined using fecal egg count (FEC) essays and the efficiency of each treatment was analyzed by the fecal egg count reduction test (FECRT). For the in vivo e in vitro tests, using tiabendazole, the necessary concentration to inhibit 50% of egg hatching (EC50) and assays with a 50% inhibition of larval development (LD50) were assessed, however 50% of larval development and migration inhibition assays (LD50) essays were carried out using ivermectin. For the molecular analysis Haemonchus contortus/property were isolated for the calculation of the frequency of resistant and sensitive alleles for single nucleotide polymorphisms (SNPs): F220Y, F167Y and E198A. The results obtained from in vivo tests showed that resistance was detected for all of the drugs, with leavamisole and closantel appearing as the most effective treatments with FECRT results of 84.3% and 74.6%, respectively. In the in vitro assays, tiabendazole had a result in the hatching test of EC50 0.598 µg/ml (95% IC: 0.323-0.818), while exhibiting in the larval development test a LD50 and LD99 0.518 µg/ml(95% IC: 0.432-0,887) and 4.359 µg/ml (95% IC: 3.047-8.178), respectively. For ivermectin in the larval development test, DL50 0.637 µg/ml(95% IC: 0.382 0.903 µg/ml) e DL99 19.56 µg/ml(95% IC: 11.44 46.55 µg/ml) was observed, while in the migration essay the LD50 value was 16.337 µM (95% IC: 12.7-21.3 µM). In the molecular analysis a higher frequency of the resistant allele for SNP F200Y was observed, presenting higher percentage values of 85.81% and 73.11%, while the F167Y resistant alleles had lower values ranging from 2.93% to 43.73%. In the E198A SNP, only sensitive alleles were detected. Assessing the scenario of anthelmintic resistance in the region, one notices that it s established for the main existing drugs on the market / O aumento na resistência anti-helmíntica em nematóides gastrintestinais de pequenos ruminantes tornou-se um problema global para o desenvolvimento da pecuária. Nesse contexto, a fim de avaliar o cenário da resistência na região do Rio Grande do Norte foram realizados em 30 propriedades rurais testes de resistência in vivo e in vitro utilizando produtos químicos dos grupos das avermectina, salicilanilidas, benzimidazóis e imidatiazóis. Para o teste in vivo utilizou-se o grupo I controle quatro grupos experimentais, sendo grupo II albendazol (10 mg/kg), grupo III levamisol (7,5 mg/kg), grupo IV closantel (5,0 mg/kg) e grupo V ivermectina (0,2 mg/kg). A infecção por nematóides foi determinada utilizando ensaios de ovos por grama de fezes (OPG) e a eficiência de cada tratamento pelo teste de redução da contagem de ovos nas fezes (FECRT). Para os teste in vitro, foi utilizando tiabendazol, com concentrações necessária para inibir 50% de eclosão dos ovos (EC50) e ensaios com inibição de 50% do desenvolvimento larval, porém com a ivermectina foram realizados ensaios de inibição de 50% do desenvolvimento e migração larval (DL50). Para análise molecular foram isolados Haemonchus contortus em cada propriedade avaliada para cálculo da frequência de alelos resistentes e sensíveis para os polimorfismos de nucleotídeo único (SNPs): F200Y, F167Y e E198A. Os resultados obtidos com os testes in vivo e in vitro mostraram que a resistência foi detectado em todas as drogas testadas, com o levamisol e closantel apresentando-se como os tratamentos mais eficientes, com resultados FECRT de 84,3% e 74,6%, respectivamente. Nas análises in vitro o resultado do tiabendazol no teste de eclosão foi EC50 0,598 µg/ml (IC95%: 0,323-0,818), enquanto que no desenvolvimento larval uma DL50 e DL99 de 0,518 µg/ml(IC95%: 0,432-0,887) e 4,359 µg/ml(IC95%: 3,047-8,178), respectivamente. Para ivermectina no teste de desenvolvimento larval foi observado DL50 0,637 µg/ml(CI95%: 0,382 0,903 µg/ml) e DL99 19,56 µg/ml(CI95%: 11,44 46,55 µg/ml), enquanto que no de migração o valor da DL50 foi 16,337 µM (CI95%: 12,7-21,3 µM). Na análise molecular observou-se uma frequência maior do alelo resistente para o SNP F200Y apresentando porcentagens mais altas com valores de 85,81% e 73,11%, enquanto que o F167Y apresentou valores menores de alelos resistentes variando entre 43,73% e 2,93%. No SNP E198A foram detectados alelos apenas sensíveis. Avaliando o cenário da resistência anti-helmíntica na região, percebe-se que está estabelecida para as principais drogas existentes no mercado

Ecotoxicity testing of chemicals for environmental risk assessment is an area where a high number of vertebrates are used across a variety of industrial sectors. The application of the 3Rs in toxicity testing using fish address both the ethical and societal concerns around this issue in addition to the increasing legislative requests for the incorporation of animal alternatives. This thesis aims to highlight the potential of 3D cell culture models to "bridge the gap" between in vitro and in vivo screening procedures for testing of chemicals with the potential to persist or bioaccumulate thereby improving the predictive power of screening procedures. This thesis examines two alternative methods for their potential use as an intestinal based toxicokinetic tool for environmental risk assessment, utilising an in vitro fish cell line replacement tool (RTgutGC). In addition, for the first time a new intestinal primary cell culture based model was developed to address both intestine region specific response (pyloric, anterior, mid and posterior) and size related adaptability to toxins. Paramagnetic oximetry was used to measure oxygen content within 3D structures (spheroids) in order to better understand the microenvironment of these culture models. Using histology, immunohistochemistry, transepithelial electrical resistance (TEER), transmission electron microscopy (TEM), metabolic, fluorescence and gene expression assays, the comparability of this system to native intestinal response was established. Following exposure to carefully chosen environmental contaminants (Benzo[a]pyrene and Copper), the RTgutGC cell line demonstrated comparable responses to existing literature in terms of uptake, metabolism, DNA damage and the presence an equivalent saturable level. Primary enterocytes cultured on transwell inserts remained viable for upto six weeks, with permeability and metabolic activity comparable to native tissue (both in vitro and ex vivo). Taken in combination, these features of enterocytes represent a profile more closely representative of the intestine then the widely used "gut sac" method. With the potential advantages of incorporating complexity at differing levels (connective tissue layer, intestinal bacteria biome), the intestinal models described offer the potential to screen highly persistent toxins which may require prolonged incubation, in addition to the exploration of complex experimental designs which minimise animal usage (uptake, depuration, uptake). As a consequence, the models developed within this thesis significantly enrich the emerging fish based in vitro testing strategies.

Orientador: José Antônio Marques / Banca: Angélica Trazzi Bento de Moraes / Banca: Paola Castro Moraes / Banca: Júlio Carlos Canola / Banca: Elzylene Léga / Resumo: A detomidina é um dos fármacos mais utilizados para a sedação de muares, porém não há dose estabelecida para esta espécie sendo indicado utilizar uma dose 50% maior que a indicada para equinos. Assim, o objetivo deste estudo foi avaliar a eficácia e os efeitos cardiovasculares da detomidina em muares comparando duas doses diferentes, a utilizada em equinos ( G1 - 0,02 mg/kg) e a dose 50% maior (G2 - 0,03 mg/kg). Para isso, foram utiizados 6 muares (2 machos e 4 fêmeas), adultos (339±27 kg), os animais receberam as duas doses com espaçamento mínimo de 7 dias, formando assim dois grupos experimentais com 6 animais cada. Antes da aplicação (M0) da detomidina pela via intravenosa e 5, 15, 30, 45 e 60 minutos após a aplicação (M5; M15; M30; M45; M60, respectivamente) foram avaliados frequência cardíaca, frequência respiratória, motilidade intestinal, altura da cabeça, além dos parâmetros ecocardiográficos: diâmetro interno do ventrículo esquerdo ao final da sístole e da diástole, fração de encurtamento, fração de ejeção, volume do ventrículo esquerdo ao final da sístole e da diástole, volume sistólico e débito cardíaco. A altura da cabeça do G2 permaneceu significativamente menor até o M60 quando comparado ao M0 do mesmo grupo, já do G1 ficou significativamente menor até o M45 quando comparado ao basal. O débito cardíaco do G1 foi significativamente menor nos M5, M15, M30 quando comparado ao M0 doG1, já do G2 permaneceu significativamente menor, quando compardo ao M0 do G2, do M5 ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Detomidine is the drugs most used for sedation of mules, but there is no established dose for this species being indicated to use a dose 50% greater than indicated for horses. Thus, the objective of this study was to evaluate the efficacy and cardiovascular effects of detomidine in mules comparing two different doses, the used in horses (G1 - 0.02 mg / kg) and the 50% higher dose (G2 - 0.03 mg / kg). For this, 6 muares (2 males and 4 females), adults (339 ± 27 kg) were used, the animals received the two doses with a minimum spacing of 7 days, thus forming two experimental groups with 6 animals each. Before the application (M0) of detomidine intravenously and 5, 15, 30, 45 and 60 minutes after the application (M5, M15, M30, M45, M60, M60, respectively) were evaluated heart rate, respiratory rate, intestinal motility, height of the head, in addition to the echocardiographic parameters: internal left ventricular diameter at the end of systole and diastole, fraction of shortening, ejection fraction, left ventricular volume at the end of systole and diastole, systolic volume and cardiac output. The head height of G2 remained significantly lower until M60, whereas G1 was significantly lower up to M45, cardiac output of G1 was significantly lower in M5, M15, M30, whereas G2 remained smaller than M5 at M45. Concluding that mules receiving a 50% higher dose of detomidine remained sedated for a longer period, but the cardiovascular effects were also longer. / Doutor

No Rio Grande do Sul, um grupo clonal de Salmonella Enteritidis vem sendo identificada como o principal microrganismo causador de Doenças Transmitidas por Alimentos (DTA) nos últimos anos. Em vista disso, o objetivo deste trabalho foi avaliar a sobrevivência em fluido gástrico simulado (FGS) e a capacidade de invasão intestinal de Salmonella Enteritidis (SE86) em comparação à Salmonella Typhimurium (ST99) em dois modelos animais. Os microrganismos foram cultivados em meio de cultura e meio de cultura suplementado com glicose, objetivando a adaptação ácida. Aproximadamente 9logUFC dos microrganismos SE86 e ST99, ácido-adaptados ou não adaptados, foram expostos ao FGS, com pH 1,5, e também inoculados por via oral em ratos Wistar adultos machos. Um inóculo de 2logUFC foi utilizado nos experimentos com camundongos “germ-free”, de 21 dias, mantidos em condições assépticas. As fezes e porções do trato gastrintestinal foram analisadas microbiologicamente, sendo que os ratos foram também investigados quanto a lesões morfológicas intestinais. Os camundongos foram analisados por histopatologia e submetidos à curva de mortalidade. Os resultados indicaram que SE86 ácido-adaptadas apresentaram uma sobrevivência significativamente maior (p <0,05) que às SE86 e ST99 não adaptadas ao ácido e também às ST99 ácidoadaptadas no FGS. Os experimentos “in vivo” demonstraram que as bactérias inoculadas foram capazes de provocar lesões morfológicas intestinais e foram recuperadas das fezes, do jejuno e da junção íleo-cecal dos ratos, sendo que contagens mais altas de SE86 e ST99 ácido-adaptadas foram encontradas nas fezes. SE86 ácido-adaptada demonstrou maior contagem no ileo-ceco do que as outras linhagens, sugerindo que a adaptação ácida influenciou na virulência deste microrganismo. Nos animais “germ-free” houve rápida multiplicação de todos os microrganismos inoculados, contudo a mortalidade ocorreu de forma mais rápida devido à infecção por SE86 ácido-adaptada. A análise histopatológica revelou maior severidade da infecção provocada pela SE86, o que foi confirmado pela mortalidade dos animais a partir do quarto dia de infecção, ao contrário da ST99 que não provocou a morte dos animais por até 12 dias. A maior sobrevivência e virulência apresentadas pela SE86 podem estar relacionadas com o freqüente envolvimento desta cepa com salmoneloses alimentares na última década no Rio Grande do Sul. / In Rio Grande do Sul (RS) State, Southern Brazil, a clonal group of Salmonella Enteritidis has been identified as the main cause of foodborne diseases, in the last years. Given that, the objective of this study was to evaluate the survival in simulated gastric fluid (FGS) and the intestinal invasion ability of Salmonella Enteritidis (SE86) and Salmonella Typhimurium (ST99) submitted or not to acid adaptation. The microorganisms were grown in culture media and culture media supplemented with glucose, aiming to promote acid adaptation. After that, approximately 9log of SE86 (Isolated from a foodborne outbreak) and ST99 (not involved with foodborne outbreaks), acid-adapted or not adapted, were exposed to FGS with pH 1.5, and inoculated in adult male Wistar rats. Germ-free mice were also inoculated but with approximately 2 log, and the animals were observed during 21 days, at aseptic conditions. Animal feces and portions of the gastrointestinal tract were examined by microbiological analysis, and the rats were also investigated for intestinal morphological damage. The mice were analyzed by histopathology and submitted to the mortality curve. The results indicated that acid-adapted SE86 had a significant higher survival rate (p<0.05) than not adapted SE86, not adapted ST99, and also than acid-adapted ST99, in the FGS. The “in vivo" experiments demonstrated that the strains were capable of causing intestinal morphological damage and were recovered from feces, jejunum and ileum-cecal junction of rats, with higher counts being demonstrated for acid-adapted SE86 and acid-adapted ST99. Acid-adapted SE86 showed higher counts in the ileum-cecal junction than the other strains, suggesting that acid adaptation influenced the virulence of this microorganism. After inoculation, all strains were able to rapidly multiply in germ-free animals, but mortality caused by acid-adapted SE86 was more intense. Histopathological analyses revealed greater infection’s severity caused by SE86, and it was confirmed by the death of animals starting at the fourth day of infection, unlike the ST99 which did not cause the death of animals until twelve days of inoculation. Based on these results, the higher survival rates in FGS and the higher virulence presented by the SE86 can be related to the frequent involvement of this strain with foodborne salmonellosis occurred in the last decade in RS.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2016-06-24T09:28:21Z No. of bitstreams: 1 texto completo.pdf: 219807 bytes, checksum: 21f8f8ad7c620819dca3b943cc35dbc0 (MD5) / Made available in DSpace on 2016-06-24T09:28:21Z (GMT). No. of bitstreams: 1 texto completo.pdf: 219807 bytes, checksum: 21f8f8ad7c620819dca3b943cc35dbc0 (MD5) Previous issue date: 1999-02-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Foi conduzido um experimento no setor de caprinocultura do Departamento de Zootecnia da Universidade Federal de Viçosa, com o objetivo de avaliar o efeito do colostro de vaca fermentado com ou sem óleo de soja e com ou sem aplicação de zeranol sobre o desenvolvimento de cabritos pré- ruminantes. O experimento foi constituído de duas partes: na primeira, foram utilizados 24 animais machos da raça pardo alpina dispostos em um delineamento inteiramente casualizado em esquema fatorial 3 x 2, sendo três fontes (fonte 1: leite de cabra; fonte 2: colostro fermentado de vaca; e fonte 3: colostro fermentado de vaca + óleo de soja) e duas combinações com zeranol (com e sem), constituindo seis tratamentos com quatro repetições. Foram avaliados o ganho de peso dos animais, o consumo de concentrado, o consumo de matéria seca e a conversão alimentar. Para ganho de peso, observaram-se diferenças (P<0,05) para peso aos 29, 36 e 50 dias de idade, no entanto, não houve diferenças para ganho médio diário (P>0,05). Também não foram observadas diferenças (P>0,05) para consumo de concentrado, consumo de matéria seca e conversão alimentar. Na segunda parte do experimento, foram abatidos três animais de cada tratamento, com o objetivo de se avaliar o efeito das dietas sobre o desenvolvimento dos órgãos, o comprimento dos intestinos, o peso de corpo vazio e a composição química da carcaça dos animais. Para peso de corpo vazio e peso metabólico, houve efeito significativo (P<0,05) para as fontes utilizadas. Não foram observadas diferenças significativas entre as fontes ou o uso do zeranol. O omaso foi influenciado pelo zeranol, observando-se interação significativa para peso de mesentério. Os pesos dos rins, do pulmão e baço foram influenciados pelo tipo de dieta. Apenas o intestino grosso foi influenciado pela dieta. Concluiu-se que o colostro fermentado de vaca e o uso do óleo de soja apresentam-se como alternativa viável na alimentação de cabritos pré-ruminantes, porém o uso do zeranol não mostrou resultados que justifiquem a sua utilização para estes animais. / An experiment was carried out to evaluate the effect of the fermented cow colostrum with or without soybean oil and with or without zeranol application on the pre-ruminant goats development. The experiment was constituted of two parts; in the first 24 Alpine brown males were allotted to a completely randomized design in a factorial 3 x 2 were used, being three sources (source 1: goat milk; source 2: cow fermented colostrum; source 3: cow fermented colostrum + soybean oil) and two arraignments with zeranol (with and without), constituting six treatments with four replicates. Weight gain, concentrate intake, dry matter intake and feed:gain ratio were evaluated. For weight gain, differences (P <.05) for weight at 29, 36 and 50 days of age were observed, however, there were no differences for average daily gain (P>.05). There were also no differences (P>.05) for concentrate intake, dry matter intake and feed:gain ratio. In the second part of the experiment, three animals of each treatment were slaughtered, to evaluate the dietary effect on the organs performance, the intestines length, the empty body weight and the animals carcass chemical composition. For empty body weight and metabolic weight, there was significant effect (P<.05) for the used sources. For the animals carcass chemical composition, there were no significant differences among the sources or the zeranol use. For the organs weight, the omasum was influenced by the zeranol, occurring significant interaction for the mesenterium weight. The kidneys, lung and spleen weights were influenced by the fed dietary type. For the intestines length, just the thin intestine was influenced by the diet. It was concluded that the cow fermented colostrum and the soybean oil use come are viable alternative in the of pre-ruminant goats feeding, however, the use of the zeranol showed no results that justify its use for these animals.