La Biometrogie cutanée est un nouveau spectre de différentes méthodes d'évaluation qui permettent de mesurer les différents paramètres de la peau même en cas des petites modifications. Dans la première partie de nos travaux, nous avons étudié différentes capacités de ces méthodes afin de les corréler avec d'autres données cliniques et histologiques. Ces données nous ont encouragés à découvrir de plus en plus la pigmentation de la peau qui est un peu plus compliquée que les autres paramètres. Par conséquent, sur la deuxième partie, nous nous sommes concentrés sur la biologie cutanée en particulier sur la morphologie et l'activité des mélanocytes. Les travaux de cette thèse ont porté sur les effets de la phénytoïne sur les mélanocytes humains afin de savoir si cette molécule peut être un traitement efficace pour le vitiligo. Dans un premier temps, nous avons évalué deux concepts : tout d'abord et pour la première fois, les effets in vitro des différentes concentrations de phénytoïne sur la morphologie et l'activité des mélanocytes humains, et parallèlement, nous avons mis en place la méthode du transfert de mélanosomes sur les mélanocytes et des kératinocytes humains. Dans un deuxième temps, nous avons procédé à l'évaluation in vivo de la forme topique à différentes concentrations à l'aide d'une étude in vivo sur des cochons d'Inde noirs. Notre travail constitue une étape clé dans la compréhension des mécanismes d'action de la phénytoïne sur les mélanocytes humains qui contribuerait à l'amélioration des pratiques cliniques et donc à la qualité de vie des patients souffrant de troubles pigmentaire. / Cutaneaous Biometrogy is a new vast spectrum of measuring methods wich provide the possibility to measure the différents parameteres of the skin even in the case of small changes. In the first part of this work, we studied various capabilities of these methods in order to correlates them with other clinical and histological data. These data encouraged us to discover more and more the skin pigmentation which is a little more complicated than the other parameters.So, on the second part, we concentrated more on cutaneaous biology particularily on the melanocytes morphology and activity.The work of this thesis focused on the effects of phenytoin on human melanocytes in order to know if this molecule can be eventually an available treatment for vitiligo.Fisrt, we evaluated two concepts : for the first time we evaluated the in vitro effects of the different concentrations of phenytoin on the morphology and activity of human melanocytes and, in parallel, we implemented the method of transfert of melanosomes on human melanocytes and keratinocytes.In a second step, the topical form of phenytoin at different concentrations was evaluated through an in vivo study on black ginea pigs.Our work is a key step on understanding the mechanisms of avtion of phenytoin on human melanocytes which would contribute to the improvement of clinical practices and therefore to the quality of life of patients suffering from depilatory disorders.

Doctor of Philosophy / Department of Animal Sciences and Industry / Bradley J. Johnson / We used a total of fifty-nine gilts (BW=137.7 kg) from three different breeding groups were used to assess the effects of feeding L-carnitine during gestation on gilt growth characteristics, gilt and fetal blood metabolites, litter characteristics, and IGF axis components in fetal hepatic and skeletal muscle, maternal uterine and chorioallantois tissues, and porcine embryonic myoblasts collected from fetuses. Experimental treatments were arranged in a 2 × 3 factorial arrangement with main effects of L-carnitine and day of gestation. Gilts were fed a constant feed allowance of 1.75 kg/d and a top-dress containing either 0 or 50 ppm of L-carnitine starting on the first day of breeding through the allotted gestation length (40, 55, or 70). No differences (P > 0.16) were observed for BW or estimated protein or fat mass at any gestation length. Gilts fed L-carnitine tended to have greater (P = 0.10) backfat at d 40 and were numerically heavier at d 70 compared to control gilts. No differences (P > 0.77) were observed in circulating total and free carnitine at breeding, but concentrations increased (P < 0.01) as gestation length increased for the gilts fed L-carnitine compared to those fed the control diet. Fetuses from the gilts fed L-carnitine tended to be heavier (P = 0.06) and fetal circulating IGF-II lower (P = 0.09) at day 70 compared to the fetuses from the control gilts. Insulin-like growth factor-I (IGF-I) mRNA was lower (P = 0.05) in fetal hepatic tissue in fetuses collected from gilts fed supplemental L-carnitine. Insulin-like growth factor binding protein-3 (IGFBP-3; P = 0.05) and IGFBP-5 mRNA increased (P = 0.01) and IGF-I mRNA numerically increased (P = 0.16) in the endometrium of gilts supplemented with L-carnitine. At d 55 or 70 gestation, fetuses were removed for collection of porcine embryonic myoblasts (PEM) from the semitendinosus. There were no treatment differences (P > 0.10) for the expression of IGF-I, IGF-II or IGFBP-5 mRNA levels. However, PEM isolated from fetuses collected from gilts fed L-carnitine had lower (P = 0.08) IGFBP-3 mRNA levels compared to the controls. Myoblasts isolated from fetuses obtained from gilts fed L-carnitine had greater (P = 0.09; 8.8 %) 5.1H11 monoclonal antibody attachment after 72 h in culture. Although not significant (P = 0.20), the total number of PEM in the S phase of the cell cycle was 4.7 % greater in PEM collected from fetuses from gilts fed L-carnitine compared to the controls. This study shows L-carnitine supplementation to gestating gilts has beneficial effects on average fetal weight, due in part to changes in the expression of the IGF axis at the fetal-maternal interface in swine. These changes in the IGF axis play a fundamental role in porcine fetal growth and development due to enhanced proliferation and delayed differentiation of PEM.

Depuis plusieurs années, des projets d'intégration de données omiques se sont développés, notamment avec objectif de participer à la description fine de caractères complexes d'intérêt socio-économique. Dans ce contexte, l'objectif de cette thèse est de combiner différentes données omiques hétérogènes afin de mieux décrire et comprendre le dernier tiers de gestation chez le porc, période influençant la mortinatalité porcine. Durant cette thèse, nous avons identifié les bases moléculaires et cellulaires sous-jacentes de la fin de gestation, en particulier au niveau du muscle squelettique. Ce tissu est en effet déterminant à la naissance car impliqué dans l'efficacité de plusieurs fonctions physiologiques comme la thermorégulation et la capacité à se déplacer. Au niveau du plan expérimental, les tissus analysés proviennent de foetus prélevés à 90 et 110 jours de gestation (naissance à 114 jours), issus de deux lignées extrêmes pour la mortalité à la naissance, Large White et Meishan, et des deux croisements réciproques. Au travers l'application de plusieurs études statistiques et computationnelles (analyses multidimensionnelles, inférence de réseaux, clustering et intégration de données), nous avons montré l'existence de mécanismes biologiques régulant la maturité musculaire chez les porcelets, mais également chez d'autres espèces d'intérêt agronomique (bovin et mouton). Quelques gènes et protéines ont été identifiées comme étant fortement liées à la mise en place du métabolisme énergétique musculaire durant le dernier tiers de gestation. Les porcelets ayant une immaturité du métabolisme musculaire seraient sujets à un plus fort risque de mortalité à la naissance. Un second volet de cette thèse concerne l'imputation de données manquantes (tout un groupe de variables pour un individu) dans les méthodes d'analyses multidimensionnelles, comme l'analyse factorielle multiple (AFM) (ou multiple factor analysis (MFA)). Dans notre contexte, l'AFM fut particulièrement intéressante pour l'intégration de données d'un ensemble d'individus sur différents tissus (deux ou plus). Afin de conserver ces individus manquants pour tout un groupe de variables, nous avons développé une méthode, appelée MI-MFA (multiple imputation - MFA), permettant l'estimation des composantes de l'AFM pour ces individus manquants. / Over the last decades, some omics data integration studies have been developed to participate in the detailed description of complex traits with socio-economic interests. In this context, the aim of the thesis is to combine different heterogeneous omics data to better describe and understand the last third of gestation in pigs, period influencing the piglet mortality at birth. In the thesis, we better defined the molecular and cellular basis underlying the end of gestation, with a focus on the skeletal muscle. This tissue is specially involved in the efficiency of several physiological functions, such as thermoregulation and motor functions. According to the experimental design, tissues were collected at two days of gestation (90 or 110 days of gestation) from four fetal genotypes. These genotypes consisted in two extreme breeds for mortality at birth (Meishan and Large White) and two reciprocal crosses. Through statistical and computational analyses (descriptive analyses, network inference, clustering and biological data integration), we highlighted some biological mechanisms regulating the maturation process in pigs, but also in other livestock species (cattle and sheep). Some genes and proteins were identified as being highly involved in the muscle energy metabolism. Piglets with a muscular metabolism immaturity would be associated with a higher risk of mortality at birth. A second aspect of the thesis was the imputation of missing individual row values in the multidimensional statistical method framework, such as the multiple factor analysis (MFA). In our context, MFA was particularly interesting in integrating data coming from the same individuals on different tissues (two or more). To avoid missing individual row values, we developed a method, called MI-MFA (multiple imputation - MFA), allowing the estimation of the MFA components for these missing individuals.

Made available in DSpace on 2016-05-02T13:55:30Z (GMT). No. of bitstreams: 1 OswaldoHenriqueBarolliReis-Dissertacao.pdf: 707277 bytes, checksum: 3b71c133dbad0b57929d4b255619719c (MD5) Previous issue date: 2011-02-23 / In swine culture, the study of mycology contributes to food security in the understanding of various diseases. A knowledge of fungal agents on the skin of healthy animals and surface pathologies is important for effective control of the potential risks of infection, especially when associated with confined species, where the imbalance in the microbiota of the environment can be crucial in the interactions between environment and animals. The objective was to investigate the in vitro antimicrobial activity of essential oil of Rosa alba L. petals on microbial growth of yeasts Candida albicans (ATCC 10231) and filamentous fungi that causes opportunistic infectious processes in the skin of pigs, such as Microsporum nanum (ATCC 11832), Trichophyton mentagrophytes (ATCC 9533), Trichophyton rubrum (ATCC 10218) and Trichophyton tonsurans (ATCC 10870). The oils were extracted by the process of hydrodistillation in Clevenger apparatus, using fresh white rose petals. Microbiological tests were performed by the agar diffusion method (pour-plate) with Sabouraud-dextrose. Wells 5 mm in diameter, produced in the agar, were filled with 15 &#956;L of essential oil and the plates containing filamentous fungi were incubated in an inverted position at 28 °C for 10 days and the plates containing yeast were incubated at 30 °C for 48 hours. After this time, the inhibition circles were measured. The data were subjected to analysis of variance and means were compared by Tukey`s test at 5% probability. It was found that the activity of essential oil under study is significant in relation to the growth of microbial species tested. / Na suinocultura, o estudo da micologia contribui tanto na segurança alimentar como no entendimento de diversas patologias. O conhecimento dos agentes fúngicos presentes na pele de animais saudáveis e com patologias superficiais é importante para um controle eficaz dos riscos potenciais de infecção, principalmente quando associados a espécies confinadas, onde o desequilíbrio da microbiota ambiental pode ser decisivo nas interações entre o ambiente e os animais. Objetivou-se investigar a atividade antimicrobiana in vitro do óleo essencial de pétalas de Rosa alba L. sobre o crescimento microbiano de leveduras Candida albicans (ATCC 10231) e fungos filamentosos causadores de processos infecciosos oportunistas na pele de suínos, tais como Microsporum nanum (ATCC 11832), Trichophyton mentagrophytes (ATCC 9533), Trichophyton rubrum (ATCC 10218) e Trichophyton tonsurans (ATCC 10870). Os óleos foram extraídos pelo processo de hidrodestilação em aparelho de Clevenger, utilizando partes frescas das pétalas de rosa branca. Os testes microbiológicos foram realizados pelo método de difusão em ágar ( pour-plate ) com meio de cultura ágar Sabouraud-dextrose. Poços de 5 mm de diâmetro, produzidos no ágar, foram preenchidos com 15 &#956;L do óleo essencial e, em seguida, as placas contendo os fungos filamentosos foram incubadas de forma invertida, a 28 ºC por 10 dias; e as placas contendo a levedura foram incubadas a 30 ºC por 48 horas. Passado este tempo, os halos de inibição tiveram seus diâmetros medidos. Os dados obtidos foram submetidos à análise de variância e as médias comparadas através do teste de Tukey ao nível de 5% de probabilidade. Verificou-se que a atividade do óleo essencial em estudo é significativa em relação ao crescimento das espécies microbianas testadas.

O objetivo deste trabalho foi avaliar os efeitos de níveis de nucleotídeos nas dietas de leitões recém-desmamados sobre o desempenho, a morfometria de órgãos e a histologia do epitélio intestinal. Foram utilizados 144 leitões desmamados aos 21 dias de idade e com peso médio inicial de 5,80 ± 0,16 kg em um experimento inteiramente ao acaso, com seis tratamentos, seis repetições por tratamento e quatro animais por baia (unidade experimental). Os tratamentos foram: Am (antimicrobiano) dieta basal com inclusão de 40 ppm de sulfato de colistina, assim como dieta basal com 0, 150, 300, 450 e 600 ppm de nucleotídeos. Ao final do experimento, foi abatido um animal de cada baia para coleta das amostras para avaliação da morfometria de órgãos e da histologia do epitélio intestinal. No período de 1 a 14 dias de experimentação, houve piora linear (P<0,01) das variáveis de desempenho, enquanto que no período total de 34 dias, houve redução linear (P=0,03) do peso final dos animais com o aumento dos níveis de nucleotídeos na dieta. Os leitões alimentados com a dieta com o antibiótico colistina apresentaram maior (P<0,01) comprimento do intestino delgado e menor (P<0,01) relação altura de vilosidade:profundidade de cripta (AV:PC) no duodeno do que aqueles que receberam nucleotídeos. Foi observado, também, aumento linear (P<0,01) no peso relativo do baço, assim como redução linear (P<0,01) da relação AV:PC e redução linear (P<0,01) da profundidade de cripta no duodeno dos animais em função da adição de nucleotídeos na dieta. Assim, embora os níveis de até 600 ppm de nucleotídeos em dietas complexas de leitões recém-desmamados não tenham proporcionado melhora no desempenho, eles acarretaram benefícios à morfometria de órgãos e à histologia do epitélio intestinal dos animais. / The purpose of this work was to evaluate the effects of dietary nucleotide levels on performance, organs morphometry and intestinal histology of weanling pigs. One hundred and forty-four pigs weaned at 21 days of age and 5.80 ± .16 kg initial live weight were used a completely randomized design experiment with six treatments, six replications per treatment and four animals per pen (experimental unit). The treatments were: Am antimicrobial: basal diet with 40 ppm of colistin sulfate and basal diet with 0, 150, 300, 450 and 600 ppm of nucleotides. At the end of experimental period, an animal of each pen was slaughtered to evaluate of organs morphometry and intestinal epithelium histology. For the 1 to 14 days of experimental period, dietary nucleotides depressed linearly (P<.01) performance traits, while for the total experimental period of 34 days linear reduction of final live weight (P=.03) was observed with added nucleotides. Pigs fed diet with colistin showed greater (P<.01) length of small intestine and lower (P<.01) villus height:crypt depth ratio (AV:PC) of duodenum than those fed nucleotides. Also, a linear increase (P<.01) of relative weight of spleen, as well as a linear reduction (P<.01) of AV:PC and linear reduction (P<.01) of crypt depth of duodenum were observed with added dietary nucleotides. Therefore, although added nucleotides up to 600 ppm in complex diet did not improve performance, they showed some beneficial effects on organs morphometry and intestinal epithelium histology of weanling pigs.

O objetivo do trabalho foi estudar os efeitos de probióticos, prebióticos e extratos vegetais como alternativas ao uso de agentes antimicrobianos como promotores do crescimento de leitões recém-desmamados, por meio da morfologia de órgãos, frequência de diarréia, microbiologia intestinal, histologia de epitélio intestinal, digestibilidade dos nutrientes e desempenho. Foram realizados dois experimentos em blocos casualizados, nos quais foram testados cinco tratamentos: controle - ração basal; antimicrobiano - basal com 50 ppm de bacitracina de zinco + 50 ppm de olaquindox; probiótico - basal com 1300 ppm de probiótico à base de Bacillus subtilis e Bacillus licheniformis; prebiótico - basal com 3000 ppm de mananoligossacarídeo; extrato vegetal – basal com 500 ppm de extrato vegetal (alho, cravo, canela, pimenta, tomilho, cinamaldeído e eugenol). No Experimento I utilizaram-se 40 leitões desmamados aos 21 dias de idade, distribuídos a 20 gaiolas suspensas com dois leitões (um macho castrado e uma fêmea) em cada gaiola (unidade experimental) e quatro repetições por tratamento. Durante a última semana do período experimental foi realizado o ensaio de digestibilidade dos nutrientes, por meio da metodologia da coleta parcial de fezes, tendo o óxido crômico (Cr2O3) como marcador. Ao 35º dia, um animal de cada unidade experimental foi abatido para a coleta de dados referentes à morfometria de órgãos, de amostras do epitélio intestinal para análise histológica e de amostras do conteúdo do jejuno/íleo para análise microbiológica. No Experimento II, foram utilizados 120 leitões desmamados aos 21 dias de idade, alocados em 60 baias com dois animais (um macho castrado e uma fêmea) por baia (unidade experimental), perfazendo 12 repetições por tratamento. As variáveis de consumo diário de ração, ganho diário de peso e conversão alimentar foram coletadas e analisadas nos períodos de 1-14, 15-35 e 1-35 dias de experimentação. Também foi avaliada a frequência de diarréia. Apesar da diferença não ser significativa (P>0,05), em valores numéricos, os agentes antimicrobianos melhoraram o ganho diário de peso nos períodos de 15-35 (+22%) e 1-35 (+21,4%) dias de experimentação. Também melhoraram (P<0,05) a digestibilidade aparente de nutrientes (P<0,05) em relação ao tratamento controle. O prebiótico mananoligossacarídeo foi a melhor alternativa aos antimicrobianos, proporcionando um ganho diário de peso, numericamente (P>0,05), 25,7% superior ao tratamento controle e equivalente ao tratamento antimicrobiano, nos primeiros 14 dias de experimentação. Entretanto, o prebiótico mananoligossacarídeo não melhorou a conversão alimentar (P>0,10). Nos períodos de 15-35 dias e no período total, o desempenho dos animais do tratamento prebiótico foi intermediário entre o tratamento controle e antimicrobiano. O probiótico não melhorou o desempenho (P>0,10) e não proporcionou nenhuma alteração nas outras variáveis estudadas. Os extratos vegetais aumentaram o peso relativo do pâncreas (P<0,05) e melhoraram a digestibilidade aparente da matéria seca da dieta (P<0,05). No entanto, os extratos vegetais não melhoraram (P>0,10) o desempenho dos leitões. / The purpose of this work was to evaluate the probiotics, prebiotics and herbal extracts as alternatives to antimicrobial agents as growth promoters of weanling pigs, based on the morphometry of organs, fecal score, intestinal microbiology, intestinal histology, nutrient digestibility and performance. Two randomized complete block design experiments were carried out to compare five treatments: control - basal diet; antimicrobial - basal diet plus Zn bacitracin and olaquindox (50 ppm of each); probiotic - basal diet plus 1300 ppm of probiotic (Bacillus subtilis and Bacillus licheniformis); prebiotic - basal diet plus 3000 ppm of mannanoligosaccharide; herbal extract - basal diet plus 500 ppm of herbal extract (garlic, clove, cinnamon, pepper, thyme, cinnamaldehyde and eugenol). In Experiment I, forty 21-d-weaned pigs were alloted to 20 suspended pens, with two pigs (a barrow and a gilt) per pen (experimental unit) and four replications per treatment. In the last week, the digestibility assay was carried out, using the method of the partial faeces collection, with the chromium oxide (Cr2O3) as fecal marker. On 35th day, an animal of each experimental unit was slaughtered to collect the morphometry data, the intestinal epithelium samples for histological analysis, and the content of the jejunum/ileum for microbiological analysis. In Experiment II, one hundred and twenty 21-d-weaned pigs were used, with 12 replications per treatment and two animals (a barrow and a gilt) per experimental unit. The daily feed intake, the average daily gain and the feed conversion were analyzed for 1-14, 15-35 and 1-35 days of experimental periods. The fecal score was also evaluated. Although not significant (P>.05), the antimicrobial agents improved the average daily gain of weanling pigs, during 1-35 (+21.4%) and 15-35 (+22%) days of experimental periods, compared to pigs fed control diet. The nutrient apparent digestibility of pigs fed antimicrobial were higher (P<.05) than of those fed control. The prebiotic mannanoligosaccharide showed to be the most efficient alternative to antimicrobials. The average daily gain of pigs fed mannanoligosaccharide was, in numerical values (P<.05), 25.7% higher than of those fed control (P>.05) and similar (P>.05) to the animals of the antimicrobial treatment, during 1-14 days of experimental period. However, the feed conversion was not improved (P>.10). During 15-35 and 1-35 days of experimental periods, the performance of animals fed mannanoligosaccharide was intermediate between of those fed control and antimicrobial diet. The probiotic did not improve the growth performance of pigs (P>.10) and did not cause any effect. The herbal extracts increased the relative weight of pancreas (P<.10) and improved the dry matter apparent digestibility of the diet (P<.05), compared to control treatment. However, the herbal extracts did not improve pig performance (P>.10) in any period.

Os antibióticos têm sido utilizados como aditivos na alimentação animal para aumentar o desempenho e manter a saúde dos animais, como suínos e frangos, destinadas à produção de alimentos de origem animal. No entanto, desde 2006, a Comunidade Europeia proibiu o uso de antibióticos para esse proposito devido ao desenvolvimento de resistência bacteriana aos antibióticos. Como resultado, várias alternativas foram estudadas e propostas para substituir os antibióticos utilizados na alimentação animal. Os óleos essenciais têm recebido considerável atenção devido às suas propriedades antimicrobianas. Portanto, o objetivo do presente trabalho foi avaliar in vitro a atividade antibacteriana dos óleos essenciais contra a microbiota patogênica e probiótica de ocorrência no trato gastrointestinal de suínos e aves, destinadas à produção de alimentos de origem animal. A atividade antibacteriana seletiva, a qual, significou uma alta atividade antibacteriana contra bactérias patogênicas e reduzida ou nenhuma atividade sobre bactérias probióticas, foi avaliada como característica fundamental dos óleos essenciais com alto desempenho. Esta característica foi avaliada nos óleos essenciais usados individualmente e em combinações binarias. Inicialmente, no Capítulo 2, uma triagem de vinte e oito óleos essenciais (OEs) através do método de difusão em disco mostrou que Eucalyptusglobulus, E. exserta, Pimenta pseudocaryophylllus, Orange Oil Phase Essence, e CitrusTerpens (Os dois últimos OEs foram subprodutos do processamento da laranja para obtenção de suco) tiveram uma atividade antibacteriana seletiva sobre a bactéria patogênica Salmonella Enteritidis e a bactéria probiótica Lactobacillus plantarum. Numa fase posterior, esses cinco óleos foram avaliados individualmente e em misturas binárias, contra cinco bactérias patogênicas e três bactérias probióticas. Os melhores resultados foram observados quando os OEs foram avaliados isoladamente e não em misturas. Assim, Orange Oil Phase Essence e Citrus Terpens destacaram-se por ter a melhor atividade antibacteriana seletiva contra essas bactérias. No Capítulo 3, uma análise mais detalhada da atividade antibacteriana dos óleos essenciais foi realizada utilizando Orange Oil Phase Essence e a mistura composta pelos óleos de E. globulus e P. pseudocaryophyllus. Estes dois óleos foram selecionados com base nos resultados do Capitulo 2 e da disponibilidade de OEs em nosso estoque. Ambos, óleo e a mistura foram avaliados sobre a bactéria patogênica mais resistente, E. faecalis, e a bactéria probiótica menos resistente L. rhamnosus, como observado no Capitulo 2. A avaliação da Concentração Inibitória Mínima e Concentração Bactericida Mínima do Orange Oil Phase Essence e da mistura mostrou que não tiveram um efeito antibacteriano seletivo sobre E. faecalis e L. rhamnosus. Finalmente, no Capitulo 4, foi avaliada a atividade antibacteriana individual e combinada dos óleos de E. globulus e P. pseudocaryophyllus sobre E. faecalis e L. rhmanosus. Os resultados mostraram que a combinação destes dois OEs, avaliadas pelo método checkerboard, não potencializou a atividade antibacteriana seletiva dos dois OEs. Portanto, observou-se que o óleo de E. globulus isoladamente apresentou a melhor atividade antibacteriana seletiva contra E. faecalis e L. rhamnosus. Em conclusão, este trabalho permitiu identificar óleos essenciais com perfil antibacteriano seletivo para eles serem possíveis alternativas botânicas aos antibióticos utilizados na alimentação animal. / Antibiotics have been used in animal feed to maintain health and increase performance, as in the case of pigs and poultry intended for food production of animal origin. However, since 2006 the European Community has banned the use of antibiotics for this purpose due to the emergence and increase of antibiotic-resistant bacteria. As a result, several alternatives have been studied and proposed to substitute antibiotics used in animal feed. Essential oils have received considerable attention due to their antimicrobial properties. Therefore, the objective of this work was to evaluate in vitro the antibacterial activity of essential oils against pathogenic and probiotic bacteria that occur in the gastrointestinal tract of swine and poultry, intended for food production of animal origin. The selective antibacterial activity, which means high antibacterial activity on pathogenic bacteria and reduced or no activity on probiotic bacteria, was evaluated as a fundamental feature of the highest-performance essential oils. This feature was evaluated in essential oils used individually and in binary combinations. Initially, in Chapter 2, the screening of twenty-eight essential oils (EOs) by disk diffusion method showed that Eucalyptus globulus, E. exserta, Pimenta pseudocaryophylllus, Orange Oil Phase Essence, and Citrus Terpens (the last two EOs were by-products of orange juice production) had a selective antibacterial activity against the pathogenic Salmonella Enteritidis and probiotic Lactobacillus plantarum. At a later stage those five oils were evaluated, individually and in binary blends, against five pathogenic bacteria and three probiotic bacteria. Better results were observed when the EOs were checked alone and not in blends. Orange Oil Phase Essence and Citrus Terpens stood out for having the two best selective antibacterial activities against those bacteria. In Chapter 3, a more detailed analysis of essential oil antibacterial activities was perfomed using Orange Oil Phase Essence and the blend composed of E. globulus and P. pseudocaryophyllus. These two oils were selected based on the results of Chapter 2 and from the availability of our EO stock. Both oil and blend were checked on the most resistant pathogenic bacterium, E. faecalis, and on the less resistant probiotic bacterium of the Lactobacillus genus, L. rhamnosus, as observed in Chapter 2. The evaluation of Minimal Inhibitory Concentration and Minimal Bactericidal Concentration for Orange Oil Phase Essence and the blend showed that there was not a selective antibacterial effect against E. faecalis and L. rhamnosus. Finally, in Chapter 4, the individual and combined antibacterial activities of E. globulus and P. pseudocaryophyllus essential oils on E. faecalis and L. rhmanosus were evaluated. The results showed that the combination of two EOs evaluated by checkerboard method did not potentiate the selective antibacterial activity of the two EOs. Therefore, it was observed that the E. globulus essential oil alone had the best selective antibacterial activity against E. faecalis and L. rhamnosus. In conclusion, this work enabled the identification of essential oils with selective antibacterial profile that can become possible botanical alternatives to antibiotics used in animal feed.

O presente trabalho de pesquisa foi conduzido no período de outubro de 2007 a março de 2008 na Estação de Avaliação de Suínos em Tanquinho, Piracicaba SP, no Frigorífico BRESSIANI®, localizado na cidade de Capivari SP, e as avaliações bioquímicas, químicas, quantidade e qualidade de carne foram realizadas no mês de março a abril de 2008, no Centro de Pesquisa e Desenvolvimento de Carnes do Instituto de Tecnologia de Alimentos, localizado na cidade de Campinas SP. Neste experimento foram testados métodos de castração de suínos objetivando avaliar a incidência do odor de macho inteiro, causado pela androstenona e escatol, presentes no tecido adiposo dos suínos machos, composição corporal e da carcaça. Entre os métodos de castração de suínos machos existentes, foram avaliados o cirúrgico e imunológico. O primeiro consistiu na remoção cirúrgica das gônadas do leitão entre 3 a 5 dias de vida e o segundo na aplicação de duas doses da forma modificada do Fator de Liberação das Gonadotropinas (GnRF) em um sistema coadjuvante de baixa reatividade, sendo a primeira dose aplicada nos animais com 15 semanas e a segunda dose com 19 semanas de idade. Após duas semanas da segunda dose foram coletadas amostras de sangue para avaliação de testosterona através de radioimunoensaio. No momento do abate foi realizada a coleta de parte do tecido adiposo para análise de androstenona e escatol. Nas meias carcaças esquerdas foram avaliadas a qualidade de carne (pH, temperatura, capacidade de retenção de água, perda por exsudação e cor) e a quantidade (medidas lineares das carcaças e rendimentos dos cortes anatômicos). A vacinação dos animais contra o GnRF demonstrou sua eficiência em controlar os compostos responsáveis pelo odor sexual (androstenona e escatol) e em manter os níveis de testosterona comparáveis aos animais castrados cirurgicamente. A imunocastração melhorou o desempenho zootécnico e contribuiu para aumentar a quantidade de carne por animal (4,84 kg), diminuir a de gordura (1, 54 kg) e acrescentar mais carne nos cortes de maior valor comercial, como pernil (1060 g, p < 0,05), carre(400 g, p > 0,05), a barriga (840 g, p < 0,05) e paleta (1460 g, p < 0,05), que representa uma vantagem econômica para a indústria da carne, pois atende os mercados de carne fresca e de produtos industrializados (cozidos e embutidos). Em relação à avaliação sensorial foram constatadas diferenças significativas (p < 0,05) para todos os atributos sensoriais avaliados em favor dos suínos imunocastrados quando comparados com os castrados cirurgicamente. Quanto à preferência, os bifes cozidos de lombo dos suínos imunocastrados obtiveram melhor preferência (66%) em comparação com os castrados cirurgicamente (34%). A intenção de compra também foi em favor dos suínos imunocastrados e refletiu os resultados dos testes de preferência e de aceitação. A maioria dos consumidores (74,8%) provavelmente (20,2%) ou certamente (54,6%) compraria a carne dos suínos imunocastrados e somente 58,4% dos consumidores provavelmente (25,2%) ou certamente (33,2%) compraria a carne dos suínos castrados cirurgicamente. A gordura total encontrada no lombo nesse experimento foi 14.67 e 12.67 g/100g para o grupo dos suínos castrados cirurgicamente e imunocastrados, respectivamente. O rendimento de cocção e os valores da força máxima de cisalhamento (Warner - Bratzler) dos suínos castrados cirurgicamente e imunocastrados não foram estatisticamente diferentes (p > 0,05) enquanto que a cor instrumental (L*, a* e b*) apresentou diferença estatística (p < 0,05) na sua composição para os tratamentos estudados. Os resultados desse trabalho permitem dizer que a imunocastração demonstrou ser eficiente em prevenir o odor sexual bem como em melhorar o desempenho zootécnico e qualidade de carcaça quando comparada com a castração cirúrgica. Quanto à qualidade de carne dos suínos imunocastrados ficou evidenciado que essa tecnologia pode melhorar os atributos sensoriais e outras características de qualidade realizadas nessa pesquisa. / The present study was carried out from October 2007 to March 2008 at the Pig Evaluation Station in Tanquinho, located in Piracicaba city, São Paulo state, in the BRESSIANI® abattoir, located in Capivai, city, São Paulo state, and the biochemical, chemical, meat quantity and meat quality assessments were carried in March 2008, at the Meat Center for Research and Development in the Institute of Food Technology, located in Campinas city, São Paulo state. In this experiment were tested castration methods applied in pigs to evaluate the incidence of the boar taint, caused by androstenone and skatole located in the fat of boars, carcass composition and meat quality. The castration methods tested were physical and immunological. In the first one pig gonads were removed physically at the age 3 to 5 days and the last one the pigs were immunocastrated (two doses, 15 and 19 weeks of age) against the modified factor gonadotropin-releasing (GnRF). After two weeks of the second dose blood samples were taken to evaluate testosterone using radioimmunoassay. At the slaughter level a portion of backfat was collected to assess androstenone and skatole. Meat quality (pH, temperature, water holding capacity, loss of weeping and color) and quantity (linear measurements of carcasses and anatomical cuts yield) were performed at left carcass side. Vaccination of animals against GnRF demonstrated its effectiveness in controlling the boar taint compounds (androstenone and skatole) and to maintain the levels of testosterone comparable to physically castrated animals. The immunocastration improved growth performance and contributed to increase the total meat quantity per animal (4.84 kg), reducing the fat (1, 54 kg) as well as added more meat in cuts of higher commercial value, such as ham (1060 g, p < 0.05), loin (400 g, p > 0.05), belly (840 g, p < 0.05) and shoulder (1460 g, p < 0.05), which represents an economic advantage for the meat industry since reaches booths markets fresh and processed meats products (sausage and cooked). Regarding the sensory evaluation were found significant differences (p < 0.05) for all sensory attributes evaluated in favor ofimmunocastrated pigs when compared with physically castrated. The preference test applied to cooked sirloin steak from immunocastrated pigs indicated better preference (66%) compared with physically castrated (34%). The panelists intent to purchase was also in favor of the immunocastrated treatment and confirmed the results from the preference and acceptance tests. The majority (74.8%) of the consumers probably (20.2%) or certainly (54.6%) would buy meat from the immunocastrated pigs compared to 58.4% of the consumers who would probably (25.2%) or certainly (33.2%) would buy meat from physically castrated pigs. Loin total fat content found in the experiment was 14.67 and 12.67 g/100g for the physical and immunocastrated groups respectively. The immunocastrated group contained 11.2% less fat than the surgical castrates. Cooking yields and peak Warner-Bratzler shear force values from physically castrated and immunocastrated pigs were not statistically different (P > 0.05) while statistically significant differences (p < 0.05) in the brightness (L*), redness (a*) and yellowness (b*) parameters were observed between the two treatments. From the above considerations it can be said that immunocastration shows very good potential for pre-venting boar taint. Pain and stress associated with physical castration can thus be avoided. It was also demonstrated that immunocastrated pigs improved some meat quality aspects evaluated. Sensory, total fat content and color were the main factors affected by immunocastration while cooking loss and instrumental tenderness had no remarkable changes. The consumers classified meat from immunocastrated pigs significantly better than physically castrated pigs as far as acceptability, preference and purchase intention are concerned. Thus, immunocastration results in production of animals with high meat quality in the carcass and still capitalizes on the growth, feed efficiency and carcass leanness of boars up to the point of immunocastration.

Hintergrund und Zielstellung: Im Zuge der Intensivierung der Schweinemast kommen Atemwegsinfektionen und deren klinische Auswirkungen auf die Tiere, in Verbindung mit möglichen Wechselwirkungen zu den Umgebungsfaktoren, eine entscheidende Bedeutung für die Tiergesundheit und Wirtschaftlichkeit der Betriebe zu. Daher war es Ziel dieser Arbeit, unter Feldbedingungen festzustellen, inwieweit Korrelationen zwischen Parametern des Stallklimas, dem Nachweis potentiell pathogener Infektionserreger des Respirationstraktes, klinischem Befund der Tiere, dem Lungenbefund am Schlachtband und den Leistungsfaktoren bestehen. Material und Methoden: Über den Zeitraum von einem Jahr wurden fünf Schweinemastbetriebe in Thüringen, mit Betriebsgrößen im Bereich von 2480 bis 22 000 Mastplätzen hinsichtlich Tiergesundheit, insbesondere der Atemwegsgesundheit und Stallklima untersucht. Betriebskenndaten über Stallbau, Belegungsdichte, Lüftungs- und Heiztechnik sowie Fütterungs, -Tränke- und Impfregime wurden aufgenommen. Für die Untersuchungen wurden pro Betrieb zwei Abteile ausgewählt. Dabei lagen die betriebsabhängigen Abteilgrößen zwischen 159 bis 751 Tieren. In diesen Abteilen wurden bei einer Stichprobengröße von zehn Tieren in zwei Mastperioden, Sommer und Winter, zu jeweils drei Zeitpunkten (Vormast, Mittelmast und Endmast) 840 Blutproben entnommen und bezüglich der lungenpathogenen Erreger PRRSV, PCV 2, SIV sowie APP untersucht. PRRSV und PCV 2 betreffend, wurde sowohl auf Virus mittels in house-PCR-Untersuchungen gepoolter Proben, als auch serologisch auf Antikörper mittels ELISA untersucht. Für den Erreger SIV erfolgte eine serologische Untersuchung auf Antikörper mittels HAH sowie für APP mittels ELISA. Des Weiteren kam es in den entsprechenden Abteilen zur Durchführung stallklimatischer Messungen, wobei die Stallklimaparameter Lufttemperatur, Relative Luftfeuchte, Luftgeschwindigkeit sowie die Beleuchtungsstärke mithilfe von Geräten der Firma Testo sowie die Konzentrationen von CO2, O2, NH3und H2S mithilfe eines Multiwarn-Mehrgasmessgeräts der Firma Dräger ermittelt wurden. Gleichzeitig erfolgte eine klinische Befunderhebung in den Gruppen, mit der Bildung eines Klinikscores mit Schwerpunkt Lungengesundheit, sowie die Aufnahme der Leistungsparameter: durchschnittliche tägliche Zunahme, Mortalität der Gruppe und dem Tierbehandlungsindex, aus denen ein Leistungsscore ermittelt wurde. Im Zuge der Schlachtung wurden 9921 Schlachtkörper, insbesondere das Geschlinge und die Lungen, makroskopisch beurteilt und daraus ein modifizierter Organboniturscore, auf Grundlage des Organbefundindex nach Blaha gebildet. Alle Score-Bewertungen erfolgten nach Punkten und gaben somit eine Aussage zum Tiergesundheitsstatus der Betriebe. Höhere Punktzahlen entsprachen einem schlechterem Gesundheits- bzw. Leistungsstatus. Somit konnten Wechselwirkungen zwischen den Ergebnissen der Blutuntersuchungen, den Stallklimaparametern, dem Klinik- und Leistungsscore sowie dem Organboniturscore mittels statistischer Auswertung hergestellt werden, wobei die einzelnen Bestände sowohl betriebsübergreifend als auch auf Bestandsebene untersucht wurden. Aufgrund durchgeführter Umbaumaßnahmen in drei Beständen wurde ein Vergleich alter und neuer Betriebsteile in die Betrachtungen mit einbezogen. Ergebnisse: Keiner der Betriebe wies eine vollständige Erregerfreiheit auf, was für die endemische Verbreitung der Infektionserreger, insbesondere PRRSV, PCV 2 sowie APP spricht. SIV wurde lediglich in einem Bestand nachgewiesen. Serokonversionen hinsichtlich PRRSV gingen signifikant mit vermehrtem Husten der Tiere (p = 0,027) und einer höheren klinischen Gesamtpunktzahl (p = 0,016) einher. Bei Virusnachweis zeigte sich mit p = 0,047 eine signifikant schlechtere Lungengesundheit. Hinsichtlich des Stallklimas bestand für die Parameter Temperatur, Relative Luftfeuchte, CO2 Gehalt sowie den Schadgasgehalten der Stallluft ein starker Bezug zu den unterschiedlichen Mastperioden Sommer und Winter, mit vorrangig schlechteren Werten in der Winterperiode. Diese Klimaparameter lagen oftmals über den Grenz- bzw. Regelwertbereichen, mit Auswirkungen auf die Atemwegsgesundheit. Mit Ausnahme des Parameters NH3 ergaben sich bestandsübergreifend mit p ≤ 0,01 signifikante Zusammenhänge zu den klinischen Befunden (höhere Teilpunkzahlen Lungengesundheit korrelieren mit ρ = 0,250 zum CO2-sowie mit ρ = 0,222 zum H2S-Gehalt der Stallluft, negativ mit ρ = −0,396 zur Temperatur) sowie NH3 inbegriffen (mit einer Korrelation von ρ = 0,401) zu den Pneumoniebefunden. Ebenfalls bestandsübergreifend zeigte sich ein signifikanter Zusammenhang zwischen der Infektion von APP (p = 0,038) sowie PRRSV (p = 0,001) und einer erhöhten Anzahl an Pleuritiden. Des Weiteren zeigten sich mit einer Signifikanz von p ≤ 0,01 in Verbindung mit verstärkten klinischen Symptomen auch signifikant mehr makroskopisch sichtbare Pneumonien bei der Schlachtung (Korrelation der Gesamtpunktzahl Klinik mit ρ = 0,297). Bestandsabhängig wurden beim Vorhandensein multipler Erregerspektren im Betrieb erhöhte Lungenbefunde ermittelt. Beziehungen der Klima- und Klinikparameter sowie der Lungenbefunde zu den Leistungsparametern, konnten nur in geringem Maße festgestellt werden. So wurden beispielsweise keine Korrelationen zwischen den Schadgasparametern und der durchschnittlichen Masttagszunahme festgestellt. Hier scheint der genetische Aspekt einen starken Einfluss zu besitzen. Schlussfolgerungen: Anhand der Untersuchungen konnten sowohl einzelbetrieblich als auch betriebsübergreifend Signifikanzen zwischen den einzelnen Parametern festgestellt werden. Die gegenseitige Beeinflussung und Abhängigkeit aller Faktoren ist jedoch immer herdenabhängig und variiert, je nach Bestandsdynamik.

Bei Anwendung reversibler Betäubungsmethoden erfolgt die Tötung von Schlachttieren durch Blutentzug. Findet dieser nicht oder nicht in ausreichendem Maße statt, besteht die Gefahr, dass die Tiere ihre Wahrnehmungs- und Empfindungsfähigkeit wiedererlangen, bevor sie weiteren Verarbeitungsschritten zugeführt werden. Dies ist aus Tierschutzsicht keinesfalls hinnehmbar und steht im Widerspruch sowohl zum Tierschutzschlachtrecht, als auch zu Verbrauchererwartungen im Hinblick auf einen tierschutzgerechten Schlachtablauf. Es ist deshalb dringend erforderlich den Todeseintritt jedes Einzeltieres zu verifizieren. Dies kann durch direkte Kontrolle der Schlachttiere (klinische Untersuchung, Prüfung der Hirnstammreflexe) nach Eintritt des Todes erfolgen oder bereits frühzeitiger, über eine Erfassung von Parametern, die eine Aussage über die Wahrscheinlichkeit des kurz bevorstehenden Todeseintritts erlauben (TROEGER 2011). Hierfür eignet sich insbesondere die Stichblutmenge, bzw. die Blutflussrate, da ab einer bestimmten Blutmenge pro Zeiteinheit vom kurz bevorstehenden Tod des Tieres ausgegangen werden kann (TROEGER 2010). Zielsetzung dieser Arbeit war zum einen, ein geeignetes Messsystem zur Erfassung des Entbluteverlaufs zu finden und dieses in das Stechkarussell zu integrieren, zum anderen sollte der Zusammenhang zwischen Stichblutmengenanstieg und Todeseintritt der Tiere untersucht werden. In einem ersten Schritt wurden vier verschiedene Messsysteme zur Erfassung des Entbluteverlaufs unter Zuhilfenahme eines Prototyps geprüft: Stichblutwaage (Fa. Bizerba), Füllstandsmesssonde (Fa. Sick), Infrarotkamera „thermoIMAGER TIM“ (Fa. Micro-Epsilon) und Infrarotsensoren „thermoMETER CSmicro“ (Fa. Micro-Epsilon). Als Messparameter und Maß für die Qualität der Entblutung bzw. des Entblutestichs, wurde die Steigung des Blutanstiegs im Blutauffangbehälter verwendet. Das Messsystem „Infrarotsensoren“ erwies sich als am besten geeignetes System und wurde nachfolgend in die Stechkarussellanlage des Partnerschlachthofes integriert und der Messvorgang automatisiert. Um den Zusammenhang zwischen Stichblutmengenanstieg und Todeseintritt der Schweine auf der Nachentblutestrecke zu untersuchen, wurde der Entbluteverlauf in den ersten Sekunden nach dem Stich sowohl mit dem Prototyp (Messsystem Waage) als auch mit dem integrierten IR-Sensor-Messsystem bei einer Stichprobe von 1034 (Prototyp) bzw. 1500 (integriertes System) Schweinen erfasst und anschließend der Eintritt des Hirntodes der Tiere klinisch überprüft (u.a. Untersuchung der Hirnstammreflexe). Im Rahmen eines Vorversuchs wurde eine Status quo-Erhebung von Stichblutmenge- und anstieg durchgeführt und die ersten fünf Sekunden nach dem Entblutestich als geeigneter Messzeitraum festgelegt. Es konnte gezeigt werden, dass der Anstieg der Entblutekurve in diesem Zeitraum annähernd linear verläuft, unabhängig von Lebendgewicht und Geschlecht der Tiere ist, und zudem eine gewisse Voraussage über den weiteren Verlauf der Entblutung bzw. die am Ende der Entblutung absolut entzogene Blutmenge zulässt. In der Hauptuntersuchung waren bei keinem Tier deutliche Anzeichen für Empfindungs- und Wahrnehmungsvermögen (gerichtete Augenbewegungen, Vokalisation) vorhanden. 3,6 Prozent (Prototyp-Untersuchung) bzw. 3,7 Prozent (integriertes IR-Sensor-Messsystem) der Tiere zeigten jedoch bei der Verifizierung des Todeseintritts noch bzw. wieder Reaktionen, die zumindest auf eine gewisse (Rest-)Hirnfunktion hindeuteten. Ein signifikanter Zusammenhang zwischen Steigung der Entblutekurve und nachfolgendem Todeseintritt der Tiere konnte weder in der Untersuchung mit dem Prototyp, noch in der Untersuchung mit dem integrierten IR-Sensor-Messsystem dargestellt werden. Die Ursache dafür wird in einer Beeinflussung der Ergebnisse durch tierindividuell unterschiedliche Betäubungstiefen vermutet (irreversibel betäubte Tiere, deren Entblutekurven eine geringe Steigung aufwiesen, die aber trotzdem keine Reaktionen auf der Nachentblutestrecke zeigten). Im Rahmen der vorliegenden Arbeit wurde ein vollständig automatisiertes, mit herkömmlichen Stechkarussellanlagen kompatibles, Entblutekontrollsystem auf Grundlage der Infrarotsensor-Technik entwickelt. Durch die Fokussierung auf die ersten fünf Sekunden nach Setzen des Entblutestichs, ist das Ergebnis bereits verfügbar, wenn sich die Tiere noch in der Reichweite des Stechpersonals befinden, wodurch eine sofortige Nachbearbeitung betroffener Schweine, ohne zusätzlichen Personaleinsatz, ermöglicht wird. Weiterhin erlaubt das berührungslos arbeitende System eine Einzeltierkontrolle, inklusive automatischer Dokumentation der Entblutedaten, bei der sowohl unzureichend als auch versehentlich überhaupt nicht entblutete Schweine erkannt werden. Die Entwicklung des automatisierten Entblutekontrollsystems trägt dazu bei, den Tierschutz bei der Schweineschlachtung zu verbessern und gesetzliche Vorgaben sowie Verbrauchererwartungen im Hinblick auf tierschutzgerechte Fleischgewinnung als Teil der Lebensmittelqualität zu erfüllen.