A remoção de microrganismos patogênicos através da desinfecção é uma necessidade para diminuir a incidência de doenças na população humana relacionadas com poluição fecal. A desinfecção sequencial tem grande potencial na remoção de microrganismos, quando comparada com a desinfecção convencional (um desinfetante). Neste trabalho buscou-se comparar a resistência de microrganismos indicadores de bactérias - E. coli e coliformes totais - e bactérias esporuladas - Clostridium perfringens quanto à desinfecção sequencial empregando cloro seguido de radiação ultravioleta e ozônio seguido de radiação ultravioleta; e à desinfecção convencional utilizando-se os mesmos desinfetantes aplicados individualmente em esgoto sanitário tratado previamente em reator UASB. Os ensaios foram realizados em batelada. As dosagens de cloro aplicadas foram de 10, 20 e 30 mg\'CL IND.2\'/L; de ozônio foram de 5,6; 11 e 16,5 mg\'O IND.3\'/L, ambos para os tempos de contato 10, 20 e 30 minutos. Na desinfecção sequencial com cloro foram aplicadas as doses de 1, 5 e 10 Wh/\'M POT.3\' de radiação UV; com ozônio, as doses de radiação foram variadas de 0,5 a 10 Wh/\'M POT.3\'. Na desinfecção sequencial de cloro-UV, foram removidos 2,5 e 5,2 log de C. perfringens e coliformes totais, respectivamente contra 1,5 e 4,2 log na desinfecção convencional para os mesmos microrganismos. Na desinfecção com cloro, a ordem decrescente de resistência foi: C. perfringens > coliformes totais > E. coli. Para ozônio seguido de UV, C. perfringens apresentou maior resistência e, em alguns ensaios, E. coli apresentou-se mais resistente que coliformes totais. A ação do ozônio mostrou-se notável para a melhoria da qualidade do esgoto tratado avaliada pela diminuição das concentrações de sólidos suspensos totais, sólidos totais, absorbância em comprimento de onda de 254 nm e da DQO, diferentemente do esgoto clorado no qual ocorreu o aumento nos valores destas variáveis físico-químicas. A ação da radiação ultravioleta foi potencializada quando aplicada sequencialmente ao cloro e ao ozônio. / The removal of pathogenic microorganisms through disinfection is a necessity to decrease the incidence of diseases in the human population related to fecal pollution. Sequential disinfection has a great potential on the removal of microorganisms when compared to conventional disinfection (single disinfectant). This work compared the resistance of indicators microorganisms of bacteria - E. coli and total coliforms - and spore-forming bacteria - Clostridium perfringens concerning the sequential and conventional disinfections. The sequential disinfection initially employed chlorine followed by ultraviolet radiation and ozone followed by ultraviolet radiation. The conventional disinfection utilized the same disinfectants individually applied in sanitary sewage previously treated in UASB reactor. The tests were made in batch. The chlorine dosages applied were 10, 20 and 30 mg\'CL IND.2\'/L; ozone were 5,6; 11 and 16,5 mg\'O IND.3\'/L, both for the contact times 10, 20 and 30 minutes. In the sequential disinfection with chlorine dosages of 1, 5 and 10 Wh/\'M POT.3\' of UV radiation were applied; with ozone, the dosages of radiation varied from 0,5 to 10 Wh/\'M POT.3\'. In the sequential disinfection with UV-chlorine were inactived 2,5 and 5,2 log of C. perfringens and total coliforms respectively, against 1,5 and 4,2 log in the conventional disinfection for the same microorganisms. In the chlorine disinfection, the decreasing resistance order was: C. perfringens > total coliforms > E. coli. In the ozone disinfection followed by UV, C. perfringens manifested more resistance and, in some assays, E. coli was more resistant than total coliforms. The action of ozone was outstanding for the improvement of treated sewage quality assessed by the decreasing of total suspended solids concentration, total solids absorbance at a wavelength of 254 nm and of DQO, differently from the chlorine sewage in which occurred an increase of the values of the physical-chemical variables. The ultraviolet radiation action was potentiated when sequentially applied to chlorine and to ozone.

Este trabalho tem como finalidade obter condições de recuperar e purificar a pró-toxina épsilon e uma protease produzida pelo Clostridium perfringens através do uso da técnica de extração líquido-líquido em sistema de duas fases aquosas (SDFA) para utilização na produção de vacinas. A aplicação do sistema de duas fases aquosas é proposta como alternativa para a purificação, pois permite a separação e análise de partículas biológicas. Esta técnica é aconselhável para purificação em larga escala pela possibilidade de partição seletiva com altos rendimentos, além de apresentar uma boa relação custo-benefício. Foram construídas as curvas binodais que foram utilizadas para análise da composição dos sistemas de duas fases aquosas formados por polietileno glicol e citrato de sódio, como também a extração e recuperação da pró-toxina épsilon e da protease produzidas por Clostridium perfringens. As curvas binodais foram construídas utilizando PEG 400, 550, 1000, 1500, 3350 e 8000 em diferentes valores de pH (6,0; 6,5; 7,0; 7,5 e 8,0) e água para formação do sistema. Também foram construídas curvas na presença de caldo clarificado em substituição à água. Foi avaliada ainda a estabilidade da pró-toxina épsilon, antes da extração, e da protease antes e após a extração, frente às variações de pH e temperatura. Um planejamento experimental ('2pot.8-3') foi usado para avaliar a influência das variáveis concentração e massa molar do PEG, concentração de citrato, pH, concentração de NaCl, fator de diluição do extrato, temperatura e massa total do sistema na extração da pró-toxina épsilon. A partição de uma protease presente no meio fermentado de C. perfringens foi estudada através do uso de três planejamentos experimentais completos (dois do tipo '2pot.4' e um do tipo '2pot.3') que avaliaram a influência das variáveis concentração e massa molar do PEG, concentração de citrato e pH. Três variáveis-resposta foram obtidas (aumento de pureza, coeficiente de partição e recuperação da enzima). Os resultados atingidos foram: coeficiente de partição de 0,57, aumento de pureza de 4,2 com uma recuperação de 131% da atividade enzimática na fase superior do sistema. O sistema de extração que proporcionou as melhores condições de extração foi constituído por: PEG 10000 (g/mol) e concentração de 22% (m/m), concentração de citrato de 8% (m/m) e pH 8,5. A protease permaneceu estável (durante 48 h), mesmo após a extração, nas temperaturas de 5°C e 25°C e nos valores de pH de 6,0 a 9,0. / The purpose of this work is to obtain best conditions of recovery and purification of proteins (epsilon prototoxin and a protease) produced by Clostridium perfringens through the use of the liquid-liquid extraction by aqueous two-phases systems (ATPS). The application of these systems is proposed as alternative for protein purification, because it allows the separation and analysis of biological particles. This technique is advisable process purification applied to large scale since it provides a selective partition with high yields, and good cost-benefit ratio. The binodal curves were constructed and used to determine the system composition based on polyethylene glicol and citrate concentrations. The binodal curves were built by using PEG 400, 550, 1000, 1500, 3350 and 8000 g/mol at different pH values (6.0; 6.5; 7.0; 7.5 and 8.0). The curves were, initially, built in the presence of water and, later, with clarified fermented broth. The differences in the curve profiles (water versus broth) helped to explain the phase separation behaviour. The stability, as a function of pH and temperature, of the epsilon prototoxin was evaluated before the extraction, while the stability of the protease was evaluated before and after the extraction. An experimental design ('2pot.8-3') was used to evaluate the influence of the following variables on epsilon prototoxin extraction: concentration and molar mass of PEG, citrate concentration and NaCl concentrations, pH, dilution factor of the extract, temperature and total mass of the system. However, the partition of the protease was studied through the use of the three full different experimental designs (two of the type '2pot.4', and one of the type '2pot.3') that evaluated the influence of the following variables: concentration and molar mass of PEG, citrate concentration and pH. Three parameter responses were obtained: purification factor; coefficient partition; and recovery yield of the enzyme. The results were satisfactory: partition coefficient = 0.57, purification factor = 4.2; yield = 131%. The extraction conditions which provided the best results were: molar mass of PEG 10000 (g/mol); concentration of PEG of 22% (w/w); concentration of citrate of 8% (w/w); and pH 8.5. The protease was stable (during 48h), even after the extraction, in the temperatures of 5°C and 25°C, and at pH values of from 6.0 to 9.0.

Clostridium perfringens causa enterite necrótica aviária devido à produção de toxinas que lesam o intestino. Neste estudo, de 94 amostras nove apresentaram C. perfringens, totalizando 22 isolados. Todos exceto um isolado, possuíram os genes nanI (95%) e/ou nanJ (81%), e 19/22, mostraram atividade neuraminidase em hemácias de frango. A atividade hemaglutinante foi observada em poucos isolados (26%). Todos os isolados foram plc positivos (toxina α), sendo classificados como tipo A. Sete isolados (31,8%) abrigaram o gene tpeL que codifica a toxina TpeL. Isolados tpeL+ mostraram efeito citotóxico característico da ação desta toxina. Alguns isolados mostraram capacidade de aderir e invadir células Vero. A maioria dos isolados foi resistente à sulfaquinoxalina (100%), cefalexina (95%) e eritromicina (95%) e sensíveis (100%) à cefoxitina, amoxicilina, enrofloxacina, amoxicilina-ácido clavulânico, penicilina-estreptomicina, cloranfenicol e metronidazol. Todos os isolados foram agrupados geneticamente em sete clusters, apresentando se como um grupo heterogêneo. / Clostridium perfringens cause avian necrotic enteritis due to production of toxins that damage the intestine. In this study, nine out of 94 samples had C. perfringens, totaling 22 isolates. All the isolates with exception of one, possessed the genes nanI (95 %) and/or nanJ (81 %), and 19/22 showed neuraminidase activity in chicken erythrocytes. The hemagglutinating activity was observed in a few isolates (26 %). All isolates were plc positive (toxin α) being classified as type A. Seven isolates (31.8%) harbored tpeL gene encoding the toxin TpeL. TpeL + isolates showed characteristic cytotoxic effect of the action of this toxin. Some isolates showed ability to adhere and invade Hep-2 cells. Most of the isolates were resistant sulphaquinoxaline (100%), cephalexin (95%) and erythromycin (95%) and sensitivity (100%) to cefoxitin, amoxicillin, enrofloxacin, amoxicillin - clavulanic acid, penicillin -streptomycin, chloramphenicol and metronidazole. All isolates were genetically grouped into seven clusters, presenting itself as a heterogeneous group.

A remoção de microrganismos patogênicos através da desinfecção é uma necessidade para diminuir a incidência de doenças na população humana relacionadas com poluição fecal. A desinfecção sequencial tem grande potencial na remoção de microrganismos, quando comparada com a desinfecção convencional (um desinfetante). Neste trabalho buscou-se comparar a resistência de microrganismos indicadores de bactérias - E. coli e coliformes totais - e bactérias esporuladas - Clostridium perfringens quanto à desinfecção sequencial empregando cloro seguido de radiação ultravioleta e ozônio seguido de radiação ultravioleta; e à desinfecção convencional utilizando-se os mesmos desinfetantes aplicados individualmente em esgoto sanitário tratado previamente em reator UASB. Os ensaios foram realizados em batelada. As dosagens de cloro aplicadas foram de 10, 20 e 30 mg\'CL IND.2\'/L; de ozônio foram de 5,6; 11 e 16,5 mg\'O IND.3\'/L, ambos para os tempos de contato 10, 20 e 30 minutos. Na desinfecção sequencial com cloro foram aplicadas as doses de 1, 5 e 10 Wh/\'M POT.3\' de radiação UV; com ozônio, as doses de radiação foram variadas de 0,5 a 10 Wh/\'M POT.3\'. Na desinfecção sequencial de cloro-UV, foram removidos 2,5 e 5,2 log de C. perfringens e coliformes totais, respectivamente contra 1,5 e 4,2 log na desinfecção convencional para os mesmos microrganismos. Na desinfecção com cloro, a ordem decrescente de resistência foi: C. perfringens > coliformes totais > E. coli. Para ozônio seguido de UV, C. perfringens apresentou maior resistência e, em alguns ensaios, E. coli apresentou-se mais resistente que coliformes totais. A ação do ozônio mostrou-se notável para a melhoria da qualidade do esgoto tratado avaliada pela diminuição das concentrações de sólidos suspensos totais, sólidos totais, absorbância em comprimento de onda de 254 nm e da DQO, diferentemente do esgoto clorado no qual ocorreu o aumento nos valores destas variáveis físico-químicas. A ação da radiação ultravioleta foi potencializada quando aplicada sequencialmente ao cloro e ao ozônio. / The removal of pathogenic microorganisms through disinfection is a necessity to decrease the incidence of diseases in the human population related to fecal pollution. Sequential disinfection has a great potential on the removal of microorganisms when compared to conventional disinfection (single disinfectant). This work compared the resistance of indicators microorganisms of bacteria - E. coli and total coliforms - and spore-forming bacteria - Clostridium perfringens concerning the sequential and conventional disinfections. The sequential disinfection initially employed chlorine followed by ultraviolet radiation and ozone followed by ultraviolet radiation. The conventional disinfection utilized the same disinfectants individually applied in sanitary sewage previously treated in UASB reactor. The tests were made in batch. The chlorine dosages applied were 10, 20 and 30 mg\'CL IND.2\'/L; ozone were 5,6; 11 and 16,5 mg\'O IND.3\'/L, both for the contact times 10, 20 and 30 minutes. In the sequential disinfection with chlorine dosages of 1, 5 and 10 Wh/\'M POT.3\' of UV radiation were applied; with ozone, the dosages of radiation varied from 0,5 to 10 Wh/\'M POT.3\'. In the sequential disinfection with UV-chlorine were inactived 2,5 and 5,2 log of C. perfringens and total coliforms respectively, against 1,5 and 4,2 log in the conventional disinfection for the same microorganisms. In the chlorine disinfection, the decreasing resistance order was: C. perfringens > total coliforms > E. coli. In the ozone disinfection followed by UV, C. perfringens manifested more resistance and, in some assays, E. coli was more resistant than total coliforms. The action of ozone was outstanding for the improvement of treated sewage quality assessed by the decreasing of total suspended solids concentration, total solids absorbance at a wavelength of 254 nm and of DQO, differently from the chlorine sewage in which occurred an increase of the values of the physical-chemical variables. The ultraviolet radiation action was potentiated when sequentially applied to chlorine and to ozone.

Made available in DSpace on 2014-08-20T13:32:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-10-02 / Avian Necrotic Enteritis (NE) is an acute enterotoxaemia caused by Clostridium perfringens A and C. The control of the disease is based on antibiotics added to animal feed. The ban of this practice by consumer markets, considered the biggest challenge to industrial aviculture, demanded the adoption of other alternatives for its control, among others, immunization with recombinant vaccines. The aim of this work was to produce and evaluate C. perfringens recombinant α toxin (rAT) vaccines adjuvanted with either Al(OH)3 (rAT+Al(OH)3 or recombinant B subunit of the heat-labile enterotoxin of Escherichia coli (rLTB) (rAT+rLTB), and a chimeric protein containing the α toxin fused to rLTB (rLTB-AT). The rAT+Al(OH)3 was innocuous and protected mice against a challenge with native α toxin (sT), and it was immunogenic and did not affect productivity parameters in broilers. The rAT+rLTB showed a dose-protection relationship in mice, while rLTB-AT did not protect mice against sT challenge. The rAT could be an alternative for controlling NE. / A Enterite Necrótica Aviar (ENA) é uma enterotoxemia aguda, causada pelos Clostridium perfringens A e C, cujo controle baseia-se na adição de antibióticos na ração. A restrição dessa prática pelo mercado consumidor, que tornou seu controle o maior desafio para o setor avícola, exigiu a adoção de novas estratégias para o controle, entre elas a imunização. Vacinas recombinantes vêm despertando grande interesse entre pesquisadores e empresas do setor. O objetivo deste trabalho foi elaborar vacinas de toxina α recombinante de C. perfringens (rAT) utilizando como adjuvantes Al(OH)3 (rAT+Al(OH)3) e subunidade B recombinante da enterotoxina termolábil de Escherichia coli (rLTB) (rAT+rLTB), e construir e avaliar uma proteína quimérica contendo rAT fusionada a rLTB (rLTB-AT). A rAT+Al(OH)3 foi inócua e protetora contra agressão de toxina α nativa (sT) em camundongos, e imunogênica em frangos de corte, sem afetar a produtividade. A rAT+rLTB demonstrou relação dose-proteção em camundongos, entanto a rLTB-AT não protegeu camundongos contra agressão de sT. A rAT demonstrou ser uma alternativa para controlar a ENA. Palavras-chave: Enterite Necrótica Aviar, Clostridium perfringens A, toxina α recombinante, vacinas.

Abstract not available

Clostridium perfringens type A isolates producing enterotoxin (CPE) are an important cause of food poisoning and non-food-borne human gastrointestinal (GI) diseases, including antibiotic-associated diarrhea (AAD), and spontaneous diarrhea (SD). In enterotoxigenic type A isolates, the cpe gene is found on the chromosome in food poisoning isolates, but is present on a large virulence plasmid in AAD and SD type A isolates. Food poisoning cases typically exhibit shorter duration of infection and less severe GI symptoms than AAD or SD. Since previous epidemiological evidence has linked the newly discovered beta2-toxin (CPB2) to gastroenteritis in pigs, horses, and chickens, we hypothesize that the CPB2 toxin may be an accessory toxin when cpe positive type A isolates cause human AAD or SD. In the current study, the presence and expression of CPE and CPB2 were assessed in 44 C. perfringens type A human fecal isolates associated with GI diseases in the United Kingdom. Polymerase chain reaction (PCR) and restriction fragment length polymorphisim (RFLP) confirmed the presence of the cpe (32%) and cpb2 (39%) genes. Furthermore, pulsed field gel electrophoresis (PFGE) and I-CeuI RFLP PFGE Southern blot analysis was used to show the localization of the cpe and cpb2 genes, as well as to determine that there was no clonal relationship between the isolates. All surveyed cpb2-positive isolates were determined to carry their cpb2 gene on a large plasmid that was estimated to be the similar size of the cpe large plasmid. Finally, CPE and CPB2 Western blotting demonstrated that all cpe-positive isolates expressed CPE and that all cpb2-positive isolates expressed CPB2. This study identified, for the first time, the C. perfringens non-food-borne human GI disease isolates carrying both the cpe and cpb2 genes (18%), and these isolates all actively expressed both CPE and CPB2. It was also shown that, although CPE expression occurs only under sporulation conditions, CPB2 expressed both in vegetative and sporulation conditions. The CPB2 made by two of these cpe /cpb2 - positive isolates was determined to be very (-99%) similar to the deduced amino acid sequence of the biologically-active CPB2 made by the original type C isolate CWC245. Finally, the expression of CPB2 by only type A isolates carrying the cpe gene on a plasmid and not the isolates carrying a chromosomal cpe gene, could possibly explain the increased GI symptoms and disease duration associated with these non-food-borne GI diseases. Collectively, the current results support a significant association between cpb2-positive C. perfringens isolates and non-food-borne GI disease in human. / Graduation date: 2004

Incorporation of full-fat flaxseed, and to a lesser extent, canola seed in diets to produce n-3-enriched products has attracted interest in the poultry industry. However, high amounts of nonstarch polysaccharides (NSP) in oilseeds compromise their nutritive value. The objectives of the current research were to develop enzyme supplements effective in cell wall depolymerization and viscosity reduction, particularly in flaxseed; to evaluate the effects of enzyme addition and feed processing on oil utilization and egg n-3 fatty acid deposition in broiler chickens and laying hens fed oilseed-containing diets; to characterize the NSP hydrolysis products and to investigate the effects of diet type and enzyme addition on growth performance and the incidence of necrotic enteritis (NE) in broiler chickens challenged with Clostridium perfringens. Results showed that diets containing high levels of flaxseed reduced egg production and shell quality in laying hens, and impaired final body weight and feed conversion ratio (FCR) in broiler chickens. Reducing flaxseed particle size via grinding did not improve the growth performance of broiler chickens, whereas diet pelleting showed more pronounced and beneficial effects in improving the nutritive value of flaxseed, particularly when intact seeds were used. Multicarbohydrase supplementation resulted in a significant depolymerization of cell wall polysaccharides in soybean, canola and flaxseed meals, which was followed by the production of water-soluble NSP hydrolysis products, and the reduction of flax mucilage viscosity in vitro was also evident. Enzyme addition to flaxseed-containing diets improved FCR of broiler chickens and egg production performance of laying hens, and facilitated egg n-3 fatty acid deposition. The C. perfringens challenge caused intestinal NE lesions and increased the mortality of broiler chickens with the highest NE mortality and intestinal C. perfringens counts observed in those fed flaxseed-containing diets. Enzyme supplementation to diets containing high levels of water-soluble NSP (wheat/barley- or wheat/barley/flaxseed-based) facilitated post-disease compensatory growth in pathogen challenged birds. This was accompanied by a numerical reduction of intestinal C. perfringens by 1.4 log10 cfu/g in birds fed the flaxseed-containing diets. Such findings indicated that enzyme addition may be used as a nutritional strategy to reduce the risk of NE development in broiler chickens.

Incorporation of full-fat flaxseed, and to a lesser extent, canola seed in diets to produce n-3-enriched products has attracted interest in the poultry industry. However, high amounts of nonstarch polysaccharides (NSP) in oilseeds compromise their nutritive value. The objectives of the current research were to develop enzyme supplements effective in cell wall depolymerization and viscosity reduction, particularly in flaxseed; to evaluate the effects of enzyme addition and feed processing on oil utilization and egg n-3 fatty acid deposition in broiler chickens and laying hens fed oilseed-containing diets; to characterize the NSP hydrolysis products and to investigate the effects of diet type and enzyme addition on growth performance and the incidence of necrotic enteritis (NE) in broiler chickens challenged with Clostridium perfringens. Results showed that diets containing high levels of flaxseed reduced egg production and shell quality in laying hens, and impaired final body weight and feed conversion ratio (FCR) in broiler chickens. Reducing flaxseed particle size via grinding did not improve the growth performance of broiler chickens, whereas diet pelleting showed more pronounced and beneficial effects in improving the nutritive value of flaxseed, particularly when intact seeds were used. Multicarbohydrase supplementation resulted in a significant depolymerization of cell wall polysaccharides in soybean, canola and flaxseed meals, which was followed by the production of water-soluble NSP hydrolysis products, and the reduction of flax mucilage viscosity in vitro was also evident. Enzyme addition to flaxseed-containing diets improved FCR of broiler chickens and egg production performance of laying hens, and facilitated egg n-3 fatty acid deposition. The C. perfringens challenge caused intestinal NE lesions and increased the mortality of broiler chickens with the highest NE mortality and intestinal C. perfringens counts observed in those fed flaxseed-containing diets. Enzyme supplementation to diets containing high levels of water-soluble NSP (wheat/barley- or wheat/barley/flaxseed-based) facilitated post-disease compensatory growth in pathogen challenged birds. This was accompanied by a numerical reduction of intestinal C. perfringens by 1.4 log10 cfu/g in birds fed the flaxseed-containing diets. Such findings indicated that enzyme addition may be used as a nutritional strategy to reduce the risk of NE development in broiler chickens.

Das MMS stellt eine lebensbedrohliche Erkrankung großwüchsiger Hunde dar. Die ansteigenden Inzidenzzahlen der letzten Jahre, die ungeklärte Ätiologie sowie die hohe Mortalitätsrate rechtfertigen weitere Untersuchungen zu diesem Krankheitskomplex. Daher wurden in der vorliegenden Arbeit Mageninhalte (30 Tiere mit MMS / 13 Kontrolltiere) und Blutproben (102 Tiere mit MMS / 116 Kontrolltiere) mikrobiologisch, biochemisch (kurz-kettige Fettsäuren, Amylase, Lipase, Laktat, pH) und immunologisch (BotNt, CRP, Gesamt-immunglobuline, IgA-, IgG- und IgM-Spiegel ausgewählter mikrobieller Antigene) untersucht. Die Untersuchung des Mageninhaltes der Tiere mit MMS erbrachte signifikant gesteigerte Nachweishäufigkeiten für Hefen, hier v.a. von Rhodotorula mucilaginosa. Des Weiteren konnten in der Gruppe der Tiere mit MMS signifikant gesteigerte pH-Werte sowie signifikant erhöhte Mengen an Acetat, Butyrat und Gesamtfettsäuren im Vergleich zu den Kontrolltieren festgestellt werden. Die gesteigerte bakterielle Fermentation, die ursächlich mit der Entstehung des MMS zusammenhängen dürfte, ist vermutlich auf ein gesteigertes Vorkommen gasbildender Kokken zurückzuführen. Ein Einfluss von Clostridium perfringens an der Entstehung des MMS lässt sich anhand der ermittelten bakteriologischen Ergebnisse nicht belegen. BotNt haben nach den in der vorliegenden Arbeit gewonnenen Daten keinen Einfluss auf die Entstehung des MMS. Serologisch konnten in der Gruppe der Tiere mit MMS signifikant erhöhte CRP-Spiegel gemessen werden. Das CRP ließ aber keine prognostische Aussage über die Überlebenschancen der Tiere zu. Der erhöhte Gesamtimmunglobulin A-Spiegel, bei signifikant erniedrigtem Gesamtimmunglobulin G-Titer weist auf eine vermehrte Auseinandersetzung mit Antigenen hin, die über die geschädigten Schleimhäute eindringen können. Während der Serumlaktatspiegel eine prognostische Aussage über das Outcome der Tiere erlaubt, erscheint eine Bestimmung der Amylase und Lipase beim MMS nicht sinnvoll. Über einen Fragebogen konnte ein gesteigertes Risiko mit zunehmendem Alter, steigender Futtermenge pro Mahlzeit und niedriger Fütterungsfrequenz ermittelt werden. Eine gesteigerte Freßgeschwindigkeit übt einen tendenziellen Einfluss auf die Ausbildung des MMS aus. Keinen Einfluss dahingegen haben das Geschlecht und der Charakter der Tiere, bestehende gastrointestinale Störungen sowie vorangegangene Antibiotikumgaben.