Thesis (Ph. D.)--Ohio State University, 2003. / Title from first page of PDF file. Document formatted into pages; contains xx, 323 p. Includes abstract and vita. Advisor: Shang-Tian Yang, Dept. of Chemical Engineering. Includes bibliographical references (p. 263-288).

Master of Science / Department of Biological and Agricultural Engineering / Wenqiao Yuan / Many of the current industrial fermentation processes cannot use pentose as the carbon source. However, complete substrate utilization of sugars in lignocellulose is one of the prerequisites to render economic development of biofuels or bioproducts from biomass. In this study we proposed a new process for DHA production from renewable carbon sources by first using anaerobic bacteria, Clostridium tyrobutyricum to convert pentose into organic acids with butyric acid as the main product, and then using the organic acids to feed microalgae, Crypthecodinium cohnii to produce DHA. The effect of glucose and xylose on the yield of butyric acid produced by C. tyrobutyricum was investigated, separately. Cell growth of C. tyrobutyricum increased with increasing initial glucose or xylose concentration, but was not affected significantly when the concentration was above 55g/l for glucose or 35g/l for xylose. Butyric acid yield increased as the initial sugar concentration increased in both xylose and glucose, but the conversion rate from xylose or glucose to butyric acid decreased as the sugar concentration increased. The xylose to glucose ratio in their mixture did not significantly affect cell growth or butyric acid yield. The effect of arabinose on the yield of butyric acid produced by C. tyrobutyricum was also studied. As for butyric acid production, compared with glucose or xylose, the arabinose was in a low efficiency, with butyric acid output of 2.25g/l in 10g/l arabinose and a long lag period of about 3-4 d. However, a low concentration of arabinose could be used as a nutritional supplement to improve the fermentability of a mixture of xylose and glucose. The conversion rate of sugar to butyric acid increased as the supplement arabinose increased. In order to obtain low cost xylose, corncobs were hydrolyzed and this xylose-rich product was used to culture C. tyrobutyricum. The results showed that at end of the 9 d fermentation, the concentration of butyric acid from corncob hydrolysate reached 10.56 g/l, and the mimic medium reached 11.3 g/l. This suggests that corncob hydrolysate can be used as a carbon source for butyric acid production by C. tyrobutyricum, although some inhibitory effects were found on cell growth with corncob hydrolysate. The effect of butyric acid, lactic acid and acetic acid on the yield of DHA produced by C. cohnii was also investigated, separately. The DHA yield was highly related to both biomass and DHA content in the cell, whereas lower growth rate could bring higher DHA content. The best concentration for DHA yield seemed to be 1.2g/l in three single organic acid media. In two organic acids mixture media, acetic acid tended to be beneficial for biomass accumulation, regardless whether butyric acid or lactic acid was mixed with acetic acid, the OD could reach 1.3 or above. When butyric acid was mixed with lactic acid, the highest DHA yield was achieved, due to increased DHA content from mutual influence between butyric acid and lactic acid.

<p>One of the most serious and economically important defects caused by clostridia in milk products is the late blowing of semi-hard cheeses.</p><p>Clostridia occur naturally in soil and can contaminate milk through crops contaminated by dung and soil followed by a less successful silage process, that give them opportunity to grow unaerobically. When anaerobic conditions occur, such as storage of semi-hard cheese, they ferment lactic acid to butyric acid and the gases CO₂ and H₂.</p><p>At the fusion of Arla and MD Foods, a series of changes were conducted on the MPN method for lactic acid fermentation for clostridia in milk. These changes resulted in an increased accuracy due to an increased number of test tubes and the change of media from MRCM to BBB, Bryant & Burkey Broth, that was thought to be more selective for <em>Cl. tyrobutyricum</em>, the organism mostly found in hard cheese. When the number of dairy farmers that were given quality reduction fines increased, the new method was suspected and a validation was conducted.</p><p>The validation included inoculation of different clostridia and bacillus strains into BBB substrate and enzymatic testing of positive samples with Rapid ID 32A. The inoculation result showed that almost all tested different clostridia strains could grow in BBB substrate.</p><p>Test on BBB positive tubes with Rapid ID 32A resulted in 95% clostridia of which 70% was <em>Cl. tyrobutyricum</em>. These results correlated well with earlier studies on MRCM substrate and the increase in quality reduction fines probably depended on the larger number of test tubes used in the new method rather than the change of substrate.</p> / <p>Clostridier eller främst <em>Cl. tyrobutyricum</em> är den art som i de flesta fall orsakar feljästa ellersönderjästa ostar, vilket är ett välkänt problem inom mejeriindustrin. Clostridiesporer finns naturligt i jord och hamnar i mjölkråvaran via gröda som kontaminerats med gödsel och jord och som tillsammans med en mindre lyckad ensileringsprocess gör att clostridierna växer till. När anaeroba förhållanden uppstår, såsom vid lagring av hårdost, förjäser clostridierna laktat och smörsyra varvid vätgas och koldioxid bildas.</p><p>I samband med fusionen av Arla/MD Foods, genomfördes metodförändringar för MPN-metoden för laktatjäsande clostridier i mjölk, som används inom mjölkbedömningen. Dessa förändringar innebar en skärpning i noggrannheten genom att man ökade antal rör, samt ett substratbyte från MRCM till BBB, Bryant & Burkey Broth, som ansågs mer gynnsamt för <em>Cl. tyrobutyricum</em>.</p><p>Då antalet mjölkproducenter som fick kvalitetsavdrag ökade, riktades misstankar mot den nya metoden och en validering genomfördes.</p><p>Resultatet av valideringen, som innebar ympning av renkulturer, utodling av positiva rör med efterföljande typning, visade att de flesta av de tillsatta stammarna av clostridier hade förmågan att ge ett positivt utslag vid tillsats av renkulturer. Utodling och typning av positiva rör visade att ca 70 % av de positiva utslagen innehöll <em>Cl. tyrobutyricum</em> och hela 95 % någon clostridieart. Dessa resultat korrelerar med tidigare studier på MRCM-substrat och ökningen av positiva utslag berodde troligtvis på det ökade antalet rör som den nya metoden innebar.</p>

One of the most serious and economically important defects caused by clostridia in milk products is the late blowing of semi-hard cheeses. Clostridia occur naturally in soil and can contaminate milk through crops contaminated by dung and soil followed by a less successful silage process, that give them opportunity to grow unaerobically. When anaerobic conditions occur, such as storage of semi-hard cheese, they ferment lactic acid to butyric acid and the gases CO2 and H2. At the fusion of Arla and MD Foods, a series of changes were conducted on the MPN method for lactic acid fermentation for clostridia in milk. These changes resulted in an increased accuracy due to an increased number of test tubes and the change of media from MRCM to BBB, Bryant &amp; Burkey Broth, that was thought to be more selective for Cl. tyrobutyricum, the organism mostly found in hard cheese. When the number of dairy farmers that were given quality reduction fines increased, the new method was suspected and a validation was conducted. The validation included inoculation of different clostridia and bacillus strains into BBB substrate and enzymatic testing of positive samples with Rapid ID 32A. The inoculation result showed that almost all tested different clostridia strains could grow in BBB substrate. Test on BBB positive tubes with Rapid ID 32A resulted in 95% clostridia of which 70% was Cl. tyrobutyricum. These results correlated well with earlier studies on MRCM substrate and the increase in quality reduction fines probably depended on the larger number of test tubes used in the new method rather than the change of substrate. / Clostridier eller främst Cl. tyrobutyricum är den art som i de flesta fall orsakar feljästa ellersönderjästa ostar, vilket är ett välkänt problem inom mejeriindustrin. Clostridiesporer finns naturligt i jord och hamnar i mjölkråvaran via gröda som kontaminerats med gödsel och jord och som tillsammans med en mindre lyckad ensileringsprocess gör att clostridierna växer till. När anaeroba förhållanden uppstår, såsom vid lagring av hårdost, förjäser clostridierna laktat och smörsyra varvid vätgas och koldioxid bildas. I samband med fusionen av Arla/MD Foods, genomfördes metodförändringar för MPN-metoden för laktatjäsande clostridier i mjölk, som används inom mjölkbedömningen. Dessa förändringar innebar en skärpning i noggrannheten genom att man ökade antal rör, samt ett substratbyte från MRCM till BBB, Bryant &amp; Burkey Broth, som ansågs mer gynnsamt för Cl. tyrobutyricum. Då antalet mjölkproducenter som fick kvalitetsavdrag ökade, riktades misstankar mot den nya metoden och en validering genomfördes. Resultatet av valideringen, som innebar ympning av renkulturer, utodling av positiva rör med efterföljande typning, visade att de flesta av de tillsatta stammarna av clostridier hade förmågan att ge ett positivt utslag vid tillsats av renkulturer. Utodling och typning av positiva rör visade att ca 70 % av de positiva utslagen innehöll Cl. tyrobutyricum och hela 95 % någon clostridieart. Dessa resultat korrelerar med tidigare studier på MRCM-substrat och ökningen av positiva utslag berodde troligtvis på det ökade antalet rör som den nya metoden innebar.

Thesis (doctoral)--Swedish University of Agricultural Sciences, 2000. / Includes bibliographical references.

No description available.

Made available in DSpace on 2015-03-26T13:52:02Z (GMT). No. of bitstreams: 1 texto completo.pdf: 443818 bytes, checksum: 511a55303b3b59b278ccd2fab938970a (MD5) Previous issue date: 2006-04-28 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The mango (Mangifera indica Linn.) is one of the most important tropical fruits. It is very appreciated due to its flavor and nutritional characteristics. The fruits are easily spoiled because of intrinsic factors, which makes difficult its commercialization in natura. An alternative to reduce the losts is the conservation of the mango as pulp, juice and nectar. The thermal treatment is the most used process to conservate these products. However, the complete inativation of microorganisms during the thermal processing adopted by most industries of fruit juices and fruit pulps is limited by the presence of spore formers and termorresistents microorganisms. This work aimed to isolate and to identificate spoilage bacteria from thermally processed mango pulp, and to characterize the fermentative activity of these microorganisms. Samples of 25 lots of spoiled mango pulp from two industrial units processing mango pulp were evaluated in plate count agar, added with 20 % of steril mango pulp. A number of 104 bacteria were isolated and were characterized as Gram-positive rods, spore formers and mostly strict anaerobic, indicating that belonged to the genus Clostridium. Four isolated named LMA 45, LMA 63, LMA 72 and LMA 104, provenient of different lots of mango pulp from two industrial units, were identified by biochemical tests and by the cell membrane fatty acids profile as Clostridium tyrobutyricum. These four isolates were selected for use in the experiments that followed. The isolates were cultivated in BHI broth within a pH range of 3.0 to 7.0 and temperatures between 7 °C to 50 °C. C. tyrobutyricum grew in the pH range of 4.0 to 7.0, except for isolate LMA 104, that did not grow in pH 4.0 up to 48 hours of incubation. None of the isolates was able to grow in pH 3.5. The specific growth rate (&#956;) was higher in pH values between 6.5 and 7.0 and varied from 0.18 to 0.31 h-1. The growth was observed in a temperature range from 10 °C to 40 °C, with an optimum between 35 °C and 40 °C. The fermentative activity of C. tyrobutyricum LMA 45, LMA 63, LMA 72 and LMA 104 was evaluated in mango pulp with pH varying from 3.0 to 5.0. Gas production was detected in pH of 4.0, 4.5 and 5.0 before 50 hours of the incubation at 30 ºC, except for isolate LMA 104 which produced gas only after 100 hours of incubation. The other main products that resulted from mango pulp fermentation in pH values between 4.0 and 4.5 were acetate and butyrate, which was determined by high performance liquid chromatography (HPLC). The consumption of sucrose, glucose and frutose during mango pulp fermentation in pH 4.0 and 4.5 was also determined using HPLC. The C. tyrobutyricum isolates evaluated could grow at high sugar concentrations, as they fermented the mango pulp containing 729,2 mM of sugars, but did not use all the substrate. The growth of the isolates was inhibited in mango pulp at pH 3.5. This results suggest the adoption of the acidification strategy by the industries, wich generally process mango pulp with pH 4.0. This strategy might ensure the conservation of the fruit product, without changes in the thermal treatment. / A manga (Mangifera indica Linn.) é uma das mais importantes frutas tropicais, muito apreciada pelo sabor, aroma e por suas propriedades nutricionais. Os frutos são perecíveis em razão dos fatores intrínsecos, dificultando a sua comercialização in natura. Uma alternativa para reduzir as perdas é a conservação da manga na forma de polpa, suco ou néctar. O tratamento térmico é o processo mais empregado para a conservação desses produtos. Entretanto, a inativação completa de microrganismos durante o processamento térmico normalmente adotado pelas indústrias processadoras de sucos e polpas é limitada pela presença de organismos formadores de esporos e de termorresistentes. Este trabalho teve como objetivos isolar e identificar as bactérias deterioradoras de polpa de manga tratada termicamente e caracterizar a atividade fermentativa desses microrganismos. Amostras de 25 lotes de polpa de manga deteriorada provenientes de duas unidades industriais processadoras de polpa de manga foram analisadas em ágar padrão para contagem, enriquecido com 20 % de polpa de manga esterilizada. Um total de 104 isolados foi obtido e essas bactérias foram caracterizadas como bastonetes, Gram-positivas, formadoras de esporos sendo a grande maioria constituída de anaeróbias estritas, do gênero Clostridium. Destes, os isolados LMA 45, LMA 63, LMA 72 e LMA 104 obtidos de lotes diferentes de polpa de manga provenientes de duas unidades industriais, foram identificados por teste bioquímicos e pelo perfil de ácidos graxos de membrana como Clostridium tyrobutyricum e selecionados para a continuidade deste estudo. Os isolados foram cultivados em caldo BHI com pH variando de 3,0 a 7,0 e em temperatura entre 7 °C e 50 °C. O crescimento de C. tyrobutyricum ocorreu em valores de pH de 4,0 a 7,0 com exceção do isolado LMA 104 que não apresentou crescimento em pH 4,0 em até 48 horas de incubação. Não houve crescimento em pH 3,5. A velocidade específica de crescimento (&#956;) foi maior em pH entre 6,5 e 7,0 e alcançou valores entre 0,18 e 0,31 h-1. O crescimento foi observado em temperaturas de 10 °C a 40 °C, com ótimo entre 35 °C e 40 °C. A atividade fermentativa de C. tyrobutyricum LMA 45, LMA 63, LMA 72 e LMA 104 foi avaliada em polpa de manga com pH variando de 3,0 a 5,0. A produção de gás foi detectada em valores de pH de 4,0, 4,5 e 5,0 em menos de 50 horas de incubação a 30 ºC, com exceção do isolado LMA 104 cuja produção de gás foi detectada após 100 horas de incubação. Além de gás, os principais produtos da fermentação da polpa de manga com pH entre 4,0 e 4,5 foram acetato e butirato, determinados por cromatografia líquida de alta eficiência (HPLC). O consumo de sacarose, glicose e frutose durante a fermentação de polpa de manga com pH 4,0 e 4,5 foi determinado utilizando-se HPLC. Os isolados de C. tyrobutyricum avaliados apresentaram capacidade de crescer em altas concentrações de açúcar, pois fermentaram a polpa de manga com 729,2 mM de açúcares e não utilizaram todo o substrato durante a fermentação. A inibição do crescimento dos isolados em polpa de manga com pH 3,5 permite sugerir a adoção da estratégia de acidificação para garantir a conservação do produto pela indústria, que geralmente processa esse produto com pH 4,0.

Made available in DSpace on 2015-03-26T13:52:03Z (GMT). No. of bitstreams: 1 texto completo.pdf: 406557 bytes, checksum: f8e8d62d3ae14f56df99a0e2bc645da0 (MD5) Previous issue date: 2006-04-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Bacteriocins from lactic acid bacteria have been suggested as an alternative to traditional food preservation methods, such as heat treatment, that interfere with natural characteristics of the food. Nisin is the bacteriocin that has been most used in foods. Recently, a new bacteriocin, bovicin HC5, produced by Streptococcus bovis HC5, was characterized. Previous work indicated that this bacteriocin has activity against Listeria monocytogenes and Clostridium sporogenes. In this study, the activity of this bacteriocin was tested against Bacillus cereus, Bacillus thuringiensis and Clostridium tyrobutyricum isolated form spoiled mango pulp. The addition of bovicin HC5 (40 to 160 AU/mL) into BHI media resulted in reduced specific growth rate and maximal optical densities of B. cereus, B. thuringiensis and C. tyrobutyricum. Concentrations of 40 and 80 AU/mL increased lag phase duration for at least 10 h. When 160 AU/mL was used, growth was not observed even after 144 h. The effect of bovicin HC5 against vegetative cells inoculated into mango pulp was bactericidal and more pronounced at acidic conditions. After 24 h of incubation with the bacteriocin, the viable cell number was bellow the detection level. Similar results were obtained when nisin was used. When C. tyrobutyricum was inoculated into mango pulp with 100 AU/mL of bovicin HC5, gas production was not observed for at least 10 days of incubation. Bovicin HC5 reduced spore germination of B. cereus and B. thuringiensis inoculated into mango pulp, and the number of non-germinated spores after 122 h of incubation was at least 100-fold greater than control treatments. Bovicin HC5 did not affect the thermal resistance of B. cereus and B. thuringiensis spores. However, if added to the mango pulp, bovicin HC5 could remain stable during the heat treatment, and reduce spore germination of these microorganisms. Cultures of B. cereus, B. thuringiensis and C. tyrobutyricum that were transferred successively in the presence of subletal doses of bovicin HC5 did not become resistant. Considering the results obtained in this study and the fact that bovicin HC5 was stable in culture supernatants and in mango pulp, it seems that this bacteriocin could be useful to prevent the spoilage of mango pulp by B. cereus, B. thuringiensis and C. tyrobutyricum. / Bacteriocinas de bactérias do ácido láctico têm sido propostas como alternativa aos métodos tradicionais de conservação de alimentos, como tratamento térmico, que interferem nas características naturais do alimento. A nisina é a bacteriocina que tem sido mais utilizada em alimentos. Recentemente, uma nova bacteriocina, bovicina HC5 produzida por Streptococcus bovis HC5, foi caracterizada e apresentou efetividade contra Listeria monocytogenes e Clostridium sporogenes. Neste estudo, a atividade desta bacteriocina foi testada contra Bacillus cereus, Bacillus thuringiensis e linhagens de Clostridium tyrobutyricum isolados da polpa de manga deteriorada. A adição de 40 a 160 UA/mL de bovicina HC5 em caldo BHI resultou na diminuição da velocidade específica de crescimento e da DO máxima atingida por B. cereus, B. thuringiensis e C. tyrobutyricum. Concentrações de 40 e 80 UA/mL de bovicina HC5 resultaram no aumento da fase lag dos isolados em pelo menos 10 h e quando a concentração utilizada foi de 160 UA/mL, o crescimento não foi observado por, pelo menos, 144 h. Concentração de 100 UA/mL de bovicina HC5 foi bactericida para células vegetativas inoculadas em polpa de manga e este efeito foi mais pronunciado em condições acídicas. Após 24 horas de incubação na presença da bacteriocina, o número de células viáveis das bactérias avaliadas ficou abaixo do limite de detecção. Resultados semelhantes foram obtidos com a nisina. Quando as linhagens de C. tyrobutyricum foram inoculadas em polpa de manga contendo 100 UA/mL de bovicina HC5, a produção de gás não foi observada por até 10 dias de incubação. Bovicina HC5 reduziu a germinação de esporos de B. cereus e B. thuringiensis e o número de esporos no estado dormente após 122 h de incubação foi, pelo menos, 100 vezes maior do que o observado no tratamento controle, sem bacteriocina. Esta bacteriocina não teve efeito na resistência térmica de esporos de B. cereus e B. thuringiensis. Entretanto, uma vez que a bovicina HC5 apresenta resistência a altas temperaturas (121 ºC/20 min), ela pode permanecer estável na polpa de manga após o tratamento térmico e reduzir a germinação dos esporos sobreviventes. A transferência dos isolados por, aproximadamente, 40 gerações na presença de 20 UA/mL de bovicina HC5 não causou adaptação dos microrganismos deterioradores. Esta bacteriocina permaneceu estável após incubação em mistura com o sobrenadante das culturas e com a polpa de manga. Considerando os resultados obtidos neste trabalho, a bovicina HC5 parece ser útil para reduzir a deterioração de polpa de manga causada por B. cereus, B. thuringiensis e C. tyrobutyricum.

Although the etiology and pathogenesis of inflammatory bowel disease (IBD) is not fully understood, it is generally accepted that the inflammation results from aberrant immune responses to antigens of gut microbiota in genetically susceptible individuals (Sartor et al., 2006). Alteration in intestinal microbiota has been found in IBD patients with increased abundance of certain bacteria and decreased abundance of others. Due to the complexity of the disease, multifaceted interactions between genetic factors, host immune response, gut microbiota and environment factors need to be taken into account. In this thesis, the pathogenesis of IBD was first reviewed in respect with the four factors mentioned above. Then we concentrated on the interaction between IBD-associated bacteria and mucosal immune system. We investigated the ability of mucosal-associated bacteria (MAB) from IBD patients to induce spontaneous colitis in germ-free (GF) mice and the impact of those bacteria on the development of dextran sulfate sodium (DSS)-colitis. Together with the analysis of the composition of gut microbiota of MAB colonized mice, we demonstrated the potential deleterious microbes were able to increase the susceptibility to DSS-colitis once they found a suitable niche. We revealed the mechanism of an E.coli strain...