Three yeast antagonists (two strains of Cryptococcus laurentii and one of Candida sake) from orange trees reduced incidence of green mold by 80 to 95% when tested in wounded orange fruits inoculated with Penicillium digitatum and incubated at 7ºC for 30 days. The yeasts inhibited conidial germination of the pathogen, but did not kill the spores. Effectiveness of the three yeasts as antagonists was associated in part with their ability to rapidly colonize wound sites, despite low nutrient availability. Observations suggested that production of extracellular matrix by the yeasts may have facilitated rapid wound colonization. Germination of P. digitatum conidia was significantly inhibited when the pathogen and antagonists were in direct physical contact in a culture suspension. The results supported the view that competition for nutrients is also a mode of action of yeasts against P. digitatum.

No presente estudo avaliou-se o potencial de aplicação da bacteriocina 2a produzida pelo Lactobacillus sake 2a em combinação com outras substâncias antimicrobianas na inibição de cepas de Salmonella isoladas de linguiça: S. Derby SD2, S. Enteritidis SE3, S. Hadar SH4, S. Panama SP5 e S. Typhimurium ST6. A partir de cultura de L. sake 2a em caldo MRS, obteve-se por extração ácida e concentração em liofilizador, um extrato protéico de bacteriocina de cerca de 500 UA/ml. Verificou-se que esse extrato protéico de bacteriocina 2a tem atividade contra Listeria monocytogenes Scott A Cmr Emr nos meios de cultura BHI, MRS e TSB com 0,1% de glicose, independente de temperatura e atmosfera de incubação. O extrato protéico de bacteriocina 2a foi utilizado só e em combinação com EDTA, ácido cítrico, ácido lático ou lisozima sobre "pool" de cepas de Salmonella. Todos os antimicrobianos testados apresentaram efeito inibidor contra Salmonella, no entanto, quando combinados à bacteriocina 2a esse efeito foi mais acentuado. Entre os tratamentos realizados, o que apresentou efeito mais potente na eliminação ou inibição de Salmonella foi a combinação bacteriocina 2a mais ácido lático 0,1%. Bacteriocina 2a também se mostrou mais eficiente que a nisina (usada como padrão) em combinações com lisozima e EDTA. O estudo permitiu verificar que existe um bom potencial de uso da bacteriocina 2a ou do L. sake 2a bac+ em alimentos em associação com os antimicrobianos ácido lático, ácido cítrico, EDTA ou lisozima. Entretanto, penas a combinação de antimicrobianos não é suficiente para eliminar ou inibir a multiplicação de Salmonella. As condições de temperatura, concentração e forma de aplicação dos tratamentos combinados podem variar e são pontos importantes quando se visa à ação direta sobre as células do patógeno ou quando este está associado a um substrato. / The objective of this study was to analyse the potential of application of bacteriocin 2a produced by Lactobacillus sake 2a in combination with other antimicrobials substances on inhibition of strains of Salmonella isolated from raw Brazilian sausages (lingüiça): S. Derby SD2, S. Enteritidis SE3, S. Hadar SH4, S. Panama SP5 and S. Typhimurium ST6. Culture of L. sake 2a grown at 30ºC in MRS broth was used to obtain a cell-free supernatant after centrifugation. This cell-free supernatant was submitted to acid extraction method for bacteriocin and water reduction by liofilization process. After this, a final fraction was obtained and denominated proteic extract of bacteriocin 2a and had a final concentration of approximately 500 A.U/mL. Proteic extract of bacteriocin 2a obtained from cultures of L. sake 2a grown in broths: BHI, MRS, and TSB with 0.1% of glucose showed bactericidal effect against Listeria monocytogenes Scott A Cmr Emr, in any condition of temperature and atmosphere. The proteic extract of bacteriocin 2a was used alone and in combination with EDTA, citric acid, lactic acid or lyzozyme on pool of strains of Salmonella. All antimicrobials tested showed inhibitory effect against Salmonella, but in combination to bacteriocin 2a this effect was more efficient on inhibition or elimination of Salmonella. Bacteriocin 2a showed be more efficient than nisin when in association with lyzozyme and EDTA. This study was able to verify that exist a great potential of application of bacteriocin 2a and/or L. sake 2a bac+, in foods in combination with the antimicrobials: lactic acid, citric acid, EDTA or lyzozyme. However, the simple combination of these antimicrobials is not sufficient to eliminate or inhibit the growth of Salmonella. Temperature conditions, concentration of antimicrobials and form of application of treatments can change and they are very important points; mainly when the aim of the study is to evaluate the direct action of antimicrobials on cells of pathogen or action of these substances on pathogen into a specific substrate.

No presente estudo avaliou-se o potencial de aplicação da bacteriocina 2a produzida pelo Lactobacillus sake 2a em combinação com outras substâncias antimicrobianas na inibição de cepas de Salmonella isoladas de linguiça: S. Derby SD2, S. Enteritidis SE3, S. Hadar SH4, S. Panama SP5 e S. Typhimurium ST6. A partir de cultura de L. sake 2a em caldo MRS, obteve-se por extração ácida e concentração em liofilizador, um extrato protéico de bacteriocina de cerca de 500 UA/ml. Verificou-se que esse extrato protéico de bacteriocina 2a tem atividade contra Listeria monocytogenes Scott A Cmr Emr nos meios de cultura BHI, MRS e TSB com 0,1% de glicose, independente de temperatura e atmosfera de incubação. O extrato protéico de bacteriocina 2a foi utilizado só e em combinação com EDTA, ácido cítrico, ácido lático ou lisozima sobre "pool" de cepas de Salmonella. Todos os antimicrobianos testados apresentaram efeito inibidor contra Salmonella, no entanto, quando combinados à bacteriocina 2a esse efeito foi mais acentuado. Entre os tratamentos realizados, o que apresentou efeito mais potente na eliminação ou inibição de Salmonella foi a combinação bacteriocina 2a mais ácido lático 0,1%. Bacteriocina 2a também se mostrou mais eficiente que a nisina (usada como padrão) em combinações com lisozima e EDTA. O estudo permitiu verificar que existe um bom potencial de uso da bacteriocina 2a ou do L. sake 2a bac+ em alimentos em associação com os antimicrobianos ácido lático, ácido cítrico, EDTA ou lisozima. Entretanto, penas a combinação de antimicrobianos não é suficiente para eliminar ou inibir a multiplicação de Salmonella. As condições de temperatura, concentração e forma de aplicação dos tratamentos combinados podem variar e são pontos importantes quando se visa à ação direta sobre as células do patógeno ou quando este está associado a um substrato. / The objective of this study was to analyse the potential of application of bacteriocin 2a produced by Lactobacillus sake 2a in combination with other antimicrobials substances on inhibition of strains of Salmonella isolated from raw Brazilian sausages (lingüiça): S. Derby SD2, S. Enteritidis SE3, S. Hadar SH4, S. Panama SP5 and S. Typhimurium ST6. Culture of L. sake 2a grown at 30ºC in MRS broth was used to obtain a cell-free supernatant after centrifugation. This cell-free supernatant was submitted to acid extraction method for bacteriocin and water reduction by liofilization process. After this, a final fraction was obtained and denominated proteic extract of bacteriocin 2a and had a final concentration of approximately 500 A.U/mL. Proteic extract of bacteriocin 2a obtained from cultures of L. sake 2a grown in broths: BHI, MRS, and TSB with 0.1% of glucose showed bactericidal effect against Listeria monocytogenes Scott A Cmr Emr, in any condition of temperature and atmosphere. The proteic extract of bacteriocin 2a was used alone and in combination with EDTA, citric acid, lactic acid or lyzozyme on pool of strains of Salmonella. All antimicrobials tested showed inhibitory effect against Salmonella, but in combination to bacteriocin 2a this effect was more efficient on inhibition or elimination of Salmonella. Bacteriocin 2a showed be more efficient than nisin when in association with lyzozyme and EDTA. This study was able to verify that exist a great potential of application of bacteriocin 2a and/or L. sake 2a bac+, in foods in combination with the antimicrobials: lactic acid, citric acid, EDTA or lyzozyme. However, the simple combination of these antimicrobials is not sufficient to eliminate or inhibit the growth of Salmonella. Temperature conditions, concentration of antimicrobials and form of application of treatments can change and they are very important points; mainly when the aim of the study is to evaluate the direct action of antimicrobials on cells of pathogen or action of these substances on pathogen into a specific substrate.

Thesis (Ph. D.)--Rutgers University, 2009. / "Graduate Program in Art History." Includes bibliographical references (p. 344-367).

During alcoholic fermentation Saccharomyces cerevisiae metabolizes L-arginine to ornithine and urea. S. cerevisiae can metabolize urea through the action of urea amidolyase, encoded by the DUR1,2 gene; however, DUR1,2 is subject to nitrogen catabolite repression (NCR) in the presence of high quality nitrogen sources during fermentation. Being cytotoxic at high concentrations, urea is exported into wine where it spontaneously reacts with ethanol, and forms the carcinogen ethyl carbamate (EC). Urea degrading yeast strains were created by integrating a linear cassette containing the DUR1,2 gene under the control of the S. cerevisiae PGK1 promoter and terminator signals into the URA3 locus of the Sake yeast strains K7 and K9. The ‘self-cloned’ strains K7EC- and K9EC- produced Sake wine with 68% less EC. The Sake strains K7EC- and K9EC- did not efficiently reduce EC in Chardonnay wine due to the evolutionary adaptation of said strains to the unique nutrients of rice mash; therefore, the functionality of engineered yeasts must be tested in their niche environments as to correctly characterize new strains. S. cerevisiae possesses an NCR controlled high affinity urea permease (DUR3). Urea importing yeast strains were created by integrating a linear cassette containing the DUR3 gene under the control of the PGK1 promoter and terminator signals into the TRP1 locus of the yeast strains K7 (Sake) and 522 (wine). In Chardonnay wine, the urea importing strains K7D3 and 522D3 reduced EC by 7% and 81%, respectively; reduction by these strains was equal to reduction by the urea degrading strains K7EC- and 522EC-. In Sake wine, the urea degrading strains K7EC- and 522EC- reduced EC by 87% and 84% respectively, while the urea importing strains K7D3 and 522D3 were significantly less capable of reducing EC (15% and 12% respectively). In Chardonnay and Sake wine, engineered strains that constitutively co-expressed DUR1,2 and DUR3 did not reduce EC more effectively than strains in which either gene was expressed solely. Uptake of 14C-urea under non-inducing conditions was enhanced in urea importing strains; parental strains failed to incorporate any 14C-urea thus confirming the functionality of the urea permease derived from the integrated DUR3 cassette.

During alcoholic fermentation Saccharomyces cerevisiae metabolizes L-arginine to ornithine and urea. S. cerevisiae can metabolize urea through the action of urea amidolyase, encoded by the DUR1,2 gene; however, DUR1,2 is subject to nitrogen catabolite repression (NCR) in the presence of high quality nitrogen sources during fermentation. Being cytotoxic at high concentrations, urea is exported into wine where it spontaneously reacts with ethanol, and forms the carcinogen ethyl carbamate (EC). Urea degrading yeast strains were created by integrating a linear cassette containing the DUR1,2 gene under the control of the S. cerevisiae PGK1 promoter and terminator signals into the URA3 locus of the Sake yeast strains K7 and K9. The ‘self-cloned’ strains K7EC- and K9EC- produced Sake wine with 68% less EC. The Sake strains K7EC- and K9EC- did not efficiently reduce EC in Chardonnay wine due to the evolutionary adaptation of said strains to the unique nutrients of rice mash; therefore, the functionality of engineered yeasts must be tested in their niche environments as to correctly characterize new strains. S. cerevisiae possesses an NCR controlled high affinity urea permease (DUR3). Urea importing yeast strains were created by integrating a linear cassette containing the DUR3 gene under the control of the PGK1 promoter and terminator signals into the TRP1 locus of the yeast strains K7 (Sake) and 522 (wine). In Chardonnay wine, the urea importing strains K7D3 and 522D3 reduced EC by 7% and 81%, respectively; reduction by these strains was equal to reduction by the urea degrading strains K7EC- and 522EC-. In Sake wine, the urea degrading strains K7EC- and 522EC- reduced EC by 87% and 84% respectively, while the urea importing strains K7D3 and 522D3 were significantly less capable of reducing EC (15% and 12% respectively). In Chardonnay and Sake wine, engineered strains that constitutively co-expressed DUR1,2 and DUR3 did not reduce EC more effectively than strains in which either gene was expressed solely. Uptake of 14C-urea under non-inducing conditions was enhanced in urea importing strains; parental strains failed to incorporate any 14C-urea thus confirming the functionality of the urea permease derived from the integrated DUR3 cassette.

During alcoholic fermentation Saccharomyces cerevisiae metabolizes L-arginine to ornithine and urea. S. cerevisiae can metabolize urea through the action of urea amidolyase, encoded by the DUR1,2 gene; however, DUR1,2 is subject to nitrogen catabolite repression (NCR) in the presence of high quality nitrogen sources during fermentation. Being cytotoxic at high concentrations, urea is exported into wine where it spontaneously reacts with ethanol, and forms the carcinogen ethyl carbamate (EC). Urea degrading yeast strains were created by integrating a linear cassette containing the DUR1,2 gene under the control of the S. cerevisiae PGK1 promoter and terminator signals into the URA3 locus of the Sake yeast strains K7 and K9. The ‘self-cloned’ strains K7EC- and K9EC- produced Sake wine with 68% less EC. The Sake strains K7EC- and K9EC- did not efficiently reduce EC in Chardonnay wine due to the evolutionary adaptation of said strains to the unique nutrients of rice mash; therefore, the functionality of engineered yeasts must be tested in their niche environments as to correctly characterize new strains. S. cerevisiae possesses an NCR controlled high affinity urea permease (DUR3). Urea importing yeast strains were created by integrating a linear cassette containing the DUR3 gene under the control of the PGK1 promoter and terminator signals into the TRP1 locus of the yeast strains K7 (Sake) and 522 (wine). In Chardonnay wine, the urea importing strains K7D3 and 522D3 reduced EC by 7% and 81%, respectively; reduction by these strains was equal to reduction by the urea degrading strains K7EC- and 522EC-. In Sake wine, the urea degrading strains K7EC- and 522EC- reduced EC by 87% and 84% respectively, while the urea importing strains K7D3 and 522D3 were significantly less capable of reducing EC (15% and 12% respectively). In Chardonnay and Sake wine, engineered strains that constitutively co-expressed DUR1,2 and DUR3 did not reduce EC more effectively than strains in which either gene was expressed solely. Uptake of 14C-urea under non-inducing conditions was enhanced in urea importing strains; parental strains failed to incorporate any 14C-urea thus confirming the functionality of the urea permease derived from the integrated DUR3 cassette. / Medicine, Faculty of / Medical Genetics, Department of / Graduate

This study was designed to assess the difference in litigation between two courts: one mandating For Kids' Sake, a psycho-educational seminar for divorcing parents, and the other not so mandating. The level of difficulty of children's adjustment to divorce has been positively correlated with parental hostility. More hostile parents would have more contested cases, interim motions, and relitigations. This research compared final dispositions, interim motions, and relitigation between parents in two courts in Collin County, Texas. The treatment group was from the 219th District Court which mandated all divorcing parents with minor children to attend the For Kids' Sake Seminar and the control group was from the 199th District Court which did not so mandate. Archival data was collected from a computer generated list for the Total group data to assess final dispositions and directly from District Clerk files for the In-Depth group data to assess interim motions and relitigation. The Total group was comprised of 679 research subjects with 330 cases in the treatment group and 349 cases in the control group. The In-Depth group consisted of 182 cases from both courts with 84 cases in the treatment group and 98 cases in the control group. Chi square analysis of the total group revealed significantly more parents in the treatment group who non suited the divorce suit and remained married (p. < .05), a significantly lower number of cases in the treatment group with interim motions (p. < .10), and a significantly lower amount of relitigation in the treatment group (p. < .05). The results showed that the court that mandated For Kids' Sake evidenced a reduction in subsequent litigation which not only benefits the legal system but also hopefully reflects lower parental hostility and higher parental cooperation, thereby benefiting the children of divorce.

A comparative study was performed to evaluate the immunostimulatory effect of yeast and yeast-derived glucan in white prawn Fenneropenaeus indicus (sub-adults of ∼20 gm). Feed with a whole cell biomass of marine yeast Candida sake S165 (CSY) at a concentration of 10% (w/w) and another feed with 0.2% glucan of C. sake S165 (CSG) were used in the study. Fenneropenaeus indicus were fed with these diets for 40 days and subsequently challenged with the white spot syndrome virus (WSSV). Haematological parameters such as the total haemocyte count, phenoloxidase activity, superoxide anion (O2-) level, haemolymph peroxidase level and post-challenge survival against WSSV infection were determined to assess the immune status. In the present experiment, a higher immunity index and post-challenge survival were recorded in shrimps fed with the whole cell yeast diet. The better immunostimulatory performance of the whole cell yeast diet compared with the glucan diet could be attributed to the cellular constituents of yeast including the cell wall glucan, nucleotides, carotenoid pigments and vitamins. Here we observed that whole cell yeast performed better as an immunostimulant than the extracted cell wall glucans. Therefore, the use of yeast biomass in diets, rather than the yeast cell wall extract, glucan, would confer better protection against microbial infection besides reducing the cost of shrimp production.

O efeito combinado de bacteriocina produzida por Lacfobacillus sake 2ª e embalagem em atmosfera modificada sobre o controle de Lisferia monocytogenes F5069r em lingüiça frescal foi avaliado. A cepa L. sake 2a foi co-inoculada com L. monocytogenes F5069r (resistente a cloranfenicol e eritromicina) em lingüiça frescal. As lingüiças foram embaladas com ar, 100% CO2 ou 50%CO2/50%N2 e armazenadas a 6°C. A multiplicação de L. Monocytogenes F5069r e L. sake 2a foi monitorada durante 4 semanas em intervalos de 7 dias. A avaliação sensorial, por meio do teste triangular, foi realizada após 5 e 11 dias, os quais foram estipulados de acordo com a vida-de-prateleira do produto. Após 28 dias de estocagem, a população de L. monocytogenes nas amostras inoculadas com L. sake 2a e embaladas com atmosfera modificada foi 6.4 ciclos logarítmicos menor que no controle sem a bactéria lática e embalado em ar. No entanto, a influência da atmosfera modificada sobre as características sensoriais do produto foram detectáveis após cinco dias de estocagem, independente da adição de L. sake 2a. Ao final da primeira semana, a influência de L. sake 2a sobre L. monocytogenes foi menos importante (redução de 0,4 log, não significante) que a influência da embalagem em atmosfera modificada (redução de 1,4 log, significante). No décimo primeiro dia, nenhuma diferença sensorial foi encontrada entre as amostras com e sem L. sake 2a embaladas em atmosfera modificada. Após 14 dias, a população de L. monocyfogenes nas amostras com L. sake 2ª embaladas em atmosfera modificada foi 3,5 log menor que no controle sem a bactéria lática e embalado em ar. Os resultados sugerem que o uso combinado de atmosfera modificada e bacteriocina produzida por L. sake 2a apresenta um efeito sinergístico sobre o controle de L. monocytogenes F5069r em lingüiça frescal refrigerada. / Lactobacillus sake 2a is a bacteriocinogenic strain isolated from \"lingüiça frescal\", a Brazilian sausage. The combined effect of modified-atmosphere packaging and addition of L. sake 2a on inhibition of L. monocyfogenes in \"lingüiça\" was evaluated. Samples were inoculated with L. monocytogenes and/or L. sake 2a, packed with oxygen-permeable film, 100%CO2 or 50%CO2+50%N2 and stored at 6°C. Microbial counts were performed weekly. Sensorial evaluation (triangle tests with 16 subjects) was performed after 5 and 11 days (shelf-life). After the fourth week, L. monocytogenes population in samples packed with modified atmosphere containing L. sake 2a was 6.4 log lower than in samples without any treatment. However, the influence of the modified atmosphere on the sensorial characteristics of the product was already detectable on the fifth day (a risk of 5%), regardless the addition of L. sake 2a. By the end of the first week, the influence of L. sake 2a on the inhibition of L. Monocytogenes was less important (reduction of 0.4 log, non significant) than the influence of the packaging (reduction of 1.4 log, significant). On the 11th day, no significant sensorial difference was found between the samples with and without L. sake 2a packed with modified atmosphere. By the end of the second week, L.monocytogenes counts in samples packed with modified atmosphere containing L. sake 2a were 3.5 log lower than counts in samples without any treatment. Combination of results suggests that modified atmosphere and L. sake 2a act synergistically on inhibition of L. monocytogenes in \"lingüiça frescal\".