Thesis MSc(Agric)--Stellenbosch University, 1979. / ENGLISH ABSTRACT: no abstract available / AFRIKAANSE OPSOMMING: geen opsomming

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: During fermentation, nitrogenous compounds serve as nutrients for the yeasts, which enable their growth, functioning and maintenance of the yeasts cells. From a winemaking perspective, a certain amount of nitrogen is required for the yeasts in order to avoid sluggish or stuck fermentation. Moreover, nitrogen metabolism leads to the production of aroma compounds such as higher alcohols, fatty acids and esters which contribute positively to overall sensory characteristics of wine. Nitrogen metabolism (uptake of ammonium and amino acids) have been extensively studied in Saccharomyces cerevisiae. Nonetheless, the fairly great variances observed between strains in terms of preference for certain nitrogen sources and metabolism thereof are not so well understood. Additionally, these mechanisms nitrogen metabolism of non- Saccharomyces yeasts are even vaguer and simply assumed to be globally similar to those of S. cerevisiae. This study aimed to investigate the uptake of nitrogen compounds (ammonium and individual amino acids) by selected non-Saccharomyces yeasts (Lachancea thermotolerans IWBT Y1240, Torulaspora delbrueckii Biodiva TD291, Pichia kluyveri FrootZen, Metschnikowia pulcherrima IWBT Y1123 and Metschnikowia pulcherrima Flavia) to assess the impact of fermentation kinetics and the production of aroma compounds during sequential fermentations with S. cerevisiae under different initial YAN concentrations, with 300 mg/L, 150 mg/L and 75 mg/L, respectively). Fermentations were performed in a synthetic grape juice medium with pure and sequential fermentations. The data showed that the assimilation of nitrogen compounds were species specific. For example, L. thermotolerans preferred alpha amino nitrogen above ammonia, where the opposite hold true for T. delbrueckii. Notable differences could also be identified for the uptake of certain single amino acids. Irrespective of the initial YAN concentrations during sequential fermentations, the yeasts only assimilated about half of the initial YAN. The non-Saccharomyces yeasts did not influence fermentation performance during sequential fermentations. However, a low initial YAN (75 mg/L) had a strong influence on the fermentation kinetics and aroma compound production. The higher uptake (compare to S. cerevisiae) of specific single amino acids by non-Saccharomyces yeasts (especially L. thermotolerans), can be tentatively correlated with certain aroma compounds produced at the end of fermentation. The results also revealed that agitation could impact overall fermentation performance and aroma compound production. This study contributes to an improved understanding of how different initial nitrogen concentrations affect growth, fermentation performances and aroma compound production of wine-related yeasts under fermentative conditions. Moreover, the uptake of single amino acids by selected non-Saccharomyces yeasts had also been identified, which is a good starting point to better understand non- Saccharomyces yeasts nitrogen requirements which may be used for the optimization of nitrogen source addition, during alcoholic fermentation, when used in mixed fermentations in order to ensure a complete alcoholic fermentation. To the best of our knowledge, the uptake of single amino acids and YAN consumption by selected non-Saccharomyces yeasts under fermentation conditions tested, have never been studied before. / AFRIKAANSE OPSOMMING: Tydens wynfermentasies dien talle stikstof komponente as voedingstowwe vir wyngis wat hul groei, funksie en onderhoud bevorder. Van `n wynmaak perspektief word daar `n sekere hoeveelheid stikstof benodig deur die wyngis om te verhoed dat slepende of onvolledige fermentasies plaasvind. Verder lei stikstofmetabolisme na die produksie van aroma verbindings, soos hoër alkohole, vlugtige vetsure en esters wat positief bydra tot die sensoriese karaktereienskappe van wyn. Die stikstofmetabolisme (opneem ammonium en aminosure) is deeglik nagevors in die wyngis Saccharomyces cerevisiae, maar die klein variasies waargeneem tussen die gisras in terme van die voorkeur van sekere stikstof komponente is egter nog onduidelik. Daarbenewens is die stikstofmetabolisme nog meer onbekend in nie- Saccharomyces wyngis en word dit oor die algemeen aanvaar dat die werking van die stikstofmetabolisme dieselfde is as in S. cerevisiae. Hierdie studie het gestreef om die opneem van stikstof komponente (ammonium en aminosure) te ondersoek van uitverkiesde nie-Saccharomyes gis (Lachancea thermotolerans IWBT Y1240, Torulaspora delbrueckii Biodiva TD291, Pichia kluyveri FrootZen, Metschnikowia pulcherrima IWBT Y1123 and Metschnikowia pulcherrima Flavia) deur te bepaal wat die impak is op die groei-kinetika en op die produksie van aroma komponente gedurende gemengde kultuur fermentasies met S. cerevisae onder verskillende aanvangs assimileerbare stikstof (300 mg/L, 150 mg/L en 75 mg/L). Fermentasies is in sintetiese druiwemos uitgevoer vir beide enkel en gemengde kultuur fermentasies. Die resultate demonstreer dat die assimilasie van stikstof ras spesifiek was. Byvoorbeeld, L. thermotolerans verkies alfa amino stikstof bo ammonium waar die teenoorgestelde waar is vir T. delbrueckii. Beduidende verskille is ook waargeneem vir die opneem van sekere individuele aminosure. Die wyngis het steeds net die helfte van die assimileerbare stikstof opgeneem gedurende gemengde kultuur fermentasies ongeag die aanvangsstikstof konsentrasies. Die nie-Saccharomyces gis het nie die fermentasie kinetika beïnvloed tydens gemengde kultuur fermentasies nie. Daar was egter ook waargeneem dat `n lae assimileerbare stikstof (75 mg/L) `n negatiewe invloed op die fermentasie kinetika sowel as aroma produksie gehad het. Die hoër opname (vergelyking met S. cerevisiae) van sekere aminosure deur nie-Saccharomyces gis, kan tydelik gekoppel word aan die produksie van spesifieke aroma verbindings aan die einde van fermentasies. Die resultate het ook gewys dat die toepassing van skud `n impak het op die fermentasie kinetika sowel as die produksie van aroma komponente. Die studie dra by om beter te verstaan van hoe verskillende aanvangsstikstof die groei, fermentasie kinetika en aroma produksie beïnvloed onder fermentasie kondisies. Die opneem van sekere aminosure deur nie-Saccharomyces gis word ook beskryf, wat `n goeie beginpunt is om beter te vertaan wat die stikstof vereistes vir die geselekteerde wyngis is, wat gebruik kan word vir die optimisering van stikstofaanvullings, sodat die risiko van probleemfermentasies verlaag sal word. So ver as wat ons kennis strek is die opneem van aminosure en die gebruik van assimileerbare stikstof deur nie-Saccharomyces wyngis onder fermentasie kondisies nog nie ondersoek nie.

Diets were based on steam-flaked milo and contained chopped alfalfa hay in the control diet. At the 65% concentrate level, cottonseed hulls or chopped wheat straw replaced half the alfalfa hay. At the 90% concentrate level, roughage sources were chopped alfalfa hay, cottonseed hulls, or wheat straw. Six growing steers and three mature, rumen-cannulated steers per concentrate level were used in separate Latin square experiments. Total tract digestion coefficients, particulate passage rates, liquid turnover rates, and rumination time were measured in the intact steers. In situ digestion, rumen pH, and rumen dry matter distribution were evaluated in the cannulated steers. Competition between rates of passage and digestion were used to calculate apparent extent of ruminal digestion (AED). At the 65% concentrate level, total tract digestion coefficients for the wheat straw vs alfalfa hay diet were not different, apparently due to increased milo and alfalfa hay neutral detergent fiber (NDF) digestion in the wheat straw diet. Milo and alfalfa hay AED for NDF were higher (P<.10). Dietary AED for DM, NDF, and cell solubles were highest (P<.05) for the alfalfa hay diet. Differences between diets were minimal, indicating roughage source in 90% concentrate diets does not substantially influence milo digestion or passage as it does in 65% concentrate diets.

Total tract digestion coefficients for dry matter (DM) and neutral detergent fiber (NDF), passage rates for grain and roughage components of diets and liquid turnover were measured for 65 and 90% concentrate diets in separate 4 x 4 Latin square experiments using intact growing steers. Kinetics of in situ digestion for DM and NDF in diet ingredients, rumen pH, volume and DM distribution were determined in mature, rumen cannulated steers. Rates of passage and digestion were combined to calculate apparent extent of ruminal digestion (AERD) for diet ingredients. Diets were based on steam flaked milo. The control roughage was chopped alfalfa hay (AH). In 65% concentrate diets, chopped wheat straw (WS), bermudagrass straw (BS) or cottonseed hulls (CSH) replaced 50% of the AH. In 90% concentrate diets, all of the AH was replaced by WS, BS or CSH. Total tract starch digestion was not influenced by source of roughage at either concentrate level. In 65% concentrate diets, total tract digestion of DM and NDF was not adversely affected by substitution of WS for 50% of the AH. Digestion of these fractions was lower (P<.05) for BS and CSH diets, but the severity of depression was greatest (P<.05) for the CSH diet. Reduced digestibility of the CSH diet was attributed to lower digestibility of CSH in comparison with AH. Although WS and BS were also less digestible than AH, their inclusion in the diet improved (P<.05) AERD of DM and NDF from milo and AH. Improved AERD for NDF appeared to be related to the raft-forming properties of WS and BS, and their ability to maintain rumen pH in a more desirable range for NDF digestion. In 90% concentrate diets, effects of roughage source on utilization of milo were minimal. Total tract digestion of DM and NDF was lower (P<.05) for WS, BS and CSH diets than for the AH diet, because of dilution of the more digestible AH by these roughages. Wheat straw had a tendency to moderate ruminal pH and consequently might be more useful in high concentrate diets than roughages such as CSH and BS.

No description available.

Based on ethanol fermentation kinetics and bioreactor engineering theory, a system composed of a continuously stirred tank reactor (CSTR) and three tubular bioreactors in series was established for continuous very high gravity (VHG) ethanol fermentation with Saccharomyces cerevisiae. Sustainable oscillations of residual glucose, ethanol, and biomass characterized by long oscillation periods and large oscillation amplitudes were observed when a VHG medium containing 280 g/L glucose was fed into the CSTR at a dilution rate of 0.027 h???1. Mechanistic analysis indicated that the oscillations are due to ethanol inhibition and the lag response of yeast cells to ethanol inhibition. A high gravity (HG) medium containing 200 g/L glucose and a low gravity (LG) medium containing 120 g/L glucose were fed into the CSTR at the same dilution rate as that for the VHG medium, so that the impact of residual glucose and ethanol concentrations on the oscillations could be studied. The oscillations were not significantly affected when the HG medium was used, and residual glucose decreased significantly, but ethanol maintained at the same level, indicating that residual glucose was not the main factor triggering the oscillations. However, the oscillations disappeared after the LG medium was fed and ethanol concentration decreased to 58.2 g/L. Furthermore, when the LG medium was supplemented with 30 g/L ethanol to achieve the same level of ethanol in the fermentation system as that achieved under the HG condition, the steady state observed for the original LG medium was interrupted, and the oscillations observed under the HG condition occurred. The steady state was gradually restored after the original LG medium replaced the modified one. These experimental results confirmed that ethanol, whether produced by yeast cells during fermentation or externally added into a fermentation system, can trigger oscillations once its concentration approaches to a criterion. The impact of dilution rate on oscillations was also studied. It was found that oscillations occurred at certain dilution rate ranges for the two yeast strains. Since ethanol production is tightly coupled with yeast cell growth, it was speculated that the impact of the dilution rate on the oscillations is due to the synchronization of the mother and daughter cell growth rhythms. The difference in the oscillation profiles exhibited by the two yeast strains is due to their difference in ethanol tolerance. For more practical conditions, the behavior of continuous ethanol fermentation was studied using a self-flocculating industrial yeast strain and corn flour hydrolysate medium in a simulated tanks-in-series fermentation system. Amplified oscillations observed at the dilution rate of 0.12 h???1 were postulated to be due to the synchronization of the two yeast cell populations generated by the continuous inoculation from the seed tank upstream of the fermentation system, which was partly validated by oscillation attenuation after the seed tank was removed from the fermentation system. The two populations consisted of the newly inoculated yeast cells and the yeast cells already adapted to the fermentation environment. Oscillations increased residual sugar at the end of the fermentation, and correspondingly, decreased the ethanol yield, indicating the need for attenuation strategies. When the tubular bioreactors were packed with ????? Intalox ceramic saddles, not only was their ethanol fermentation performance improved, but effective oscillation attenuation was also achieved. The oscillation attenuation was postulated to be due to the alleviation of backmixing in the packed tubular bioreactors as well as the yeast cell immobilization role of the packing. The residence time distribution analysis indicated that the mixing performance of the packed tubular bioreactors was close to a CSTR model for both residual glucose and ethanol, and the assumed backmixing alleviation could not be achieved. The impact of yeast cell immobilization was further studied using several different packing materials. Improvement in ethanol fermentation performance as well as oscillation attenuation was achieved for the wood chips, as well as the Intalox ceramic saddles, but not for the porous polyurethane particles, nor the steel Raschig rings. Analysis for the immobilized yeast cells indicated that high viability was the mechanistic reason for the improvement of the ethanol fermentation performance as well as the attenuation of the oscillations. A dynamic model was developed by incorporating the lag response of yeast cells to ethanol inhibition into the pseudo-steady state kinetic model, and dynamic simulation was performed, with good results. This not only provides a basis for developing process intervention strategies to minimize oscillations, but also theoretically support the mechanistic hypothesis for the oscillations.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Master of Science / Food Science and Industry / Fadi M. Aramouni / Gluten-free products are becoming more prevalent in the market today, however there are a few types of products that have “hidden” gluten and people will not realize until after consumption. Products like soy sauce and beer are sources of gluten that people don’t know about. Soy sauce contains wheat as a main ingredient so replacing it with a gluten-free flour such as sorghum may produce a product similar to wheat-based soy sauce. Sorghum was used in this experiment since it is a grain grown in the mid-western region of the United States and a growing food ingredient in the global market. Sorghum can come in many different varieties and colors so we used different varieties in this study. Four treatments were done using three different sorghum flours (black, white, and waxy sorghum flour) and a wheat flour for a control. Cooked soybeans were mashed in a kitchenaid mixer and the treatment flour was added to make a dough. That dough was formed into a log and cut into slices. The slices were then staked with wet paper towels to mold. After 13 days of molding, the slices that were made were dried, placed in a salt solution, and fermented for 100 days. The solution was mixed with a spatula for 30 seconds to homogenize the mixture every 2-3 days and samples were taken every 10 days to test for pH, salinity, and color. Once the 100 days were complete and the pH of each treatment did not drop for consecutive testing periods, the liquid was removed from the solids and pasteurized. The pH curve did show that a fermentation process did occur, however there was no control over what microorganism could grow. Consumer testing was not performed since all test sauces were deemed unacceptable at the initial screening.

Doctor of Philosophy / Department of Human Nutrition / Mark D. Haub / Little is known about the impact of dietary fiber (DF) on children. Current recommendations are based on extrapolations from adult studies. Research is needed to provide science based evidence to determine how DF impacts the gut of children. Two studies were conducted to investigate the interactions of DF in the child large intestine. In the first study, the dose response of DF on breath hydrogen, methane, and total hydrogen content was investigated relative to Institute of Medicine (IOM) recommended intakes in free-living preschool children. Only four of the 18 participants were able to comply with the treatment protocol. Although, no significant differences were noted in breath measures of fermentation across fiber consumption levels, there was a numerical pattern for increasing levels of DF to evolved increased gas production in the four compliers. In free-living individuals measures of acute fiber fermentation through breath was not sensitive enough over 6 hours to distinguish a difference in fermentative rate. Children, parents, and child care centers found this approach apparently acceptable. In study two, the impact of DF (10 g) fed over three weeks in children and their parents on metabolic markers of fermentation were evaluated. The body was able to adapt to 10 g/day DF consumption as bloating (p < 0.05) and flatulence (p = 0.06) decreased each week of the study. Fecal propionic acid was significantly increased over three weeks of DF supplementation. There was also an interaction (p=0.05) between time and age for butyric acid. Dietary fiber supplementation (10 g/day) over three weeks via a commercially available extruded cereal was well-tolerated by the participants, with no disturbances in bowel habit in children or adults. Alternatively, there were no improved bowel habit measures with increased DF consumption. This study provides evidence that this tolerable dose of DF supplementation over three weeks had similar impacts in free-living children and adults. However, the presence of increased butyric acid only in children may be reflective of different production or absorptive capacities between children and adults. Although not presented here, the bacterial ecological analysis may shed further insight into the interactions occurring in the large intestine. These are the first studies to my knowledge to have investigated these outcomes in young children. In addition to the gut health outcomes, this research provided a framework into the apparent feasibility of studying children in a gentle, non-invasive, and cost-effective manner.