Thesis (M.S. in engineering)--Washington State University, December 2006. / Includes bibliographical references.

Based on the initial exploratory results of single-phase (acidogenesis and methanogenesis takes place in one vessel) whey biomethanation studies, a two-phase (acidogenesis and methanogenesis takes place in two separated serial vessels) biomethanation process was found to be more suitable for dealing with the current whey utilisation and/or disposal problem. Acidogenesis was found to be less understood in comparison to methanogenesis and therefore acidogenesis became the central problem of this thesis. Given that 90% of the five-day biochemical oxygen demand in whey is due to lactose, continuous culture (Chemostat) experiments were undertaken to examine the general mechanism of lactose acidogenesis by a mixed undefined culture using ¹⁴C-labeled tracers. Also the influence of whey protein (mainly β-lactoglobulin) on the general fermentation scheme was addressed. Experimental factors included a pH range of 4.0 to 6.5, a mesophilic temperature of 35°C and a dilution rate (D) range of 0.05 to 0.65 h⁻¹. At a fixed pH level, the observed variability in the main acidogenic end products (acetate, propionate, butyrate and lactate) with respect to D were found to be a consequence of the systematic separation of the various microbial groups involved in acidogenesis. Batch incubation of a [¹⁴C(U)]-lactate tracer with chemostat effluent samples and preparative separation of the end products followed by a liquid scintillation assay of the location of the radio activity demonstrated that a microbial population lactate to other end products and hence the observed increase in lactate concentrations at high D values. Further use of [¹⁴C(U)]-butyrate and [¹⁴C(2)]-propionate revealed the predominant carbon flow routes from pyruvate to the various end products. A qualitative lactose acidogenic fermentation model was proposed, in which lactose is converted to pyruvate via the Embden-Meyerhof-Parnas pathway. Pyruvate in a parallel reaction is then converted to lactate and butyrate. In the presence of hydrogen reducing methanogens lactate is converted to acetate in a very fast reaction and not propionate as previously believed-. The implications of these findings with regard to optimising the acidogenic phase reactor are discussed. Acidogenic fermentation of protein together with lactose did not affect the carbon flow scheme. In the D range of 0.05 to 0.15 h⁻¹ low pH (pH < 5.0) was found to favour the butyrate route at the expense of the lactate route and at high pH (pH > 5.5) the lactate route was favoured at the expense of the butyrate route, the pH region of 5.0 to 5.5 being the transition range. In order to describe the microbial growth, the Monod chemostat model was chosen among the various alternatives, because of its simplicity and its physico-chemical basis. The estimated model parameters are reported. / Applied Science, Faculty of / Chemical and Biological Engineering, Department of / Graduate

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Resistance exercise training (RET) is a well-known stimulus for muscle protein synthesis. Protein supplementation, in conjunction with RET, has been shown to yield greater accretion of lean body mass than RET alone. Few studies have compared two multi-ingredient, isonitrogenous supplements of differing quality protein. Therefore, the purpose of the current study was to determine whether there was an augmented effect of a high-quality whey protein multi-ingredient nutritional supplement on hypertrophy in young adults following a RET program. We hypothesized that the multi-ingredient supplement would induce hypertrophy to a greater extent than the control supplement in young adults. Twenty-six (13 male, 13 female) healthy young adults (22 ± 2 years [mean ± SD]) were randomly assigned to either the multi-ingredient nutritional supplement (SUPP, n=12: 20g whey protein, 2g leucine, 2.5g creatine monohydrate, 300mg calcium citrate, 1000IU vitamin D) or control beverage (CON, n=12: 20g collagen protein, 1.4g alanine, 0.6g glycine) groups, ingesting their respective supplements twice daily. Measurements were obtained prior to and after a 10-week linear RET program. Dual-energy X-ray absorptiometry (DXA), ultrasound, one-rep maximum (1RM), and biopsies from the vastus lateralis muscle were performed. A 2-way ANOVA (time by supplement) revealed significantly larger increases in lean body mass (LBM), as assessed via DXA, from the active supplement compared to the control (SUPP: +4.1 ± 1.3kg CON: +2.8 ± 1.7kg, p=0.004). We conclude that the consumption of a multi-ingredient nutritional supplement increased lean body mass to a greater extent than to that observed in the CON group in healthy young adults. / Thesis / Master of Science in Kinesiology / Resistance exercise training (RET) is known to augment muscle size, a process known as hypertrophy. Several factors may affect hypertrophy, such as supplementation with protein and amino acids, increasing an individual’s potential to maximize muscle hypertrophy beyond RET-induced gains. However, little is known regarding the effectiveness of multi-ingredient supplements and the synergistic effects they may exhibit on hypertrophy and strength gains. The present thesis shows the effect of a specific multi-ingredient supplement that contained: whey protein, creatine, calcium, vitamin D, and leucine, compared to a low-quality collagen-based supplement, on measures of hypertrophy and strength in young adults. Interestingly, the active supplement (SUPP) induced greater gains in lean body mass (LBM), type II muscle fibre CSA, and bicep CSA and thickness compared to the control (CON), but not strength. These findings provide insight into a novel formulation of ingredients on exercise-induced increases in hypertrophy in young adults.

The traditional filter paper assay for saccharifying cellulase originally described by M. Mandels et al (1976) has been modified to make possible low activity determinations of Trichoderma cellulases. The enzymatic activity appears to decline during a prolonged incubation period if no precautions have been taken. By means of adding bovine serum albumin and potassium chloride as protein stabilizers and sodium azide as an antimicrobial agent filter paper activities in the range from 0.02 to 0.37 (IUPAC assay, 1987) can be estimated by extending the incubation time up to 20 hours. Filter paper activity values obtained by this method may be compared to those obtained by the IUPAC assay by using a conversion factor from 1.4 to 1.7. Acid whey and sweet whey permeate have been investigated as media for growth and metabolite production by Trichoderma reesei QM 9414 using shake flask cultures and spore inocula. In the case of acid whey the mycelial growth after 2 weeks is 13 mg dry weight /ml substrate. The specific growth rate is 0.29/day. The fungus appears to metabolize the whey protein the first 2 weeks. The alkalinity of acid whey rises continuously over a three week period up to a pH of 8.5. In the case of whey permeate the maximal mycelial weight gain is 4.4 mg/ml which appears after 8 days. A rise in net soluble protein level comes after 3-5 days and reaches a maximum value of 0.23 mg/ml after 2 weeks. The pH of whey permeate rises continuously to 7.5 after 3 days and then slowly declines. The net production of cellulases is low on both media. Dilution 1:6 of the acid whey, supplementation with ammonium sulfate and pHadjustments did not enhance the production of cellulases. Acid whey supports a significant growth and sweet whey permeate shows potential for extracellular protein production. A literature review surveys the composition and uses of acid whey, environmental aspects of whey wastes, the fungus Trichoderma reesei, the mode of action of the Trichoderma reesei cellulase system and the structure of cellulose in cotton and wood. / Graduation date: 1994

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