Increasing charges by the private utilities for the treatment of industrial waste water are making on-site effluent treatment more attractive. On-site anaerobic digestion is increasingly being used by food processing factories as a cost effective solution to waste liquid waste disposal. Discharge of treated effluent to sewer or water course requires compliance to a maximum admissible concentration (MAC) value, therefore, there is a need for careful control of on-site waste water treatment. This research investigates the treatment of effluent from instant coffee production. This results in a liquid waste that contains recalcitrant and toxic compounds formed during the roasting process. This waste varies in strength and composition according to the different processes that are performed in the manufacture of instant coffee. Anaerobic filters are particularly attractive for wastes containing recalcitrant or inhibitory compounds requiring a long sludge age. Therefore, this study was aimed at firstly investigating the treatability of coffee waste, using anaerobic filters; and secondly monitoring and control of the digestion process in order to maintain a constant effluent quality.

Blood-feeding and digestion are crucial for the tick reproduction because they provide nutrients for anabolic processes such as vitellogenesis, molting and eggs production. Digestion in ticks is mediated by a network of cystein and aspartic peptidases. Therefore, tick digestive peptidases could be a relevant anti-tick vaccine candidates.

Au sevrage, la séparation de la mère, le changement d’environnement et la transition d’un régime lacté à un régime solide et complexe déstabilisent les fonctions de digestion et de défense du porcelet. Le sevrage entraîne l’apparition de diarrhées nécessitant souvent l’utilisation d’antibiotiques pour contrôler les infections digestives. L’émergence de souches bactériennes antibiorésistantes est devenue un enjeu de santé publique majeur. Développer des méthodes alternatives aux antibiotiques pour promouvoir la santé du porcelet sevré est une nécessité. L’objectif de la thèse est d’évaluer le potentiel de deux microalgues –la Spiruline et la Chlorelle– pour promouvoir la santé digestive du porcelet au sevrage.Deux essais ont été effectués chez des porcelets sevrés dans des conditions d’hygiène contrastées –standard ou dégradées– recevant ou non 1% de Spiruline ou de Chlorelle dans l’aliment. En conditions d’hygiène standard, la supplémentation en Spiruline ou en Chlorelle a amélioré la digestibilité des nutriments et la morphologie intestinale sans impact sur la croissance du porcelet sevré. En conditions d’hygiène dégradée, la supplémentation en Spiruline ou en Chlorelle a altéré les performances de croissance sans affecter sensiblement les indicateurs de la santé digestive. Dans un troisième essai, une administration orale en microalgues en amont et en aval du sevrage a été testée afin d’évaluer la capacité de la Spiruline ou de la Chlorelle à limiter la déstabilisation précoce du système digestif au sevrage. L’administration orale de Spiruline a favorisé la croissance e / Weaning is a critical step in pig farms. Separation from the sow, change in the environment and dietary transition from liquid milk to a solid diet induces a destabilization of digestive and defense functions in weaned pig. Weaning leads to digestive disorders and growth alteration that often require antibiotic use to alleviate pathogen invasion and mortality. Bacterial resistance to antibiotics has become a major concern for public health. Thus, it has become necessary to find new strategies to promote digestive health in weaned pigs. The aim of this thesis is to evaluate the potential of two microalgae – Spirulina and Chlorella – as new functional ingredients to promote digestive health in piglets at weaningTwo trials were conducted in pigs weaned in contrasted sanitary conditions –good or poor–and that received 1% of Spirulina or Chlorella in starter diet. In good sanitary conditions, dietary supplementation with Spirulina and Chlorella both enhanced nutrient digestibility and intestinal morphology but did not affect growth performance. In poor sanitary conditions, both Spirulina and Chlorella supplementation altered growth performance with slight effect on indicators of digestive health. In a third trial, oral administration of the two microalgae before and after weaning was assayed to evaluate the ability of Spirulina and Chlorella to limit the early intestinal changes caused by weaning. Spirulina administration before and after weaning enhanced growth in suckling piglets and reduced risk of diarrhoea outcome in weaned pigs. Potential mechanisms of action of Spirulin

The gastric morphology of the white-tailed rat M. albicaudatus - was described in detail. The bilocular hemiglandular stomach consists of a papillated corpus, non-glandular PGP and glandular antrum. The antrum contains cardiac, fundic and PJ loric glands (suggesting limited glandular reduction during gastric evolution) while the FCE and PGP are lined with orthokeratin. The corpal papillae, which increase surface area for microbial attachment, have undergone a different type of keratinisation called physiological hyperkeratosis. Streptococci, Lactobacilli and unidentified anaerobic bacilli (which colonise papillary microhabitats) are autochthonous in the stomach of M. albicaudatus but P. vulgaris and Ps. flourescens are probably autochthonous. Early gastric development is innate but the rapid development of PB into papillae between 15 and 17 days suggests the presence of allogenic growth stimuli: possibly mechanical abrasion by solid food, low chalone concentration in the papillary basal cells and the influence of the APB. Stimulation by VFA presence, however, is unlikely due to the low concentration of these acids in the stomach (Summary, p. 225)

The evaluation of potential alternative protein sources for the formulation of least-cost optimal diets to satisfy the nutrient requirements of South African abalone (Haliotis midae) has been hampered by the absence of a suitable, practical, replicable and reliable digestibility technique. A suitable lowcost faeces collection technique was developed in this study to obtain suitable quantities of excreta for analysis from this species. Acid-insoluble ash was identified as a reliable, replicable and safe internal marker in comparison to chromic oxide and crude fibre for use in nutrient digestibility studies with H. midae. This was validated by the consistency and repeatability of the results and by comparison to total collection of faeces. The traditional substitution method used in digestibility studies with fish to evaluate protein digestibility of feed ingredients was found to be unsuitable for H. midae. Apparent protein digestibility values exceeding 100 % derived through this method could be attributed to associative effects between feed ingredients, differential diet and faecal nutrient leaching, and mathematical artifacts in calculations when using substitution versus single protein diets. An ingredient particle size of less than 450 μm in comparison to particle size classes of above 450 μm was shown to enhance nutrient (dry matter, organic matter, protein, fat) digestibility and minimise dry matter leaching from diets. The dietary inclusion level of both pre-gelatinised maize starch and a-cellulose did not influence (P > 0.05) apparent nutrient (protein, fat, fiber, starch) digestibility. Using the above digestibility protocol amino acid availability of all plants ingredients currently used in the South African animal industry was evaluated for H. midae. Soybean meal (96.86 %) and lupins (96.51 %) presented the highest apparent mean amino acid availability of all plant protein ingredients evaluated with H. midae. Canola meal (94.21 %), faba beans (92.87 %) sunflower meal (92.77 %), peanut meal (87.39 %) and cottonseed meal (85.15 %) presented higher apparent mean amino acid availability values than fish meal (82.75 %). Apparent protein digestibility was highly correlated (r = 0.99) with mean apparent amino acid availability, while true amino acid availability was 1.88 % units higher than apparent amino acid availability for all ingredients tested. Predicted apparent protein digestibility in compound diets was within 1.1-6.5 % of determined values. Calcium phosphate mono dibasic presented the lowest (P < 0.05) dietary phosphorus leaching (51.51 % maximum) and highest apparent phosphorus digestibility (66.27 %) in comparison to other inorganic phosphorus sources. Based on the method of direct experimentation to determine the optimal dietary protein level using graded levels of dietary protein 28.1-35.9 % dietary protein from good quality sources is recommended for maximum growth of juvenile H. midae. This study provides a scientifically sound research tool including a faecal collection technique, suitable marker and assay technique that could be use in further studies to improve least-cost diet formulation for H. midae. Future nutritional studies in H. midae should primarily concentrate on reducing dietary nutrient leaching and improving the intake of nutrients in order to properly evaluate responses of this species to different dietary regimes.

The potential to reduce feed formulation costs by the replacement of existing proteinsources in artificial feeds for Haliotis midae was assessed. A comparison between the efficacy of the direct method and chromic oxide marker techniques in determining apparent protein digestibility coefficients, revealed that while the former was not a suitable methodology for use with this species, the latter produced replicable and reliable results. It was established that the chromic oxide marker is inert, is not absorbed by the abalone, does not interfere with the digestive processes and moves through the intestine at the same rate to the protein. However, as this method was time consuming and expensive to implement, a multienzyme pH-stat in vitro protein digestibility technique using a three enzyme system was employed to rapidly assess the protein quality of 34 protein sources. The efficacy of the technique was established by correlating the in vitro digestibility estimates with in vivo digestibility coefficients obtained using the chromic oxide marker technique. The highest degree of in vivo predictability was attained when protein sources were separated according to origin, and significant correlations between either animal (r²=0.89, P<0.004) or plant (r²=0.79, P<0.04) protein sources were found. The effect of animal size on the qualitative protein requirements of two size classes of H.midae was assessed by feeding 12 isonitrogenous and isoenergetic single protein diets to juvenile and young adult animals (10-20 and 40-50mm shell length). The criteria for protein source selection were their bioavailability as determined using the pH-stat in vitro digestibility technique, and their cost and availability within South Africa. The protein sources identified for the trial comprised four fishmeals, casein, spirulina, abalone viscera silage, brewery waste, torula yeast, carcass, sunflower and cotton meals. The results indicated that in terms of growth and feed efficiency, the fishmeals and spirulina were the most suitable candidates as primary protein sources in formulated feeds for H.midae, and with the exceptions of the carcass meal and brewery waste, the remaining protein sources demonstrated promise as partial primary protein source replacements. The smaller size class of abalone displayed significantly reduced growth, feed and protein efficiency than their larger counterparts. In terms of feed conversion and growth response, two-way analysis of variance revealed significant interactions between protein source and animal size, suggesting that qualitative differences exist between the dietary protein requirements of the juvenile and young adult abalone. The commercial implication of thisfinding was discussed. An evaluation was undertaken to determine the effects of the partial and total replacement of dietary fishmeal with selected plant protein sources on growth and nutritional indices of juvenile H.midae. A commercial “Abfeed” formulation in which 100% of the protein component comprised LT-fishmeal was employed as a control. Fifteen isonitrogenous experimental diets were formulated in which the LT fishmeal was substituted at either 10, 15, 30, 50, 75 or 100% with either spirulina, semolina, ground maize, torula yeast, soya, sunflower or corn gluten meals or combinations thereof. No significant differences were found in the growth rates between the control diet and those diets in which 30% of the fishmeal component had been replaced by either soya, sunflower meal, or torula yeast. In addition, 50% of the fishmeal component could be substituted with either soya meal or spirulina without affecting growth. Replacement of either 75 or 100% of the fishmeal had a significant negative affect on growth. Pearson product moment correlations between dietary lysine levels and either growth rates or protein efficiency ratios revealed positive correlations (r=0.77, P=0.0005; r=0.52, P=0.04 respectively) suggesting that lysine may have been the first limiting amino acid in these diets. Carcass analysis revealed that dietary protein source had no significant effect on body composition. An assessment of the dietary arginine requirement of juvenile H.midae using whole proteins to supply graded levels of dietary arginine did not promote a growth response. It was concluded that arginine is probably not the first limiting amino acid in formulated feeds for H.midae. An assessment of the dietary lysine requirement of juvenile H.midae using two micro-encapsulation techniques (gelatin/acacia colloid or cellulose acetate phthalate) was not successful. The efficacy of the encapsulation techniques were established and an assessment of the degree of lysine supplementation undertaken. Failure of the crystalline lysine enriched diets to promote growth suggests that the prospects for using crystalline amino acids in essential amino acid requirement studies in H.midae is low. The results of the present study indicate that the prospects for replacing fishmeal with cheaper alternative protein sources in artificial feeds for H.midae is promising. Furthermore, while the technical difficulties such as the determination of the essential amino acid requirements of the abalone precluded the application of “least cost” programming, the prospects for its future application are promising.

No description available.

No description available.

Part I of this thesis describes the digestion of bakers' yeast s-RNA at 0° and at room temperature in the presence of various concentrations of magnesium ions and at several different s-RNA : enzyme ratios (w/w). Digestion of s-RNA with pancreatic ribonuclease (ratio, 2000:1, (w/w) ) in the presence of 0. 2 M magnesium and at 0°, followed by chromatographic analysis, indicated that about 75% of the ultraviolet absorbing material was of medium to high molecular weight. Analysis of the nucleotide composition of the fraction containing the intermediate size oligonucleotides showed that fraction to be enriched in the odd bases ψ, T, 1-methylG, N, N-dimethylG and several other unidentified nucleotides. This result was incompatible with the 'hairpin' models for s-RNA, proposed by McCully and Cantoni (1962). The presence of spermine in the digestion mixture clearly masked the catalytic action of pancreatic ribonuclease, albeit to a lesser extent than magnesium ions, contrary to a report by Thomas and Hubst (1963) in their study of the affect of spermine on the catalytic activity of E. coli ribonuclease toward E. coli ribosomal RNA. The studies described in Part II of this thesis are concerned with the fractionation of mixed s-RNA from yeast. Part II-A describes a new chromatographic material consisting of a bifunctional mercuri-dioxane derivative attached through a thiol group to the cellulose matrix. A previous report (Eldjarn and Jellum, 1963) had shown that HS-proteins could be fractionated on an organomercurial-cellulose (Material I) derived from aminoethyl-cellulose. The studies reported, here suggest that ¹⁴C-cysteinyl-s-RNA cannot be retained on Material I because the salt concentration required to overcome its ion-exchange properties preclude any interaction between the mercury of Material I and the sulfhydryl group of ¹⁴C-cysteinyl-s-RNA. The present study describes how this disadvantage was overcome by preparing an organomercurial-cellulose devoid of ion-exchange properties (Material II). Preliminary studies showed that Material II can retain the radioactivity associated with ¹⁴C-cysteinyl-s-RNA. These studies suggested that Material II might fractionate nucleic acids according to their base composition. Part II-B describes some of the attempts to fractionate yeast s-RNA by column partition chromatography. In general, resolution of acceptor activities of s-RNA, as well as the recovery of biological activity, was poor. Part II-C describes some of the author's contributions to the studies carried out in this laboratory on the fractionation of yeast s-RNA on another new chromatographic material, benzoylated-DEAE-cellulose (Material III). Certain variables (such as, magnesium ion concentration and pH), which can be manipulated during rechromatography of s-RNA on Material III, are discussed in light of the chromatographic behaviour of glycine and other acceptor RNA's. The forces responsible for this fractionation are also discussed. Part II-D describes a chemical procedure for the isolation of glycine s-RNA which, when combined with the chromatographic procedures described in Part II-C, afforded glycine-specific s-RNA, in high purity. Observations on the effect of magnesium ion during enzymatic synthesis of glycyl-s-RNA are discussed. The advantages and possible consequences of using purified aminoacyl-s-RNA synthetase enzymes for the preparation of the aminoacyl-s-RNA's are also discussed. The need to prepare large quantities of aminoacyl-s-RNA synthetases derives from their indispensable role in the development of chemical methods for purifying amino acid-specific s-RNA's (see Part II-D). Grinding of yeast cells with glass beads, which is the usual method employed for disrupting yeast cells, was found to be inadequate. Lysis of yeast cells with toluene at 37° was found to be a partial solution to this problem. Although previous studies showed that most of the synthetase enzymes were destroyed by this treatment (von Tigerstrom and Tener, 1967), investigations described in Part III of this thesis, show that most of the synthetase activities can be preserved by controlling the pH of the toluene-yeast mixture. However, there were still several synthetase activities missing in the cell-free extracts prepared by the warm toluene method. Preparation of large quantities of yeast cell-free extract containing all of the aminoacyl-s-RNA synthetases was finally achieved by treatment of the yeast cells with toluene containing excess dry ice followed by an incubation period at 3-5°. Partial purification of the aminoacyl-s-RNA synthetases was achieved by chromatographing the cell-free preparation on hydroxyl-apatite. The presence of 40% glycerol in the eluting buffers was found to be essential for the preservation of most of the synthetase activities. When the cell-free preparations were chromatographed on hydroxy-apatite, most of the protein, but only a few of the synthetases were eluted by a linear gradient of phosphate buffer to 0.20 M. Most of the synthetases were eluted only when the column was washed with ammonium sulfate. The significance of this observation is discussed. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

The anaerobic digestion of cheese whey was studied in an upfiow anaerobic sludge blanket reactor for its start-up characteristics, the effects of various process parameters, the effect of sulfate addition and the determination of optimal operating conditions. Start-up of an UASB reactor treating cheese whey was extremely difficult due to its tendency to acidify. Various start-up strategies were tested to facilitate start-up and to ensure stable operation. Among the operating parameters, sludge loading rate was the most critical for proper start-up of the UASB reactor. The initial sludge loading rate during start-up period should not exceed 0.25 g COD/g VSS. The response of whey digestion to several process parameters was investigated. Without pH-control, over 97% COD removal was obtained for influent concentrations from 5 to 28.8 g COD/1 and HRT of 5 days. However, instability was observed when the influent concentration was increased to 38.1 g COD/1. Gas production from whey is affected by organic loading rate (OLR). At an OLR less than 4 g COD/l-d, higher influent strength resulted in a higher methane production rate. When the OLR was greater than 6, higher strength feed or shorter hydraulic retention time (HRT) produced less methane. From the profiles of substrate concentration measured at various levels above the bottom of the reactor, two reaction stages, acidogenesis and methanogenesis were distinguished. It was experimentally illustrated that the rate of acidogenesis is much faster than the rate of methanogenesis in a whey anaerobic digestion system. The accumulation of VFAs in the first stage being faster than its assimilation in the second stage creates a distinct acidogenic phase in the bottom of the reactor. The instability caused by high influent concentration could be attributed to the accumulation of VFAs beyond the assimilative capacity of the methanogenic stage. A set of empirical models for accumulation and degradation of VFAs was developed using linear regression analysis. The requirement for maintaining this system in a dynamic balance was that the degradation capacity for VFA in the second stage be greater than the accumulation of VFA in the first stage. Based on this idea, the optimal influent concentration was given as between 25 to 30 g COD/1 for system stability. A hypothesis was proposed in this study that a proper amount of sulfate may be applied to moderate the detrimental influence of excess hydrogen on a stressed anaerobic reactor. The effect of sulfate was tested to study the biochemical mechanism. The permissible influent COD concentration was increased from 30 g COD/1 to 50 g COD/1 by using sulfate addition. The pH in the reactor was on the average 0.8 units higher and the concentration of butyric acid in the acidogenic phase much lower with added sulfate than without sulfate addition. The significant improvement of process stability and treatment efficiency made by the addition of sulfate clearly illustrated that sulfate acted like a stimulator which helped to maintain conditions favorable to methanogenesis. The mechanism of this stimulation is explained according to thermodynamics and hydrogen regulation which suggested that sulfate is able to promote the β-oxidation of VFAs by consuming hydrogen. A two-stage inhibition mechanism was proposed to explain the inhibition of high VFA concentrations and the stimulation of sulfate. Higher hydrogen pressure is the cause of preliminary inhibition, resulting in the accumulation of VFAs, which subsequently inhibit the activity and growth of methanogens in the second inhibition stage. The mechanism of inhibition of methanogens from VFAs was interpreted as being caused by the acidification of the internal cytoplasm and destruction of the pH gradient by non-ionized acids based on the theory of bacterial membrane transport. A new control strategy for stabilization of an anaerobic system is recommended. Under the optimal operating conditions based on the results in the first three steps, over 97% reduction of COD was achieved when the influent COD was 30 g /l using an HRT of 2 days, an OLR of 16.61 g COD/l-d and sulfate concentration of 0.2 g/1. / Applied Science, Faculty of / Chemical and Biological Engineering, Department of / Graduate