[No abstract submitted] / Science, Faculty of / Zoology, Department of / Graduate

Thesis (M.Sc.(Agric.))(Animal Science)--University of Pretoria, 2007. / Summaries in Afrikaans and English. includes bibliographical references. Available on the Internet via the World Wide Web.

Thesis (Ph. D.)--Columbia University, 1925. / Vita. Bibliography: p. 28-29.

Thesis--Copenhagen. / "Litteratur": p. [243]-249.

Thesis--Copenhagen. / "Litteratur": p. [243]-249.

Thesis-Uppsala. / Summary in English. Bibliography: p. 184-188.

To avoid environmental pressure from water authorities, specifically regarding nitrogen and sulfate limits in tannery wastewater, modifications to existing deliming processes have been made. Conventional ammonium salt deliming methods contribute to Total Kjeldahl Nitrogen values in the region of 0.5 – 1.0g/L (33-67% of total TKN). Sulfate levels are increased with the use of organic deliming and ammonium sulfate deliming to the extent of 0.9g/L (27% of total sulfate). To understand the dynamics and kinetics of carbon dioxide equilibrium, the movement of carbon dioxide into deliming water, through carbonic acid, bicarbonate and ultimately into carbonates at liming or early deliming pH was studied. It was shown in this study that effective lime removal, at optimum conditions, resulted in fully delimed pelts at highly comparable quality and times compared to conventional ammonium salt deliming

A polarimetric method for monitoring the rate of soluble collagen breakdown by collagenase enzyme action has been developed. The method represents an extension of previous physicochemical techniques based on viscometry, but is simpler and easier to carry out, particularly in the case of reaction rate studies. The method was developed arising from reports of collagenase activity measurement on inappropriate substrates such as gelatin, modified collagens and synthetic polypeptides. The optical method depends on measurement of the loss in optical rotation in solutions of soluble calfskin collagen resulting from initial enzymic cleavage of the collagen trip1e-helix, followed by spontaneous unwinding of the resultant unstable helical fragments. Specific assay conditions were chosen to ensure that the loss in optical rotation following enzymic cleavage was rapid and complete. The method is specific since in the absence of collagenase, non-specific proteinases produce only a limited decrease in solution optical activity. The method has also been compared with established physicochemical assay techniques and compares favourably with both viscometric and titrimetric collagenase assays. The availability of a rapid, sensitive and quantitative procedure for measurement of collagenase activity provides a convenient means for detecting the presence of collagenase in solution and examination of hide bacterial cultures for collagenase production. In addition, a study of biocidal compounds of potential interest in hide preservation for possible inhibitory effects on collagenase is conveniently carried out with the method. Fundamental research into synergistic action in enzymic hydrolysis of collagen is now possible, providing valuable insight into the mechanism of raw hide biodeterioration.

In the leather industry fungicides are often used for the protection of wet-blue leather. These fungicides are usually only sparingly soluble and are therefore formulated together with surfactants in order to increase their solubility and to ensure an even distribution over the surface of the hide after treatment. Solutions containing both fungicides and surfactant are complex. The nature of these solutions was investigated. By means of UV/Vis spectroscopy and viscometry it was shown that the surfactant and fungicides form micelles and mixed micelles in solution. The nature of these micelles and mixed micelles was dependent on the solution temperature as well as on the concentrations of the surfactant and fungicides. At the higher temperatures and concentrations transition to large, possibly rod-shaped, mixed micelles occurred. The interaction between the selected fungicides 2-(thiocyanomethylthio)benzothiazole and n-octyl-4-isothiazol-3-one with bovine skin collagen in the form of both limed and lightly chromed hide powder in the presence of the non ionic surfactant Triton X -100 was investigated. Fungicide uptake was determined by difference measurements on the float solutions at regular intervals during treatment. Binding was rapid with equilibrium being established within the first six hours even for the solutions with the highest surfactant concentration. Binding failed to follow a normal mass-action binding-type isotherm approaching a saturation limit, but increased continuously indicating a co-operative effect whereby binding site affinity actually increased with the amount of ligand bound. Binding was accompanied by a drop in the free surfactant in the solution at the higher biocide levels indicating the formation of complex mixed micelles which bind to the collagen fibres. The uptake and antifungal activity of commercial fomulations of the fungicides on chrome-tanned wet-blue leather was investigated at various treatment temperatures. At lower fungicide treatment concentrations, binding tended to follow a typical mass-action type binding isotherm, increasing slightly with temperature. At higher float concentrations, an inflexion point was apparent beyond which uptake showed a marked increase with concentration. This inflexion point, signifying a change in binding characteristics, occurred at progressively lower concentrations with increasing temperature. Antifungal activity in terms of storage periods to onset of fungal growth was determined on the wet-blue leather cuttings immediately after treatment and drainage and also on sample discs after exhaustive extraction of free fungicide using dichloromethane. Storage performance testing of the various treated wet-blue leathers was carried out by different methods. Residual protective periods showed a curvilinear increase with dosage offer and surface uptake. In the low dosage range treatment temperature had only a relatively slight effect in promoting uptake and improving storage protection. At higher dosages, the influence of temperature on uptake and storage protection was greater due to the increase in surface binding of the fungicides at the elevated temperatures. Only a portion of the fungicide uptake was recovered by direct solvent extraction of the treated wet-blue leather. Solvent extraction reduced storage margins. The storage response in relation to fungicide content was, however comparable after extraction, indicating that both irreversibly bound and physically associated fungicide offered effective protection. Results of the study provide further insight into the mode of interaction of fungicide emulsion dispersion with bovine skin collagen, and the importance of the emulsion dispersions and its stability in determining the uptake of fungicide.

Animal hides are the basic raw material of the leather industry and they undergo rapid putrefaction unless "cured". This study investigated the role and interactive effects of three selected bacteria, Pseudomonas aeruginosa. Clostridium histoly ticum and Clostridium sporogenes in in-situ cattle hide degradation using a model system set up for the purpose. The system consisted of 3cm diameter hide pieces contained in sealed jars and sterilised by ethylene oxide to remove resident microbes and inactivate autolytic tissue enzymes. The inocula were prepared either as individual cultures or as combinations of two inocula or all three inocula. Degradative changes during storage at 30°C were measured for up to 8 days using ten different parameters. Initial trials confirmed that the selected inocula were readily isolated from raw hides and could outcompete resident populations to produce putrefactive decomposition. Growth rates and enzyme profiles of the organisms and the effects of nutrients and reductants on their relative denaturative effects were used to standardise the system. Trials on the effects of ethylene oxide indicated the suitability of the method for hide and collagen sterilisation. The findings of in-situ trials with the selected inocula confirmed previous studies of protein putrefaction in that a bacterial succession was evident involving aerobic proteolytic bacteria, micro-aerophilic proteolytic bacteria and strictly anaerobic amino acid degrading bacteria. However, this study showed that the micro-aerophilic collagenase producing C. histolyticum degraded hides at a far greater rate when inoculated on its own than when in the presence of either or both of the other two inocula. It also demonstrated a bacterial antagonism between the two clostridia in which C. sporogenes prevented degradative changes occurring for up to 4-6 days possibly due to cysteine production by C. sporogenes. These findings have implications for hide preservation since maintenance of aerobic conditions and suppression of spore outgrowth could be used to delay growth of collagenase producing clostridia. The use of C. sporogenes as a biocontrol agent is also postulated. The model system was also used to examine salted hides during storage and these studies indicated that Halobacteriaceae do not produce collagenase but that inadequately salted hides could possibly be subject to degradation by delsulfovibrios.