Bacterial interaction in hide biodeterioration with special reference to selected Clostridium species
- Authors: Thompson, Gillian Ann
- Date: 1995
- Subjects: Hides and skins -- Preservation Aerobic bacteria Pseudomonas aeruginosa Clostridium Halobacterium
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4041 , http://hdl.handle.net/10962/d1004102
- Description: 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.
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- Date Issued: 1995
A possible mechanism for enzymic depilation of skins
- Authors: Brady, Dean
- Date: 1989
- Subjects: Chemistry, Technical , Tanning
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3877 , http://hdl.handle.net/10962/d1001611
- Description: Streptomyces fradiae is a bacterium which has been previously found to produce extracellular enzymes which are capable of wool degradation and skin depilation. Streptomyces fradiae 3739 and other strains of Streptomyces were found in this study to be able to degrade a keratin source (wool) to a considerable degree. However according to the evidence of SEM micrographs presented here the highly keratinised spindle cells of the paracortex are fairly resistant to protease attack, and it is the cementation material which binds these cells together which is initially degraded by the proteases. A large degree of correlation was found with the strains of Streptomyces studied, between the ability of the individual strains to degrade wool and the ability of their extracellular proteases to reduce the depilation load of sheepskins. With further analysis S. fradiae 3739 was found to produce at least one amylase and four or more proteases. The proteases as a group had maximal proteolytic activity in the 8.0-9.0 pH unit range, and were considerably thermostabilised by the inclusion of calcium ions into the reaction solution. The protease group was found to cause depilation of merino sheepskins. For comparative purposes a protease produced by a strain of Proteus vulgaris isolated from a staling hide with hair slip (natural depilation) was studied. The protease activity was maximal in the alkaline region between 8.0-9.0 pH units. Tbe protease appeared to be a single enzyme with a molecular mass of approximately 44 000 daltons. The protease was maximally active at 40°C, although it was only thermostable to 30°C. The enzyme was ineffectual as a depilant except when the skin was pre-treated with a strong alkali, preferably including sodium sulphite in the protease preparation. One of the most important differences between the extracellular proteases of S. fradiae and P. vulgaris was that the former were greater in variety and caused a greater decrease in the depilation load of sheepskins than the latter. Further research with mixtures of commercial proteases provided evidence that a synergistic depilatory effect occurs when proteases of complementary bond specificities are used in conjunction in enzymic depilatory preparations. Some form of strong alkali treatment of skins was found to be necessary to produce leather of the prerequisite quality when the skin was depilated by proteases, otherwise the skin was found to be depleted and stiff. Calcium hydroxide alone was found to be inadequate for this task, probably owing to the fact that it is less alkaline than the lime-sulphide mixture. The calcium hydroxide (lime) must therefore be used in conjunction with sodium hydroxide (which makes the solution as alkaline as that of the lime-sulphide solution) to produce leather comparable to that produced by the lime sulphide treatment. A combination of the information provided by the present research and that gleaned from the relevent literature allows for the construction of a model to represent the possible mechanism of enzymic depilation of skins, in which depilation is caused by the disruption of the basement membrane at the dermal-epidermal junction by the degradation of its constituent molecular components by general proteases, resulting in the removal of the epidermis and its associated wool or hair
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- Date Issued: 1989