Novel Systems for Bacterial Preservation and Enumeration

• Main Author: Hays, Henry Charles Wilson
• Published: University of Leeds 2003
• Subjects: 579
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.491750

Much like books in a library, microbiological strains must be able to be stored over prolonged periods of time, while maintaining their condition and accessibility for the purposes ofresearch and reference. Many methods currently employed for the storing of microorganisms require highly specialized storage systems; substantial technical equipment; know-how and temperature controlled environments. Whilst these methods are effective, it is accepted that they can be expensive and technically demanding. Traditionally, methods ofevaluating novel storage techniques require the use of classical bacterial enumeration techniques, which are slow, labour intensive and expensive to automate. This work investigates the use of in-vivo bacterial bioluminescence as an alternative bacterial enumeration tool to aid the identification of novel preservatives and preservation formats for the quantitative and qualitative preservation of bacteria. The development of a cheap, simple, yet effective storage alternative to current preservation systems based on a novel self-drying concept, containing activated charcoal and sucrose protectant is reported. In addition, the evaluation of DNA transformed bacteria using plasmids containing lux gene cassettes was studied in order to assess transformation efficiencies in competent cells. Currently, competent cells are distributed and stored at very low temperatures in order to maintain their viability and competence, therefore, the demand for a technology that will alleviate the need for handling and storing of competent cells at sub-zero temperatures· is certainly present. However, methods employed in screening and developing new competent cell formats often rely on classical enumeration techniques. Therefore, the development of a fast screening tool in the form of bioluminescence would aid the development of novel competent cell formats. To this aim, a fast screening tool for the identification of positive competent cell. formulations was investigated.