Session Title: Global Climate Change
Presentation Date: Thursday, August 21, 2008
Poster Board Number: 0358
DEVELOPMENT OF A METHOD FOR ISOLATING VIRUSES OF RUMINAL METHANOGENS
D. Ouwerkerk, E. Constantinoiu, A. Klieve
Department of Primary Industries and Fisheries, Animal Science, Moorooka, Australia
Background and aims: Enteric methane generation in ruminants is a significant source of greenhouse gas emissions. While bacteriophages have been used to lower populations of bacteria such as Escherichia coli O157:H7 in ruminants, no viruses of rumen methanogens have been isolated. The objective of this study was to develop methodologies to isolate lytic viruses of ruminal methanogenic archaea.
Methods: A modification of the Hungate roll-tube technique has been developed. The roll-tube technique allows for pressurisation of the tube with H2 gas which is necessary for the growth of the methanogens. Methanobrevibacter sp. YE286 was inoculated into 50 mL BY+ broth medium and harvested in early log growth phase by centrifugation to concentrate the cells. These cells were inoculated into the modified roll tube, pressurised to 30 psi with H2 and incubated at 39 oC. Standard techniques for the concentration and isolation of bacteriophage populations from a variety of sources including abattoir wash, sewage and rumen fluid will be used for isolation of methanogen viruses.
Results: A thick lawn of methanogenic bacteria has been successfully grown in the modified roll tubes and source material obtained for concentration of viruses for screening against Methanobrevibacter sp. YE286.
Conclusion: The establishment of a visible lawn of Methanobrevibacter sp YE286 using a modified roll-tube technique will greatly enhance efforts to isolate viruses that may reduce methanogen populations and methane emissions.