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ARS Home » Southeast Area » Stoneville, Mississippi » Warmwater Aquaculture Research Unit » Research » Publications at this Location » Publication #230628

Title: Improved Pulsed-Field Gel Electrophoresis Procedure for the Analysis of F. columnare Isolates Previously Affected by DNA Degradation

Author
item SOTO, E - MISS. STATE UNIVERSITY
item LAWRENCE, M - MISS. STATE UNIVERSITY
item MAUEL, M - MISS. STATE UNIVERSITY

Submitted to: Veterinary Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/1/2008
Publication Date: 3/1/2008
Citation: Soto, E., Lawrence, M., Mauel, M.J. 2008. Improved Pulsed-Field Gel Electrophoresis Procedure for the Analysis of F. columnare Isolates Previously Affected by DNA Degradation. Veterinary Microbiology. 128:207-212.

Interpretive Summary: Pulsed-field gel electrophoresis (PFGE) has previously been shown to be capable of distinguishing genetic grouping of Flavobacterium columnare. However, some isolates of F. columnare where not able to be analyzed by the earlier protocol. In the current study, DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. Utilizing the new protocol PFGE analysis is shown to be a reproducible highly discriminating epidemiological method for studying F. columnare isolates regardless of fish host.

Technical Abstract: Flavobacterium columnare is a fresh water bacterium that causes columnaris diseases in over 36 fish species. Intra-species typing of F. columnare can be performed by pulsed-field gel electrophoresis (PFGE). However, this method is hampered by the degradation of chromosomal DNA in about 10% of strains. In the current study, DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. The results substantiate that after problems due to DNases are overcome, PFGE analysis is a reproducible highly discriminating epidemiological method for studying F. columnare isolates regardless of fish host.