Improved pulsed-field gel electrophoresis procedure for the analysis of Flavobacterium columnare isolates previously affected by DNA degradation.
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.
Soto
Esteban
E
Thad Cochran National Warmwater Aquaculture Center, Stoneville, MS 38776, USA.
Mauel
Michael
M
Lawrence
Mark
M
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods / Q000662:veterinary*; D005393:Fish Diseases / Q000382:microbiology*; D005399:Fishes / Q000382:microbiology; D045826:Flavobacteriaceae Infections / Q000382:microbiology / Q000662:veterinary*; D005417:Flavobacterium / Q000145:classification / Q000235:genetics*; D005557:Formaldehyde / Q000737:chemistry; D014644:Genetic Variation; D010802:Phylogeny
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005557:Formaldehyde; C035041:endodeoxyribonuclease MluI; D015252:Deoxyribonucleases, Type II Site-Specific