Improving the culturability of freshwater bacteria using FW70, a low-nutrient solid medium amended with sodium pyruvate.
Bacterial culture based on the use of plate media is an effective method for investigating bacterial populations in the environment. To improve the culturability of bacteria from freshwater lakes, we developed a new medium, FW70, which contains sodium pyruvate, casamino acids, and lake water and is solidified using gellan gum. To test the importance of these components, we prepared a series of media in which one or more of the components was absent. Water was sampled 31 times from 3 Japanese lakes and was passed through a membrane filter (pore size = 0.45 microm) to remove fast-growing microbes before the water was spread onto the plates. In most cases, significantly larger numbers of bacterial colonies were detected on FW70 than on other media. Furthermore, to test the practicality of FW70, we compared it with standard nutrient agar and R2A agar. In all cases, the culturability was significantly greater on FW70 than on standard nutrient agar or R2A agar. Some isolates recovered by means of FW70 belonged to bacteria that had not previously been classified. These results suggest that FW70 improves the culturability of freshwater bacteria and can be used for the isolation of novel bacteria as a result of the filtration step.
Imazaki
Iori
I
National Agricultural Research Center, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan. iiori@affrc.go.jp
Kobori
Youichi
Y
D000596:Amino Acids / Q000378:metabolism; D001419:Bacteria / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D003470:Culture Media / Q000737:chemistry*; D005374:Filtration; D005618:Fresh Water / Q000382:microbiology*; D007564:Japan; D011135:Polysaccharides, Bacterial; D011773:Pyruvates / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012964:Sodium / Q000378:metabolism; D019346:Sodium Acetate / Q000378:metabolism
D000596:Amino Acids; D003470:Culture Media; D011135:Polysaccharides, Bacterial; D011773:Pyruvates; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C017721:casamino acids; D019346:Sodium Acetate; C048288:gellan gum; D012964:Sodium