Trouble with Mycobacterium marinum!? - Microbiology: bacterial culture (Feb/15/2006 )
Can anyone give me some advice on the best way to grow Mycobacterium marinum in liquid culture? We have a wild type strain in our lab which can be successfully grown on Difco Middlebrook agar plates. However, when it it transferred into liquid culture (Difco Middlebrook 7H9 media supplemented with Middlebrook ADC enrichment) the bacteria do not grow. Does anyone have any suggestions. Bring back E. coli all is forgiven!
[quote name='Charlie Ellis' date='Feb 15 2006, 01:48 PM' post='41246']
I found 2 articles of ur interest; some times some physical parameters can be the responsibles for the non-growing bacteria like pH of the water...
"Individual isolates of M. marinum 1218R were streaked for single
colonies on 7H10 medium (Difco, Detroit, Mich.) supplemented with 10%
Middlebrook oleic acid, albumin, dextrose, and catalase enrichment (OADC;
Difco) and grown at 328C for 5 to 7 days. A single colony was inoculated into 7H9
medium (Difco) supplemented with 10% OADC and 0.2% Tween 80 and allowed
to incubate at 328C for 10 to 14 days. The cultures were then frozen in
0.5-ml aliquots at 2708C in 7H9 medium with 10% OADC."
"All M. marinum strains were grown in Middlebrook 7H9 medium supplemented with Middlebrook OADC (10% oleic acid-albumin-dextrose complex; Becton Dickinson, Cockeysville, Md.) and 0.5% glycerol. Plates were solidified with 2% agar. Cultures of M. marinum were prepared by inoculating 10 ml of medium in a tissue culture flask from the freezer stock, incubating at 30°C for 1 week, and storing at 4°C for several weeks. To produce rapidly growing M. marinum for all experiments, the stored culture was diluted 1/10 in fresh medium and incubated at 30°C for 3 days prior to infections."
Thanks for information! It's really appreciated. I'll will have a look at it and see what I can do!!