Help with sporulation of Bacillus globisporus

[Enigl]

I have used this philosophy to get _Bacillus_ to sporulate:

1.  Optimum growth temperature and "plain old" nutrient agar (little or no
stress on cells).  Be sure the agar has a very solid surface. 

2. Then, use  surface inoculation:  Spread plate technique with a cool
glass rod. Heavy inoculation will crowd the developing colonies and they
overlap.  (Gradual increasing stress on cells).

3.  Let the medium dry out in the incubator and lower water activity from
0.99 to 0.92 this should also be gradual stress.  Give them time to
trigger spores.

Other tricks are to increase stress by sub or super optimal temperatures
during the dry-down process.



Davin


Davin C. Enigl, MS-MEAS, President-Microbiologist
HACCP Validations-sm  Hazard Analysis and Critical Control Points for the
Food, Cosmetic, Pharmaceutical, and Nutritional Supplement Industry
Voice: (916) 989-8264,  Fax: (916) 989-8205,  Pager: (714) 725-7695
9040 Erle Blunden Way
Fair Oaks, CA 95628
September 18, 1996
10:48 am

In article <3238BE20.3173 at pop.fast.net>, Brian Dale <briand at pop.fast.net>
writes:
>Subject:	Help with sporulation of Bacillus globisporus
>From:	Brian Dale <briand at pop.fast.net>
>Date:	Thu, 12 Sep 1996 21:51:28 -0400
>
>Hello,
>
>I work for a small biotech company and we've recently begun working with 
>the low-temperature bug B.globisporus.  The only trouble is, we can't 
>get it to spore.  We've done research and found that there isn't much 
>out there we could find on this organism.  Anybody have any suggestions? 
>We've been using a basic sporulation media (ie, Shaffer's Sporulation) 
>with few modifications with no luck.  
>
>Hope you can help!
>
>Brian
>
>