Check out The Prokaryotes, it has a lengthy section in Bacillus spp. and
sporulation/germination. In most cases, the addition of Manganese will
improve sporulation. Spores a generally found in older ciltures, so let them
grow a while and then you can try to isolate spores.
A general schema would be to treat the samples you want to enumerate with
Ethanol (filter sterilized) for up to 1 hour. Then plate out aliquots from
that . EtOH will kill almost all vegetative cells after 1 hour. Then as the
plates grow up you can get CFU's from what grows after such treatment.
CSU San Marcos
Walter Blank wrote:
> Hi all,
> I'm trying to find procedures for inducing sporulation in Bacillus (B.t.
> or subtilis), separating out vegetative cells, and enumerating the
> spores by cfu's (so I guess they'd have to be effectively germinated).
> If anyone could pass along or point me in the direction of favorite
> methods I'd appreciate it.
> Walter Blank