# How do you culture Certan (Bacillus thuringiensis)



## Michael Bush (Aug 2, 2002)

Well, I finally found it.
http://www.motherearthnews.com/menarch/archive/issues/074/074-111-01.htm 

They weren't doing it for wax moths, but I suppose the same system will apply. The problem is that once I have sprayed the frames, I never see any wax moths, and this method expects you to get some infected larvae.


----------



## magnet-man (Jul 10, 2004)

Michael, I am not sure that you will be able to culture Bacillus thuringiensis from B401. I believe B401 does not have any active spores and only contains the delta-endotoxin that B.t.a. produces. I could be wrong. When I get a chance I will check it under my microscope.

The B.t. that works well on wax moths is getting hard to find in the US, because it is more effective to insert the gene into crops. There is one company that produces a commercial wettable product that does contains the strain of B.t. that you are looking for. If you want, I can track them down for you.


----------



## Michael Bush (Aug 2, 2002)

>I believe B401 does not have any active spores and only contains the delta-endotoxin that B.t.a. produces.
http://www.beekeeping.com/swarm/b401_us.htm 

"B401 is a selective biological larvicide, which contains delta-endotoxin and spores of Bacillus thuringiensis Berliner, Serotype 7"

>There is one company that produces a commercial wettable product that does contains the strain of B.t. that you are looking for. If you want, I can track them down for you.

That would be quite nice. The B401 version is quite expensive.


----------



## magnet-man (Jul 10, 2004)

Dam that is one small bug! Even under 1,000 magnification it is tiny. The spore and the protein cyst is about the same size and shape. Here is a link that shows the various agents of BT. http://www.natur.dk/images/m_m2_probbak_bacillus_thuringiensis.jpg 

The sample of B401 that I checked has a large percentage of delta-endotoxin cyst to spores, but the two look alike. So you should be able to culture it directly from B401. 

Well, I guess it is time to get the Petri dish out.


----------



## dickm (May 19, 2002)

I bought some Certan from Canada. Can't you just culture from that?

dickm


----------



## Michael Bush (Aug 2, 2002)

The method in the Mother Earth News was to spray the original formula on the larvae and use the mashed up larvae in the milk to culture it. I'm not sure the spores will activate in just the milk. They may require their host to do that.


----------



## Michael Bush (Aug 2, 2002)

Of course the mushed up host larvae may also add some nutrients to the milk.


----------



## magnet-man (Jul 10, 2004)

Dick, Certan and B401 are the same thing. I just prepared a culture using milk and straight B401. We should know in 48 hrs if you can use straight B401 and milk. 

If you are using already treated mashed up larva, do you really need the milk?


----------



## Michael Bush (Aug 2, 2002)

Just quoting from the above link. I don't know if you need it or not.


----------



## magnet-man (Jul 10, 2004)

So MB are you planning on going into small scale production?


----------



## Michael Bush (Aug 2, 2002)

Not really. I can't sell it, I'm sure. But I'd like to know how to make my own.


----------



## magnet-man (Jul 10, 2004)

MB that is what I thought. It would be nice to eliminate a small cost here and there. It is like using OA. OA is so much less expensive then Apistan strips, I am not sure that I will buy strips again.


----------



## GaSteve (Apr 28, 2004)

You may want to read these.
http://filebox.vt.edu/cals/cses/chagedor/btprimer.html 
http://maarec.cas.psu.edu/PDFs/Wax_Moth_pm.pdf 

There are many strains of Bt that are used for different things. According to the Penn State report, the Bt used most against caterpillars in general is "not fully effective" against wax moth. Maybe "not fully effective" is good enough -- I don't know. But the Bt variety in Certan is not the same as the most widely used varieties used against caterpillars in general as it produces slightly different toxins. The Certan variety is also found in the products Agree and Xentari. It did say that the Bt toxin was activated in the alkaline conditions in the insect's gut, which probably explains why it can be successfully cultured in milk.

I like the idea I read on Dave Cushman's site about putting tobbaco leaves on top of stored brood boxes. Or better yet firing blanks from a gun seem to make some adults moths die from shock. It may not be practical, but it would be fun.








http://www.dave-cushman.net/bee/waxmothtreatment.html


----------



## Michael Bush (Aug 2, 2002)

>There are many strains of Bt that are used for different things. According to the Penn State report, the Bt used most against caterpillars in general is "not fully effective" against wax moth. 
http://www.beekeeping.com/swarm/b401_us.htm 
"B401 is a selective biological larvicide, which contains delta-endotoxin and spores of Bacillus thuringiensis Berliner, Serotype 7, developed by Sandoz. B401® is selectively toxic to the greater wax moth, Galleria mellonella. Its efficacy is linked with the delta-endotoxin, which causes destruction of the mid-gut epithelium, after ingestion by the larvae. B401® gives effective, long-lasting control of susceptible larval stages in stored combs or active hives."


----------



## magnet-man (Jul 10, 2004)

Its ALIVE!!! I Dr. Frankensamples have created life from B401. Wait what is that noise? Oh my gosh, Its a mob with pitchforks and torches.

Yes Virginia you can culture Bacillus thuringiensis from the spores in B401.

I just checked my crude culture of BT that I made with a little bit of milk and it is loaded with bacteria. Unfortunately I did not sterilize my culture before starting, but the bacteria looks a lot like BT. I guess the next step is to do a proper culture.


----------



## Flyer Jim (Apr 22, 2004)

Question is: Is it still alive? I got my bottle of certan and a gallon of cow juice, now what? what's the recipe and how much do you dilute the culture? Did anybody try this and get it to work? Thanks for any information
Jim


----------



## Jim Fischer (Jan 5, 2001)

> The method in the Mother Earth News was to spray 
> the original formula on the larvae and use the 
> mashed up larvae in the milk to culture it. I'm 
> not sure the spores will activate in just the 
> milk. They may require their host to do that.

The point of using the infected larvae was to 
assure getting a significant percentage of the
exact strain that attacks the larvae in one's
culture. 

Just as it is difficult, perhaps impossible to
get a pure single-strain yeast to be the only
yeast in any loaf of bread made anywhere outside
a NASA clean room, your "culture" will have a
range of slightly different BT variants in it,
as well as a small amount of whatever spores
landed on your goo when you mixed it.

By culturing from infected (dead) larvae, you
are introducing a large number (millions,
maybe more) of spores from a strain that
grows on the larvae, which is what you want.

While you will never have a 100% pure culture
of a single strain, you will get an overwhelming
majority of the strain you want.

If milk works, great. If not, you can mail-order
pre-filled petri dishes of various types of growth
media, all sterile, and ready to use.


----------



## butterfly (Jan 6, 2005)

Mike Knows


----------



## Flyer Jim (Apr 22, 2004)

Thanks Jim & Butterfly. The last we heard from Mike and Magnet-Man on this thread they were standing around a cauldron waving an old issue of Mother Earth News shouting "double double toil and trouble". I was just wondering if they had come up with any new info on making your own BT. (not bee tea) Thanks again, Jim


----------



## bjerm2 (Jun 9, 2004)

Michael Bush 
I get some of my 'stuff' from Gardens Alive. They sell various bacteria that is mixed in with water and then sprayed on crops or trees. 
Another thing I use, like for Japanese beetles, is collect a few dozen and put them into a container with water. Let stand for a few days to ferment then strain it and use the spray on the trees. Works well. The fermenting makes up the bacteria that kills the adults.
Just my $0.02. Hope this helps.
Dan 

PS Take a look at this WEBB site
http://www.gardensalive.com/Default.asp?bhcd2=1109872720


----------



## Michael Bush (Aug 2, 2002)

I haven't had time to pursue it, but was planning on trying it come spring before the wax moths start reproducing.


----------



## magnet-man (Jul 10, 2004)

I am like MB. Just haven't had time to pursue it either.


----------

