DIY DIY Culture Bacillus amyloliquefaciens (active ingredient in products like Hydroguard)

[Bob's Auto's]

Hello fellow growers and DIY's.

Last winter I found a thread on Reddit about culturing your own Bacillus amyloliquefaciens and thought "how hard can that be?"
Well not that hard at all!! Actually it was a piece of cake

Let me start by listing all the supply's you need...

• One bottle of Hydroguard. 125ml will suffice and will probably set you up for life.
• Or if you are unable to get your hands on Hydroguard like me you'll have to go search the net for a 'spore' version of the Bacillus. I found mine on Ebay.
• Two cans of chickpeas (the cheapest you can find, because those will have lots of broth inside).
• A bottle
• A jar or other glass container (approx. 500ml)
• Optional, but recommended, a magnetic stirrer with hotplate. I bought mine on Ali.
• One magnetic stir bar

Let's start cookin'...

• Start by filling the bottle with hot water (>76°C or >169°F). I have a Cooker fossette in the kitchen so mine comes out boiling. After put the cap on and set aside.
• Sterilize the glass jar/measuring cup. --> put some water in it and put in the microwave for 5min. Be very careful when you take it back out, because it can scald you!! Remove the water that's left inside and wrap a sheet of aluminum foil over the top. This is to prevent fungal spores to enter the container. Don't use a screw lid as this will create a vacuum...
• Open the chickpeas cans and quickly transfer the broth into the sterilized jar and cap with the aluminum foil again. I used the chickpeas for a delicious mutton curry, yummy
• In case you bought the bottle of Hydroguard open it up and put a couple of drops into the broth after which you cap the container with the foil. Since I only had access to the spores I used the scoop that came with it and put five scoops into the broth. I weighed one scoop and this amounted to 0.01gr. So I used 0.05gr of the 10 grams I bought. This is also going to last me a long long time...
• Sterilize your magnetic stir bar with a alcohol wipe and put into the broth. Again capping the vessel with the foil
• Put the glass container on the heated stir plate and set it to 36°C/97°F and turn the stirrer on (approx. 240 rpm)
• Now comes the hard part. Wait....And wait some more. Wait at least 6 hours, I did 18 as I wanted to see what would happen. Edit: I just made more bacteria
• After 18 hours turn of the magnetic stirrer, remove some of the water from your bottle (approx. 300ml, because that's about the amount of broth you recovered from the chickpeas) and transfer the culture you've made into the bottle with water diluting it three times. (be careful not to lose the magnetic stir bar)
• Cap the bottle and let it rest for a couple of hours.
• How to know it worked... Take a whiff from the bottle. It should smell like fresh sweat. Ok, not the most likeable smell, but one you should recognize. If the smell is foul or rotten you've had a contamination and you'll have to start from the beginning.

Congratulations! You've now made a bottle of Bacillus amyloliquefaciens with a higher cfu count then the original, actually about 1 x 10⁸ cfu's/ml, that's 10.000x stronger!

Bacillus amyloliquefaciens (strain IT35)

I used five scoops, but if using a longer incubation time one should have done the trick

The weight of one scoop

The broth mixed with the bacillus spores on the magnetic stirrer set to 36°C/97°F and approx. 240 rpm

Just mixed the culture with the remaining water from the bottle and set to rest

After four hours of resting I saw the bacteria settling on the bottom. I used a flashlight so it was easier to photograph

Regards,

Bob

p.s. I would like to thank user 53003_420 on Reddit for supplying this information.

 

[Bob's Auto's]

I made a couple of microscope shots of the culture today.

Magnification 64x - large colonies Bacillus amyloliquefaciens

magnification 160x - small colonies Bacillus amyloliquefaciens

Magnification 640x - single Bacillus amyloliquefaciens

 

[Death The Cultivator]

I made a couple of microscope shots of the culture today.

Magnification 64x - large colonies Bacillus amyloliquefaciens
View attachment 1299443

magnification 160x - small colonies Bacillus amyloliquefaciens
View attachment 1299446

Magnification 640x - single Bacillus amyloliquefaciens
View attachment 1299448

Can I do this with other innoculants?

 

[Bob's Auto's]

Can I do this with other innoculants?

From what I've been reading it is possible to do this with any Bacillus type, but if you have multiple strains in your base. I think the strongest will prevail and consume the weaker strains

 

[Death The Cultivator]

From what I've been reading it is possible to do this with any Bacillus type, but if you have multiple strains in your base. I think the strongest will prevail and consume the weaker strains

Well nevermind then lol.

 

[Bob's Auto's]

Well nevermind then lol.

Penny for your thought? What were you thinking about? I might be of some help

 

[Death The Cultivator]

Penny for your thought? What were you thinking about? I might be of some help

I have a an N and P fixing innoculant with several strains each.

 

[Bob's Auto's]

Which brand? Or which strains?

 

[Death The Cultivator]

Which brand? Or which strains?

 

[Bob's Auto's]

I wouldn't recommend making a culture with either of them.

Both pictures are a mixture of fungi and bacteria with added 'flavoring'. The fungi will die and be eaten by the bacteria and then the bacteria will fight for survival with the outcome that (probably) Bacillus megat(h)erium will be the sole survivor. Now megatherium is the bacteria you want for protection against all outside bad things like mold and even the larvae from the moss fly or thrips. It produces it's own form of penicillin. Cool things those bacteria

The reason they can be in the same bag or pot is that they are in the so called 'spore' version. Just add water and they will hatch so to speak... Once in the ground they will compete with each other until the food for the bacteria is gone and they die off and then the fungi will take over.

Regards,

B