Hello, people that view this Instructable.
Here is an attempt to make a green bioinsecticide from Bacillus L from soil samples. To see how to take samples from soil Please see my Extracting bacteria from soil, etc for more detail. Cry A gene from this genius is very special it ONLY targets specific insects.
For example, Cry2ab and Cry3ab toxin are very toxic to ants while others have selectivity to beetles and other insects like Mosquitoes. I have used a good 1000 to 4000 Zero to 60 min centrifuge from eBay to do this. I have bought chemicals and supplies (centrifuge tubes) to do this as well to get a pure sample and kept them frozen to preserve them.
My first attempt was on Ants and the medium was a Lemon mix cake with 10 mM per L Potassium bismuth iodide (KBiI4) that reduced other bacteria from growing. It worked quite well I got cultures and spun them down. The solution test on ants on my property was a light brown solution.
Step 1: Hazards With Compost and Bacteria.
Although it very uniquely please be careful with an unknown bacteria or microorganism. Also, Mold on compost can grow which can be hazardous. If you see black or red mold pour bleach into it to kill those spores and DO not disturb the fungi!!!! This will neutralize the toxins and it safe now.
If you have a weakened immune system do not attempt to do this OK. Fair warning with bacteria.
As said never grow anything that pathogenic to yourself it also Illegal.
Step 2: Growing First Culture.
First in a large bottle with Lemon pie crust sugar and Potassium Bismuth Iodide I grew the cultures. Insulated them and put them into bottles. They were grown and filtered once and then sent with iron chloride which turned them black to a filter. 1 g of iron chloride turned black due to some tannins present.
Then they were centrifuged at 3000 RPM for 10 minutes.
The result was a light brown black material which had biofilm which I tested on Ants. I will tell you later if it worked well.
Step 3: Final Prep of Insecticide for Fruit Flies From Sugar, Cocoa and Milk Powder.
A mixture of 10 g cocoa, 10 g milk sugar, and 5 g sugar was added to 350 ml of water with culture and allowed to grow for 2-3 weeks until clear biofilm could be seen. By the pics, you can see the pellets that are black (impurities that have been purified by centrifugal force.
It was cleaned that way and put into the freezer at -10 degrees C to freeze the liquid and preserve the sample. 3000 RPM for 20 minutes was essential to remove most of the impurities. However the max volume I got before I added it to the bags was 350 ml plus or minus 1 ml. So I lost 14 percent of the sample that way but the loss is not that much but still a loss.
Biofilm from 3 weeks and an odd smell were obvious with the culture at 20 degrees C per day. Increasing the temp with Urea and Protein mix did not do anything. See the next Step.
Step 4: Growing Same Bacteria With Urea and Cocoa Hemp Protein Mix for 3 Weeks.
Good amount of 10 g Urea and 10 g Protein mix was added to 400 ml of tap water with bacteria to produce the culture. It was allowed to grow a few days.
Trouble arises when I used a green light halogen it got too hot too much and the bacteria slowed down and took 5 months to grow. It was too slow so it would be useful. It better just with normal temps. Maybe the ammonia produced from Urea and Nitrogen source harmed the bacteria producing waste and spores that stressed the bacteria out.
Some good lessons learned about this method too.
Step 5: Ant Affect and Fruit Flies Affect With Cultures.
These were some of the results.
The banana culture compost and regular compost did not stop the fruit flies with this Insecticide. I had issues with the ants too. Also, I think although it was well cultured it was the wrong type of Cry A gene. It was neither Cry2ab or Cry3ab and maybe fruitflies are immune to this toxin as some research said.
Although it failed I got a useful info and learned how to grow successfully bacteria cultures. Even if it did not work I know why and can explain it.
Once again thank for viewing this Instructable!