Microbial Pesticides

 Microbial Pesticides are pest control agents that can be derived from virus, fungi, bacteria, or
protozoa. The most commonly used commercial species pesticide is Bascillus thuringiensis (Bt).
 Advantages:
1. Highly specific (only toxic to certain types of insect)
2. Little effect on non-target and beneficial insect
3. No effect on human
4. Insect have not so far shown resistance to pest control agents.
 Disadvantages:
1. Killing time is slow compared to chemical pesticides
2. Some are unstable when released into the nature
3. Production is expensive
 Production of Bacillus thuringiensis bio pesticide using commercial lab medium
A. Main material: Bacterial strain Bacillus thuringiensis sv tolworthi, strain 344, belonging to
Embrapa Maize and Sorghum Microbial Bank which isolated from a soil sample.
B. Procedure
1. Media preparation: Strain 344 was sub cultured in three different media:
a. Medium 1 was composed of lab commercial Luria Bertani (LB) plus salts and
glucose.
b. Medium 2 was composed of glucose, soybean flour plus salts.
c. Medium 3 was composed of liquid swine manure (LSM).
2. Seed culture preparation
3. Growth of B. thuringiensis tolworthi in three different media
The appropriately diluted samples were plated on agar plates and incubated overnight at
30oC to form fully developed colonies.
4. Cell mass
Samples of 100 mL were centrifuged at 10000 rpm for 20 minutes. Dry weight was
calculated and expressed in gram per liter.
5. Spore count
Plates were incubated at 25 ± 1oC for 24 hours to form fully developed colonies that were
counted and expressed in colony forming units per mL (c.f.u./mL).
6. pH adjustment
The pH of all media was adjusted to 7.2 with NaOH after sterilization.
7. Toxicity test
The spore crystal complex produced in different media was assayed against 2 day old
larvae Larval mortality was scored up to 4 days after larvae infection to check
pathogenicity.
MSG CSPT
 How does Bt work?
1. Insect eat Bt crystals and spores
2. The toxin binds to specific receptors in the gut and the inset will stop eating
3. The crystal cause the gut wall to break down, allowing spores and bacteria to enter the body
4. Insect dies as spore and gut bacteria proliferate in the body.