How to cultivate Entomozan C against bedbugs? - briefly
Grow Entomozan C in a sterile glucose‑yeast extract broth at 25‑28 °C, pH 6.5‑7.0, with agitation of 150 rpm for 5‑7 days. Harvest the spores, dilute to 10⁶ spores ml⁻¹, and apply the suspension as a spray to infested areas.
How to cultivate Entomozan C against bedbugs? - in detail
Cultivating Entomozan C for effective bed‑bug control requires a sterile environment, precise nutrient composition, and strict temperature regulation.
Begin with a pure starter culture. Transfer a single colony from a cryopreserved vial onto a petri dish containing malt extract agar (MEA) supplemented with 2 % glucose. Incubate at 25 °C for 48 hours until the mycelium covers the surface.
Prepare a liquid fermentation medium. Dissolve 30 g of glucose, 5 g of yeast extract, 3 g of peptone, and 2 g of magnesium sulfate per liter of distilled water. Adjust pH to 6.5 with sterile 0.1 M NaOH. Autoclave at 121 °C for 15 minutes.
Inoculate the liquid medium with 5 % (v/v) of the agar-grown culture. Place the flask in a shaking incubator set to 150 rpm and maintain temperature at 27 °C. Monitor growth daily; optimal spore concentration is reached after 72 hours, indicated by a cloudy suspension and microscopic observation of abundant conidia.
Harvest spores by filtering the suspension through sterile cheesecloth. Rinse the collected material with sterile water containing 0.01 % Tween 80 to prevent clumping. Centrifuge at 3,000 g for 10 minutes, discard supernatant, and resuspend the pellet in a carrier oil (e.g., mineral oil) at a concentration of 1 × 10⁹ spores per milliliter.
Formulate the product for field application. Mix the spore‑oil suspension with a granular carrier (e.g., diatomaceous earth) at a ratio of 1 : 100 (v/w). Store the blend in airtight containers at 4 °C; viability remains above 90 % for six months.
Application guidelines: disperse the granular formulation along bed‑bug harborages, focusing on mattress seams, box‑spring crevices, and furniture joints. Apply 0.5 g per square meter; re‑treat after 14 days to target emerging nymphs.
Maintain records of temperature, humidity, and application dates. Regular laboratory checks of spore viability ensure consistent efficacy.