How do you cultivate Medilis for bed bugs? - briefly
Grow Medilis on a nutrient agar composed of 2 % glucose, 1 % yeast extract, and 1.5 % agar, incubating at 27 °C for 5–7 days until a dense mycelial mat forms. Harvest the spores, suspend them in a water‑oil emulsion, and apply the preparation to infested bedding, keeping humidity above 70 % to promote infection of bed‑bugs.
How do you cultivate Medilis for bed bugs? - in detail
To produce Medilis intended for Cimex control, follow a systematic protocol that addresses strain selection, growth medium, environmental parameters, inoculation technique, and post‑harvest handling.
Select a certified pure culture of the Medilis strain that exhibits proven efficacy against bed‑bug populations. Verify purity through microscopic examination and, if available, molecular confirmation.
Prepare a nutrient‑rich agar or liquid medium that supports rapid fungal proliferation. Typical composition includes:
- 20 g/L glucose
- 5 g/L yeast extract
- 15 g L⁻¹ peptone
- 0.5 g/L magnesium sulfate
- Adjust pH to 6.5 ± 0.2
Sterilize the medium at 121 °C for 15 minutes. After cooling to 45 °C, inoculate with 1 mL of a conidial suspension calibrated to 1 × 10⁶ conidia mL⁻¹. Mix gently to distribute spores uniformly.
Incubate the culture under controlled conditions:
- Temperature: 25–28 °C
- Relative humidity: 90–95 %
- Light: dark or low‑intensity fluorescent light, 12 h cycle optional
- Aeration: static for solid media; shaking at 150 rpm for liquid cultures
Maintain these parameters for 5–7 days, monitoring growth daily. Confluent mycelial mats or dense broth should develop, indicating readiness for harvest.
Harvest by scraping mycelium from solid plates or centrifuging liquid cultures at 3 000 g for 10 minutes. Resuspend the pellet in sterile distilled water containing 0.05 % Tween 80 to prevent clumping. Adjust final concentration to 1 × 10⁸ conidia mL⁻¹ using a hemocytometer.
Store the suspension at 4 °C for up to two weeks, protecting from light. For field application, mix the suspension with a carrier such as diatomaceous earth or a biodegradable polymer to enhance adherence to bedding surfaces.
Implement quality control checks before deployment:
- Verify spore viability (>90 %) by germination assay on agar plates.
- Confirm absence of contaminating microorganisms through plating on selective media.
- Record batch number, production date, and concentration on the label.
Follow local regulations regarding biological pest‑control agents, including required permits and safety data sheet provisions. Use personal protective equipment—gloves, goggles, and a lab coat—during handling to minimize exposure.