Insectary

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Email: crid@crid-cam.com
Phone: +237 673 08 60 46

Our insectary comprises a total of nine rooms and one annex, organized as follows:

2 rooms for susceptibility testing to various insecticides
1 room for testing the bio-efficacy of ITNs (phase 1),
1 partitioned room dedicated to sensitive reference strains (FANG and NGousso Lab, FUMUZ and Kisumu),
1 lavarium and identification room,
1 room for Anopheles funestus rearing,
1 room for Anopheles gambiae rearing,
1 room for Aedes rearing,
1 room for other Anopheles and Culex species rearing,
Elende annex, for the mass production of susceptible strain and testing facilities (WHO cone bioassay, Tunnel test and the experimenta hut station).

All the rooms are under controlled and monitored conditions according to species rearing specificities, the system being maintained by air conditioning, humidifiers and heater system.

1-Biosafety

CRID established a rigorous system to prevent any escape of these insects into the external environment. These are some measures in place:

Double Entry Barrier:

The main entrance has a secure electronic door, forming a first physical barrier.
A second door, located a few meters from the first, constitutes an additional barrier. It is constructed with a glass panel and a durable mesh, providing double protection against escapes.
This double barrier creates a buffer zone, significantly reducing the risk of mosquitoes escaping when the doors are opened.

Internal Doors and windows Protection:

Doors and windows of the insectary rooms are protected by a durable mesh. This physical barrier prevents mosquitoes from leaving or entering rooms. The meshes also prevent mosquitoes from moving between the different areas of the insectary, avoiding strains contaminations.

Mosquito escape control:

Larvae: Hot water (90-100°C) is used to clean all rearing material to prevent the release of viable eggs or living larvae into drains.
Adults: Each rearing room is equipped with electrical devices designed to eliminate flying mosquitoes. These devices, such as UV light traps or insect destroyers, reduce the number of free adult mosquitoes in the rooms, thereby minimizing the risk of bites and escapes.

2-Bio-efficacy insecticide tests

Two primary bio-efficacy tests namely the WHO Bottle Bioassay and the WHO tube test, which are essential for monitoring and managing insecticide resistance in mosquitoes, are performed under laboratory controlled conditions. All the protocols follow the Standard Operating Procedure (SOP) for the WHO Tube test and WHO Bottle bioassays for evaluating the susceptibility of adult mosquito vectors to insecticides (WHO, 2022).

3-Bio-efficacy insecticide-treated nets tests

Various studies are conducted under both laboratory and field conditions to assess the efficacy and impact of different types of bed nets.

Phase 1: under laboratory conditions, bed-nets are tested against malaria vectors using the WHO cone test or Tunnel test to evaluate the efficacy and performance of insecticide-treated bed-nets (WHOPES, 2013). The WHO cone bioassay is used to assess the bio-efficacy of insecticide-treated nets (ITNs) under operational conditions while the tunnel test is used to evaluate the mortality rate and blood-feeding success of host-seeking mosquitoes.
Phase 2: in semi-field trials using Experimental Huts (EHT) to assess the efficacy of novel vector control tools, such as ITNS and Indoor Residual Spraying (IRS), against various malaria vectors. These semi-field trials provide valuable insights into the performance of vector control interventions under realistic but controlled conditions, helping to guide malaria prevention strategies.
Phase 3, conducted under field conditions to evaluate the ITNs durability and acceptability. These assessments focus on net survivorship (attrition), fabric integrity, and bio-efficacy over time.

4-Lab strains

All the reference strain available was summarized in Table 1 below.

Table 1: Susceptible strain information

Strain	FANG	FUMOZ	KISIMU	NGOUSSO	BENIN sus	BORA-BORA
Species	Anopheles funestus from Angola	Anopheles funestus from Mozambique	Anopheles gambiae from Kisumu, Kenya	Anopheles coluzzii from Yaoundé, Cameroon	Aedes aegypti from Benin	Aedes aegypti from Polynésie française
Resistance profile	Susceptible to all insecticidal classes	Resistant to all the insecticidal classes	Susceptible to Permethrin (100%), Alpha-cypermethrin DC (95.4%), DDT (81%), Bendiocarb (100%), Propoxur (100%), and Malathion (100%).	Susceptible to permethrin DC (86.4%), Malathion (100%), and resistant to DDT (18%).	Susceptible to Permethrin (100%), Deltamethrin (100%), Alpha-cypermethrin (100%), and bendiocarb (98%),	Susceptible to Permethrin (100%), Deltamethrin (100%), Alpha-cypermethrin (100%), Bendiocarb (90.17%), Pirimiphos-methyl (98,95%) and to Fenitrothion (100%).
Molecular characterisation	100 % S for CYP6P9a and CYP6P9b mutation	100 % R for CYP6P9a and CYP6P9b mutation	100% S for Kdr west mutation	100% S for Kdr west mutation

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