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Tunnel Test for Mosquito Repellent Testing
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Summary
The tunnel test is a laboratory bioassay that can be used to assess the efficacy of mosquito repellents and insecticide-treated materials based on mosquito mortality and feeding success. This test imitates natural host-seeking behavior by allowing mosquitoes to migrate through the treated material towards a lure. The tunnel test is commonly used to test the efficacy of insecticide-treated bed nets, fabrics, and surface coatings.
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Evaluating the efficacy of repellent formulations, simulating real-world environments.
Quick understanding of the test
Tunnel Test for Insecticide treated Material - Evaluating the efficacy of insecticide-treated materials based on mosquito mortality and blood-feeding success
Application
- Anopheles
- Aedes
- Culex
- The assay evaluates the efficacy of mosquito-repellent treatments using a glass tunnel.
- This glass tunnel is sectioned off with a treated netting sample and bait to attract mosquitoes.
- 50 – 100 mosquitoes are released and their ability to pass through the treated material to reach the bait is measured.
- Post-exposure, mosquitoes are collected and their survival rates are assessed after 24 hours.
- Mimics real-world conditions to ensure precise evaluation of repellent effectiveness.
- Provides measurable data like repellency percentage and mortality rate.
Turnaround Time
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What Is the Tunnel Test for Mosquito Repellents?
The mosquito tunnel test is a controlled laboratory bioassay used to evaluate the biological activity of insecticide-treated materials under conditions that closely simulate host-seeking behavior. Unlike simple contact tests, this method forces mosquitoes to actively navigate through treated netting toward an animal bait, making the exposure more representative of real-world use conditions.
It is widely applied for mosquito nets and treated surfaces where blood-feeding inhibition, mortality, and behavioral effects must be demonstrated before field or semi-field evaluation.
Purpose of the Mosquito Tunnel Test
The primary purpose of the tunnel test is to determine whether a treated material produces a measurable biological effect on host-seeking mosquitoes. This includes assessing mosquito mortality, blood-feeding inhibition, and post-exposure effects such as reduced fertility.
The test is especially valuable for products designed to repel, irritate, or interfere with feeding behavior, rather than relying solely on rapid knockdown.
Mosquito Species Used in Tunnel Test
Mosquito species and strains are selected based on the intended mode of action of the product. Non-blood-fed mosquitoes (aged 5โ8 days) are required for the tunnel test.
Products Applicable for Tunnel Testing
Products tested under tunnel tests include insecticide-treated materials such as nets and fabrics.
Test Conditions and Exposure Duration
- Tunnel tests are conducted under strictly controlled environmental conditions, typically at 27 ยฑ 2ยฐC and 80 ยฑ 20% relative humidity.
- Exposure durations usually range from 12 to 15 hours, depending on the endpoint being measured.
- A suitable bait (such as a guinea pig or rabbit) is required to attract the mosquitoes
Tunnel Test Methodology Overview
- The assay is conducted in a 60-cm (25 cm x 25 cm square section) glass tunnel.
- A 25 cm square cage, covered with polyester netting, is attached at both ends of the tunnel.
- A bait (e.g., mouse) is placed in a small section of the tunnel to attract mosquitoes.
- A treated test sample is placed in a disposable cardboard frame at one-third of the tunnel’s length.
- Nine 1-cm holes are made in the netting sample to allow mosquitoes access to the bait.
- At the opposite end of the tunnel, 50โ100 mosquitoes are released towards the treated netting to see if they pass through to reach the bait.
- The mosquitoes from each section of the tunnel are collected and counted separately after 12-15 hours of exposure.
- An aspirator is used to collect the live mosquitoes, and the dead mosquitoes are removed from the bait chamber with the help of forceps.
- The live mosquitoes are transferred into plastic cups containing 10% sucrose solution and their mortality rate is recorded after 24 hours.
Parameters Measured in Tunnel Testing
Mosquito Mortality – It is the percentage of mosquitoes that die due to contact with the insecticide on the net. Mosquito mortality is recorded 24 hours after exposure to the treated net inside the tunnel.
Blood Feeding Inhibition – The ability of the treated net to prevent mosquitoes from successfully feeding on the blood source. After the overnight exposure period, mosquitoes are examined to determine whether they are blood-fed or unfed.
Reproductive Effects (Fertility and Fecundity) – For blood-fed mosquitoes, reproductive effects are assessed by observing egg development and egg-laying after a holding period.
Interpretation of Tunnel Test Results
Control group Mortality: Mortality in the control group should not exceed 10% after 24 hours. If it exceeds this, the test is invalid and must be repeated. For extended holding times, control mortality should not exceed 20%.
Blood Feeding in Control group : The blood-feeding success in the control group must exceed 50%. If it falls below this threshold, the test is invalid and should be repeated.
Precautionary Measures to Undertake while Testing
- Test conditions like temperature, humidity and lighting should be closely monitored and controlled.
- Testing personnel should wear protective equipment during testing to avoid accidental bites or sample contamination.
- Non-blood-fed, healthy mosquitoes must be used for the test. Mosquitoes must be handled with care to avoid any injury or stress, which can lead to variations in results.
- Animals used as baits must be handled ethically and humanely according to the approved guidelines.
- Strictly follow the exposure times to ensure consistency in results.
- Proper disposal of hazardous materials and dead mosquitoes must be followed according to the lab regulations for safety and environmental concerns.
Importance of the Tunnel Test for mosquito repellents
The tunnel test provides a controlled yet realistic assessment of the effectiveness of insecticide-treated materials against mosquitoes. The test is thus important to manufacturers to meet the required performance standards of the products. Additionally, testing ensures that repellent-treated materials inhibit the transmission of diseases like malaria and dengue by mosquitoes, thus contributing to public health.
Tunnel test vs Room test
Parameter | Tunnel Test | Room Test |
Purpose | To evaluate bio-efficacy of treated nets by measuring mortality and feeding inhibition | To evaluate mosquito repellency, knockdown, and mortality in an enclosed room |
Mandatory insects | Strains are selected based on the intended use of the product. | Aedes aegypti, Culex quinquefasciatus, Anopheles stephensi |
Scope of products | Insecticide-treated nets (ITNs) and fabrics and LLINs | Coils, vaporizers, mats, aerosols, spatial repellents, electric devices |
Test environment | Controlled laboratory tunnel system with bait source | Simulated real-use indoor room conditions |
Result interpretation | 24-hour mortality, blood-feeding inhibition, reproductive effects | Complete protection time (CPT), knockdown effect and mortality |
Why Choose Our Lab for Tunnel Testing
At MIS, we conduct Tunnel Test studies to assess the effectiveness of insecticide-treated nets and fabrics against mosquitoes in a controlled laboratory setting.
Our entomology laboratory, well-equipped with the necessary facilities, and our team of entomologists ensure the accuracy of study design, conduct, and interpretation.
Request a Tunnel Test Quote
To discuss your testing requirements and get a quote on tunnel test, contact our experts
Frequently Asked Questions
DR. Martinoz Scholtz
The tunnel test is a controlled laboratory bioassay used to evaluate the bio-efficacy of insecticide-treated materials, particularly mosquito nets.
Mosquito species for tunnel test are selected based on the intended use of the product.
Yes, mosquito nets are the primary products evaluated using the tunnel test.
Tunnel test measures 24-hour mosquito mortality, blood-feeding inhibition, and reproductive effects such as fertility or fecundity.
The tunnel test assesses mosquito behaviour and survival against treated materials using a bait source in a controlled system. In contrast, the arm-in-cage test directly measures repellency and bite prevention on human skin and is mainly used for topical repellents.
The key parameters measured during the tunnel test are the mortality rate of mosquitoes, the blood-feeding success rate, the repellency rate and the duration of repellency.
The tunnel test method is particularly suitable for insecticide-treated materials, such as bed nets, fabrics, curtains and wall coverings.
No, the tunnel test relies mostly on host-seeking adults since the test needs to establish the efficiency of repellents or insecticides against mosquitoes that usually bite and transmit diseases.
Multiple trials are carried out to ensure the reliability of the results. Generally, at least three independent trials should be carried out in controlled conditions to account for the variability in mosquito behavior and ensure statistical significance.
The repellency rate of mosquitoes is determined based on the percentage of mosquitoes deterred from passing through the treated material. This is done by comparing the number of mosquitoes that attempt to reach the bait in the treated and untreated sections of the tunnel.
In lab-controlled tunnel tests, the environment is highly uniform, and factors like temperature, humidity and exposure time are carefully controlled, while field tests provide insights into real-world performance.
The duration of repellent efficacy can be tested by conducting the test at different time intervals after treating the material with the repellent. The repellency and mortality rates over time can be measured to determine the duration of efficacy.
Tunnel testing is commonly required as part of WHO Prequalification and regulatory dossiers for insecticide-treated nets.
It takes 3-4 weeks to complete the Tunnel test.
In the tunnel test for mosquito repellents, mosquitoes are exposed to a repellent-treated material placed in a controlled tunnel environment. Mosquitoes are usually released in one section, with a bait at the opposite end. Mortality and blood-feeding success are measured.
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