GB/T 13917.1-2026Laboratory efficacy test methods and criteria for public health insecticides for pesticide registration - Part 1: Spray formulations (English PDF)

4.2 Test Target

4.2.1 Mosquitoes Culexpipienspallens (northern region) or Culexpipiens quinquefasciatus (southern region) (Region), sensitive strains of test insects were reared according to the method in NY/T 1964.2, and selected from the 3rd to 5th day after emergence, with a body weight of (2.0±0.2) mg. Unfed female adult worms.

4.2.2 Flies Houseflies (Musca domestica) were reared according to the method in NY/T 1964.1 for sensitive strains, and samples were selected on the 3rd day after emergence. On day 4, the adult insects weighed (20.0±2.0) mg/female and (16.0±1.6) mg/male, with half being female and half being male.

4.2.3 Cockroaches German cockroaches (Blattella germanica) were reared according to the method in NY/T 1964.4, and 10-day-old test insects were selected. Adult insects aged 20 days, weighing (100.0±10.0) mg/female (non-podded) and (50.0±5.0) mg/male, with half females and half males.

4.2.4 Ants The Yellow House Ant (Monomoriumpharaonis), worker ants raised in the laboratory.

4.2.5 Fleas Xenopsyllacheopis or Ctenocephalides felis, laboratory-bred fleas aged 3 to 10 days. Blood-sucking adult worms.

4.2.6 Bed bugs Temperate bed bug (Cimexlectularius), adult bed bugs raised in the laboratory, 3 to 5 days after feeding on blood.

6.3 Spray Test

6.3.1 Mosquitoes and flies A cylindrical apparatus (5.2) was used. The test insects (30 houseflies or 30 mosquitoes) were placed into the sealed cylinder through the central hole of the circular plate (E). Inside, tightly seal the rubber stopper (G). Prepare the test solution according to the recommended dosage of the test agent and the spraying volume specified in Table 1, and wait for the test insects to recover. After normal operation, transfer the test solution to the small guide tube using a micropipette or pipette. Vertically embed the nozzle (5.1) into the cylindrical device base. Secure the nozzle to the central hole of plate (F). Close the nozzle's air valve and connect it to the air compressor. Start the air compressor and... Adjust the pressure reducing valve to (9.8 × 10^4) Pa, and insert the flat-tipped No. 12 injection needle (D) under the nozzle into the bottom of the small tubing containing the test solution. Open the air valve and spray the pesticide. After spraying, tighten the rubber stopper (H), start timing immediately, and record the number of insects knocked down at regular intervals. After 20 minutes, the knocked-down test insects were collected into a clean rearing cage and restored to standard feeding conditions, fed with cotton balls containing 5% sugar water. Dead test insects were checked after 24 hours. Insect count. Insects not knocked down within 20 minutes should not be collected but counted as live insects. The test should be repeated three or more times. After the test, the insects should be cleaned. The entire experimental setup.

6.3.2 Cockroaches, ants, fleas, bedbugs A cylindrical device (5.2) was used, with a strip of Vaseline applied to the inner wall 30 mm from the bottom. Twenty cockroaches were released into 5.2; or ants... Fifty ants were released into a dish with a 90mm diameter plate containing a

1.1 suspension of 95% ethanol and talcum powder coated on the inner wall of the dish mouth. The dish was placed on a white filter paper base. Place the fleas in a petri dish and position the petri dish at the bottom center of 5.2; or release 50 fleas into a transparent cylindrical container with a bottom lined with white filter paper (5.7). In the middle, place the cylinder in the center of the bottom of 5.2; or release 20 bedbugs into a 90mm diameter petri dish with white filter paper at the bottom, and Place the petri dish in the center of the bottom of a 5.2-meter container. Prepare the test solution according to the recommended dosage of the test agent and the spray volume specified in Table 1. After the insects resume normal activity, transfer the test solution into the small tubing using a micropipette or pipette. Vertically insert the nozzle (5.1) into the colorless transparent... Secure the nozzle to the central hole of the acrylic disc (E). Close the nozzle's air valve and connect it to the air compressor. Start the air compressor. The compressor is adjusted to a pressure of (9.8 × 10^4) Pa via a pressure reducing valve. A flat-headed No. 12 injection needle (D) below the nozzle is then inserted into the container containing the test reagent. Open the air valve at the bottom of the small tube to spray the pesticide. After spraying, tighten the rubber stopper (G) and start timing immediately. For cockroach tests, record the time taken at regular intervals. Number of insects knocked down. After 20 minutes, all insects were collected in a clean container and returned to standard rearing. Cockroaches were fed mixed feed chunks and water-soaked cotton balls. Feed ants with water-soaked cotton balls; fleas and bedbugs do not need to be fed. Check for dead fleas, ants, and bedbugs after 24 hours (72 hours for cockroaches). The test should be repeated three or more times. The test apparatus should be cleaned after each test.

6.4 Residual spraying test

6.4.1 Preparation of the reagent contact plate surface Calculate the required dosage of active ingredient per unit area based on the recommended dosage of the test reagent for the contact surface, and adjust the dosage according to the water absorption of different plate surfaces. After diluting to the appropriate concentration, apply evenly dropwise or coat onto the contact plate surface, and allow to dry for later use. The coated plate surface should be naturally dried indoors. Store.

6.4.2 Determine the test time Test the coating every 15 days after spraying, until the specified testing time for the three types of panels (Table 3). Test within the specified time. If the mortality rate of a certain test insect on a certain board surface is less than 70% (mosquitoes, flies, fleas, ants, bedbugs 24h, cockroaches 72h), then that test insect is not harmful to that board surface. Continue testing.

6.4.3 Test Procedure

6.4.3.1 Mosquitoes, flies, and cockroaches Pull the baffle (C) of the forced contactor to the top. After lightly anesthetizing the test insects (20 mosquitoes, 20 houseflies, or 10 cockroaches), release them. The insect hole (H) is placed in the space between the forced contactor baffle (C) and the pull plate (D). After the test insect resumes normal activity, the forced contactor is placed... On the contact plate surface, pull out the pull plate (D) without harming the test insect, and at the same time push the pull rod (B) to push the baffle (C) to the bottom, forcing the test insect to... Immediately after contact with the treated surface, start timing. After 30 minutes, collect all test insects into a clean insect cage (mosquitoes, flies) or container (cockroaches) for recovery. For standard rearing, mosquitoes and flies are fed with cotton balls containing 5% sugar water, while cockroaches are fed with mixed feed blocks and water-soaked cotton balls. Mosquitoes and flies are checked for mortality after 24 hours (cockroaches after 72 hours). Number of test insects. The test should be repeated three times or more, with a blank control consisting of the same type of board surface that was not coated with the test agent.

6.4.3.2 Ants, fleas, bedbugs A bottomless transparent cylinder (5.6) was used. The transparent cylinder was placed vertically on the contact plate surface, and the test insect (ant) was placed in from above the transparent cylinder. Apply a ring of 95% ethanol and talcum powder (1.1) suspension to the inner wall of a cylinder, 1 cm from the contact surface, to 50 fleas; or 20 bedbugs. (Only one), immediately timed, 30 minutes, collect all test insects into a clean container, resume feeding, ants are fed with water-soaked cotton balls, fleas and bedbugs can be... Do not feed the insects; check the number of dead test worms after 24 hours. The test should be repeated three times or more, with a blank area of the same type of plate not coated with the test agent. Comparison.

7.Calculation The data from repeated tests were used to establish a toxicity regression equation using linear weighted regression, and KT50 was calculated. The results were then calculated using formulas (1) and (2). Mortality rates of mosquitoes, flies, fleas, ants, and bedbugs (72 hours for cockroaches) over 24 hours, and corrected mortality rates, rounded to two decimal places. The mortality rate of the blank control group was < 5%. No correction is required; if the mortality rate of the blank control is 5% to 20%, correction should be performed; if the mortality rate of the blank control is >20%, the experiment is invalid.