Mite killing activity evaluation method suitable for bee colony
Through the two-time mite killing treatment method, the number of bee mites that naturally fall off at the bottom of the beehive is counted, and the mortality rate of bee mites is calculated, which solves the problems of large error and non-intuitiveness of the bee mite parasitism rate detection method and realizes the accurate evaluation of acaricides.
Patent Information
- Application Number
- CN202510871007.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-26
- Publication Date
- 2025-10-17
AI Technical Summary
In the existing technology, the method for detecting the parasitism rate of bee mites is difficult to accurately reflect the killing activity of acaricides on bee mites in bee colonies, and has the problems of large errors and lack of intuitiveness.
The double mite killing method is adopted. By counting the number of bee mites that fall naturally at the bottom of the beehive, the mortality rate of bee mites is calculated, which directly reflects the lethal effect of the acaricide and avoids uneven distribution of bee mites and sampling bias.
The present invention provides a simple and easy method for evaluating acaricidal activity with high accuracy, which can dynamically evaluate the actual lethal effect of acaricides on bee mites and reduce the difficulty and error of operation.
Smart Images
Figure CN120806707A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of bee breeding and inspection and quarantine, and in particular to a method for evaluating acaricidal activity applicable to bee colonies. Background Art
[0002] Varroa mites, as the most important external parasite of honey bees, have become a major factor endangering the healthy development of the beekeeping industry, posing a major threat to the beekeeping industry and agricultural production that relies on bee pollination. Varroa destructor , commonly known as "Various mites") and bright heat mites ( Tropilaelaps Varroa mites (commonly known as "small Varroa mites") are parasitic pests caused by Varroa mites (commonly known as "small Varroa mites"). Varroa mites (large Varroa mites) suck honeybee hemolymph and transmit deadly viruses, severely weakening individual bee immunity and colony health, shortening bee lifespans, and ultimately leading to a decline in bee colony productivity, decline, or even extinction. Unlike plant and animal mites, over 90% of adult Varroa mites (large Varroa mites) lurk in the intersegmental membranes of the bee's ventral plate, while approximately 10% of adult Varroa mites (small Varroa mites) reside on honeycombs. Both the eggs and nymphs of both Varroa mites reside in uncovered or covered larval cells. Currently, treating bee colonies with acaricides is the primary technical means of effectively preventing and controlling Varroa mite disease. Therefore, effective evaluation of the acaricidal activity of acaricides is crucial for their development, efficacy assessment, and resistance detection.
[0003] In the prior art, the parasitism rate of bee mites is often used to evaluate the acaricidal activity of acaricides. The parasitism rate of bee mites includes the parasitism rate of bee mites in adult bees and the parasitism rate of bee mites in nest cells. The detection method for the former is: randomly select hundreds of bees in the bee colony, soak the bees with alcohol, detergent, etc., freeze the bees, or place the bees in powdered sugar and shake them to separate the bees from the bee mites, count the number of fallen mites, and calculate the parasitism rate of bee mites in adult bees by the ratio of the number of fallen mites to the number of sampled bees. The detection method for the latter is: select hundreds of capped nest cells containing larvae or pupae according to the season, check the number of bee mites, and calculate the parasitism rate of bee mites by the ratio of the number of bee mites to the number of sampled nest cells. However, the above two types of bee mite parasitism rates can only indirectly reflect the control effect of acaricides on bee mites in the bee colony, and it is difficult to intuitively and accurately characterize its killing activity against bee mites. At the same time, given the uneven distribution of bee mites in the bee colony (they prefer to parasitize on covered pupal cells or drone cells, etc.), the significant difference in the parasitism rate on the body surface of adult bees and the parasitism rate of unemerged individuals in the pupal cells, the small number of samples, insufficient statistical power, and the fact that they can only count the parasitism of bee mites at a certain stage, these traditional methods are not accurate and have large errors.
[0004] Therefore, developing a method for evaluating acaricidal activity suitable for bee colonies and accurately calculating the acaricidal activity of acaricides is of great significance for guiding the research and development of new acaricides, scientific prevention and control of bee mites, and improving the economic benefits of beekeeping. Summary of the Invention
[0005] In view of the prior art, the present application aims to provide a method for evaluating acaricidal activity suitable for a bee colony. The specific steps are as follows: cleaning the bottom of the bee colony box, counting the number of mites falling off after the first acaricidal treatment with the acaricide to be tested, then counting the number of mites falling off after the second acaricidal treatment with an acaricide having a different mechanism of action, and the sum of the number of mites falling off after the two acaricidal treatments is the total number of mites of the same mite in the bee colony, and the mortality rate of the mite after treatment with the acaricide to be tested is obtained by the ratio of the number of mites falling off after treatment and the total number of the same mite in the bee colony, which is used to characterize the acaricidal activity of the acaricide to be tested on different mites. The present application can dynamically evaluate the acaricidal activity of acaricides on mites, directly reflect the actual lethal effect of acaricides on mites, and effectively avoid errors caused by uneven distribution of mites and sampling bias.
[0006] To achieve the above object, the present application adopts the following technical solutions: The present application provides a method for evaluating acaricidal activity suitable for a bee colony, comprising the following steps: (1) After cleaning the bottom of the bee colony box, the first acaricidal treatment is performed on the bees in the bee colony box with the acaricide to be tested, and then the bottom of the bee colony box is cleaned again and the mites on the bottom of the bee colony box are collected, and the number of large mites A and the number of small mites B are counted respectively; (2) The second acaricidal treatment is performed on the bees in the bee colony box with an acaricide having a different mechanism of action from the acaricide to be tested, and then the mites on the bottom of the bee colony box are collected and the number of large mites C and the number of small mites D are counted respectively; (3) The mortality rates of large mites and small mites are calculated by formula (I) and formula (II) respectively to characterize the acaricidal activity of the acaricide to be tested; M 大蜂螨 (%) = [A / (A+C)] x 100% formula (I); M 小蜂螨 (%) = [B / (B+D)] x 100% formula (II); In the formula, M 大蜂螨 is the mortality rate of large mites, %; M 小蜂螨 is the mortality rate of small mites, %.
[0007] As a preferred, the operation of cleaning the bottom of the bee colony box in step (1) is as follows: the bee box is opened, the nest pulp in the bee box is taken out, and then the excess wax, wax chips, bee corpses and mites on the bottom of the bee colony box are cleaned with a scraper and a brush in turn, and then the taken-out nest pulp is put back in place, and the end of the nest door is raised to prevent the bees from cleaning the mite corpses out of the nest.
[0008] Further, the height of the raised end of the nest door is 2-4 cm.
[0009] As preferred, in step (1), the bottom of the beehive is cleaned again after 2-4 days after the treatment is finished.
[0010] As preferred, the method for counting the number of large mites and the number of small mites is as follows: the mites are observed and selected by using a magnifying glass or a stereoscopic microscope, and then classified and counted.
[0011] As preferred, in step (2), the mites are treated by using a mite-killing agent every 2-7 days during the second mite-killing treatment, and the mite-killing treatment is performed at least twice.
[0012] Further, the mites on the bottom of the beehive are collected 2-4 days after the last mite-killing treatment.
[0013] The beneficial effects of the present application are as follows: 1. The present application provides a mite-killing activity evaluation method suitable for a beehive, and the specific steps are as follows: the bottom of the beehive is cleaned, the number of mites falling off is counted after the first mite-killing treatment by using a mite-killing agent to be tested, the number of mites falling off is counted after the second mite-killing treatment by using a mite-killing agent with a different mechanism of action, the sum of the number of mites falling off after the two mite-killing treatments is the total number of mites of the same mite in the beehive, the mortality rate of the mite after the treatment by using the mite-killing agent to be tested is obtained by using the ratio of the number of mites falling off to the total number of mites of the same mite in the beehive, and the mite-killing activity of the mite-killing agent to be tested on different mites is characterized by the mortality rate, so as to determine the toxicity of the mite-killing agent to be tested on different mites.
[0014] 2. Different from the traditional detection of the parasitic rate of mites in a beehive, the present application is based on the characteristic that the mites naturally fall off the bottom of the beehive after death, the number of mites falling off the bottom is counted twice, the lethal effect of the mite-killing agent on the mites is dynamically evaluated, the actual lethal effect of the mite-killing agent on the mites is directly reflected, and the error caused by uneven distribution of the mites and sampling deviation can be effectively avoided. In addition, the mite-killing activity evaluation method of the present application is simple and easy to operate, the operation difficulty is reduced, and the detection result is accurate and reliable. BRIEF DESCRIPTION OF DRAWINGS
[0015] Figure 1 The flow chart of the mite-killing activity evaluation method suitable for a beehive in Example 1. DETAILED DESCRIPTION
[0016] It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the present application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
[0017] In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below with specific examples.
[0018] The experimental materials used in the examples of the present invention are all conventional experimental materials in the field and can be purchased through commercial channels. The experiments in the present invention were all conducted at the experimental bee farm of Shandong Agricultural University. The amitraz solution was produced by Shanxi Weipeng Pharmaceutical Co., Ltd. under the product name Weipeng Bee Mite Ke.
[0019] Example 1: Feasibility study of a method for evaluating acaricidal activity applicable to bee colonies Select 5 groups of Italian honey bees ( Apis mellifera ligustica ) bee colonies, each colony had about 5 full-frame bees for the experiment.
[0020] (1) Cleaning the bottom of the hive 101: Open the beehive, take out the honeycombs in the hive, and use a flat shovel and a brush to clean the wax, wax scraps, dead bees, and bee mites on the bottom of the hive. Then put the taken honeycombs back in place and raise the hive door by 3 cm to prevent the bees from cleaning the dead mites out of the hive.
[0021] (2) Experimental treatment 102: The first acaricide treatment was performed on the bees in the beehive box using fluvalinate. The specific operation was as follows: after mixing fluvalinate and water, a 0.02% acaricide solution was obtained. The acaricide solution was evenly sprayed on the surface of the honeycomb at a rate of 5 mL / comb. The acaricide solution was sprayed once every 3 days for 3 consecutive times. (3) Counting fallen mites 103: On the third day after the last spraying, open the beehive box, take out all the honeycombs in the box, and use a flat shovel and a brush to clean the wax, mites and bee corpses at the bottom of the box onto a clean plastic film (1m×1m). After cleaning away the bee corpses, large pieces of wax and other debris, collect the remaining bee mite corpses and other debris into a self-sealing bag and bring it back to the laboratory. Observe under the stereoscopic fiber optic cable, select the bee mites, and classify and count them; (4) Chemical acaricide 104: A second acaricide treatment was performed on the bees in the hive using a dimethoate solution with a different mechanism of action from the acaricide to be tested. The specific operation was as follows: a 0.001% acaricide solution was prepared by adding water to the dimethoate solution, and the solution was evenly sprayed on the surface of the honeycomb at a rate of 5 mL / comb. The solution was sprayed once every three days for three consecutive times. (5) Determination of Varroa Mite Parasitism Rate: Three days after the last spraying, 100 capped cells were randomly selected from the bee colony. The caps were opened with pointed tweezers. The surface of the caps or cells were carefully inspected for Varroa mites and Varroa mites. The number of mites that fell off was counted and the Varroa mite parasitism rate of the bee colony was calculated. The results are shown in Table 1. Varroa mite parasitism rate (%) = (number of mites that fell off / total number of bees inspected) × 100%.
[0022] Table 1 Results of the bee colony varroa mite parasitism rate inspection As can be seen from Table 1, after treatment with amitraz, which has a different mechanism of action from the test acaricide, the remaining bee mites in the bee colony can be basically killed, so the total number of mites that fell after the two acaricide treatments can be considered as the total number of mites in the bee colony. Based on this, this method can be used to evaluate the acaricidal activity of the bee colony, which is practical.
[0023] Example 2: An acaricidal activity evaluation method suitable for a bee colony On May 28, 2024, five Italian bee colonies (Apis mellifera) were selected (Colony 1, Colony 2, Colony 3, Colony 4, and Colony 5), each with a population of about 5 frames of bees and a parasitic rate of bee mites of ≥2%. Apis mellifera ligustica
[0024] (1) Clean the bottom of the box 101: On May 29, open the bee colony box, remove the brood comb from the bee colony box, and then use a flat shovel and a brush to clean the excess wax, wax chips, bee corpses, and bee mites on the bottom of the bee colony box. After cleaning, put the removed brood comb back in place, and raise the end of the nest door by 3 cm to prevent bees from cleaning the mite corpses out of the nest.
[0025] (2) Experimental treatment 102: The first acaricide treatment was performed on the bees in the bee colony box using flumethrin. Specifically, flumethrin and water were mixed to obtain a test acaricide solution with a volume fraction of 0.02%, and the test acaricide solution was evenly sprayed on the surface of the brood comb at a dosage of 5 mL / comb. The test acaricide solution was sprayed every 3 days, and the spraying was continued for 3 times. (3) Count the mites that fell 103: On the 3rd day after the last spraying (June 7), open the bee colony box and remove all the brood combs inside the nest box. Use a flat shovel and a brush to clean the excess wax, bee mites, and bee corpses on the bottom of the box onto a clean plastic film (1m x 1m). After removing the bee corpses, large pieces of excess wax, and other debris, collect the remaining bee mite corpses and other debris into a self-sealing bag and take them back to the laboratory for observation. Select and count the bee mites, and the results are as follows: The number of large bee mites that died in Colony 1-5 (A) was 192, 178, 169, 180, and 172, respectively. The number of small bee mites that died in Colony 1-5 (B) was 336, 325, 289, 301, and 278, respectively.
[0026] (4) Chemical acaricide 104: On June 8, a second acaricide treatment was performed on the bees in the bee colony box using amitraz, which has a different mechanism of action from the test acaricide. The specific operation is as follows: Amitraz solution was added to water to obtain an acaricide solution with a volume fraction of 0.001%, and the acaricide solution was evenly sprayed on the surface of the brood comb at a dosage of 5 mL / comb. The spraying was performed every 3 days, and the spraying was continued for 3 times.
[0027] (5) Secondary statistics of mites 105: On the 3rd day after the last spraying (June 17th), all the bee guts in the nest box were taken out, and the bottom wax, mites and bee corpses were cleaned to a clean plastic film (1m x 1m) with a flat shovel and a brush. After cleaning the bee corpses, large wax and other debris, the remaining mite corpses and other debris were collected into a self-sealing bag and taken back to the laboratory for observation, selection and counting of mites. The results are as follows: The number of large mites C that died in bee groups 1-5 was 45, 37, 32, 21, and 53, respectively; The number of small mites D that died in bee groups 1-5 was 51, 63, 42, 60, and 57, respectively.
[0028] (6) Mite-killing activity evaluation 106: The mortality rates of large mites and small mites were calculated by formulas (I) and (II), respectively, to represent the mite-killing activity of the test mite-killing agent. The results are shown in Table 1. M 大蜂螨 (%) = [A / (A+C)] x 100% Formula (I); M 小蜂螨 (%) = [B / (B+D)] x 100% Formula (II); In the formula, M 大蜂螨 is the mortality rate of large mites, %; M 小蜂螨 is the mortality rate of small mites, %.
[0029] Table 2 Killing effect of test mite-killing agent on large mites and small mites in bee groups As can be seen from Table 2, the mite-killing activity of the test mite-killing agent on large mites is 80.71%, and on small mites is 84.85%. The relative standard deviation (RSD) of the 5 bee groups shows that the RSD of large mites and small mites is less than 5%, proving that the test method is reproducible and reliable.
[0030] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.
Claims
1. A method for evaluating acaricidal activity of bee colonies, characterized in that: The following steps are involved: (1) After cleaning the bottom of the beehive, the bees in the beehive are treated with the acaricide to be tested for the first time. After the treatment, the bottom of the beehive is cleaned again and the mites on the bottom of the beehive are collected. The number of large varroa mites A and the number of small varroa mites B are counted respectively; (2) A second miticide treatment is performed on the bees in the beehive using an acaricide with a different mechanism of action from the acaricide to be tested. After the treatment is completed, the varroa mites at the bottom of the beehive are collected and the number of large varroa mites C and the number of small varroa mites D are counted respectively; (3) Calculating the mortality rates of Varroa spp. and Varroa spp. by formula (I) and formula (II), respectively, to characterize the acaricidal activity of the acaricide to be tested; M 大蜂螨 (%) = [A / (A + C)] × 100% Formula (I); M 小蜂螨 (%) = [B / (B+D)] × 100% Formula (II); Where M 大蜂螨 is the mortality rate of Varroa mites, %; M 小蜂螨 is the mortality rate of Varroa mites, %.
2. The method for evaluating acaricidal activity of bee colonies according to claim 1, wherein: In step (1), the operation of cleaning the bottom of the beehive is as follows: open the beehive, take out the honeycomb in the beehive, and use a scraper and a brush to clean the excess wax, wax scraps, bee corpses, and bee mites on the bottom of the beehive in turn, put the taken honeycomb back in place and raise the hive door end to prevent the bees from cleaning the mite corpses out of the hive.
3. The method for evaluating acaricidal activity of bee colonies according to claim 2, wherein: The height of the nest door end is 2-4cm.
4. The method for evaluating acaricidal activity of bee colonies according to claim 1, wherein: In step (1), the bottom of the beehive is cleaned again after 2-4 days after the treatment.
5. The method for evaluating acaricidal activity of bee colonies according to claim 1, wherein: In step (2), during the second acaricide treatment, acaricide is used to perform acaricide treatment every 2-7 days, and at least two acaricide treatments are performed.
6. The method for evaluating acaricidal activity of bee colonies according to claim 5, wherein: In step (2), the bee mites at the bottom of the beehive are collected 2-4 days after the last acaricide treatment.