Snail and slug repellent as well as preparation method and application thereof
By using Bacillus amyloliquefaciens YN2010-P3 to prepare snail and slug repellents, the problems of inconvenience, inefficiency, and poor safety in existing methods for controlling snails and slugs have been solved. This method achieves efficient and safe biological control and has good market prospects.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-30
- Publication Date
- 2026-03-24
AI Technical Summary
Existing technologies for controlling snails and slugs are not simple enough, inefficient, and have poor safety, especially the use of chemical pesticides, which may cause environmental pollution and poisoning to humans and animals.
A repellent for snails and slugs was prepared using Bacillus amyloliquefaciens YN2010-P3. By spraying it on crops, the repellent's efficient reproduction and repellent effects can reduce the damage caused by snails and slugs to crops.
Snail and slug repellent has an effective period of at least 7 days, reproduces rapidly, has a high number of effective live bacteria, is stable in performance, is non-toxic to crops, has low production costs, is easy to use, and has good market development prospects.
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Figure CN121718474A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological prevention and control, and particularly relates to a snail and slug repellent as well as a preparation method and application thereof. BACKGROUND
[0002] Snails and slugs usually live in shrubs, low grass, farmland and dark and humid areas near houses, orchards and grassy slopes. They are nocturnal, and usually start to move and feed after dusk, reach the peak during 20-23 o'clock at night, and gradually reduce the amount of food after midnight, and stop feeding in the morning and hide in the soil or hide. They prefer dark and humid environments and can move and feed day and night on rainy days or after watering. Snails and slugs are omnivorous, and mainly eat leaves, stems, buds, flowers and juicy fruits of many crops, and usually move a lot in April-May and September-October, causing great harm to crops, and are important agricultural pests. The main measures to prevent and control snails and slugs at present include:
[0003] Agricultural prevention and control: After the weeds around the habitat of snails and slugs are cleaned and then decomposed into fertilizer, the environment suitable for snails is destroyed, and the source of snails is reduced. Cultivation loosens the soil, kills snails and slugs hiding in the soil, and destroys their hiding environment.
[0004] Physical prevention and control: Because snails and slugs will die of dehydration after contacting with lime, lime can be applied around the fields damaged by snails to prevent snails from entering the fields. According to the slow movement and nocturnal habits of snails and slugs, snails and slugs can be captured manually in the morning and evening or on rainy days. Weeds can be used to attract snails and slugs to feed, and then collected and eliminated the next day.
[0005] Chemical prevention and control: 0.5-0.6 kg of 6% Mida snail granules, 300 g of 30% polyaldehyde methanaphthol granules, 500 g of 10% tetraacetyl granules and other chemical pesticides are used per mu, and a proper amount of fine soil or fine sand is mixed, and then evenly applied in the field in the evening. This method has a certain control effect, but chemical pesticides may cause poisoning of humans and animals, and also cause environmental pollution.
[0006] As can be seen from the above, the common snail and slug control measures have certain control effect, but have the disadvantages of being not simple, low efficiency and poor safety, and therefore, it is necessary to develop a simple, efficient and safe biological prevention and control technology and preparation. SUMMARY
[0007] The purpose of this invention is to propose a snail and slug repellent, its preparation method, and its application for the control of snails and slugs. It has advantages such as simple nutrition, fast reproduction speed, high number of effective live bacteria, and stable performance. The control agent made from it is simpler, safer, and more efficient than traditional snail and slug control methods, and has good market development prospects.
[0008] The technical solution of this invention is implemented as follows:
[0009] This invention provides a Bacillus amyloliquefaciens YN2010-P3, whose taxonomic name is Bacillus amyloliquefaciens. The depository is the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is January 12, 2012, and the accession number is CGMCC NO. 5723.
[0010] This invention further protects the application of the above-mentioned Bacillus amyloliquefaciens YN2010-P3 in the control of snails and slugs.
[0011] This invention further protects a method for preparing a snail and slug repellent, wherein the above-mentioned Bacillus amyloliquefaciens YN2010-P3 is used to prepare a snail and slug repellent for the control of snails and slugs. The preparation of the snail and slug repellent includes the following steps:
[0012] (1) First, prepare liquid LB medium, sterilize it, cool it down, and obtain the medium for later use;
[0013] (2) Take a single colony of Bacillus amyloliquefaciens YN2010-P and inoculate it into the prepared culture medium obtained in step (1);
[0014] (3) The culture flasks obtained in step (2) are cultured to obtain Bacillus amyloliquefaciens YN2010-P3 fermentation broth, which is a repellent for snails and slugs.
[0015] As a further improvement of the present invention, the culture is carried out at a rotation speed of 170-180 r / min, a temperature of 35-40℃, a time of 44-52 h, and a culture medium pH of 6.5-7.5.
[0016] As a further improvement of the present invention, the bacterial concentration of the Bacillus amyloliquefaciens YN2010-P3 fermentation broth is 0.1-10×10⁻⁶. 8 CFU / mL.
[0017] This invention further protects a snail and slug repellent prepared by the above-described preparation method.
[0018] The present invention further protects a method for using the above-mentioned snail and slug repellent, wherein the snail and slug repellent is evenly sprayed onto bok choy.
[0019] The application of the snail and slug repellent as described in claim 6 in the control of snails and slugs.
[0020] As a further improvement of the present invention, the concentration of Bacillus amyloliquefaciens YN2010-P3, which repels snails and slugs, is 0.1-10 × 10⁻⁶. 8 CFU / mL.
[0021] As a further improvement of the present invention, the bacterial concentration of the *Bacillus amyloliquefaciens* YN2010-P3 is 1×10⁻⁶. 8 CFU / mL.
[0022] The present invention has the following beneficial effects:
[0023] This invention utilizes Bacillus amyloliquefaciens YN2010-P3 to prepare a snail and slug repellent with an effective period of at least 7 days. The snail and slug repellent of this invention is characterized by being pollution-free, having a rapid reproduction rate, a high number of effective live bacteria, simple nutrition, stable performance, non-toxic to crops, low production cost, and convenient to use; the product offers high cost-effectiveness and has good market development prospects. Attached Figure Description
[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0025] Figure 1 Colony morphology of Bacillus amyloliquefaciens YN2010-P3 on LB medium.
[0026] Figure 2 The attraction and avoidance index of Bacillus amyloliquefaciens YN2010-P3 to snails and slugs.
[0027] Figure 3 The area of worm holes in the control group (CK) after spraying with Bacillus amyloliquefaciens YN2010-P3.
[0028] Figure 4 The number of worm holes in the post-treatment and control group (CK) after spraying with Bacillus amyloliquefaciens YN2010-P3. Detailed Implementation
[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0030] The strain *Bacillus amyloliquefaciens* YN2010-P3 provided in this invention, taxonomically named *Bacillus amyloliquefaciens*, was deposited on January 12, 2012, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 5723. The address of CGMCC is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, postal code 100101. The strain was confirmed to be viable by CGMCC on January 12, 2012.
[0031] Example 1: Isolation and Screening of Strains YN2010-P3
[0032] Take 1 gram of wheat rhizosphere soil and add it to a 50 mL Erlenmeyer flask containing 9 mL of sterile water (10-fold dilution). Then, place the Erlenmeyer flask in a 60°C constant temperature sterilizer for 20 minutes. Remove the Erlenmeyer flask, place it on a clean bench, and shake well. Take 1 mL of soil solution into a test tube containing 9 mL of sterile water (100-fold dilution). Take 1 mL of soil solution from this test tube into a test tube containing 9 mL of sterile water (1000-fold dilution). Take 0.1 mL each of the 10-fold dilution, 100-fold dilution, and 1000-fold dilution, spread them evenly on LB solid agar plates, and incubate at 37°C for 48 h. Pick different single colonies and streak them onto new LB solid agar plates for purification. The purified bacterial strain was then inoculated into LB liquid medium and incubated at 37 ℃ with shaking for 48 h. The cultured bacterial solution was streaked onto fresh LB solid agar plates to check for purification. If the strain was purified, it was numbered and then incubated again in LB liquid medium, with 50 mL of LB medium added to each 100 mL Erlenmeyer flask. The flasks were incubated at 37 ℃ with shaking for 48 h. When the bacterial concentration reached 10⁸ CFU / mL, the bacterial solution was sprayed onto cabbage leaves and placed in a cardboard box (80 cm x 80 cm in diameter). Another cardboard box of the same size was added, along with cabbage leaves, and sprayed with water as a blank control. The boxes were sealed tightly with mesh. On the second and third days, the cabbage was observed for snail infestation holes. If the cabbage leaves sprayed with the bacterial solution have no snail-infested holes, while the water-based blank control has snail-infested holes, then the fermentation broth of this strain has a snail-repelling effect. If both the cabbage leaves sprayed with the bacterial solution and the water-based blank control have snail-infested holes, then the fermentation broth of this strain does not have a snail-repelling effect. Mix 40% sterile glycerol with the repellent bacterial solution at a 1:1 ratio and place the mixture in a sterile centrifuge tube. Store the mixture at -80°C for later use.
[0033] The bacterial strains isolated from dead snails were inoculated into LB liquid medium at pH 7 and fermented at 37℃ and 175 rpm for 48 h. The resulting fermentation broth was used to treat snails and slugs, respectively, to obtain a strain with high pathogenicity to snails and slugs, Bacillus amyloliquefaciens, strain code: YN2010-P3, which was stored in a -80℃ freezer with 40% glycerol.
[0034] Example 2
[0035] The application of Bacillus amyloliquefaciens strain YN2010-P3 in the control of snails and slugs involves preparing the Bacillus amyloliquefaciens strain YN2010-P3 into a snail and slug repellent for snail and slug control. The preparation method of the snail and slug repellent includes the following steps:
[0036] (1) First, prepare liquid LB medium, then dispense the prepared medium into Erlenmeyer flasks (500 mL), sterilize at 121 °C for 30 min, and then cool to 37 °C to obtain the ready-to-use medium.
[0037] (2) Take one single colony of Bacillus amyloliquefaciens YN2010-P3, i.e., 1 μL of bacterial cells, and inoculate it into the prepared culture medium;
[0038] (3) The obtained culture flasks were placed in a shaker at a speed of 175 r / min and a temperature of 37 ℃ for 48 h. The pH of the culture medium was 7. The resulting Bacillus amyloliquefaciens YN2010-P3 biocontrol agent was obtained with a bacterial count of 1×10⁻⁶. 8 CFU / mL.
[0039] Example 3
[0040] Evaluation method of indoor repulsion test of Bacillus amyloliquefaciens YN2010-P3 against snails and slugs:
[0041] Plant the bok choy in rectangular plastic trays, 12 plants per tray, with 6 plants spaced along the center line of the tray at a concentration of 1.0 × 10⁻⁶. 8 Six bok choy plants were treated with a CFU / mL bacterial solution (a repellent for snails and slugs) as a spray, while six plants were treated with sterile LB liquid medium as a control (CK). Both the control and treatment bok choy plants were air-dried before being planted in trays. The trays containing the planted bok choy were placed in 50×50×50 cm insect-rearing nets. Fifty snails and fifty slugs were placed at the boundary between the treatment and control areas, and the nets were closed to prevent escape. The nets were placed under suitable light and at room temperature, and the bok choy plants were watered regularly to keep the soil in the trays moist. Each treatment was replicated in triplicate. For 1-7 days after treatment, the following data were recorded daily for both the treatment and control areas: repellency index (RI), number of insect holes, and area of insect holes.
[0042] RI = (C−T) / (C+T)×100
[0043] In the formula: C is the number of snails or slugs appearing in the control area (heads); T is the number of snails or slugs appearing in the treatment area (heads).
[0044] The effects of spraying Bacillus amyloliquefaciens YN2010-P3 on the repulsion index of snails and slugs are evaluated as follows:
[0045] from Figure 2 It can be seen that YN2010-P3 has excellent repellency indices for both snails and slugs. One day after being sprayed on bok choy, the repellency indices for snails and slugs were 92% and 87%, respectively. On the second day, the repellency indices for both snails and slugs rose to 95%, but after the second day, the repellency indices for both snails and slugs began to decline. The repellency index for snails decreased from 95% on the second day to 82% on the sixth day, a decrease of 13%, and then rose again to 85% on the seventh day. The repellency index for slugs decreased from 95% on the second day to 87% on the fifth day, a decrease of 8%, then rose again to 88% on the sixth day, and then decreased to 81% on the seventh day.
[0046] The effects of spraying Bacillus amyloliquefaciens YN2010-P3 on the pore area evaluation of the control group are as follows:
[0047] from Figure 3 It can be seen that the worm hole area of the control (CK) was much larger than that of the treatment. The worm hole area of the CK was already significantly larger than that of the treatment on the first day, and on the second day, the worm hole area of the CK was 200 cm². 2 The area of the wormholes increased by 100+ cm² per day over the next 7 days. 2 The rate of increase was rising, and the area of the wormhole reached 892.27 cm² on the 7th day. 2 The treated wormhole area was only 37.41 cm² on day 7. 2 The wormhole area was 852.86 cm smaller than that of the control (CK) on day 7. 2 The efficacy of YN2010-P3 in controlling the disease in Chinese cabbage was 95.8%.
[0048] The effects of spraying Bacillus amyloliquefaciens YN2010-P3 on the number of worm holes in the control group (CK) are evaluated as follows:
[0049] from Figure 3 It can be seen that the number of worm holes in the control group (CK) was much higher than that in the treatment group. On the first day, the number of worm holes in the CK group was already higher than that in the treatment group, reaching 36. The number of worm holes increased most rapidly from the first day to the second day, increasing from 36 to 118. From the second day to the seventh day, the number of worm holes in the CK group increased by about 40 per day, reaching 330 on the seventh day. In contrast, the number of worm holes in the treatment group was only 35 on the seventh day, which was 295 fewer than the number of worm holes in the CK group on the seventh day.
[0050] Example 4
[0051] Evaluation method of field repulsion test of Bacillus amyloliquefaciens YN2010-P3 against snails and slugs:
[0052] The field repellency experiment was conducted on Chinese cabbage in the greenhouse of the organic vegetable planting base of Yunnan Yunling Fresh Agriculture Development Co., Ltd. Four Chinese cabbage plants of similar growth were selected from the greenhouse for the experiment. Two plants were sprayed with a bacterial solution (a repellent for snails and slugs) at a concentration of 1.0 × 10⁸ CFU / mL, while the other two plants served as a blank control. Seven days after spraying, both the treatment and control plants were sampled using a five-point sampling method, with nine Chinese cabbage plants taken from each point. The degree of damage was graded according to the proportion of insect holes on the leaves (Table 1). The damage index for the control and treatment plants was calculated based on the damage level, and the control efficacy was then calculated based on the damage index.
[0053] Table 1. Grading Standards for the Harmful Effects of Snails and Slugs on Bok Choy
[0054]
[0055] Harm Index = (∑Number of plants damaged at each level × Harm Level) × (Total Number of Plants Surveyed × Highest Harm Level) / 100 Control Efficacy (%) = (Control Harm Index − Treatment Harm Index) × Control Harm Index / 100
[0056] Field trial evaluation results:
[0057] Table 2 Hazard index and control efficacy in field avoidance trials
[0058]
[0059] As shown in Table 2, the damage index of Chinese cabbage sprayed with YN2010-P3 bacterial solution was 18.19±1.17 7 days after application, while the damage index of the blank control (CK) was 62.39±0.87. The damage index of the blank control was 44.2 higher than that of YN2010-P3. The field control efficacy of YN2010-P3 against snails and slugs was 70.84%.
[0060] The results above show that the concentration of the fungicide of the present invention, at 1.0 × 10⁸ CFU / mL, has a significant repellent effect on snails and slugs.
[0061] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A type of Bacillus amyloliquefaciens YN2010-P3, characterized in that, The taxonomic name of the Bacillus amyloliquefaciens YN2010-P3 is Bacillus amyloliquefaciens. The depository is the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is January 12, 2012, and the accession number is CGMCC NO. 5723.
2. The application of Bacillus amyloliquefaciens YN2010-P3 as described in claim 1 in the control of snails and slugs.
3. A method for preparing a snail and slug repellent, characterized in that, The *Bacillus amyloliquefaciens* YN2010-P3 described in claim 1 is used to prepare a snail and slug repellent for the control of snails and slugs. The preparation of the snail and slug repellent includes the following steps: (1) First, prepare liquid LB medium, sterilize it, cool it down, and obtain the medium for later use; (2) Take a single colony of Bacillus amyloliquefaciens YN2010-P and inoculate it into the prepared culture medium obtained in step (1); (3) The culture flasks obtained in step (2) are cultured to obtain Bacillus amyloliquefaciens YN2010-P3 fermentation broth, which is a repellent for snails and slugs.
4. The preparation method according to claim 3, characterized in that, The culture was carried out at a rotation speed of 170-180 r / min, a temperature of 35-40℃, a time of 44-52 h, and a culture medium pH of 6.5-7.
5.
5. The preparation method according to claim 3, characterized in that, The bacterial concentration of the Bacillus amyloliquefaciens YN2010-P3 fermentation broth was 0.1-10×10⁻⁶. 8 CFU / mL.
6. A snail and slug repellent prepared by the method according to any one of claims 3-5.
7. A method of using the snail and slug repellent as described in claim 6, characterized in that, Spray snail and slug repellent evenly onto the bok choy.
8. The application of the snail and slug repellent as described in claim 6 in the control of snails and slugs.
9. The application according to claim 8, characterized in that, The concentration of *Bacillus amyloliquefaciens* YN2010-P3, which repels snails and slugs, was 0.1-10 × 10⁻⁶. 8 CFU / mL.
10. The application according to claim 9, characterized in that, The bacterial concentration of the *Bacillus amyloliquefaciens* YN2010-P3 was 1×10⁻⁶. 8 CFU / mL.