CN109362788B

CN109362788B - Granular mite killing agent and application thereof

Info

Publication number: CN109362788B
Application number: CN201811446098.8A
Authority: CN
Inventors: 胡文锋; 李楚君
Original assignee: Bioforte Biotechnology Shenzhen Co ltd
Current assignee: Bioforte Biotechnology Shenzhen Co ltd
Priority date: 2018-11-29
Filing date: 2018-11-29
Publication date: 2021-01-12
Anticipated expiration: 2038-11-29
Also published as: CN109362788A

Abstract

The invention provides a granular mite killing agent, which consists of one or more of microorganisms producing formic acid, propionic acid, lactic acid, acetic acid, butyric acid or valeric acid and acid substances produced by the fermentation of the microorganisms on a granular nutrient substrate. The organic acid contained in the granular mite removing agent comprises one or more of formic acid, propionic acid, lactic acid, acetic acid, butyric acid or valeric acid. The granular nutritive substrate comprises granular grains (rice, corn, wheat, sorghum, millet and the like) and soybeans and the like which are used as main nutritive substrates for fermentation. The granular fermented product has effect in repelling acarid. The granular substrate is used as a carrier of thalli and acid production through solid state fermentation, and can absorb moisture in the environment to continuously ferment and produce acid when in use, thereby continuously playing the role of expelling mites. The invention has good mite-killing effect, safety, harmlessness and no residue; the effective components are slowly released, and simultaneously, the fermentation can be continued to produce acid, so that the mite-expelling effect is enhanced and continued.

Description

Granular mite killing agent and application thereof

Technical Field

The invention belongs to the technical field of biological control, and particularly relates to a granular mite killing agent and application thereof.

Background

Mites are small animals belonging to the class of the arthropoda arachnids, the class of the Guangdong subclass. Phytophagous mites mainly comprise tetranychus, panonychus, goiter, rust mites and the like, are common pests, damage crops such as vegetables, fruit trees, cotton and the like, and often cause destructive disasters. Mites are more prevalent in animals, and almost every living location of animals, including humans, is infested with mites. In agriculture, the avian mites are the most common and seriously harmful ectoparasitic diseases in breeding of breeding hens (including ancestors and parents), laying hens and broilers (such as sanhuang chickens and capons) with the breeding period of more than 2 months, are distributed worldwide and are widely popular in laying hen farms in subtropical and temperate regions. In addition, rabbit sarcoptidosis, goat sarcoptidosis and pig sarcoptidosis are also extremely harmful in agricultural production. Insects are also harmful to mites, such as bee mites cause huge losses to the honeybee industry every year; in the breeding of hermetia illucens, mite infestation is seen especially in the breeding period of larvae.

The prevention and control of the mites mainly adopts three ways, namely, repelling the mites, killing the mites and blocking the invasion of the mites. Repelling mites are classified into smell and taste, and the smell repelling mites employ volatile substances such as pheromones, and the taste repelling mites employ organic acids. More mite-killing agents adopt the composite action of two methods. The common medicament of the chicken mites is amitraz, and the action mechanism of the amitraz is mainly to inhibit the activity of monoamine oxidase in mite bodies. The control of parasitic mites generally adopts methods such as heat treatment, trapping and chemical control, and the like, wherein the application of medicament control is the main measure for controlling parasitic mites at present. In production, the method for preventing and controlling mites generally combines the life habit of hosts, adopts methods of aqueous solution spraying, smoke fumigation, powder spraying, mite-killing hanging tablets and the like, and the most common medicaments comprise cyfluthrin, sublimed sulfur medicaments and the like.

The agriculturally important pest mites are particularly easy to generate drug resistance due to short breeding cycle, large breeding amount, small mobility and high selfing degree. Resistance frequently gives people a lot of difficulties in controlling mites. For this reason, new acaricides must be continuously developed. And the increase of the concentration of the acaricide threatens the food safety, also has negative influence on the environment and even endangers the food safety. It is therefore necessary to reduce the use of chemo-flooding/acaricides. However, the use of low concentrations of insecticides promotes an increase in pest resistance to the species; in addition, it is not guaranteed that all pests are killed and their growth cycle is destroyed. In order to avoid the defects, new technologies and new products which are safe, non-toxic, harmless, free of residue, sustainable and effective and are used for controlling mites need to be developed.

Disclosure of Invention

The granular mite killing agent provided by the invention has the advantages of good mite killing effect, low medication cost and safety and harmlessness to hosts. The mite killer utilizes the synergistic effect of the microorganisms, the compound organic acid generated by fermentation of the microorganisms and the auxiliary factors of the organic acid, plays a role in preventing and controlling mites, can continuously utilize the granular nutrient carrier to continue post-fermentation to generate acid while slowly releasing effective components, enhances the action effect and prolongs the service life.

The invention adopts the following technical scheme to realize the technical purpose:

a granular mite killing agent is obtained by adopting the following method: activating the microbial strain to obtain activated strain with a concentration of 10⁸～10¹¹cfu/ml, obtaining granular acidic leavening through solid state fermentation, and then drying at low temperature to obtain the mite removing agent; wherein the microbial strain is one or more of formic acid-producing bacteria, propionic acid bacteria, acetic acid bacteria, lactic acid bacteria or other acid-producing bacteria; preferentially, the methanogen is a necessary strain; other acid-producing bacteria are strains for producing butyric acid or valeric acid; the total acidity of the granular acidic fermentation product is 20 to 50g/100g (in terms of lactic acid).

The invention uses the solid state fermentation of the microorganism strains producing acid, preferentially uses the formic acid fermented by the methanogens and the like as the main active component, and simultaneously can be combined with other strains for fermentation together, and the produced organic acid and other substances are used as the mite removing component.

Preferably, the fermentation conditions for solid state fermentation are: soaking the granular base material overnight, adding auxiliary materials, then stewing at 120 ℃, sterilizing for 30 minutes, and curing, wherein the water content is 60-70%; when the temperature is cooled to 28-30 ℃, inoculating the activated strain, wherein the inoculation amount is 0.1-1% (v/w); fermenting for 24-48 hours at 30-40 ℃ under aerobic or anaerobic conditions, and drying at low temperature after fermentation.

The invention adopts a solid state fermentation method, for example, the grain particles of the invention are adopted for solid state fermentation, after low temperature drying, the fermented granular nutrient base material is simultaneously used as a carrier of microbial cells and organic acid, can absorb water when being used in an external environment, and can generate mite-removing substances after continuous after-fermentation, so that the effect is more effective and lasting.

Preferably, methanogens such as acetobacter woodii and the like; the propionic acid bacteria is one or more of acid producing propionibacterium, propionibacterium freudenreichii, propionibacterium scherzehnsonii, propionibacterium tehnsonii or propionibacterium beijing; the lactobacillus is one or more of Lactobacillus acidophilus, Lactobacillus casei, lactococcus lactis or Bifidobacterium lactis; clostridium butyricum producing butyric acid, and the like, but are not limited to the above-mentioned species and strains. Preferably, the microbial strains are selected to be safe to human, livestock and environment and even can be directly eaten.

Preferably, the solid-state fermented granular nutritional base material is beans or grains, such as one or more of soybean, soybean meal, rice, corn, sorghum, wheat or millet, and the auxiliary material is wheat bran and/or rice bran. Wherein, the base material accounts for 80 to 90 percent of the mass portion according to the mass portion; the auxiliary materials account for 10-20% by weight, and the initial pH of the fermentation base material is 5.0-7.2. The base material can be directly used as a solid fermentation nutrition main provider, and the auxiliary material is used as a growth factor provider, so that the fermentation can be better promoted.

Because the solid state fermentation method is adopted, the base materials, such as soybeans, bean pulp or corns, and the like, are controlled to be granular substances after being dried on the basis of the solid state fermentation, and the granular mite removing agent mainly plays the following modes:

1. the granular mite removing agent directly contains the effective components for removing mites after fermentation, and can directly release effective substances for expelling and removing mites;

2. because the nutrient substances contained in the particles can further grow and ferment microorganisms in the particles after absorbing water to generate organic acid, the mite removing effect is continuously exerted, and the mite removing effect is strengthened and prolonged;

3. the granular mite-killing agent can be directly eaten by livestock and poultry, is safe and harmless, and when the livestock and poultry excrete, the beneficial substances generated by the mite-killing agent through the fermentation of the intestinal tract still contain the effective components, so that the mite-killing and mite-killing effects in the growth environment of the livestock and poultry are facilitated.

Preferably, after fermentation is complete, drying is carried out at less than 60 ℃ to less than 10% moisture, either neat or under vacuum, while maintaining the granular shape.

The mite-killing agent is applied to livestock and poultry breeding, pet breeding and insect breeding environments and is applied to mite expelling in grain food or feed.

Further, the mite removing agent is applied to livestock and poultry, insect breeding environment or pet living environment, and the amount of the mite removing agent is 1-10 g per square meter; or mixed with grain and feed, and the granular mite removing agent is used according to the mass concentration of 0.1-10%.

Furthermore, the object to be treated by the granular miticide is harmful mites which are parasitic on the body surfaces of livestock, pets and insects, breeding environments and grains, grains and feeds.

The granular mite removing agent provided by the invention is directly applied to the ground of the livestock and poultry stall and the surrounding environment, such as the places where pest mites are easy to hide, the gaps of grasses and trees and the like, and can exert the effect in a larger range.

Compared with the prior art, the invention has the following advantages and beneficial effects:

the granular mite killing agent provided by the invention is prepared by fermenting acid-producing microorganisms which are safe to human, livestock and environment and even can be eaten, and has the advantages of high safety, good mite killing effect, good stability and lasting effect. The preparation process is simple to operate, the raw materials are easy to obtain, and the method is easy to popularize. The beneficial microorganisms are adopted to control mites, so that pesticide residue and harm to hosts can be effectively reduced, the drug resistance of pest mites is reduced, and meanwhile, the control cost and the pollution to the environment are reduced.

Drawings

FIG. 1 is a Y-shaped olfactometer experimental device used for mite killing test.

Detailed Description

The present invention will be further described with reference to the following specific examples and drawings, which are not intended to limit the invention in any manner. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the present invention are commercially available.

Example 1:

1. respectively activating commercially available acetobacter woodii, lactobacillus acidophilus, lactobacillus casei and propionibacterium freudenreichii, wherein the activated bacteria concentration is 10⁸cfu/ml, then mixing according to the mass ratio of 1:1:1:1, and inoculating the mixture into a granular fermentation medium as a fermentation strain.

2. Soaking soybeans and wheat overnight, draining, cooking at 120 ℃ for 30-50 minutes, discharging a heating material, uniformly mixing with wheat bran and cane sugar, and naturally cooling; wherein, the soybean and the wheat account for 90 parts by mass, the mass ratio of the soybean to the wheat is 1:1, the wheat bran accounts for 10 parts by mass, the sucrose accounts for 2-5%, and the total water content reaches 60-70%. Inoculating the activated strains in the last step according to an inoculation amount (v/w) of 0.5%, uniformly mixing, and carrying out anaerobic fermentation at 35-37 ℃ for 24-48 hours. And when the content of the organic acid is 35-50 g/100g, ending fermentation and drying to obtain granular acidic fermentation products containing the microbial cells and the organic acid, such as formic acid, lactic acid, acetic acid, propionic acid and the like.

3. Drying at below 60 deg.C under normal or vacuum condition to maintain the granule shape of the product, and oven drying to water content below 10% to obtain the final product.

Example 2:

1. activating commercially available Acetobacter woodii, Propionibacterium freudenreichii and Lactobacillus acidophilus respectively, wherein the activated bacteria concentration is 10⁸cfu/ml; mixing according to the mass ratio of 1:1:1, and using the mixture as a strain for solid state fermentation for later use.

2. Soaking the corn overnight, draining, and adding sucrose and bran coat in parts by weight of 7:0.5: 2.5; steaming and boiling for 50 minutes at 120 ℃ until the total water content reaches 60-70 percent; naturally cooling to 28-30 ℃, inoculating the activated strain in the last step according to the inoculation amount (v/w) of 0.2%, and uniformly mixing; carrying out micro aerobic-anaerobic alternate fermentation at 35-37 ℃ for 24-48 hours to obtain the organic acid content of 35-50 g/100g, and drying to obtain the granular fermentation product.

3. Drying the granular fermented product containing the microbial cells and the organic acid under vacuum at the temperature lower than 60 ℃, and drying until the water content is below 8% to obtain the acaricide.

Example 3:

1. activating commercially available Lactobacillus Woodwarfii, Lactobacillus acidophilus and Lactobacillus casei respectively, wherein the activated bacteria concentration is 10⁸cfu/ml; mixing according to the mass ratio of 1:1:1, and using the mixture as a fermentation strain for later use.

2. Soaking the wheat and the sorghum overnight, draining, and adding sucrose and bran coat, wherein the mass parts of the sucrose and the bran coat are 4:4:1:1 respectively; steaming and boiling the mixture for 30 minutes at the high temperature of 120 ℃ until the total water content reaches 60 to 70 percent; naturally cooling to 28-30 ℃, and then inoculating the activated strains in the previous step according to the inoculation amount (v/w) of 0.1%; carrying out anaerobic fermentation at 35-37 ℃ for 24-48 hours until the content of organic acid reaches 50g/100g, and finishing the fermentation; drying to obtain granular fermented product.

3. Drying the organic acid fermentation product at the temperature lower than 60 ℃ in vacuum, and drying until the water content is below 8% to obtain the mite-killing agent.

Application example 1:

refer to the experimental apparatus of Y-type olfactometer in FIG. 1. The granular mite removing agent in the embodiment 1 is weighed and prepared into a solution to be tested with the concentration of 0.1%, 0.3%, 0.5%, 0.8% and 1%, the solution is used as a medicine for an experimental group, filter paper is soaked, the solution is placed at the bottom of a taste source (bottle) 1, and mite feed (food) is placed above the filter paper and used as the experimental group. The other side of the flavor source (bottle) 2 is made by soaking filter paper with purified water and putting the same amount of mite feed as a control group. The air flow (right side of figure 1) is filtered by active carbon, and enters a flavor source bottle filled with the solution to be detected after being humidified by distilled water. And a gas flow velocity meter is connected before the odor source bottle is introduced to ensure that the flow velocity and the flow rate of the airflow at two ends are consistent. The gas passes through the odor source bottle to bring out the volatile gas components of the reagent to be tested, and the volatile gas components are converged at the middle part of the Y-shaped pipe. The chicken mites were released one by one at the left end of the Y-tube (left side of figure 1). The selection is made within 3min, the side of the experimental group stays for more than 1min and is recorded as 1, the side of the control group stays for more than 1min and is recorded as 0, the selection is not made, the tested acarid is taken out after each test, the inner wall of the Y-shaped tube is wiped by alcohol and is dried, the experiment is provided with 6 groups, 5 groups of experimental groups (respectively, filter paper soaked with the liquid to be tested with the concentration of 0.1%, 0.3%, 0.5%, 0.8% and 0.1%), 1 group of negative controls are arranged, and each 30 groups of negative controls are arranged. And (3) soaking filter paper in distilled water on both sides of the preliminary experiment, putting equivalent mite feed, and adjusting the position until the selection proportion has no obvious difference. During the experiment, the two sides of the Y-shaped olfactometer are ensured to be uniformly illuminated, and the positions of the flavor source bottles are exchanged once every 5 smell source bottles are measured during the experiment so as to eliminate the interference of the direction. The repellency rate was (1-mite number of experimental group/mite number of control group) × 100%. The results are shown in Table 1:

table 1: tendency of chicken mites

	Experimental group 1	Experimental group 2	Experimental group 3	Experimental group 4	Experimental group 5
						1 (only)	14	12	5	1	0
0 (only)	16	18	25	29	30
						Rate of repellency	10％	40％	80％	97％	100％

The experimental results are as follows: with the increase of the concentration of the mite killing agent, the situation that the mites tend to the control side is more and more, and the fact that the mite killing agent can have an obvious mite killing effect is proved, and the concentration is reliable.

Application example 2:

preparing the medicaments to be detected in a matching way: the miticide prepared in example 2 is mixed with water to prepare a liquid medicine to be tested with the concentration of 0.1%, 0.5%, 0.8% and 1%.

3 chicken farms with severe mite infestations were selected. 40 sick chickens are selected from the feed, averagely divided into four groups, raised in pens and naturally isolated.

Before medication, the old padding of the chicken coop is cleaned up, and the medication is started after 2 days when the new padding is replaced. Negative control group without any agent. Spraying the whole body of chicken with a small sprayer, gradually spreading the feather open, spraying the medicinal liquid to the deep part of feather and skin, and soaking feet in the medicinal liquid. And sprayed to the henhouse. The test period is about 30 days, the test conditions are observed and recorded every day, and the infection rate and the number of the focuses of each group are recorded by examination on the 1 st, 3 rd, 7 th, 16 th, 20 th, 25 th and 30 th days. The results are shown in Table 2:

table 2: drug effect and fruit record for mite-disease chicken

The experimental results are as follows: with the increase of the concentration of the medicament, the number of the focus is gradually reduced, and the infection rate is gradually reduced, which proves that the mite killing agent can obviously improve the diseases of the chicken mites.

Application example 3:

preparing the medicaments to be detected in a matching way: the granular fermented product of example 3 was mixed with water to prepare a liquid medicine to be tested having a concentration of 0.1%, 0.5%, 0.8%, 1%.

Selecting at least 3 hermetia illucens farms with serious mite infestation. 4 culture units (using pots as units) are randomly selected from each farm, are averagely divided into four groups, and are naturally isolated.

The 4 groups used the drugs to be tested (experimental groups 1-4), one group used the insect common acaricide monomethylamidine (control group), and one group used no any agent (blank group). Spraying the medicinal liquid for 3 times (1 time) every two days. Samples were taken from the culture unit 1 day before dosing and 7 days after dosing by five point sampling using a 5cm diameter cylinder. After soaking in 20% saline, mite numbers were recorded.

Table 3: mite damage heisui river horsefly administration effect record

The experimental results are as follows: along with the increase of the concentration of the mite killer, the mite killing rate is obviously improved, and the mite killer is proved to be capable of obviously improving the black soldier fly mite diseases.

Claims

1. The granular mite killing agent is characterized by being obtained by the following method: activating the microbial strain to obtain activated strain with a concentration of 10⁸～10¹¹cfu/ml, obtaining acidic granular fermentation products through solid state fermentation, and then drying at low temperature to obtain the mite removing agent; wherein the microorganism strain can be one or more of formic acid-producing bacteria, propionic acid-producing bacteria, acetic acid-producing bacteria, lactic acid bacteria or other acid-producing bacteriaA plurality of types; the formic acid producing bacteria are selected strains; other acid-producing bacteria are strains for producing butyric acid or valeric acid; the total acidity of the granular fermented product is 20 to 50g/100g or more in terms of lactic acid.

2. The granular miticide according to claim 1, wherein the fermentation process of solid state fermentation is: soaking the nutritional granular base material overnight, then adding auxiliary materials for cooking, sterilizing for 30 minutes at 120 ℃, cooling after curing until the water content is 60-75%; inoculating the activated strain, fermenting for 24-48 hours at 30-40 ℃ under aerobic or anaerobic conditions, finishing fermentation, and drying.

3. The particulate miticide of claim 1, wherein said formate-producing bacteria is acetobacter woodii; the propionic acid bacteria is one or more of propionibacterium acidovorans, propionibacterium freudenreichii, propionibacterium scherzehnsonii, propionibacterium tehnsonii or propionibacterium beijing; the lactobacillus is one or more of Lactobacillus acidophilus, Lactobacillus casei, lactococcus lactis or Bifidobacterium lactis; the butyric acid producing strain is clostridium butyricum.

4. The granular miticide according to claim 2, wherein the nutritive granular base material for solid state fermentation is one or more of soybean, soybean meal, rice, corn, sorghum, wheat or millet, and the auxiliary material is wheat bran and/or rice bran.

5. The granular acarus killing agent is characterized in that the nutritive granular base material accounts for 80-90% of the mass part; the auxiliary materials account for 10-20% by weight, and the initial pH value of the solid fermentation material is 5.0-7.2.

6. The granular miticide according to claim 2, wherein the inoculation amount of the activated strain is 0.1 to 1.0% (v/w).

7. The granular miticide according to claim 2, wherein after fermentation, the granular miticide is dried to a water content of 10% or less by ordinary or vacuum drying at a temperature of less than 60 ℃ to maintain the activity of the microbial cells and reduce the volatilization of all organic acids.

8. The use of the granular mite killing agent as claimed in any one of claims 1 to 7, wherein the granular mite killing agent is applied to livestock and poultry, an insect breeding environment or a living environment of pets, and the dosage of the granular mite killing agent is 1 to 10 grams per square meter; or mixing with grain and feed, wherein the mass concentration of the mite removing agent is 0.1-10% (w/w).

9. The use of the granular miticide according to claim 8, wherein the object to be treated by the miticide is a pest mite parasitic on the body surfaces of livestock, poultry, pets, insects, breeding environments, and in grains, foodstuffs, and feeds.