JPS5982085A - Biological control of vermin using useful microorganism - Google Patents

Abstract

PURPOSE:To achieve the vermin controlling effect and fertilizing effect, by culturing useful microorganisms in a medium composed of the waste originated from animal or vegetable, drying and granulating the proliferated microorganisms, and scattering the product to the soil or vegetables. CONSTITUTION:An animal- or vegetable-originated waste such as dung, human feces, bark, bean cake, rice straw, etc. is used as the cultivation medium, and aerobic mesophile, aerobic thermophile, anaerobic mesophile, anaerobic thermophile, or thermophilic actinomycete is cultured in the medium. The obtained fermentation product is allowed to cool, inoculated with various molds originated from fungi, and cultured. The cultured product is mixed with organic and inorganic materials such as 50% of zeolite and 1% of fish-meal, based on the cultured product. The mixture is dehydrated, and pulverized or granulated.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention uses waste materials such as human and livestock excrement residue as a medium to grow and proliferate useful microorganisms, and then dries and pulverizes the resulting material, which is then sprayed on soil or plants to eliminate pests and diseases. Concerning methods for obtaining pest control and fertilizer effects.

Conventionally, as seen in soil sterilization using T)-D agents, EDB agents, etc., many useful microorganisms are gradually decreasing due to chemical substances, resulting in a loss of soil vitality and a decline in productivity. This is a well-known fact, and in order to regenerate such soil, there are many cases where unripe organic fertilizers are applied to the soil, causing pests and diseases such as nematodes and mites to appear, repeating the vicious cycle. .

Therefore, in view of the current situation where organic farming as a pesticide-free farming method has recently been reconsidered, the present invention aims to overcome the drawbacks of the conventional methods described in 2. (1; J, T
"I-manufactured product". ) is used as a technical means to control pests and diseases, and at the same time provides an effective method for achieving the purpose of increasing soil fertility.

The present invention is based on the fact that approximately 4,000 strains of filamentous fungi and 500 strains of actinomycetes were isolated from nature, and while examining their killing effects on various pests, some filamentous fungi and actinomycetes showed their effectiveness. The present invention was conceived based on the above. In addition, similar results were obtained when strains with these killing abilities were identified and examined using known type cultures, and in the present invention, all experiments were conducted using type cultures.

The manufacturing method of this product will be explained below using Examples.

Primary cultures include animal-derived pig manure, cow manure, chicken manure, and human manure. Plant-derived products include park, rice bran, beet meal, soybean meal,
Some inorganic salts such as potassium nitrate, ammonium sulfate, various amino acids, peptone, etc. are added to tofu lees, rice straw, wheat straw, etc. The best combination of these is chicken manure, rice bran, tofu lees, and ammonium sulfate.
is adjusted to 7.0 to 7.2. In addition, those derived from the above-mentioned animals and plants may also be used. In the culture medium prepared in this manner, Bacillus subtilis (A
TCC6051), Bacillus megaterium (Bac
illus megaterium ATCC1458
]), Bacillus coagyulan 7. (Bacillus
s coagulans ATCC 7050), Micrococcuccus r') 7. (Mic-rococ
cus 1uteus ATCC398), and Bacillus stearothermophilus (Bacillus stearothermophilus) as a thermophilic bacterium.
llus s -tearothermophilu
s ATCC 12980), and Clostridium basteurianum (C-1o) as an anaerobic mesophilic bacterium (3).
stridium paste+anum AT
CC6013), Clostridium aminovalericum
mA, TCC13725), and Clostridium thermostrifolium (cl-ostri) as a thermophilic bacterium.
dium thermosaccharolytic
um ATCC7956), Thermoactinomyces vulgaris as a thermophilic actinomycete.
nomyces vulgaris ATCC14
570), Thermomonospora curvata (Therm
omonospora curvata KCC-A
-0042), and Pichia membranefaciens as yeast.
ens ATCC16046) ,) Torulopsis candida (Torulopsis candida AT
CC2560) was pre-cultured in large culture tanks with nutrient broth, malt extract broth, and peptone yeast broth, and anaerobic bacteria were pre-cultured in anaerobic culture jars, and equal amounts of each bacteria were mixed to incubate the animals and plants mentioned above. Static and aerated culture is carried out in the derived culture medium for about 2 weeks, during which temperature changes are carried out aerated for 1 to 3 days to raise the fermentation temperature to about 600 °C. Next 4
When cultured statically for ~14 days and allowed to grow anaerobic bacteria, 9
The temperature rises to the target of 5°C on the 10th day and falls to about 40°C on the 144th day. This temperature change kills all animal-derived intestinal bacteria and pests, leaving only useful bacteria alive.

Next, a filamentous fungus such as Aspergillus ostianus (Aspergillus ostianus We
hmerIFO4350), Aspergillus elegans (A, s-pergillus elegans)
Ga5perini IAM2136), Aspergillus ariaceus
allia -ceus Thom & Church
IAM 2159), Aspergillus saborivaaceus (Aspergillus 5ubol 1v)
ace-us, Raper & Fennell A
TCC16862) is subjected to secondary propagation using aerobic and anaerobic fermentation products derived from animals and plants.

Static culture is performed for about 5 days in a dark place at a humidity of 40 to 70% and a humidity of 20 to 308C. The growth status of the filamentous fungus can be determined by the growth of yellowish brown to green spores on the fermentation surface on the fifth day of culture.

- Mix ceolite and 1% fish meal at a ratio of 50% to the primary and secondary growth products, remove water, and form a powder or fine granules into a final product.

All the strains used above are from the American Strain Archives (A
merican Type Culture Co11
ction) and the Japan Federation of Microbial Conservation Institutions (Jl”C)
C) can be readily obtained from affiliated repositories.

As shown in Example 1, the safety of all strains used in this product according to the present invention has been confirmed. This is because, considering the specificity of the present invention as biological control, it is necessary to pay attention to the pathogenicity of the fungi introduced into the natural world to animals and plants.

Furthermore, as shown in Example 2, it was effective against a wide range of pests and diseases. Examples 3.4.5.6.7.8 in actual crates and in the field. However, as can be seen, the introduced bacteria do not show the same effect as pesticides in terms of soil colonization and control properties, but the product's control over time increases due to the growth of the bacteria in the soil. The effect is recognized. Another feature of this product is that it contains nutrients that allow useful microorganisms that control pests and diseases to rapidly proliferate in the soil. In other words, there were difficulties in biological control methods due to the effect of growth due to organic matter in the target soil, but this product overcomes this. On the other hand, as mentioned above, the method of soil sterilization reduces soil activity and causes problems such as pollution, and its effects are only temporary. In contrast, the present product does not have such defects and has useful bacterial groups of Examples 4 to 4.
8, the fact that concrete results have been confirmed due to the long-term pest control effect has produced a significant industrial effect compared to conventional technical ideas.

Next, the implementation procedure of the above-mentioned present manufacturing method is shown in the table below (Table-1).

(7) Example 1゜Two strains used in the present invention were used to examine whether they were pathogenic to animals or plants. First, mouse D mouse (female)
Diluted bacterial solutions and culture filtrates of each microorganism were intraperitoneally inoculated using 14 to 16 g (body weight: 14 to 16 g), and the number of deaths was observed for up to 20 days. Regarding chronic toxicity, cultured bacterial cells (including culture filtrate) were added to the feed, and the number of rearing days (deaths due to this) and pathological sections (due to this) were investigated.

(8) [Notes] 1) Number of bacteria 2) Number of deaths/number of survivors 3) Culture filtrate up to 3 ml No deaths were observed for each strain after 15 days of rearing.

4) Regarding chronic toxicity, each strain was
After 3 months of rearing (9//g), no deaths were observed, and no abnormal pathological findings were observed in the liver, kidneys, large intestine, and lungs.

5) No pathogenicity is observed for plants such as dice, Chinese cabbage, cabbage, potato, cucumber, and tomato.

Example 2゜Pot test on various pests and diseases Q: Controlling effect of this product,
house fly (adult), potato cyst nematode (adult), cabbage weevil (adult), cabbage weevil (adult)
Regarding the adult insects), the bacteria/filamentous fungi mixed fermentation product (including the culture filtrate) produced according to the present invention at a ratio of about 0.1% was added aseptically (along with the added feed) in a large petri dish, and the above-mentioned The number of dead insect pests was observed.Furthermore, the same secondary culture was used, and the filamentous fungi of the secondary culture were individually cultured and used in this test.

[Note]

1) Aspergillus ostianus (IF04350
)2) Aspergillus elegans (JAM 213
6) 3) Aspergillus saborivaaceus (ATCC
16862) 4) Aspergillus ant 7ceus (IA
M 2159) 5) Add this product to 1M house fly 3450ml milk solution at a concentration of 0.1%, and incubate in a mold Petri dish for 2 hours.
0 animals were released, and the number of deaths was measured after 4 days of rearing.

6) Add this product to 50g of Kokuzomun feed and steamed rice at a concentration of 0.1%.

Twenty animals were raised in a large petri dish, and the number of deaths after 4 days of rearing was measured.

7) 01% of this product was added to 50 g of feed mixed with starch and cabbage powder for locusts, and the number of deaths was measured after raising 50 locusts.

8) Add 0.1% of this product to 50g of artificial feed containing soybean powder, starch powder, glucose, sorbic acid, and phenoic acid.
The number of deaths after the addition of 50 animals for 5 days was measured.

9) 1% of this product was added to the artificial feed in 8), and the number of deaths was measured after 50 animals were fed for 6 days.

10) Adding 01% of this product to the artificial feed in 8), 50
The number of dead animals was measured after rearing the animals for 6 days.

Example 3 Crop damage control effect test (1) Test method A 1 m square wooden frame was buried in the field, filled with steam sterilization, and Aspergillus saborivaaceus (A
T CC16862), Anupergyranu ariaceus (I AM 21.59), Aspergillus
Ostianus (I FO4350), Anupergillus elegans (IAM2136) 108 pieces each/
After adding each amount of this product containing g, 100 cysts were inoculated, crops were planted, and an investigation was conducted.

Samples were collected by collecting rhizosphere soil and rhizomes every 30 days, and separating cysts and nematodes.

(2) Test outline a) The number of rests and eggs in the soil was measured by separating cysts from 50 g of air-dried soil using a sieve method (100 meshes), and the number of eggs was measured by crushing the cysts with a homogenizer.

b) The number of larvae and adult males in the soil is determined by
Pour into a container, stir while pouring water, pass through a 60-mesh sieve, pass through a 325-mesh sieve, and measure this using the Beer (1) Mann method.

C) Epidermis (Female adults attached to the soil were fixed with 5% formalin solution in 5g of fine roots and 1 tuber, brushed off with a brush, and measured using a 100-mesh sieve.

d) The number of adults in the root was measured by staining 1 g of the treated fine roots and 1 tuber with a phenol solution containing acidic fuchsin, and crushing with a homogenizer for 10 minutes immediately after release, and then separating and measuring.

e) Measurement of the number of live eggs of nematodes was performed using each field sample and the immersion experiment solution (3% maltose, 0.1% monopotassium phosphate, 0
．． Acridine orange solution diluted 2,500 times was added to 03% magnesium sulfate) and kept at 25°C for 48 hours.
Those that turn dark red are considered dead, and those that are unstained or lightly stained are considered semi-live.

f) Observation of crops affected by nematodes: Measure those that clearly show signs of nematode disease (comparison with control plots).

(12) Control effect test on potato cyst wires in a wooden frame As can be seen from the table above, there is no pesticide effect, but it can be said that it is a characteristic of biological control. , a high pesticidal effect is recognized.

Example 4゜ Field test. The test method is based on Example 3Q.

Application of this product in dry soil 100f in a continuous potato field A decrease in nematode density was observed over time.

Example 5 1. Prison test. The test method is based on Example 3.

009 This was a field application experiment of this product, and as in Example 4, the control effect of reducing nematode density was shown in Example 5, even in fields damaged by nematodes and nematodes. Recognized.

(15) Example 6 Nematode damage control effect of application of this product (Koyoru Potato 1) [Note] 1) To judge the avoidance effect, observe the rhizomes and growth inhibition of each planted plant, or observe the lines within the rhizomes. Depends on the number of insects.

In addition, in each test site, the amount of this product was applied at a rate of I kg per 10 ares, and the entire area was plowed after being applied.

As shown in the table above, when this product was applied to fields severely damaged by potato cyst nematodes, a remarkable control effect was observed, and it is recognized that it is useful in terms of practicality.

Example 'l.

(16) [Note] 1) Evaluation of effectiveness is based on Example 6.

Both cabbage test sites used this product at IK per 10 ares.
g was suspended in 1,000 times the volume of water, and the cabbage roots were immersed in one drop of the water and then planted.

At the Hakuzui test site, one product was sold to IJIK per 10 acres.
1. g with water. ．． A 1:000 solution was prepared and watered once over the entire surface.

As shown in Table 6, remarkable pesticidal effects were observed in the fields of Kiyokubetsu and Chinese cabbage.

Example 8゜Controlling effect of potato cyst nematode in a field damaged by potato cyst nematode 3 years after application of this product
US alliance) Period: From 1979 to 1982 Input: 10
Apply 1 kg of this product per area [Note] 1) Soil was collected from three points from each area of 25 acres, and after mixing, it was subjected to the measurement of the number of soils.

As shown in this table, it is recognized that the damage control effect coincides with the increase in the density of bacteria, and at the same time,
The strain used in this product has been found to have good colonization and multiplication properties in soil.

According to the method of the present invention, it is possible to realize an extremely effective means for promoting the development of reuse methods and the conversion of unused waste into secondary resources.In addition, even useful microorganisms are killed, unlike conventional soil sterilization. As a result, there is no need to worry about reducing the vitality of the soil, and there is no need to apply immature fertilizers that can lead to the spread of pests and diseases.In particular, a sustainable pest control effect can be obtained, and at the same time, it is possible to maintain and improve the fertility of the soil. It has the advantage of helping you achieve your goals.

Patent applicant Kou Atomic What Kasurishi Tadao Shirasawa

Claims (1)

[Claims] 1 Primary culture using aerobic mesophilic bacteria, anaerobic mesophilic bacteria, and thermophilic actinomycetes using animal and plant-derived waste as a culture medium, and then propagating various fungal-derived filamentous fungi. A biological control method for controlling pests and diseases using a group of useful microorganisms, which is characterized in that it is mixed with an inorganic substance, water removed, pulverized, and applied to soil and plants.