CN106399149A - Medium formula of mosquito larva killing preparation, namely, wiggler killer - Google Patents

Medium formula of mosquito larva killing preparation, namely, wiggler killer Download PDF

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Publication number
CN106399149A
CN106399149A CN201610528775.5A CN201610528775A CN106399149A CN 106399149 A CN106399149 A CN 106399149A CN 201610528775 A CN201610528775 A CN 201610528775A CN 106399149 A CN106399149 A CN 106399149A
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China
Prior art keywords
mosquito
killing
bacillus
mosquito larvae
larvae
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CN201610528775.5A
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Chinese (zh)
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徐军
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Bazhou Point Of Green Agriculture And Forestry Technology Co Ltd
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Bazhou Point Of Green Agriculture And Forestry Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/10—Animals; Substances produced thereby or obtained therefrom

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Virology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Environmental Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The invention provides a mosquito larva killing preparation, namely, wiggler killer, which is colorless and odorless. The mosquito larva killing preparation is specially used for killing mosquito larvae, namely, wigglers, the larva killing is broad-spectrum, the effects are stable, the environment is not polluted, and the mosquito larva killing preparation is non-toxic and harmless to human beings, beasts, birds, fish and shrimps, and beneficial insects, and can completely purify the water quality, so that the comprehensive prevention and treatment effects are achieved. The bacillus is separated from the mosquito larva falling ill naturally, and grows well in an LB culture medium. Through separation and purification, and physiological and biochemical identification, the bacillus is bacillusthuringiensis subsp israelensis Bti, and the bacillus has extremely strong lethality for eight mosquito larvae including culex pipiens pallens. Toxin protein produced during the growth of the bacillus plays the killing role, and after eating by the mosquito larvae, the pepsase of the mosquito larvae is destroyed within 2-12 h, so that the mosquito larvae can not grow and develop, and die of sepsis. During the spore forming process, Bti synthesizes five active proteins capable of killing the mosquito larvae.

Description

Mosquitocidal children's clear culture medium prescription of preparation wriggler
Technical field
The present invention be through separating, screening, a kind of pathogenic microorganism that repeatedly purifying, physiology, biochemical identification obtain thin Bacterium, this bacterium is Bacillus thuringiensissubsp.israelensis（Bacillusthuingiensis subsp israelensis is referred to as Bti).This bacterium has been cooked a large amount of separation, screening, purifying, physiology, biochemical identification, 16SDNA sequence analysis and multiple dynamic in laboratory Outside the safety testing such as chronic, subacute, acute of thing, also carry out shaking flask, canister, middle filling, big tank and 10 tons and fill producing fermentation. In addition to being capable of stablizing industrialization production it is often more important that, also to filter out can suitably Bti Fermentative growth well required The carbon source wanted, nitrogen source, phosphorus source culture medium active ingredient.Our company, in long-term production fermentation, have developed out suitable Bti fermentation life Length is vigorous, Number of spores is many, gemma quality is good, high-quality forms crystalline toxin（This Bti bacterial strain is during Fermentative growth More than 5 kinds crystalline toxins can be produced）Culture medium.Bacillus thuringiensis subsp israelensis (Bti) are a kind of important Inclusive type pathogenic microorganism bacterium, it is known that 40 serum kinds and 54 serum subspecies can be divided into, has toxic effect to mosquito larvae wriggler Bacterial strain have 3 kinds, serotype be H-14 (Israel subclass), PG-14 (Mo Shi mutation) and 73-E10-2 (Darmstadt change Kind), most widely used is bacillus thuringiensis subsp israelensis.BtHi-14 is by Goldbery (1972) and Margelit (1977) isolate in Israel.This bacterium has higher selectivity to mosquito larvae wriggler, and in host intestine, crystalline protein is former Toxin, in high ph-values and in the presence of the enzyme that is suitable for, is transitted very quickly into less poisonous subunit, mosquito larvae wriggler eats crystalline substance After body protein, cause midgut epithelial cellses to rupture sepsis illness and dead.China 20 century 70 starts Israel subclass is entered Row scale up test, cesspool, rice terrace are applied with concentration 1 ~ 3mg/L in the wild for mosquitocidal children's agent of production, kill culex within 24 hours, 90% is reached to albopictus larvae wriggler prevention effect, also has good result to Anopheles Sinensis Larvae wriggler.So far said preparation annual output is hundreds of Ten thousand tons, it is mainly used to kill the albopictus larvae wriggler in container ponding, particularly the Aedes aegypti larva in drinking-water water vat Wriggler.There is the application of larger area in China south and north, after dispenser 1 ~ 2 day, mosquito larvae wriggler density is decreased obviously, efficient All 93% ~ 95.3%.In the bacillus thuringiensis subsp israelensis 187 floating particle agent preventing and treating rice terrace of domestic development Anopheles sinensis, the lasting period is 16 days；The mosquitocidal children preparation wriggler that Ba Zhou Dian Lv agriculture and forestry science and technology Co., Ltd develops is clear in the wild After the ground dispensers such as reed wetland, sewage treatment plant, rice terrace, sewage of farm discharge areas 1 day, come to an end fruit, kills light color storehouse Mosquito larvae wriggler 98%, kill Anopheles Sinensis Larvae in Paddy Fields wriggler 100%, kill Culex Quinquefasciatus Larvae wriggler 100%, kill act of violence Little Ability of Culex Pipiens Larvae wriggler 99%, kill graceful mosquito larvae wriggler 100%, kill and mosquito larvae wriggler 98%, some regions 100%.Lasting period all more than 45 days, effect stability.
Background technology
The clear bacterium using of mosquitocidal children preparation-wriggler is the bacillus thuringiensis strain Israel subclass being separated to；Belong to The long bacillus of Gram-positive, under the conditions of suitable pH value and nutritional sufficiency, bacillus, in growth and development process, can form ellipse Shape spore.Bacillus, when growing formation spore final stage, is grown to serve as the proteotoxin-egg with extremely strong lethality Bai Jingti.After mosquito larvae wriggler takes food albumin crystal entrance alimentary canal, the δ-endogenous toxic material compared with subsection is broken to by alkaline intestinal juice Element, destroys the pepsin of mosquito larvae wriggler, makes middle intestines stop wriggling, midgut epithelial cell slowly dissociates, and mosquito larvae is lonely Jue starts stopping and takes food.Gemma starts to sprout in the middle intestines of mosquito larvae wriggler, enters through destroyed intestinal wall after haemocoele and starts Amount reproduction, finally causes mosquito larvae wriggler total paralysis, makes mosquito larvae wriggler obtain sepsis illness and dead.
Mosquitocidal children's clear product function of preparation wriggler：Mosquitocidal larva wriggler wide spectrum, effect stability, lasting period are long, to people, Poultry, fowl and beneficial insect are nontoxic, harmless, safe, the easy to use, saving of work and time of application, free from environmental pollution, convenient transportation, target mesh Standard is true, stable performance.It is to kill mosquito larvae wriggler, effective control and reduce into mosquito density, adult mosquito propagated filariasis, step on Leather heat, malaria, yellow fever, encephalitis, onchocercosis, propagation of arboviruse infectious disease etc. is preferably minimized limit.Long-Time Service, Do not produce resistance moreover it is possible to thoroughly purify water.
Mosquitocidal children's clear mechanism of action of preparation wriggler：Main active is half peritectoid-proteotoxin.Mosquito larvae- The pepsin of wriggler after wriggler takes food, can be destroyed quickly, the delta-endotoxin compared with subsection is broken to by alkaline intestinal juice.In making Intestines stop wriggling, and midgut epithelial cell slowly dissociates, and mosquito larvae wriggler starts stopping and takes food.Gemma is in mosquito larvae wriggler Intestines are sprouted, and enter through destroyed intestinal wall after haemocoele and start amount reproduction, cause mosquito larvae wriggler total paralysis, make mosquito children Worm wriggler, impotentia takes food and obtains the death of sepsis illness.
Mosquitocidal children's clear consumption of preparation wriggler：Waters area is 1-3ml/m2.Poultry, cattle breeding field, pig farm, cowboying Field, sewage treatment plant, city grid area, municipal sewage system, river, the serious area of weir Tang Deng region mosquito larvae wriggler generating capacity Domain or the high region of the content of organic matter, are 3-5 ml/m2.
Content of the invention：Present invention is mainly used for the liquid fermentation medium of mosquitocidal children's clear industrialization production of preparation wriggler is joined Side.
Culture medium prescription (%)（LB slant medium global general-use omits herein）
Fermentation broth：
Dusty yeast 3.0
Corn steep liquor 2.5
Glucose 2.0
Calcium chloride 0.1（CaCl）
Calcium carbonate 0.1（CaCO3）
Potassium dihydrogen phosphate 0.1（KH2P04）
Additive 0.3 B B-complex is some
PH value (before sterilizing) 7.5
Carbon source, phosphorus source culture medium：
Carbon source 2.5
Potassium dihydrogen phosphate 0.3（KH2PO4）
Calcium chloride 0.2（CaCl2）
Zinc chloride 0.03（ZnCl2）
Ferric trichloride 0.05（FeCl3）
Magnesium chloride 0.005（MgC12）
Cobalt chloride 0.005（CoC12）
Copper chloride 0.005（CuC12）
Additive 0.3
B B-complex is some；
Nitrogen source, phosphorus source culture medium：
Nitrogen source 2.7
Potassium dihydrogen phosphate 0.3（KH2PO4）
Dipotassium hydrogen phosphate 0.3（K2HPO4）
Calcium chloride 0.2（CaC12）
Zinc chloride 0.05（ZnC12）
Ferric trichloride 0.05（FeC13）
Cobalt chloride 0.005（CoC12）
Copper chloride 0.005（CuC12）
Additive 0.3
B B-complex is some：
Single carbon source, nitrogen source recipe determination raw material：
Sweet potato powder, dehydrated potato powder, tapioca starch, corn-dodger powder, corn flour, molasses.
Single nitrogen source recipe determination raw material
Peptone, dusty yeast, soybean cake powder, dried silkworm chrysalis meal, fish meal, groundnut meal, cottonseed meal（Must not Insect Resistant Cotton cottonseed cake）.
Bacillus thuringiensis subsp israelensis chitinase amino acid sequence
Ichi 151 DCDKYARCGNFGELKRLKAKYPHLKTIISVGGWTWSNRFSDMAADEKTRKVFAFSTVAFL
Ac 243 SAWDEPYKGNFGNLMALKQAHPDLKILPSVGGWTLSDPFYFLG-DKTKRDTFVASVREFL
Al 241 VDAKDPIRGVYSQLMALKQRYPDLKILPSVGGWTLSDPFHGFT-NKANPDTFVASVKQFL
BcAl 132 DTWDQPIAGNINQLNKLKQTNPNLKTIISVGGWTWSNRFSDVAATAATREVFANSAVDFL
Bl 128 DVWDEPIRGNFKQLLKLKKSHPHLKTFISVGGWTWSNRFSDVAADPVARGNFAASAVEFL
Bs 125 DTWDEPIRGNFKQLLKLKKNHPHLKTFISVGGWSWSNRFSDVAADPAARENFAASAVNFL
Ct 91 AILDTPIRGHFGQLIKYKEQYPHVRTLISVGGWTESKYFSDVALTEESRNTFADSCVEFL
Sm 243 TAWDDPYKGNFGQLMALKQAHPDLKILPSIGGWTLSDPFFFMG-DKVKRDRFVGSVKEFL
Ichi 211 RAYG-FDGVDLDWEYPGVETIPGGSYRP-EDKQNFTLLLQDVRNALN
Ac 302 QTWKFFDGVDIDWEFPGGQGANPSLGGP-NDGATYVVLMKELRAMLD
Al 300 KTWKFYDGVDIDWEFPGGDGPNPDLGDPINDGPAYVALMQELRAMLD
BcAl 192 RKYN-FDGVDLDWEYPVSGGLDGNSKRP-EDKQNYTLLLSKIREKLD
Bl 188 RKYG-FDGVDLDWEYPVSGGLPGNSTRP-EDKFNYTLLLQEVRKKLD
Bs 185 RKYG-FDGVDLDWEYPVSGGLPGNSTRP-EDKFNYTLLLQDVREKLD
Ct 151 RTYR-FDGVDIDWEYPVSGGMPENIRRP-EDKQNFTLLLKCLREKLD
Sm 302 QTWKFFDGVDIDWEFPGGKGANPNLGSP-QDGETYVLLMKELRAMLD

Claims (1)

1. bacillus thuringiensis subsp israelensis are widely present in nature, the research institution of external and China and part junior college Universities and colleges, successively carry out different degrees of research and application to this bacterium, and therefore, each personal medium component is different；Due to Medium component is different, and quantity, the Yield and quality such as the gemma of fermentation cultivation, crystalline toxin all can have great difference；And The culture medium prescription of the culture medium prescription that we develop, particularly liquid fermentation production and all different, the institute of other unit The potency of the bacillus Israel subclass preparation produced with us, it is many to exceed other manufacturer's twices, country's mark Standard is 400 units, and other producers all can only achieve 350 units, Ba Zhou Dian Lv agriculture and forestry science and technology Co., Ltd develop all 1000 More than unit（The present invention, in 10 tons of fermentation tanks, is fermenting substrate 28 hours with soybean cake powder, cornstarch etc., sampling detection, Potency average out to 2899Iu/uL, its performance indications respectively reaches 22100Iu/mg, viable spore number 4*1709/more than g, carefully Spend and pass through standard 300 mesh sieve for 99%）, so, special requirement obtains having and must protect to the formula of this medium component.
CN201610528775.5A 2016-07-05 2016-07-05 Medium formula of mosquito larva killing preparation, namely, wiggler killer Pending CN106399149A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109593680A (en) * 2018-12-26 2019-04-09 武汉科诺生物科技股份有限公司 A kind of Dipel liquid fermentation medium and its bacterium powder and oil suspending agent

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5202240A (en) * 1985-11-27 1993-04-13 Ciba-Geigy Corporation Preparation of spore free, concentrated protein preparations from Bacillus thuringiensis serovar,israelensis, which is toxic
CN1368549A (en) * 2002-03-06 2002-09-11 中国科学院武汉病毒研究所 Tharingiensis bacillus strain with broad-spectrum insecticiding activity and its preparing process
CN1915030A (en) * 2006-09-07 2007-02-21 广东省生态环境与土壤研究所 Anopheline of microbe prepared from starch wastewater, and preparation method
JP2008048682A (en) * 2006-08-25 2008-03-06 Fukuoka Prefecture Bacteria belonging to genus bacillus and use of the same
CN102250818A (en) * 2011-07-23 2011-11-23 福建农林大学 Method for preparing solid fermentation culture medium for bacillus thuringiensis
CN102286413A (en) * 2011-09-03 2011-12-21 福建农林大学 Preparation method of liquid fermentation medium for bacillus thuringiensis
WO2013005176A1 (en) * 2011-07-05 2013-01-10 Yeditepe Universitesi Novel bacterial strains for biological control of mosquitoes
CN102907458A (en) * 2012-10-28 2013-02-06 郑文定 Biological mosquitocide and preparation method thereof

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5202240A (en) * 1985-11-27 1993-04-13 Ciba-Geigy Corporation Preparation of spore free, concentrated protein preparations from Bacillus thuringiensis serovar,israelensis, which is toxic
CN1368549A (en) * 2002-03-06 2002-09-11 中国科学院武汉病毒研究所 Tharingiensis bacillus strain with broad-spectrum insecticiding activity and its preparing process
JP2008048682A (en) * 2006-08-25 2008-03-06 Fukuoka Prefecture Bacteria belonging to genus bacillus and use of the same
CN1915030A (en) * 2006-09-07 2007-02-21 广东省生态环境与土壤研究所 Anopheline of microbe prepared from starch wastewater, and preparation method
WO2013005176A1 (en) * 2011-07-05 2013-01-10 Yeditepe Universitesi Novel bacterial strains for biological control of mosquitoes
CN102250818A (en) * 2011-07-23 2011-11-23 福建农林大学 Method for preparing solid fermentation culture medium for bacillus thuringiensis
CN102286413A (en) * 2011-09-03 2011-12-21 福建农林大学 Preparation method of liquid fermentation medium for bacillus thuringiensis
CN102907458A (en) * 2012-10-28 2013-02-06 郑文定 Biological mosquitocide and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
廖先清 等: "1200 ITU/mg Bti浓悬浮剂的研制", 《湖北农业科学》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109593680A (en) * 2018-12-26 2019-04-09 武汉科诺生物科技股份有限公司 A kind of Dipel liquid fermentation medium and its bacterium powder and oil suspending agent
CN109593680B (en) * 2018-12-26 2022-02-22 武汉科诺生物科技股份有限公司 Bacillus thuringiensis liquid fermentation medium and bacterial powder and oil suspension agent thereof

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Application publication date: 20170215