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WO2001091559A2 - Insectifuge, préparation et utilisation - Google Patents

Insectifuge, préparation et utilisation Download PDF

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Publication number
WO2001091559A2
WO2001091559A2 PCT/ZA2001/000059 ZA0100059W WO0191559A2 WO 2001091559 A2 WO2001091559 A2 WO 2001091559A2 ZA 0100059 W ZA0100059 W ZA 0100059W WO 0191559 A2 WO0191559 A2 WO 0191559A2
Authority
WO
WIPO (PCT)
Prior art keywords
insect repellent
culture
medium
insecticidal
formation
Prior art date
Application number
PCT/ZA2001/000059
Other languages
English (en)
Other versions
WO2001091559A3 (fr
Inventor
Pieter Lodewikus Steyn
Original Assignee
Pieter Lodewikus Steyn
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Pieter Lodewikus Steyn filed Critical Pieter Lodewikus Steyn
Priority to AU2001263530A priority Critical patent/AU2001263530A1/en
Publication of WO2001091559A2 publication Critical patent/WO2001091559A2/fr
Publication of WO2001091559A3 publication Critical patent/WO2001091559A3/fr

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Classifications

    • 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
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22—Bacillus
    • A01N63/23—B. thuringiensis

Definitions

  • This invention relates to an insect repellent and the use thereof. This invention further relates to methods of preparing an insect repellent and of repelling insects.
  • Bacillus thuringiensis (B.t.) is a Gram-positive, spore-forming bacterium characterised by parasporal crystalline protein inclusions. These inclusions often appear microscopically as distinctively shaped crystals. The proteins are highly toxic to insects and specific in their toxic activity.
  • an insect repellent comprising a preparation of Bacillus thuringiensis (B.t.) and a carrier medium.
  • the carrier medium may be a liquid.
  • the carrier medium may be solid particles.
  • the carrier medium is a suitable complex medium for sustaining growth of B.t. and for allowing sporulation and formation of the insecticidal parasporal protein crystals of B.t..
  • the medium may include a sufficient amount of proteins or protein derivatives to meet the nitrogen requirements of B.t.
  • the medium may further include a source of energy and/or carbon.
  • the medium may include a buffer for stabilising the pH at such a level as to allow optimal cell production and crystal formation.
  • the insect repellent includes the following: a B.t. culture; peptone; yeast extract;
  • a method of repelling insects including the steps of: providing a B.t. inoculant; - cultivating the B.t. inoculant on a medium which supports sporulation and formation of the insecticidal parasporal protein crystals of the B.t. to obtain a culture; diluting the culture; and applying the diluted culture to a zone from which insects are to be repelled.
  • the step of applying the culture includes the step of spraying the culture on animals, humans, clothes, fruit trees, vegetables, and/or other zones where insects tend to gather.
  • a method of preparing an insect repellent including the steps of: - inoculating a medium which supports sporulation and formation of insecticidal parasporal protein crystals with a B.t. isolate; cultivating the B.t. in the medium to allow sporulation and formation of the insecticidal parasporal protein, to obtain an insecticidal culture; providing a suitable carrier medium; and - mixing the insecticidal culture with the carrier medium.
  • the method may include the further step of diluting the mixture with water.
  • Bacillus thuringiensis (B.t.) is cultivated on a suitable complex medium.
  • the medium is selected to sustain sufficient growth of B.t. and to allow sporulation and formation of the insecticidal parasporal protein crystals of B.t.
  • the medium therefore contains a sufficient amount of proteins or protein derivatives to meet the nitrogen requirements of B.t., as well as a source of energy and carbon.
  • an appropriate buffer e.g. a phosphate buffer, is included in the medium to stabilise the pH at such a level as to allow for optimal cell production and crystal formation.
  • the medium is prepared from the following components:
  • agar-agar After sterilisation, 15 g of agar-agar is mixed with the solution. The mixture is poured into Bijou bottles and allowed to set to provide a slanted solid growth medium.
  • a B.t. culture is maintained on the surface of the solid growth medium.
  • Another litre of the growth medium is prepared, excluding the agar-agar, to provide a sterile liquid growth medium.
  • the liquid medium is inoculated with B.t. by adding a small volume of sterile water (2ml for example) to the B.t. culture on the solid growth medium, and scraping the B.t. culture from said surface with a sterile spatula.
  • the B.t. culture is added to the liquid growth medium, with a sterile Pasteur pipette under aseptic conditions.
  • This culture is grown with aeration at 30 °C for 48 hours, and is then inoculated into 15 to 40 litres of sterile liquid medium in a fermenter under aseptic conditions.
  • This inoculated medium is incubated in an incubator room at 30 °C while being constantly agitated with a mechanical stirring device or sterile air, which also supplies the necessary oxygen for aerobic growth of the bacteria.
  • the required cell and parasporal crystal density (1x10 9 spores per ml) is attained as determined by total spore counts using the Petroff-Hausser bacterial counting chamber and phase contrast microscopy, the culture is distributed into smaller pre-sterilised plastic containers of 250 ml each and sealed under aseptic conditions. This culture is then ready for use as an insect repellant as set out below.
  • the culture in the fermenters is constantly observed by phase contrast microscopy and at the onset of sporulation, an inoculum consisting of aliquots of this culture, e.g. 20 ml quantities, is injected into a suitable presterilised semisolid matrix, which comprises 82g finely comminuted perlite or diatomite and 100ml of the above liquid medium. Prior to inoculation, the semisolid matrix is distributed into high density plastic bags and steam sterilised, sealed and kept until inoculation.
  • Inoculation of the B.t. into the semi-solid matrix is effected by means of volumetric injection of the inoculum with a measuring peristaltic or cylinder automatic pipetting machine. These inoculated sachets are incubated for 7 days at 30 °C. After this period, a viable spore count is made by serial dilution of the contents of a sachet, exposing the series dilution tubes to 65 °C for 10 minutes, and plating 0,1 ml quantities of each dilution on Nutrient Agar in Petri dishes. After incubation of these plates at 30 °C for 48 hours, the number of colonies developing on the agar are counted, taking into account only those plates on which between 30 and 500 colonies developed. Each colony is taken to represent one viable spore. At least 1x10 9 colony forming units per gram are required to constitute an acceptable product.
  • the insect repellent is used as follows:
  • 250 ml of liquid repellent, or 200g of semi-solid repellent, containing spores and crystals, is diluted with 20 to 150 litre water and used for spraying animals, humans, clothes, fruit trees, vegetables, insect breeding places, and/or places where the insects tend to gather.
  • the repellent is mixed with enough water to treat 50 heads of cattle, 40 horses, 200 sheep or goats or 200 ostriches.
  • the animals need not be sprayed such that they become completely wet, but should rather be covered only with a light mist of the repellent.
  • the gathering places of the insects are sprayed with the same preparation as in the case of animals and only the environment should be properly wetted.
  • insects that are repelled by the repellent include gnats or black flies, mosquitoes and the common house fly, but are not limited thereto.
  • insect repellent according to the invention is highly effective in repelling insects.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Pest Control & Pesticides (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

Cette invention a trait à l'emploi du Bacillus thuringiensis comme insectifuge ainsi qu'à un insectifuge renfermant une préparation de ce bacille (B. t.) et un véhicule. L'invention concerne également un procédé de préparation d'un insectifuge, consistant à inoculer un milieu favorable à la sporulation et à la formation de cristaux de protéine parasporale au moyen d'un isolat de B. t., à cultiver ce dernier dans le milieu pour permettre la sporulation et la formation de la protéine parasporale insecticide, à constituer un véhicule approprié et à incorporer la culture insecticide au véhicule. L'invention concerne, de plus, un procédé de lutte contre les insectes consistant à diluer la culture et à répandre cette dilution sur une zone que l'on veut protéger des insectes.
PCT/ZA2001/000059 2000-05-30 2001-05-18 Insectifuge, préparation et utilisation WO2001091559A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001263530A AU2001263530A1 (en) 2000-05-30 2001-05-18 An insect repellent and the preparation and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ZA2000/2692 2000-05-30
ZA200002692 2000-05-30

Publications (2)

Publication Number Publication Date
WO2001091559A2 true WO2001091559A2 (fr) 2001-12-06
WO2001091559A3 WO2001091559A3 (fr) 2002-03-07

Family

ID=25588768

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ZA2001/000059 WO2001091559A2 (fr) 2000-05-30 2001-05-18 Insectifuge, préparation et utilisation

Country Status (2)

Country Link
AU (1) AU2001263530A1 (fr)
WO (1) WO2001091559A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
BE1018722A5 (nl) * 2009-04-17 2011-07-05 Cruysberghs Rudiger Een biologische samenstelling voor het behandelen van huisdieren en hun levensomgeving.
BE1018872A5 (nl) * 2009-08-28 2011-10-04 Cruysberghs Rudiger Afweermiddel tegen muggen.
EP1818405B1 (fr) 2004-04-09 2015-06-03 Monsanto Technology, LLC Compositions et procédés de contrôle d'infestation d'insectes dans des plantes
CN114504000A (zh) * 2022-02-17 2022-05-17 浙江省林业科学研究院 一种环保酵素及其快速制备方法和应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU501727A1 (ru) * 1973-08-31 1976-02-05 Всесоюзный Научно-Исследовательский Институт Микробиологических Средств Защиты Растений И Бактериальных Препаратов Фагоустойчивый штамм 13-э внии бакпрепарат-продуцент энтобактерина
SU1181316A1 (ru) * 1984-05-28 1986-12-30 Всесоюзный научно-исследовательский институт прикладной микробиологии Штамм @ @ @ , @ @ -51-продуцент протеолитических ферментов
DE4442255A1 (de) * 1994-11-28 1996-05-30 Bayer Ag Schädlingsbekämpfungsmittel
JP4132165B2 (ja) * 1997-12-16 2008-08-13 愛知県 繊維の防虫加工方法

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1818405B1 (fr) 2004-04-09 2015-06-03 Monsanto Technology, LLC Compositions et procédés de contrôle d'infestation d'insectes dans des plantes
US10167484B2 (en) 2004-04-09 2019-01-01 Monsanto Technology Llc Compositions and methods for control of insect infestations in plants
US10787680B2 (en) 2004-04-09 2020-09-29 Monsanto Technology Llc Compositions and methods for control of insect infestations in plants
US11492638B2 (en) 2004-04-09 2022-11-08 Monsanto Technology, Llc Compositions and methods for control of insect infestations in plants
US11685930B2 (en) 2004-04-09 2023-06-27 Monsanto Technology, Llc Compositions and methods for control of insect infestations in plants
US12077770B2 (en) 2004-04-09 2024-09-03 Monsanto Technology Llc Compositions and methods for control of insect infestations in plants
BE1018722A5 (nl) * 2009-04-17 2011-07-05 Cruysberghs Rudiger Een biologische samenstelling voor het behandelen van huisdieren en hun levensomgeving.
BE1018872A5 (nl) * 2009-08-28 2011-10-04 Cruysberghs Rudiger Afweermiddel tegen muggen.
WO2011022790A3 (fr) * 2009-08-28 2011-10-27 Rudiger Cruysberghs Anti-moustique
CN102740700A (zh) * 2009-08-28 2012-10-17 鲁迪吉尔·克勒伊斯贝里斯 驱蚊剂
CN114504000A (zh) * 2022-02-17 2022-05-17 浙江省林业科学研究院 一种环保酵素及其快速制备方法和应用
CN114504000B (zh) * 2022-02-17 2023-06-27 浙江省林业科学研究院 一种环保酵素及其快速制备方法和应用

Also Published As

Publication number Publication date
WO2001091559A3 (fr) 2002-03-07
AU2001263530A1 (en) 2001-12-11

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