WO2011026983A1 - Use of beneficial insects in plant protection with pesticides - Google Patents

Use of beneficial insects in plant protection with pesticides Download PDF

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Publication number
WO2011026983A1
WO2011026983A1 PCT/EP2010/063056 EP2010063056W WO2011026983A1 WO 2011026983 A1 WO2011026983 A1 WO 2011026983A1 EP 2010063056 W EP2010063056 W EP 2010063056W WO 2011026983 A1 WO2011026983 A1 WO 2011026983A1
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Prior art keywords
plant protection
chemical plant
fungicides
protection agents
plot
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PCT/EP2010/063056
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French (fr)
Inventor
Veerle Mommaerts
Linde Mieke Erna Besard
Guy Smagghe
Guido Mario Karel Michel Sterk
Christian Ernest Serge Gunther Van Osselaer
Herman Felicien Marie Van Mellaert
Kurt Hans Put
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Biobest Belgium Nv
Vrije Universiteit Brussel
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Application filed by Biobest Belgium Nv, Vrije Universiteit Brussel filed Critical Biobest Belgium Nv
Publication of WO2011026983A1 publication Critical patent/WO2011026983A1/en

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K47/00—Beehives
    • A01K47/06—Other details of beehives, e.g. ventilating devices, entrances to hives, guards, partitions or bee escapes
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01M—CATCHING, TRAPPING OR SCARING OF ANIMALS; APPARATUS FOR THE DESTRUCTION OF NOXIOUS ANIMALS OR NOXIOUS PLANTS
    • A01M1/00—Stationary means for catching or killing insects
    • A01M1/20—Poisoning, narcotising, or burning insects

Definitions

  • the present invention relates to the distribution of pesticides and beneficial organisms in agriculture and horticulture, and is based on the risk assessment of side- effects of chemical plant protection agents, like pesticides herbicides, insecticides and fungicides, on the pollinator B. terrestris . It accordingly provides the use of the pollinator Bombus terrestris, in disseminating chemical plant protection agents in agriculture and horticulture.
  • bacterial biological control agents such as Streptomyces (Smith et al . , Phytopathology, 2009, 99(6) Supl S121) ;
  • Gliocladium or Trichoderma such as Gliocladium or Trichoderma (Smith et al . , Phytopathology, 2009, 99(6) Supl S121; Kovach et al . , Biological Control, 2000, 18, p.235-242; Shafir et al., Eur. J. of Plant Pathology, 2006, 116, p.119-128; Maccagnani et al . , Bulletin of Insectology, 2005, 58(1), p.3-8; and
  • the present invention provides a method for disseminating chemical plant protection agents in agriculture and horticulture, said method comprising the use of pollinators.
  • said method further includes the use of a beehive dispenser, such as the side-by-side passageway dispenser (SSP) or the overlapping passageway dispenser (OP) that are mounted on the exit of the beehive.
  • SSP side-by-side passageway dispenser
  • OP overlapping passageway dispenser
  • the present invention provides the use of pollinators in disseminating chemical plant protection agents in agriculture and horticulture.
  • the chemical plant protection agent can be any known agrochemical typically used for pest and disease control in agriculture and horticulture.
  • the chemical plant protection agents as used herein refer to the various chemical non-natural, i.e. synthetic products used in agriculture and horticulture. These include Fungicides for the control of fungi and oomycetes; Herbicides (e.g. glyphosate) for the control of weeds; Insecticides (e.g.
  • organochlorines organophosphates , carbamates, and pyrethroids
  • organochlorines organophosphates , carbamates, and pyrethroids
  • ovicides substances that kill eggs
  • larvicides substances that kill larvae
  • adulticides substances that kill adults
  • Miticides or acaricides for the control of mites. It in particular relates to insecticides like imidacloprid or Fungicides, such as for example the botryticides ; Frupica (mepanipyrine, 50%, WP) , Anilinopyrimidines (AP) -fungicides, Phenylpyrroles
  • the chemical plant protection agent is a fungicide, such as for example botryticides ; and selected from the group consisting of Frupica (mepanipyrine, 50%, WP) , Anilinopyrimidines (AP) - fungicides, Phenylpyrroles ( PP) -fungicides , Quinone outside inhibitors (Qol ) -fungicides , Methyl benzimidazole carbamates
  • MCC Demethylation inhibitors
  • DMI Demethylation inhibitors
  • Rovral iprodion, 75%WG
  • Signum boscalid+pyraclostrobin, 26%+ 6.7%, WG
  • Sumico diethofencarb+carbendazim
  • BCAs biological control agents
  • disseminated substance as used in the invention is intended to deliver an added utility to the plant (preferably via the flowers) visited by the bees such as reducing certain pest and disease problems.
  • Said substances can be used as such, or in the form of a formulation, including combinations of the aforementioned substances.
  • the substance is in a liquid or powdered formulated, more in particular the formulation is a powdered formulation, e.g. using a powdered carrier.
  • the powdered carrier can be a starch such as corn, starch, talc, dust derived from crushed hulls of nuts, clay dust, or mixtures thereof.
  • the chemical plant protection agent and carrier are present in a ratio ranging from about 100% of plant protection agent to about 50/50 w/w ratio of plant protection agent over carrier.
  • the chemical plant protection agents are present at sublethal concentrations for the pollinators and in particular at their maximum field recommended concentration (MFRC) .
  • the chemical plant protection agents are used ad sublethal concentrations for the pollinators and in particular used at their maximum field recommended concentration (MFRC) .
  • the pollinators used in the aforementioned embodiments are the ones known to the skilled artisan for their pollinating services in agriculture and horticulture and typically include honeybees and bumblebees, more in particular bumblebees .
  • the present study evaluated the risks on bumblebees of six currently used botryticides : Frupica, Rovral, Signum, Sumico, Switch and Teldor, and this was done with use of queenless micro- colonies of 5 workers in the laboratory.
  • the workers were exposed to the different compounds at their respective maximum field recommended concentration (MFRC) and this via three different routes of exposure: dermal contact, and orally via treatment of the drinking sugar water and eating pollen.
  • MFRC maximum field recommended concentration
  • Table 1 Overview of the six chemical botryticides evaluated on their compatibility with B. terrestris. For all botryticides their commercial name, their active ingredient (AI) and fungicide mode of action class, their formulation type and amount of AI, and their MFRC in % formulation and ppm is given.
  • AI active ingredient
  • fungicide mode of action class For all botryticides their commercial name, their active ingredient (AI) and fungicide mode of action class, their formulation type and amount of AI, and their MFRC in % formulation and ppm is given.
  • WG water dispersible granules
  • WP wettable powder
  • Newly born workers were collected from the rearing and placed per five in an artificial nest box made of plastic (15 cm x 15 cm x 10 cm) . In the centre of the nest there was a drinking place and brood area. Under the nest a container with 500 ml sugar water was provided via which bumblebee workers had access by drinking on a cotton wick which contained the sugar water by capillarity. After 1 week, the dominant worker started to produce eggs that develop into males. Per treatment four artificial nests were exposed, and each experiment was two times repeated.
  • the MFRC of each product was prepared in water. Individual bees were topically treated with 50 ⁇ of this aqueous solution on their dorsal thorax with a micropipette . For the oral treatments, bumblebee workers were exposed to 500 ml sugar water (1/1) that was dosed with the respective compound at its MFRC, or to pollen sprayed until saturated with the MFRC of the product in water. In these nests the sugar water and pollen were weekly replaced with freshly prepared material.
  • Table 3 Overview of the sublethal effects on reproduction in workers of Bombus terrestris by the six botryticides tested when treated at their respective MFRC via topical contact or orally via treated sugar water or treated pollen.
  • the data are expressed as mean numbers of drones per nest ⁇ SEM based on 4 artificial nests per treatment and 5 workers per nest each, and the experiment was two times repeated.
  • the method with use of five B. terrestris workers allows to assess lethal and sublethal side-effects on microcolony level and has the advantage to work in a standardized manner under laboratory conditions. Moreover, due to the small size of the microcolonies , consisting of 5 workers per nests, this method is easy in handling and in follow up.
  • the different commercial fungicides tested did not cause lethal or sublethal effects on reproduction following exposure via three different routes. This showed that both the larval stage, which developed later into drones and the adult stage (workers) were not affected by the fungicide because pollen is mainly consumed by larvae whereas sugar water is the energy source of bumblebee workers (Heinrich, 1979) .
  • cinerea management is still relaying on chemical control mainly, although the number of reports on resistance development in fungal pathogens against fungicides is increasing (Dianez et al . , 2002; Leroux et al . , 2002; Bardas et al . , 2008; Kretschmer et al . , 2009) . Moreover, the use of fungicides was also shown to adversely affect pollen germination which in turn results in reduced fruit formation (Kovach et al . , 2000) . As alternatives for chemical control, various biocontrol agents have been used in the past against B. cinerea by a spray application and by vectoring.
  • the aim of the present study was to evaluate the efficacy of Bombus terrestris Linnaeus (Hymenoptera : Apidae) to disseminate pesticides, and in particular Signum ® by use of a hive dispenser into strawberry flowers in the greenhouse.
  • Bombus terrestris Linnaeus Heymenoptera : Apidae
  • Signum ® a hive dispenser into strawberry flowers in the greenhouse.
  • the efficiencies of transport/dissemination into the flowers and the subsequent control of B. cinerea were evaluated by scoring the healthy red strawberry fruits before/at harvest (pre-harvest ) and also after incubation during 2 days at conditions for optimal Botrytis growth (post-harvest) .
  • B. terrestris can be used effectively to disseminate the botryticide Signum ® (WG) in a greenhouse, and provides an efficient control of B. cinerea in strawberry plants.
  • WG botryticide Signum ®
  • all flowers developed into mature fruits while the traditional inoculation method a large percentage of undeveloped fruit was found on average 58 ⁇ 3% (see results below) .
  • This further advantage clearly supports the use of bumblebees for the dissemination of botryticiden in horticulture and agriculture, as a cheaper and more efficient alternative.
  • This semi-field test was conducted in a greenhouse (15m x 8m) located at Biobest. This greenhouse was divided into three compartments ( Figure 1) .
  • the first compartment (bottom left next to the entrance compartment) or Botrytis (B) Plot (2x3m) includes 60 strawberry plants .
  • the second compartment (top left) or Botrytis Signum® (BS) plot (2x3m) includes 60 strawberry plants.
  • the third compartment (large rectangle right) or Botrytis-Signum®-Hommel (BSH) plot (13x8m) contains 360 strawberry plants.
  • the compartments are each separated by fine meshed tents to avoid contamination between the different plots.
  • Per plot the strawberry plants were arranged in double rows, each with 6 plants per tray. The plants were watered twice weekly .
  • the plants in this plot were manually inoculated with a spores solution of B. cinerea.
  • the manual infection of the flowers with B. cinerea was repeated every 3 days. This is a real frequency, since the life of a strawberry flower is 3 days .
  • the plants in the BS-plot were also manually inoculated with a spores solution of B. cinerea. Additionally and different from the B-plot, in the BS-plot the flowers of the strawberry plants were each morning manually inoculated with 20 ⁇ 1 Signum solution (MFRC) .
  • MFRC Signum solution
  • This plot uses a standard bumblebee nest (1 queen and 50 workers) with the miniature dispenser (20x5cm) containing a mixture of Signum® with an appropriate carrier substance at a 50/50 w/w ratio.
  • the miniature dispenser (20x5cm) containing a mixture of Signum® with an appropriate carrier substance at a 50/50 w/w ratio.
  • two different carriers were tested, i.e. potato starch and corn starch.
  • the total product mix was weighed at the beginning and the end of the test.
  • the charts in Figure 2 show the average number of CFU of B. cinerea per flower again for the three plots for Signum® (WG) mixed with the excipients potato starch (Fig. 2A) and corn starch (Fig. 2B) .
  • CFU average number of B. cinerea spores
  • a significant reduction in the average number of B. cinerea spores (CFU) could be observed, i.e. 52 ⁇ 1% reduction in the Signum®/potato starch mixture and 83 ⁇ 2% reduction in the Signum®/corn starch mixture.
  • This reduction, both for potato starch (p ⁇ 0.05) and for corn starch (p ⁇ 0.05) differ significantly from the control plot (B-plot) , where 100 ⁇ 0% infection was found.

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Pest Control & Pesticides (AREA)
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  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
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Abstract

The present invention relates to the distribution of pesticides and beneficial organisms in agriculture and horticulture, and is based on the risk assessment of side-effects of chemical plant protection agents, like pesticides herbicides, insecticides and fungicides, on the pollinator Bombus Terrestris. It accordingly provides the use of the pollinator Bombus Terrestris, in disseminating chemical plant protection agents in agriculture and horticulture.

Description

SE OF BENEFICIAL INSECTS IN PLANT PROTECTION WITH PESTICIDES
Field of the Invention
The present invention relates to the distribution of pesticides and beneficial organisms in agriculture and horticulture, and is based on the risk assessment of side- effects of chemical plant protection agents, like pesticides herbicides, insecticides and fungicides, on the pollinator B. terrestris . It accordingly provides the use of the pollinator Bombus terrestris, in disseminating chemical plant protection agents in agriculture and horticulture.
Background to the Invention
Flowers are often important pathways of plant disease infection. The pathogen infects them symptomless under favorable conditions, and progressively colonizes other tissues or fruits. Disease symptoms become visible when infected tissues ripen, senesce or die, as for example in the case of Botrytis cinerea. Effective biological plant disease control depends both on the use of suitable antagonistic strains, and on the methods and strategies for introducing, promoting and maintaining the antagonist in the crop (Sutton and Peng, Annual Review of Phytopathology, 1993, 31, p. 473-493) .
Although chemical substances are very suitable for eliminating pathogenic infections, the use of chemical plant protection agents comes into conflict with the general application of pollinators like bumblebees and honeybees in modern agriculture. During and for a certain period after the use of chemical plant protection agents in the green house, pollinators should be kept aside. To address the aforementioned conflict of chemical plant protection agent with the use of pollinators in modern agriculture, a lot of effort has been put forward in the development of biological control agents (BCAs) . This method of controlling pests (including insects, mites, weeds and plant diseases) , is based on the reduction of pest populations by natural enemies, such as for example based on predation, parasitism, herbivory, or other natural mechanisms and typically involves an active human role. Pest control through biological control agents is not only based on the conservation of the natural enemies in the field, but also includes an active and supplementary release of said natural enemies within the field.
In said aspect it has previously been shown that pollinators like honeybees and bumblebees can efficiently be used in the dissemination of biological control agents like;
- microbial, i.e. bacterial biological control agents such as Streptomyces (Smith et al . , Phytopathology, 2009, 99(6) Supl S121) ;
- fungal antagonists such as Gliocladium or Trichoderma (Smith et al . , Phytopathology, 2009, 99(6) Supl S121; Kovach et al . , Biological Control, 2000, 18, p.235-242; Shafir et al., Eur. J. of Plant Pathology, 2006, 116, p.119-128; Maccagnani et al . , Bulletin of Insectology, 2005, 58(1), p.3-8; and
- entomopathogenic fungi such as Beauveria (Al-mazra' awi et al., Environ. Entomol., 2006, 35(6), p.1569-1577; Al- mazra' awi et al . , Biological Control, 2007, 36, p.89-97; Kapongo et al . , Biological Control, 2008, p.508-514; Kapongo et al., Biocontrol, 2008, 53, p.797-812).
Notwithstanding the efficient application of pollinators in the dissemination of biological control agents, and in view of the generally accepted paradigm that traditional chemical plant protection agents are toxic and incompatible with the general application of pollinators like bumblebees and honeybees in modern agriculture, up till today no effort has been done in trying to use pollinators in the dissemination of the traditional chemical plant protection agents. One of the elements that strengthens the aforementioned paradigm is the observation that residues of chemical plant protection agents on the plants and in the greenhouse, negatively affect the foraging behavior of bees (Desneux et al . , Annual Review of Entomology, 2007, 52, p.81-106) .
For practical reasons, it would be beneficial if the dissemination of the chemical plant protection agents and the use of pollinators could be harmonized.
It is accordingly a general object of the invention to provide a novel method in distributing chemical plant protection agents in agriculture and horticulture.
Description of the Invention
In a first aspect the present invention provides a method for disseminating chemical plant protection agents in agriculture and horticulture, said method comprising the use of pollinators. In a further aspect, said method further includes the use of a beehive dispenser, such as the side-by-side passageway dispenser (SSP) or the overlapping passageway dispenser (OP) that are mounted on the exit of the beehive.
In a second aspect, the present invention provides the use of pollinators in disseminating chemical plant protection agents in agriculture and horticulture.
In the aforementioned aspects of the invention, the chemical plant protection agent can be any known agrochemical typically used for pest and disease control in agriculture and horticulture. As is evident from the foregoing and the examples hereinafter, the chemical plant protection agents as used herein refer to the various chemical non-natural, i.e. synthetic products used in agriculture and horticulture. These include Fungicides for the control of fungi and oomycetes; Herbicides (e.g. glyphosate) for the control of weeds; Insecticides (e.g. organochlorines , organophosphates , carbamates, and pyrethroids) for the control of insects - these can be ovicides (substances that kill eggs) , larvicides (substances that kill larvae) or adulticides (substances that kill adults) ; and Miticides or acaricides for the control of mites. It in particular relates to insecticides like imidacloprid or Fungicides, such as for example the botryticides ; Frupica (mepanipyrine, 50%, WP) , Anilinopyrimidines (AP) -fungicides, Phenylpyrroles
( PP) -fungicides , Quinone outside inhibitors (Qol)- fungicides, Methyl benzimidazole carbamates (MBC) - fungicides, Demethylation inhibitors (DMI ) -fungicide, Rovral
(iprodion, 75%WG) , Signum (boscalid+pyraclostrobin, 26%+ 6.7%, WG) , Sumico (diethofencarb+carbendazim,
25.5%+25.5%, WP) , Switch (cyprodinil+fludioxonil, 37.5%+25%, WG) and Teldor (fenhexamid, 50%, WG) ; in particular the chemical plant protection agent is a fungicide, such as for example botryticides ; and selected from the group consisting of Frupica (mepanipyrine, 50%, WP) , Anilinopyrimidines (AP) - fungicides, Phenylpyrroles ( PP) -fungicides , Quinone outside inhibitors (Qol ) -fungicides , Methyl benzimidazole carbamates
(MBC) -fungicides , Demethylation inhibitors (DMI ) -fungicide, Rovral (iprodion, 75%WG) , Signum (boscalid+pyraclostrobin, 26%+ 6.7%, WG) , Sumico (diethofencarb+carbendazim,
25.5%+25.5%, WP) , Switch (cyprodinil+fludioxonil, 37.5%+25%, WG) and Teldor (fenhexamid, 50%, WG) ; more in particular Rovral (iprodion, 75%WG) , or Signum
(boscalid+pyraclostrobin) . It explicitly excludes biological control agents (BCAs) like bacterial biological control agents, fungal antagonists, and entomopathogenic fungis, i.e. agents based on the natural (non-synthetic) enemies of the pest to be controlled.
While it is well known in the art that bee hives without dispenser and disseminated substance included therein are useful for pollination of certain plants, the disseminated substance as used in the invention is intended to deliver an added utility to the plant (preferably via the flowers) visited by the bees such as reducing certain pest and disease problems.
Said substances can be used as such, or in the form of a formulation, including combinations of the aforementioned substances. In a particular embodiment, the substance is in a liquid or powdered formulated, more in particular the formulation is a powdered formulation, e.g. using a powdered carrier. The powdered carrier can be a starch such as corn, starch, talc, dust derived from crushed hulls of nuts, clay dust, or mixtures thereof. In such formulation, the chemical plant protection agent and carrier are present in a ratio ranging from about 100% of plant protection agent to about 50/50 w/w ratio of plant protection agent over carrier. In any instance, and independent of the formulation used, the chemical plant protection agents are present at sublethal concentrations for the pollinators and in particular at their maximum field recommended concentration (MFRC) .
Consequently, in the methods of the present invention, the chemical plant protection agents are used ad sublethal concentrations for the pollinators and in particular used at their maximum field recommended concentration (MFRC) .
The pollinators used in the aforementioned embodiments are the ones known to the skilled artisan for their pollinating services in agriculture and horticulture and typically include honeybees and bumblebees, more in particular bumblebees .
This invention will be better understood by reference to the Experimental Details that follow, but those skilled in the art will readily appreciate that these are only illustrative of the invention as described more fully in the claims that follow thereafter. Additionally, throughout this application, various publications are cited. The disclosure of these publications is hereby incorporated by reference into this application to describe more fully the state of the art to which this invention pertains. EXAMPLES
EXAMPLE 1 . RISK ASSESSMENT OF SIDE-EFFECTS OF BOTRYTICIDES ON THE POLLINATOR BOMBUS TERRESTRIS
To date bumblebees, Bombus terrestris (L.), are more and more used in agriculture and horticulture for their pollinating services. But in addition to a good pollination, cultivators also need to control important plant pathogens as grey mold Botrytis cinerea (L.) . Although fungicides are primarily directed to the control of fungi and oomycetes, these chemistries represent different biological activities, and it is not self-evident to combine a chemical treatment with a pollinating services. Therefore, the present study evaluated the risks on bumblebees of six currently used botryticides : Frupica, Rovral, Signum, Sumico, Switch and Teldor, and this was done with use of queenless micro- colonies of 5 workers in the laboratory. The workers were exposed to the different compounds at their respective maximum field recommended concentration (MFRC) and this via three different routes of exposure: dermal contact, and orally via treatment of the drinking sugar water and eating pollen. We scored potential lethal effects on worker survival, and sublethal side-effects on next brood with larval survival and drone formation over a period of 8 weeks .
Under the tested conditions, there were no lethal and sublethal effects, supporting that the tested botryticides are safe to be used in combination with B. terrestris. However, contact treatment with Frupica showed a higher nest reproduction, and potential mechanisms are discussed. 1.1. Materials and Methods
1.1.1. Insects
All experiments were performed with workers of the bumblebee B. terrestris obtained from a continuous mass rearing (Biobest NV, Westerlo, Belgium) and done under standardized laboratory conditions of 28-30°C, 60-65% RH and darkness. The insects were provided ad libitum with commercial sugar water and pollen (Apihurdes, Extremura, Spain) as energy and protein source, respectively.
1.1.2. Products
In this study six botryticides were tested (Table 1) . Each product was stored in accordance with the manufacturers' guidelines .
Table 1. Overview of the six chemical botryticides evaluated on their compatibility with B. terrestris. For all botryticides their commercial name, their active ingredient (AI) and fungicide mode of action class, their formulation type and amount of AI, and their MFRC in % formulation and ppm is given.
Commercial AI (fungicide mode of action Formulation type3; MFRC (%) MFRC (mg name class) mg Al/g AI/1)
Frupica mepanipyrine (AP-fungicides) WP; 50 0.06 300 ovral iprodion (dicarboxamides) WG; 75 0.2 1500
Signum boscalid + pyraclostrobin WG; 26 0.2 520
(carboximides + Qol-fungicides) WG; 6.7 134
Sumico carbendazin + diethofencarb WP; 25.5 0.2 510
(N-phenyl carbamates + MBC- WP; 25.5 510 Commercial AI (fungicide mode of action Formulation type3; MFRC (%) MFRC (mg name class) mg Al/g AI/1)
fungicides)
Switch cyprodinil + fludioxonil WG; 37.5 0.1 375
(AP-fungicides + PP-fungicides) WG; 25 250
Teldor fenhexamid (hydroxyanilides) WG; 50 0.15 750
WG = water dispersible granules; WP = wettable powder
1.1.3. Insect bioassay to assess side-effects with bumblebee workers
Newly born workers were collected from the rearing and placed per five in an artificial nest box made of plastic (15 cm x 15 cm x 10 cm) . In the centre of the nest there was a drinking place and brood area. Under the nest a container with 500 ml sugar water was provided via which bumblebee workers had access by drinking on a cotton wick which contained the sugar water by capillarity. After 1 week, the dominant worker started to produce eggs that develop into males. Per treatment four artificial nests were exposed, and each experiment was two times repeated.
Adults workers were exposed to the product at their respective MFRC via three different routes: via contact by topical application, and orally via treated sugar water and via eating treated pollen.
For the contact treatment the MFRC of each product was prepared in water. Individual bees were topically treated with 50 μΐ of this aqueous solution on their dorsal thorax with a micropipette . For the oral treatments, bumblebee workers were exposed to 500 ml sugar water (1/1) that was dosed with the respective compound at its MFRC, or to pollen sprayed until saturated with the MFRC of the product in water. In these nests the sugar water and pollen were weekly replaced with freshly prepared material.
For the water controls, bumblebees were treated topically with water alone or fed on untreated sugar water or on pollen treated with water only; in these treatments 0-15% worker mortality was recorded. In addition, as positive controls, workers were dosed with the neonicotinoid insecticide formulation imidacloprid (Confidor; Bayer CropSciences , Monheim, Germany) at its MFRC (200 ppm) via the three exposure methods; in all cases this resulted in 100% worker mortality.
In the different artificial nests, worker survival (acute toxicity) was evaluated daily for the 3 days post-treatment, and then on a weekly basis for a period of 11 weeks (chronic toxicity) . The treatments were scored in accordance with IOBC classification for extended laboratory testing:
IOBC classification for extended laboratory testing
Mortality % IOBC class
< 25 1 = non-toxic
25 - 50 2 = weakly toxic
50 - 70 3 = moderately toxic
> 75 4 = highly toxic
Subsequently the adverse effects on the reproduction were monitored on a weekly basis for 8 weeks or 11 weeks by scoring the number of drones produced per nest. 1.1.4. Statistical analysis
All data were analyzed for normal distribution by Kolmogorov-Smirnov (p = 0.05). Unless otherwise stated, normal distribution was confirmed and then data were analyzed by one-way analysis of variance (ANOVA) . Means ± SEM were separated using a post-hoc Tukey-Kramer test (a = 0.05) in SPSS 16.0 (SPSS Inc., Chicago, IL) .
1.2. Results
1.2.1. Acute toxicity on the bumblebee workers
None of the six botryticides , when tested at their respective MFRC, exhibited any lethal effect against the bumblebee workers during the first 72 h after treatment. The response was regardless of the different routes of exposure: by contact or orally via treated sugar water or contaminated pollen. In all cases, 0 ± 0% mortality was scored as in the negative controls (data not shown) .
1.2.2. Chronic toxicity on the bumblebee workers
After 11 weeks for all botryticides, the mortality rates did not exceed 25% at the end of the test period and, as a consequence, these botryticides were scored as class 1 = not toxic (IOBC) . In the controls the worker mortality scored after 11 weeks ranged between 5 - 10% (topical contact), 0 - 5% (sugar water) and 0 - 10% (pollen) . Table 2. Overview of the acute and chronic toxicity of the six botryticides tested on B. terrestris after 11 weeks when exposure to their respective MFRC by topical contact or orally via treated sugar water or treated pollen.
Mean chronic worker mortality ± SEM (IOBC class)*
BOTRYTICIDES Contact Sugar water Pollen
Frupica 0.0 ± 0.0 (1) 2.8 ± 2.0 (1) 5.8 ± 3.4 (1) ovral 0.1 ± 0.1 (1) 10.6 ± 7.5 (1) 3.8 ± 2.7 (1)
Signum 2.6 ± 2.6 (1) 0.2 ± 0.1 (1) 3.0 ± 2.1 (1)
Sumico 0.1 ± 0.1 (1) 0.2 ± 0.1 (1) 0.0 ± 0.0 (1)
Switch 1 1.0 ± 0.4 (1) 0.0 ± 0.0 (1) 0.0 ± 0.0 (1)
Teldor 16.8 ± 16.8 (1) 5.2 ± 0.1 (1) 0.0 ± 0.0 (1)
* Data are given as mean percentage corrected mortality ± SEM after use of Schneider-Orelli formula. The worker mortality in the control groups was 5% (topical contact), 10% (sugar water) and 5% (pollen) after 11 weeks. The mean worker mortality is followed between brackets by the IOBC classification: 1 = not toxic; 2 = weekly toxic; 3 = moderately toxic; 4 = highly toxic.
1.2.3. Sublethal effects on reproduction
For the three different routes of exposure, treatment with MFRC of the six tested botryticides did not cause detrimental effects on reproduction as the mean number of drones was not significantly different (p > 0.05) after 8 weeks as compared with the controls (Table 3) .
Table 3. Overview of the sublethal effects on reproduction in workers of Bombus terrestris by the six botryticides tested when treated at their respective MFRC via topical contact or orally via treated sugar water or treated pollen.
Mean number of drones ± SEM
BOTRYTICIDES * Contact Sugar water Pollen
Frupica 35.8 ± 3.2 b 27.1 ±1.1 a 23.3 ± 0.3 a
Rovral 22.0 ± 2.3 a 24.4 ±1.1 a 21.8 ± 0.2 a
Signum 26.7 ± 1.0 ab 24.8 ±3.5 a 23.1 ± 1.1 a
Sumico 28.0 ± 1.0 ab 30.1 ± 1.4 a 25.4 ± 2.9 a
Switch 23.4 ± 5.6 a 32.3 ± 2.5 a 25.1 ± 0.6 a Mean number of drones ± SEM
BOT YTICIDES * Contact Sugar water Pollen
Teldor 29.4 ± 2.6 ab 27.0 ± 0.5 a 22.8 ± 1.3 a
Control 26.0 ± 0.8 ab 27.5 ± 1.2 a 26.0 ± 0.0 a
The data are expressed as mean numbers of drones per nest ± SEM based on 4 artificial nests per treatment and 5 workers per nest each, and the experiment was two times repeated.
* ANOVA resulted in 2 groups for contact exposure (F = 4.574; df = 48; p = 0.001), in 1 group for sugar water exposure (F = 2.174; df = 47; p = 0.063), and in 1 group for pollen exposure (F = 0.951 ; df = 52; p = 0.470). Values per route of exposure that are followed by a different letter (a - b) are significantly different {post-hoc Tukey-Kramer test with a = 0.05).
1.3. Discussion
The method with use of five B. terrestris workers allows to assess lethal and sublethal side-effects on microcolony level and has the advantage to work in a standardized manner under laboratory conditions. Moreover, due to the small size of the microcolonies , consisting of 5 workers per nests, this method is easy in handling and in follow up.
The different commercial fungicides tested did not cause lethal or sublethal effects on reproduction following exposure via three different routes. This showed that both the larval stage, which developed later into drones and the adult stage (workers) were not affected by the fungicide because pollen is mainly consumed by larvae whereas sugar water is the energy source of bumblebee workers (Heinrich, 1979) .
However it should be remarked that contact exposure to the MFRC (300 ppm) of Frupica (AP-fungicide) tended to stimulate the reproduction, although the effect was not significant (p > 0.05) when compared with the control. Similar, Cabrera et al . (2004) reported a positive effect on the total amount of eggs produced and on the oviposition rate of female mites following contact exposure to the AI fosetyl-Al (phosphonate fungicide) . In the past, mechanistic studies have shown that oral administration of mepanipyrim (containing Frupica) at a dose of 4000 ppm blocked the intracellular transport of lipoproteins from the Golgi to the cell surface in rat hepatocytes (Terada et al . , 1999) and that mepanipyrim acted as an inhibitor of Golgi dispersion (Nakamura et al . , 2003) . However, how this action provoked the observed positive effect on the reproduction is not known.
The current effects agree with the few studies conducted on side-effects of fungicides on Bombus . For example, the fungicide captan was reported safe for B. terrestris as no effects were found after exposure on pollen consumption, oviposition, larval death and drone production (Malone et al . , 2007) . This is also consistent with the findings of the study of Gradisch et al . (2010) who exposed Switch (AP- and PP-fungicide) , Milstop (not classified according to FRAC) and Nova (DMI-fungicide) to B. impatiens . Here, no worker mortality was reported upon direct contact to Switch and Nova with the highest dose tested (1.0 g/1) after 72 h. Also, the same authors found no sublethal effects at a concentration of 521 mg/1 Switch, 136 mg/1 Nova and 476 mg/1 Milstop using microcolonies consisting of 3 workers and commercial hives comprising a queen, her brood and 50 workers (Gradish et al . , 2010). These concentrations resemble the recommended rate for greenhouse use or in case a range of concentrations is given on the product label the middle rate was tested.
Based on the data obtained in this study the tested botryticides at the MFRC are probably safe, but before a final conclusion can be drawn on compatibility more tests are needed. Here it is necessary to assess also potential sublethal risks on the foraging activity. Moreover, due to a lack of information on the future of residues, are they stored in the brood or metabolized, care is needed when applying fungicides. Indeed, Skerl et al . (2009) detected a residue of 0.01 mg/kg difenoconazole (DMI-fungicide) in pollen loads of Apis mellifera Linnaeus (Hymenoptera : Apidae) at 1 day after spray treatment in the orchard. However, at longer time points, i.e. 6-10 days after treatment, the residues were below the detection limit of 0.01 mg/kg (Skerl et al . , 2009).
In addition, in the pollinator-and-vector technology flowers are a reservoir of product residues and fungicides. Consequently, it is likely that returning pollinators deposit a small amount of the fungicides in their nest. Therefore, risk assessments of pesticides on bees/bumblebees need to take this route of exposure into account before drawing any conclusions about their suitability for use in IPM programs .
EXAMPLE 2. SEMI-FILED TEST - I OCULATION OF STRAWBERRY PLANTS WITH BOTRYTICIDES TO SUPPRESS THE PLANT PATHOGEN BOTRYTIS CINEREA USING THE POLLINATOR BOMBUS TERRESTRIS.
Strawberry (Fragaria x ananassa (Weston) Duchesne ex Rozier (Rosales: Rosaceae) ) is a fruit crop grown worldwide but diseases like Botrytis cinerea Pers . : Fr . (Helotiales: Sclerotiniaceae) are limiting its yield. Damage by this pathogen occurs at flowering when conidia originating from infested crop debris or dispersed by wind start to infect the petals, stamens, pistils and then colonize the fruits. In the field, disease symptoms become visible when berries are ripening and thus disease management strategies are needed at flowering. To date, B. cinerea management is still relaying on chemical control mainly, although the number of reports on resistance development in fungal pathogens against fungicides is increasing (Dianez et al . , 2002; Leroux et al . , 2002; Bardas et al . , 2008; Kretschmer et al . , 2009) . Moreover, the use of fungicides was also shown to adversely affect pollen germination which in turn results in reduced fruit formation (Kovach et al . , 2000) . As alternatives for chemical control, various biocontrol agents have been used in the past against B. cinerea by a spray application and by vectoring.
In this context, the aim of the present study was to evaluate the efficacy of Bombus terrestris Linnaeus (Hymenoptera : Apidae) to disseminate pesticides, and in particular Signum® by use of a hive dispenser into strawberry flowers in the greenhouse. Here, the efficiencies of transport/dissemination into the flowers and the subsequent control of B. cinerea were evaluated by scoring the healthy red strawberry fruits before/at harvest (pre-harvest ) and also after incubation during 2 days at conditions for optimal Botrytis growth (post-harvest) .
It was found that B. terrestris can be used effectively to disseminate the botryticide Signum ® (WG) in a greenhouse, and provides an efficient control of B. cinerea in strawberry plants. Moreover, when compared to the traditional 'manual' dissemination of the botryticide, in the inoculation using B. terrestris, all flowers developed into mature fruits, while the traditional inoculation method a large percentage of undeveloped fruit was found on average 58 ± 3% (see results below) . This further advantage clearly supports the use of bumblebees for the dissemination of botryticiden in horticulture and agriculture, as a cheaper and more efficient alternative.
2.1. Materials and Methods
In this experiment, it was tested in a semi-field test whether boscalid (26.7%) + pyraclostrobin (6.7%) (= Signum®) could be disseminated by B. terrestris, through the dispensing system in a strawberry crop of the genus Elsanta in order to effectively combat the gray mold B. cinerea .
As a test plant is chosen to work with strawberry plants of the genus Elsanta. The conservatory was kept at a constant temperature of 20 °C and data loggers for relative humidity and light intensity were present in the greenhouse. 2.1.1. Insects
All experiments were performed with workers of the bumblebee Bombus terrestris obtained from a continuous mass rearing (Biobest NV, Westerlo, Belgium) and done under standardized laboratory conditions of 28-30°C, 60-65% RH and darkness. The insects were provided ad libitum with commercial sugar water and pollen (Apihurdes, Extremura, Spain) as energy and protein source, respectively.
2.1.2. Experimental set-up of the greenhouse
This semi-field test was conducted in a greenhouse (15m x 8m) located at Biobest. This greenhouse was divided into three compartments (Figure 1) . The first compartment (bottom left next to the entrance compartment) or Botrytis (B) Plot (2x3m) includes 60 strawberry plants . The second compartment (top left) or Botrytis Signum® (BS) plot (2x3m) includes 60 strawberry plants. The third compartment (large rectangle right) or Botrytis-Signum®-Hommel (BSH) plot (13x8m) contains 360 strawberry plants.
The compartments are each separated by fine meshed tents to avoid contamination between the different plots. Per plot, the strawberry plants were arranged in double rows, each with 6 plants per tray. The plants were watered twice weekly .
In the B-plot and the BS-plot 30 non-pollinated flowers were scattered within the compartment and labeled with strips (depending on the treatment different colored strips were used) . In the BHS-plot 50 plants were randomly labeled with strips of different colors (one color per round) .
After 3 days in the B-plot and BS-plot 10 flowers; and in the BHS-plot 15 innoculated flowers were collected randomly throughout the plot. The flowers were individually washed to determine the numbers of Colony Forming Units (CFU) of B. cinerea present per flower. Each sample was gently shaken on a rotary shaker in 15 ml of physiological solution for 60 min. Then, a 10-fold serial dilution was made of the aqueous solution and 100 μΐ was put on Potato Dextrose Agar (PDA) . This was repeated twice per sample. For B. cinerea, the numbers of CFU were scored after 3 days of incubation at 22°C.
The remaining flowers were left to develop into fruits, and subsequently scored for infection with B. cinerea through a retention test during which the fruits were kept in a refrigerator at 4°C. The 3 day cycle was repeated two times (i.e. there were 3 rounds of flower harvesting in total) .
2.1.3. Treatment in the B-plot
The plants in this plot, were manually inoculated with a spores solution of B. cinerea. The manual infection of the flowers with B. cinerea was repeated every 3 days. This is a real frequency, since the life of a strawberry flower is 3 days .
2.1.4. Treatment in the BS-plot
In analogy with the plants in the B-plot, the plants in the BS-plot were also manually inoculated with a spores solution of B. cinerea. Additionally and different from the B-plot, in the BS-plot the flowers of the strawberry plants were each morning manually inoculated with 20μ1 Signum solution (MFRC) .
2.1.5. Treatment in the BSH-plot
This plot uses a standard bumblebee nest (1 queen and 50 workers) with the miniature dispenser (20x5cm) containing a mixture of Signum® with an appropriate carrier substance at a 50/50 w/w ratio. In the present experiment two different carriers were tested, i.e. potato starch and corn starch.
To determine the amount of product carried around during the entire duration of the test, the total product mix was weighed at the beginning and the end of the test.
In the BSH plot, the Signum®/carrier mixture was disseminated by the bumblebees during daytime. Inoculation with B . cinerea trace solution occurred at night in a similar manner as described under 3.1.3.
2.2. Results
2.2.1. Direct determination of the CFU B. cinerea / flower
For Signum® (WG) mixed with potato starch the average number of inoculated B. cinerea spores in each of the three plots was 4,6.105 ± 2,7.105; for Signum® mixed with corn starch the average number of inoculated B. cinerea spores in each of the three plots was 2,2.105 ± 0,7.10s. Both inoculation average quantities are therefore in the same order of magnitude and are not significantly different.
The charts in Figure 2 show the average number of CFU of B. cinerea per flower again for the three plots for Signum® (WG) mixed with the excipients potato starch (Fig. 2A) and corn starch (Fig. 2B) . For both carriers in the BSH plot a significant reduction in the average number of B. cinerea spores (CFU) could be observed, i.e. 52 ± 1% reduction in the Signum®/potato starch mixture and 83 ± 2% reduction in the Signum®/corn starch mixture. This reduction, both for potato starch (p <0.05) and for corn starch (p <0.05) differ significantly from the control plot (B-plot) , where 100 ± 0% infection was found.
2.2.2. Infection rate of the fruit after the storage test
Of the 30 labeled flowers in the B-plot and BS-plot; and of the 50 labeled flowers in the BSH-plot, respectively 20 flowers (B-plot and BS-plot) and 35 flowers (BSH-plot) were left to develop into fruits, and subsequently scored for infection with B. cinerea through a retention test during which the fruits were kept in a refrigerator at 4 °C.
In both instances, there is a clear increase in the number of non-infected fruits for the treatment with the two Signum®/carrier mixtures when disseminated by the bumblebees. The results on the mean numbers of non-infected fruits for the different treatments and in the different plots are summarized in table 4 below. Table 4. Overview of the mean number of non-infected fruits after the different treatment methods with the botryticide Signum® with two different carrier substances .
Mean percentage of non-infected fruits ± SEM
BOTRYTICIDE B-plot BS-plot BSH-plot
A Signum®/potato starch 55 ± 1 b 85.0 ± 0 a 66 ± 0 b
B Signum®/corn starch 55 ± 1 a 74 ± 1 a 60 ± 0 a
The data are expressed as mean percentage of non-infected fruits ± SEM. A. Analysis of results using ANOVA resulted in two groups (F = 56.652, df = 5, p = 0.004). Mean values followed by different letters (a-b) are significantly different after Tukey post hoc p = 0.05; ± SEM. B. Analysis of results using ANOVA resulted in one group (F = 2.159, df = 5, p = 0.263). Mean values followed by same letter (a) are significantly equal after post-hoc Tukey p = 0.05; ± SEM.
2.2.2. Number of fully developed mature fruits
In the aforementioned paragraph, all flowers treated during the test were taken into account, even in case said flowers didn't developed in fully developed mature fruits.
Looking at the number of fully developed mature fruits all of the scored flowers in the BSH-plot (independent of the Signum®/carrier mixture used) did develop into fully developed mature fruits (100 ± 0%) . In the B-plots the average number of fully developed mature fruits equaled 71 ± 9% and for the BS-plots only a small number of flowers (42 ± 7%) fully developed in mature fruits.
References
Heinrich B (1979) Bumblebee Economics. Harvard University Press, London, England, pp 245.
Ana R. Cabrera, Raymond A. Cloyd, Edmond R. Zaborski (2004) Effects of Greenhouse Pesticides on the Soil-Dwelling Predatory Mite Stratiolaelaps scimitus (Acari:
Mesostigmata: Laelapidae) Under Laboratory Conditions. Journal of Economic
Entomology 97(3):793-799.
Terada M, Mizuhashi F, Murata K, Tomita T (1999) Mepanipyrim, a new fungicide, inhibits intracellular transport of very low density lipoprotein in rat hepatocytes. Toxicol Appl Pharmacol 154: 1 -11.
Nakamura M, Kono Y, Takatsuki A (2003) Mepanipyrim, a novel inhibitor of pharmacologically induced Golgi dispersion. Biosci Biotechnol Biochem 67: 139-150.
Malone LA, Scott-Dupree CD, Todd JH, Ramankutty P (2007) No sublethal toxicity to bumblebees, Bombus terrestris, exposed to Bt-corn pollen, captan and novaluron. NZJ Crop Hortic Sci 35:435-439.
Gradish AE, Scott-Dupree DC, Shipp L, Harris CR, Ferguson G (2010) Effect of reduced risk pesticides for use in greenhouse vegetable on Bombus impatiens (Hymenoptera: Apidae). Pest Manag Sci 66: 142-146.
Skerl MIS, Bolta SV, Cesnik HB, Gregorc A (2009) Residues of pesticides in honeybee (Apis mellifera carnica) bee bread and in pollen loads from treated apple orchards. Bull Environ Contam Toxicol 83: 374-377.
Dianez F, Santos M, Blanco R, Tello JC (2002) Fungicide resistance in Botrytis cinerea isolates from strawberry crops in Huelva (southwestern Spain). Phytoparasitica 30: 529-534.
Leroux P, Fritz R, Debieu D, Albertini C, Lanen C,Bach J, Gredt M, Chapeland F (2002) Mechanisms of resistance to fungicides in field strains of Botrytis cinerea. Pest Manag Sci 58: 876-888.
Bardas GA, Myresiotis CK, Karaoglanidis GS (2008) Stability and fitness of anilinopyrimidine-resistant strains of Botrytis cinerea. Phytopatholy 98: 443-450.
Kretschmer M, Leroch M, Mosbach A, Walker AS, Fillinger S, Mernke D, Schoonbeek HJ, Pradier JM, Leroux P, De Waard MA, Hahn M (2009) Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea. PLoS Pathog 5: el 000696.
Kovach J, Petzoldt R, Harman GE (2000) Use of honeybees and bumble bees to disseminate Trichoderma harzianum 1295-22 to strawberries for Botrytis control. Biol Control 18: 235-

Claims

1. A method for disseminating chemical plant protection agents in agriculture and horticulture, said method comprising the use of pollinators.
2. The method according to claim 1, further comprising the use of a beehive dispenser.
3. The method according to claim 2, wherein the beehive dispenser is mounted to the exit of the beehive.
4. Use of pollinators to disseminate chemical plant protection agents in agriculture and horticulture.
5. The method according to any one of claims 1 to 3; or the use according to claim 4, wherein the pollinators are bumblebees.
6. The method according to any one of claims 1 to 3, or 5; or the use according to claim 4, wherein the chemical plant protection agents are insecticides or fungicides.
7. The method according to any one of claims 1 to 3, or 5 to 6; or the use according to claim 4, wherein the chemical plant protection agents are fungicides, in particular botryticides .
8. The method according to any one of claims 1 to 3, or 5 to 7; or the use according to claim 4, wherein the chemical plant protection agents are used at their maximum field recommended concentration.
9. The method according to any one of claims 1 to 3, or 5 to 7; or the use according to claim 4, wherein the chemical plant protection agents are used in combination with a suitable carrier; in particular selected from the group consisting of a starch such as corn or potato starch; talc; dust derived from crushed hulls of nuts; clay dust; or mixtures thereof.
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9357752B2 (en) 2011-04-07 2016-06-07 Bee Vectoring Technology Inc. Apparatus for treatment of plants
US9380777B2 (en) 2012-03-12 2016-07-05 Bee Vectoring Technology Inc. Formulation comprising a particulate calcium silicate and clonostachys rosea for treating plants
US9526233B2 (en) 2013-02-01 2016-12-27 Bee Vectoring Technology Inc. Apparatus for treatment of plants
WO2019201777A1 (en) 2018-04-19 2019-10-24 Evonik Degussa Gmbh Biocompatible siloxanes for formulation of microorganisms
WO2020234035A1 (en) 2019-05-20 2020-11-26 Evonik Operations Gmbh Use of polyglycerol esters als as carriers for microbiological active ingredients
WO2021219505A1 (en) 2020-04-30 2021-11-04 Evonik Operations Gmbh Use of polyethers as carriers for active ingredients

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5348511A (en) * 1993-05-12 1994-09-20 The United States Of America As Represented By The Secretary Of Agriculture Beehive-mounted device for utilizing honeybees (hymenoptera: apidae) in the dissemination of biocontrol agents
US5989100A (en) * 1997-09-08 1999-11-23 Cornell Research Foundation, Inc. Bee disseminator of biological controls
WO2002094014A1 (en) * 2001-05-22 2002-11-28 Amberley Management Services Limited Method and apparatus for distributing treatment agents
WO2006120431A2 (en) * 2005-05-12 2006-11-16 Exosect Limited A method of delivering a biologically active agent

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5348511A (en) * 1993-05-12 1994-09-20 The United States Of America As Represented By The Secretary Of Agriculture Beehive-mounted device for utilizing honeybees (hymenoptera: apidae) in the dissemination of biocontrol agents
US5989100A (en) * 1997-09-08 1999-11-23 Cornell Research Foundation, Inc. Bee disseminator of biological controls
WO2002094014A1 (en) * 2001-05-22 2002-11-28 Amberley Management Services Limited Method and apparatus for distributing treatment agents
WO2006120431A2 (en) * 2005-05-12 2006-11-16 Exosect Limited A method of delivering a biologically active agent

Non-Patent Citations (24)

* Cited by examiner, † Cited by third party
Title
AL- MAZRA'AWI ET AL., BIOLOGICAL CONTROL, vol. 36, 2007, pages 89 - 97
AL-MAZRA'AWI ET AL., ENVIRON. ENTOMOL., vol. 35, no. 6, 2006, pages 1569 - 1577
ANA R. CABRERA; RAYMOND A. CLOYD; EDMOND R. ZABORSKI: "Effects of Greenhouse Pesticides on the Soil-Dwelling Predatory Mite Stratiolaelaps scimitus (Acari: Mesostigmata: Laelapidae) Under Laboratory Conditions", JOURNAL OF ECONOMIC ENTOMOLOGY, vol. 97, no. 3, 2004, pages 793 - 799
BARDAS GA; MYRESIOTIS CK; KARAOGLANIDIS GS: "Stability and fitness of anilinopyrimidine-resistant strains of Botrytis cinerea", PHYTOPATHOLY, vol. 98, 2008, pages 443 - 450
DESNEUX ET AL., ANNUAL REVIEW OF ENTOMOLOGY, vol. 52, 2007, pages 81 - 106
DIANEZ F; SANTOS M; BLANCO R; TELLO JC: "Fungicide resistance in Botrytis cinerea isolates from strawberry crops in Huelva (southwestern Spain)", PHYTOPARASITICA, vol. 30, 2002, pages 529 - 534
GRADISH AE; SCOTT-DUPREE DC; SHIPP L; HARRIS CR; FERGUSON G: "Effect of reduced risk pesticides for use in greenhouse vegetable on Bombus impatiens (Hymenoptera: Apidae)", PEST MANAG SCI, vol. 66, 2010, pages 142 - 146
HEINRICH B: "Bumblebee Economics", 1979, HARVARD UNIVERSITY PRESS, pages: 245
KAPONGO ET AL., BIOCONTROL, vol. 53, 2008, pages 797 - 812
KAPONGO ET AL., BIOLOGICAL CONTROL, 2008, pages 508 - 514
KOVACH ET AL., BIOLOGICAL CONTROL, vol. 18, 2000, pages 235 - 242
KOVACH J; PETZOLDT R; HARMAN GE: "Use of honeybees and bumble bees to disseminate Trichoderma harzianum 1295-22 to strawberries for Botrytis control", BIOL CONTROL, vol. 18, 2000, pages 235 - 242
KRETSCHMER M; LEROCH M; MOSBACH A; WALKER AS; FILLINGER S; MERNKE D; SCHOONBEEK HJ; PRADIER JM; LEROUX P; DE WAARD MA: "Fungicide-driven evolution and molecular basis ofmultidrug resistance in field populations of the grey mould fungus Botrytis cinerea", PLOS PATHOG, vol. 5, 2009, pages 000696
LEROUX P; FRITZ R; DEBIEU D; ALBERTINI C; LANEN C; BACH J; GREDT M; CHAPELAND F: "Mechanisms of resistance to fungicides in field strains of Botrytis cinerea", PEST MANAG SCI, vol. 58, 2002, pages 876 - 888
MACCAGNANI ET AL., BULLETIN OF INSECTOLOGY, vol. 58, no. 1, 2005, pages 3 - 8
MALONE LA; SCOTT-DUPREE CD; TODD JH; RAMANKUTTY P: "No sublethal toxicity to bumblebees, Bombus terrestris, exposed to Bt-corn pollen, captan and novaluron", NZJ CROP HORTIC SCI, vol. 35, 2007, pages 435 - 439
NAKAMURA M; KONO Y; TAKATSUKI A: "Mepanipyrim, a novel inhibitor of pharmacologically induced Golgi dispersion", BIOSCI BIOTECHNOL BIOCHEM, vol. 67, 2003, pages 139 - 150
SHAFIR ET AL., EUR. J. OF PLANT PATHOLOGY, vol. 116, 2006, pages 119 - 128
SKERL MIS; BOLTA SV; CESNIK HB; GREGORC A: "Residues of pesticides in honeybee (Apis mellifera carnica) bee bread and in pollen loads from treated apple orchards", BULL ENVIRON CONTAM TOXICOL, vol. 83, 2009, pages 374 - 377
SMITH ET AL., PHYTOPATHOLOGY, vol. 99, no. 6, 2009, pages 121
SUTTON J C; PENG G: "Manipulation and Vectoring of Biocontrol Organisms to Manage Foliage and Fruit Diseases in Cropping Systems", ANNUAL REVIEW OF PHYTOPATHOLOGY, vol. 31, September 1993 (1993-09-01), Palo Alto, U.S.A., pages 473 - 493, XP002616272, ISSN: 0066-4286, DOI: 10.1146/annurev.py.31.090193.002353 *
SUTTON; PENG, ANNUAL REVIEW OF PHYTOPATHOLOGY, vol. 31, 1993, pages 473 - 493
TERADA M; MIZUHASHI F; MURATA K; TOMITA T: "Mepanipyrim, a new fungicide, inhibits intracellular transport of very low density lipoprotein in rat hepatocytes", TOXICOL APPL PHARMACOL, vol. 154, 1999, pages 1 - 11
WALTER M; OBANOR F O; DONOVAN B; BOYD-WILSON K S H; NEAL M; SIEFKES-BOER H J; LANGFORD G I: "Dissemination of biological and chemical fungicides by bees onto Rubus and Ribes flowers", APPS 2009: PLANT HEALTH MANAGEMENT: AN INTEGRATED APPROACH, September 2009 (2009-09-01), pages 48, XP002616273, ISBN: 9780646529196, Retrieved from the Internet <URL:http://www.australasianplantpathologysociety.org.au/Publications/Proceedings/APPS%202009%20Handbook%20(revised)%20(09Feb10).pdf> [retrieved on 20110111] *

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US9968099B2 (en) 2012-03-12 2018-05-15 Bee Vectoring Technology Inc. Formulation comprising a particulate calcium silicate and clonostachys rosea for treating plants
US9526233B2 (en) 2013-02-01 2016-12-27 Bee Vectoring Technology Inc. Apparatus for treatment of plants
WO2019201777A1 (en) 2018-04-19 2019-10-24 Evonik Degussa Gmbh Biocompatible siloxanes for formulation of microorganisms
WO2020234035A1 (en) 2019-05-20 2020-11-26 Evonik Operations Gmbh Use of polyglycerol esters als as carriers for microbiological active ingredients
WO2021219505A1 (en) 2020-04-30 2021-11-04 Evonik Operations Gmbh Use of polyethers as carriers for active ingredients

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