WO2023170145A1 - Mite composition and method for feeding predatory mites - Google Patents

Mite composition and method for feeding predatory mites Download PDF

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Publication number
WO2023170145A1
WO2023170145A1 PCT/EP2023/055889 EP2023055889W WO2023170145A1 WO 2023170145 A1 WO2023170145 A1 WO 2023170145A1 EP 2023055889 W EP2023055889 W EP 2023055889W WO 2023170145 A1 WO2023170145 A1 WO 2023170145A1
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Prior art keywords
mites
mite
predatory
composition
prey
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PCT/EP2023/055889
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French (fr)
Inventor
Clive S.A. STINSON
Pavel B. KLIMOV
Emile P. DEMARD
Jared L. DUNHAM
Marcus Vinicius Alfenas DUARTE
Dominiek VANGANSBEKE
Karel Jozef Florent Bolckmans
Felix Leopold WÄCKERS
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Biobest Group N.V.
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Publication of WO2023170145A1 publication Critical patent/WO2023170145A1/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
    • 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
    • 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
    • A01N63/16—Arachnids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/02—Acaricides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/04—Insecticides

Definitions

  • the present invention relates to the use of prey mites from the species Thyreophagus calusorum as a food for predatory mites, which can be used for controlling a pest in a crop.
  • Predatory mites are traditionally used in the broad field of agricultural pest management.
  • a wide range of predatory mite species have been suggested or commercialized for the biological control of phytophagous pest mites and insect pests such as whiteflies and thrips.
  • Biocontrol is the intentional manipulation of populations of living beneficial organisms (natural enemies) in order to limit population of pests. Natural enemies of mites for instance include predators, parasitic insects, nematodes, and pathogens. Indeed, virtually all pests have natural enemies and appropriate management of such natural enemies can effectively control many pests.
  • biological control is not to eradicate pests, but to maintain them at tolerable levels at which they cause no appreciable damage. As such, biological control can be effective, economical and safe.
  • predatory mite populations have been used in order to control pests.
  • phytoseiid predatory mites are currently used to control pests such as phytophagous mites, thrips and whiteflies.
  • Neoseiulus cucumeris (Oudemans) is commercially used for the control of thrips larvae and spider mites.
  • Other predatory mites species particularly mesostigmatid and prostigmatid species, receive particular attention in the context of biological pest control, and some are already commercialized.
  • W02006/057552, W02008/015393, W02008/104807 and W02007/075081 demonstrate the potential of astigmatid mites to be used as prey mites in mass-rearing of predatory mites.
  • Astigmatid prey mite species are used in mass rearing methods, i.e. Tyrophagus putrescentiae (Schrank), Thyreophagus entomophagus (Laboulbene & Robin) and Carpoglyphus lactis L..
  • W02008/015393 A2 discloses comparative trials between these three commercial prey mites and found that Thyreophagus entomophagus outperforms both T. putrescentiae and C. lactis.
  • the inventors have surprisingly found that the use of mites from the species Thyreophagus calusorum as prey mites overcome the problems of the prior art.
  • the present invention provides a mite composition
  • a mite composition comprising
  • said prey mite population comprises mites from the species Thyreophagus calusorum.
  • Fig. 1 Number of eggs deposited by N. californicus when fed with Th. entomophagus or Th. calusorum (means ⁇ standard error).
  • Fig. 2 Number of eggs and mobiles of A. swirskii when fed with Th. entomophagus or Th. calusorum (means ⁇ standard error).
  • Fig. 3 Number of eggs and mobiles of N. californicus when fed with Th. entomophagus or Th. calusorum (means ⁇ standard error).
  • Fig. 4 Egg-to-mobile ratio for C. transversostriata and Th. calusorum (means ⁇ standard error).
  • the present invention provides a mite composition
  • a mite composition comprising
  • said prey mite population comprises mites from the species Thyreophagus calusorum.
  • predatory mite population » is meant in the sense of the present invention, a population of beneficial mites that feeds on a prey mite population.
  • prey mite population » is meant in the sense of the present invention, a population of mites that at least partially consumed by a predatory mite population.
  • carrier » is meant in the sense of the present invention, any solid material which is suitable to provide a carrier surface to the individuals of both the predatory and the prey mite populations.
  • the carrier will usually act as a three-dimensional matrix wherein the prey mite population and predatory mite population can move around, hide, develop and prey or be preyed upon.
  • Thyreophagus calusorum is parthenogenetically reproducing astigmatid mite which was collected from subcortical habitats in Florida, USA.
  • T. calusorum females differ from all known species of Thyreophagus species by having very short and somewhat conical solenidion o on tarsus II, spermathecae which are broad, sclerotized and arc-shaped at the entrance of the spermathecal canal and by the absence of ventral seta wa l-ll (Klimov et al. submitted).
  • the composition according to the invention comprises a food source for the predatory and/or prey mite populations.
  • the composition of the invention comprises a food source for Thyreophagus calusorum.
  • the food source is a natural diet for Thyreophagus calusorum.
  • the food source may be derived from grain.
  • the food source comprises wheat germ, bran, rice hulls, rolled oats, corn grits, flour (such as gram flour, buckwheat flour or cereal flour), dried fruit, jam, dried insects, fungi (such as Saccharomyces cerevisiae) or poultry meal.
  • Mite diets can be supplemented with sugars, such as sucrose, glucose, fructose or dextrose.
  • the food source is an artificial diet.
  • the diet may additionally contain other food sources that are desirable for the predator and/or prey mite to feed on.
  • the diet may contain pollen such as that from bee-collected pollen, Typha sp., date palm plant Phoenix dactylifera, or castor oil plant Ricinus communis.
  • the food source comprises bee pollen.
  • a carrier can be selected that acts both as a carrier and as a food source, for example wheat germ.
  • the composition according to the invention may comprise a fungus.
  • the fungus is a yeast, preferably a Saccharomycetes, such as from the genus Saccharomyces.
  • the fungus is Saccharomyces cerevisiae. It has even been found that a fungus, such as yeast, is a sufficient food source for the prey mites of the invention. As a consequence, rearing of the prey mites of the invention is more practical, stable and economical than the rearing of several other prey mites that are currently used or suggested for the rearing of predatory mites.
  • a supplemental food source as used herein refers to a food source that is present in addition to the prey mite population. Supplemental food sources are sometimes used in predatory mite rearing as a food source that is more cost-effective than prey mites. It is often observed that several predatory mites can use supplemental foods for obtaining additional energy, but also require the presence of prey mites for survival, development and reproduction. A major drawback of the use of a supplemental food source is that it is prone to fungal contamination. By combining a supplemental food source with the prey mites of the invention, this drawback is suppressed. Examples of preferred supplemental foods are natural food sources (such as pollen or dead mites) and artificial foods (Wackers et al.
  • the prey mite composition according to the invention can comprise (or consist of) a population of immobilized and/or dead prey mites and/or dead prey mite eggs.
  • the prey mite composition according to the invention comprises dead prey mites and dead prey mite eggs.
  • the prey mite population as used in the invention comprises (a) a population of dead prey mites, and (b) a population of living mites of the species Thyreophagus calusorum.
  • the prey mite population as used in the invention comprises (a) a population of dead prey mites from the species Thyreophagus calusorum, and (b) a population of living mites from the species Thyreophagus calusorum.
  • the prey mite population comprising mites from the species Thyreophagus calusorum has been treated by a treatment selected from the group consisting of: thermal treatment, such as freezing, heating, cold-shock or heat-shock treatment; chemical treatment, such as gas or fume treatment; radiation treatment, such as UV, microwave, gamma irradiation or X-ray treatment; mechanical treatment, such as vigorous shaking, or stirring, subjecting to shear forces, collision; gas pressure treatment, such as ultrasound treatment, pressure changes, pressure drops; electrical treatment, such as electrocution; immobilizing with an adhesive; immobilization by starvation, such as induced by water or food deprivation; immobilization by suffocation or anoxia treatment, such as by temporarily eliminating oxygen from the atmosphere or replacing oxygen by another gas and any combination thereof.
  • thermal treatment such as freezing, heating, cold-shock or heat-shock treatment
  • chemical treatment such as gas or fume treatment
  • radiation treatment such as UV, microwave, gamma irradiation or
  • the prey mite population comprising mites from the species Thyreophagus calusorum has been killed, for example by freezing or irradiation.
  • the composition of the invention comprises mites from the species Thyreophagus calusorum that have been killed by freezing. It is therefore also an aspect of the invention to provide a method for rearing predatory mites, the method comprising
  • the killed prey mite population comprising mites from the species Thyreophagus calusorum with a predatory mite population and a carrier for individuals of the mite populations, and
  • the carrier can be any solid material, which is suitable to provide a carrier surface to the individuals.
  • the carrier is a mixture of solid carrier elements.
  • the carrier is a substantially homogeneous mixture of solid carrier elements. While the solid carrier elements may have a variety of sizes, a substantially homogeneous mixture refers to the variety of sizes being substantially homogeneously distributed amongst the carrier.
  • the average longest axis of the solid carrier elements is typically between 1 .0 and 20.0 mm, more in particular between 2.0 and 12.0 mm, preferably between 3.0 and 9.0 mm.
  • the carrier is a mixture of solid carrier elements with a similar size.
  • 90% of the solid carrier elements have a longest axis in the range of 0.1 times to 10.0 times the average longest axis of the mixture of solid carrier elements, in particular in the range of 0.2 and 5.0 times, more in particular in the range of 0.5 and 2.0 times.
  • two or more different types of carrier elements may be used and mixed. In such case, the size distribution refers to the size distribution per type of carrier elements.
  • the carrier provides a porous medium, which allows exchanges of metabolic gases and heat produced by the mite populations.
  • suitable carriers are non-photosynthetic plant material, such as (wheat) bran, buckwheat husks, rice husks, saw dust, tree bark, corn cob grits, etcetera, or inorganic material, such as vermiculite.
  • said carrier comprises grains of a grass species or any part thereof, such as germ or bran.
  • said carrier does not comprise living, green plant material, such as leaves or stems of plants.
  • the carrier comprises non-photosynthetic plant material.
  • a carrier selected from vermiculite, wheat bran, millet chaff, rice husks and buckwheat husks.
  • said carrier comprises carrier elements with an average longest axis of between 1.0 and 20.0 mm, in particular between 3.0 and 9.0 mm.
  • the carrier material may provide shelter for the prey mites and predatory mites, reducing stress conditions and cannibalism.
  • the carrier allows for the generation of a three-dimensional rearing matrix and may provide a food source for the prey mites.
  • the carrier acts as a bulking agent and allows for a more homogeneous distribution of the predatory mites in the crops.
  • the number of individuals of the predatory mite population relative to the number of individuals of the prey mite population is from about 5:1 to 1 :500, such as about 2:1 to 1 :250, and preferably between 1 :1 and 1 :150.
  • the number of individuals of the predatory mite population relative to the number of individuals of the prey mite population is from about 1 :5 to 1 :30.
  • the mite compositions of the present invention comprise at least 1000 mites of the prey mite population per gram of the composition, in particular at least 2000 prey mites, more in particular at least 3000 prey mites. In a further embodiment, at least 5000 prey mites, at least 10,000 prey mites, at least 20,000 prey mites, at least 30,000 prey mites. In a preferred embodiment, the mite composition comprises at least 50,000 mites of the prey mite population per gram of the composition.
  • the mite compositions of the present invention comprise at least 20 mites of the predatory mite population per gram of the composition, in particular at least 30 predatory mites, more in particular at least 50 predatory mites. In a further embodiment, at least 100 predatory mites, at least 150 predatory mites, at least 200 predatory mites, at least 300 predatory mites. In a preferred embodiment, the mite composition comprises at least 500 mites of the predatory mite population per gram of the composition.
  • Mite numbers are mentioned herein refer to the total number of all mite development stages, thus including eggs, larvae, nymphs, all chrysalis stages and adults, unless the context clearly dictates otherwise.
  • the prey mites in the compositions of the invention are not required to be living prey mites.
  • the mite compositions of the present invention comprise at least 1000 non-egg mites of the prey mite population per gram of the composition, in particular at least 2000 non-egg prey mites, more in particular at least 3000 non-egg prey mites. In a further embodiment, at least 5000 prey non-egg mites, at least 10,000 prey non-egg mites, at least 20,000 prey non-egg mites, at least 30,000 prey non-egg mites. In a preferred embodiment, the mite composition comprises at least 50,000 non-egg mites of the prey mite population per gram of the composition.
  • the mite compositions of the present invention comprise at least 20 non-egg mites of the predatory mite population per gram of the composition, in particular at least 30 non-egg predatory mites, more in particular at least non- egg 50 predatory mites. In a further embodiment, at least 100 non-egg predatory mites, at least 150 non-egg predatory mites, at least 200 non-egg predatory mites, at least 300 non-egg predatory mites. In a preferred embodiment, the mite composition comprises at least 500 non- egg mites of the predatory mite population per gram of the composition.
  • the mite compositions of the present invention comprise at least 20 adult mites of the predatory mite population per gram of the composition, in particular at least 30 adult predatory mites, more in particular at least 50 adult predatory mites. In a further embodiment, at least 100 adult predatory mites, at least 150 adult predatory mites, at least 200 adult predatory mites, at least 300 adult predatory mites. In a preferred embodiment, the mite composition comprises at least 500 adult mites of the predatory mite population per gram of the composition.
  • the mite compositions of the present invention comprise at least 20 living adult mites of the predatory mite population per gram of the composition, in particular at least 30 living adult predatory mites, more in particular at least 50 living adult predatory mites. In a further embodiment, at least 100 living adult predatory mites, at least 150 living adult predatory mites, at least 200 living adult predatory mites, at least 300 living adult predatory mites. In a preferred embodiment, the mite composition comprises at least 500 living adult mites of the predatory mite population per gram of the composition.
  • the present invention provides, for example, mite compositions comprising at least 1000 mites of the prey mite population and at least 20 mites of the predatory mite population per gram of the composition. As another example, at least 5000 mites of the prey mite population and at least 100 mites of the predatory mite population. As another example, at least 50,000 mites of the prey mite population and at least 500 mites of the predatory mite population.
  • said predatory mites are mesostigmatid or prostigmatid mite species. In a further embodiment, said predatory mites are selected from:
  • Phytoseiidae such as from: a)- the subfamily of the Amblyseiinae, such as from the genus Amblyseius, e.g. Amblyseius andersoni, Amblyseius aerialis, Amblyseius swirskii, Amblyseius herbicolus or Amblyseius largoensis, from the genus Euseius e.g.
  • Euseius finlandicus Euseius h ibisci, Euseius ovalis, Euseius victoriensis, Euseius stipulatus, Euseius scutalis, Euseius tularensis, Euseius addoensis, Euseius concordis, Euseius ho or Euseius citri, from the genus Neoseiulus e.g.
  • Neosei ulus barkeri Neoseiulus califomicus, Neoseiulus cucumeris, Neoseiulus longispinosus, Neoseiulus womersleyi, Neoseiulus idaeus, Neoseiulus anonymus, Neoseiulus paspalivorus, Neoseiulus reductus or Neoseiulus fallacis, from the genus Amblydromalus e.g. Amblydromalus limonicus from the genus Typhlodromalus e.g.
  • Typhlodromalus aripo, Typhlodromalus laila or Typhlodromalus peregrinus, from the genus Transeius e.g. Transeius montdorensis, from the genus Phytoseiulus, e.g. Phytoseiulus persimilis, Phytoseiulus macropilis, Phytoseiulus longipes, Phytoseiulus fragariae; b) the subfamily of the Typhlodrominae, such as from the genus Galendromus e.g. Galendromus occidentalis from the genus Typhlodromus e.g.
  • Typhlodromus pyri Typhlodromus doreenae, Typhlodromus rhenanus or Typhlodromus athiasae
  • Phytoseiinae such as from the genus Phytoseius e.g. Phytoseius macropilis, Phytososeius finitimus or Phytoseius plumifer.
  • Ascidae such as from the genus Proctolaelaps, such as Proctolaelaps pygmaeus (Muller); from the genus Blattisocius e.g.
  • Blattisocius tarsalis (Berlese), Blattisocius keegani (Fox); from the genus Lasioseius e.g. Lasioseius fimetorum Karg, Lasioseius floridensis Berlese, Lasioseius bispinosus Evans, Lasioseius dentatus Fox, Lasioseius scapulatus (Kenett), Lasioseius athiasae Nawar & Nasr; from the genus Arctoseius e.g. Arctoseius semiscissus (Berlese); from the genus Protogamasellus e.g.
  • Lasioseius fimetorum Karg Lasioseius floridensis Berlese, Lasioseius bispinosus Evans, Lasioseius dentatus Fox, Lasiosei
  • Laelapidae such as from the genus Stratiolaelaps e.g. Stratiolaelaps scimitus (Womersley) (also placed in the genus Hypoaspis) Gaeolaelaps e.g. Gaeolaelaps aculeifer (Canestrini) (also placed in the genus Hypoaspis); Androlaelaps e.g. Androlaelaps casalis (Berlese); iv) Macrochelidae such as from the genus Macrocheles e.g.
  • Macrocheles robustulus (Berlese), Macrocheles muscaedomesticae (Scopoli), Macrocheles matrius (Hull);
  • Parasitidae such as from the genus Pergamasus; e.g. Pergamasus quisquiliarum Canestrini; Parasitus e.g. Parasitusfimetorum (Berlese), Parasitus bituberosus Karg; Parasitellus e.g. Parasitellus fucorum (De Geer);
  • Digamasellidae such as from the genus Digamasellus; e.g. Digamasellus quadrisetus or Digamasellus punctum; from the genus Dendrolaelaps; e.g. Dendrolaelaps neodisetus; and
  • -prostigmatid mite species such as from: i) Tydeidae such as from the genus Homeopronematus e.g. Homeopronematus anconai (Baker); from the genus Tydeus e.g. Tydeus Iambi (Baker), Tydeus caudatus (Duges); from the genus Pronematus e.g. Pronematus ubiquitus (McGregor); ii) Cheyletidae such as from the genus Cheyletus e.g.
  • Cheyletus eruditus (Schrank Cheyletus malaccensis Oudemans; iii) Cunaxidae such as from the genus Coleoscirus e.g. Coleoscirus simplex (Ewing), from the genus Cunaxa e.g. Cunaxa setirostris (Rermann); iv) Erythraeidae such as from the genus Balaustium e.g. Balaustium putmani Smiley, Balaustium medicagoense Meyer & Ryke , Balaustium murorum (Hermann); v) Stigmaeidae such as from the genus Agistemus e.g.
  • Agistemus exsertus Gonzalez such as from the genus Zetzellia e.g. Zetzellia mali (Ewing); and vi) Tarsonemidae such as from the genus Acaronemus, e.g. Acaronemus destructor (Smiley and Landwehr); such as from the genus Dendroptus near suski Sharonov and Livshits; such as Lupotarsonemus floridanus Attiah.
  • the predatory mites belong to the Phytoseiidae.
  • the predatory mites are selected from Amblyseiinae, Transeius, Neoseiulus and Amblydromalus; more in particular the predatory mites are selected from the group consisting of Amblyseius swirskii, Transeius montdorensis, Neoseiulus californicus, Amblydromalus limonicus, Neoseiulus cucumeris, Amblyseius andersoni, Iphiseius degenerans and Stratiolaelaps scimitus; more in particular from the group consisting of Amblyseius swirskii, Transeius montdorensis, Neoseiulus californicus, and Amblydromalus limonicus.
  • the predatory mites are selected from Amblyseiinae, Transeius, Neo
  • the present invention also provides a commercial packaging for storing and distributing the composition of the invention. Therefore, the present invention further pertains to a container comprising a composition according to the invention.
  • the container has an internal volume of between 0.004 I and 5 I, preferably between 0.5 I and 2 I.
  • the container preferably comprising an exit for at least one motile life stage of the mite, more preferably an exit suitable for providing a sustained release of said at least one motile life stage.
  • the container e.g. a bottle or a sachet
  • seal as used herein refers to a closure that is glued or heat-sealed.
  • the container is adapted to attach it to a crop, for example by comprising a hook to hang it from a crop leaf or branch or by comprising a sticky surface to stick it to a surface of the crop or with a stick attached to the sachet.
  • the present invention further pertains to a method for rearing predatory mites, the method comprising
  • the individuals of the predatory mite population are allowed to feed on said prey mite population while maintaining said mite composition at 5 to 35 °C and 30 to 100% relative humidity; in particular at 15 to 35°C and 50 to 100% relative humidity.
  • the present invention further pertains to a method for controlling a pest in a crop, the method comprising providing to said crop a mite composition according to the invention.
  • the mite composition of the invention may be distributed directly onto the crop.
  • the mite composition of the invention is provided in the proximity of the crop.
  • the mite composition of the invention may be provided to a crop by placing a container holding the mite composition in the vicinity of the crop and allowing the predatory mites to exit from said container. This way, predatory mites spread themselves through the crops and control the crop pests.
  • the pest is an arthropod pest.
  • the present invention further provides the use of mites from the species Thyreophagus calusorum for feeding predatory mites. Conveniently, it has been found that these mites can be reared efficiently and provide an easy food source for predatory mites.
  • the present application allows to distribute mites from the species Thyreophagus calusorum in a crop such that predatory mites that are present in the crop can feed thereon.
  • the mites from the species Thyreophagus calusorum (herein also referred to as prey mites or prey mite population) may be distributed on their own or may be distributed in combination with predatory mites. When distributed in combination with predatory mites, the predatory mites and prey mites may be distributed at the same time or at distinct times.
  • Example 1 Reproduction of Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites
  • Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female N. californicus were individually transferred from the mass-rearing to a PVC arena (1 x 2 cm) of which the edges were covered with tissue paper. A small volume of prey mites (0.1 mL) was provided as food for the females. The trial was conducted in a climatic chamber set at 23 ⁇ 1 °C, 90 ⁇ 5 % and a 16:8 h (L:D) photoperiod.
  • Results were analyzed by counting and removing the deposited eggs on a daily basis. Data of the first day was omitted to reduce the effects of pre-experimental conditions (van Houten et al. 1995). The number of eggs and mobiles was compared using a generalized linear model with a Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014). As demonstrated in Fig. 1 , a significantly higher number of eggs were deposited when N. californicus was fed with Th. calusorum compared to Th. entomophagus (GLM: p ⁇ 0.05). It can hence be concluded that Th. calusorum is more efficacious prey mite for rearing predatory mites.
  • Example 2 Growth comparison of predatory mite Amblyseius swirskii on Th. entomophagus and Th. calusorum prey mites
  • This experiment was conducted to compare the population growth of the phytoseiid predatory mite Amblyseius swirskii on two species of astigmatid prey mites, namely Thyreophagus entomophagus and Thyreophagus calusorum. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female A. swirskii were transferred to an experimental breeding dish (5 x 1.5 cm) (SPL Life Sciences, Korea), which was filled with sawdust. A small volume of prey mites (0.5 mL) was added to the mix as food for the predatory mites.
  • the trial was conducted in a climatic chamber set at 23 ⁇ 1 °C, 90 ⁇ 5 % and a 16:8 h (L:D) photoperiod. Results were analyzed by comparing the number of predatory mite mobiles and eggs after 10 days using a generalized linear model with a quasi Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014).
  • Th. calusorum As shown in Fig. 2, the number of eggs and mobiles of A. swirskii fed on Th. entomophagus were highly similar to those fed on Th. calusorum (GLM: p>0.05 for both eggs and mobiles). This signifies the suitability of using Th. calusorum as prey mite for rearing predatory mites.
  • Example 3 Growth comparison of predatory mite Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites
  • This experiment was conducted to compare the population growth of the phytoseiid predatory mite Neoseiulus californicus on two species of astigmatid prey mites, namely Th. entomophagus and Th. calusorum. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female N. californicus were transferred to an experimental breeding dish (5 x 1 .5 cm) (SPL Life Sciences, Korea), which was filled with sawdust. A small volume of prey mites (0.5 mL) was added to the mix as food for the predatory mites.
  • the trial was conducted in a climatic chamber set at 23 ⁇ 1 °C, 90 ⁇ 5 % and a 16:8 h (L:D) photoperiod. Results were analyzed by comparing the number of predatory mite mobiles and eggs after 10 days using a generalized linear model with a quasi Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014).
  • Example 4 Comparison of egg/mobile ratio between C. transversostriata and Th. calusorum prey mites
  • a prey mite For feeding phytoseiid predatory mites, a prey mite should preferably have a high proportion of eggs.
  • the capture success-rate is higher in egg stages as compared to nymphal or adults stages, also referred to as mobiles (Midthassel et al. 2013).
  • This experiment was conducted to compare the proportion of eggs to mobiles between two species of astigmatid prey mites, namely Czenspinskia transversostriata and Th. calusorum, the prey mite of the invention. Random production units containing 1 liter of volume were selected from the massproduction of both Th. calusorum and C. transversostriata. A subsample was taken and the number of egg and mobiles stages were counted separately under a stereo microscope. The egg/mobile ratio of both species was compared with a generalized linear model using the statistical software R (version 3.6.1 ) (R Core Team 2014).
  • Th. calusorum prey mites are a more efficient food source for phytoseiid predatory mites, compared with C. transversostriata.

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Abstract

The present invention relates to the use of prey mites from the species Thyreophagus calusorum as a food for predatory mites, which can be used for controlling a pest in a crop.

Description

Mite composition and method for feeding predatory mites
Field of the invention
The present invention relates to the use of prey mites from the species Thyreophagus calusorum as a food for predatory mites, which can be used for controlling a pest in a crop.
Background of the invention
Predatory mites are traditionally used in the broad field of agricultural pest management. A wide range of predatory mite species have been suggested or commercialized for the biological control of phytophagous pest mites and insect pests such as whiteflies and thrips.
Lately, the major arthropod pest species have mainly been controlled by chemical agents, i.e. pesticides. Nowadays, many of the new pesticides made available on the market are more selective and less hazardous than the older compounds. However, even the newest pesticides present several major problems, i.e. the development of resistance in target pest species; the dwindling supply of useful, registered insecticides and acaricides; the deposition of undesirable residues; the detrimental effect on non-target species resulting in secondary pest outbreaks; the phytotoxic reactions induced in treated plants. It is therefore becoming increasingly clear that solely relying on chemical control will not be the solution to the problem of agricultural pest management. For this reason, many farmers and gardeners are exploring and adopting methods to reduce pesticide use.
One alternative to the use of chemical agents is biological control. Biological control is the intentional manipulation of populations of living beneficial organisms (natural enemies) in order to limit population of pests. Natural enemies of mites for instance include predators, parasitic insects, nematodes, and pathogens. Indeed, virtually all pests have natural enemies and appropriate management of such natural enemies can effectively control many pests.
The objective of biological control is not to eradicate pests, but to maintain them at tolerable levels at which they cause no appreciable damage. As such, biological control can be effective, economical and safe.
Since many years, predatory mite populations have been used in order to control pests. In particular, primarily phytoseiid predatory mites are currently used to control pests such as phytophagous mites, thrips and whiteflies. For example, Neoseiulus cucumeris (Oudemans) is commercially used for the control of thrips larvae and spider mites. Other predatory mites species, particularly mesostigmatid and prostigmatid species, receive particular attention in the context of biological pest control, and some are already commercialized.
There is therefore a growing interest for the development of effective mass rearing systems to produce predatory mite populations on commercially-relevant scales, and this for an acceptable price. Currently, commercial rearing systems use living preys in a culture maintained on a carrier to rear predatory mites. For example, W02006/071 107 and WO2013/103295 disclose a mite composition comprising a population of individuals of a predatory mite species, a prey mite population as a food source for the predatory mite individuals and a carrier. The compositions according to these prior art documents are suitable for rearing mite species and for the biological control of pests. Mass rearing systems for predatory mites depend heavily on the availability of suitable prey for the predators. Considering their role in rearing of predatory mites, the commercial relevance of rearing prey mites is increasing.
W02006/057552, W02008/015393, W02008/104807 and W02007/075081 demonstrate the potential of astigmatid mites to be used as prey mites in mass-rearing of predatory mites. However, until now, only a few astigmatid prey mite species are used in mass rearing methods, i.e. Tyrophagus putrescentiae (Schrank), Thyreophagus entomophagus (Laboulbene & Robin) and Carpoglyphus lactis L.. W02008/015393 A2 discloses comparative trials between these three commercial prey mites and found that Thyreophagus entomophagus outperforms both T. putrescentiae and C. lactis.
Identifying new prey mites for use in biological pest control, which are at the same time economical and easy to breed remains a challenge. Most mites that are preyed upon by predatory mites in nature have subsequently been found not to be suitable for mass-rearing of these predatory mites, for example because they do not reach sufficiently high densities or have defense mechanisms against predatory mites (see e.g. Massaro, M., Martin, J.P.I. and de Moraes G.J. Exp Appl Acarol (2016) 70: 41 1 ; Barbosa, M.F.C. and de Moraes, G.J. Exp Appl Acarol (2016) 69: 289; Barbosa, M.F.C. and de Moraes, G.J. Biological Control (2015) 91 : 22-26; and Midthassel, A., Leather, S.R., Wright, D.J. et al. BioControl (2016) 61 : 437). Furthermore, any potential negative impact of the prey mite on crop plants as well as end users should be minimal.
In view of the above there is a continuing need to obtain alternative and, preferably improved (more efficient) mite compositions for mass rearing and large-scale production of predatory mite populations, for commercial distribution of larger volumes of mite compositions comprising such and for better pest control and agricultural management. Summary of the invention
The inventors have surprisingly found that the use of mites from the species Thyreophagus calusorum as prey mites overcome the problems of the prior art.
Therefore the present invention provides a mite composition comprising
- a predatory mite population,
- a prey mite population, and
- a carrier for individuals of said populations, wherein said prey mite population comprises mites from the species Thyreophagus calusorum.
Brief description of the drawings
Fig. 1 : Number of eggs deposited by N. californicus when fed with Th. entomophagus or Th. calusorum (means ± standard error).
Fig. 2: Number of eggs and mobiles of A. swirskii when fed with Th. entomophagus or Th. calusorum (means ± standard error).
Fig. 3: Number of eggs and mobiles of N. californicus when fed with Th. entomophagus or Th. calusorum (means ± standard error).
Fig. 4: Egg-to-mobile ratio for C. transversostriata and Th. calusorum (means ± standard error).
Detailed description of the invention
As described herein before, the present invention provides a mite composition comprising
- a predatory mite population,
- a prey mite population, and
- a carrier for individuals of said populations, wherein said prey mite population comprises mites from the species Thyreophagus calusorum.
By the term « predatory mite population », is meant in the sense of the present invention, a population of beneficial mites that feeds on a prey mite population.
By the term « prey mite population », is meant in the sense of the present invention, a population of mites that at least partially consumed by a predatory mite population. By the term « carrier », is meant in the sense of the present invention, any solid material which is suitable to provide a carrier surface to the individuals of both the predatory and the prey mite populations. The carrier will usually act as a three-dimensional matrix wherein the prey mite population and predatory mite population can move around, hide, develop and prey or be preyed upon.
The prey mite Thyreophagus calusorum is parthenogenetically reproducing astigmatid mite which was collected from subcortical habitats in Florida, USA. T. calusorum females differ from all known species of Thyreophagus species by having very short and somewhat conical solenidion o on tarsus II, spermathecae which are broad, sclerotized and arc-shaped at the entrance of the spermathecal canal and by the absence of ventral seta wa l-ll (Klimov et al. submitted).
Inventors surprisingly observed that a mite composition according to the invention shows better results in terms of efficiency. It was observed that individuals from a predatory mite population, which prey on mites of the species Thyreophagus calusorum, show a higher growth rate and therefore increase in the number of individuals in in the predatory mite composition.
Advantageously, the composition according to the invention comprises a food source for the predatory and/or prey mite populations. In a particular embodiment, the composition of the invention comprises a food source for Thyreophagus calusorum. In one embodiment, the food source is a natural diet for Thyreophagus calusorum. For example, the food source may be derived from grain. In one embodiment, the food source comprises wheat germ, bran, rice hulls, rolled oats, corn grits, flour (such as gram flour, buckwheat flour or cereal flour), dried fruit, jam, dried insects, fungi (such as Saccharomyces cerevisiae) or poultry meal. Mite diets can be supplemented with sugars, such as sucrose, glucose, fructose or dextrose. In another embodiment, the food source is an artificial diet. Optionally, the diet may additionally contain other food sources that are desirable for the predator and/or prey mite to feed on. For example, the diet may contain pollen such as that from bee-collected pollen, Typha sp., date palm plant Phoenix dactylifera, or castor oil plant Ricinus communis. In a particular embodiment, the food source comprises bee pollen. As is known to the skilled person, a carrier can be selected that acts both as a carrier and as a food source, for example wheat germ.
Advantageously, as explained herein before, the composition according to the invention may comprise a fungus. In a preferred embodiment, the fungus is a yeast, preferably a Saccharomycetes, such as from the genus Saccharomyces. In a preferred embodiment, the fungus is Saccharomyces cerevisiae. It has even been found that a fungus, such as yeast, is a sufficient food source for the prey mites of the invention. As a consequence, rearing of the prey mites of the invention is more practical, stable and economical than the rearing of several other prey mites that are currently used or suggested for the rearing of predatory mites. A supplemental food source as used herein refers to a food source that is present in addition to the prey mite population. Supplemental food sources are sometimes used in predatory mite rearing as a food source that is more cost-effective than prey mites. It is often observed that several predatory mites can use supplemental foods for obtaining additional energy, but also require the presence of prey mites for survival, development and reproduction. A major drawback of the use of a supplemental food source is that it is prone to fungal contamination. By combining a supplemental food source with the prey mites of the invention, this drawback is suppressed. Examples of preferred supplemental foods are natural food sources (such as pollen or dead mites) and artificial foods (Wackers et al. (2005) Plant- provided food for carnivorous insects: a protective mutualism and its applications 356p; Morales-Ramos et al. (2014) Mass production of beneficial organisms 742p; Cohen (2015) Insect diets. Science and Technology, Second edition, 473p) Supplemental food sources for the rearing of predatory mites are well-known to the skilled person.
In a particular embodiment, the prey mite composition according to the invention can comprise (or consist of) a population of immobilized and/or dead prey mites and/or dead prey mite eggs. In a further embodiment, the prey mite composition according to the invention comprises dead prey mites and dead prey mite eggs. In a more particular embodiment, the prey mite population as used in the invention comprises (a) a population of dead prey mites, and (b) a population of living mites of the species Thyreophagus calusorum. In an even more particular embodiment, the prey mite population as used in the invention comprises (a) a population of dead prey mites from the species Thyreophagus calusorum, and (b) a population of living mites from the species Thyreophagus calusorum.
In a particular embodiment of the invention, the prey mite population comprising mites from the species Thyreophagus calusorum has been treated by a treatment selected from the group consisting of: thermal treatment, such as freezing, heating, cold-shock or heat-shock treatment; chemical treatment, such as gas or fume treatment; radiation treatment, such as UV, microwave, gamma irradiation or X-ray treatment; mechanical treatment, such as vigorous shaking, or stirring, subjecting to shear forces, collision; gas pressure treatment, such as ultrasound treatment, pressure changes, pressure drops; electrical treatment, such as electrocution; immobilizing with an adhesive; immobilization by starvation, such as induced by water or food deprivation; immobilization by suffocation or anoxia treatment, such as by temporarily eliminating oxygen from the atmosphere or replacing oxygen by another gas and any combination thereof. In another particular embodiment, the prey mite population comprising mites from the species Thyreophagus calusorum has been killed, for example by freezing or irradiation. In a further embodiment, the composition of the invention comprises mites from the species Thyreophagus calusorum that have been killed by freezing. It is therefore also an aspect of the invention to provide a method for rearing predatory mites, the method comprising
- providing a prey mite population comprising mites from the species Thyreophagus calusorum,
- killing the prey mite population comprising mites from the species Thyreophagus calusorum,
- contacting the killed prey mite population comprising mites from the species Thyreophagus calusorum with a predatory mite population and a carrier for individuals of the mite populations, and
- allowing individuals of the predatory mite population to feed on said prey mite population.
As mentioned herein before, the carrier can be any solid material, which is suitable to provide a carrier surface to the individuals. In a preferred embodiment, the carrier is a mixture of solid carrier elements. Preferably, the carrier is a substantially homogeneous mixture of solid carrier elements. While the solid carrier elements may have a variety of sizes, a substantially homogeneous mixture refers to the variety of sizes being substantially homogeneously distributed amongst the carrier.
The average longest axis of the solid carrier elements is typically between 1 .0 and 20.0 mm, more in particular between 2.0 and 12.0 mm, preferably between 3.0 and 9.0 mm. In another particular embodiment, the carrier is a mixture of solid carrier elements with a similar size. In a further embodiment, 90% of the solid carrier elements have a longest axis in the range of 0.1 times to 10.0 times the average longest axis of the mixture of solid carrier elements, in particular in the range of 0.2 and 5.0 times, more in particular in the range of 0.5 and 2.0 times. As will be understood from the disclosures herein, two or more different types of carrier elements may be used and mixed. In such case, the size distribution refers to the size distribution per type of carrier elements. Preferably the carrier provides a porous medium, which allows exchanges of metabolic gases and heat produced by the mite populations. Examples of suitable carriers are non-photosynthetic plant material, such as (wheat) bran, buckwheat husks, rice husks, saw dust, tree bark, corn cob grits, etcetera, or inorganic material, such as vermiculite. Advantageously, said carrier comprises grains of a grass species or any part thereof, such as germ or bran. Preferably, said carrier does not comprise living, green plant material, such as leaves or stems of plants. In a particular embodiment, the carrier comprises non-photosynthetic plant material. Particularly preferred is a carrier selected from vermiculite, wheat bran, millet chaff, rice husks and buckwheat husks.
Preferably, said carrier comprises carrier elements with an average longest axis of between 1.0 and 20.0 mm, in particular between 3.0 and 9.0 mm. The carrier material may provide shelter for the prey mites and predatory mites, reducing stress conditions and cannibalism. During mass rearing, the carrier allows for the generation of a three-dimensional rearing matrix and may provide a food source for the prey mites. During storage and distribution for biological control, the carrier acts as a bulking agent and allows for a more homogeneous distribution of the predatory mites in the crops.
Advantageously, the number of individuals of the predatory mite population relative to the number of individuals of the prey mite population is from about 5:1 to 1 :500, such as about 2:1 to 1 :250, and preferably between 1 :1 and 1 :150. In a further particular embodiment, the number of individuals of the predatory mite population relative to the number of individuals of the prey mite population is from about 1 :5 to 1 :30.
High densities of prey mites and predatory mites can be reached thanks to the present invention. In a particular embodiment, the mite compositions of the present invention comprise at least 1000 mites of the prey mite population per gram of the composition, in particular at least 2000 prey mites, more in particular at least 3000 prey mites. In a further embodiment, at least 5000 prey mites, at least 10,000 prey mites, at least 20,000 prey mites, at least 30,000 prey mites. In a preferred embodiment, the mite composition comprises at least 50,000 mites of the prey mite population per gram of the composition.
In another particular embodiment, the mite compositions of the present invention comprise at least 20 mites of the predatory mite population per gram of the composition, in particular at least 30 predatory mites, more in particular at least 50 predatory mites. In a further embodiment, at least 100 predatory mites, at least 150 predatory mites, at least 200 predatory mites, at least 300 predatory mites. In a preferred embodiment, the mite composition comprises at least 500 mites of the predatory mite population per gram of the composition.
Mite numbers are mentioned herein refer to the total number of all mite development stages, thus including eggs, larvae, nymphs, all chrysalis stages and adults, unless the context clearly dictates otherwise. As will be understood from the disclosures herein, the prey mites in the compositions of the invention are not required to be living prey mites.
In a particular embodiment, the mite compositions of the present invention comprise at least 1000 non-egg mites of the prey mite population per gram of the composition, in particular at least 2000 non-egg prey mites, more in particular at least 3000 non-egg prey mites. In a further embodiment, at least 5000 prey non-egg mites, at least 10,000 prey non-egg mites, at least 20,000 prey non-egg mites, at least 30,000 prey non-egg mites. In a preferred embodiment, the mite composition comprises at least 50,000 non-egg mites of the prey mite population per gram of the composition.
In another particular embodiment, the mite compositions of the present invention comprise at least 20 non-egg mites of the predatory mite population per gram of the composition, in particular at least 30 non-egg predatory mites, more in particular at least non- egg 50 predatory mites. In a further embodiment, at least 100 non-egg predatory mites, at least 150 non-egg predatory mites, at least 200 non-egg predatory mites, at least 300 non-egg predatory mites. In a preferred embodiment, the mite composition comprises at least 500 non- egg mites of the predatory mite population per gram of the composition.
In another particular embodiment, the mite compositions of the present invention comprise at least 20 adult mites of the predatory mite population per gram of the composition, in particular at least 30 adult predatory mites, more in particular at least 50 adult predatory mites. In a further embodiment, at least 100 adult predatory mites, at least 150 adult predatory mites, at least 200 adult predatory mites, at least 300 adult predatory mites. In a preferred embodiment, the mite composition comprises at least 500 adult mites of the predatory mite population per gram of the composition. In further embodiment, the mite compositions of the present invention comprise at least 20 living adult mites of the predatory mite population per gram of the composition, in particular at least 30 living adult predatory mites, more in particular at least 50 living adult predatory mites. In a further embodiment, at least 100 living adult predatory mites, at least 150 living adult predatory mites, at least 200 living adult predatory mites, at least 300 living adult predatory mites. In a preferred embodiment, the mite composition comprises at least 500 living adult mites of the predatory mite population per gram of the composition.
Evidently, the above ranges of prey mite and predatory mite numbers can be combined to arrive to particular and preferred embodiments of the invention. It thus follows that the present invention provides, for example, mite compositions comprising at least 1000 mites of the prey mite population and at least 20 mites of the predatory mite population per gram of the composition. As another example, at least 5000 mites of the prey mite population and at least 100 mites of the predatory mite population. As another example, at least 50,000 mites of the prey mite population and at least 500 mites of the predatory mite population.
In a preferred embodiment, said predatory mites are mesostigmatid or prostigmatid mite species. In a further embodiment, said predatory mites are selected from:
- Mesostigmatid predatory mite species such as: i) Phytoseiidae such as from: a)- the subfamily of the Amblyseiinae, such as from the genus Amblyseius, e.g. Amblyseius andersoni, Amblyseius aerialis, Amblyseius swirskii, Amblyseius herbicolus or Amblyseius largoensis, from the genus Euseius e.g. Euseius finlandicus, Euseius h ibisci, Euseius ovalis, Euseius victoriensis, Euseius stipulatus, Euseius scutalis, Euseius tularensis, Euseius addoensis, Euseius concordis, Euseius ho or Euseius citri, from the genus Neoseiulus e.g. Neosei ulus barkeri, Neoseiulus califomicus, Neoseiulus cucumeris, Neoseiulus longispinosus, Neoseiulus womersleyi, Neoseiulus idaeus, Neoseiulus anonymus, Neoseiulus paspalivorus, Neoseiulus reductus or Neoseiulus fallacis, from the genus Amblydromalus e.g. Amblydromalus limonicus from the genus Typhlodromalus e.g. Typhlodromalus aripo, Typhlodromalus laila or Typhlodromalus peregrinus, from the genus Transeius e.g. Transeius montdorensis, from the genus Phytoseiulus, e.g. Phytoseiulus persimilis, Phytoseiulus macropilis, Phytoseiulus longipes, Phytoseiulus fragariae; b) the subfamily of the Typhlodrominae, such as from the genus Galendromus e.g. Galendromus occidentalis from the genus Typhlodromus e.g. Typhlodromus pyri, Typhlodromus doreenae, Typhlodromus rhenanus or Typhlodromus athiasae; c) the subfamily of the Phytoseiinae, such as from the genus Phytoseius e.g. Phytoseius macropilis, Phytososeius finitimus or Phytoseius plumifer. ii) Ascidae such as from the genus Proctolaelaps, such as Proctolaelaps pygmaeus (Muller); from the genus Blattisocius e.g. Blattisocius tarsalis (Berlese), Blattisocius keegani (Fox); from the genus Lasioseius e.g. Lasioseius fimetorum Karg, Lasioseius floridensis Berlese, Lasioseius bispinosus Evans, Lasioseius dentatus Fox, Lasioseius scapulatus (Kenett), Lasioseius athiasae Nawar & Nasr; from the genus Arctoseius e.g. Arctoseius semiscissus (Berlese); from the genus Protogamasellus e.g. Protogamasellus dioscorus Manson; iii) Laelapidae such as from the genus Stratiolaelaps e.g. Stratiolaelaps scimitus (Womersley) (also placed in the genus Hypoaspis) Gaeolaelaps e.g. Gaeolaelaps aculeifer (Canestrini) (also placed in the genus Hypoaspis); Androlaelaps e.g. Androlaelaps casalis (Berlese); iv) Macrochelidae such as from the genus Macrocheles e.g. Macrocheles robustulus (Berlese), Macrocheles muscaedomesticae (Scopoli), Macrocheles matrius (Hull); v) Parasitidae such as from the genus Pergamasus; e.g. Pergamasus quisquiliarum Canestrini; Parasitus e.g. Parasitusfimetorum (Berlese), Parasitus bituberosus Karg; Parasitellus e.g. Parasitellus fucorum (De Geer); vi) Digamasellidae such as from the genus Digamasellus; e.g. Digamasellus quadrisetus or Digamasellus punctum; from the genus Dendrolaelaps; e.g. Dendrolaelaps neodisetus; and
-prostigmatid mite species such as from: i) Tydeidae such as from the genus Homeopronematus e.g. Homeopronematus anconai (Baker); from the genus Tydeus e.g. Tydeus Iambi (Baker), Tydeus caudatus (Duges); from the genus Pronematus e.g. Pronematus ubiquitus (McGregor); ii) Cheyletidae such as from the genus Cheyletus e.g. Cheyletus eruditus (Schrank Cheyletus malaccensis Oudemans; iii) Cunaxidae such as from the genus Coleoscirus e.g. Coleoscirus simplex (Ewing), from the genus Cunaxa e.g. Cunaxa setirostris (Rermann); iv) Erythraeidae such as from the genus Balaustium e.g. Balaustium putmani Smiley, Balaustium medicagoense Meyer & Ryke , Balaustium murorum (Hermann); v) Stigmaeidae such as from the genus Agistemus e.g. Agistemus exsertus Gonzalez; such as from the genus Zetzellia e.g. Zetzellia mali (Ewing); and vi) Tarsonemidae such as from the genus Acaronemus, e.g. Acaronemus destructor (Smiley and Landwehr); such as from the genus Dendroptus near suski Sharonov and Livshits; such as Lupotarsonemus floridanus Attiah.
In a preferred embodiment, the predatory mites belong to the Phytoseiidae. In a particular embodiment, the predatory mites are selected from Amblyseiinae, Transeius, Neoseiulus and Amblydromalus; more in particular the predatory mites are selected from the group consisting of Amblyseius swirskii, Transeius montdorensis, Neoseiulus californicus, Amblydromalus limonicus, Neoseiulus cucumeris, Amblyseius andersoni, Iphiseius degenerans and Stratiolaelaps scimitus; more in particular from the group consisting of Amblyseius swirskii, Transeius montdorensis, Neoseiulus californicus, and Amblydromalus limonicus. In a preferred embodiment, the predatory mites are selected from Amblyseius swirskii and Neoseiulus californicus, preferably Neoseiulus californicus.
The present invention also provides a commercial packaging for storing and distributing the composition of the invention. Therefore, the present invention further pertains to a container comprising a composition according to the invention. In particular, the container has an internal volume of between 0.004 I and 5 I, preferably between 0.5 I and 2 I. According to a preferred embodiment the container preferably comprising an exit for at least one motile life stage of the mite, more preferably an exit suitable for providing a sustained release of said at least one motile life stage. In a particular embodiment, the container (e.g. a bottle or a sachet) has at least one exit with a removable seal. In particular, seal as used herein refers to a closure that is glued or heat-sealed. Preferably, the container is adapted to attach it to a crop, for example by comprising a hook to hang it from a crop leaf or branch or by comprising a sticky surface to stick it to a surface of the crop or with a stick attached to the sachet.
The present invention further pertains to a method for rearing predatory mites, the method comprising
- providing a composition of the invention, and
- allowing individuals of the predatory mite population to feed on said prey mite population.
Preferably, the individuals of the predatory mite population are allowed to feed on said prey mite population while maintaining said mite composition at 5 to 35 °C and 30 to 100% relative humidity; in particular at 15 to 35°C and 50 to 100% relative humidity. The present invention further pertains to a method for controlling a pest in a crop, the method comprising providing to said crop a mite composition according to the invention. The mite composition of the invention may be distributed directly onto the crop. Alternatively, the mite composition of the invention is provided in the proximity of the crop. For example, the mite composition of the invention may be provided to a crop by placing a container holding the mite composition in the vicinity of the crop and allowing the predatory mites to exit from said container. This way, predatory mites spread themselves through the crops and control the crop pests. In a preferred embodiment, the pest is an arthropod pest.
The present invention further provides the use of mites from the species Thyreophagus calusorum for feeding predatory mites. Conveniently, it has been found that these mites can be reared efficiently and provide an easy food source for predatory mites. The present application allows to distribute mites from the species Thyreophagus calusorum in a crop such that predatory mites that are present in the crop can feed thereon. The mites from the species Thyreophagus calusorum (herein also referred to as prey mites or prey mite population) may be distributed on their own or may be distributed in combination with predatory mites. When distributed in combination with predatory mites, the predatory mites and prey mites may be distributed at the same time or at distinct times.
These and other embodiments of the invention are indicated in the appended claims.
The invention will now be further described with reference to the following examples, which show non-limiting embodiments of different aspects of the invention.
Examples
Example 1 : Reproduction of Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites
This experiment was conducted to assess the reproduction of Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female N. californicus were individually transferred from the mass-rearing to a PVC arena (1 x 2 cm) of which the edges were covered with tissue paper. A small volume of prey mites (0.1 mL) was provided as food for the females. The trial was conducted in a climatic chamber set at 23 ± 1 °C, 90 ± 5 % and a 16:8 h (L:D) photoperiod. Results were analyzed by counting and removing the deposited eggs on a daily basis. Data of the first day was omitted to reduce the effects of pre-experimental conditions (van Houten et al. 1995). The number of eggs and mobiles was compared using a generalized linear model with a Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014). As demonstrated in Fig. 1 , a significantly higher number of eggs were deposited when N. californicus was fed with Th. calusorum compared to Th. entomophagus (GLM: p<0.05). It can hence be concluded that Th. calusorum is more efficacious prey mite for rearing predatory mites.
Example 2: Growth comparison of predatory mite Amblyseius swirskii on Th. entomophagus and Th. calusorum prey mites
This experiment was conducted to compare the population growth of the phytoseiid predatory mite Amblyseius swirskii on two species of astigmatid prey mites, namely Thyreophagus entomophagus and Thyreophagus calusorum. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female A. swirskii were transferred to an experimental breeding dish (5 x 1.5 cm) (SPL Life Sciences, Korea), which was filled with sawdust. A small volume of prey mites (0.5 mL) was added to the mix as food for the predatory mites. The trial was conducted in a climatic chamber set at 23 ± 1 °C, 90 ± 5 % and a 16:8 h (L:D) photoperiod. Results were analyzed by comparing the number of predatory mite mobiles and eggs after 10 days using a generalized linear model with a quasi Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014).
As shown in Fig. 2, the number of eggs and mobiles of A. swirskii fed on Th. entomophagus were highly similar to those fed on Th. calusorum (GLM: p>0.05 for both eggs and mobiles). This signifies the suitability of using Th. calusorum as prey mite for rearing predatory mites.
Example 3: Growth comparison of predatory mite Neoseiulus californicus on Th. entomophagus and Th. calusorum prey mites
This experiment was conducted to compare the population growth of the phytoseiid predatory mite Neoseiulus californicus on two species of astigmatid prey mites, namely Th. entomophagus and Th. calusorum. Both prey mites were produced in rearing containers on a diet consisting of wheat bran and baker’s yeast. Ten young gravid female N. californicus were transferred to an experimental breeding dish (5 x 1 .5 cm) (SPL Life Sciences, Korea), which was filled with sawdust. A small volume of prey mites (0.5 mL) was added to the mix as food for the predatory mites. The trial was conducted in a climatic chamber set at 23 ± 1 °C, 90 ± 5 % and a 16:8 h (L:D) photoperiod. Results were analyzed by comparing the number of predatory mite mobiles and eggs after 10 days using a generalized linear model with a quasi Poisson distribution using the statistical software R (version 3.6.1 ) (R Core Team 2014).
As can be seen in Fig. 3, the number of mobiles of N. californicus was significantly higher when fed on Th. calusorum compared with Th. entomophagus (GLM: p<0.05), while the number of eggs was similar for both pre mites (GLM: p>0.05). Henceforth, it can be concluded that use of Th. calusorum will result in a higher amount of predatory mite mobiles, leading to clear economic benefits over the use of Th. entomophagus.
Example 4: Comparison of egg/mobile ratio between C. transversostriata and Th. calusorum prey mites
For feeding phytoseiid predatory mites, a prey mite should preferably have a high proportion of eggs. The capture success-rate is higher in egg stages as compared to nymphal or adults stages, also referred to as mobiles (Midthassel et al. 2013). This experiment was conducted to compare the proportion of eggs to mobiles between two species of astigmatid prey mites, namely Czenspinskia transversostriata and Th. calusorum, the prey mite of the invention. Random production units containing 1 liter of volume were selected from the massproduction of both Th. calusorum and C. transversostriata. A subsample was taken and the number of egg and mobiles stages were counted separately under a stereo microscope. The egg/mobile ratio of both species was compared with a generalized linear model using the statistical software R (version 3.6.1 ) (R Core Team 2014).
As can be seen in Fig. 4, the egg/mobile ratio of Th. calusorum was about 4 times higher than that of C. transversostriata (GLM: x2= 4.0; df=1 ; p<0.001). Thus, it can be concluded that the Th. calusorum prey mites are a more efficient food source for phytoseiid predatory mites, compared with C. transversostriata.

Claims

Claims
1 . A mite composition comprising:
- a predatory mite population,
- a prey mite population, and
- a carrier for individuals of said populations, wherein said prey mite population comprises mites from the species Thyreophagus calusorum.
2. The composition according to claim 1 , further comprising a food source for said mite populations.
3. The composition according to claim 1 or 2, wherein the food source for said prey mite population comprises wheat germ; pollen, or a fungus, such as yeast.
4. The composition of any one of the previous claims, wherein said carrier comprises grains of a grass species or any part thereof, such as germ or bran.
5. The composition of any one of the previous claims, wherein said carrier comprises carrier elements with an average longest axis of between 1 .0 and 20.0 mm.
6. The composition of any one of previous claims, wherein the number of individuals of the predatory mite species relative to the number of individuals of the prey mite is from about 5:1 to about 1 :500; in particular from about 2:1 to about 1 :200; more in particular between 1 :1 and 1 :50.
7. The composition of any one of previous claims, wherein said predatory mite species are mesostigmatid or prostigmatid mite species, in particular Phytoseiidae.
8. The composition of any one of the previous claims, wherein the composition comprises at least 200 prey mites of the species Thyreophagus calusorum per gram of the composition and at least 10 predatory mites per gram of the composition.
9. A container comprising the composition of any one of the previous claims.
10. A method for rearing predatory mites, the method comprising
- providing the composition of any one of claims 1 to 8, and
- allowing individuals of the predatory mite population to feed on said prey mite population. The method of claim 10, further comprising maintaining said mite composition at 5 to 35 °C and 30 to 100% relative humidity. The method of claim 11 , further comprising packaging the composition in a container with an exit that is designed to be opened to release at least individuals from the predatory mite population from the container. Use of mites from the species Thyreophagus calusorum for feeding predatory mites. A method for controlling a pest in a crop, the method comprising providing to said crop a mite composition according to any one of claims 1 to 8. The method of claim 14, comprising placing the container of claim 9 in or at close proximity to the crop.
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