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(11) Pateot Number: KE 409

(21)    Application Number: KE/P/2009/ 000862

(22)    Filing Date: 26/07/2007

(30)    Priority data: 0615358.9  02/08/2006  GB

(86)    PCT data PCT/GB07/002858    26/07/2007 WO  2008/015393  A3 07/02/2008
 
(73)0wner: SYNGENTA BIOLINELIMITED of TelstarNursery, Holland Road, Little Clacton, Clacton-on-Sea, Essex C016 9QG., United Kingdom

(72)    Inventor:  FIDGET, Melvyn John of Syngenta Bioline Limited, Telstar Nursery, Holland Road, Little Clacton, Clacton-on-Sea, Essex COI6 United Kingdom and STINSON, Clive Stewwt Alexander of  SyngenlB Bioline Limited, Telstar Nursery, Holland Road, Little Clacton, Clacton-on-Sea, Essex C016 United Kingdom

(74)    Agent/address ror correspondence:  MURIU MUNGAI &CO. ADVOCATES, P. 0. BOX 75362-00200, NAIROBI, KENYA
 
(54)    Title: METHOD FOR REARING PREDATORY MITES

(57)Abstrad:The present invention relates to the field of biological control. Specifically, it relates to the use of predator mites as biological control agents for reducing damage to crops by insect pests. In particular, it relates to a new method for rearing predator mites, and a method for controlling pests in a crop using predator mites reared using said method
 
METHOD FORREARJNG PREDATORY MITES

The present invention relates to the field ofbiological control. Specifically, it relates

to the use of predator mites as biological control agents for reducing damage to crops

by insect pests. In particular, it relates to a new method for rearing predator mites,

and a method :fur controlling pests in a crop using predator mites reared using said method.

Plant pests are a major factor in the loss of the world's important agricultural crops. About $8 billion is lost every year in the U.S. due to infestations of plants by non-ma;mmalian pests including insects. In addition to losses in field crops, insect pests are also a burdeo to vegetable and fntit growers, to producers of ornamental flowers,

and to home gardeners. For example, pest thrips such as Franklinieila occidentalis,

and whitefly such as Bemisia tabaci cause extensive damage to horticultural crops

such as salad vegetables, cut flowers and omameotal plants, resulting in significant

economic loss to the growers.

Insect pests are mainly controlled by intensive applications of chemical pesticides,

which are active through inhibition of insect growth, prevention of insect feeding or

reproduction, or cause death. Although the use of such chemicals can result in good

control of insect pests, the widespread use of chemical pesticides can result :in the appearance of resistant inseot varieties. Further, high levels of chemicals on horticultural crops, in particular salad vegetables, are undesirable to many consumers.

Therefore, an alternative method of pest control involving the use of beneficial insects or mites has been developed. The beneficial insects or mites are predatory towards
pest insects such as thrips, and can be applied to crops tc control insect pests.

Beneficial insects or inites can be provided to crop plants in a variety of ways, for

example manually or through a controlled release device. The use of beneficial

insects or mites fonns part of integrated crop management and integrated pest

management programs, combining cultural, biological and chemical mea11s to achieve

sustainable pest control.
 One example of a beneficial .insect or mite system is the use of Amblyseius cuc10neri8

as a predatory mite fur the control of pest thrips.  Existing systems involve the mite

being provided either loose in bran and vermiculite for sprinkling onto crops, or in sachets for longer periods ofprotection. Amblyseius r::ucumeris feed on .first instax thrips larvae, and control relies on complete cover of a. crop with the predator before thrips establish. Recently, Amblyseius swirsldi has been introduced as an alternative

predatory mite to Amblyseius c:uc:umri. It is rapidly becoming the pred!Uory mite of choice amongst growers beoall8e it con be ll8ed for the control ofboth thrips and whitefly.

Mass rearing systems that were suitable for generating large numbers ofheneficial

insects and mites were first introduced approximately 70 years ago. Original mass rearing systems were based on the provision of a natural food source of the insect or mite to be ~ared. For example, mass reming systems can be based on the provision ofhost species SllCh as Tetranychus wticae for Amblyseius swirskii or Phytoseiulus

persimilis, pollen grains for othor predatory mites, or aphids for Aphidoletes

a:phidimyza.  More recently, maoo rearing systems for predatory mites have been

successfully developed using fuctitious hosts.  The most oommonly used hosts are

stored product mites such as Tyrophagus putrescentiae, Acarus siro, and

Carpoglyphw lactis.  Through the provision of predator mites, pl~Y mites, and a food

source for the prey mites, a stable breeding colony CBl1 be established to allow the

rearing of a continuous supply ofpredator mites.

GB2393890 discloses that A. cuc:unzeriJ can be reared using Tyrophagw; putrescentiae,

T. tropiCWI,_ or Acarus siro.  Rmnakers eta/. (Bulletin-SROP 6(3), 203-206) discloses

that rearing of AmblyseiUs mckenziei and A. cucu.meris can be carried out using wheat

bran as the primary food source and A= farris as substilllte prey. Nomikou et a/.

(Experimental and Applied Acarology 27(1-2), 57-68) discloses that Tyrophagw;

putrescentiae can be used as a. food sour~e for Typhlodromips (AmblyesiJJs)swirsldi,

fur obtaining high predator/prey ratios.  Steiner eta/. (Australian Journal of

Entomology 42, 124-130) discloses that 1j>rophagw; putresceTiiiae can be used as a

food source for Typhlodromips mon.tdorensis.  W02006/051552 discloses that

Amb/y.seius s-wirsfdi can be reared on Alrtigmatid mites, such as mites from the
 
families Carpoglyphidae, Pyroglyphidae, G/yr:iphagidae or Acaridae. In particulBr, it cliscloses that CarpoglJPhus lacru can be used as a prey mite for rearing A. swirskii.
CN1440646 describes the use of various species of stored product mite for raising the predatory mite, Amblyseiu.r cru:umeri.r.

One disadvantage to the existing mass rearing systems is that the speed of growth of

the predator colony is limited. TherefOre, there exists a need for an improved mass

rearing system that will reault in a faster breeding rate of the predator colony.  The

present ~vention overcomes this problem through the provision of a better food

source. In conventional mass rearing systems, predator mites tend to feed mWnly on

the eggs ofprey mite.  One reason for this is that the juvenile and adult forms ofmost

grain mites that .::u-e used as prey are quite haity.  The pres~t invention relates to the

use of a prey.mite that is less hairy than most other Astigmatid mites. and therefore predators feed well on adulbl and juveniles as well as eggs.

Another problem associated with existing mass rearing systems is that breeding populations of predator mites are very sensitive, and can be easily affected by changes in envlromnental conditions. Many mites that are conventiona.Uy used as prey mites are very mobile. Their rapid and continuous movement disturbs predator mites,

which in tum causes stress, and upsets reproduction, egg laying and feeding. In tum, the quality and number of predator mites reared decreases. The present invention
solves this problem through us'e of a less active mite as thfl prey mite,. which causes

less .disturbance to predator colonies, in turn resulting in a greater Illl.Dlber of healthier

predator mites being produced.  Another problOlll with many mites that are

conventionally used as prey mites is that they can generate high levels of metabolic

heat and carbon dioxide, which can also have a detrimental effect on the health and

size of the predator colony. The present invention solves thiB problem through use of

less active mites that generate less metabolic heat and carbon dioxide.

According to the present invention there is provided a method for rearing predatory

mites, comprising providing Thyreophagus entarnophagus as prey mites, and allowing

the predatory mites to feed on said prey mites.
 

In the present context, the term "rearing" refers broadly to breeding, reproducing, surviving and growing ~f predatory mites. Rearing can be in an open or closed environment. Typically it will take place in a closed environment, such as a growth room or inoubator.

The growth of a predator colony may be assessed by monitoring the time between gencrntion periods, the rate of egg production (oviposition rates), and/or fecundity.

The tem1 ''prey mites" refers to mites that are present specifically as prey for the predatory mites to feed on. The predatory mites may food on any life stage of the prey mite, for example eggs, juveniles or adults.

The term ''predatory mites" refers to mites that are predators of any pest.  In particular,

it refers to mites that are predators of crop pests. They may be predators of pests such as insects, nematodes or arachnids. Typically, the predator mites "Will be useful for conb:ol of common crop pests such as thrips and whitefly. Most predatory mites
beloug to the family Ph;1oseiidae (order Acarina). The most common predatory

mites used for biological control in glasshouses are Amblyseiu;; swirskti, Phytoseiulus

per.s~, Amblys~us californicus, Amblyseiw cucumeris, Amblyseiw degeneraru

and Hypoaspi.s miles.

Amb/yseii!S (Jjphlodromips) swirskii is used principally for the control of thripB, such

as Fra:nkl.iniella. occidenta.lis or Thrips tabaci, and whitefly, such as Trialewodes

vapora:riorum or Bemisia. tabaci.  It is found :in eastern Mediterranean regions

including Israel, Italy, Cyp!Us and Egypt. It is well suited to a warm and hwnid

cHmate.

PhytoseiulrtS persimilis is used in biological control programs for two-spotted spider

mites (Tetranychu;; urticae), and related Tetranychus species.  The mites Bie predators

as nymphs and adults, mostly feeding on spider mite eggs and nymphs, but also

consuming ad:ults.  Since P. persirn:iliB is an obligate spider mite predator and ca.nnot

survive on alternate food sources such as pollen, survival tends to be poor ifprey is in

short supply.
 
AmblyseiWI (Neosetulus) californicus is an active predatory mite that has specialised

in feeding on mites in the family Tetranychidae. T1ris ine!udes the common Red

Spider Mite or Two-Spot Mite TetranycizuJ urticae. and the Carmine Mite

Tetranychus cinnabarinus. It is less specialised than Phytoseiulu.s persimilis, and while it prefers to feed on spidor mites, it can feed and reproduce on other arthropod prey or pollen. Where few spider mites w.•e present, it is able to survive by feeWng on these alterriative sources of food, and so can persist in a crop longer than Phytoseiulus
persimilis.

Amblyseius (Neoseiulus) cuctonerls iB used to control thrips, including the Western Flower Thrips (Franldiniella occidentalis). The main food somce is fust-instar thrips lmvae. Later stage larvae and adults are less susceptible due to the small size of the mite. Since A. cucumeris can also feed on other mites (such as cyclamen mites and broad mites) as well as pollen, it can survive in many different situations.

Amblyseius (lphiseius) degenerans, is more aggressive than A. cucumeris in attacking thrips, and usually colonises flowers in greater numbers. A. dege11ertz113 will also feed on spider mites and pollen.

Hypoaspis miles mites, from the family Laelapidae,live in soil and other growing media. They feed on fungus gnat larvae, springtails and also thripB pre-pupae and pupae. With the range of food eaten by these predators, they can become established and parsist for long periods. Since this predator lives under the surface of the growing media, it can escape contact with many pesticides that are used to control pests that feed on leaves and flowers.

Other predatory mites include Euseius tularensis1 'I)phlodromus occidentalis1

Typhlodromus pJiri, Zetzellia mali, Amblyseius (Iphiseius) degen.erans, Amblyseius (Kampimodromus) aberrans, Amblyseius (Neoseiulus) ba:rkeri, Amblyseius andersoni, blhlyseius (Neoseiulus) fallacis, Amblyseius (Eu.seius) finlandicus, M1blyseius (Iyphlodromalus) lailae, Amblyseius (IYphlodromalus) limon.icus, Amblyseius (TYphlodromips) montdorensis, Amblyseius (Euseius) ovalis, Amblyseius (Euseiu.s)
 
scutalts, AmhlyseitJS (Euseius) stipulatus and Amblyseiu.r (Neoseiulus) wamersleyi

(also known asAmblyseius longispinosus). This list is not exhaustive. McMurtry et a!. (Annual Review of Entomology 42, 291-321) categorises the diversity oflife-stages in the Phytoseiidae based mafu!y on food habitats and related biological and
moiphological traits. As mite taxonomy is continually changini, the number and

species of mites classified as• predatory mites may change (see for example the

Catalogue orPhytoseiidae by Moraes et al. published in 2004 that lists 2,250 species,

compared to the prav:ious edition published in 1986 that listed only 1500 species).

The present invention relates to the use of Thyreophagus entomophagus as a food

source for any predatory mite. In particular, the predatory mites may be selected from

the group consisting ofAmblyseius (J)phlodromips) swirskii, Phytoseiulus persimilis, Amhlyseius cucumeris, Amblyseius degenerans, Hypoaspis miles, Euseius tularemsi.s, Typhlodromus occidental is, Typhlodromus pyri, Amblysieus fallacies, Zetzellia mali,
Amblyseius (Jphi.reius) degeneran.s, AmblyseiWJ (Kampimodromw) aberrans,

Amb/yseius (Neoseiulus) barkeri, Amblyseius  andersoni, Amblyseius

(Neoseiulus) fallacis, Ambiyseius (Euseiw) finlaru:licus, Amblyseius (Typhiodroma/us)

lai/ae, Ambiyseius (Typhlodrornaius) /irnonicus,Amblyseiwi (Typhiodromips)

montdorensi.r, Amhlyseius (Euseius) ovalis, Amblyseius (Euseius) scutalis, Amblyseiw

(Eweius) stipulatus andAmbryseius (Neoseiulus) womersleyi (also known as

Ambiyseius iongispinosus).

The prey mite Thyreophagw entomophagw (Laboulbane, 1852) is also known as

Acarus entomophagu.s, Histiogaster entomopho.gm, Histiogaster aleurophagw,

Tyroglyphw malw, and Dermaleich~~:~ malus.

The body of the Thyreophagus entomophagw male is elongated and oval, with a

colourless shirring cuticle and pale brown stumpy legs. At its posterior end, the

hysterosoma is extended backwards as a horizontal semicircular shelf in the same

plane as the ventral surface of the body, and with a well~sclerotized ventral surface.

The propodsomal dorsalsbield extends as far back as the scapular setae, but the rest

of the surface of the idiosoma has a naked appearance because of the scarcity of setae.

The body of the Thyreophagus entomophagw female is longer and more slender than

tilat of the male, the posterior end being slightly pointed and not prolonged into an

opisthiosomallobe. The genital opening lies between coxae ill and IGV, and is well separated from the anus, which extends as far back as the posterior edge of the body; two pairs of long anal setae arise on either side of it The chaetoabcy of body and legs is the same as that of the male, except that a small spine is present at the base of all the claws.

Thyreophagu.s entomoplw.gus is a largely sedent!D."y mite which shows no inclination to move a.wa.y from suitable food supplies, even whtm disturbed. It does not avoid pred<rtors, and has no visible defence mechanisms against them. Predators can be seen recoiling from adults of Tyrophagus putrescenliae, suggesting the existence of an active, possibly chemically-based, defence mechanism.

Usually, pre:&ror mites feed on the egg stages of mite hosts.  One reason for this is

that the juvenile and adult stages of the many mite bosts such as Carpoglyphus lacti:J aud Tyrophagus putrescentiae are quite hairy. This makes the prey mites less
attractive to pred.rors. However, sim;e the adult fonn of J1zyreophagus

entornophagu.s is less hairy, predator mites readily feed on the juveniles and adults, as well as eggs. Therefore, Thyreophagw entomophagus is a better prey because predator mites can attack more stages of the mite life cycle.

Furtl:er, Thyreophagus mtomophagu.s is a fairly sessile mite. It is less active and slower moving than other mites that have previously been used as prey, such as
Carpoglyphus /aclis, Acarus siro lllld 'I'yrophagus putnscentiae. Breeding cultures of

mites are very sensitive, and can be easily disnJ.pted.  In breeding cultures, predators

are easily disturbed by very active or mobile prey mites.  It is important to minimise

dismption to the predator mites, because disturbance will result in decreased egg-l•ying. Also, disturbing the juvenile predators will upset their feeding and cauae stress, in turn reducing their chances of developing into healthy adult predators. With mobile prey species, as the ratio of prey to predator increases (for example due to reduced egg laying, or reduced numbers ofjuveniles developing into adults) there will be a further negative effect on the reproduction ofthe predator mites due to a greater
 
leveJ of disturbance in the rearing medium. Another effect of the ratio of prey to predator increasing is that there is an increase iD. metabolic heat, fi."BSS and fungal organisi:ns within the breeding culture. This will again have a. detrimental effect on
the reproduction and quality of lhe predators in the culture.

Mites such as !(arpoglyphus lac:ti3 and Tyrophagus putrescentiae thaJ are

convontionally used as prey for rearing predatory mites, produce alarm pheromones

when attaclced, or when present inlrigb numbers. This contributes to their activity,

and further increases the level of disturbance in the rearing medium. No similar

pheromone is repcrted for Thyreophagus entomophagus.

In one aspect of the present invention, the prey mites will be present as a breeding

population. In this way, there will be an adequate food source for the predatory mite•

that is continuously replenished. Predatory mites may feed on one or more of the life cycle sta.ges of the prey mite incluCling eggs, lanral stages, nymphs, juveniles and/or
adults.

In one aspect of the present invention, the predatory mites are from the family Phytosciidae. TI1e taxonomy of mites is still evolving. The Zoo taxa catalogue (De
Moraes, McMurty, Denmark & Campos, 2004, Zootaxa 434, 1-494) describes the'

current understanding of the taxonomy of the Phytoseiiilae, snd lists all known

members of this .fumily. In wether aspect of the invention. the predatory mites are

:from the subfamily Amblysciinae. In a further aspect of the invention, the pi""'...datory mites are from a genus selected from the group consisting afAmblyseius,

Typhlodromips, Neoseiulus, ljphlodramalus, Euseius, Typhlodromus, Iphiseius and

Kampimodromu.s.

In a further aspect of the invention, the predatory mites are from the genus Amblyseius.

Examples ofpredatory mites from this genus include Amblysetu.. (Kampimodromus)

aberrans, Arnblyaeiu.s (Neoseiulus) barkeri, Amblyseius andersoni, Amblysei.us

(Neoseiulus) californicus, A:mblyseius (Neoseiulu.s) cucumeris, Amblysei.us

degenerans, Amblyseius (Neoseiulus) fallacis, Amblyseius (Euseius) jinlandicus,

Amho/seiu.s (Typhlodromalus) lailae, Amblyseius (TYphlodrornalus) limonicus,

Amblyseius (Typhlodromips) montdorensis, Amblyseiu;; (Euseius) ovalis, Amblyseius (Euseius) scutalis, Amblyseius (Euseius) stipulatus, Amblyseius (Typhlodromips) swirsldi, and Amblyseius (Neoseiulus) womersleyi (also known as Amblyseius longispillosus). The predatory mites may, for example, beAmblyseius andersoni, Amblyseius califomicus, AmblyseiU3 cucumeri.s, Amhlyseius fallacis, Amblyseius limonicus, Amblyseius montdore.nsi.J, Amblyseius ova.lis, Amblysetus stipulatus, Amblyseius swirsldi or Amblyseius womersleyi. In a preferred aspect of the lnvention, the predatory mites are Amblyseius S'rl'irsldi. This mite is also known as
Typhlodromips swirskii, and these names are used synonymously herein.

The present lnvention may be used to provide a source of predator mites for the control of pest insects. A particular predator will be selected on the basis of the target pest to be controlled, and the crop to which the predator will be applied.

For example, the predator mites maybe used to control one or more of the pests from the followln.& non~exhaustive list: Tetranychus spp. including but not restricted to
Tetranychus urticae, T. cinnabarinus, T. kanza-wai, T. turkestan.i, T. occiden:talis; Oligonychus spp.; Pano11Jichus ulmi andP. citri; Eriophyid mites inclnding but not restricted to Aculop.s lycopersici, Aculus schlectendali, Phyllocopt1"'/J.a oletvora, A ceria ficus, Rhyncaphytopi'!J.> ficifoliae; Tarsonemid mites inclnding but not restricted to Polyphagotarsonem:WJ latus, Phytonemus pallidus; thrips pests including but not restricted to Frankliniella occidental~, F. intorua, F. sclwltzei, Thrips tabaci,

T. palmi, Echinothrips americanus, Heliothrips haemorrhoidali.s;  and whitefly pests

including but not restricted to Tn'aleu.rodes vaporarion.1m, Bemisia tabaci, Aleyrodes

proletella, A. lonicerae.

It will be preferable to choose a predator thet will not cause damage to the crop plant itself. The predator mites may be used to control pests on one or more of the crop plants in the following, non-exhaustive list: tomato, sweet pepper, chilli pepper, aubergine, cucumber, beans, squash, melon, strawberry, raspberry, bm1ana, papaya, apple, pear, plum, grape, rose, chrysanthemum, Gerbera, Begonia, Cyclamen,

Poinsettia, Citrus, Slrimmia, Choiysia, Daphne and Magnolia.  Suitably, the crop is

selected from the group consisting of peppers, cucumbfii'B, aubergines, roses, gerberas, melons and beans, especially when

In one embodiment, the present invention may be used to provide a source of predator mites for the control of thrips (such as Franldiniella occidentalis or Thrips tabaci) and
I or whitefly (such as Trialeurodea vaporariorum or Bemisia tabaci). In particular, the present invention may be used to provide a source of Amblyseius s1virsldi mites for
the control ofthrips and/or whitefly.

It is important to use an appropriate starting ratio of predator mites to prey mites. For

Amblyseius cucumeris reared on TYrophagus putrescentiae, predator.prey ratios are

noiJD.Blly botw""" 1:4 and I: I 0. Higher ratios usually mean that the colony will be overwhehned by Tyrophagus. For Amblyseius swirskii reared on Carpoglyphv.s lactis, the ratio is normally between 1:4 and 1:10. In conlrast, for predator mites reared on

Thyreophagus entampltagus, a higher starting ratio of predator:prey is required.  The

optimum ratio will vary depending on the nature of the predator species being reared.

For predators mites reared on Thyreophagus entomophagus, the preferred starting

predator:preyratio is between 1:10 and 1:100. Suitably it is botween l:ZO aod 1:80. More suitably it is between 1:30 and 1:70. In one embodiment, the startiogratio is a1 least 1:30. In another embodiment, it is b-een 1:40 and 1:60. In a further embodiment, the starting predator:prey ratio is approxn;,.tely I :50. In a further embodiment still, the starting ratio is at leBBt 1:50. Since the cultures ofboth the

predator and prey mites are actively reproducing, the Iatio of predator to prey mites may change over time.

For a reliable, consistent mass rearing ofAmblyseius swirsk:i.i, a suitable starting ratio of Thyreophagw entomophagus : Amblyseiw swinki.i is approximately 1:50. Below
this the production may be unstable as there is a risk oithe Thyreophagus

entomophagus mites being completely consumed before the breeding cycle b .. been

completed. The consequence of this will be a reduced end yield due to cannibalism

and reduced breeding, and less healthy predators due to starving.

For mass-rearing of Amblyseius a.ruiersoni on Thyreoplzagus entomophagus, all the prey mites will be completely consumed if the predator:prey ratio at the initial set up inoculBtion is below approximately 1:30. More vomcious and/or faster reproducing predator species such as Ambly.seius cucum.eri.s would require an even higher initial predalor:prey ratio in order for the predator:prey balance to be Dlllinlained through the entire cycle and so provide the optimum end cycle predator concentration in the production system. The person skilled in the art is familiar with the need to optimise the starting ratio of predator mites to prey mites for each species combination.

Thyreophagus entomophagus can be reared on a variety of dietB. Suitably, it is reared on a high carbohydrate yeast-based diet. In this coiJ±ext, a diet that contains at least 5% sugar is .a high carbohydrate diet. The diet is mixed with bran as a carrier ma ratio of approximately 20% diet:80% bran (v/v). The mites can be reared in the medium in aplastic container, typically ranging in size from 30m1 to 10 li1res. The containers are vented with 60micron nylon mesh discs.

During rearing of Thyreoplw.gus entomophagus there are a Dllm.ber of production stages. At each stage, small numbers of Thyreophagus entomophagus mites are inoculated into the containers and placed in a 10om having suitable conditions for allowing the mites to reproduce. Suitably the temperature in the room is in the range from l5°-30°C and the humidity between ?o-95% r.h .. Most suitably, the tempemture is approximately 28°C. The cycle time from inoculation to end product ranges from 7-21 days. Suitably, it is approximately 14 days. The rate of reproduction in these conditions will be sufficient to produce an end culture concentration of around 2 million mobile stages of Thyreophagus entomophagus and eggs from a starting concentration of around 50,000 mites •per litre.

According to the present invention. there is provided a composition comprising at least one predatory mite, and Thyreophagus entomophagus as a food source for said predatory mite.

In one embodiment, the composition is self-sustallring. It includes a food source for the prey mite, which in turn is a food source for the predator mite. In one aspect of


the invention, the composition comprises a population of predatory mites. The population may be a breeding population so that the composition provides a continuous supply of predatory mites. In another aspect of the invention, the

Thyreophagu.s l!lliomophagu.r is a population of prey mites. Again, this population may be a bree<ling population so that the composition provides a continuous !>Upply of prey mites as food for the predatory mites. In a further aspect of the invention) the composition may comprise other food sources for the predatory mites in ~ti.on to the Thyre"fJhagu& enlon•"flhagu& prey mites. Other food sources may include natural prey of the pred!rtors such as Tetranych:us urticae for the predator Amblyseius swirskii, honeydew, baker's yeast or pollen. The pollen maybe from m:LY suitable source, such as the date palm plant Phoenix dacty!ifera, or the castor oil plant Ricinus commurrit.

The composition may be employed as a means for rearing large numbers of predator mites. In one embodiment, the composition is used as a means of rearing large numbers of Amblyseius swirsldi predator mites. Alternatively, the composition may be packaged in such a way that it can be placed directly in a crop environment Since the composition is self-sustaining, it can be placed directly into a cropping environment where it can be used to provide a continuous source ofpredator mites for a prolonged period oftime. In one embodiment, the composition will be self-sustaining for at least a week. In another embodiment, the composition will be self-sustaining for a least a month. In a further embodiment, the composition will be self~ sustaining for 6 weeks or more. In one aspect of the invention, the ratio of predator to prey mites in the composition will be adjusted to ensure tltat the rate of supply of predator mites remains approximately constant for a period of at least a week. In one aspect of the invention, the ratio ofpredator to prey mites .in the composition will be adjusted to ensure tltat the rate of supply of predator mites remains approximaiely constant for a period of at least a month. In a further aspect of the invention, the ratio of predator to preynlltes in the composition will be adjusted to ensure that the rate of supply ofpredator mites remains approximately constant for a period of 6 weeks or more.

Suitable cropping environments include, hut are not limited to, glasshouses, greenhouses, polytunnels, shade houses (for example netting structures used for

ornameotals and licld grown crops ofpeppers and tomatoes), orchards, fields (for example for strawberry and raspbetry crops) and gardens (for example market gardens and botanical gardens).

The composition may be delivered to the crop via any suitable delivery system, ranging from simple devices such as pots, bottles, boxes, cartons, bags, tubes and sachem to more complex devices such as the Geminin>~ twin sachet (as described in

GB2393890), blowing devices which carry product onto lile crop in a strearu of air or liquid, and rotary devices which distribute product mechanically into the crop. Preferably, the delivery devices will include a means for the predator mites to be released from the composition onto the crop. Such means may be in the form ofone or more em6I'gence holes. Preferably, the delivery device is designed or otherwise placed in the crop environment such 1hat the composition inside the device will not get wet upon rainfall or watering of the crop. Depending on the design of the delivery devioe, it may be suspended from or biDlg an branches throughout 1he crop so that the predator mites are released at regular spatial intervals throughout the entire crop.

In one aspect ofthe invention, the mite composition will be sprinkled as a loose product onto the crop from bottles. In another aspect of the invention, the mite composition will be placed in the crop in sachets or release boxes. In a further aspect of the invention, the mite composition will be blown into the crop using handheld or motor-drivcm blowers, tractor-mounted delivery sy.ste:ms, systems which drop dry product onto a spinning disc, or systems that sprinkle product from bottles or flasks attached to a rotating mm.

In one aspect of the inven~on, the composition further comprises a food source for

Thyreopha?;JJS entomopluzgus. '!none embodiment, the food source is a natural diet for Thyreophagus entomoplw.?;JJS. Since Thyreophagus entomophagus is a grain mite1
the food source may be derived from grain. In one embodiment, the food source comprises bran, rice hulls, rolled oats, com grits, flour (such as gram flour, buckwheat flour or cereal flours), dried fruit,j~ dried insects or poultry meal. In another embodiment, the food source is an artificial diet. A carrier material such as buckwheat husks may also be present

In a further aspect of the invention, the .food source for Thyreophagus entamophagus is sugar-rich. The term "sugar-rich diet" in this context is defined as one that contains at least 5% sugar. A sugar-rich diet is useful for rearing prey mites that sra more palalable for predator mites. Predator mites favour prey mites fed on sugar-rich diets compared to those reared on low-sugar diets. Therefore prey mites reared on sugar-rich diets provide a better food source for predator mites . .AB a result, predator mites are healthier and less frtressed, leading to increased egg production and the production of a greater number of predators.

Matsumoto, K (lap. J. Sanit. Zoo!. 15, 17-24; Jap. J. Sanit Zoo]. 16, 86-89; and Jap. J.

Sanit. Zoo!. I 6, I 18-122) discloses that Carpoglyphus lactis uses glucose and sucrose, but not stmch, and that its maximum rate of increase is achieved when reared on a diet of 60% yeast and 40% sugar. Matsumoto sJso discloses that Iyrophagus putre.scentiae can metabolise a. variety ofcarbohydrates, but tha:t it reaches its maximum rate of increase on pure dried yeast Further~ it describes that in mixed populatiODS, C. lactis predominates in diets containing up to 40% sugar, and T.

putresce.nti.ae predominates in pure dried yeast.

In a further aspect of the invention, the food source for Tkyreophagus entomophagus is low in starch content. A low starch content is less than 30% starch. In a further aspect of the invention, the predatory mite is Anzb!yseiu.r swirs!di. A sugar-rich and/or low-staroh food source may be used in conjunction with the present invention in any of the methods for rearing predatory mites, or compositions described above.

In one ernbodimen~ the food source is a yeast-baaed artificial diet. In another embodiment) the :fuod source contains simple sugars. Simple sugars include those
•    such as allose, altose, dextrose, glucose, sucrose, nUIDilose, gulose, idose, galactose, talose, fructose, saccharose, lactose and arabinose. Preferably, the simple sugar in the diet is dextrose. For example, the diet may comprise 5%, 10%, 15%,20%,25%, 30%, 40%, 50%, 60% or more than 60% simple sugars. In a further embodiment, the food source has a low starCh content, for example less than 30%, suitably less than 20%, more suitably less than 10%, more suitably still less than 5%, most suitably less than


1% sta:rolr..  In one en1bodiment, there is more sugar .in the diet than starch. ID a

further embodiment, the ratio of simple sugars to starch in the diet ranges from 1:1 to

1000;1. Sllitablytheratio of simple sugars to starch in the dietr.mges from 2:1 to

10:1.

A sugar~rich diet is useful for rearing Thyreophagu.r entonzopluJgu.r because this mite

is able to metaholise simple sugars. Many other mites, such as Tyrophagus

putre/Jcentiae arui Acarus •iro fuvour lltarcil-rich diots and lack a direct ability to

u.tili.se simple sugars in their diet.  In contrast, Thyreopha~ entomophagus can

utilise both simple sugars and starch (Al<imov et al. Elcologiya (Ekaterinburg, Russian

Federation) 2, 27-31).  Diets that are rich in starch rapidly become infested with T.

putrescentiae, which rapidly ovenvhelms other mites such as Thyreophagus entomophagus. Therefore, a diet ]ow in starch and rich msimple sugars is useful to reduce or even prevent the onset of contamination by other mite species. T. pu.trescentiae does not thrive on low starch diets because it needs to seareh for food,
and the starcil food runs out quicldy.

Optionnlly, the diet may additionally contain other food sources that are desirable for

the predator andJor prey mites to feed on. For example, the diet may contain pollen

such as that from Typha sp., date pahn plant Phoenix dactylifera, or castor oil plant

Ricinu.s communis.

Non-limiting examples of possible diet recipes that can be used in accordance with the present invention include: i) 60% yeast1 20% wheat germ and 20% fish food; ii) 50% yeast and 50% dextrose; iii) 55% yeast1 25% dextrose, 10% wheat ge:rm and 10% fiB1I

food;  I1Jld iv) 30% yeast, 30% dextrose, 20% wheat germ, and 20% soya flour.  The

present invention includes obvious variants of these diets that would be readily

contemplated by the person skilled in the art.

According to the present invention, there is provided a method for controlling pests in

a crop comprising providing a predator 1nite that has been reared using Thyreophagus

entomaphagus as a host  In one embodiment. the predator mite is Amblyseius sV~rirskii,

and the pests are thrips and/br whitefly. Amblyseius swirskii is a preferred predator

tnite sinee it feeds on either thrips or whltefly. Both thrips and whitefly are major pests of crops, especially in closed cropping envi!omnents such as glasshouses. Using
Ambly<eilM swirskii, both of these crop pests can be controlled using a single biological control product. The present invention is used for controlling any crop pests, such as one or more listed above. Further tb.e present invention is used in

conjl.ID.ction with any crop, such. as one or mare offuose l.isterl above.  Suitably, the

crop is select.ed from the group consisting of peppers. cucumbers, aubergines, roses,

ge.rbEtt'as, melons and bea.ns.

Acconl.ing to the present invention, there iB provided the uso of Thyreophagus entmnophagus as a rearing host for predatory mites. In one aspect of the invention,
Thpeophagus entomophllgus is used as a rearing host for Amblyseiu.s swirslrii,

Ar.conling to tho present invention, there is provided a method of rearing mites comprising providini a sugar-rich food source, and allowmg the mites to feed on said food somce. In one aspect of the invention, the food source is low in starch content

In another aspect of the invention, tb.e food source comprises simple sugars. Simple sugars include those such as allose, altose, dextrose, glucose, sucrose, :ma:rm.ose, gulose, idose, galactose, talose, fructose, saccharose, lalrtose and arabinose. Suitably, the food source comprises dextrose. This method oan be used to rear 11llY mites that are able to metabol\se sugars. In a further aspect of the invention, the mites to be reared are Thyreuphagus entomophagus.

Ar.conling to the present invention, there iB provided a mite food source tbnt is high in sugar and low in starch content. In one aspect of the invention, the mite food source is derived from yeast. Suitably the yeast is brewers yeast. bakers yeast, molasses yeast or lactic yeast. More suitably, the yeast is brewers yeast The mite food source is particularly suitable for rearing Thyreophagu.s entomophagus, and other prey mites tbnt metabolise sugars such as Carpoglyphw lacru, Glyc:yphagus destructor, G.
domesticus, G. onwtL~E1 G. geniculatus, CtenoglyplUtS plumiger and Su.idasia

meda.nensis,  In particular, the mite food source is suitable for rearing Thyreo]ihagus

entomophagus andlor Carpoglyphw; lactis.  The mite food source is also used for

rearing prey mites to support a breeding pC>pulation of predator mites, such as

Amblyseius swirskii.


EXAMPLES

Example 1 : Comparative productivity of Amblweius ~trslcii when reared on two species: of prey mite. CarpqglJi,phus lac.t:is or Thvreoohagus entomophaoo

1.1  Background

A series of cultures ofAmblyseiw (Tjph/odromips) swtrsldi were setup an two different prey mites, Thyreophagti.s entomophagus and Carpog/yplws /actis. The food

sml!ce for the prey mites was a 50:50 mixture of brewers yeast and dextrose for all

cultures excePt cultures 3 and 8 using Thyreophagus entomophagus as prey mites.

The Thyreophagus entomophagus prey mites in cultures 3 and 8 were reared on a diet without dextrose, and incoxporating fish meal as a source of protein. 1n all cases, the
food was mixed with a carrier material.

T.be number of predator mites per litre of culture material in late stage cultures was counted. Counts were made acccm:Ung to a dry counting method previously used as a standard byMAFF (Griffiths eta/. 1976, Ann. App. Bioi. 82, 180-185). This method allows multiple samples to be assessed within a. short period of time, although it is recognised that it does not recover all of the mites in a sample.

In total, there were 34 cultures using ThyreophaguJ' entomphagus a.s prey mite, and 56 cultures using Carpoglyphus /actis.

1.2  Culture density

Table 1 below indicates the average density of A. swir~ldi per litre of culture material.

TABLE!

Number of A. swirskii per litre of culture material, using;

Thyreophagus as prey mite    Ca1poglyphus as prey mite
    1    150,000        115,000       
                           
    2    120,000        90,000       
                           
    3    40,ooo•        100,000       
                           
    4    100,000        90,000       
    5    150,000        100,000       
                           
    6    160,000        50,000       
    7    120,000        90,000       
                       
                           
    8    20,000*        100,000       
                       
                           
    9    65,000        90,000       
                           
    10    100,000        50,000       
                           
    11    90,000        120,000       
                           
    12    100,000        65,000       
                           
    13    90,000        80,000       
                           
    14    150,000        1,00,000       
                           
        IS    100,000        95,000       
                           
                           
    16    80,000        8,0,000       
                           
    17    100,000        100,000       
                           
    18    100,000        80,000       
                           
                           
    19    100,000        50,000       
                           
    20    60,000        60,000       
                           
    21    80,000        90,000       
                           
    22    60,000        85,000       
    23    80,000        150,000       
                           
                           
    24    75,000        80,000       
                           
    25    90,000        80,000       
                       
                           
    26    90,000        56,000       
                           
    27    80,000        64,000       
                           
    28    75,000        65,000       
                       
                           
    29    80,000        80,000       
                       
                           
    30    100,000        80,000       
                           
                           
    31    80,000        20,000       
                           
    32    80,000        75,000       
                       
   

            33        100,000        20,000       
                                   
                                   
            34        100,000        65,000       
                                   
            35        nla    75,000       
                               
            36        nla        65,000       
                                   
            37        nla    20,000       
                               
                                   
            38        nla    75,000       
                               
            39        nla    75,000       
                               
            40        nla    65,000       
                               
                               
            41        nla    65,000       
                                   
            42        nla    20,000       
                               
            43        nla    40,000       
                           
        44        n/a    60,000       
                           
                                   
        45        n/a    65,000       
                                   
        46        nla    60,000       
                           
                           
        47        n/a    20,000       
                           
        48        n/a    40,000       
        49        nla    65,000       
                                   
        50        nla    60,000       
                           
        51        n/a    50,000       
                       
    52        nla    65,000       
                       
    53        nla    130,000       
    54        n/a    65,000       
                       
                                   
    55        n/a    40,000       
                       
    56        n/a    20,000       
                           
        Average (mean)    93,088        70,625       
                                   
"' Prey m1tes reared on dtet wtthout dext:Iose

The results indicate that, on average, 31.81% more predator mites were produced by

rearing on Thyreophagu. than on Carpogl)phus.  A statistical analysis of the results

was made using the "t tesf'. ass:un;ring equal variances incorporated in the Data

Analysis piU:kage in Micr080ft Excel.  This result was found to be statistically

significant, having a P value of0.006394.

The Thyreophagus mites in cultures 3 and 8 were reared on a different diet to all othor

cultures, namely one that did not contain dextrose. The number of predator mites

produced when reared on Thyreophagus is not as high when the prey mite is fed on a

dextrose-free diet than on a diet containing dextrose. When Thyreophagus reared on cultures 3 and 8 are excluded from the analysis of the data in table 1, the mean number ofA. swirskii mites produced on Th.yreophagu.s cultures is 97,031, and on

Carpoglyphus was 70,625.  This indicates that, comp&ing produotion on constant

diets, on average 37 39% more predator mites were produced by rearing on

Thyreophagus than on Carpoglyphus. This result is statistically significant, having a

P value of0.00()(JI597.

L3 Population Growth

The culture batches from ex&nple 1.1 were tracked throughout the later part of the culture developmont cycle. At 5 different culture stages, the number of Amblyseius

swirski.i predator mites was counted in each culture by the sampling method described

above.  The 5 culture stages are defined by the number of weeks after starting the

culture.  Stage 1 is culture week 8, and has a total culture volume of approximately 2

Jitres.  Stage 2 iB culture week 9, and on average has a total culture volume of

approximately 6 litres. Stage 3 is culture week 10, and on average has a total culture

volume of approximately 151itres.  Stage 4 is culture week 11, and on average has a

total culture volllllle of approximaJely 45 litres. Stage 5 is culture week 12, and on average has a total culture volume of approximately 135 lill'Os.

It is important to note when interpreting these figures that cultures are started in low

volumes of carrier mate:rial with relatively small numbers of mites.  As the mite

population density increases, the culture volume is increased to maintain a relatively

constant mite density.  These figures are obtained by multiplying ti1e density recorded

at each stage by the volume of culture which exists.  Counts are not made at earlier

stages of the culture (i.e. before culture week 8) because the disturbance involved in the small volumes of culture which exist at those stages would adversely affect mite petfonnance and colony growth. Therefore, even at culture stage l, the number of

predator mites bas already been influenced by the t)lpe ofprey mite available, ami the nature of the food supplied to that prey mite.

TABLE 2 - this includes all culture batches referred to in Table 1

    Culture stage    Actual yield of A. swirsldi (thousands of mites)   
        Thyreophagus    Carpoglyphus   
               
               
    1    189.00    1!9.27   
               
    2    563.33    467.40   
               
    3    1,218.75    1,275.00   
               
    4    3,085.71    3,318.75   
           
5    11,250.00    9,642.86   
               

TABLE 3- this includes ali culture balches referred to in Table 1 except

Thyreophagus culture batches 3 and 8, having no sugars in diet

Culture stage    Actual yield of A. swirskii (thousands of mites)
       
    Thyreophagus    Carpoglyphus
       
1    188.75    119.27
       
2    552.86    467.40
       
3    1,475.00    1,275.00
       
4    4,320.00    3,318.75
       
5    16,875.00    9,642.86
       

The data shows that cultures of Amblyseius swirskii reach significantly higher average

densities when fed on Thyreophagus enJomophagus than when fed on Caipoglyphus

lacti.s.  Moreover, cultures reared on T. entomophagus with dietary sugar perform

better than those reared on T. entomophagus without dietary sugar. Thus both the

species of prey mite offered, and the diet on which that prey mite is reared, influence

the final density of the cultures achieved.


Example 2: Rearing ofAmblvseius cucumeris on Thvreophagus entomophagus

A small culture of Thyreophagus entomophogu.r was found to be contaminated with

large numbers of the predatory mite Amblyseius (Neoseiulus) cucumeris. The source

of this corrtamination was unknown. Due to the lBrge numbers of A. cucu.meria visible,

the culture was counted. Four thousand A. cucumeris were present in the culture,

whiohhild a volume of24cm'. Tbis is equivalent to adBDSity of 167,000mites per litre, which exceeds typical production densitie.e of 110,000 mites/litre for A.
cucumeris fed on Tyrophagus putrescentiae.

The culture was kept and expanded. Initially, the culture was transferred to a 265cm' fla.sl:, with the eddition of 40cm' ofT. entomophagu3 culture, 80 em' of fresh bran, and 1Ocm' of food for the prey mites (a 50:50 mix of brewers yeast aJJ.d dexlrose),

giving an approximate total volume of 154cm' of culture medium. S days later the

culture flask was examined, and seen to contain very high mite densities. It was then traosferred to a 650cm' flasl~ with the eddition of an additional 20cm' of fresh bran. Nine days later, the culture was counted Jllld fuund to contain 40,000 Amblyaeius cucwneris. Thns, the total population increased 10 fold within 14 days, and the density of A. C!lcwneris rose to 230,000 per litre. '!'biB is more than double the density

which is routinely achieved fot A. cucumeris during more than 18 years of commercial production of this predator on a diet of Tyrophagus putrescentiae.

Example 3: Testing the effects oftbesoecies ofpreymheoffered. and the food given to that prey mite on the productivity of the predatory mite Amblyseius
(TJphlodron'lips) swt.rildi

3.1 Baclrground

A trial was set up to assess the influence of prey mite species and prey mite food type,

on the production of the predatory mite Amblyaeil/8 (I'yphlodromips) swirakii. Three food mixtures were used fur this trial, as shown in tsble 4.

TABLE4

Diet A

A 50:50 mix by volume of each of the individJial food blends with bran was placed

it.to Eppendorf®  tubes with the lids removed. The individual tubes were each loaded

with 10 reproductive prey mite famales and one male, then plugged with cotton wool

and placed upright in. racks.

Tubes containing the mite Carpoglyphus lactis were placed at an ambient relative

humidity of75% and a temperature of24'C. Higher humidity resulted in a complete

deliquescence of the-food in previous trials with C. lacti.s.  Tubes containing

Thyreoph.agu.s entomoph.agus were placed into closed polypropylene boxes, and a relative humidity of 80% maintained by a solution of glycerol in wa±.er.

Mite populations were allowed to develop for four weelcs, and fed fresh food after 2

weeks to maintain population growth  At the end of four weeks, an individJial female

ofAmblyseius (Typhlodrorrrips) swirskii was placed into each tube, and the tube re-

plugged with cotton wool A:fter a further seven days, tubes were opened and the

numbers ofA. swirskii eggs, juvenile8 and adults counted. For Thyreophagu.s

entomophagus tests, five replicates were perfonned, and for Carpoglyphus lactis, I 0

replicates were perfurmed.

3.2 Results

The average (mean of all replicates) numbers of eggs, juvemiies and adu1ts is given in

TableS.

TABLE 5: Meao results (average numbers of eggs, juveniles and adults)

                               
    Thyreophagus entomophagus        Carpoglyphus lacru   
                               
    Eggs    Juveni!CB    Adults    Total    Eggs    Juveniles    Adults    Total
Diet A    1    3.6    3.2    7.8    0.3    2.7    0.6    3.6
DietB    0.4    0.2    I    1.6        -    -    -
DietC    0.8    2.8    3.2    6.8    1.3    1.9    1    4.2
                                               

Both diets containing dextrose (A and C) produced substantial increases in the =bets of predatory mites over the course of the trial. On diet A, Thyreophagus en.tomophagus produced more than double the number of A. swirskii than did

Carpoglyphus lacti.s fed on the same diet. On diet C, Thyreophagus entomophagus produced 38% more A. swirskii than did Carpoglyphus lactis fed on the same diet.
The presence of additional sources of protein in diet B resulted in poor mite

reprodnction, and in the case of Carpoglyphus lacti.!, the mites performed so poorly

on this diet that no A. swirskii were introduced.

The results were statistically analysed using Students t test (from the statistical analysis tools included with Microsoft Excel), as shown in tables 6 and 7.

TABLE 6: Compadson of different food blends

        Diet A vs Diet B    Diet A vs Diet C    Diet B vs Diet C
               
    T. entomophagus    0.002607**    0.243641    0.000265**'
               
    C.lacti.!    nla    0.34226!    nla
               
    TABLE 7: Comparison of different prey mite species   
           
            T. entomophagus v C. lactis
               
    Diet A        0.01374*   
               
    DietB        nla   
               
    DietC        0.022953*   
               
n!a- companson not possible due to lack of data.

"' Signific.mt difference

**Highly sigoificant difference

"'"'"' Very highly significant difference

The results clearly show that the presence of dextrose in the diet gives very significant improvements in productivity of A. swirskii fed on either prey mite species. Further, when given the same diet, Thyreophagus entomopluzgus is supclior to Carpog~phus la.cti.s as a prey mite for rearing Amblyseius ~irskii.


24
 


25

CLAIMS


1. A method for rearing predatory mites, comprising providing Thyreophagus entomophagus as prey mites, and allowing the predatory mites to feed on said prey
s    mites.



2. A method according to claim 1, wherein the prey mites are present as a breeding population.

10    3. A method according to claim 2, wherein the predatory mites are from the family Phytoseiidae.


4.  A method according to claim 3, wherein the predatory mites are from a genus

selected from the group consisting of Amblyseius, Typhlodromips, Neoseiulus.

1.5    TyphlodromaJus. Euseius, Typhlodromus,lphiseius and Kampimodromus ..


5. A method according to claim 4, wherein the predatory mites are selected from the group consisting of Amblyseius andersoni, Amblyseius ca/ifomicus, Amblyseius cucumeris, Amblyseiusfallacis, Amblyseius limonicus, Amblyseius montdorensis,

20 A.mblyseius ova/is, Amblyseius stipu/atus, Amblyseiu.s swirsldi and Amblyseius womersleyi.


6. A method according to claim 5, wherein the predatory mites are Amblyseius swirskii.

25

7. A composition comprising at least one predatory mite. and Thyreophagus entomophagus as a food source for said predatory mite.

8.  A composition according to claim 7. further comprising a food source for
 

30    Thyreophagus entomophagus.
 

26

9. A composition according to claim 8, wherein the food source for Thyreophagus entomophagus is sugar-rich.

10.  A composition according to claim 9, wherein the food source for Thyreophagus

.s    entomophagus is Iow in starch content.


11.  A composition according to any one of claims 7 to 10, wherein the predatory mite

is Amblyseius swirskii.


10 12. A method for controlling pests in a crop comprising providing a predator mite that has been reared using Thyreophagus entomophagus as a host.


13. A method according to claim 12, wberein the predator mite is Amblyseius swirskii, and the pests are thrips and/or whitefly.

14. A method according to claim 13, wherein the crop is selected from the group consisting of peppers, cucumbers, aubergines, roses, gerberas, melons and beans.

15. Use ofThyreophagus entomophagus as a rearing host for predatory mites.

20

16. Use of Thyreophagus enromophagus according to claim 15, as a rearing host for

Amblyseius swirskii.


1'7. A method ofrearing mites comprising providing a sugar-rich food source, and

2.S    atlowing the mites to feed on said food source.


18. A method according to claim 17, wherein said food source is low in starch

content.


30    19. A method according to claim 18, wherein said food source comprises simple sugars.
 


27

20.  A method according to any one of claims 17 to 19, wherein the mite is

Thyreophagus entomophagus.


21 . A method according to any one of claims 17 to 19, wherein the mite is

s    Carpoglyphus lactis.
 


INTERNATIONAL SEARCH REPORT    llnlernaUonalappUcatiDnNo   
    J PCT /GB2007 /002858.   
       


ACCO!dnglollllefnaliOniiPalentCiasBiriCalion{IPC)orlobolllnattonaldaSsllicallol'landiPC

    B. FIELDS SEARCHED                                   
    Minimum documentalion sell;ldled (classificallon syatem followed by ciBSS!II:allon synt!ols)       
    AOIK    A01N                                   
                   
    Dowmenli!lion searched olher than nunimum doc;umentatian to !he extent that Sllch documants are Included In the f~elds searched   
                       
    EleCironlcdlllabase.(Dilsulledduring the  lniBmalionalsaiii'C!l(name•~ldalabaSeand, where prad~~rcii\Brmsused),       
    .EPO-Internal, •WPI Data                               
                               
    C. DPCUMENTS CONSIDERED TO BE RELEVANT                       
    CategDry•    c•allonofe!O!:Ument,wllhindicabOn,  whereapprupriBLe,oltheralevanlpiiSlillgeS        Relevant to daim No.   
                               
        WO. 2006/057552    A (KOPPERT BV    [NL];        12-14   
        BOLCKMANS KAREL JOZEF  FLORENT  [BE];  HOUTEN           
        YVONNE    MAR) •1 June 2006    (2006-06-01)           
        clted    :in  the  app1icat1on                       
        cl•lms    1,18        •                       
        ZHANG    Y ET AL:        "Potential  of    amblyseius        7-11   
        cucumeri s  (Acari:  phytosei idae)  as  a           
        biocontrol•agent    against    schizotetranychus           
        nanjingensis  i"n    Fujian,    China"           
        SYST£MATIC AND APPLIED ACAROLOGY •SPECIAL           
        PUBLICATIONS,    SYSTEMATIC    AND    APPLIED           
        ACAROLOGY SOCIETY,. LONDON,  GB,           
        vol •  4,  2000,    pages  109-124,    XP003002569           
        ISSN:    1461-0183            .               
        abstract                               
                            -1--           
                           
    [!] FlalhardocumentsaraiiSIBdtnthe c:onlinUilllorl_oiBni.C.                   
                                   
    •  SpecialcalegorissolcileddoalmB!11S:                "T' lalefdoclltnilnlpubll!ltlBdllllariMI inll!mali~;~nalftll~~gdale   
                               
    'A" dorurNntdellnlngthege~~eral!!lale olllle artwhlchisnlll            ~;~rpriml!yd!deandn~;~tln~ntbctwllhlheappllcaUonbUI   
                clllld\clunclers;landllleprlnciJ*orlhe~;~ryWldlll"lojrlglhe   
    consldaredlcbeofpartbllarrulellanc:e                lnvanlloo       
    "E'aariardor;:urrsntbutpqbllshsdonoranerlllBillll!malional        "X"docllmlnlolpalllr::ularral!M!nc:e;lhecllli!nBditlV!IntiOn   
    IDingtlalll                           
                            cannalbec:on!ilderadnOY!!IorcannolbeCOII!IIdl!tlldl~;~   
    "L' document which may lhrow !Dubls~n pri:uity diDn(l;) or            nvoM!anPvenltffllllepVIflenthedoCI.Imenlls takellalone   
    whChisclledloasliii!IIShlllePU~hc:allondaleolanothar        •r doculliEIIIlofj:BIIIcullltral!MIIlce:1heclaim&d    bwllfllion   
    dlallonoroDlersped!!olt81150n(as •spedflld)               
                CBnootbeCOilSideiEidto'JwolveanlnVBnliv'B&lepwhenlhe   
    '0'  dooumel!lreleningtoanorllldisdosure.u!IB, axhibDionor            dDCUtTIBflli&mrrillnedwllhoneormoremher suchdtlcu-   
    "olhBrt!IBIIIIS                        mslll&,suchoomblnallonbelngotwll)llll\oa    p&IIIOnskllled   
    "f"docutnel'ltpub!ishedprtortolheinlemallonalfilingdalebul        lnthesn.       
    lalerlhanlhepriorllydatec8rlll!d                    '&" dotumEIII\tTIIItlbBtoflhBSi!llfl!!pai!JIIIfam)ly       
                   
    Daleoflheadualcompletlonoflhelntemallonalaeiltttl            Dalellflmi.Hilgoftheinlemallohalseard!ltlport   
    2 April  2008                    23/04/2008           
                                   
    NameandmalllngaddressoltheiSAf                               
        europeanPaLeii!Oftiee,P.B.5B1BPatlltlllaan2                   
        NL-2280HVRijswlj'K                               
        Tet(.. 31-70)J4D-2040,n.3t6t;1apon~            Pl 11 e,  Stefa an       
        Fa.r:(+31-70)J40-3016                           
                                   
    FoiMFCT/15A121D(S8CC11"1d-)(Aprli20DS)'"                               
                                    page  1 of 2   
INTERNATIONAL SEARCH REPORT    lntematlonlll applk:atlon No   
    lPCT /GB2007 /002858   
       
C(Conllnuatlon).    DOCUMENTS CONSIDERED TO BE RELEVANT       

Calegory-   'CilatkinoiOOaJm(ll'll,wllhlndlcation,whereappropl'tala,ollhemlevar:'lp&ssaQIIS    FlelevantloclalmNo

A    ZDARKOVA E:    "Mites  as    pests  of stoced    7-11   
    products.  14.1.  Application  of the       
    bio-preparation  'cheyletin'  in  empty       
    stores"               
    MODERN AGCAROLOGY,  ACADEMICA PRAGUE AND       
    SPB ACADEMIC  PUBLISHING,  THE  HAGUE,  NL,       
    vol.  I,  1991,  pages  607-610,  XPD03DI4954       
    abstract               
    RAMAKERS P J    M ET AL:    "Start  of    7-11   
    c•ammercial  production  and -introduction  of       
    amblyseius  mck.enziei  sch.  & pr.  cacar1na"       
    MEDEDELINGEN VAN DE FACULTEIT       
    LANDBOUWWETENSCHAPPEN    UNIVERSITE IT GENT,       
    GENT;  BE,               
    vol.  47,  no.  2,  1962,  pages  541-545,       
    XP003D00199               
    ISSN;  0366-9697           
    abstract               
    WO 99/59402 A (PREDATION  INC  [US])    22,23   
    25 November 1999 (1999-1!-25)       
    page 6,  line 14;  claims  1;12       
    ABOU-AWAD B A ET AL:    "EFFECTS OF    22   
    ARTIFICIAL AND NATURAL DIETS  ON THE       
    DEVELOPMENT AND REPRODUCTION  OF TWO       
    PHYTOSEIJD MITES AMBLYSEIUS  GOSS1PI  AND       
    AMBLYSEJUS SWIRSKIJ  (ACARI:  PHYTOSEIJDAE)"       
    INSECT SCIENCE AND  ITS APPLICATION,  ICIPE       
    SCIENCE  PRESS,  NAIROBI,  KE,  .       
    vol.  1>,  no.  3,  1992,  pages  441~445,       
    XP009050376               
    ISSN:  0191-9040           
    abstract            23   
                   
    GB 2 393 B90 A (SYNGENTA B!OLINE  LTD  [GB])    23   
    14 April  2004  (2004-04-14)       
    page  1,  par'agraph I           
    page  10,  line 4- line I!       


    1    ln18rn~onal applicaijon No.   
INTERNATIOKAL SEARCH REPORT        PCT /GB2007 /002858   

Box No. II    Otnu"I'VIItionliwhen! cerlan claims wen! found unsean::hable (Conti~aHon c;~l ttem 2 offlrslsneet)

This lntsmatlonal Saait:h report has not been "esta.DtiShed hi respect of cef!aln claims under Artide 17(2)(a) tor the lollowi~ rea!lons:
.    .    .           
1.  [i] ~!1¥1NO~.:    1-6    15-21    .  •    :
because they relate to sub)ec:t    ~atter not required to be ~ardlac:l by this Authority, namely:

Rule 39.1(ii) PCT- Essentially biological process for the production of animals

2.  OclalmsNos.:
because they relate to parts ot the international appliCation lhat do nO! comply With lhe prescribed requirements to such an extent that no meaningful international seardl can be carTied out, spadl!cally:


3.  Oe~aamsNos.:    .    .    ,
because they are depanaent claims an_d are not drafted in ac=ordance wtlh the oocond and l'ntrd sentences of Rule 6.4(a).

Bor: No. Ill    Observations where unity of invention is la::king .(Continuatloo of Item 3 of llrst sheet)

This International Ssarchlng Authority fotJ~ multipl!llrwenllons IIi this lntamaUonal ap~icallon, as follows;

see  additional  sheet


1.    GJ :.~s~lred additional seait:h fees we~ timely paid by ih_~ appRcant, this lnlematlonal seout:h report covers aosea.tha.Die

2.    D    1>s au searchable cl~s could bEi searched wllh~ut errort jusUfylng an additional fees, this Authority did notln\llle payment of

addlllonal fees.

<1. D No re(J.Iired additional search fees were timely paid by the appllcanl ConseQUently, lhls international search report is restricted to the Invention first mentioned in the claims; II is covered by ClaimS Nos.:

Remark-on Protest    TI)e adliflonal search tees were accompanied by lhe applicanrs pr01est and, where applicable, the
    D paymenlofeprotestlee.
    DThe additional search lees were accompanied by lhe applicanrs p_rotest but the appllcab~ protest
    fee was not paid within lhe lime limit spad~ed in tile Invitation.
    r::J No prolest accompanied the payment of adc:litiqnal search lees.

Form PCTIISA/210 (continuation of first sheet {2)) (AprU 2005)

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