EP3550970A1

EP3550970A1 - Kit of attractants for attracting the coffee berry borer

Info

Publication number: EP3550970A1
Application number: EP17822367.3A
Authority: EP
Inventors: Brigitte FREROT; Stéphane DESQUILBET
Original assignee: Institut National de la Recherche Agronomique INRA
Current assignee: Institut National de la Recherche Agronomique INRA
Priority date: 2016-12-06
Filing date: 2017-12-01
Publication date: 2019-10-16
Also published as: WO2018104625A1; US20190307123A1; FR3059518A1

Abstract

The present invention relates to a kit of attractants for attracting the coffee berry borer, Hypothenemus hampei, comprising a mixture of ethanol/methanol and (-) limonene and/or 2-ethylhexanol, as well as to the uses thereof and methods employing said kit.

Description

ATTRACTIVE KIT TO ATTRACT COFFEE SCOLYTE

FIELD OF THE INVENTION The present invention relates to an attractive composition and its use in trapping the coffee borer Hypothenemus hampei.

BACKGROUND OF THE INVENTION Coffee borer, Hypothenemus hampei (Ferrari), in English Coffee Berry Borer, is the most damaging pest in coffee growing. Originally from Africa, the coffee bark beetle has colonized almost all coffee producing regions of the world (with the exception of a few islands) and has resisted eradication programs. This pest causes on the one hand important losses (up to 90% infested cherries in untreated plantations), and on the other hand affects the quality of the grains.

The cryptogamic lifestyle of the insect and the appearance of resistance make the chemical treatments not very effective in the fight against this pest. In addition, agricultural policies tend to move towards alternative solutions to the use of pesticides. The search for biological control methods against coffee borer is therefore a major issue for coffee producers.

In this context, several studies have been carried out since the 1990s and have highlighted the attractiveness of coffee cherries' scents on bark beetles. In addition, mass trapping appears to be a promising biological control method for controlling this pest. Indeed, bark beetles have a coffee-dependent life cycle, reproduction can only be done in cherries whose coffee beans are already formed (100 to 150 days after flowering). After harvests, a period begins when breeding-breeding kernels become scarce and females known as colonizers must leave the dried fruit in search of new cherries at the next fruiting stage. It is at the end of this phase of survival that mass trapping would be the most relevant.

More particularly, studies have demonstrated the efficacy of an ethanol / methanol mixture as an attractant for the mass trapping of coffee beetles (see, in particular, A. Ortiz et al, Journal of Agricultural and Food Chemistry, Vol 52 19, 5914-5918, 2004). Nevertheless, the improvement of this attractant with a good specificity for the coffee borer is of major interest to be able to further limit the infestation of coffee borer, and thus increase the yields and the quality of the coffee crops. SUMMARY OF THE INVENTION

The inventors of the present application have managed to develop a more effective attractant, based on the addition of volatile organic compounds emitted by coffee beans and likely to intervene in the recognition of host plants by bark beetles. More specifically, the inventors of the present application have discovered the advantage of adding to the mixture of alcohols used to date, a molecule of the terpenoids class: (-) limonene and / or a particular branched alcohol, the 2 -éthylhexanol. This discovery is particularly unexpected in that 2-ethylhexanol is rarely emitted by plants and was first demonstrated by the Applicants as constituting volatile organic compounds emitted by coffee cherries. Similarly, if limonene was known to constitute coffee beans (E. Naptali et al., Http://www.scielo.org.mx/pdf/jmcs/v57n4/v57n4a9.pdf), the (-) limonene has never been considered as attractive to the bark beetle, even though several volatile organic compounds emitted by coffee cherries have been proposed for this purpose (BP Dufour et al, International Scientific Symposium on Coffee, ASIC, Vol 24, 922 -926, 2012).

Thus, the present invention relates to a kit of attractants for attracting the coffee borer Hypothenemus hampei, characterized in that it comprises a mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol.

The invention also relates to a device comprising the kit of attractants according to the invention, said device being capable of trapping and / or killing said bark beetle.

Thus, the present invention relates to the use of the attractant kit or device according to the present invention for attracting the coffee borer Hypothenemus hampei, preferably for trapping and / or killing said borer or for modifying the behavior of said borer.

It also relates to a method for attracting the coffee borer Hypothenemus hampei, in which the kit of attractants or the device according to the invention is placed in or near an attraction site. DETAILED DESCRIPTION OF THE INVENTION

Attractive kit

According to a first aspect, the present invention relates to a kit of attractants for attracting the coffee borer Hypothenemus hampei, characterized in that it comprises a mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol .

The inventors of the present invention have discovered that, unexpectedly, the use of the (-) enantiomer of limonene or 2-ethylhexanol, or a mixture of both, significantly improves the attractiveness of the ethanol / methanol mixture in the attraction of coffee borer Hypothenemus hampei. The coffee borer, Hypothenemus hampei (also known as coffee berry borer), is also called coffee berry borer or coffee berry borer. The coffee borer is a beetle of the family Curculionidae, feeding exclusively on coffee. It is present in almost all cultures of the world and almost all of its life cycle takes place in the fruits of the coffee, to the detriment of the seed. By "attractive kit for attracting the coffee borer" means a kit capable of significantly increasing the number of beetles in an attraction site of interest, compared to a control situation without said kit of attractants. The kit of attractants thus serves as "lure" or "bait" with respect to the coffee borer. Preferably, this kit of attractants is specific to the coffee borer, that is to say that the attractive effect of the kit of attractants is exerted preferentially on the coffee beetles, preferably predominantly on the bark beetles. coffee, and even more preferably only on coffee beetles.

"Attractive kit" means either a composition of attractants or several attractants used together. More particularly, the kit of attractants is adapted to distribute in the environment said attractants, preferably by diffusion. By ethanol / methanol mixture is meant a composition consisting of ethanol and methanol, preferably in the form of pure or essentially pure alcohols (that is to say more than 90, 95, 99 or 99, 5%).

By "about" is meant the value + or - 10%, preferably + or - 5% thereof. For example, about 100 means from 90 to 110, and preferably from 95 to 105. The inventors have demonstrated that the use of the (-) enantiomer of limonene and / or 2-ethylhexanol makes it possible to improve the attractiveness of said ethanol / methanol mixture towards the coffee borer.

Since limonene is a chiral molecule, it possesses two enantiomers, that is to say two forms of optical isomers which correspond to two structures, images of one another in a symmetry with respect to a plane, and of which the properties may differ. There are three kinds of nomenclature for differentiating enantiomers: R / S, D / L and +/-. The (-) enantiomer of limonene is also called L-limonene or S-limonene.

The (-) enantiomer of limonene is in an amount sufficient to significantly increase the attractiveness of the kit of attractants according to the invention. Preferably, the (-) limonene is in purified form. Thus, it is in pure form at more than 90, 95, 99 or 99.5%.

In a particular embodiment, the kit of attractants of the invention, containing the mixture ethanol / methanol, and (-) limonene and / or 2-ethylhexanol, further comprises a diluent. More particularly, said diluent is devoid of attractive activity towards said coffee borer. In particular, it can serve as a diffusion matrix for the attractants of the kit and thus influence the diffusion in the environment of said attractant. The diluent can be solid or liquid. The thinner can be a fatty substance without odor, waxes. It may for example be paraffin or a mineral oil. In a preferred embodiment, the diluent is paraffin. In a further embodiment, said attractant kit comprising the mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol further comprises one or more additional attractive molecule (s) (s). ). In a particular embodiment, the one or more additional attractive molecule (s) are volatile organic compounds. By "volatile organic compound" is meant any organic compound that can easily be in gaseous form in the atmosphere and able to spread more or less far from its place of emission. Preferably, said volatile organic compound is emitted by coffee cherries. Those skilled in the art will be able to choose the additional attractive molecules that do not interfere with the attractive effect of (-) limonene and / or 2-ethylhexanol.

In a preferred embodiment, the additional attractive molecule (s) are kairomones, in particular coffee grains or cherries. "Kairomone" means any substance produced by a transmitting living being (eg coffee beans) which, when released into the environment, triggers a response behavioral in another species (receptor), here the coffee borer, Hypothenemus hampei. The kairomones are beneficial for the receiver. Kairomones may be involved in the recognition of host plants by bark beetles.

In a particular embodiment, the additional attractive molecule (s) is (are) selected from the group consisting of: 2,4,6-cycloheptatrien-1-one, 2,6-bis (1), 1-dimethylethyl) -4- (1-oxopropyl) -phenol, 2-methoxy-3 - (1-methylpropyl) pyrazine, 2-methoxy-3- (2-methylpropyl) pyrazine, (2E) -tridecenyl acetate (5R, 7S) -conophthorine, (5S, 7S) -conophtorin, (E) -4,8-dimethylnona-1,3,7-triene, (E) -dendrolasine, [1,1'-bicyclopentyl] - 2-one, 1,1-dimethylcyclopropane, 1-butanol, 1-ethoxy-2-propanol, 1-hexanol, 1-hexen-3-ol, 1-methoxy-2-propanol, 1-methoxy-3-methylbenzene, 1-Methoxy-4-methylbenzene, 1-methylethylbenzoate, 1-methylethyl hexadecanoate, 1-nonanol, 1-nonene, 1-nonen-4-ol, 1-octen-3-ol, 1-pentanol, 1-phenoxypropan 2-ol, 1-propanol, 1-terpineol, 2- (E) -hexenal, 2- (E) -hexenol, 2- (E) -octenal, 2- (E) -pentenal, 2- (Z) - pentenol, 2,2-dimethyl-propanal, 2,3-butanediol, 2,6-di-tert-butylquinone, 2,6-dimethyl-7-octen-ol, 2-bu tenal, 2-butyl-1-octanol, 2-carene, 2-cyclohepten-1-one, 2-ethyl-1-hexyl acetate, 2-ethylhexanal, 2-heptanol, 2-heptanone, 2-heptyl acetate, 2 -indanol, 2-methoxy-3-methylpyrazine, 2-methylisobutyrate, 2-methyl-1,3-butadiene, 2-methyl-1-propanol, 2-methyl-2-butenal, 2-methyl-3-pentanol, 2-methylbutanol, 2-methyl-ethylpropanoate, 2-methylfuran, 2-methylpropanal, 2-methylpropyl acetate, 2-methoxy-3- (2-ethylpropyl) pyrazine, 2-nonanone, 2-pentanol, 2-pentanone, 2-pentyl acetate, 2-phenoxyethanol, 2-propanone, 3- (1-methylethyl) -2-methoxypyrazine, 3- (Z) -hexenol, 3,4-dimethylpentanol, 3,7-dimethyl-1,3,7 -octatriene, 3-ethyl-4-methylpentanol, 3-heptanone, 3-hexen-1-ol formate, 3-hydroxy-2-butanone, 3-isobutyl- (2-methylpropyl) -2-methoxypyrazine, 3-methyl- 1-butanal, 3-methyl-1-butanol, 3-methyl-1-butanol acetate, 3-methyl-2-butanone, 3-methyl-3-butenol, 3-methyl-butanol acetate, 3-methylfuran, 3- octanol, 3-octanone, 3- pentanone, 3- sec-butyl-2-methoxypyrazine, 4,7-dimethyl-4-octanol, 4,8-dimethyl-3,7-nonadien-2-ol, 4-hydroxy-4-methylpentan-2-one, 4-terpineol, 5-methyl-1-heptene, 6-ethenyldihydro-2,2,6-trimethyl-2H-pyran-3 (4H) -one, 6-methyl-5-hepten-2-one, acetaldehyde, acetic acid, hexanoic acid hexadecenoic acid, acora-3,5-diene, allo-aromandrene, anisole, benzaldehyde, benzyl alcohol, borneol, bourbonene, brocain (= 1,6-dioxaspiro [4,5] decane), butyl acetate, butyl butanoate , C2-benzene, C3-benzene, camphene, camphenilone, carboxilic acid, caryophyllene, caryophyllene oxide, chalcogran (= 2-ethyl-1,6-dioxaspiro [4.4] nonane), cis-calamenene, cis-linalool, cumene, curcumene, cymene, decanal, anethofuran (Case No. 74410-10-9), dimethyl-disulfide, dimethyl-propanedioic acid, dodecanal, elixene, ethanol, ethyl acetate, ethyl ester of the acid propanoic acid, ethyl isobutyrate, ethyl isovalerate, ethyl salicylate, ethylbenzene, ethylbenzene acetate, frontaline (= 1,5-dimethyl-6,8-dioxabicyclo [3.2.1] octane), geranial, geranylacetone, hexahydrofarnesyl acetone, hexanal hexanol, humulene, humulene epoxide, isobutyl acetate, isobutyl butyrate, isopentanol, isopentyl acetate, kaurene, limonene, limonene oxide, linalool, linalool oxide, longibornéol, longibornéol acetate , longifolene, m-cymene, menthol, menthone, mesitylene, methyl-2-hydroxy-6-methylbenzoate, methyl acetate, methyl hexanoate, methyl salicylate, methyl-2-hydroxybenzoate, methyl-3-ethyl-4-methylpentanoate , methylcyclohexane, m-methylanisole, myrcene, naphthalene, neral, nonanal, nonane, nonene, octanal, octane, octyl methanoate, oleyl alcohol, o-xylene, pI, 3,8-mentatrienol, p-1,3,8-menthatriene, para-xylene, p-cymene, pentanal, pentyl acetate, phenylethyl alcohol, prizizaene, propyl acetate, p-tert-butylbenzyl alcohol, pulegone, sabinene, sativene, styrene, thujene, thujone, thujopsene, toluene, trans-linalool oxide, trans-pyran linalool oxide , trans-P-ocimene, undecanal, undecane, α-bergamotene, α-cedrene, α-cham- lene, α-copaene, α-fenchocamphorone, α-isomenthone, α-longipinene, longipinene, α-muurolene, α-neocallitropsene, α- -phellandrene, α-pinene, α-terpinene, α-terpinolene, β-acoradiene, β-cedrene, β-elemene, β-gurjunene, β-himachalene, β-phellandrene, β-pinene, γ-butyrolactone, γ-Eudesmol , γ-muurolene, δ-cadinene and δ-germacrene.

More preferably, said kit of attractants comprising a mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol further comprises one or more additional attractive molecule (s) selected from group consisting of: 1-octen-3-ol, 2-heptanol, 2-heptanone, 2-indanol, 2-phenylethanol, 3-ethyl-4-methylpentanol, 6-methyl-5-hepten-2-one, acetate ethylbenzene, butyl acetate, isobutyl acetate, phenylethyl alcohol, benzaldehyde, brocain, caryophyllene, chalcogran, conophthorine, ethylbenzene, frontaline, humulene (also called α-caryophyllene), kaurene, linalool, longibornol, longipinene, methylcyclohexane, myrcene, nonane, limonene oxide, linalool oxide, ethyl salicylate, methyl salicylate, α-copaene, α-pinene and β-pinene.

Even more preferably, said kit of attractants comprising a mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol further comprises one or more additional attractive molecule (s) selected in the group of: α-pinene, β-pinene, caryophyllene, myrcene, 3-ethyl-4-methylpentanol, 2-heptanone, methyl salicylate, benzaldehyde and ethylbenzene.

Quantities, proportions and ratios

According to one embodiment, the kit of attractants according to the invention comprises a mass quantity of the ethanol / methanol mixture of between 0.5 g and 330 g, preferably between 0.5 g and 40 g, even more preferably between 40 g and 40 g. .

In particular, the kit of attractants according to the invention comprises a mass quantity of ethanol of between 0.1g and 300g, preferably between 0.5g and 80g, more preferably between 1g and 10g. In particular, the kit of attractants according to the invention comprises a mass quantity of methanol of between 0.1g and 300g, preferably between 0.5g and 250g, even more preferably between 3g and 30g.

In one embodiment, the kit of attractants according to the invention comprises a mass quantity of limonene of between 0.1g and 12g, preferably between 0.1g and 3g, more preferably between 0.2g and 1g.

In one embodiment, the kit of attractants according to the invention comprises a mass quantity of 2-ethyl-1-hexanol of between 0.01 g and 30 g, preferably between 0.1 g and 10 g, more preferably between 0.2g and 1g.

In a particular embodiment, the kit of attractants according to the invention comprises a mass quantity of additional attractive molecule (s) of between 0 and 12 g, preferably between 0 and 6 g, and even more preferred between 0 and 2g.

In another particular embodiment, the kit of attractants according to the invention comprises a mass quantity of diluent of between 0 and 100 g, preferably between 0.5 and 50 g, more preferably between 0.5 and 5 g. According to one embodiment, the kit of attractants according to the invention comprises a proportion of the ethanol / methanol mixture of between 15 and 99.98% by weight, preferably between 30 and 99.75%, even more preferably between 75 and 99%.

In a particular embodiment, the kit of attractants according to the invention comprises a proportion of (-) - limonene of between 0.02 and 50% by weight, preferably between 0.2 and 25% by weight. even more preferred between 2% and 10%>. In a particular embodiment, the kit of attractants according to the invention comprises a proportion of 2-ethyl-1-hexanol of between 0.01 and 50% by weight, preferably between 0.1 and 25% o, of even more preferably between 1% and 10%>.

In a particular embodiment, the kit of attractants according to the invention comprises a proportion of diluent of between 0 and 90% by weight, preferably between 0 and 50%, more preferably between 10 and 25%. .

In a particular embodiment, the kit of attractants according to the invention comprises a proportion of additional attractive molecule (s) of between 0 and 50% by weight, preferably between 0 and 25%. . The percentages are given here in relation to the total mass of the components of the kit (excluding diffuser).

According to one embodiment, the kit of attractants according to the invention comprises a volume ratio of ethanol / methanol of between 1: 10 and 10: 1, preferably between 1: 10 and 1: 1, even more Preferably, between 1: 3 and 1: 1. In a particular embodiment, the kit of attractants according to the invention comprises a volume ratio of (-) - limonene / ethanol / methanol mixture of between 5: 1 and 1: 500, preferably between 1: 1 and 1: 100, even more preferably between 1: 2 and 1: 15.

In a particular embodiment, the kit of attractants according to the invention comprises a volume ratio of 2-ethyl-1-hexanol / ethanol / methanol mixture of between 10: 1 and 1: 1000, preferably between 2: 1 and 1: 500, even more preferably between 1: 1 and 1:50.

In a particular embodiment, the kit of attractants according to the invention comprises a volume ratio of (-) - limonene and / or 2-ethylhexanol / additional attractive molecule (s) which is greater than 1: 2, for example 1: 1, 2: 1 or 10: 1. In a particular embodiment, the kit of attractants according to the invention comprises a ratio by volume of diluent / attractants which is less than 9: 1 (For example 8: 1, 7: 1, etc.), the attractants being the ethanol / methanol mixture, the (-) limonene and / or the 2-ethylhexanol and optionally the additional attractive molecule (s) (s).

The kit of attractants according to the invention may comprise the mixture ethanol / methanol, (-) limonene and / or 2-ethylhexanol, optionally the molecule (s) attractive (s) additional (s), optionally the diluent, according to the mass quantities (expressed in g) detailed above, and / or according to the proportions (expressed in%) detailed above, and / or according to the detailed ratios (in volume) above.

Distribution of attractants In a preferred embodiment, said kit of attractants comprises a device for the distribution of attractants in the environment. In a preferred embodiment, said attractive kit comprises one or more diffusers. The term "diffuser" means any device for the diffusion of the compounds of said kit of attractants. By "diffusion" is meant the propagation in the environment of the compounds of the kit of attractants according to the invention. Preferably, said diffusion is sufficient to allow the attraction of said coffee borer. More particularly, said diffusion corresponds to the evaporation of the compounds of the kit of attractants according to the invention, preferably the evaporation of the attractants of the kit of attractants, the attractants of the kit of attractants being the mixture ethanol / methanol, the (-) limonene and / or 2-ethylhexanol and optionally the additional attractive molecule (s). The attractants of the kit according to the invention are not necessarily mixed with each other in the diffusers, the aim being to create a mixture in the environment, preferably in air, after diffusion of each attractant together. According to a particular embodiment, the minimum duration of diffusion is 10 days, preferably the minimum duration of diffusion is 2 months. More particularly, different diffusion rates between the attractants of said kit may justify the use of several diffusers. Those skilled in the art are able to determine the content of the diffuser (s) according to said desired attraction diffusion rates.

Thus, the diffuser may comprise a single type of attractant, or any mixture of attractants, the attractants being the ethanol / methanol mixture, the (-) limonene and / or 2-ethylhexanol and optionally the molecule or molecules (s) additional attractive (s). Optionally, the diffuser may contain a diluent, which may serve as a diffusion matrix for the kit attractants, preferably a mineral oil or paraffin.

According to a first particular embodiment, said kit of attractants comprises:

- A diffuser comprising all the attractants used, ie the mixture ethanol / methanol, (-) limonene and / or 2-ethylhexanol, and optionally a diluent. In this context, the present invention also relates to an attractive composition of the coffee borer Hypothenemus hampei, characterized in that it comprises a mixture of ethanol / methanol, (-) limonene and / or 2-ethylhexanol and optionally a diluent. Preferably, said attractive composition comprises a mixture of ethanol / methanol, (-) limonene and / or 2-ethylhexanol and optionally a diluent with the characteristics of quantity and / or proportion and / or ratio of the components detailed for the attractive kit above.

According to a second particular embodiment, said kit of attractants comprises two diffusers: a first diffuser comprising the ethanol / methanol mixture, optionally the

2-ethylhexanol and / or a diluent.

a second diffuser comprising (-) limonene, optionally 2-ethylhexanol and / or a diluent.

According to this embodiment, the two diffusers comprise the attractants and optionally the diluent according to the quantities and / or proportions and / or ratios previously detailed.

According to another particular embodiment, the kit of attractants comprises two diffusers: a first diffuser comprising the mixture ethanol / methanol and optionally (-) limonene and / or a diluent,

a second diffuser comprising 2-ethylhexanol and optionally (-) limonene and / or a diluent.

According to this embodiment, the two diffusers comprise the attractants and optionally the diluent according to the quantities and / or proportions and / or ratios previously detailed. According to yet another particular embodiment, said kit of attractants comprises more than two diffusers: a first diffuser comprising the ethanol / methanol mixture and optionally a diluent.

a second diffuser comprising (-) limonene and optionally a diluent. a third diffuser comprising 2-ethylhexanol and optionally a diluent. In each of the above embodiments comprising two or more diffusers, it is also possible to provide that ethanol and methanol, instead of being included in the same diffuser, will be present in two adjacent diffusers (for example in the first and the second diffuser or in the second and the third diffuser) one of which contains methanol and the other ethanol.

Additional attractive molecules, chosen from the list mentioned above, may be added to one or more of the various aforementioned diffusers or be included in one or more additional diffuser (s). According to this embodiment, said diffusers comprise the attractants and optionally the diluent according to the quantities and / or proportions and / or ratios previously detailed.

In a particular embodiment, the attractants of the kit and / or the diluent as described above may be in solid or liquid form, preferably in liquid form.

In a particular embodiment, said diffuser is a container containing the attractant (s) and optionally the diluent, as described above. More particularly, said diffuser is a container containing a volume of attractants and optionally diluent of between 0.1 and 100 ml, preferably between 1 and 20 ml, preferably between 1 and 10 ml.

Device comprising the kit of attractants

The present invention also relates to a device comprising the kit of attractants as described above. The device comprises for example the one or more diffusers as described above intended to attract the bark beetle in an attraction site, and / or to modify its behavior or its orientation. The kit of attractants can for example be used in combination with pheromones and / or kairomones.

In a particular embodiment, the device comprising the kit of attractants according to the invention further comprises an element capable of killing and / or affecting the population of bark beetles. For example, it may include a wetting agent, an insecticide or a bark beetle pathogen such as Beauveria bassiana.

In another particular embodiment, the device comprising the kit of attractants according to the invention further comprises an element capable of trapping said borer. For example, the element capable of trapping said beaker is an element for sequestering said borer, to prevent it from evolving in its initial environment. In one embodiment In particular, the element capable of trapping said beaker is a container. Said container capable of trapping said beaker comprises openings, allowing entry of said beaker, the openings being perforations of variable diameters. More preferably, said container capable of trapping said beaker comprises openings whose shape allows said beaker to penetrate into said container, and prevents it from emerging. In a particular embodiment, the container further contains a capture substance. For example, said container may contain a capture liquid in which said borer drowns. In a preferred embodiment, said capture liquid is a detergent-type wetting agent solution such as TFD4. According to a preferred embodiment, the at least one diffuser as described above is placed in the container capable of trapping said beaker, preferably the at least one diffuser is fixed at mid-height in the container, preferably suspended at half height. in the container.

In a preferred embodiment, said element capable of trapping said beaker is a commercial trap BROCAP, UNITRAP, MULTIPHER III, LINDGREN. More particularly, said element is a trap UNITRAP or LINDGREN.

Methods and uses

According to another aspect, the present invention also relates to the use of the kit of attractants as described above, to attract the coffee borer Hypothenemus hampei. In another aspect, the present invention also relates to a method of attracting the coffee borer Hypothenemus hampei, wherein said kit of attractants is placed in or near a site of attraction.

Thus, the use of said attractant kit and the attraction method can be implemented for the manipulation of the behavior of the beetles. The kit of attractants can be used in particular to create confusion among coffee beetles, so that they are no longer able to locate their host, namely coffee beans or cherries. According to a particular embodiment, the arrangement of several kits of attractants, more particularly of several diffusers of said kit of attractants according to the invention, in and / or outside the coffee cultures can make it possible to disturb the recognition of the beaker against the natural attractants emitted by the coffee beans. The use of said attractant kit and the attraction process can also be implemented for biocontrol of bark beetle populations. More particularly, the kit of attractants can be used to kill the bark beetle or to infect it with a pathogen. In a preferred embodiment, the kit of attractants is included in a device further comprising a wetting agent or an insecticide for killing said attracted beetles, or a pathogen such as Beauveria bassiana to affect the population of bark beetles.

According to a preferred embodiment, the use for attracting the coffee borer and the method of attraction, are implemented for trapping said borer. According to a particular embodiment, said kit of attractants is included in a device further comprising an element capable of trapping said borer. The said device is placed near a coffee culture. Preferably, said device is placed in the coffee culture. Even more preferably, said device is placed directly in the coffee trees. Preferably, said device is suspended from the branches of the coffee trees, preferably at a height of about 1m or as high as possible when the coffee trees are smaller than 1m.

Trapping means sequestering said coffee borer, preventing it from evolving in its initial environment. More particularly, the attraction for trapping said bark beetle is implemented to preserve coffee crops. Said coffee crops can be from any locality in the world. More particularly, said coffee crops can be of any kind and variety of cultivated coffee.

Preferably, the trapping of the coffee borer is carried out at the beginning of the coffee berry formation. It has indeed been observed that mass trapping obtained with (-) limonene-based kits was all the more effective when fruit ripening was less advanced. It has also been demonstrated that the effectiveness of the kits can be maintained, even during fruit ripening, when 2-ethylhexanol is added to the kit, either in the same diffuser as (-) limonene, or in a separate diffuser.

EXAMPLES Example 1: Field trials in Brazil

Material and method : Trapping trials took place between January 8th and March 31st, 2016, during the fruit ripening phase, in a biodynamic Arabica coffee plantation in the municipality of Sào Joào da Boa Vista, in the state of Sào Paulo in Brazil, at an altitude of 800m. For each test, three plots hosted trials, each representing a randomized complete block.

The plantations of Coffea arabica var. tupi were made in the early 2000s by combining shade trees (Acacia mangium, Acacia auriculiformis, Gliricidia sepium, Leucaena leucocephala). The lines are contoured with varying plot spacing (0.6 to 0.75 x 3 to 3.6 meters), and shade trees are planted at regular intervals in the coffee rows. All plots benefit from important annual interviews: regular inter-row mowing, organic fertilization and weeding during fruit formation, foliar fertilization during grower growth and maturation. Yields, however, are moderate, ranging from 600 to 800 kg / ha / year. The traps (Unitrub fin model from Pherobank) were placed directly in the coffee trees, hanging from the branches at a height of 1.20 meters or as high as possible when the plants were smaller. The gap is about 20 meters between the traps.

The products used were provided by Sigma-Aldrich. It is :

Ethanol puriss. p.a.,> 99.8%, CAS RN: 64-17-5 Chromasolv® Methanol> 99.9%, CAS RN: 67-56-l

(S) - (-) - Limonene> 95%, FCC, FG, CAS RN: 5989-54-8

(R) - (+) - Limonene 97%, CAS RN: 5989-27-5

Paraffin oil, puriss., CAS no .: 8012-95-l The products were placed in glass vials of 2mL whose opening of the septum allows the free diffusion by evaporation. A first diffuser contained 0.5 ml of ethanol and 0.5 ml of methanol. A second diffuser optionally contained terpenoid diluted in paraffin (0.5mL of (-) - limonene or 0.5mL of (+) - limonene, 0.5mL of paraffin and, where appropriate, 0.1mL of (-) -a-pinene). The number of trapped coffee beetles was recorded daily, and the modalities tested were modified each week as a test. Each statement in each block represents a repetition. Between each test, the traps were washed with a 2% TFD4 solution and the diffusers were changed.

Results:

Study of the attractiveness of (-) - limonene associated with the ethanol / methanol mixture against coffee beetles.

Table 1. Number of coffee beetles trapped on the three plots tested (sum of the blocks), after one week of testing and according to 3 modalities: an empty diffuser (control 0); a diffuser comprising the ethanol / methanol (E + M) mixture; a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising (-) - limonene (E + M + (-) - limonene).

Table 1 shows that after a test (one week), the sum of Hypothenemus hampei trapped on the 3 plots tested is significantly increased by associating (-) - limonene with the ethanol / methanol mixture compared to ethanol / methanol mixture alone.

This result is confirmed by Table 2 which represents the total number of Hypothenemus hampei trapped for 3 weeks of testing (sum of 3 tests):

Table 2. Number of coffee beetles trapped on the three plots tested (sum of the blocks), over 3 weeks of testing (sum of 3 tests) and according to 2 modalities: a diffuser comprising the ethanol / methanol (E + M) mixture; a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising (-) - limonene (E + M + (-) - limonene). Table 2 confirms that the addition of (-) - limonene improves the attractiveness of the ethanol / methanol mixture. Indeed, the average number of Hypothenemus hampei trapped on the 3 plots at the end of the 3 tests is much higher when the limonene is added to the ethanol / methanol mixture, compared to the alcohols alone. Comparison of the (+) and (-) enantiomers of limonene.

The inventors have demonstrated that the attractiveness of the ethanol / methanol mixture is improved only with the (-) enantiomer of limonene. Indeed, the (+) enantiomer of limonene does not increase the number of trapped beetles when it is combined with the ethanol / methanol mixture, compared with the alcohols alone (see Table 3 below).

Table 3. Number of beetles trapped on the three plots tested (sum of the blocks), over two weeks of testing (sum of two tests), and according to 4 modalities: an empty diffuser (control 0); a diffuser comprising the ethanol / methanol (E + M) mixture; a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising (-) - limonene (E + M + (-) - limonene); a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising (+) - limonene (E + M + (+) - limonene).

Comparative test

A comparative test was carried out by substituting (-) limonene for another volatile constituent of coffee cherries, linalool, which is emitted in a significant proportion in the coffee breeze. The results of this test are summarized in Table 4 below.

Table 4. Number of beetles trapped on the three plots tested (sum of the blocks), over a test week and according to 3 modalities: an empty diffuser (control 0); a diffuser comprising the ethanol / methanol (E + M) mixture; a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising the natural linalool (E + M + linalool). Discussion

As expected, unattractive witnesses have almost never trapped bark beetles. The traps are therefore not attractive to bark beetles without attractant dispersers and the population density of bark beetles is small enough that catches are not due to chance.

On all tests, the addition of (-) - limonene to the ethanol-methanol mixture significantly increased the number of trap-trapped per day bark beetles compared to single alcohols. (+) - Limonene did not improve the catch compared to the alcohol mixture alone. In addition, the attractive effect of the ethanol / methanol mixture disappears almost in the presence of linalool.

Example 2: Field trials in Costa Rica

Material and method :

The above trials were completed in 2017 in Costa Rica in a context of larger bark beetle populations.

Trapping trials were conducted between February 21 and April 27, 2017 in CATIE's commercial coffee plantations (Centro Agronomico Tropical de Investigacion y Ensenanza) in Turrialba, Costa Rica. The number of repetitions per treatment varied from 6 to 9 according to the tests, the blocks having been constituted so as to be as homogeneous as possible.

The plantations of Coffea arabica var. caturra, located about 600 meters above sea level, were planted in 2010 in association with Erythrina poeppigiana as shade trees. The lines are contoured with a spacing of 80cm between plants and 2 meters between lines (6300 plants / ha). All the plots benefit from important annual interviews: weeding or chemical weeding of multi-annual inter-rows, chemical fertilization during fruit formation, fungicide treatments and possibly insecticides in case of bark beetle infestation. The yields are average, of the order of 1000 kg / ha / year of green grain coffee.

Brocap® traps were placed directly in the coffee trees, hanging from the branches at a height of about 1 meter and a distance of about 20 meters between the traps. The attractants tested are as follows:

Ethanol and methanol mixture: attractive marketed by Icafé (Instituto del coffee de Costa Rica), packaged in 15mL diffuser bottles, ethanol / methanol ratio 1: 3.

(S) - (-) - Limonene> 95% FG, Sigma-Aldrich, CAS RN: 5989-54-8 2-ethyl-1-hexanol (racemic), Sigma-Aldrich, CAS RN: 104-76- 7

Paraffin oil, purified, Sigma-Aldrich, CAS nbr: 8012-95-l

When all the products were diffused in mixtures, the alcohols and the complementary attracting compounds were arranged in vials of 4mL whose septum of PTFE was perforated with a hole of diameter 1/16 "(about 1,58mm) allowing the Free diffusion by evaporation When the alcohols were diffused separately, a vial of 2 or 4 mL perforated septum containing the terpenoid diluted in paraffin was associated with the commercial diffuser of alcohols.

The number of coffee beetles trapped was recorded daily. Each statement in each block represents a repetition. Between each test, the traps were washed with a 2% TFD4 solution and the diffusers were changed.

Results:

Confirmation of the interest of (-) - limonene associated with the ethanol / methanol mixture:

Table 5. Number of coffee beetles trapped on all the replicates (9 traps per treatment, 2 traps per trap), after 3 days of testing and according to 3 modalities: an empty diffuser (control 0); a diffuser comprising the commercial ethanol / methanol (E + M) attractive mixture; a diffuser comprising the ethanol / methanol mixture associated with a diffuser comprising (-) - limonene (E + M and (-) - limonene).

Table 5 demonstrates that the sum of the Hypothenemus hampei trapped on the 9 randomized repeats is markedly increased by associating (-) - limonene with the ethanol / methanol mixture relative to the ethanol / methanol alone mixture. Interest of 2-ethyl-1-hexanol associated with the ethanol / methanol mixture:

Table 6. Number of coffee beetles trapped on all the replicates (6 traps per treatment, 3 traps per trap), after 4 days of testing and according to 3 modalities: an empty diffuser (control 0); a diffuser comprising the commercial ethanol / methanol (E + M) attractive mixture; a diffuser comprising the ethanol / methanol mixture combined with a diffuser comprising 2-ethyl-1-hexanol (E + M and 2-ethyl-1-hexanol).

Table 6 demonstrates that the sum of trapped Hypothenemus hampei is markedly increased by combining 2-ethyl-1-hexanol with the ethanol / methanol mixture relative to the ethanol / methanol alone mixture. Ability to mix the components of the attractive kit:

Table 7. Number of coffee beetles trapped on all the replicates (6 traps per treatment, 3 traps per trap), after 4 days of testing and according to 6 modalities: an empty diffuser (control 0); a diffuser comprising the commercial ethanol / methanol (E + M) attractive mixture; a diffuser comprising the commercially attractive mixture ethanol / methanol and (-) -limonene (E + M + (-) - limonene); a diffuser comprising the commercially attractive ethanol / methanol mixture and 2-ethyl-1-hexanol (E + M + 2-ethyl-1-hexanol); a diffuser comprising the commercially attractive ethanol / methanol mixture, (-) - limonene and 2-ethyl-1-hexanol (E + M + (-) - limonene + 2-ethyl-1-hexanol).

Table 7 demonstrates that the number of entrapped Hypothenemus hampei is significantly increased by mixing (-) - limonene with the ethanol / methanol mixture relative to the ethanol / methanol alone mixture. The mixture of 2-ethyl-1-hexanol with alcohols allows also increase the trapping compared to the mixture ethanol / methanol. This increase is even sharper when the 2 compounds, (-) - limonene and 2-ethyl-1-hexanol, are added to the ethanol / methanol mixture.

Claims

1. Kit of attractants for attracting the coffee borer Hypothenemus hampei, characterized in that it comprises a mixture of ethanol / methanol and (-) limonene and / or 2-ethylhexanol.

2. kit of attractants according to claim 1, characterized in that it further comprises a diluent, preferably paraffin or a mineral oil.

3. kit of attractants according to claim 1 or 2, characterized in that it comprises one or more diffusers.

4. Kit of attractants according to claim 3, characterized in that it comprises a first diffuser comprising the ethanol / methanol mixture, a second diffuser comprising (-) limonene and optionally a diluent and optionally a third diffuser comprising the 2- ethylhexanol and optionally a diluent.

5. Kit of attractants according to claim 3, characterized in that it comprises two diffusers: a first diffuser comprising the ethanol / methanol mixture and optionally the

2-ethylhexanol and / or a diluent,

6. Kit of attractants according to claim 3, characterized in that it comprises two diffusers: a first diffuser comprising the ethanol / methanol mixture and optionally the

(-) limonene and / or a diluent,

7. Kit of attractants according to claim 3, characterized in that it comprises a diffuser comprising the mixture ethanol / methanol, (-) limonene and / or 2-ethylhexanol and optionally a diluent.

8. Device comprising the kit of attractants according to any one of claims 1-7, said device being adapted to trap and / or kill said borer.

9. Use of the kit of attractants according to any one of claims 1-7, or the device according to claim 8 for attracting the coffee borer Hypothenemus hampei, of preferably for trapping and / or killing said beaker or for modifying the behavior of said beaker.

10. A method of attracting the coffee borer Hypothenemus hampei, wherein the kit of attractants according to any one of claims 1-7 or the device according to claim 8 is placed in or near a site of attraction .