WO2021070060A1 - Insecticide device - Google Patents

Abstract

The invention relates to a biomimetic insecticide matrix in the form of a gel with characteristics capable of simultaneously guaranteeing the functionality of "lure and kill" for bloodsucking insects, in particular the tiger mosquito, and that of survival and growth of biocides to its internal active against bloodsucking insects. Said matrix comprises one or more biocompatible and biodegradable polymers, a pH between 4 and 8, salinity not higher than 3%, a quantity of free water higher than 0, preferably between 5-98% by weight and a viscosity comprised between 10^-3-10⁴ Pas. The matrix is able to maintain at least 30% of the initial weight within the fifteenth day of exposure to conditions of temperatures of 18-30°C and relative humidity of no less than 70%. The invention also relates to an insecticidal device which comprises said matrix.

Description

Insecticide Device

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Technical field

The present invention relates to an insect-killing device for the control of bloodsucking insects, in particular the device comprises matrix- based biopolymers with action "lure and kill" for indoor and outdoor applications, aimed at controlling the tiger mosquito population ( Aedes albopictus) and other hematophagous pests (ticks, lice, mites including dermanyssus gallinae, bedbugs, nematodes, annelids, fleas). The biopolymer-based matrix, also object of the invention, carries out its "lure and kill" action through the biomimetic attraction, the mechanical action, the activity and the self-dissemination of biocides proliferating within it.

The invention finds application in the "house-holding" field but also in the veterinary field, zootechnics, industrial pest control and in large- scale control campaigns, for example in parks, neighborhoods or cities. The invention also relates to ovitraps and other devices and supports which contain or use the matrix according to the invention. In particular, the invention increases the possibility of using such traps, devices and/or supports in the control of infesting insects as it is a solution with respect to the technical and cost-effectiveness problems described in greater detail in the state of the art.

The invention solves the technical problem of the absence of poisoned food baits or semiochemical attractants usually not suitable or not existing for bloodsucking insects (in particular the tiger mosquito), introducing the biomimetic approach, which also allows further advantages in environmental terms (eg effective use of biopesticides) and applications allowing the use of matrices and/or devices in scenarios where the use of pesticides or other chemicals is dangerous, not recommended or prohibited.

Prior art The growing incidence of resistance to chemical insecticides, the risks for human health and the environment and the regulations on pesticides have encouraged the search for alternative tools for the control of different species of insect vectors of diseases and in general of insect pests.

This problem has exploded in conjunction with the worsening of climate change and the globalization of the movement of men and goods that have led to the spread on a global scale of vectors and pests in places where they were not indigenous and in which they have found favorable conditions for their proliferation. We therefore found ourselves in the position of having to fight new vectors and/or pests with synthetic molecules that are less and less effective if used at concentrations that are safe for humans, animals and the environment. Another worsening condition is the conveyance of these substances through traditional methods such as the sprinkling/nebulization of aerosols. In fact, the latter, despite being cheap and easy to use, is not very effective in terms of targeting and requires repeated applications over time with increasing concentrations of insecticides, favoring the development of resistance to the active ingredients, increasing the risk of the presence of toxic molecules in the food chain, and the probability of involving insects that are not harmful and essential to the ecosystem such as primary pollinators (eg bees).

Therefore, solutions are sought that have costs comparable to those of the methods used up to now, but which at the same time increase the effectiveness and decrease the environmental impact of pest management actions. To obtain these results, on one hand, we try to increase the precision of the interventions and on the other hand we use biocides of natural origin (biopesticides), which on one hand have a lower environmental impact and are less likely to develop resistance phenomena on the part of insects, but on the other hand have higher costs and greater difficulties of application.

Excluding more systemic approaches, such as environmental interventions aimed at reducing possible proliferation habitats, the new "precision" methods are based, for example, on the semiochemical approach (use of sexual and/or food pheromones and/or natural/chemical flavorings) and/or poisoned baits (food approach). Both methods try to attract the target insect by conveying the insecticidal agent in order to attract the insect on traps, toxic surfaces or by making it ingest the insecticidal agent. However, such approaches can generate repellent reactions or be completely ignored by the insect [Tropical Biomedicine 30 (4): 691-698 (2013) Oviposition and olfaction responses of Aedes aegypti mosquitoes to insecticides, Canyon, DV] It is therefore necessary to think about new control methods, which exploit, for example, specific natural habits or behaviors characteristic of the target insect linked to essential activities of the life cycle (eg reproductive habits, search for water or shelters or places of deposition with specific features etc.). In other words, an approach that consists not in forcing the insect to interact with something it would never come into contact with naturally (eg a sheet smeared with glue), but in recreating and proposing an environment as close as possible to natural conditions ( in order to make the insect discard other artificial sites). In practice, imitating an environment, a surface, etc. in which the target insect would naturally perform a specific vital/biological function, making it at the same time lethal, by mechanical action and/or by the action of insecticides or natural bioactive substances. This approach can be defined as biomimetic, as it consists in replicating, or mimicking, on substrates/devices those physico-chemical, morphological, mechanical properties, etc. detected in the sites naturally chosen by a specific insect to carry out a specific activity, practically replicating a micro-habitat. Ideally, this micro habitat was at the same time also able to host biopesticides, allowing their survival and proliferation.

An approach that exploits the instinct of survival and conservation of the species, rather than proposing a solution that insects could naturally get used to and avoid (eg insecticide resistance). In order for this approach to be a valid alternative to the use of pesticides, it must be easily applicable (even on a large scale) but above all have an advantageous cost/effectiveness ratio. An exemplary case is that of the control of the tiger mosquito ( Aedes albopictus), vector of the arboviruses Zika, Dengue, Chikungunya and Yellow Fever. The tiger mosquito has adapted to live in both indoor and outdoor anthropized environments, it has its peaks of activity during the day and at night it prefers to take refuge in humid and difficult to access ravines which it often also uses as oviposition points. It is evident that - both for the impossibility of spraying insecticides in inhabited centers during the day and for the difficulty of targeting during periods of inactivity - specific control strategies and methods are needed and that possibly also involve biopesticides since it has been found resistance to different molecules ( Pyrethroid resistance in Aedes aegypti and Aedes albopictus: Important mosquito vectors of human diseases, Smith LB, Pesticide biochemistry and physiology, 2016 Oct. 133: 1 -12).

In the specific case of the tiger mosquito, among the possible alternatives there are control through ovitraps (with or without insecticide) and biological control through bioinsecticides such as microbial larvicides (eg. Bacillus thuringiensis var. Israelensis ) or symbionts (eg Wolbachia pipientis ), natural essential oils or through entomopathogenic fungi such as Beauveria bassiana (BB) and Metarhizium anisopliae.

An ovitrappola is a device used in the past almost exclusively for the collection of eggs for the purpose of scientific monitoring, then subsequently for the capture of pregnant females. It works based on the habit of the tiger mosquito females (natural habitat) to spawn (Thebiology o ec/esalbopictus, WA Hawley, Journal of the American Mosquito Control Association. Supplement 1988, vol 42-40):

• on vertical or otherwise inclined surfaces, placed in hidden areas and in shadow,

• in wetlands (or on which they perceive the presence or possible presence of water) and not directly into the water,

• in sites free of chemical which disincentive deposition or unpleasant general for the insect, • in environmental conditions of low salinity and pH tending to neutral.

In nature these places consist of rocky ravines, hollow plants, muddy areas on river banks, etc. In anthropized areas, similar conditions are found in hidden places such as pots and saucers, waste, abandoned tires, gutters, etc. The temperatures at which hatching takes place range from 20 to 40°C.

Therefore, to provide this attractive habitat, a trap in the simplest form consists of a container made of polymeric material filled with water with a vertically placed masonite rod inside. The masonite, when moistened, attracts the tiger mosquito to lay on it. The chopsticks are then removed and the eggs collected.

The transition from a monitoring tool to a control device led first to add synthetic insecticides to the water and then to new types of traps, considerably more complex, which avoid the use of insecticides but provide elements for the active capture of adults (eg . fans to suck them) or carbon dioxide cylinders and/or light signals to attract. This type of traps is intended for domestic use given the need for electrical power and the high costs. In attempts to use ovitraps in large-scale control campaigns (therefore with large numbers of traps to be placed on very large areas) the reference trap is the one used for monitoring with the addition of antilarval insecticides and any attractive substances.

These traps, when full of water, even if without the deposition substrate, act as deposition sites also for other species that spawn directly in water (eg. Anopheles, Culex, Aedes) therefore they are not specific for a single species of mosquito.

The problems relating to this last class of traps (defined as passive "lure and kill" traps as they do not use elements that do work to attract or capture, such as fans) are: a) the need to constantly top up the evaporated water and insecticide to function, b) keep them continuously monitored and therefore costs related to personnel, c) use of chemical insecticides with consequent environmental and interaction risk for animals and humans and non-target insects, d) need to recover traps when not active to prevent them from becoming new deposition foci if filled by rain (and therefore further costs for recovery and eventual disposal), e) low effectiveness due to competition with natural deposition sites in their vicinity The known solutions proposed go in the direction of creation of biodegradable traps to deal with problem d) but in any case based on the use of liquid water and insecticide (eg. Greenlid Biotrap).

Another solution is to make large standard traps to increase durability. In addition to management problems, increasing the duration to 10-15 days by using traps containing free water allows the larvae to reach the adult stage. It would therefore be necessary to add larvicidal products to the traps.

The effect of the laying substrate on the quantity of eggs laid in competition with natural sites is not evaluated and neither is the possibility of using substrates that retain water or absorb it from the environment (hydrophilic or hygrophilic) for periods of time compatible with the degradation of the trap or that imitate natural conditions (also due to the lack of quantitative information regarding the characteristic parameters of the substrates). The role of the substrate is significant as it influences the number of laid eggs ( Evaluation of attractants and egg- laying substrate preference for oviposition by Aedes albopictus (Diptera: Culicidae), U. Thavara, Journal of Vector Ecology, 2004, pp 66-72,). The standard substrate is a 20 c 2 cm masonite stick but for example BC Zeichner et al. used high-weight tissue paper soaked in an insecticide solution. Other examples of substrates are found in US patent 9237741 in which a hydrogel based on polyacrylamide (PAM) (therefore synthetic and non-biodegradable) is used as a deposition substrate to be inserted in a polyethylene container but also in US patent document 20130303574 in which a gel based on silica and talc is proposed which favors the contact transmission of an insect growth regulator or other synthetic insecticides to the insect.

In the documents mentioned above the trap:

- attracts the pregnant mosquito thanks to the presence of attractive substances and water (both in liquid state)

- kills the adult female by contact with a synthetic insecticide

- the deposition substrate, made of synthetic hydrogel based on polyacrylamide (PAM) or silica, is not biodegradable and is not biocompatible

- the substrate does not foresee and cannot act as a substrate for the growth of any biocide as it is not biocompatible.

Ultimately there are no studies that quantitatively characterize the factors that influence the choice of the deposition substrate (e.g. water content of the substrate, viscosity, composition, etc.), but operating conditions such as those of wet masonite can be characterized which has a free water content by weight between 30-50% calculated by differential scanning calorimetry (as described below), neutral pH and salinity lower than 3% (pH and salinity evaluated under standard conditions and depending on the absorbed solution).

In general, the possibility of hosting biocides replacing synthetic insecticides in the same deposition substrate has not been evaluated, an option that can only be achieved if there is biocompatibility between the material constituting the substrate and the biocide, a condition that is unlikely with synthetic polymers, and whether the substrate creates a suitable microenvironment for growth.

Take for example Beauveria bassiana (BB). This entomopathogenic fungus acts as a parasite of some types of insects, infecting them by contact (transmission of "lime" without the need for ingestion but through destruction of the cuticle by some enzymes and release of the toxin beauvericin), colonizing and killing them in a sufficiently long period of time long to allow the process of self-dissemination, that is the dispersion of the fungus by the parasite itself in other places infested or that will be infested by the parasite. Furthermore, the dead insect itself becomes a source of BB spores.

BB has assumed a key role in the management of numerous agricultural, veterinary and forest pests but also for the control of mosquito populations. In fact, it is effective on C. tarsalis, C. pipiens, A. aegypti, A. sierrensis, A. nigromaculis, and A. Albimanus ( Field and Laboratory Studies on the Pathogenicity of the Fungus Beauveria bassiana to Three Genera of Mosquitoes, Truman B . Clark et al. Journal of invertebrate Pathology, 1968, 11 , 1-7), but also against A. albopictus ( Characterization of Tolypocladium cylindrosporum- Hypocreales: Ophiocordy cipitaceae - and Its Impact Against Aedes aegypti and Aedes albopictus Eggs at Low Temperature, American Mosquito Control Association, Journal of the American Mosquito Control Association, 2017 Sep 33 (3): 184-192).

BB can be mass produced. Mass production usually involves using a mixture of cereals and water in a 1 : 1 ratio to which a suspension of BB spores is added. The ideal incubation conditions are 25°C and pH 4-8. It has been verified how the water content affects the growth of BB and in particular how the number of conidia produced with the same substrate grows with the increase in the amount of water ( Effect of Growing Media and Water Volume on Conidial Production of Beauveria bassiana and Metarhizium anisopliae, M. El Damir, Journal of Biological Sciences, 2006, 6 (2) -269-274). Beauveria bassiana is typically administered in one or more applications of conidia that are released into the air in dry or liquid formulations but also in aqueous or oily solutions, granules, pellets or floating formulations that exploit the possibility of the spores to multiply in an aquatic environment. The use of BB in biological control does not present any risk to human or animal health and its effectiveness against mosquitoes has been verified even when applied on substrates. For example, the spores of BB have been applied by spraying on earth, concrete and wood ( Storage and persistence of a candidate fungal biopesticide for use against adult malaria vectors, S. Blanford, Malaria Journal, 2012, 11 : 354) and mosquitoes (in this case Anopheles Stephens i and Anopheles gambiae) were exposed for 1 h to the treated substrate and then removed. After removal, survival was observed over the next 14 days and survival showed a strong dependence on the BB application substrate (confirming the link between application substrate and BB efficacy). Effectiveness and residual effect were also verified on plywood, bricks, plastic and glass surfaces {Persistence and efficacy of a Beauveria bassiana biopesticide against the housefly, Musca domestica, on typical structural substrates of poultry houses, Naworaj Acharya, Biocontrol Science and Technology , 2015, 697-715).

The use of BB is however strongly limited by the persistence times. In fact, a strong dependence of persistence on environmental conditions and on the substrate of application has emerged.

In particular, BB, if applied on earth, has a persistence comparable to that of a common pesticide (up to 4-5 months) while if applied on wood or concrete its persistence is reduced (from 1 week to 2-3 months). Differently if used in operating conditions (exposed to heat, UV radiation and drying of the substrate), the persistence drops to days.

In general, it has been shown that the encapsulation of the conidia of the fungi within polymeric matrices preserves their vitality and persistence. Procedures of this type are contained in the French patent No. 2,501 ,229, US Pat. No. 4,647,537, US Pat. No. 4,724,147, US Pat. No. 4,668,512. The inventions described relate to the encapsulation of pathogens inside matrices which involve gelling with the use of metal cations and the formation of a silica or silico-aluminate-based gel (Jung et al.) Or formulations based on alginate but in pellets or microspheres, therefore without any attractive functionality for the insect (Shigemitsu, Marois et al., Lewis et al.).

The characteristics of resistance and in particular of tolerance to thermal stresses are also strongly influenced by the composition of the growth medium of BB. For example, if carbohydrates are added to the preparation used for growth, BB can withstand temperatures of about 50°C ( Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana, S.-H. Ying, Applied microbiology, Sept. 2006, Vol. 43 pp 331 - 335).

In the direction of encapsulating the spores of BB and other biocides, patent US 5141744 moves towards the preparation of an insecticidal composition in the form of a hydrated macrogel that contains at least one species of entomopathogen and a compound capable of retaining hydration with the function of acting as a reserve of water for the entomopathogen itself. The purpose of the macrogel is on the one hand to maintain the vitality of the pathogenic organism by providing a useful means for positioning in insect infestation sites, and on the other hand to act as a bait for insects that are infected upon ingestion ("continuous insect consumable matrix").

For some insects, food baits are a valuable tool, but not for vectors of diseases such as mosquitoes, ticks, lice, mites, etc. as bloodsucking. Therefore the attraction with bait (as foreseen by the invention in the cited document) is not effective neither in terms of attractiveness, since it develops exclusively food-like attractiveness, nor in terms of infection since the biocide can act only if ingested. Furthermore, the gels described in US 5141744 are mixtures of matrices of natural origin such as oligopolysaccharides to which synthetic gels based on acrylic polymers, polyacrylamides and polyurethanes, which are not biodegradable, are added. Furthermore, said gels: a) contain chemical crosslinkers which in the case of the mosquito can act as a deterrent for the deposition as well as being toxic for the bioicides used, b) have no control over the amount of free water necessary for the survival and proliferation of the biocide and to the lure and kill action for example against mosquitoes (free water in the optimal configuration is between 70-80% by weight), c) they do not provide any indication on the rheological or yield stress properties that guarantee the possibility of use on vertical walls. The material proposed in US 5141744 acts exclusively as a protection to the pathogen and, as reported in the examples, does not show any growth of the biocide charge in the matrix rather, on the contrary, a reduction already after one week. Furthermore, the absence of nourishment does not favor the proliferation and survival of the pathogen for long periods.

There is also the disadvantage of dispersing a non-biodegradable synthetic polymer into the environment which, moreover, could also be ingested by non-target animals, children, insects, incurring limitations of use and therefore contradicting the need for innovation previously reported.

Similarly, US patent 5273749 describes a process for the coating of microbial pesticides but such as to be applied on leaves against plant weeds and subsequently ground and processed to be applied by spraying on plants or seeds. The invention has no lure and kill characteristics nor has it optimized characteristics to allow for a type of biomimetic attraction. In fact, the proposed formulation is designed to be sprayed on already infested leaves or soil or seeds and roots or for preventive purposes as would be done with any other insecticide. No characteristic of the gel that simulates an environment or condition that is naturally attractive to pests is sought or specified. The only role of the material is to provide a protective environment for the bioactive substance to act in the infested environment, without any entrapment functionality or the possibility of mechanical killing by suffocation of larvae or eggs. From no document of the known art does the idea or concept of a biomimetic approach to attract the insect emerge, that is to lure/attract it (lure functionality) by reproducing on a substrate or matrix of the conditions observed in nature and associated with a specific environment in which the target insect naturally carries out a certain activity typical of its behavior (eg mating, laying eggs, resting, sheltering, etc.). In other words, in no document known to the inventors are given to an insecticide matrix the same physical and chemical properties (with the same range of values) of a specific natural environment with the aim of attracting the insect (e.g. mimicking an environment with certain levels of humidity, pH, salinity on which the tiger mosquito goes to lay its eggs). Nor are there known matrices that use the biomimetic approach and at the same time are lethal for the attracted insect (kill functionality) through a mechanical action or the use of biopesticides. In literature there are not even matrices capable of performing the lure and kill functions and at the same time host biopesticides providing them with suitable conditions to preserve their vitality and favor their proliferation, with the aim of exercising the kill functionality without the use of chemical pesticides, which are often repellent in the attractive phase. Nor does it emerge the use of devices in which these conditions occur systematically and in a controlled and controllable manner.

All the devices describe a component of polymer and biocide but none of them contain indications on how to attract the insect effectively except with bait or through the use of substances with a semiochemical action (e.g. pheromones, aromas, etc.) with the risk, in addition, of the repellent effect of some biocides usually used to kill the insect. In other words, no document contains a technical solution to the problem of effectively attracting to substrates and/or lethal devices the types of insects for which effective bait or semiochemical substances are not defined. It follows that, although these technical solutions can be reported individually, no insecticidal matrix and/or device is described in the known art which presents the set of chemical-physical characteristics (with values within specific ranges) and composition that simultaneously allow the biomometric approach, the kill functionality and the growth and proliferation of a biopesticide. Finally, the matrix/device, configured in such a way as to perform all the functions indicated above, should also, for example, resist premature drying, be processable (eg spreadable, castable, injectable, etc.), stockable, have an adequate shelf life.

According to the inventors, certain characteristics of the matrix are functional to the characterization of the polymeric matrices and to the simultaneous success of the biomimetic effect (insect attraction) and to the incorporation of a biopesticide, to its survival and growth (matrix- biopesticide biocompatibility).

The conditions such as to carry out biomimicry, kill functionality and survival and growth of a biopesticide at the same time are not considered by the inventors to be within the reach of the expert in the field (or a team of experts) and therefore trivial, as they require the identification of a series of critical parameters (and their range of values) including salinity, viscosity and pH, percentage of water contained (free and total) composition (absence of insect repellent and/or toxic substances for the biopesticide), not known a priori in literature or not trivially coupled with each other as a simple sum. These parameters, if not present at the same time and/or if outside the identified ranges, result in a repellent effect against the target insect, as well as the absence of the kill function (when it occurs due to the mechanical action of the matrix) and of survival and proliferation of the biopesticide and transmission of biopesticides to target insects or the impossibility of practical application.

The identification of a composition that is not toxic (i.e. is biocompatible) to host biopesticides and a preparation protocol that does not damage the viability of the encapsulated biopesticides are considered out of the reach of an expert (or a team of experts) and therefore not trivial as they require the identification of polymers that at the same time possess biomimetic characteristics and have no toxic effect on the biopesticide and precise processing parameters such as temperature and preparation tools. The aspects of salinity, viscosity and pH, of composition and preparation are not taken into consideration by the known art because they are not considered functional since, in the inventions reported in the known art, they do not contribute to attracting the insect nor to guaranteeing biocompatibility of the biopesticide matrix, nor to kill it.

Only and only if all these aspects coexist, within the absolute and evaluated attractive ranges for the target insect and suitable for biocompatibility, then a device or a polymeric matrix allows to attract the insect without resorting to semiochemical substances and at the same time allow the survival of a biopesticide inside them that guarantees the killing of the target insect.

Therefore, considering the lack of a matrix and/or device that can simultaneously perform the biomimetic attraction, kill functionality and host and develop bioinsecticides, the problems posed by the known art remain unsolved to date. In particular, there is the problem of being able to attract a bloodsucking insect without the use of baits or substances with a semiochemical action (eg sexual, food or other pheromones); provide the insect with a reason to go to the polymeric matrix that is not the simple food motive (which obviously cannot work with a bloodsucking); provide the insect with a reason to go to the polymer matrix that is not due to chemical signals; to develop a polymeric matrix that performs all the aforementioned functions and that is at the same time lethal, biodegradable and biocompatible to host biopesticides.

SUMMARY OF THE INVENTION

The technical problem that the present invention aims to solve is that of effectively attracting on substrates and/or devices lethal for the adult and/or for its possible growth stages (eggs or larvae) of the types of bloodsucking insects for which are not deemed effective bait or semiochemicals.

Another technical problem that the invention aims to solve is that of providing an insecticide matrix which is at the same time biomimetic, lethal and capable of hosting a biopesticide, making it survive and proliferate. At the same time, with the proposed technical solution, the invention remedies the dangerous and ineffective use of synthetic insecticides (mainly due to the resistance developed by insects and the difficulties in targeting some species) using a "smart", inexpensive, extremely selective, eco-sustainable approach and with substances with a low risk profile that can be used without major restrictions. In particular, the invention increases the possibility of using devices such as traps, devices and/or supports in the control of pests in indoor and outdoor contexts and also in large-scale campaigns as it resolves some technical problems that compromised the cost-effectiveness ratio of the systems listed in the state of the art.

Therefore, the subject of the present invention is an insecticidal device consisting of a matrix with "lure and kill" operation, intended for the control in indoor and outdoor conditions of the population of bloodsucking pests (including also possible vectors of diseases) such as mosquitoes including the tiger mosquito, bedbugs, ticks, lice and mites like Dermanyssus gallinae, nematodes, annelids, papadaci, fleas, etc.

The matrix object of the invention performs simultaneously the "lure" functionality by exploiting the principle of biomimetic attraction and the "kill" functionality through mechanical action or with the aid of biocides, as well as performing the functionality of "survival and growth of the biocide" (i.e. to keep the biocide used for the target insect vital and effective and/or to proliferate) by exploiting the biocompatibility of the materials and the creation of a suitable microenvironment.

The invention is functional only when it has the characteristics suitable to simultaneously satisfy each functionality.

The matrix is prepared in the form of a gel in spreadable and/or pourable and/or moldable hydrated form or in dehydrated form. The gel is based on totally biodegradable, biocompatible, hydrophilic and hygrophilic biopolymers (ie able to absorb humidity and retain large quantities of aqueous solutions for prolonged times). By varying the composition and concentration of polymer, based on the type of polymer used, those parameters (e.g. pH, salinity, free water content, yield stress, etc.) that determine the properties necessary for operation, or for simultaneous satisfaction, of all features.

In practice, the matrix of the invention possesses the characteristics suitable for attracting a hematophagous insect by mimicking the characteristics of a natural habitat ideal for it for carrying out specific vital functions (including reproduction). For example, in the preferred composition, which has the tiger mosquito as its target insect, the matrix must mimic an egg-laying environment and for this reason it has a pH in the range 4-8, salinity in the range 0-3% (intended as a concentration of salts present in the solution and measured by conductivity measurements under standard conditions), yield stress threshold value of not less than 50Pa necessary for the gel to be spreadable and not pour on a vertical wall (measured with a rotational rheometer under standard conditions), a content of free water (ie not chemically bonded to the substrate) in the range 70-80% by weight (measured in standard conditions) and a viscosity of not less than 2 Pas.

At the same time, the matrix is capable of killing the insect and/or its eggs and/or larvae through the mechanical action of the matrix itself and/or thanks to the presence of a biopesticide. The mechanical action is carried out with the trapping and consequent suffocation of the eggs and larvae inside the polymeric matrix, an example is shown in figure 10.

If a natural biopesticide is used (eg entomopathogenic bacteria or fungi, etc.), it can survive and proliferate within the matrix (“survival and growth” functionality) up to lethal concentrations for the target insect and/or its eggs and/or larvae.

For example, in the preferred composition intended for the tiger mosquito, the conditions necessary for the "lure" functionality are suitable for the proliferation of the biopesticide Beauveria bassiana up to conidic concentrations of 10⁶-10⁷ conidia/mg of matrix, a quantity necessary for the "kill" functionality.

A further object of the invention is a matrix in hydrated, dehydratable and rehydratable form, which can be applied on a support, preferably biodegradable, able to be possibly inserted inside traps or other devices or containers functional to the creation of the attractive habitat or to the delivery of the matrix (such as in the case of containers in mosquito traps).

In the dehydrated form the matrix can be pre-shaped to be inserted into/on a support. The matrix will be pre- or subsequently rehydrated with a predetermined quantity of water until obtaining the same characteristics as the original hydrated matrix. Still another object of the invention are ovitraps and other devices and supports which contain or use the matrix according to the invention.

The matrix and the devices of the invention find application not only in the field of "house-holding", but also in the veterinary field, animal husbandry, industrial pest control and in large-scale control campaigns, for example in neighborhoods, parks or entire cities.

Further objects, objects and advantages will become apparent from the detailed description of the invention.

BRIEF DESCRIPTION OF THE FIGURES Figure 1. Cross section of a trap (2) containing the matrix (1) which attracts the tiger mosquito (3).

Figure 2. Viscosity values as the weight concentration of 2-hydroxyethyl cellulose (2-HEC) varies.

Figure 3. Values of free water in the matrix as the % wt (percentage by weight) of 2-hydroxyethylcellulose (2-HEC) varies.

Figure 4. Yield stress (Pa) values for different 2-HEC concentrations.

Figure 5. Water loss over time for a matrix made as in example 1 and tested in a climatic chamber inside a trap like the one in figure 1. Figure 6. T/C values as a function of the percentage by weight of 2- hydroxyethyl cellulose (% wt 2-HEC). T is the number of eggs deposited on the matrix compared to that deposited on control C. The curve shows the values of free water in the matrix as the % wt of HEC varies. Figure 7. Conidic composition-growth relationship (2-HEC with and without peptone). Evaluation of growth at the same concentration by weight of 2-HEC in matrices with and without peptone compared with two suspensions in distilled water with Beauveria bassiana (conidia and mycelium) respectively with and without the addition of peptone or other nutrients called respectively G and H. Figure 8. Comparative lethality test between matrices and suspensions previously subjected to growth test. The reference control against which mortality was assessed is distilled water (not shown in the figure). Figure 9. Comparative lethality test between matrices without Beauveria bassiana with weight concentrations of 2-FIEC between 2-30% prepared as in example 1. G and H: suspensions in distilled water of Beauveria bassiana (conidia and mycelium) respectively with and without the addition of peptone. K: distilled water with the addition of 1 % by weight of peptone. The reference control against which mortality was assessed is distilled water. The yield stress curves and the threshold yield stress are also shown.

Figure 10. Example of the mechanical action of the matrix and the effect of BB. A) Stereo microscope image of a larva trapped in the matrix. B) electron microscope image of an egg and the structures of the fungus grown on it.

Figure 11. The curves report the values over time of the number of conidia of Beauveria bassiana inside the matrix according to the composition reported in example 1 (A) and in example 2 (A freeze-dried). The stored A curve reports the values of the number of conidia over time present inside the lyophilized matrix and stored in a closed container and stored under standard conditions. Figure 12. Deterrent effect of the crosslinking process on matrix oviposition made according to Example 3.

Figure 13. Rheological and water content properties of the gels produced: A) yield stress value for different compositions as the concentration varies and threshold value to be overcome for applications of layers greater than 5 mm on vertical walls; B) viscosity variation as a function of the polymer concentration; C) weight variation of the gel (water loss) as a function of time (days) for different preparations; D) percentage of free water as the concentration varies for different preparations and reference values of masonite and wet cardboard as attractive substrates.

Figure 14. Role of pH in the deposition of the tiger mosquito expressed as T/C or ratio between the number of eggs laid on the sample and those laid on the control.

Figure 15. Role of salinity in the deposition of the tiger mosquito, expressed as T/C or the ratio between the number of eggs laid on the sample and those laid on the control. Figure 16. Deterrent effect on the oviposition of Aedes albopictus of lactic acid used as humectant and relationship with pH variation, expressed as T/C or ratio between the number of eggs laid on the sample and those laid on the control.

Figure 17. Deposition values of the tiger mosquito on matrices based on polymers alternative to HEC expressed as T/C or ratio between the number of eggs laid on the sample and those laid on the control.

Figure 18. Test of deposition against tiger mosquito in the field with different matrices in a period of 3 weeks expressed as T/C or ratio between the number of eggs laid on the sample and those laid on the control. Egg check carried out weekly.

Figure 19. Growth values of Beauveria bassiana and corrected mortality of gels containing and not containing Beauveria bassiana on tiger mosquito eggs. Figure 20. Behavioral evidence of Aedes albopictus. Comparative test in cage between different oviposition substrates in a period of 3 weeks expressed as T/C or ratio between the number of eggs laid on the sample and those laid on the control.

Figure 21. Effect of calcium chloride used as crosslinker in the deposition of the tiger mosquito expressed as T/C or ratio between the number of eggs laid on the sample and those deposited on the control. Figure 22. Viscosity variation over time during mixing (at shear rate 100 s ¹) of a formulation equivalent to a prior art formulation compared with the final viscosity value (at shear rate 100 s^-1) of the 16% HEC gel used as preferred formulation in example 1 (dotted line).

DETAILED DESCRIPTION OF THE INVENTION

The subject of the present invention is a matrix with activity and with a "lure and kill" function and devices with an insecticidal function containing said matrix, intended for the control in indoor and outdoor conditions of the population of hematophagous pests (including also possible vectors of diseases) such as mosquitoes including the tiger mosquito, bedbugs, ticks, lice, fleas, mites including Dermanyssus gallinae, nematodes, annelids, papadaci etc.

The matrix object of the invention performs simultaneously all the following functions. The "lure" functionality, carried out by exploiting the principle of biomimetic attraction; the functionality of "survival and growth of the biocide" (that is, to maintain vital and effective and/or to proliferate the biocide used for the target insect) which is carried out by exploiting the biocompatibility of the materials and the creation of an ideal microenvironment and functionality "Kill" which is carried out by mechanical action or with the aid of biocides.

The invention is functional only when it has the characteristics suitable to simultaneously satisfy each functionality.

The matrix is prepared in the form of a gel in a spreadable and/or pourable and/or moldable hydrated form or prepared in a dehydrated rehydration form. The gel is based on biodegradable, biocompatible, hydrophilic and hygrophilic biopolymers (capable of absorbing humidity and retaining large quantities of aqueous solutions for prolonged times).

By varying the composition and the concentration of polymer, based on the type of polymer used, the values of those parameters (e.g. pH, salinity, free water content, yield stress, etc.) that determine the characteristics necessary for operation, i.e. to the simultaneous satisfaction of all the functions specified above. In practice, the matrix of the invention has biomimetic characteristics, that is the characteristics suitable for attracting a hematophagous insect by mimicking a natural habitat ideal for it for carrying out specific vital functions (including reproduction). At the same time the matrix is able to kill the insect and/or its eggs and/or larvae through the mechanical action of the matrix itself, an activity linked to the entrapment and consequent suffocation of the eggs and larvae to the inside the material (an example is shown in figure 10) and/or thanks to the presence of a biocide. In particular, if a natural biocide is used (eg entomopathogenic bacteria or fungi, etc.), the latter can survive and proliferate within the matrix ("survival and growth" functionality) up to lethal concentrations for the target insect, before and after cold freeze drying operations.

A further object of the invention is a matrix in hydrated form which can be applied on a support, preferably biodegradable, able to be possibly inserted inside traps or other devices or containers functional to the creation of the attractive habitat or to the delivery of the matrix (for example as in the case of containers in mosquito traps).

In the dehydrated form the matrix can be pre-shaped to be inserted into/on a support. The matrix will then be rehydrated with a predetermined amount of water until the same characteristics of the desired hydrated matrix are obtained.

Still another object of the invention are ovitraps and other devices and supports which contain or use the matrix according to the invention. These inventions find application in the “house-holding” but also in the veterinary, zootechnical, industrial disinfestation fields and in large- scale control campaigns, for example in neighborhoods, parks or entire cities.

For the purposes of the description and within the scope of the present invention, the following definitions are given.

Functionality is defined as a specific task that the invention must perform. “Lure” functionality is defined as the ability to attract the target insect.

“Kill” functionality is defined as the ability to kill the target insect and/or its larvae and/or its eggs.

“Survival and growth” functionality is defined as the ability of the matrix of the invention to maintain vital and effective and/or to proliferate the active substance (biopesticide) used against the target insect.

The term "biomimetic attraction or biomimetic lure" means a method of carrying out the "lure" function not with traditional attractive substances, such as sexual aggregation pheromones and/or food attractants and/or other synthetic aromas with behavioral influence on insect, but providing the insect with a habitat with ideal conditions (mimicking those present in nature or using more attractive ones) for carrying out specific vital functions (including reproduction or development of offspring) typical of the target insect .

The term "kill by mechanical action" means a method of carrying out the "kill" function attributable to the action of the matrix alone without biocides or other active substances, carried out by trapping in the matrix of eggs that fail hatching or suffocation of any larvae (Figure 10). To carry out this action, the matrix does not need a biocide inside but is a capacity which, in the preferred formulation, is inherent in the material itself and linked to the viscosity of the material.

The term "kills (kill) through the action of biocides and/or active substances present in the matrix" means a method of carrying out the "kill" function attributable to the action of infection by biocides and/or other active substances present in the matrix.

By "biocidal product" we mean any substance or mixture (or generated from substances or mixtures) consisting of, containing and/or capable of generating one or more active ingredients, with the aim of destroying, eliminating and rendering harmless, preventing action or exercising other control effect on the target pests of the invention, by any means other than mere physical or mechanical action. Biocides also include “biopesticides”, consisting of naturally occurring or genetically modified microorganisms (such as entomopathogenic bacteria or fungi). The invention relates to this restricted class of biocides.

The biopesticides of natural origin that can be used in the invention include Gram-negative and Gram-positive bacteria, actinomycetes, fungi, protozoan yeasts, algae and their spores. Some examples are Bacillus subtilis subsp. Krictiensis, Pseudomonas pyrocinia, Pseudomonas fluorescence, Gliocladium wirens, Trichoderma reesei, Trichoderma harzianum, Trichoderma hamatum, Trichoderma viride and Streptomyces cacaoi subspecies asoensis; or microorganisms such as Bacillus thuringiensis. In particular, an example of a preferred biopesticide group that can be used within the present invention is constituted by entomopathogenic fungi belonging to four main groups: Oomycota (Lagenidium and Leptolegnia), Ascomycota (including Aspergillus, Beauveria bassiana, Metarhizium, Paecilomyces) and Zygomycota (including for example Conidiobolus, Entomophaga, Entomophthora, Erynia, Furia, Massospora, Neozygites, Pandora, Zoophthora). These substances are added to the starting solution to be gelled in variable quantities according to the type of biopesticide and subsequently proliferate in the matrix object of the invention. For example in the case of Beauveria bassiana used against the tiger mosquito the starting concentration is 10³-10⁴ conidia/mg of matrix with proliferation up to 10⁶-10⁷ conidia/mg of matrix.

By "bloodsucking insects" we mean that class of insects which have the feeding behavior of feeding on the blood of a vertebrate. In the present invention for hematophagous arthropods we mean those systematic groups of organisms belonging to arthropods which have as their alimentary behavior that of feeding on the blood of a vertebrate: by way of explanation but not exhaustive, different species belonging to the orders Ixodida (hard and soft ticks), Mesostigmata (parasitic mites), Anoplura (lice), Siphonaptera (fleas), Flemiptera Eteroptera (bedbugs), Diptera (mosquitoes, sand flies, horseflies, (Tabanidae), horse flies (Stomoxys calcitrans), etc.). In particular, as regards mosquitoes, reference is made to the genus Aedes, Culex, Anopheles. The term "biocompatible and biodegradable biomaterials" means natural hydrogels and among the most commonly used we can mention those belonging to the classes of: alginates, cellulose including for example hydroxyethyl cellulose, carboxymethylcellulose, pectins, polysaccharides, chitosans and agar. The biomaterials are added individually or in a mixture to the solution containing the biopesticide in concentrations that vary according to the biopolymer used. For example, the concentrations that allow the operation against the tiger mosquito using the biopolymer 2-hydroxyethylcellulose and Beauveria bassiana as biopesticide, are between 10-20% by weight. While using, for example, carboxymethylecellulose, the operating conditions are satisfied for concentrations between 2-10%.

Within the invention, a nutrient substance is intended as any element (or mixture of elements) which, added to the matrix, is useful for the life and metabolism of the living organisms present within it. Among the nutrients are considered carbohydrates, lipids and proteins, long chains of amino acids, minerals and vitamins in their simplest or complex forms and all their derivatives. For example, in the formulation used against the tiger mosquito, in the presence of Beauveria bassiana, 1% by weight of peptone (a water-soluble powder mixture of peptides and lipids) is added as a nutrient.

In the invention, a gel is intended as a biopolymer-based preparation consisting of a dispersed liquid incorporated in the solid phase, which includes or does not include biocides or nutrients that can be pourable, a spreadable or moldable paste, etc. obtained by adding a biopolymer to an aqueous solution or following rehydration of a previously dried matrix. In general, to ensure the operation of the invention, the parameters that characterize the gel, common to all functions (obtained experimentally or in accordance with the literature reported in the state of the art), are the following (measured under standard conditions): salinity not higher than 3%, pH 4-8, viscosity between 10³-10⁴ Pas and/or storage modulus and/or loss modulus values between 0-2MPa, yield stress values between 0-2000 Pa, in addition to a free water content greater than zero and preferably between 5-98% by weight (measured by differential scanning calorimetry). The biomaterials that make up the gel are added individually or in a mixture to the liquid in concentrations that vary according to the biopolymer used but such as to ensure that the gel respects the range of parameters reported. Within the invention, a humectant substance is intended as any element (or mixture of elements) with hygroscopic characteristics of natural origin which, added to the matrix, is useful for reducing dehydration and in general for maintaining the characteristics necessary for the performance of the previously defined functions. These include (as they are non-toxic and biocompatible as they are already used in the food industry) glycerin, propylene glycol, glyceryl triacetate sorbitol, xylitol and maltitol, polydextrose or natural extracts such as quillaia, compounds such as acid lactic or urea.

In the description of the invention, "quantity of free water" means that quantity of water/aqueous solution contained in the gel and not chemically bound to the biomaterial. It is expressed as a percentage of the weight of the gel and measured by differential scanning calorimetry with the method reported below.

In the description of the invention, salinity is intended as the concentration of salts dissolved in the gel which has been measured by electrical conductivity under standard conditions and is expressed as a percentage to be understood as grams of solute for every hundred grams of solution (mass fraction).

Therefore, for the purposes of the present invention by bio-matrix with lure and kill action through biomimetic attraction and mechanical action and of biopesticides proliferating inside it (called matrix for simplicity) we mean in the most general sense a preparation in the form of a gel which, possessing technical characteristics within the ranges shown, it allows to perform simultaneously the “lure”, “kill” and “survival and growth” functions defined above in the defined methods.

Method for preparing the matrix.

The matrix according to the invention is prepared with the following fundamental steps: In a temperature range of 15-60°C prepare an aqueous solution and add the polymer while continuing to mix (for 35-40 minutes for temperatures between 25-40°C at constant temperature) until complete gelling, which is highlighted by an increase in the viscosity of the solution (viscous solution such as oil for low viscosities or honey-like for higher viscosities), by the disappearance of the polymer powder and clarity. The complete gelling, regardless of the water/polymer ratios used, it is highlighted by an evident change in viscosity, by the disappearance of the polymer powder and by the passage from a cloudy solution to a gelatinous but clear one. At a temperature between 25-40°C the complete gelation takes place in about 40 minutes. The data was technically assessed by measuring the viscosity variation over time using a Malvern Kinexus rheometer (shear rate 100 s ¹). Complete gelling occurs when the viscosity remains constant. The aqueous solution can be previously prepared so as to contain a biocide and at least one nutrient for said biocide before adding the polymer and/or a wetting agent. Distilled water, mains water or water from non-potable collection tanks can be used. For example, in the case of the matrix intended for the control of the tiger mosquito, conidia and mycelium of Beauveria bassiana are added to the starting aqueous solution, made with distilled water, until a turbidity equal to 2 McFarland is obtained together with peptone (1% by weight) and finally the polymer (for example 2-FIEC, 16% by weight), mixing until gelation and in this case obtaining a gel with a pasty consistency. i.) As a possible additional step: pour into a mold and dry.

Preferably, if the matrix contains biocides, eliminate water, for example freeze-dry by freeze drying.

As indicated in the additional step, the matrix can be prepared in dehydrated form, for example with the freezedrying technique or another similar technique, known per se. In this embodiment, a matrix for example based on 2-hydroxyethylcellulose which includes and allows the entomopathogenic fungus Beauveria bassiana to proliferate inside it, is first prepared in the form of a gel, then it is poured or injected into a mold and subsequently subjected to the method freeze drying or dehydration by sublimation at low temperature and low pressure. With this method the entomopathogen remains viable but quiescent within the matrix and resumes its activity when the matrix is rehydrated. The matrix is easier to store and easy to use as it is already shaped and only needs to be wet to perform its function.

As previously reported, each function has parameters with defined operating ranges. The invention works when it satisfies the common operating conditions for the various functions.

To ensure the functionality of "biomimetic lure" the array must have the following characteristics:

• it contains a higher amount of free water at 0 and preferably between 5-98% by weight, in the case of the tiger mosquito in the preferred composition between 70-80% to be superior to the standard control,

• does not contain in the formulation of the substances which are repellents, for example for mosquitoes substances such as chlorine and in general the cations (eg. copper), flavors (eg. citronella and the like), chemical insecticides, acids, bases, etc.,

• it is processable in the form of a gel capable of covering a surface uniformly. In the case of the formulation against tiger mosquitoes, a support must be completely covered with a thickness of gel between 2-10 mm,

• it possesses, at the useful polymer concentrations suitable for the other functions, yield stress values such as to be able to be applied also on inclined walls. For example, in the case of the tiger mosquito, in order for a 5mm layer not to leak if placed on a vertical wall, the matrix must have yield stress values greater than or equal to 50 Pa (measured under standard conditions).

• pH between 4 and 8,

• a salinity of 3%, • maintains at least 5% free water by the fifteenth day of exposure under service conditions. For example the preparation for the control of the tiger mosquito, in order to remain attractive, must maintain at least 35% of free water on the fifteenth day of stay in the typical environmental conditions of the tiger mosquito deposition sites (18-30°C relative humidity greater than 70%),

• It contains biopolymers in concentrations such as to ensure the above characteristics,

• it contains wetting agents in concentrations such as to guarantee the previous characteristics. In the case of preparations for the tiger mosquito in concentrations ranging from 0-10% by weight.

To ensure the performance of the "biopesticide survival and growth" functionality, the matrix must possess the following characteristics, suitable for maintaining a habitat favorable to their survival, proliferation and infectious activity:

• contains at least 5% by weight of free water, a quantity that is also valid in the specific case of the preparation in which Beauveria bassiana is used,

• pH between 4 and 8,

• a salinity of 3%,

• it is prepared with processes that are not directly harmful to the vitality of the biopesticide or alter the conditions of the matrix necessary for the "survival and growth" functionality. For example, in the formulations in which the use of entomopathogenic fungi is foreseen, a temperature higher than 50°C cannot be used and neither sterilization methods of the BB-containing matrix that provide UV or extra heat or dehydration obtained with high temperature.

• it is composed of biopolymers and nutrients and/or wetting agents and/or other substances that are biocompatible, ie non-toxic towards the biopesticide, and in concentrations such as to respect the listed characteristics. The "kill" functionality (both mechanical and linked to the biocide) is guaranteed by the following characteristics of the matrix:

• performs a drying and/or trapping action on eggs and/or larvae and/or adults (mechanical action) regardless of the presence of the biopesticide, which is linked to the viscosity of the matrix. In the case of the preparation intended for the control of the tiger mosquito the viscosity value is not less than 2 Pas,

• allows the proliferation of the biopesticide up to lethal concentrations for the target insect, for example in the case of Beauveria bassiana it is necessary to reach concentrations of 10⁶- 10⁷ conidia/mg of matrix to be maintained for at least 10 days to be effective against eggs and tiger mosquito larvae,

• allows the self-dissemination action of the biopesticide if present, that is the possibility of the matrix containing the biopesticide (or of the biopesticide itself) to be transported to other places by the insects themselves that come into contact with the matrix. This occurs for viscosity values lower than 10²Pas (measured in standard conditions) and quantities of free water compatible with the “biopesticide survival and growth” functionality.

Furthermore, the matrix of the invention can be used inside devices or on supports, remedying the problems of lure and kill devices for the tiger mosquito (ovitraps) and other bloodsucking insects, if, in addition to satisfying the reported functions, It has the following characteristics:

• it is made of completely biodegradable polymers,

• it is formulated with a low risk profile elements to humans and animals and the environment,

• it has a durability, i.e. maintenance of the parameters necessary for functionality in a suitable range of values, of at least fifteen days in the classic conditions of the places where the tiger mosquito is deposited (temperatures between 18-30°C and humidity not less than 70%). The advantages associated to the matrix object of the present invention will be summarized in the following:

• it is multifunctional, that is, it carries out several functions at the same time: “biomimetic lure”, “survival and growth of the biopesticide” and “kill” not limiting itself to being only an attractive substrate or only growth or only lethal substrate.

• it targets a class of insects, the bloodsucking, which cannot be attracted with food baits or with semiochemical attractants but which responds positively to attraction with biommetic lure. The presence of attractants normally has both the role of attracting and "distracting" the insect from further chemical signals of substances that can be repellent (including the insecticides themselves). Assuming to eliminate the attractants from a formulation (for example food attractants such as sucrose, fructose syrup or apple extract and other non-food attractants such as sexual and/or aggregation pheromones) and considering the lack of attractiveness due to the rapid evaporation of water for extremely diluted gels (shown in example 11 and figure 22), there would be no type of attractive effect, making it effectively inactive and therefore ineffective.

• Absence of substances acting as possible deterrents for the attraction and therefore for the functioning and toxic for the environment of animals and humans including: insecticides, film forming materials for the creation of microspheres containing attractants, food attractants such as parts of other insects, UV protectors such as benzene derivatives or sulphonic acids and cerium or titanium oxide, humectants including polysiloxanes, and lactates (which as demonstrated in example 6 are repellent because they alter the pH), presence of acids and salts as thickeners (repellents in example 5 and 9). In fact, pH and salinity influence the functionality of biommetic lure as shown in Fig. 14 and 15 and where T/C values <1 , i.e. the ratio between the number of eggs laid on the control and on the matrix, indicate a direct correlation between salinity values or pH and deposition in the case of Aedes albopictus, highlighting that outside the ranges described as biomimetic there is no deposition on the gel. Therefore the absence of substances (including humectants) that could alter these parameters or be repellent is undoubtedly advantageous.

• manages to grow and develop the biopesticide up to concentrations such as to be lethal for the target species and allows the biopesticides to survive for long periods thanks to the advantage of its composition (which would die if dehydrated).

• it does not use synthetic pesticides but natural biopesticides or mechanical action. This guarantees: that the insect does not develop resistance, a low or no environmental impact and a potential absence of risk for humans and animals since no chemicals that may be toxic are used. Consequently, it guarantees a broader usage scenario. Let's consider a breeding context. One of the problems is the impossibility/dangerousness of using chemical insecticides to avoid contamination of products or animals. Gels containing insecticides, when used inside cages, for example of poultry for the control of Dermanyssus Gallinae, can lead to contamination of products or animals. If we eliminate the insecticide by the prior art gel, obtaining apparently similar compositions to the proposed matrix, they will be ineffective because they do not have the characteristics to perform an action insecticide differently from the gel described herein,

• can use biopesticides (such as entomopathogenic fungi such as Beauveria bassiana ) that do not need to act upon ingestion, solving the problem of poisoned baits, but also reducing environmental impact and selectivity,

• it is selective, as it allows to harm a specific harmful insect but not to other organisms that will not be attracted by the conditions mimicked by the matrix; • thanks to the hygrophilic and hydrophilic materials and form of a gel with slow release of moisture of which the matrix is composed and to its complete biodegradability,

• it can be used for extended periods without maintenance, without the need for water topping up, reducing service costs during disinfestations and eliminating disposal costs. Unlike the inventions of the prior art, which do not take into account the duration of the gel and the risk of its dehydration, as the quantity of water contained is only functional to the application by spraying on surfaces or as a means of dispersing attractants and/or insecticides and not to the attraction. That is, in the known devices the water is used as a simple diluent of the gel, that once sprayed and dries back in concentrated form, and not as an active part of the attraction,

• it is easy to prepare, as all preparations are at room temperature. The preparations are proposed in the prior art at high temperatures which would cause the death of any biopesticda present,

• it is easy to use and application as it can be coated by the user either by a coating line or dehydrated and rehydrated directly on the support to obtain the initial conditions,

• the preparations does not require the use of salts that promote the gelation such as sodium chloride or calcium which indeed have repellent effect as shown in Example 9

• the shelf life for the hydrated gel is at least about 1 year if stored in containers with adequate transpiration properties and for the de transpiration matrix it is almost infinite. Furthermore, the gel also protects the biopesticides from dehydration operations for storage purposes (eg freeze drying reported in example 2). The biopesticides therefore remains unchanged and active, guaranteeing an undeniable advantage also in terms of product shelf life.

• The polymers used are: - biocompatible because they allow the biopesticide to survive and proliferate for long periods of time,

- they are lure because they attract the hematophagous insect, but they are not necessarily bait for it, as it is not necessary for the hematophagous insect to ingest them to die, they only have the function of attracting the hematophagous insect,

- they are killers because they prevent the eggs from hatching and do not allow the survival of the hematophagous insect larvae, which die trapped in the polymer

- they are natural, renewable (vegetable source) and/or obtainable from scraps of other processes,

• The matrix, at least on mosquitoes and mites, is effective even without biopesticide, by mechanical action alone against bloodsucking insects, it was in fact unexpectedly found that it is the matrix that by mechanical action prevents the hatching of the eggs of bloodsucking insects or, when eggs hatch, traps the insect or the larva, which die because they do not find within the matrix neither the mobility nor the oxygen sufficient for their survival

• The matrix, like any other element of the formulation, is completely biodegradable and compostable and does not require further disposal operations, as it can be discharged among organic waste, reducing the environmental impact in terms of pollution, waste production and ease of disposal of the devices produced. It increases the safety and the possibility of use in indoor scenarios

• Besides being biomimetically attractive, the matrix also has a lethal effect on adult individuals even without the use of biocides and/or biopesticides but by simple mechanical action active on contact due to the composition of the gel, as demonstrated in the test subsequently reported in example 4. This effect allows to obtain a series of advantages linked to the absence of chemical biocides such as reduced ecological impact, possibility of use in indoor contexts including domestic ones in the presence of people and animals, elimination of possible resistance to classes of molecules by insects. Advantages are also obtained in terms of attractiveness of the substrate, which can be compromised by the repellent effect of some insecticides. In the proposed prior art, all insecticidal formulations present synthetic molecules and/or formulations for which ingestion by the insect is required.

• The matrix can withstand cycles of dehydration and rehydration without losing its ability to lure and kill ensuring an advantage in terms of storage and conservation but also to use prolonging the possibility of use,

• The biopesticide (such as beauveria) can be added to the matrix for the sole purpose of infecting the adult that settles on the matrix itself, but the action on the eggs laid and on the larvae is borne by the matrix, which prevents the eggs from hatching or the larvae from surviving killing them by suffocation due to the reduced presence of oxygen and the inability to move to emerge to look for it or to feed due to the viscosity of the matrix

• The various conditions of pH, salinity etc ... can certainly be found in a natural environment, such as a bowl or a fountain full of water, so they attract the insect, but these same characteristics, inserted in a polymer like those of invention, which are also characterized by a certain viscosity, modulus and yield stress, allow to kill the insect's progeny. And these characteristics allow us to say that the polymer is both attractive and an insecticide in itself, as its nature allows the insect to consider it a safe environment to lay eggs (or to perform other vital functions) and safe for the offspring, while instead it is the matrix that attracts the insect but kills its offspring.

• The viscosity values (which increase proportionally with the increase in the polymer/water ratio as described in figure 13 Panel b) allow the matrix of the invention to be used on different surfaces as the gel can be spread with the appropriate thickness on horizontal surfaces, inclined and vertical, or to fill containers in cases of lower viscosities. The matrix thus also exerts a lethal mechanical action (as indicated above on eggs and larvae) and is attractive (because the water contained in the gel does not evaporate quickly) and allows the survival and growth of the bioinsecticide. From the reported tests it appears that these conditions are attractive, for example, for the deposition of the tiger mosquito. In fact, gels with such viscosities are effective in blocking the hatching of eggs and in blocking the larvae. As an example, the yield stress threshold values are reported in order to use a 3 mm gel layer by spreading on a vertical surface (Fig. 13 panel a).

• In the matrices of the invention, the presence of crosslinkers such as strong bases (NaOFI), poly-ethylene glycol-diglycidyl ether

(PEGDE), or acid residues necessary for the neutralization of the alkaline conditions required for the crosslinking is not necessary. The presence of these components would compromise the attractiveness and deposition of eggs, as exemplified in example 3 and in Fig. 21 (deposition by Aedes albopictus on cross-linked alginate)

• The use of slow-release water gels and the absence of liquid water, used for example in large quantities in the known art to favor spraying with nozzles by lowering the viscosity, further reduces the environmental impact.

A humectant can be added to further slow down the evaporation phenomena for the gel when applied.

According to a preferred embodiment, the matrix is intended for the control of the tiger mosquito ( Aedes albopictus) and is a preparation based on 2-hydroxyethylcellulose (2 -FIEC) which includes and allows the entomopathogenic fungus Beauveria bassiana to proliferate. It is made in the form of a gel with characteristics such as to simultaneously perform the functions of "biomimetic lure", be spreadable and act both as an egg-laying substrate, attracting the tiger mosquito to lay, and as a lethal matrix through the mechanical action of the polymer and of the Beauveria bassiana proliferating in the matrix. Furthermore, the mosquito contaminated by the entomopathogenic fungus will guarantee self-dissemination in other spawning sites. The use of the matrix inside a trap is shown in figure 1 which shows a cross section of a set up of use of a trap with the matrix inside which is placed in direct contact with the vertical walls.

Other preferred polymers are Sodium Alginate (SA) and Carboxymethylcellulose (CMC) for the creation of matrices with formulations containing and not containing biopesticides such as Beauveria bassiana. With these polymers it is possible to obtain gels with the characteristics considered essential for the biomimetic approach towards the listed insects and in particular it is possible to obtain gels with chemical-physical properties (e.g. pH, salinity, viscosity, etc.) within the attractive ranges for the tiger mosquito in the same way as the 2-HEC. In fact, with gel formulations obtained starting from SA and CMC it is possible to obtain the same biomimetic attraction effect, recorded with 2-HEC and shown in figures 17 and 18 (field tests) as well as the same biocompatibility towards Beauveria bassiana as shown in the figure 19.

The technical problem that the present invention aims to solve is that of effectively attracting on substrates and/or devices that are lethal for the adult and/or for the possible stages (eggs or larvae) of the bloodsucking types of insects for which no effective baits or semiochemicals are defined. At the same time, there is no document in the literature that reports an insecticide matrix that is biomimicry, lethal and at the same time able to host making it survive and proliferate a biopesticide. At the same time, with the proposed technical solution, the invention remedies the dangerous and ineffective use of synthetic insecticides (mainly due to the resistance developed by insects and the difficulties in targeting some species) using a "smart" approach, inexpensive, extremely selective, eco-sustainable and with substances with a low risk profile that can be used without major restrictions. In particular, the invention increases the possibility of using devices such as traps, devices and/or supports in the control of pests in indoor and outdoor contexts and also in large-scale campaigns as it resolves some technical problems that compromised the cost-effectiveness ratio of the systems listed in the state of the art. The following examples have the purpose of illustrating the present invention, without limiting its scope of protection. It will be apparent to those skilled in the art that some modifications can be made to the present invention without departing from the spirit or scope of the invention as set forth herein.

EXAMPLES

Example 1: preparation of a matrix based on 2-hydroxyethylcellulose (2-HEC) peptone and Beauveria bassiana (BB), intended for the control of the tiger mosquito (Aedes albopictus). According to a preferred embodiment, the matrix is intended for the control of the tiger mosquito ( Aedes albopictus) and has the following composition: distilled water, 20-300 g/liter of 2-hydroxyethylcellulose powder (2-HEC, Sigma Aldrich) molecular weight medium 90000, 10g/liter of peptone for mycology (Sigma-Aldrich), 10 mg/liter of spores and mycelium of Beauveria bassiana strain CD 1123.

In the preferred embodiment the procedure for the preparation to obtain about 120 g of matrix at 16 % by weight of 2-HEC provides (starting from the conditions of sterility of the materials and instruments): a) prepare 100 ml of sterile distilled water, b) add 1 g of peptone and mix at room temperature until completely dissolved, c) add 1 g of conidia and mycelium of Beauveria bassiana strain CD 1123 (corresponding to a turbidity of 2McFarland and an initial concentration of about 10³conidia/ml of suspension), d) mix at room temperature for 10 minutes, e) add 16 g of 2-HEC and mix at room temperature for 40 minutes until a solution with a pasty consistency is obtained (estimated time for the complete gelling of the 2-HEC which occurs through a hydration reaction. The time was verified by analyzing the viscosity variation over time under standard conditions). The same preparation was performed in the experiments in the BB-free or peptone-free forms. In the description of the experiments, these alternative compositions will be referred to respectively as "prepared according to example 1 but without BB" and/or "prepared according to example 1 but without peptone".

These preparations all have pH values between 6-7 and salinity values below 3% under standard conditions (measured respectively with a Mettler Toledo model pH8 benchtop pH meter and a Bormac model COND 70+ portable conductivity meter).

Tests carried out

Tests were carried out which aim to demonstrate that the matrix prepared as shown in example 1 works against the tiger mosquito (ie it simultaneously satisfies the functions of "biomimetic lure", "survival and growth of the biocide" and "kill") and that the values of the parameters necessary for the operation of each functionality are satisfied and within the range of the more general ones reported in the description of the invention.

To evaluate the "lure" functionality, an oviposition test was carried out on the matrices made as in example 1 when the 2-HEC concentration changed. To evaluate the "survival and growth of the biocide" functionality, biocompatibility and conidic growth tests were carried out on matrices prepared as in example 1 (using the concentrations that had proved to be the most captivating in the deposition test). The growth test was combined with the lethality test on tiger mosquito ( Aedes albopictus) eggs to evaluate the “kill” functionality. The test was carried out at the end of the growth period of the Beauveria bassiana inside the matrix prepared according to example 1 . Tests were carried out to evaluate viscosity, percentage of free water, yield stress. The values obtained guarantee on the one hand that the preferred preparation reported in example 1 falls within the parameters that define the invention and furthermore that the values relating to the preferred preparation are really effective against the tiger mosquito.

Materials and methods and results Measurement of the viscosity of the matrix means

The matrix prepared as reported in Example 1 with concentrations by weight from 2 to 30% of 2-HEC but without Beauveria bassiana was subjected to a test to evaluate its viscosity. The test is of the "single shear rate" type performed with a Malvern Kinexus pro rotational rheometer in flat-flat configuration. The test was performed with constant shear rate equal to 50s ¹ for 5 minutes at 30°C and atmospheric pressure on each concentration of 2-HEC tested. The viscosity values varying in weight concentration of 2-HEC (2-30%) at 30°C and atmospheric pressure are shown in figure 2. The results show that the functionalities that require a viscosity value of not less than 2 Pas are guaranteed by weight concentrations of 2-HEC not lower than 4%.

Measurement of the quantity of free water present in the matrix By free water we mean water that is not chemically bonded to the material and that therefore can freeze and melt at the characteristic temperatures of water and is the first to evaporate in environmental conditions. The test was performed with differential scanning calorimetry (DSC) with the Q2000 Series instrument by TA Instruments, on matrices with 2-HEC concentrations in the range 2-30% by weight prepared as in example 1 but without Beauveria bassiana. The test (performed in air) foresees a cooling ramp at 5°C/minute down to -20°C, an isotherm at -20°C for 10 minutes and a heating ramp at 5°C/minute up to 50°C (atmospheric pressure). From the analysis of the endothermic melting peaks it is possible to obtain the quantity of free water inside the material. In particular, knowing the energy necessary for the fusion of a known quantity of water and peptone, it is possible to obtain the quantity of free water by analyzing the energy associated with the endothermic peak (area under the curve measured with the specific functionality of the software associated with the tool). The same procedure was carried out on a masonite sample immersed in water for 48 h. Masonite was used as the lower limit of free water necessary for the deposition contained on a standard substrate which is certainly attractive. The results are reported in figure 3 and show, as the concentration by weight of 2-HEC in the matrix prepared according to example 1 varies, the values of the free water contained in percentage with respect to the weight of the sample. The same figure also shows the value of free water contained in a sample of wet masonite. It is noted that as the concentration increases, the free water content decreases. For the matrix with 16% concentration the free water is 78% of the total weight of the sample.

Yield test The yield test is used to measure the maximum yield stress value or the tangential stress value at which a fluid begins to flow. It was performed with a Malvern Kinexus Pro rotational rheometer in the flat- flat configuration at a temperature of 30°C and atmospheric pressure. The threshold value was calculated as the tangential stress value to which a 5mm layer of matrix is subjected if placed on a vertical wall, when subjected exclusively to its own weight. If the yield stress peak value is lower than the threshold value then the matrix placed on the vertical wall will leak.

The tests were carried out as the concentration of 2-HEC by weight on matrices prepared according to example 1 but without Beauveria bassiana to avoid bad readings by the instrument. The absence or presence of BB does not involve significant changes in the yield test values.

The curve in figure 4 shows the yield stress values as the concentration by weight of 2-HEC of the matrix prepared as in example 1 increases. It is noted that the threshold value is exceeded starting from concentrations higher than 8% by weight of 2-HEC calculated so that a 5mm thick layer does not drip if spread on the vertical wall. Therefore, the 16% matrix made as in example 1 complies with the requirements of the various functions.

Weight loss over time test

The weight loss test was performed in a Binder model KBF 115 climatic chamber at 25°C and 70% relative humidity. In the test, the weight of a sample subjected to controlled environmental conditions is monitored daily. In the case of the matrix, the weight variation is associated with the evaporation of free water. The matrix is attractive if after 15 days it has at least a quantity of free water equal to that of wet masonite (substrate classically used for deposition) which is between 30 and 50% by weight (in this specific case it is equal to 35% ).

The matrix prepared as in example 1 was spread on the walls of a trap like the one shown in figure 1. The trap and matrix system was weighed daily and the net weight of the matrix obtained. Figure 5 shows the values of the normalized weights of the matrix (on the initial weight) with respect to time. We can see the weight variation or the loss of free water over time (the weight lost is associated with the evaporation of free water). The test shows how, even after 24 days, there is a weight of approximately 30% of the initial weight of the matrix which, starting from a matrix prepared as in example 1 which has 80% of free water, means having a quantity of free water equal to about 34% of the total weight of the matrix and therefore sufficient to be attractive since it is comparable to the limit condition established by the masonite.

After 15 days the matrix retains about 54% of the initial weight corresponding to a percentage of free water equal to about 60%, which is higher than the limit quantity defined by the humid masonite.

Furthermore, figure 5 shows the variation curve of the weight over time of the wet masonite (kept submerged until equilibrium is reached, i.e. the impossibility of absorbing further water). It is noted that the weight loss due to water evaporation is much faster in the masonite than in the gel.

Oviposition test for evaluation of biomimetic "lure" functionality.

One of the purposes of the matrix is to mimic a natural habitat of the tiger mosquito. The attractiveness of the matrix prepared as in example 1 is obtained from the test as the 2-FIEC concentration varies and the correlation between the characteristics of the substrate and the attractiveness is verified. For the purposes of the test, a colony of tiger mosquitoes was reared in an insectarium placed at 26°C and 70% relative humidity and with a ratio of hours of light: hours of darkness 14:10. Mosquitoes had a blood meal 48 hours before laying (time needed for the eggs to mature). At the end of the maturation time, 30 pregnant females were kept in the cage. Inside the cage were placed 2 plastic containers with 30 ml of distilled water. The control (absorbent paper, C) was placed on the internal walls of the first container. The matrix prepared as in example 1 , but withoutwas manually spread on the walls of the second container Beauveria bassiana (T). After 24 hours the containers were recovered and the eggs counted. The test was repeated 3 times for each 2-HEC concentration tested.

The values of the number of eggs laid on the matrix as the concentration of 2-HEC vary in the histogram of figure 6. The reported T/C value represents the ratio between the number of eggs laid on the tested matrix T and the number of eggs laid on the standard control C (damp absorbent paper). When the T/C value is greater than 1 then the matrix is more attractive than the control. The curve of figure 6 instead shows the value of the free water contained in the matrix used and the link between the free water parameter and the number of eggs laid on the matrix is highlighted with the definition of an optimal range of 2-HEC concentration. The results show that the best deposition results are obtained between 70% and 85% of free water or for concentrations between 8% and 20% by weight of 2-HEC and in particular the composition with 16% of 2-HEC (78% free water) is the most attractive.

However, the matrix has yield stress values that make it possible to use it on a vertical wall only starting from 10% of 2-HEC as the yield stress value is higher than the threshold value shown in figure 4.

Biocompatibility test and lethality (evaluation of the "survival and growth" and "kill" functions)

Since the matrix prepared as shown in example 1 must perform the "survival and growth of the biopesticide" function but also the "kill" function, two tests were performed in series: a biocompatibility and growth test of BB in the matrix and an efficacy test on tiger mosquito eggs (carried out using the matrices following the BB growth test). The tests were performed on the matrix prepared as in example 1 at the concentration of 2-HEC which recorded the best result in terms of attractiveness in the oviposition test.

Biocompatibility and growth test of Beauveria bassiana in matrices

The test evaluates, through conidic count, the impact on the vitality and growth of the biopesticide by the biopolymer used, the process of making the matrix and the microenvironment created by the matrix itself. The conidic count is used as the conidia are the infecting part of the entomopathogenic fungus therefore: the higher the conidial concentration the higher the infectious power.

The test was carried out on the matrix with the concentration of 2-HEC which obtained the best result in terms of oviposition (16% of 2-HEC by weight) prepared as in example 1 and containing Beauveria bassiana. A 16% preparation of 2-HEC containing 1% by weight of peptone and one without peptone were tested. The two types were compared with two suspensions based on distilled water (called G and H respectively) containing conidia and mycelium of Beauveria bassiana at the same initial concentration present in the matrix (shown in example 1). G contains 1% by weight peptone while H lacks it.

In this way, the test also evaluates the effect, with the same concentration of 2-HEC, of the presence of the peptone as a nutrient for Beauveria bassiana.

All matrices and control solutions are stored in sterile glass containers in a Binder model KBF 115 environmental chamber at 28°C and 80% relative humidity for 24 days (period comparable to the expected service time for the matrix). During this period a conidic count was carried out every 4 days by seeding on a plate for mycological growth based on potato dextrose agar (PDA) both on the matrices and on the suspensions. Conidic counting is performed with dilutions of the matrix in distilled water in the range ⁴10-10¹. The “number of conidia-days” curves shown in figure 7 compare the number of conidia of the entomopathogen Beauveria bassiana in the different growth media. The results show that, with the same concentration of 2-HEC, in the matrices without peptone the conidic charge is constantly lower. Furthermore, it can be noted that for suspensions, with and without peptone, the conidic charge is already lower than that of the matrices starting from day 8. Therefore the matrix prepared as in example 1 not only satisfies the "survival and growth" functionality but amplifies the growth of BB compared to a standard aqueous suspension.

Effectiveness of the matrix on tiger mosquito eggs ( Aedes albopictus) for different egg-matrix contact times

The same matrices used in the growth experiment and stored for 24 days under controlled conditions comparable to those of use of the matrix in the field (temperatures 18 -30°C and relative humidity above 70%), were used to test the effectiveness of the system against tiger mosquito eggs after 5, 10 and 15 days of egg-matrix contact. The term of comparison was the same suspensions of BB prepared for the growth experiment (G and H) while distilled water was used as a control which, having zero mortality, was not shown in the graph in figure 8. As further control, a solution of water and peptone 1 % by weight was used to evaluate any effects on hatching (K).

The concentration of Beauveria bassiana inside the matrices that will act against the eggs will be that reached after 24 days of incubation, i.e. for matrices of the order of 10⁷ conidia/mg, while 10⁵ conidia/ml for suspensions. Therefore, quantities of matrix and suspension will be used such as to have the same number of starting conidia.

The purpose of the test is to verify the "kill" functionality after verifying the "survival and growth" function. All tests were carried out under sterile conditions on 5-day-old tiger mosquito eggs collected at the insectarium of the La Sapienza University of Rome. T o carry out the test, 1 g of each matrix was spread on sterile filter paper (4x4 cm² and 1 mm thick) while 1.5ml of each suspension (G and H and K in figure 8, respectively G and H suspensions of BB prepared for the growth experiment and the control (K) carried out with a solution of water and peptone 1% by weight to evaluate any effects on hatching) and control based on distilled water were collected and uniformly dispersed on the surface of the sterile filter paper.

Three sterile papers were prepared for each matrix and suspension, one for each contact time (5, 10 and 15 days). On each of which 30 tiger mosquito eggs are placed. From this moment (To) 5, 10 and 15 days of egg-matrix contact are counted.

During contact, the matrices are stored in a Binder model KBF 115 climatic chamber (separated from any contaminants and thoroughly cleaned) at a temperature of 28°C and relative humidity of 80%. To evaluate the effectiveness, the samples thus prepared were subjected to a hatching test.

The same type of analysis was carried out on matrices prepared as in example 1 with concentrations of 2-FIEC by weight ranging from 2-30% but without Beauveria bassiana as an additional evaluation of the effect of the matrix without Beauveria bassiana.

Hatching test

After the expected contact time (5, 10, 15 days), the papers with the matrix and the eggs are moved with sterile disposable forceps into sterile plastic containers filled with 80 ml of distilled water and 1 mg of yeast (necessary to promote hatching by reducing oxygen in the water and as a source of nutrition for the larvae). The papers with the matrix and the eggs are kept in water for 5 days (average hatching time for the eggs) in an air-conditioned environment at 25°C (the temperature and humidity are monitored with the aid of a data logger). At the end of the 5 days the presence of viable larvae in the hatching water is assessed by means of a larval count. Furthermore, with the aid of a Leica series 230 and model 238 optical microscope, any larvae trapped inside the matrix are searched for and the presence ofstructuresis evaluated with a SEM EVO 40 electron microscope from Zeiss Beauveria bassiana on the eggs. Unhatched eggs (which are monitored for a further 10 days) and non-viable larvae trapped in the matrix are considered non-viable.

The viability of the eggs was preliminarily evaluated on the batch of eggs used for the test by carrying out a random hatching test on a group of 100 eggs performed with the same protocol reported.

The same type of test was performed on matrices prepared as in example 1 but without Beauveria bassiana with a concentration range of 2-30% by weight of 2-HEC and each containing 1% by weight of peptone in order to evaluate the effect itself of 2-HEC at various concentrations.

The histogram in figure 8 shows the corrected mortality values, evaluated as the number of viable larvae on the number of eggs placed on the matrix after 5, 10 and 15 days of egg-matrix contact. The reference control against which the corrected mortality was assessed is distilled water (0% mortality, not shown in the figure).

The results obtained shown in figure 8 are the average of 3 experiments, showing how mortality increases with the contact time for the matrices in which the peptone is present, while it decreases in the others. In the preferred form of preparation the corrected mortality is higher than 70% already at 10 days of contact. Therefore, the matrix satisfies the functions of "biocide survival and growth" and "kill". In the tested samples without BB, the results shown in figure 9 show that all cases satisfy the “kill” functionality. The yield stress curves and the threshold yield stress (i.e. the value above which the matrix can be used because it does not trickle) are also reported. For the matrices with concentration between 2-8% of 2-HEC the corrected mortality is between 80-90% but, analyzing the matrices before the hatching test it was verified that the eggs were already hatched inside the matrix and are death before the hatching test, therefore this event cannot be associated with a direct action of the matrix. Furthermore, it can be seen that between 2-8% of 2-HEC the yield stress value does not exceed the threshold value and therefore the matrix cannot be used on a vertical wall as required by the "lure" functionality.

For matrices at 16 and at 30% of 2-HEC there is a corrected mortality which reaches 90% for the 15 days of egg-matrix contact. At these concentrations the eggs do not hatch before the test and therefore mortality is associated with the mechanical action of the matrix.

As an example, figure 10 shows the images taken (A) with a Leica series 230 model 238 stereo microscope showing the entrapment action of the larvae by the matrix on newborn larvae (mechanical action) and (B) obtained with a scanning electron microscope (SEM EVO 40 by Zeiss) which shows the action of BB on the eggs that are colonized by the mycelium of the fungus.

In figure 10A the arrow indicates an example of a larva trapped immediately after hatching and dead as it is unable to move to find food and/or breathe. In image 10B we can see the presence of the BB structures on the mushroom.

Example 2: preparation of a matrix for the control of the tiger mosquito (Aedes albopictus) based on 2-HEC and Beauveria bassiana subjected to a cold freeze drying process.

An alternative form of preparation provides a matrix intended for the control of the tiger mosquito ( Aedes albopictus) with the following composition: distilled water, 20-300g/liter of 2-hydroxyethyl-cellulose (2-HEC, Sigma Aldrich) powder average molecular weight 90000, 10g/liter of peptone for mycology (Sigma-Aldrich), 10mg/liter of spores and mycelium of Beauveria bassiana strain CD 1123.

The protocol for the preparation of 120g of matrix at 16% by weight provides for 2-HEC (starting from sterile conditions of materials and instruments): a) prepare 100ml of sterile distilled water, b) add 1g of pepton and mix at room temperature until completely dissolved, c) add 1g of conidia and mycelium of Beauveria bassiana strain CD 1123 (corresponding to a turbidity of 2 Me Farland and an initial concentration of about 10³conidia/ml of suspension), d) mix at room temperature for 10 minutes, e) add 16g of 2-HEC and mix at room temperature for 30 minutes (estimated time for the complete gelling of the 2-HEC which occurs through a hydration reaction. Time is verified by analyzing the viscosity variation over time), f) pour or inject the prepared matrix into a mold of the desired shape (for example in the cavity of a hollow cylinder with an internal radius of 10 cm and external radius of 11 cm), g) freeze the matrix and the mold for 2 hours at -40°C, h) subjecting the frozen matrix to a lyophilization process at 0.3 milibars and -40°C of temperature for 12 hours until the complete elimination of the water by sublimation.

Biocompatibility test and conidic count on matrix subjected to freezedrying

A certain quantity of matrix prepared as in example 2 was taken and rehydrated until obtaining a matrix with 16% by weight of 2-HEC, that is, until a matrix with the same characteristics as that of example 1. This was incubated under the same conditions as the matrices made and tested in example 1, in order to assess whether the freeze-drying process had an impact on any of the matrix functions and in particular on that of " survival and growth ". It emerged that the process does not affect the growth of Beauveria bassiana in the matrix, in fact after 24 days the concentration of the biocide is at the same level as an untreated matrix, prepared as in example 1 (figure 11).

Another part of the matrix prepared as in Example 2 was taken from the same preparation and stored in a closed container at room temperature. Every 4 days a quantity of matrix useful to perform the conidic count test was taken as previously described in example 1. From the test it emerged that the lyophilized matrix maintains a constant conidic concentration for the entire duration of the test, so it can be preserved without losing vitality and therefore effectiveness (figure 11).

Therefore the freeze-drying process does not alter the functioning of the matrix. Example 3 (comparison): preparation of a matrix intended for the control of the tiger mosquito (Aedes albopictus) based on 2- hydroxyethylcellulose (2-HEC) cross-linked with poly-ethylene glycol- diglycidyl ether (PEGDE) and sodium hydroxide (NaOH).

One form of comparison preparation provides a matrix intended for the control of the tiger mosquito ( Aedes albopictus) with the following composition: distilled water, 20-300 g/liter of 2-hydroxy-ethylcellulose powder (2-HEC, Sigma Aldrich) average molecular weight 90000, 10g/liter of peptone for mycology (Sigma-Aldrich), 20g/liter of NaOH, 20g/l of PEDGE. The following procedure is aimed at preparing approximately 108g of cross-linked material, i.e. an 8% by weight gel of 2-HEC (if you want to reduce or increase the final quantity, reduce or increase all quantities proportionally).

1. Preparation of 100 g of a 0.5M NaOH solution (Solution A) a. Weigh 2g of solid NaOH (0.05 moles). b. In a suitably sized beaker weigh 10Og of distilled water. c. Add the NaOH to the solution until completely dissolved (clear solution).

2. Preparation of 100g of a 0.5M HCI solution (Solution B, to be used at the end of the process to neutralize and wash the cross-linked gel). a. Weigh 4.94g (or take 4.10ml) of 37% hydrochloric acid. b. Add the HCI to 96g of distilled water and mix until the acid is completely dissolved. 3. Preparation of the crosslinking solution (Solution C) a. Weigh 2g of PEGDE into a suitably sized beaker b. Add the 100g of solution A and mix until completely dissolved (clear solution).

4. Preparation of the gel a. Place the beaker containing solution C under a mechanical stirrer equipped with a suitable shovel. b. Stir the mechanical stirrer at a suitable speed c. Weigh 8g of 2-HEC and 1g of peptone and add them to the solution under constant stirring d. Incubate at a controlled temperature of 30°C for 5 hours 5. Wash the gel a. Remove the cross-linked gel from the beaker, cut it into pieces. b. Weigh 400g of distilled water into a suitably sized beaker c. Add 10Og of solution B to obtain final 500g d. Dip the gel into the washing solution and keep it stirring constantly for 2-3 days.

Oviposition test

A colony of tiger mosquitoes was reared in an insectarium placed at 26°C and 70% relative humidity and with a light hours: hours of darkness ratio 14:10. Mosquitoes had a blood meal 48h before laying (time necessary for the eggs to mature. At the end of the maturation time, 30 pregnant females were kept in cage. 2 plastic containers with 30 ml of distilled water are positioned inside the cage. The control (absorbent paper, C) was placed on the inner walls of the first container. The matrix (T) was positioned on the walls of the second container. After 24h the containers were recovered and the eggs were cashed. The test is was repeated 3 times with matrices having different concentrations of 2-HEC prepared as in example 3.

The histogram shows the values of T and C (already described above). The test shows how the presence of substances useful for crosslinking acts as a deterrent for the deposition of the eggs. Specifically, residues of hydrochloric acid and/or NaOH and/or PEGDE are repellent for the deposition of the tiger mosquito.

In fact, from the results shown in fig. 12 it emerges that the cross-linked gel described in example 3 is not attractive, on the contrary, it is totally rejected by insects. Failing the "lure" functionality the matrix is not functional.

Example 4: preparation of a matrix based on 2-hydroxyethylcellulose (2-HEC) and sorbitol, intended for the control of the tiger mosquito (Aedes albopictus). According to this embodiment, the matrix is intended for the control of the tiger mosquito ( Aedes albopictus) and has the following composition: distilled water, 20-300 g/liter of 2-hydroxyethylcellulose powder (2-HEC, Sigma Aldrich) average molecular weight 90000, 60g/liter of sorbitol (D-Sorbitol, Sigma-Aldrich) molecular weight 182.17.

In the preferred embodiment the procedure for the preparation to obtain about 120 g of matrix at 16% by weight of 2-HEC foresees (starting from the conditions of sterility of the materials and instruments): a) preparing 100 ml of sterile distilled water, b) add 6 g of D-Sorbitol and mix at room temperature until completely dissolved, c) add 16 g of 2-HEC and mix at room temperature for 40 minutes (estimated time for the complete gelling of the 2-HEC which occurs through hydration reaction. Time was verified by analysis of viscosity variation over time under standard conditions).

This preparation has pH values between 6-7 and salinity values below 3% under standard conditions (measured with a Mettler Toledo model pH8 benchtop pH meter and a Bormac COND 70+ portable conductivity meter, respectively). In this preparation, the presence of the humectant D-Sorbitol in a concentration by weight of 6% reduces the evaporation rate, maintaining for a longer time the conditions suitable for carrying out the various functions (linked to the quantity of free or bound water).

A tiger mosquito deposition test has been set up in the laboratory. In the test, the prepared matrix thus prepared was compared with another, prepared as reported here, but without D-sorbitol. Both were exposed to conditions suitable for spawning (25°C and 70-80% relative humidity). In the test the number of eggs present on the matrix was monitored weekly.

The test showed that both matrices are luring. On the matrix without sorbitol the number of eggs remained approximately constant after 3 weeks while on the matrix containing 6% of sorbitol the deposition continued until week 5, demonstrating the prolonged maintenance of the conditions suitable for the lure.

Field test: attractive and lethal effect of the gel on adults caused by mechanical entrapment and stickiness of the attractive system.

The test was carried out in the field in a small area (garden) in summer environmental conditions (average temperature 28°C average relative humidity 70%). The test was carried out on a matrix containing 16% by weight of HEC with the addition of 6% by weight of sorbitol (pH 6 and salinity <3%, carried out as reported in the preparation referred to in Example 4 and spread with a 3 mm layer on the side walls of a cardboard support. The results were recorded after 30 days in the field.

The gel exerts its attractive action without the presence of any chemical, semiochemical, food stimulant but only simulating (together with the cardboard structure) the conditions of humidity, lack of direct exposure and non-repellent composition. The insecticidal action is carried out by mechanical action on the adults (in this case pregnant females who have laid only a limited amount of eggs) and who have remained trapped in the gel consequently to the viscosity and adhesive effect of the humectant (sorbitol), the gel is free from any natural or chemical insecticide. Example 5: preparation of matrices based of 2-HEC with different salinity and pH values, intended for the control of the tiger mosquito (Aedes albopictus) and evaluation of the biomimetic attraction effect.

According to this embodiment, the matrix is intended for the control of the tiger mosquito ( Aedes albopictus) and has the following composition: distilled water, 20-300 g/liter of 2-hydroxyethylcellulose powder (2-HEC, Sigma Aldrich) average molecular weight 90000, variable content of NaCI, NaOH and HCI. In the preferred embodiment the procedure for the preparation to obtain about 120 g of matrix at 16% by weight of 2-HEC foresees (starting from the conditions of sterility of the materials and instruments): a) preparing 100 ml of sterile distilled water, b) Case 1 : add HCI or NaOH in quantities that make the pH vary with values of 10; 8; 5.5; 3, Case 2: add NaCI in such quantity as to vary the salinity from 0 (distilled water), 2, 3, 4% c) add 16 g of 2-HEC and mix at room temperature for 40 minutes (estimated time for the complete gelling of the 2-HEC which occurs through hydration reaction. The time was verified by analysis of the viscosity variation over time under standard conditions).

The pH and salinity values are measured with a Mettler Toledo brand bench pH meter model pH8 and a Bormac model COND 70+ portable conductivity meter respectively. An oviposition test was carried out as in example 1 on the matrices prepared as described. From the results reported in Figure 14 and Figure 15, it emerges that outside the salinity and pH ranges reported previously, the biomimetic attraction effect on the tiger mosquito disappears, making the matrix ineffective.

In the prior art, salinity or pH ranges are not clearly reported and the effects of their variation on the attractiveness towards insects are not documented, which on the contrary are attracted through the use of semiochemical substances (pheromones or attractants). Therefore, the known art cannot solve the technical problem of attracting insects without the use of attractants or pheromones or baits.

Example 6: preparation of a matrix based on 2-hydroxyethylcellulose (2-HEC) and lactic acid as a humectant, intended for the control of the tiger mosquito (Aedes albopictus).

A matrix with 16% by weight of HEC was made with 16 g of HEC (polymer) in 100 g of distilled water as in example 5, substituting lactic acid as humectant (respectively with a concentration of 0.01 ; 0.1 ; 1 ; 3; 5% by weight) with sorbitol was made and tested during the laying phase with the same method reported in example 1. The pH of the matrix was also evaluated with the variation of the lactic acid concentration. The pH and salinity values are measured respectively with a Mettler Toledo model pH8 benchtop pH meter. Also in this case it emerged (Fig. 16) that for acid pH there is no deposition. In this case the attractive functionality fails due to the choice of the humectant.

Example 7: preparation of matrices based on sodium alginate ( SA ) and carboxymethylcellulose (CMC) with and without the addition of sorbitol as humectant, intended for the control of the tiger mosquito (Aedes albopictus). According to this embodiment, the matrices are intended for the control of the tiger mosquito ( Aedes albopictus) and represent alternatives to the preferred composition based on HEC. The gels have the following composition: distilled water, 20-300 g/liter of sodium alginate powder (SA, Sigma Aldrich) average molecular weight 120000, 20-100 g/liter of carboxymethylcellulose powder (SA, Sigma Aldrich) weight molecular average 250000. To this composition sorbitol can be added as a humectant, 60g/liter of sorbitol (D-Sorbitol, Sigma-Aldrich) molecular weight 182.17.

In the preferred embodiments, the procedure for the preparation to obtain about 120 g of matrix at 16% by weight of SA provides (starting from conditions of sterility of the materials and instruments): a) prepare 100 ml of sterile distilled water, b) add 6 g of D-Sorbitol and mix at room temperature until completely dissolved for the preparations to which it is added, c) add 16 g of SA and mix at room temperature for 40 minutes (estimated time for the complete gelling of the SA which occurs via hydration reaction. The time has been verified by analyzing the viscosity variation over time under standard conditions).

The preferred preparation based on CMC foresees the same preparation steps and a quantity of CMC to be added equal to 5g per 100 ml of water to obtain a CMC gel at 5% concentration by weight. These preparations have pH values between 6-7 and salinity values below 3% under standard conditions (measured respectively with a Mettler Toledo model pH8 benchtop pH meter and a Bormac model COND 70+ portable conductivity meter).

The same characterization tests reported in example 1 and shown in Figure 13 (panels a-d) were performed on the matrices thus created (yield test, viscosity values as the polymer concentration varies, weight loss over time, quantity of free water ), in order to verify that these values of the gels produced fall within the required ranges. The results confirm that the values are within the expected ranges to perform biomimetic lure, be lethal and allow the growth and proliferation of a biopesticide, as well as to be functional for an adequate period of time to obtain a technical advantage over the solutions currently in use for the control of the tiger mosquito (eg masonite rods).

Verification test of biomimetic lure functionality in the cage (Fig. 17) and in the field (Fig. 18) on the matrices based on SA and CMC with and without sorbitol.

A tiger mosquito deposition test was set up in the laboratory using the matrices prepared as in example 7 as a deposition substrate. Both were exposed to conditions suitable for spawning (25°C and 70-80% relative humidity). In the test, the number of eggs present on the matrix at that time was monitored weekly. The test revealed that the matrices are capturing (Figure 17) at levels comparable to those of the HEC-based matrix. On the matrix without sorbitol the number of eggs remained approximately constant after the first week while on the matrix containing 6% of sorbitol the deposition continued until week 3 demonstrating the prolonged maintenance of the conditions suitable for the lure.

A field test was performed, randomly distributing the matrices containing 6% sorbitol based on HEC16%, SA16% and CMC5% (prepared as reported) in the perimeter of a garden. It emerges (Figure 18) that even in field conditions the attractiveness is confirmed.

Biocompatibility test and insecticidal activity of gels without bioinsecticide but still able to host it (Fig. 19).

The biocompatibility towards the biopesticide Beauveria bassiana of matrices based on FIEC16%, SA16% and CMC5% containing 6% by weight of sorbitol and their insecticidal effect was tested in a test performed as reported in example 1 (test of biocompatibility and lethality on Aedes albopictus eggs by hatching test). The hatching test on eggs Aedes albopictus previously reported in Examplel , already reported the insecticidal activity of the gel without bioinsecticide but still able to host it. It emerged (Fig. 19) that all the compositions have the ability to host and proliferate the Beauveria bassiana for at least 24 days. They also have an insecticidal capacity (Fig. 19) which is completely independent of the presence of the bioinsecticide. Therefore, the gel has a lure and kill action completely independent of the presence of food and semiochemical attractants (eg pheromones of any form), but linked exclusively to the properties of the polymer which specifically fall within the ranges shown.

Example 8: Caged matrix preference test: Deposition of Aedes albopictus in a cage over a three week period (Fig. 20). The matrices based on FIEC, SA and CMC with concentrations of 16% by weight (FIEC and SA) and 5% by weight (CMC) respectively with and without the addition of 6% by weight of sorbitol as humectant were produced as reported in examples 1 , 4 and 7. They were subjected to an oviposition test on aedes albopictus carried out as reported in example 1 to evaluate both the attractiveness of the different substrates in absolute terms and over time of the gels (T = number of eggs laid on gel) compared to masonite as control (C = number of eggs laid on control). From the test (Fig. 20) it emerged that all matrices are attractive, in particular they are more attractive than masonite and in the first week have comparable attractiveness. In the following weeks, the masonite ceases to be attractive (T/C values remain constant) since it dehydrates while the CMC added with sorbitol is more attractive (higher T/C value). The result can be explained by the lower dehydration of CMC 5% + Sorbitol 6% as shown in Fig. 13 panel C.

Example 9: Aedes albopictus egg deposition on viscous sodium alginate solution at 16% by weight after gelation with calcium chloride.

The deposition test was carried out as the one previously described in example 1 in the section of the oviposition test for evaluation of the biomimetic "lure" functionality, and concerned non-crosslinked (SA) and calcium chloride crosslinked (SA-CaCl2) sodium alginate gels prepared as follows. The non-crosslinked solution of sodium alginate (SA) tested in the deposition phase, in the most general form was made as follows: distilled water, 20-300 g/liter of medium viscosity sodium alginate powder (sodium alginate medium viscosity, Sigma Aldrich). The preparation procedure is the same as in the previous examples (example 7). The crosslinking with calcium chloride of the SA16% matrix prepared as reported is as follows and is carried out directly on a masonite support using a solution of calcium chloride (SA- CaClsamples₂) with the following procedure.

1. spread a 3mm layer on a 2x10cm masonite strip,

2. prepare of a 0.05 molar calcium chloride solution in distilled water,

3. immerse the substrate coated with alginate inside the calcium chloride solution.

4. bring the solution to a boil and keep it for 2 minutes. The masonite support coated with calcium chloride cross-linked alginate was used in the deposition test. The preparations have pH values between 6-7 and salinity values below 3% under standard conditions (measured respectively with a Mettler Toledo model pH8 benchtop pH meter and a Bormac model COND 70+ portable conductivity meter) and viscosity values between 5-10 Pas (for SA) while SA-CaCUOkPas and module G ^* equal to 0.1 MPa.

Deposition results

Through the example reported it is shown (in Fig. 21) how starting from an attractive gel based on biopolymer (which is also an example of an alternative attractive composition), varying its gelation properties with the same salt reported in literature (WO2011/123760), the attractiveness is lost. The purpose of the insecticide matrix in the invention, that is to attract and kill the insect even without bioinsepticide, cannot be achieved if the lure functionality is inactive. As shown, even partially respecting the range of parameters that define it, the gel is not attractive due to its composition. Therefore, its realization cannot be obtained in a trivial way by combining what is already present in the known art. Example 10: Verification of the attractive functionality on other insects and analysis of the attractive conditions.

The matrices produced as reported in examples 1 and 4 and 7 were used to verify lure and kill functionality on other insects, using different concentrations based on the need to evaluate other attractive conditions for the tested insects.

Culex mosquito

A cage deposition test was carried out as described in example 1 but using culex mosquitoes. The comparison was carried out on both HEC and SA with concentrations from 2 to 12% as reported in Tab 1. The viscosity and free water values contained are reported. Differently from the tiger mosquito (Aedes albopictus) the deposition values are higher (compared to the water which is the control) on less viscous gels and with higher % of free water. This is in line with the behavior of the culex mosquito which lays its eggs directly in water and not on humid substrates (e.g. masonite) as the tiger mosquito does and demonstrates the validity of the biomimetic approach.

T able 1 T = Eggs laid on tested gel, C = Eggs laid on control

Mites (Dermanyssus gallinae)

A test in a cage was carried out (with 30 specimens of Dermanisss Gallinae), to evaluate the preference of the substrate in the resting phase following the blood meal. This type of mite naturally retreats in humid places after the blood meal (usually damp cardboard or sponge present in animal eg poultry cages). The mortality of the insecticide-free substrate (understood as dead mites of those who chose it as a resting place) was also evaluated. In this case, wet cardboard was used as a control and gels were tested with increasing concentrations of HEC and CMC containing 5% by weight of sorbitol as a humectant. The increasing concentrations are not the same, while the viscosities are comparable (viscosity and free water content are reported in the table). It has emerged that the best results in terms of attractiveness and lethality are obtained for values between 11 and 16 Pas with percentages of free water around 80%. Considering mortality, the best compromise is that with 10% HEC (viscosity = 11 Pas and % free water = 83% with a T/C = 3 and corrected mortality equal to 90%) Table 2 T = Eggs laid on tested gel, C = Eggs laid on control

^* Number of insects that have preferred the gel/number of insects over the control (wet cardboard), for values greater than 1 the gel is preferred.

Example 11 : comparison between patent US 2011/0184040 and proposed insecticide device.

A test was carried out to assess whether with an aqueous gel, free of insecticide and/or attractive substances, made with the formulation reported in US patent 2011/0184040, the conditions required to achieve the technical solution to the problem of attraction of insects without bait and/or food or semiochemical attractants and the advantages reported by the authors and associated with the insecticidal matrix described.

In particular, HEC-based gels were made with the same viscosity characteristics reported for the aqueous gel (0.01-10Pas measured with Malvern Kinexus rotational rheometer with shear rate = 100s¹ shown in figure 22) containing sorbitol as humectant and corresponding to the preferred formulation of the so-called aqueous gel reported in US patent 2011/0184040 (paragraph 10). The formulations were applied by spraying 2ml of aqueous gel on a 1 cm² panel (equal to the highest quantity foreseen in the comparative patent which is 201/ha dispersed on the smallest spot surface expected). Under these conditions the gels showed a drying below the masonite levels (and therefore not attractive for the tiger mosquito and as reported in example 10 not even for culex or mites) already after 3 days (as reported in Table 3). In addition, the total absence of a viscous layer of gel prevents the lethal effect by entrapment on both larvae and eggs as well as the possibility of growing a bioinsecticide. It is evident that the aqueous gel proposed in the patent does not have any type of attractive effect if not used in a formulation containing at least one attractive substance and that it cannot guarantee any of the advantages reported (environmental safety, durability, biocompatibility, etc.) in the document, let alone solve the same technical problem that is to be solved with the present invention.

Although in patent US2011/0184040 reference is also made to a concentrated gel (with viscosity up to 100Pas) and other viscoelastic properties partially superimposable to the gels described here, this is explicitly described as inapplicable for spraying (par.17) and therefore does not concern the solution of the problem of the control of the artopods, which in the text is carried out only with the aqueous gel. Furthermore, both the aqueous gel and the concentrate from which it is derived by dilution with water, explicitly and without exception contain insecticides (exclusively synthetic and not biopesticides) and attractants and therefore cannot solve the technical problem proposed in the present invention, on the contrary if deprived of these elements from the composition they would not work. Finally, both the aqueous and the concentrated gel do not provide for any control of salinity or pH which are neither explicitly indicated nor evaluated.

Therefore, the transition from the gel proposed in patent US201 1/0184040 to the one described here cannot be trivially obtained by resuming the gel described (aqueous or concentrated), partially varying the formulation and using it in a different context, as for the concentrated one they are not known a priori some of its properties considered essential for the biomimetic lure and for the biocompatibility of a biopesticide (eg pH and salinity) while for the diluted one there are no adequate technical conditions.

Table 3: Weight loss of gels with viscosities comparable to the preferred formulations in prior art and comparison with masonite (definitely attractive substrate).

Claims

1. Use of a biomimetic insecticide matrix in the form of gel for hematophagous insects having at the same time the functionality of "biomimetic lure and kill", said matrix comprising one or more biocompatible and biodegradable polymers chosen from: agar, pectins, polysaccharides, chitosans, alginates, cellulose for example the hydroxymethyl- and hydroxyethyl-cellulose, the carboxymethyl- and carboxyethyl-cellulose said matrix having a pH between 4 and 8, salinity not higher than 3%, a quantity of free water higher than 0, preferably between 5- 98% by weight and a viscosity between 10³-10⁴ Pas.

2. Use according to claim 1 , wherein the matrix also has the function of survival and growth of biopesticides inside it, said biopesticides being active against bloodsucking insects.

3. Use according to the preceding claim in which the biopesticide is Beauveria bassiana.

4. Use of the matrix according to any one of claims 1-11 as an insecticidal agent against bloodsucking insects which has a biomimetic lure and kill action with and without biopesticides.

5. Biomimetic insecticide matrix in the form of a gel for hematophagous insects having at the same time the functionality of "biomimetic lure and kill" and that of survival and growth of biopesticides inside it active against bloodsucking insects; said matrix comprising one or more biocompatible and biodegradable polymers selected from: agar, pectins, polysaccharides, chitosans, alginates, celluloses such as hydroxymethyl- and hydroxyethyl-cellulose, carboxymethyl- and carboxyethyl-cellulose said matrix having a pH between 4 and 8, salinity not higher than 3%, a quantity of free water higher than 0, preferably comprised between 5-98% by weight and a viscosity comprised between 10³-10⁴ Pas.

6. Insecticidal matrix according to claim 5, further characterized by having yield stress values between 0-2000Pa, storage modulus and/or loss modulus values between 0-2MPa.

7. Insecticidal matrix according to any one of claims 5-6, in which the amount of free water is comprised between 5-98% by weight.

8. Insecticidal matrix according to any one of claims 5-7, in which the yield stress value is not less than 50Pa, the free water content is in the range 70-80% by weight and the viscosity is not less than 2 Pas.

9. Insecticidal matrix according to any one of claims 5-8, which maintains at least 30% of the initial weight within the fifteenth day of exposure to conditions of temperatures of 18-30°C and humidity of not less than 70%.

10. Insecticidal matrix according to any one of claims 5-9, which is active against insects selected from: horseflies, horse flies, mosquitoes, including C. tarsalis, C. pipiens, A. aegypti, A. sierrensis, A. nigromaculis, A. albimanus, A. albopictus, bedbugs, papadaci, fleas, ticks, lice, mites including Dermanyssus gallinae, nematodes, annelids, their larvae and eggs.

11. Insecticidal matrix according to any one of claims 5-10, which is active against bloodsucking insects both comprising a biopesticide and whether it does not.

12. Insecticidal matrix according to any one of claims 5-11 , which further comprises a biopesticide selected from: Gram-negative and

Gram-positive bacteria, actinomycetes, fungi, protozoan yeasts, algae and their spores, in particular Bacillus subtilis subsp. Krictiensis, Pseudomonas pyrocinia, Pseudomonas fluorescence, Gliocladium wirens, Trichoderma reesei, Trichoderma harzianum, Trichoderma hamatum, Trichoderma viride, Streptomyces cacaoi subspecies asoensis; or microorganisms such as Bacillus thuringiensis, or entomopathogenic fungi that belong to four main groups: Oomycota, including Lagenidium and Leptolegnia, Ascomycota, including Aspergillus, Beauveria bassiana, Metarhizium, Paecilomycora, and Zygomycota, including Conidiobolus, Entomniaga, Entomniaga Furia, Massospora, Neozygites, Pandora, Zoophthora.

13. Insecticidal matrix according to the preceding claim in which the biopesticide is Beauveria bassiana.

14. Insecticidal matrix according to any one of claims 5-13, which further comprises a wetting agent selected from: glycerin, propylene glycol, glyceryl triacetate sorbitol, xylitol, maltitol, polydextrose, natural extracts such as quillaia, compounds such as lactic acid or I 'urea.

15. Insecticidal matrix according to any one of claims 5-1 , which does not contain food, semiochemical actives and related mixtures.

16. Insecticidal matrix according to any one of claims 5-15, which is in the form of gel, spreadable paste or other hydrated or dehydratable and rehydratable moldable form.

17. Method for preparing the matrix according to any one of claims

5-16 comprising the basic steps of: in a temperature range of 15-60°C preparing an aqueous solution and adding the polymer until gelation, continuing to mix at constant temperature.

18. Method according to the preceding claim, in which a solution containing a biopesticide and optionally a wetting agent is added to the mixture of polymer and water.

19. Method according to any one of claims 17-18 wherein the gel obtained is subsequently subjected to elimination of water, for example by lyophilization.

20. Insecticidal device comprising the matrix according to any one of claims 5-16.

21. Use of the device according to claim 20, as an insecticide to be placed inside closed places such as: homes, offices, hospitals, and in open environments such as: parks, neighborhoods or cities.

22. Use according to the preceding claim in the civil, veterinary, zootechnical, industrial and agricultural fields.

23. Products in which the matrix according to any one of claims 5-16 is spread on a support and protected with a sealed envelope.

24. Kit comprising the articles according to the preceding claim on which the matrix is spread in a sealed envelope and/or a portion of said matrix is in dehydrated form to be rehydrated, and instructions for use.

25. Kit according to the preceding claim wherein the matrix is selected from matrices comprising or not comprising a biopesticide.