NUTRITIONAL SOLID COMPOSITIONS WITH ACARICIDE ACTIVITY FOR APICULTURE AND THEIR USE FOR THE PROPHYLAXIS AND TREATMENT OF VARROA INFESTATIONS

Abstract

The invention concerns a solid feed composition for use as nourishment for bees and for the prevention and treatment of acariosis, and, in particular, of infestation by Varroa destructor, as well as the relative treatment method, comprising: a) nutritional and tonic ingredients, consisting of algae containing vegetal proteins and yeasts; sugars and lower organic acids; b) natural antioxidants and antiseptics contained in the extracts of Origanum vulgare and of Pelargonium graveolens or essential oil of geranium and in the essential extracts of one or more aromatic or medicinal plants selected from: Crocus sativus, Monarda citriodora, Melissa officinalis, Myristica fragrans, and Origanum majorana; and c) curative substances for bees, comprising at least one of thymol and essential extracts of Thymus vulgaris, and at least one of oxalic acid, extracts of Aloe vera or Aloe arborescens, geraniol and extracts of Beta vulgaris cv. altissima, and mixtures of two or more of the same.

Claims

1. A solid nutritional composition for use as bee nutrition and, at the same time, for the prophylaxis and treatment of acarioses, in particular of Varroa destructor infestations, comprising the following ingredients: a) nutritional and tonic ingredients, comprising: algae containing vegetal proteins, alone or in combination with yeasts; sugars and lower organic acids; b) natural antioxidants and antiseptics contained in the essential extracts of Origanum vulgare and Pelargonium graveolens or geranium essential oil, and in the essential extracts of one or more other aromatic or medicinal plants selected from: Crocus sativus, Monarda citriodora, Myristica fragrans, and Origanum majorana; and c) curative substances for bees, comprising at least one of thymol and essential extracts of Thymus vulgaris, and at least one of the following:_oxalic acid, extracts of Aloe vera or Aloe arborescens, geraniol and extracts of Beta vulgaris cv. Altissima, and mixtures of two or more of the same.

2. A solid nutritional composition for the use according to claim 1, wherein said nutritional and tonic ingredients consist of: algae containing vegetal proteins and brewer's yeast; dextrose, glucose and/or fructose; acetic, citric and/or tartaric acid.

3. A solid nutritional composition for the use according to claim 2, wherein said algae are selected from spirulina, kelp, Klamath, chlorella and mixtures thereof.

4. A solid nutritional composition for the use according to claim 1, wherein said nutritional and tonic ingredients also comprise sources of fatty acids, preferably lecithin, or a mixture of oils, or a mixture of oils and lecithin.

5. A solid nutritional composition for the use according to claim 1, wherein said nutritional and tonic ingredients also comprise sources of sterols and/or essential oils.

6. A solid nutritional composition for the use according to claim 1, wherein said nutritional and tonic ingredients also comprise glycerin.

7. A solid nutritional composition for the use according to claim 1, wherein said essential extracts containing natural antioxidants and antiseptics comprise essential extracts of Origanum vulgare and Pelargonium graveolens or geranium essential oil, Crocus sativus extract, Myristica fragrans extract and Origanum majorana extract.

8. A solid nutritional composition for the use according to claim 7, wherein said essential extracts containing natural antioxidants and antiseptics further comprise essential extracts of Monarda citriodora.

9. A solid nutritional composition for the use according to claim 8, wherein said essential extracts containing natural antioxidants and antiseptics further comprise essential extracts of Melissa officinalis.

10. A solid nutritional composition for the use according to claim 9, wherein said essential extracts containing natural antioxidants and antiseptics further comprise extract of dry roots of Polygonum cuspidatum.

11. A solid nutritional composition for the use according to claim 9, wherein said substances medicinal to bees also comprise mycelium extracts of fungi of the phylum Basidiomycota.

12. A solid nutritional composition for the use according to claim 9, wherein said substances medicinal to bees are Thymus vulgaris extract, Aloe vera extract, geraniol and Beta vulgaris cv. Altissima extract.

13. The solid nutritional composition for the use according to claim 12, wherein said solid food is in form of patties and also contains, preferably, one or more of emulsifiers, lecithin and vegetal oils.

14. A method of treatment of beehive colonies for the prophylaxis and treatment of acariosis through the administration of nutritional and medicinal substances, which method consists of using a product in powder, or in patties, having the following composition: a) nutritional and tonic ingredients, comprising: algae containing vegetal proteins, alone or in combination with yeasts; sugars and lower organic acids; b) natural antioxidants and antiseptics contained in the essential extracts of Origanum vulgare and of Pelargonium graveolens or geranium essential oil, and in the essential extracts of one or more other aromatic or medicinal plants selected from: Crocus sativus, Monarda citriodora, Melissa officinalis, Myristica fragrans, and Origanum majorana; and c) substances medicinal to bees, comprising at least one of thymol and extracts of Thymus vulgaris, and at least one of the following: oxalic acid, extracts of Aloe vera or Aloe arborescens, geraniol and extracts of Beta vulgaris cv. Altissima, and mixtures of two or more of the same.

15. The method of treatment of beehive colonies according to claim 14, wherein said nutritional and tonic ingredients consist of: algae containing vegetal proteins, preferably selected from spirulina, kelp, Klamath, chlorella and mixtures thereof, and brewer's yeast; dextrose, glucose and/or fructose; acetic, citric and/or tartaric acid.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0040] Therefore, the present invention specifically provides a solid nutritional composition, i.e. in powder or in patties, for use as nourishment for bees and for the prophylaxis and treatment of acariosis, in particular of Varroa destructor infestations, comprising the following ingredients: [0041] a) tonic and nutritious ingredients, mainly consisting of: algae containing vegetal proteins, alone or in combination with yeasts; sugars and lower organic acids; [0042] b) natural antioxidants and antiseptics contained in the essential extracts of Origanum vulgare and Pelargonium graveolens or geranium essential oil, and in the essential extract of one or more other aromatic or medicinal plants selected from Crocus sativus, Monarda citriodora, Melissa officinalis, Myristica fragrans and Origanum majorana; and [0043] c) curative substances for bees, comprising at least one of thymol and essential extract of Thymus vulgaris, and at least one of oxalic acid, extracts of Aloe vera or Aloe arborescens, geraniol and extracts of Beta vulgaris cv. Altissima and mixtures of two or more of the same.

[0044] Other features and preferred embodiments of the solid compositions for use according to the invention are set out in the dependent claims.

[0045] By comparing the solid product which is proposed for use as a feed for beekeeping and/or as an acaricide agent according to the invention with the liquid product of the prior art, it is to be noted that the strengths of the solid (powder) product are the following: [0046] Durability of the product (in liquid form the product has a shelf life of 18 months, while in powder form the shelf life is 30 months); [0047] Greater ease and economy of transport; [0048] Greater ease of administration, avoiding the need for water or other liquids, such as sugar molasses or syrups; [0049] Less time and lower labor costs to distribute the product in the hive, as it is a ready-made product and does not require dilution operations with control of the water percentages in solution, weighing, or problems of finding a source of water; [0050] Greater durability of the product inside the apiary; [0051] Possibility for bees to consume the product in a selective manner: in fact, in the liquid form the key ingredients are completely solubilized and cannot be separated, while the granular/powder form gives the bees the opportunity to consume the product according to the needs of the season and according to specific needs. In the tests carried out and in the numerous videos that were made to study how the bees consume the powdered product, it has been found that when the bees want to take more carbohydrates they use the ligula and labial glossa (with probable increase in the production of enzymes from the labial and thoracic glands located in the anterior ventral portion of the thorax) to dissolve the sugary substances, whereas when they require greater protein content they tend to bite and crush the granules of the powdered product with the mandibular apparatus and the support of the two mandibular glands; [0052] Greater flexibility and control of the Substances and ingredients to be assumed by the bees of the three castes (workers, drones, queen) through the average size of the granules of the powdered product (particle size), which can range from 10 to 1000 m (similar to the average size of pollen in nature). Furthermore: [0053] a) under 10 m size the powder product shows drawbacks such as compaction, and in general it is more prone to absorption of moisture and tither liquid products present in the beehive, with acceleration of the degradation of the product quality; [0054] b) for particle sizes exceeding 1000 m there may be problems in the homogenization of the basic ingredients of the formulation, with concentration gradients not uniformly distributed; also, a greater difficulty for bees in eating the product due to the weight of the single granule is possible, as well as a difficulty in handling the granule itself during consumption.

[0055] As already noted in the previous document, in general the substances that can be used in the preparation according to the invention are as follows:

A. Nutrient and Tonic Substances:

[0056] Yeasts, of which the bees are very fond, as for example brewer's yeast, which is very rich in vitamins B, are preferably added to amino acids of vegetable origin contained in algae. According to some preferred embodiments of the invention, said algae are spirulina (Arthrospira platensis), kelp (Laminariales), Klamath (Aphanizomenon flos-aquae) or chlorella (Chlorella vulgaris). The protein content of these algae is about 50%, with a fat content of about 7%.

[0057] A proper blending of protein sources is essential to ensure the appropriate percentages of amino acids that are essential for the development of larvae and adult individuals. In addition to being essential for their amino acid contribution, algae are used for their supply of micronutrients, minerals (up to 1.5% in dry weight of the algae), iron and other metals and vitamins (up to 2% by dry weight of the algae).

[0058] In particular, it has been noted that the addition of proteins of vegetal origin such as algae and brewer's yeast instead of animal proteins in the formulation results in a triple advantage: a) a component (i.e. milk) is eliminated which, due to the content of caseins, is subject to rancidity; b) the protein content contained in the feed significantly increased; c) the palatability of the product for bees is improved, as the bees consume the product better than the corresponding product with animal proteins. In addition, a possible toxicity to bees due to the use of an animal source of proteins is eliminated, and a product having an amino acid pattern similar to that of royal jelly and of pollen is obtained in terms of percentages and types of amino acids present. Moreover, a formula is obtained which can be used in powder form for the preparation of patties or cake.

[0059] To increase the palatability of the protein feed to be administered, sugars of the glucose, dextrose or fructose type. Actually, it is possible to add pollen and/or icing sugar (sucrose) in an amount ranging from 5% to 15% on the total dry weight of the formulation. As nutrients vitamin E, and essential amino acids extracted from Aloe arborescens or Aloe vera have also been used, which are listed below among the substances having curative activity.

[0060] Acetic acid and/or lower carboxylic or dicarboxylic acids C.sub.2-C.sub.6. Acetic acid, tartaric acid and citric acid have the ability to cleave the molecules of sucrose, promoting assimilation by the bees. Also, said acids are antifungals useful in combating the presence of the fungus Nosema and other fungal forms. Specifically, the formulation may contain acetic acid at a maximum concentration of 6% by weight, that helps to have a product pH below 7. A source of ascorbic acid (vitamin C), such as, for example, lemon juice (which contains the same, in addition to the most abundant citric acid), may be included in the formulation as a vitamin compound, as well as an anti-oxidant.

[0061] Sources of fatty acids: lecithin, or a mixture of oils or a mixture of oils and lecithin, as a source of short, medium and long chain fatty acids may be added. For example, ricinoleic, tricosanoic, myristic, myristoleic, linoleic, palmitic, palmitoleic, lauric acids, in which the correct ratio of omega 6 to omega 3 fatty acids must be set to at from 0.01% to 3%

[0062] Sources of sterols: for example, a combination of 24 methylene cholesterol, campesterol, beta-sitosterol, cholesterol (0.01% to 4%), canola oil derivatives, borage oil (Borago officinalis), Echinum spp. oil, turnip seed oil derivatives may be used.

[0063] Essential oils, or tpeircomponents, which increase the palatability of the product can be added, such as hydroxycinnamic acid, quercetin, rutin, narigenin, p-coumaric acid, contained in the essential oil of lemon balm and in the essential oil of monarda.

[0064] In the case of use as a candy products/patties: stabilizers for patties, such as emulsifiers, lecithin and vegetable oils (coconut oil and others) should be added.

B. Highly Active Antioxidants and Antiseptics

[0065] Essential oils with a high content of antioxidants and/or antiseptics are added to the formulation, such as carotenoids of the type of crocetin, crocin and picrocrocin extracted from flowers and/or stigmas of saffron (extracts of Crocus sativus); essential oils as limonene, e.g. extracted from lemon; geraniol, citronellol, terpineol and linalool, extracted from Pelargonium graveolens (geranium), or from Monarda citriodora var. citriodora; myristicin, elemicin, geraniol and/or safrole and other aromatic ethers extracted from Myristica fragrans (nutmeg); carvacrol, thymol and other minor phenols extracted from Origanum vulgare (ssp hirtum); and terpenes such as terpineol, borneol, sabinene and linalool, extracted from Origanum majorana.

[0066] Said active ingredients can be extracted, for example, from the plant species mentioned above and usually have a purity exceeding 55%, or they can be made synthetically.

[0067] Another advantageous antioxidant of vegetable origin is obtained from the dried roots of a plant of the polygonaceae, Polygonum cuspidatum, from which a 98% resveratrol extract can be obtained. This powerful antioxidant is also present in other plants belonging to the genera Polygonaceae and Vitaceae.

C. Other Therapeutic Substances

[0068] To counteract and/or prevent mites, as well as harmful fungal forms, viruses including the IIV6 and IAPV and the undesired side effects of neonicotinoids, there were used, in alternative to thymol produced by synthesis, the biologically active substances contained in the essential oil of Thymus vulgaris (common thyme) of the types red thyme essential oil (or oil of first distillation) and white thyme essential oil (or oil of second distillation). Thyme, whose essential oil is widely used in beekeeping, contains the two phenolic compounds with biocidal activity thymol (very active against Varroa) and carvacrol. as well as cineol, borneol, menthone, pinene, geraniol, alpha-terpineol and other terpene compounds.

[0069] In addition, substances contained in the essential oil of Aloe vera, or in the essential oil of Aloe arborescens (a species of less widespread aloe but richer in active biological ingredients) can be used. Aloe vera contains, similarly to Aloe arborescens, many biologically active compounds, including acemannan (a mucopolysaccharide known for its immunomodulatory activity, with antiviral action), cinnamic acid (germicidal, fungicide, analgesic), crysophanic acid (antimycotic), anthraquinones, including aloin (bactericide) and emodin (antiviral); beta-sitosterol, in addition to salicylic acid (anti-inflammatory) isobarbaloin (analgesic), socaloin, capaloin and barbaloin (anti-bacterial). It should be noted that the extracts of this plant also contain all essential amino acids and vitamin E.

[0070] Moreover, according to the invention, a good efficacy has been found in the use of extracts of the common beet or Beta vulgaris cv. altissima (sugar beet), which contains betalains (red pigments, attractants for bees), flavonoids, trimethylglycine (betaine), compounds with antioxidant activity, oxalic acid and vitamins belonging to group B.

[0071] Other medicinal substances can be added by adding geranium essential oil, which is extracted from geranium (Pelargonium) flowers and leaves and has geraniol as its main component, which is a terpene alcohol active as an antiseptic or antibacterial, as well as borneol, citronellol, linalool, terpineol, limonene, pinene and -methyl-eugenol, all of which are active antioxidants. Alternatively, only the chemical compound geraniol can be used.

[0072] Further, oxalic acid can be added to said extracts in amounts not higher than 0.1% of the total formulation,(to increase the disinfectant effect in synergy with the mentioned natural compounds.

[0073] Extracts (for example, alcoholic extracts) of fungal mycelium may be added, e.g. from fungi belonging to the phylum of the Basidiomycetes (Basidiomycota), order Polyporales, which have been shown to have antiviral properties against several viruses associated with the presence of varroa in the hive.

[0074] The following species may also be used: Fomes fomentarius, Ganoderma applanatum, Ganoderma resinaceum and Trametes versicolor, with final proportions from 0.01% to 1% by weight.

[0075] As regards the points that the nutritional product in powder form for apiculture according to the present invention has in common with the aqueous solution or aqueous suspension of the prior art, it is to be noted that the powder product, in being metabolized by the bees, does not lose its unique characteristics of:

1) Countering the poisoning by herbicides, such as paraquat, or pesticides of the neonicotenoid family of various generations. With regard to paraquat, it has been found that the solid product according to the invention (BeesVita Plus) reduces the amount of ROS formed by the herbicide, thus showing for the first time that a nutritional system can reduce the adverse effects of pesticides. It should be noted that, among the ingredients responsible for this unique effect, stand out, both singly and for their synergistic effects with one another, the antioxidant and antiseptic compounds with high activity contained in the essential extracts of: [0076] Origanum vulgare: contains oreganol, carvacrol and thymol, active against varroa. [0077] Origanum majorana: contains carvacrol, thymol, oreganol, pinene, sabinene, terpineol, terpinene, camphene, as well as tannins, flavonoids (apigenol, borneol), rosmarinic acid and caffeic acid, which is very important for countering the side effects of neonicotinoids. [0078] Myristica fragrans: contains triterpenes (camphene, pinene), monoterpenic alcohols (borneol, terpineol), cymol, dipentene, eugenol, geraniol, linalerol, sapol, safrole and myristicin. The monoterpenoid extracts which include terpinen-4-ol, -terpineol and 4-allyl-2,6-dimethoxyphenol have shown particularly strong antioxidant activities, and activities to counteract the effects of pesticides based on neonicotinoids. Three antifungal lignans were isolated and identified as eritro-austrobailignan-6 (EA6), meso-dihydroguaiaretic acid (MDA) and nectandrin-B (NB).
2) Raising the immune defenses of bees in general, with increased gene expression. For the first time, a nutritional system has been able to limit the deleterious effects of a pesticidea great advance in improving the bees' health in the field. In this regard, it was reported that the University of Maryland carried out a study in which the complete transcriptome of fat bodies of honey bees fed with the solid product according to the invention (BVP) was analyzed. The observed gene expression profiles were extremely interesting, and the immune genes showed significant improvement under the effect of the solid product according to the invention. However, what most attracted the attention of the researchers was the behavior of the detoxification system. Bees act their own detoxification system to fight exogenous agents such as pesticides, and it results as a side effect that this activation generates reactive oxygen species (ROS), which can damage bees' tissues. Under the effect of BVP, bees expressed fewer of the many genes responsible for the detoxification system, which is a clear indication that BVP helped to detoxify bee systems. One of the ingredients responsible for this effect is [0079] Crocus sativus: the essential extract of Crocus sativus contains crocin, pirocrocin and saffranal, fundamental to increase the resistance of the immune defenses of the bees.
3) Increasing antioxidant power. The powdered dietary supplement product (BVP) not only has 17 to 38 times more antioxidant power than other beekeeping products on the market, but it also proves to be more potent than vitamins B6 and B12 and beta-carotene in combination. This is also due to the presence of the extract of: [0080] Pelargonium graveolens which contains, as noted, geraniol, borneol, citronellol, linalool, terpiheol, limonene, phellandrene, pinene, and methyleugenol, all very active anti-oxidants.
4) Significantly-reducing losses of colonies;
5) Substantially increasing honey production;
6) Improving the overall health status of bees population;
7) Increasing the weight and number of brood per frame;
8) Reducing the losses of queens.

[0081] The following Table 1 shows an exemplary formulation of preferred embodiments of the nutritional and therapeutic preparation for use according to the invention, with preferred concentration ranges for the various ingredients.

TABLE-US-00002 TABLE 1 Preferred formulations PREFERRED CONCENTRATIONS INGREDIENTS (% by weight) Algae 7.0-17.0 Brewer's yeast 0.0-10.0 Glucose, dextrose and/or fructose 50.0-85.0 Acetic acid 0.03-1.5 Glycerin 0.4-0.9 Citric acid 0.01-0.5 Crocus sativus (crocus) 0.05-0.22 Pelargonium graveolens (geranium ess. oil) 0.0-0.10 Geraniol 0.00-0.10 Monarda citriodora (monarda) 0.00-0.18 Melissa officinalis 0.00-0.18 Myristica fragrans (nutmeg) 0.08-0.22 Origanum vulgare (oregano) 0.08-0.22 Origanum majorana (marjoram) 0.08-0.22 Thymol 0.00-0.04 Thymus vulgaris thyme, red and white) 0.05-0.25 Aloe vera or Aloe arborescens 0.10-0.50 Beta vulgaris (common beet) 0.00-0.30 Oxalic acid 0.00-0.10 Fungal mycelium (phylum Basiodimycota) 0.00-1.0

[0082] According to a second aspect complementary to the above, the present invention consists of a method of treatment of colonies of domestic bees for the prevention and the therapy of acariosis through the administration of nutritious and therapeutic substances, which method consists in using the powder product, or in patties, having the following composition: [0083] a) nutritional and tonic ingredients, mainly consisting: of algae containing vegetal proteins, alone or in combination with yeasts; sugars and lower organic acids; [0084] b) natural antioxidants and antiseptics contained in the essential extracts of Origanum vulgare and Pelargonium graveolens or geranium essential oil, and in the essential oils of one or more other aromatic or medicinal plants selected from: Crocus sativus, Monarda citriodora, Melissa officinalis, Myristica fragrans, and Origanum majorana; and [0085] c) substances medicinal to bees, comprising at least one of thymol and essential extracts of Thymus vulgaris, and at least one of oxalic acid, extracts of Aloe vera or Aloe arborescens, geraniol and extracts of Beta vulgaris cv. altissima, and mixtures of two or more of the same.

[0086] In the method of treatment according to the invention, preferred solutions are those wherein the said ingredients are as defined in the dependent claims.

[0087] In current use as a feed for beekeeping, the powder product for use according to the invention can be administered in doses of about 250 ml (1 cup) every 10 days, for a standard hive of 10 frames, administering the product from the top of the frames. When a therapeutic intervention to control the Varroa infestation is necessary, the dose should be increased to 500 ml (two cups) every 10 days, to be administered from the top between the internal frame and the external wall on both sides, or between the frames or directly to the bottom of the hive.

[0088] By way of examples, which are not to be considered as limiting but only as useful to clarify the therapeutic and prophylactic measures for varroa acariosis proposed according to the invention, three exemplary formulations of powdered preparations are reported below, which formulations were tested according to the method of the invention together with the results of some experiments carried out on the application thereof in laboratory and in the field.

EXAMPLE 1

[0089] Nutritional, Antioxidant and Acaricide Formulation with Low Protein Content

[0090] A first exemplary formulation of the product to be used according to the invention is as follows.

TABLE-US-00003 Ingredients used for the production of powder product Ingredients % Dextrose (glucose) 79.7245% Spirulina in powder 10.5000% Dry brewer's yeast 6.3500% Acetic acid, 56% 1.3600% Glicerin 0.7175% Dry extract of Aloe Vera 0.2380% Origanum 50% liq. 0.1625% Essential oil of marjoram 0.1625% Essential oil of nutmeg 0.1625% Dry extract of saffron 0.1625% Essential oil of Melissa officinalis 0.1150% Essential oil white thyme 0.1000% Essential oil of geranium or geraniol 0.0875% Essential oil of red thyme 0.0825% Citric acid 0.0500% Beet powder (Beta vulgaris) 0.0250% Total 100.00%

Process to Produce the Product in Powdered Form

[0091] In order to obtain the preparation, a premix is created, called Premix 1, with the liquid extracts adsorbed in Aerosil 200 (Evonik) and a part of glucose, and another premix, called Premix 2 with the ingredients in powder, i.e. citric acid powder, thymol etc.

[0092] In a horizontal double-helix stainless steel mixer (previously washed and dried) first a half part of the glucose is placed (with the mixer turned off), then the Premix 1 is sieved and then the Premix 2 is also sieved are thus arranged over the previous glucose; then the whole powdered milk and finally the rest of the glucose. The device is operated, mixing for 15 minutes, and immediately thereafter the product undergoes the packaging operations.

[0093] Two small samples representative of the product for the purpose of batch traceability and for quality control are taken at the beginning, in the middle and at the end of the packaging process.

EXAMPLE 2

[0094] Nutritional, Antioxidant and Acaricide Formulation with Spirulina and Yeast at High Protein Value

[0095] A second exemplary formulation of the product to be used according to the invention is as follows.

TABLE-US-00004 Ingredients used for the production of powder product Ingredients % Dextrose (glucose) 60.2245% Spirulina in powder 30.0000% Dry brewer's yeast 6.3500% Acetic acid, 56% 1.3600% Glicerin 0.7175% Dry extract of Aloe Vera 0.2380% Origanum 50% liq. 0.1625% Essential oil of marjoram 0.1625% Essential Oil of nutmeg 0.1625% Dry extract of saffron 0.1625% Essential oil of Melissa officinalis 0.1150% Essential oil white thyme 0.1000% Essential oil of geranium or geraniol 0.0875% Essential oil of red thyme 0.0825% Citric acid 0.0500% Beet powder (Beta vulgaris) 0.0250% Total 100.00%

Production Method

[0096] 1. Water soluble powder at pH 4.3

[0097] 2. Sequence of ingredients [0098] a. Add dextrose and spirulina powders; [0099] b. Add glycerin and acetic acid; [0100] c. Add essential oils individually; [0101] d. Add other ingredients: aloe vera, citric acid, thymol, powder of beet and the other remaining in the list.

[0102] 3. Production roomtemperature and humidity conditions are controlled with air conditioning, to keep temperature around 18 C. (65 F.) and 40-45% relative humidity. The pressure is slightly lower than atmospheric by a ventilation system with filters to reduce dust levels in the air, for the safety of the working environment.

[0103] 4. EquipmentIngredients are weighted with platform scales. A steel horizontal single ribbon mixer with a capacity for 4000 lbs (about 1800 kg) is used. After mixing 3 minutes after last ingredient, the mixer is emptied into an auger hopper, where it goes through a vibrating screen and then it is moved into an overhead bagging bin. A bagging system fills each bag to 22 kg, and each bag is sealed with a heat sealer and placed onto a pallet. Prior to production the labels are applied to empty

EXAMPLE 3

[0104] Nutritional, Antioxidant and Acaricide Formulation with Algae, Yeast and Mushroom

[0105] A third exemplary formulation for producing the product to be used according to the invention is as follows.

TABLE-US-00005 Ingredients used for the production of the powder product Concentrations Ingredients (% by weight) Spirulina, klamath or chlorella in powder 10.50 Brewer's yeast 6.35 Dextrose 79.52 Acetic acid 56% 1.36 Glycerin 0.72 Citric acid 0.05 Crocus sativus (crocus) 0.16 Pelargonium graveolens (geranium essential oil) 0.09 Melissa officinalis 0.12 Myristica fragrans (nutmeg) 0.16 Origanum vulgare (oregano) 0.16 Origanum majorana (marjoram) 0.16 Thymus vulgaris (thyme, red and white) 0.19 Aloe vera 0.24 Beta vulgaris (common beet) 0.030 Fungal mycelium (phylum Basidiomycota) 0.2

[0106] The production method of the final product is the same as the previous example.

EXAMPLE 4

[0107] Nutritional, Antioxidant and Acaricide Formulation with Algae and Yeast to Prepare Patties

[0108] Formula 1 for patties: for maintenance without larvae or eggs in the hive

TABLE-US-00006 Ingredients % Product in powder as per Example 1 66.67% Dextrose syrup 67% 33.33% Total 100.00%

[0109] Formula 2 for patties: for increasing the number of larvae or eggs in hive in brooding period

TABLE-US-00007 Ingredients % Product in powder as per Example 2 62.00% Dextrose syrup 67% 35.00% Vegetal Oil (of palms or coconut) 3.00% Total 100.00%

Production Method

[0110] The dextrose syrup used must be composed of 67% sugar and 33% water. It is preferable that the syrup is glucose (dextrose) or inverted sugar (fractional sucrose). The water to be used must be possible without chlorination or high levels of dissolved carbonates or minerals. When preparing the syrup, avoid heating above 28 C. to avoid the formation of hydroxymethyl furfural as it is poisonous for bees. Mix the dextrose in the water until the solid is completely dissolved.

[0111] After forming the 67% dextrose syrup, add the product of the invention (BeesVita Plus) in powder and mix. In the case of the product of Formula 2 in patties, to increase the number of eggs or larvae during the brooding periods also vegetable oil (palm or coconut) is to be added at the end of the process to obtain a soft dough; not dripping and having a certain malleability. This mixture will represent the final product in patties that can be placed as it is in the hive or wrapped in plastic bags to be cut and used at the time of use or placed on waxed or baking paper in the hive. For periods of long non-use of the product before administration it is possible to sterilize the final mixture with UV rays and use vacuum packs (thus reducing contamination or bacterial attack).

EXPERIMENTAL RESULTS

Cage Tests

[0112] Effect of the Volatiles of the Powder Product According to the Invention (BVP) on Varroa destructor Mortality (Pilot)

[0113] Overview: The pilot study has been carried out to better design an experiment to test the efficiency of BVP volatiles components on Varroa destructor mortality.

Methods: two small 668 inches (15,2415,2420,32 cm) cages were prepared with a bottom chamber to host the BVP powder and keep it separated from the bees. In each cage 18 Varroa infected bees were placed. In the lower chamber of the first cage, separated by a net, 50 g of BVP were placed, while the lower chamber of the second cage remained empty.

[0114] The bees of each cage were provided with a sponge soaked in a 2 M sucrose solution.

[0115] After 48 hours the live bees were harvested and the phoretic mites were counted. Live mites and dead mites of each cage were counted.

Results:

[0116] Initial observations suggest the volatiles of BVP repel mites, as upon being placed in the cage many mites jumped off the bees as compared to the control cage, in which this happened less frequently.

[0117] After 48 hours:

Cage with BVP

TABLE-US-00008 6 bees remained alive 1 phoretic mite 18 fallen mites with 1 alive

Control Cage

[0118]

TABLE-US-00009 7 bees remained alive 5 phoretic mites 6 fallen mites with 2 alive
Discussion with Improvement

[0119] It is evident there is an effect on the drop and mortality of mites that live phoretically on bees. This calls for a more robust experiment with greater statistical power.

Changes in Design:

[0120]

TABLE-US-00010 6 control group 6 groups with BVP 20 infested bees per cage.

Experiments in Field

Field Tests of Anti-Varroa Efficacy

[0121] Three apiaries were tested in the surrounding area of the city of Hamilton, Calif., named Shaw, Gravel East and Gravel West respectively. The study involved a positive control and the tested product, BVP. The positive control consisted of a strip of Mite Away II (NOD Apiary Products USA, Inc.) that was positioned between the two larvae frames. The active ingredient of this product is formic acid. The strips were administered to 34 colonies in each considered apiary.

[0122] The product to be tested, BeesVita Plus, was administered by adding 350 g was administered on 21 Dec. 2017 and finally the third on 28 Dec. 2017. For the apiaries Gravel East and Gravel West the treatment began on 15 Dec. 2017 with the first dose of 350 g, and continued with the second dose on 22 Dec. 2017 and with the third on 29 Dec. 2017.

[0123] In addition, about 150 g of product were also placed on the top of the larvae frames at the second treatment. In each apiary 34 colonies (to be tested by the control body) were treated in this way. Outside the primary study, 59 colonies were treated (strong enough to arrive at the final sampling): 24 at Shaw, 16 at Gravel West and 19 at Gravel East.

Sampling:

[0124] The initial sampling of the study colonies was carried out by the control body on December 13 in Shaw and on December 14 at the Gravel sites. The queen's status was determined either by the presence of eggs or by materially identifying the queen. The bees' frames were taken as a measure of colony's viability and a sample of about 300 bees was taken from each colony and stored in saline to be counted by the University of Maryland control agency to determine the mite levels. The final sampling started on 3 Jan. 2018 and will proceed until completion.

[0125] Measurements of mites taken outside this study were carried out by the Healthy Bees team. the protocol is as follows

[0126] A medium (cluster centric) frame was taken from the hive and inspected for the queen. If detected, it has been removed and placed safely inside the hive. About 300 bees from this frame were placed in a Veto-Pharma Varroa EasyCheck jar (closed by a mesh lid). About 250 ml of 70% isopropyl alcohol were added to the jar, and then the jar was stirred vigorously for about 30 seconds. The fallen mites were then counted, and the container was inspected for any additional mites. After this, the bees were checked to ensure the accuracy of the test.

Results

[0127] The results obtained lead to the conclusion that the BVP product helped control Varroa more efficiently than the common acaricide stripes with formic acid. In terms of the final levels of mites, the colonies treated with formic acid had an average of 4.26 mites per 100 bees and those treated with the product of the invention had an average of 3.26 mites per 100 bees.

[0128] The average reduction of mites per 100 bees with strips with formic acid was 40.35% while the average reduction of mites with BVP was 54.8%.

[0129] Similar field tests were carried out in Turkey in 5 different locations with local bees (Anatolian bee and Caucasian bee), in Brazil (with Africanized bee) and at the IZSExperimental Zooprophylaxis Institute.

Field Tests of Aantivarroa, Aanti-Nosema and Antiviral Efficacy

[0130] In another series of field tests, also conducted in the USA, the effect of the solid product according to the invention, BeesVita Plus (BVP) in the control of varroa infestations, as well as its effect on health of the honey bee colony was investigated.

[0131] A total of 144 colonies from three operations, consisting of 3 apiaries containing 16 apiary colonies (8 controls and 8 treatment colonies) were followed for a period of 2 months. Treatment colonies were fed with BVP every 10 days and were compared with positive control colonies, which received antivarroa treatment with a conventional product appropriate for the region and the time of year. During the whole experimentation, mortality of the colony, incidence of events affecting the queen, size of the population, area of the brood, as well as varroa, nosema and viral loads were measured.

[0132] The objective of the study was to evaluate the effects of the BeesVita Plus product on colony health in field conditions in typical commercial settings, such as a product for spring mite control and feeding. The product was applied according to specifications in half of the colonies, while the other half was subject to the same management practices but without additional feed, with the exception of a standard varroa control treatment.

[0133] It has been noted that, as it happens in the case of any randomized trial conducted under realistic environmental conditions, the initial conditions between bee colonies and between the various apiaries were quite variable. In particular, although bee colonies were randomly assigned to the BVP or to the standard varroa control product groups, those colonies that had been assigned to the BVP treatments showed higher initial varroa rates due to chance, and this pattern remained constant during the study. However, the rate of varroa colonization in groups treated with BVP was not significantly different from the rate of increase in control colonies. Therefore, after taking into account the starting conditions; the evolution of varroa between the colonies treated with BVP and the control colonies, treated with the standard antivarroa product (Apivar and formic acid), was not different.

[0134] This study showed no significant difference in health outcomes between the colonies fed with BVP and the control colonies. Therefore, BVP, as a natural product for the control of varroa, did not behave differently from the two conventional products used as controls in the experimentation. This included no significant difference in the mortality of the colony, in the risk of queen events, in the variation of the population size, in the variation of the brood area, in the variations of the varroa loads, of the nosema loads and in the rates of infection and elimination of 7 target viruses.

[0135] The present invention has been described with particular reference to some embodiments thereof but it should be understood that changes and modifications may be made by persons skilled in the art without departing from the scope of the invention as described in the appended claims.