WO2013051953A2

WO2013051953A2 - Use of essential oils to manufacture mite controlling product in honey bees

Info

Publication number: WO2013051953A2
Application number: PCT/PL2012/000099
Authority: WO
Inventors: Jerzy KAZIMIERCZAK; Wiesław LONDZIN; Danuta BOMBIŃSKA; Sylwia ΡΟRĘΒSΚΑ; Krzysztof ŻELECHOWSKI
Original assignee: Instytut Przemyslu Organicznego
Current assignee: Instytut Przemyslu Organicznego
Priority date: 2011-10-06
Filing date: 2012-10-05
Publication date: 2013-04-11
Anticipated expiration: 2014-04-06
Also published as: WO2013051953A3

Abstract

This invention is related to application of tea tree oil and/or basil oil to produce the preparation to control mites in honey bee family, particularly control Varroa destructor or Typhlodromus pyri Sch.

Description

Use of essential oils to manufacture mite controlling product in honey bees

This invention is related to application of essential oils, single or in mixture, to manufacture mite controlling product in honey bees.

Mites occurring in human environment as well as other mites occurring in natural environment transmit pathogenic microorganisms responsible for many dermatoses and systemic diseases. Varroa destructor is a distinct group of mites attacking insects, particularly useful insects. It is particularly dangerous for honey bees because it brings about varroatosis, a threatening disease destroying bee colonies.

So far, control of mites in bees consists mainly in application of pesticides. As examples of the known agents used against mites one can mention fipronil, amitraz, fluvalinate, coumaphos, bromfenvinphos, and the others. These preparations were applied to protect household animals against ticks as well as in treatment of varroatosis in bees. However, the chemical preparations should be used with an exceptional care due to their high toxicity.

In recent years many preparations of natural origin, such as thymol, menthol, oxalic acid, formic acid, have been used for treatment of bees. Polish patent description No. PL 186883 describes a slow-release gel preparation designed for i. a. mite control. Natural oils could be the active substance in this preparation.

The inventors brought attention to the preparations of natural origin, such as natural essential oils, particularly those applied for centuries in natural medicine. In the Polish patent applications No. P.390096 and P.390098 use of fir oil was proposed (main ingredients are pinenes and limonene) and rosemary oil (main ingredients are eucalyptol and pinenes) to control mites in bees.

Essential oils are liquid, volatile, aromatic substances occurring usually in plant special cells of secretory tissue. Currently ca. two thousand species of essential oil plants occurring in all climatic zones are known. As far as chemistry is concerned, oils are unusually complex mixtures of up to a few hundred different chemical compounds which content is not completely known and often variable. Therefore unequivocal confirmation of real biological activity of particular essential oils or their mixture is difficult. As a consequence it is not possible to draw conclusion about usefulness of particular essential oils based on applications of the other oils. It is known in the prior art that oil from a tea tree obtained from Melaleuca alternifolia Cheel plant belonging to Myntaceae plant family is applied for a long time in folk medicine, in bacterial and viral infections. The main ingredient of a fresh tea tree oil is terpinen-4-ol. Its content varies in the range of 29-45%. Similar compounds γ-terpinen, a- terpinen, a-terpineol, and monoterpenes occurr in slightly smaller quantities. Basil oil (Ocimum basilicwn L.) is also known. Its main ingredients are methylchavicol, linalol, eugenol, and citral.

From the patent descriptions US 2002022043 and DE202006012918 is known application of tee tree oil and basil oil to control mites in house dust present in residential rooms as well as in textile materials, such as bed linen and fitted carpets. The German patent description No. DE 19824683 discloses application of tea tree oil to sprinkle surfaces and air to control mites in house dust. In the German patent description No. DE 19824680 an essentials oil, e. g. tea tree oil, was applied to control household dust mites after depositing it on activated charcoal granules. Application of tea tree oil mixed with the potassium soap and the other essential oils to clean skin from mites and lice was presented in the Australian patent description No. AU 2008101219. According to the German patent description No. DE 19824681 tea tree oil was applied to remove mites and lice from the skin and hair. The Korean patent description No. of publication KR20030026067 presents composition to control household dust mites which ingredients contain i. a. basil oil. This oil functions in the composition as an attractant while an insecticide is responsible for the mite control.

However, the solutions presented in the above patent descriptions refer exclusively to the dust mites feeding mainly on epidermis. Yet mites constitute a very large order of arachnida having ca. 30 thousand genera of diversified structure, including saprophags, parasites (mainly ectoparasites), and predators. Furthermore, mites are adapted to life in different environments what results in diversified species features. Their common feature is only fusion of head, thorax, and abdomen as well as lack of external segmentation. In consequence, reactions of particular mites to influence of the same factors cannot be predicted. It concerns particularly parasitic mites adapted to coexistence with different organisms.

The known publications lack any suggestion of possible application of tea tree oil or basil oil to control particular type of mites attacking bees. The essence of the invention is use of tea tree oil or basil oil to produce preparation controlling mites in bees, particularly Varroa destructor or Typhlodromus pyri Sch. Mites.

Preferably tea tree oil or mixture of tea tree oil with basil oil is applied in concentration of 0.05 to 0.5 g/dm³, compared to a beehive capacity.

Preferably basil oil is applied in concentration of 0.1 to 0.5 g/dm³ compared to a beehive capacity.

Preferably tea tree oil and/or basil oil is applied in a mixture with the other essential oil or its component and/or with a chemical miticide, such as fipronil, fluvalinate, coumaphos, bromfenvinphos, or amitraz. Preferably the mixture of tea tree oil and/or basil oil is applied with rosemary oil, pepper mint oil, fir oil, and/or thymol. Preferably tea tree oil, basil oil or their mixture is applied alternately with the chemical agents controlling mites, such as fipronil, fluvalinate, coumaphos, bromfenvinphos or amitraz.

Preferably tea tree oil and/or basil oil are put into the beehive making use of a molded article moistened with the oil or a molded article covered with the oil. The molded article can be made from a natural material or plastics. Preferably a molded profile covered with oil containing a microlayer is applied.

It is also preferred when the oil is directly applied to pouring out or beehive spraying, and basil oil is preferably applied as vapours.

The effectiveness of tea tree oil and basil oil to control mites was examined with two bioindicators: predatory mite Typhlodromus pyri Sch. and Varroa destructor.

Examination with Varroa destructor was carried out in test cages on bees with parasiting mites on them. The cages were placed in tight containers to which wicks saturated with the tested oil were introduced. Based on a wick mass loss a dose of the oil was calculated which got through to the cage. Activity of the oil was assessed based on amount of dead mites. When the test was finished formic acid was put to the wick and the fallen mites were assessed after this treatment. The test showed a high activity of tea tree oil and basil oil. The mites were completely eliminated (100% activity) with concentrations below 0.5 g/dm . Tests with application of the mixture of tea tree and basil oil with thymol and the mixture with the other essential oil - rosemary oil, peppermint oil, and fir oil - showed equally high activity.

Typhlodromus pyri Sch. mites closed in a tight test equipment were exposed to the oil vapours. The assessment of miticidal activity of the tested substances consisted in determining percentage of mortality and atypical behaviour of mites. The tests showed a high activity of tea tree oil and basil oil towards the predatory mite - mortality up to 100%.

Tea tree oil and basil oil or their mixture can provide an excellent supplement or can substitute the chemical agents for honey bee protection. Particularly good results have been obtained applying only the oil or mixture with thymol, peppermint oil or fir oil. It is also possible to apply an alternative therapy, e. g. application in one year one of the chemotherapeutics, such as amitraz, and oils in the next year. It will result in decrease of threat caused by e. g. accumulation of active products or decomposition products in honey or in wax. It is possible also to apply a strategy of a safer oils application during small or medium escalation of varroatosis, and only in cases of a very strong threat to use chemotherapeutics.

Below activity of tea tree oil will be illustrated in the examples.

Example 1

Bees together with the mites parasiting on them were placed in test cages and transferred to the test equipment. Next wicks of 24 cm surface and volume of 8 cm moistened with tea tree oil, tea tree oil mixed with basil oil and thymol were placed for 24 h in the test equipment. 1 ml of tea tree oil or 0.5 ml of tea tree oil were put on every wick and on the second wick 0.5 ml of other tested oil was put (when a combined effect was examined). The wicks moistened with oils were weighed before installation in the test equipment and again after removal. Based on this mass of the oil (dose) evaporated during the 24 h exposition was calculated. When the application was over, i. e. after 24 h, the dropped, dead mites were counted. Then an additional operation with 60% formic acid and 0.25 ml/wick dose was carried out and after the consecutive 24 h the dead mites were counted again. The effectiveness of the tested oil was calculated according to the formula:

S /,. =— _s - 100%

Where:

SE - effectiveness of the procedure,

E - number of dropped mites under influence of the oil,

S - total number of mites eliminated by the tested oil and formic acid.

During testing no adverse effect on the bees was observed. Results in the control of Varroa destructor are shown in the Table 1. Table 1

Number of dropped off mites A - after application of the given substances (individuals) after 24 h)

Number of dropped off mites B - after application of the reference substance (formic acid) (individuals) after 24 h.

Example 2

Activity of the basil oil was tested according to the method described in the example 1. Results in the control of Varroa destructor with basil oil application are shown in the Table 2,

Table 2

0.1 26 9 35 74.3 79.0

Basil oil 23 7 30 76.7

28 0 28 100.0

0.2 26 0 26 100.0 100.0

Basil oil 29 0 29 100.0

27 0 27 100.0

0.3 21 0 21 100.0 100.0 basil oil 33 0 33 100.0

0.1 basil oil 34 0 34 100.0 100.0

+ 0.1

thymol

0.1 basil oil 41 2 43 95.3 95.3 + 0.2

pepper

mint oil

0.1 basil oil 39 1 40 97.5 97.5 + 0.1 reference 2 18 20 10.0

0.0

1 28 29 3.4 8.5

3 22 25 12.0

Example 3

The model equipment consisted of the glass tight container having capacity of 1 dm³ holding three test stands. A basic set comprises plates (3 cm x 3 cm x 0.5 cm) with a central opening (1.5 cm), glass slides (3 cm x 3 cm) as well as appropriate squares of absorbent paper. Such sets were assembled in the chambers by means of neutral glue as well as rubber belts. The test equipment designed in such a way enables a gas contact of mites with vapours of the essential oils and an easy observation of the effects of their action.

Cubes (2 cm x 2 cm x 2 cm) cut from a flonstic sponge were moistened with the essential oil (2 ml) and weighed before placing in the container. After 24 h the sponge was weighed again to calculate the amount of the evaporated oil.

In the test the essential oil was assessed with three repetitions, with 5 individuals in every test, in total 15 individuals. Simultaneously with the tested series a reference experiment was carried out (3 x 5 mite individuals), without any contact with the tested material.

Food in form of the common pine (Pinus sylvestris) pollen was put into the test chambers followed by adult individuals Typhlodromus pyri Sch taken from the breeding ground and inserted with a delicate brush. The chambers were placed vertically on the glass stands in the tight containers to which floristic sponge moistened with a specific essential oil was also inserted. The mites were exposed to essential oil vapours for 24 h.

The experiment was carried out in the controlled laboratory conditions required for the test organisms. Temperature and humidity monitoring was recorded continuously with a thermohygrograph.

Mortality of the mites was assessed after 24 h, counting from the time, when mites were transferred to the test equipment.

Mortality observations in the tested groups were referred appropriately to the reference group and recorded in the chart with readouts of results.

Stereoscopic microscope was used in the observations. Using a clean small brush the mites were classified in the following categories:

Dead mites - when they did not react to the triple stimulation and were immobile;

Mites in a state of agony - when they still twitched but were unable to straighten up;

Living mites - when they moved spontaneously without showing any sign of paralysis; Affected mites - when they showed signs of diminished coordination in comparison to the mites from the reference group.

Mortality of the mites was calculated adding dead, missing, and individuals in the state of agony. This value was compared to the number of mites occurring in the beginning of the experiment. The missing individuals are mites which could not be found in the test area. The results are shown in the Table 3.

Table 3.

Tea tree oil and basil oil show high activity (mite mortality up to 100%) towards the predatory mite Typhlodromus pyri Sch. Their high volatility allows to obtain higher concentrations in the air assuring by the same token the high effectiveness.

Claims

1. Use of tea tree oil and/or basil oil to produce mite controlling preparation for bees.

2. Use according to claim 1, characterized in that the preparation is applied to control Varroa destructor or Typhlodromus pyri Sch mites.

3. Use according to claim 1 , characterized in that tea tree oil alone or in a mixture with basil oil is applied in concentration from 0.05 to 0.5 g/dm³ compared to the capacity of the beehive.

4. Use according to claim 1 , characterized in that basil oil is applied in concentration from 0.1 to 0.5 g/dm³ compared to the capacity of the beehive.

5. Use according to claim 1 or 2, characterized in that tea tree oil and/or basil oil is applied in a mixture with the other essential oil and/or chemical miticide, such as fipronil, fluvalinate, coumaphos, bromfenvinphos, amitraz.

6. Use according to claim 5, characterized in that the mixture of tea tree oil and basil oil with rosemary oil, pepper mint oil, fir oil, and/or thymol is applied.

7. Use according to claim 1, characterized in that tea tree oil and/or basil oil is applied alternately with the chemical miticides, such as fipronil, fluvalinate, coumaphos, bromfenvinphos, amitraz.

8. Use according to claim 1 , characterized in that tea tree oil and/or basil oil is put to the beehive making use of the molded article moistened with an oil or of the molded article covered with an oil.

9. Use according to claim 8, characterized in that the molded article is made of the natural material or plastic.

10. Use according to claim 8, characterized in that the molded article covered with a microlayer containing oil is applied.

1 1. Use according to claim 1, characterized in that tea tree oil and/or basil oil is applied to pouring out or beehive spraying.

12. Use according to claim 1, characterized in that basil oil is applied as vapours.