Combination of biological pesticides

Abstract

The present invention relates to a natural pesticide formulation containing: A) botanical products having an arthropod repellent effect (preferably mixture of garlic and chili oils) and B) an entomopathogenic fungus strain or a blend of entomopathogenic fungi strains specific for said arthropod control (preferably blend of Beauveria bassiana strains). The formulation is useful in the control of coffee berry borer (Hypothenemus hampei) in coffee plants and of other important insects in corn crop fields and green-house crop fields (flowers and vegetables).

Claims

1. A pesticide formulation consisting of elements A and B, wherein: A is selected from the group consisting of extracts of: garlic or Allium sativum, and chili pepper, Capsicum sp, Capsicum anuum, Capsicum chinensen, or Capsicum pubescens; and garlic or Allium sativum, and chili pepper, Capsicum sp, Capsicum anuum, Capsicum chinensen, or Capsicum pubescens, and absinthe, Artemisia sp, Artemisia annua, or Artemisia vulgaris; and B is entomopathogenic fungi Beauveria bassiana; wherein the Beauveria bassiana is selected of the group consisting of strains Bb-9001, Bb-9024 and Bb-9119, or blends thereof.

2. The pesticide formulation according to claim 1, wherein the blend of strains has a 210.sup.10 spore/L concentration.

3. The pesticide formulation according to claim 1, wherein A is found in a 0.01% to 20% total weight proportion of the formulation.

4. The pesticide formulation according to claim 3, wherein the blend of strains has a 210.sup.10 spore/L concentration.

5. The pesticide formulation according to claim 1, wherein A is found in a 10% total weight proportion of the formulation.

6. The pesticide formulation according to claim 5, wherein the blend of strains has a 210.sup.10 spore/L concentration.

7. The pesticide formulation according to claim 1, wherein the blend of Beauveria bassiana fungus strains is a blend of Bb-9001, Bb-9024 and Bb-9119 strains.

8. The pesticide formulation according to claim 7, wherein the blend of strains has a 210.sup.10 spore/L concentration.

9. A kit comprising a pesticide formulation consisting of elements A and B, wherein: A is selected from the group consisting of extracts of: garlic or Allium sativum, and chili pepper, Capsicum sp, Capsicum anuum, Capsicum chinensen, or Capsicum pubescens; and garlic or Allium sativum, and chili pepper, Capsicum sp, Capsicum anuum, Capsicum chinensen, or Capsicum pubescens, and absinthe, Artemisia sp, Artemisia annua, or Artemisia vulgaris; and B is entomopathogenic fungi Beauveria bassiana; wherein the Beauveria bassiana is selected of the group consisting of strains Bb-9001, Bb-9024 and Bb-9119, or blends thereof.

Description

3. DESCRIPTION OF THE FIGURE

(1) FIG. 1 (A and B). Experimental unit. 50 fruit branch having coffee berry borer and sprayed with CapsiAlil together with the Beauveria bassiana fungus. A: Absolute control. B: treatment with B. bassiana and CapsiAlil (observe the presence of the white-colored fungus towards the penetration points of the borer in the fruits).

4.BRIEF DESCRIPTION OF THE INVENTION

(2) The present invention provides a natural pesticide formulation for the control of coffee berry borer (Hypothenemus hampei) in coffee plants, and likewise of other insects of economical importance in corn crop fields and greenhouse crop fields (flowers and vegetables), containing A and B elements, wherein A corresponds to botanical products having an insect repellent effect and B consists of an entomopathogenic fungus strain or a blend of entomopathogenic fungi strains specific for arthropod control.

5. DETAILED DESCRIPTION OF THE INVENTION

(3) A natural pesticide formulation was developed for the control of coffee berry borer (Hypothenemus hampei) in coffee plants, and likewise of other insects of economical importance in corn crop fields and greenhouse crop fields (flowers and vegetables), containing A and B elements, wherein A corresponds to botanical products having an insect repellent effect and B consists of an entomopathogenic fungus strain or a blend of entomopathogenic fungi strains specific for arthropod control.

(4) Amongst the entomopathogens, the following species are found: Beauveria sp, Metarhizium sp., Paecilomyces sp., Lecanicillium sp., Nomuraea sp., Isaria sp., Hirsutella sp., Sorosporella, Aspergillus sp., Cordiceps sp., Entomophthora sp., Zoophthora sp., Pandora sp., Entomophaga sp., Conidiobolus and Basidiobolus.

(5) Amongst the botanical products having an insect repellent effect, the following are found: Melia azedarach (Meliaceae): White cedar, a.i. triterpenoids Azadirachta indica (Meliaceae): Nimtree, a.i. Azadirachtin and derivatives thereof (Triterpenoids, limonoids, Nimbina, Salannina). Allium sativum (Alliaceae): garlic. a.i. alliina, allinase, allicine. Capsicum sp. (Solanaceae) including Capsicum annuum, Capsicum frutescens, Capsicum chinense and Capsicum pubescens: chili peppers. a.i. Capsicine. Lonchocarpus utilis (Leguminosae): rotenone. a.i. Rotenone. Chrysantemum cinaerifolium (Compositae): chrysanthemums. a.i. Piretrine and its ester components, formed by the combination of chrysanthemic acid and pyretric acid and the piretrolone, cinerolone and jasmolone alcohols. Nicotiana tabacum (Solanaceae): tobacco. a.i. nicotine. Riania speciosa (Flacourtiaceae): Ryanodine. a.i. Ryanodine. Polygonum hydropiper (Polygonaceae): water pepper. a.i. Polyglodial. Citrus sp. (Rutaceae): lime, citric fruits. a.i. limonoids. Artemisia annua (Asteraceae): sweet wormwood. a.i. Artemisina. Equinacea angustifolia (Asteraceae): Echinacea. a.i. echinicine. Hyssopus officinalis (Lamiaceae): Hyssop. a.i. celo, b-pinene and various monoterpenes such as L-pinocanfene, isopinocanfone and pinocarivon. Lavandula officinalis (Lamiaceae): Lavender. a.i. linalol, eucaliptol, and borneol Mentha pulegium (Lamiaceae): a.i. pulegone, menthol and other terpenic substances such as Menton, isomentone. Ocimun basilicum (Labiatae): Basil. a.i. linalol, estregol, leneol. Artemisia vulgaris, Ambrosia cumanenses (Asteraceae): artemis. a.i. Cielo. Salvia officinalis (Lamiaceae): Salvia. a.i. boreol, cineol, tuyona. Robinia seudoacacia (Fabaceae): false acacia (black locust). a.i. lectin. Rosmarinus officinalis (Lamiaceae): Rosemary. a.i. phenolic acids (cafeic, chlorogenic, rosmarinic), flavonoids (luteol and epigenol derivatives), Diterpenos (carnosol, rosmanol, rosmadial). Tagetes patula (Asteraceae): French marigold. a.i. thiofenes, such as -terthienyl and 5-(3-butene-1-inyl)-2,2-bitienyl (BBT). Melissa officinalis (Lamiaceae): lemon balm. a.i. linalol. Urticasp. (Urticaceae): urtica. a.i. serotonin, histamine, phylosterine Ruta graveolens (Rutaceae): rue. a.i. Rutin, inulin. Canavalia ensiformis (Fabaceae): Canavalia. a.i. Canavalin. Cymbopogon nardos: Citronella. a.i. citronelal and geraniol, I-limonene, canfene, dipentene, citronelol, borneol, nerol, metileugenol. Mentha spicata (Lamiaceae): spearmint. a.i. menthol, phelandrene, centene. Artemisia absinthium (Asteraceae): wormwood, green ginger. a.i. cineol, tuyona. Ocimum basilicum (Lamiaceae): basil. a.i. linalol, estregol, leneol. Calendula otticinalis (Asteraceae): marigold. a.i. calnduline. Minthostachys mollis (Lamiaceae): Mua, Peperina. a.i. Mentol, mentola. Mentha piperita (Lamiaceae): peppermint. a.i. menthol, cielo. Moringa oleifera. Tephrosia purpurea

EXAMPLE 1

(6) The capacity of prompting repellence and mortality on coffee berry borer was tested in field conditions using the combination of the botanical product CapsiAlil, which contains garlic extract (Allium sativum) and chili peppers (capsicum), with the Beauveria bassiana fungus blends. For this, the following treatments were tried: CapsiAlil (at five different concentrations) was combined with the low virulence strains of the Beauveria bassiana fungus (Bb9001, Bb9119, Bb9024), but upon mixing achieve high virulence, at a fungus concentration of 210.sup.10 spores/liter. The botanical product was also evaluated at the same 5 concentrations, the fungus at a 210.sup.10 spores/liter concentration and an absolute control (distilled water) (treatments identified in Table 1).

(7) TABLE-US-00001 TABLE 1 Treatment description for each product. Fungus concentration:CapsiAlil Treatment concentration 1 0:0.03% 2 0:0.3% 3 0:3% 4 0:6% 5 0:10% 6 2 10.sup.10 sp/L:0.03% 7 2 10.sup.10 sp/L:0.3% 8 2 10.sup.10 sp/L:3% 9 2 10.sup.10 sp/L:6% 10 2 10.sup.10 sp/L:10% 11 2 10.sup.10 sp/L:0.03%:0 12 0:0

(8) The treatments were sprayed in parcels holding Coffea arabica Colombian variety 2.5 year old trees, in productive branches holding 120-140 day developed fruits, previously infested with coffee berry borer. The experimental unit was made up of one tree. Each treatment (concentrations and absolute control) comprised 15 experimental units statistically determined variance of 142 associated to the emerging borer average; minimum acceptable difference of 10 borer; greater than 90 reliability and a significance level of 5%.

(9) The experimental units were assigned to the treatments according to the completely random experimental design, and one daily repetition was performed. For each treatment, one branch from the productive zone was randomly taken and all perforated fruits were withdrawn, leaving 50 fruits on the branch. Furthermore, the selected branch was covered using a cylindrical entomological sleeve made from No. 10 caliber wire, 40 cm long and 20 cm diameter, wrapped in white muslin fabric, being fixed to the branch through a polypropylene thread. These sleeves had a window sown in with a Velcro 180 diameter closing along the sleeve in order to visualize the borer emerging behavior. Once the entomological sleeves were installed, infestations in the branch inside the sleeve with 100 borer adults provided by the Biocafe lab were performed. Furthermore, the sleeves were closed with a polypropylene thread and were fastened to the top branch in order to maintain it horizontal. After 24 hours post-infestation, borers were removed, the number of perforations in each infested fruit was recorded and treatment was applied guaranteeing the entire coverage of the branch. For the application, a 1.6 capacity Royal Condor 15-25 pressure accumulation spray unit and a TXVK-3 nozzle (100 cc/min flow at 25 psi) was used.

(10) After each spray, the number of emerging borers from the infested fruits was recorded during 5 minutes of direct observation on each branch. Counting was done through the window.

(11) The emerging percentage was estimated as the result from dividing the number of borers emerged from each branch by the number of total perforations per branch. The window was then closed and the sleeve was left tied to the branch for 20 days.

(12) 20 days after spraying the treatments, the number of adult borers (live and dead) was recorded in each experimental unit. For this to happen, all infested fruits were removed from each branch and deposited in refrigerators inside marked plastic bags in order to be dissected in the lab under a stereoscope. Borer biological status was recorded in each perforated fruit.

(13) Table 2 shows the results of the repellence effect evaluation with regards to the emerging variable caused by the different treatments.

(14) TABLE-US-00002 TABLE 2 Percentage of Borer emergence, 3 days after treatment application. Treatment % emergence EE 1 Capsi 0.03% 5.23 2.2 2 Capsi 0.3% 5.10 2.6 3 Capsi 3% 6.31 1.8 4 Capsi 6% 9.27 2.3 5 Capsi 10% 16.18* 3.3 6 Capsi 0.03% + Bb 6.21 1.8 7 Capsi 0.3% + Bb 5.34 2.1 8 Capsi 3% + Bb 4.85 2.4 9 Capsi 6% + Bb 7.67 3.1 10 Capsi 10% + Bb 15.67* 2.5 11 Bb (relative control) 5.27 2.1 12 Absolute control 5.23 2.3 *Significantly different average with respect to the TA absolute control

(15) The results showed statistical differences between treatments (5% ANOVA -P<0.05). A treatment effect was evidenced when 10% of the botanical product was used, both blended with B. bassiana or alone, according to the comparison test with the absolute control (5% Dunnett test).

(16) Table 3 shows the mortality results on coffee berry borer caused by the different treatment over 20 days. The adult borer mortality obtained in the field after applying the botanical product and the fungus suggest a synergy effect, wherein both the botanical product as well as the fungus inflicted borer mortality of about 45% when applied individually, but when combined, reached a mortality rate in excess of 70%. This evidenced an additive effect causing an additional 30% mortality in coffee berry borer due to the effect of both products.

(17) TABLE-US-00003 TABLE 3 Percentage of dead adult borers, 20 days after applying treatments during the repellence assay. Average % Treatment n mortality EE 1 Capsi 0.03% 40.80 21.07 4.50 2 Capsi 0.3% 37.50 23.74 2.20 3 Capsi 3% 37.30 38.63* 6.65 4 Capsi 6% 39.56 32.36 4.58 5 Capsi 10% 37.44 45.61* 5.91 6 Capsi 0.03% + Bb 37.20 36.62* 7.27 7 Capsi 0.3% + Bb 41.40 42.47* 5.30 8 Capsi 3% + Bb 40.20 51.60* 5.48 9 Capsi 6% + Bb 36.70 50.15* 6.71 10 Capsi 10% + Bb 35.78 74.05* 4.29 11 Bb (relative control) 42.10 43.52 4.54 12 Absolute control 43.50 15.09 2.64 *Significantly different average with respect to the TA absolute control