Statement of Problem
The condo complex where I live has a huge mosquito problem not only in the monsoon season but also in the prior dry, hot season in which large amounts of water is used to keep non-native plants alive. Attempts have been made to keep the mosquitoes under control by use of Bacillus thuringeniesis spores. I question whether there is enough organic matter in the top soil for these spores to really propagate. Like many places in the Tucson/Phoenix area, the condo complex has a large number of china berry trees, Melia azedarach. The gardeners do not seem to be very motivated to clean up the fruit and leaves that fall to the ground.
The fruit and leaves could be rapidly dried on asphalt in most regions where Melia azedarach are common. The died plant material could be mulched, and mixed with Bacillus thuringeniesis spores.
These china berries were collected, chopped in a blender with some tap water, and incubated in a warm window. Some Bacillus subtillus natto spores were added to the endogenous bacteria.
Quick Test with support from Sterility Assurance Labs?
- Prepare a semi-dry mulch of Melia azedarach leaves and/or fruit.
- set aside some of the mulch for later extraction
- Sterilize the mulch to kill endogenous microorganisms.
- Test the ability of the mulch to support growth of Bacillus thuringeniesis spores.
- Extract the fermented mulch with additional water and/or some other solvent mix.
- Extract the semi-dry mulch set aside in step 2 n the same manner as step 5. Do the bacilli help liberate the mosquito killing compounds?
- Compare profile using usual analytical means: hplc, UV/Vis absorbance, TLC, and so on.
- Contact the U of A Dept of Entomology. Someone there, perhaps a grad student, might be willing to help with assays out of the reach of U of A IP rights?
Other groups have used a 50% ethanol extract. Perhaps just water would be the more relevant solvent. If digested long enough, chopped Chinaberries turn to a mulch.
What is unique
Melia azedarach extracts are very popular means of mosquito controls in parts of the developing world. It recently came to my attention that in addition to containing a lot of cellulose, plants also contain beta-glucans. The Bacillus genus has the capability of producing beta glucanase enzymes. The bacteria degrade the yard waste. The yard waste propagates the mosquito killing bacteria.
Whirl wind tour of prior art
Melia azedarach fruit samples were collected from the campus of Akdeniz University.
- Fruit was dried at 25 °C about 2 weeks and ground to fine powder using blender.
- The powder was extracted with methanol for 2 days at 25 °C.
- Undissolved debris was removed with a Whatman No.1 filter paper.
- The extract was dried under vacuum rotary evaporator.
- The Ulex pipiens mosquito was used in the studies. U pipiens is found in Arapsuyu, Antalya, Turkey
The EC50 was only 170 ppm. The LC90 was 1099 ppm. (Koc 2016). Without reading the paper, I’m not sure how they calculated parts per million. All we might want to know is if bacteria and Chinaberry mulch synergize in killing mosquitoes.
example 2, nanoparticles
Another study tested Melia azedarach fruit extracts formulated into silver nanoparticles. These were tested in
- soil and water
- larvae and pupae (Anbu 2016).
Ramanibai and Velayutham (2016) fractionated a leaf extract and tested a fraction that contained a pure compound.
- A. aegypti LC50 = 7.94, LC990= 23.82 ppm
- C. quinquefasciatus (and LC50 = 13.58 andLC90 = 40.03 ppm).
example 3, a mosquito repellent rather than a larvacide.
An extract of Melia azedarach is marketed in Africa as a mosquito repellent. It is more effective than no treatment but not as effective as DEET in an informal study (Abyi 2015). Leaf burning has also been reported as a mosquito repellent in parts of Africa (Mavundza 2011)
example 5, seed extracts
Trude and Bomblies (2011) conducted a study on Anopheles arabiensis in the village of Asendabo, Ethiopia.
- Locals harvested the fruit and separated the pulp from the seeds.
- The seeds were powdered.
- Chinaberry seed powder was added to the larval containers at three treatment levels: 5 g m-2, 10 g m-2 and 20 g m-2, with 100 individual larvae in each treatment level and a control.
- The containers were monitored daily and larvae, pupae and adult mosquitoes were counted.
Wandscheer (2004) ethanol extracted seeds.
- They reported lethality to Aedes aegypti in terms of gram %.
- They commented that the extraction could be more efficient if co-extracted natural fats were also removed. Removing the pulp is too much work. I think the pulp can be used as a growth substrate for Bacillus thuringenesis. Let an anti-insect bacterium do the work of removing the pulp! As for the co-extracted fats, these might also be subject to bacterial degradation.
example 6, leaf extracts
Prophiro (2008) used fresh and dried leaves
- Extracted with various v/v mixtures of ethanol and water.
- Ethanol extracts more effective at killing the Aedes aegypti larvae.
Coria (2008) compared leaf and fruit extracts on killing the same species of larvae.
- The leaf extract was shown to be more effective in this study.
- Coria (2008) did not separate the pulp from the seeds.
Isman (2006) mentioned fermentation products in the abstract.
- We may want to get this paper.
Nathan (2006) used methanol to prepare extracts of leaves and seeds from the chinaberry tree.
- used the Anopheles stephensi mosquito.
- The extracts were testing on all aspects of mosquito life from larvae to breeding and biting adults.
- The seed extracts showed high bioactivity at all doses, while the leaf extracts proved to be active, only in the higher dose.
Some of these references claim that chinaberry leaf and fruit extracts are cheaper in US dollars than traditional pesticides. This may be true in places in the developing world where labor is cheap.
aclm/(Melia and azedarach) This is a search of the US Patent database for all claims that cite the genus species name of china berry. These are fun reading if you have an interest in sustainability.
Patent 10 might have some overlap except that the are looking at
What is claimed is:
1. A pesticidal composition, comprising: a non-aqueous, water miscible solvent at least 20% by weight; two or more plant alkaloids selected from the group consisting of toosendanin, tomatine, stemonine, nicotine, anabasine, matrine, oxymatrine, sophocarpine, N-oxysophocarpine, cytisine, and aloperine, wherein the composition is formulated to function as a pesticide.
2. The composition of claim 1, wherein the solvent is selected from the group consisting of triacetin, polyethylene glycols, polysorbates, poloxamers, polyoxyethylene ethers, polyoxyethylated alkylphenols, copolymer of alkylphenol epoxyethane and epoxypropane, N-methyl pyrrolidone, propylene glycol, ethyl acetate, dimethyl sulfoxide, dimethyl acetamide, benzyl alcohol, 2-pyrrolidone, benzyl benzoate, C.sub.2-6 alkanols, 2-ethoxyethanol, 2-ethoxyethyl acetate, methyl acetate, ethyl acetate, ethylene glycol diethyl ether, ethylene glycol dimethyl ether, and tributyrin.
3. The composition of claim 1, wherein the solvent is polyoxyethylene ether.
4. The composition of claim 1, wherein the solvent is polyoxyethylated alkylphenol.
5. The composition of claim 1, further comprising an extract derived from a plant selected from the group consisting of Anabasis aphylla, Nicotiana acuminata, Duboisia myoporoides, Zinnia elegans, and Zollikoferia eliquiensis, which contains anabasine.
6. The composition of claim 1, further comprising an extract derived from a plant that selected from the group consisting of Melia toosendan Sieb. et Zucc. and Melia azedarach L., which contains toosendanin.
Abiy E, Gebre-Michael T, Balkew M, Medhin G. (2015) Repellent efficacy of DEET, MyggA, neem (Azedirachta indica) oil and chinaberry (Melia azedarach) oil against Anopheles arabiensis, the principal malaria vector in Ethiopia. Malar J. 2015 14:187. doi: 10.1186/s12936-015-0705-4.
Anbu P, Murugan K, Madhiyazhagan P, Dinesh D, Subramaniam J, Panneerselvam C, Suresh U, Alarfaj AA, Munusamy MA, Higuchi A, Hwang JS, Kumar S, Nicoletti M, Benelli G.(2016) Green-synthesised nanoparticles from Melia azedarach seeds and the cyclopoid crustacean Cyclops vernalis: an eco-friendly route to control the malaria vector Anopheles stephensi? Nat Prod Res. 30(18):2077-84. doi: 10.1080/14786419.2015.1114935.
Coria C, Almiron W, Valladares G, Carpinella C, Ludueña F, Defago M, Palacios S. (2008) Larvicide and oviposition deterrent effects of fruit and leaf extracts from Melia azedarach L. on Aedes aegypti (L.) (Diptera: Culicidae). Bioresour Technol. 99(8):3066-70. Epub 2007 Jul 31.
Isman MB.(2006) Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Annu Rev Entomol.51:45-66. Review.
Koc S, Evren OH, Cetin H. (2016) Evaluation of Some Plant Fruit Extracts for the Control of West Nile Virus Vector Culex pipiens (Diptera: Culicidae). J Arthropod Borne Dis. 10(4):595-601. eCollection 2016 Dec.
Mavundza EJ, Maharaj R, Finnie JF, Kabera G, Van Staden J. (2011) An ethnobotanical survey of mosquito repellent plants in uMkhanyakude district, KwaZulu-Natal province, South Africa. J Ethnopharmacol. 137(3):1516-20. doi: 10.1016/j.jep.2011.08.040.
Nathan SS, Savitha G, George DK, Narmadha A, Suganya L, Chung PG.(2006) Efficacy of Melia azedarach L. extract on the malarial vector Anopheles stephensi Liston (Diptera: Culicidae). Bioresour Technol. 97(11):1316-23. Epub 2005 Jul 27.
Prophiro JS, Rossi JC, Pedroso MF, Kanis LA, Silva OS. (2008)Leaf extracts of Melia azedarach Linnaeus (Sapindales: Meliaceae) act as larvicide against Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae) Rev Soc Bras Med Trop.41(6):560-4.
Ramanibai R, Velayutham K.(2015) Bioactive compound synthesis of Ag nanoparticles from leaves of Melia azedarach and its control for mosquito larvae. Res Vet Sci. 98:82-8. doi: 10.1016/j.rvsc.2014.11.009. Epub 2014 Nov 20.
Trudel RE, Bomblies A. (2011) Larvicidal effects of Chinaberry (Melia azederach) powder on Anopheles arabiensis in Ethiopia. Parasit Vectors. 4:72. doi: 10.1186/1756-3305-4-72.
Wandscheer CB, Duque JE, da Silva MA, Fukuyama Y, Wohlke JL, Adelmann J, Fontana JD. (2004) Larvicidal action of ethanolic extracts from fruit endocarps of Melia azedarach and Azadirachta indica against the dengue mosquito Aedes aegypti. Toxicon. 44(8):829-35.