Methods and Apparatus for Luring and Sterilizing Male Mosquitoes
Posted on August 12th, 2016
Note: This text would was used in a patent application filed in early April, 2016, but the Patent and Trademark Office has let us down. Lobbyists and Congress weakened patent protections with the America Invents Act, and patent trolls from big business simply litigate with individual inventors until they get what they want1. At this writing, the PTO averages 16.1 months before it takes the first action on a patent application, and 25.7 months to process the average application. Over 500,000 patent applications are now awaiting examination2.
The present invention lures male mosquitoes of a target species with a sound generator that mimics the sound of a flying female of that species. Males are drawn to the sound and find a tray of honey mixed with chemosterilant, which they ingest. Because a male mosquito mates multiple times, and a female mates only once, sterilizing the males will efficiently reduce the population. The invention is contained in a box with small perforations that admit mosquitoes but exclude larger insects. The inner walls of the box provide an additional landing surface and are coated with chemosterilant. The lure sound is intermittent, and when turned off, sterilized males who have entered the box and eaten are expected to go forth and mate.
The field of the invention is Sterile Insect Technology (SIT), more particularly a means of selectively sterilizing male mosquitoes in the field.
U.S. Classification: 43/124: Devices and processes the primary object of which is to destroy or kill vermin without trapping them.
CPC Classification: A01M 31/008 Lure dispensing devices. And A01M 1/023 Stationary means for “non-chemical sterilization” of invertebrates
Mosquitoes of the genus Aedes are known to transmit dangerous arboviruses (arthropod-borne viruses) that infect humans and other invertebrates. Aedes aegypti and Aedes albopictus are the principal vectors of dengue (DENV-1, DENV-2, DENV-3, DENV-4), chikungunya (CHIKV), yellow fever (YFV), and Zika (ZIKV) viruses (CDC, 2016). Others in this genus, such as Aedes africanus are also suspected of carrying these viruses and transmitting them to humans. (Haddow et al, 1964).
Together, A. aegypti and A. albopictus are now distributed throughout the tropical, subtropical, and temperate world. In the U.S., their combined range extends from Florida to California and north through Illinois and New York. Both are highly invasive species, whose range is expanding rapidly. A albopictus has expanded its geographical range from Southeast Asia and India to include North and South America, Europe, Africa and the Pacific region in the past three decades (van de Hurka, et al, 2016). Both species choose to live in close proximity to people.
A. aegypti has been shown to feed on humans, dogs, cats, horses, and chickens (Barrera et al, 2012) , guineapigs, rats (Gouck, 1972), and songbirds such as Cardinals (Richards et al, 2006) Both species are anthropophilic – they prefer humans as their hosts (Tandon and Ray, 2000).
Controlling the population of these mosquitoes is critical to controlling the spread of the viruses they transmit.
Devices for attracting and destroying insects are well known in the art. For example, U.S. Pat. No. 4,908,979 to Hostetter discloses a device that uses a sex attractant, a third generation pesticide, and a black light attached to streetlights to primarily to attract Gypsy Moths. U.S. Pat. No. 4,519,776 to DeYoreo et. al., U.S. Pat. No. 4,907,366 to Balfour, U.S. Pat. Nos. 5,657,576 and 6,088,949 to Nicosia, U.S. Pat. Nos. 5,799,436 and 6,055,766 to Nolan et al., U.S. Pat. No. 6,050,025 to Wilbanks and U.S. Pat. No. 6,305,122 to Iwao all disclose devices that use a scent attractant and a form of heat to lure mosquitoes. U.S. Pat. No. 5,255,468 to Sheshire teaches a device that uses light, heat and motion as an attractant. U.S. Pat. No. 5,280,684 discloses killing insects and specifically flies in trash cans using garbage as a scent lure. U.S. Pat. No. 5,369,909 to Murphy teaches modifying an electric fence on a farm for the control of flies. U.S. Pat. No. 4,907,366 to Balfour uses lactic acid and water heated to simulate human breath. All such devices seem to focus on immediately killing flying insects, rather than sterilizing them, and are generally indiscriminate in the insects they will kill.
Popular Control Methods
Numerous approaches to controlling mosquitoes are now in use:
- Insecticides may be used to spray spaces, using backpacks, trucks, or aircraft. While these are effective, they often kill more than the intended species, and may be toxic to humans. Mosquitoes develop resistance to insecticides and pass this resistance on to their young. Furthermore, depending on the application method and climatic factors, only 10% of conventionally applied insecticides may reach their target in sufficient time and quantities.(Kenawy, 1998)
- Insecticides have succeeded in the form of lethal tire piles consisting of 7–9 water-filled tires treated with λ-cyhalothrin and s-methoprene pellets, to which female A. albopictus are attracted and come into contact with the insecticide.3
- Screens and air conditioners help keep mosquitoes outdoors, protecting those who stay inside. This doesn’t fully solve the problem, for not everyone can afford screens or air conditioners, and sooner or later everyone has to venture out.
- Repellents may be applied to clothing and to skin, and often deter mosquitoes. But if the repellent is not applied to some part body part, such as the back of an ankle, a mosquito may find a suitable meal.
- Mosquito traps, such as the DCD Autocidal Gravid Ovitrap and the AGO trap, which lure gravid females and then capture them with sticky glue when they try to lay eggs has shown to be effective in reducing populations of A. aegypti. Others use pheromones as a lure (Ong and Jaal, 2015) Some traps are likely effective but are definitely expensive. The BG-Mosquitaire CO2 costs about $275 US, and requires an electrical connection and a tank of CO2 to operate. The BG-Mosquitaire costs about $175, requires electricity and the addition of a $20 mix of artificial skin emanations every two months.
- Eliminating containers that the mosquitoes lay their eggs in would be a good idea, and can be locally effective. But it would require collecting all glass, metal, and plastic containers, collecting used tires, replacing septic tanks with sewer systems, removing bird baths and flower pots, and the like. The abundance of such containers in the environment makes this a daunting task.
- Chemical larvicides, such as temephos, can be added to containers where larvae may be found. But such larvicides may accumulate in the environment, and may be indiscriminate in what they kill. Given the abundance of suitable containers for breeding, adding chemical larvicides to all of them is more daunting that picking the containers up and removing them from the environment.
- Biological larvicides, including bacteria and enzymes, likely have little impact on non-target species and don’t accumulate in the environment. But again, adding them to all of the suitable breeding containers available is a daunting and expensive task.
- Biological control with aquatic predators is effective for many species of mosquitoes, but not for A. aegypti and A. albopictus, which often develop in small containers that may completely dry out between rains.
The methods have not eliminated the spread of A. aegypti and A. albopictus. In fact, their range is spreading, reflecting their growing adaptation to living with humans, and perhaps in response to a warming planet. A better method may be needed to supplement the various control methods now in place.
Novel Control Methods
Some recent experiments in controlling mosquitoes should be highlighted. All show promise, but none of these are yet beyond the experimental stage:
- A bait (not a trap) is loaded with male-killing bacteria and an attractant.4
- Bait is loaded with bacteria that protects against some disease. Disease dies, disease-free mosquitoes prosper. For example, Wolbachia (bacteria) can protect the mosquito from dengue.5 Wolbachia is a genus of bacteria that can live within the cells of anywhere from 20-70% of the world’s insects. In mosquitoes, males and females infected with Wolbachia must possess the same strain to successfully reproduce — if there’s a mismatch, the bacteria will sabotage the male’s sperm, effectively sterilizing him. In an egg fertilized by sperm with an incompatible Wolbachia strain, the male chromosomes never form properly, and the embryo dies. MosquitoMate is a firm that is developing a product called “ZAP Males”, which are male mosquitoes that the company has infected, by injection, with a strain of Wolbachia different than the one normally associated with the species. If Wolbachia could be introduced to a male mosquito through ingestion, then a tainted bait could be provided, but a method of doing this has not yet been developed.
- Release males that don’t produce the hormone that helps the ladies lay eggs. Every time they mate, they knock out a lady, since the ladies only have sex once.6
- Trap the ladies. The BG GAT is a trap that needs no power, no CO2. Kills with canola oil. Can use water as an attractant. Might work without an LLIN – Long Lasting Insecticide-Treated Net. 7
- Irradiate males, sterilize them and then release them. They’ll mate several times, each time knocking a female out. 8
Because none of the traditional methods of controlling mosquito populations have been sufficiently effective, and because none of the novel control methods have moved beyond the experimental stage, there remains a considerable need for a method and device that can control mosquito populations.
Any success in controlling mosquito populations requires an understanding of their reproductive behavior.
Mosquitoes may mate when they have been adults for at least 2 days.
Mating typically occurs in the first hour of darkness. Males approach females, using the female flight tone of their species to guide them. Females who have mated previously actively resist being clasped by a male.
After injecting semen, seminal secretions produced by the male accessory glands (MAGs) are transferred to females in the form of a coagulated mass called the mating plug. This internal mating plug allows the female to store the sperm – without it, sperm are not stored correctly. The plug also prevents any future male from inserting sperm. The MAG secretion transferred during insemination induces long-term sexual refractoriness in females, but this only takes effect 2 to 3 days after insemination (Helinski, 2012).
Males lack the piercing mouthparts needed for biting. They are able to get enough energy for their daily needs from flower nectar. Females of many species of mosquitoes need a meal of blood for the protein needed by their egg yolks. Some research has shown that some strains of A. aegypti, under some conditions, are autogenous, and if there are nutritional reserves remaining from the larval stage, may use those reserves to produce their first eggs, seeking blood meals only when those reserves are depleted. But A. aegypti appears to normally be anautogenous – always requiring a blood meal prior to egg laying.
Only female mosquitoes bite, obtaining a blood meal to nourish the eggs they will lay. When a female bites, any viruses in her saliva may be transferred to the bird or mammal she has bitten, and the blood she has taken may contain viruses from that bird or mammal.
Over its lifetime, one sterile male might fully inseminate a maximum of 7 females and might transfer a partial amount of semen to a further maximum of 8 females. Over the same period of time, untreated males could fully inseminate up to 11 females and partially inseminate another 9 females. (Oliva et al, 2013)9
Brief Summary of the Invention
The present invention provides a method and device for reducing a mosquito population through the field sterilization of male mosquitoes. As may be seen in the figure, it appears to be a small box that can be hung, or placed on any horizontal surface. The box has small holes that allow a mosquito to enter and exit, but that prevent larger insects, such as butterflies and bees, from entering. At the bottom of the box is the lure: a sound generator which includes a small battery and speaker. Above the lure at the bottom of the box is a tray of the chemosterilant mixed with honey or other food preferred by male mosquitoes. This tray is covered with a screen that will give the mosquitoes a means of landing just above the honey, without getting stuck in it. The remaining surfaces inside the box are coated with the chemosterilant, which will be expected to contribute to the sterilization through contact. Such surfaces will have enough texture to give a mosquito a foothold while they are in the box.
The present invention is not a mosquito trap, but more like a feeding station. Males are lured by the sound that a female makes in flight. The sound emanates from beneath a source of food preferred by male mosquitoes such as honey, that food laced with an odorless chemosterilant. The sides and top of this station are coated with a chemosterilant, which begins to do its work when a male lands. Males position themselves on a screen just above the chemosterilant-laced honey and eat their fill. When the simulated sound of a flying female has been turned off, they exit the station and head off in search of a female to mate with.
With this invention, male mosquitoes are sterilized in situ, where they occur, not requiring that they be raised in a laboratory and released. It uses normal, native mosquitoes that have not been genetically engineered. It does not require the use of pesticides, produces no side effects on humans or other species, and produces no damage to the ecosystem.
The present invention is species-specific in its sterilization: male mosquitoes are drawn to the sound of females of their species in flight.
Sterilized males do the work of finding females before they have been inseminated, no matter where they may be hiding, and whose insemination prevents them from laying viable eggs. Once inseminated, female mosquitoes will not mate again. Unlike the other applications of the “Sterile Insect Technique”, this invention will produce an endless supply of local sterile males, and not require laboratory labor to raise, sterilize, and distribute sterilized males.
This invention is intended to be manufactured at very low cost so that it may be distributed to every household in a region concerned about Zika or battling mosquitoes.
Brief Description of the Drawing
Fig. 1 Vertical cross-sectional view of one embodiment of the invention. It is a small box that can be hung, or placed on any horizontal surface. At the top of one embodiment is a handle (1). The box has small holes on one or more sides (2) that allow a mosquito to enter and exit, but that prevent larger insects, such as butterflies and bees, from entering. At the bottom of the inside of the box is the lure (3): a sound generator which includes a small battery and speaker. Above the lure at the bottom of the box is a tray of the chemosterilant mixed with honey or other food preferred by male mosquitoes (4). This tray is covered with a screen (5) that will give the mosquitoes a means of landing just above the honey, without getting stuck in it. The remaining surfaces inside the box are coated with the chemosterilant, which will be expected to contribute to the sterilization through contact. Such surfaces will have enough texture to give a mosquito a foothold while they are in the box.
Detailed Description of the Invention
Using Mosquitoes to Control Mosquitoes
Female mosquitoes will only mate once in their lives, but can lay eggs several times, biting each time. Males mate repeatedly, and as noted, do not bite.
Our approach is to attract male mosquitoes with a lure, feed them, sterilize them, and release them. In their remaining days on earth, they will mate repeatedly, but never fertilize a female. Each female partner will never mate again, will not develop eggs, and will not bite a human or any other animal. As a result, she will not contract any viruses through her biting, and will not spread any.
Mosquitoes “hear” with their Johnston’s organ, at the base of their antennae, which detects air particles displaced by vibrating wings. Flying mosquitoes generate sound that is sex-specific and species-specific. Males are able to use such sound to determine if a passing mosquito is a female or male, and if it is the same species as the male. Our invention will lure males with a sound that would be made by females of the species of interest, such as A. aegypti.
The search to define the distinctive sounds for A. aegypti has been ongoing since at least 1948.
- Roth (1948) first showed that A. Aegypti males responded to sounds between 100 and 800 Hz by attempting copulation.
- This range was narrowed by Nijhout & Craig (1971), who found that male A. aegypti, A. formosus, A. mascarensis , A. albopictus, A. polynesiensis, and A. scutellaris responded to tones between 400 and 500 Hz.
- The range was narrowed still further by Ikeshoji(1981) who found that A. aegypti males responded best to sounds of 466 Hz and that A. Albopictus males responded best to sounds of 462 Hz.
- Johnson and Ritchie (2015) did well luring A. aegypti males with a synthesized pure tone of 484 Hz.
The use of female flying sounds as a lure for males is not expected to work equally well with all species of mosquito. A. aegypti uses sound in courtship, whereas it appears that A. albopictus does not.
- Ikeshoji (1985) found that sound attracted A. albopictus males but did not elicit copulatory behavior as it does in A. aegypti males.
- Duhrkopf and Hartberg (1992) found evidence that the role of sound is more important in the mating of A. aegypti than in the mating of A. albopictus. A. albopictus males did not respond to recordings of either A. albopictus or A. aegypti females, but A. aegypti males responded preferentially to the recorded sounds of A. aegypti females.
In one embodiment of our invention, the lure for males of a mosquito species will use a recording of the flight of a female of that species. In another embodiment, the lure will be a synthesized tone at the frequency that males of the target species find most attractive. We anticipate that a recording of a female might be somewhat more effective at attracting males, but that the generation of a tone at some single fundamental frequency, such as 465 Hz, might be cheaper to produce.
The best amplitude of the sound will depend on ambient noise, the extent to which the lure muffles sounds emanating from it. In one embodiment, an earbud speaker would produce a sound pressure level of 57.8 dB at 2 cm distance.
The audible lure will be timed to turn off and back on. When the sound is turned off, we expect the disappointed males to eat the chemosterilant until they are satiated and then fly off. When the sound is turned on, any males in the area would be lured. The intervals can be determined in the field, but perhaps 5 minutes on, and 5 minutes off would work. A satiated male, full of chemosterilant but still in the area, might happen into the trap a second time, but would not be expected to ingest more sterilant on this visit.
The Sterile Insect Technique (SIT) is a method of biological control in which sterile insects are released into the wild. When sterilized males mate, the females produce no offspring. The technique was used successfully to eradicate the screw-worm fly from much of North America, as well as eradicating the Mexican fruit fly, the Tsetse fly, the Medfly, and the Melon fly from parts of the world.
The general idea of sterilizing male mosquitoes is not new, but previous implementations have had their problems.
- SIT normally relies on the release of large numbers of sterile insects. Mass rearing facilities initially produce equal numbers of both species, and before release, the females and males must be separated and the females “discarded”. Genetic techniques for removing females from the release population have been developed for several insect species using conventional mutagenesis methods, but not yet for mosquitoes. Researchers at the Imperial College London are working on a technique of raising mosquitoes with a female-specific dominant lethal gene, an advancement in “RIDL” – Release of Insects carrying a Dominant Lethal gene. In this approach, genetically modified mosquitoes would be raised in the lab, and then released. Most of the females with the gene would soon die; the males would not need to be sterilized, and so would be healthy upon release, when they would mate with mosquitoes not raised in the lab, passing on the lethal gene. RIDL is still experimental. One solution, proposed here, is unattended field sterilization.
- Fertile A. aegypti eggs can survive months of desiccation, so any success that might come from a one-time release of sterile males soon wanes as new mosquitoes hatch from old eggs. The solution to this is to release sterile males non-stop, whenever males are found.
- Sterilization is traditionally done with gamma rays or x-rays, termed radiosterilization. It is the goal to set radiation levels to kill a male’s sperm, but not so high as to kill or injure the male himself. In fact, radiation usually leaves the male weakened, and with a shortened life span, making him less able to compete with non-irradiated males. And it is not practical to include radioactive material in a cheap lure that will be used in a populated area. One solution to this is to use other approaches, such as chemosterilization, that target sterility without otherwise damaging the individual.
- Releases of sterilized males have not always reduced the population of mosquitoes. But such conclusions might be different if the number released significantly changed the proportion of wild sterile males. When essentially all males are sterile, fertility must drop, and the population must begin to disappear.
SIT works best when females of the targeted species only mate once, which happens to be the case with mosquitoes.
Our invention will sterilize male mosquitoes by providing a mix of a chemosterilant such as apholate or tepa mixed with a carrier such as honey. The chemosterilant will be diluted to a level which proves most effective in causing sterility without otherwise harming life span or reproductive behavior.
In one embodiment, tepa (AKA aphoxide, methyl aphoxide, and metepa) is used as the chemosterilant. Tepa is a colorless and odorless liquid chemosterilant. It is highly soluble in organic solvents such as acetone, ether, and alcohol.
- Tepa can inactivate sperm in mosquitoes and other insects (housefly, Mexican fruit fly, German cockroach, and citrus red mite.)10
- Weidhaas et al. (1961) obtained complete sterility in A. aegypti when adults were fed a honey solution containing 0.1% apholate or 0.5% tepa (aphoxide).
- Weidhaas (1962) spread tepa on glass at 10 mgm. per sq. ft., and confined male mosquitoes to the surface for 4 hours. Females that mated with these males were able to lay eggs (in 92% of the 5 females studied), but no eggs hatched. Another test showed that males could be effectively sterilized in the fourth day of life.
- Bertram (1963) obtained complete sterility for a period of 15 days in males exposed for 3 hours on deposits of 200 mg of thiotepa per square meter on paper surfaces.
- Dame & Schmidt (1964) obtained 96% sterility with 1% of metepa on honey.
In one embodiment, a mix of honey and 0.5% tepa might be used as the chemosterilant, and provided to mosquitoes for ingestion. In another embodiment, the lure is simply placed in a small box, whose insides are coated with tepa. The male mosquito, attracted to the lure, walks in the tepa and is sterilized. Our preferred embodiment provides both methods of distributing tepa to a visiting mosquito.
Tepa is economical. Online through Alibaba, it sells for as little as $45 per kilogram – enough to treat 100,000 square feet. But there are many other chemosterilants that might be used in an embodiment.
- A portable male mosquito lure and sterilizer for use in suppressing mosquito populations comprising a box with:
- a lure: a sound generator on the bottom which periodically generates the sound of a flying female of the species of interest,
- a tray positioned above the sound generator, containing a chemosterilant mixed with a carrier: a food source preferred by male mosquitoes, such as honey,
- a screen covering the tray of chemosterilant, to allow a landing mosquito to eat without getting stuck in the carrier-chemosterilant mix,
- sides and lid of the box coated on the inside with chemosterilant (without honey), and providing a landing surface for mosquitoes,
- holes in the sides and lid of a size to permit mosquitoes to enter and exit, but not so large as to admit insects such as butterflies or bees,
- a hinged box top, to allow for inspection, replacement of the chemosterilant, or replacement of the battery used by the sound generator,
- a handle on the top of the box.
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- Eden, Scott. “The Greatest American Invention”. Popular Mechanics, July/August 2016 p. 93-99 ↩
- Patents Data at a Glance. USPTO. ↩
- van den Hurka et al, 2016 ↩
- See popular story: https://www.washingtonpost.com/news/speaking-of-science/wp/2016/03/11/could-feminist-bacteria-help-fight-off-the-mosquitoes-that-carry-zika/ and http://onlinelibrary.wiley.com/doi/10.1111/1365-2664.12638/abstract and http://onlinelibrary.wiley.com/doi/10.1111/1365-2664.12638/pdf and get the PDF for $38. It isn’t even published yet. ↩
- https://www.washingtonpost.com/national/health-science/field-tests-show-bacterial-oddball-can-be-a-dengue-destroyer/2011/08/24/gIQA7qBobJ_story.html ↩
- http://www.scientificamerican.com/article/new-insight-into-mosquito-sex-could-aid-in-conquering-malaria/ ↩
- See http://www.pacificbiologics.com.au/images/files/manuals/BG_GAT_manual.pdf and See http://www.biogents.com/cms/website.php?id=/en/traps/biogents-trap-systems/bg_gat.htm ↩
- See http://www.dw.com/en/messing-with-the-sex-life-of-the-mosquito/a-17725668 ↩
- Sources for this section: Charlwood and Jones (2008), Gwadz (1969), Hagedorn & Judson (1972), Klowden (1981), Rogers et al (2009), Trpis (1977) ↩
- Kalyani et al(2015). ↩