'Trap to catch mosquitoes." The invention presented here is a kind of trap and method to capture the mosquito of species Aedes aegypti, Aedes albopictus, Anopheles sp and Culex quinquefasciatus in order to monitor, detect and control them. The female Aedes aegypti mosquitoes Linnaeus, 1762 (Diptera: Culicidae) is vector of viral diseases like yellow fever and dengue fever, which spread in epidemic proportions mainly the tropical and subtropical areas of the world, such as West Africa, Central America and South America. The female Aedes albopictus mosquito Linnaeus, 1762 (Diptera: Culicidae) is also vector of viral diseases, such as yellow fever and dengue fever, but on the Asian continent. Recently strains of this species have been found infected with the dengue fever virus.
Mosquitoes of the genus Anopheles are vectors of malaria, with the principal species being Anopheles gambiae, Anopheles darlingi, Anopheles albitarsis, Anopheles aquasalis and Anopheles cruzi.
The Culex quinquefasciatus mosquito is spread throughout Brazil and is a vector of several viruses, most importantly elephantiasis.
The female mosquitoes Ae. aegypti and Ae. albopictus species place their eggs in shaded areas with clear and clean, transparent water, preferably with little organic material. The female mosquitoes of the Anopheles genus place their eggs in larger bodies of water (e.g. lakes, natural pools, etc.) and epiphytal plants, whereas the Culex quinquefasciatus mosquito places its eggs in fetid water containing a large amount of organic material (e.g. standing water, natural pools, lakes, sewers etc.)
The process of searching for and selecting locations for egg-laying by mosquitoes is initiated through endogenous stimuli that determine the start of the activity of flying in females. Later, they are subject to external stimuli, such as, photoperiod, color of the substratum, odor, physical-chemical properties of the water (like p.H.), volume of the breeding place, orifices for egg- laying and vegetation present.
It is known that gravid females are attracted to aquatic environments in order to place their eggs. In general, the waters of these environments contain volatiles proceeding from organic material in decomposition by microorganisms. Female mosquitoes of the Aedes genus are strongly attracted to the color black.
Infusions of organic material associated with water produce volatiles that increase the attraction for several species of mosquitoes to lay eggs (Bentley & Day, Ann.Rev.Entomol.. 34: 401-421, 1989).
Millar et al. (Am.Mosq.Control.Assoc. 8: 11-17, 1992), isolated and identified five compositions from the infusion of fermented "bermuda" grass, which stimulated the egg-laying
of the mosquito Culex quinquefasciatus. The identified compositions are: indole, 3-methyl- indole, phenol, 4-methyl-phenol, 4-ethyl-phenol. Later, see Allan & Kline (J.Chem.Ecol.. 21: 1847-1860, 1995), these compositions were evaluated in ovitraps for Ae. albopictus and Ae. aegypti, in the laboratory and in the field, with the results showing that a great number of eggs were laid by the Ae. albopictus in the water containing the 3-methyl-indole and that the mixture of the five compositions was no more effective than the 3-methyl-indole composition.
U.S. patent # 6.267.953 (2001) Bernier, U.R. et al. for mosquito attractants consist of a compositions of lactic acid in combination with acetone and dimethyl disulfide. This patent also describes the chemical identification of the volatiles of the odor of humans that attract the mosquito in order to feed on the blood of vertebrates.
The Aedes aegypti and Aedes albopictus mosquitoes are controlled by several methods based on the stages of the biological cycle of these insects. The eggs are controlled by means of removal or cleaning of containers that could become egg-laying sites, such as disposables (plastic bottles, plastic cups, etc.) tires, planters. The larvae are controlled by means of chemical larvicides (insecticides) and biological larvicides (entomopathogenic bacteria). The adult mosquitoes are controlled by means of chemical insecticides that are used on the city streets or in strategic locations by means of the technique of ultra-low volume (ULV).
The collection of larvae and pupae of mosquitoes is the most often employed method of monitoring the populations Aedes aegypti and Aedes albopictus in urban areas. This method consists of seeking the immature stages (larvae and pupae) of these species of mosquitoes in potential egg-laying sites (e.g. planters, tires and any container capable of holding water). Once any immature stage is found, the address of the site is registered a sample of the insects found is collected (10 larvae and/or pupas per sample) and transported to the laboratory for analysis of the species. This method is not sensitive, demands a human workforce to find the egg-laying sites in residences or strategic areas of the city, and requires qualified personnel to identify the mosquito in the laboratory by means of a microscope.
The ovitrap, as an alternate method to monitor populations of Aedes aegypti and Aedes albopictus mosquitoes is also used world wide. This trap consist of a black-colored container with approximately a 1 -liter volume and a paddle made of wood or paper with a rough surface as egg-laying substrate. The paddle is attached vertically by a paperclip to the trap. The female Aedes mosquitoes are attracted to the trap and deposit their eggs on the paddle surface. The number of eggs deposited on the paddle is indicative of the population of the Aedes mosquito. Advantages of using this trap are: (a) it is more sensitive method than the collection of immature stages (larvae and pupae); (b) it detects the presence of Aedes in small populations where it would be impossible with the collection method; (c) it is the most effective and low-cost method
and (d) the mosquito searches and deposits eggs in the trap, avoiding the need for monitors to find their own sites.
However, there are disadvantages in the use of this ovitrap, such as, (a) the trap does not capture the females that deposit their eggs there, consequently they can still bite people in order to satisfy their blood source, resulting in the inevitable transmission of dengue fever and yellow fever; (b) the eggs deposited in the trap could hatch, if they should enter into contact with water, becoming a potential growing-site of the Aedes mosquito; necessitating, therefore weekly maintenance checks of the trap. Moreover, (c) it requires a human workforce to count the number of eggs deposited on the paddle (oviposition substrate) in order to determine the level of infestation by the mosquito; (d) it demands the physical space in the laboratory for the development of the larvae; (e) there is a need to identify the larvae species hatched from collected eggs and (f) it requires a period of time greater than 5 days to develop larvae of mosquito collected.
A variation of this ovitrap is described by U.S. patent # 5.123.201 (1992) by Reiter, I.P., which includes an infrared sensor that triggers suction to trap the live, gravid female mosquitoes. When the mosquito crosses the light source, a fan is activated that sucks the mosquito inside the container. This trap is complicated, high cost, and does not kill the captured mosquitoes, thus hmiting its use on a large scale.
Chan, K.L. et. al, "An autocidal ovitrap for the control and possible eradication of Aedes aegypti," Southern Asian Journal of Tropical Medicine and Public Health, 8(1), pages. 56-62 (1977) announce a trap with a modified egg-laying site in which any egg that might hatch would be incapable of emerging as an adult mosquito, due to a mechanical barrier. The authors claim to have been able to reduce the population of the Aedes aegypti mosquito in urban areas by means of egg collection and therefore avoiding the emergence of adult mosquitoes. This model of a trap does not capture the mosquitoes that deposit their eggs; consequently, they can then feed on blood, newly transmitting diseases after which depositing their eggs in other containers.
U.S. patent # 4.328.636 (1982) by Johnson, R.D. describes a trap that allows for the depositing of eggs and development of larvae, but avoids the exit of adult mosquitoes (males and females) that develop inside the trap, by means of a physical screen barrier. This trap does not capture the female mosquitoes that deposit their eggs, so that these egg-laying mosquitoes can then leave the traps and once again bite people, transmitting the dengue fever or yellow fever viruses.
Obaldia, G. Davila, et. al., in "Aedes aegypti resting preference on untreated and deltramethrin-treated crepe paper and plastic foam surfaces", J.Am. Mosq. Control Assoc. 12(3), pages. 467-468 (1996), report research on the use of resting boxes that are saturated with
mosquito insecticide on their surfaces. The limitation of this study as a method for control is that there are so many resting places for mosquitoes to compete with these resting boxes, besides the fact that the females can deposit their eggs before entering into the boxes, guaranteeing a new generation. U.S. patent # 5.983.557 (1999) by Perish et. al. claims a lethal, ovitrap, consisting of the addition of an insecticide to the egg-laying substrate, in order to kill the female Ae. aegypti and Ae. albopictus mosquitoes that land on the substrate to deposit their eggs. This trap does not capture these mosquitoes, which die a certain amount of time after coming into contact with the insecticide. The limitation of this trap is that the mosquitoes can develop a resistance to the insecticide used, if the dose of the insecticide absorbed by the mosquito should be less than a lethal dose and the commtmity may also regret to use traps baited with insecticide in their premisses.
There are other models of traps used to capture adult mosquitoes, with varied functional mechanisms and methods of capturing the insects. U.S. patent # 3.796.001 (1974) by Jackson, S.C. describes a trap that uses an incandescent light to attract mosquitoes, which are subsequently sucked into the interior by means of the flow of air produced by a fan. The fan runs on electric energy, by batteries. The use of this trap is limited for those mosquitoes that are active during the day, and do not respond to a light source, and is also of high cost besides the energy cost for its maintenance. W.O. Patent § 99/35908 (1999) by Wilbanks, A. describes a trap that is capable of attracting mosquitoes by means of varying temperature system, which simulates that of human body temperature, attracting adult mosquitoes. With the help of a fan, they are captured and electrocuted in another compartment. The limitations of this trap are its high cost and the cost of energy needed for its use. U.S. patent # 5.813.166 (1998) by Wigton, B. and Miller, A. describes a trap utihzing the attractants (C02 and octenol) and captures mosquitoes by air flow produced by a fan. The limitations of this trap are its high cost and the cost of energy needed for its use.
Thus, at the present state of the technique, there do not exist traps for the capture of mosquitoes characterized by a combination of a dark container, with at least one opening, with a total or partially sticky inner surface made of paper and/or polymer surface and/or non-limited combinations, where mosquitoes are captured on the internal or external surface.
The invention presented here, is characterized by an ecological trap (eg. it does not use insecticide), simple, low-cost, to capture mosquitoes, as a new alternative method to detect, monitor, and/or control populations of the Aedes aegypti, Aedes albopictus and Culex
mosquitoes in urban areas, thus reducing the rate of transmission of diseases, such as dengue fever, yellow fever, and filariasis (elephantiasis).
The invention presented here is also characterized by an efficient way of capturing the adult Aedes aegypti, Aedes albopictus and Culex quinquefascitus mosquitoes and is composed of:
A geometric format, e.g. cylindrical, rectangle, square and/or an unlimited combination of these formats in unlimited dark colors containing glue on the inside and outside and at least one opening on the end to allow the mosquito to enter;
This trap allows for the capture of mosquitoes on the internal wall of the adhesive structure, thus reducing the number of mosquitoes in the environment, consequently reducing the rate of transmission of disease by them;
Another advantage is the simplified identification of the captured insect, not found elsewhere in the current state of the technique, at the moment of inspection of the trap, therefore avoiding the technical labor and time necessary for identifying the insect in the laboratory; The invention presented here is characterized by the inclusion of even more advantages over conventional methods of monitoring and controlling mosquitoes:
• The trap captures both male and female mosquitoes, thereby reducing the population and mating in the natural environment;
• The female mosquitoes being captured by the trap will not bite any more people as their blood source, reducing the rate of transmission of diseases such as dengue fever, yellow fever, malaria and elephantiasis;
• The strategy of capturing gravid females is superior to the conventional egg-laying trap, this ovitrap allows the females to escape after depositing their eggs and consequently lay eggs in other containers in the environment; • it does not employ insecticides, nor any other toxic agent dangerous to human health;
• the trap can be used without water inside, avoiding the potential of its becoming a site for growing mosquitoes in the future;
• the trap is easily handled, facilitating use by epidemiological security, centers of disease transmission by animals, firms of urban pest control, and by community; • manufacturing the trap is at low-cost and allows for traps to be produced on a large scale, due to the inexpensive raw materials utilized;
• programs for the control of populations of mosquitoes belonging to the genus Aedes, Culex, Anopheles, Mansonia, Wyeomyia, Psorophora, Coquilletidia or insects belonging to the family Psychodidae, genus Lutzomyia.
DETAILED DESCRIPTION OF THE PARTS OF THE TRAP TO CATCH MOSQUITOES The invention presented here can be better understood through the following schematic drawings:
Figure la and 2b is a front view of the invention where the glue can be spread on one of the two sides;
Figure 2 is an example of the several shapes that the trap to catch mosquitoes can take in order to capture mosquitoes: cylindrical (Fig. 2a) or a cut-cone (like a vase) (Fig. 2a), unlimited;
Figure 3 is an overall view of the trap to catch mosquito (Fig 3a and 3b), which can be placed inside the type of egg-laying trap (ovitrap)(Fig. 3c and 3d) commonly used to collect eggs from mosquitoes on paddles. The shape of both traps can be cylindrical or in the form of a cut-cone (Fig 3b and 3d), unlimited;
Figure 4 shows an example of the placement inside the trap of water, natural attractants (e.g. mfusions of organic material) or synthetic mosquito attractants (Fig. 4a) and/or unlimited combinations of these; to capture mosquitoes (Fig.4b);
Figure 5 shows an example of how the trap to catch mosquito (Fig. 5a) can be placed or attached to the inside of a ovitrap (Fig. 5b), commonly used to collect eggs from mosquitoes. Natural attractants (e.g. infusions of organic material) or synthetic mosquito attractants and/or unlimited combinations of these are placed within the ovitraps (Fig. 5c);
Figure 6, shows a sample holder (Fig.6a) for the trap to catch mosquitoes (Fig. 6b) that could be used to avoid contact with the surface of the floor of the place where the trap is located, allowing the range of the attractant to be better distributed in the environment and increase the stability of the trap in the locale. The holder can be made of two unlimited rectangular pieces, which can be located inside the trap (Fig. 6c), in an unlimited way;
Figure 7 illustrates examples of unlimited possible locations for placement of the synthetic attractants in the trap to catch mosquitoes. The attractants can be attached to the inner surface (Fig. 7a) or to the central region (Fig.7b) of the trap to catch mosquitoes, in a unlimited way;
The examples described below illustrate the invention in greater detail, but is not limited to them.
Example 1: Determining the landing site of the insect on the trap: All behavioral tests were carried in the laboratory in acclimatized rooms (27°C, 70% relative humidity and 12h
photoperiod. The Aedes aegypti mosquito was used as the experimental model in all the laboratory experiments. All the tested mosquitoes were gravid females, that were blood fed five days before the experiment. The mosquitoes were let loose in groups, inside transparent acrylic cages (50 x 150 x 50 cm) and the results were obtained within 24 hours after the start of the experiment.
In order to find the first landing site of the gravid female Aedes aegypti mosquito on the egg-laying trap, only one insect was used per test. The experiments were recorded on videotape by means of micro cameras placed 30 cm above the egg-laying trap.
The results shown in Table 1 indicate that, despite not being the egg-depositing place, the surface of the ovitrap was the place that received the greatest number of first landings (60.4%), in comparison to the surface of the water (16.7%) and the paddle (22.9%).
Table 1: Location of first landings of the female Aedes aegypti to deposit their eggs in egg-laying traps. (N=48 repetitions)
Location of first landing of gravid female Total mosquitoes landing (%)
Aedes aegypti mosquitoes
Surface of the water 08 (16.7)
Paddle 11 (22.9)
Internal wall 29 (60.4)
Example 2: Optimizing the placement of the glue: Having determined that the location in which the females landed more frequently was the internal wall of the trap and the paddle (oviposition substrate), an odorless glue commonly used to capture insects was placed, but not limited to, these locations, with the objective of confirming the best location to capture the adult insect. The bioassay methodology used was the same as in example 1. The results shown in Table 2 indicate that the location of collection of the greatest number of mosquitoes was the inner surface of the container. Therefore, the inner surface was the location selected for the placement of the glue to capture mosquitoes.
Table 2: Choice of location in the ovitrap for the placement of glue (N= 67 Aedes aegypti females). Total of 3 repetitions.
Location of glue in the trap Total mosquitoes captured (%)
Control (without glue) 0 (0.0) paddle 10 (14.9)
Internal wall 36 (53.7)
Example 3: Comparison of the method of glue application by hand with the "hot- melt" method of PVC cards: Having defined the location to place the glue, polymer cards (eg.
PCN) were covered with glue by hand, or alternatively with a "Hot-Melt" method of applying hot glue that melts. The results in table 3 show that glue applied by hot melt application is more efficient than the application by hand.
Table 3: Percentage of gravid Aedes aegypti females captured in the laboratory on the inside of the ovitraps, containing hand-applied glue versus industrially produced sticky cards.
Repetitions Application by hand "hot-melt" application
(% of captured females) (% of captured females)
1 95.4 95.4
2 100.0 100.0
3 100.0 100.0
4 66.7 95.6
5 81.2 82.3
6 71.4 73.7
Average ± s.d. 85.8 ± 15.64 91.2 ± 10.75
Example 4: Effect of the size of the trap on the capture of mosquitoes. In the laboratory, the effect of the size of the trap was evaluated to the capture of adult Aedes aegypti mosquitoes. Two sizes were used, the normal (10 cm diam. x 12 cm) and a reduced (5 cm diam. x 7 cm). It can be observed in Table 4 that the size of the trap influences the number of adult Aedes aegypti captured, in that the normal size captured 55.4%, whereas the reduced size trap captured only 30.4% of the mosquitoes.
Table 4: Effect of the size of the ovitraps on the capture of Aedes aegypti mosquitoes in the laboratory.
9 22 0 0.0 12 54.5 10 45.5 10 17 2 11.8 15 88.2 1 5.9
Total 204 27 13.2 113 55.4 62 30.4
Average 26.6 2.5 10.0 13.4 50.8 9.7 39.6
Standard Deviation - 2.00 - 2.50 - 3.19 -
Example 5: Effect of the shiny or matte surface on the external part of the trap to catch mosquitoes. This experiment aimed to study the effect of light reflection on the outside of the trap on the capture of Aedes aegypti mosquitoes. A trap with light reflection on the external part of the trap was compared to one with a non-reflecting external area. The results in Table 5 show that mate black outer surface is more effective to catch mosquito than shiny outer surface.
Table 5: Effect of light reflection on the external part of the trap versus non-reflective surface on the capture of gravid female Aedes aegypti mosquitoes.
No.
Repetition females/test Shiny black trap (%) Matte black trap (%)
1 10 2 (20.0) 6 (60.0)
2 10 2 (20.0) 5 (50.0)
3 10 1 (10.0) 9 (90.0)
4 10 4 (40.0) 6 (60.0)
Total 40 09 - 26 -
Average - 2.25 (22.5) 6.50 (65.0)
Example 6: Effect of the shape of trap on the capture of mosquitoes. Usually, the ovitrap used to capture the eggs of the Aedes aegypti or Aedes albopictus mosquito is made in a cut cone format (like a vase-shaped). Therefore, two types of shapes for the trap were analyzed: a cone and a cylinder. The cylindrical shape was chosen because it reduces the amount of light striking the inner surface of the trap and the glue contained there. The results show that the cylindrical shape (62.2%) caught a greater number of gravid females Aedes aegypti mosquitoes than the cone shape (32.4%). Table 6: Influence of the shape of the trap on the capture of gravid female Aedes aegypti mosquitoes.
Number females/ Females not Cylindrical shape cut cone
Repetition test captured (Vase-shaped) n Total (%) Total (%) Total (%)
1 25 4 16.0 13 52.0 5 20.0
2 31 2 6.5 14 45.2 12 38.7
3 23 5 21.7 8 34.8 6 26.1
4 18 0 0.0 11 61.1 4 22.2
5 19 2 10.5 10 52.6 5 26.3
6 21 6 28.6 9 42.9 4 19.0
7 31 4 12.9 13 41.9 8 25.8
8 18 2 11.1 8 44.4 7 38.9
9 26 0 0.0 12 46.2 10 38.5
10 20 2 10.0 15 75.0 1 5.0
Total 232 27 11.6 113 48.7 62 26.7
Average 23.2 2.7 13.3 11.3 62.2 6.2 32.4
Standard
Deviation 4.93 2.00 - 2.50 - 3.19 -
Example 7: Effect of remove water or natural attractant (grass infusion) from the trap to catch mosquito in the field.
This experiment was conducted in the field. For each repetition, a pair of traps (one containing water and other without) was placed 1 meter apart. The minimum distance between each repetition was at least 100 meters. The results in Table 7 show that there was capture of
Aedes aegypti in both traps, however, the presence of water caught higher number of mosquitoes.
Table 7: Results of the capture of Aedes aegypti mosquito in field traps with and without water.
Ovitrap with Repetition Total sticky board Rl R2 R3 R4
With water 2 3 3 (5 8* without water 1 3 1 0 5 Example 8: Comparison between the sensitivity of the trap to catch mosquitoes with egg-laying trap (ovitrap): the trap to catch mosquitoes collects insects stuck to the inner surface of the trap whereas the egg-laying trap collects eggs deposited on its paddle. The objective of the present test was to evaluate in an urban area (field test) the sensitivity of both traps. The results in Table 8 show that the trap to catch mosquitoes collected a total of 19 mosquitoes in 15 repetitions (an average of 1.27 insects/test). The table 9 shows the positivity for ovitrap and trap to catch mosquito as presence or absence of eggs and adult, respectively. The frequency of mosquito caught in the trap to catch mosquitoes was 100% positive. In contrast, the egg-laying trap collected eggs only through three repetitions, obtaining 20% of
positive traps. Therefore, the sensitivity of the trap for the capture of mosquitoes is greater than that of egg-laying. This trap for capturing mosquitoes also showed efficiency in the capture of, particularly, female Aedes aegypti mosquitoes.
Table 8: Comparison of sensitivity between the egg-laying trap and the trap for the capturing of adult mosquitoes
Trap to catch mosquitos Egg-laying
Ae. aegypti Ae. albopictus trap
Repetition Total (Number of
Male Female Male Female eggs)
1 0 1 0 0 1 0
2 0 1 0 0 1 0
3 0 1 0 0 1 0
4 0 1 0 0 1 0
5 0 0 0 0 0 15
6 0 0 0 1 1 0
7 1 1 0 0 2 0
8 0 3 0 0 3 0
9 0 0 0 1 1 35
10 0 1 0 0 1 79
11 0 1 0 0 1 0
12 0 2 0 0 2 0
13 0 2 0 0 2 0
14 0 1 0 0 1 0
15 0 1 0 0 1 0
Total 1 16 0 2 19 129
Average 0.07 1.07 0.00 0.13 1.27 8.60
Table 9: Index of positivity for the trap for capturing mosquitoes and the egg-laying trap. (+)= presence of eggs or adult mosquito (-) absence of egg or adult mosquito.
Example 9: The effect of the addition of synthetic attractant on the trap to catch mosquitoes in the laboratory. The synthetic oviposition attractant was added to the trap and tested in the laboratory in order to evaluate the increase in the capture of the mosquito. Four groups of 10 gravid female mosquitoes each of Aedes aegypti were released in cages (50 x 150 x 50 cm), five days after having fed on blood. Two traps were placed inside the cages, 1 meter apart, one containing only water and the other containing 50 ul of oviposition attractant (nonanal) in 300 ml of water. The number of mosquitoes captured was observed after 24 hours. The results show that the addition of the synthetic oviposition attractant increased the total number of mosquitoes captured (Table 10).
Table 10: Effect of the addition of synthetic oviposition attractants to the trap for capturing mosquitoes in laboratory.
Example 10: The effect of the addition of synthetic attractants on the trap to catch mosquitoes in the field. Each synthetic oviposition attractants were added individually to the trap to catch mosquitoes and tested in the field. Two traps for capturing mosquitoes were placed two meters apart in shielded area in urban area. One was baited with synthetic oviposition attractant and other with grass infusion as control. The number of mosquitoes captured was observed after 24 hours. The results show that the addition of the synthetic oviposition attractant nonanal and decanal increased the total number of mosquitoes captured when compared with grass infusions (natural attractant)(Table 11).
Table 11: Effect of the addition of synthetic attractants such as decanal and nonanal to the trap for capturing mosquitoes in the field.
Example 11: The effect' of combination of synthetic attractants on the trap to catch mosquitoes in the field. Combinations of the synthetic oviposition attractants were used in order to enhance capture of the mosquito. Two traps for capturing mosquitoes were placed two meters apart in shielded area in urban area. One was baited with combination of synthetic oviposition attractant and other with grass infusion as control. The number of mosquitoes captured was observed after 24 hours. The results show that the combination the synthetic oviposition attractant such as decanal and nonanal with p-cresol increased the total number of mosquitoes captured (Table 12).
Table 12: Effect of the addition of synthetic attractants such as nonanal and decanal in combination with p-cresol to the trap for capturing mosquitoes in the field.
Repetition Infusion p-cresol + decanal Infusion p-cresol + nonanal
1 1 1 1 1
2 1 1 1 1
3 1 1 3 1
4 1 1 1 1
5 0 2 0 0
6 0 1 0 0
7 0 1 0 0
Total 4 8 6 4