FIELD OF THE INVENTION

This invention relates to the fields of mosquito maintenance and propagation. More specifically a non-membrane feeding device and diet formulation for mosquito colony production is disclosed.

BACKGROUND OF THE INVENTION

Numerous publications and patent documents, including both published applications and issued patents, are cited throughout the specification in order to describe the state of the art to which this invention pertains. The disclosure of each of these citations is incorporated herein by reference as though set forth in full.

Up to 700 million people are infected and more than a million die each year from mosquito-borne illnesses (Caraballo, Hector (May 2014)). Several diseases transmitted by mosquitoes are malaria, dengue, filariasis, West Nile virus, chikungunya, yellow fever, etc. The next generation of mosquito control technologies seem unlikely to require unilateral use of the toxic chemicals that have been dominate over the past 75 years. The new ecologically benign technologies currently being devised are increasingly focused on genetically modified or sterile mosquitoes, methods that require production of massive numbers of mosquitoes for field release. Female mosquitoes generally require a blood meal to develop eggs for the next generation. Mosquitoes are divergent in host preference, which include humans, animals, birds, reptiles, and amphibians. They also vary in daily rhythms of blood feeding such as diurnal, nocturnal and crepuscular etc. In a laboratory setup, cultures are maintained mostly with a blood feeding system, where female mosquitoes are fed with vertebrate blood. This approach has certain drawbacks. While the diet rich is in protein, diets based on vertebrate blood are expensive, inconsistent in quality, and require pathogen-free certification. Moreover, such blood based diets are unstable during storage. Indeed, the blood must be maintained at 37° C. and most approaches require a membrane to mimic vertebrate skin for the mosquito to pierce. Laboratories maintaining mosquito colonies throughout the world are dependent on live animals or animal blood. Hence, production of large quantities of mosquitoes for mass-release in mosquito control trials places great pressure on animal care facilities and increases maintenance costs several-fold. The blood for these systems usually comes from live animals or birds, which are under constant surveillance and ethical regulations, requiring stringent animal care. Consequently, a laboratory maintaining only a small number of mosquito cages expends thousands of dollars for the blood feeding arrangements which rely on use of animals or artificial membrane feeding system. Understandably then, considerable research effort has been devoted to the development of artificial alternatives to blood. In some labs, researchers use blood components or synthetic blood-free diets based on the host-seeking and blood-feeding behavior of mosquitoes, in which case, the membrane feeding system replaces animal blood or live animals. While an artificial membrane feeding system has been developed and is available commercially (Hemotek), this system still requires the use of collected animal blood. There are huge variations in acceptability of this artificial membrane system for various mosquito species. Blood quality, age and composition also affect system performance (Phasomkusolsil et al., 2013). Often, quality blood for colony maintenance can be difficult to obtain. Some studies have used blood components or synthetic blood-free diet with the membrane feeding system to replace animal blood or live animals. These diets are based on host-seeking and blood feeding behavior of mosquitoes and contain multiple components including phagostimulants, protein sources, digestive activators, buffer solutions, attractants, etc. The performance of these diets depends entirely upon the membrane feeding system. Further, there are concerns regarding long term storage and cost effectiveness of such systems, as adenosine tri-phosphate (ATP), a widely used phagostimulant in the artificial alternatives to blood, used to encourage feeding, is expensive.

Clearly, a need exists for a commercially viable product that will replace costly, labor intensive and ethically vulnerable animal use for blood feeding and artificial membrane feeding systems.

SUMMARY OF THE INVENTION

In accordance with the present invention, an artificial feeding system for maintenance and growth of hematophagous insects is provided which exploits sugar feeding, rather than blood feeding behavior. An exemplary system comprises a diet reservoir for receiving a synthetic, blood-free diet formulation; at least one feeding platform consisting of a mosquito proof covering separating said reservoir from said platform, the system and diet formulation being suitable for egg production and colony maintenance. Another exemplary artificial feeding system comprises a blood-free diet formulation and a feeding device comprising a diet reservoir for receiving the blood-free diet formulation; and a feeding platform for hematophagous insects to sit and feed on the blood-free diet formulation from the diet reservoir. The blood-free diet formulation comprises effective amounts of a protein source and a carbohydrate source, wherein the blood-free diet formulation is suitable for egg production and colony maintenance of hematophagous insects.

In one embodiment, the system comprises a single feeding platform which is optionally a mesh. In another embodiment, the system comprises multiple feeding platforms, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more platforms. In an embodiment, the feeding platform floats over the blood-free diet formulation in the diet reservoir. In another embodiment, at least a portion of the feeding platform is hydrophobic. Yet in another embodiment, the feeding platform is a mesh.

The system can be formed from polypropylene, polyethylene, polystyrene or any other inert plastic material or a metal, such as aluminum foil, or biodegradable materials such as wax paper and the like. The feeding platform can be made of any suitable materials, including but not limited to, polystyrene, aluminum foil, wax paper, sponge, resin, wood and cardboard coated with non-wetting agents.

The diet reservoir of the system can be any shape such as circular or polygonal e.g., square, rectangular or triangular.

In another embodiment of the invention a blood free diet formulation is provided comprising effective amounts of a protein source, a carbohydrate source and, optionally a lipid source. In certain embodiments, the protein source is bovine serum albumin, egg albumin or a mixture thereof. The carbohydrate source can be glucose, sucrose, fructose, or mixtures thereof.

In certain embodiments, the lipid source is cholesterol. In a particularly preferred embodiment, the blood-free diet formulation lacks adenosine tri-phosphate, a phagostimulant and supports several different species of mosquitoes including for example,Aedes, AnophelesandCulexmosquitoes.

In another aspect of the invention, a method of maintaining and growing hematophagus insects is provided. An exemplary method comprises: (a) providing an artificial feeding system as disclosed hereinabove; (b) dissolving the blood-free diet formulation in water; and (c) feeding the hematophagus insects.

