There are several types of viral pathogens that cause disease in mosquitoes with most belonging to 4 major groups. The most common viruses of mosquitoes are the baculoviruses (NPVs) (Baculoviridae: Nucleopolyhedrovirus) and cytoplasmic polyhedrosis viruses (CPVs) (Reoviridae: Cypovirus). The other major types of viruses in mosquitoes are represented by the densoviruses (DNVs) (Parvoviridae: Brevidensovirus) and the iridoviruses (MIVs) (Iridoviridae: Chloriridovirus). Baculoviruses, densoviruses and iridoviruses are DNA viruses while cypoviruses are the main RNA viruses in mosquitoes. This chapter presents an overview of the recent advancements in the study of mosquito pathogenic viruses and discusses how this new understanding of virus-mosquito interactions can be used to develop novel research and control strategies.

Lagenidium giganteum is a facultative parasite of mosquito larvae that initiates infection by production of biflagellate zoospores that selectively recognize and attach to larval cuticle. Following penetration of the cuticle, the parasite proliferates within the host, killing it within 24–60 h. Under optimum conditions the mycelia differentiate to produce asexual and/or sexual reproductive structures that produce zoospores within hours (asexual stage) to amplify the initial infection, or remain dormant for days, months or years (sexual stage), until conditions are conducive to mosquito breeding and spore germination. Recycling following a single application has been documented for up to 8–10 years. Environmental conditions that reduce or eliminate zoospore production, including temperature extremes (less than 16°C or greater than 32°C) and moderate levels of salinity and organic load, preclude use of the parasite for operational mosquito control. Three formulations of L. giganteum have been registered with the USEPA. Widespread use of the parasite will be possible when yields of the sexual stage in liquid culture are increased by a factor of ca. 10².

Cyclopoid copepods have proved more effective for practical mosquito control than any other invertebrate predator of mosquito larvae. Their operational potential is enhanced by the fact that mass production is relatively easy and inexpensive. The exceptional potential of copepods for mosquito control was first realized about 25 years ago. Since then, laboratory experiments with copepods and mosquito larvae around the world have shown:

Most field testing of copepods has been in Aedes container-breeding habitats. Field tests have shown that:

Field surveys in Anopheles, floodwater Aedes, and Culex breeding habitats have shown that natural copepod populations can substantially reduce, or even eliminate, mosquito production. Field trials in temporary pools, marshes, and rice fields have demonstrated that introduction of the right copepod species to the right habitat at the right time can eliminate Anopheles or floodwater Aedes larvae. As a rule, copepods cannot eliminate Culex production by themselves, but they can reinforce and augment control by other methods.

The only large-scale operational use of copepods to date has been in Vietnam, which has achieved local eradication of Ae. aegypti in hundreds of villages. Conditions in Vietnam are particularly favorable because:

Copepods have the potential for local eradication of Ae. aegypti and Ae. albopictus in many other countries besides Vietnam. Professional capacity for copepod management and social institutions for community participation to help with implementation and maintenance are the main factors limiting broader use of copepods for operational mosquito control at the present time.

Toxorhynchites is an unusual and interesting genus of large, non-biting mosquitoes. In spite of their size, they are – like many species of mosquitoes – completely harmless to man. The larvae are predaceous on other mosquitoes and aquatic organisms that inhabit both natural and artificial containers. Because this habitat is the source of several medically important species of mosquitoes, there is warrant for evaluating the potential of Toxorhynchites as a biological control agent under various conditions. Toxorhynchites is not seen as a panacea for the control of all container-inhabiting mosquitoes. However, it has demonstrated practical potential in certain restricted but important situations.

Since the discovery of Bacillus thuringiensis (Berliner) serovariety israelensis de Barjac (Bti) and efficacious isolates of Bacillus sphaericus Neide, formulations of these bacteria have become the predominant non-chemical means employed for control of mosquito larvae at several locations in the United States and other countries. An overview of developments in the past 20 years is presented in this chapter regarding the toxins of Bti and B. sphaericus, their modes of action, efficacy and factors that affect larvicidal activity, development of resistance, safety, and their roles in integrated mosquito control. The efficacy of Bti formulations has been demonstrated in a variety of habitats against a multitude of species of mosquitoes. B. sphaericus formulations have been utilized predominantly in organically enriched habitats against Culex species, but they are also active in a variety of habitats having low organic enrichment, against numerous species, and across several genera. Stegomyia spp. are not susceptible to practical doses of B. sphaericus formulations. B. sphaericus has been shown to persist longer than Bti in polluted habitats and, under certain circumstances, can recycle in larval cadavers. A disadvantage of B. sphaericus has been the development of resistance in certain populations of Cx. quinquefasciatus Say and Cx. pipiens Linnaeus. Biotic and abiotic factors that influence the larvicidal activity of Bti and B. sphaericus include species of mosquito and their respective feeding strategies, rate of ingestion, age and density of larvae, habitat factors (temperature, solar radiation, depth of water, turbidity, tannin and organic content, presence of vegetation, etc.), formulation factors (type of formulation, toxin content, how effectively the material reaches the target, and settling rate), storage conditions, production factors, means of application and frequency of treatments. Due to their efficacy and relative specificity, both Bti and B. sphaericus can be ideal control agents in integrated programs especially where other biological control agents, environmental management, personal protection and the judicious use of insecticides are combined.

Genetic engineering techniques have been used to significantly improve mosquito larvicides based on the bacteria Bacillus thuringiensis (Bt) subsp. israelensis (Bti) and Bacillus sphaericus (Bs). These new larvicides hold excellent promise for providing better and more cost-effective control of nuisance mosquitoes and vectors of important diseases, including the anopheline vectors of malaria and culicine vectors responsible for filariasis and viral encephalitides. The toxicity of Bti and Bs is due primarily to endotoxin proteins produced during sporulation. After ingestion by larvae, these are activated and destroy the larval stomach, quickly resulting in death. By cloning the genes encoding various endotoxins from Bt and Bs species, and engineering these for high levels of synthesis, we have been able to generate recombinant bacterial strains based on Bti that are more than 10 times as effective as the conventional strains of Bti or Bs that serve as the active ingredients of commercial bacterial larvicides currently used for mosquito control. The best of these recombinants contain all major Bti endotoxins, specifically, Cry4A, Cry4B, Cry11A, and Cyt1A, plus the binary (Bin) endotoxin of Bs, the principal mosquitocidal protein responsible for the activity of this species. The presence of Cyt1A in these recombinants, which synergizes Cry toxicity and delays resistance to these proteins and Bs Bin, should enable long term use of these recombinants with little if any development of resistance. In the field, these new recombinants should be particularly effective larvicides against most important vectors and nuisance species of the genus Culex, the malaria vectors Anopheles gambiae and An. arabiensis, and species of Aedes and Ochlerotatus sensitive to Bs.

Although most algae are nutritious food for mosquito larvae, some species kill the larvae when ingested in large quantities. Cyanobacteria (blue-green algae) that kill larvae do so by virtue of toxicity. While blue-green algae toxins may offer possibilities for delivery as larvicides, the toxicity of live blue-green algae does not seem consistent enough for live algae to be useful for mosquito control.

Certain species of green algae in the order Chlorococcales kill larvae primarily because they are indigestible. Where these algae are abundant in nature, larvae consume them to the exclusion of other food and then starve. Under the right circumstances, it is possible to introduce indigestible algae into a breeding habitat so they become abundant enough to render it unsuitable for mosquito production. The algae can persist for years, even if the habitat dries periodically. The main limitation of indigestible algae lies in the fact that, under certain conditions, they may not replace all the nutritious algae in the habitat. More research on techniques to ensure complete replacement will be necessary before indigestible algae can go into operational use for mosquito control.

Juvenile turtles have the capacity to eat more than 500 3rd and 4th instar mosquitos per day. Keeping one turtle in each water-storage tank during field trials for a dengue-control project in Honduras eliminated all mosquito production from the tanks. In Louisiana, keeping turtles in residential roadside ditches polluted by septic-tank effluent reduced Culex quinquefasciatus larvae and pupae by more than 99%. Turtles can serve as alternate hosts for Salmonella when kept in small pet containers, but the available evidence indicates that turtles create no Salmonella hazard in water-storage tanks or other mosquito-breeding habitats. Although turtles would probably not be practical for mosquito control in roadside ditches, they could be effective in storm-water catch basins or holding ponds.

A brief overview is presented of the discovery and development of s-methoprene and some other juvenile hormone mimics. The identification of the natural juvenile hormones is described along with an outline of the part they play in the hormonal control of insect development. The properties and commercial applications of s-methoprene are presented with emphasis on its use in mosquito control and its minimal impact on the environment.

Technologies which utilize semiochemicals, traps/targets and mass trapping are relatively new for management of adult mosquito populations. To date most of the emphasis has been on developing barriers of attractant-baited and insecticide-impregnated targets. The most successful continuous use of this type of technology has been at Stevens' Landing, Collier County, FL. Recently, commercially available traps have been evaluated for their ability to reduce nuisance populations of mosquitoes. Use of Mosquito Magnet™ Pro traps along a nature trail on an isolated island (Atsena Otie) in the Gulf of Mexico resulted in a significant reduction in annoyance caused by the black salt marsh mosquito Ochlerotatus taeniorhynchus (Wiedemann). This chapter presents a brief discussion of semiochemicals (behavior modifying) and mosquito traps, strategies for using them for insect control, and an overview of the recent advancements in research activities conducted to evaluate using combinations of semiochemicals and mass trapping techniques for adult mosquito management.

Models can be useful at many different levels when considering complex issues such as biological control of mosquitoes. At an early stage, exploratory models are valuable in exploring the characteristics of an ideal biological control agent and for guidance in data collection. When more data are available, models can be used to explore alternative control strategies and the likelihood of success. There are few modeling studies that explicitly consider biological control in mosquitoes; however, there have been many theoretical studies of biological control in other insect systems and of mosquitoes and mosquito-borne diseases in general. Examples are used here to illustrate important aspects of designing, using and interpreting models. The stability properties of a model are valuable in assessing the potential of a biological control agent, but may not be relevant to a mosquito population with frequent environmental perturbations. The time scale and goal of proposed control strategies are important considerations when analyzing a model. The underlying biology of the mosquito host and the biological control agent must be carefully considered when deciding what to include in a model. Factors such as density dependent population growth in the host, the searching efficiency and aggregation of a natural enemy, and the resource base of both have been shown to influence the stability and dynamics of the interaction. Including existing mosquito control practices into a model is useful if biological control is proposed for locations with current insecticidal control. The development of Integrated Pest Management (IPM) strategies can be enhanced using modeling techniques, as a wide variety of options can be simulated and examined. Models can also be valuable in comparing alternate routes of disease transmission and to investigate the level of control needed to reduce transmission.

This chapter reviews aspects of population dynamics that may be conceptually important for biological control of mosquitoes. Density dependent population regulation among immature stages has important implications for biological control of mosquito populations, primarily because it can lead to compensatory or overcompensatory mortality due to additions of a biological control agent. This can result in control efforts leading to no change in the target population, or actual increases in the target population, respectively. Density dependent effects, and compensatory or overcompensatory mortality, appear to be most common in mosquitoes from container or highly ephemeral habitats. In permanent ground water habitats generalist predators appear to limit mosquito populations and so render mortality additive. Thus, biological control in permanent ground water habitats seems to have the highest likelihood of producing a satisfactory result. A central premise of classical biological control is that pest populations are reduced by enemies to stable equilibrium levels that are both below the pre-control equilibrium level, and well below the level producing detrimental effects. This premise results in predictions that successful biological control is likely to involve specialist enemies (usually parasitoids), with short generation times relative to the victim, high rates of successful search, rapid rates of increase, and needing only a few victims to complete their life cycle. These predictions largely fail for mosquito systems, in which successful biological control seems to be associated with generalist enemies that can kill a large portion of the target population, often causing local extinction, and can persist in the absence of the target organism. Biological control of mosquitoes appears to be inherently unstable, thus contrasting sharply with classical biological control. This review suggests a need for better data on density dependent regulation of mosquito populations.