**3. Innovations to biological and genetic manipulation of mosquito vectors**

The strategies for genetic and biological control/manipulation with *Wolbachia* of mosquito vectors (GMM/BCMW) propose an attack on the mechanisms directly responsible for the proliferation of mosquito populations. Allowing the mosquitoes' reproductive dynamics be the tool for spreading the intervention means that we will allow the modified populations to disperse naturally (through repeated releases) so that little by little the mosquitoes go about occupying the territory of wild populations to the point of reaching our objective by replacing them in their function as vectors or by suppressing them as a species.

The mechanism of dispersion and coverage that is proposed is the male mosquito vector itself; these male mosquitoes will find their female counterparts and transmit the control measure before these females lay their eggs, undiscriminating as to preferred breeding site and location. The progeny (eggs, larvae and adults) will incorporate the intervention naturally and will maintain it in the population that emerges from their lineage (desirable). In essence, the dispersal and upkeep of the intervention will be a product of biological mechanisms rather than human intervention.

Interventions consisting of biological manipulation and genetic control of vectors, furthermore, share many characteristics that again distinguish them from the traditional methods. Among these are as follows: (1) dependence on vertical (maternal) transmission of heritable elements (resistance genes and *Wolbachia*), (2) specificity in regard to affected species, (3) environmental friendliness, (4) harnessing of natural reproductive instincts, (5) noninvasiveness of domestic spaces and (6) large-scale application (indispensable). A common challenge of these innovations and of traditional measures of control is to achieve the coverage necessary to be effective and sustainable.

In general, these innovations to vector manipulation are based on two strategies that can be organized according to the results obtained (population elimination vs. replacement) or to the implantation dynamics (self-sustainable or self-limiting).

*Population elimination/suppression*: aimed to affect the demographics of the vector population with the goal of eliminating it from the area or reducing it to a low level that will not maintain transmission.

*Population substitution/replacement*: This strategy seeks to replace wild populations with modified populations that are resistant to the viral infection. One of the most novel mechanisms that produce resistance to infection is transinfection with *Wolbachia*. Other mechanisms are effected through the incorporation of transgenes that—by way of impacting the vectors' survival, physiology (flight, feeding) or susceptibility to the infection—indirectly reduce the mosquito's vectorial competence (interference).

*Self-limiting*: This strategy implicates the abundant and repeated release of mosquitoes in order to maintain the flux of the genetic change in the target population. It is reversible with the discontinuation of releases.

*Self-sustaining*: This strategy proposes repeated releases of modified mosquito populations sufficient to establish themselves as the dominant population (replacement), to the end of their persisting in the population even while there may be unforeseen risks.
