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Due to the absence of a perfect method for mosquito sex separation, the combination of the sterile insect technique and the incompatible insect technique is now being considered as a potentially effective method to control  Aedes albopictus . In this present study first we examine the minimum pupal irradiation dose required to induce complete sterility in  Wolbachia  triple-infected (HC), double-infected (GUA) and uninfected (GT) female  Ae. albopictus . The HC line is a candidate for  Ae. albopictus  population suppression programmes, but due to the risk of population replacement which characterizes this triple infected line, the individuals to be released need to be additionally irradiated. After determining the minimum irradiation dose required for complete female sterility, we test whether sterilization is sufficient to prevent invasion of the triple infection from the HC females into double-infected (GUA) populations. Our results indicate that irradiated  Ae. albopictus  HC, GUA and GT strain females have decreased fecundity and egg hatch rate when irradiated, inversely proportional to the dose, and the complete sterilization of females can be acquired by pupal irradiation with doses above 28 Gy. PCR-based analysis of F 1  and F 2  progeny indicate that the irradiated HC females, cannot spread the new  Wolbachia w Pip strain into a small cage GUA population, released at a 1:5 ratio. Considering the above results, we conclude that irradiation can be used to reduce the risk of population replacement caused by an unintentional release of  Wolbachia  triple-infected  Ae. albopictus  HC strain females during male release for population suppression.

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Combining the Sterile Insect Technique with Wolbachia-Based Approaches: II- A Safer Approach to Aedes albopictus Population Suppression Programmes, Designed to Minimize the Consequences of Inadvertent Female Release