*4.4. Study Limitations*

The study has several limitations which should be addressed in subsequent work. Firstly, experiments were conducted in a single testing facility. A comparison of the alternative method in multiple laboratories is desirable to ensure the reproducibility of the methods with other mosquito strains. The low feeding success with the membrane technique needs to be overcome, as clearly feeding success impacts mosquito mortality. Ideally, the membrane bioassay will be improved to consistently measure 50% mosquito feeding success at multiple testing facilities. Additionally, two different ITN products from the same manufacturer were used. It could be argued that the evaluation of dual AI nets of pyrethroids with PBO and PPF would also be as relevant, although these are best measured using WHO cone tests as they do not require "free-flying" bioassays for evaluation. Similar experiments conducted by other facilities are recommended to generate further evidence of the range of values and precision of the estimates of mortality and blood feeding inhibition using the 50-rabbit and 50-membrane technique.
