**2.5 Stabilization of a sample of the population of** *A. aegypti* **from Manaus, Amazonas, in environmental rooms with temperature and CO2 conditions according to the provisions of the IPCC**

The same process of creating and maintaining the immature and adults of *A. aegypti* from Londrina, Paraná, mentioned in items 2.2 and 2.3, was carried out with a sample of the population of *A. aegypti* from Manaus, Amazonas, collected in the same way on *Campus* I of INPA. However, the blood meal carried out followed the procedure approved by the Ethics Committee on the Use of Animals at INPA (CEUA/INPA: 04/2013-"Breeding of mosquito vectors, under laboratory conditions").

After the stabilization of the three insectaries in the LEEM, the F28 generation was obtained in the different environmental rooms that started the experiment for post-embryonic development, using three different environments. Rooms 1, 2 and 4 suffer from interference from different temperatures and levels of carbon dioxide, and rooms 2 and 4 refer to 50 and 100 years in front of room 1, which presents ambient temperature.

These rooms are equipped with technological devices that guarantee, respectively, the maintenance of ambient temperature, 2.5°C (400 ppm CO2) and 4.5°C (850 ppm CO2) above ambient temperature. The natural conditions of the respective environmental rooms are collected in real time by sensors isolated in the forest and the humidity of all remains constant in relation to the environmental conditions.
