**2.3 Maintenance of immature and adult of** *A. aegypti* **from Londrina, Paraná, under laboratory conditions**

The reeds containing eggs were immersed in plastic trays (45 × 30 × 7.5 cm) containing distilled water to stimulate the larvae to hatch. The immatures obtained were kept until adults by means of food containing a mixture of cat food (Whiskas®) and rodents (Teklab global®) in a 1:1 ratio, ground into fine particles (1 mm). All trays were covered with a nylon fabric to prevent the escape of mosquitoes. After emergence, the males and females were collected with a Castro catcher and the species identification was performed using external morphological characters, mainly from the chest, with the aid of a stereoscopic microscope ZEISS Stemi 2000 50× and identification keys proposed by Forattini [14], Harbach [24], and WRBU [25].

After the identification stage, the adults were placed to copulate in cardboard cages (17 × 20 cm), containing two plastic cups lined with strips of filter paper and filled with 70 mL of distilled water, which were used as a substrate for oviposition. As a source of carbohydrates, an Erlenmeyer was introduced containing a roll of gauze with pieces of cotton in the center, soaked with 12% sugar water. The blood meal was carried out using an anesthetized hamster for 30min, according to the procedure approved by the Ethics Committee on the Use of Animals at UEL ("Breeding of mosquitoes in laboratory conditions").

### **2.4 Biological aspects of** *A. aegypti* **from Londrina, Paraná, in incubator chambers (BOD) with different temperatures**

At UEL, the experiment was carried out in four incubator chambers (BOD) that had different temperatures (0, 5, 25 and 45°C) with ±2°C, as well as at ambient temperature, where the eggs remained in dry incubation in 500 ml capacity pots containing only moistened cotton over a period of 10 days, with a 12/12 h photoperiod.

Subsequently, 150 ml of distilled water, 25 eggs of *A. aegypti*, F1 field generation and 0.055 g of larval food were introduced into each of the five pots, which were placed in BOD with temperature of 25 ± 2°C. The monitoring of the hatching rate of eggs, the rate of immature deaths and the number of adults were carried out daily.
