After feeding the female mosquitoes with blood for 3 days, the
oviposition containers that were in the breeding cages were
removed. To synchronize hatching, the filter paper was removed and left to dry for 3 days at room temperature for the
embryos to mature; following this period, hatching was stimulated to obtain the larvae for the experiments. When they
reached the larval stage, between the third and fourth instar,
they were counted, separated, and transferred with Pasteur
pipettes to plastic cups with 100 mL of the same water without
chlorine, where a diagnostic assay (DA) was conducted,
which determined the concentrations that cause mortality
between 2 and 98 % in the larvae of A. aegypti. For the
experiment DA, a total of 120 larvae distributed in three
concentrations (1,000, 300, and 30 ppm) were used with four
replicas each, along with control treatment without the evaluated oil and with DMSO at 0.5 %. Mortality was assessed by
recording the larvae that were unable to reach the surface of the water when the beaker of the experiment was tapped; they
were considered dead (WHO 1981, 1992). Subsequently, five
concentrations were established for each oil. In each, four
replicas were conducted per concentration in addition to the
control treatment, which was the same as that employed to
determine those of the DA. Each completed experiment was
repeated three times on different days. Larval mortality readngs were taken at two times, one at 24 h and the other at 48 h.
The results of mortality and survival of the bioassay were
subjected to probit analysis