Culicidae colonization in laboratory is paramount to conduct studies aiming at a better understanding of mosquitoes' capacity to transmit pathogens that cause deadly diseases. Colonization requires female blood feeding, a necessary step for maturation of female's oocytes. Direct blood feeding on anesthetized mammals implies in a number of disadvantages when compared to artificial blood feeding. Consequently, laboratories worldwide have been trying to -feed female mosquitoes artificially in order to replace direct feeding. In this study, we compared the effects of direct blood feeding and artificial blood feeding on important life traits of three Culicidae species. Artificial feeding was performed using citrated or defibrinated sheep blood and citrated or defibrinated rabbit blood. Direct feeding was performed using anesthetized guinea pigs as the blood source and the experiment control. Results indicated that artificial feeding using sheep blood was not good enough to justify its use in the maintenance of laboratory colonies of Culicidae. However, artificial feeding using rabbit blood maintained a recovery rate always very close to the control, especially when blood was citrated. We concluded that artificial feeding using citrated rabbit blood can substitute direct feeding on mammals reducing the use of animals, eliminating the need to maintain a bioterium in the laboratory and reducing costs in scientific researches involving Culicidae vectors.

Aedes aegypti is the vector responsible for transmitting pathogens that cause various infectious diseases, such as dengue, Zika, yellow fever, and chikungunya, worrying health authorities in the tropics. Due to resistance of mosquitoes to synthetic insecticides, the search for more effective insecticidal agents becomes crucial. The aim of this study was to verify the larvicidal, adulticidal, and anticholinesterase activities of the essential oils of the Illicium verum (EOIV), Pimenta dioica (EOPD), and Myristica fragrans (EOMF) against Ae. aegypti. The essential oils (EOs) were obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The larvicidal and adulticidal activities of EOs were evaluated against third instar larvae and Ae. aegypti adult females, respectively, using the procedures of the World Health Organization (WHO) and the anticholinesterase activity of the EOs by the modified Ellman method. The following major components were identified: (E)-anethole (90.1%) for EOIV, methyl eugenol (55.0%) for EOPD, and sabinene (52.1%) for EOMF. All EOs exhibited larvicidal and adulticidal activity against Ae. aegypti. The highest larval mortality was observed in EOMF with LC = 28.2 μg mL. Adult mortality was observed after 1 (knockdown) and 24 h exposure, with the highest potential established by the EOIV, KC = 7.3 μg mg female and LC = 10.3 μg mg female. EOIV (IC = 4800 μg mL), EOMF (IC = 4510 μg mL), and EOPD (IC = 1320 μg mL) inhibited AChE. EOMF (4130 μg mL) and EOPD (IC = 3340 μg mL) inhibited BChE whereas EOIV showed no inhibition. The EOs were toxic to larvae and adults of Ae. aegypti, as well as being less toxic to humans than the currently used insecticides, opening the possibility of elaboration of a natural, safe, and ecological bioinsecticide for vector control.

Background Chemical mosquito control using malathion has been applied in Brazil since 1985. To obtain chemical control effectiveness, vector susceptibility insecticide monitoring is required. This study aimed to describe bioassay standardizations and determine the susceptibility profile of Ae. aegypti populations to malathion and pyriproxyfen, used on a national scale in Brazil between 2017 and 2018, and discuss the observed impacts in arbovirus control. Methods The diagnostic-doses (DD) of pyriproxyfen and malathion were determined as the double of adult emergence inhibition (EI) and lethal doses for 99% of the Rockefeller reference strain, respectively. To monitor natural populations, sampling was performed in 132 Brazilian cities, using egg traps. Colonies were raised in the laboratory for one or two generations (F1 or F2) and submitted to susceptibility tests, where larvae were exposed to the pyriproxyfen DD (0.03 µg/l) and adults, to the malathion DD determined in the present study (20 µg), in addition to the one established by the World Health Organization (WHO) DD (50 µg) in a bottle assay. Dose-response (DR) bioassays with pyriproxyfen were performed on populations that did not achieve 98% EI in the DD assays. Results Susceptibility alterations to pyriproxyfen were recorded in six (4.5%) Ae. aegypti populations from the states of Bahia and Ceará, with Resistance Ratios (RR95) ranging from 1.51 to 3.58. Concerning malathion, 73 (55.3%) populations distributed throughout the country were resistant when exposed to the local DD 20 µg/bottle. On the other hand, no population was resistant, and only 10 (7.6%) populations in eight states were considered as exhibiting decreased susceptibility (mortality ratios between 90 and 98%) when exposed to the WHO DD (50 µg/bottle). Conclusions The feasibility of conducting an insecticide resistance monitoring action on a nation-wide scale was confirmed herein, employing standardized and strongly coordinated sampling methods and laboratory bioassays. Brazilian Ae. aegypti populations exhibiting decreased susceptibility to pyriproxyfen were identified. The local DD for malathion was more sensitive than the WHO DD for early decreased susceptibility detection.

The mosquito Aedes aegypti is the primary vector of different arboviruses and represents a major public health problem. Several Brazilian populations of Ae. aegypti have developed resistance to temephos, the most used organophosphate larvicide. New tools which are less harmful to the environment and safer for humans are becoming increasingly important to control this insect vector. Spinosad, an aerobic fermentation product of a soil actinobacteria, has a favorable environmental profile. It presents selective insecticide properties, a mechanism of action that differs from those of many synthetic chemical insecticides. The toxicity of spinosad and temephos to Aedes aegypti populations from Brazil, which were previously exposed to temephos, were investigated in this study. Larval susceptibility (LC50) to temephos varied from 3μg/L for Rockefeller up to 260 μg/L for Santana do Ipanema field derived population. Larval susceptibility (LC50) to spinosad varied from 23μg/L for Rockefeller up to 93μg/L for Marilia field derived population. In addition, a semi-field trial was performed to evaluate spinosad (NatularTM DT) initial efficacy and persistence toward four field-derived lineages and the Rockefeller lineage, used as an internal control. Spinosad was tested at 0.5mg active ingredient/L in 200L capacity water tanks. Mortality was recorded each 24 hours after exposition and tanks were further recolonized once per week with mortality being recorded daily for eight weeks. Spinosad provided a level equal or superior to 80% mortality during a seven to eight week evaluation period. The assessed populations did not present cross-resistance between spinosad and temephos in laboratory conditions. It demonstrates that spinosad may be a promising larvicide for the control of Ae. aegypti, especially for populations in which resistance to temephos has been detected.