In the Americas, areas with a high risk of malaria transmission are mainly located in the Amazon Forest, which extends across nine countries. One keystone step to understanding the Plasmodium life cycle in Anopheles species from the Amazon Region is to obtain experimentally infected mosquito vectors. Several attempts to colonise Ano- pheles species have been conducted, but with only short-lived success or no success at all. In this review, we review the literature on malaria transmission from the perspective of its Amazon vectors. Currently, it is possible to develop experimental Plasmodium vivax infection of the colonised and field-captured vectors in laboratories located close to Amazonian endemic areas. We are also reviewing studies related to the immune response to P. vivax infection of Anopheles aquasalis, a coastal mosquito species. Finally, we discuss the importance of the modulation of Plasmodium infection by the vector microbiota and also consider the anopheline genomes. The establishment of experimental mosquito infections with Plasmodium falciparum, Plasmodium yoelii and Plasmodium berghei parasites that could provide interesting models for studying malaria in the Amazonian scenario is important. Understanding the molecular mechanisms involved in the development of the parasites in New World vectors is crucial in order to better determine the interaction process and vectorial competence.

Anopheles darlingi is the major malaria vector in countries located in the Amazon region. Anopheles aquasalis and Anopheles albitarsis s.l. are also proven vectors in this region. Anopheles nuneztovari s.l. and Anopheles triannulatus s.l. were found infected with Plasmodium vivax; however, their status as vectors is not yet well defined. Knowledge of susceptibility of Amazon anopheline populations to Plasmodium infection is necessary to better understand their vector capacity. Laboratory colonization of An. darlingi, the main Amazon vector, has proven to be difficult and presently An. aquasalis is the only available autonomous colony.

The eggs of three Amazonian species of the genus Mansonia (Diptera: Culicidae) were analyzed using morphological and morphometric characters. Eggs of Mansonia humeralis Dyar & Knab, 1916 were morphologically different from those of Mansonia titillans (Walker, 1848) and Mansonia amazonensis (Theobald, 1901), which were more similar to each other according to linear and geometric morphometry. A principal component analysis generated from elliptic Fourier contour data (PC1—92.6% and PC2—2.61%) indicated that Ma. amazonensis and Ma. titillans are more similar to each other than either is to Ma. humeralis. Discriminant multivariate analysis was highly accurate with only four classification errors and a 90% accuracy rate. The results indicate that the three Amazonian species can be precisely distinguished in the egg stage and that geometric morphometry based on elliptic Fourier contours is a promising method for distinguishing eggs of species of Mansonia. An identification key based on egg morphology is provided to distinguish the four Neotropical species.

Aedes aegypti is currently controlled with synthetic larvicides; however, mosquitoes have become highly resistant to these larvicides and difficult to eradicate. Studies have shown that insecticides derived from fungal extracts have various mechanisms of action that reduce the risk of resistance in these mosquitoes. One possible mechanism is uncontrolled production of reactive oxygen species (ROS) in the larvae, which can cause changes at the cellular level. Thus, the crude extract of Xylaria sp. was evaluated to investigate the oxidative effect of this extract in A. aegypti larvae by quantifying the oxidative damage to proteins and lipids. The larvicidal potential of the crude extract of Xylaria sp. Was evaluated, and the extract was subsequently tested in human lung fibroblasts for cytotoxicity and ROS production. ROS level was quantified in the larvae that were killed following exposure to the extract in the larvicide test. The crude extract of Xylaria sp. Caused cytotoxicity and induced ROS production in human lung fibroblasts and A. aegypti larvae, respectively. In the larvicide trial, the extract showed an LC₅₀ of 264.456 ppm and an LC90 of 364.307 ppm, and was thus considered active. The extract showed greater oxidative damage to lipids and proteins, with LC₉₀ values of 24.7 µmol MDA/L and 14.6278 ×10^-3 nmol carbonyl/ mg protein, respectively. Crude extracts of Xylaria sp. induced oxidative stress that may have caused the mortality of A. aegypti larvae.

Aedes aegypti is the primary vector of viruses, such as Zika, chikungunya, yellow fever, and dengue. In this context, a biomonitored chemical study was conducted to evaluate the activity of the crude extract of the endophytic fungus Phomopsis sp. against the larvae of Aedes aegypti. Crude extract, fractions, and isolated substances were evaluated in in-vitro assays against third-stage larvae of Aedes aegypti. We isolated 3-nitropropionic acid with an LC₅₀ of 15.172 ppm and LC₉₀ of 18.178 ppm after 24 hours of larval exposure. The results indicated that 3-nitropropionic acid exerted larvicidal activity.

Aedes aegypti and Anopheles darlingi represent health challenges due to synthetic insecticide resistance. Hence, the essential oil from Piper humillimum is an alternative for vector control. In this study, the essential oil (3.5 ± 0.4% yield) alongside germacrene D (61.51%) and δ-cadinene (17.46%) showed larvicidal activity (LC₅₀ of 34.75 to 46.04 µg/mL), accompanied by an increase of hydrogen peroxide (H₂O₂) production (36.67 ± 1.52 to 81.33 ± 1.52 µmol H₂O₂ min⁻¹ mg⁻¹ protein), causing lipid (43.3 ± 6.02 to 81.67 ± 3.05 nmol malondialdehyde mg⁻¹ protein) and protein damages (61.67 ± 6.80 to 83.00 ± 2.64 nmol carbonyls mg⁻¹ protein). Further triggering an increase in superoxide dismutase (83.31 ± 6.80 to 95.00 ± 3.60 U mg⁻¹ protein) and catalase (74.31 ± 7.02 to 82.09 ± 1.00 µmol H₂O₂ min⁻¹ mg⁻¹ protein) activities. In addition, mixed-function oxidases (61.17 ± 11.37 to 73.52 ± 6.42 nmol cyt c min⁻¹ mg⁻¹ protein), α- and β-esterase (38.41 ± 4.04 to 61.31 ± 9.29 µmol min⁻¹ mg⁻¹ protein) levels increased. Conversely, glutathione S-transferase (GST) (11.01 ± 2.00 to 9.67 ± 3.05 µmol min⁻¹ mg⁻¹ protein) and acetylcholinesterase (AChE) (14.33 ± 3.78 to 17.00 ± 1.00 μmol min⁻¹ mg⁻¹ protein) were inhibited, corroborated by molecular docking, with germacrene D and δ-cadinene showing binding energies of −7.9 and −7.9 kcal/mol, 1.63 and 1.94 Ki for AChE, while for GST were −6.4 and 6.6 kcal/mol, and 20.5 and 15.50 Ki, respectively. These results demonstrate that the essential oil from P. humillimum is a promising multi-target alternative for the control of the investigated vectors.