Identification of a fibrinogen-related protein (FBN9) gene in neotropical anopheline mosquitoes.

BACKGROUND: Malaria has a devastating impact on worldwide public health in many tropical areas. Studies on vector immunity are important for the overall understanding of the parasite-vector interaction and for the design of novel strategies to control malaria. A member of the fibrinogen-related protein family, fbn9, has been well studied in Anopheles gambiae and has been shown to be an important component of the mosquito immune system. However, little is known about this gene in neotropical anopheline species. METHODS: This article describes the identification and characterization of the fbn9 gene partial sequences from four species of neotropical anopheline primary and secondary vectors: Anopheles darlingi, Anopheles nuneztovari, Anopheles aquasalis, and Anopheles albitarsis (namely Anopheles marajoara). Degenerate primers were designed based on comparative analysis of publicly available Aedes aegypti and An. gambiae gene sequences and used to clone putative homologs in the neotropical species. Sequence comparisons and Bayesian phylogenetic analyses were then performed to better understand the molecular diversity of this gene in evolutionary distant anopheline species, belonging to different subgenera. RESULTS: Comparisons of the fbn9 gene sequences of the neotropical anophelines and their homologs in the An. gambiae complex (Gambiae complex) showed high conservation at the nucleotide and amino acid levels, although some sites show significant differentiation (non-synonymous substitutions). Furthermore, phylogenetic analysis of fbn9 nucleotide sequences showed that neotropical anophelines and African mosquitoes form two well-supported clades, mirroring their separation into two different subgenera. CONCLUSIONS: The present work adds new insights into the conserved role of fbn9 in insect immunity in a broader range of anopheline species and reinforces the possibility of manipulating mosquito immunity to design novel pathogen control strategies.

Malaria vectors in areas of Plasmodium falciparum epidemic transmission in the Amazon region, Brazil.

The main vectors of malaria in Brazil are Anopheles darlingi, An. aquasalis, and some species of the An. albitarsis complex, whereas others have questionable importance with regard to the disease transmission. To identify these vectors in the State of Pará, Brazil, in a high-prevalence P. falciparum area, 565 anophelines were captured and identified while the seasonal variation and daily biting activity were determined. Of the seven anopheline species (An. strodei, An. albitarsis s.l., An. rondoni, An. darlingi, An. triannulatus, An. oswaldoi, and An. nuneztovari), the plasmodia circumsporozoite protein (CSP) was detected in three of them, with a total infection rate of 6.2%. An. darlingi was the most prevalent species (22.4%), followed by An. albitarsis (5.2%) and An. rondoni (3.6%). An. rondoni was found to be infected for the first time, which was also confirmed through PCR. This result possibly represents a new malaria vector based on its highest frequency, biting and seasonal activities in the peak of malaria transmission.