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Assessing the molecular divergence between Anopheles (Kerteszia) cruzii populations from Brazil using the timeless gene: further evidence of a species complex.

ABSTRACT: Anopheles (Kerteszia) cruzii was the most important vector of human malaria in southern Brazil between 1930-1960. Nowadays it is still considered an important Plasmodium spp. vector in southern and south-eastern Brazil, incriminated for oligosymptomatic malaria. Previous studies based on the analysis of X chromosome banding patterns and inversion frequencies in An. cruzii populations from these areas have suggested the occurrence of three sibling species. In contrast, two genetically distinct groups among An. cruzii populations from south/south-east and north-east Brazil have been revealed by isoenzyme analysis. Therefore, An. cruzii remains unclear.In this study, a partial sequence of the timeless gene (approximately 400 bp), a locus involved in the control of circadian rhythms, was used as a molecular marker to assess the genetic differentiation between An. cruzii populations from six geographically distinct areas of Brazil.The timeless gene revealed that An. cruzii from Itaparica Island, Bahia State (north-east Brazil), constitutes a highly differentiated group compared with the other five populations from south and south-east Brazil. In addition, significant genetic differences were also observed among some of the latter populations.Analysis of the genetic differentiation in the timeless gene among An. cruzii populations from different areas of Brazil indicated that this malaria vector is a complex of at least two cryptic species. The data also suggest that further work might support the occurrence of other siblings within this complex in Brazil.

SUBMITTER: Rona LD

PROVIDER: S-EPMC2673228 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature

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Assessing the molecular divergence between Anopheles (Kerteszia) cruzii populations from Brazil using the timeless gene: further evidence of a species complex.

Rona Luísa D P LD Carvalho-Pinto Carlos J CJ Gentile Carla C Grisard Edmundo C EC Peixoto Alexandre A AA

Malaria journal 20090409

<h4>Background</h4>Anopheles (Kerteszia) cruzii was the most important vector of human malaria in southern Brazil between 1930-1960. Nowadays it is still considered an important Plasmodium spp. vector in southern and south-eastern Brazil, incriminated for oligosymptomatic malaria. Previous studies based on the analysis of X chromosome banding patterns and inversion frequencies in An. cruzii populations from these areas have suggested the occurrence of three sibling species. In contrast, two gene ...[more]

PMID: 19358734

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Project description:BackgroundThe mosquito Anopheles (Kerteszia) cruzii is the main vector of human and simian malaria in the Atlantic Forest. This species is usually abundant in the forests where it occurs, preferring to live and feed on canopies, behaviour known as acrodendrophily. However, in several studies and locations this species has been observed in high density near the ground in the forest. In this study, it was hypothesized that factors associated with anthropogenic landscape changes may be responsible for the variation in abundance and acrodendrophily observed in An. cruzii.MethodsThe study was conducted in a conservation unit in the city of São Paulo, Brazil. Monthly entomological collections were performed from March 2015 to April 2017, and the resulting data were used with data from another study conducted in the same area between May 2009 and June 2010. Mosquitoes were collected from five sites using CDC and Shannon traps. Landscape composition and configuration metrics were measured, and generalized linear mixed-effect models were used to investigate the relationship between these metrics and variations in the abundance and acrodendrophily of An. cruzii.ResultsThe model that showed the best fit for the relationship between landscape metrics and An. cruzii abundance suggests that an increase in the proportion of forest cover leads to an increase in the abundance of this mosquito, while the model that best explained variations in An. cruzii acrodendrophily suggests that an increase in total forest-edge length leads to greater activity by this species at ground level.ConclusionWhile the data indicate that changes in landscape due to human activities lead to a reduction in An. cruzii abundance, such changes may increase the contact rate between this species and humans living on the edges of forest fragments where An. cruzii is found. Future studies should, therefore, seek to elucidate the effect of these landscape changes on the dynamics of Plasmodium transmission in the Atlantic Forest, which according to some studies includes the participation of simian hosts.

Project description:Cryptic species complexes are common among anophelines. Previous phylogenetic analysis based on the complete mtDNA COI gene sequences detected paraphyly in the Neotropical malaria vector Anopheles marajoara. The "Folmer region" detects a single taxon using a 3% divergence threshold.To test the paraphyletic hypothesis and examine the utility of the Folmer region, genealogical trees based on a concatenated (white + 3' COI sequences) dataset and pairwise differentiation of COI fragments were examined. The population structure and demographic history were based on partial COI sequences for 294 individuals from 14 localities in Amazonian Brazil. 109 individuals from 12 localities were sequenced for the nDNA white gene, and 57 individuals from 11 localities were sequenced for the ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2).Distinct A. marajoara lineages were detected by combined genealogical analysis and were also supported among COI haplotypes using a median joining network and AMOVA, with time since divergence during the Pleistocene (<100,000 ya). COI sequences at the 3' end were more variable, demonstrating significant pairwise differentiation (3.82%) compared to the more moderate 2.92% detected by the Folmer region. Lineage 1 was present in all localities, whereas lineage 2 was restricted mainly to the west. Mismatch distributions for both lineages were bimodal, likely due to multiple colonization events and spatial expansion (~798-81,045 ya). There appears to be gene flow within, not between lineages, and a partial barrier was detected near Rio Jari in Amapá state, separating western and eastern populations. In contrast, both nDNA data sets (white gene sequences with or without the retention of the 4th intron, and ITS2 sequences and length) detected a single A. marajoara lineage.Strong support for combined data with significant differentiation detected in the COI and absent in the nDNA suggest that the divergence is recent, and detectable only by the faster evolving mtDNA. A within subgenus threshold of >2% may be more appropriate among sister taxa in cryptic anopheline complexes than the standard 3%. Differences in demographic history and climatic changes may have contributed to mtDNA lineage divergence in A. marajoara.

Project description:BackgroundIn Brazil, the autochthonous transmission of extra-Amazonian malaria occurs mainly in areas of the southeastern coastal Atlantic Forest, where Anopheles cruzii is the primary vector. In these locations, the population density of the mosquito varies with altitude (5-263 m above sea level), prompting us to hypothesise that gene flow is also unevenly distributed. Describing the micro-geographical and temporal biological variability of this species may be a key to understanding the dispersion of malaria in the region. We explored the homogeneity of the An. cruzii population across its altitudinal range of distribution using wing shape and mtDNA gene analysis. We also assessed the stability of wing geometry over time.MethodsLarvae were sampled from lowland (5-20 m) and hilltop (81-263 m) areas in a primary Atlantic Forest region, in the municipality of Cananéia (State of São Paulo, Brazil). The right wings of males and females were analysed by standard geometric morphometrics. Eighteen landmarks were digitised for each individual and a discriminant analysis was used to compare samples from the hilltop and lowland. A 400-bp DNA fragment of the mitochondrial cytochrome oxidase gene subunit I (CO-I) was PCR-amplified and sequenced.ResultsWing shapes were distinct between lowland and hilltop population samples. Results of cross-validated tests based on Mahalanobis distances showed that the individuals from both micro-environments were correctly reclassified in a range of 54-96%. The wings of hilltop individuals were larger. The CO-I gene was highly polymorphic (haplotypic diversity = 0.98) and altitudinally structured (Фst = 0.085 and Jaccard = 0.033). We found 60 different haplotypes but only two were shared by the lowland and hilltop populations. Wing shape changed over the brief study period (2009-2013).ConclusionsWing geometry and CO-I gene analysis indicated that An. cruzii is vertically structured. Wing shape varied rapidly, but altitude structure was maintained. Future investigations should identify the biotic/abiotic causes of these patterns and their implications in the local epidemiology of malaria.

Dataset Information

Assessing the molecular divergence between Anopheles (Kerteszia) cruzii populations from Brazil using the timeless gene: further evidence of a species complex.

Publications

Assessing the molecular divergence between Anopheles (Kerteszia) cruzii populations from Brazil using the timeless gene: further evidence of a species complex.

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