title: Altitudinal population structure and microevolution of the malaria vector Anopheles cruzii (Diptera: Culicidae).
abstract: BACKGROUND
  In 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.
  
  
  METHODS
  Larvae 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.
  
  
  RESULTS
  Wing 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).
  
  
  CONCLUSIONS
  Wing 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.
authors.affiliation: Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, CEP 05509-300, Brazil. camilalorenz@usp.br.|Biologia da Relação Patógeno-Hospedeiro, Instituto de Ciências Biomédicas, University of Sao Paulo, Av. Prof. Lineu Prestes, 2415, São Paulo, CEP 05508-000, Brazil. camilalorenz@usp.br.
authors.forename: Camila
authors.initials: C
authors.lastname: Lorenz
authors.affiliation: Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, 1500, São Paulo, CEP 05509-300, Brazil. marques.tatiani@hotmail.com.
authors.forename: Tatiani Cristina
authors.initials: TC
authors.lastname: Marques
authors.affiliation: Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, 1500, São Paulo, CEP 05509-300, Brazil. masallum@usp.br.
authors.forename: Maria Anice Mureb
authors.initials: MA
authors.lastname: Sallum
authors.affiliation: Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, CEP 05509-300, Brazil. lincoln.suesdek@butantan.gov.br.|Instituto de Medicina Tropical, Avenida Dr. Enéas Carvalho de Aguiar 470, São Paulo, CEP 05403-000, Brazil. lincoln.suesdek@butantan.gov.br.
authors.forename: Lincoln
authors.initials: L
authors.lastname: Suesdek
mesh_terms: D000531:Altitude; D000818:Animals; D000852:Anopheles / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D005075:Biological Evolution; D001938:Brazil; D003576:Electron Transport Complex IV / Q000235:genetics; D005260:Female; D019476:Insect Proteins / Q000235:genetics; D007303:Insect Vectors / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D008297:Male; D011156:Population Density; D011157:Population Dynamics; D014921:Wings, Animal / Q000254:growth & development / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D003576:Electron Transport Complex IV