Morphometric Wing Characters as a Tool for Mosquito Identification

Mosquitoes are responsible for the transmission of important infectious diseases, causing millions of deaths every year and endangering approximately 3 billion people around the world. As such, precise identification of mosquito species is crucial for an understanding of epidemiological patterns of disease transmission. Currently, the most common method of mosquito identification relies on morphological taxonomic keys, which do not always distinguish cryptic species. However, wing geometric morphometrics is a promising tool for the identification of vector mosquitoes, sibling and cryptic species included. This study therefore sought to accurately identify mosquito species from the three most epidemiologically important mosquito genera using wing morphometrics. Twelve mosquito species from three epidemiologically important genera (Aedes, Anopheles and Culex) were collected and identified by taxonomic keys. Next, the right wing of each adult female mosquito was removed and photographed, and the coordinates of eighteen digitized landmarks at the intersections of wing veins were collected. The allometric influence was assessed, and canonical variate analysis and thin-plate splines were used for species identification. Cross-validated reclassification tests were performed for each individual, and a Neighbor Joining tree was constructed to illustrate species segregation patterns. The analyses were carried out and the graphs plotted with TpsUtil 1.29, TpsRelw 1.39, MorphoJ 1.02 and Past 2.17c. Canonical variate analysis for Aedes, Anopheles and Culex genera showed three clear clusters in morphospace, correctly distinguishing the three mosquito genera, and pairwise cross-validated reclassification resulted in at least 99% accuracy; subgenera were also identified correctly with a mean accuracy of 96%, and in 88 of the 132 possible comparisons, species were identified with 100% accuracy after the data was subjected to reclassification. Our results showed that Aedes, Culex and Anopheles were correctly distinguished by wing shape. For the lower hierarchical levels (subgenera and species), wing geometric morphometrics was also efficient, resulting in high reclassification scores.

[1]  W. Almirón,et al.  COI barcode versus morphological identification of Culex ( Culex ) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil , 2013, Memorias do Instituto Oswaldo Cruz.

[2]  David E Sanin,et al.  Sm16, a major component of Schistosoma mansoni cercarial excretory/secretory products, prevents macrophage classical activation and delays antigen processing , 2015, Parasites & Vectors.

[3]  I. Kitching,et al.  Phylogeny and classification of the Culicidae (Diptera) , 1998 .

[4]  Francisco Chiaravalloti Neto,et al.  Presença de culicídeos em município de porte médio do Estado de São Paulo e risco de ocorrência de febre do Nilo Ocidental e outras arboviroses , 2011 .

[5]  Rotraut A. G. B. Consoli,et al.  Principais mosquitos de importância sanitária no Brasil , 1994 .

[6]  F. Virginio,et al.  Wing sexual dimorphism of pathogen-vector culicids , 2015, Parasites & Vectors.

[7]  Population genetics of neotropical Culex quinquefasciatus (Diptera: Culicidae) , 2014, Parasites & Vectors.

[8]  Morgan Mangeas,et al.  Climate-Based Models for Understanding and Forecasting Dengue Epidemics , 2012, PLoS neglected tropical diseases.

[9]  Dina M Fonseca,et al.  "Bird biting" mosquitoes and human disease: a review of the role of Culex pipiens complex mosquitoes in epidemiology. , 2011, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[10]  Ted R. Schultz,et al.  Making Mosquito Taxonomy Useful: A Stable Classification of Tribe Aedini that Balances Utility with Current Knowledge of Evolutionary Relationships , 2015, PloS one.

[11]  Dennis E. Slice,et al.  Contributions of Morphometrics to Medical Entomology , 2006 .

[12]  M. T. Marrelli,et al.  Diversity and abundance of mosquitoes (Diptera:Culicidae) in an urban park: larval habitats and temporal variation. , 2015, Acta tropica.

[13]  Dramane Kaba,et al.  The exchangeability of shape , 2010, BMC Research Notes.

[14]  妙子 森保,et al.  世界リンパ系フィラリア症制圧計画(Global Programme to Eliminate Lymphatic Filariasis: GPELF)の政策の枠組み~ゴールの設定と道筋~ , 2018 .

[15]  J. Dujardin,et al.  Geometric morphometrics for the taxonomy of 11 species of Anopheles (Nyssorhynchus) mosquitoes , 2015, Medical and veterinary entomology.

[16]  L. F. Chaves,et al.  Climate Change and Highland Malaria: Fresh Air for a Hot Debate , 2010, The Quarterly Review of Biology.

[17]  W. Almirón,et al.  Discrimination of four Culex (Culex) species from the Neotropics based on geometric morphometrics , 2015, Zoomorphology.

[18]  D. Weetman,et al.  Loss of genetic diversity in Culex quinquefasciatus targeted by a lymphatic filariasis vector control program in Recife, Brazil. , 2011, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[19]  P. Martens,et al.  Climatic-driven seasonality of emerging dengue fever in Hanoi, Vietnam , 2014, BMC Public Health.

[20]  Jun Yang,et al.  Predicting Unprecedented Dengue Outbreak Using Imported Cases and Climatic Factors in Guangzhou, 2014 , 2015, PLoS neglected tropical diseases.

[21]  A. Cornel,et al.  Culex pipiens Sensu Lato in California: A Complex Within a Complex? , 2012, Journal of the American Mosquito Control Association.

[22]  S. Moore,et al.  Evaluating preservation methods for identifying Anopheles gambiae s.s. and Anopheles arabiensis complex mosquitoes species using near infra-red spectroscopy , 2015, Parasites & Vectors.

[23]  A. R. Medeiros-Sousa,et al.  Mosquito Faunal Survey In a Central Park of the City of São Paulo, Brazil , 2015, Journal of the American Mosquito Control Association.

[24]  I. Kitching,et al.  Phylogeny of mosquitoes of tribe Culicini (Diptera: Culicidae) based on morphological diversity , 2012 .

[25]  A. Wilke,et al.  Microsatellite loci cross-species transferability in Aedes fluviatilis (Diptera:Culicidae): a cost-effective approach for population genetics studies , 2015, Parasites & Vectors.

[26]  F. Chiaravalloti Neto,et al.  [The presence of Culicidae species in medium-sized cities in the State of São Paulo, Brazil and the risk of West Nile fever and other arbovirus infection]. , 2011, Revista da Sociedade Brasileira de Medicina Tropical.

[27]  M. T. Marrelli,et al.  Genetic-morphometric variation in Culex quinquefasciatus from Brazil and La Plata, Argentina. , 2010, Memorias do Instituto Oswaldo Cruz.

[28]  M. B. de Paula,et al.  Mosquito Fauna in Municipal Parks of São Paulo City, Brazil: A Preliminary Survey , 2013, Journal of the American Mosquito Control Association.

[29]  L. Suesdek,et al.  Comparison of wing geometry data and genetic data for assessing the population structure of Aedes aegypti. , 2012, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[30]  O. P. Forattini,et al.  Principais mosquitos de importância sanitária no Brasil , 1995 .

[31]  M. Quiñones,et al.  Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from Southern and Northwest Colombia. , 2002, Memorias do Instituto Oswaldo Cruz.

[32]  O Hammer-Muntz,et al.  PAST: paleontological statistics software package for education and data analysis version 2.09 , 2001 .

[33]  A. Clements,et al.  National spatial and temporal patterns of notified dengue cases, Colombia 2007–2010 , 2014, Tropical medicine & international health : TM & IH.

[34]  L. F. Chaves,et al.  Nonlinear impacts of climatic variability on the density‐dependent regulation of an insect vector of disease , 2012 .

[35]  L. Suesdek,et al.  Microevolution of Aedes aegypti , 2015, PloS one.

[36]  L. Suesdek,et al.  Wing diagnostic characters for Culex quinquefasciatus and Culex nigripalpus (Diptera, Culicidae) , 2011 .

[37]  A. Krüger,et al.  The use of Morphometric Wing Characters to Discriminate Female Culex pipiens and Culex torrentium , 2014, Journal of vector ecology : journal of the Society for Vector Ecology.

[38]  J. Dujardin,et al.  Wing shape of dengue vectors from around the world. , 2010, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[39]  Ø. Hammer,et al.  PAST: PALEONTOLOGICAL STATISTICAL SOFTWARE PACKAGE FOR EDUCATION AND DATA ANALYSIS , 2001 .

[40]  M. Sallum,et al.  Wing geometry of Anopheles darlingi Root (Diptera: Culicidae) in five major Brazilian ecoregions. , 2012, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[41]  R. Harbach Classification within the cosmopolitan genus Culex (Diptera: Culicidae): the foundation for molecular systematics and phylogenetic research. , 2011, Acta tropica.

[42]  J. Dujardin Morphometrics applied to medical entomology. , 2008, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[43]  C. Fruciano Measurement error in geometric morphometrics , 2016, Development Genes and Evolution.

[44]  Cláudia Torres Codeço,et al.  Spatial Evaluation and Modeling of Dengue Seroprevalence and Vector Density in Rio de Janeiro, Brazil , 2009, PLoS neglected tropical diseases.

[45]  Chwan-Chuen King,et al.  Effects of the El Niño-Southern Oscillation on dengue epidemics in Thailand, 1996-2005 , 2009, BMC public health.

[46]  J. F. Reinert List of abbreviations for currently valid generic-level taxa in family Culicidae (Diptera). , 2009 .

[47]  C. Klingenberg MorphoJ: an integrated software package for geometric morphometrics , 2011, Molecular ecology resources.

[48]  L. Kramer,et al.  West Nile virus vector competency of Culex quinquefasciatus mosquitoes in the Galapagos Islands. , 2011, The American journal of tropical medicine and hygiene.

[49]  I. Kitching,et al.  Reconsideration of anopheline mosquito phylogeny (Diptera: Culicidae: Anophelinae) based on morphological data , 2005 .