Discrimination of four Culex (Culex) species from the Neotropics based on geometric morphometrics

Achieving correct identification of Culex species is difficult because many anatomical characters of larvae and females are polymorphic or overlap among distinct species. The overlapping is known to occur between Culex bidens, Cx. interfor, Cx. mollis and Cx. tatoi, from the subgenus Culex. The first three were incriminated in viruses’ transmission in Argentina. The purpose was to distinguish between specimens of four species using geometric morphometric procedures. From field and entomological collections, 10 type I and 10 type II landmarks on the wings of females and the dorsomentum of larvae, respectively, were defined. The free morphometric software modules by J.P. Dujardin were used. Landmark coordinates were submitted to Procrustes and TPS analyses to generate size (centroid size, CS) and shape (partial warps, PW) variables. Size analysis was performed by nonparametric comparisons of CS measurements based on permutations, and shape, by submission of PW to discriminant analysis. Re-identification and identification of external specimens were also realized. Wing and Dm shapes gave similar results. The individuals arranged into three groups coinciding with Cx. bidens + Cx. interfor, Cx. mollis and Cx. tatoi. The latter two had a very different wing and Dm shapes. The accurate re-identification was greater than or equal to 77 % in all cases, and the identification of external specimens was achieved for all the species and both structures. Larval characters are more informative than female features. Geometric morphometrics, as a complementary tool, will facilitate identification in junction with others, such as DNA sequence analysis.

[1]  R. A. Bram Classification of Culex subgenus Culex in the New World (Díptera: Culicidae). , 1967 .

[2]  R. Cruickshank,et al.  The seven deadly sins of DNA barcoding , 2012, Molecular ecology resources.

[3]  M. Correa,et al.  Wing geometric morphometrics and molecular assessment of members in the Albitarsis Complex from Colombia , 2013, Molecular ecology resources.

[4]  J. N. Belkin The Mosquitoes of the South Pacific (Diptera, Culicidae). , 1962 .

[5]  Professor Dr. Friedrich Ruttner Biogeography and Taxonomy of Honeybees , 1987, Springer Berlin Heidelberg.

[6]  G. Dobigny,et al.  Geometric morphometrics, neural networks and diagnosis of sibling Taterillus species (Rodentia, Gerbillinae) , 2002 .

[7]  B. Sharp,et al.  Temperature‐dependent variation in Anopheles merus larval head capsule width and adult wing length: implications for anopheline taxonomy , 1991, Medical and veterinary entomology.

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

[9]  F. James Rohlf,et al.  Geometric morphometrics and phylogeny , 2000 .

[10]  M. Baylac,et al.  Elliptic Fourier analysis of the form of genitalia in two Spodoptera species and their hybrids (Lepidoptera: Noctuidae) , 2001 .

[11]  C. Villemant,et al.  Combining geometric morphometrics with pattern recognition for the investigation of species complexes , 2003 .

[12]  P. Good,et al.  Permutation Tests: A Practical Guide to Resampling Methods for Testing Hypotheses , 1995 .

[13]  A. Tenório,et al.  Cocirculation of Rio Negro Virus (RNV) and Pixuna Virus (PIXV) in Tucumán province, Argentina , 2010, Tropical medicine & international health.

[14]  Wing geometry of Culex coronator (Diptera: Culicidae) from South and Southeast Brazil , 2014, Parasites & Vectors.

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

[16]  J. Dujardin,et al.  Chromosomal and environmental determinants of morphometric variation in natural populations of the malaria vector Anopheles funestus in Cameroon. , 2011, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[17]  J. Dujardin,et al.  The geometry of the wing of Aedes (Stegomyia) aegypti in isofemale lines through successive generations. , 2008, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[18]  J. Dujardin,et al.  Geographical versus interspecific differentiation of sand flies (Diptera: Psychodidae): a landmark data analysis , 2003, Bulletin of Entomological Research.

[19]  R. Harbach,et al.  The Culicidae (Diptera): a review of taxonomy, classification and phylogeny* , 2007 .

[20]  J. Navarro,et al.  Phylogenetic relationships among eighteen neotropical Culicini species. , 2000, Journal of the American Mosquito Control Association.

[21]  H. G. Dyar The mosquitoes of the Americas , 1928 .

[22]  J. Dujardin,et al.  Influence of larval density or food variation on the geometry of the wing of Aedes (Stegomyia) aegypti , 2007, Tropical medicine & international health : TM & IH.

[23]  F. Rohlf,et al.  Extensions of the Procrustes Method for the Optimal Superimposition of Landmarks , 1990 .

[24]  H. G. Dyar,et al.  Notes on some American mosquitoes with descriptions of new species , 1906 .

[25]  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.

[26]  F. Rohlf,et al.  A revolution morphometrics. , 1993, Trends in ecology & evolution.

[27]  K. Spitze Population structure in Daphnia obtusa: quantitative genetic and allozymic variation. , 1993, Genetics.

[28]  D. Kendall MORPHOMETRIC TOOLS FOR LANDMARK DATA: GEOMETRY AND BIOLOGY , 1994 .

[29]  R. Harbach,et al.  Recognition of Culex bidens Dyar and Culex interfor Dyar (Diptera: Culicidae) as separate species. , 1986 .

[30]  W. Almirón,et al.  Culex (Culex) interfor Dyar (Diptera: Culicidae), morphological description including previously unknown life stages , 1996 .