Geometric morphometrics for the taxonomy of 11 species of Anopheles (Nyssorhynchus) mosquitoes.

The subgenus Anopheles (Nyssorhynchus) (Diptera: Culicidae) includes the primary vectors of Plasmodium spp. in Colombia. Most adult females of this subgenus are difficult to identify in the field using the available keys. With the objective of further investigating the discriminatory power of modern morphometrics, both landmark-based and outline-based approaches were explored using the wing venation geometry of 11 Anopheles (Nyssorhynchus) species. Wing shape was able to separate the closest species of the subgenus. When the 11 species were analysed together, validated classification scores on average 5.3-8.6 times higher than those expected by chance were observed. These scores computed from the total sample of 11 species were not satisfactory for the recognition of Anopheles benarrochi B, Anopheles oswaldoi s.l. and Anopheles strodei. These sibling species were captured in sympatry. To improve the identification power of the morphometric tool, it was necessary to analyse these species separately from the remaining species. The best classification scores were obtained using a combination of 12 landmarks collected not only on the intersections of wing veins, but also on spots. An outline approach also gave excellent reclassification scores. Another pair of sibling species, collected in allopatry, Anopheles nuneztovari and Anopheles rangeli, also showed high classification scores.

Outline-based morphometrics, an overlooked method in arthropod studies?

Modern methods allow a geometric representation of forms, separating size and shape. In entomology, as well as in many other fields involving arthropod studies, shape variation has proved useful for species identification and population characterization. In medical entomology, it has been applied to very specific questions such as population structure, reinfestation of insecticide-treated areas and cryptic species recognition. For shape comparisons, great importance is given to the quality of landmarks in terms of comparability. Two conceptually and statistically separate approaches are: (i) landmark-based morphometrics, based on the relative position of a few anatomical "true" or "traditional" landmarks, and (ii) outline-based morphometrics, which captures the contour of forms through a sequence of close "pseudo-landmarks". Most of the studies on insects of medical, veterinary or economic importance make use of the landmark approach. The present survey makes a case for the outline method, here based on elliptic Fourier analysis. The collection of pseudo-landmarks may require the manual digitization of many points and, for this reason, might appear less attractive. It, however, has the ability to compare homologous organs or structures having no landmarks at all. This strength offers the possibility to study a wider range of anatomical structures and thus, a larger range of arthropods. We present a few examples highlighting its interest for separating close or cryptic species, or characterizing conspecific geographic populations, in a series of different vector organisms. In this simple application, i.e. the recognition of close or cryptic forms, the outline approach provided similar scores as those obtained by the landmark-based approach.

Development of a geographical distribution model of Rhodnius pallescens Barber, 1932 using environmental data recorded by remote sensing.

Rhodnius pallescens, main vector of Trypanosoma cruzi in Panama and secondary vector in Colombia, Costa Rica and Nicaragua, represents an important epidemiological risk in those countries. It occupies sylvatic ecotopes, and because of this its distribution and abundance could be conditioned by environmental factors. In this work, we integrated environmental variables recorded by remote sensing and data of R. pallescens presence in the countries mentioned above in order to know the environmental variables with better capacity to describe the insects' distribution, which will help to entomological surveillance and control. Standard discriminant analysis (DA) was used to determine if there is a significant difference in the environmental variation between the presence and the absence sites of R. pallescens. Forward stepwise discriminant analysis (fDA) was used to determine the environmental variables which better discriminated between presence and absence sites, and to construct a predictive map of geographical distribution. Univariate analyses were used to determine the discriminatory power of individual variables. The model derived from DA showed 89% of sensitivity and 92.8% of specificity. Multivariate and univariate analyses showed the vapor pressure deficit minimum as the principal variable among the nine most important to describe the distribution of the species, which is in agreement with the R. pallescens stenohydric status. Map shows insects' distribution predicted by environmental variables, and moreover includes the distribution of most species belonging to Rhodnius genus, except R. domesticus, R. nasutus and R. neglectus. We interpreted these results as a reflection of the common evolution of the most Rhodnius species, except for the last ones that probably evolved isolated due to particular environmental conditions. In conclusion, this study showed that a reduced number of environmental variables can predict the distribution of R. pallescens and related species. This methodology can be very useful to make critical decisions for vectorial surveillance and control of Chagas disease vectors.