Wing morphology analysis of Anopheles mosquitoes using scanning electron microscopy and minkowski functionals for species distinction

Abstract Background Digital image processing has become an essential tool for species identification and surface characterization. Among advanced morphological analyses, Minkowski functionals (MFs), a mathematical descriptor derived from integral geometry, offers a quantitative approach to assessing surface features. This study focuses on using scanning electron microscopy (SEM) combined with MFs to analyze the wing morphology of Anopheles mosquitoes, which are significant vectors of malaria. Understanding fine-scale wing morphology is critical for improving species identification and developing effective disease control strategies. Results SEM analysis revealed morphological differences between the two species on both dorsal and ventral wing surfaces. It was observed that there was the presence of scales along the veins and edges of the wings and as well as long hairs distributed across the wing area. High-magnification images enabled detailed analysis of nanometric structures. Quantitative analysis using MFs indicated that An. aquasalis wings presented more pronounced surface elevations, greater height variation, and a higher density of peaks and valleys, while An. darlingi exhibited smoother and more uniform surfaces and presence of nanostructures with the presence of nanostructures. These functional analyses provided a comprehensive understanding of the differences in surface roughness and structural connectivity between the two species. Conclusions The combination of SEM and MFs proved effective for distinguishing mosquito species based on wing surface architecture. This high-resolution, quantitative approach serves as a valuable complement to identification, enabling precise species distinction, and enhancing the understanding of the morphological characteristics that influence flight dynamics, adaptation, dispersion and vector capacity of mosquitoes, contributing to better disease control strategies and potential applications in various biotechnical fields. The application of MFs in conjunction with SEM provides a robust method for quantifying complex surface morphology and can be expanded to other entomological and biological studies.