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Accurate species identification of horse flies (Diptera: Tabanidae) is crucial due to their role as vectors for various pathogens, which is essential for understanding their biology, devising strategies to control their populations, and enhancing disease surveillance. This study assessed the efficacy of outline-based geometric morphometrics (GM) by analyzing the wing cell contours of discal, first submarginal, and second submarginal cells to distinguish three morphologically similar Tabanus species commonly found in Thailand, T. megalops , T. rubidus , and T. striatus . Statistical analysis demonstrated significant size differences between T. rubidus and the two other species ( P  < 0.05), with T. rubidus exhibiting larger wing cells. Tabanus megalops and T. striatus had similar sizes; their size differences were not statistically significant. The accuracy of size analysis based on validated classification tests was relatively low, ranging from 64.67% to 68.67%. Nonetheless, all wing cell contours showed significant shape differences between the three species, as confirmed by Mahalanobis distance comparisons using 1000 permutation tests ( P  < 0.05). The shape of the first submarginal cell contour showed the highest classification accuracy (86.67%). Outline-based GM offers a significant advantage for analyzing fly specimens with incomplete wings that have intact cells. For damaged specimens, analyzing the contour of the first submarginal cell through this technique can be a viable alternative., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The blood-sucking flies of the genus Stomoxys Geoffroy, 1762 (Diptera: Muscidae) are significant ectoparasites that can cause irritation and transmit pathogens to both animals and humans. Within the genus Stomoxys , two species, Stomoxys bengalensis and Stomoxys sitiens , have similar morphology and coexist in the same habitat. Accurate species identification of these flies is crucial for understanding disease vectors and implementing effective control measures. In this study, we assessed the effectiveness of outline-based geometric morphometrics (GM) by analyzing the wing cell contour of the first posterior cell (R 5 ) to distinguish between species and sexes of S. bengalensis and S. sitiens . Our results demonstrate that the outline-based GM method is highly effective in distinguishing between species and sexes of these flies based on contour shape, with accuracy scores ranging from 90.0% to 97.5%. Therefore, outline-based GM emerges as a promising alternative to landmark-based GM or as a supplementary tool in conjunction with traditional morphology-based methods for species identification., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Accurate identification of mosquito species is critically important for monitoring and controlling the impact of human diseases they transmit. Here, we investigate four mosquito species: Aedes aegypti , Ae. albopictus , Ae. scutellaris and Verrallina dux that co-occur in tropical and subtropical regions, and whose morphological similarity challenges their accurate identification, a crucial requirement in entomological surveillance programs. Previous publications reveal a clear taxonomic signal embedded in wing cell landmark configuration, as well as in the external contour of the wings. We explored this signal for internal cells of the wings as well, to determine whether internal cells could uniformly provide the same taxonomic information. For each cell to be tentatively assigned to its respective species, i.e., to measure the amount of its taxonomic information, we used the shape of its contour, rather than its size. We show that (i) the taxonomic signal of wing shape is not uniformly spread among internal cells of the wing, and (ii) the amount of taxonomic information of a given cell depends on the species under comparison. This unequal taxonomic signal of internal cells is not related to size, nor to apparent shape complexity. The strong taxonomic signal of some cells ensures that even partly damaged wings can be used to improve species recognition.

Rhipicephalus microplus is one of the most widely distributed and economically important ticks able to transmit pathogens to vertebrate hosts which cause major constraints to public and livestock health. A better understanding of their population structure is crucial for the development of more effective control strategies. Modern morphometrics has enabled the quantification of size and shape variations to investigate population differences in anatomical structures. We therefore applied landmark-based and outline-based geometric morphometric approaches to study 85 field-collected adult females of three R. microplus Colombian populations along a distance of 28-67 km using the scutum and the basis capituli. Although size differences were found between some populations, mean shape and metric disparity of the analyzed structures were very similar, resulting in low classification scores (<41%). Thus, both structures support a metapopulation of R. microplus at a microgeographic level. The variance of shape between structures does not co-vary, likely indicating morphological modularity. These data provide first clues to understand the metric variation of R. microplus among natural populations from north-western Colombia., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This is the first study to explore the potential of various geometric morphometrics methods to help the morphological diagnostic of tsetse species, vectors of human and animal trypanosomiases in sub-Saharan Africa. We compared landmarks, semilandmarks and outlines techniques on male and female samples of species, and suggested adapted strategies according to the countries and their own Glossina fauna. We could compare up to 7 taxa belonging to the three main subgenera of the Glossina genus: Nemorhina (5 species), Glossina (1 species) and Austenina (1 species). Our sample included the major vectors of sleeping sickness: G. palpalis palpalis, G. p. gambiensis, G. fuscipes fuscipes and G. f. quanzensis, as well as two important vectors of African animal trypanosomoses: G. tachinoides and Glossina morsitans submorsitans. The average level of correct species recognition by the wing shape was satisfactory, and slightly higher for females than for males. The best scores of correct assignment, in both sexes, were obtained by the contour technique (96% of correct attribution in females, 92% in males), slightly higher than for semilandmarks (95% and 91%) or landmarks (94% and 89%) techniques. We made our images of wings freely available to be used as reference images (http://mome-clic.com), and we describe the conditions and the analytical steps to be followed to identify unknown specimens using external reference images. Under adequate conditions, such use of reference images obtained from a free access server could help species identification of new samples anywhere in Africa., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Adult flies of the genus Stomoxys Geoffroy, 1762 (Diptera: Muscidae), especially S. pullus Austen, 1909, S. uruma Shinonaga et Kano, 1966 and S. indicus Picard, 1908, are morphologically similar and sometimes difficult to distinguish when using external morphological characteristics. These species may act as vectors and/or potential vectors of many pathogens (virus, bacteria and protozoa). Their correct identification is important to target the vectors involved in the transmission of the pathogens and also helps in the fly control program.The aim of the present study was to distinguish three species which are difficult to separate using traditional diagnostic characters for species of Stomoxys such as colour patterns and body proportions. Modern morphometrics, both landmark and outline-based, was used to access wing geometry of S. pullus, S. uruma and S. indicus. A total of 198 and 190 wing pictures were analysed for landmark- and outline-based approaches, respectively. Wing shape was able to separate species and sexes of the three Stomoxys flies with highly significant difference of Mahalanobis distances. The cross-validated classification scores ranged from 76% to 100% for landmark and 77% to 96% for outline-based morphometrics. The geometry of wing features appears to be a very useful, low-cost tool to distinguish among the vectors S. pullus, S. uruma and S. indicus.

Aedes aegypti (Diptera: Culicidae) (L.), Ae. albopictus (Skuse), and Ae. scutellaris (Walker) are important mosquito vectors of dengue and chikungunya viruses. They are morphologically similar and sympatric in some parts of their distribution; therefore, there is a risk of incorrect morphological identification. Any confusion could have a negative impact on epidemiological studies or control strategies. Therefore, we explored two modern tools to supplement current morphological identification: DNA barcoding and geometric morphometric analyses. Field larvae were reared to adults and carefully classified based on morphological traits. The genetic analysis was based on the 658bp each of 30COI sequences. Some Culex spp., Mansonia bonneae, were included as outgroups, and inclusion of a few other Aedes spp. facilitated phylogenetic inference of the relationship between Ae. albopictus and Ae. scutellaris. The two species were separated by an average interspecific divergence of 0.123 (0.119-0.127). Morphometric examination included landmark- (392 specimens) and outline-based (317 specimens) techniques. The shape of the wing showed different discriminating power based on sex and digitizing technique. This is the first time that Ae. scutellaris and Ae. albopictus have been compared using these two techniques. We confirm that these morphologically close species are valid, and that geometric morphometrics can considerably increase the reliability of morphological identification., (Copyright © 2016 Elsevier B.V. All rights reserved.)

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., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Using outlines derived from a widely used set of line drawings, we created stimuli geared towards the investigation of contour integration and texture segmentation using shapes of everyday objects. Each stimulus consisted of Gabor elements positioned and oriented curvilinearly along the outline of an object, embedded within a larger Gabor array of homogeneous density. We created six versions of the resulting Gaborized outline stimuli by varying the orientations of elements inside and outside the outline. Data from two experiments, in which participants attempted to identify the objects in the stimuli, provide norms for identifiability and name agreement, and show differences in identifiability between stimulus versions. While there was substantial variability between the individual objects in our stimulus set, further analyses suggest a number of stimulus properties which are generally predictive of identification performance. The stimuli and the accompanying normative data, both available on our website (http://www.gestaltrevision.be/sources/gaboroutlines), provide a useful tool to further investigate contour integration and texture segmentation in both normal and clinical populations, especially when top-down influences on these processes, such as the role of prior knowledge of familiar objects, are of main interest.