Unveiling the Wing Shape Variation in Northern Altiplano Ecosystems: The Example of the Butterfly Phulia nymphula Using Geometric Morphometrics.

The Andean Altiplano, characterized by its extreme climatic conditions and high levels of biodiversity, provides a unique environment for studying ecological and evolutionary adaptations in insect morphology. Butterflies, due their large wing surface compared to body surface, and wide distribution among a geographical area given the flight capabilities provided by their wings, constitute a good biological model to study morphological adaptations following extreme weathers. This study focuses on Phulia nymphula, a butterfly species widely distributed in the Andes, to evaluate wing shape variation across six localities in the Northern Chilean Altiplano. The geometric morphometrics analysis of 77 specimens from six locations from the Chilean Altiplano (Caquena, Sorapata Lake, Chungará, Casiri Macho Lake, Surire Salt Flat, and Visviri) revealed significant differences in wing shape among populations. According to the presented results, variations are likely influenced by local environmental conditions and selective pressures, suggesting specific adaptations to the microhabitats of the Altiplano. The first three principal components represented 60.92% of the total wing shape variation. The detected morphological differences indicate adaptive divergence among populations, reflecting evolutionary responses to the extreme and fragmented conditions of the Altiplano. This study gives insights into the understanding of how high-altitude species can diversify and adapt through morphological variation, providing evidence of ecological and evolutionary processes shaping biodiversity in extreme environments.

Deciphering the Hearts: Geometric Morphometrics Reveals Shape Variation in Abatus Sea Urchins across Subantarctic and Antarctic Seas.

Abatus is a genus of irregular brooding sea urchins to the Southern Ocean. Among the 11 described species, three shared morphological traits and present an infaunal lifestyle in the infralittoral from the Subantarctic province; A. cavernosus in Patagonia, A. cordatus in Kerguelen, and A. agassizii in Tierra del Fuego and South Shetlands. The systematic of Abatus, based on morphological characters and incomplete phylogenies, is complex and largely unresolved. This study evaluates the shape variation among these species using geometric morphometrics analysis (GM). For this, 72 individuals from four locations; South Shetlands, Kerguelen, Patagonia, and Falklands/Malvinas were photographed, and 37 landmarks were digitized. To evaluate the shape differences among species, a principal component analysis and a Procrustes ANOVA were performed. Our results showed a marked difference between the Falklands/Malvinas and the other localities, characterized by a narrower and more elongated shape and a significant influence of location in shape but not sex. Additionally, the effect of allometry was evaluated using a permutation test and a regression between shape and size, showing significant shape changes during growth in all groups. The possibility that the Falklands/Malvinas group shows phenotypic plasticity or represents a distinct evolutionary unit is discussed. Finally, GM proved to be a powerful tool to differentiate these species, highlighting its utility in systematic studies.

Breaking the Law: Is It Correct to Use the Converse Bergmann Rule in Ceroglossus chilensis? An Overview Using Geometric Morphometrics.

The converse Bergmann's rule is a pattern of body size variation observed in many ectothermic organisms that contradicts the classic Bergmann's rule and suggests that individuals inhabiting warmer climates tend to exhibit larger body sizes compared to those inhabiting colder environments. Due to the thermoregulatory nature of Bergmann's rule, its application among ectotherms might prove to be more complicated, given that these organisms obtain heat by absorbing it from their habitat. The existence of this inverse pattern therefore challenges the prevailing notion that larger body size is universally advantageous in colder climates. Ceroglossus chilensis is a native Chilean beetle that has the largest latitudinal range of any species in the genus, from 34.3° S to 47.8° S. Within Chile, it continuously inhabits regions extending from Maule to Aysen, thriving on both native and non-native forest species. Beyond their remarkable color variation, populations of C. chilensis show minimal morphological disparity, noticeable only through advanced morphological techniques (geometric morphometrics). Based on both (1) the "temperature-size rule", which suggests that body size decreases with increasing temperature, and (2) the reduced resource availability in high-latitude environments that may lead to smaller body sizes, we predict that C. chilensis populations will follow the converse Bergmann's rule. Our results show a clear converse pattern to the normal Bergmann rule, where smaller centroid sizes were found to be measured in the specimens inhabiting the southern areas of Chile. Understanding the prevalence of the converse Bergmann's rule for ectotherm animals and how often this rule is broken is of utmost importance to understand the underlying mechanisms allowing organisms to adapt to different environments and the selective pressures they face.

Unraveling biotypes of the northern house mosquito, Culex pipiens s.l. (Diptera: Culicidae): molecular differentiation and morphometric analysis.

Geometric morphometrics was used to determine whether geographic isolation could explain differences in wing size and shape between and within continental (27°S to 41°S) and insular (Rapa Nui) populations of Culex pipiens s.s. Linnaeus and their biotypes (f. pipiens and f. molestus). Molecular protocols based on polymorphisms in the second intron of nuclear locus ace-2 (acetylcholinesterase-2) were used to differentiate Cx. pipiens s.s. from Cx. quinquefasciatus Say, and an assay based on polymorphisms in the flanking region of a microsatellite locus (CQ11) was used to identify biotypes. Culex pipiens f. molestus and hybrids shared larval habitats in all continental sites, while Cx. pipiens f. pipiens was found in 5 of the 10 sites. Only biotype molestus was found in Rapa Nui (Easter Island) Pipiens and molestus biotypes occur sympatrically in aboveground locations, and only molestus was found in the underground site (ME). Biotype molestus was dominant in rural locations and preferably anthropophilic. These results agree with the ecological descriptions previously reported for the biotypes of Cx. pipiens s.s. Procrustes ANOVA only showed differences in centroid size between biotypes in females and males and did not show significant differences in wing shape. However, we found significant differences among the geographic areas in the centroid size and wing shape of both females and males. Particularly, the population of Rapa Nui Island had shorter wings than the continental populations. The results highlight the effects of geographic and environmental processes on morphotypes in vector mosquitoes.

Unraveling the Sexual Dimorphism of First Instar Nymphs of the Giant Stick Insect, Cladomorphus phyllinus Gray, 1835, from the Atlantic Forest, Brazil.

The first instar nymphs, both male and female, of the giant stick insect Cladomorphus phyllinus Gray, 1835 were carefully described and measured, revealing a remarkable sexual dimorphism that is considered rare among insects and is poorly explored in the order Phasmida. The studied F1 nymphs originated in captivity from eggs laid by a coupled female specimen collected in the Atlantic Forest in the vicinity of Petrópolis city, state of Rio de Janeiro, Brazil. The first instar nymphs of C. phyllinus were measured and illustrated in high-resolution photographs to show the general aspects and details of sexually dimorphic traits, making clear the phenotypic differences in the sexes. A total of 100 nymphs were kept alive until morphological sexual dimorphism was confirmed and quantified. All recently hatched first instar nymphs were separated based on the presumed male and female characteristics, i.e., the presence and absence of the suture in the metanotum in the males and females, respectively, had their sexes confirmed in 100% of the specimens as previously assigned. These results confirm this new morphological trait, which here is named "alar suture" as sex-specific in the first instar nymphs, a novelty in this stage of development of sexual differentiation. In addition, the distinct conformations of the last three abdominal sternites of both sexes were recorded.

Bergmann's Rule under Rocks: Testing the Influence of Latitude and Temperature on a Chiton from Mexican Marine Ecoregions.

Bergmann's rule relates the trend of increasing body size with higher latitudes, where colder climates are found. In the Mexican Pacific, three marine ecoregions are distinguishable across a latitudinal gradient. Stenoplax limaciformis is an abundant chiton species that is distributed on rocky shores in these ecoregions. Geometric morphometric analyses were performed to describe the shape and size variation of S. limaciformis between marine ecoregions that vary in sea surface temperature with latitude, thus testing Bergmann's rule. Individuals' body shape ranged from elongated to wide bodies. Although there was variation in chitons' body shape and size, the was no evidence of allometry among localities. The Gulf of California is the northernmost ecoregion evaluated in this work, where larger chitons were observed and lower sea surface temperature values were registered. The results suggest that S. limaciformis follows a trend to Bergmann's rule, such as endotherms. These mollusks do not need heat dissipation, but they do need to retain moisture. In addition, larger chitons were observed in zones with high primary productivity, suggesting that chitons do not delay their maturation due to food shortage.

Description of Bothriurus mistral n. sp., the highest-dwelling Bothriurus from the western Andes (Scorpiones, Bothriuridae), using multiple morphometric approaches.

We describe Bothriurus mistral n. sp. (Scorpiones, Bothriuridae) from the Chilean north-central Andes of the Coquimbo Region. This is the highest elevational discovery for Bothriurus in the western slopes of the Andes. This species was collected in the Estero Derecho Private Protected Area and Natural Sanctuary as part of the First National Biodiversity Inventory of Chile of the Integrated System for Monitoring and Evaluation of Native Forest Ecosystems (SIMEF). Bothriurus mistral n. sp. is closely related to Bothriurus coriaceus Pocock, 1893, from the lowlands of central Chile. This integrative research includes a combination of traditional morphometrics and geometric morphometric analyses to support the taxonomic delimitation of the species.

Unraveling the morphological patterns of a subantarctic eelpout: a geometric morphometric approach.

Phenotypic variation in organisms depends on the genotype and the environmental constraints of the habitat that they exploit. Therefore, for marine species inhabiting contrasting aquatic conditions, it is expected to find covariation between the shape and its spatial distribution. We studied the morphology of the head and cephalic sensory canals of the eelpout Austrolycus depressiceps (4.5-22.5 cm TL) across its latitudinal distribution in South Pacific (45°S-55°S). Geometric morphometric analyses show that the shape varied from individuals with larger snout and an extended suborbital canal to individuals with shorter snouts and frontally compressed suborbital canal. There was size variation across the sampled populations, but that size does not have a clear latitudinal gradient. Only 8% of the shape variation relates to this size variation (allometry), represented by a decrease in the relative size of the eye, and a depression of the posterior margin of the head. There were spatial differences in the shape of the head, but these differences were probably caused by allometric effects. Additionally, 2 of the canals of the cephalic lateralis pores and the head shape showed modularity in its development. This study shows that the morphology of marine fish with a shallow distribution varies across distances of hundreds of kilometers (i.e., phenotypic modulation).

Ecomorphology and Morphological Disparity of Caquetaia Kraussii (Perciformes: Cichlidae) in Colombia.

Understanding the interspecific morphological variability of Caquetaia kraussii (Perciformes: Cichlidae) between different localities in its distribution range is becoming essential, as this species constitutes a valuable resource for the economy and subsistence of the local human communities where it is endemic in Colombia and Venezuela. In order to develop efficient farming and handling plans for this species, a deep understanding of the factors and mechanisms generating morphological variability is crucial. This study analyzes the morphological variability of C. kraussii by using geometric morphometrics in four localities distributed between the Dique and North channels, which are part of the Bolívar department in Colombia. Likewise, the effect of environmental variables such as temperature (T°), dissolved oxygen (OD) and pH on morphological variability was analyzed using a partial least squares approach. The results show that environmental stress has an influence on ~10% of the body shape of C. kraussii, whereas ~90% of the body shape is not directly influenced by environmental parameters, suggesting an effect from stress related to sexual dimorphism. Similarly, the analyses show shape variation among localities, mainly between populations of lotic environments and those of lentic environments. This morphological disparity seems to be subject to environmental and sexual stresses in the different localities.

Evolvability in the Cephalothoracic Structural Complexity of Aegla araucaniensis (Crustacea: Decapoda) Determined by a Developmental System with Low Covariational Constraint.

The integration of complex structures is proportional to the intensity of the structural fusion; its consequences are better known than the covariational effects under less restrictive mechanisms. The synthesis of a palimpsest model based on two early parallel pathways and a later direct pathway explains the cephalothoracic complexity of decapod crustaceans. Using this model, we tested the evolvability of the developmental modularity in Aegla araucaniensis, an anomuran crab with an evident adaptive sexual dimorphism. The asymmetric patterns found on the landmark configurations suggest independent perturbations of the parallel pathways in each module and a stable asymmetry variance near the fusion by canalization of the direct pathway, which was more intense in males. The greater covariational flexibility imposed by the parallel pathways promotes the expression of gonadic modularity that favors the reproductive output in females and agonistic modularity that contributes to mating success in males. Under these divergent expressions of evolvability, the smaller difference between developmental modularity and agonistic modularity in males suggests higher levels of canalization due to a relatively more intense structural fusion. We conclude that: (1) the cephalothorax of A. araucaniensis is an evolvable structure, where parallel pathways promote sexual disruptions in the expressions of functional modularity, which are more restricted in males, and (2) the cephalothoracic palimpsest of decapods has empirical advantages in studying the developmental causes of evolution of complex structures.

Morphological Stasis in Time? A Triatoma brasiliensis brasiliensis Study Using Geometric Morphometrics in the Long Run.

Triatoma brasiliensis brasiliensis Neiva, 1911 is one of the most important vectors of Chagas disease in the Brazilian semiarid regions in the north-east. The risk imposed by T. b. brasiliensis to the human populations, due to frequent invasions and/or colonization of the domiciles, demands constant monitoring and control actions as well as an understanding of its evolutionary process. In this context, the following research studies the pattern of shape adaptation over time using a large dataset from 102 years of specimen collections in order to identify the morphological plasticity of this vector in Brazil. This dataset was analyzed using geometric morphometrics tools and the timescale was divided into eight different groups, containing specimens from 1912 to 2014. Geometric morphometrics analysis showed an interesting morphological stasis in the wing shape of T. b. brasiliensis, which allowed us to understand the high capacity of adaptation to changes in climate condition through time, and the invasive status which Triatoma species have around the world. Moreover, these results showed novel findings as an interesting phenotypic pattern, with no modifications in more than 100 years, leading us to understand the shape evolution in Triatominae as a vector species of diseases.

Genetics, Morphometrics and Health Characterization of Green Turtle Foraging Grounds in Mainland and Insular Chile.

Two divergent genetic lineages have been described for the endangered green turtle in the Pacific Ocean, occurring sympatrically in some foraging grounds. Chile has seven known green turtle foraging grounds, hosting mainly juveniles of different lineages. Unfortunately, anthropic factors have led to the decline or disappearance of most foraging aggregations. We investigated age-class/sex structure, morphological variation, genetic diversity and structure, and health status of turtles from two mainland (Bahia Salado and Playa Chinchorro) and one insular (Easter Island) Chilean foraging grounds. Bahia Salado is composed of juveniles, and with Playa Chinchorro, exclusively harbors individuals of the north-central/eastern Pacific lineage, with Galapagos as the major genetic contributor. Conversely, Easter Island hosts juveniles and adults from both the eastern Pacific and French Polynesia. Morphological variation was found between lineages and foraging grounds, suggesting an underlying genetic component but also an environmental influence. Turtles from Easter Island, unlike Bahia Salado, exhibited injuries/alterations probably related to anthropic threats. Our findings point to establishing legal protection for mainland Chile's foraging grounds, and to ensure that the administrative plan for Easter Island's marine protected area maintains ecosystem health, turtle population viability, and related cultural and touristic activities.

Insularity and Aridity as Drivers of Mandibular Disparity in Thylamys elegans (Waterhouse, 1839) from Populations of the Atacama Desert, Chile.

Island ecosystems differ in several elements from mainland ecosystems and may induce variations related to natural selection and patterns of adaptation in most aspects of the biology of an organism. Thylamys elegans (Waterhouse, 1839) is a marsupial endemic to Chile, distributed from Loa River to Concepción. Historically, three subspecies have been described: Thylamys elegans elegans, Thylamys elegans coquimbensis and Thylamys elegans soricinus. For this research, two morphometric approaches and a biomechanical model were used to compare the mandible shapes and biomechanics between two Chilean mouse opossum populations belonging to different subspecies: one from the coastal desert of Chile (T. e. coquimbensis) and the other from the central inland region (T. e. elegans). Additionally, mandibles of insular populations found in the Reserva Nacional Pinguino de Humboldt (RNPH)), from which the subspecies association is unknown, were also included. The results showed that insular populations have differences in mandibular shapes, sizes and biomechanical characteristics compared to continental populations, which may be related to environmental variables like aridity and vegetation cover, prey type, insularity effects and/or the founder effect on micromammals, apart from vicariance hypotheses and other selective pressures.

Drosophila Wing Integration and Modularity: A Multi-Level Approach to Understand the History of Morphological Structures.

Static, developmental, and evolutionary variation are different sources of morphological variation which can be quantified using morphometrics tools. In the present study we have carried out a comparative multiple level study of integration (i.e., static, developmental, and evolutionary) to acquire insight about the relationships that exist between different integration levels, as well as to better understand their involvement in the evolutionary processes related to the diversification of Drosophila's wing shape. This approach was applied to analyse wing evolution in 59 species across the whole genus in a large dataset (~10,000 wings were studied). Static integration was analysed using principal component analysis, thus providing an integration measurement for overall wing shape. Developmental integration was studied between wing parts by using a partial least squares method between the anterior and posterior compartments of the wing. Evolutionary integration was analysed using independent contrasts. The present results show that all Drosophila species exhibit strong morphological integration at different levels. The strong integration and overall similarities observed at multiple integration levels suggest a shared mechanism underlying this variation, which could result as consequence of genetic drift acting on the wing shape of Drosophila.

Green, yellow or black? Genetic differentiation and adaptation signatures in a highly migratory marine turtle.

Marine species may exhibit genetic structure accompanied by phenotypic differentiation related to adaptation despite their high mobility. Two shape-based morphotypes have been identified for the green turtle (Chelonia mydas) in the Pacific Ocean: the south-central/western or yellow turtle and north-central/eastern or black turtle. The genetic differentiation between these morphotypes and the adaptation of the black turtle to environmentally contrasting conditions of the eastern Pacific region has remained a mystery for decades. Here we addressed both questions using a reduced-representation genome approach (Dartseq; 9473 neutral SNPs) and identifying candidate outlier loci (67 outlier SNPs) of biological relevance between shape-based morphotypes from eight Pacific foraging grounds (n = 158). Our results support genetic divergence between morphotypes, probably arising from strong natal homing behaviour. Genes and enriched biological functions linked to thermoregulation, hypoxia, melanism, morphogenesis, osmoregulation, diet and reproduction were found to be outliers for differentiation, providing evidence for adaptation of C. mydas to the eastern Pacific region and suggesting independent evolutionary trajectories of the shape-based morphotypes. Our findings support the evolutionary distinctness of the enigmatic black turtle and contribute to the adaptive research and conservation genomics of a long-lived and highly mobile vertebrate.

Unraveling the Morphological Variation of Triatoma infestans in the Peridomestic Habitats of Chuquisaca Bolivia: A Geometric Morphometric Approach.

Morphometrics has been used on Triatomines, a well-known phenotypically variable insect, to understand the process of morphological plasticity and infer the changes of this phenomenon. The following research was carried out in two regions of the inter-Andean valleys and two Chaco regions of Chuquisaca-Bolivia. Triatoma infestans adults were collected from the peridomestic (pens and chicken coops) along a geographic gradient in order to evaluate the morphological differentiation between groups and their pattern of sexual shape dimorphism. Geometric morphometric methods were applied on the wings and heads of T. infestans. The main findings include that we proved sexual dimorphism in heads and wings, determined the impact of environmental factors on size and shape and validated the impact of nutrition on head shape variation. These results show that geometric morphometric procedures can be used to provide key insight into the biological adaptation of T. infestans on different biotic (nutrition) and abiotic (environment) conditions, which could serve in understanding and evaluating infestation processes and further vector control programs.

Morfometría geométrica y sus nuevas aplicaciones en ecología y biología evolutiva: Parte 2 / Geometric morphometric and its new applications in ecology and evolutionary biology: Part 2

RESUMEN: La presente revisión entrega una visión actualizada del estudio de la morfometría geométrica y sus aplicaciones más actuales en ecología y biología evolutiva, metodología con una amplia variación en los últimos 5 años de su primera versión en International Journal of Morphology. La Morfometría geométrica es una herramienta que permite evaluar las variaciones morfológicas con factores subyacentes, siendo una herramienta más sensible que la morfometría tradicional, lo que permite detectar mínimos cambios de variación morfológica. Lo que la ha vuelto una herramienta notable para responder preguntas de biología comparada centradas en caracteres anatómicos. En sus comienzos fue una herramienta usada principalmente para responder preguntas taxonómicas, y para diferenciar a nivel de individuos, poblaciones o especies. No obstante, en los últimos años la cantidad de preguntas y problemáticas en las que se aplica, ha diversificado considerablemente, pasando a ser una herramienta muy precisa para responder preguntas de variación morfológica en contextos ecológicos y evolutivos. Ya ha pasado casi media década desde la última revisión del método, por lo que éste trabajo tiene como objetivo analizar los cambios metodológicos y los nuevos enfoques usados en Morfometría geométrica, y presentar una pequeña guía introductoria a éstos nuevos métodos, sus usos y aplicaciones.

SUMMARY: This study provides an updated vision of the study of Geometric Morphometrics and its most recent application in ecology and evolutionary biology, covering a wide variation in methodology occurring in the last 5 years since the first version published in the International Journal of Morphology. Geometric Morphometrics is a tool that allows evaluating morphological variations with underlying factors, with a higher sensitivity than traditional morphology, so that minimum changes of morphological variation can be detected. Therefore, it has turned into an outstanding tool to answer questions of comparative biology focused on anatomic characters. At the beginning, it was a tool mainly used to answer taxonomic questions and for differentiation at individual, population or species level. However, in the last years, the number of questions and problematic on which it is applied, has diversified considerably, turning it into a very accurate tool to answer questions of morphological variation in ecologic and evolutionary contexts. Almost half a decade has elapsed since the last revision of the method, so this work is intended to analyze the methodological changes and the new approaches used in Geometric Morphometrics, including a brief introductory guideline to these new methods, their uses and applications.

Measuring the Inter and Intraspecific Sexual Shape Dimorphism and Body Shape Variation in Generalist Ground Beetles in Russia.

Ground beetles in multiple species vary greatly in the expression of the shape on sexual traits, resulting in a sexual shape dimorphism as a consequence of sexual selection differences. The present research focuses on the study of inter and intrasexual sexual shape dimorphism of two generalist genera of ground beetles Pterostichus and Carabus. Geometric morphometric methods were applied to five generalist species of ground beetles Carabus exaratus, C. granulatus, Pterostichus melanarius, P. niger, and P. oblongopunctatus and several multivariate analyses were applied for two different traits, abdomen and elytra. Three of the five species analyzed showed high levels of sex-based shape dimorphism. However, the most generalist species, P. melanarius and P. oblongopunctatus, did not evidence shape-based sexual dimorphism differentiation in both of the analyzed traits, as statistically confirmed based on the permutation of pairwise comparison of the Mahalanobis distances of a sex-species classifier. It is generally known that environmental stress in natural populations can affect the fitness expression, principally related to sexual fecundity, being that this pattern is more evident in non-generalist species. In our results, the contrary pattern was found, with the absence of sexual shape dimorphism for two of the three generalist species analyzed. On the other hand, the interspecies shape variation was clearly identified using principal component analysis of both of the analyzed traits. Finally, this research is the first to analyze the relationship between sexual shape dimorphism in Russian ground beetles, evidencing the lack of understanding of the mechanism underlying the sexual dimorphism, especially in species living in extreme environments.

Assessment of Shape Variation Patterns in Triatoma infestans (Klug 1834) (Hemiptera: Reduviidae: Triatominae): A First Report in Populations from Bolivia.

The morphological variations of four populations of geographically isolated Triatoma infestans located in the area of inter-Andean valleys and Chaco of Chuquisaca, Bolivia, were evaluated. Fifty-three females and sixty-one males were collected in the peri-domicile and analyzed with geometric morphometrics tools to study the patterns of the head and wing shape variation. The principal component analysis and canonical variate analysis revealed morphological variations between the populations studied, which were then confirmed by the permutation test of the differences between populations using Mahalanobis and Procustes distances. The multivariate regression analysis shows that the centroid size influences the shape of the heads and wings. T. infestans of the inter-Andean valleys are longer in the head and wings compared to the population of the Chaco. We propose that the geometric shape variation may be explained by geographical changes in climatic conditions, peri-domiciliar habitats, food source quality, and the use of insecticides.