Genomic differentiation and niche divergence in the Hetaerina americana (Odonata) cryptic species complex.

The evolution of reproductive barriers, that is, the speciation process, implies the limitation of gene flow between populations. Different patterns of genomic differentiation throughout the speciation continuum may provide insights into the causal evolutionary forces of species divergence. In this study, we analysed a cryptic species complex of the genus Hetaerina (Odonata). This complex includes H. americana and H. calverti; however, in H. americana two highly differentiated genetic groups have been previously detected, which, we hypothesize, may correspond to different species with low morphological variation. We obtained single nucleotide polymorphism (SNP) data for 90 individuals belonging to the different taxa in the complex and carried out differentiation tests to identify genetic isolation. The results from STRUCTURE and discriminant analysis of principal components (DAPC), based on almost 5000 SNPs, confirmed the presence of three highly differentiated taxa. Also, we found FST values above 0.5 in pairwise comparisons, which indicates a considerable degree of genetic isolation among the suggested species. We also found low climatic niche overlap among all taxa, suggesting that each group occurs at specific conditions of temperature, precipitation and elevation. We propose that H. americana comprises two cryptic species, which may be reproductively isolated by ecological barriers related to niche divergence, since the morphological variation is minimal and, therefore, mechanical barriers are probably less effective compared to other related species such as H. calverti.

A chromosome-level genome assembly for the smoky rubyspot damselfly (Hetaerina titia).

Smoky rubyspot damselflies (Hetaerina titia Drury, 1773) are one of the most commonly encountered odonates along streams and rivers on both slopes of Central America and the Atlantic drainages in the United States and southern Canada. Owing to their highly variable wing pigmentation, they have become a model system for studying sexual selection and interspecific behavioral interference. Here, we sequence and assemble the genome of a female smoky rubyspot. Of the primary assembly (i.e. the principle pseudohaplotype), 98.8% is made up of 12 chromosomal pseudomolecules (2N = 22A + X). There are 75 scaffolds in total, an N50 of 120 Mb, a contig-N50 of 0.64 Mb, and a high arthropod BUSCO score [C: 97.6% (S: 97.3%, D: 0.3%), F: 0.8%, M: 1.6%]. We then compare our assembly to that of the blue-tailed damselfly genome (Ischnura elegans), the most complete damselfly assembly to date, and a recently published assembly for an American rubyspot damselfly (Hetaerina americana). Collectively, these resources make Hetaerina a genome-enabled genus for further studies of the ecological and evolutionary forces shaping biological diversity.

Complex evolutionary history of the American Rubyspot damselfly, Hetaerina americana (Odonata): Evidence of cryptic speciation.

Analyzing the magnitude and distribution of genetic variation within and among populations allows for hypothesis testing about historical demographic size changes, secondary contacts, refugia, and speciation patterns. Species distribution and genetic structure are greatly influenced by the complex life cycle and behavior of odonates. Hetaerina americana has been widely used as a model system in behavioral studies, but its population genetic structure has not been analyzed, except for a single study that included only three populations but identified the presence of markedly differentiated genetic groups, suggesting the existence of cryptic species. Here, we tested this hypothesis by assessing throughout the distribution range of H. americana the patterns of genetic and morphological variation in the male caudal appendages, due to the great importance of these structures in mate recognition. As molecular markers we used sequences of the mitochondrial cytochrome oxidase I (COI) gene and the nuclear internal transcribed spacer (ITS) region, as well as six nuclear microsatellites. We found very high population genetic differentiation (ΦST > 0.51) in the three sets of markers but with strong mitonuclear discordance. A neutrality test suggested that the mitochondrial genome might be under purifying selection in association to climatic variables (temperature seasonality). The assignment of individuals to nuclear genetic groups showed little admixture and complete congruence with morphological differentiation in the male caudal appendages. Hence, the results suggest that H. americana represents at least two different cryptic species which are isolated reproductively.

The costs of the immune memory within generations.

Immune response is evolutionary costly, but it is not clear whether these costs affect energetic expenditure (short-term cost), growth (medium-term cost), or reproduction (long-term cost). We tested the costs of immune memory in Tenebrio molitor against Metarhizium brunneum. To do this, we used two groups of T. molitor larvae: (a) the control group, which was injected first with Tween solution and 10 days later with M. brunneum and (b) the memory group, which was first injected with M. brunneum and 10 days later with M. brunneum. Compared to controls, larvae of the memory group were more likely to survive, but they also had an increased metabolic rate (CO2 production), spent a long time before becoming pupae, and had a shorter time from pupae to adulthood. In the adult stage, control females preferred control males, but there was no significant difference in the preference of memory females. Finally, control and memory males preferred control females. These results confirm that immune memory has costs in terms of energetic expenditure, growth, and reproduction. To the best of our knowledge, this is the first experimental demonstration that immune memory in larvae is traded-off with adult sexual selection involving mate choice.

The effect of colour polymorphism on thermoregulation in an orb web spider.

Spiders that build aerial webs in open areas face the risk of overheating due to incident solar radiation. These spiders can counteract overheating by either moving the web to another site or by adopting behavioural thermoregulation within the web. Since moving can be costly, studies have suggested that a passive but effective method of reducing heat load is by light reflectance through body colouration. We explored the interaction between colour and thermoregulation in a colour polymorphic species, under both field and laboratory conditions. We show that in natural conditions, the spiders maintain their body temperature below that of the ambient, but with no difference in surface temperature between colour morphs. In laboratory experiments with internal temperature measurements, white morphs bore the risk of overheating better than the yellow morphs since they heated up slower and cooled faster. We suggest that the thermoregulatory properties of colour polymorphism in Verrucosa arenata have physiological consequences and may play an important role in the maintenance of colour polymorphism in this species.

Reproductive benefits and reduced investment in parental care behavior associated with reproductive groups of males in Abudefduf troschelii.

Fishes of the family Pomacentridae present a wide diversity of mating systems, ranging from polygyny to promiscuity and from individual territorial defense to the establishment of reproductive colonies of males. The damselfish species Abudefduf troschelii has a reproductive colony mating system, in which males temporarily aggregate in reproductive areas to court and attract females. Males defend an individual territory where they receive eggs and perform paternal care behaviors for their offspring. The present study evaluated the advantages of the colonial mating system in A. troschelii. During an entire reproductive period, in a breeding colony within a rocky reef, we located, marked, geo-referenced, and measured the distances between the territories of all males. We quantified the variance among males in their patterns of paternal care investment, eggs acquired, hatching success, reproductive success, body size, and changes in body coloration. We found that males spatially distributed their nests in groups or independently (i.e., solitary nests). Nesting groups are formed by larger males that show intense nuptial coloration during the entire receptivity period. They are located centrally to the colony and consist of three to six males whose territories overlap. In contrast, small solitary males that fail to acquire or maintain nuptial coloration during the receptivity period establish their nests peripherally to the colony, away from the territories of other males. Our results highlight that the reproductive benefits of colonial nesting are unequal for males, as the spatial distribution of nests within the colony determines the reproductive success of males. Group nesting confers the highest reproductive benefits to males regarding eggs obtained, hatching success, and relative fitness and also enables males to reduce their parental investment in brood care behaviors. The preference of females for oviposition could be associated with greater intrasexual competitiveness, defense ability, body condition, or experience of group-nesting males located at the center of the colony or because their progeny will have a lower probability of predation than they would in solitary nests males.

Living on the edge: urban fireflies (Coleoptera, Lampyridae) in Morelia, Michoacán, Mexico.

Fireflies (Coleoptera, Lampyridae) are a globally threatened group of insects due to habitat loss and fragmentation, light pollution, climate change and pesticides. However, against all odds, some firefly populations persist in urbanized environments where all four of these factors are present simultaneously. In this work, we compiled several data sources to document the diversity of fireflies in the urbanized area of Morelia, characterize their current habitats, and determine the main stressors affecting these bioluminescent insects. We found seven genera and 26 species of fireflies (19 nocturnal, seven diurnal) associated with 32 urban, peri-urban and extra-urban areas; at least, 14 are new records for Michoacán, and the list for the state now includes nine genera and 41 species. Five additional sites were documented as extinction sites. We compared the characteristics of these five sites with those of the sites with extant populations. We found that in Morelia, fireflies are mainly associated with areas that have high to moderate proportions of vegetation cover, are near water bodies, have very gentle to moderate slopes, and are exposed to low levels of light pollution. In contrast, the extinction sites showed high proportions of artificial surfaces and high levels of light pollution. Because some fireflies are considered bioindicators of ecosystem integrity as they are associated to specific habitats, are highly diverse and due to their sensitivity to environmental changes, we consider that sites from Morelia's urban core and extinction sites show the highest levels of environmental degradation, threatening most fireflies and other insects living in the urban core with local extinction. At the same time, our results also suggest that implementing conservation strategies and sustainable planning for the urban development of Morelia in the short term could allow fireflies and other vital elements of the city's insect communities to persist for future generations. Restoration and conservation of green areas and nighttime environments are essential for biodiversity and human health, especially in intra-urban zones.

Color and morphological differentiation in the Sinaloa Wren (Thryophilus sinaloa) in the tropical dry forests of Mexico: The role of environment and geographic isolation.

The role and the degree to which environment and geographic isolation contribute to phenotypic diversity has been widely debated. Here, we studied phenotypic variation (morphology and plumage reflectance) in the Sinaloa Wren, an endemic bird distributed throughout the tropical dry forest (TDF) on the Mexican pacific slope where a pronounced variability in environmental conditions has been reported. In particular, we aimed: 1) to characterize phenotypic variation between subspecies; 2) to analyze the relationship between phenotypic and environmental variation in the context of classic ecogeographic rules, such as Bergmann's, Allen's, Gloger's, and Bogert's, and to quantify the relative roles of environment and geographic isolation and their interaction in shaping phenotypic variation; and 3) to test for niche conservatism between subspecies. Our data revealed significant differences among subspecies morphology and plumage reflectance. The environment explained a higher proportion of the morphological variation, while geography explained a smaller proportion. However, variation in plumage reflectance was mainly explained by the joint effect of geography and environment. Our data did not support for Bergmann´s and Allen´s rule. However, longer tails and wings were positively associated with higher elevations, larger tarsus and culmens were positively related to higher latitudes and to greater tree cover, respectively. Our data partially supported Gloger´s rule, where darker plumages were associated with more humid environments. The effects of temperature on plumage coloration were more consistent with Bogert´s rule. In addition, we found darker plumages related to higher levels of UV-B radiation. Finally, niche divergence was detected between T. s. cinereus and T. s. sinaloa vs. T. s. russeus. In a continuously distributed ecosystem such as the TDF on the pacific slope of Mexico, the environmental conditions and geographic isolation have played an important role in promoting phenotypic differentiation in the Sinaloa Wren.

Morphological variation and reproductive isolation in the Hetaerina americana species complex.

Incomplete premating barriers in closely related species may result in reproductive interference. This process has different fitness consequences and can lead to three scenarios: niche segregation, sexual exclusion, or reproductive character displacement. In morphologically cryptic species, isolation barriers can be difficult to recognize. Here, we analyzed the morphological, behavioral, and genetic differences between two sympatric cryptic species of the genus Hetaerina to determine the characters that contribute the most to reproductive isolation and the effect of the high rates of behavior interference between the species. We found complete genetic isolation and significant differences in the morphometry of caudal appendages and wing shape, as well as body size variation between species. In contrast, we did not find clear differences in the coloration of the wing spot and observed high rates of interspecific aggression. Our results suggest that divergence in the shape of the caudal appendages is the principal pre-mating barrier that prevents interspecific mating. Moreover, a scenario of character displacement on body size was found. Nevertheless, size could play an important role in both inter- and intrasexual interactions and, therefore, we cannot differentiate if it has resulted from reproductive or aggressive interference.

The use of highly diverse clades as a surrogate for habitat integrity analysis: Argia damselflies as a practical tool for rapid assessments.

Human activities have impacted many environments on earth, and thus several species are facing an increased risk of extinction. The environmental crisis requires rapid tools to assess the ecosystem health accurately. Studies have been conducted with visual indices that quantify habitat integrity by predicting species richness and diversity. However, whether a diverse clade can predict habitat integrity has not been used. The genus Argia (Rambur, 1842) is one of the most locally diverse groups in southeastern Mexico. In this context, we hypothesized that the occurrence, species richness, and diversity of adults Argia spp. could be a better predictor of the Visual-Based Habitat Assessment Score (VBHAS) than the other taxonomic levels or less diverse clades. We found that the richness and diversity of Argia spp. are positively correlated with VBHA scores, as same as taxonomic ratios. Simultaneously, VBHA scores increase to 23.51 times when Argia spp. diversity increases. We discuss the possible use of a diverse Odonata clade, as Argia spp. could surrogate habitat integrity for local long-term biomonitoring programs. This approach requires testing with other indices and verifying a reliable and consistent relationship between diverse clades and environmental assessment scores.

The use of Odonata species for environmental assessment: a meta-analysis for the Neotropical region.

The order Odonata has been regularly used as an indicator of the ecosystem's condition. The objective of this review was to analyze the importance of Odonata for environmental assessments (assessment types, statistical approach, life stages, and sampling method, or particular metric), summarizing the current state, the trends, and identifying related research issues in the Neotropical region. Therefore, we selected 62 articles from 2007 to 2018 based on published research to monitor Odonata assessments in the Neotropical region. We compiled a database and ran statistical analyses for the observed frequencies. We found that ecosystem health was the most frequent assessment type and quality the most used objective. In the case of statistical tests and metrics, multivariate analyses and species richness were most used in these papers. However, because there is a great diversity of habitats in this region, there is no unique monitoring protocol to assess the quality of ecosystem health and it is needed to create a proposal for a standard evaluation protocol. Consequently, guidelines for monitoring are presented, and we suggest three stages to establish a specific protocol for each site, which records the set of species most sensitive to the exchange rate evaluated, as well as the use of rarefaction methods, the index of diversity based on the area under the curve, and multivariate analysis, among other recommendations.

Interspecific gene flow in a multispecies oak hybrid zone in the Sierra Tarahumara of Mexico.

BACKGROUND AND AIMS: Interspecific gene flow can occur in many combinations among species within the genus Quercus, but simultaneous hybridization among more than two species has been rarely analysed. The present study addresses the genetic structure and morphological variation in a triple hybrid zone formed by Q. hypoleucoides, Q. scytophylla and Q. sideroxyla in north-western Mexico. METHODS: A total of 247 trees from ten reference and 13 presumed intermediate populations were characterized using leaf shape variation and geometric morphometrics, and seven nuclear microsatellites as genetic markers. Discriminant function analysis was performed for leaf shape variation, and estimates of genetic diversity and structure, and individual Bayesian genetic assignments were obtained. KEY RESULTS: Reference populations formed three completely distinct groups according to discriminant function analysis based on the morphological data, and showed low, but significant, genetic differentiation. Populations from the zone of contact contained individuals morphologically intermediate between pairs of species in different combinations, or even among the three species. The Bayesian admixture analysis found that three main genetic clusters best fitted the data, with good correspondence of reference populations of each species to one of the genetic clusters, but various degrees of admixture evidenced in populations from the contact area. CONCLUSIONS: The three oak species have formed a complex hybrid zone that is geographically structured as a mosaic, and comprising a wide range of genotypes, including hybrids between different species pairs, backcrosses and probable triple hybrids.

Hetaerina calverti (Odonata: Zygoptera: Calopterygidae) sp. nov., a new cryptic species of the American Rubyspot complex.

Hetaerina americana Fabricius, 1798 has a long and irresolute taxonomic history. Several synonyms have been suggested (H. californica Hagen in Selys-Longchamps, 1859, H. basalis Hagen in Selys-Longchamps, 1859, H. texana Walsh, 1863, H. scelerata Walsh, 1863, etc.), related to the variation in the size of the wing spots as well as to the morphology of the male cerci. However, Calvert (1901) suggested that H. americana represents one variable species. Nevertheless, Vega-Sánchez et al. (2019) through a genetic and morphological analysis presented evidence to propose that H. americana represents a species complex. In the present work, we describe a new species that belongs to this complex: H. calverti sp. nov. The morphological characteristics by which males and females of H. calverti differ from H. americana are highlighted. The most important character for the differentiation of males is the shape of the cerci and the size of the individuals (when the two species are in sympatry). In females, the main differences are in the shape of the intersternites and the medio-dorsal carina of the last segment of the abdomen. Some generalities about the biology of the species are presented, including geographical distribution patterns and genetic divergence data. [urn:lsid:zoobank.org:act:F5C329AE-7A00-4979-8A0D-A13D869E54B1].

Sex-related interannual plasticity in wing morphological design in Heliconius charithonia enhances flight metabolic performance.

Flight morphological variations and its consequences on animal performance are common in winged insects. In the butterfly Heliconius charithonia, sex-related differences in the wing morphological design have been described resulting in differences in foraging behavior, daily flight distances and flight aerodynamics. It has been suggested that these differences should be reflected in the metabolic capacities and energetic budgets associated with flight in both sexes. In this study, we analyzed the relationship between wing morphological variation and metabolic performance, flight aerodynamics and energetic reserves in females and males of Heliconius charithonia over two years. The results confirm the presence of wing shape sexual dimorphism, but also show an unexpected sex-related annual variation in wing shape, mirrored in the metabolic condition (resting metabolic rate) of individuals. However, contrary to expectation, intersexual variations in wing shape are not related to differences between the sexes in terms of flight aerodynamics, flight metabolic rates, or energetic reserves (carbohydrates, lipids and proteins). Our results indicate a considerable plasticity in H. charithonia wing shape, which we suggest is determined by a trade-off between environmental pressures and reproductive restriction of each sex, maintaining an optimum flight design. Finally, similarities in metabolic rates between young and older males and females in both years may be a consequence of the ability of Heliconius species to feed on pollen.

Reduced thermal variability in cities and its impact on honey bee thermal tolerance.

Urbanization is one of the most significant land cover transformations, and while climate alteration is one of its most cited ecological consequences we have very limited knowledge on its effect on species' thermal responses. We investigated whether changes in environmental thermal variability caused by urbanization influence thermal tolerance in honey bees (Apis mellifera) in a semi-arid city in central Mexico. Ambient environmental temperature and honey bee thermal tolerance were compared in urban and rural sites. Ambient temperature variability decreased with urbanization due to significantly higher nighttime temperatures in urban compared to rural sites and not from differences in maximum daily temperatures. Honey bee thermal tolerance breadth [critical thermal maxima (CTmax)-critical thermal minima (CTmin)] was narrower for urban bees as a result of differences in cold tolerance, with urban individuals having significantly higher CTmin than rural individuals, and CTmax not differing among urban and rural individuals. Honey bee body size was not correlated to thermal tolerance, and body size did not differ between urban and rural individuals. We found that honey bees' cold tolerance is modified through acclimation. Our results show that differences in thermal variability along small spatial scales such as urban-rural gradients can influence species' thermal tolerance breadths.

Do territorial butterflies show a macroecological fighting pattern in response to environmental stability?

The territorial defense of mating sites by males should be favored when female monopolization is possible. Such monopolization should occur in species in which females emerge asynchronously, since males may have time to copulate with one female before the arrival of other. However, regions with smaller reproductive windows should promote higher synchronicity of female emergence, generating a predictable macroecological pattern associated to the rewards from territorial defense. In this study we evaluated the hypothesis that territorial male butterflies should invest more in fighting in species that occur in areas with stable climatic conditions that should present longer reproductive windows. We compiled studies reporting mean butterfly fighting times, mean trait differences among winners and losers and local Köppen climatic classification (a surrogate for climatic stability). We found that males from butterfly species located in areas with stable climatic conditions fight for longer times. However, although winners were stronger than intruders only in areas with intermediate climatic conditions, there was a marked variation among winner-loser comparisons in species in areas with stable climatic conditions. We conclude that males from butterfly species that occur in areas with stable climatic regimes invest more in fighting due to the higher payoffs accrued with territorial defense, but that such investment does not change the effect of trait asymmetries in settling territorial conflicts. This article is part of a Special Issue entitled: Special Issue:Neotropical Behaviour.