Independent transitions to freshwater environments promote phenotypic divergence, not convergence, in stingrays.

Instances of convergent or parallel evolution provide a potent model system for exploring contingency and determinism in evolutionary biology. Likewise, the multiple, independent habitat transitions from saltwater to freshwater biomes offer opportunity for studying convergent evolution within and among different vertebrate lineages. For example, stingrays have invaded freshwater habitats multiple times across different continents, sometimes even several times within the same clade (e.g., Dasyatidae). We evaluated the frequency of saltwater-freshwater invasions in stingrays, compared ecological and phenotypic diversification among freshwater and saltwater lineages, and assessed the degree of convergence among freshwater species. Despite not being morphologically distinct from saltwater stingrays, freshwater stingrays do expand the margins of stingray morphological diversity. According to our data, trophic specialists occupied non-overlapping regions of morphospace, with piscivores and molluscivores being distinct from other diet guilds. Freshwater stingrays as a group did not strongly converge morphologically, neither did freshwater rays from different lineages which shared similar niches. These findings could be explained by there not being enough time for convergence to occur among more ancient and more recent freshwater lineages. Alternatively, the different ancestral bauplans of various freshwater ray lineages and weak selection on optimal phenotypes could promote contingency in the form of evolution along paths of least resistance.

True grit? Comparative anatomy and evolution of gizzards in fishes.

Gut morphology frequently reflects the food organisms digest. Gizzards are organs of the gut found in archosaurs and fishes that mechanically reduce food to aid digestion. Gizzards are thought to compensate for edentulism and/or provide an advantage when consuming small, tough food items (e.g., phytoplankton and algae). It is unknown how widespread gizzards are in fishes and how similar these structures are among different lineages. Here, we investigate the distribution of gizzards across bony fishes to (1) survey different fishes for gizzard presence, (2) compare the histological structure of gizzards in three species, (3) estimate how often gizzards have evolved in fishes, and (4) explore whether anatomical and ecological traits like edentulism and microphagy predict gizzard presence. According to our analyses, gizzards are rare across bony fishes, evolving only six times in a broad taxonomic sampling of 51 species, and gizzard presence is not clearly correlated with factors like gut length or dentition. We find that gizzard morphology varies among the lineages where one is present, both macroscopically (presence of a crop) and microscopically (varying tissue types). We conclude that gizzards likely aid in the mechanical reduction of food in fishes that have lost an oral dentition in their evolutionary past; however, the relative scarcity of gizzards suggests they are just one of many possible solutions for processing tough, nutrient-poor food items. Gizzards have long been present in the evolutionary history of fishes, can be found in a wide variety of marine and freshwater clades, and likely have been overlooked in many taxa.

How to Survive a (Juvenile) Piranha Attack: An Integrative Approach to Evaluating Predator Performance.

Figures: Cory cat panel figureDrawing of bite force measuring equipment and indentation rig Pygocentrus nattereri jaw muscle morphology and skull anatomyBox plot grid of number of Pygocentrus nattereri bites before puncture along different body regions of Corydoras trilineatus during feeding trials resultsDrawing of color-coded Corydoras trilineatus with attack frequencies and average bites until puncture by Pygocentrus nattereriBox plot of average voluntary juvenile Pygocentrus nattereri bite forces to standard lengthPanel of linear ordinary least-squares regressions of Pygocentrus nattereri bite force to adductor mandibulae mass, standard length, and body massOrdinary least-squares regressions of voluntary bites to restrained bites of Pygocentrus nattereriPanel of indentation tests for intact and removed Corydoras trilineatus scutesPanel of indentation tests for Corydoras trilineatus body region. Synopsis: There is an evolutionary arms race between predators and prey. In aquatic environments, predatory fishes often use sharp teeth, powerful bites, and/or streamlined bodies to help capture their prey quickly and efficiently. Conversely, prey are often equipped with antipredator adaptations including: scaly armor, sharp spines, and/or toxic secretions. This study focused on the predator-prey interactions between the armored threestripe cory catfish (Corydoras trilineatus) and juvenile red-bellied piranha (Pygocentrus nattereri). Specifically, we investigated how resistant cory catfish armor is to a range of natural and theoretical piranha bite forces and how often this protection translated to survival from predator attacks by Corydoras. We measured the bite force and jaw functional morphology of P. nattereri, the puncture resistance of defensive scutes in C. trilineatus, and the in situ predatory interactions between the two. The adductor mandibulae muscle in juvenile P. nattereri is robust and delivers an average bite force of 1.03 N and maximum bite force of 9.71 N, yet its prey, C. trilineatus, survived 37% of confirmed bites without any damage. The C. trilineatus armor withstood an average of nine bites before puncture by P. nattereri. Predation was successful only when piranhas bit unarmored areas of the body, at the opercular opening and at the caudal peduncle. This study used an integrative approach to understand the outcomes of predator-prey interactions by evaluating the link between morphology and feeding behavior. We found that juvenile P. nattereri rarely used a maximal bite force and displayed a net predation success rate on par with other adult vertebrates. Conversely, C. trilineatus successfully avoided predation by orienting predator attacks toward their resilient, axial armor and behavioral strategies that reduced the predator's ability to bite in less armored regions of the body.

The Lesser-Known Transitions: Organismal Form and Function across Abiotic Gradients.

From minute-to-minute changes, or across daily, seasonal, or geological timescales, animals are forced to navigate dynamic surroundings. Their abiotic environment is continually changing. These changes could include alterations to the substrates animals locomote on, flow dynamics of the microhabitats they feed in, or even altitudinal shifts over migration routes. The only constancy in any organism's day-to-day existence is the heterogeneity of the habitats they move through and the gradients in the physical media (e.g., air and water) they live in. We explored a broad range of organismal transitions across abiotic gradients and investigated how these organisms modify their form, function, and behavior to accommodate their surrounding media. We asked the following questions: (1) What are some challenges common to animals in changing media or moving between media? (2) What are common solutions to these recurring problems? (3) How often are these common solutions instances of either convergence or parallelism? Our symposium speakers explored these questions through critical analysis of numerous datasets spanning multiple taxa, timescales, and levels of analysis. After discussions with our speakers, we suggest that the role of physical principles (e.g., drag, gravity, buoyancy, and viscosity) in constraining morphology and shaping the realized niche has been underappreciated. We recommend that investigations of these transitions and corresponding adaptations should include comparisons at multiple levels of biological organization and timescale. Relatedly, studies of organisms that undergo habitat and substrate changes over ontogeny would be worthwhile to include in comparisons. Future researchers should ideally complement lab-based morphological and kinematic studies with observational and experimental approaches in the field. Synthesis of the findings of our speakers across multiple study systems, timescales, and transitional habitats suggests that behavioral modification and exaptation of morphology play key roles in modulating novel transitions between substrates.

Ecological and Phenotypic Diversification after a Continental Invasion in Neotropical Freshwater Stingrays.

Habitat transitions are key potential explanations for why some lineages have diversified and others have not-from Anolis lizards to Darwin's finches. The ecological ramifications of marine-to-freshwater transitions for fishes suggest evolutionary contingency: some lineages maintain their ancestral niches in novel habitats (niche conservatism), whereas others alter their ecological role. However, few studies have considered phenotypic, ecological, and lineage diversification concurrently to explore this issue. Here, we investigated the macroevolutionary history of the taxonomically and ecologically diverse Neotropical freshwater river rays (subfamily Potamotrygoninae), which invaded and diversified in the Amazon and other South American rivers during the late Oligocene to early Miocene. We generated a time-calibrated, multi-gene phylogeny for Potamotrygoninae and reconstructed evolutionary patterns of diet specialization. We measured functional morphological traits relevant for feeding and used comparative phylogenetic methods to examine how feeding morphology diversified over time. Potamotrygonine trophic and phenotypic diversity are evenly partitioned (non-overlapping) among internal clades for most of their history, until 20-16 mya, when more recent diversification suggests increasing overlap among phenotypes. Specialized piscivores (Heliotrygon and Paratrygon) evolved early in the history of freshwater stingrays, while later trophic specialization (molluscivory, insectivory, and crustacivory) evolved in the genus Potamotrygon. Potamotrygonins demonstrate ecological niche lability in diets and feeding apparatus; however, diversification has mostly been a gradual process through time. We suggest that competition is unlikely to have limited the potamotrygonine invasion and diversification in South America.

Hyperspectral data as a biodiversity screening tool can differentiate among diverse Neotropical fishes.

Hyperspectral data encode information from electromagnetic radiation (i.e., color) of any object in the form of a spectral signature; these data can then be used to distinguish among materials or even map whole landscapes. Although hyperspectral data have been mostly used to study landscape ecology, floral diversity and many other applications in the natural sciences, we propose that spectral signatures can be used for rapid assessment of faunal biodiversity, akin to DNA barcoding and metabarcoding. We demonstrate that spectral signatures of individual, live fish specimens can accurately capture species and clade-level differences in fish coloration, specifically among piranhas and pacus (Family Serrasalmidae), fishes with a long history of taxonomic confusion. We analyzed 47 serrasalmid species and could distinguish spectra among different species and clades, with the method sensitive enough to document changes in fish coloration over ontogeny. Herbivorous pacu spectra were more like one another than they were to piranhas; however, our method also documented interspecific variation in pacus that corresponds to cryptic lineages. While spectra do not serve as an alternative to the collection of curated specimens, hyperspectral data of fishes in the field should help clarify which specimens might be unique or undescribed, complementing existing molecular and morphological techniques.

Phylogenomics of Piranhas and Pacus (Serrasalmidae) Uncovers How Dietary Convergence and Parallelism Obfuscate Traditional Morphological Taxonomy.

The Amazon and neighboring South American river basins harbor the world's most diverse assemblages of freshwater fishes. One of the most prominent South American fish families is the Serrasalmidae (pacus and piranhas), found in nearly every continental basin. Serrasalmids are keystone ecological taxa, being some of the top riverine predators as well as the primary seed dispersers in the flooded forest. Despite their widespread occurrence and notable ecologies, serrasalmid evolutionary history and systematics are controversial. For example, the sister taxon to serrasalmids is contentious, the relationships of major clades within the family are inconsistent across different methodologies, and half of the extant serrasalmid genera are suggested to be non-monophyletic. We analyzed exon capture to reexamine the evolutionary relationships among 63 (of 99) species across all 16 serrasalmid genera and their nearest outgroups, including multiple individuals per species to account for cryptic lineages. To reconstruct the timeline of serrasalmid diversification, we time-calibrated this phylogeny using two different fossil-calibration schemes to account for uncertainty in taxonomy with respect to fossil teeth. Finally, we analyzed diet evolution across the family and comment on associated changes in dentition, highlighting the ecomorphological diversity within serrasalmids. We document widespread non-monophyly of genera within Myleinae, as well as between Serrasalmus and Pristobrycon, and propose that reliance on traits like teeth to distinguish among genera is confounded by ecological homoplasy, especially among herbivorous and omnivorous taxa. We clarify the relationships among all serrasalmid genera, propose new subfamily affiliations, and support hemiodontids as the sister taxon to Serrasalmidae. [Characiformes; exon capture; ichthyochory; molecular time-calibration; piscivory.].

Swimming and defence: competing needs across ontogeny in armoured fishes (Agonidae).

Biological armours are potent model systems for understanding the complex series of competing demands on protective exoskeletons; after all, armoured organisms are the product of millions of years of refined engineering under the harshest conditions. Fishes are no strangers to armour, with various types of armour plating common to the 400-500 Myr of evolution in both jawed and jawless fishes. Here, we focus on the poachers (Agonidae), a family of armoured fishes native to temperate waters of the Pacific rim. We examined armour morphology, body stiffness and swimming performance in the northern spearnose poacher (Agonopsis vulsa) over ontogeny. As juveniles, these fishes make frequent nocturnal forays into the water column in search of food, while heavily armoured adults are bound to the benthos. Most armour dimensions and density increase with body length, as does body stiffness. Juvenile poachers have enlarged spines on their armour whereas adults invest more mineral in armour plate bases. Adults are stiffer and accelerate faster than juveniles with an anguilliform swimming mode. Subadults more closely approximate adults more than smaller juveniles, with regards to both swimming and armour mechanics. Poacher armour serves multiple functions over ontogeny, from facilitating locomotion, slowing sinking and providing defence.

The Natural Historian's Guide to the CT Galaxy: Step-by-Step Instructions for Preparing and Analyzing Computed Tomographic (CT) Data Using Cross-Platform, Open Access Software.

The decreasing cost of acquiring computed tomographic (CT) data has fueled a global effort to digitize the anatomy of museum specimens. This effort has produced a wealth of open access digital three-dimensional (3D) models of anatomy available to anyone with access to the Internet. The potential applications of these data are broad, ranging from 3D printing for purely educational purposes to the development of highly advanced biomechanical models of anatomical structures. However, while virtually anyone can access these digital data, relatively few have the training to easily derive a desirable product (e.g., a 3D visualization of an anatomical structure) from them. Here, we present a workflow based on free, open source, cross-platform software for processing CT data. We provide step-by-step instructions that start with acquiring CT data from a new reconstruction or an open access repository, and progress through visualizing, measuring, landmarking, and constructing digital 3D models of anatomical structures. We also include instructions for digital dissection, data reduction, and exporting data for use in downstream applications such as 3D printing. Finally, we provide Supplementary Videos and workflows that demonstrate how the workflow facilitates five specific applications: measuring functional traits associated with feeding, digitally isolating anatomical structures, isolating regions of interest using semi-automated segmentation, collecting data with simple visual tools, and reducing file size and converting file type of a 3D model.

PORTUGUÊS (PORTUGUESE)  O Guia da Galáxia da Tomografia Computadorizada para um Biólogo: instruções passo a passo para preparar e analisar dados tomográficos usando um software gratuito de acesso aberto  Thaddaeus J. Buser, Olivia F. Boyd, Álvaro Cortés, Cassandra M. Donatelli, Matthew A. Kolmann, Jennifer L. Luparell, Janne A. Pfeiffenberger, Brian L. Sidlauskas, Adam P. Summers  RESUMOO custo decrescente da obtenção de dados de Tomografia Computadorizada (TC) alimentou um esforço global para digitalizar espécimes depositados em museus. Esse esforço produziu uma grande variedade de modelos digitais 3 D com dados de anatomia, disponíveis para qualquer pessoa com acesso à Internet. As aplicações potenciais desses dados são amplas, desde a impressão 3 D para fins puramente educacionais, até o desenvolvimento de modelos biomecânicos de estruturas anatômicas altamente avançados. No entanto, enquanto praticamente qualquer pessoa pode acessar esses dados digitais, relativamente poucos têm o treinamento para obter facilmente um produto de interesse (por exemplo, uma visualização 3 D de uma estrutura anatômica). Aqui, apresentamos um tutorial baseado em um software gratuito de código aberto e multiplataforma para o processamento de dados de TC. Fornecemos instruções passo a passo que começam com a obtenção de dados de TC a partir de uma nova reconstrução ou num repositório de acesso aberto, e progredimos através da visualização, medição, marca de referência e construção de modelos digitais 3 D de estruturas anatômicas. Também incluímos instruções para dissecação digital, redução de dados e exportação de dados para uso em aplicativos posteriores, como os de impressoras 3 D. Por fim, fornecemos vídeos e tutoriais suplementares que demonstram como o tutorial facilita cinco aplicações específicas: medir características funcionais associadas à alimentação, isolar estruturas anatômicas digitalmente, isolar regiões de interesse usando segmentação semi-automática, coletar dados com ferramentas visuais simples, e reduzir o tamanho de arquivo e converter o tipo de arquivo do modelo 3 D.

FRANÇAIS (FRENCH)  Guide de l'historien de la nature à travers la galaxie TDM: instructions étape par étape pour la préparation et l'analyse de données tomodensitométrique (TDM) à l'aide d'un logiciel à accès ouvert multiplateforme Thaddaeus J. Buser, Olivia F. Boyd, Álvaro Cortés, Cassandra M. Donatelli, Matthew A. Kolmann, Jennifer L. Luparell, Janne A. Pfeiffenberger, Brian L. Sidlauskas, Adam P. Summers RÉSUMÉLe coût décroissant de l'acquisition de données tomodensitométriques (TDM) a alimenté un effort mondial pour numériser l'anatomie des spécimens de musée. Cet effort a produit une multitude de modèles d'anatomie numérique 3 D en accès libre accessibles à tous ceux qui ont accès à Internet. Les applications potentielles de ces données sont vastes, allant de l'impression 3 D à des fins purement pédagogiques au développement de modèles biomécaniques de structures anatomiques très avancés. Cependant, alors que pratiquement tout le monde peut accéder à ces données numériques, relativement peu ont la formation nécessaire pour en tirer facilement un produit intéressant (par exemple, une visualisation 3 D d'une structure anatomique). Ici, nous présentons un flux de travail basé sur un logiciel gratuit, à accès ouvert et multiplateforme pour le traitement des données TDM. Nous fournissons des instructions étape par étape qui commencent par l'acquisition de données TDM à partir d'une nouvelle reconstruction ou d'un référentiel en accès gratuit, et progressent à travers la visualisation, la mesure, le marquage et la construction de modèles numériques 3 D de structures anatomiques. Nous incluons également des instructions pour la dissection numérique, la réduction des données et l'exportation de données à utiliser dans des applications en aval telles que l'impression 3 D. Enfin, nous proposons des vidéos et des workflows supplémentaires qui montrent comment le workflow facilite cinq applications spécifiques: mesurer les traits fonctionnels associés à l'alimentation, isoler numériquement les structures anatomiques, isoler les régions d'intérêt à l'aide de la segmentation semi-automatisée, collecter des données avec des outils visuels simples, réduire la taille du fichier et convertir le type de fichierd'un modèle 3 D.

Have Niche, Will Travel. New Means of Linking Diet and Ecomorphology Reveals Niche Conservatism in Freshwater Cottoid Fishes.

Evolutionary transitions between habitats have been catalysts for some of the most stunning examples of adaptive diversification, with novel niches and new resources providing ecological opportunity for such radiations. In aquatic animals, transitions from saltwater to freshwater habitats are rare, but occur often enough that in the Neotropics for example, marine-derived fishes contribute noticeably to regional ichthyofaunal diversity. Here, we investigate how morphology has evolved in a group of temperate fishes that contain a marine to freshwater transition: the sculpins (Percomorpha; Cottoidea). We devised a novel method for classifying dietary niche and relating functional aspects of prey to their predators. Coupled with functional measurements of the jaw apparatus in cottoids, we explored whether freshwater sculpins have fundamentally changed their niche after invading freshwater (niche lability) or if they retain a niche similar to their marine cousins (niche conservatism). Freshwater sculpins exhibit both phylogeographical and ecological signals of phylogenetic niche conservatism, meaning that regardless of habitat, sculpins fill similar niche roles in either saltwater or freshwater. Rather than competition guiding niche conservatism in freshwater cottoids, we argue that strong intrinsic constraints on morphological and ecological evolution are at play, contra to other studies of diversification in marine-derived freshwater fishes. However, several intertidal and subtidal sculpins as well as several pelagic freshwater species from Lake Baikal show remarkable departures from the typical sculpin bauplan. Our method of prey categorization provides an explicit, quantitative means of classifying dietary niche for macroevolutionary studies, rather than relying on somewhat arbitrary means used in previous literature.

Tem Nicho, Viaja. Novos Meios de Associar Dieta e Ecomorfologia Revelam Conservadorismo de Nicho em Peixes Cotoides de Água Doce (Have Niche, Will Travel. New Means of Linking Diet and Ecomorphology Reveals Niche Conservatism in Freshwater Cottoid Fishes) Transições evolutivas entre habitats têm sido catalisadores de alguns dos mais impressionantes exemplos de diversificação adaptativa, com novos nichos e recursos proporcionando oportunidade ecológica para tais radiações. Em animais aquáticos, as transições de água salgada para habitats de água doce são raras, mas ocorrem com freqüência suficiente para que, nos Neotrópicos, por exemplo, os peixes marinhos contribuam notavelmente para a diversidade regional da ictiofauna. Aqui, nós investigamos como a morfologia evoluiu em um grupo de peixes temperados que contêm uma transição marinha para a água doce: os esculpentes (Percomorpha; Cottoidea). Nós concebemos um novo método para classificar o nicho alimentar e relacionar os aspectos funcionais das presas aos seus predadores. Juntamente com medidas funcionais do aparato de mandíbula em cotoides, exploramos se os esculpentes de água doce mudaram fundamentalmente seu nicho depois de invadi-la (labilidade de nicho) ou se eles mantêm um nicho semelhante aos seus primos marinhos (conservadorismo de nicho). Os esculpentes de água doce exibem sinais filogeográficos e ecológicos de conservadorismo filogenético de nicho, o que significa que, independente do habitat, os esculpentes preenchem papéis ecológicos semelhantes em água salgada ou doce. Mais do que a concorrência guiando o conservadorismo de nicho em cotoides de água doce, argumentamos que fortes restrições intrínsecas à evolução morfológica e ecológica estão em jogo, em contraste com outros estudos de diversificação em peixes de água doce derivados do mar. No entanto, vários esculpentes intertidais e subtidais, bem como várias espécies pelágicas de água doce do Lago Baikal, mostram notáveis desvios do típico bauplan dos esculpentes. Nosso método de categorização de presas fornece um modo explícito e quantitativo de classificar o nicho alimentar para estudos macroevolutivos ao invéz de depender de meios arbitrários usados na literatura anterior. Translated to Portuguese by G. Sobral (gabisobral@gmail.com).