Showing papers in "Zoology in 2017"

The insect spermatheca: an overview.

Abstract: In the female insect, the spermatheca is an ectodermal organ responsible for receiving, maintaining, and releasing sperm to fertilize eggs. The number and morphology of spermathecae vary according to species. Within the spermathecal lumen, substances in the semen and secretions from the spermathecal gland nourish the sperm. Thus, the spermatheca provides an appropriate environment that ensures the long-term viability of sperm. Maintaining sperm viability for long periods within the spermatheca is crucial for insect reproductive success; however, the details of this process remain poorly understood. This review examines several aspects of and gaps in the current understanding of spermatheca biology, including morphology, function, reservoir filling, development, and biochemistry. Despite the importance of the spermatheca in insects, there is little information on the gland secretions and their role in the maintenance and protection of male gametes. Furthermore, in this review, we highlight the current information on spermathecal gland secretions and the likely roles they play in the maintenance and protection of sperm.

Rodent, large animal and non-human primate models of spinal cord injury

Abstract: In this narrative review we aimed to assess the usefulness of the different animal models in identifying injury mechanisms and developing therapies for humans suffering from spinal cord injury (SCI). Results obtained from rodent studies are useful but, due to the anatomical, molecular and functional differences, confirmation of these findings in large animals or non-human primates may lead to basic discoveries that cannot be made in rodent models and that are more useful for developing treatment strategies in humans. SCI in dogs can be considered as intermediate between rodent models and human clinical trials, but the primate models could help to develop appropriate methods that might be more relevant to humans. Ideally, an animal model should meet the requirements of availability and repeatability as well as reproduce the anatomical features and the clinical pathological changing process of SCI. An animal model that completely simulates SCI in humans does not exist. The different experimental models of SCI have advantages and disadvantages for investigating the different aspects of lesion development, recovery mechanisms and potential therapeutic interventions. The potential advantages of non-human primate models include genetic similarities, similar caliber/length of the spinal cord as well as biological and physiological responses to injury which are more similar to humans. Among the potential disadvantages, high operating costs, infrastructural requirements and ethical concerns should be considered. The translation from experimental repair strategies to clinical applications needs to be investigated in future carefully designed studies.

Experimental assessment of the effects of moisture on loggerhead sea turtle hatchling sex ratios.

Abstract: Many reptiles have temperature-dependent sex determination (TSD). Sex determination in marine turtles is described by a cool-male, warm-female pattern. Nest sand temperature strongly influences sea turtle embryo development and sex differentiation. Yet, variation in hatchling sex ratios is explained only partially by nest temperature and can be predicted only at the warmest and coolest temperatures. Hence, other factors during development influence sex determination. Rainfall is a common environmental variable that may impact development and sex determination. We experimentally evaluated bias in sex ratio production associated with nest moisture. Conditions tested in surrogate nests were sand moisture in combination with (i) very restricted evaporation, (ii) moderate evaporation (allowing evaporative cooling), and (iii) evaporative cooling plus cooling from rain-temperature water. We collected eggs from 32 unique loggerhead (Caretta caretta L.) turtle clutches, distributed them among the three different conditions, and incubated the eggs until they hatched. All hatchlings were raised for several months and sex was verified laparoscopically to establish sex ratios for each treatment. The nests were expected to produce 50:50 sex ratios or a moderate female bias (∼70%) based on incubation temperatures. All experimental treatments produced high male bias (87-96%). These results support the hypothesis that moisture impacts sex ratios through evaporation and rainfall-based cooling. High male bias was observed in nests with and without restricted evaporative cooling and no direct cooling due to watering as well as those nests hydrated via cool (rainwater temperature) water. High moisture conditions may produce males through thermal or other mechanisms, highlighting the importance of examining other nest environmental factors on sex determination.

Identification and distribution of neuronal nitric oxide synthase and neurochemical markers in the neuroepithelial cells of the gill and the skin in the giant mudskipper, Periophthalmodon schlosseri

Abstract: Mudskippers are amphibious fishes living in mudflats and mangroves. These fishes hold air in their large buccopharyngeal-opercular cavities where respiratory gas exchange takes place via the gills and higher vascularized epithelium lining the cavities and also the skin epidermis. Although aerial ventilation response to changes in ambient gas concentration has been studied in mudskippers, the localization and distribution of respiratory chemoreceptors, their neurochemical coding and function as well as physiological evidence for the gill or skin as site for O2 and CO2 sensing are currently not known. In the present study we assessed the distribution of serotonin, acetylcholine, catecholamines and nitric oxide in the neuroepithelial cells (NECs) of the mudskipper gill and skin epithelium using immunohistochemistry and confocal microscopy. Colocalization studies showed that 5-HT is coexpressed with nNOS, Na+/K+-ATPase, TH and VAChT; nNOS is coexpressed with Na+/K+-ATPase and TH in the skin. In the gill 5-HT is coexpressed with nNOS and VAhHT and nNOS is coexpressed with Na+/K+-ATPase and TH. Acetylcholine is also expressed in chain and proximal neurons projecting to the efferent filament artery and branchial smooth muscle. The serotonergic cells c labeled with VAChT, nNOS and TH, thus indicating the presence of NEC populations and the possibility that these neurotransmitters (other than serotonin) may act as primary transmitters in the hypoxic reflex in fish gills. Immunolabeling with TH antibodies revealed that NECs in the gill and the skin are innervated by catecholaminergic nerves, thus suggesting that these cells are involved in a central control of branchial functions through their relationships with the sympathetic branchial nervous system. The Na+/K+-ATPase in mitochondria-rich cells (MRCs), which are most concentrated in the gill lamellar epithelium, is colabeled with nNOS and associated with TH nerve terminals. TH-immunopositive fine varicosities were also associated with the numerous capillaries in the skin surface and the layers of the swollen cells. Based on the often hypercapnic and hypoxic habitat of the mudskippers, these fishes may represent an attractive model for pursuing studies on O2 and CO2 sensing due to the air-breathing that increases the importance of acid/base regulation and the O2-related drive including the function of gasotransmitters such as nitric oxide that has an inhibitory (regulatory) function in ionoregulation.

All iguana families with the exception of basilisks share sex chromosomes.

Abstract: Once believed to be restricted only to endotherms (mammals and birds), several poikilothermic amniote lineages have recently been documented to possess long-term evolutionary stability in their sex chromosomes. However, many important lineages were not included in these tests. Previously, based on molecular evidence, we documented the homology of well-differentiated sex chromosomes among seven families of iguanas (Pleurodonta), with basilisks (Corytophanidae) being the only exception, as the tested genes linked to X, but missing on the Y chromosome, in other iguanas were autosomal or pseudoautosomal in basilisks. In this study, we test the homology of sex chromosomes in the remaining, previously unstudied iguana families (Hoplocercidae, Leiosauridae, Liolaemidae, Polychrotidae) and in the basilisk genus Corytophanes. Our results show that 12 currently recognized families of iguanas share X-specific gene content conserved from the common ancestor living in the Cretaceous period. However, the results in the genus Corytophanes indicate the loss of the ancestral differentiated sex chromosomes from the ancestor of basilisks. Our new data further confirm the extensive stability of sex chromosomes in iguanas, thus enabling molecular sexing based on the comparison of the number of X-specific genes by quantitative PCR (qPCR) in all but one family of this widely diversified clade.

Snail mucus - glandular origin and composition in Helix pomatia.

Abstract: Apart from their well-known culinary use, gastropod species such as Helix, which have a hydrogel-like mucus, are increasingly being exploited for cosmetic, bioengineering and medical applications. However, not only are the origin and composition of these "sticky" secretions far from being fully characterized, the number and morphology of the mucus glands involved is also uncertain. This study aims to characterize in detail the cutaneous glands of the Helix pomatia foot on morphological, histochemical and immunohistochemical levels. Hereby the focus is on the gland position and appearance on the foot sole as well as on the chemical nature of the different gland secretions. At least five different gland types can be distinguished by their microanatomy; three are located on the dorsal side and two on the ventral side of the foot sole. Most glands are reactive for acidic proteins and sugars such as mannose and fucose, indicating the presence of acidic glycosaminoglycans. One dorsal gland type shows high reactivity for acidic proteins only. The isolated mucus includes a certain amount of the elements chlorine, potassium and calcium; evidence for lipids was also confirmed in the isolated mucus. The present results for Helix pomatia show a clear difference in the number of glands compared to the related species Helix aspersa (only four mucus glands); histochemically, the glands of both species similarly produce acidic proteins as well as acidic glycosaminoglycans. While calcium ions are known to play a role in mucus formation, the presence and function of other ions such as potassium still need to be clarified.

Patterns of integration in the canine skull: an inside view into the relationship of the skull modules of domestic dogs and wolves.

Abstract: The skull shape variation in domestic dogs exceeds that of grey wolves by far. The artificial selection of dogs has even led to breeds with mismatching upper and lower jaws and maloccluded teeth. For that reason, it has been advocated that their skulls (including the teeth) can be divided into more or less independent modules on the basis of genetics, development or function. In this study, we investigated whether the large diversity of dog skulls and the frequent occurrence of orofacial disproportions can be explained by a lower integration strength between the modules of the skull and by deviations in their covariation pattern when compared to wolves. For that purpose, we employed geometric morphometric methods on the basis of 99 3D-landmarks representing the cranium (subdivided into rostrum and braincase), the mandible (subdivided into ramus and corpus), and the upper and lower tooth rows. These were taken from CT images of 196 dog and wolf skulls. First, we calculated the shape disparity of the mandible and the cranium in dogs and wolves. Then we tested whether the integration strength (measured by RV coefficient) and the covariation pattern (as analysed by partial least squares analysis) of the modules subordinate to the cranium and the mandible can explain differing disparity results. We show, contrary to our expectations, that the higher skull shape diversity in dogs is not explained by less integrated skull modules. Also, the pattern of their covariation in the dog skull can be traced back to similar patterns in the wolf. This shows that existing differences between wolves and dogs are at the utmost a matter of degree and not absolute.

The Ebbinghaus illusion in the gray bamboo shark (Chiloscyllium griseum) in comparison to the teleost damselfish (Chromis chromis).

Abstract: This is the first study to comparatively assess the perception of the Ebbinghaus-Titchener circles and variations of the Delboeuf illusion in four juvenile bamboo sharks (Chiloscyllium griseum) and five damselfish (Chromis chromis) using identical training paradigms We aimed to investigate whether these two species show similarities in the perceptual integration of local elements into the global context The Ebbinghaus-Titchener circles consist of two equally sized central test circles surrounded by smaller or larger circles of different size, number and/or distance During training, sharks and damselfish learned to distinguish a large circle from a small circle, regardless (i) of its gray level and (ii) of the presence of surrounding circles arranged along an outer semi-circle During the subsequent transfer period, individuals were presented with variations of the Ebbinghaus-Titchener circles and the Delboeuf illusion Similar to adult humans, dolphins, or some birds, damselfish tended to judge the test circle surrounded by smaller inducers as larger than the one surrounded by larger inducers (contrast effect) However, sharks significantly preferred the overall larger figure or chose indifferently between both alternatives (assimilation effect) These contrasting responses point towards potential differences in perceptual processing mechanisms, such as 'filling-in' or '(a)modal completion', 'perceptual grouping', and 'local' or 'global' visual perception The present study provides intriguing insights into the perceptual abilities of phylogenetically distant taxa separated in evolutionary time by 200 million years

Shark teeth as edged weapons: serrated teeth of three species of selachians.

Abstract: Prior to European contact, South Pacific islanders used serrated shark teeth as components of tools and weapons. They did this because serrated shark teeth are remarkably effective at slicing through soft tissues. To understand more about the forms and functions of serrated shark teeth, we examined the morphology and histology of tooth serrations in three species: the Tiger Shark (Galeocerdo cuvier), Blue Shark (Prionace glauca), and White Shark (Carcharodon carcharias). We show that there are two basic types of serrations. A primary serration consists of three layers of enameloid with underlying dentine filling the serration's base. All three species studied have primary serrations, although the dentine component differs (orthodentine in Tiger and Blue Sharks; osteodentine in the White Shark). Smaller secondary serrations are found in the Tiger Shark, formed solely by enameloid with no contribution from underlying dentine. Secondary serrations are effectively "serrations within serrations" that allow teeth to cut at different scales. We propose that the cutting edges of Tiger Shark teeth, equipped with serrations at different scales, are linked to a diet that includes large, hard-shelled prey (e.g., sea turtles) as well as smaller, softer prey such as fishes. We discuss other aspects of serration form and function by making analogies to man-made cutting implements, such as knives and saws.

In vivo bone strain and finite element modeling of a rhesus macaque mandible during mastication

Abstract: Finite element analysis (FEA) is a commonly used tool in musculoskeletal biomechanics and vertebrate paleontology. The accuracy and precision of finite element models (FEMs) are reliant on accurate data on bone geometry, muscle forces, boundary conditions and tissue material properties. Simplified modeling assumptions, due to lack of in vivo experimental data on material properties and muscle activation patterns, may introduce analytical errors in analyses where quantitative accuracy is critical for obtaining rigorous results. A subject-specific FEM of a rhesus macaque mandible was constructed, loaded and validated using in vivo data from the same animal. In developing the model, we assessed the impact on model behavior of variation in (i) material properties of the mandibular trabecular bone tissue and teeth; (ii) constraints at the temporomandibular joint and bite point; and (iii) the timing of the muscle activity used to estimate the external forces acting on the model. The best match between the FEA simulation and the in vivo experimental data resulted from modeling the trabecular tissue with an isotropic and homogeneous Young's modulus and Poisson's value of 10GPa and 0.3, respectively; constraining translations along X,Y, Z axes in the chewing (left) side temporomandibular joint, the premolars and the m1; constraining the balancing (right) side temporomandibular joint in the anterior-posterior and superior-inferior axes, and using the muscle force estimated at time of maximum strain magnitude in the lower lateral gauge. The relative strain magnitudes in this model were similar to those recorded in vivo for all strain locations. More detailed analyses of mandibular strain patterns during the power stroke at different times in the chewing cycle are needed.

Functional correlates of the position of the axis of rotation of the mandible during chewing in non-human primates.

Abstract: The location of the axis of rotation (AoR) of the mandible was quantified using the helical axis (HA) in eight individuals from three species of non-human primates: Papio anubis, Cebus apella, and Macaca mulatta. These data were used to test three hypotheses regarding the functional significance of anteroposterior condylar translation - an AoR located inferior to the temporomandibular joint (TMJ) - during chewing: minimizing impingement of the gonial region on cervical soft tissue structures during jaw opening; avoiding stretching of the inferior alveolar neurovascular bundle (IANB); and increasing jaw-elevator muscle torques. The results reveal that the HA is located near the occlusal plane in Papio and Cebus, but closer to the condyle in Macaca; is located anteroinferior to the TMJ during both opening and closing in Papio, as well as during opening in Macaca and Cebus; and varies in its location during closing in Macaca and Cebus. The impingement hypothesis is not supported by interspecific variation in HA location: species with larger gonial angles like Cebus do not have more inferiorly located HAs than species with more obtuse mandibular angles like Papio. However, intraspecific variation provides some support for the impingement hypothesis. The HA seldom passes near or through the lingula, falsifying the hypothesis that its location is determined by the sphenomandibular ligament, and the magnitudes of strain associated with a HA at the TMJ would not be large enough to cause problematic stretching of the IANB. HA location does affect muscle moment arms about the TMJ, with implications for the torque generation capability of the jaw-elevator muscles. In Cebus, a HA farther away from the TMJ is associated with larger jaw-elevator muscle moment arms about the joint than if it were at the TMJ. The effects of HA location on muscle strain and muscle moment arms are largest at large gapes and smallest at low gapes, suggesting that if HA location is of functional significance for primate feeding system performance, it is more likely to be in relation to large gape feeding behaviors than chewing. Its presence in humans is most parsimoniously interpreted as a primitive retention from non-human primate ancestors and explanations for the presence of anteroposterior condylar translation in humans need not invoke either the uniqueness of human speech or upright posture.

Misconceptions about logarithmic transformation and the traditional allometric method.

Abstract: Logarithmic transformation is often assumed to be necessary in allometry to accommodate the kind of variation that accompanies multiplicative growth by plants and animals; and the traditional approach to allometric analysis is commonly believed to have important application even when the bivariate distribution of interest is curvilinear on the logarithmic scale. Here I examine four arguments that have been tendered in support of these perceptions. All the arguments are based on misunderstandings about the traditional method for allometric analysis and/or on a lack of familiarity with newer methods of nonlinear regression. Traditional allometry actually has limited utility because it can be used only to fit a two-parameter power equation that assumes lognormal, heteroscedastic error on the original scale. In contrast, nonlinear regression can fit two- and three-parameter power equations with differing assumptions about structure for error directly to untransformed data. Nonlinear regression should be preferred to the traditional method in future allometric analyses.

Hydric environmental effects on turtle development and sex ratio

Abstract: Experimental and field studies of different turtle species suggest that moisture influences embryonic development and sex ratios, wetter substrates tend to produce more males, and drier substrates produce more females. In this study, we used Trachemys scripta elegans to test the effect of moisture on embryonic development and sex ratios. T. s. elegans eggs were incubated under different temperature and moisture regimes. We monitored embryonic development until stage 22 (after sex determination) and, for the first time, we estimated sex ratios using a male-specific transcriptional molecular marker, Sox9. Among treatments, we found differences in developmental rates, egg mass, and sex ratio. Embryos developed slowly in cooler and wetter sand substrate while water uptake by the eggs was significantly greater on wetter substrates. Developmental differences were due to moisture interacting with temperature where increased water content of the sand resulted in temperatures that were 2-3°C lower than air temperatures. The coolest and the wettest substrates produced 100% males compared to 42% males from the warmest and driest treatment. Further, we found that embryonic growth appears to be more sensitive to temperature at earlier stages of development and to moisture at later stages. This study shows how moisture may change the incubation conditions inside nests by changing the temperature experienced by eggs, which affects development, growth and sex ratios. The results of this study highlight the importance of including moisture conditions when predicting embryo growth and sex ratios and in developing proxies of embryonic development.

Silk gene expression of theridiid spiders: implications for male-specific silk use

Abstract: Spiders (order Araneae) rely on their silks for essential tasks, such as dispersal, prey capture, and reproduction. Spider silks are largely composed of spidroins, members of a protein family that are synthesized in silk glands. As needed, silk stored in silk glands is extruded through spigots on the spinnerets. Nearly all studies of spider silks have been conducted on females; thus, little is known about male silk biology. To shed light on silk use by males, we compared silk gene expression profiles of mature males to those of females from three cob-web weaving species (Theridiidae). We de novo assembled species-specific male transcriptomes from Latrodectus hesperus, Latrodectus geometricus, and Steatoda grossa followed by differential gene expression analyses. Consistent with their complement of silk spigots, male theridiid spiders express appreciable amounts of aciniform, major ampullate, minor ampullate, and pyriform spidroin genes but not tubuliform spidroin genes. The relative expression levels of particular spidroin genes varied between sexes and species. Because mature males desert their prey-capture webs and become cursorial in their search for mates, we anticipated that major ampullate (dragline) spidroin genes would be the silk genes most highly expressed by males. Indeed, major ampullate spidroin genes had the highest expression in S. grossa males. However, minor ampullate spidroin genes were the most highly expressed spidroin genes in L. geometricus and L. hesperus males. Our expression profiling results suggest species-specific adaptive divergence of silk use by male theridiids.

Differences in mating behavior between two allopatric populations of a Neotropical scorpion

Abstract: Courtship and mating behavior generally evolve rapidly in diverging populations and species. The adaptation to different environments may cause behavioral divergence in characteristics involved in mate choice. Our objective in this study was to compare the sexual behavior of two distant populations of the scorpion Bothriurus bonariensis. This species has a broad distribution in South America, inhabiting Central Argentina, Uruguay and south-eastern Brazil. It is known that in this species there is a divergence in morphological patterns (body size, coloration, allometry and fluctuating asymmetry indexes) among distant populations. Considering the differences in environmental conditions between localities, we compare the sexual behavior in intra-population and inter-population matings from Central Argentina and southern Uruguay populations. We found significant differences in mating patterns, including differences in the frequency and duration of important stimulatory courtship behaviors. In addition, most inter-population matings were unsuccessful. In this framework, the differences in reproductive behavior could indicate reproductive isolation between these populations, which coincides with their already known morphological differences. This is the first study comparing the sexual behavior of allopatric populations of scorpions; it provides new data about the degree of intraspecific geographical divergence in the sexual behavior of B. bonariensis.

Functional morphology and evolution of the hyper-elongated intromittent organ in Cassida leaf beetles (Coleoptera: Chrysomelidae: Cassidinae).

Abstract: The peculiar phenomenon of hyper-elongation of intromittent organs is well known in a number of insect groups. However, the unresolved questions of how and why such a phenomenon originated independently many times continue to attract biologists’ attention. To be able to detect the evolutionary driving mechanisms that enabled insects to repeatedly acquire such a peculiarity, first of all the structural key features and the mechanics of these organs have to be determined. In the present study, the morphology of the reproductive organs of two species of the beetle genus Cassida, with a special focus on the male structures, was scrutinised in detail during copulation and at rest using different microscopy techniques. We found that the hyper-elongated structure of the intromittent organ, called flagellum, is part of the male ejaculatory duct. When the flagellum is inserted into the female spermathecal duct, longitudinal muscles of the ejaculatory duct, but not the flagellum, are shortened. These results strongly suggest that the contraction of the longitudinal muscles of the ejaculatory duct causes propulsion of the flagellum into the highly spiralled spermathecal duct of the female. The tip of the cuticular flagellum is curled up, which can suggest that its physical properties differ from those of the rest of the flagellum. Considering the preceding modelling studies, this property aids the flagellum in penetrating within the highly spiralled and very variable female duct. Based on our morphological results and in comparison with the morphology of intromittent organs of other beetles, we discuss the evolutionary origin of the hyper-elongation in the Cassida species and propose a hypothesis that explains the independent origin of the hyper-elongation of intromittent organs.

Physiological differences in preferred temperatures and evaporative water loss rates in two sympatric lacertid species.

Abstract: Sister species living in sympatry offer the opportunity to study the degree of divergence in their ecological, physiological and life-history traits. It has been hypothesized that closely related species with overlapping distribution should differ in their niche to reduce competition for resources. Furthermore, the investigation of sympatric species may shed light on how they may coexist without outcompeting each other. In the present study, we assess the degree of physiological divergence in two sympatric lacertid lizards, Podarcis bocagei and Podarcis guadarramae lusitanicus. These species share a Pliocenic ancestry and overlap at a both geographical and ecological scale. We assessed their thermal preferences and water loss rates, two physiological traits considered stable across congeneric species. We found that the two species differ in both traits, with P. bocagei selecting higher temperature than P. g. lusitanicus and losing more water than the latter at and above its preferred temperature. The results also showed that for both species body size has a relevant impact on thermal and hydric traits, with bigger individuals losing proportionally less water and selecting higher temperatures. These results, combined with previous evidence, suggest that physiological mechanisms, ecological preferences and morphology probably allow these two species to overlap in their distribution while selecting different microhabitats and thus decreasing possible competition between them.

Intracranial and hierarchical perspective on dietary plasticity in mammals

Abstract: The effect of dietary properties on craniofacial form has been the focus of numerous functional studies, with increasingly more work dedicated to the importance of phenotypic plasticity. As bone is a dynamic tissue, morphological variation related to differential loading is well established for many masticatory structures. However, the adaptive osteogenic response of several cranial sites across multiple levels of bony organization remains to be investigated. Here, rabbits were obtained at weaning and raised for 48 weeks until adulthood in order to address the naturalistic influence of altered loading on the long-term development of masticatory and non-masticatory elements. Longitudinal data from micro-computed tomography (μCT) scans were used to test the hypothesis that variation in cortical bone formation and biomineralization in masticatory structures is linked to increased stresses during oral processing of mechanically challenging foods. It was also hypothesized that similar parameters for neurocranial structures would be minimally affected by varying loads as this area is characterized by low strains during mastication and reduced hard-tissue mechanosensitivity. Hypotheses were supported regarding bone formation for maxillomandibular and neurocranial elements, though biomineralization trends of masticatory structures did not mirror macroscale findings. Varying osteogenic responses in masticatory elements suggest that physiological adaptation, and corresponding variation in skeletal performance, may reside differentially at one level of bony architecture, potentially affecting the accuracy of behavioral and in silico reconstructions. Together, these findings underscore the complexity of bone adaptation and highlight functional and developmental variation in determinants of skull form.

Rapid divergence of histones in Hydrozoa (Cnidaria) and evolution of a novel histone involved in DNA damage response in hydra.

Abstract: Histones are fundamental components of chromatin in all eukaryotes. Hydra, an emerging model system belonging to the basal metazoan phylum Cnidaria, provides an ideal platform to understand the evolution of core histone components at the base of eumetazoan phyla. Hydra exhibits peculiar properties such as tremendous regenerative capacity, lack of organismal senescence and rarity of malignancy. In light of the role of histone modifications and histone variants in these processes it is important to understand the nature of histones themselves and their variants in hydra. Here, we report identification of the complete repertoire of histone-coding genes in the Hydra magnipapillata genome. Hydra histones were classified based on their copy numbers, gene structure and other characteristic features. Genomic organization of canonical histone genes revealed the presence of H2A-H2B and H3-H4 paired clusters in high frequency and also a cluster with all core histones along with H1. Phylogenetic analysis of identified members of H2A and H2B histones suggested rapid expansion of these groups in Hydrozoa resulting in the appearance of unique subtypes. Amino acid sequence level comparisons of H2A and H2B forms with bilaterian counterparts suggest the possibility of a highly mobile nature of nucleosomes in hydra. Absolute quantitation of transcripts confirmed the high copy number of histones and supported the canonical nature of H2A. Furthermore, functional characterization of H2A.X.1 and a unique variant H2A.X.2 in the gastric region suggest their role in the maintenance of genome integrity and differentiation processes. These findings provide insights into the evolution of histones and their variants in hydra.

Ontogenetic and functional modularity in the rodent mandible.

Abstract: The material properties of diets consumed by juvenile individuals are known to affect adult morphological outcomes. However, much of the current experimental knowledge regarding dietary effects on masticatory form is derived from studies in which individuals are fed a non-variable diet for the duration of their postweaning growth period. Thus, it remains unclear how intra-individual variation in diet, due to ontogenetic variation in feeding behaviors or seasonal resource fluctuations, affects the resulting adult morphology. Furthermore, the mandible is composed of multiple developmental and functional subunits, and the extent to which growth and plasticity among these modules is correlated may be misestimated by studies that examine non-variable masticatory function in adults only. To address these gaps in our current knowledge, this study raised Sprague Dawley rats (n=42) in four dietary cohorts from weaning to skeletal maturity. Two cohorts were fed a stable ("annual") diet of either solid or powdered pellets. The other two cohorts were fed a variable ("seasonal") diet consisting of solid/powdered pellets for the first half of the study, followed by a shift to the opposite diet. Results of longitudinal morphometric analyses indicate that variation in the mandibular corpus is more prominent at immature ontogenetic stages, likely due to processes of dental eruption and the growth of tooth roots. Furthermore, adult morphology is influenced by both masticatory function and the ontogenetic timing of this function, e.g., the consumption of a mechanically resistant diet. The morphology of the coronoid process was found to separate cohorts on the basis of their early weanling diet, suggesting that the coronoid process/temporalis muscle module may have an early plasticity window related to high growth rates during this life stage. Conversely, the morphology of the angular process was found to be influenced by the consumption of a mechanically resistant diet at any point during the growth period, but with a tendency to reflect the most recent diet. The prolonged plasticity window of the angular process/pterygomasseteric muscle module may be related to delayed ossification and muscular maturation within this module. The research presented here highlights the importance of more naturalistic models of mammalian feeding, and underscores the need for a better understanding of the processes of both morphological and behavioral maturation that follow weaning.

What's in this crab? MRI providing high-resolution three-dimensional insights into recent finds and historical collections of Brachyura.

Abstract: Museum collections may be viewed as a unique window onto the diversity and the functional evolution of species on earth. Detailed information about the inner structure of many precious collectors' items is, however, difficult to gain without destruction of the objects of interest. Here we applied magnetic resonance imaging (MRI) to freshly fixed as well as century-old museum specimens and compared the effects of fixative (formalin, ethanol, mercury chloride) on the image quality. Three-dimensional (3D) reconstruction of MRI was exemplarily used to non-invasively visualize anatomical structures of the brachyuran species Ilia nucleus, Ozius guttatus and Austinograea williamsi. Moreover, the potential of combining MRI and micro-computed tomography (μCT) was exemplarily analyzed for O. guttatus. The best MRI quality was achieved with formalin fixation and this also applied to specimens more than 100 years old. For specimens with a straight carapace width of about 30mm, an isotropic spatial resolution of 100μm allowed for the delineation of all major organ systems such as the nervous system, the gastrointestinal tract, the reproductive system and the heart. Moreover, combining MRI and μCT revealed new insights into the interaction of the heart and surrounding skeletal structures. As examples of its potential, MRI of a specimen of O. guttatus showed a very rare double infection with bopyrid isopods and 3D reconstruction of the reproductive tract of A. williamsi revealed a remarkable size of the ovaries as well as a shape and orientation of the seminal receptacles unusual for brachyurans. Thus, MRI may open up extensive possibilities to study evolutionary and ecological questions by utilizing the immense wealth of natural historical collections without any destruction of the items.

Three-dimensional reconstruction of the embryonic pancreas in the grass snake Natrix natrix L. (Lepidosauria, Serpentes) based on histological studies

Abstract: The aim of this study was to evaluate two research hypotheses: H0-the embryonic pancreas in grass snakes develops in the same manner as in all previously investigated amniotes (from three buds) and its topographical localization within the adult body has no relation to its development; H1-the pancreas develops in a different manner and is related to the different topography of internal organs in snakes. For the evaluation of these hypotheses we used histological methods and three-dimensional (3D) reconstructions of the position of the pancreatic buds and surrounding organs at particular developmental stages and of the final position and shape of the pancreatic gland. Our results indicate that the pancreas primordium in the grass snake is formed by only two buds - a dorsal and a ventral one - that are not connected until the end of stage II. This differs from the majority of vertebrates investigated so far. The gall bladder of the grass snake embryos is connected with the liver only by a thin cystic duct, which also differs from many other vertebrates. Our histological study also indicates a different distribution of the endocrine cells in the embryonic pancreas of the grass snake because the first endocrine cells appeared in the dorsal part of the pancreas in a region located close to the spleen. During the entire developmental period no evidence of these cells was found in the ventral part of the pancreas. The endocrine cells form elongated, large and irregular-shaped islets. They can also form structures resembling "inverted acini". Such an arrangement is characteristic of snakes only. The differentiating pancreas penetrates the ventral part of the developing spleen and divides it into three separate parts at developmental stage IX. This is unique among vertebrates. At the end of the embryonic development (stage XI), the pancreas, the spleen and the gall bladder are located in close proximity and form the so-called triad. Our results suggest that the untypical topography of the organ systems in snakes may determine the unique development of the pancreas in these animals.

Functional trade-offs in the aquatic feeding performance of salamanders.

Abstract: During aquatic feeding salamanders use the hyobranchial apparatus to capture prey. The hyobranchial apparatus depresses the floor of the mouth, effectively expanding the oropharyngeal cavity and generating suction. Within the family Salamandridae, there is a wide range of ecological diversity, with salamanders being terrestrial, semi-aquatic, or aquatic as adults. The purpose of this research was to quantify the diverse morphology and suction feeding performance of aquatically feeding salamandrids. We hypothesized that a more robust hyobranchial apparatus morphology would yield increased aquatic feeding performance. When compared to semi-aquatic newts, the fully aquatic species Paramesotriton labiatus had greater mineralization of the hyobranchial apparatus, as well as relatively more narrow basibranchial and wider ceratobranchial I+II complexes. These morphological differences coincide with greater aquatic feeding performance. Kinematics from high-speed videography revealed that maximum mouth opening velocity and acceleration were approximately two and five times greater, respectively, in Paramesotriton, and hyobranchial depression acceleration was found to be approximately three times greater than in the semi-aquatic species Pleurodeles waltl, Notophthalmus viridescens, Triturus dobrogicus, and Cynops cyanurus. Using digital particle image velocimetry, peak and average fluid velocity generated in Paramesotriton during suction feeding events were found to be 0.5ms-1 and 0.2ms-1, respectively, doubling that of all semi-aquatic species. These findings reveal that specialized morphology increases aquatic feeding performance in a fully aquatic newt.

Gross anatomy and histology of the olfactory rosette of the shark Heptranchias perlo

Abstract: Sharks belonging to the family Hexanchidae have six or seven gill slits, unlike all other elasmobranchs, which have five gill slits. Their olfactory organs have a round shape, which is common for holocephalans, but not for elasmobranchs. Thus, the shape of the olfactory organ represents a further, less striking, peculiarity of this family among elasmobranchs. Despite that, the microscopic anatomy and histology of the olfactory organ have not yet been studied in any species of this family. Here, an anatomical and histological description of the olfactory organ of the sharpnose sevengill shark Heptranchias perlo is given. The organ is a rosette, with a central raphe and 31-34 primary lamellae, which bear secondary lamellae with a more or less branched shape. The elastic connective capsule which envelops the olfactory rosette possibly changes its shape along with water influx. In the olfactory epithelium, the supporting cells also have a secretory function, while no specialized mucous cells are visible; regarding this feature the olfactory epithelium of H. perlo differs from that of other chondrichthyan species. The immunohistochemical investigation of the sensory epithelium shows the absence of immunoreactivity for Gαolf in receptor neurons, which confirms previous observations in Chondrichthyes.

Diet of Amazon river turtles (Podocnemididae): a review of the effects of body size, phylogeny, season and habitat.

Abstract: Amazon rivers can be divided into three groups (black, white and clear waters) according to the origin of their sediment, dissolved nutrient content, and vegetation. White water rivers have high sediment loads and primary productivity, with abundant aquatic and terrestrial plant life. In contrast, black water rivers are acid and nutrient-poor, with infertile floodplains that support plant species exceptionally rich in secondary chemical defences against herbivory. In this study, we reviewed available information on the diet of Amazon sideneck river turtles (Family Podocnemididae). Our aim was to test the relationship between water type and diet of podocnemidids. We also took into account the effects of season, size, age, sex and phylogeny. Based on our review, turtles of this family are primarily herbivorous but opportunistic, consuming from 46 to 99% (percent volume) of vegetable matter depending on species, sex, season and location. There was no significant correlation between the maximum carapace size of a species and vegetable matter consumed. When the available information on diet, size and habitat was arranged on the podocnemidid phylogeny, no obvious evolutionary trend was evident. The physicochemical properties of the inhabited water type indirectly influence the average volume of total vegetable matter consumed. Species with no specialised stomach adaptations for herbivory consumed smaller amounts of hard to digest vegetable matter (i.e. leaves, shoots and stems). We propose that turtles with specialized digestive tracts may have an advantage in black water rivers where plant chemical defences are more common. Despite limitations of the published data our review highlights the overall pattern of diet in the Podocnemididae and flags areas where more studies are needed.

Inter-sexual habitat and isotopic niche segregation of the endangered Monteiro's storm-petrel during breeding.

Abstract: At-sea distribution and trophic ecology of small seabird species (i.e.<100 g) is far less known when compared to their larger relatives. We studied the habitat use (spatial ecology) and isotopic niches (trophic ecology) of the endangered Monteiro's storm-petrel Hydrobates monteiroi during the incubation and chick-rearing periods of 2013. There was a sexual foraging segregation of Monteiro's storm-petrels during the breeding period (tracking data) but also during the non-breeding stage (stable isotope analysis). Females took advantage of their longer wings to forage over the shallower Mid-Atlantic ridge (MAR) north of Azores, under colder and windier regimes when compared to males, who mostly exploited northern deep waters comparatively closer to the breeding colony. Between-sex differences in the spatial distribution were more obvious during the incubation period, with the overlap in their distribution increasing during the chick-rearing phase. There was also an isotopic segregation between sexes both during the previous breeding and the non-breeding stages, with females exhibiting a narrower, lower level isotopic niche when compared to males. Though the distribution patterns reported here should be useful for the at-sea conservation of this endangered species, future research should focus on (1) performing year-round tracking to map the species' distribution during the non-breeding period and (2) gathering multi-year tracking information to understand the effect of inter-annual environmental stochasticity on the foraging choices and trophic habits of the species.

Interaction between visual and chemical cues in a Liolaemus lizard: a multimodal approach.

Abstract: Multimodal communication involves the use of signals and cues across two or more sensory modalities. The genus Liolaemus (Iguania: Liolaemidae) offers a great potential for studies on the ecology and evolution of multimodal communication, including visual and chemical signals. In this study, we analyzed the response of male and female Liolaemus pacha to chemical, visual and combined (multimodal) stimuli. Using cue-isolation tests, we registered the number of tongue flicks and headbob displays from exposure to signals in each modality. Number of tongue flicks was greater when a chemical stimulus was presented alone than in the presence of visual or multimodal stimuli. In contrast, headbob displays were fewer in number with visual and chemical stimuli alone, but significantly higher in number when combined. Female signallers triggered significantly more tongue flicks than male signallers, suggesting that chemical cues are involved in sexual recognition. We did not find an inhibition between chemical and visual cues. On the contrary, we observed a dominance of the chemical modality, because when presented with visual stimuli, lizards also responded with more tongue flicks than headbob displays. The total response produced by multimodal stimuli was similar to that of the chemical stimuli alone, possibly suggesting non-redundancy. We discuss whether the visual component of a multimodal signal could attract attention at a distance, increasing the effectiveness of transmission and reception of the information in chemical cues.

Vascular distensibilities have minor effects on intracardiac shunt patterns in reptiles.

Abstract: The different vascular distensibilities of systemic and pulmonary circuits were recently proposed as an important mechanism defining the direction of cardiac shunt and the distribution of cardiac output in vertebrates with undivided cardiac ventricles. In short, the more distensible pulmonary vascular bed was proposed to accommodate a larger portion of the blood ejected from the heart when cardiac output increases. To evaluate this hypothesis, we performed a meta-analysis based on fourteen previously published studies in two species of reptiles (the freshwater turtle, Trachemys scripta, and the South American rattlesnake, Crotalus durissus) to describe how cardiac shunt patterns change when cardiac output increases. We found no general linear relation between total blood flow and systemic or pulmonary blood flows. Thus, cardiac output alterations cannot be used as a reliable reference for prediction of shunt patterns in both T. scripta and C. durissus. Hence, differential distensibilities appear to be of minor importance for the determination of cardiac shunt patterns in the species analyzed. We speculate that the lack of correlation in these phylogenetically distant species may indicate that this is a common trend within non-crocodilian reptiles.

Interplay between behavior, morphology and physiology supports lepidophagy in the catfish Pachypterus khavalchor (Siluriformes: Horabagridae).

Abstract: The present study demonstrates the scale-eating behavior of a siluroid catfish, Pachypterus khavalchor, and the role of its oral structure and bacterial endosymbionts in shaping this lepidophagous habit. Scale-eating behavior in P. khavalchor was studied using a series of behavioral experiments. P. khavalchor was found to feed only on the scales of live fish and never of dead fish, even after 72h of starvation. It was nocturnal in habit and attacked all species that were used as prey. Attacking behavior showed extensive chasing of prey species followed by a powerful random strike at the flank or close to the caudal region in posterior oblique position. After a strike, P. khavalchor was found to immediately turn back and pick up the falling dislodged scales. SEM analysis of oral structures of P. khavalchor revealed three different types of teeth arranged in specific order on the upper jaw, lower jaw and pharyngeal region that could facilitate their lepidophagous habit. Teeth of upper and lower jaws are likely to help in dislodging the scales and pharyngeal teeth may help to engulf the scales. Gut microflora analysis and enzyme assay revealed two isolates, namely Bacillus pumilus and Bacillus licheniformis, which were positive for chitinase production. These two isolates were found to be capable of producing chitinase indicating that they are likely to be involved in the digestion of chitin-rich scales in the host fish gut. An in vitro scale degradation assay further strengthens the results since both isolates were found to be efficient in chitinase production and degradation of scales.

Resource allocation and post-reproductive degeneration in the freshwater cnidarian Hydra oligactis (Pallas, 1766).

Abstract: Freshwater hydra are among the few animal groups that show negligible senescence and can maintain high survival and reproduction rates when kept under stable conditions in the laboratory. Yet, one species of Hydra (H. oligactis) undergoes a senescence-like process in which polyps degenerate and die after sexual reproduction. The ultimate factors responsible for this phenomenon are unclear. High mortality in reproducing animals could be the consequence of increased allocation of resources to reproduction at the expense of somatic maintenance. This hypothesis predicts that patterns of reproduction and survival are influenced by resource availability. To test this prediction we investigated survival and reproduction at different levels of food availability in 10 lineages of H. oligactis derived from a single Hungarian population. Sexual reproduction was accompanied by reduced survival, but a substantial proportion of animals regenerated after sexual reproduction and continued reproducing asexually. Polyps belonging to different lineages showed differences in their propensity to initiate sexual reproduction, gonad number and survival rate. Food availability significantly affected fecundity (number of eggs or testes produced), with the largest number of gonads being produced by animals kept on a high food regime. On the other hand, survival rate was not affected by the amount of food. These results show that survival is conserved at the expense of reproduction in this population when food is low. It remains a question still to be answered why survival is prioritized over reproduction in this population.