Power matters in closing the phenotyping gap

Much of our understanding of physiology and metabolism is derived from investigating mouse mutants and transgenic mice, and open-access platforms for standardized mouse phenotyping such as the German Mouse Clinic (GMC) are currently viewed as one powerful tool for identifying novel gene-function relationships. Phenotyping or phenotypic screening involves the comparison of wild-type control mice with their mutant or transgenic littermates. In our study, we explored the extent to which standardized phenotyping will succeed in detecting biologically relevant phenotypic differences in mice generated and provided by different collaborators. We analyzed quantitative metabolic data (body mass, energy intake, and energy metabolized) collected at the GMC under the current workflow, and used them for statistical power considerations. Our results demonstrate that there is substantial variability in these parameters among lines of wild-type C57BL/6 (B6) mice from different sources. Given this variable background noise in mice that serve as controls, subtle phenotypes in mutant or transgenic littermates may be overlooked. Furthermore, a phenotype observed in one cohort of a mutant line may not be reproducible (to the same extent) in mice coming from a different environment or supplier. In the light of these constraints, we encourage researchers to incorporate information on intrastrain variability into future study planning, or to perform advanced hierarchical analyses. Both will ultimately improve the detectability of novel phenotypes by phenotypic screening. Carola W. Meyer and Ralf Elvert contributed equally to this work.     

Sperm bundle and reproductive organs of carabid beetles tribe Pterostichini (Coleoptera: Carabidae)

The morphological characteristics of sperm and reproductive organs may offer clues as to how reproductive systems have evolved. In this paper, the morphologies of the sperm and male reproductive organs of carabid beetles in the tribe Pterostichini (Coleoptera: Carabidae) are described, and the morphological associations among characters are examined. All species form sperm bundles in which the head of the sperm was embedded in a rod-shaped structure, i.e., spermatodesm. The spermatodesm shape (left-handed spiral, right-handed spiral, or without conspicuous spiral structure) and the condition of the sperm on the spermatodesm surface (with the tail free-moving or forming a thin, sheetlike structure) vary among species. In all species, the spiral directions of the convoluted seminal vesicles and vasa deferentia are the same on both sides of the body; that is, they show an asymmetric structure. The species in which the sperm bundle and the seminal vesicles both have a spiral structure could be classified into two types, with significant differences in sperm-bundle length between the two types. The species with a sperm-bundle spiral and seminal-vesicle spiral of almost the same diameter have longer sperm bundles than the species with a sperm-bundle spiral and seminal-vesicle tube of almost the same diameter. In the former type, the spiral directions of the sperm bundles and seminal vesicles are inevitably the same, whereas they differ in some species with the later type. Therefore, increased sperm bundle length appears to have been facilitated by the concordance of the sperm bundle’s coiling direction with the coiling direction of the seminal vesicle.     

Echolocation calls of Poey’s flower bat (<Emphasis Type="Italic">Phyllonycteris poeyi</Emphasis>) unlike those of other phyllostomids

Unlike any other foraging phyllostomid bat studied to date, Poey’s flower bats (Phyllonycteris poeyi-Phyllostomidae) emit relatively long (up to 7.2 ms), intense, single-harmonic echolocation calls. These calls are readily detectable at distances of at least 15 m. Furthermore, the echolocation calls contain only the first harmonic, which is usually filtered out in the vocal tract of phyllostomids. The foraging echolocation calls of P. poeyi are more like search-phase echolocation calls of sympatric aerial-feeding bats (Molossidae, Vespertilionidae, Mormoopidae). Intense, long, narrowband, single-harmonic echolocation calls focus acoustic energy maximizing range and favoring detection, which may be particularly important for cruising bats, like P. poeyi, when flying in the open. Flying in enclosed spaces, P. poeyi emit short, low-intensity, frequency-modulated, multiharmonic echolocation calls typical of other phyllostomids. This is the first report of a phyllostomid species emitting long, intense, single-harmonic echolocation calls with most energy in the first harmonic.     

Fossil insect evidence for the end of the Western Settlement in Norse Greenland

The fate of Norse farming settlements in southwest Greenland has often been seen as one of the great mysteries of North Atlantic colonization and expansion. Preservation of organic remains in the permafrost of the area of the Western Settlement, inland from the modern capital Nuuk, allowed very detailed study of the phases of occupation. Samples were taken from house floors and middens during the process of archaeological excavations and from insect remains were abstracted and identified in the laboratory. In this study, we present a new paleoecological approach principally examining the fossil fly faunas from house floors. The results of our study provide contrasting detailed pictures of the demise of two neighboring farms, Gården under Sandet and Nipaatsoq, one where abandonment appears as part of a normal process of site selection and desertion, and the other where the end was more traumatic. The level of detail, which was obtained by analysis of the dipterous (true fly) remains, exceeds all previous work and provides insights otherwise unobtainable.     

Last Glacial mammals in South America: a new scenario from the Tarija Basin (Bolivia)

The chronology, sedimentary history, and paleoecology of the Tarija Basin (Bolivia), one of the richest Pleistocene mammalian sites in South America, are revised here based on a multidisciplinary study, including stratigraphy, sedimentology, geomorphology, paleontology, isotope geochemistry, and ¹⁴C geochronology. Previous studies have indicated a Middle Pleistocene age for this classic locality. We have been able to obtain a series of ¹⁴C dates encompassing all the fossil-bearing sequences previously studied in the Tarija Basin. The dated layers range in age from about 44,000 to 21,000 radiocarbon years before present (BP), indicating that the Tarija fauna is much younger than previously thought. Glacial advances correlated to marine isotopic stages (MIS) 4 and 2 (ca. 62 and 20 ka BP, respectively) are also documented at the base and at the very top of the Tarija–Padcaya succession, respectively, indicating that the Bolivian Altiplano was not dry but sustained an ice cap during the Last Glacial Maximum. The results of this multidisciplinary study enable us to redefine the chronological limits of the Tarija sequence and of its faunal assemblage and to shift this paleontological, paleoclimatological, and paleoecological framework to the time interval from MIS 4 to MIS 2.     

Spiroplasma infection causes either early or late male killing in <Emphasis Type="Italic">Drosophila</Emphasis>, depending on maternal host age

Symbiont-induced male-killing phenotypes have been found in a variety of insects. Conventionally, these phenotypes have been divided into two categories according to the timing of action: early male killing at embryonic stages and late male killing at late larval stages. In Drosophila species, endosymbiotic bacteria of the genus Spiroplasma have been known to cause early male killing. Here, we report that a spiroplasma strain normally causing early male killing also induces late male killing depending on the maternal host age: male-specific mortality of larvae and pupae was more frequently observed in the offspring of young females. As the lowest spiroplasma density and occasional male production were also associated with newly emerged females, we proposed the density-dependent hypothesis for the expression of early and late male-killing phenotypes. Our finding suggested that (1) early and late male-killing phenotypes can be caused by the same symbiont and probably by the same mechanism; (2) late male killing may occur as an attenuated expression of early male killing; (3) expression of early and late male-killing phenotypes may be dependent on the symbiont density, and thus, could potentially be affected by the host immunity and regulation; and (4) early male killing and late male killing could be alternative strategies adopted by microbial reproductive manipulators.     

A male gift to its partner? Cyanogenic glycosides in the spermatophore of longwing butterflies (<Emphasis Type="Italic">Heliconius</Emphasis>)

Males of several insect species transfer nuptial gifts to females during mating, typically in the form of a protein-rich spermatophore. In chemically defended species, males could potentially enhance such a gift with chemicals that help protect the female, her eggs, or both. This was shown for lepidopteran species that accumulate pyrrolizidine alkaloids. Most Heliconius butterflies are presumably protected from predators by virtue of de novo synthesized and/or sequestered cyanogenic glycosides. Males of Heliconius species are known to transfer nutritional gifts to the females but whether defensive chemicals could also be transferred is not known. To ascertain whether transfer of cyanogens occurs, we dissected freshly mated females from nine different Heliconius species and analyzed spermatophores for cyanogenic glycosides. We found cyanogens in the spermatophores of all nine species. This is the first time cyanogenic glycosides are reported in the spermatophores of arthropods. We discuss the implications of these findings for Heliconius biology and for other cyanogenic insects as well. We suggest that chemically defended species commonly lace their nuptial gifts with defensive chemicals to improve gift quality.     

Soil and preen waxes influence the expression of carotenoid-based plumage coloration

The signaling function of carotenoid-based plumage is mainly determined by the concentration of pigments in feathers. For this reason, most studies of the proximate control of coloration focus on processes during and preceding moult. In great tits Parus major, past research demonstrates that carotenoid-based plumage coloration honestly indicates male quality and, thus, may be a sexually selected signal. In this study, we investigate how dirt and preen oil influence the coloration of carotenoid-based feathers in the great tit. We collected six feathers from each individual bird; three feathers served as controls while the remaining three feathers were washed with a chloroform/methanol mixture to remove soil and preen waxes. We assessed plumage coloration using digital photography. This cleaning procedure slightly enhanced ornamentation; the experimentally cleaned feathers expressed hues shifted towards shorter wavelengths and expressed brighter overall coloration than control feathers. This is the first experimental study conducted on wild birds demonstrating that, in addition to pigment concentration, the presence of preen waxes and soils on feathers may contribute to variation in coloration.     

Body mass estimations for <Emphasis Type="Italic">Plateosaurus engelhardti</Emphasis> using laser scanning and 3D reconstruction methods

Both body mass and surface area are factors determining the essence of any living organism. This should also hold true for an extinct organism such as a dinosaur. The present report discusses the use of a new 3D laser scanner method to establish body masses and surface areas of an Asian elephant (Zoological Museum of Copenhagen, Denmark) and of Plateosaurus engelhardti, a prosauropod from the Upper Triassic, exhibited at the Paleontological Museum in Tübingen (Germany). This method was used to study the effect that slight changes in body shape had on body mass for P. engelhardti. It was established that body volumes varied between 0.79 m³ (slim version) and 1.14 m³ (robust version), resulting in a presumable body mass of 630 and 912 kg, respectively. The total body surface areas ranged between 8.8 and 10.2 m², of which, in both reconstructions of P. engelhardti, ∼33% account for the thorax area alone. The main difference between the two models is in the tail and hind limb reconstruction. The tail of the slim version has a surface area of 1.98 m², whereas that of the robust version has a surface area of 2.73 m². The body volumes calculated for the slim version were as follows: head 0.006 m³, neck 0.016 m³, fore limbs 0.020 m³, hind limbs 0.08 m³, thoracic cavity 0.533 m³, and tail 0.136 m³. For the robust model, the following volumes were established: 0.01 m³ head, neck 0.026 m³, fore limbs 0.025 m³, hind limbs 0.18 m³, thoracic cavity 0.616 m³, and finally, tail 0.28 m³. Based on these body volumes, scaling equations were used to assess the size that the organs of this extinct dinosaur have.