Multifluorescence High‐Resolution Episcopic Microscopy for 3D Imaging of Adult Murine Organs

3D microscopy of large biological samples (>0.5 cm^{3})is transforming biological research. Many existing techniques require trade-offs between image resolution, sample size, and method complexity. A simple robust instrument with the potential to conduct large-volume 3D imaging currently exists in the form of the optical high-resolution episcopic microscopy (HREM). However, the development of the instrument to date is limited to single-fluorescent wavelength imaging with nonspecific eosin staining. Herein, developments to realize the potential of the HREM to become multifluorescent high-resolution episcopic microscopy (MF-HREM) are presented. MF-HREM is a serial-sectioning and block-facing wide-field fluorescence imaging technique, which does not require tissue clearing or optical sectioning. Multiple developments are detailed in sample preparation and image postprocessing to enable multiple specific stains in large samples and show how these enable segmentation and quantification of the data. The application of MF-HREM is demonstrated in a variety of biological contexts: 3D imaging of whole tumor vascular networks and tumor cell invasion in xenograft tumors up to 7.5 mm^{3} at resolutions of 2.75 μm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white-matter track orientation in adult mouse brain

Examination of Clock and Adcyap1 gene variation in a neotropical migratory passerine

Complex behavioral traits, such as those making up a migratory phenotype, are regulated by multiple environmental factors and multiple genes. We investigated possible relationships between microsatellite variation at two candidate genes implicated in the control of migratory behavior, Clock and Adcyap1, and several aspects of migratory life-history and evolutionary divergence in the Painted Bunting (Passerina ciris), a species that shows wide variation in migratory and molting strategies across a disjunct distribution. We focused on Clock and Adcyap1 microsatellite variation across three Painted Bunting populations in Oklahoma, Louisiana, and North Carolina, and for the Oklahoma breeding population we used published migration tracking data on adult males to explore phenotypic variation in individual migratory behavior. We found no correlation between microsatellite allele size within either Clock and Adcyap1 relative to the initiation or duration of fall migration in adult males breeding in Oklahoma. We also show the lack of significant correlations with aspects of the migratory phenotype for the Louisiana population. Our research highlights the limitations of studying microsatellite allelic mutations that are of undetermined functional influence relative to complex behavioral phenotypes.This research was funded by the National Science Foundation (grants nos. IDBR 1014891 and DEB 0946685) and by the United States Department of Agriculture (grant no. NIFA-AFRI-003536). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.YesEach submission to PLOS ONE passes through a rigorous quality control and peer-review evaluation process before receiving a decision. The initial in-house quality control check deals with issues such as competing interests; ethical requirements for studies involving human participants or animals; financial disclosures; full compliance with PLOS’ data availability policy, etc. Submissions may be returned to authors for queries, and will not be seen by our Editorial Board or peer reviewers until they pass this quality control check. Once each manuscript has passed quality control, it is assigned to a member of the Editorial Board, who takes responsibility as the Academic Editor for the submission. The Academic Editor is responsible for conducting the peer-review process and for making a decision to accept, invite revision of, or reject the article

Perceptions of animal physiotherapy amongst Irish veterinary surgeons

The aim of this study was to investigate veterinary surgeons' perceptions, knowledge and use of animal physiotherapy in the Republic of Ireland. A questionnaire was developed and sent to 200 veterinary surgeons, of which 97 were returned. Results indicated that 77 (79%) of respondents were aware of animal physiotherapists. Common sources of information included veterinary colleagues, owners and professional journals, with physiotherapists themselves and undergraduate training being less commonly cited. Awareness of animal physiotherapy was greatest amongst those working in equine practice (χ2 = 5.7, df 1, p = 0.017); they were more knowledgeable about its techniques (t = 2.806, df 75, p = 0.006) and more likely to refer (χ2 = 48.36, df 1, p = 0.0001). Seventy-four respondents (96%) thought that more research was necessary to increase the evidence base for animal physiotherapy. If this branch of physiotherapy is to develop, there needs to be increased interaction and co-operation between veterinary surgeons and chartered animal physiotherapists

Multifluorescence High-Resolution Episcopic Microscopy for 3D Imaging of Adult Murine Organs

3D microscopy of large biological samples (>0.5 cm3) is transforming biological research. Many existing techniques require trade‐offs between image resolution, sample size, and method complexity. A simple robust instrument with the potential to conduct large‐volume 3D imaging currently exists in the form of the optical high‐resolution episcopic microscopy (HREM). However, the development of the instrument to date is limited to single‐fluorescent wavelength imaging with nonspecific eosin staining. Herein, developments to realize the potential of the HREM to become multifluorescent high‐resolution episcopic microscopy (MF‐HREM) are presented. MF‐HREM is a serial‐sectioning and block‐facing wide‐field fluorescence imaging technique, which does not require tissue clearing or optical sectioning. Multiple developments are detailed in sample preparation and image postprocessing to enable multiple specific stains in large samples and show how these enable segmentation and quantification of the data. The application of MF‐HREM is demonstrated in a variety of biological contexts: 3D imaging of whole tumor vascular networks and tumor cell invasion in xenograft tumors up to 7.5 mm3 at resolutions of 2.75 μm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white‐matter track orientation in adult mouse brain

Full lifetime perspectives on the costs and benefits of lay date variation in tree swallows

Animals must balance various costs and benefits when deciding when to breed. The costs and benefits of breeding at different times have received much attention, but most studies have been limited to investigating short-term season-to-season fitness effects. However, breeding early, versus late, in a season may influence lifetime fitness over many years, trading off in complex ways across the breeder?s lifepan. In this study, we examined the complete life histories of 867 female tree swallows (Tachycineta bicolor) breeding in Ithaca, New York, between 2002 and 2016. Earlier breeders outperformed later breeders in short-term measures of reproductive output and offspring quality. Though there were weak indications that females paid long-term future survival costs for breeding early, lifetime fledgling output was markedly higher overall in early-breeding birds. Importantly, older females breeding later in the season did not experience compensating life-history advantages that suggested an alternative equal-fitness breeding strategy. Rather, most or all of the swallows appear to be breeding as early as they can, and differences in lay dates appear to be determined primarily by differences in individual quality or condition. Lay date had a significant repeatability across breeding attempts by the same female, and the first lay date of females fledged in our population was strongly influenced by the first lay date of their mothers, indicating the potential for ongoing selection on lay date. By examining performance over the entire lifespan of a large number of individuals, we were able to clarify the relationship between timing of breeding and fitness and gain new insight into the sources of variability in this important life history trait.Fil: Winkler, David Ward. Cornell University; Estados UnidosFil: Hallinger, Kelly K.. Cornell University; Estados UnidosFil: Pegan, Teresa M.. University of Michigan; Estados UnidosFil: Taff, Conor C.. Cornell University; Estados UnidosFil: Verhoeven, Mo A.. University of Groningen; Países BajosFil: Van Oordt, David Chang. Cornell University; Estados UnidosFil: Stager, Maria. University of Montana; Estados UnidosFil: Uehling, Jennifer J.. Cornell University; Estados UnidosFil: Vitousek, Maren N.. Cornell University; Estados UnidosFil: Andersen, Michael J.. University of New Mexico; Estados UnidosFil: Ardia, Daniel R.. Franklin & Marshall College; Estados UnidosFil: Belmaker, Amos. Tel Aviv University; IsraelFil: Ferretti, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Forsman, Anna M.. University Of Central Florida; Estados UnidosFil: Gaul, Jennifer R.. International High School at La Guardia Community College; Estados UnidosFil: Llambias, Paulo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Orzechowski, Sophia C.. Harvard University; Estados UnidosFil: Shipley, Ryan. Max Planck Institute For Animal Behavior; AlemaniaFil: Wilson, Maya. Virginia Polytechnic Institute. Department Of Geological Sciences; Estados UnidosFil: Yoon, Hyun Seok. University of Tennessee; Estados Unido

Pulsed subduction accretion and tectonic erosion reconstructed since 2.5 Ma from the tephra record offshore Costa Rica

Author Posting. © American Geophysical Union, 2005. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geochemistry Geophysics Geosystems 6 (2005): Q09016, doi:10.1029/2005GC000963.Tephra layers recovered by Ocean Drilling Program from the forearc and trench regions offshore the Nicoya Peninsula of Costa Rica allow the temporal evolution of the volcanic arc to be reconstructed since 2.5 Ma. Major and trace element analyses by microprobe methods reveal a dominant tholeiitic character and a provenance in the Costa Rican area. The tephra show long-term coherent variability in geochemistry. One tephra dated at 1.45 Ma shows minimum values in ɛ Nd and maximum Li/Y consistent with very high degrees of sediment recycling at this time. However, overall Li/Y and δ7Li increase with SiO2 content, suggesting addition of heavy Li through forearc tectonic erosion and crustal assimilation. Peak values in δ7Li starting at 1.45 Ma and lasting ∼0.5 m.y. indicate enhanced tectonic erosion of the forearc possibly caused by subduction of a seamount at 1.45 Ma. The tephra record indicates significant temporal variability in terms of sediment subduction, reconciling the geologic evidence for long-term tectonic erosion and geochemical evidence for recent sediment accretion in the modern Central American arc.Financial support for the analytical work was gratefully received from JOI-USSAC. The lithium isotope work was supported in part by National Science Foundation grant OCE-990554 to L.H.C

Linking human impacts to community processes in terrestrial and freshwater ecosystems

Human impacts such as habitat loss, climate change and biological invasions are radically altering biodiversity, with greater effects projected into the future. Evidence suggests human impacts may differ substantially between terrestrial and freshwater ecosystems, but the reasons for these differences are poorly understood. We propose an integrative approach to explain these differences by linking impacts to four fundamental processes that structure communities: dispersal, speciation, species-level selection and ecological drift. Our goal is to provide process-based insights into why human impacts, and responses to impacts, may differ across ecosystem types using a mechanistic, eco-evolutionary comparative framework. To enable these insights, we review and synthesise (i) how the four processes influence diversity and dynamics in terrestrial versus freshwater communities, specifically whether the relative importance of each process differs among ecosystems, and (ii) the pathways by which human impacts can produce divergent responses across ecosystems, due to differences in the strength of processes among ecosystems we identify. Finally, we highlight research gaps and next steps, and discuss how this approach can provide new insights for conservation. By focusing on the processes that shape diversity in communities, we aim to mechanistically link human impacts to ongoing and future changes in ecosystems

Linking human impacts to community processes in terrestrial and freshwater ecosystems.

Human impacts such as habitat loss, climate change and biological invasions are radically altering biodiversity, with greater effects projected into the future. Evidence suggests human impacts may differ substantially between terrestrial and freshwater ecosystems, but the reasons for these differences are poorly understood. We propose an integrative approach to explain these differences by linking impacts to four fundamental processes that structure communities: dispersal, speciation, species-level selection and ecological drift. Our goal is to provide process-based insights into why human impacts, and responses to impacts, may differ across ecosystem types using a mechanistic, eco-evolutionary comparative framework. To enable these insights, we review and synthesise (i) how the four processes influence diversity and dynamics in terrestrial versus freshwater communities, specifically whether the relative importance of each process differs among ecosystems, and (ii) the pathways by which human impacts can produce divergent responses across ecosystems, due to differences in the strength of processes among ecosystems we identify. Finally, we highlight research gaps and next steps, and discuss how this approach can provide new insights for conservation. By focusing on the processes that shape diversity in communities, we aim to mechanistically link human impacts to ongoing and future changes in ecosystems