Geolocators lead to better measures of timing and renesting in black-tailed godwits and reveal the bias of traditional observational methods

Long-term population studies can identify changes in population dynamics over time. However, to realize meaningful conclusions, these studies rely on accurate measurements of individual traits and population characteristics. Here, we evaluate the accuracy of the observational methods used to measure reproductive traits in individually marked black-tailed godwits (Limosa limosa limosa). By comparing estimates from traditional methods with data obtained from light-level geolocators, we provide an accurate estimate of the likelihood of renesting in godwits and the repeatability of the lay dates of first clutches. From 2012 to 2018, we used periods of shading recorded on the light-level geolocators carried by 68 individual godwits to document their nesting behaviour. We then compared these estimates to those simultaneously obtained by our long-term observational study. We found that among recaptured geolocator-carrying godwits, all birds renested after a failed first clutch, regardless of the date of nest loss or the number of days already spent incubating. We also found that 43% of these godwits laid a second replacement clutch after a failed first replacement, and that 21% of these godwits renested after a hatched first clutch. However, the observational study correctly identified only 3% of the replacement clutches produced by geolocator-carrying individuals and designated as first clutches a number of nests that were actually replacement clutches. Additionally, on the basis of the observational study, the repeatability of lay date was 0.24 (95% CI 0.17–0.31), whereas it was 0.54 (95% CI 0.28–0.75) using geolocator-carrying individuals. We use examples from our own and other godwit studies to illustrate how the biases in our observational study discovered here may have affected the outcome of demographic estimates, individual-level comparisons, and the design, implementation and evaluation of conservation practices. These examples emphasize the importance of improving and validating field methodologies and show how the addition of new tools can be transformational.     

Microarchitecture Is Severely Compromised but Motor Protein Function Is Preserved in Dystrophic mdx Skeletal Muscle

Progressive force loss in Duchenne muscular dystrophy is characterized by degeneration/regeneration cycles and fibrosis. Disease progression may involve structural remodeling of muscle tissue. An effect on molecular motorprotein function may also be possible. We used second harmonic generation imaging to reveal vastly altered subcellular sarcomere microarchitecture in intact single dystrophic mdx muscle cells (∼1 year old). Myofibril tilting, twisting, and local axis deviations explain at least up to 20% of force drop during unsynchronized contractile activation as judged from cosine angle sums of myofibril orientations within mdx fibers. In contrast, in vitro motility assays showed unaltered sliding velocities of single mdx fiber myosin extracts. Closer quantification of the microarchitecture revealed that dystrophic fibers had significantly more Y-shaped sarcomere irregularities (“verniers”) than wild-type fibers (∼130/1000 μm³ vs. ∼36/1000 μm³). In transgenic mini-dystrophin-expressing fibers, ultrastructure was restored (∼38/1000 μm³ counts). We suggest that in aged dystrophic toe muscle, progressive force loss is reflected by a vastly deranged micromorphology that prevents a coordinated and aligned contraction. Second harmonic generation imaging may soon be available in routine clinical diagnostics, and in this work we provide valuable imaging tools to track and quantify ultrastructural worsening in Duchenne muscular dystrophy, and to judge the beneficial effects of possible drug or gene therapies.     

The effect of the incubation period on the result of MPN enumerations of nitrite-oxidizing bacteria: theoretical considerations

Abstract A computer model based on Monod- and Haldane-kinetics was used to estimate the minimum incubation period required for MPN enumerations of nitrite-oxidizing bacteria. The minimum incubation period was defined as the time needed for one cell, present in the tubes inoculated with the highest dilutions, to grow into a population that oxidized all the nitrite present at the start of the incubation. Kinetic parameters used in the model were derived from literature data and applied in different combinations. The results show that the minimum incubation period may increase with decreasing initial nitrite concentrations in the incubation medium. They also show that the opposite trend, i.e. increasing minimum incubation periods with increasing nitrite concentration periods with increasing nitrite concentration, can be explained by introducing a term for substrate inhibition in the model. A MPN enumeration result obtained with samples from a waterlogged peat bog soil could only partly be explained by the model if only one set of parameters was used. This indicates that the community of nitrite-oxidizing bacteria in this soil is composed of at least two types of nitrite-oxidizing bacteria, with different kinetic parameters of nitrite oxidation and growth.     

Measurement of SCE frequencies in plants: a simple Feulgen-staining procedure for vicia faba

A relatively straightforward approach is described to obtain differential contrast in sister chromatids for SCE detection in Vicia faba after BrdUrd incorporation. The hydrolysis time of the well-known Feulgen reaction was extended to differentially degrade the DNA, the BrdUrd-substituted strands being more resistant. The procedure may easily be adapted for other plant species with large chromosomes.     

The shoot apical meristem restores its symplasmic organization during chilling-induced release from dormancy

The shoot apex of overwintering perennials ceases its morphogenetic activity at the end of the growing season and transforms into a bud which is dormant and freezing-tolerant. In birch (Betula pubescens) these events are triggered by short photoperiod, and involve the production of 1,3-beta-D-glucan containing sphincters on the plasmodesmata. As a result, all symplasmic pathways shut down. Here we show that breakage of bud dormancy by chilling involves restoration of the symplasmic organization of the meristem. This restoration is likely to be mediated by 1,3-beta-D-glucanase, which was present in small spherosome-like vacuoles that arose de novo during dormancy induction. During chilling these vacuoles were displaced from the bulk cytoplasm to the cortical cytoplasm where they became aligned with the plasma membrane, often associated with plasmodesmata. At this stage the enzyme also appeared outside the vacuoles. During chilling, 1,3-beta-D-glucan disappeared from the plasmodesmal channels and wall sleeves, and the plasmodesmata regained the capacity for cell-cell transport, as demonstrated by microinjection of Lucifer Yellow CH and Fluorescein-tagged gibberellic acid. Collectively, the present experiments demonstrate that restoration of the symplasmic organization of the meristem is indispensable for the release of buds from dormancy and the assumption of a proliferation-competent state, and implicate 1,3-beta-D-glucanase action at the plasmodesmata. Based on these findings we propose a model for 'dormancy cycling' which depicts the meristem as passing through three sequential states of cellular communication with characteristic sensitivities to distinct environmental cues.