Particle size influences decay rates of environmental DNA in aquatic systems

Environmental DNA (eDNA) analysis is a powerful tool for remote detection of target organisms. However, obtaining quantitative and longitudinal information from eDNA data is challenging, requiring a deep understanding of eDNA ecology. Notably, if the various size components of eDNA decay at different rates, and we can separate them within a sample, their changing proportions could be used to obtain longitudinal dynamics information on targets. To test this possibility, we conducted an aquatic mesocosm experiment in which we separated fish-derived eDNA components using sequential filtration to evaluate the decay rate and changing proportion of various eDNA particle sizes over time. We then fit four alternative mathematical decay models to the data, building towards a predictive framework to interpret eDNA data from various particle sizes. We found that medium-sized particles (1–10 μm) decayed more slowly than other size classes (i.e., <1 and > 10 μm), and thus made up an increasing proportion of eDNA particles over time. We also observed distinct eDNA particle size distribution (PSD) between our Common carp and Rainbow trout samples, suggesting that target-specific assays are required to determine starting eDNA PSDs. Additionally, we found evidence that different sizes of eDNA particles do not decay independently, with particle size conversion replenishing smaller particles over time. Nonetheless, a parsimonious mathematical model where particle sizes decay independently best explained the data. Given these results, we suggest a framework to discern target distance and abundance with eDNA data by applying sequential filtration, which theoretically has both metabarcoding and single-target applications.     

OBSERVATIONS ON SPERM PENETRATION IN THE RAT

The structural aspects of sperm penetration in the rat egg were investigated by electron microscopy. Eggs were recovered at intervals between 8 and 10:30 A.M. from females which had mated during the previous night. The oviducts were flushed with hyaluronidase and the eggs transferred into a 2 per cent osmium tetroxide solution, buffered at pH 7.8. After fixation, the eggs were mounted individually in agar, dehydrated in ethyl alcohol, and embedded in butyl-methyl methacrylate (3:1). The sperm penetrating the egg is covered by a plasma membrane which is present only on the side facing toward the zona pellucida; no membrane is visible on the side facing toward the vitellus. The sperm plasma membrane becomes continuous with the egg plasma membrane and forms a deep fold around the entering sperm. Cross-sections through the sperm midpiece in the perivitelline space show an intact plasma membrane. At the place of entrance, the plasma membrane of the sperm appears to fuse with the egg plasma membrane. After the sperm has penetrated the vitellus, it has no plasma membrane at all. The nuclear membrane is also absent. These observations suggest a new hypothesis for sperm penetration. After the sperm has come to lie on the plasma membrane of the egg, the egg and sperm plasma membranes rupture and then fuse with one another to form a continuous cell membrane over the egg and the outer surface of the sperm. As a result the sperm comes to lie inside the vitellus, leaving its own plasma membrane incorporated into the egg membrane at the surface of the egg.     

THE DIRECT ISOLATION OF THE MITOTIC APPARATUS

A method for isolating the mitotic apparatus from dividing sea urchin eggs without the use of ethyl alcohol or of detergents is described. In the present method, the eggs are dispersed directly in a medium containing 1 M (to 1.15 M) sucrose, 0.15 M dithiodiglycol, and 0.001 M Versene at pH 6, releasing the visibly intact mitotic apparatus. The method is designed for studies of enzyme activities, lipid components, and the variables affecting the stability of the apparatus.     

RESPONSE OF LEAF ANATOMY AND PHOTOSYNTHETIC CAPACITY IN ALOCASIA MACRORRHIZA (ARACEAE) TO A TRANSFER FROM LOW TO HIGH LIGHT

Alocasia macrorrhiza plants were grown in 1% and 20% full sunlight, and their leaf anatomical and physiological parameters were measured. Total leaf thickness was 41% greater and mesophyll thickness was 52% greater in high-light leaves than in low-light leaves. This increase in thickness resulted from both increased cell size and number. Maximum leaf photosynthetic capacity was also 66% greater in high- than in low-light leaves. When low-light plants were transferred to high light, the thickness of mature leaves did not increase but the thickness of the first leaf to expand after the transfer was significantly greater than that of the low-light leaves. Thus, only leaves that were still expanding at the time of transfer developed leaf thickness greater than plants remaining in low light. Fully mature leaves showed no change in photosynthetic capacity in response to transfer. Leaves that had just completed expansion at the time of low- to high-light transfer were able to develop slightly higher maximum photosynthetic capacities than older leaves. However, full photosynthetic acclimation to the new light environment did not occur until the second new leaf expanded after transfer. These results are discussed in relation to the timing and mechanisms of whole plant acclimation to increased light.     

CALLING SONG DISPLACEMENT IN A ZONE OF OVERLAP AND HYBRIDIZATION

The ground crickets Allonemobius fasciatus and A. socius meet in a mosaic zone of overlap and hybridization stretching from the East Coast to at least Illinois. To test whether male calling song differences were enhanced in sympatry, we analyzed the songs of crickets from inside and outside the zone of overlap along two transects. No evidence of calling song displacement was found in A. socius males from populations within the zone of overlap. On the other hand, A. fasciatus displayed calling song displacement in three populations. Our results are consistent with the hypothesis that the selective pressure exerted by the challenge from a related species is frequency dependent. While not a conclusive demonstration, the observed shifts in calling song are strongly suggestive of reproductive character displacement.     

SENESCENCE IN NATURAL POPULATIONS OF MAMMALS: A COMPARATIVE STUDY

Here, I use published mortality data from 56 natural populations of mammals to examine evidence for senescence, an increase in the probability of mortality with age. Data on extent of senescence and life history characteristics are compared across taxa in an attempt to test theories for the evolution of senescence in natural populations. In accord with theoretical expectation, senescence is highest in short-lived species with short generation times. In contrast to theoretical expectation, however, senescent increases in mortality rate do not begin until well after age at maturity in most cases. I also present evidence in support of the hypothesis that senescence will be lower in large-brained taxa.     

SYSTEMATIC ANALYSIS OF SPHAEROPLEA (CHLOROPHYCEAE)1

A systematic investigation of the genus Sphaeroplea was conducted using cladistic analyses of both structural and isozyme characters for the same set of taxa. The structural data were not able to fully resolve some of the taxa while the isozyme data did produce a tree in which all nodes were supported by data. The structural characters were relatively consistent with one another, whereas the isozyme characters were much less internally consistent. Results from independent, cladistic analyses of both data sets support the concept that among those Sphaeroplea species investigated, S. fragilis Buchheim et Hoffman had an early divergence. The two data sets differed primarily in that the structural data support monophyly of the genus Sphaeroplea and the isozyme data do not. The greater relative consistency of the structural data suggests better support for trees inferred from its analysis. Furthermore, searches for character congruence between the two data sets revealed isozyme data which support monophyly of the genus Sphaeroplea, but had been overwhelmed by conflicting isozyme characters.