Species with complex life cycles pose challenges for understanding what processes regulate population densities, especially if some life stages disperse. Most studies of such animals that are thought to be recruitment limited focus on the idea that juvenile mortality limits the density of recruits (and hence population density), fewer consider the possibility that egg supply may be important. For species that oviposit on specific substrata, environmental constraints on oviposition sites may limit egg supply. Female mayflies in the genus Baetis lay egg masses on the underside of stream rocks that emerge above the water's surface. We tested the hypothesis that egg mass densities are constrained by emergent rock densities within and between streams, by counting egg masses on emergent rocks. All emergent rocks were counted along 1-km lengths of four streams, revealing significant variation in emergent rock density within streams and a more than three-fold difference between streams. In each stream, egg mass density increased with the density of emergent rocks in 30-m stretches. We used regression equations describing these small-scale relationships, coupled with the large-scale spatial variation of emergent rocks, to estimate egg mass densities for each 1-km stream length, a scale relevant to population processes. Scaled estimates were positively associated with emergent rock density and provided better estimates than methods that ignored environmental variation. Egg mass crowding was inversely related to emergent rock density at the stream scale, a pattern consistent with the idea that oviposition substrata were in short supply in streams with few emergent rocks, but crowding did not compensate entirely for differences in emergent rock densities. The notion that egg supply, not larval mortality, may limit population density is an unusual perspective for stream insects. Environmental constraints on egg supply may be widespread among other species with specialised oviposition behaviours.

Excessive sediment pressure on aquatic habitats is of global concern. A unique dataset, comprising instantaneous measurements of deposited fine sediment in 230 agricultural streams across England and Wales, was analysed in relation to 20 potential explanatory catchment and channel variables. The most effective explanatory variable for the amount of deposited sediment was found to be stream power, calculated for bankfull flow and used to index the capacity of the stream to transport sediment. Both stream power and velocity category were highly significant (p ≪ 0.001), explaining some 57% variation in total fine sediment mass. Modelled sediment pressure, predominantly from agriculture, was marginally significant (p<0.05) and explained a further 1% variation. The relationship was slightly stronger for erosional zones, providing 62% explanation overall. In the case of the deposited surface drape, stream power was again found to be the most effective explanatory variable (p<0.001) but velocity category, baseflow index and modelled sediment pressure were all significant (p<0.01); each provided an additional 2% explanation to an overall 50%. It is suggested that, in general, the study sites were transport-limited and the majority of stream beds were saturated by fine sediment. For sites below saturation, the upper envelope of measured fine sediment mass increased with modelled sediment pressure. The practical implications of these findings are that (i) targets for fine sediment loads need to take into account the ability of streams to transport/retain fine sediment, and (ii) where agricultural mitigation measures are implemented to reduce delivery of sediment, river management to mobilise/remove fines may also be needed in order to effect an improvement in ecological status in cases where streams are already saturated with fines and unlikely to self-cleanse.

1. Detrimental impacts of excessive fine-grained sediment inputs to streams and rivers are well established. What is less well understood is the susceptibility of different elements of the freshwater biota to such perturbations and how such knowledge of their susceptibility could aid in identifying where excessive fine-grained sediment is impairing ecological condition. 2. Following the collection of biological and sediment data from 179 streams across England and Wales, representative of a range of river types over a gradient of fine sediment loading, objective statistical approaches were applied to establish relationships between the macroinvertebrate assemblage and fine-grained sediment inputs to river channels. 3. Having factored out that portion of the biological variation associated with natural environmental gradients, a model comprising mass of organic sediment in erosional areas of the stream bed [predominantly associated with the first axis of the partial canonical correspondence analysis (pCCA)], and mass of fine-grained sediment in the surface drape of depositional areas and % organic content in erosional areas (associated with the second axis of the pCCA) as explanatory variables best accounted for the residual variation in the macroinvertebrate assemblage. 4. The relative position of taxa along both axes of the pCCA, provided a ranking of taxa in relation to the two gradients of fine-grained sediment and provided the basis for a new empirically derived diagnostic index for fine-grained sediment stress in rivers. Two sub-indices were derived to capture the assemblage responses to both the gradient of organic sediment in erosional areas and the gradient of total fines in depositional areas. The two sub-indices were then combined to derive the new combined fine sediment index (CoFSI sp ). 5. The index was tested on an independent test data set (comprising 127 samples from 83 sites) and was found to provide a robust indication of benthic fine-grained sediment conditions (Spearman's rank correlations q = À0.519 to À0.703). The strength of correlation with the total fine-grained sediment gradient was always greater than that for other routinely used indices, confirming that CoFSI sp offered additional explanatory power when assessing this stressor of aquatic environments.

Over the last decade, steady advancements have been made in the use of DNA-based methods for detection of species in a wide range of ecosystems. This progress has culminated in molecular monitoring methods being employed for the detection of several species for enforceable management purposes of endangered, invasive, and illegally harvested species worldwide. However, the routine application of DNA-based methods to monitor whole communities (typically a metabarcoding approach) in order to assess the status of ecosystems continues to be limited. In aquatic ecosystems, the limited use is particularly true for macroinvertebrate communities. As part of the DNAqua-Net consortium, a structured discussion was initiated with the aim to identify potential molecular methods for freshwater macroinvertebrate community assessment and identify important knowledge gaps for their routine et al.: Biomonitoring macroinvertebrates with molecular methods 50 application. We focus on three complementary DNA sources that can be metabarcoded: 1) DNA from homogenised samples (bulk DNA), 2) DNA extracted from sample preservative (fixative DNA), and 3) environmental DNA (eDNA) from water or sediment. We provide a brief overview of metabarcoding macroinvertebrate communities from each DNA source and identify challenges for their application to routine monitoring. To advance the utilisation of DNA-based monitoring for macroinvertebrates, we propose an experimental design template for a series of methodological calibration tests. The template compares sources of DNA with the goal of identifying the effects of molecular processing steps on precision and accuracy. Furthermore, the same samples will be morphologically analysed, which will enable the benchmarking of molecular to traditional processing approaches. In doing so we hope to highlight pathways for the development of DNA-based methods for the monitoring of freshwater macroinvertebrates.

Summary1. Predicting population dynamics at large spatial scales requires integrating information about spatial distribution patterns, inter-patch movement rates and within-patch processes. Advective dispersal of aquatic species by water movement is considered paramount to understanding their population dynamics. Rivers are model advective systems, and the larvae of baetid mayflies are considered quintessential dispersers. Egg laying of baetids along channels is patchy and reflects the distribution of oviposition sites, but larvae are assumed to drift frequently and far, thereby erasing patterns created during oviposition. Dispersal kernels are often overestimated, however, and empirical tests of such assumptions are warranted because of the pivotal role distribution patterns can have on populations. 2. We tested empirically whether the egg distribution patterns arising from oviposition behaviours persisted and were reflected in the distribution patterns of larval Baetis rhodani. In field surveys, we tested for associations between egg mass and larval densities over 1 km lengths of four streams. A control species, the mayfly Ephemerella ignita, was employed to test for covarying environmental factors. We estimated drift rates directly to test whether larvae dispersed between riffles (patches of high egg mass density) and whether drift rates were density-dependent or density-relatedexpected outcomes if drift erases patterns established by maternal behaviours. 3. Positive associations between egg masses and larval benthic densities were found for neonate and mid-stage larvae of Baetis, but not the control species, suggesting persistence of the patchy distribution patterns established at oviposition. Drift rates were high, and riffles were net exporters of neonate and mid-stage larvae, but drift rates were unrelated to benthic densities and few drifters reached the next riffle. Riffles were sinks for large larvae, suggesting ontogenetic shifts in habitat use, but little long-distance dispersal. 4. Overall, the results suggest that most neonate and mid-stage larvae of B. rhodani remain close to the natal riffle, and late-stage larvae disperse shorter distances than routinely assumed. The persistence of maternal effects on distribution patterns well into juvenile life of an allegedly iconic disperser suggests that traditional models of how dispersal influences the population dynamics of many lotic invertebrates may be incorrect.