Molecular diet analysis in mussels and other metazoan filter feeders and an assessment of their utility as natural eDNA samplers.

Molecular gut content analysis is a popular tool to study food web interactions and has recently been suggested as an alternative source for DNA-based biomonitoring. However, the overabundant consumer's DNA often outcompetes that of its diet during PCR. Lineage-specific primers are an efficient means to reduce consumer amplification while retaining broad specificity for dietary taxa. Here, we designed an amplicon sequencing assay to monitor the eukaryotic diet of mussels and other metazoan filter feeders and explore the utility of mussels as natural eDNA samplers to monitor planktonic communities. We designed several lineage-specific rDNA primers with broad taxonomic suitability for eukaryotes. The primers were tested using DNA extracts of different limnic and marine mussel species and the results compared to eDNA water samples collected next to the mussel colonies. In addition, we analysed several 25-year time series samples of mussels from German rivers. Our primer sets efficiently prevent the amplification of mussels and other metazoans. The recovered DNA reflects a broad dietary preference across the eukaryotic tree of life and considerable taxonomic overlap with filtered water samples. We also show the utility of a reversed version of our primers, which prevents amplification of nonmetazoan taxa from complex eukaryote community samples, by enriching fauna associated with the marine brown algae Fucus vesiculosus. Our protocol will enable large-scale dietary analysis in metazoan filter feeders, facilitate aquatic food web analysis and allow surveying of aquacultures for pathogens. Moreover, we show that mussels and other aquatic filter feeders can serve as complementary DNA source for biomonitoring.

Dispersal and adaptation strategies of the high mountain butterfly Boloria pales in the Romanian Carpathians.

BACKGROUND: Habitat quality is one main trigger for the persistence of butterflies. The effects of the influencing biotic and abiotic factors may be enhanced by the challenging conditions in high-alpine environments. To better our knowledge in this field, we performed a mark-release-recapture study with Boloria pales in the Southern Carpathians. METHODS: We analysed population structure, movement and foraging behaviour to investigate special adaptations to the alpine environment and to reveal differences between sexes. We compared these aspects in one sector with and one sector without grazing to address the effects of grazing intensity on habitat quality. RESULTS: We observed "soft" protandry, in which only a small number of males appeared before females, and an extended emergence of individuals over the observed flight period, dividing the population's age structure into three phases; both observations are considered adaptations to high mountain environments. Although both sexes were mostly sedentary, movement differences between them were obvious. Males flew larger distances than females and were more flight-active. This might explain the dimorphism in foraging behaviour: males preferred nectar sources of Asteraceae, females Caprifoliaceae. Transition from the grazed to the ungrazed sector was only observed for males and not for females, but the population density was higher and the flight distances of the individuals were significantly longer on the grazed sector compared with the ungrazed one. CONCLUSION: Soft protandry, an extended emergence of the individuals and an adapted behavioural dimorphism between sexes render to represent a good adaptation of B. pales to the harsh environmental conditions of high mountain ecosystems. However, land-use intensity apparently has severe influence on population densities and movement behaviour. To protect B. pales and other high-alpine species from the negative consequences of overgrazing, areas without or just light grazing are needed.

Assuring quality in microplastic monitoring: About the value of clean-air devices as essentials for verified data.

Avoiding aerial microfibre contamination of environmental samples is essential for reliable analyses when it comes to the detection of ubiquitous microplastics. Almost all laboratories have contamination problems which are largely unavoidable without investments in clean-air devices. Therefore, our study supplies an approach to assess background microfibre contamination of samples in the laboratory under particle-free air conditions. We tested aerial contamination of samples indoor, in a mobile laboratory, within a laboratory fume hood and on a clean bench with particles filtration during the examining process of a fish. The used clean bench reduced aerial microfibre contamination in our laboratory by 96.5%. This highlights the value of suitable clean-air devices for valid microplastic pollution data. Our results indicate, that pollution levels by microfibres have been overestimated and actual pollution levels may be many times lower. Accordingly, such clean-air devices are recommended for microplastic laboratory applications in future research work to significantly lower error rates.