Chitinase digestion for the analysis of microplastics in chitinaceous organisms using the terrestrial isopod Oniscus asellus L. as a model organism

Chitinaceous organisms have been found to ingest microplastic; however, a standardised, validated, and time- and cost-efficient method for dissolving these organisms without affecting microplastic particles is still required. This study tested four protocols for dissolving organisms with a chitin exoskeleton: 1) potassium hydroxide (KOH) + chitinase, 2) Creon® + chitinase, 3) hydrogen peroxide (H2O2) + chitinase, and, 4) Nitric Acid (HNO3) + hydrogen peroxide (H2O2). The effects on microplastics composed of eight different polymers were also tested. The use of H2O2 followed by chitinase was found to be a highly efficient method. The three other protocols either did not digest the chitin sufficiently or negatively affected the tested polymers. A recovery test using microplastic fibres, beads and tyre particles revealed high recovery rates of 0.85, 0.89 and 1 respectively. This further supported the applicability of the H2O2 and chitinase (protocol 3) for dissolving chitinaceous organisms. Thus, we recommend that future investigations of microplastic (0.05 μm–5000 μm) in chitinaceous organisms (0.3 cm–5 cm) utilise the here presented methodology. This represents an important component of the ongoing validation and harmonization of methodological approaches that are urgently needed for the advancement of microplastic assessments globally.publishedVersio

Local and landscape scale determinants of biodiversity and conservation value of macroinvertebrate assemblages in ponds across an urban land-use gradient

Urbanisation represents a growing threat to natural communities across the globe. Small aquatic habitats such as ponds are especially vulnerable and are often poorly protected by legislation. Many ponds are threatened by development and pollution from the surrounding landscape, yet their biodiversity and conservation value remain poorly described. Here we report the results of a survey of 30 ponds along an urban land-use gradient in the West Midlands, UK. We outline the environmental conditions of these urban ponds to identify which local and landscape scale environmental variables determine the biodiversity and conservation value of the macroinvertebrate assemblages in the ponds. Cluster analysis identified four groups of ponds with contrasting macroinvertebrate assemblages reflecting differences in macrophyte cover, nutrient status, riparian shading, the nature of the pond edge, surrounding land-use and the availability of other wetland habitats. Pond conservation status varied markedly across the sites. The richest macroinvertebrate assemblages with high conservation value were found in ponds with complex macrophyte stands and floating vegetation with low nutrient concentrations and little surrounding urban land. The most impoverished assemblages were found in highly urban ponds with hard-engineered edges, heavy shading and nutrient rich waters. A random forest classification model revealed that local factors usually had primacy over landscape scale factors in determining pond conservation value, and constitute a priority focus for management

High-throughput analyses and Bayesian network modeling highlight novel epigenetic Adverse Outcome Pathway networks of DNA methyltransferase inhibitor mediated transgenerational effects

A number of epigenetic modulating chemicals are known to affect multiple generations of a population from a single ancestral exposure, thus posing transgenerational hazards. The present study aimed to establish a high-throughput (HT) analytical workflow for cost-efficient concentration-response analysis of epigenetic and phenotypic effects, and to support the development of novel Adverse Outcome Pathway (AOP) networks for DNA methyltransferase (DNMT) inhibitor-mediated transgenerational effects on aquatic organisms. The model DNMT inhibitor 5-azacytidine (5AC) and the model freshwater crustacean Daphnia magna were used to generate new experimental data and served as prototypes to construct AOPs for aquatic organisms. Targeted HT bioassays (DNMT ELISA, MS-HRM and qPCR) in combination with multigenerational ecotoxicity tests revealed concentration-dependent transgenerational (F0-F3) effects of 5AC on total DNMT activity, DNA promoter methylation, gene body methylation, gene transcription and reproduction. Top sensitive toxicity pathways related to 5AC exposure, such as apoptosis and DNA damage responses were identified in both F0 and F3 using Gaussian Bayesian network modeling. Two novel epigenetic AOP networks on DNMT inhibitor mediated one-generational and transgenerational effects were developed for aquatic organisms and assessed for the weight of evidence. The new HT analytical workflow and AOPs can facilitate future ecological hazard assessment of epigenetic modulating chemicals

Plastic recycling plant as a point source of microplastics to sediment and macroinvertebrates in a remote stream

Microplastic is now ubiquitous in freshwater, sediment and biota, globally. This is as a consequence of inputs from, for example, waste mismanagement, effluents from wastewater treatment plants and surface runoff from agricultural areas. In this study, we investigated point source pollution of plastic to an upland stream, originating from a recycling plant that recycles polyethylene film in a remote area of Norway. Sediment (~2 kg) and macroinvertebrates (549 individuals in total) were sampled at one site upstream and two sites downstream of the recycling plant to study microplastic deposition and food web uptake. In total, 340 microplastic films were identified through a combination of visual and µFTIR analysis in the sediment samples. This corresponded to a concentration of 0.23 (± 0.057) items per g sediment upstream of the plastic recycling plant and 0.45 (± 0.017) and 0.58 (± 0.34) items per g downstream. The dominant plastic polymer was polyethylene, which increased significantly downstream of the plastic recycling plant. This indicates the role of the plastic recycling plant as a point source for microplastic in this catchment. Among the three sites investigated, a fairly constant concentration of polypropylene was found, indicating a diffuse source of polypropylene films across the catchment possibly relating to low-intensity agricultural land-use. Low levels of polyethylene were also observed upstream, which may be linked to either local or longer-distance atmospheric transport. Despite the considerable presence of microplastic in sediments, concentrations in macroinvertebrates were extremely low with only a single microplastic particle identified in the total of 549 macroinvertebrates—belonging to three different feeding groups—investigated. Our study suggests that: 1) microplastic pollution can be transferred to remote areas as unintended losses from recycling facilities, 2) remote areas with limited land-use pressure still have detectable levels of microplastic and 3) microplastic is only taken up by stream macroinvertebrates to a limited degree despite relatively high sediment concentrations, and thus there are no strong indications for ecological risks posed by microplastic to this ecological group at this location.publishedVersio

A global perspective on the application of riverine macroinvertebrates as biological indicators in Africa, South-Central America, Mexico and Southern Asia

The aim of this study is to generate a first global overview of pressures and methods used to assess the environmental quality of rivers and streams using macroinvertebrates. In total, 314 peer-review studies were reviewed, published in the period 1997 – 2018, from developing economies in Africa, South-Central America, Mexico and Southern Asia. To establish a global perspective, the results from the literature review were compared to other compiled datasets, biomonitoring manuals, environmental surveys and literature reviews from Europe, North America and Australasia. The literature review from the developing economies showed that sampling was most usual during baseflow, using kick- or Surber sampling, with taxonomical identification levels mostly to genus or family. Assessments were most often done using metrics (singular and multimetrics; > 70% of the applications) and were based on community attributes related to richness and dominance (58% of studies), sensitivity (40%), diversity by heterogeneity (32%) and functional traits (25%). Within each category, the most used metrics were the richness and dominance of Ephemeroptera, Plecoptera and Trichoptera (EPT), Biological Monitoring Working Party scoring systems (BMWP/ASPT), Shannon-Wiener diversity and feeding traits. Overall, 92% of the reviewed studies reported that the use of macroinvertebrates, at least in some of their responses, was successful in detecting degradation of environmental quality in the investigated rivers. Given the many similarities in applied methods worldwide, at present, we consider that a global assessment of riverine environmental quality can be feasible by using family level identifications of macroinvertebrate samples. We propose a global common metric (multimetric), comprising three of the most common river assessment metrics from the reviewed literature, but also elsewhere, namely the BMWP/ASPT, Shannon-Wiener diversity and richness of EPT. Recent concerns regarding the global state of nature and consequences for freshwater communities, as reported by the intergovernmental science-policy platform on biodiversity and ecosystem services (IPBES), emphasize the urgent need for such a synthesis

Re-Meandering of Lowland Streams: Will Disobeying the Laws of Geomorphology Have Ecological Consequences?

Abstract We evaluated the restoration of physical habitats and its influence on macroinvertebrate community structure in 18 Danish lowland streams comprising six restored streams, six streams with little physical alteration and six channelized streams. We hypothesized that physical habitats and macroinvertebrate communities of restored streams would resemble those of natural streams, while those of the channelized streams would differ from both restored and near-natural streams. Physical habitats were surveyed for substrate composition, depth, width and current velocity. Macroinvertebrates were sampled along 100 m reaches in each stream, in edge habitats and in riffle/run habitats located in the center of the stream. Restoration significantly altered the physical conditions and affected the interactions between stream habitat heterogeneity and macroinvertebrate diversity. The substrate in the restored streams was dominated by pebble, whereas the substrate in the channelized and natural streams was dominated by sand. In the natural streams a relationship was identified between slope and pebble/gravel coverage, indicating a coupling of energy and substrate characteristics. Such a relationship did not occur in the channelized or in the restored streams where placement of large amounts of pebble/gravel distorted the natural relationship. The analyses revealed, a direct link between substrate heterogeneity and macroinvertebrate diversity in the natural streams. A similar relationship was not found in either the channelized or the restored streams, which we attribute to a de-coupling of the natural relationship between benthic community diversity and physical habitat diversity. Our study results suggest that restoration schemes should aim at restoring the natural physical structural complexity in the streams and at the same time enhance the possibility of re-generating the natural geomorphological processes sustaining the habitats in streams and rivers. Documentation of restoration efforts should be intensified with continuous monitoring of geomorphological and ecological changes including surveys of reference river systems