Automated identification of aquatic insects: A case study using deep learning and computer vision techniques.

Despite huge attention in other domains, deep learning is only very slowly beginning to be applied in biodiversity research. Mayflies (Ephemeroptera), stoneflies (Plecoptera) and caddisflies (Trichoptera), often abbreviated as EPT, are frequently used for freshwater biomonitoring due to their large numbers and sensitivity to environmental changes. However, the unambiguous morphological identification of EPT species is a challenging, but fundamental task. Morphological identification of these freshwater insects is therefore not only extremely time-consuming and costly, but also often leads to misjudgments or generates datasets with low taxonomic resolution. Here, we investigated the application of deep learning to increase the efficiency and taxonomic resolution of biomonitoring programs. Our database contains 90 EPT taxa (genus or species level), with the number of images per category ranging from 21 to 300 (16,650 in total). Upon completion of training, a CNN (Convolutional Neural Network) model was created, capable of automatically classifying these taxa into their appropriate taxonomic categories with an accuracy of 98.7 %. For the extensive set of 68 tested taxa, our model achieved a perfect classification rate of 100 %. We achieved noteworthy classification accuracy with morphologically closely related taxa within the training data (e.g., species of the genus Baetis, Hydropsyche, Perla). Gradient-weighted Class Activation Mapping (Grad-CAM) visualized the morphological features responsible for the classification of the treated species in the CNN models. In the Ephemeroptera, the head was the most important feature, while the thorax and abdomen were equally important for the classification of Plecoptera taxa. For the order Trichoptera, the head and thorax were almost equally important. Our database is recognized as the most extensive aquatic insect database, notably distinguished by its wealth of included categories (taxa). Our approach can help solve long-standing challenges in biodiversity research and address pressing issues in monitoring programs by saving time in sample and data processing.

Beneath the surface: Decoding the impact of Chironomus riparius bioturbation on microplastic dispersion in sedimentary matrix.

A detailed understanding of microplastics (MPs) behaviour in freshwater ecosystems is crucial for a proper ecological assessment. This includes the identification of significant transport pathways and net accumulation zones, considering their inherent, and already proven influence on aquatic ecosystems. Bioavailability of toxic agents is significantly influenced by macroinvertebrates' behaviour, such as bioturbation and burrowing, and their prior exposure history. This study investigates the effect of bioturbation activity of Chironomus riparius Meigen, 1804 on the vertical transfer of polyethylene MPs ex-situ. The experimental setup exposes larvae to a scenario of 10× the environmentally relevant high concentration of MPs (80 g m-2). Bioturbation activity was estimated using sediment profile imaging with luminophore tracers. This study demonstrated that spherical MPs are vertically transferred in the sediment due to the bioturbation activity of C. riparius larvae and that their presence influences the intensity of the bioturbation activity over time. The present findings provide a noteworthy contribution to the understanding of the relationship between ecosystem engineers and the dispersion and accumulation of MPs within freshwater ecosystems.

The influence of spatial processes on fish community structure: using a metacommunity framework for freshwater bioassessment.

The use of biological indicators in a bioassessment approach is important for inferences of anthropogenic stress in routine monitoring programs. One of the primary assumptions of bioassessment is that biological indicators observed at specific sampling sites will allow for inferences of local environmental quality; however, this assumption requires a reliable understanding of dispersal processes, which is particularly relevant in river systems due to their dendritic network. Inter-stream dispersal between different points of the river network could bias bioassessment, especially for highly mobile organisms like fish. Here, we examine standard biological metrics used in routine biomonitoring to determine how spatial variables, including dispersal, influence inferences between fish populations and environmental gradients. Using redundancy analysis (RDA) and variation partitioning, we tested the relative influence of both environmental and spatial variables on fish community structure and related community metrics. Fish were collected from 99 sampling sites distributed across 44 rivers and streams of the Great Morava River Basin, Serbia. Electroconductivity, the percentage of agricultural areas, dissolved oxygen, ammonia, and nitrate-nitrogen were found to be significant environmental variables, while ten spatial predictors from broad- to small-scales were found to influence fish community structure and community metrics. Our results suggest that contemporary dispersal among streams influences fish community composition, but that trait-based metrics are less sensitive than basic measures of diversity to spatial processes. Our results highlight the need for spatially independent sampling, as well as the need to consider dispersal-based processes in routine biomonitoring.

Toxic effects of a cyanobacterial strain on Chironomus riparius larvae in a multistress environment.

Cyanobacteria and their toxic metabolites present a global threat to water habitats, but their impact on aquatic organisms in a multistress environment has been poorly investigated. Here we present the results of a survey on the effects of the toxic cyanobacterial strain Trichormus variabilis (heterotypic synonym Anabaena variabilis), and its toxic metabolite, cyanotoxin microcystin-LR, on Chironomus riparius larvae in a multistress environment. An environmentally relevant concentration of microcystin-LR (0.01 mg/L) caused an increase in larvae mortality in an acute toxicity test, which became greater in the presence of environmental stressors (NO3-, NH4+, PO43- and Cd2+), pointing to an additive effect of these agents. Chronic exposure of C. riparius larvae to the microcystin-LR producing strain of T. variabilis in a multistress environment led to a reduction in the larval mass and hemoglobin concentration, and it induced DNA damage in larval somatic cells. The results revealed the additive effect of microcystin-LR in combination with all three tested stressors (NO3-, NH4+, PO43-), and the deleterious effect of chronic exposure of C. riparius larvae to the microcystin-LR producing T. variabilis in a multistress environment. However, the present study further emphasizes the importance of investigating interactions between stressors and cyanotoxins, and their effect on aquatic organisms.

Effects of a microplastic mixture differ across trophic levels and taxa in a freshwater food web: In situ mesocosm experiment.

The ubiquitous presence of microplastics (MP) in aquatic ecosystems can affect organisms and communities in multiple ways. While MP research on aquatic organisms has primarily focused on marine ecosystems and laboratory experiments, the community-level effects of MP in freshwaters, especially in lakes, are poorly understood. To examine the impact of MP on freshwater lake ecosystems, we conducted the first in situ community-level mesocosm experiment testing the effects of MP on a model food web with zooplankton as main herbivores, odonate larvae as predators, and chironomid larvae as detritivores for seven weeks. The mesocosms were exposed to a mixture of the most abundant MP polymers found in freshwaters, added at two different concentrations in a single pulse to the water surface, water column and sediment. Water column MP concentrations declined sharply during the first two weeks of the experiment. Contrary to expectations, MP ingestion by zooplankton was low and limited mainly to large-bodied Daphnia, causing a decrease in biomass. Biomass of the other zooplankton taxa did not decrease. Presence of MP in the faecal pellets of odonate larvae that fed on zooplankton was indicative of a trophic transfer of MP. The results demonstrated that MP ingestion varies predictably with MP size, as well as body size and feeding preference of the organism, which can be used to predict the rates of transfer and further effects of MP on freshwater food webs. For chironomids, MP had only a low, short-term impact on emergence patterns while their wing morphology was significantly changed. Overall, the impact of MP exposure on the experimental food web and cross-ecosystem biomass transfer was lower than expected, but the experiment provided the first in situ observation of MP transfer to terrestrial ecosystems by emerging chironomids.

Implications of local niche- and dispersal-based factors that may influence chironomid assemblages in bioassessment.

Local environmental factors and dispersal-based processes can both influence the structure of metacommunities in freshwater ecosystems. Describing these patterns is especially important for biomonitoring approaches that are based on inferences made from benthic macroinvertebrate assemblages. Here, we examine the metacommunity structure of chironomid assemblages collected from 28 sampling stations along the Southern Morava River, Serbia. We examined the extent of dispersal-based processes along a temporal scale. We obtained 8 models for the different sampling seasons that determined the spatial variables that best explained variability in chironomid assemblages. Spatial processes were found to be a significant predictor of variation for chironomids during the late winter/spring (March and May) and autumn (October and November), concordant with the known phenology of common taxa. Species sorting and mass effects were found to be significant processes that structured the chironomid metacommunity. In addition, biological interactions, inferred from fish biomass, and habitat traits, demonstrated by macrophyte and riparian vegetation, were found to influence species sorting. A high variability of chironomid metacommunity structure across sampling seasons suggests that monitoring programs that include macroinvertebrates in bioassessment should avoid months with pronounced spatial processes, and consequently maximize a correlation between community structure and local environmental factors.

Taxon-specific sensitivities to flow intermittence reveal macroinvertebrates as potential bioindicators of intermittent rivers and streams.

As complex mosaics of lotic, lentic, and terrestrial habitats, intermittent rivers and ephemeral streams (IRES) support high biodiversity. Despite their ecological importance, IRES are poorly represented in routine monitoring programs, but recent recognition of their considerable-and increasing-spatiotemporal extent is motivating efforts to better represent IRES in ecological status assessments. We examine response patterns of aquatic macroinvertebrate communities and taxa to flow intermittence (FI) across three European climatic regions. We used self-organizing map (SOM) to ordinate and classify sampling sites based on community structure in regions with continental, Mediterranean and oceanic climates. The SOM passively introduced FI, quantified as the mean annual % flow, and visualized its variability across classified communities, revealing a clear association between community structure and FI in all regions. Indicator species analysis identified taxa indicative of low, intermediate and high FI. In the continental region, the amphipod Niphargus was indicative of high FI and was associated with groundwater-fed IRES, whereas indicators of Mediterranean IRES comprised Odonata, Coleoptera and Heteroptera taxa, which favor lentic conditions. In the oceanic region, taxa indicative of relatively high FI included leuctrid stoneflies and a limnephilid caddisfly, likely reflecting the colonization of IRES by aerial adults from nearby perennial reaches. The Diptera families Chironomidae and Simuliidae showed contrasting FI preferences among regions, reflecting environmental heterogeneity between regions and the coarse taxonomic resolution to which these organisms were identified. These region-specific community and taxon responses of aquatic biota to FI highlight the need to adapt standard biotic indices to enable effective ecological status assessments in IRES.

In Situ Effects of a Microplastic Mixture on the Community Structure of Benthic Macroinvertebrates in a Freshwater Pond.

Benthic communities contain some of the most threatened organisms in aquatic habitats due to different anthropogenic pressures. The high abundance of microplastics in sediments will continue to increase in the future, further increasing the probability of interactions between macroinvertebrates and microplastics. In the present study, a benthic community in a relatively pristine shallow pond was exposed either to an environmentally relevant high concentration of a microplastic mixture of 80 g m-2 in the sediment, or a control sediment, without the addition of microplastics. The mixture of microplastics contained irregularly shaped polyethylene, polyvinyl chloride, and polyamide in a ratio of 50:25:25%, respectively. The in situ experiment lasted for 100 d. The total number of taxa that colonized the microcosms was 22 (17 in the control and 18 in the microplastic treatment), and the colonization was not affected by the treatment. The most dominant group within the macroinvertebrate community was the dipteran family Chironomidae, in both the control and the microplastic treatment. No significant differences in the abundance and biomass at a community level were recorded between the groups by permutational multivariate analysis of variance (F = 0.993, p = 0.456 and F = 0.344, p = 0.797, respectively). The mixture of microplastics did not influence the abundance or biomass of the functional feeding groups (F = 1.810, p = 0.137 and F = 0.377, p = 0.736, respectively). The species richness, species abundance, species biomass, Shannon's diversity index, and Simpson's index of diversity showed no statistically significant differences between the control and treatment groups. Czekanowski's quantitative similarity index indicated that 84% of the community remained unaffected after microplastic exposure. Environ Toxicol Chem 2022;41:888-895. © 2021 SETAC.

The application of Uniform Manifold Approximation and Projection (UMAP) for unconstrained ordination and classification of biological indicators in aquatic ecology.

The analysis of community structure in studies of freshwater ecology often requires the application of dimensionality reduction to process multivariate data. A high number of dimensions (number of taxa/environmental parameters × number of samples), nonlinear relationships, outliers, and high variability usually hinder the visualization and interpretation of multivariate datasets. Here, we proposed a new statistical design using Uniform Manifold Approximation and Projection (UMAP), and community partitioning using Louvain algorithms, to ordinate and classify the structure of aquatic biota in two-dimensional space. We present this approach with a demonstration of five previously published datasets for diatoms, macrophytes, chironomids (larval and subfossil), and fish. Principal Component Analysis (PCA) and Ward's clustering were also used to assess the comparability of the UMAP approach compared to traditional approaches for ordination and classification. The ordination of sampling sites in 2-dimensional space showed a much denser, and easier to interpret, grouping using the UMAP approach in comparison to PCA. The classification of community structure using the Louvain algorithm in UMAP ordinal space showed a high classification strength for data with a high number of dimensions than the cluster patterns obtained with the use of a Ward's algorithm in PCA. Environmental gradients, presented via heat maps, were overlayed with the ordination patterns of aquatic communities, confirming that the ordinations obtained by UMAP were ecologically meaningful. This is the first study that has applied a UMAP approach with classification using Louvain algorithms on ecological datasets. We show that the performance of local and global structures, as well as the number of clusters determined by the algorithm, make this approach more powerful than traditional approaches.

Bioassessment of the ecological integrity of freshwater ecosystems using aquatic macroinvertebrates: the case of Sable Island National Park Reserve, Canada.

Due to ubiquitous distribution of taxa, relatively low-cost and efficient sampling procedure, and known responses to environmental gradients, macroinvertebrate indicators are often a central component of biological monitoring of freshwater resources. This study examined establishing a baseline reference of benthic macroinvertebrate indicators in a biomonitoring approach as a means for monitoring the freshwater ponds of Sable Island National Park Reserve (SINPR), Canada. We compared water quality parameters monitored from 2015 to 2019 to a biomonitoring approach deployed in May, June, and August of 2019. A total of 27 taxa were recorded from the 30,226 specimens collected, with highest abundances of Corixidae, Amphipoda, Oligochaeta, and chironomid species Polypedilum bicrenatum. We found significant variability of community structure between different months of sampling (p = 0.001) and between ponds (p < 0.0001). A high correlation was found between dissolved organic carbon, sulfate, and the diversity of macroinvertebrate indicators, while conductivity, ammonia, and calcium were found to be correlated with species richness. While we found that water chemistry parameters exhibited spatial and temporal differences, the diversity of macroinvertebrate indicators is likely to be a more resilient metric for comparison between ponds. Further, our findings demonstrate that biomonitoring can be effective in systems with a low number of small, shallow, freshwater pond ecosystems. As our study deployed a high-resolution identification of biological indicators, we were able to establish a baseline reference for future monitoring as well as identify specific associations between pond water quality and biological assemblages that can be used as a context for the management of SINPR's freshwater resources. Continued monitoring of these ecosystems in future years will help to understand long-term environmental changes on the island.

Chironomidae (Diptera) of Croatia with notes on the diversity and distribution in various habitat types.

The family of non-biting midges (Diptera, Chironomidae) represent one of the most common and abundant group in freshwaters, inhabiting a wide variety of habitats and ecological niches. Although there is a long tradition of limnological research in Croatia, to date no extensive species list has been made for this insect family. Thus, we summarized the results of our research and reviewed published data on chironomid fauna of Croatia in various freshwater habitat types, including extreme ones, such as caves, peatbogs or marine littoral. A total of 239 species were recorded representing five subfamilies: Chironominae (125 species) Orthocladiinae (83 species), Tanypodinae (23 species), Diamesinae (6 species) and Prodiamesinae (2 species). The most frequent species were Cricotopus bicinctus, Dicrotendipes nervosus, Synorthocladius semivirens, each found in more than 20% of the studied localities. Ablabesmyia monilis and Procladius choreus represented the most frequent Tanypodinae species, recorded in more than 12% and 17% of the sites, respectively. The research of chironomid ecology, diversity and distribution continues, which will provide new data and information, but this first comprehensive checklist provides a good starting point for those studying Chironomidae or other freshwater Diptera, in Croatia and Europe.

Can phytoplankton blooming be harmful to benthic organisms? The toxic influence of Anabaena sp. and Chlorella sp. on Chironomus riparius larvae.

Cyanobacteria and microalgae are abundant biota groups in eutrophic freshwater ecosystems, serving as a food source for many aquatic organisms, including the larvae of non-biting midges (Chironomidae). Many species of cyanobacteria are toxin producers, which can act as stressors to other organisms. The present study aimed to analyze and compare the effects of dietary exposure to the common toxic cyanobacteria Anabaena sp. and non-toxic microalgae Chlorella sp. in Chironomus riparius larvae. Microcystin was detected and quantified in the methanolic extract of Anabaena sp. using the HPLC-DAD technique, and it was identified as microcystin-LR. Both Anabaena sp. and Chlorella sp. were suitable food sources to enable the survival of C. riparius larvae in laboratory conditions, causing negligible mortality and significant differences in the larval mass (ANOVA and Post hoc LSD test; p < 0.05) and hemoglobin concentration (Student's t-test; p < 0.05). Oxidative stress parameters such as advanced oxidation protein products (AOPP), thiobarbituric acid reactive substances (TBARS), catalase (CAT) and superoxide dismutase (SOD) activity, and DNA damage, were also investigated. One-way ANOVA, followed by the Post hoc LSD test, showed a significant increase in AOPP and CAT for the group of larvae fed with Chlorella sp. The same test showed moderate DNA damage in both groups of larvae, with greater damage in the group fed with Anabaena sp. Thus, Chlorella sp. and microcystin-LR producing Anabaena sp. are food sources that did not result in any drastic acute effect on the population level of C. riparius larvae. However, sub-individual-level endpoints revealed significant effects of the treatments, since they caused oxidative stress and DNA damage that may pose a danger to successive generations of test organisms.

Exposure to a microplastic mixture is altering the life traits and is causing deformities in the non-biting midge Chironomus riparius Meigen (1804).

The effect of microplastics (MP) exposure on the chironomid species Chironomus riparius Meigen, 1804 was investigated using the OECD sediment and water toxicity test. Chironomid larvae were exposed to an environmentally relevant low microplastics concentration (LC), a high microplastics concentration (HC) and a control (C). The LC was 0.007 g m-2 on the water surface + 2 g m-3 in the water column + 8 g m-2 in the sediment, and the HC was 10 X higher than this for each exposure. The size of the majority of the manufactured microplastic pellets varied between 20 and 100 µm. The MP mixture consisted of: polyethylene-terephtalate (PET), polystyrene (PS), polyvinyl-chloride (PVC) and polyamide (PA) in a ratio of 45%: 15%: 20%: 20%, respectively, for the sediment exposure; 100% polyethylene for the water column exposure; and 50% polyethylene: 50% polypropylene for the water surface exposure. Different endpoints were monitored, including morphological changes in the mandibles and mentums of 4th instar larvae, morphological changes in the wings, mortality, emergence ratio, and developmental time. A geometric morphometric analysis showed a tendency toward widening of the wings, elongation of the mentums and changing the shape of the mandibles in specimens exposed to both concentrations of microplastics. The development time of C. riparius was significantly prolonged by the MP treatment: 13.8 ± 0.5; 14.4 ± 0.6; and 15.3 ± 0.4 days (mean ± SD) in the C, LC, and HC, respectively. This study indicates that even environmentally relevant concentrations of MP mixture have a negative influence on C. riparius, especially at the larval stage.

Application of deep learning in aquatic bioassessment: Towards automated identification of non-biting midges.

Morphological species identification is often a difficult, expensive, and time-consuming process which hinders the ability for reliable biomonitoring of aquatic ecosystems. An alternative approach is to automate the whole process, accelerating the identification process. Here, we demonstrate an automatic machine-based identification approach for non-biting midges (Diptera: Chironomidae) using Convolutional Neural Networks (CNNs) as a means of increasing taxonomic resolution of biomonitoring data at a minimal cost. Chironomidae were used to build the automatic identifier, as a family of insects that are abundant and ecologically important, yet difficult and time-consuming to accurately identify. The approach was tested with 10 morphologically very similar species from the same genus or subfamilies, comprising 1846 specimens from the South Morava river basin, Serbia. Three CNN models were built utilizing either species, genus, or subfamily data. After training the artificial neural network, images that the network had not seen during the training phase achieved an accuracy of 99.5% for species-level identification, while at the genus and subfamily level all images were correctly assigned (100% accuracy). Gradient-weighted Class Activation Mapping (Grad-CAM) visualized the mentum, ventromental plates, mandibles, submentum, and postoccipital margin to be morphologically important features for CNN classification. Thus, the CNN approach was a highly accurate solution for chironomid identification of aquatic macroinvertebrates opening a new avenue for implementation of artificial intelligence and deep learning methodology in the biomonitoring world. This approach also provides a means to overcome the gap in bioassessment for developing countries where widespread use techniques for routine monitoring are currently limited.

A Multiparametric Approach to Cerium Oxide Nanoparticle Toxicity Assessment in Non-Biting Midges.

Cerium oxide nanoparticles (CeO2 NPs) are included in the Organisation for Economic Co-operation and Development (OECD) priority list of engineered nanomaterials for assessment of their environmental impact. The present study was carried out to assess the CeO2 NP toxicity to the freshwater midge Chironomus riparius larvae at concentrations of 2.5, 25, 250, and 2500 mg of CeO2 NP/kg of sediment. Experiments were designed to assess the prolonged exposure of midges to CeO2 NPs while adhering to OECD test guideline 218. The following parameters were investigated: CeO2 NP uptake by larvae, oxidative stress parameters, in vivo genotoxic effects, and life trait parameters. Inductively coupled plasma-mass spectrometry analysis showed a significant positive correlation between the concentration of CeO2 NPs in the sediment and its uptake by the larvae. No significant mortality was observed in C. riparius, and oxidative stress was not detected. The only significantly induced sublethal effect was genotoxicity, which began to manifest at a lowest-observed-effect concentration of 25 mg kg-1 of sediment and progressively increased at higher concentrations. Our results indicate that exposure to CeO2 NP-contaminated freshwater sediments does not pose a risk to chironomids at environmentally realistic concentrations. However, the significant accumulation of CeO2 NPs by chironomid larvae may pose a risk through trophic transfer to organisms further up the food chain. Environ Toxicol Chem 2019;39:131-140. © 2019 SETAC.

An environmentally relevant concentration of titanium dioxide (TiO2) nanoparticles induces morphological changes in the mouthparts of Chironomus tentans.

The present study was carried out in order to assess the influence of environmentally relevant concentrations of TiO2 nanoparticles (E171 human food grade) toxicity on the freshwater midge Chironomus tentans. Tested concentrations were 125, 250, 500, 1000, 2000 and 4000 mg of E171 TiO2 per 1 kg of sediment, for the experiment aiming at life trait toxicity observation; and 2.5, 25 and 250 mg of E171 TiO2 per 1 kg of sediment for the experiment aiming at mouthpart deformity observation. The experimental design was constructed for the sediment dwelling chironomid larvae according to OECD guidelines. For the first time, a geometric morphometric approach was used to assess the deformities in chironomid larvae as sublethal implications of nanoparticle exposure. The present study showed a concentration-response relationship between the TiO2 concentration in the substrate and the TiO2 intake. The mortality and emergence ratio was affected at concentrations >1000 mg/kg. Geometric Morphometrics revealed the tendency of the mentum teeth to narrow and elongate and the mandibles to widen, as well as the loss of the first inner tooth, with a TiO2 concentration rise. The variability of morphological changes observed in the mouthparts indicates that C. tentans could be used as a bioindicator in nano-TiO2 monitoring.

The response of chironomid taxonomy- and functional trait-based metrics to fish farm effluent pollution in lotic systems.

The lotic habitats affected by trout farm waste are colonized with a particular invertebrate community of which chironomids are the most abundant group. However, there is little information available regarding how chironomid community structures respond to this type of pollution at the highest taxonomic resolution. Eight fish farms, together with their lotic systems as recipients, were used to test the variability of the chironomid community and its surrogates (taxonomic and functional metrics) across spatially arranged sampling sites to form a gradual decrease in the trout farm influence. The self organizing map (SOM) classified six different types of chironomid communities which were characteristic for both the control and affected habitats. The species indicator analyses listed 32 taxa as positive indicators of water pollution. The SOM and Kruskal-Wallis test revealed that the pattern of chironomid community structure obtained was mainly driven by six environmental parameters (Altitude, conductivity, distance from the outlet, hardness, HN4-N, NO3-N). Categorical principal components analysis (CATPCA) derived three models for each type of biotic metric, in which for diversity-, taxonomy- and functional feeding group-based metrics, the first two dimensions explained 55.2%, 58.3% and 55.4%, of the total variance respectively for 315 sampling sites. According to this analysis, the total number of taxa (S), abundance and the Shannon-Wiener index (H') (as a diversity metric), as well as the proportion of Tanypodinae (as taxonomic group) and grazers/scraper (GRA) and gatherer collector (GAT)(as FFG metrics), were related to the outlet distance gradient, thus showing great potential to be used in the multimetric approach in bioassessment.

The best data design for applying the taxonomic distinctness index in lotic systems: A case study of the Southern Morava River basin.

The taxonomic distinctness (Δ+) index has been recognized as a robust measure to assess human impacts on marine biodiversity. However, its applicability in freshwater ecosystems has still not been confirmed. We aimed to propose the most suitable data design for calculating the Δ+ index for application in assessing anthropogenically caused degradation in lotic environments. We calculated the values of Δ+ based on different taxa groups and taxa resolutions, in order to examine its utility as a potential metric in bioassessment programs. We found that the exclusion of non-insect taxa and selected insect orders significantly increased the index sensitivity. Thus, we believe that an appropriate data design for Δ+ calculation based on macroinvertebrate assemblages is the main prerequisite for the effective estimation of degradation in lotic environments. In addition, we argue that a decrease in taxonomic resolution up to genus level is completely acceptable, as it results in only minor information loss. Bearing this in mind would significantly facilitate its application in rapid bioassessment programs. Despite the observed correlation, the utility of Δ+ as a potential bioassessment metric is rather limited, since its fails to detect fine differences in environmental stress, and instead only roughly distinguishes between two basic classes of degradation level, unimpacted and impacted.

The potential of chironomid larvae-based metrics in the bioassessment of non-wadeable rivers.

The chironomid community in non-wadeable lotic systems was tested as a source of information in the construction of biological metrics which could be used into the bioassessment protocols of large rivers. In order to achieve this, we simultaneously patterned the chironomid community structure and environmental factors along the catchment of the Danube and Sava River. The Self organizing map (SOM) recognized and visualized three different structural types of chironomid community for different environmental properties, described by means of 7 significant abiotic factors (a multi-stressor gradient). Indicator species analysis revealed that the chironomid taxa most responsible for structural changes significantly varied in their abundance and frequency along the established environmental gradients. Out of 40 biological metrics based on the chironomid community, the multilayer perceptron (MLP), an supervised type of artificial neural network, derived 5 models in which the abundance of Paratrichocladius rufiventis, Orthocladiinae, Cricotopus spp., Cricotopus triannulatus agg. and Cricotopus/Orthocladius ratio achieved a significant relationship (the r Pearson's linear correlation coefficient>0.7) with the multi stressor environment. The sensitivity analysis "partial derivatives" (PaD) method showed that all 5 biological metrics within the multi-stressor gradient were mostly influenced by dissolved organic carbon (DOC). Despite short and monotonous environmental gradients and the absence of reference conditions, the chironomid community structure and biological metrics predictably changed along the multistress range, showing a great potential for the bioassessment of large rivers.

In situ effects of titanium dioxide nanoparticles on community structure of freshwater benthic macroinvertebrates.

For the first time in the current literature, the effect of titanium dioxide (TiO2) nanoparticles on the community structure of macroinvertebrates has been investigated in situ. Macroinvertebrates were exposed for 100 days to an environmentally relevant concentration of TiO2 nanoparticles, 25 mg kg(-1) in sediment. Czekanowski's index was 0.61, meaning 39% of the macroinvertebrate community structure was affected by the TiO2 treatment. Non-metric multidimensional scaling (NMDS) visualized the qualitative and quantitative variability of macroinvertebrates at the community level among all samples. A distance-based permutational multivariate analysis of variance (PERMANOVA) revealed the significant effect of TiO2 on the macroinvertebrate community structure. The indicator value analysis showed that the relative frequency and abundance of Planorbarius corneus and Radix labiata were significantly lower in the TiO2 treatment than in the control. Meanwhile, Ceratopogonidae, showed a significantly higher relative frequency and abundance in the TiO2 treatment than in the control.