Freshwater Invertebrate Species Research Articles

Participatory hackathon to determine ecological relevant endpoints for a neurotoxin to aquatic and benthic invertebrates.

The aim of this study is twofold: i) to determine innovative yet sensitive endpoints for sulfoxaflor and ii) to develop best practices for innovative teaching in ecotoxicology. To this end, a group of 52 MSc students participated in an environmental hackathon, during which they did creative toxicity testing on 5 freshwater invertebrate species: Daphnia magna, Chironomus riparius, Asellus aquaticus, Lymnaea stagnalis, and Anisus vortex. Involving the students in an active learning environment stimulated increased creativity and productivity. In total, 28 endpoints were investigated, including standard endpoints (e.g., mortality) as well as biomechanistic and energy-related endpoints. Despite high variances in the results, likely linked to the limited lab experience of the students and interpersonal differences, a promising set of endpoints was selected for further investigation. A more targeted follow-up experiment focused on the most promising organism and set of endpoints: biomechanistic endpoints of C. riparius larvae. Larvae were exposed to a range of sulfoxaflor concentrations (0.90-67.2 μg/L) for 21 days. Video tracking showed that undulation and swimming were significantly reduced at 11.1 μg sulfoxaflor/L after 9 days of exposure, and an EC50 = 10.6 μg/L for mean velocities of the larvae in the water phase was found. Biomechanistic endpoints proved much more sensitive than mortality, for which an LC50 value of 116 μg/L was found on Day 9. Our results show that performing a hackathon with students has excellent potential to find sensitive endpoints that can subsequently be verified using more targeted and professional follow-up experiments. Furthermore, utilising hackathon events in teaching can increase students' enthusiasm about ecotoxicology, driving better learning experiences.

Multiple stressor effects of insecticide exposure and increased fine sediment deposition on the gene expression profiles of two freshwater invertebrate species

BackgroundFreshwater ecosystem degradation and biodiversity decline are strongly associated with intensive agricultural practices. Simultaneously occurring agricultural stressors can interact in complex ways, preventing an accurate prediction of their combined effects on aquatic biota. Here, we address the limited mechanistic understanding of multiple stressor effects of two globally important stressors, an insecticide (chlorantraniliprole), and increased fine sediment load and assessed their impact on the transcriptomic profile of two stream macroinvertebrates: the amphipod Gammarus pulex and the caddisfly Lepidostoma basale.ResultsWe identified mainly antagonistic stressor interactions at the transcriptional level, presumably because the insecticide adsorbed to fine sediment particles. L. basale, which is phylogenetically more closely related to the insecticide’s target taxon Lepidoptera, exhibited strong transcriptional changes when the insecticide stressor was applied, whereas no clear response patterns were observed in the amphipod G. pulex. These differences in species vulnerability can presumably be attributed to molecular mechanisms determining the cellular affinity toward a stressor as well as differential exposure patterns resulting from varying ecological requirements between L. basale and G. pulex. Interestingly, the transcriptional response induced by insecticide exposure in L. basale was not associated with a disruption of the calcium homeostasis, which is the described mode of action for chlorantraniliprole. Instead, immune responses and alterations of the developmental program appear to play a more significant role.ConclusionsOur study shows how transcriptomic data can be used to identify multiple stressor effects and to explore the molecular mechanisms underlying stressor-induced physiological responses. As such, stressor effects assessed at the molecular level can inform about modes of action of chemicals and their interplay with non-chemical stressors. We demonstrated that stressor effects vary between different organismic groups and that insecticide effects are not necessarily covered by their described mode of action, which has important implications for environmental risk assessment of insecticides in non-target organisms.

Toxicity of 6PPD-quinone to four freshwater invertebrate species

The antioxidant N-(1,3-Dimethylbutyl)-N′-phenyl-p- phenylenediamine (6PPD) is used to protect the rubber in tires from oxidation, which extends the life of the tire. When oxidized, 6PPD is transformed into 6PPD-quinone (6PPDQ). 6PPDQ, along with other tire ingredients, can enter aquatic ecosystems through the transport of tire wear particles in runoff during a precipitation event. The mass mortality of coho salmon following precipitation events in urban areas lead to the discovery that 6PPDQ is the likely cause due to coho salmon's relatively high sensitivity to 6PPDQ. The assessment of 6PPDQ toxicity to other aquatic species has expanded, but it has focused on fish. This study investigated the toxicity of 6PPDQ to four freshwater invertebrate species, larval burrowing mayfly (Hexagenia spp.), juvenile cladoceran (Daphnia magna), file ramshorn snail embryo (Planorbella pilsbryi), and adult washboard mussel (Megalonaias nervosa). For all four species, the highest concentration of 6PPDQ tested did not result in significant mortality. This translated into the determination of the highest concentration that did not cause significant mortality (NOEC) for Hexagenia spp., D. magna, P. pilsbryi, and M. nervosa of 232.0, 42.0, 11.7, and 17.9 μg/L, respectively. The data from this study indicate that freshwater invertebrates are not as sensitive to 6PPDQ as some salmonid species (e.g., coho salmon Oncorhynchus kisutch). This study also analyzed 6PPDQ in road runoff from around the city of Guelph in Ontario, Canada. 6PPQ was detected in all samples but the concentration was two orders of magnitude lower than the NOECs for the four tested species of freshwater invertebrate.

Are Antarctic aquatic invertebrates hitchhiking on your footwear?

The increasing number of humans who travel to and within Antarctic regions each year not only increases the risk of introducing non-native species but also of translocating native species within and between Antarctic biogeoregions and poses the potential for artificial, human-mediated introduction of native freshwater invertebrate species to newly ice-free areas of Antarctica. This study was designed to test the potential for transfer of native Antarctic freshwater invertebrates and their cysts on footwear. An average of 1.86 Anostraca cysts, 29.47 Copepoda cysts, 4.29 Nematoda, 0.40 Rotifera and 0.966 Tardigrada individuals per gram of sediment were found in samples taken from footwear after contact with freshwater sediment. The invertebrate cysts isolated from the samples that underwent drying at 27 °C were able to hatch in tap water, and representatives of the other systematic groups (Nematoda, Rotifera, and Tardigrada) also remained viable.

Potential risk to aquatic biota from aerial application of firefighting water additives

The frequency and severity of forest fires is increasing due to climate change. Consequently, there will be an increased use of forest firefighting additives, which increase the ability of water to extinguish fires and prevent reignition. Increased use will potentially result in increased exposure to aquatic ecosystems within forests. This study examined the toxicity of nine firefighting water additives that are currently on the market to three species of freshwater invertebrates that occupy different niches within freshwater ecosystems. The toxicity of the water additives varied up to three orders of magnitude. Pelagic and epibenthic invertebrates are affected at lower rates of application than endobenthic invertebrates. A field relevant application rate of three of the nine water additives tested represent a hazard to freshwater ecosystems under varies exposure scenarios represented by the depth of a theoretical water body (15–200 cm). This study highlights the importance of application buffers around water bodies and the selection of water additives that pose the lowest hazard to freshwater ecosystem, assuming that the efficacy of the additives in extinguishing fires is similar.

Investigating the potential impacts of coal ash runoff on the freshwater Seminole ramshorn snail (Planorbella duryi) under laboratory conditions

Coal fly ash is an industrial waste product generated by coal fired powerplants which has been shown to contain elevated concentrations of several toxic trace metals. When stored in landfills or other repositories, these trace metals can enter nearby surface waters via a number of routes including leaching or runoff. Our study examined 1) the presence and concentration of eleven trace elements in a range of lab-created coal ash leachate solutions at neutral pH using ICP-OES, 2) the physiological effects of these leachate solutions on a freshwater gastropod (Planorbella duryi), and 3) the ability of these trace metals to bioaccumulate in the tissues of exposed individuals. As, Cd, Cu, Mg, Mn, and Pb were detected in solutions at increasing concentrations concurrent with ash concentration. Exposure to leachates caused significant delays in embryonic development, reduced juvenile shell growth, decreases in egg and clutch production, and the display of avoidance behaviors. Tissues of exposed snails contained elevated concentrations of As, Cd, Cu, and Cr, with bioconcentration factors 177,550 times higher in cadmium and 85,468 times higher in arsenic in the highest treatment compared to control organisms. Our results highlight the potential harmful effects of coal ash leachates on a novel freshwater invertebrate species using several unique methodologies, providing key information regarding their potential impacts on surrounding aquatic ecosystems.

Antimicrobial peptides from freshwater invertebrate species: potential for future applications.

Invertebrates are a significant source of antimicrobial peptides because they lack an adaptive immune system and must rely on their innate immunity to survive in a pathogen-infested environment. Various antimicrobial peptides that represent major components of invertebrate innate immunity have been described in a number of investigations over the last few decades. In freshwater invertebrates, antimicrobial peptides have been identified in arthropods, annelids, molluscs, crustaceans, and cnidarians. Freshwater invertebrate species contain antimicrobial peptides from the families astacidin, macin, defensin, and crustin, as well as other antimicrobial peptides that do not belong to these families. They show broad spectrum activities greatly directed against bacteria and to a less extent against fungi and viruses. This review focuses on antimicrobial peptides found in freshwater invertebrates, highlighting their features, structure-activity connections, antimicrobial processes, and possible applications in the food industry, animal husbandry, aquaculture, and medicine. The methods for their synthesis, purification, and characterization, as well as the obstacles and strategies for their development and application, are also discussed.

Midbody-Localized Aquaporin Mediates Intercellular Lumen Expansion During Early Cleavage of an Invasive Freshwater Bivalve.

Intercellular lumen formation is a crucial aspect of animal development and physiology that involves a complex interplay between the molecular and physical properties of the constituent cells. Embryos of the invasive freshwater mussel Dreissena rostriformis are ideal models for studying this process due to the large intercellular cavities that readily form during blastomere cleavage. Using this system, we show that recruitment of the transmembrane water channel protein aquaporin exclusively to the midbody of intercellular cytokinetic bridges is critical for lumenogenesis. The positioning of aquaporin-positive midbodies thereby influences the direction of cleavage cavity expansion. Notably, disrupting cytokinetic bridge microtubules impairs not only lumenogenesis but also cellular osmoregulation. Our findings reveal a simple mechanism that provides tight spatial and temporal control over the formation of luminal structures and likely plays an important role in water homeostasis during early cleavage stages of a freshwater invertebrate species.

Bicarbonate alone does not totally explain the toxicity from major ions of coal bed derived waters to freshwater invertebrates

Concentrations of major ions in coal mine discharge waters and unconventional hydrocarbon produced waters derived from coal bed methane (CBM) production, are potentially harmful to freshwater ecosystems. Bicarbonate is a major constituent of produced waters from CBM and coal mining. However, little is known about the relative toxicity of differing ionic proportions, especially bicarbonate, found in these CBM waters. As all freshwater invertebrates tested are more acutely sensitive to sodium bicarbonate (NaHCO3) than sodium chloride (NaCl) or synthetic sea water, we tested the hypotheses that toxicity of CBM waters are driven by bicarbonate concentration, and waters containing a higher proportion of bicarbonate are more toxic to freshwater invertebrates than those with less bicarbonate. We compared the acute (96 h) lethal toxicity to six freshwater invertebrate species of NaHCO3 and two synthetic CBM waters, with ionic proportions representative of water from CBM wells across New South Wales (NSW) and Queensland (Qld), in Australia. The ranking of LC50 values expressed as total salinity was consistent with the hypotheses. However, when toxicity was expressed as bicarbonate concentration, the hypothesis that the toxicity of coal bed waters would be explained by bicarbonate concentration was not well supported, and other ionic components were either ameliorating or exacerbating the NaHCO3 toxicity. Our findings showed NaHCO3 was more toxic than NaCl and that the NaHCO3 proportion of synthetic CBM waters drives toxicity, however other ions are altering the toxicity of bicarbonate.

DNA metabarcoding unravels unknown diversity and distribution patterns of tropical freshwater invertebrates

Abstract Tropical freshwater invertebrate species are becoming extinct without being described, and effective conservation is hampered by a lack of taxonomic and distribution data. DNA metabarcoding is a promising tool for rapid biodiversity assessments that has never been applied to tropical freshwater invertebrates across large spatial and taxonomic scales. Here we use DNA metabarcoding to comprehensively assess the benthic freshwater invertebrate fauna of the Perak River basin, Malaysia. Specific objectives were to: (1) assess performance of two DNA metabarcoding protocols; (2) identify gaps in reference databases; (3) generate new data on species diversity and distribution; and (4) draw conclusions regarding the potential value of DNA metabarcoding in tropical freshwater conservation. Organisms were collected by hand and net at 34 sites and divided into small (retained in 0.5‐mm but passing through 1‐mm mesh) and large (retained in 1‐mm mesh) fractions, and a 313‐bp cytochrome c oxidase subunit I fragment amplified and sequenced using general Metazoa primers. Bioinformatic analysis resulted in 468 operational taxonomic units (~species) from 12 phyla. Only 29% of species could be assigned binominal names through matches to public sequence libraries, indicating varying levels of library completeness across Orders. Extraction of small‐fraction DNA with a soil kit resulted in a significantly higher species count than with a general kit, but this was not even across taxa. Metabarcoding (amplification) success rate, estimated via comparison to morphological identifications of the large‐fraction specimens, was high in most taxa analysed but low, for example, in ampullariid and viviparid gastropods. Conversely, a large proportion of species‐site records for Decapoda and Bivalvia came from metabarcoding only. Species richness averaged 29 ± 16 species per site, dominated by Diptera, Annelida, and Odonata, and was particularly high in tributaries of the mountainous Titiwangsa Range. At least eight species are new records for Malaysia, including the non‐natives Ferrissia fragilis (Gastropoda) and Dugesia notogaea (Platyhelminthes). Our study showed that DNA metabarcoding is generally more effective in detecting tropical freshwater invertebrate species than traditional morphological approaches, and can efficiently improve knowledge of distribution patterns and ranges of native and non‐native species. However, current gaps in reference databases, particularly for bioindicator taxa, such as the Plecoptera, Ephemeroptera, and Coleoptera, need to be addressed urgently.

Strategies of Invertebrate Osmoregulation: An Evolutionary Blueprint for Transmuting into Fresh Water from the Sea.

Early marine invertebrates like the Branchiopoda began their sojourn into dilute media some 500 million years ago in the Middle Cambrian. Others like the Mollusca, Annelida, and many crustacean taxa have followed, accompanying major marine transgressions and regressions, shifting landmasses, orogenies, and glaciations. In adapting to these events and new habitats, such invertebrates acquired novel physiological abilities that attenuate the ion loss and water gain that constitute severe challenges to life in dilute media. Among these taxon-specific adaptations, selected from the subcellular to organismal levels of organization, and constituting a feasible evolutionary blueprint for invading freshwater, are reduced body permeability and surface (S) to volume (V) ratios, lowered osmotic concentrations, increased osmotic gradients, increased surface areas of interface epithelia, relocation of membrane proteins in ion-transporting cells, and augmented transport enzyme abundance, activity, and affinity. We examine these adaptations in taxa that have penetrated into freshwater, revealing diversified modifications, a consequence of distinct body plans, morpho-physiological resources, and occupation routes. Contingent on life history and reproductive strategy, numerous patterns of osmotic regulation have emerged, including intracellular isosmotic regulation in weak hyper-regulators and well-developed anisosmotic extracellular regulation in strong hyper-regulators, likely reflecting inertial adaptations to early life in an estuarine environment. In this review, we address osmoregulation in those freshwater invertebrate lineages that have successfully invaded this biotope. Our analyses show that across 66 freshwater invertebrate species from six phyla/classes that have transmuted into freshwater from the sea, hemolymph osmolalities decrease logarithmically with increasing S:V ratios. The arthropods have the highest osmolalities, from 300 to 650 mOsmoles/kg H2O in the Decapoda with 220-320 mOsmoles/kg H2O in the Insecta; osmolalities in the Annelida range from 150 to 200 mOsmoles/kg H2O, and the Mollusca showing the lowest osmolalities at 40-120 mOsmoles/kg H2O. Overall, osmolalities reach a cut-off at ∼200 mOsmoles/kg H2O, independently of increasing S:V ratio. The ability of species with small S:V ratios to maintain large osmotic gradients is mirrored in their putatively higher Na+/K+-ATPase activities that drive ion uptake processes. Selection pressures on these morpho-physiological characteristics have led to differential osmoregulatory abilities, rendering possible the conquest of freshwater while retaining some tolerance of the ancestral medium.

Effects of Diclofenac on the Reproductive Health, Respiratory Rate, Cardiac Activity, and Heat Tolerance of Aquatic Animals.

Diclofenac is an important pharmaceutical present in the water cycle of wastewater treatment and one of the most distributed drugs in aquatic ecosystems. Despite the great interest in the fate of diclofenac in freshwaters, the effects of environmentally relevant concentrations on invertebrates are still unclear. Two species of freshwater invertebrates, the amphipod Gmelinoides fasciatus and the bivalve mollusk Unio pictorum, were exposed to diclofenac concentrations of 0.001-2 μg/L (environmentally relevant levels) for 96 h. A set of biological endpoints (survival, fecundity, embryo abnormalities, respiration and heart rates, heat tolerance, and cardiac stress tolerance) were estimated in exposed invertebrates. Effects of diclofenac on amphipod metabolic rate and reproduction (number and state of embryos) and adaptive capacity (cardiac stress tolerance) in both species were evident. The oxygen consumption of amphipods exposed to diclofenac of 0.1-2 μg/L was 1.5-2 times higher than in the control, indicating increased energy requirements for standard metabolism in the presence of diclofenac (>0.1 μg/L). The heart rate recovery time in mollusks after heating to critical temperature (30 °C) was 1.7 and 9 times greater in mollusks exposed to 0.1 and 0.9 μg/L, respectively, than in the control (24 min). A level of diclofenac >0.9 μg/L adversely affected amphipod embryos, leading to an increase in the number of embryos with impaired development, which subsequently died. Thus, the lowest effective concentration of diclofenac (0.1 μg/L) led to increased energy demands of animals while reducing cardiac stress tolerance, and at a level close to 1 μg/L reproductive disorders (elevated mortality of the embryos) occurred. Environ Toxicol Chem 2022;41:677-686. © 2021 SETAC.

Some alien species of aquatic invertebrates in the water bodies of the nature reserve fund of the Kyiv metropolis

The invasion of alien species of freshwater invertebrates and the associated changes in the faunistic composi- tion of local ecosystems are a serious threat to the conservation of biodiversity. In this work, we supplement the existing list of alien species with information about three more species of non-native freshwater invertebrates, which have formed stable populations in water bodies of Ukraine. These are two representatives of flatworms: Girardia tigrina (Girard, 1850) and Stenostomum saliens Kepner & Carter, 1931, as well as a representative of copepods, Eucyclops roseus Ishida, 1997.

Population genetic structure and connectivity in three montane freshwater invertebrate species (Ephemeroptera, Plecoptera, Amphipoda) with differing life cycles and dispersal capabilities

Abstract Dispersal abilities and the resulting levels of connectivity between streams influence population structure in aquatic organisms. We investigated how different dispersal and life‐history traits affect gene flow and population structure in three aquatic invertebrate species in a central European mountain range. We used microsatellite markers and mitochondrial cytochrome oxidase I barcode data to assess small‐scale population structure and connectivity of the mayfly Baetis alpinus, the stonefly Brachyptera seticornis, and the amphipod Gammarus fossarum in seven streams within Bavarian Forest National Park, Germany. Significant population structure within the study area was detected in B. alpinus and G. fossarum, whereas B. seticornis had signatures of panmixia. In G. fossarum, that structure corresponded to stream topography, while in B. alpinus it did not. The Bavarian Forest range appears to be a contact zone for different mitochondrial lineages of B. alpinus and G. fossarum. Limited geneflow between sample sites in B. alpinus can be explained by the species’ short adult lifespans, which restricts dispersal between sites. Since imagines are able to disperse via adult flight population structure is, however, decoupled from stream topology. Longer‐lived B. seticornis is better able to maintain geneflow between sample sites. In flightless G. fossarum, population structure corresponding to stream topology can be explained by limited dispersal capabilities between streams. Dispersal ability alone is insufficient to predict or explain patterns of gene flow and connectivity for individual species, and should be examined together with life‐history traits when assessing population and community structure.

Synthesis of New Betaine-Based Ionic Liquids by Using a “One-Pot” Amidation Process and Evaluation of Their Ecotoxicity through a New Method Involving a Hemocyte-Based Bioassay

A new class of betaine-based ionic liquids (ILs) have been synthesized. Betaine is transformed into betaine amides with various anions. These amides were prepared at first through a classical four-step method; the synthesis was then improved by reducing the number of steps using a one-pot strategy for amidation. The final yields were good to high, and the various structures were fully characterized to valorize them in many domains such as catalysis, extraction, or dissolution of biopolymers. Next, to determine their ecotoxicity, a hemocyte-based bioassay was used. This new method to evaluate the ecotoxicity of ILs was also applied to other ILs [tetrabutyl-ammonium (TBA), tetrabutyl-phosphonium (TBP), and betaine- and cholinium-based ILs] to prove its efficiency and its complementarity toward other ecotoxicological assessment methods. TBA- and TBP-based ILs induced limited effects on zebra mussel hemocytes, with TBP-ILs proving to be generally less toxic than TBA-based ones. On the contrary, betaine- and cholinium-based IL exposure led to more acute toxicity with a noticeable effect associated with the cation carbon chain length. Overall results also showed differential toxicity of ILs according to the nature of the anion in the assemblage, with lactate being the less biologically reactive anion. The results of this study thus reveal the importance of the nature of both cations and anions which constitute the IL structures and can strongly, sometimes synergistically, influence the IL toxicity. The possible interactions between the cationic and anionic parts reveal the complexity of IL toxicity prediction, thereby arousing interest in developing innovative strategies for the risk assessment of these molecular assemblages. In this context, the proposed hemocyte-based bioassay proved its efficiency and sensitivity in scoring and ranking IL toxicity according to their effects in the immunocompetent cells of an environmentally sentinel bivalve. This alternative bioassay could represent a complementary tool to more traditional and standard bioassays to complete the toxicity screening of substances such as ILs or new chemicals synthesized under green chemistry principles and notably the limitation of environmental toxicity by investigating the acute and functional effects on a wild and relevant freshwater invertebrate species.

Do bio-insecticides affect only insect species? Behavior, regeneration, and sexual reproduction of a non-target freshwater planarian.

Bio-insecticides have been increasingly used worldwide as ecofriendly alternatives to pesticides, but data on their effects in non-target freshwater organisms is still scarce and limited to insects. The aim of this study was to determine the lethal and sub-lethal effects of the bio-insecticides Bac Control (based on Bacillus thuringiensis kurstaki-Btk) and Boveril (based on Beauveria bassiana-Bb) on regeneration, behavioral, and reproductive endpoints of the freshwater planarian Girardia tigrina. The estimated LC50-48h were > 800 mg a.i./L for Btk and 60.74 mg a.i./L for Bb. In addition, exposure to Btk significantly decreased locomotion and feeding activities of planarians (lowest observed effect concentration (LOEC) of 12.5 mg a.i./L Btk) and fecundity rate (LOEC = 3.12 mg a.i./L Btk), whereas exposure to Bb significantly delayed regeneration (LOEC = 0.75 mg a.i./L Bb) and decreased fecundity rate (1.5 mg a.i./L Bb) of planarians. Thus, both bio-insecticides induced deleterious sub-lethal effects on a non-insect freshwater invertebrate species. However, only Bb-based formulation affected the survival, fecundity rate, and regeneration at concentrations below the maximum predicted environmental concentration (PEC = 247 mg/L). Thus, care should be taken when using such formulations as alternatives to chemical insecticides near aquatic ecosystems.

Effects of diclofenac on the swimming behavior and antioxidant enzyme activities of the freshwater interstitial crustacean Bryocamptus pygmaeus (Crustacea, Harpacticoida)

Diclofenac (DCF) is one of the most widespread pharmaceutical compounds found in freshwaters as a pseudo-persistent pollutant due to its continuous release from point and diffuse sources, being its removal in Wastewater Treatment Plants incomplete. Moreover, DCF is particularly persistent in interstitial habitats and potentially toxic for the species that spend their whole life cycle among the same sediment grains. This study is aimed at offering a first contribution to the assessment of DCF effects on freshwater invertebrate species living in the interstitial habitats of springs, rivers, lakes and groundwaters. The Crustacea Copepoda are one of the main components of the freshwater interstitial communities, with the primacy taken by the worm-like and small-sized harpacticoids. A sub-lethal concentration of 50 μg L-1 DCF significantly affected six out of the eight behavior parameters of the burrower/interstitial crustacean harpacticoid Bryocamptus pygmaeus recorded by video tracking analysis. DCF exposure reduced swimming speed, swimming activity, exploration ability and thigmotaxis, and increased swimming path tortuosity. The biochemical approach revealed a reduced level of the mitochondrial superoxide dismutase 2 in individuals exposed to DCF. It could be explained by a decline in mitochondrial performance or by a reduced number of functional mitochondria. Since mitochondrial dysfunction may determine ATP reduction, it comes that less energy is produced for maintaining the cell functions of the DCF-exposed individuals. In addition, the increasing energy demand for the detoxification process further contributes to decrease the total energetic budget allocated for other physiological activities. These observations can explain the changes we have observed in the swimming behavior of the copepod B. pygmaeus.

Shedding light on toxicity of SARS-CoV-2 peptides in aquatic biota: A study involving neotropical mosquito larvae (Diptera: Culicidae)

Knowledge about how the COVID-19 pandemic can affect aquatic wildlife is still extremely limited, and no effect of SARS-CoV-2 or its structural constituents on invertebrate models has been reported so far. Thus, we investigated the presence of the 2019-new coronavirus in different urban wastewater samples and, later, evaluated the behavioral and biochemical effects of the exposure of Culex quinquefasciatus larvae to two SARS-CoV-2 spike protein peptides (PSPD-2002 and PSPD-2003) synthesized in our laboratory. Initially, our results show the contamination of wastewater by the new coronavirus, via RT-qPCR on the viral N1 gene. On the other hand, our study shows that short-term exposure (48 h) to a low concentration (40 μg/L) of the synthesized peptides induced changes in the locomotor and the olfactory-driven behavior of the C. quinquefascitus larvae, which were associated with increased production of ROS and AChE activity (cholinesterase effect). To our knowledge, this is the first study that reports the indirect effects of the COVID-19 pandemic on the larval phase of a freshwater invertebrate species. The results raise concerns at the ecological level where the observed biological effects may lead to drastic consequences.

The toxic influence and biodegradation of carbon nanofibers in freshwater invertebrates of the families Gammaridae, Ephemerellidae, and Chironomidae

Carbon nanofibers (CNFs) are widely used in consumer products today. In this study, we assessed the effects of CNFs on the digestive system of three freshwater invertebrate species (Gammaridae, Ephemerellidae, and Chironomidae). The aquatic insects Diamesa sp., Drunella cryptomeria, and Gammarus suifunensis were incubated with the CNFs at the concentration of 100 mg/L during the 7-days period. Histological examination of the whole specimens and the longitudinal sections revealed no toxic effects of CNFs. However, a noticeable change in the structure of the CNFs accumulated in the intestines of the aquatic insects was found by Raman spectroscopy. The registered decrease in the relative proportion of amorphous carbon included in the CNF sample was found in the intestines of Diamesa sp. and D. cryptomeria. The registered effect can indicate a biodegradation of amorphous carbon in the digestive tract of these two insect species. In contrast, the decrease of highly structured carbons and the decrease of G-bonds intensity were registered in the digestive tract of G. suifunensis. This observation demonstrates the partial biodegradation of CNFs in the digestive tract of G. suifunensis.

AQUALIFE Software: A New Tool for a Standardized Ecological Assessment of Groundwater Dependent Ecosystems

We introduce a suite of software tools aimed at investigating multiple bio-ecological facets of aquatic Groundwater Dependent Ecosystems (GDEs). The suite focuses on: (1) threats posed by pollutants to GDE invertebrates (Ecological Risk, ER); (2) threats posed by hydrological and hydromorphological alterations on the subsurface zone of lotic systems and groundwater-fed springs (Hydrological-Hydromorphological Risk, HHR); and (3) the conservation priority of GDE communities (Groundwater Biodiversity Concern index, GBC). The ER is assessed by comparing tolerance limits of invertebrate species to specific pollutants with the maximum observed concentration of the same pollutants at the target site(s). Comparison is based on an original, comprehensive dataset including the most updated information on tolerance to 116 pollutants for 474 freshwater invertebrate species. The HHR is assessed by accounting for the main direct and indirect effects on both the hyporheic zone of lotic systems and groundwater-fed springs, and by scoring each impact according to the potential effect on subsurface invertebrates. Finally, the GBC index is computed on the basis of the taxonomical composition of a target community, and allows the evaluation of its conservation priority in comparison to others.