Water Flea Daphnia Magna Research Articles

Short-term and long-term effects of microplastics and organic UV-filters on the invertebrate model species Daphnia magna.

There is an ongoing debate regarding the role of microplastics (MPs) in enhancing the effects of various chemical compounds, highlighting the need for more detailed analyses. In this study, neonates of the water flea (Daphnia magna) were exposed to polystyrene MPs (PS-MPs; 3µm; exposure concentration, 1.25mg/L), a mixture of seven organic UV-filters (avobenzone, ethylhexyl triazone, homosalate, iscotrizinol, octinoxate, octisalate, and octocrylene; each at a low environmental concentration of 200ng/L), or the combination of both pollutants for 3, 7, or 21days. Results showed that PS-MPs alone decreased the body size of daphnids, while all treatments increased heart rate by the end of the 21-day exposure. On days 3 and 21, both PS-MPs and PS-MPs + UV-filters reduced swimming speed and total distance travelled. Additionally, PS-MPs increased the time of the first egg production, but decreased the egg number in the first production, total egg number, maximum egg number, and total neonate number during the 21-day treatment. Similarly, UV-filters or the combined pollutants increased the time of the first egg production and decreased the total neonate number. All treatments increased multixenobiotic resistance activity on days 3 and 7, while only UV-filters elevated CYP450 activity on day 3. PS-MPs or combined pollutants increased GST activity during early exposure but showed no effect on day 21. CAT activity was also affected by treatments in a time-dependent manner. These findings demonstrate that chronic exposure to PS-MPs and UV-filters, applied individually or in combination at a low environmental concentration, moderately impacts development, heart rate, and swimming activity in D. magna, while significantly altering reproduction and key cellular functions such as membrane transport activity, metabolism, and antioxidant defense. Co-exposure did not reveal a clear pattern of synergism or antagonism, suggesting that joint toxicity risks of these xenobiotics typically emerge at concentrations higher than low environmental levels. Future studies should explore potential interactions more thoroughly and assess transgenerational effects on reproduction and cellular defense pathways.

Transgenerational Effects of Atrazine on Daphnia magna Based on Life-History Traits and Population Dynamics.

Atrazine is a predominant herbicide globally, and its residues are commonly found in natural water bodies due to its extensive use. Atrazine is known for its detrimental effects on the reproductive abilities of aquatic plants and animals. Our study explored the impact of maternal exposure to atrazine on the survival and performance of offspring using the water flea Daphnia magna as a model organism. We investigated how the life-history traits and population dynamics of maternal D. magna and their offspring are influenced by atrazine. We hypothesized that exposure to atrazine in mothers would negatively affect the survival and performance of their offspring. The findings revealed that short-term exposure to atrazine adversely affects fitness-related traits, manifesting as decreased survivorship and reduced fecundity. Offspring from atrazine-exposed mothers displayed a limited recovery in fitness; no mortality was observed during the 14-day experiment, yet fecundity remained significantly reduced. At the population level, maternal exposure to atrazine led to a decreased population carrying capacity. The notable maternal effects on offspring reproduction underscore the long-term toxicity of atrazine to D. magna populations, suggesting prolonged consequences even after the cessation of atrazine pollution. This study underscores the transgenerational effects of atrazine, highlighting its persistent threat to D. magna populations.

Increased adsorption of diflubenzuron onto polylactic acid microplastics after ultraviolet weathering can increase acute toxicity in the water flea (Daphnia magna)

The ultraviolet (UV) weathering of microplastics (MPs) can lead to higher adsorption of harmful contaminants, thus increasing the potential risks of their combined effects. Because biodegradable MPs are more susceptible to UV weathering than conventional MPs, concerns have arisen about their ecological toxicity and environmental impact. Therefore, this study investigated the mechanisms associated with the adsorption of the pesticide diflubenzuron (DFB) onto polylactic acid (PLA) MP particles after UV weathering and the acute effects (48h) of their combination on the water flea Daphnia magna. These effects were also compared with those of the conventional MP polyethylene terephthalate (PET). UV weathering led to a greater number of cracks and pores in the PLA particles compared to PET, as well as a higher number of oxygen-based functional groups and a larger surface area. These surface changes in UV-weathered PLA particles promoted higher DFB adsorption, which in turn led to stronger acute toxicity for D. magna compared to UV-weathered PET particles. Combined exposure to 25ngL-1 DFB and both UV-weathered and non-UV-weathered MPs significantly reduced the chitin content in D. magna, while combined exposure to 12.5ngL-1 DFB and the MPs increased the chitin content. This effect was more pronounced for UV-weathered PLA exposure than UV-weathered PET exposure. The expression of the genes for chitinase and endocrine glycoprotein, both of which are closely associated with the toxic mechanisms of DFB, showed no significant changes with the combination of 25ngL-1 DFB and non-UV-weathered MPs but were significantly downregulated after UV weathering. Overall, the UV weathering of PLA promoted the adsorption of DFB, thus increasing its toxic effects. Our findings demonstrate the importance of considering the effects of UV weathering and interactions with environmental pollutants when assessing the ecological risks associated with biodegradable MPs.

Uptake, removal and trophic transfer of fluorescent polyethylene microplastics by freshwater model organisms: the impact of particle size and food availability

As an emerging contaminant, microplastics (MPs) are widely distributed in freshwater ecosystems and pose potential threats to aquatic organisms, attracting significant attention from both the scientific community and the general public. However, there is still uncertainty regarding the mechanisms of MPs transfer within aquatic biota and how particle size and food availability influence their transport patterns. In this study, zebrafish (Danio rerio) were selected as a model organism to investigate the uptake and elimination of fluorescent polyethylene (PE) MPs under different exposure scenarios (waterborne or trophic transfer, with or without food) and varying particle sizes (ranging from 10-300 μm at concentrations of 0.1, 2, and 300 mg/L). Additionally, water fleas (Daphnia magna) were provided as prey for the fish. The dynamic accumulation of PE-MPs sized between 10-20 μm at a concentration of 25 mg/L by daphnia was also determined along with its impact on animal feeding behavior. The results demonstrated that both organisms were capable of ingesting PE-MPs during exposures lasting up to 24 hours for daphnia and up to 72 hours for zebrafish. Furthermore, rapid elimination rates were observed within just 30 minutes for daphnia and between 6-12 hours for zebrafish. The presence of food reduced MPs uptake and removal by daphnia but significantly increased MP elimination by fish. Zebrafish showed a preference for ingesting larger-sized MPs that they could easily recognize; however, trophic transfer from daphnia to fish was found to be the primary route of ingestion specifically for PE-MPs sized between 10-20 μm. The findings suggest that while fish directly ingest fewer invisible MPs from the water column, they still accumulate these particles through predation on contaminated prey organisms. Therefore, it is imperative to prioritize the ecological risks associated with the transfer of MPs from zooplankton to fish.

Disposable face masks into aquatic media: Chemical and biological testing of the released compounds during the leaching process

The present study investigated the fate, and the biological effects posed by the presence of Disposable Face Masks (DFMs) into fresh- and saltwater media, using both chemical and biological testing. To this end, slightly fragmented DFMs were maintained in tanks with artificial sea water (ASW) or dH2O (DFMASW and DFMdH2O, respectively) for a period of 20 days (under continuous agitation, oxygen supply, and light/dark ration 1:1) to simulate both fresh- and saltwater natural conditions. Thereafter, DFMs leaching substances were determined, before proceeding to biological testing with the use of the marine bacterium Aliivibrio fischeri (Bioluminescence Inhibition assay), the fresh- and saltwater algal species Chlorococcum sp. and Tetraselmis suecica (algal bioassays), as well as the fairy shrimp Thamnocephalus platyurus, the water flea Daphnia magna, and the rotifer Brachionus calyciflorus (acute toxicity screening tests, in terms of microbiotest). According to the results, once into aquatic media (DFMASW and DFMdH2O) DFMs are subjected to degradation, leading to the release of organic, inorganic, and polymeric compounds (PP microfibers). Considering that possible interactions of the leaching substances could differentially affect the aquatic biota, the present study showed that DFMs leaching substances could be harmful to the fairy shrimp T. platyurus, and the water flea D. magna, with slight to non-toxic effects to be observed in case of Chlorococcum sp. and Tetraselmis suecica, the marine bacterium Aliivibrio fischeri, and the rotifer B. calyciflorus. The present findings showed that the DFMs improper disposal into fresh- and/or saltwater media, followed by their degradation and leaching processes, could lead to the release of substances of great environmental concern, thus promoting awareness about their proper handling and management, as well as the long-term monitoring of their environmental risk.

Effects of New Btk-Based Formulations BLB1 and Lip on Aquatic Non-Target Organisms.

Integrated pest management based on the use of biopesticides is largely applied. Experimental bioassays are critical to assess biopesticide biosafety at the ecotoxicological level. In this study, we investigated the effects of the new Bacillus thuringiensis subsp. kurstaki (Btk)-formulated-based biopesticides BLB1 and Lip, efficiently tested in field assays (IPM-4-CITRUS EC project no. 734921) on two aquatic non-target organisms, precisely the water flea Daphnia magna and the bioluminescent bacteria Aliivibrio fischeri. Acute toxicity studies, carried out in a comparative manner with Delfin® as the reference bioproduct and the lactose-based Blank formulation, show that no significant toxicity was observed up to 1 g/L. Our results indicated that BLB1- and Lip-formulated new bioproducts are far less toxic than the Delfin® reference bioproduct.

MicroRNA miR-210 Modulates the Water Flea Daphnia magna Response to Cyanobacterial Toxicity.

As a key form of post-transcriptional regulation, microRNAs (miRNAs) regulate gene expression by binding to target mRNAs, leading to mRNA decay or translational repression. Recently, the role of miRNAs in the response of aquatic organisms to environmental stressors has emerged. Daphnia, widely distributed cladocerans, play a crucial role in aquatic ecosystems. Cyanobacterial blooms often cause Daphnia populations to decrease, thereby disrupting ecosystem functionality and water quality. However, the post-transcriptional mechanisms behind Daphnia's response to toxic cyanobacteria are insufficiently understood. This study investigated the role of miR-210, a multifunctional miRNA involved in stress response and toxicity pathways, and its target genes (MLH3, CDHR5, and HYOU1) in two Daphnia magna clones exposed to toxic Microcystis aeruginosa. Results showed that M. aeruginosa inhibited somatic growth rates, led to microcystin accumulation, caused abnormal ultrastructural alterations in the digestive tract, and induced DNA damage in both clones. Notably, exposure significantly increased miR-210 expression and decreased the expression of its target genes compared with the controls. We identified miR-210s regulation on clonal-tolerance variations in D. magna to M. aeruginosa, emphasizing miRNAs' contribution to adaptive responses. Our work uncovered a novel post-transcriptional mechanism of cyanobacterial impact on zooplankton and provided essential insights for assessing cyanobacterial toxicity risks.

Comparing the effects of pristine and UV–VIS aged microplastics: Behavioural response of model terrestrial and freshwater crustaceans

Physico-chemical properties of microplastics (MPs) change during weathering in the environment. There is a lack of knowledge about the effects of such environmentally relevant MPs on organisms. We investigated: 1) the physico-chemical changes of MPs due to UV-VIS weathering, and 2) compared the effect of pristine and aged MPs on the behaviour of the water flea Daphnia magna and terrestrial crustacean Porcellio scaber. Dry powders of MPs were produced from widely used polymer types: disposable three-layer polypropylene (PP) medical masks (inner, middle and outer), polyester textile fibres, car tires and low-density polyethylene (LDPE) bags and were subjected to accelerated ultraviolet-visible (UV-VIS) ageing. Our results show that the extent of transformation depends on the type of polymer, with PP showing the most changes, followed by LDPE, textile fibres and tire particles. Obvious fragmentation was observed in PP and textile fibres. In the case of PP, but not polyester textile fibres, changes in FTIR spectra and surface properties were observed. Tire particles and LDPE did not change in size, but clear changes were observed in their FTIR spectra. Most MPs, aged and pristine, did not affect the swimming of daphnids. The only effect observed was a significant increase in path length and swimming speed for the pristine tire particles when the recording was done with particles remaining in the wells. After transfer to a clean medium, this effect was no longer present, suggesting a physical rather than chemical effect. Similarly, woodlice showed no significant avoidance response to the MPs tested, although there was a noticeable trend to avoid soils contaminated with pristine polyester textile fibers and preference towards the soils contaminated with aged MP of the middle mask layer. Overall, the apparent changes in physico-chemical properties of MPs after accelerated ageing were not reflected in their effects on woodlice and daphnids.

Comparative transcriptomics suggests a potential realizator gene for carapace expansion in longtail tadpole shrimp, Triops longicaudatus (Branchiopoda: Notostraca).

The origin of morphological innovation has been extensively studied within evolutionary developmental biology (evo-devo). Recent studies have demonstrated that the developmental module for double-layered epithelial outgrowths is conserved between the insect wings and branchiopod crustacean carapace, thereby introducing homology among these diverse structures. However, evo-devo studies on the branchiopod crustacean carapace have been primarily limited to a single species, the water flea Daphnia magna, leaving the gene regulatory network governing carapace development not comprehensively understood. Furthermore, realizator genes downstream of the character identity mechanism (ChIM) for bilayered epithelial development remain inadequately described. In this study, we analyzed tissue-specific transcriptional profiles in the developing longtail tadpole shrimp, Triops longicaudatus. We observed significant upregulation of papilin in the carapace-bearing head, along with its expression in both the carapace and the trunk limb lobes. Based on these results, we hypothesize that differential expression of papilin is involved in the disproportional growth of Triops carapace. Our findings will contribute to elucidating the diversification of double-layered epithelial outgrowths across distant arthropod lineages.

Exploring the Sublethal Impacts of Cu and Zn on Daphnia magna: a transcriptomic perspective

Metal contamination of aquatic environments remains a major concern due to their persistence. The water flea Daphnia magna is an important model species for metal toxicity studies and water quality assessment. However, most research has focused on physiological endpoints such as mortality, growth, and reproduction in laboratory settings, as well as neglected toxicogenomic responses. Copper (Cu) and zinc (Zn) are essential trace elements that play crucial roles in many biological processes, including iron metabolism, connective tissue formation, neurotransmitter synthesis, DNA synthesis, and immune function. Excess amounts of these metals result in deviations from homeostasis and may induce toxic responses. In this study, we analyzed Daphnia magna transcriptomic responses to IC5 levels of Cu (120 µg/L) and Zn (300 µg/L) in environmental water obtained from a pristine lake with adjusted water hardness (150 mg/L CaCO3). The study was carried out to gain insights into the Cu and Zn regulated stress response mechanisms in Daphnia magna at transcriptome level. A total of 2,688 and 3,080 genes were found to be differentially expressed (DEG) between the control and Cu and the control and Zn, respectively. There were 1,793 differentially expressed genes in common for both Cu and Zn, whereas the number of unique DEGs for Cu and Zn were 895 and 1,287, respectively. Gene ontology and KEGG pathways enrichment were carried out to identify the molecular functions and biological processes affected by metal exposures. In addition to well-known biomarkers, novel targets for metal toxicity screening at the genomic level were identified.

Ecotoxicological Assessment of Ibayo River in Balanga City, Bataan using Water Fleas (Daphnia Magna): A Basis for Intervention

The water quality of surface waters is degrading tremendously due to human interventions and development. Especially in the Philippines, water pollution has become a prominent problem in urbanized areas. Thus, this study aims to investigate the ecotoxicological effects of the Ibayo River in Balanga City, Bataan using water fleas (daphnia magna), a type of zooplankton living in freshwater areas. Daphnids are used as indicators in toxicity testing, and in assessing water quality. The survival rate of water fleas is utilized to examine the properties of the examined. As the water fleas were subjected to the river water samples and their dilutions for a week, results showed a high survival rate in all samples. It also exhibited no significant difference in the sample places Meanwhile, laboratory analysis revealed that there is an organic pollution as an excessive amount of biological oxygen demand (BOD) and lesser dissolved oxygen from the sample places were found. Analysis for chlorine and pH level indicates a n ormal quantity. Overall, there are chemicals in the Ibayo river that may kill aquatic organisms residing in it, but the long-term survival of these species is still questionable.

Coinfection frequency in water flea populations is a mere reflection of parasite diversity

In nature, parasite species often coinfect the same host. Yet, it is not clear what drives the natural dynamics of coinfection prevalence. The prevalence of coinfections might be affected by interactions among coinfecting species, or simply derive from parasite diversity. Identifying the relative impact of these parameters is crucial for understanding patterns of coinfections. We studied the occurrence and likelihood of coinfections in natural populations of water fleas (Daphnia magna). Coinfection prevalence was within the bounds expected by chance and parasite diversity had a strong positive effect on the likelihood of coinfections. Additionally, coinfection prevalence increased over the season and became as common as a single infection. Our results demonstrate how patterns of coinfection, and particularly their temporal variation, are affected by overlapping epidemics of different parasites. We suggest that monitoring parasite diversity can help predict where and when coinfection prevalence will be high, potentially leading to increased health risks to their hosts.

Pollution offsets the rapid evolution of increased heat tolerance in a natural population

Despite the increasing evidence for rapid thermal evolution in natural populations, evolutionary rescue under global warming may be constrained by the presence of other stressors. Highly relevant in our polluted planet, is the largely ignored evolutionary trade-off between heat tolerance and tolerance to pollutants. By using two subpopulations (separated 40years in time) from a resurrected natural population of the water flea Daphnia magna that experienced a threefold increase in heat wave frequency during this period, we tested whether rapid evolution of heat tolerance resulted in reduced tolerance to the widespread metal zinc and whether this would affect heat tolerance upon exposure to the pollutant. Our results revealed rapid evolution of increased heat tolerance in the recent subpopulation. Notably, the sensitivity to the metal tended to be stronger (reduction in net energy budget) or was only present (reductions in heat tolerance and in sugar content) in the recent subpopulation. As a result, the rapidly evolved higher heat tolerance of the recent subpopulation was fully offset when exposed to zinc. Our results highlight that the many reports of evolutionary rescue to global change stressors may give a too optimistic view as our warming planet is polluted by metals and other pollutants.

The analysis of inducible family members in the water flea Daphnia magna led to the identification of an uncharacterized lineage of heat shock protein 70

To explore the function and evolutionary relationships of inducible heat shock protein 70 (Hsp70) in Daphnia magna, cDNAs of four Hsp70 family members (DmaHsp70, DmaHsp70-2, DmaHsp70-12, DmaHsp70-14) were cloned. While all DmaHsp70s possess three function domains, it is noteworthy that only DmaHsp70 ends with a "EEVD" motif. Phylogenetic analysis indicates that the Hsp70-12 lineage is distanced from the rest, and therefore it is an uncharacterized lineage of Hsp70. The differences in isoelectric point and 3-dimensional (3D) conformation of the N-terminal nucleotide binding domain (NBD) of DmaHsp70s further support the theory. DmaHsp70s exhibit varied motif distribution patterns and the logo sequences of motifs have diverse signature characteristics, indicating that different mechanisms are involved in the regulation of ATP binding and hydrolysis for the DmaHsp70s. Protein-protein network together with the predicted subcellular locations of DmaHsp70s suggest that they likely fulfill distinct roles in cells. The transcription of four DmaHsp70s were changed during the recovery stage after thermal stress or oxidative stress. But the expression pattern of them were dissimilar. Collectively, these results collectively elucidated the identification of a previously uncharacterizedHsp70 lineage in animal and extended our understanding of the Hsp70 family.

Combined exposure to hypoxia and nanoplastics leads to negative synergistic oxidative stress-mediated effects in the water flea Daphnia magna

In this study, we investigated the combined effects of hypoxia and NPs on the water flea Daphnia magna, a keystone species in freshwater environments. To measure and understand the oxidative stress responses, we used acute toxicity tests, fluorescence microscopy, enzymatic assays, Western blot analyses, and Ingenuity Pathway Analysis. Our findings demonstrate that hypoxia and NPs exhibit a negative synergy that increases oxidative stress, as indicated by heightened levels of reactive oxygen species and antioxidant enzyme activity. These effects lead to more severe reproductive and growth impairments in D. magna compared to a single-stressor exposure. In this work, molecular investigations revealed complex pathway activations involving HIF-1α, NF-κB, and mitogen-activated protein kinase, illustrating the intricate molecular dynamics that can occur in combined stress conditions. The results underscore the amplified physiological impacts of combined environmental stressors and highlight the need for integrated strategies in the management of aquatic ecosystems.

Removal of Chromium (III) and Reduction in Toxicity in a Primary Tannery Effluent Using Two Floating Macrophytes.

Trivalent chromium (Cr(III)) is a contaminant with toxic activity. Its presence in waters and soils is usually related to industrial activities such as tanneries. The aim of this study was to compare the removal of Cr(III) in hydroponic solutions and tannery effluents using two floating macrophytes: Salvinia auriculata and Eichhornia crassipes. First, to determine the chromium removal capacity in solution and the bioaccumulation factor (BAF) in tissues of each plant, experiments were set up with contaminated solutions with Cr(III) concentrations of 2, 5, 10, 20, and 40 mg/L. Subsequently, both plant species were exposed to a primary tannery effluent contaminated with 12 mg/L of Cr(III) in order to study the removal capacity of organic and inorganic matter, as well as the acute toxicity in the water flea (Daphnia magna) and genotoxicity in zebrafish (Danio rerio). Tests carried out on nutrient solutions revealed that both plants have a high capacity for removing Cr(III) in solution. The BAF in tissues was higher in E. crassipes compared to S. auriculata. In the experiments with a tannery effluent, both species presented low nutrient and organic matter removal efficiency, but they showed good Cr(III) removal capacity, with average reduction values of 57% for S. auriculata and 54% for E. crassipes after 72 h of exposure. E. crassipes contributed most to the reduction in acute toxicity in D. magna, while S. auriculata did not show a similar effect. However, both plant species managed to reduce the genotoxicity marker in D. rerio when compared with the initial effluent and the control.

Regional and fine-scale local adaptation in salinity tolerance in Daphnia inhabiting contrasting clusters of inland saline waters.

Understanding the spatial scales at which organisms can adapt to strong natural and human-induced environmental gradients is important. Salinization is a key threat to biodiversity, ecosystem functioning and the provision of ecosystem services of freshwater systems. Clusters of naturally saline habitats represent ideal test cases to study the extent and scale of local adaptation to salinization. We studied local adaptation of the water flea Daphnia magna, a key component of pond food webs, to salinity in two contrasting landscapes-a dense cluster of sodic bomb crater ponds and a larger-scale cluster of soda pans. We show regional differentiation in salinity tolerance reflecting the higher salinity levels of soda pans versus bomb crater ponds. We found local adaptation to differences in salinity levels at the scale of tens of metres among bomb crater pond populations but not among geographically more distant soda pan populations. More saline bomb crater ponds showed an upward shift of the minimum salt tolerance observed across clones and a consequent gradual loss of less tolerant clones in a nested pattern. Our results show evolutionary adaptation to salinity gradients at different spatial scales, including fine-tuned local adaptation in neighbouring habitat patches in a natural landscape.

How Daphnia magna Defends Itself against Predators: Mechanisms and Adaptations in a Freshwater Microcosm

The freshwater water flea (Daphnia magna Straus, 1820) is prey for numerous predators. Yet it possesses a wide range of strategies to defend itself against predation. The aim of this work is to investigate the defensive mechanisms employed by D. magna to reduce predation by the coelenterate Hydra viridissima, and two planarians, Polycelis felina and Dugesia gonocephala. To do this, we used a freshwater microcosm. An additional aim is to investigate interactions with the presence of the isolated endosymbiotic algae from green hydra, thus combining and observing the interaction of the zooplankton and microalgal component. Each experiment included five replicates (13.5 °C, 25 °C), in crystallizing glass containers (60 mL volume, 60 mm diameter, 35 mm height), including satiated (fed with larvae of Artemia salina) and starved predators, respectively (one or five individuals of a particular predator species in one microcosm). As the isolated microalgae are unique, we tracked the following three mechanisms of Daphnia defense for the first time including precisely this microalgal component: (i) grouping (visual magnification), i.e., two or more Daphnia holding together; (ii) the phenomenon of overproduction, i.e., any number of Daphnia in one container above the 10 initially added individuals; and (iii) accelerated movement (“bullet movement”), i.e., high-speed movements in particular microcosms. The results provide new information for a better understanding of the interspecific relationships in systems that include both zooplankton and microalgal components.

Intraspecific and interspecific comparison of toxicity of ladybirds (Coleoptera: Coccinellidae) with contrasting colouration

Ladybirds (Coccinellidae) use toxic compounds, mostly alkaloids in their haemolymph, for defence against predators and other enemies. The toxicity of ladybirds to predators cannot be directly assessed because predators show avoidance reactions without ingesting the beetles. The alkaloid of ladybird Harmonia axyridis showed wide range toxicity to diverse non-target organisms. Thus, we used a quick, inexpensive and easy-to-perform method using bioassays on water flea Daphnia magna for comparative quantification of the toxicity (LD50) of whole body extracts from several species of ladybirds that differ in their warning colouration. Alien invasive aposematic polymorphic ladybird H. axyridis was more toxic than all the other species examined: aposematic Adalia bipunctata >cryptic Cynegetis impunctata >aposematic Coccinella septempunctata >slightly aposematic Calvia quatuordecimguttata. Three month old adults of H. axyridis were 3.8 times more toxic than two week and one month old adults. The two most common colour morphs (non-melanic novemdecimsignata and melanic spectabilis) did not differ in their toxicity. High toxicity of H. axyridis as compared to all other species examined may contribute to the invasiveness of this species.

From biomarkers to community composition: Negative effects of UV/chlorine-treated reclaimed urban wastewater on freshwater biota

The use of urban wastewater reclaimed water has recently increased across the globe to restore stream environmental flows and mitigate the effects of water scarcity. Reclaimed water is disinfected using different treatments, but their effects into the receiving rivers are little studied. Physiological bioassays and biomarkers can detect sub-lethal effects on target species, but do not provide information on changes in community structure. In contrast, official monitoring programs use community structure information but often at coarse taxonomic resolution level that may fail to detect species level impacts. Here, we combined commonly used biomonitoring approaches from organism physiology to community species composition to scan a broad range of effects of disinfection of reclaimed water by UV-light only and both UV/chlorine on the biota. We (1) performed bioassays in one laboratory species (water flea Daphnia magna) and measured biomarkers in two wild species (caddisfly Hydropsyche exocellata and the barbel Luciobarbus graellsii), (2) calculated standard indices of biotic quality (IBQ) for diatoms, benthic macroinvertebrates, and fishes, and (3) analysed community species composition of eukaryotes determined by Cytochrome Oxidase C subunit I (cox1) metabarcoding. Only the UV/chlorine treatment caused significant changes in feeding rates of D. magna and reduced antioxidant defenses, increased anaerobic metabolism and altered the levels of lipid peroxidiation in H. exocellata. However, inputs of reclaimed water were significantly associated with a greater proportion of circulating neutrophils and LG-PAS cells in L. graellsii. Despite IBQ did not discriminate between the two water treatments, metabarcoding data detected community composition changes upon exposure to UV/chlorine reclaimed water. Overall, despite the effects of UV/chlorine-treated water were transient, our study suggests that UV-light treated is less harmful for freshwater biota than UV/chlorine-treated reclaimed water, but those effects depend of the organizational level.