Natural products, specifically plant extracts with biological activity and the ability to act as botanical biopesticides are often mistakenly considered nontoxic. Scientific evidence indicates the contrary, and for this reason, the objective of this work was to evaluate the toxicity of extracts obtained from Petiveria alliacea L. (Caryophyllales, Phytolaccaceae) using Daphnia magna Straus (Cladocera, Daphniidae) as a bioindicator to identify the plant extracts and the respective concentrations that present the highest toxicity. Leaves of P. alliacea were collected in the Peruvian amazone. From this material, three types of extract (hexane, ethanolic and aqueous) were prepared, which were used in the bioassays with D. magna to find the least toxic extract. Acute toxicity bioassays with D. magna during 48 h of exposure to hexane, ethanolic, and aqueous extracts yielded median lethal concentration (LC50) values of 26.9, 230.6, and 657.9 mg L-1, respectively. The aqueous extract presented the lowest toxicity, causing minimal D. magna mortality in the range of 6.67 to 13.33% at concentrations of 10 and 100 mg L-1. This result enables the efficient use of this plant species in a sustainable manner with a minimal environmental impact for the future development of natural products for pest control.

Juvenile hormone (JH), together with ecdysone, regulates molting, metamorphosis, growth, and reproduction in arthropods. The effects of its analogs used as insecticides on nontarget species are of concern. Since JH and JH analogs (JHAs) induce male offspring in daphnids, which generally reproduce by parthenogenesis, short-term JH activity screening assay (JHASA) using the male offspring ratio as an endpoint has been developed as a detection method for JHA. However, the production of male offspring is also induced by environmental stresses such as temperature, short-day length, overcrowding, and food limitation. Thus, it is vital to prevent non-chemical stresses from inducing male offspring during the test to detect chemicals with potential JH activity accurately. Therefore, we investigated the effects of temperature (low and high), hardness, high density with low feeding, and day length on male production utilizing JHASA. Male offspring were not strongly induced by any stresses in JHASA, although the male ratios of 4-12% were observed in the preculture under high density (≥70 daphnid/L) and constant darkness. The Clone A strain was relatively more sensitive to high density and day length compared with the strain from National Institute for Environmental Studies (NIES). The selection of strains that rarely produce males under non-chemical stresses and finding the culturing conditions for each strain appropriate for not-inducing male offspring are recommended to control and prevent male offspring induction during JHASA.

Microplastic risk assessment often characterizes primary plastics, plastics intentionally manufactured at the micro- and nanoscale, or plastics collected within the natural environment, which often lack repeatability and the volume necessary for reliable hazard characterization. There is limited understanding of how environmentally representative plastics prepared at the microscale impact aquatic organisms. The aim of the present study was to create environmentally representative microplastics and characterize their toxicokinetics and hazards. Plastic cups and forks were micronized to <120 μm particles and Daphnia magna were exposed for 48 h at concentrations ranging from 0.01 mg/L to 100 mg/L. Uptake and depuration experiments were conducted at the highest concentration where accumulation was confirmed. Raman spectroscopy identified that both plastics were polystyrene and had similar size distributions. Microplastics were not acutely toxic but accumulated and rapidly depurated. Toxicokinetics demonstrated that cup MPs were consumed at higher rates than fork MPs despite similar physical characteristics. Daphnia magna preferentially selected smaller particles from the heterogenous suspensions. Future research will need to further explore the relationship between physicochemical properties, particularly size, and ecotoxicity. The study focused on mortality as the primary hazard endpoint. However alternative, sublethal biomarkers may be more appropriate in describing the effects of microplastic exposure.

In the frame of bioeconomy and circular economy, wastewater sludge (WS) could be a good candidate for its use in agriculture as fertilizer, due to its high content of organic matter, N and P, but on the other hand, it is full of toxicants such as heavy metal, microplastics, detergent, antibiotics, and so on that can reach groundwater and water bodies in leachate form. In this study, we have investigated different sludge concentrations in the eluate form, combined and not with PVC on two different freshwater organisms Selenastrum capricornutum and Daphnia magna, using ecotoxicity tests. At the endpoint, we have evaluated inhibition growth rate, oxidative stress, and pigments production for S. capricornutum, while in case of D. magna, we have assessed organism immobilization and development. From our results, it emerged that at the higher WS concentration, there was not inhibition growth rate, while at oxidative stress, it was higher in algae treated with WS and PVC. Higher Chl-a production was shown for algae treated with 0.3 g/L of sludge coupled with PVC, where higher phaeopigments production were recorded for algae treated with 0.3 g/L of WS. D. magna has shown an opposite trend when compared with algae, where at the highest WS concentrations supplied was corresponding to an increased mortality explaned as the highest immobility percentage. PRACTITIONER POINTS: Wastewater sludge is used in agriculture as fertilizer. PVC microplastic presence and associate ecotoxicity was tested. PVC presence increased oxidative stress in S. capricornutum. D. magna was significantly affected by sludge concentrations supplied.

Suspended particulate matter (SPM) and biofilm are critical in removing contaminants in aquatic environments, but the environmental behavior and ecological toxicity of SPM-biofilm aggregates modulated by turbulence intensities are largely unknown. This study determined the removal pathways of microcystin-LR (MC-LR) by SPM and its biofilm under different turbulence intensities (2.25 × 10-3, 1.01 × 10-2, and 1.80 × 10-2 m2/s3). Then, we evaluated the toxicity of SPM-biofilm aggregates to Daphnia magna. The results revealed that SPM contributed to the adsorption of MC-LR, and the removal of MC-LR can be accelerated with biofilm formation on SPM, with 95.66 % to 97.45 % reduction in MC-LR concentration under the studied turbulence intensities. Higher turbulence intensity triggered more frequent contact of SPM and MC-LR, formed compact but smaller clusters of SPM-biofilm aggregates, and enhanced the abundance of mlrA and mlrB; thus benefiting the adsorption, biosorption, and biodegradation of MC-LR. Furthermore, the SPM-biofilm aggregates formed in turbulent water triggered oxidative stress to Daphnia magna, while a weak lethal toxic effect was identified under moderate turbulence intensity. The results indicate that the toxicity of SPM-biofilm aggregates fail to display a linear relationship with turbulence intensity. These findings offer new perspectives on understanding the environmental behavior and ecological outcomes of SPM and its biofilms in turbulent aquatic environments.

Polycyclic aromatic hydrocarbons (PAHs) arise from incomplete combustion of oil, coal, and gasoline, with lipophilic properties facilitating their widespread distribution and persistence. Due to their biochemical attributes, PAHs can accumulate in animal tissues, potentially causing mutagenic and carcinogenic effects. Since the industrial revolution, PAH concentrations in the environment have risen, with lakes showing levels from 0.159 to 33,090 μg/kg sediment. Despite acute toxicity studies showing adverse effects on freshwater organisms, the long-term impacts and synergistic interactions with other pollutants remain largely unexplored. This study investigates the impact of phenanthrene (PHE), a prominent PAH found in aquatic environments, on Daphnia magna, a species of significant ecological importance in freshwater ecosystems globally, being both a sentinel species for chemical pollution and a keystone organism in freshwater aquatic ecosystems. Leveraging the dormancy of D. magna, which spans decades or even centuries, we exposed strains with diverse histories of chemical contaminant exposure to environmentally relevant concentrations of PHE. Initially, acute exposure experiments were conducted in accordance with OECD guidelines across 16 Daphnia strains, revealing substantial variation in acute toxic responses, with strains naïve to chemical pollutants showing the lowest toxicity. Utilizing the median effect concentration EC10 derived from acute exposures, we assessed the impacts of chronic PHE exposure on life history traits and ecological endpoints of the 16 strains. To elucidate how historical exposure to other environmental stressors may modulate the toxicity of PHE, temporal populations of D. magna resurrected from a lake with a well-documented century-spanning history of environmental impact were utilized. Our findings demonstrate that PHE exposure induces developmental failure, delays sexual maturation, and reduces adult size in Daphnia. Populations of Daphnia historically exposed to chemical stress exhibited significantly greater fitness impacts compared to naïve populations. This study provides crucial insights into the augmented effects of PAHs interacting with other environmental stressors.

Agri-chemicals such as fungicides are applied in natural settings and hence are exposed to the environment\'s ultraviolet (UV) light. Recently, many fungicides in commerce are being modified as nano-enabled formulations to increase agricultural productivity and reduce potential off-target effects. The present study investigated the impacts of sunlight-grade UV emission on the effects of either conventional or nano-enabled azoxystrobin (Az or nAz, respectively), a commonly applied agricultural fungicide, on Daphnia magna. Daphnids were exposed to increasing concentrations of Az or nAz under either full-spectrum (Vis) or full-spectrum Vis + UV (Vis + UV) lighting regimes to evaluate LC50s. Az LC50 was calculated at 268.8 and 234.2 μg/L in Vis or Vis + UV, respectively, while LC50 for nAz was 485.6 and 431.0 μg/L under Vis or Vis + UV light, respectively. Daphnids were exposed to 10% LC50 of either Az or nAz under Vis or Vis + UV lighting regime for 48 h or 21 d (acute and chronic, respectively). By 48 h, both Az and nAz reduced O2 consumption and increased TBARS. Heart rate was increased in Az-exposed daphnids but not in nAz groups. Neither of the two chemicals impacted thoracic limb activity. In 21 d exposures, Az significantly reduced biomass production and fecundity, but nAz groups were not significantly different from controls. The results of the present study demonstrate that conventional Az is more toxic to D. magna at lethal and sub-lethal levels in acute and chronic exposures, and sunlight strength UV can potentiate both acute and chronic effects of Az and nAz on D. magna.

Perfluoroalkyl substances (PFAS) are common environmental pollutants, but their toxicity framework remains elusive. This research focused on ten PFAS, evaluating their impacts on two ecotoxicologically relevant model organisms from distinct trophic levels: the crustacean Daphnia magna and the unicellular green alga Raphidocelis subcapitata. The results showed a greater sensitivity of R. subcapitata compared to D. magna. However, a 10-day follow-up to the 48 h immobilisation test in D. magna showed delayed mortality, underlining the limitations of relying on EC50 s from standard acute toxicity tests. Among the compounds scrutinized, Perfluorodecanoic acid (PFDA) was the most toxic to R. subcapitata, succeeded by Perfluorooctane sulfonate (PFOS), Perfluorobutanoic acid (PFBA), and Perfluorononanoic acid (PFNA), with the latter being the only one to show an algicidal effect. In the same species, assessment of binary mixtures of the compounds that demonstrated high toxicity in the single evaluation revealed either additive or antagonistic interactions. Remarkably, with an EC50 of 31 mg L-1, the short-chain compound PFBA, tested individually, exhibited toxicity levels akin to the notorious long-chain PFOS, and its harm to freshwater ecosystems cannot be ruled out. Despite mounting toxicological evidence and escalating environmental concentrations, PFBA has received little scientific attention and regulatory stewardship. It is strongly advisable that regulators re-evaluate its use to mitigate potential risks to the environmental and human health.

Cyanobacterial blooms, resulting from serious eutrophication, can produce various cyanotoxins and severely disrupt aquatic ecosystems. Inducible defenses are adaptive traits developed by prey in response to predation risks. However, the effects of the increasing proportion of cyanobacteria and cyanotoxins produced during cyanobacterial blooms on the inducible defenses of cladocerans, particularly in terms of behavioral defenses, remain unclear. In this study, we selected Daphnia magna and investigated the defensive traits against predation risks by the predator Rhodeus ocellatus under different ratios of cyanobacteria (Dolichospermum flos-aquae) and green algae (Scenedesmus obliquus), as well as varying concentrations of anatoxin-a (ATX), a cyanotoxin. We recorded the inducible defensive traits involving to morphology, behavior, and offspring production of D. magna. Results showed that the body length of D. magna at sexual maturity and the number of offspring in the first brood were significantly reduced by the presence of D. flos-aquae. Moreover, when the proportion of D. flos-aquae reached 75% and 100%, D. magna did not develop to sexual maturity. Furthermore, D. flos-aquae inhibited the formation of inducible behavioral defense of D. magna, with a stronger inhibitory effect as the proportion of D. flos-aquae increased. In this experiment, the effects of ATX on the morphological traits at sexual maturity and offspring production of D. magna were minor, but ATX still had the potential to inhibit the formation of inducible behavioral defense. We confirmed that changes in the proportion of cyanobacteria and green algae as well as the production of ATX by cyanobacteria during cyanobacterial blooms can affect the growth, development, and inducible defensive traits of cladocerans, potentially altering their population dynamics during such events.

Zeolite type 5A combined with the magnetic properties of maghemite nanoparticles facilitate the rapid absorption of heavy metals, which makes them an interesting proposal for the remediation of water contaminated with lead and arsenic. However, the physicochemical analysis related to concentration and size for the use of this magnetic zeolite composite (MZ0) in water bodies and the possible toxicological effects on aquatic fauna has not yet been carried out. The main objective of the research work is to determine lethal concentrations that cause damage to Daphnia magna based on LC50 tests, morphology, reproductive rate, and quantification of the expression of three genes closely involved in the morphological development of vital structures (Glass, NinaE, Pph13). To achieve this objective, populations of neonates and young individuals were used, and results showed that the LC50 for neonates was 11,314 mg L-1, while for young individuals, it was 0.0310 mg L-1. Damage to morphological development was evidenced by a decrease in eye size in neonates, an increase in eye size in young individuals, variations in the size of the caudal spine for both age groups, and slight increases in the heart size, body, and antenna for both age groups. The reproductive rate of neonates was not affected by the lower concentrations of MZ0, while in young individuals, the reproductive rate decreased by more than 50% from the minimum exposure concentration of MZ0. And for both ages, Glass gene expression levels decreased as the MZ0 concentration increased. Also, the MZ0 evidenced its affinity for the exoskeleton of D. magna, which was observed using both light microscopy and electron microscopy. It is concluded that MZ0 did not generate significant damage in the mortality, morphology, reproductive rate, or gene expression in D. magna at lower concentrations, demonstrating the importance of evaluating the possible impacts on different life stages of the cladoceran.