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In the packaging materials sector, increasing globalization has created the need for increased efforts to develop consumer protection measures. Consequently, new packaging materials are being sought to replace petroleum-based materials in the future. For this reason, global awareness of the environmental problems associated with the use of synthetic and non-degradable packaging has increased the attention paid to bio-packaging based on natural and biodegradable polymers. The bio-packaging sector is developing innovations to address the sustainability issues facing the food packaging industry. Our research has shown that green matcha extract can be a promising source of antioxidants for the production of bioactive pectin films. This study further confirmed that green matcha extract can be a promising source of antioxidants for the production of bioactive pectin films. The antioxidant activity test showed high activity of films containing matcha extract. The antioxidant activity of films without matcha addition, P, PJ, PC, PJC, was negligible. The addition of matcha to the polymer matrix did not significantly affect the mechanical properties (TS, EB) of the films obtained. The addition of cellulose had the greatest effect on changing the mechanical properties. It caused a twofold increase in the mechanical properties of the obtained packaging films. The addition of matcha significantly improved the barrier properties (for PM films, the WVTR was 3.40 [g/m2d]; for PJM films the WVTR was 1.70 [g/m2d]). The green packaging films showed no toxic effects on the plants (Phacelia tanacetifolia, Salvia hispanica, Brassica napus) and invertebrates (Daphnia pulex, Chaoborus, Chironomus aprilinus) tested. The half-solubility time of the membranes in a solution mimicking gastric acid was also determined. The longest half-dissolution time of the films was about 2 min. Our research has therefore shown that the biodegradable and environmentally safe green packaging films with antioxidant activity that we have developed can be used as edible functional casings in the future, e.g., for sausages and other food products. [ABSTRACT FROM AUTHOR]

The on‐going anthropogenic degradation of freshwater habitats has drastically altered the environmental supply of both nutrients and common pollutants. Most organisms living in these altered habitats experience interactive effects of various stressors that can initiate adjustments at multiple levels impacting their fitness. Hence, studies measuring response to a single environmental parameter fail to capture the complexities of the status quo. We tested both the individual and the interactive effect of arsenic (As) exposure, food quantity, and dietary phosphorus (P)‐supply on six life‐history traits (Juvenile Growth Rate; Adult Growth Rate; Age and Size at Maturity, Lifespan, and Fecundity) as surrogates for organismal fitness in the keystone aquatic grazer Daphnia pulex. We also tested the effect of food quantity and P‐supply on somatic As accumulation in Daphnia. Our results indicated an influence of P‐supply on neonatal growth and an influence of As and food quantity on growth and maintenance later in life. Maturation was strongly influenced by all three variables, with no reproduction observed in the presence of two or more environmental stressors. We found a strong interaction between As and dietary P, with increased P‐supply intensifing the toxicity effect of As. No such effects were seen between As and food quantity, indicating a differential role of quantity versus quality on As toxicity. We found a nominal effect of diet on somatic As accumulation. The results from the present study emphasize the importance of considering such interactions between co‐occurring environmental stressors and the dietary status of organisms, to better predict and manage impacts and risks associated with common environmental toxicants in highly vulnerable ecosystems. Environ Toxicol Chem 2024;43:1807–1819. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. [ABSTRACT FROM AUTHOR]

Despite evolutionary biology's obsession with natural selection, few studies have evaluated multigenerational series of patterns of selection on a genome-wide scale in natural populations. Here, we report on a 10-y population-genomic survey of the microcrustacean Daphnia pulex. The genome sequences of >800 isolates provide insights into patterns of selection that cannot be obtained from long-term molecularevolution studies, including the following: the pervasiveness of near quasi-neutrality across the genome (mean net selection coefficients near zero, but with significant temporal variance about the mean, and little evidence of positive covariance of selection across time intervals); the preponderance of weak positive selection operating on minor alleles; and a genome-wide distribution of numerous small linkage islands of observable selection influencing levels of nucleotide diversity. These results suggest that interannual fluctuating selection is a major determinant of standing levels of variation in natural populations, challenge the conventional paradigm for interpreting patterns of nucleotide diversity and divergence, and motivate the need for the further development of theoretical expressions for the interpretation of population-genomic data. [ABSTRACT FROM AUTHOR]

Ribosomal DNA (rDNA) has a vital role in ribosome biogenesis as it contains the genes that encode ribosomal RNA (rRNA) separated by intergenic spacers (IGSs). The rRNA genes occur in hundreds to tens of thousands of copies per haploid genome in eukaryotes and are generally highly conserved with low variation within species. Due to the repetitive nature and large size of rDNA arrays, detecting intraindividual variation can be difficult. In this study, we use whole-genome sequences of 169 Daphnia pulex individuals from 10 natural populations to measure the copy number and sequence variation in rDNA. This revealed that variation in rDNA copy number between individuals spans an order of magnitude. We further observed a substantial level of sequence variation within individual genomes. As expected, single-nucleotide polymorphisms occurred in regions of lower functional constraint such as the IGS and expansion segments of the rRNA genes. The presence of strong linkage disequilibrium among variants facilitated identification of haplotypes within each population. Although there was evidence of recombination among haplotypes from different populations, it is insufficient to eliminate linkage disequilibrium within populations. Estimating copy number and haplotype diversity within individuals revealed that the level of intraindividual sequence variation is not strongly correlated with copy number. The observed patterns of variation highlight a complex evolutionary history of rDNA in D. pulex. Future research should explore the functional implications of rDNA copy number and sequence variation on organismal phenotypes. [ABSTRACT FROM AUTHOR]

Evolutionary theory suggests the hypothesis that among genetically isolated populations, phenotypic variation should be smaller in populations with smaller ratios of nonsynonymous to synonymous substitutions (dN/dS) because smaller dN/dS ratios occur under greater purification selections. Two distinct lineages (JPN1 and JPN2) of panarctic Daphnia cf. pulex sensu Hebert (1995), invaded independently into Japan from North America, provide an excellent opportunity to test this hypothesis because the earlier invader JPN1 has a lower dN/dS ratio than JPN2. Therefore, we examined phenotypic variations in fitness‐related traits, including digestive, life‐history, and morphological traits, among several genotypes within these two lineages. We found that phenotypic variations were smaller within the JPN1 lineage than within the JPN2 lineage. Furthermore, within‐lineage variation of the phenotypic plasticity to changing food levels was smaller in the JPN1 lineage than in the JPN2 lineage. These results support the hypothesis that the JPN1 lineage has been more efficiently subjected to negative selection. However, the magnitude of the phenotypic plasticity of these traits was, on average, at the same level between the JPN1 and JPN2 lineages and its direction differed among genotypes of these lineages, suggesting that the JPN2 genotypes might have exploited niches that were different from those of the JPN1 genotypes. [ABSTRACT FROM AUTHOR]

Pit lakes are currently being investigated as a way to store and reclaim waste materials in the Alberta Oil Sands (AOS) region, Canada. Lake Miwasin (LM) is a pilot-scale pit lake consisting of treated fine tailings overlayed with oil sands process-affected water (OSPW) blended with fresh surface water. In October 2021, the surface water contained a mean concentration of 1.33 ± 0.04 µg/L dissolved selenium (Se), slightly above the Canadian Council of Ministers of Environment water quality guideline for long-term protection of aquatic life (1 µg Se/L). This study assessed the bioaccumulation of Se by the cladoceran Daphnia pulex under laboratory conditions through both aqueous and dietary exposure routes for comparison to field-collected specimens. In 12-day semi-static tests, lab-cultured D.pulex were exposed to water, and algae grown in media spiked with selenate. Results showed that Se bioaccumulation by lab-cultured D. pulex increased in all exposure treatments from days 5 to 12, with maximum Se concentrations of 3.08–3.47 µg/g dry weight (dw) observed within the exposure range tested. Interestingly, lower Se bioaccumulation concentrations (1.26–1.58 µg/g dw) were observed in the highest dissolved Se and dietary Se treatments, suggesting potential internal regulatory mechanisms. In addition, native D. pulex (LM) collected from Lake Miwasin and cultured in-house were exposed in 8-day semi-static tests to Lake Miwasin surface water and algae cultured in Lake Miwasin surface water. Selenium bioaccumulation in native D. pulex (LM) ranged from 2.00 to 2.04 µg/g dw at day 8 and was not significantly different (p > 0.05) compared to Se concentrations in D. pulex collected from Lake Miwasin (2.15 ± 0.28 µg/g) in summer 2022. [ABSTRACT FROM AUTHOR]

Daphnia can avoid predation by sensing fish kairomones and producing inducible defenses by altering the phenotype. In this study, the results showed that the morphological and life‐history strategies of two Daphnia species (Daphnia pulex and Daphnia sinensis) exposed to Aristichthys nobilis kairomones. In the presence of fish kairomones, the two Daphnia species exhibited significantly smaller body length at maturity, smaller body length of offspring at the 10th instar, and longer relative tail spine of offspring. Nevertheless, other morphological and life‐history traits of the two Daphnia species differed. D. pulex showed a significantly longer relative tail spine length and earlier age at maturity after exposure to fish kairomones. The total offspring number of D. sinensis exposed to fish kairomones was significantly higher than that of the control group, whereas that of D. pulex was significantly lower. These results suggest that the two Daphnia species have different inducible defense strategies (e.g., morphological and life‐history traits) during prolonged exposure to A. nobilis kairomones, and their offspring also develop morphological defenses to avoid predation. It will provide reference for further exploring the adaptive evolution of Daphnia morphology and life‐history traits in the presence of planktivorous fish. [ABSTRACT FROM AUTHOR]

Road salt (NaCl) is the most common deicer used to melt snow and ice from roadways in cold climates, but its use results in runoff of sodium and chloride ions into sewers and into soil and freshwater ecosystems. Road salt causes harmful effects on wetland habitats, which host an abundance of macroinvertebrate species. Alternatives to road salt, such as beet juice, are commercially available, but their impact on invertebrates is less well studied. We examined the impact of road salt alternatives on four invertebrate species (Daphnia pulex, Eisenia fetida, Heterocypris sp., and Culicoides sp. larvae), some of which come from a naturally brackish area. Our expectation was that road salt alternatives (beet salt, molasses, pickle juice, and sand) would be less toxic to the animals than traditional road salt. This study comprised two experiments: the first exposed individuals of the four invertebrate species to road salt alternatives, and the second exposed the invertebrates to a range of road salt concentrations. Only individuals of D. pulex were negatively impacted by road salt. The other invertebrates were not significantly affected by any salt alternative, demonstrating interspecific differences in response to salt. Species found in naturally salty environments were more tolerant of both road salt and alternatives. This research shows species‐ and habitat‐specific responses to road salt alternatives, which has implications for municipalities looking to salt roads without negatively impacting freshwater ecosystems. [ABSTRACT FROM AUTHOR]

Octahydro‐tetramethyl‐naphthalenyl‐ethanone (OTNE) is a high–production volume fragrance material used in various down‐the‐drain consumer products. To assess aquatic risk, the Research Institute for Fragrance Materials (RIFM) uses a tiered data‐driven framework to determine a risk characterization ratio, where the ratio of the predicted‐environmental concentration to the predicted‐no‐effect concentration (PNEC) of <1 indicates an acceptable level of risk. Owing to its high production volume and the conservative nature of the RIFM framework, RIFM identified the need to utilize a species sensitivity distribution (SSD) approach to reduce the PNEC uncertainty for OTNE. Adding to the existing Daphnia magna, Danio rerio, and Desmodesmus subspicatus chronic studies, eight new chronic toxicity studies were conducted on the following species: Navicula pelliculosa, Chironomus riparius, Lemna gibba, Ceriodaphnia dubia, Hyalella azteca, Pimephales promelas, Anabaena flos‐aquae, and Daphnia pulex. All toxicity data were summarized as chronic 10% effect concentration estimates using the most sensitive biological response. Daphnia magna was the most sensitive (0.032 mg/L), and D. subspicatus was the least sensitive (>2.6 mg/L, the OTNE solubility limit). The 5th percentile hazardous concentration (HC5) derived from the cumulative probability distribution of the chronic toxicity values for the 11 species was determined to be 0.0498 mg/L (95% confidence interval 0.0097–0.1159 mg/L). A series of "leave‐one‐out" and "add‐one‐in" simulations indicated the SSD was stable and robust. Add‐one‐in simulations determined that the probability of finding a species sensitive enough to lower the HC5 two‐ or threefold was 1/504 and 1/15,300, respectively. Given the high statistical confidence in this robust SSD, an additional application factor protection is likely not necessary. Nevertheless, to further ensure the protection of the environment, an application factor of 2 to the HC5, resulting in a PNEC of 0.0249 mg/L, is recommended. When combined with environmental exposure information, the overall hazard assessment is suitable for a probabilistic environmental risk assessment. Environ Toxicol Chem 2024;43:1378–1389. © 2024 SETAC [ABSTRACT FROM AUTHOR]

Statistical models used in toxicity experiments have been quite useful tools in interpreting the organism's susceptibility, exposure response, amount of tolerable concentration, and function of tolerance time. In order to determine and evaluate the toxic effects of boron on Daphnia pulex (Leydig, 1860), different boron concentrations considered to be tolerable in aquatic ecosystems were tested for D. pulex. Percentage mortality rates at different boron concentrations and probit regression estimates at these concentrations were investigated through the static method. Probit analysis in this study revealed that rising boron concentrations led to mortality, and that finding was statistically significant with P value. These results indicate that the use of episodic boron, which enters the aquatic ecosystem through natural or unnatural means, should be planned due to its potential for stress or toxicity situations on organisms. [ABSTRACT FROM AUTHOR]

Macrophytes and cladocerans represent the main antagonistic groups that regulate phytoplankton biomass; however, the mechanism behind this interaction is unclear. In laboratory conditions, we separately evaluated the effects of three submerged macrophytes (Ceratophyllum demersum, Myriophyllum aquaticum, and Stuckenia pectinata), as well as their exudates, and plant-associated microbiota (POM < 25 µm) + exudates on the population growth of Daphnia cf. pulex and Simocephalus cf. mixtus. Living Ceratophyllum, exudates, and POM < 25 µm + exudates exhibited the most robust positive effects on Simocephalus density and the rate of population increase (r). Subsequently, we examined the effects of Ceratophyllum on the filtration and feeding rates of Simocephalus and Daphnia, revealing significant (p < 0.001) promotion of filtration and feeding in Simocephalus but not in Daphnia. To elucidate the specific effects of this macrophyte on Simocephalus demography, we assessed selected life table variables across the same treatments. The treatments involving exudates and living Ceratophyllum resulted in approximately 40% longer survivorship and significantly (p < 0.01) enhanced fecundity. Our findings indicate that exudates from submerged macrophytes positively influence Simocephalus demography by increasing filtration rates, survivorship, and fecundity. This synergy suggests a substantial impact on phytoplankton abundance. [ABSTRACT FROM AUTHOR]

Asexual organisms are ubiquitous. While being members of a single species with the same ecological requirements, multiple asexual genotypes within a species are often found in a single habitat. Niche partitioning in time and space among genotypes has been proposed to explain this phenomenon. However, it is not clear whether these different genotypes co‐occur in the long term.Therefore, we examined the population dynamics of two asexual Daphnia cf. pulex genotypes (JPN1 and JPN2) over 9 years in a small mountain lake. These two genotypes consistently occurred in the same seasons and layers, suggesting that niche partitioning cannot explain their long‐term co‐occurrence. The abundance of JPN1 was typically higher in most years. However, the abundance of dormant eggs in lake sediments was at the same level between the genotypes.Results of a laboratory experiment showed that JPN1 competitively excluded JPN2. However, many JPN2 individuals produced dormant eggs before JPN1 competitively excluded them from the experiment. Furthermore, the competitively inferior JPN2 produced dormant eggs abundantly in the medium containing a crowding cue from JPN1 while no such trend was observed for JPN1.These results showed that a competitively inferior asexual genotype could maintain a population with a competitively superior genotype for a long period of time because it had the ability to detect an increase in competitors and produce dormant eggs each year before being competitively eliminated.Based on these results, we suggest that the variation in genotype‐specific response in dormant egg production to environmental change plays a key role in the long‐term co‐occurrence of different asexual genotypes in single habitats. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

Prymnesium parvum is a harmful alga found in brackish waters worldwide whose toxins can be lethal to aquatic organisms. Established field methods to control blooms of this species, however, are unavailable. Earlier studies showed that various extracts of giant reed (Arundo donax) can suppress P. parvum growth and that ellipticine, an allelochemical present in giant reed, is a potent algicide against this species. The unintended effects of giant reed products on nontarget organisms, however, are not fully understood. This study determined the effects of giant reed leachate (aqueous extract of dried chips) and ellipticine on growth of P. parvum and the green microalga Chlorella sorokiniana; survival and reproduction of the planktonic crustacean Daphnia pulex; and hatching success, larval survival, and larval swimming behavior of the teleost fish Danio rerio. Leachate made with 3 g chips L−1 was lethally toxic to P. parvum and D. pulex, stimulated C. sorokiniana growth, and impaired D. rerio behavior. Leachate at 1 g L−1 fully suppressed P. parvum growth, had moderate effects on D. pulex reproductive output, and had no effects on D. rerio. Ellipticine at 0.01 mg L−1 irreversibly inhibited P. parvum growth, acutely but reversibly inhibited C. sorokiniana growth, slightly delayed D. pulex reproduction, and had no effects on D. rerio. These observations suggest that when applied at appropriate concentrations, natural products derived from giant reed can be used as tools to specifically control P. parvum growth with minimal effects on nontarget species. [ABSTRACT FROM AUTHOR]

The timing and duration of dormancy can be influenced by many environmental cues, such as changes in light and temperature. This study examined the early termination of dormancy in the absence of any major changes in environmental cues. We examined maternal influence on the early termination of dormancy in Daphnia pulex, as well as the effects of early termination of dormancy on life-history traits. In a mesocosm experiment, we found that a substantial proportion of individuals emerged early, prior to experiencing a cold shock, with no significant change in environmental conditions from the maternal environment. We found that siblings from the same ephippium were more likely to emerge at the same time, even after dissection and separation, suggestive of a maternal effect. Additionally, we found that the time to first reproduction was significantly delayed in individuals that emerged early, and that early hatchers also produced fewer dormant embryos in subsequent asexual generations. We suggest that early dormancy termination may be an adaptive strategy influenced by maternal effects to generate variation in developmental timing when the length and harshness of the unfavorable season is unpredictable. [ABSTRACT FROM AUTHOR]

The high-elevation plateaus of the inter-Andean valleys are home to shallow lakes that have become eutrophic. These lakes share similarities with shallow lakes in temperate and subtropical areas. Because native species diversity is low, invasive species dominate the fish and macrophytes communities. The study aimed to investigate the behavioral response of the local Daphnia pulex from the Andean shallow Lake Yahuarcocha to the exotic submerged macrophyte Egeria densa and the exotic fish Poecilia reticulata. Laboratory habitat choice experiments revealed that D. pulex from Lake Yahuarcocha strongly avoid E. densa, irrespective of the presence of the fish P. reticulata or chemical cues indicating fish predation on D. pulex. This observation could be explained by the fact that P. reticulata displayed a strong attraction to E. densa during the daytime, probably to avoid bird predation. D. pulex from the nearby Lake San Pablo where P. reticulata is absent but where the fish community is dominated by Oncorhynchus mykiss displayed the same avoidance behavior to submerged macrophytes as D. pulex from Lake Yahuarcocha. These results indicate that macrophytes in these high-elevation shallow lakes may not facilitate top-down control of phytoplankton, since plants do not offer refuge to D. pulex from fish predation. [ABSTRACT FROM AUTHOR]

Previous research has shown that elevated temperature and environmental hypoxia are associated with a reduced representation of larger‐bodied species in communities of ectotherms, suggesting their competitive disadvantage resulting from increased oxygen demands and limited oxygen availability. To investigate this, we conducted a comparative analysis of two pairs of competing zooplankton species: Daphnia galeata (smaller‐bodied) and Daphnia pulex (larger‐bodied), as well as D. pulex (smaller‐bodied) and Daphnia magna (larger‐bodied). We monitored their population dynamics under different oxygen concentrations and temperatures, and additionally measured hemoglobin concentration in the latter pair. Contrary to expectations, the results demonstrated that the larger‐bodied species outcompeted the smaller‐bodied ones in all experimental treatments involving the first pair of species. In the second pair, elevated temperature favored the smaller‐bodied species, while environmental hypoxia had the opposite effect. The competitive advantage of D. pulex under elevated temperature likely stemmed from their ability to allocate energy from somatic growth to reproduction, as indicated by the increased number of reproducing females. Conversely, the competitive advantage of D. magna under environmental hypoxia may be attributed to their ability to reallocate energy from reproduction to somatic growth, as indicated by lower reproductive effort and greater plasticity in hemoglobin production. These findings indicate that elevated temperature and hypoxia can independently and jointly influence interspecific competitive abilities by impacting the pace of life. However, the numerical dominance of smaller‐sized species in warm and hypoxic zooplankton communities cannot be explained by their enhanced competitive abilities resulting from a more favorable oxygen supply demand balance. [ABSTRACT FROM AUTHOR]

Understanding mechanisms that promote the maintenance of biodiversity (genetic and species diversity) has been a central topic in evolution and ecology. Previous studies have revealed that diapause can contribute to coexistence of competing genotypes or species in fluctuating environments via the storage effect. However, they tended to focus on differences in reproductive success (e.g. seed yield) and diapause termination (e.g. germination) timing. Here we tested whether different photoperiodic responses in diapause induction can promote coexistence of two parthenogenetic (asexual) genotypes of Daphnia pulex in Lake Fukami-ike, Japan. Through laboratory experiments, we confirmed that short day length and low food availability induced the production of diapausing eggs. Furthermore, we found that one genotype tended to produce diapausing eggs in broader environmental conditions than the other. Terminating parthenogenetic reproduction earlier decreases total clonal production, but the early diapausing genotype becomes advantageous by assuring reproduction in 'short' years where winter arrival is earlier than usual. Empirically parameterized theoretical analyses suggested that different photoperiodic responses can promote coexistence via the storage effect with fluctuations of the growing season length. Therefore, timing of diapause induction may be as important as diapause termination timing for promoting the maintenance of genetic diversity in fluctuating environments. [ABSTRACT FROM AUTHOR]

All animals and plants respond to changes in the environment during their life cycle. This flexibility is known as phenotypic plasticity and allows organisms to cope with variable environments. A common source of environmental variation is predation risk, which describes the likelihood of being attacked and killed by a predator. Some species can respond to the level of predation risk by producing morphological defences against predation. A classic example is the production of so‐called 'neckteeth' in the water flea, Daphnia pulex, which defend against predation from Chaoborus midge larvae. Previous studies of this defence have focussed on changes in pedestal size and the number of spikes along a gradient of predation risk. Although these studies have provided a model for continuous phenotypic plasticity, they do not capture the whole‐organism shape response to predation risk. In contrast, studies in fish and amphibians focus on shape as a complex, multi‐faceted trait made up of different variables. In this study, we analyse how multiple aspects of shape change in D. pulex along a gradient of predation risk from Chaoborus flavicans. These changes are dominated by the neckteeth defence, but there are also changes in the size and shape of the head and the body. We detected change in specific modules of the body plan and a level of integration among modules. These results are indicative of a complex, multi‐faceted response to predation and provide insight into how predation risk drives variation in shape and size at the level of the whole organism. [ABSTRACT FROM AUTHOR]

Mechanistic effect modeling is a promising tool to improve the ecological realism of environmental risk assessment. An open question for the mechanistic modeling of metal toxicity is whether the same physiological mode of action (PMoA) could be assumed for closely related species. The implications of various modeling choices, such as the use of parameter point estimates and assumption of simplistic toxicodynamic models, are largely unexplored. We conducted life‐table experiments with Daphnia longispina, Daphnia magna, and Daphnia pulex exposed to the single metals Cu, Ni, and Zn, and calibrated toxicokinetic–toxicodynamic (TKTD) models based on dynamic energy budget theory. We developed TKTD models with single and combined PMoAs to compare their goodness‐of‐fit and predicted population‐level sensitivity. We identified the PMoA reproduction efficiency as most probable in all species for Ni and Zn, but not for Cu, and found that combined‐PMoA models predicted higher population‐level sensitivity than single‐PMoA models, which was related to the predicted individual‐level sensitivity, rather than to mechanistic differences between models. Using point estimates of parameters, instead of sampling from the probability distributions of parameters, could also lead to differences in the predicted population‐level sensitivity. According to model predictions, apical chronic endpoints (cumulative reproduction, survival) are conservative for single‐metal population effects across metals and species. We conclude that the assumption of an identical PMoA for different species of Daphnia could be justified for Ni and Zn, but not for Cu. Single‐PMoA models are more appropriate than combined‐PMoA models from a model selection perspective, but propagation of the associated uncertainty should be considered. More accurate predictions of effects at low concentrations may nevertheless motivate the use of combined‐PMoA models. Environ Toxicol Chem 2024;43:338–358. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. [ABSTRACT FROM AUTHOR]

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Ecosystems are connected by flows of nutrients and organisms. Changes to connectivity and nutrient enrichment may destabilise ecosystem dynamics far from the nutrient source.We used gradostats to examine the effects of trophic connectivity (movement of consumers and producers) versus nutrient‐only connectivity on the dynamics of Daphnia pulex (consumers) and algae (resources) in two metaecosystem configurations (linear vs. dendritic).We found that Daphnia peak population size and instability (coefficient of variation; CV) increased as distance from the nutrient input increased, but these effects were lower in metaecosystems connected by all trophic levels compared with nutrient‐only connected systems and/or in dendritic compared with linear systems.We examined the effects of trophic connectivity (i.e. both trophic levels move rather than one or the other) using a generic model to qualitatively assess whether the expectations align with the ecosystem dynamics we observed.Analysis of our model shows that increased Daphnia population sizes and fluctuations in consumer‐resource dynamics are expected with nutrient connectivity, with this pattern being more pronounced in linear rather than dendritic systems.These results confirm that connectivity may propagate and even amplify instability over a metaecosystem to communities distant from the source disturbance, and suggest a direction for future experiments, that recreate conditions closer to those found in natural systems. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

Morphological variation is sometimes used as an indicator of environmental stress in animals. Here, we assessed how multiple morphological traits covaried in Daphnia pulex exposed to five common forms of environmental stress (high temperature, presence of predator cues, high salinity, low food abundance, and low Ca). We measured animal body length, body width, head width, eyespot diameter, and tail spine length along with mass in animals of five different ages (3, 6, 9, 12, and 15 d). There were strong allometric relationships among all morphological traits in reference animals and strong univariate effects of environmental stress on body mass and body length. We found that environmental stressors altered bivariate relationships between select pairwise combinations of morphological traits, with effects being dependent on animal age. Multivariate analyses further revealed high connectivity among body size–related traits but that eyespot diameter and tail spine length were less tightly associated with body size. Animals exposed to natural lake water with and without supplemental food also varied in morphology, with body size differences being suggestive of starvation and other unknown nutritional deficiencies. Yet our results demonstrate that the scaling of body morphological traits of Daphnia pulex is largely invariant with possible context-dependent plasticity in eye size and tail spine lengths. The strong coordination of traits indicates tight molecular coordination of body size during development despite strong and varied environmental stress. [ABSTRACT FROM AUTHOR]

Although the nutritional demands of animals are known to differ between immature and mature stages and between neonates from different clutches in various organisms, little evidence has been accumulated to examine this possibility in crustacean zooplankters. To this end, we examined the five digestive enzyme activities related to assimilations of carbohydrates, lipids, amino acids and phosphorus in the neonates, immature and mature individuals of panarctic Daphnia pulex. We found that all the enzyme activities were higher at lower food levels, indicating that the activities increased with decreasing supplies of the nutritional substances relative to the demands. As expected, these enzyme activities changed ontogenetically. Beta‐glucosidase activities increased with age, especially at a high food level, while the opposite pattern was found for lipase activities. A decreasing trend with age was also found in the activities of arginine and alanine amino‐peptidase and alkaline phosphatase at a low food level. These results suggest that juvenile individuals require more lipids, amino acids and phosphorus relative to the requirements of adult individuals, while the opposite pattern was true for carbohydrates. We also measured the enzyme activities of the neonates born from the 1st and 2nd clutches of the maternal individuals. The results showed that most digestive enzyme activities differed significantly between the neonates from the first and second clutches depending on the food levels. The present study confirmed that the nutritional demands differ not only between mature and immature individuals in Daphnia pulex and suggests that the mature individuals invest nutrients and energy reserves differently to neonates of the different clutches depending on their nutritional conditions. [ABSTRACT FROM AUTHOR]

Studies of closely related species with known ecological differences provide exceptional opportunities for understanding the genetic mechanisms of evolution. In this study, we compared population-genomics data between Daphnia pulex and Daphnia pulicaria , two reproductively compatible sister species experiencing ecological speciation, the first largely confined to intermittent ponds and the second to permanent lakes in the same geographic region. Daphnia pulicaria has lower genome-wide nucleotide diversity, a smaller effective population size, a higher incidence of private alleles, and a substantially more linkage disequilibrium than D. pulex. Positively selected genes in D. pulicaria are enriched in potentially aging-related categories such as cellular homeostasis, which may explain the extended life span in D. pulicaria. We also found that opsin-related genes, which may mediate photoperiodic responses, are under different selection pressures in these two species. Genes involved in mitochondrial functions, ribosomes, and responses to environmental stimuli are found to be under positive selection in both species. Additionally, we found that the two species have similar average evolutionary rates at the DNA-sequence level, although approximately 160 genes have significantly different rates in the two lineages. Our results provide insights into the physiological traits that differ within this regionally sympatric sister-species pair that occupies unique microhabitats. [ABSTRACT FROM AUTHOR]

The field of genomics has ushered in new methods for studying molecular-genetic variation in natural populations. However, most population-genomic studies still rely on small sample sizes (typically, <100 individuals) from single time points, leaving considerable uncertainties with respect to the behavior of relatively young (and rare) alleles and, owing to the large sampling variance of measures of variation, to the specific gene targets of unusually strong selection. Genomic sequences of ∼1,700 haplotypes distributed over a 10-year period from a natural population of the microcrustacean Daphnia pulex reveal evolutionary-genomic features at a refined scale, including previously hidden information on the behavior of rare alleles predicted by recent theory. Background selection, resulting from the recurrent introduction of deleterious alleles, appears to strongly influence the dynamics of neutral alleles, inducing indirect negative selection on rare variants and positive selection on common variants. Temporally fluctuating selection increases the persistence of nonsynonymous alleles with intermediate frequencies, while reducing standing levels of variation at linked silent sites. Combined with the results from an equally large metapopulation survey of the study species, classes of genes that are under strong positive selection can now be confidently identified in this key model organism. Most notable among rapidly evolving Daphnia genes are those associated with ribosomes, mitochondrial functions, sensory systems, and lifespan determination. [ABSTRACT FROM AUTHOR]

The taxonomic composition of the microbiota in the gut and epidermis of animals is known to vary among genetically and physiologically different host individuals within the same species. However, it is not clear whether the taxonomic composition diverges with increasing genetic distance of the host individuals. To unveil this uncertainty, we compared the host-associated microbiota among the genotypes within and between genetically distant lineages of parthenogenetic Daphnia cf. pulex across different physiological states, namely, well-fed, starved, and dead. Metagenomic analysis with 16S rRNA showed that, regardless of the host genotypes, diversity of the host-associated microbiota was high when the host individuals were fed food and gradually decreased when they were starved until they died. However, the difference in the host-associated microbiota, that is, β-diversity, was significant among the genotypes within and between the host lineages when they were fed. Although some bacteria in the microbiota, such as Limnohabitans, Rhodococcus, and Aeromicrobium, were found abundantly and commonly in all host genotypes; others, such as those of Holosoporacea, were found only in the genotypes of a specific lineage. Accordingly, the β-diversity tended to increase with increasing genetic distance of the host individuals. These results support an idea that the host-associated microbiota diverged with genetic divergence in the host species and that at least some bacteria are highly dependent on the genetically specific metabolites produced by the host individuals. [ABSTRACT FROM AUTHOR]

Obligate parthenogenesis (OP) is often thought to evolve by disruption of reductional meiosis and suppression of crossover recombination. In the crustacean Daphnia pulex , OP lineages, which have evolved from cyclical parthenogenetic (CP) ancestors, occasionally produce males that are capable of reductional meiosis. Here, by constructing high-density linkage maps, we find that these males show only slightly and nonsignificantly reduced recombination rates compared to CP males and females. Both meiosis disruption and recombination suppression are therefore sex-limited (or partly so), which speaks against the evolution of OP by disruption of a gene that is essential for meiosis or recombination in both sexes. The findings may be explained by female-limited action of genes that suppress recombination, but previously identified candidate genes are known to be expressed in both sexes. Alternatively, and equally consistent with the data, OP might have evolved through a reuse of the parthenogenesis pathways already present in CP and through their extension to all events of oogenesis. The causal mutations for the CP to OP transition may therefore include mutations in genes involved in oogenesis regulation and may not necessarily be restricted to genes of the "meiosis toolkit." More generally, our study emphasizes that there are many ways to achieve asexuality, and elucidating the possible mechanisms is key to ultimately identify the genes and traits involved. [ABSTRACT FROM AUTHOR]

1. The influence of cyanobacteria on interspecies interactions has received growing attention as a predictor of aquatic ecosystem function. It is well‐known that cyanobacteria greatly affect predator‐induced phenotypic plasticity in planktonic animals. However, the underlying molecular mechanism remains unclear. 2. Here, we used Daphnia pulex as a model organism for inducible defence, examining its morphological and life‐history responses to fish kairomone while feeding on 100% Chlorella pyrenoides, or while feeding on 70% Chlorella and either microcystin‐producing or ‐free Microcystis aeruginosa, separately. Transcriptome profiles of kairomone‐exposed D. pulex fed different types of food were detected using RNA‐seq. How the differentially expressed genes and pathways identified are responsible for the altered adaptive traits under Microcystis stress was discussed. 3. Daphnia pulex fed 100% Chlorella matured at a smaller body size, elongated tail spine length, and produced smaller offspring as adaptive morphological and reproductive responses to fish kairomone. These anti‐predation defences entailed a cost in decreased somatic growth rate. Microcystis stress caused defence trade‐offs in both morphology and reproduction: reduction of body size at maturity was stronger whereas tail spine elongation was unchanged or inhibited. Given that reproductive investment was reduced overall in Daphnia exposed to fish kairomones, there was a trade‐off for unchanged offspring size by reducing offspring number in Daphnia fed Microcystis. Defence‐induced costs to growth were increased by Microcystis exposure. 4. Transcriptome analysis revealed that neuronal signals including acetylcholine and glutamate signalling implicated in kairomone reception and/or transmission exhibited stronger responses under Microcystis stress. Cuticle development associated with the biosynthesis of wax esters and chitin, modification of lipo‐chitin saccharides, and the stability of collagen triple helix apparently participated in the morphological changes. The arachidonic acid pathway and biosynthesis of cholesterol and steroid hormones mediated altered reproductive performance. In addition, metabolic processes such as detoxification, food assimilation, lysosome dysfunction and apoptosis activation were involved in the survival and growth of Daphnia, which may contribute to the increased defence‐induced cost in growth when fed Microcystis. 5. Taken together, these findings advance our understanding of the molecular mechanisms underlying anti‐predation defensive responses by Daphnia to cyanobacterial stress. This work also provides a reference for further exploring the functional genes that mediate zooplankton–fish coevolution dynamics under eutrophic conditions. [ABSTRACT FROM AUTHOR]

Cyclical parthenogenesis, where females can engage in sexual or asexual reproduction depending on environmental conditions, represents a novel reproductive phenotype that emerged during eukaryotic evolution. The fact that environmental conditions can trigger cyclical parthenogens to engage in distinct reproductive modes strongly suggests that gene expression plays a key role in the origin of cyclical parthenogenesis. However, the genetic basis underlying cyclical parthenogenesis remains understudied. In this study, we characterize the female transcriptomic signature of sexual versus asexual reproduction in the cyclically parthenogenetic microcrustacean Daphnia pulex and Daphnia pulicaria. Our analyses of differentially expressed genes (DEGs), pathway enrichment, and gene ontology (GO) term enrichment clearly show that compared with sexual reproduction, the asexual reproductive stage is characterized by both the underregulation of meiosis and cell cycle genes and the upregulation of metabolic genes. The consensus set of DEGs that this study identifies within the meiotic, cell cycle, and metabolic pathways serves as candidate genes for future studies investigating how the two reproductive cycles in cyclical parthenogenesis are mediated at a molecular level. Furthermore, our analyses identify some cases of divergent expression among gene family members (e.g. doublesex and NOTCH2) associated with asexual or sexual reproductive stage, suggesting potential functional divergence among gene family members. [ABSTRACT FROM AUTHOR]

Limits of thermal tolerance in animal life is dependent on energy supply. Accordingly, the lowered ATP production capacity in ectotherms at high temperatures, which arises from a mismatch between oxygen supply and demand and the consequent switch from aerobic to anaerobic metabolism, affects the thermal resistance of these animals. The anaerobic ATP production capacity depends on the functional properties of the enzymes that reduce pyruvate. Thus, the present study focused on the role of the lactate dehydrogenase (LDH) of two daphnid species for anaerobic energy production at warm temperatures and the implications for their specific heat tolerances. Daphnia magna showed a higher thermal limit (indicated by immobilization time at 37°C) than Daphnia pulex, and in both species, this limit increased with rising acclimation temperature. In contrast to D. pulex, D. magna accumulated significant amounts of lactate at higher ambient temperatures. The intensity of anaerobic metabolism was also affected by acclimation temperature. Studying the functional enzyme properties revealed altered maximal reaction rates and substrate inhibitions of the LDH suites of the two daphnid species. D. magna LDH showed a significantly lower substrate inhibition than D. pulex LDH. The LDH isoform composition and the temperature‐induced changes differed between both species. The detected qualitative modulations of the LDH suites may have resulted from differential isoform expression and different maturation processes. The species‐specific LDH characteristics imply a higher anaerobic energy production at warm temperatures in D. magna, which likely contributes to the higher heat tolerance of this species. Research highlights: Heat tolerance in Daphnia is related tolactate production.Acclimation temperature affects LDH.LDH substrate inhibition prevents high lactate formation.Anaerobic LDH capacity is higher in Daphniamagna than in Daphniapulex.Differentia lLDH isoform expression adds to heat tolerance. [ABSTRACT FROM AUTHOR]

In this work, we developed and used research methods that will make it possible to quickly assess the toxic activity of, among others, metal nanoparticles. In this work, we assessed the toxicity of PbO nanoparticles obtained from biological synthesis using Farfarae folium. The obtained PbO nanoparticles were characterized with the use of such methods as UV–VIS, XRD, SEM with EDS, AFM and FTIR. The techniques applied in the work confirmed the presence of 30–40 nm PbO nanoparticles. The toxic properties of biologically synthesized nanoparticles were assessed with the use of plants and invertebrates. The phytotoxicity studies were conducted on the following plant seeds: radish, garden cress, bean, rape, chia, lupine, corn, and beetroot. The IC50 coefficient was calculated on the basis of the logarithmic relationship Aw = a*ln(C) + b. The studies also used the following invertebrates: Daphnia pulex, Tubifex tubifex, Chaoborus sp. (larva), Chironomus aprilinus, Artemia sp. The EC50 coefficient was calculated after the incubation period. For PbO nanoparticles, we managed to identify the IC50 coefficient for selected plants. It was, respectively, 0.150 mg/ml for lupine; 0.274 mg/ml for bean; and 0.655 mg/ml for radish. As regards the studies on invertebrates, it was observed that the most sensitive organisms were Daphnia pulex (EC50 = 0.32 mg/ml) and Tubifex tubifex (EC50 = 0.38 mg/ml), and the most resistant one was Chironomus aprilinus (EC50 = 0.83 mg/ml). [ABSTRACT FROM AUTHOR]

Anthropogenic noise has the potential to alter community dynamics by modifying the strength of nested ecological interactions such as predation. Direct effects of noise on per capita predation rates have received much attention but the context in which predation occurs is often oversimplified. For instance, many animals interact with conspecifics while foraging and these nontrophic interactions can positively or negatively influence per capita predation rates. These effects are often referred to as multiple‐predator effects (MPEs). The extent to which noise can modulate MPEs and thereby indirectly alter per capita predation remains unknown. To address this question, we derived the relationship between per capita predation rate and prey density, namely the functional response (FR), of single and pairs of the invasive topmouth gudgeon Pseudorasbora parva when feeding on water fleas under two noise conditions: control ambient noise estimated at 95 dB re 1 μPa and ambient noise supplemented with motorboat sounds whose relative importance over ambient noise ranged from 4.81 to 27 dB. In addition, we used video recordings to track fish movements. To detect MPEs, we compared the observed group‐level FRs to predicted group‐level FRs inferred from the individual FRs and based on additive effects only. Regardless of the number of fish and the noise condition, the FR was always of type II, showing predation rate in a decelerating rise to an upper asymptote. Compared to the noiseless condition, the predation rate of single fish exposed to noise did not differ at high prey densities but was significantly lower at low prey densities, resulting in an FR with the same asymptote but a less steep initial slope. Noise also reduced fish mobility, which might explain the decrease in predation rate at low prey densities. Conspecific presence suppressed the individual response to noise, the FRs of two fish (observed group‐level FRs) being perfectly similar between the two noise conditions. Although observed and predicted group‐level FRs did not differ significantly, observed group‐level FRs tended to fall in the low range of predicted group‐level FRs, suggesting antagonism and a negative effect of nontrophic interactions on individual foraging performance. Interestingly, the difference between predicted and observed group‐level FRs was not greater with noise, which means that noise did not strengthen MPEs. Our results show that when considering the social context of foraging, here through the presence of a conspecific, anthropogenic noise does not compromise foraging in the invasive P. parva. [ABSTRACT FROM AUTHOR]

Zooplankton body size shows a strong association with temperature, competition, and predation. Global warming affects all three drivers of body size and is thus expected to lead to substantial changes in zooplankton community composition and body size distributions. To disentangle the isolated and joint effect of temperature, competition, and fish predation on species biomass and community composition in zooplankton, we monitored population biomasses of three Daphniidae species that differ in body size (Daphnia magna, Daphnia pulex, and Ceriodaphnia reticulata) for 20 days, manipulating competition (monoculture, pairwise trials, and three‐species communities), temperature (20°C, 24°C, and 28°C) and presence or absence of fish predation. In the absence of predation, D. magna dominated in all competition experiments, even at high temperatures. D. magna went extinct, however, in the predation treatments at 24°C and 28°C. D. pulex outcompeted C. reticulata and was negatively affected by predation and high temperature. C. reticulata did not reduce biomass at high temperatures and was negatively affected by all competition trials, but was positively affected by predation. Our results indicate that the two larger‐bodied species are more negatively affected by the combination of temperature and predation than the smallest species. While higher temperatures reduced the biomass of the larger‐bodied species, it did not fundamentally change their ability to dominate over the smallest species in competition. The combined effect of warming and predation changed community composition more fundamentally, resulting in the dominance of small‐bodied species. This can have important ecosystem‐wide impacts, such as the transition to turbid, algae‐dominated systems. [ABSTRACT FROM AUTHOR]

Background: The genus Daphnia switches its reproductive mode from subitaneous egg production to resting egg production in response to environmental stimuli. Although this life history trait is essential for surviving unsuitable environments, the molecular mechanism of resting egg production is little understood. In this study, we examined genes related to induction of resting egg production using two genotypes of panarctic Daphnia pulex, the JPN1 and JPN2 lineages, which differ genetically in the frequency of resting egg production. We reared these genotypes under high and low food levels. At the high food level, individuals of both genotypes continually produced subitaneous eggs, whereas at the low food level, only the JPN2 genotype produced resting eggs. Then, we performed RNA-seq analysis on specimens of three instars, including before and after egg production. Results: These results showed that expressed genes differed significantly between individuals grown under high and low food levels and among individuals of different instars and genotypes. Among these differentially expressed genes (DEGs), we found 16 that changed expression level before resting egg production. Some of these genes showed high-level expression only before resting egg production and one gene was an ortholog of bubblegum (bgm), which is reportedly up-regulated before diapause in bumblebees. According to gene ontology (GO) enrichment analysis, one GO term annotated as long-chain fatty acid biosynthetic process was enriched among these 16 genes. In addition, GO terms related to glycometabolism were enriched among down-regulated genes of individuals holding resting eggs, compared to those before resting egg production. Conclusions: We found candidate genes highly expressed only before resting egg production. Although functions of candidate genes found in this study have not been reported previously in Daphnia, catabolism of long-chain fatty acids and metabolism of glycerates are related to diapause in other organisms. Thus, it is highly probable that candidate genes identified in this study are related to the molecular mechanism regulating resting egg production in Daphnia. [ABSTRACT FROM AUTHOR]

Changes in day-length entrain the endogenous clock of organisms leading to complex responses to photoperiod. In long-lived organisms experiencing several seasons this response of the clock to photoperiod is phenotypically plastic. However, short-lived organisms often experience a single season without pronounced changes in day-length. For those, a plastic response of the clock to different seasons would not necessarily be adaptive. In aquatic ecosystems, zooplankton species like Daphnia live only for some weeks, i.e. one week up to ca. two months. However, they often show a succession of clones that are seasonally adapted to environmental changes. Here, we found that 16 Daphnia clones per each of three seasons (= 48 clones) from the same pond and year differed in clock gene expression with a homogenous gene expression pattern in ephippia-hatched spring clones and a bimodal expression pattern in summer and autumn populations indicating an ongoing adaptation process. We clearly demonstrate that spring clones were adapted to a short, and summer clones to a long photoperiod. Furthermore, we found that gene expression of the melatonin-synthesis enzyme AANAT was always lowest in summer clones. In the Anthropocene, Daphnia's clock might be disturbed by light-pollution and global warming. Since Daphnia is a key-organism in trophic carbon transfer, a disruption of its clock rhythm would be devastating for the stability of freshwater ecosystems. Our results are an important step in understanding the adaptation of Daphnia's clock to environmental changes. [ABSTRACT FROM AUTHOR]

Isothiazolinones are a family of broad‐spectrum biocides widely used in industry and consumer products. Chloro‐ and methyl‐isothiazolinones (CMIT and MIT) are documented as strong irritants, yet they are still used in a wide variety of applications, including cosmetics, cleansers, hygienic products, and various industrial applications. The subsequent substantial release of these molecules from urban sources into freshwater environments, and their potential impacts on aquatic species, have nevertheless received little attention so far, with few studies reporting on the toxicity of either CMIT or MIT to nontarget organisms. The present study addresses this current knowledge gap by evaluating the acute toxicity to Daphnia pulex (Cladocera) of CMIT/MIT (3:1) and MIT, the two formulations most commonly used by manufacturers. In addition, genetic diversity is known to be a major component of variability in phenotypic responses, although it is largely overlooked in typical toxicity tests. Thus the potential range of responses inherent to genetic diversity is rarely considered. Therefore, to account for intraspecific variations in sensitivity, our design involved eight clonal lines of D. pulex stemming from distinct natural populations or commercial strains. Clones exhibited strong variation in their responses, with median lethal concentration (LC50) values ranging from 0.10 to 1.84 mg/L for the mixture CMIT/MIT, and from 0.68 to 2.84 mg/L for MIT alone. These intraspecific ranges of LC50 values challenge the use of single clones of daphnids in standard ecotoxicological tests and the predictions based on their results. The present study brings new evidence that assessing ecological risk of chemicals while ignoring genotype diversity is neither ecologically relevant, nor a representative evaluation of the diversity of potential adverse outcomes. Environ Toxicol Chem 2023;42:805–814. © 2023 SETAC [ABSTRACT FROM AUTHOR]

Our previous studies showed that the change in the plankton response to light could be an indicator of environmental pollution. This study experimentally reveals that the response of Daphnia magna Straus and Daphnia pulex plankton ensembles to photostimulation depends on the intensity of the attracting light. This makes it difficult to identify the occurrence and change of pollutant concentration. The large variability in the magnitude of the behavioral response is caused by the nonlinear response of plankton ensembles to the intensity of the attractor stimulus. As the intensity of the photostimulation increases, the variability of the phototropic response passes through increase, decrease, and relative stabilization phases. The paper proposes a modification of the photostimulation method—paired photostimulation involving successive exposure to two photostimuli of increasing intensity. The first stimulus stabilizes the behavioral response, while the increase in response to the second stimulus makes it possible to more accurately assess the responsiveness of the plankton ensemble. The paper studies the sensitivity of the method of paired stimulation of the behavioral response of different types of freshwater plankton ensembles: Daphnia magna Straus, Daphnia pulex to the effects of pollutants (potassium bichromate, microplastic). The study demonstrates good reliability and increased sensitivity of this method of detecting changes in environmental toxicity compared to single photostimulation or traditional bioindication through the survival rate of test organisms. [ABSTRACT FROM AUTHOR]

Speciation genomic studies have revealed that genomes of diverging lineages are shaped jointly by the actions of gene flow and selection. These evolutionary forces acting in concert with processes such as recombination and genome features such as gene density shape a mosaic landscape of divergence. We investigated the roles of recombination and gene density in shaping the patterns of differentiation and divergence between the cyclically parthenogenetic ecological sister‐taxa, Daphnia pulicaria and Daphnia pulex. First, we assembled a phased chromosome‐scale genome assembly using trio‐binning for D. pulicaria and constructed a genetic map using an F2‐intercross panel to understand sex‐specific recombination rate heterogeneity. Finally, we used a ddRADseq data set with broad geographic sampling of D. pulicaria, D. pulex, and their hybrids to understand the patterns of genome‐scale divergence and demographic parameters. Our study provides the first sex‐specific estimates of recombination rates for a cyclical parthenogen, and unlike other eukaryotic species, we observed male‐biased heterochiasmy in D. pulicaria, which may be related to this somewhat unique breeding mode. Additionally, regions of high gene density and recombination are generally more divergent than regions of suppressed recombination. Outlier analysis indicated that divergent genomic regions are probably driven by selection on D. pulicaria, the derived lineage colonizing a novel lake habitat. Together, our study supports a scenario of selection acting on genes related to local adaptation shaping genome‐wide patterns of differentiation despite high local recombination rates in this species complex. Finally, we discuss the limitations of our data in light of demographic uncertainty. [ABSTRACT FROM AUTHOR]

The ability to directly measure and monitor poor nutrition in individual animals and ecological communities is hampered by methodological limitations. In this study, we use nutrigenomics to identify nutritional biomarkers in a freshwater zooplankter, Daphnia pulex, a ubiquitous primary consumer in lakes and a sentinel of environmental change. We grew animals in six ecologically relevant nutritional treatments: nutrient replete, low carbon (food), low phosphorus, low nitrogen, low calcium and high Cyanobacteria. We extracted RNA for transcriptome sequencing to identify genes that were nutrient responsive and capable of predicting nutritional status with a high degree of accuracy. We selected a list of 125 candidate genes, which were subsequently pruned to 13 predictive potential biomarkers. Using a nearest‐neighbour classification algorithm, we demonstrate that these potential biomarkers are capable of classifying our samples into the correct nutritional group with 100% accuracy. The functional annotation of the selected biomarkers revealed some specific nutritional pathways and supported our hypothesis that animal responses to poor nutrition are nutrient specific and not simply different presentations of slow growth or energy limitation. This is a key step in uncovering the causes and consequences of nutritional limitation in animal consumers and their responses to small‐ and large‐scale changes in biogeochemical cycles. [ABSTRACT FROM AUTHOR]

Daphnia pulex is a natural food widely used in aquaculture, and S6K plays an important role in protein translation and ageing. To explore the function of S6K in the ageing process of D. pulex , we cloned and analysed the full-length cDNA of S6K. With the growth of D. pulex , the expression of S6K mRNA first increased, then peaked at 10 days, and then decreased. Western blot analysis showed that the S6K protein expression increased from 1 to 5 days of age, but did not change significantly from 5 to 25 days of age. RNAi knockdown of S6K, reduced the expression of S6K mRNA and S6K protein, and it causes changes in other genes associated with ageing in both sexes. S6K may be involved in reproductive transformation, affecting reproductive capacity, thus significantly affecting the ageing of D. pulex. miRNA with potential regulation of the S6K gene were screened by the bioinformatics method and validated using a dual luciferase reporter gene system. This study deepens our understanding of the mechanisms of ageing in D. pulex and expands our understanding of the role of S6K in the ageing process. [ABSTRACT FROM AUTHOR]

Abstract Water fleas of the genus Daphnia have been a model system for hundreds of years and is among the best studied ecological model organisms to date. Daphnia are planktonic crustaceans with a cyclic parthenogenetic life-cycle. They have a nearly worldwide distribution, inhabiting standing fresh- and brackish water bodies, from small temporary pools to large lakes. Their predominantly asexual reproduction allows for the study of phenotypes excluding genetic variation, enabling us to separate genetic from non-genetic effects. Daphnia are often used in studies related to ecotoxicology, predator-induced defence, host–parasite interactions, phenotypic plasticity and, increasingly, in evolutionary genomics. The most commonly studied species are Daphnia magna and D. pulex, for which a rapidly increasing number of genetic and genomic tools are available. Here, I review current research topics, where the Daphnia model system plays a critical role.

Rising salinity from road deicing salts threatens the survival and reproduction of freshwater organisms. We conducted two experiments to address how Daphnia pulex survival and reproduction were affected by road salt concentration (control, 120, 640 and 1200 mg Cl−/L) crossed with three concentrations of water hardness (20, 97, 185 mg CaCO3 /L). D. pulex survival was poor in our hard water treatment in both experiments (185 mg CaCO3 /L), potentially indicating a low tolerance to hard water for the strain used in our experiments. With the remaining two hardness treatments (20 and 97 mg CaCO3 /L), we found no evidence of an interactive effect between salt concentration and water hardness on D. pulex survival. In our population-level experiment, D. pulex survival was reduced by > 60% at 120 mg Cl−/L compared to the control. In the individual experiment, survival was similar between the control and 120 mg Cl−/L, but ≤ 40% of individuals survived in 640 and 1200 mg Cl−/L. For the surviving individuals across all treatments, the number of offspring produced per individual declined with increasing Cl− concentration and in hard water. Our results indicate that current Cl− thresholds may not protect some zooplankton and reduced food availability per capita may enhance the negative impacts of road salt. [ABSTRACT FROM AUTHOR]

Kairomone‐induced anti‐predation defences in zooplankton mediate trophic interactions in aquatic ecosystems. Changes in food quality greatly affect induced defences because of the potential influence on resource allocation. Understanding how zooplankton respond to predation risk under poor nutrition conditions is critical for predicting ecosystem function.The present study focused on the fish kairomone‐induced defences in Daphnia pulex fed unicellular non‐toxic Microcystis aeruginosa. Transcriptions of key genes were detected to interpret changes in morphological and life history traits. Additionally, the degree of responsiveness of traits to increasing proportions of M. aeruginosa in the diet was compared at different endpoints with varied exposure duration.Daphnia pulex decreased body size but showed an elongated tail spine as morphological adaptations to the presence of fish kairomones, supported by enhanced transcriptions of chitin deacetylase and cuticle protein genes. However, with suppressed expression of the related genes, tail spine length was linearly reduced by increasing proportions of M. aeruginosa in the diet, accompanied by decreased somatic growth and reproduction. Presence of fish kairomones interactively affected the inhibitive effects of increasing proportions of M. aeruginosa in the diet on Daphnia growth, reproduction, and tail spine defence but not body size. In addition, the magnitude of effects of M. aeruginosa on Daphnia growth and morphological responses under fish kairomone exposure was increased with the increasing length of exposure time from 3–7 days (reaching maturity) to 14 days (by the end of tests), indicating an aggravated inhibitive effect of M. aeruginosa on induced defences over time.The present study provides references for evaluating zooplankton‐fish interactions under poor nutrition conditions. Since a cyanobacterial bloom may last from a few weeks to several months, experiments involving the whole life span of zooplankton are thought to fully reveal the actual impact of cyanobacteria on interspecific interactions related to zooplankton. In addition, unifying the exposure time to cyanobacteria for comparative studies is recommended in future research. [ABSTRACT FROM AUTHOR]

The recovery of sludge produced in the wastewater treatment process in WWTPs is often limited by the high content of toxic forms of contaminants of both an inorganic and organic nature. One of the options for the effective treatment of the world's ever-increasing quantities of sewage sludge is the pyrolysis process. Thermochemical conversion of sewage sludge is emerging as a promising method for treating these heterogeneous and highly complex wastes with increasing research work. Pyrolysis-treated sewage sludge (PM) prepared at 603–615 °C was characterized by pH, EC, and CHN-S analysis; total and bioavailable concentrations of P and heavy metals (Cd, Cu, Fe and Zn); fractionation of bound forms of P and heavy metals in the material and determination of the presence of polycyclic aromatic hydrocarbons (PAHs). The studied material was subjected to ecotoxicological tests (Daphnia pulex L.) and cultivation tests (Lactuca sativa L.). Elemental analysis revealed the concentrations of heavy metals in PM: Fe (137,600 mg/kg), Zn (2602 mg/kg), Cu (582 mg/kg), Cr (107 mg/kg), Pb (87 mg/kg), Ni (67 mg/kg), As (<1 mg/kg), Hg (<2 mg/kg) and Cd (<1 mg/kg). The highest values of extractability of the investigated heavy metals from PM were found in the cases of Zn (HCl) and Fe (Mehlich 3), both values not exceeding 500 mg/kg. BCR sequential extraction showed the major concentrations of Cu and Fe were predominantly bound in the residual fraction (F4) and Zn in the reducible fraction (F2) of PM. The results of heavy metal bioavailability suggest that the addition of PM does not negatively affect the growth of lettuce biomass and the metal contents of plant tissues. Based on the results obtained, the pyrolysis material prepared from municipal sewage sludge seems to be a promising and innovative soil additive and a potential alternative to conventional inorganic fertilizers. [ABSTRACT FROM AUTHOR]

Climate change is increasing water temperature and intensifying the incidence of cyanobacterial blooms worldwide. However, the combined effects of increased temperature and microcystin concentrations as co-stressors on survival and ecological processes in freshwater species are unclear. Here, using purified MC-LR and crude extract of toxigenic Microcystis aeruginosa, we tested the individual and combined effects of three water temperatures (15, 20, 25 °C) and a range of environmentally relevant concentrations of dissolved microcystin and crude extract (0.01–10 µg·L−1) on survival, growth inhibition, grazing and predation rates in three freshwater species: phytoplankton (Scenedesmus quadricauda), zooplankton (Daphnia pulex), and an invertebrate predator (Ischnura elegans). Purified MC-LR exerted a higher growth inhibitory effect on S. quadricauda compared to crude extract with the same concentration of MC-LR, while neither treatment affected its chlorophyll-a content or survival of D. pulex. Crude extract reduced grazing and survival of D. pulex and I. elegans, respectively. The combined effect of higher temperature and crude extract reduced I. elegans survival by 50%. Increased temperature reduced prey handing time in I. elegans by 49%, suggesting a higher predation rate. However, warming together with higher concentrations of crude extract jointly increased zooplankton grazing and reduced damselfly predation. Taken together, these results suggest crude extract, and not necessarily microcystin, can affect survival and productivity in freshwater species, although these effects may vary unevenly across trophic levels. Our findings highlight the importance of complex ecological mechanisms by which warming can exacerbate toxic effects of cyanobacterial bloom extracts on survival and functions among species in eutrophic freshwaters. [ABSTRACT FROM AUTHOR]

Methyl farnesoate (MF), a juvenile hormone, can influence phenotypic traits and stimulates male production in daphnids. MF is produced endogenously in response to stressful conditions, but it is not known whether this hormone can also be released into the environment to mediate stress signaling. In the present study, for the first time, a reliable solid-phase microextraction–gas chromatography–mass spectrometry (SPME-GC-MS) method was developed and validated for the ultra-trace analysis of MF released in growth medium by Daphnia pulex maintained in presence of crowding w/o MK801, a putative upstream inhibitor of MF endogenous production. Two different clonal lineages, I and S clones, which differ in the sensitivity to the stimuli leading to male production, were also compared. A detection limit of 1.3 ng/L was achieved, along with good precision and trueness, thus enabling the quantitation of MF at ultra-trace level. The achieved results demonstrated the release of MF by both clones at the 20 ng/L level in control conditions, whereas a significant decrease in the presence of crowding was assessed. As expected, a further reduction was obtained in the presence of MK801. These findings strengthen the link between environmental stimuli and the MF signaling pathway. Daphnia pulex, by releasing the juvenile hormone MF in the medium, could regulate population dynamics by means of an autoregulatory feedback loop that controls the intra- and extra-individual-level release of MF produced by endogenous biosynthesis. [ABSTRACT FROM AUTHOR]

Abstract It has been hypothesized that the effects of pollutants on phenotypes can be passed to subsequent generations through epigenetic inheritance, affecting populations long after the removal of a pollutant. But there is still little evidence that pollutants can induce persistent epigenetic effects in animals. Here, we show that low doses of commonly used pollutants induce genome‐wide differences in cytosine methylation in the freshwater crustacean Daphnia pulex. Uniclonal populations were either continually exposed to pollutants or switched to clean water, and methylation was compared to control populations that did not experience pollutant exposure. Although some direct changes to methylation were only present in the continually exposed populations, others were present in both the continually exposed and switched to clean water treatments, suggesting that these modifications had persisted for 7 months (>15 generations). We also identified modifications that were only present in the populations that had switched to clean water, indicating a long‐term legacy of pollutant exposure distinct from the persistent effects. Pollutant‐induced differential methylation tended to occur at sites that were highly methylated in controls. Modifications that were observed in both continually and switched treatments were highly methylated in controls and showed reduced methylation in the treatments. On the other hand, modifications found just in the switched treatment tended to have lower levels of methylation in the controls and showed increase methylation in the switched treatment. In a second experiment, we confirmed that sublethal doses of the same pollutants generate effects on life histories for at least three generations following the removal of the pollutant. Our results demonstrate that even low doses of pollutants can induce transgenerational epigenetic effects that are stably transmitted over many generations. Persistent effects are likely to influence phenotypic development, which could contribute to the rapid adaptation, or extinction, of populations confronted by anthropogenic stressors.

The discharge of metal nanoparticles into the water inevitably poses a threat to aquatic organisms and the balance of the aquatic ecosystem. Photoperiod is one of the most important ecological factors for the development of cladocerans. In addition, different light conditions can also affect the toxicity of metal nanoparticles. In this study, we studied the effects of four photoperiods (8L/16D, 10L/14D, 14L/10D, and 16L/8D) combined with three concentrations of ZnO NPs (0 mg L−1, 0.05 mg L−1, and 0.10 mg L−1) on the growth and reproduction of Daphnia pulex. With the increase of photoperiod, the maternal body size and growth rate increased first and then decreased; the first time to reproduction was advanced, and broods and the total offspring also increased. Under the influence of ZnO NPs, growth rate and reproductive capacity were inhibited. The photoperiod 8L/16D and 16L/8D interacted with ZnO NPs on the growth of D. pulex , which significantly decreased the growth rate. Besides, the interaction between photoperiod 8L/16D and ZnO NPs decreased the reproduction ability of D. pulex. These results suggest that the effects of zinc oxide nanoparticles on the growth and reproduction of D. pulex is photoperiod dependent, which is useful for assessing the risk of pollutants to cladoceras under different light conditions. [Display omitted] • Moderate lighting time is suitable for the growth of D. pulex. • Long lighting time is conducive to reproductive ability of D. pulex. • ZnO NPs inhibit the growth and reproduction of D. pulex. • Long or short lighting time intensify inhibitory effect of ZnO NPs on growth. • Long lighting time reduces the inhibitory effect of ZnO NPs on reproduction. [ABSTRACT FROM AUTHOR]