Toxicity of benzethonium chloride and polyhexamethylene guanidine hydrochloride mixtures on Daphnia carinata: synergistic and antagonistic effects at specific ratios.

Throughout the coronavirus (COVID-19) pandemic, the sanitizing products benzethonium chloride (BEC) and polyhexamethylene guanidine hydrochloride (PHMG-H) were widely used; however, few studies have investigated their combined toxicity to organisms. In the present study, acute toxicity and genotoxicity of BEC, PHMG-H, and the combination of the two were investigated as endpoints using Daphnia carinata as the model organism. For individual reagents, PHMG-H was found to be more toxic than BEC in terms of both mortality and genotoxicity. DNA damage and survival rate were used as toxicity endpoints. The interaction was evaluated with the concentration addition (CA) model via toxic unit (TU) approach and additive index (AI) method in mixtures at different ratios in TU. Only the binary mixture BEC + PHMG-H at the ratio 1:9 in TU was regarded as synergistic, while all others indicated increased antagonistic effects as the proportion of BEC increased over the PHMG-H concentration. The findings here benefit understanding surrounding precisely how BEC and PHMG-H interact at different mixing ratios, and can assist with the evaluation of risk assessments for binary mixtures in aquatic ecosystems.

Cascading effects caused by fenoxycarb in freshwater systems dominated by Daphnia carinata and Dolerocypris sinensis.

To evaluate the aquatic hazards of the insect juvenile hormone analogue fenoxycarb, a single application (0, 48.8, 156.3, 500, 1600, and 5120 µg/L) of it was done in indoor freshwater systems dominated by Daphnia carinata (daphnid) and Dolerocypris sinensis (ostracoda). The responses of zooplankton (counted by abundance and the activity and immuno-reactive content of free N-Acetyl-ß-D-glucosaminidase (NAGase)), phytoplankton (counted by chlorophyll and phycocyanin), planktonic bacteria and fungi, and some water quality parameters were investigated in a period of 35 d. Results of the study showed that the ostracoda was more sensitive than daphnid, with time-weighted average (TWA)-based no observed effect concentrations (NOECs) to be 8.45 and 12.66 µg/L in systems without humic acid addition (HA-) and to be 6.37 and 9.54 µg/L in systems with humic acid addition (HA+). The duration of treatment-related effects in the ostracoda population was longer than the daphnid population (21 vs. 14 days). Besides, the data analysis indicated that the toxicity of fenoxycarb was significantly enhanced in the HA+ systems. Owing to the reduced grazing pressure, the concentrations of chlorophyll and phycocyanin increased in the two highest treatments. The increase in photosynthesis along with a reduced animal excretion led to an increase in pH and a decrease in nutrient contents. These changes seemed to have an effect on the microbial communities. For example, the abundances of some opportunistic pathogens of aquatic animals (e.g. Aeromonas and Cladosporium) and organic-pollutant-degrading microorganisms (e.g. Ancylobacter and Azospirillum) increased significantly in microbial communities, but the abundances of Pedobacter, Candidatus Planktoluna, and Rhodobacter (photosynthetic bacteria) markedly decreased. This study provides useful information to understand the ecotoxicological impacts of fenoxycarb at the population and community levels while integrating the effects of HA on toxicity.

Acute toxicity of the insecticide methyl parathion and its hydrolytic product p-nitrophenol to the native Australian cladoceran Daphnia carinata.

The toxicity of an organophosphorus (OP) insecticide, methyl parathion (MP), and its hydrolysis product, p-nitrophenol (PNP), to the native Australian cladoceran species, Daphnia carinata, was assessed. Both MP and PNP were stable in cladoceran water during the test period. D. carinata was sensitive to both MP and PNP; however, the parent compound was more toxic than its metabolite. This is the first study that demonstrated the acute toxicity of MP and PNP towards an Australian daphnid species. The present investigation emphasizes the need for including the native taxa as non-target test organisms while evaluating the toxicity of environmental pollutants.

The effect of food, light intensity and tank volume on resting eggs production in Daphnia carinata.

Sexual reproduction is an important stage of cyclical parthenogenesis Daphnia, these cyclic parthenogens need to time their shift from asexual to sexual reproduction correctly, based on environmental cues. We investigated the effect of food quality and quantity, light intensity, and culture tank volume on the resting eggs production of a tropical Daphnia carinata clone of Vietnam. The results showed that sexual ephippia production of the D. carinata was strongly induced when fed with green algae Scenedesmus sp., under moderate light intensity condition. The experiments also showed that food limitation is not an obligate requirement of shifting from parthenogenetic reproduction to sexual ephippia reproduction in D. carinata. Beside of that, culture volume seemed to be an important determinant of successfully forming of embryos inside ephippium. These findings here were from cultures in contaminant free condition, thus they are important basics for further studies of additional effects of environment toxicants on D. carinata species of Vietnam.

Genotoxicity assessment of acute exposure of 2, 4-dinitroanisole, its metabolites and 2, 4, 6-trinitrotoluene to Daphnia carinata.

The insensitive munition ingredient, 2, 4-dinitroanisole has emerged as an alternative ingredient to 2, 4, 6-trinitro toluene in melt pourable high explosive formulations mainly due to its improved insensitiveness properties. As a result, production of 2, 4-dinitroanisole has increased and as a consequence 2, 4-dinitroanisole has emerged as a potential ingredient to enter the environment and possibly persist in water and soil ecosystems. The present study showed that 2, 4-dinitroanisole, its metabolites (2-amino 4-nitroanisole and 2,4-dinitroanisole) and 2, 4, 6-trinitro toluene were found to induce DNA damages in a freshwater crustacean Daphnia carinata exposed for 48 h and which was investigated by the alkaline single-cell gel electrophoresis (comet assay) method. The value of LC50-48 h of 2, 4-dinitroanisole was determined as 14.87 ± 1.70 (mg L-1) and its metabolites exhibited the similar toxic range although the toxicity of 2, 4, 6-trinitro toluene was seven-fold more toxic (2.32 ± 0.29 mg L-1) than 2, 4-dinitroanisole and its metabolites. Exposure to sub-acute toxicity concentration ranges of 2, 4-dinitroanisole and its metabolites and 2, 4, 6-trinitro toluene showed significant (P < 0.01) DNA damage. The higher concentration of each test chemical exhibited higher tail DNA per cent and increased olive tail moment. The results from this study can be used to identify genotoxic biomarkers for the risk assessment of insensitive munitions exposure in aquatic invertebrates.

Exposure to perfluorooctanesulfonate (PFOS) but not perflurorooctanoic acid (PFOA) at ppb concentration induces chronic toxicity in Daphnia carinata.

Widespread environmental contamination of per- and polyfluoroalkyl substances (PFAS) is well established. Nevertheless, few studies have reported on the aquatic toxicity of PFAS, especially in indicator species such as Daphnia. In this study, the toxicity of two major PFAS, namely perfluorooctanoic acid (PFOA) and perfluorooctanesulfonate (PFOS), was investigated on water flea (Daphnia carinata) using a battery of comprehensive toxicity tests, including a 48 h acute and a 21-day chronic assays. The survival, growth, and reproduction of D. carinata were monitored over a 21-day life cycle. PFOS exhibited higher toxicity than PFOA. The 48 h LC50 values (confidence interval) based on acute toxicity for PFOA and PFOS were 78.2 (54.9-105) mg L-1 and 8.8 (6.4-11.6) mg L-1, respectively. Chronic exposure to PFOS for 21 days displayed mortality and reproductive defects in D. carinata at a concentration as low as 0.001 mg L-1. Genotoxicity assessment using comet assay revealed that exposure for 96 h to PFOS at 1 and 10.0 mg L-1 significantly damaged the organism's genetic makeup. The results of this study have great implications for risk assessment of PFOS and PFOA in aquatic ecosystems, given the potential of PFOS to pose a risk to Daphnia even at lower concentrations (1 µg L-1).

Anywhere but here: local conditions motivate dispersal in Daphnia.

Dispersal is fundamental to population dynamics. However, it is increasingly apparent that, despite most models treating dispersal as a constant, many organisms make dispersal decisions based upon information gathered from the environment. Ideally, organisms would make fully informed decisions, with knowledge of both intra-patch conditions (conditions in their current location) and extra-patch conditions (conditions in alternative locations). Acquiring information is energetically costly, however, and extra-patch information will typically be costlier to obtain than intra-patch information. As a consequence, theory suggests that organisms will often make partially informed dispersal decisions, utilising intra-patch information only. We test this proposition in an experimental two-patch system using populations of the aquatic crustacean, Daphnia carinata. We manipulated conditions (food availability) in the population's home patch, and in its alternative patch. We found that D. carinata made use of intra-patch information (resource availability in the home patch induced a 10-fold increase in dispersal probability) but either ignored or were incapable of using of extra-patch information (resource availability in the alternative patch did not affect dispersal probability). We also observed a small apparent increase in dispersal in replicates with higher population densities, but this effect was smaller than the effect of resource constraint, and not found to be significant. Our work highlights the considerable influence that information can have on dispersal probability, but also that dispersal decisions will often be made in only a partially informed manner. The magnitude of the response we observed also adds to the growing chorus that condition-dependence may be a significant driver of variation in dispersal.

In vivo Visualization of the Process of Hg2+ Bioaccumulation in Water Flea Daphnia carinata by a Novel Aggregation-Induced Emission Fluorogen.

This paper employs a specially designed aggregation-induced emission fluorogen (AIEgen) to in vivo visualize the process of Hg2+ bioaccumulation in a common species of freshwater zooplankton Daphnia carinata (D. carinata) by two methods, direct Hg2+ absorption and ingestion of Hg2+ contaminated food. We analysed the relevance between photoluminescence (PL, I595 /I480 ) ratios and Hg2+ (CHg2+ ) and developed the master curve for Hg2+ determination based on measuring the PL intensity of the solution. Meanwhile, fluorescent image analysis showed that the major recipient organs of Hg2+ in D. carinata were the compound eyes and carapace, followed by the intestine and shell gland, but not the brain or heart. The response of D. carinata to Hg2+ via uptake from surrounding water differed distinctly from that through food intake of the algae (Euglena gracilis) contaminated by mercury. When Hg2+ was encapsulated by algae membrane, no fluorescence was detected, and the carapace morphology remained intact after ingesting algae for 80 min, in contrast to the rapid carapace deformation by direct Hg2+ absorption. The D. carinata showed higher mortality by direct Hg2+ immersion than via food ingestion. The reason for the high mortality after Hg2+ immersion was possibly due to carapace deformity after chemical reaction between chitin and mercury, but the biochemical pathway leading to morality needs further study.

The complete mitochondrial DNA genome of Chinese Daphnia carinata (Clasocera: Daphniidae).

In this paper, we determined the complete mitochondrial genome of Chinese Daphnia carinata for the first time by the long and accurate polymerase chain reaction and primer-walking methods. It was 15,245 bp in length, with an A + T content of 70.35%, containing 37 typical animal mitochondrial genes and an A + T-rich region. The COI gene started with ACTA. All the 22 typical tRNA genes had a classical cloverleaf structure except for trnS1, in which the D-stem pairings in the DHU arm were absent.

Cloning, expression and localization of DacaCSP2 and DacaCSP3 during different reproductive stages in Daphnia carinata.

Daphnia carinata are unique freshwater crustaceans that undergo both sexual and asexual reproduction, depending on environmental factors. While the molecular mechanism behind the reproductive transformation has been unknown, chemosensory proteins (CSPs) may be involved. We have cloned the cDNA sequences of two CSP genes from D. carinata using primers based on homologous sequences and rapid amplification of cDNA ends (RACE). The full-length cDNA of DacaCSP2 (GenBank accession no: KM624608) was 632 bp, with an ORF (open reading frame) of 330 bp encoding a 12.02 kDa protein; and the full-length cDNA of DacaCSP3 (GenBank accession no: KM624609) was 935 bp, with an ORF of 342 bp encoding a 12.78kDa protein. Both CSPs encoded an N-terminal signal peptide, four conserved cysteines, an OS-D superfamily domain, a 2Fe-2S ferredoxin domain, an anaphylatoxin domain and an EGF-like domain. DaCaCSP2 and DaCaCSP3 proteins were most closely related to CSPs from Daphnia pulex and were more distantly related to CSPs from other insects. Using quantitative PCR, we found expression levels of DaCaCSP2 and DaCaCSP3 mRNA were highest in sexual females, followed by parthenogenetic females, and lowest in males. The expression levels of DaCaCSP2 and DaCaCSP3 mRNA also increased at lower temperatures, which suggested they could respond to environmental cues. Whole mount in situ hybridization (ISH) showed that DaCaCSP2 and DaCaCSP3 were expressed mainly in the ovaries, summer eggs, thoracic limbs, rectum and second antennae in sexual females; while they were expressed mainly in the ovaries, thoracic limbs, rectum and second antennae in parthenogenetic females. Together, these results suggest that DacaCSP2 and DacaCSP3 may respond to environmental cues and control the reproductive switch from sexual to asexual reproduction in D. carinata.

Cloning, expression and cellular localization of the Doublesex gene in the water flea, Daphnia carinata, during different developmental stages.

In this study, one of Doublesex genes from the common freshwater cladoceran Daphnia carinata, designated DapcaDsx1, was cloned using primers based on homologous sequences and rapid amplification of cDNA ends (RACE). qPCR was employed to quantify differences in DapcaDsx1 expression between the different sexual phases, with expression levels being higher in sexual females. The role of DapcaDsx1 in the reproductive transformation was further investigated in parthenogenetic-phase females and sexual-phase females using whole-mount in situ hybridization. This cellular localization study showed specific expression of DapcaDsx1 in the thoracic segments, second antenna and part of the ventral carapace. Higher expression levels were exhibited in sexual females compared to parthenogenetic females. This suggests that the DapcaDsx1 gene plays significant roles in switching modes of reproduction and during sexual differentiation.