Arsenic (As) can be transferred along the food chain, while little is known about the toxic effects of dietborne As on marine copepods. In this study, we investigated the short-term and long-term effects of waterborne and dietborne As exposure on the bioaccumulation and biotransformation, as well as developmental toxicity of Tigriopus japonicus. Under acute As exposure, As bioaccumulation increased and reached a plateau with increasing exposure concentration. Moreover, As accumulation at dietborne exposure was 4.3 and 5.7 times greater than that at control group for AsIII and AsV, respectively. At chronic As exposure, As accumulation increased continuously with exposure time, with a 2.8-day extension of development time and a 45% reduction in 10-d fecundity under dietborne exposure compared to control, whereas 2.3-day extension of development time and a 20% reduction in 10-d fecundity were observed under waterborne exposure. Among As species, inorganic As had the highest concentrations, but the proportion of inorganic As decreased from 89% to 63% during 4 to 21 d of exposure, suggesting the conversion of inorganic As to organic As. The organic As was dominated by arsenobetaine (AsB, 13-25%), followed by monomethylarsenic (MMA, 8-25%). These results suggest that dietborne exposure has more pronounced toxic effects on T. japonicus, but the toxicity of As could be reduced through biotransformation under chronic exposure. Therefore, the arsenic species should be considered when assessing As toxicity.

Arsenic (As) is a persistent toxic substance, however, its toxicity to marine zooplankton remains unclear. In this study, copepods were exposed to a series of dissolved arsenate (As(V)) for four generations (F0-F3) and subsequently depurated in clean seawater for two generations (F4-F5) to assess multigenerational toxicity of As(V). As(V) exposure prolonged copepod development. The development time were 1.9, 2.4, and 3.4 days longer than the control in F0 when exposed to 50, 100, and 500 μg/L As(V), respectively, and the toxicity increased with generations. Moreover, As(V) reduced the reproductive capacity of copepods, and this effect become more severe during generation succession. The 10-day fecundities were reduced from 80 to 85 eggs per female in the control to 42 eggs per female, the lowest level, in 500 μg/L As(V) exposure group in F3. Nevertheless, the fecundity was recovered to the control level in the offspring of the 50 and 100 μg/L As(V) exposed groups (F4), suggesting it was an acclimation effect of copepods during As(V) exposure. In addition, the survival rate, development time, and reproductive parameters were significantly correlated with the As accumulation in copepods. Overall, As(V) exposure caused As bioaccumulation which negatively affected copepods\' survival, development, and reproductive traits, and this toxic effect was amplified with generations and concentrations. Therefore, the multigenerational toxicity of As should be considered in the environmental risk assessments.

The use of beneficial microbes, i.e., probiotics, to reduce pathogens and promote the performance of the target species is an important management strategy in mariculture. This study aimed to investigate the potential of four microbes, Debaryomyces hansenii, Ruegeria mobilis, Lactobacillus plantarum, and Bacillus subtilis, to suppress Vibrio and increase survival, population growth and digestive enzyme activity (protease, lipase, and amylase) in the harpacticoid copepod, Tigriopus japonicus. Copepod, T. japonicus stock culture with an initial mean density of 50 individual/mL (25 adult male and 25 adult female) was distributed into five treatments (i.e., four experimental and a control, each with four replicates; repeated twice) using 20 beakers (100 mL capacity each). The copepods were fed a mixture of the dinoflagellate Alexandrium tamarense and the diatom Phyaeodactylum tricornutum (3 × 104 cells/mL-1). Each microbe\'s concentration was adjusted at 108 CFU/mL-1 and applied to the culture condition. D. hansenii, L. plantarum, and B. subtilis all improved the copepods\' survival and population growth, likely by including a higher lipase activity (P < 0.05). In contrast, using R. mobilis did not improve the copepod\'s culture performance compared to control. B. subtilis was the most effective in decreasing the copepod\'s external and internal Vibrio loading. The probiotic concentrations in the copepod decreased within days during starvation, suggesting that routine re-application of the probiotics would be needed to sustain the microbial populations and the benefits they provide. Our results demonstrated that D. hansenii and B. subtilis are promising probiotics for mass copepod culture as live food for mariculture purposes.

Toxicology is not only for eco-risk assessments, but also for the real-time environmental monitoring based on the quick response of specific biomarkers. Ferritin gene (ftn) is a potential biomarker involving in crucial protective responses in biota. However, little information is available concerning the ftn in marine copepod Acartia tonsa (A. tonsa), a model organism widely applied in toxicology assessments. Our study for the first time identified and characterized the ftn in A. tonsa, along with its time-dependent transcriptional response to the reproductive toxicity of two newly emerged nanomaterials. The full-length cDNA of ftn contains a 114-bp 5\'-untranslated region (UTR), a 236-bp 3\'-untranslated region, and a 510-bp open reading frame which encodes an 18.51 kDa polypeptide composed of 169 amino acids. The ftn sequence has an iron binding signature and a potential phosphorylation site, which is closely-related to the ftn of Calanus sinicus and Pseudodiaptomus annandalei genes at the phylogenetical level. The ftn showed a quick and highly sensitive response to nanomaterial exposures, even at no observed effect concentrations. In detail, after exposure to nickel nanomaterials (up to 17.0 mg/L), the ftn was significantly upregulated immediately at 0.5 h and peaked at 9.5-fold in adults within 48 h, along with a significant reduction of egg hatching rate. When exposed to CdSe/ZnS quantum dots (up to 135 mg/L), no significant change in egg productions or hatching rates was observed, while the expression of ftn still significantly increased to over 3.0-fold in the initial 48 h. After that, the upregulation of ftn induced by CdSe/ZnS quantum dots or nickel nanoparticles both gradually returned back within 96 h. These findings demonstrate the highly sensitive response of this new cloned ftn to nanomaterial exposures, and highlight the suitability of ftn in A. tonsa as a promising biomonitor for nano-contamination in marine environments.

There is increasing concern about the adverse impact of exposure to microplastic, as an emerging pollutant, on wild organisms, and particularly on organisms co-exposed to microplastic and other environmental contaminants. It has been widely reported that the combination of microplastics and heavy metals showed obvious toxicity to organisms in terms their growth and development. The present study was performed to determine the impact of binary metal mixtures of cadmium (Cd) and polystyrene microplastic (PS-microplastic) on Tigriopus japonicus, a typical marine model organism, using a titration design. Increasing concentrations of PS-microplastic (2 μg/L, 20 μg/L, and 200 μg/L) were titrated against a constant concentration of Cd (15.2 μg/L). The results showed no significant impact of exposure to this dose of Cd or co-exposure to Cd and the lowest dose of PS-microplastic examined (2 μg/L). However, the feeding rate, filtration rate, oxygen consumption rate, and hatching number declined significantly in T. japonicus co-exposed to Cd and higher concentrations of PS-microplastic (20 μg/L and 200 μg/L) (p < 0.05). Furthermore, the development of F1 larvae from nauplius stage (N) to adult stage (A) was markedly delayed when co-exposed to Cd and higher doses of PS-microplastic (20 and 200 μg/L), and the effects persisted to the F2 larval stage. Interestingly, the present titration design did not affect the sex ratio or number of oocysts in either the F1 or F2 generation. These results indicated that the current marine environmental concentrations of Cd and microplastic are safe for wild organisms. Further studies are required to address the knowledge gap regarding toxicological effects at the cellular and molecular levels.

Coastal systems experience diel fluctuation of pCO2 and cadmium (Cd) pollution; nevertheless, the effect of fluctuating pCO2 on Cd biotoxicity is poorly known. In this study, we initially performed the isotopically enriched organism bioassay to label Tigriopus japonicus with 113Cd (5 μg/L) to determine the Cd accumulation rate constant (kaccu) under ambient (400 μatm) and steadily (1000 μatm) and fluctuatingly elevated (1000 ± 600 μatm) pCO2 conditions for 48 h. Next, T. japonicus was interactively subjected to the above pCO2 exposures at Cd (control, 5, and 500 μg/L) treatments for 7 d. Biochemical and physiological responses for copepods were analyzed. The results showed that steadily increased pCO2 facilitated Cd bioaccumulation compared to ambient pCO2, and it was more under fluctuating acidification conditions. Despite compensatory reactions (e.g., increased energy production), Cd ultimately induced oxidative damage and apoptosis. Meanwhile, combined treatment exhibited higher toxicity (e.g., increased apoptosis) relative to Cd exposure, and even more if fluctuating acidification was considered. Intriguingly, fluctuating acidification inhibited Cd exclusion in Cd-treated copepods compared to steady acidification, linking to higher Cd kaccu and bioaccumulation. Collectively, CO2-driven acidification could aggravate Cd toxicity, providing a mechanistic understanding of the interaction between seawater acidification and Cd pollution in marine copepods.

A copepod bioassay with Tigriopus japonicus was applied to evaluate the relative ecotoxicity of sediments in the Yellow and Bohai seas, and contributions of individual PAHs to copepod toxicity were evaluated. Mean toxicity was greatest in the Yellow Sea of China, followed by the Bohai Sea and Yellow Sea of Korea. Elevated concentrations of sedimentary PAHs, alkylphenols, and styrene oligomers back-supported the significant toxicities observed in bioassay. Copepod toxicity in relation to PAHs indicated the greatest contribution by indeno[1,2,3-c,d]pyrene. However, lacked contribution by PAHs, viz., 2.4 and 3.0 % for the total immobilization and mortality, respectively, indicated a large proportion of unknown toxicants being widely distributed along the Yellow Sea Large Marine Ecosystem (YSLME) coastline. Overall, the present study provides useful baseline information for evaluating the potential sedimentary toxicants, with emphasizing further investigation to identify the unknown toxicants at an LME scale, and elsewhere.

Here, we examined the 48-h acute toxicity of cadmium (Cd) in the marine copepod Tigriopus japonicus under two pCO2 concentrations (400 and 1000 μatm). Subsequently, T. japonicus was interactively exposed to different pCO2 (400, 1000 μatm) and Cd (control, 500 μg/L) treatments for 48 h. After exposure, biochemical and physiological responses were analyzed for the copepods. The results showed that the 48-h LC50 values of Cd were calculated as 12.03 mg/L and 9.08 mg/L in T. japonicus, respectively, under 400 and 1000 μatm pCO2 conditions. Cd exposure significantly promoted Cd exclusion/glycolysis, detoxification/stress response, and oxidative stress/apoptosis while it depressed that of antioxidant capacity. Intriguingly, CO2-driven acidification enhanced Cd bioaccumulation and its toxicity in T. japonicus. Overall, our study provides a mechanistic understanding about the interaction between seawater acidification and Cd pollution in marine copepods.

Hypoxia is intensified in tropical and subtropical estuarine and coastal waters and brings about lethal and sublethal effects to marine copepods. The physiological and behavioral responses of the subtropical calanoid copepod Temora turbinata were tested after short-term exposure to hypoxia. The LD50 values were 3.02 ± 0.21, 2.00 ± 0.35, and 3.11 ± 0.31 mg L-1 for nauplii (II-III), copepodites (II-III), and female adults, respectively. With a decrease in the ambient dissolved oxygen (DO) level from 8 to 0.5 mg L-1, the ingestion rates decreased significantly at all life stages, as did oxygen consumption in female adults. In an artificial stratification column with a DO gradient, female adults exhibited an obvious avoidance response to the hypoxic bottom layer. Our study provides preliminary evidence for high hypoxia sensitivity in T. turbinata and implies that the DO level may be the main factor controlling the distribution of this species in tropical and subtropical coastal and estuarine waters.

The effects of microplastics pollution on the marine ecosystem have aroused attention. Copepod grazing stimulates dimethylsulfide (DMS) release from dimethylsulfoniopropionate (DMSP) in phytoplankton, but the effect of microplastics exposure on DMS and DMSP production during copepod feeding has not yet been revealed. Here, we investigated the effects of polyethylene (PE) and polyamide-nylon 6 (PA 6) microplastics on ecotoxicity and DMS/DMSP production in the copepod Tigriopus japonicus. The microplastics had detrimental effects on feeding, egestion, reproduction, survival, and DMS and DMSP production in T. japonicus and presented significant dose-response relationships. The 24 h-EC50 for ingestion rates (IRs) of female T. japonicus exposed to PE and PA 6 were 57.6 and 58.9 mg L-1, respectively. In comparison, the body size of the copepods was not significantly affected by the microplastics during one generation of culture. Ingesting fluorescently labeled microplastics confirmed that microplastics were ingested by T. japonicus and adhered to the organs of the body surface. T. japonicus grazing promoted DMS release originating from degradation of DMSP in algal cells. Grazing-activated DMS production decreased because of reduced IR in the presence of microplastics. These results provide new insight into the biogeochemical cycle of sulfur during feeding in copepods exposed to microplastics.