Marine Copepod Tigriopus Japonicus Research Articles

Microplastics at an environmentally relevant dose enhance mercury toxicity in a marine copepod under multigenerational exposure: Multi-omics perspective

Here, we subjected the marine copepod Tigriopus japonicus to environmentally-relevant concentrations of microplastics (MPs) and mercury (Hg) for three generations (F0–F2) to investigate their physiological and molecular responses. Hg accumulation and phenotypic traits were measured in each generation, with multi-omics analysis conducted in F2. The results showed that MPs insignificantly impacted the copepod’s development and reproduction, however, which were significantly compromised by Hg exposure. Interestingly, MPs significantly increased Hg accumulation and consequently aggravated this metal toxicity in T. japonicus, demonstrating their carrier role. Multi-omics analysis indicated that Hg pollution produced numerous toxic events, e.g., induction of apoptosis, damage to cell/organ morphogenesis, and disordered energy metabolism, ultimately resulting in retarded development and decreased fecundity. Importantly, MPs enhanced Hg toxicity mainly via increased oxidative apoptosis, compromised cell/organ morphogenesis, and energy depletion. Additionally, phosphoproteomic analysis revealed extensive regulation of the above processes, and also impaired neuron activity under combined MPs and Hg exposure. These alterations adversely affected development and reproduction of T. japonicus. Overall, our findings should offer novel molecular insights into the response of T. japonicus to long-term exposure to MPs and Hg, with a particular emphasis on the carrier role of MPs on Hg toxicity.

Elevated temperature as a dominant driver to aggravate cadmium toxicity: Investigations through toxicokinetics and omics

Despite the great interest in the consequences of global change stressors on marine organisms, their interactive effects on cadmium (Cd) bioaccumulation/biotoxicity are very poorly explored, particularly in combination with the toxicokinetic model and molecular mechanism. According to the projections for 2100, this study investigated the impact of elevated pCO2 and increased temperature (isolated or joint) on Cd uptake dynamics and transcriptomic response in the marine copepod Tigriopus japonicus. Toxicokinetic results showed significantly higher Cd uptake in copepods under increased temperature and its combination with elevated pCO2 relative to the ambient condition, linking to enhanced Cd bioaccumulation. Transcriptome analysis revealed that, under increased temperature and its combination with elevated pCO2, up-regulated expression of Cd uptake-related genes but down-regulation of Cd exclusion-related genes might cause increased cellular Cd level, which not only activated detoxification and stress response but also induced oxidative stress and concomitant apoptosis, demonstrating aggravated Cd biotoxicity. However, these were less pronouncedly affected by elevated pCO2 exposure. Therefore, temperature seems to be a primary factor in increasing Cd accumulation and its toxicity in the future ocean. Our findings suggest that we should refocus the interactive effects between climate change stressors and Cd pollution, especially considering temperature as a dominant driver.

Seawater warming intensifies nickel toxicity to a marine copepod: a multigenerational perspective

Due to human activities, marine organisms are frequently co-stressed with nickel (Ni) pollution and seawater warming; nevertheless, very scarce information is known about their interaction in marine biota under a multigenerational scenario. Here, after verifying the interaction of Ni and warming via a 48-h acute test, we conducted a multigenerational experiment (F0-F2), in which the marine copepod Tigriopus japonicus was exposed to Ni at environmentally realistic concentrations (0, 2, and 20 µg/L) under ambient (22℃) and predicted seawater warming (26℃) conditions. Ni accumulation and the important life history traits were analyzed for each generation. Results showed that Ni exposure caused Ni bioaccumulation and thus compromised the survivorship and egg production of T. japonicus. In particular, seawater warming significantly increased Ni accumulation, thus intensifying the negative effects of Ni on its survivorship and development. Overall, this study suggests that Ni multigenerational exposure even at environmentally realistic concentrations could produce a significant impact on marine copepod's health, and this impact would be intensified under the projected seawater warming, providing a mechanistic understanding of the interaction between warming and Ni pollution in marine organisms from a multigenerational perspective.

Multigenerational effects of arsenate on development and reproduction in marine copepod Tigriopus japonicus

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.

High and diurnally fluctuating carbon dioxide exposure produces lower mercury toxicity in a marine copepod

Coastal waters have experienced fluctuations in partial pressure of carbon dioxide (pCO2) and mercury (Hg) pollution, yet little is known concerning how natural pCO2 fluctuations affect Hg biotoxicity. Here, a marine copepod Tigriopus japonicus was interactively exposed to different seawater pCO2 (ambient 400, steady elevated 1000, and fluctuating elevated 1000 ± 600 μatm) scenarios and Hg (control, 2 μg/L) treatments for 7 d. The results showed that elevated pCO2 decreased Hg bioaccumulation, and it was even more under fluctuating elevated pCO2 condition. We found energy depletion and oxidative stress under Hg-treated copepods, while combined exposure initiated compensatory response to alleviate Hg toxicity. Intriguingly, fluctuating acidification presented more immune defense related genes/processes in Hg-treated copepods when compared to steady acidification, probably linking with the greater decrease in Hg bioaccumulation. Collectively, understanding how fluctuating acidification interacts with Hg contaminant will become more crucial in predicting their risks to coastal biota and ecosystems.

Joint effects of temperature and copper exposure on developmental and gene-expression responses of the marine copepod Tigriopus japonicus.

There is growing contamination of copper (Cu) in the marine environment, particularly after the ban of organotin compounds and the increase of the use of Cu-based antifouling paints. Although there are increasing research interests in temperature-dependent chemical toxicity to aquatic organisms, most existing studies focused on acute impacts of chemicals at high concentrations. This study aimed to investigate the interacting effect of temperature and copper exposure at environmentally relevant concentrations on survival and development in the marine copepod Tigriopus japonicus with a partial life-cycle toxicity test. Expressions of five stress response genes in the copepod, namely two glutathione S-transferases (GST-S and GST-O), two heat shock proteins (HSP70 and HSP90), and glutathione reductase (GR) were also investigated. The copepod's survival was significantly impaired at 15 °C after development to adult stage, while its developmental time reduced significantly with increasing temperature. Copper at the two environmentally relevant test concentrations had no significant impacts on these apical endpoints whereas the interaction between Cu and temperature was more significant in modulating gene expressions. GST-S, GST-O and HSP90 genes in copepods exposed to 100 µg Cu L-1 were significantly upregulated at 20 °C. At 32 °C, most genes were either insignificantly expressed or down-regulated, compared to the control, likely suggesting that thermal stress inhibited the copepod's antioxidative defense system. Overall, the results revealed that the joint Cu and thermal stresses have significantly elicited antioxidative system in the copepods. It clearly demonstrated the need for more fundamental studies about potential impacts of different environmental factors such as temperature on chemical toxicity under realistic scenario of marine pollution.

Nanoplastics pose a greater effect than microplastics in enhancing mercury toxicity to marine copepods

Due to human activities, high abundances of nano/microplastics (N/MPs) concurrent with metal pollution have become a serious problem in the global marine environment. Because of displaying a high surface-area-to-volume ratio, N/MPs can serve as the carriers of metals and thus increase their accumulation/toxicity in marine biota. As one of the most toxic metals, mercury (Hg) causes adverse effects on marine organisms but whether environmentally relevant N/MPs can play a vector role of this metal in marine biota, as well as their interaction, is poorly known. To evaluate the vector role of N/MPs in Hg toxicity, we first performed the adsorption kinetics and isotherms of N/MPs and Hg in seawater, as well as ingestion/egestion of N/MPs by marine copepod Tigriopus japonicus, and second, the copepod T. japonicus was exposed to polystyrene (PS) N/MPs (500-nm, 6-μm) and Hg in isolation, combined, and incubated forms at environmentally relevant concentrations for 48 h. Also, the physiological and defense performance including antioxidant response, detoxification/stress, energy metabolism, and development-related genes were assessed after exposure. The results indicated N/MPs significantly increased Hg accumulation and thus its toxicity effects in T. japonicus as exemplified by decreased transcription of genes related to development and energy metabolism and increased transcriptional levels of genes functioning in antioxidant and detoxification/stress defense. More importantly, NPs were superimposed onto MPs and produced the most vector effect in Hg toxicity to T. japonicus, especially in the incubated forms. Overall, this study highlighted the role of N/MPs as a potential risk factor for increasing the adverse effects of Hg pollution, and emphasized the adsorption forms of contaminants by N/MPs should doubly be considered in the continuing researches.

Physiological and Molecular Responses of a Marine Copepod Under Multigenerational Exposure to Coastal Warming and Mercury at an Environmentally Realistic Concentration

In this work, we investigated how a warmer temperature (26 °C) and mercury pollution (1 μg/L) affect the marine copepod Tigriopus japonicus across three generations (F0–F2). Several phenotypic traits and mercury accumulation were measured in each generation, and the proteome of the F2 copepods was analyzed. The results show that a warmer temperature and mercury exposure significantly affected the phenotypic traits. Combined exposure significantly increased mercury bioaccumulation and consequently potentiated its toxicity in each generation relative to mercury treatment alone. The proteomic analysis demonstrated that the warmer temperature caused the copepods to upregulate the determination of adult lifespan pathway to maintain rapid growth but at the cost of an organism’s fitness, as exemplified by the male-biased sex ratio and compromised reproduction. Mercury pollution had several toxic effects, e.g., energy depletion and reduced defense performance, that ultimately translated to decreased reproduction. Importantly, the warmer temperature aggravated mercury toxicity, mainly by disturbing energy metabolism and impairing detoxification systems, which were linked to depressed feeding and reproduction in T. japonicus. Our study provides a mechanistic, multigenerational understanding of the response of T. japonicus to coastal warming and mercury pollution, with emphasis on enhanced mercury toxicity in marine copepods under warmer conditions.

Herbicide prometryn adversely affects the development and reproduction of Tigriopus japonicus by disturbing the ecdysone signal pathway and chitin metabolic pathway

Prometryn, a widely used triazine herbicide in agriculture and aquaculture, has been commonly detected in marine environments, but its effects on the marine copepod are unknown. In this study, marine copepod Tigriopus japonicus was chronically exposed to environmentally relevant concentrations of prometryn to investigate its impacts and potential mechanism of action. The results showed that 0.5, 5, and 50 μg/L prometryn delayed the first spawning time and hatching time, reduced the fecundity, and inhibited the population growth rate. Moreover, exposure to 0.5, 5 and 50 μg/L prometryn decreased food ingestion, the content of C and N elements, nutrient accumulation and body size, but increased the content of 20-hydroxyecdysone (20E). Transcriptome analysis showed that 50 μg/L prometryn down-regulated 1431 genes, which were mainly enriched in lysosome pathway and chitin binding and cuticle construction process. The results of qRT-PCR showed that the expression of key genes involved in juvenile hormone synthesis and chitin metabolic pathways were also inhibited after prometryn exposure. Molecular docking revealed that prometryn could bind to ecdysone receptor (EcR) and UDP-N-acetylglucosamine pyrophosphorylase (UAP), components of the ecdysteroid nuclear receptor complex. Therefore, environmental relevant prometryn delayed the molting and development of T. japonicus by disrupting the ecdysone signal pathway and chitin metabolic pathway through binding to EcR and UAP. This study provides new insights into toxic effects and molecular mechanisms of prometryn on marine copepods.

Reduced Fitness and Elevated Oxidative Stress in the Marine Copepod Tigriopus japonicus Exposed to the Toxic Dinoflagellate Karenia mikimotoi.

Blooms of the toxic dinoflagellate Karenia mikimotoi cause devastation to marine life, including declines of fitness and population recruitment. However, little is known about the effects of them on benthic copepods. Here, we assessed the acute and chronic effects of K. mikimotoi on the marine benthic copepod Tigriopus japonicus. Results showed that adult females maintained high survival (>85%) throughout 14-d incubation, but time-dependent reduction of survival was detected in the highest K. mikimotoi concentration, and nauplii and copepodites were more vulnerable compared to adults. Ingestion of K. mikimotoi depressed the grazing of copepods but significantly induced the generation of reactive oxygen species (ROS), total antioxidant capacity, activities of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase), and acetylcholinesterase. Under sublethal concentrations for two generations, K. mikimotoi reduced the fitness of copepods by prolonging development time and decreasing successful development rate, egg production, and the number of clutches. Our findings suggest that the bloom of K. mikimotoi may threaten copepod population recruitment, and its adverse effects are associated with oxidative stress.

Bioaccumulation of silver nanoparticles in marine copepod Tigriopus japonicus under waterborne and dietary exposure

At present, the emission of silver nanoparticles (AgNPs) will affect a variety of aquatic organisms. AgNPs may be transferred to the food web through the bioaccumulation or biomagnification of the food chain, which is harmful to at higher trophic level organisms, and even threatens human health. Therefore, in this study, we tested the acute toxicity effect of AgNPs to marine copepod Tigriopus japonicus, and analyzed the Ag bioaccumulation in T. japonicus under short-term waterborne (exposure of AgNPs to T. japonicus) and dietary exposure (AgNP-accumulated algae cells of Isochrysis galbana are used to feed T. japonicus). The results showed that, after 48 h exposure, significant effects of AgNPs on the mortalities were observed (LC50=6.330 mg·L−1). The bioaccumulation of Ag by T. japonicus under different exposure modes was positively correlated with exposure concentration and time, and the bioaccumulation level under waterborne exposure was significantly higher. The biomagnification of T. japonicus was not observed under 24 h dietary exposure of different AgNP concentrations (BMFs<1), but the transfer efficiency was higher under low exposure concentration (BMFlow concentration>BMFhigh concentration). This study is helpful to understand the toxicity mechanism and effect of AgNPs on aquatic organisms.

Transcriptional and toxic responses to saxitoxin exposure in the marine copepod Tigriopus japonicus

Saxitoxin (STX) is a highly toxic marine neurotoxin produced by phytoplankton and a growing threat to ecosystems worldwide due to the spread of toxic algae. Although STX is an established sodium channel blocker, the overall profile of transcriptional levels in STX-exposed organisms has yet to be described. Here, we describe a toxicity assay and transcriptome analysis of the copepod Tigriopus japonicus exposed to STX. The half-maximal lethal concentration of STX was 12.35 μM, and a rapid mortality slope was evident at concentrations between 12 and 13 μM. STX induced changes in swimming behavior among the copepods after 10 min of exposure. In transcriptome analysis, gene ontology revealed that the genes involved in nervous system and gene expression were highly enriched. In addition, the congenital neurological disorder and nuclear factor erythroid 2-related factor 2–mediated oxidative stress pathways were identified to be the most significant in network analysis and toxicity pathway analysis, respectively. This study provides valuable information about the effects of STX and related transcriptional responses in T. japonicus.

Chronic effects of nano and microplastics on reproduction and development of marine copepod Tigriopus japonicus

This study aimed to examine the impact of chronic (30 days) exposure to polystyrene microplastics (PS-MPs) of different sizes (50 nm and 2 µm) and at different concentrations (0.5 μg/L and 100 mg/L) to marine copepod Tigriopus japonicus. Polystyrene microplastics affected survival rates in size- and concentration-dependent manners. The LC50s values of 50 nm and 2 µm PS-MPs were 0.10 mg/L and 3.92 mg/L, respectively. The developmental time was delayed by 50 nm PS-MPs, and Usp expression was downregulated. Reproduction was negatively affected by 2 µm PS-MPs even at environmentally relevant concentrations; however, the expression of Vtg was not altered. The production rates of reactive oxygen species and nitric oxide also increased after exposure to PS-MPs; but this effect was independent of particle size. The expression levels of Cat and Tnf, genes related to oxidative stress and inflammation, respectively, were upregulated by exposure to PS-MPs, independently of particle size. Meanwhile, the level of oxidative stress in T. japonicus was not significantly affected by PS-MPs at environmentally relevant concentrations. This study suggests that nano-sized PS-MPs are not always more toxic than micro-sized PS-MPs, and that oxidative stress is a key factor in determining the toxic effect on T. japonicus at high concentrations.

Effects of Leachates from Hydrothermal Ore Particulates on Life Cycle Parameters and Expression of Defense-Related Genes in the Marine Copepod Tigriopus japonicus

Effects of Leachates from Hydrothermal Ore Particulates on Life Cycle Parameters and Expression of Defense-Related Genes in the Marine Copepod Tigriopus japonicus

Evaluation of ecotoxicological effects associated with coastal sediments of the Yellow Sea large marine ecosystem using the marine copepod Tigriopus japonicus

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.

Effects of particulate matter (PM2.5) on life history traits, oxidative stress, and defensome system in the marine copepod Tigriopus japonicus

Particulate matter (PM2.5) generated in large cities creates new problems in marine ecosystems and may adversely affect its inhabitants. However, the mechanisms underlying the same remain unclear; hence, we investigated the effects of PM2.5 on life history traits (e.g., mortality, development, and fecundity), cellular reactive oxygen species (ROS) levels, antioxidant enzyme (e.g., glutathione peroxidase [GPx], superoxide dismutase [SOD], and catalase [CAT]) activities, and the transcript levels of detoxification-related genes (cytochrome P450s [CYPs]) and antioxidant (glutathione S-transferases [GSTs]) in the copepod Tigriopus japonicus. Among the life history traits, developmental time was the only trait to significantly deviate (P < 0.05) in response to PM2.5 (compared to that in the controls). Significant changes in ROS levels and antioxidant enzymatic activities (P < 0.05) in response to PM2.5, suggested that PM2.5 can induce oxidative stress, leading to adverse effects on the T. japonicus life history. In addition, PM2.5 induced a differential regulation of various CYP and GST genes, particularly CYP307E1, GST-kappa, and GST-sigma were significantly upregulated (P < 0.05), suggesting that these genes likely play crucial roles in detoxification mechanisms and could be useful as reliable biomarkers for PM2.5 toxicity. Overall, the results of this study provide new insights into the potential toxicity of PM2.5.

Acute toxicity of tire wear particles, leachates and toxicity identification evaluation of leachates to the marine copepod, Tigriopus japonicus

Tire wear particles (TWPs) have been characterized as microplastics in recent years, and many of these TWPs will be eventually deposited in coastal areas, leading to adverse effects to marine organisms. Results of the acute toxicity test in this study showed that the 96-h LC50 values of the particles and leachate were 771.4 mg/L (95% CI = 684.4–869.6 mg/L) and 5.34 g/L (95% CI = 4.75–6.07 g/L), respectively. The chemical constituents of TWP and the leachate are very complex, and little research has been conducted to determine which of these constituents contribute to the toxicity of TWP leachate to marine organisms. Therefore, the composition of the TWP and leachate was analyzed, and a variety of chemicals were identified, including metals (Mn, Zn, etc.) and organic compounds (cyclohexanthiol, 4-ethyl-1,2-dimethylbenzene, benzothiazole, stearic acid, N-(1,3-dimethylbutyl)-N′-phenyl-p-phenylenediamine, etc.). In addition, the marine copepod Tigriopus japonicus was applied as a model species in the toxicity identification evaluation study to characterize, identify and confirm the toxicity-causing substances in the TWP leachate. Zn was identified and confirmed as the main toxicant contributing to the toxicity. Furthermore, Zn concentrations in the leachate over time were investigated. The release of Zn from TWPs to the aquatic environment was slow, and conformed to a parabolic model with a release constant k of 2.06. The organic component, benzothiazole, exhibited an antagonistic effect with zinc in the acute toxicity of the TWP leachate.

CO2-driven seawater acidification increases cadmium toxicity in a marine copepod

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.

Release of Microplastics from Discarded Surgical Masks and Their Adverse Impacts on the Marine Copepod Tigriopus japonicus

Surgical masks (SMs) are the most commonly used personal protective equipment during the COVID-19 pandemic. Due to their vast use and inappropriate disposal worldwide, SMs could potentially cause serious microplastic (MP) pollution in coastal marine environments. This study aimed to investigate the kinetic release of MPs from polypropylene SMs (PP-SMs) in seawater and to evaluate the chronic toxicity of the released MPs to the marine copepod Tigriopus japonicus. On the basis of the results of our kinetic study and available relevant data, we estimated that SMs discarded throughout the year 2020 would lead to >1370 trillion MPs entering the coastal marine environment globally, with a release rate of 396 billion MPs per day. Our results also demonstrated that the copepods ingested the MPs released from PP-SMs, causing a significant decline in their fecundity. The results clearly suggest the MPs released from improperly discarded SMs could have a long-term domino effect on coastal marine ecosystems. To minimize the risk of this emerging threat, better environmental management, policy, and law enforcement for ensuring the proper disposal of SMs are deemed to be necessary.

Predicted Near-Future Oceanic Warming Enhances Mercury Toxicity in Marine Copepods.

The effects of acute mercury exposure (118µg/L) on the marine copepod Tigriopus japonicus were examined at 22 and 25°C for 24h and compared with controls. Mercury accumulation and seven genes related to antioxidant/stress responses were analyzed after exposure. The 24-h LC50 value decreased in the warmer environment and mercury accumulation was elevated. Under both temperatures, mercury significantly affected the expression of all analyzed genes and probably caused oxidative stress. Intriguingly, at the same mercury concentration, most genes were upregulated at the higher relative to the lower temperature, and the copepods likely initiated more compensatory reactions to counteract increased mercury toxicity associated with the warmer temperature. Overall, this study suggests a molecular mechanism by which marine copepods could respond to future oceanic warming and mercury pollution.