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Chronic cerebral hypoperfusion (CCH) is the main characteristic of an aged brain showing cerebrovascular alterations. Our previous study that the morphological changes in the pial arteries accompany a decrease in the cerebral blood flow in aged mouse brains, and it raises the question of whether artificial CCH can induce the same changes in brain vessel morphology. Here, we examined the effect of CCH on cerebrovascular morphology. Using a microcoil-induced chronic cerebral hypoperfusion (MCH) model, we induced CCH for 8 and 12 weeks. The cerebrovasculature morphology was evaluated using three-dimensional vessel analysis and compared with that of aging mice. We found the morphological changes in brain vessels of MCH mice, indicating that the CCH can induce cerebrovascular alteration. However, the changes in brain vessel morphology in the MCH mice were different in detail from those in the aging mice. Aging mice showed an increase in vessel tortuosity and thinned string vessels; MCH mice mainly showed thinned string vessels. This suggests that CCH may be a cause of age-related cerebrovascular alterations. • Chronic cerebral hypoperfusion (CCH) causes changes in cerebrovascular morphology. • We found morphological differences in cerebrovasculature between CCH and aged mice. • CCH is a possible cause of age-related cerebrovascular changes. [ABSTRACT FROM AUTHOR]

• Exposure of OA to copepod resulted in reduction in the growth rate with decreased fecundity and body length. • GST and GR activities were increased in response to OA-induced oxidative stress. • GPx and SOD were decreased in a pH-dependent manner. • GSTs2b was significantly up-regulated in response to OA. Ocean acidification (OA) is caused by alteration of global ocean carbon chemistry due to the increased pCO 2 in the atmosphere and caused deleterious impacts on the marine ecosystem. Although various detrimental effects of OA were reported in marine organisms, the potential impact of OA on aquatic invertebrates still remains largely unknown. Here, we examined changes in life parameters and antioxidant system in response to low pH (7.5 and 7) in the marine copepod Tigriopus japonicus. Exposures to lower pHs (pH 7.5 and 7.0) of copepods resulted in lengthening of the developmental time with decreased fecundity and body length. Also, they showed increased reactive oxygen species contents with enhanced glutathione S -transferase and glutathione reductase activities but decreased glutathione peroxidase and superoxide dismutase activities in pH-dependent manner, indicating that OA exposure caused disturbance of the redox system in T. japonicus. Among several oxidative stress-related genes, GSTs2b was significantly up-regulated in response to OA. These findings will be helpful for a better understanding on the potential impact of OA on life parameters and antioxidant system in the marine copepod T. japonicus. [ABSTRACT FROM AUTHOR]

Parabens are used as a preservative substance in a wide range of man-made products causing deleterious effects on aquatic organisms and therefore, the concern of their effects to aquatic organisms has been increased. In this study, acute toxicity of methylparaben (MeP), ethylparaben (EtP), and propylparaben (PrP) was assessed in the marine copepod Tigriopus japonicus. The acute toxicity assessment resulted in the median lethal concentration (LC50) values of MeP, EtP, and PrP were 29,754, 11,659, and 113 μg/L, respectively, for male and 38,183, 15,371, and 357 μg/L, respectively, for female, indicating the strongest toxicity of PrP, compared to MeP and EtP and the higher sensitivity of males compared to females. Developmental retardation and reproduction rate were also measured under chronic exposure. Furthermore, significant alteration in sex ratio was shown in PrP-exposed group, indicating PrP would have feminization effect in T. japonicus. Here we report different toxicity of three types of parabens and also shows potential estrogenic effects of PrP in T. japonicus. • LC50 values of MeP, EtP, and PrP were 29754, 11659, and 113 μg/L for male. • LC50 values of MeP, EtP, and PrP were 38183, 15371, and 357 μg/L for female. • The strongest toxicity was shown in PrP-exposed groups, compared to MeP and EtP. • Males showed the higher sensitivity compared to females. • Significant alteration in sex ratio was shown in PrP-exposed group in T. japonicus. [ABSTRACT FROM AUTHOR]

Highlights • In vivo toxicity of field-collected seawater on the rotifer Brachionus koreanus and copepod Tigriopus japonicus was assessed. • Metal-induced oxidative stress was assessed by analyzing the transcription and activities of antioxidant enzymes. • Several antioxidant genes were identified as useful biomarkers to detect early signs of metal pollution. Abstract Although many efforts have been made to understand the toxic effects of metals in aquatic invertebrates, there are limited data regarding metal toxicity in natural ecosystems, as most previous studies were conducted under controlled laboratory conditions. To address this data gap, we analyzed toxic effects and molecular responses in the marine rotifer Brachionus koreanus and the marine copepod Tigriopus japonicus following in vivo exposure to a seawater sample collected from a polluted region in South Korea. Inductively coupled plasma-mass spectrometry (ICP-MS) analysis of the field seawater sample found a variety of metals. Exposure to several dilutions of the field seawater sample impacted several endpoints in both species, including mortality and reproduction. Interestingly, the rotifer and copepod test species exhibited different patterns of effects on reactive oxygen species (ROS) and antioxidant enzymatic activities, suggesting that different regulatory mechanisms may be activated in the two species in response to exposure to toxic chemicals. Our study helps to better understand the defense mechanisms activated in aquatic invertebrates in response to metal-induced oxidative stress induced by contaminated seawater. [ABSTRACT FROM AUTHOR]

Abstract To find the adverse effects induced by cypermethrin, the ecotoxicological model copepods Tigriopus japonicus and Paracyclopina nana were exposed under cypermethrin, which is a widely used type-II pyrethroid insecticide in agriculture. Despite its large-scale application as insecticide in agriculture, little information is available on its adverse effects on aquatic invertebrates. In this study, the toxicity of cypermethrin on two copepods was assessed based on life parameters (growth rate and reproduction), oxidative stress and consequent antioxidant enzymatic activities, and gene expression profiles of antioxidants. The acute toxicity alone demonstrated that P. nana is less tolerant and sensitive against cypermethrin, compared to T. japonicus. However, under chronic exposure, life parameters of both P. nana and T. japonicus were severely affected by cypermethrin. Among antioxidant enzymatic activities, superoxide dismutase (SOD) and glutathione S -transferase (GST), in particular, demonstrated significant increase in response to cypermethrin. Furthermore, temporal-mRNA expression profile showed modulations in antioxidant related genes in response to cypermethrin. Our results provide the underlying toxic mechanism of cypermethrin and the oxidative stress defense mechanism and species-specific tolerance against cypermethrin in two model copepods species. Highlights • The toxicity of cypermethrin was assessed in vivo and in vitro endpoints. • T. japonicus was at a higher risk to cypermethrin compared to P. nana. • P. nana has well-developed detoxification system under chronic exposure. • The underlying toxic mechanism of cypermethrin was revealed in two copepods. • GST and SOD are likely useful biomarkers in response to cypermethrin. [ABSTRACT FROM AUTHOR]

Highlights • Body size-dependent tolerance to Cd toxicity was shown in three Brachionus spp. • Metabolic rate and Cd bioaccumulation support the body size-dependent tolerance in Brachionus spp. • MAPK signaling pathways and antioxidant enzymatic activities showed different activation patterns depending on the species. Abstract Although several studies have reported on different interspecific sensitivities in response to various toxicants, the response mechanisms are still poorly understood. Here, we investigate the interspecific toxicity of cadmium (Cd) and its mechanism using three marine rotifer Brachionus spp. that are distinguishable by body size, which is considered the most significant indicator of phenotypic difference. The body sizes of B. plicatilis , B. koreanus , and B. rotundiformis are significantly different throughout their life cycles (egg, neonate, and adult), with the smaller rotifer exhibiting higher sensitivity to Cd. To investigate the mechanisms that result in body size-dependent tolerance to Cd, metabolic and Cd bioaccumulation rates were investigated. Both rates have shown a significant correlation with body size, indicating that body size and its variables are important factors in determining Cd tolerance in Brachionus spp. In addition, similar patterns that further explain body size-dependent tolerance are shown in the phosphorylation status of mitogen-activated protein kinases, reactive oxygen species level, and antioxidant enzymatic activities. Our study provides valuable insight into size- and species-dependent toxicity mechanisms of species in the same genus. [ABSTRACT FROM AUTHOR]

Autophagy originated from the common ancestor of all life forms, and its function is highly conserved from yeast to humans. Autophagy plays a key role in various fundamental biological processes including defense, and has developed through serial interactions of multiple gene sets referred to as autophagy-related ( Atg ) genes. Despite their significance in metazoan life and evolution, few studies have been conducted to identify these genes in aquatic invertebrates. In this study, we identified whole Atg genes in four Brachionus rotifer spp., namely B. calyciflorus , B. koreanus , B. plicatilis , and B. rotundiformis , through searches of their entire genomes; and we annotated them according to the yeast nomenclature. Twenty-four genes orthologous to yeast genes were present in all of the Brachionus spp. while three additional gene duplicates were identified in the genome of B. koreanus , indicating that these genes had diversified during the speciation. Also, their transcriptional responses to cadmium exposure indicated regulation by cadmium-induced oxidative-stress–related signaling pathways. This study provides valuable information on 99 conserved Atg genes involved in autophagosome formation in Brachionus spp., with transcriptional modulation in response to cadmium, in the context of the role of autophagy in the damage response. [ABSTRACT FROM AUTHOR]

P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP) are ATP-binding cassette (ABC) transporters that confer multixenobiotic resistance (MXR) via their efflux activity, which enables a variety of xenobiotics to be expelled from cells. MXR has been proposed as the first line of defense against xenobiotics. In this study, the protective roles of P-gp and MRP in the rotifer Brachionus koreanus were examined in response to four biocides (alachlor, chlorpyrifos, endosulfan, and molinate) using fluorescent substrates and inhibitors specific to P-gp and MRP. Exposure of rotifers to the four biocides resulted in increased P-gp and MRP activity. Moreover, the rotifers became more sensitive to the biocides with a reduced tendency in survival and slower population growth rates, when P-gp or MRP was inhibited. These findings suggest that P-gp and MRP are involved in the defense system in response to biocide exposure. Furthermore, the transcriptional levels of the genes encoding P-gp and MRP were examined to uncover the mechanism by which MXR is regulated. Our results demonstrate a crucial role of the MXR efflux system in the defense response to biocides, thereby providing a better understanding of rotifer defense mechanisms on the molecular level. [ABSTRACT FROM AUTHOR]

Autophagy is a ‘self-eating’ system that regulates the degradation of cellular components and is involved in various biological processes including survival and development. However, despite its crucial role in organisms, the regulatory mechanism of autophagy remains largely unclear, particularly in invertebrates. In this study, conserved autophagy in the rotifer Brachionus koreanus in response to cadmium (Cd) exposure was verified by measuring acidic vesicle organelles using acridine orange (AO) and neutral red (NR) staining, and by detecting LC3 I/II on Western blot and immunofluorescence. We also demonstrated activation of p38 mitogen-activated protein kinase (MAPK) in response to Cd-induced oxidative stress, leading to the induction of autophagy in B. koreanus . This was further verified by analysis of MAPK protein levels and immunofluorescence of LC3 I/II after treatment with reactive oxygen species (ROS) scavengers and inhibitors specific to MAPKs. We propose a p38 MAPK-mediated regulatory mechanism of autophagy in B. koreanus in response to Cd-induced oxidative stress. This study will contribute to a better understanding of autophagic processes in invertebrates and its modulation by environmental stressors. [ABSTRACT FROM AUTHOR]

Recently, accidental spills of heavy oil have caused adverse effects in marine organisms. Oil pollution can induce damages on development and reproduction, linking with detrimental effects on diverse molecular levels of genes and proteins in plankton and fish. However, most information was mainly focused on marine vertebrates and consequently, limited information was available in marine invertebrates. Furthermore, there is still a lack of knowledge bridging in vivo endpoints with the functional regulation of cytochrome P450 ( CYP ) genes in response to oil spill pollution in marine invertebrates. In this paper, adverse effects of oil spill pollution in marine invertebrates are summarized with the importance of CYP genes as a potential biomarker, applying for environmental monitoring to detect oil spill using marine copepods. [ABSTRACT FROM AUTHOR]

The mitogen-activated protein kinases (MAPKs) family is known to mediate various biological processes in response to diverse environmental pollutants. Although MAPKs are well characterized and studied in vertebrates, in invertebrates the cross-reactivities of MAPKs antibodies were not clearly known in response to environmental pollutants due to limited information of antibody epitopes with material resources for invertebrates. In this paper, we performed phylogenetic analysis of MAPKs genes in the marine rotifer Brachionus koreanus and the copepods Paracyclopina nana and Tigriopus japonicus . Also in rotifer and copepods, several studies of Western blot of MAPK signaling pathways were shown in response to environmental pollutants, including multi-walled carbon nanotubes (MWCNTs), water-accommodated fractions (WAFs) of crude oil, and microplastics. This paper will provide a better understanding of the underlying mechanistic scenario in terms of cross-reactivities of mammalian antibodies in rotifer and copepod. [ABSTRACT FROM AUTHOR]

Unmanaged disposal of wastewater produced by underwater hull cleaning equipment (WHCE) is suspected to induce toxic effects to marine organisms because wastewater contains several anti-fouling compounds. To investigate the effects of WHCE on marine copepod, we examined the toxicity on life parameters (e.g. mortality, development, and fecundity) and gene expression changes of Tigriopus japonicus as model organism. Significant mortality and developmental time changes were observed in response to wastewater. No significant differences in fecundity were observed. Transcriptional profiling with differentially expressed genes from WHCE exposed T. japonicus showed WHCE may induce genotoxicity associated genes and pathways. In addition, potentially neurotoxic effects were evident following exposure to WHCE. The findings suggest that wastewater released during hull cleaning should be managed to reduce physiological and molecular deleterious effects in marine organisms. • 41 % of metals on average were reduced in 10 μm filtered hull cleaning wastewater. • The wastewaters caused mortality and retarded development in the copepod. • Genotoxic and neurotoxic effects were induced from unfiltered wastewater exposure. [ABSTRACT FROM AUTHOR]

2,2′,4,4′-tetrabromodiphenylether (BDE-47) is known to have the potential to disrupt the thyroid endocrine system in fishes due to its structural similarity to the thyroid hormones triiodothyronine (T3) and thyroxine (T4). However, the effects of BDE-47 on thyroid function in fishes remain unclear. In this study, abnormal development (e.g. deformity, hemorrhaging) and an imbalance in thyroid hormone (TH) homeostasis was shown in the early developmental stages of the mangrove killifish Kryptolebias marmoratus in response to BDE-47 exposure. To examine the thyroid endocrinal effect of BDE-47 exposure in mangrove killifish K. marmoratus larvae, transcript levels of genes involved in TH homeostasis and hypothalamus-pituitary-thyroid (HPT) axis-related genes were measured. The expression of thyroid hormone metabolism-related genes (e.g. deiodinases, UGT1ab ) and HPT axis-related genes was up-regulated and there were significant changes in TH levels ( P < 0.05) in response to BDE-47 exposure. This study provides insights into the regulation of TH homeostasis at the transcriptional level and provides a better understanding on the potential impacts of BDE-47 on the thyroid endocrine system of fishes. [ABSTRACT FROM AUTHOR]

Methylmercury (MeHg) is a concerning environmental pollutant that bioaccumulates and biomagnifies in the aquatic food web. However, the effects of MeHg on marine zooplankton are poorly understood even though zooplankton are considered key mediators of the bioaccumulation and biomagnification of MeHg in high-trophic marine organisms. Here, the toxicity of MeHg in the benthic copepod Tigriopus japonicus was assessed, and its adverse effects on growth rate and reproduction were demonstrated. Antioxidant enzymatic activities were increased in the presence of MeHg, indicating that these enzymes play an important role in the defense response to MeHg, which is regulated by a complex mechanism. Subsequent activation of different patterns of mitogen-activated protein kinase (MAPK) pathways was demonstrated, providing a mechanistic approach to understand the signaling pathways involved in the effects of MeHg. Our results provide valuable information for understanding the toxicity of MeHg and the underlying defense mechanism in response to MeHg exposure in marine zooplankton. [ABSTRACT FROM AUTHOR]

The ATP-binding cassette ( ABC ) transporter family is one of the largest gene family in animals, and members of this family are known to be involved in various biological processes due to their ability to transport a wide range of substrates across membranes using ATP cleavage-derived energy. We identified 61 ABC transporters in the genome of the monogonont rotifer Brachionus koreanus , and classified these into eight distinct subfamilies (A–H) by phylogenetic analysis. ABC transporters in the rotifer B. koreanus are comprised of 11 ABCA genes, 19 ABCB genes, 14 ABCC genes, 3 ABCD genes, 1 ABCE gene, 3 ABCF genes, 8 ABCG genes, and 2 ABCH genes. Extensive gene duplication and loss events in synteny were observed in several subfamilies. In particular, massive gene duplications of P-glycoproteins ( P-gps ), multidrug resistance proteins ( MRPs ), and Bk-Abcg-like proteins were observed. The ability of these B. koreanus proteins to function as multixenobiotic resistance (MXR) ABC transporters was validated using specific fluorescence substrates/inhibitors. The ABC transporter superfamily members identified in this study will be useful in future toxicological studies, and will facilitate comparative studies of the evolution of the ABC transporter superfamily in invertebrates. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Saxitoxin exposure induced mortality and behavioral changes in Tigriopus japonicus. • Genes involved in neuron system-related pathways were deregulated. • NRF2-mediated oxidative stress response was predicted as the main toxicity pathway. [ABSTRACT FROM AUTHOR]

To examine the toxic effects of multi-walled carbon nanotubes (MWCNTs) in the marine environment, we first exposed the monogonont rotifer ( Brachionus koreanus ) to MWCNTs in the presence of copper. The acute toxicity of copper decreased significantly with a decrease in copper bioavailability resulting from MWCNT exposure. Furthermore, we examined the effects of MWCNT exposure on reproductive capacity, population growth rate, growth patterns, antioxidant systems, and mitogen-activated protein kinase (MAPK) activation. Reproductive capacity, population growth rate, and body growth rate were significantly suppressed in B. koreanus in response to 1.3–4 mg/L MWCNT exposure. Furthermore, MWCNTs induced the generation of reactive oxygen species (ROS) and decreased the antioxidant enzymatic activities of catalase (CAT) and glutathione reductase (GR). However, the enzymatic activity of glutathione S -transferase (GST) was up-regulated after a 24 h-exposure to 100 mg/L MWCNTs. Exposure to 100 mg/L MCWNTs induced extracellular signal-regulated kinase (ERK) activation in B. koreanus , suggesting that p-ERK may mediate the adverse effects of MWCNTs in B. koreanus via the MAPK signaling pathway. Our results provide insight into the mechanistic basis of the ecotoxicological effects of MWCNTs in the marine environment. [ABSTRACT FROM AUTHOR]

In copepods, no information has been reported on the structure or molecular characterization of the nitric oxide synthase ( NOS ) gene. In the intertidal copepod Tigriopus japonicus , we identified a NOS gene that is involved in immune responses of vertebrates and invertebrates. In silico analyses revealed that nitric oxide (NO) synthase domains, such as the oxygenase and reductase domains, are highly conserved in the T. japonicus NOS gene. The T. japonicus NOS gene was highly transcribed in the nauplii stages, implying that it plays a role in protecting the host during the early developmental stages. To examine the involvement of the T. japonicus NOS gene in the innate immune response, the copepods were exposed to lipopolysaccharide (LPS) and two Vibrio sp. After exposure to different concentrations of LPS and Vibrio sp., T. japonicus NOS transcription was significantly increased over time in a dose-dependent manner, and the NO/nitrite concentration increased as well. Taken together, our findings suggest that T. japonicus NOS transcription is induced in response to an immune challenge as part of the conserved innate immunity. [ABSTRACT FROM AUTHOR]

Background Currently used techniques for diagnosing ischemic injury, such as magnetic resonance imaging and computed tomography, are not easily accessible for basic research using small animals due to their high cost and low availability. New method We investigated the dynamic recovery of infarct regions in ischemia-injured brains using indocyanine green (ICG), which is inexpensive and readily available. This dye was used to visualize blood vessels and infarct area, and to measure blood flow after a photothrombotic ischemic operation (PIO). Mice were injected with ICG via the tail vein, and a time-series of fluorescence signal images was acquired before and after PIO. We then applied color codes to arteries and veins in the images and analyzed ICG intensity and dynamics. Results These time-series stacked images showed changes in pial vessel morphology after PIO. Further, a map of maximum fluorescence intensity showed an infarct in the dorsal cortical region. Changes in the blood flow index and mean transit time were also observed in the infarct region after PIO. Comparison with existing method(s) Our application of ICG imaging provided a range of information on PIO-induced infarcts in mice with relative ease and in a cost-effective manner. Conclusion Our results show that optical imaging using ICG combined with a time-series analysis of molecular dynamics can be a useful tool for the anatomical and physiological monitoring of cortical ischemia. [ABSTRACT FROM AUTHOR]

• Arsenic (As) is a toxic metalloid that is widely distributed in the environment. • Arsenic toxicity is highly diverse and complex depending on its chemical form. • As toxicity is mainly associated with oxidative stress mechanisms. • AQP, HXT, phosphate channels, and ABC are involved in As influx/efflux systems. • Freshwater and marine organisms have As-specific biotransformation systems. Arsenic is a toxic metalloid that is widely distributed in the environment due to its persistence and accumulative properties. The occurrence, distribution, and biological effects of arsenic in aquatic environments have been extensively studied. Acute and chronic toxicities to arsenic are associated with fatal effects at the individual and molecular levels. The toxicity of arsenic in aquatic organisms depends on its speciation and concentration. In aquatic environments, inorganic arsenic is the dominant form. While trivalent arsenicals have greater toxicity compared with pentavalent arsenicals, inorganic arsenic can assume a variety of forms through biotransformation in aquatic organisms. Biotransformation mechanisms and speciation of arsenic have been studied, but few reports have addressed the relationships among speciation, toxicity, and bioavailability in biological systems. This paper reviews the modes of action of arsenic along with its toxic effects and distribution in an attempt to improve our understanding of the mechanisms of arsenic toxicity in aquatic organisms. [ABSTRACT FROM AUTHOR]

• As toxicity increased by NP co-exposure whereas it was alleviated by MP. • The highest level of As bioaccumulation was detected in NP + As groups. • No significant difference was shown for As metabolites. • Combined effects of As with NP or MP differ due to their different bioavailability. Besides the adverse biological effects induced by microplastics (MPs), the effects associated with sorption of ambient pollutants on MPs are considered as an emerging environmental problem as MPs act as a mediator of pollutants. The present study examines the combined effects of nano(micro)plastics (NMPs) and arsenic (As) by exposing the marine rotifer Brachionus plicatilis to MP particles at the micro-scale (6 μm) and nano-scale (nanoplastics, NPs) (50 nm) along with As. In vivo toxicity, bioaccumulation, and biochemical reactions were used to examine the effects of combined exposure. The results of in vivo experiments showed that As toxicity increased with NP exposure, whereas toxicity was alleviated by MPs, indicating a different mode of action between NPs and MPs in combination with As. The highest level of As bioaccumulation was detected in NP + As groups, and followed by MP + As and As-only exposure groups, whereas no significant difference between groups was shown for As metabolites. In addition, the activity of several ATP-binding cassette proteins that confer multixenobiotic resistance, which is responsible for efflux of As, was activated by As but significantly inhibited by NP exposure, supporting the findings of in vivo experiments. Our results show that the effects of combining exposure to As with NP and MPs differ depending on particle size and provide an in-depth understanding of both environmental pollutants. [ABSTRACT FROM AUTHOR]

Plastic is regarded as a major environmental concern. In particular, nanoplastics and microplastics (NMPs) are attracting global attention due to their potential impact on aquatic organisms. Here, we examined the effects of NMPs (50 nm polystyrene microbead nanoplastics [NPs] and 45 µm microplastics [MPs]) on oxidative status and gut microbiota in the marine medaka Oryzias melastigma. The NP-exposed group exhibited stronger oxidative stress with higher activation levels of antioxidants compared to the MP-exposed group. However, the MP-exposed group demonstrated induction of intestinal damage (e.g. , increased mucus ratio) with further alterations of gut microbiota, compared to the NP-exposed group. In particular, MPs caused more significant alterations of microbiota composition at both phylum and genus levels. Thus, in this study we show distinct toxicity pathways of NPs and MPs, an oxidative stress-mediated pathway (e.g. , antioxidants) induced by NP exposure and dysbiosis of gut microbiota in association with immune dysfunction induced by MP exposure. Our results are helpful for expanding our knowledge about the impacts of NMPs as potentially harmful substances in the aquatic environment. ga1 • Size-dependent effects of NMPs were detected in the gut and liver of marine medaka. • Nanoplastics induced oxidative stress-mediated responses. • Microplastics altered microbiota composition resulting in dysbiosis. [ABSTRACT FROM AUTHOR]

To assemble the genome of the marine water flea Diaphanosoma celebensis , a sentinel model for marine environmental monitoring, we constructed a high-quality genome using PromethION and HiSeq 2500 platforms. The total length of the assembled genome was 100.08 Mb, with N50 = 2.56 Mb (benchmarking universal single-copy orthologs, 96.9%) and consisted of 179 scaffolds. A total of 15,427 genes were annotated, and orthologous gene clusters in D. celebensis were analyzed and compared with those of the cladocerans Daphnia magna and Daphnia pulex. In addition, phase I, II, and III detoxification gene families of cytochrome P450s, glutathione S -transferases, and ATP-binding cassette were fully identified and revealed lineage-specific gene loss and/or expansion, suggesting that the evolution of detoxification gene families likely modulates fitness and susceptibility in response to environmental stressors. The study improves our understanding of the detoxification-related gene system and should contribute to future studies of molecular ecotoxicology in cladoceran species and their responses to emerging pollutants. Unlabelled Image • The total length of the assembled genome was 100.08 Mb, with N50 = 2.56 Mb (BUSCO 96.9%). • A total of 15,427 genes were annotated in Diaphanosoma celebensis. • Cytochrome P450s, glutathione S -transferases, and ATP-binding cassette proteins genes were fully identified. [ABSTRACT FROM AUTHOR]

The monogonont rotifer Brachionus spp. have been widely used for ecotoxicological studies because of their advantages as one of the most suitable laboratory experimental species. In the present study, we obtained and assembled the whole genome sequence of the rotifer Brachionus rotundiformis , consisting of 13,612 annotated genes with 213 scaffolds and 58 Mb in total length. Focusing on ecotoxicological aspects, we conducted a comparative genome analysis on the gene families involved in detoxification, including four to six sulfotransferase gene families, seven uridine 5′-diphospho-glucuronosyltransferase gene families, and 58, 61, or 70 ATP-binding cassette genes in the genus Brachionus including Brachionus koreanus and Brachionus plicatilis. Our results suggest that these gene families have undergone a species- and/or lineage-specific evolution in response to the surrounding environmental pressure. Our genome resource for B. rotundiformis would be highly useful for future ecotoxicological studies and also provides a better understanding on the view of evolutionary mechanism of detoxification in the genus Brachionus spp. Unlabelled Image • The rotifer Brachionus rotundiformis genome consisted of 13,612 genes with 213 scaffolds and 58 Mb in total length. • In Brachionus spp., we found four to six ST gene families, seven UGT gene families, and 58, 61, or 70 ABC genes. • These gene families have undergone a species- and/or lineage-specific evolution. • The genome resource for Brachionus rotundiformis would be highly useful for future ecotoxicological studies. • This provides a better understanding on the view of evolutionary mechanism of detoxification in the genus Brachionus spp. [ABSTRACT FROM AUTHOR]

Gluatathione S -transferases (GSTs) play a major role in phase II detoxification pathway to defend organisms in response to oxidative stress induced by xenobiotics and toxicants. To reveal the role of the recombinant GST zeta protein from the freshwater rotifer Brachionus calyciflorus , we isolated the zeta class GST in the freshwater rotifer B. calyciflorus. The recombinant B. calyciflorus GST zeta protein was highly expressed in the transformed Escherichia coli using pET28a vector. To determine its characteristics, effects of pH and temperature on B. calyciflorus GST zeta with enzymatic kinetics were also studied. In addition, a disk diffusion assay, uncovering the ability of transformed GST zeta in Escherichia coli , revealed that E. coli -transformed GST zeta significantly protected the transformed E. coli cells in response to oxidative stress induced by H 2 O 2 and metals such as mercury and cadmium. These results suggest that B. calyciflorus GST zeta recombinant protein is likely playing an important role to defend in response to metal-induced oxidative stress, providing a better understanding on the possible antioxidant role of GST zeta class in B. calyciflorus. Unlabelled Image • The Bc - GST zeta protein was highly expressed in the transformed Escherichia coli using pET28a vector. • Effects of pH and temperature on Bc - GST zeta protein with enzymatic kinetics were studied. • E. coli -transformed GST zeta significantly protected the transformed E. coli cells in response to oxidative stress. • Bc - GST zeta protein is likely playing an important role to defend in response to metal-induced oxidative stress. [ABSTRACT FROM AUTHOR]

To produce albinism in the marine medaka Oryzias melastigma , we disrupted the solute carrier family 45 (SLC45a2) gene by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 with a single guide RNA (sgRNA). Selected sgRNAs were able to target a SLC45a2 gene as confirmed by genotyping and heteroduplex mobility assay (HMA). Of the survived embryos after injection, 54.2% and 60.0% embryos exhibited albinism phenotype by sgRNA1 and sgRNA2, respectively. Deep sequencing at the on-target sites showed different insertion and deletion (indel) mutation profiles near the DNA cleavage sites, indicating high efficacy of producing SLC45a2 knock-out mutants by this method. Moreover, HMA at the potential off-target sites revealed that off-target activity would be induced at a low rate, or not induced at all. This albino marine medaka will be a good model for marine molecular ecotoxicology in establishment of diverse in vivo endpoints, and the application of this efficient gene targeting method in the marine medaka would be useful tool for mechanistic approaches. [ABSTRACT FROM AUTHOR]

Highlights • Fecundity and lifespan were decreased significantly (P < 0.05) in B[α]P-exposed B. rotundiformis. • Total intracellular ROS level and GST activity were significantly increased (P < 0.05). • Transcriptional regulation of GST s and ABC transporters were significantly upregulated and downregulated (P < 0.05). • B[α]P-exposed B. rotundiformis showed the increased levels of the p-JNK and p-p38. Abstract To examine the adverse effects of the benzo[α]pyrene (B[α]P), the monogonont rotifer Brachionus rotundiformis was exposed to various concentration of B[α]P (0 [control], 1, 10, and 100 μg/L) and measured life cycle parameters (e.g., mortality, fecundity [cumulated number of offspring], and lifespan), reactive oxygen species (ROS), antioxidant enzymatic activity of glutathione S -transferase (GST). In addition, defense-related transcripts (e.g., glutathione S -transferases [GSTs], ATP binding cassette [ABCs] transporters) and Western blot analysis of mitogen-activated protein kinase (MAPK) signaling pathway were investigated in B[α]P-exposed rotifer. In this study, the total intracellular ROS level and GST activity were significantly increased (P < 0.05), while fecundity and lifespan were also significantly (P < 0.05) reduced in a concentration dependent manner in B[α]P-exposed B. rotundiformis. In addition, transcriptional regulation of GST s and ABC transporters were significantly upregulated and downregulated (P < 0.05), respectively, suggesting that B[α]P can induce oxidative stress leading to induction of antioxidant system and detoxification mechanism. In addition to detoxification-related genes, B[α]P-exposed B. rotundiformis showed the increased levels of the p-JNK and p-p38, suggesting that B[α]P can activate MAPK signaling pathway in B. rotundiformis. [ABSTRACT FROM AUTHOR]

To evaluate the adverse effects of MeHg on the rotifer Brachionus koreanus and the copepod Paracyclopina nana , we assessed the effects of MeHg toxicity on life parameters (e.g. growth retardation and fecundity), antioxidant systems, and mitogen-activated protein kinase (MAPK) signaling pathways at various concentrations (1 ng/L, 10 ng/L, 100 ng/L, 500 ng/L, and 1000 ng/L). MeHg exposure resulted in the growth retardation with the increased ROS levels but decreased glutathione (GSH) levels in a dose-dependent manner in both B. koreanus and P. nana . Antioxidant enzymatic activities (e.g. glutathione S -transferase [GST], glutathione reductase [GR], and glutathione peroxidase [GPx]) in B. koreanus showed more positive responses compared the control but in P. nana , those antioxidant enzymatic activities showed subtle changes due to different no observed effect concentration (NOEC) values among the two species. Expression of antioxidant genes (e.g. superoxide dismutase [ SOD ], GSTs, glutathione peroxidase [ GPx ], and catalase [ CAT ]) also demonstrated similar effects as shown in antioxidant enzymatic activities. In B. koreanus , the level of p-ERK was decreased in the presence of 1000 ng/L MeHg, while the levels of p-ERK and p-p38 in P. nana were reduced in the presence of 10 ng/L MeHg. However, p-JNK levels were not altered by MeHg in B. koreanus and P. nana , compared to the corresponding controls. In summary, life parameters (e.g. reduced fecundity and survival rate) were closely associated with effects on the antioxidant system in response to MeHg. These observations provide a better understanding on the adverse effects of MeHg on in vivo life parameters and molecular defense mechanisms in B. koreanus and P. nana . [ABSTRACT FROM AUTHOR]

The ATP-binding cassette (ABC) protein superfamily is known to play a fundamental role in biological processes and is highly conserved across animal taxa. The ABC proteins function as active transporters for multiple substrates across the cellular membrane by ATP hydrolysis. As this superfamily is derived from a common ancestor, ABC genes have evolved via lineage-specific duplications through the process of adaptation. In this review, we summarized information about the ABC gene families in aquatic invertebrates, considering their evolution and putative functions in defense mechanisms. Phylogenetic analysis was conducted to examine the evolutionary significance of ABC gene families in aquatic invertebrates. Particularly, a massive expansion of multixenobiotic resistance (MXR)-mediated efflux transporters was identified in the absence of the ABCG2 ( BCRP ) gene in Ecdysozoa and Platyzoa, suggesting that a loss of Abcg2 gene occurred sporadically in these species during divergence of Protostome to Lophotrochozoa. Furthermore, in aquatic invertebrates, the ecotoxicological significance of MXR is discussed while considering the role of MXR-mediated efflux transporters in response to various environmental pollutants. [ABSTRACT FROM AUTHOR]

The ATP-binding cassette (ABC) protein superfamily is one of the largest gene families and is highly conserved in all domains. The ABC proteins play roles in several biological processes, including multi-xenobiotic resistance (MXR), by functioning as transporters in the cellular membrane. They also mediate the cellular efflux of a wide range of substrates against concentration gradients. In this study, 37 ABC genes belonging to eight distinct subfamilies were identified in the marine copepod Paracyclopina nana and annotated based on a phylogenetic analysis. Also, the functions of P-glycoproteins (P-gp) and multidrug resistance-associated proteins (MRPs), conferring MXR, were verified using fluorescent substrates and specific inhibitors. The activities of MXR-mediated ABC proteins and their transcriptional level were examined in response to polyaromatic hydrocarbons (PAHs), main components of the water-accommodated fraction. This study increases the understanding of the protective role of MXR in response to PAHs over the comparative evolution of ABC gene families. [ABSTRACT FROM AUTHOR]

Engineered multi-walled carbon nanotubes (MWCNTs) have received widespread applications in a broad variety of commercial products due to low production cost. Despite their significant commercial applications, CNTs are being discharged to aquatic ecosystem, leading a threat to aquatic life. Thus, we investigated the adverse effect of CNTs on the marine copepod Paracyclopina nana . Additional to the study on the uptake of CNTs and acute toxicity, adverse effects on life parameters ( e.g . growth, fecundity, and size) were analyzed in response to various concentrations of CNTs. Also, as a measurement of cellular damage, oxidative stress-related markers were examined in a time-dependent manner. Moreover, activation of redox-sensitive mitogen-activated protein kinase (MAPK) signaling pathways along with the phosphorylation pattern of extracellular signal-regulated kinase (ERK), p38, and c-Jun-N-terminal kinases (JNK) were analyzed to obtain a better understanding of molecular mechanism of oxidative stress-induced toxicity in the copepod P. nana . As a result, significant inhibition on life parameters and evoked antioxidant systems were observed without ROS induction. In addition, CNTs activated MAPK signaling pathway via ERK, suggesting that phosphorylated ERK ( p -ERK)-mediated adverse effects are the primary cause of in vitro and in vivo endpoints in response to CNTs exposure. Moreover, ROS-independent activation of MAPK signaling pathway was observed. These findings will provide a better understanding of the mode of action of CNTs on the copepod P. nana at cellular and molecular level and insight on possible ecotoxicological implications in the marine environment. [ABSTRACT FROM AUTHOR]

Vitellogenin (Vtg) is the precursor of egg yolk proteins, and its expression has been used as a reliable biomarker for estrogenic contamination in the aquatic environment. To examine the biomarker potential of the self-fertilizing killifish Kryptolebias marmoratus Vtgs (Km-Vtgs), full genomic DNAs of Km-Vtgs-Aa , Km-Vtgs-Ab , and Km-Vtgs-C were cloned, sequenced, and characterized. Three Vtg genes in K. marmoratus are tandemly placed in a 550 kb section of the same chromosome. In silico analysis of promoter regions revealed that both the Km-Vtgs-Aa and Km-Vtgs-Ab genes had an estrogen response element (ERE), but the Km-Vtgs-C gene did not. However, all three Km-Vtgs genes had several ERE-half sites in their promoter regions. Phylogenetic analysis demonstrated that the three deduced amino acid residues were highly conserved with conventional Vtgs protein, forming distinctive clades within teleost Vtgs. Liver tissue showed the highest expression of Km-Vtg transcripts in all tested tissues (brain/pituitary, eye, gonad, intestine, skin, and muscle) in response to endocrine disrupting chemical (EDC)-exposed conditions. Km-Vtg transcripts were significantly increased in response to 17β-estradiol (E2), tamoxifen (TMX), 4-n-nonylphenol (NP), bisphenol A (BPA), and octylphenol (OP) over 24 hr exposure. The Km-Vtg-A gene was highly expressed compared to the control in response to NP and OP. EDC-induced modulatory patterns of Km-Vtg gene expression were different depending on tissue, gender, and isoforms. [ABSTRACT FROM AUTHOR]

Multi-walled carbon nanotubes (MWCNTs) are nanoparticles widely applicable in various industrial fields. However, despite the usefulness of MWCNTs in industry, their oxidative stress-induced toxicity, combined toxicity with metal, and mitogen-activated protein kinase (MAPK) activation have not been widely investigated in marine organisms. We used the intertidal copepod Tigriopus japonicus as a test organism to demonstrate the adverse effects induced by MWCNTs in aquatic test organisms. The dispersion of the MWCNTs in seawater was maintained over 48 h without aggregation. MWCNTs caused a decrease in acute copper toxicity compared to the copper-only group in response to 20 and 100 mg/L MWCNTs, but not in response to 4 mg/L MWCNT, indicating that MWCNT may suppress acute copper toxicity. Reactive oxygen species (ROS) and enzymatic activities of glutathione S -transferase (GST) and catalase were significantly down-regulated in response to 100 mg/L MWCNT exposure. Glutathione (GSH) and glutathione reductase (GR) activity did not change significantly, indicating that MWCNTs may cause failure of the antioxidant system in T. japonicus . However, MWCNT induced extracellular signal-regulated kinase (ERK) activation without p38 and c-jun NH2-terminal kinase (JNK) activation, suggesting that ERK activation plays a key role in cell signaling pathways downstream of CNT exposure. This suggests that this pathway can be used as a biomarker for CNT exposure in T. japonicus . This study provides a better understanding of the cellular-damage response to MWCNTs. [ABSTRACT FROM AUTHOR]

The CYP2 genes are the largest and most diverse cytochrome P450 (CYP) subfamily in vertebrates. We have identified nine co-localized CYP2 genes (∼55 kb) in a new cluster in the genome of the highly resilient ecotoxicological fish model Kryptolebias marmoratus . Molecular characterization, temporal and tissue-specific expression pattern, and response to xenobiotics of these genes were examined. The CYP2 gene clusters were characterized and designated CYP2N22 - 23 , CYP2AD12 , and CYP2P16-20 . Gene synteny analysis confirmed that the cluster in K. marmoratus is similar to that found in other teleost fishes, including zebrafish. A gene duplication event with diverged catalytic function was observed in CYP2AD12 . Moreover, a high level of divergence in expression was observed among the co-localized genes. Phylogeny of the cluster suggested an orthologous relationship with similar genes in zebrafish and Japanese medaka. Gene expression analysis showed that CYP2P19 and CYP2N20 were consecutively expressed throughout embryonic development, whereas CYP2P18 was expressed in all adult tissues, suggesting that members of each CYP2 gene family have different physiological roles even though they are located in the same cluster. Among endocrine-disrupting chemicals (EDCs), benzo[α]pyrene (B[α]P) induced expression of CYP2N23 , bisphenol A (BPA) induced CYP2P18 and CYP2P19 , and 4-octylphenol (OP) induced CYP2AD12 , but there was no significant response to 4-nonylphenol (NP), implying differential catalytic roles of the enzyme. In this paper, we identify and characterize a CYP2 gene cluster in the mangrove killifish K. marmoratus with differing catalytic roles toward EDCs. Our findings provide insights on the roles of nine co-localized CYP2 genes and their catalytic functions for better understanding of chemical-biological interactions in fish. [ABSTRACT FROM AUTHOR]

2,2′,4,4′-Tetrabromodiphenyl ether (BDE-47) is widely dispersed endocrine disrupting chemicals (EDCs) in the aquatic ecosystem. Due to its devastating effect on marine organisms and insufficient database on toxicology, we investigated the adverse effects of BDE-47 on life parameters and antioxidant defense system following the reactive oxygen species (ROS) production in the monogonont rotifer Brachionus koreanus . In B. koreanus , the reduction in life cycle, fecundity, and population growth were observed in response to BDE-47. 50 μg/L BDE-47 significantly reduced ( P < 0.05) life expectancy and net reproductive rate. In response to 10–50 μg/L BDE-47 exposure, the oxidative stress was elicited via the generation of ROS, while the antioxidant related enzymes (e.g. glutathione S -transferase [GST] and glutathione reductase [GR]) have demonstrated significant activity levels ( P < 0.05) to further alleviate the oxidative stress in a concentration dependent manner. Furthermore, transcript profiles of antioxidant function ( GST-A , -O , and - S1–S8 )-related genes have shown the significant increase over 24 h in response to BDE-47 (0, 10, 25, and 50 μg/L). As for MAPK signaling pathway analysis, up-regulation of their activities was observed at 25 μg/L BDE-47 but their activities have reduced at adult NOEC concentration of 50 μg/L. This study provides a better understanding of the effects of BDE-47 on life parameters, molecular defense system, and activation of MAPK signaling pathway against generated oxidants in the rotifer. [ABSTRACT FROM AUTHOR]

Brominated flame retardant, 2, 2′, 4, 4′-tetrabromodiphenyl ether (BDE-47), has received grave concerns as a persistent organic pollutant, which is toxic to marine organisms, and a suspected link to endocrine abnormalities. Despite the wide distribution in the marine ecosystem, very little is known about the toxic impairments on marine organisms, particularly on invertebrates. Thus, we examined the adverse effects of BDE-47 on life history trait (development), oxidative markers, fatty acid composition, and lipid accumulation in response to BDE-47-induced stress in the marine copepod Paracyclopina nana . Also, activation level of mitogen-activated protein kinase (MAPK) signaling pathways along with the gene expression profile of de novo lipogenesis (DNL) pathways were addressed. As a result, BDE-47 induced oxidative stress ( e.g . reactive oxygen species, ROS) mediated activation of extracellular signal-regulated kinase (ERK) and c-Jun-N-terminal kinase (JNK) signaling cascades in MAPK pathways. Activated MAPK pathways, in turn, induced signal molecules that bind to the transcription factors (TFs) responsible for lipogenesis to EcR , SREBP , ChREBP promoters. Also, the stress stimulated the conversion of saturated fatty acids (SFAs) to polyunsaturated fatty acids (PUFAs), a preparedness of the organism to adapt the observed stress, which could be correlated with the elongase and desaturase gene ( e.g . ELO3 , Δ5-DES , Δ9-DES ) expressions, and then extended to the delayed early post-embryonic development and increased accumulation of lipid droplets in P. nana . This study will provide a better understanding of how BDE-47 effects on marine invertebrates particularly on the copepods, an important link in the marine food chain. [ABSTRACT FROM AUTHOR]

De novo lipogenesis (DNL) is a fatty acid synthesis process that requires several genes, including sterol regulatory element binding protein (SREBP), ATP-citrate lyase (ACLY), and acetyl-CoA carboxylase (ACC). DNL up-regulation is able to induce fat accumulation through an increase in fatty acids. To investigate the relationship between DNL up-regulation and the accumulation of fatty acids and lipid droplets in response to 2,2′,4,4′ tetrabrominated diphenyl ether (BDE-47), we examined DNL in the copepod Tigriopus japonicus . Transcription levels of DNL-related genes were increased after exposure to 2.5 μg/L BDE-47 for 24 h. After exposure to 2.5 μg/L BDE-47, palmitic acid was significantly increased ( P < 0.05) at days 1 and 4, along with upregulation of fatty acid synthesis-related genes ( e.g. , desaturases and elongases). However, docosahexaenoic acid and arachidonic acid were down-regulated at days 1 and 4, showing an antagonistic effect. Lipid droplet area significantly increased in Nile red staining analysis after 24 h of exposure to 2.5 μg/L BDE-47 in T. japonicus , while DNL was down-regulated in response to 500 μM salicylate (a lipogenesis inhibitor), indicating that BDE-47 exposure is closely associated with an increase in fatty acids in this copepod. This study provides a better understanding of the effects of BDE-47 on DNL in copepods. [ABSTRACT FROM AUTHOR]

• We assembled the genome of the Daphnia magna NIES strain. • The draft genome of the Daphnia magna NIES strain was built into 10 LGs, comprising a genome size of 173.47 Mb. • The N50 value of the genome was 12.54 Mb, and the BUSCO value was 98.8%. • In the D. magna NIES genome, 15,684 genes were functionally annotated. • CRISPR/Cas9-mediated GST-O2 gene targeting has been made with an efficient make-up (25.0%) of mutant lines. The water flea Daphnia magna is a small freshwater planktonic animal in the Cladocera. In this study, we assembled the genome of the D. magna NIES strain, which is widely used for gene targeting but has no reported genome. We used the long-read sequenced data of the Oxford nanopore sequencing tool for assembly. Using 3,231 genetic markers, the draft genome of the D. magna NIES strain was built into ten linkage groups (LGs) with 483 unanchored contigs, comprising a genome size of 173.47 Mb. The N50 value of the genome was 12.54 Mb and the benchmarking universal single-copy ortholog value was 98.8%. Repeat elements in the D. magna NIES genome were 40.8%, which was larger than other Daphnia spp. In the D. magna NIES genome, 15,684 genes were functionally annotated. To assess the genome of the D. magna NIES strain for CRISPR/Cas9 gene targeting, we selected glutathione S -transferase omega 2 (GST-O2), which is an important gene for the biotransformation of arsenic in aquatic organisms, and targeted it with an efficient make-up (25.0%) of mutant lines. In addition, we measured reactive oxygen species and antioxidant enzymatic activity between wild type and a mutant of the GST-O2 targeted D. magna NIES strain in response to arsenic. In this study, we present the genome of the D. magna NIES strain using GST-O2 as an example of gene targeting, which will contribute to the construction of deletion mutants by CRISPR/Cas9 technology. [ABSTRACT FROM AUTHOR]

• No significant change in survival rate was observed in response to G. catenatum. • Swimming speed and swimming distance of Artemia franciscana were significantly decreased in response to G. catenatum. • No significant change in swimming path trajectory of brine shrimp was observed in response to G. catenatum. • A. franciscana-hsp genes were upregulated in response to 600 cells/mL G. catenatum. To understand the effects of the toxic marine dinoflagellate, Gymnodinium catenatum, on the brine shrimp, Artemia franciscana , we examined the acute toxicity and swimming behavior parameters such as swimming speed, swimming distance, and swimming path trajectory with transcriptional regulation of heat shock protein (hsp) genes in response to G. catenatum exposure. Mortality was not observed in response to G. catenatum. In the case of swimming behavior parameters, swimming speed and swimming distance were significantly decreased (P < 0.05) for 5 min at three concentrations (240, 360, and 600 cells/mL) of G. catenatum , whereas no significant change in swimming path trajectory was observed, suggesting that G. catenatum has potential adverse effects on the swimming behavior of A. franciscana. Additionally, the four A. franciscana - hsp genes (hsp26, hsp40, hsp70 , and hsp90) were upregulated in response to G. catenatum. In particular, A. franciscana - hsp40 was significantly upregulated in response to 600 cells/mL G. catenatum , suggesting that A. franciscana - hsp genes are highly associated with cellular defense mechanisms and that A. franciscana - hsp40 is a potential biomarker for G. catenatum exposure. Overall, this study improves our understanding of the effects of G. catenatum on the swimming behavior and cellular defense mechanisms of A. franciscana. [ABSTRACT FROM AUTHOR]

Metal pollution is one of the major threats to the aquatic environment due to its high bio-concentrations and toxicity. Although numerous studies have been conducted to understand detoxification and toxicity mechanisms in aquatic invertebrates, most of them were limited in laboratory study. Here, we investigated adverse effects of the contaminated-natural seawater on the marine rotifer Brachionus koreanus. Field seawater was collected from five different sites of Youngil Bay, South Korea where pollution by metals is predicted due to industrial discharges from nearby steel industry complexes. The marine rotifer B. koreanus was exposed to different seawater samples, and we found decreased population growth rates with highly induced transcriptional level of detoxification-related genes. Our finding shows a link between the induction of metal-mediated regulation of detoxification genes in B. koreanus and concentration of heavy metals present in various seawater samples, implying those biomarker genes can be useful to assess the toxicity of metal polluted natural seawaters. Our results are helpful to validate and establish biomarker genes for early risk assessment of metal contamination in the natural sea water. Unlabelled Image • ICP-MS analysis revealed the presence of nickel, copper, zinc, and lead in Youngil Bay, South Korea. • Decreased population growth rates were shown in B. koreanus upon seawater exposures. • Detoxification-related genes were highly induced upon seawater exposures. • The induction of detoxification genes was correlated with concentration of heavy metals in various seawaters. [ABSTRACT FROM AUTHOR]