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Microplastics have become a worldwide pollutant, widely discovered in soil, air and aquatic environment. Microplastics have been found in habitats where crayfish (Procambarus clarkii) cultivated, but the impact of microplastics on crayfish remains unclear. In this study, after 21-day dietary exposure, polyethylene (PE) particles were found to accumulate in intestine, hepatopancreas, gills and hemolymph of crayfish. Furthermore, PE particles can still be detected in these tissues after a 7-day depuration in clean water. PE retained in these tissues caused oxidative stress responses, as indicated by the change of oxidative-stress-related index, such as the increase of H 2 O 2 level and SOD activity. PE exposure also caused hemocytic encapsulation in crayfish hepatopancreas and increase of mucus secretion in intestine. Moreover, PE exposure affected the microbiota balance in crayfish, by reducing the total microbiota abundance and altering the proportions of many bacterial families. Interestingly, results showed that PE exposure led to of lower numbers of hemocytes and declination of phenoloxidase activity. Finally, PE exposure induced the expression of immune-related genes, including transcription factors and antimicrobial peptides. Taken these together, we conclude that PE microplastics exert considerable toxic effects on crayfish and are a potential threat to crayfish aquaculture and consumption. This study provides basic toxicological data toward quantifying and illuminating the impact of PE microplastics on freshwater animals. [Display omitted] • Polystyrene microplastics accumulated in several tissues of crayfish and caused histopathological changes. • Polystyrene microplastics induced oxidative stress and immune responses in crayfish. • Polystyrene microplastics induced gut microbiota dysbiosis in crayfish. [ABSTRACT FROM AUTHOR]

Due to sexual dimorphism in growth traits, all-female populations and populations with a high proportion of females hold significant economic value in common carp (Cyprinus carpio L.). However, the underlying mechanisms governing gonadal differentiation remain inadequately understood. NF-κB, as a crucial regulator of anti-apoptotic processes, plays a significant role in mammalian gonad formation and zebrafish oocyte maintenance. To investigate the potential roles of the NF-κB pathway in the gonad differentiation of common carp, juveniles with undifferentiated gonads were treated with an activator (DOC) or inhibitor (NAI) of NF-κB. Our findings revealed that juveniles exposed to DOC exhibited a significantly increased female-to-male ratio (from 0.97:1 to 4.30:1) and substantial increases in weight and body length. Conversely, inhibiting NF-κB with NAI resulted in a decreased female-to-male ratio (from 1.06:1 to 0.33:1). Furthermore, NAI also induced the occurrence of ovotestis. Additional in vivo and ex vivo experiments demonstrated that DOC activation of the NF-κB pathways led to inhibition of apoptosis, significant increases in serum 17β-estradiol (E2) levels and the E2/T ratio, and upregulation of the female-biased genes foxl2 and cyp19a1a. Simultaneously, it significantly suppressed serum 11-ketotestosterone (11-KT) levels and the expression of male-biased genes amh , dmrt1 , and gsdf and induced VTG levels in the hepatopancreas of male carp. However, NAI exhibited contrasting effects. Both DOC and NAI disrupted the balance of the hypothalamus-pituitary-gonad (HPG) reproductive axis at multiple potential loci, including changes in the expression levels of steroid hormone synthesis-related genes in gonads. Transcriptomic analysis revealed comprehensive molecular changes in undifferentiated gonads during DOC-induced feminization and NAI-induced masculinization. The transcriptome results were consistent with the observed alterations in hormone content and the expression of genes involved in sex hormone synthesis, inflammation, and apoptosis following exposure to DOC or NAI. These findings suggest the involvement of NF-κB in the sex differentiation of common carp by regulating apoptosis and disrupting the HPG reproductive axis. Our study also explores a DOC-induced method of producing high ratios of females and discovers the underlying mechanism of NF-κB's contribution to sex differentiation in common carp. • DOC induces high ratios of females and promotes the growth of common carp. • NAI results in a male-biased sex ratio and the appearance of ovotestes. • Apoptosis is involved in the process of feminization caused by activation of NF-κB. • The imbalance of the HPG axis contributes to the skewed sex ratio. [ABSTRACT FROM AUTHOR]

The levels of the chlorinated organic compound Dechlorane Plus (DP) are increasing in aquatic ecosystems. To investigate the adverse effects of DP on aquatic animals, common carp (Cyprinus carpio) were subjected to three different DP concentrations (30 μg L−1, 60 μg L−1, and 120 μg L−1) for 1 d, 15 d, and 30 d. Histology and the hepatic and cerebral expression levels of several key antioxidant, detoxification, and apoptotic factors were then examined. Histopathological inspections showed that the liver and brain were severely damaged in carp exposed to 60 μg L−1 and 120 μg L−1 DP. Relative to the controls, the superoxide dismutase and glutathione activity levels and the malondialdehyde content were also changed in livers and brains exposed to DP. Besides, significant alterations in the expression levels of the inflammatory cytokines IL-1β, IL-6, and IL-10 were observed in the livers of carp subjected to DP. Relative to the control, the brains of DP-exposed carp presented with significantly upregulated IL-1β and IL-6 in carp treated with 120 μg L−1 DP for 30 d. The transcription levels of hepatic cyp2b4, cyp1b1, and cyp3a138 were all increased compared with the untreated at all DP exposure concentrations. The aforementioned results suggest that DP exposure perturbs fish metabolism and causes liver injury by inhibiting antioxidant enzyme activity, increasing lipid peroxidation, promoting inflammation, and inducing cell apoptosis. This information and the analytical methodology used to acquire it may form the basis for future ecological risk assessments on DP and related xenobiotics in aquatic animals. [ABSTRACT FROM AUTHOR]

Nanomaterials have attracted much attention in the biomedical field. Black phosphorus quantum dots (BPQDs) have shown great potential in biomedical applications, but their potential risks to biosafety and environmental stability have not been fully evaluated. In the present study, zebrafish (Danio rerio) embryos were exposed to 0, 2.5, 5 and 10 mg/L BPQDs from 2 to 144 h post-fertilization (hpf) to explore developmental toxicity. The results showed that exposure to BPQDs for 96 h induced developmental malformations (tail deformation, yolk sac edema, pericardial edema, and spinal curvature) in zebrafish embryos. ROS and antioxidant enzyme activities (CAT, SOD, MDA and T-AOC) were substantially altered and the acetylcholinesterase (AChE) enzyme activity was significantly decreased in the BPQDs exposed groups. Locomotor behavior was inhibited after BPQDs exposure for 144 h in zebrafish larvae. A significant increase in 8-OHdG content indicates DNA oxidative damage in embryos. In addition, obvious apoptotic fluorescence signals were detected in the brain, spine, yolk sac and heart. At the molecular level, the mRNA transcript levels of key genes related to skeletal development (igf1, gh, MyoD and LOX), neurodevelopment (gfap, pomca, bdnf and Mbpa), cardiovascular development (Myh6, Nkx2.5, Myl7, Tbx2b, Tbx5 and Gata4) and apoptosis (p53, Bax, Bcl-2, apaf1, caspase-3 and caspase-9) were abnormal after BPQDs exposure. In conclusion, BPQDs induced morphological malformations, oxidative stress, locomotor behavior disorders, DNA oxidative damage and apoptosis in zebrafish embryos. This study provides a basis for further study on the toxic effects of BPQDs. [Display omitted] • BPQDs caused developmental toxicity in the zebrafish embryos. • BPQDs induced oxidative stress, oxidative DNA damage and apoptosis in zebrafish embryos. • BPQDs induced developmental neurotoxicity and decreased locomotor behavior. [ABSTRACT FROM AUTHOR]

Abstract Ionic liquids have garnered increasing attention due to their capacity for low vapor pressure, lack of flammability, designability, good stability, and as a asubstitute for conventional organic solvents. However, their toxicity to various organisms has caused growing concern in recent years. Our study aims to evaluate the toxicity of 1-hexadecyl-3-methylimidazolium chloride ([C 16 min]Cl) to human hepatocellular carcinoma (HepG2) cells, including cell viability, genotoxicity, oxidative stress, apoptosis, cell cycle, and apoptosis-related gene expression. Our results with HepG2 cells suggested that [C 16 min]Cl inhibited cellular growth, decreased cell viability, induced DNA damage and apoptosis, inhibited superoxide dismutase, decreased glutathione content, increased cellular malondialdehyde levels as well as altering the cell cycle. Moreover, the induction of [C 16 min]Cl altered the transcription of p53 , Bax and Bcl-2 , which are critical for controlling cell cycles progression and death, which suggests its involvement with cytotoxicity and apoptosis induced by [C 16 min]Cl in HepG2 cells. Taken together, these results revealed that [C 16 min]Cl exerted genotoxicity, oxidative stress and induced apoptosis in HepG2 cells; hence, it is not a healthy solvent. Highlights • [C 16 min]Cl is cytotoxic to HepG2 cells. • [C 16 min]Cl inhibits HepG2 cells growth and decreases their viabilities. • [C 16 min]Cl exerts genotoxicity, oxidative stress and induces apoptosis in HepG2 cells. [ABSTRACT FROM AUTHOR]

Sox4, a member of the SoxC subfamily which of the Sox family, plays important roles in the development of the vertebrate gonad and nervous system. We have cloned a Sox4 homologue from the brain of Misgurnus anguillicaudatus using homologous cloning and rapid amplification of cDNA ends. We named the cloned gene as MaSox4. The full-length cDNA was 2122 bp, containing a 718 bp 5′-untranslated region and a 267 bp 3′-untranslated region. The open-reading frame of the cloned gene encoded 378 amino acids and contained a characteristic HMG-box DNA-binding domain with the specific motif (RPMNAFMVW). Phylogenetic analysis indicated that MaSox4 is highly homologous to Sox4 in different species. Protein sequence analysis showed that MaSox4 is a nonsecretory hydrophilic protein. Quantitative real-time reverse transcription polymerase chain reaction and in situ hybridization assay revealed that MaSox4 was ubiquitously expressed during embryogenesis and is present in various adult tissues, especially in the central nervous system. Our study suggests that MaSox4 is highly conserved among vertebrates’ evolution and might be involved in developmental processes such as embryogenesis, neurogenesis and gonad development. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) are members of humoral immunity and particpate in resisting microbial invasion. In this study, an AMP gene hepcidin was obtained from the oriental loach Misgurnus anguillicaudatus and named Ma-Hep. This Ma-Hep encodes a peptide of 90 amino acids, with a predicted active peptide segment (Ma-sHep) of 25 amino acids at C terminus. Stimulation by a bacterial pathogen Aeromonas hydrophila resulted in significant up-regulation of Ma-Hep transcripts in loach midgut, head kidney, and gill. Ma-Hep and Ma-sHep proteins were expressed in Pichia pastoris and their antibacterial activity was examined. Results showed that Ma-sHep possessed stronger antibacterial activity against various Gram-positive and Gram-negative bacteria, compared to Ma-Hep. Scanning electron microscopy showed that Ma-sHep might kill bacteria by destroying bacterial cell membranes. Moreover, we found that Ma-sHep had an inhibitory effect on blood cell apoptosis induced by A. hydrophila and facilitated the bacterial phagocytosis and clearance in loach. Histopathological analysis indicated Ma-sHep could protect liver and gut of loach from bacterial infection. Ma-sHep has high thermal stability and PH stability, which is conducive to further feed addition. Feed supplemented with Ma-sHep expressing yeast improved the intestinal flora of loach by increasing the dominant bacteria and decreasing the harmful bacteria. Feed supplemented with Ma-sHep expressing yeast also regulated the expression of inflammatory related factors in various tissues of loach and reduced the mortality of loach upon bacterial infection. These findings show that the antibacterial peptide Ma-sHep is involved in the antibacterial defense of loach and can be used as a candidate for new antimicrobial agents in aquaculture. • The Hepcidin from Misgurnus anguillicaudatus was cloned and identified. • Ma-sHep possessed strong antibacterial activity against various bacteria. • Ma-sHep had an inhibitory effect on blood cell apoptosis induced by A. hydrophila. • Ma-sHep could protect loach tissues from bacterial infection. • Feed supplemented with Ma-sHep expressing yeast enhance immune capacity of loach. [ABSTRACT FROM AUTHOR]

• The gonadal development was remarkably hampered by PVC microplastics. • PVC microplastics damaged histological structure of gonads and brains. • Disruption effects of PVC microplastics were observed in sex hormone levels. • Genes transcription and translation in HPG axis were changed by PVC microplastics The negative consequences of microplastics pollution on the health of aquatic species have garnered extensive attention. However, the mechanisms through which microplastics may cause harm in the reproductive processes of fish remain unknown. For this study, Cyprinus carpio var. was subjected to four treatments with various concentrations of PVC microplastics for 60 days, through food rationed diets (no plastic control, 10%, 20% and 30%). The gonadosomatic indices, gonad and brain histologies, sex hormone levels, and transcriptional and translational genes in the hypothalamic-pituitary-gonadal (HPG) axes of both sexes were observed. According to the results, the gonadosomatic indices were significantly decreased, gonadal development was delayed, and the level of estradiol (E2) in the females was significantly elevated. In addition, the expression levels of genes associated with the HPG axis in the brains and gonads (gnrh, gtha1, fshβ, cyp19b, erα, vtg1, dmrt1, sox9b, and cyp19a) and the transcription levels of apoptosis-related genes in the brains and gonads (caspase3, bax , and bcl-2) exhibited significant changes. Further investigation revealed that the translation levels of genes linked to sex differentiation and sex steroid hormone (cyp19b and dmrt1) were significantly altered. These findings indicated that PVC likely microplastics may have a negative impact on the reproductive system of Cyprinus carpio var. by inhibiting gonadal development, affecting the gonad and brain structures, and altering the levels of steroid hormones and the expression of HPG axis-related genes. This work provides new insights into the toxicity of microplastics in aquatic organisms by revealing that PVC microplastics are a potential threat against the reproduction of fish populations. [ABSTRACT FROM AUTHOR]

A number of studies have established that in vertebrates, Sox3 is involved in a wide range of developmental processes, including sex differentiation and neurogenesis. However, the exact functions of the Sox3 gene have not been documented so far in teleosts. Here, we cloned the full length cDNA of Sox3 from the teleost fish, Paramisgurnus dabryanus, which we designated PdSox3. Sequence analysis revealed that PdSox3 encodes a hydrophilic protein, and shares high homology with Sox3 in other species, ranging from mammals to fishes. Quantitative real‐time reverse transcription PCR, and in situ hybridization showed that PdSox3 is consistently expressed during embryogenesis, mainly localized in the developing central nervous system. Tissue distribution analyses revealed that PdSox3 is abundant in the adult brain, especially in particle cell layer. Furthermore, PdSox3 expression was higher in gonads, in primary spermatocyte cells, primary oocytes, and previtellogenic oocyte cells. All of these results suggest that PdSox3 plays an important role in early embryonic development, in particular the formation and development of the nervous system, and gonad development, similarly to other vertebrates. This is the first report describing Sox3 gene expression from this species, and the results are necessary to provide fundamental information on both the functional and evolutionary role of Sox3 across different species. [ABSTRACT FROM AUTHOR]

Sox7, Sox17 and Sox18 are the members of the Sry-related high-mobility group box family (SoxF) of transcription factors. SoxF factors regulate endothelial cell fate as well as development and differentiation of blood cells and lymphatic vessels. There is very less information about the functions of these genes in fish. We obtained the full-length cDNA sequence of SoxF genes including Sox7, Sox17 and Sox18 in Cyprinus carpio, where Sox7 and Sox18 had two copies. The construction of a phylogenetic tree showed that these genes were homologous to the genes in other species. Chromosome synteny analysis indicated that the gene order of Sox7 and Sox18 was highly conserved in fish. However, immense change in genomic sequences around Sox17 had taken place. Numerous putative transcription factor binding sites were identified in the 5′ flanking regions of SoxF genes which may be involved in the regulation of the nervous system, vascular epidermal differentiation and embryonic development. The expression levels of SoxF genes were highest in gastrula, and was abundantly expressed in the adult brain. We investigated the expression levels of SoxF genes in five specific parts of the brain. The expression levels of Sox7 and Sox18 were highest in the mesencephalon, while the expression level of Sox17 was highest in the epencephalon. In carp, the expression patterns of SoxF genes indicated a potential function of these genes in neurogenesis and in vascular development. These results provide new information for further studies on the potential functions of SoxF genes in carp. [ABSTRACT FROM AUTHOR]

Ovary development is a complex process involving numerous genes; the molecular mechanism underlying the ovary development of carp is still unknown. Here we used Illumina HiSeq™ 2500 to explore the transcriptome of undifferentiated gland (PG), juvenile ovary (OJ) and adult ovary (OA) of Yellow River carp ( Cyprinus carpio ). A total of 58,749 unigenes were obtained, comprising 45,707 known genes and 13,042 new genes. We identified differentially-expressed genes (DEGs) during development and characterized the functional properties of DEGs by comparison with the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes databases. qRT-PCR was used to analyze the expression of 22 DEGs and the results corresponded with those of RNA-Seq. Among DEGs between PG and OJ, some upstream regulators of gonad development were upregulated in PG, such as cyp19a and sox9 , while some oocyte-specific genes were upregulated in OJ, such as nobox, bmp15 and zp2 . Among DEGs between OJ and OA, many oocyte physiological function-related genes were upregulated in OA, such as fem-1 and foxl2 . GO analysis showed a higher number of DEGs from PG–OJ analysis were assigned to reproduction terms. Furthermore, our investigation has also revealed DEGs identified from PG–OJ analysis were enriched in several important functional pathways, such as Fanconi anemia and the notch signal pathway. These data suggested a dynamic shift in gene expression during ovary development, and DEGs between PG and OJ provided crucial candidate gene data for the study of ovarian differentiation. Additionally, a total of 1,776,769 single nucleotide polymorphisms and 157,279 INDEs were revealed from transcriptome data. This result will contribute to knowledge of ovary differentiation of Yellow River carp. [ABSTRACT FROM AUTHOR]

Sox4 belonged to the SoxC subfamily of the Sox family, which play important roles in the development of the vertebrate gonad and nervous system. A Sox4 homologue was cloned from brain of Paramisgurnus dabryanus by using homologous cloning and rapid amplification of cDNA ends (RACE), designated as PdSox4 . The full-length cDNA was 2163bp, containing the 759bp 5'-untranslated region, 267bp 3'-untranslated region and encoding a putative protein of 378 amino acids with a characteristic high mobility group box (HMG-box) DNA-binding domain of 79 amino acids with the specific motif (RPMNAFMVW). Alignment and phylogenetic analyses indicated that PdSox4 shares highly identical sequence with Sox4 homologues from different species. The signal peptide analysis predicted that PdSox4 is a non-secretory protein. The hydropathy profile of PdSox4 protein revealed that this protein is hydrophilic in nature. The expression profiles of PdSox4 in different developmental stages and various adult tissues of sexs were analyzed by quantitative real-time RT-PCR (qRT-PCR) and In situ hybridization (ISH). The results showed that PdSox4 was ubiquitously expressed during embryogenesis and various adult tissues, especially in central nervous system. Tissue distribution analyses revealed that PdSox4 was expression in developing germ cells. Taken together, these preliminary findings suggested that PdSox4 is highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, neurogenesis and gonad development. [ABSTRACT FROM AUTHOR]

Teleost fins can regenerate accurate position-matched structure and function after amputation. However, we still lack systematic transcriptional profiling and methodologies to understand the molecular basis of fin regeneration. After histological analysis, we established a suppression subtraction hybridization library containing 418 distinct sequences expressed differentially during the process of blastema formation and differentiation in caudal fin regeneration. Genome ontology and comparative analysis of differential distribution of our data and the reference zebrafish genome showed notable subcategories, including multi-organism processes, response to stimuli, extracellular matrix, antioxidant activity, and cell junction function. KEGG pathway analysis allowed the effective identification of relevant genes in those pathways involved in tissue morphogenesis and regeneration, including tight junction, cell adhesion molecules, mTOR and Jak-STAT signaling pathway. From relevant function subcategories and signaling pathways, 78 clones were examined for further Southern-blot hybridization. Then, 17 genes were chosen and characterized using semi-quantitative PCR. Then 4 candidate genes were identified, including F11r , Mmp9 , Agr2 and one without a match to any database. After real-time quantitative PCR, the results showed obvious expression changes in different periods of caudal fin regeneration. We can assume that the 4 candidates, likely valuable genes associated with fin regeneration, deserve additional attention. Thus, our study demonstrated how to investigate the transcript profiles with an emphasis on bioinformatics intervention and how to identify potential genes related to fin regeneration processes. The results also provide a foundation or knowledge for further research into genes and molecular mechanisms of fin regeneration. [ABSTRACT FROM AUTHOR]

Pollution of microplastics (MPs) has become a global environmental issue due to the difficulty in its degradation and may cause unexpected ecological effects. Nevertheless, little is known about the potential effects of MPs on reproduction toxicity in aquatic species. In this study, adult loach (Paramisgurnus dabryanus, F0 generation) were exposed to two concentrations (1 and 10 mg/L) of polyethylene MPs (PE-MPs) for 15 or 30 days, and the toxic effects in parental loach and the offspring (F1 generation) were examined. Our results showed that PE-MPs exposure could change the indicators content of antioxidant system in the brain, liver, and gonad. PE-MPs can accumulate in the gonads, disrupt the transcription of HPG-axis related genes, alter sex hormone levels, increase cell apoptosis and gonadal pathological lesions, lead to the damage of biological characteristics of semen, and affect the reproduction in F0 generation. PE-MPs remaining in the parental gonads can be transferred to the F1 generation embryos and accumulated on the embryonic chorionic membrane, increasing mortality and malformation rates, accelerating hatching time, and decreasing hatching rate and body length. These results suggest that PE-MPs leads to a potential adverse influence on reproduction and serious impacts on population sustainability. This work provides a new perspective into the effects of MPs on reproductive damage and cross-generational effects in teleost fish, which have implications in fields of freshwater ecology and environmental toxicology. [Display omitted] • PE-MPs could remain in the gonads and destroyed the reproductive organs. • PE-MPs could disturb the antioxidant, immune and endocrine system. • PE-MPs caused germ cell apoptosis and gamete quality decline. • PE-MPs transferred to the embryo chorionic membrane by parental transmission. • Parental exposure resulted in developmental toxicity of F1 larvae. [ABSTRACT FROM AUTHOR]

Hsp70 is the most strongly induced in response to various cellular stresses and a good candidate for investigating its role in tissue injury. We firstly cloned full-length cDNA of hsp70 from Paramisgurnus dabryanus ( PdHsp70 ) by RACE method (GenBank: KP402408.1 ). Then regeneration and inflammation of fin were established by amputation and scratch respectively. Quantitative RT-PCR detected the PdHsp70 began to increase rapidly its expression at 1 days post amputation (dpa) and reached the peak at 2 dpa during fin regeneration. Its expression was also up-regulated at 2 days post scratch (dps) of inflammation but still significant weaker in comparison with it in regenerated fin at 2 dpa. Next, immunohistochemistry analysis of PdHsp70 showed that PdHsp70 located mainly in the deeper epidermis of regenerated fin and was stronger than its expression in the scratched inflammatory fin which was involved in whole epidermal. SDS-PAGE and Western blotting confirmed that the PdHsp70 protein expressed efficiently in Escherichia coli BL21. These findings have implied that PdHsp70 are implicated in different regulation of regeneration and inflammation in response to injury stimulation. During the regeneration, it is involved in the formation of wound epidermis by mediating cellular protection whereas it can modulate inflammatory by activating the innate immune response. [ABSTRACT FROM AUTHOR]

The present investigation was aimed to assess the effects of imidacloprid on the survival, genetic materials, hepatic transaminase activity and histopathology of loach ( Misgurnus anguillicaudatus ). The values of LC 50 (24, 48, 72 and 96 h) of imidacloprid were 167.7, 158.6, 147.9 and 145.8 mg/L, respectively, and the safety concentration was 42.55 mg/L. The erythrocyte micronuclei assays and the comet assay results showed that imidacloprid had genetic toxic effect on the loach erythrocytes. To assess the physiological and biochemical damage caused by imidacloprid, the activities of hepatic glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) were measured and their values declined in treatment groups. Histological examination of testis revealed that imidacloprid treatment resulted in disorganized lobules and cysts structures. In the present work, we also investigated the joint toxicity of pesticides commonly used in paddy fields (imidacloprid and lambda-cyhalothrin) on M. anguillicaudatus , and confirmed that a synergistic effect existing in the binary mixtures. The results of our study provide relevant and comparable toxicity information that are useful for safety application of pesticides. [ABSTRACT FROM AUTHOR]

Pesticides and other chemicals at environmental concentrations often have detrimental effects. Many aquatic species are particularly threatened because of their susceptibility and also because water environment are often polluted. This study preliminarily evaluated the toxicity effect of dichlorvos (DDVP) on loach (Misgurnus anguillicaudatus) using the methods of micronucleus (MN) test, hepatase activity and comet assay. The tested results showed that indeed very little DDVP had strong toxicity effect on loach and its 50% lethal concentration (LC50) at 24 h, 48 h and 96 h was 8.38 μg l−1, 7.168 μg l−1 and 6.411 μg l−1, respectively; The glutamic-pyruvic transaminase (GPT) and glutamic–oxalacetic transaminase (GOT) activity of loach liver decreased; meanwhile, the GPT and GOT activity of loach serum, the MN rate (‰) and three comet parameters of tested fish increased with the increase in the treatment concentration and treatment time of DDVP, and there was significant difference between control group and each treatment group (p < 0.05). These results suggested that DDVP residues might become toxic chemical contaminant in environment and would threaten aquatic and other organisms. [ABSTRACT FROM AUTHOR]

Abstract: Heat shock protein 60 (Hsp60) is a kind of highly conserved immunogenic molecule involved in a wide range of biochemical processes in response to external stressors. Its multifunction in regulating immune responses and modulating signal pathway interests us in investigating its role in fin regeneration that has become an excellent and interesting model for studying the molecular basis of morphogenesis. We firstly clarified basical process and crucial period of caudal fins regeneration in Paramisgurnus dabryanus by histological analysis. Then we cloned full-length cDNA of hsp60 from P. dabryanus (designated as PdHsp60) by RACE method. The cDNA contains a 124 bp 5′UTR, a 1731 bp open reading frame (ORF) encoding 576 amino acids and a 510bp 3′UTR (Accession no.: KF544774). The phylogenetic tree shows that the PdHsp60 fits within the hsp60 clade. And quantitative RT-PCR detected the PdHsp60 began to increase rapidly its expression at 1 dpa and reached its peak at 2 dpa. Next, spatial distribution analysis of PdHsp60 in fins showed that PdHsp60 located mainly in the deeper lay of regenerated epidermis when PdHsp60 expressed most. After the PdHsp60 had been cloned into the pET-32a vector, SDS-PAGE and Western blotting analysis confirmed that the PdHsp60 protein was efficiently expressed in Escherichia coli BL21. These findings have revealed that PdHsp60, a highly conserved gene related to the innate immune system and stress response during vertebrate evolution, is involved in response to wounding stimulation—in the formation of wound epidermis which occurs as the first phase of fin regeneration after fin amputation in caudal fin regeneration. [Copyright &y& Elsevier]

This study was a preliminary step to evaluate the acute toxicity of 1-methyl-3-octylimidazolium chloride ([C8mim]Cl) on loach (Misgurnus anguillicaudatus) by determining the effects on hepatic antioxidant enzyme activities and by the comet assay. The results showed that [C8mim]Cl had acute toxicity at concentrations above 20 mg L−1, inducing oxidative stress and genotoxicity on fish liver cells. In respect to enzyme activities, [C8mim]Cl induced changes in the activities of superoxide dismutase, catalase, and glutathione content the livers of fish exposed at 20–80 mg L−1. [C8mim]Cl at the same exposure level caused a remarkable increase in malondialdehyde level. The comet assay indicated that [C8mim]Cl at 20–80 mg L−1induced genotoxicity in liver cells. With increased exposure concentration and time, the two comet parameters trailing rate and tail moment were significantly increased, with significant differences (P< 0.05) observed between control group and each treatment group. The present study shows that ionic liquids can be a threat to the health of aquatic organism when accidentally released to aquatic ecosystems. [ABSTRACT FROM PUBLISHER]

Yellow River carp (Cyprinus carpio) is an economically-important freshwater fish. It is the common host of the epizootic bacterium Flavobacterium columnare, a common fish pathogen that causes columnaris disease resulting in aquacultural losses. However, information on the functions and mechanisms of the immune system of Yellow River carp infected with F. columnare is limited. Therefore, the aim of this study is to evaluate the genetic and histopathological effects of an experimentally-induced F. columnare infection in Yellow River carp. Sixty fish were divided into control (CT group) and challenged groups. The gills were collected for histological and transcriptome analysis to understand the host immune response after challenge with F. columnare. The infected fish of the IF group presented typical columnaris disease symptoms and higher mortality, as well as histological changes. However, some challenged fish showed asymptomatic infection (IC group). Additionally, there were 1776 significant differentially-expressed genes (DEGs) between the IC and CT groups, 1853 DEGs between the IF and CT groups, and 1836 DEGs between the IF and IC groups, All the DEGs were classified into three gene ontology categories, which were allocated to 158 KEGG pathways. Moreover, immune-related genes were confirmed by qRT-PCR. we quantified the level of IL-1, IL-6, TNF-α and IL-8 by ELISA. The results showed the highest expression levels of inflammatory cytokines as well as stress proteins and the adhesion molecules in the lF group, which may contribute to severe infection, and a higher case fatality rate, while the high expression of chemokines, costimulatory molecules and the up regulation of antigen presentation function could help the carp resist F. columnare infection. • The pathological changes of carp gills were caused by the infection of F. columnare. • Transcriptome can provide immune-related DEGs of carp infected with F. columnare. • Inflammatory cytokines may contribute to higher case fatality in the infected carp. [ABSTRACT FROM AUTHOR]

To investigate the function and evolutionary origin of the SOXE subgroup, we amplified the genomic DNA of Misgurnus anguillicaudatus using a pair of degenerate primers. Using RACE, we obtained two versions of Sox8 ( MaSox8a and MaSox8b) from M. anguillicaudatus. The overall sequence identity of the deduced amino acids from the two genes was 54.38%, with only one amino acid change in the high-mobility group domain. Southern blotting and evidence from the phylogenetic tree provided further proof for the existence of two Sox8 genes at the genomic level. This is the first evidence of two distinct Sox8 genes in Cypriniformes. Semi-quantitative and real-time quantitative PCR assays showed the expression trend of the genes was opposite in early embryonic development, and both were expressed ubiquitously in several adult tissues. The similar expression patterns indicated that MaSox8a and MaSox8b have possible overlapping functions. [ABSTRACT FROM AUTHOR]

Abstract: In this study, we identified a sex-specific DNA marker using randomly amplified polymorphic DNA (RAPD) fingerprinting in Yellow River carps (Cyprinus carpio from the Yellow River). Two hundred and twenty random primers were used in pooled DNA samples and individual DNA samples from male and female fish for RAPD fingerprinting. When using the primer S2107, a novel sex-specific PCR product was identified in all male individuals. DNA sequencing revealed that this 909 bp long DNA fragment has a low similarity to a repetitive sequence in zebrafish Danio rerio. In order to confirm the amplification results, two primers were designed within the male-specific sequence in order to amplify the sex-specific fragment from genomic DNA of male and female carps for sexing by PCR. The results indicated bands specific for males but not females. Respective results were obtained in dot blot and Southern blot hybridization experiments when using this sex-specific fragment as the probe. The sex-specific pattern was observed in 30 individuals from three separate common carp stocks, suggesting that the sequence is conserved in common carp species. However, no hybridization signals were found in grass carps Ctenopharyngodon idella, which are related to common carps. We submit to use this sex-specific fragment as a marker to rapidly and accurately identify the gender of Yellow River C. carpio. Furthermore, the sex-specific chromosome region may be characterized and used to study mechanisms of chromosome evolution in this fish species. [Copyright &y& Elsevier]

[Display omitted] • The toxicity effects of mixture and MCPA-Na were compared. • The herbicide mixtures caused a more serious lethal effect and teratogenicity. • The herbicide mixtures produced synergistic effect among embryos. • Explored the potential mechanism of synergistic effect caused by herbicide mixtures. Herbicides may pose considerable danger to non-target aquatic organisms and further threaten human health. The present investigation was aimed to assess the effects of 2-methyl-4-chlorophenoxy acetic acid (MCPA-Na) on Cyprinus carpio embryos. Embryos were exposed to six concentrations of MCPA-Na (0, 52, 54, 56, 58 and 60 mg/L) for 96 h. A series of symptoms were observed in developmental embryos during MCPA-Na exposure, including increased death, hatching inhibited and morphological deformities. Further, MCPA-Na exposure leading to a series of morphological changes (pericardial edema, tail deformation, and spine deformation) in embryos, which were consistent with modifications in the associated genes. In this work, we also investigated the joint toxicity of herbicides (MCPA-Na and cyhalofop-butyl) commonly used in paddy fields on carp embryos, using the 96 h-LC 50 of herbicides (59.784 mg/L MCPA-Na and 1.472 mg/L cyhalofop-butyl) and confirmed that a synergistic effect existing in the binary mixtures. [ABSTRACT FROM AUTHOR]

Dechlorane Plus (DP) is a widely used chlorinated flame retardant, which has been extensively detected in the environment. Although DP content in the surface water is low, it can pose a continuous exposure risk to aquatic organisms due to its strong bioaccumulation. Considering that the related studies on the toxicity mechanism of DP exposure are limited, the effect of DP on carp embryo development was evaluated. In the present work, carp embryos were exposed to different concentrations (0, 30, 60, and 120 μg/L) of DP at 3 h post-fertilization (hpf). The expression levels of neural and skeletal development-associated genes, such as sox2, sox19a, Mef2c and BMP4 , were detected with quantitative PCR, and the changes in different developmental toxicity endpoints were observed. Our results demonstrated that the expression levels of sox2, sox19a, Mef2c and BMP4 were significantly altered and several developmental abnormalities were found in DP-exposed carp embryos, such as DNA damage, increased mortality rate, delayed hatching time, reduced hatching rate, decreased body length, and increased morphological deformities. In addition, the activities of reactive oxygen species and malondialdehyde were remarkably higher in 60 and 120 μg/L DP exposure groups than in control group. These results suggest that DP can exhibit a unique modes of action, which lead to aberration occurrence in the early development stage of common carps, which may be related to some gene damage and oxidative stress. Besides, the parameters evaluated here can be used as tools to access the environmental risk for biota and humans exposed to DP. Image 1 • DP has embryotoxic effects on common carp. • DP could delay embryo development and even cause embryo death. • DP exposure could induce DNA damage and oxidative stress. • ROS and MDA may play an important role in DP-induced developmental deformity. [ABSTRACT FROM AUTHOR]

The occurrence and accumulation of microplastics in wildlife and humans have become a serious global scale concern over the last decade. To evaluate the potential toxic effects of PVC (polyvinyl chloride) microplastics in freshwater fish larvae, we conducted chronic 30-day and 60-day dietary exposure using Cyprinus carpio var. larvae. We exposed the larvae to four treatments with different microplastic concentrations (10%, 20%, and 30%) using food rationed diets, in conjunction with a non-plastic control. The results indicated that microplastics significantly inhibited weight gain and growth under all PVC treatments, compared to the control group. SOD (superoxide dismutase) and CAT (catalase) activities were analyzed, and an inverse relationship between them was observed. The activities of GPx (glutathione peroxidase) initially ascended and then descended with increased PVC concentrations following 30 days of exposure. A dose dependent downtrend was observed after 60 days of exposure. Malondialdehyde (MDA) levels were significantly reduced upon exposure to different concentrations of microplastics in various tissues. Altered antioxidant-related gene expression was observed in the livers of larvae exposed to the PVC microplastics. The transcription of CYP1A and GSTa initially increased, and then decreased under higher microplastics concentrations following 30 days of exposure. Furthermore, histological studies revealed cytoplasmic vacuolation in the liver under exposure to 20% and 30% microplastics. This investigation provided basic toxicological data toward elucidating and quantifying the impacts of PVC microplastics on freshwater organisms. Unlabelled Image • PVC microplastics inhibited weight gain and body length growth of carp larvae. • Antioxidant activities were altered following PVC microplastics exposure. • PVC microplastics influenced the expression levels of CYP1A , GSTpi , and GSTa. • Histological changes occurred in the livers of fish exposed to PVC microplastics. [ABSTRACT FROM AUTHOR]