Whole-Genome Identification and Characterization of the DKK Gene Family and Its Transcription Profiles: An Analysis of the Chinese Soft-Shell Turtle (Pelodiscus sinensis).

The DKK family is a canonical small family of WNT antagonists. Though recent studies have suggested that the DKK gene family may be involved in sex differentiation in Pelodiscus sinensis, there are still a lot of things about the DKK gene family that we do not know. In this study, we used bioinformatics methods to identify members of the DKK gene family in P. sinensis and analyzed their phylogeny, covariance, gene structure, structural domains, promoter conserved sites, signal peptides, gonadal transcription factors, transcriptional profiles, and tissue expression profiles. Additionally, qRT-PCR results were utilized for the validation and preliminary investigation of the function of the DKK gene family in P. sinensis. The results showed that the DKK gene family is divided into six subfamilies, distributed on six different chromosomal scaffolds containing different gene structures and conserved motifs with the same structural domains, and all of the members were secreted proteins. Our transcriptional profiling and embryonic expression analysis showed that DKKL1 and DKK4 were significantly expressed in the testes, whereas DKK1 and DKK3 were significantly upregulated in the ovaries. This suggests a potential function in sex differentiation in P. sinensis. Our results may provide a basic theoretical basis for the sex differentiation process in P. sinensis.

Identification of Sex-Specific Markers and Candidate Genes Using WGS Sequencing Reveals a ZW-Type Sex-Determination System in the Chinese Soft-Shell Turtle (Pelodiscus sinensis).

Male and female Chinese soft-shelled turtles (Pelodiscus sinensis) have sex-dimorphic growth patterns, and males have higher commercial value because of their larger size and thicker calipash. Thus, developing sex-specific markers is beneficial to studies on all-male breeding in P. sinensis. Here, we developed an accurate and efficient workflow for the screening of sex-specific sequences with ZW or XY sex determination systems. Based on this workflow, female and male P. sinensis reference genomes of 2.23 Gb and 2.26 Gb were obtained using de novo assembly. After aligning and filtering, 4.01 Mb female-specific sequences were finally identified. Subsequently, the seven developed sex-specific primer pairs were 100% accurate in preliminary, population, and embryonic validation. The presence and absence of bands for the primers of P44, P45, P66, P67, P68, and P69, as well as two and one bands for the PB1 primer, indicate that the embryos are genetically female and male, respectively. NR and functional annotations identified several sex-determining candidate genes and related pathways, including Ran, Eif4et, and Crkl genes, and the insulin signaling pathway and the cAMP signaling pathway, respectively. Collectively, our results reveal that a ZW-type sex-determination system is present in P. sinensis and provide novel insights for the screening of sex-specific markers, sex-control breeding, and the studies of the sex determination mechanism of P. sinensis.

Comparative genomic survey and functional analysis of DKKL1 during spermatogenesis in the Chinese soft-shelled turtle (Pelodiscus sinensis).

A feature of the Chinese soft-shelled turtle (Pelodiscus sinensis) is seasonal spermatogenesis; however, the underlying molecular mechanism is not well clarified. Here, we firstly cloned and characterized P. sinensis DKKL1, and then performed comparative genomic studies, expression analysis, and functional validation. P. sinensis DKKL1 had 2 putative N-glycosylation sites and 16 phosphorylation sites. DKKL1 also had classic transmembrane structures that were extracellularly localized. DKKL1's genetic distance was close to turtles, followed by amphibians and mammals, but its genetic distance was far from fishes. DKKL1 genes from different species shared distinct genomic characteristics. Meanwhile, they were also relatively conserved among themselves, at least from the perspective of classes. Notably, the transcription factors associated with spermatogenesis were also identified, containing CTCF, EWSR1, and FOXL2. DKKL1 exhibited sexually dimorphic expression only in adult gonads, which was significantly higher than that in other somatic tissues (P < 0.001), and was barely expressed in embryonic gonads. DKKL1 transcripts showed a strong signal in sperm, while faint signals were detected in other male germ cells. DKKL1 in adult testes progressively increased per month (P < 0.05), displaying a seasonal expression trait. DKKL1 was significantly downregulated in testes cells after the sex hormones (17ß-estradiol and 17α-methyltestosterone) and Wnt/ß-catenin inhibitor treatment (P < 0.05). Likewise, the Wnt/ß-catenin inhibitor treatment dramatically repressed CTCF, EWSR1, and FOXL2 expression. Conversely, they were markedly upregulated after the 17ß-estradiol and 17α-methyltestosterone treatment, suggesting that the three transcription factors might bind to different promoter regions, thereby negatively regulating DKKL1 transcription in response to the changes in the estrogen and androgen pathways, and positively controlling DKKL1 transcription in answer to the alterations in the Wnt/ß-catenin pathway. Knockdown of DKKL1 significantly reduced the relative expression of HMGB2 and SPATS1 (P < 0.01), suggesting that it may be involved in seasonal spermatogenesis of P. sinensis through a positive regulatory interaction with these two genes. Overall, our findings provide novel insights into the genome evolution and potential functions of seasonal spermatogenesis of P. sinensis DKKL1.

Sex-specific metabolic dysregulation in digestive glands of green mussels following exposure to triazophos.

As a ubiquitous environmental pollutant in China, triazophos (TP) is known to have neurotoxicity, oxidative stress, and reproductive toxicity to mussels. To investigate the molecular mechanisms of TP toxicity, metabolic changes in the digestive glands of Perna viridis in different sexes were examined after treated with 35 µg/L TP. Notably, 158 significant different metabolites (SDMs) were detected in TP-treated mussels and more than half of the SDMs were lipids and lipid-like molecules, which suggested that TP disturbed the lipid metabolism of P. viridis. In addition, metabolites associated with neurotoxicity and reproductive disturbance were also detected in female and male mussels. Moreover, a larger number of SDMs were found in male mussels (120 SDMs) than females (99 SDMs), and 60 common metabolites exhibited consistent variation tendency and similar magnitude in both sexes. The metabolic alternations in female and male mussels displayed similar protective mechanisms and also sex-specific responses, male mussels were more sensitive to TP exposure. This research provided new data about the molecular mechanisms of TP toxicity and the gender specific changes in mussels after treated by chemicals.

Source apportionment of nitrate in the Pearl River Estuary using δ15N and δ18O values and isotope mixing model.

The mitigation of eutrophication in the Pearl River Estuary (PRE) has encountered numerous challenges in regards to source control. Herein, the isotope mixing model (SIAR) was used to quantify the primary nitrate sources in the PRE. The results showed that the nitrate levels were significantly higher in the high-flow season than in the low-flow season. Meanwhile, we found the most important nitrate sources were manure and sewage during the high-flow season, with a contribution ratio of 47 % in the low salt area (LSA) and 29 % in the high salt area (HSA). During the low-flow season, the primary nitrate sources were identified as reduced nitrogen fertilizer in the LSA and manure and sewage in the HSA, which accounted for 52 % and 44 %, respectively. Furthermore, we also suggest that a feasible measure might be to control the pollution caused in the PRE by manure and sewage as well as reduced nitrogen fertilizer.

Genome-wide identification, evolution and expression analysis of bone morphogenetic protein (BMP) gene family in chinese soft-shell turtle (Pelodiscus sinensis).

Introduction: Bone morphogenetic proteins (BMPs) play a crucial role in bone formation and differentiation. Recent RNA-Seq results suggest that BMPs may be involved in the sex differentiation of P. sinensis, yet more relevant studies about BMPs in P. sinensis are lacking. Methods: Herein, we identified BMP gene family members, analyzed the phylogeny, collinear relationship, scaffold localization, gene structures, protein structures, transcription factors and dimorphic expression by using bioinformatic methods based on genomic and transcriptomic data of P. sinensis. Meanwhile, qRT-PCR was used to verify the RNA-Seq results and initially explore the function of the BMPs in the sex differentiation of P. sinensis. Results: A total of 11 BMP genes were identified, 10 of which were localized to their respective genomic scaffolds. Phylogenetic analysis revealed that BMP genes were divided into eight subfamilies and shared similar motifs ("WII", "FPL", "TNHA", "CCVP", and "CGC") and domain (TGF-ß superfamily). The results of the sexually dimorphic expression profile and qRT-PCR showed that Bmp2, Bmp3, Bmp15l, Bmp5, Bmp6 and Bmp8a were significantly upregulated in ovaries, while Bmp2lb, Bmp7, Bmp2bl and Bmp10 were remarkable upregulated in testes, suggesting that these genes may play a role in sex differentiation of P. sinensis. Discussion: Collectively, our comprehensive results enrich the basic date for studying the evolution and functions of BMP genes in P. sinensis.

Expression and Characterization of the Spats1 Gene and Its Response to E2/MT Treatment in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis).

Spats1 (spermatogenesis-associated, serinerich 1) has been characterized as a male-biased gene which acts an important role in the germ cell differentiation of mammals. Nevertheless, the function of Spats1 in the Chinese soft-shelled turtle (P. sinensis) has not yet been reported. To initially explore the expression of Spats1 in P. sinensis and its response to sex steroid treatment, we cloned the CDS of Spats1 for the first time and analyzed its expression profile in different tissues, including the testes in different seasons. The Spats1 cDNA fragment is 1201 base pairs (bp) in length and contains an open reading frame (ORF) of 849 bp, which codes for 283 amino acids. Spats1 mRNA was highly expressed in the testes (p < 0.01) and barely detectable in other tissues. In P. sinensis, the relative expression of Spats1 also responsive to seasonal changes in testis development. In summer (July) and autumn (October), Spats1 gene expression was significantly higher in the testes than in other seasons (p < 0.05). Spats1 mRNA was found to be specifically expressed in germ cells by chemical in situ hybridization (CISH), and it was mainly located in primary spermatocytes (Sc1), secondary spermatocytes (Sc2) and spermatozoa (St). Spats1 expression in embryos was not significantly changed after 17α-methyltestosterone (MT)and 17ß-estradiol (E2) treatment. In adults, MT significantly induced Spats1 expression in male P. sinensis. However, the expression of Spats1 in testes was not responsive to E2 treatment. In addition, the expression of Spats1 in females was not affected by either MT or E2 treatment. These results imply that Spats1 is a male-specific expressed gene that is mainly regulated by MT and is closely linked to spermatogenesis and release in P. sinensis.

Whole-Transcriptome Analysis Identifies Gender Dimorphic Expressions of Mrnas and Non-Coding Rnas in Chinese Soft-Shell Turtle (Pelodiscus sinensis).

In aquaculture, the Chinese soft-shelled turtle (Pelodiscus sinensis) is an economically important species with remarkable gender dimorphism in its growth patterns. However, the underlying molecular mechanisms of this phenomenon have not been elucidated well. Here, we conducted a whole-transcriptome analysis of the female and male gonads of P. sinensis. Overall, 7833 DE mRNAs, 619 DE lncRNAs, 231 DE circRNAs, and 520 DE miRNAs were identified. Some "star genes" associated with sex differentiation containing dmrt1, sox9, and foxl2 were identified. Additionally, some potential genes linked to sex differentiation, such as bmp2, ran, and sox3, were also isolated in P. sinensis. Functional analysis showed that the DE miRNAs and DE ncRNAs were enriched in the pathways related to sex differentiation, including ovarian steroidogenesis, the hippo signaling pathway, and the calcium signaling pathway. Remarkably, a lncRNA/circRNA-miRNA-mRNA interaction network was constructed, containing the key genes associated with sex differentiation, including fgf9, foxl3, and dmrta2. Collectively, we constructed a gender dimorphism profile of the female and male gonads of P. sinensis, profoundly contributing to the exploration of the major genes and potential ncRNAs involved in the sex differentiation of P. sinensis. More importantly, we highlighted the potential functions of ncRNAs for gene regulation during sex differentiation in P. sinensis as well as in other turtles.

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To understand the population characteristics of Collichthys lucidus, an important economic fish in the Pearl River Estuary, the biological characteristics and resource density distribution characteristics of C. lucidus were preliminarily analyzed using bottom trawling by cruises conducted in each spring and autumn during 2017 and 2020. The results showed that the body length and weight of C. lucidus ranged between 22-168 mm and 0.23-103.11 g, respectively. Female individuals were larger than the male ones. The length of sexually mature individuals intensively ranged between 90 mm and 140 mm. Neither of them evidenced the earlier of sexually maturity nor the minimizer of dominant group. The population of C. lucidus in Pearl River estuary still developed in safe status in all, but its habitat downgraded than in 1988, as indicated by the fact that the allometric growth factor (b=2.9057) of the body length to body weight had no significant annual variations, but the conditional factor (a=3.029×10-5) was drama-tically decreased than in 1988. The population was at a state of overexploitation due to the estimated exploitation rate of 0.67. The resource density averaged 77.73 kg·km-2, showing a pattern of higher in the middle and west than in the east and relatively uniform of latitudinal distribution. The four high densities of sampling zones suggested that the zone around Nansha Port was probably the core of spawning ground of C. lucidus. Considering the annual average resource density in 2017-2020 sharply decreased by 93.5% than in 1980 to 1982, it was pressing to establish the protection zone in spawning ground in spring to protect the recruiting and spawning stocks of C. lucidus population.

Tetrabromobisphenol A and hexabromocyclododecanes in sediments from fishing ports along the coast of South China: Occurrence, distribution and ecological risk.

Tetrabromobisphenol A (TBBPA) and hexabromocyclododecanes (HBCDDs) have attracted extensive attention due to their strong persistence and toxicity. However, little has been known about their pollution status in fishing ports, which are typical sinks of land-sourced pollutants. In this study, we investigated the occurrence, distribution and ecological risk of TBBPA and HBCDDs in sediments from fishing ports along the coast of South China. The concentrations of TBBPA and ΣHBCDD (sum of α-, ß-, and γ-HBCDD) in the fishing-port sediments were in the ranges of 0.02-21.5 ng/g dw and 1.06-14.1 ng/g dw, respectively. γ-HBCDD was the predominant diastereoisomer in most fishing-port sediments. The enantiomeric analysis indicated a preferential enrichment of (-)-enantiomers for α-, ß-, and γ-HBCDD. The geographical location of fishing ports is a significant determinant of distribution for TBBPA and HBBCDs. The concentrations of TBBPA and HBCDDs in fishing-port sediments were strongly associated with local population density, but weakly correlated with total organic carbon content of the sediment. The mass inventories of TBBPA and ΣHBCDD were estimated to be 77.0 ng/cm2 and 141 ng/cm2, respectively. The ecological risk assessment demonstrated that TBBPA and HBCDDs in fishing-port sediments exhibited low risks to marine organisms. This study contributes to the understanding pollution situation of fishing ports, and provides a reference for environmental safety assessment and environmental pollution control.

Chromosome-level genome assembly of Asian yellow pond turtle (Mauremys mutica) with temperature-dependent sex determination system.

Knowledge of sex determination has important implications in physiology, ecology and genetics, but the evolutionary mechanisms of sex determination systems in turtles have not been fully elucidated, due to a lack of reference genomes. Here, we generate a high-quality genome assembly of Asian yellow pond turtle (Mauremys mutica) using continuous long-read (PacBio platform), Illumina, and high-throughput chromatin conformation capture (Hi-C) technologies. The M. mutica haplotype has a genome size of 2.23 Gb with a contig N50 of 8.53 Mb and scaffold N50 of 141.98 Mb. 99.98% sequences of the total assembly are anchored to 26 pseudochromosomes. Comparative genomics analysis indicated that the lizard-snake-tuatara clade diverged from the bird-crocodilian-turtle clade at approximately 267.0-312.3 Mya. Intriguingly, positive selected genes are mostly enriched in the calcium signaling pathway and neuroactive ligand-receptor interaction, which are involved in the process of temperature-dependent sex determination. These findings provide important evolutionary insights into temperature-dependent sex determination system.

Identification and functional analysis of miRNAs in skeletal muscle of juvenile and adult largemouth bass, Micropterus salmoides.

MicroRNAs (miRNAs) are considered key regulators to post-transcriptionally regulate gene expression affecting multiple biological activities. However, the developmental process of fish skeletal muscles is regulated by complicated molecular mechanism that has not been completely well-described. In this study, two small RNAs libraries from skeletal muscle of juvenile as well as adult largemouth bass (LMB) were obtained and sequenced using deep sequencing to investigate the development-related miRNAs. We identified an overall number of 486 already recognized miRNAs in addition to 43 novel miRNAs. Comparison of two different skeletal muscle development stages led to the identification of 220 differently expressed miRNAs between juvenile and adult LMB containing 116 up-regulated as well as 104 down-regulated miRNAs. Of them, confirmation of some differently expressed miRNAs was performed via a stem-loop qRT-PCR, which exhibited differently expressed level in juvenile and adult LMB. Furthermore, GO and KEGG enrichment analyses of targets of differently-expressed miRNAs were carried out. Additionally, the analysis of miRNAs-targets interaction network showed that miR-181b-5p_R-1, miR-725 and miR-103 as the nodal miRNAs has over 20 target genes. Moreover, miR-103 could bind the 3'-UTR of actr8, which was validated via dual-luciferase reporter assay. It has been reasonably hypothesized that miR-103 may play a crucial role, which regulate skeletal muscle development of LMB. The present study provides the first identification of miRNA expression profiles at two different skeletal muscle development stages in LMB. Results may be valuable in interpreting the regulatory role miRNAs plays in the growth and developmental process of skeletal muscle and its possible use in LMB breeding.

Transcriptome analysis reveals sex-specific alterations in gonads of green mussel exposed to organophosphorus insecticide triazophos.

Triazophos (TP) is a widespread pollutant in aquatic environments. A sex-specific metabolic response in green-lipped mussel Perna viridis to TP exposure was observed in our previous study, and this led us to investigate the mechanisms associated with its toxicity. P. viridis were subjected to chronic exposure (15 days) to TP at 35 µg/L to compare the sex-biased transcriptomic profiles in the gonads of male and female mussels. We identified 632 differentially expressed genes (DEGs) (348 up-regulated and 284 down-regulated) in TP-exposed males, and only 61 DEGs (9 up-regulated and 52 down-regulated) in TP-exposed females. Many DEGs were found to be involved in the nervous, reproductive endocrine, oxidative stress, and immune systems of P. viridis. Additionally, enzymatic activity analysis indicated TP induced neurotoxic effects and oxidative damage to the mussels. Our results demonstrate that the stress response and molecular mechanisms of TP toxicology are different between female and male mussels.

Oyster arsenic, cadmium, copper, mercury, lead and zinc levels in the northern South China Sea: long-term spatiotemporal distributions, combined effects, and risk assessment to human health.

Estuarine and coastal ecosystems are often considered vulnerable due to the complex biogeochemical processes and the human disturbances through a variety of pollution. Among environmental contaminants, heavy metals in estuarine and coastal ecosystems have been of increasing concern in environmental conservation. Long-term exposure to heavy metal contamination, mainly through food and water, could be harmful to human health. It is therefore critical to understand the quantitative comparisons and combined effects of different heavy metals in common seafood species, such as oysters. This work studied the long-term spatiotemporal trends and health risk assessment of oyster arsenic (As), cadmium (Cd), copper (Cu), mercury (Hg), lead (Pb), and zinc (Zn) levels in the coastal waters of northern South China Sea. Cultured oysters (Crassostrea rivularis) from 23 estuaries and harbors in the coastal areas of northern South China Sea in 1989-2015 were analyzed for the spatiotemporal trends of the six heavy metal levels. Metal pollution index (MPI), target hazard quotient (THQ), and hazard index (HI) were used for quantifying the exposure of the six heavy metals to human health through oyster consumption. Principal component analysis (PCA) was used for assessing the relative importance of the six metals in oyster heavy metal distribution patterns in the northern South China Sea. Overall, the As, Cd, Cu, Hg, Pb, and Zn levels in oysters from the northern South China Sea generally declined from 1989 to 2015, stayed relatively high (MPI = 2.42-3.68) during 1989-2000, gradually decreased since 2000, and slightly increased after 2010. Oyster heavy metal levels were highest in the Pearl River Estuary (MPI = 1.20-5.52), followed by west Guangdong and east Guangdong, Guangxi, and Hainan coastal waters. This pattern is probably because economics and industry around the Pearl River Estuary have been growing faster than the other areas of this work in the recent two decades, and it should be taken as a hotspot for the monitoring of seafood safety in southern China. Principal component analysis indicated that Cu, Zn, and Cd were the most important metals in the long-term distributions of oyster heavy metal levels in the northern South China Sea. Health risk assessment suggested that the risk of the six heavy metals exposure through oyster consumption were relatively high during 1989-2005 (THQ = 1.01-5.82), significantly decreased since 2005 (THQ < 1), and slightly increased after 2010.

Contamination and ecological risks of steroid metabolites require more attention in the environment: Evidence from the fishing ports.

Due to strong endocrine disrupting effects, steroids in the environment have attracted substantial attention, with studies mostly focusing on the parent steroids. Here, we conducted the first investigation on the contamination profiles, possible sources, mass inventories, and ecological risks of 27 steroids and their metabolites in 15 typical fishing ports in Southeast China. Twelve steroids were detectable in the sediment samples with the total mean concentrations of 4.6-35 ng/g. High proportions of steroid metabolites were measured in the sediments and five metabolites were newly observed. Untreated municipal sewage and aquaculture wastes constitute the possible steroid sources in the studied fishing ports. The total inventories of steroids in fishing ports ranged from 2.1-16 mg/m2, with their metabolites being important contributors. The ecological risk analysis indicated high risks across all sampling sites mainly due to the contributions of parent steroids. Furthermore, our results found that progesterone is an acceptable chemical indicator for various steroids in sediments. This study provides the first evidence of steroid metabolites in the marine environment, calling for more studies in environmental behavior and ecotoxicology of steroid metabolites.

The complete mitochondrial genome of Pangasianodon hypophthalmus (Sauvage 1878) (Siluriformes, Pangasiidae).

The striped catfish, Pangasianodon hypophthalmus (Sauvage 1878), belongs to the family Pangasiidae, is an important economic freshwater species. We determined the complete mitogenome of the Pangasianodon hypophthalmus through sanger method. The complete mitochondrial genome of Pangasianodon hypophthalmus was a circular molecule of 16,469 bp with a total GC content of 44% which contains 37 genes, including 13 protein-coding genes, 2 rRNA genes, and 22 tRNAs genes. Phylogenetic analysis showed that Pangasianodon hypophthalmus clustered together with other Pangasianodon species and was closely related to Pangasius larnaudii, both species belonged to Pangasianodon genus.

Sex-specific effects of triphenyltin chloride (TPT) on thyroid disruption and metabolizing enzymes in adult zebrafish (Danio rerio).

Although organotin compounds are known to disturb thyroid signaling and antioxidant defense system, the sex-differences underlying these effects of triphenyltin chloride (TPT) in fish remain unclear. To understand these differences, adult zebrafish (Danio rerio) were exposed to different concentrations of TPT (0, 10, 100, or 1000 ng/L) for 28 days. Female zebrafish exposed to TPT showed significantly increased thyroxine (T4) content and decrease triiodothyronine (T3) content, possibly due to downregulation of deiodinase (dio2) and uridine diphosphate glucuronosyl transferase (ugt1ab). However, decreased T4 and T3 contents in male zebrafish accompanied with upregulation of dio1, dio2 and ugt1ab. TPT exposure can lead to sex-specific thyroid disruption in adult zebrafish via alterations the Hypothalamus-pituitary-thyroid-liver axis. In addition, the gene expression levels of metabolizing enzymes, such as cyp1b, cyp1c, gpx1a, or sult1st1 were also to vary in a sex-dependent manner in adult zebrafish liver. Downregulation of cyp19a and cyp19b and decreased 17ß-estradiol (E2) contents were detected in both female and male zebrafish. Therefore, a sex-specific of thyroid disruption response after TPT exposure was observed in adult zebrafish, possibly due to inherent in female or males detoxifying enzyme capacities.

Oyster copper levels in the northern South China Sea from 1989 to 2015: spatiotemporal trend detection and human health implications.

Coastal heavy metal pollution has become an important topic for seafood safety and marine environmental protection. Unlike toxic heavy metals such as cadmium or chromium, copper is essential for oysters' growth but can inhibit their immune response to exotic stress when going above normal levels. Oysters with high copper levels can easily accumulate and transfer abnormal amounts of copper to upper trophic levels, and generate health risks for humans. This study investigated the spatiotemporal variability and health risk of copper levels in cultured oysters (Crassostrea rivularis) sampled from 23 harbors, bays, or estuaries along the northern South China Sea during 1989-2015. Overall, oyster copper concentrations in the study area ranged from 0.9 to 1897.0 µg/g wet weight with a mean of 210.0 (± 143.6) µg/g and a median of 89.3 µg/g. Although oyster copper levels in the southern China provinces of Guangdong, Guangxi, and Hainan showed an overall decrease during 1989-2015, they stayed relatively low since 1996 and increased slightly after 2010. Oyster copper levels in Guangdong were significantly higher than in Hainan and Guangxi. In Guangdong, oyster copper levels were highest in the Pearl River Estuary, followed by west Guangdong and east Guangdong. The health risk of copper exposure through oyster consumption increased in 2011-2015 compared with in 2006-2010. It is recommended that the human daily intake of cultured oysters in the study area should be reduced by half to minimize copper exposure. This study suggested that copper is one of the most important heavy metal contaminants in coastal and estuarine ecosystems of the northern South China Sea.

Effects of di-n-butyl phthalate on gills- and liver-specific EROD activities and CYP1A levels in juvenile red snapper (Lutjanus argentimaculatus).

Di-n-butyl phthalate, one of the most easily detected pollutants of phthalate esters in the environment, has been added to the priority list of hazardous substances by many countries. As one of low molecular weight phthalates, Di-n-butyl phthalate may have a great adverse potency on various aquatic organisms. In this study, the juvenile red snapper, Lutjanus argentimaculatus, was exposed to the concentrations of Di-n-butyl phthalate (20 µg L-1, 100 µg L-1 and 500 µg L-1) for 15 days. EROD activities and CYP1A levels were measured in liver and gill tissues. In gills, the similar effect has been found to inhibit or induce EROD activities and CYP1A levels, and there existed a good correlation between them. Whereas in the case of the liver, a moderate correlation was observed between EROD activities and CYP1A levels, which was mainly due to the inhibited EROD activities and the CYP1A levels with no significant difference by day 15. In conclusion, this study revealed the similar and different effects of cytochrome P450 enzymes on fish in the time-, concentration-, and tissue-dependent Di-n-butyl phthalate exposure. Furthermore, as the adverse effects indicated between CYP1A levels and EROD activities, metabolic mechanisms of phthalates in different tissues should be highly emphasized in future studies.

Transcriptome analysis of acute exposure of the Manila clam, Ruditapes philippinarum to perfluorooctane sulfonate (PFOS).

Perfluorooctane sulfonate (PFOS) is an increasingly important environmental pollutant, which has been detected almost everywhere in the environment. Despite the widespread presence of PFOS, much less notice is taken of its toxicology effects on marine bivalves. Thus, the transcriptome response to PFOS treatment (nominal concentration of 20 mg/L) in hepatopancreas of a sentinel organism, Ruditapes philippinarum was examined. Compared with the control group, 32,149 unigenes were up-regulated and 26,958 unigenes down-regulated. Notably, significant gene expression changes were found in carbohydrate metabolism, energy metabolism, amino acid metabolism, lipid metabolism and protein biosynthesis, indicating the metabolic disruptions caused by PFOS in R. philippinarum. Additionally, numerous other differentially expressed genes were involved in immune system, antioxidant defense system and detoxification metabolism. In summary, transcriptome profiling of R. philippinarum after exposure to PFOS provided molecular support for our current understanding of the detrimental toxicity of PFOS on marine bivalves.