Comparison of three palm tree peroxidases expressed by Escherichia coli: Uniqueness of African oil palm peroxidase.

Palm tree peroxidase has greater catalytic activity, stability and broad application prospects in comparison with horseradish peroxidase. However, slow growth, ecological destruction and high costs prohibit isolation of native peroxidases directly from palm trees. Bioreactor production of palm tree peroxidases would therefore be preferred to overcome such production limitations. Comparison of different recombinant glycan-free palm tree peroxidases would allow understanding the criticality of total glycans to the functions and characteristics. In the present study, African oil palm tree peroxidase expressed by Escherichia coli showed similar stability and 30-100-fold greater activity than that of recombinant royal palm tree peroxidases, but both of their comprehensive indexes were superior to the commercial, native horseradish peroxidase. Recombinant Chamaerops excelsa peroxidase showed no activity possibly due to incorrect protein folding. The results confirmed that recombinant expression by E. coli is potentially an effective means to obtain a mass of palm peroxidases with high activity and stability.

Comparative evaluation of five protocols for protein extraction from stony corals (Scleractinia) for proteomics.

Corals especially the reef-building species are very important to marine ecosystems. Proteomics has been used for researches on coral diseases, bleaching and responses to the environment change. A robust and versatile protein extraction protocol is required for coral proteomics. However, a comparative evaluation of different protein extraction protocols is still not available for proteomic analysis of stony corals. In the present study, five protocols were compared for protein extraction from stony corals. The five protocols were TRIzol, phenol-based extraction (PBE), trichloroacetic acid (TCA)-acetone, glass bead-assisted extraction (GBAE) and a commercially available kit. PBE, TRIzol and the commercial kit were more robust for extracting proteins from stony corals. The protein extraction efficiency and repeatability, two dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) were employed to evaluate the protocols. The results indicated that PBE protocol had the better protein extraction efficiency than the others. Protein extraction coverage varied among the procedures. Each protocol favored for certain proteins. Therefore, it is very important for coral proteomic analysis to select a suitable protein protocol upon the experimental design. In general, PBE protocol can be the first choice for extracting proteins from stony corals.

An integrated metabolomic and proteomic study of toxic effects of Benzo[a]pyrene on gills of the pearl oyster Pinctada martensii.

Benzo[a]pyrene (BaP) is one of the most important polycyclic aromatic hydrocarbons (PAHs), which are widely present in the marine environment. Because of its teratogenic, mutagenic, and carcinogenic effects on various organisms, the toxicity of BaP is of great concern. In this study, we focused on the toxic effects of BaP (1 µg/L and 10 µg/L) on gills of the pearl oyster Pinctada martensii using combined metabolomic and proteomic approaches. At the metabolome level, the high concentration of BaP mainly caused abnormal energy metabolism, osmotic regulation and immune response marked by significantly altered metabolites in gills. At the proteome level, both concentrations of BaP mainly induced signal transduction, transcription regulation, cell growth, stress response, and energy metabolism. Overall, the research demonstrated that the combination of proteomic and metabolomic approaches could provide a significant way to elucidate toxic effects of BaP on P. martensii.

Occurrence and distribution of Polycyclic aromatic hydrocarbons (PAHs) in seawater, sediments and corals from Hainan Island, China.

The levels of 16 US EPA priority polycyclic aromatic hydrocarbons (PAHs) were investigated in corals, ambient seawater and sediments of Hainan Island, China, using gas chromatography - mass spectrometry (GC-MS). The total PAHs (∑PAHs) concentrations ranged from 273.79 to 407.82ng/L in seawater. Besides, the concentrations of ∑PAHs in corals 333.88-727.03ng/g dw) were markedly (P < 0.05) higher than ambient sediments 67.29-196.99ng/g dw), demonstrating the bioaccumulation ability of PAHs by corals. The highest concentration of ∑PAHs was detected at site S2 in Pavona decussate, which also bore the highest ∑PAHs levels in both seawater and sediments. The massive corals were more enriched with PAHs than the branching corals. Although 2 and 3-ring PAHs were predominant and accounted for 69.27-80.46% of the ∑PAHs in corals and ambient environment, the levels of high molecular weight (HMW) PAHs (4-6 ring) in corals also demonstrated their potential dangers for corals and organisms around coral reefs. Biota-sediment accumulation factor (BSAF) refers to an index of the pollutant absorbed by aquatic organisms from the surrounding sediments. The poor correlation between log BSAF and log Kow (hydrophobicity) indicated that PAHs in corals maybe not bioaccumulate from the ambient sediments but through pathways like absorbing from seawater, symbiosis, and feeding. Based on our data, long-term ecological monitoring in typical coral reef ecosystems combined with ecotoxicological tests of PAHs on corals is necessary to determine the impacts of PAHs on coral reefs.

Effects of pyrene exposure on immune response and oxidative stress in the pearl oyster, Pinctada martensii.

Pyrene is a polycyclic aromatic hydrocarbon (PAH) commonly observed in aquatic ecosystems, which originates primarily from the incomplete combustion of fossil fuels and the use of petroleum compounds. Pyrene can cause the immune disturbance and oxidative stress, result in immunotoxicity, DNA damage, reduce reproduction significantly, and induce behavioral changes. Marine bivalves are commonly used as bioindicators for marine pollution, and hemolymph is a metabolite transfer medium for PAH pollutant. However, the vital immune indicator responses of pearl oyster Pinctada martensii hemolymph exposed to pyrene is still unclear. Thus, the immunotoxic responses of pyrene on the hemolymph of the Pinctada martensii were investigated in this study. After exposure to pyrene for 7 days, the total number of hemocytes (THC), cell membrane stability (CMS), phagocytic activity (PA) and total glutathione (GSHT) all decreased significantly. Pyrene also caused a significant increase in lipid peroxidation (LPO). Median effective concentrations (EC50) of pyrene on THC (4.5 µg L-1) and LPO (5.2 µg L-1) were lower than those for CMS (13.8 µg L-1), PA (12.1 µg L-1) and GSHT (7.2 µg L-1), which indicates that THC and LPO were more sensitive. Additionally, a clear dose-effect relationship indicated that pyrene stimulated a marked immune response, as well as oxidative stress in P. martensii, which demonstrates the subtle effects of pyrene exposure on marine invertebrates and the potential associated risk.

Comparative Studies on the Toxicokinetics of Benzo[a]pyrene in Pinctada martensii and Perna viridis.

Research on the kinetics of Benzo[a]pyrene (B[a]P) bioaccumulation in the clam Pinctada martensii and mussel Perna viridis showed that the initial rate of uptake was directly related to the PAH concentrations in the ambient environment. The uptake and depuration rate constants were different at the four B[a]P exposure levels, which indicated that the toxicokinetic rate constants mainly depended on the exposure levels of pollutants to the environment. In addition, the uptake rate constants of B[a]P were higher than the depuration rate constants in the entire experiment. The comparison demonstrated that mussels release B[a]P more rapidly than clams. The bioconcentration factors (BCFs) of B[a]P varied from 3335 to 12892 in the clam and 2373-6235 in the mussel. These findings on the bioaccumulation kinetics for petroleum hydrocarbons, in association with the critical body residue, will be valuable when choosing sensitive organisms to assess the potential ecotoxicological risk to the marine environment.

Gene cloning and expression analysis of AhR and CYP4 from Pinctada martensii after exposed to pyrene.

Pyrene, a typical polycyclic aromatic hydrocarbon, is a common pollutant in the marine environment. Polycyclic aromatic hydrocarbons initiate cellular detoxification in an exposed organism via the activation of the aryl hydrocarbon receptor (AhR). Subsequent metabolism of these xenobiotics is mainly by the cytochrome P450 enzymes of the phase I detoxification system. Full-length complementary DNA sequences from the pearl oyster Pinctada martensii (pm) encoding AhR and cytochrome P4 were cloned. The P. martensii AhR complementary DNA sequence constitutes an open reading frame that encodes for 848 amino acids. Sequence analysis indicated PmAhR showed high similarity with its homologues of other bivalve species. The cytochrome P(CYP)4 complementary DNA sequence of P. martensii constitutes an open reading frame that encodes for 489 amino acids. Quantitative real-time analysis detected both PmAhR and PmCYP4 messenger RNA expressions in the mantle, gill, hepatapancreas and adductor muscle of P. martensii exposed to pyrene. The highest transcript-band intensities of PmAhR and PmCYP4 were observed in the gill. Temporal expression of PmAhR and PmCYP4 messenger RNAs induction was observed in gills and increased between 3 and 5 days post exposure; then returned to control level. These results suggest that messenger RNAs of PmAhR and PmCYP4 in pearl oysters might be useful parameters for monitoring marine environment pyrene pollution.

Bioaccumulation and Elimination of the Herbicide Clomazone in the Earthworms Eisenia fetida.

Acute toxicity, bioaccumulation, and elimination of herbicide clomazone in the earthworm Eisenia fetida were investigated in the different exposure systems. The LC50 values of clomazone on earthworms were 5.6 µg cm(-2) in the contact filter paper test (48 h), 174.9 mg kg(-1) (7 days) and 123.4 mg kg(-1) (14 days) in artificial soil test, respectively. Clomazone could rapidly bioaccumulate in earthworms and reached the highest concentration after 3 days exposure, with the maximum concentrations of 9.0, 35.3 and 142.3 mg kg(-1) at 10.0, 40.0 and 160.0 mg kg(-1) of clomazone, respectively. Clomazone uptake showed a good correlation with exposure concentration. After the 14th day, clomazone declined to minimum value. About 74%-80% of accumulated clomazone was eliminated within 1 day after exposed to clomazone-free soil. However, a trace amount of clomazone persisted for a relatively long time in earthworms.

Chronic exposure to low concentrations of microplastics causing gut tissue damage but non-significant changes in the microbiota of marine medaka larvae (Oryzias melastigma).

Microplastics (MPs) have become a popular research topic due to their potential ramifications on aquatic organisms. To evaluate the ecotoxicological impacts of chronic exposure to different microplastics on marine medaka larvae, we exposed medaka larvae to 200 µg/L of polyethylene (PE-200) and polylactic acid (PLA-200) microplastics for 60 days, respectively. The results indicated that both exposures had no significant effect on fish length/weight and did not result in fish mortality. Notably, the structure of intestinal microbiota was not disrupted either. However, microscopy observations of intestinal tissue suggested that exposure to MPs resulted in inflammation of the intestinal tract of fish and significant atrophy and shedding of small intestinal villus. Linear discriminant analysis Effect Size (LEfSe) showed that intestinal enrichment of Streptomyces occurred in marine medaka larvae in both MPs treatments, while the PE-200 treatment exhibited a significant enrichment. In addition, the PICRUSt2 prediction indicated significant upregulation of the Novobiocin biosynthesis function in gut microbiota in the PE-200 treatment. Overall, multi-level assessment is necessary to determine the risk of exposure of aquatic organisms to MPs.

Distribution characteristics of antibiotic resistance genes and microbial diversity in the inshore aquaculture area of Wenchang, Hainan, China.

The rapid development of marine aquaculture has led to the increased use and release of antibiotics into the marine environment, consequently contributing to the emergence of antibiotic resistance. Information on antibiotic resistance in nearshore marine aquaculture areas remains limited, and research on the microbial composition and potential hosts of antibiotic resistance genes (ARGs) in marine aquaculture areas is scarce. This study used SmartChip real-time fluorescent quantitative PCR and qPCR to quantitatively analyze 44 ARGs and 10 mobile genetic elements (MGEs) genes in 12 sampling points in the nearshore aquaculture area of Wenchang. High-throughput sequencing of 16S rRNA was used to study microbial diversity in the study area, to clarify the correlation between ARGs, MGEs, and microbial diversity, and to determine the possible sources and potential hosts of ARGs. The results showed that a total of 37 ARGs and 8 MGEs were detected in the study area. The detection rate of 9 ARGs (aac(6')-Ib(aka aacA4)-02, catA1, cmlA, cfr, sul1, sul2, sulA/folP-01, tetC, tetX) was 100 %. The absolute abundance of ARGs in the 12 sampling points ranged from 2.75 × 107 to 3.79 × 1010 copies·L-1, and the absolute abundance of MGEs was 1.30 × 105 to 2.54 × 107 copies·L-1, which was relatively high compared to other research areas. ARGs and MGEs were significantly correlated, indicating that MGEs play an important role as a mediator in the spread of ARGs. At the phylum level, Proteobacteria and Cyanobacteria were the dominant bacteria in the study area, with HIMB11 and unidentifiedChloroplast being the dominant levels, respectively. Network analysis of ARGs and microorganisms (genus level) revealed that Cognatishimia, Thalassobius, Aestuariicoccus, Thalassotalea, and Vibrio were significantly correlated with multiple ARGs and were the main potential hosts of ARGs in the nearshore waters of Wenchang.

Distribution, sources, ecological and human health risks of organic ultraviolet filters in coastal waters and beach deposits in Hainan, China.

Organic ultraviolet filters (OUVFs) are extensively incorporated into both cosmetic items and industrial products and have been commonly found in water ecosystems. This study aims to examine the environmental levels, sources, ecological and human health risks of 14 commonly used OUVFs both in coastal water and beach deposit samples collected from the nearshore regions of Hainan Island and the South China Sea. This is first study highlighting the contamination of OUVFs in Hainan Island and utilizing economic and tourism data to confirm the potential source of OUVF pollution in costal aquatic and coastal ecosystem. Along the coastal tourist regions of Hainan Island, the median concentrations in coastal waters and beach deposits of these OUVFs fall within the range from 1.2 to 53.2 ng/L and 0.2-17.0 ng/g dw, respectively. In coastal water and beach deposit, the concentration of BP-3 was the highest, with median concentrations of 53.2 ng/L and 17.0 ng/g dw, respectively. Regarding human health risks, the daily intake of all 14 OUVFs through swimming was found to be 40-48 ng/kg/day. Ecological risk assessment indicates that BP-3 presents a medium risk for marine microalgae with a concurrent low risk for corals. The correlation analysis underscores a substantial interrelation of OUVFs in both coastal waters and beach deposits with various economic indicators, including annual rainfall, overnight tourists, total hotel rooms (unit), room occupancy rate, and sewage treatment capacity.

Occurrence, distribution, and risk assessment of antibiotics in a typical aquaculture area around the Dongzhai Harbor mangrove forest on Hainan Island.

The trees of the Dongzhai Harbor mangrove forest suffer from antibiotic contamination from surrounding aquaculture areas. Despite this being one of the largest mangrove forests in China, few studies have focused on the antibiotic pollution status in these aquaculture areas. In the present study, the occurrence, distribution, and risk assessment of 37 antibiotics in surface water and sediment samples from aquaculture areas around Dongzhai Harbor mangrove forests were analyzed. The concentration of total antibiotics (∑antibiotics) ranged from 78.4 ng/L to 225.6 ng/L in surface water (except S14-A2) and from 19.5 ng/g dry weight (dw) to 229 ng/g dw in sediment. In the sediment, the concentrations of ∑antibiotics were relatively low (19.5-52.3 ng/g dw) at 75 % of the sampling sites, while they were high (95.7-229.0 ng/g dw) at a few sampling sites (S13-A1, S13D, S8D). The correlation analysis results showed that the Kd values of the 9 antibiotics were significantly positively correlated with molecular weight (MW), Kow, and LogKow. Risk assessment revealed that sulfamethoxazole (SMX) in surface water and SMX, enoxacin (ENX), ciprofloxacin (CFX), enrofloxacin (EFX), ofloxacin (OFX), and norfloxacin (NFX) in sediment had medium/high risk quotients (RQs) at 62.5 % and 25-100 %, respectively, of the sampling sites. The antibiotic mixture in surface water (0.06-3.36) and sediment (0.43-309) posed a high risk at 37.5 % and 66.7 %, respectively, of the sampling sites. SMX was selected as an indicator of antibiotic pollution in surface water to assist regulatory authorities in monitoring and managing antibiotic pollution in the aquaculture zone of Dongzhai Harbor. Overall, the results of the present study provide a comprehensive and detailed analysis of the characteristics of antibiotics in the aquaculture environment around the Dongzhai Harbor mangrove system and provide a theoretical basis for the source control of antibiotics in mangrove systems.

Deciphering the disturbance mechanism of BaP on the symbiosis of Montipora digitata via 4D-Proteomics approach.

The coral holobiont is mainly composed of coral polyps, zooxanthellae, and coral symbiotic microorganisms, which form the basis of coral reef ecosystems. In recent years, the severe degradation of coral reefs caused by climate warming and environmental pollution has aroused widespread concern. Benzo(a)pyrene (BaP) is a widely distributed pollutant in the environment. However, the underlying mechanisms of coral symbiosis destruction due to the stress of BaP are not well understood. In this study, diaPASEF proteomics and 16S rRNA amplicon pyrosequencing technology were used to reveal the effects of 50 µg/L BaP on Montipora digitate. Data analysis was performed from the perspective of the main symbionts of M. digitata (coral polyps, zooxanthellae, and coral symbiotic microorganisms). The results showed that BaP impaired cellular antioxidant capacity by disrupting the GSH/GSSG cycle, and sustained stress causes severe impairment of energy metabolism and protein degradation in coral polyps. In zooxanthellae, BaP downregulated the protein expression of SOD2 and mtHSP70, which then resulted in oxidative free radical accumulation and apoptosis. For coral symbiotic microorganisms, BaP altered the community structure of microorganisms and decreased immunity. Coral symbiotic microorganisms adapted to the stress of BaP by adjusting energy metabolism and enhancing extracellular electron transfer. BaP adversely affected the three main symbionts of M. digitata via different mechanisms. Decreased antioxidant capacity is a common cause of damages to coral polyps and zooxanthellae, whereas coral symbiotic microorganisms are able to appropriately adapt to oxidative stress. This study assessed the effects of BaP on corals from a symbiotic perspective, which is more comprehensive and reliable. At the same time, data from the study supports new directions for coral research and coral reef protection.

Heterogeneous aggregation between microplastics and microalgae: May provide new insights for microplastics removal.

Existing studies have shown that microplastics (MPs) as artificial surfaces can be colonized by plankton microorganisms. However, systematic research on exploring the aggregation formation process of MPs and microalgae is still lacking and particularly the influencing factors of aggregation remain to be elucidated. Therefore, this study investigated the heterogeneous aggregation process between various microalgal species (i.e., Chlorella vulgaris, Scenedesmus obliquus, Tetraselmis subcordiformis, Chaetoceros müelleri and Streptococcus westermani) and MPs (i.e., mPS and mPLA) with different sizes (i.e., 74 µm and 613 µm), concentrations (i.e., 0.1 g/L, 1 g/L and 2 g/L) and shapes (i.e., the particle and sheet). The results showed that microalgae can first attach to the holes or protrusions of MPs and highly accumulate in the local region, and then multi-layer aggregation can be formed subsequently. The aggregation degree between MPs and microalgae was closely related to the MPs shape and size, and was less related to the MPs concentration. The aggregation speed of small-sized MPs (e.g., 74 µm) was faster than the large-sized ones (e.g., 613 µm). The MPs in a shape of sheet were more obvious than those in particle on their aggregation with microalgae. The density of aggregates was increased compared with pristine MPs, which is related to the cell density and cell number of attached microalgae. For the same type of MPs, the aggregation degree for the tested microalgae was as follows: Scenedesmus obliquus > C. vulgaris > T. subcordiformis > C. müelleri > S. westermani. Meanwhile, MPs inhibited cell growth of microalgae, particularly under the circumstance of their aggregation, by limiting the gas and mass transfer between microalgal cells and the extracellular environment. The heterogeneous aggregation of MPs and microalgae may provide new ideas for treatment and controlling of MPs in the environment.

Occurrence and Distribution of Antibiotics in a Tropical Mariculture Area of Hainan, China: Implications for Risk Assessment and Management.

With the rapid global demand for mariculture products in recent years, the use of antibiotics has increased intensively in the mariculture area. Current research on antibiotic residues in mariculture environments is limited, and less information is available on the presence of antibiotics in tropical waters, limiting a comprehensive understanding of their environmental presence and risk. Therefore, this study investigated the environmental occurrence and distribution of 50 antibiotics in the near-shore aquaculture waters of Fengjia Bay. A total of 21 antibiotics were detected in 12 sampling sites, including 11 quinolones, 5 sulfonamides, 4 tetracyclines, and 1 chloramphenicol; the quinolones pyrimethamine (PIP), delafloxacin (DAN), flurofloxacin (FLE), ciprofloxacin (CIP), norfloxacin (NOR), pefloxacin (PEF), enrofloxacin (ENO), and minocycline (MNO) of the tetracycline class were detected in all sampling points. The total antibiotic residue concentrations in the study area ranged from 153.6 to 1550.8 ng/L, the tetracycline antibiotics were detected in the range of 10 to 1344.7 ng/L, and the chloramphenicol antibiotics were detected in the range of 0 to 106.9 ng/L. The detected concentrations of quinolones ranged from 81.3 to 136.1 ng/L, and the residual concentrations of sulfonamide antibiotics ranged from 0 to 313.7 ng/L. The correlation analysis with environmental factors revealed that pH, temperature, conductivity, salinity, NH3--N, and total phosphorus had a strong correlation with antibiotics. Based on PCA analysis, the main sources of antibiotic pollution in the area were determined to be the discharge of farming wastewater and domestic sewage. The ecological risk assessment indicated that the residual antibiotics in the water environment of the near-shore waters of Fengjiawan had certain risks to the ecosystem. Among them, CIP, NOR, sulfamethoxazole (TMP), ofloxacin (OFL), enrofloxacin (ENO), sulfamethoxazole (SMX), and FLE showed medium to high risk. Therefore, it is recommended to regulate the use of these antibiotics and the discharge and treatment of culturing wastewater, and measures should be taken to reduce the environmental pollution caused by antibiotics and to monitor the long-term ecological risk of antibiotics in the region. Overall, our results provide an important reference for understanding the distribution and ecological risk of antibiotics in Fengjiawan.

An integrative investigation of developmental toxicities induced by triphenyltin in a larval coral reef fish, Amphiprion ocellaris.

Triphenyltin (TPT) is widely distributed on coastlines, which makes coral reef fish a potential target of TPT pollution. However, the negative effects of TPT on coral reef fish remain poorly understood. Therefore, in the present study, the larval coral reef fish Amphiprion ocellaris was used to investigate the developmental toxicities of TPT at environmentally relevant concentrations (0, 1, 10 and 100 ng/L). After TPT exposure for 14 d, the cumulative mortality increased, and growth was suppressed. In addition, TPT exposure inhibited the development of melanophores and xanthophores and delayed white strip formation, which might be responsible for the disruption of the genes (erbb3b, mitfa, kit, xdh, tyr, oca2, itk and trim33) related to pigmentation. TPT exposure also attenuated ossification of head skeletal elements and the vertebral column and inhibited the expression of genes (bmp2, bmp4 and sp7) related to skeletal development. The observed developmental toxicities on growth, pigmentation and skeleton development might be associated with the disruption of thyroid hormones and the genes related to thyroid hormone regulation (tshß, thrα, thrß, tg, tpo, dio2, and ttr). In addition, TPT exposure interfered with locomotor and shoaling behavior, and the related genes dbh, avp and avpr1aa. Taken together, our results suggest that TPT pollution might threaten the development of one of the most iconic coral reef fish, which might produce disastrous consequences on the health of coral reef ecosystems.

Environmental concentrations of benzophenone-3 disturbed lipid metabolism in the liver of clown anemonefish (Amphiprion ocellaris).

Benzophenone-3 (BP-3) often used as a UV filter in various products and an endocrine disruptor. In this work, we exposed the clown anemonefish to 10 µg/L and 50 µg/L BP-3 for 7 and 14 days. Liver histological, biochemical analysis, and transcriptome sequencing were used to explore the mechanism of the lipid metabolism disorder in the liver of three-month-old clown anemonefish treated with BP-3. The histological and biochemical analysis showed that BP-3 induces morphological changes and lipid droplet accumulation, and the lipid content, lipase, and antioxidant enzyme activity were abnormal. After treatment with 10 µg/L and 50 µg/L BP-3 for 7 days, the transcriptome analysis further demonstrated that the KEGG analysis revealed that the differentially expressed genes (DEGs) were mainly associated with fat digestion and absorption, PPAR signaling pathway, circadian rhythm, and mineral absorption pathways; After 10 µg/L and 50 µg/L of BP-3 exposure for 14 days, the KEGG analysis were mainly associated with circadian rhythm, circadian rhythm-fly, protein processing in the endoplasmic reticulum, and beta-alanine metabolism pathways. Several key genes were involved in the process of liver lipid metabolism, including CD36, APoA-Ⅰ, FABP, LPL, ACS, and PEPCK. The qRT-PCR validation results showed that eight genes (CYP8B1, FABP1, LPL, MGAT, PEPCK, PER1, PSMB4, PSME2) were significantly down-regulated, and the other two genes (Fbxl3, RXR) were significantly up-regulated after 7 days of BP-3 exposure. Similarly, eleven genes (AMPK, ARNTL, Bmal1, CASP3, CYC, CYP2J, CYP2U1, GSK3A, PEPCK, RAC1, RORA) were significantly up-regulated, and the other four genes (NR1D1, PER1, PTGDS, HLF) were significantly down-regulated after 14 days of BP-3 exposure. In conclusion, our results elucidate the physiological and molecular responses to BP-3 exposure in the liver lipid metabolism of clown anemonefish, and these findings reveal that the regulation of lipid metabolism is disturbed when clown anemonefish is exposed to UV filters.

Mechanistic toxicity and growth abnormalities mediated by subacute exposure to environmentally relevant levels of benzophenone-3 in clown anemonefish (Amphiprion ocellaris).

Benzophenone-3 (BP-3) is a UV filter that is ubiquitously present in the environment due to its photostability and degradation resistance and has wide applications in personal care products. BP-3 will eventually be discharged into the ocean. Studies shows BP-3 interferes with endocrine system of aquatic organisms, especially fish. However, the toxicity and mechanisms of subacute exposure of the coral reef fish to BP-3 remain elusive. Here, we exposed the one-month-old clown anemonefish to BP-3 at 1 and 10 µg/L for 14 and 28 days, respectively. After chronic exposure, the effects of BP-3 on the growth of clown anemonefish were investigated in terms of growth-related hormones, immune enzyme activity, digestive enzyme activity, transcriptional profiling of feeding- and obesity-related genes and digital RNA sequencing. The body weight in the BP-3 groups were abnormally increased (1 µg/L group in 14 days treatment and all groups in 28 days treatment), altered insulin content (28 days exposure), immune-related and digestive-related enzymatic activities. At the molecular level, BP-3 interferes with the expression of feeding- and obesity-related genes. Digital RNA sequencing analysis showed that BP-3 interferes with Kyoto encyclopedia of genes and genomes (KEGG) pathways related to growth, social behavior (learning behavior), Mitogen-Activated Protein Kinase (MAPK) signaling pathway, PI3K-Akt signaling pathway, and insulin secretion. Notably, in the insulin secretion, BP-3 induced Ca2+ up-regulation that may damage ß cells. Growth abnormalities and social behavior (learning behavior) KEGG pathway disturbances may have potential impacts on populations of clown anemonefish. Our results reveal the toxicological effects of subacute exposure to BP-3, and provides insight into the effects and mechanisms of BP-3 on clown anemonefish growth.

Genomic Evidence for the Recycling of Complex Organic Carbon by Novel Thermoplasmatota Clades in Deep-Sea Sediments.

Thermoplasmatota have been widely reported in a variety of ecosystems, but their distribution and ecological role in marine sediments are still elusive. Here, we obtained four draft genomes affiliated with the former RBG-16-68-12 clade, which is now considered a new order, "Candidatus Yaplasmales," of the Thermoplasmatota phylum in sediments from the South China Sea. The phylogenetic trees based on the 16S rRNA genes and draft genomes showed that "Ca. Yaplasmales" archaea are composed of three clades: A, B, and C. Among them, clades A and B are abundantly distributed (up to 10.86%) in the marine anoxic sediment layers (>10-cm depth) of six of eight cores from 1,200- to 3,400-m depths. Metabolic pathway reconstructions indicated that all clades of "Ca. Yaplasmales" have the capacity for alkane degradation by predicted alkyl-succinate synthase. Clade A of "Ca. Yaplasmales" might be mixotrophic microorganisms for the identification of the complete Wood-Ljungdahl pathway and putative genes involved in the degradation of aromatic and halogenated organic compounds. Clades B and C were likely heterotrophic, especially with the potential capacity of the spermidine/putrescine and aromatic compound degradation, as suggested by a significant negative correlation between the concentrations of aromatic compounds and the relative abundances of clade B. The sulfide-quinone oxidoreductase and pyrophosphate-energized membrane proton pump were encoded by all genomes of "Ca. Yaplasmales," serving as adaptive strategies for energy production. These findings suggest that "Ca. Yaplasmales" might synergistically transform benthic pollutant and detrital organic matter, possibly playing a vital role in the marine and terrestrial sedimentary carbon cycle. IMPORTANCE Deep oceans receive large amounts of complex organic carbon and anthropogenic pollutants. The deep-sea sediments of the continental slopes serve as the biggest carbon sink on Earth. Particulate organic carbons and detrital proteins accumulate in the sediment. The microbially mediated recycling of complex organic carbon is still largely unknown, which is an important question for carbon budget in global oceans and maintenance of the deep-sea ecosystem. In this study, we report the prevalence (up to 10.86% of the microbial community) of archaea from a novel order of Thermoplasmatota, "Ca. Yaplasmales," in six of eight cores from 1,200- to 3,400-m depths in the South China Sea. We provide genomic evidence of "Ca. Yaplasmales" in the anaerobic microbial degradation of alkanes, aliphatic and monoaromatic hydrocarbons, and halogenated organic compounds. Our study identifies the key archaeal players in anoxic marine sediments, which are probably critical in recycling the complex organic carbon in global oceans.

Physiological and muscle tissue responses in Litopenaeus vannamei under hypoxic stress via iTRAQ.

White L. vannamei have become the most widely cultivated shrimp species worldwide. Cultivation of L. vannamei is one of the predominant sectors in China's aquaculture industry. This study focused on the physiological and biochemical responses, differential protein expression, and expression characteristics of the related crucial functional protein genes under low oxygen conditions among different strains of L. vannamei. It was found that 6 h of hypoxic stress caused a significant reduction in the total hemocyte number in both strains, while the hypoxia-sensitive strain showed a stronger reduction. In contrast, the hemocyanin concentration showed only an overall upward trend. Proteomic analysis of L. vannamei muscle tissue revealed 3,417 differential proteins after 12 h of hypoxic stress. Among them, 29 differentially expressed proteins were downregulated and 244 were upregulated in the hypoxia-sensitive strain. In contrast, there were only 10 differentially expressed proteins with a downregulation pattern and 25 with an upregulation pattern in the hypoxia-tolerant strain. Five protein genes that responded significantly to hypoxic stress were selected for quantitative real-time PCR analysis, namely, hemocyanin, chitinase, heat shock protein 90 (HSP 90), programmed death protein, and glycogen phosphorylase. The results showed that the gene expression patterns were consistent with proteomic experimental data except for death protein and glycogen phosphorylase. These results can enrich the general knowledge of hypoxic stress in L. vannamei and the information provided differentially expressed proteins which may be used to assist breeding programs of L. vannamei of new strains with tolerance to hypoxia.