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Ding Y.,Xiamen University | Ding Y.,State Oceanic Administration | Ai C.,Xiamen University | Mu Y.,State Oceanic Administration | And 5 more authors.
Fish and Shellfish Immunology | Year: 2016

Interleukin-17s (IL-17s) play critical roles in inflammatory response and host defense against extracellular pathogens. IL-17s induce the immune response signaling through the specific IL-17 receptors (IL-17Rs) that consist of five members (IL-17RA to E). In the present work, we have identified the five IL-17R orthologs (LycIL-17Rs) from large yellow croaker Larimichthys crocea. The deduced protein of each LycIL-17R exhibits a typical IL-17R domain architecture, including a signal peptide, the extracellular FNIII domain (IL-17RA/RB/RD) or IL-17_R_N domain (IL-17RC/RE), a transmembrane domain, and a SEFIR domain in cytoplasmic region. In particular, the extracellular regions of teleost IL-17RB are much shorter than those in mammals and lack an FNIII domain (FN2). Phylogenetic tree shows that IL-17Rs are classified into two main groups: IL-17RA/RB/RD group and IL-17RC/RE group, which is distinct from previous proposal that grouped IL-17RB into IL-17RC/RE. The surrounding genes of IL-17Rs are conservatively aligned in genomes between teleosts and mammals. The five LycIL-17Rs were constitutively expressed in all tissues examined, but with different expression patterns. Aeromonas hydrophila infection significantly upregulated LycIL-17RA, RC, RD and RE in both mucosal tissue (gills) and systemic immune tissues (head kidney and spleen), while the increase of LycIL-17RB expression could be detected in gills, indicating that LycIL-17Rs may be involved in host defense against bacterial infection. Thus, these results suggest that teleost IL-17Rs may function in mediating immune response as their mammalian orthologs. To our knowledge, this is the first report of molecular characterization of the five IL-17Rs (IL-17RA/RB/RD and IL-17RC/RE) in teleost fish. © 2016 Elsevier Ltd

Li M.,State Oceanic Administration | Li M.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | Guo W.,State Oceanic Administration | Guo W.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | And 3 more authors.
Applied Microbiology and Biotechnology | Year: 2016

The reductase component (MhpP) of the Sulfobacillus acidophilus TPY multicomponent phenol hydroxylase exhibits only 40 % similarity to Pseudomonas sp. strain CF600 phenol hydroxylase reductase. Amino acid sequence alignment analysis revealed that four cysteine residues (Cys-X4-Cys-X2-Cys-X29-35-Cys) are conserved in the N terminus of MhpP for [2Fe-2S] cluster binding, and two other motifs (RXYS and GXXS/T) are conserved in the C terminus for binding the isoalloxazine and phosphate groups of flavin adenine dinucleotide (FAD). Two motifs (S/T-R and yXCGp) responsible for binding to reduce nicotinamide adenine dinucleotide phosphate (NADPH) are also conserved in MhpP, although some residues differ. To confirm the function of this reductase, MhpP was heterologously expressed in Escherichia coli BL21(DE3) and purified. UV-visible spectroscopy and electron paramagnetic resonance spectroscopy revealed that MhpP contains a [2Fe-2S] cluster. MhpP mutants in which the four cysteine residues were substituted via site-directed mutagenesis lost the ability to bind the [2Fe-2S] cluster, resulting in a decrease in enzyme-specific oxidation of NADPH. Thin-layer chromatography revealed that MhpP contains FAD. Substrate specificity analyses confirmed that MhpP uses NADPH rather than NADH as an electron donor. MhpP oxidizes NADPH using cytochrome c, potassium ferricyanide, or nitro blue tetrazolium as an electron acceptor, with a specific activity of 1.7 ± 0.36, 0.78 ± 0.13, and 0.16 ± 0.06 U/mg, respectively. Thus, S. acidophilus TPY MhpP is a novel NADPH-dependent reductase component of phenol hydroxylase that utilizes FAD and a [2Fe-2S] cluster as cofactors. © 2016 Springer-Verlag Berlin Heidelberg

Guo X.,Xiamen University | Guo X.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | Xu X.,Xiamen University | Xu X.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | And 7 more authors.
Journal of Experimental Marine Biology and Ecology | Year: 2016

Increasing atmospheric CO2 can decrease the seawater pH and carbonate ions, which may adversely affect the larval survival of calcareous animals. In this study, we simulated future atmospheric CO2 concentrations (800, 1500, 2000 and 3000 μatm) and examined the effects of ocean acidification on the embryonic and larval stage of an infaunal clam Paphia undulate. Significant decrease of hatching of P. undulate was observed when the pCO2 reached 3000 μatm, and larval deformation rate increased significantly when pCO2 reached 2000 μatm, indicating a strong tolerance to ocean acidification compared with the embryonic development of other bivalves. The larvae cultured in 1500 μatm pCO2 exhibited the fastest growth, highest survival and shortened planktonic period, which unordinary phenomenon reflected the beneficial effect of ocean acidification on P. undulate larval development. The better development of P. undulate larvae under a higher CO2 condition maybe an adaptation in response to the acidified sediment in which they live. © 2016 Elsevier B.V.

Bai J.,Xiamen University | Gong W.,Xiamen University | Wang C.,Xiamen University | Gao Y.,Xiamen University | And 4 more authors.
Fish Physiology and Biochemistry | Year: 2015

In vertebrates, the aromatase coded by the cyp19a1a gene can catalyze the conversion from androgens to estrogens. Thus, the regulatory mechanisms of cyp19a1a gene expression are a critical research field in reproductive endocrinology. In this study, we use zebrafish as a model to study the dynamic methylation levels of the cyp19a1a gene core promoter during zebrafish ovarian folliculogenesis. The results show that there is an apparent fluctuation of the methylation levels of zebrafish cyp19a1a core promoter. Moreover, the methylation levels are inversely correlated with the expression levels of cyp19a1a transcripts when the ovarian follicles develop from PV into the MV stage. Also, the CpG dinucleotides which are close to the transcriptional starting site may have provided a significant blocking effect on inhibiting the transcriptional function of RNA polymerase II. Taken together, the results from the present study strongly suggest that DNA methylation was one of mechanisms that are involved in the regulation of cyp19a1a gene expression during folliculogenesis. This methylation mechanism modifying transcriptional process accompanied with zebrafish ovarian folliculogenesis might also shed new light on the regulation of cyp19a1a expression during the ovarian developmental stage in other vertebrates. © 2015 Springer Science+Business Media Dordrecht

Shu L.,Xiamen University | Yang Y.,Xiamen University | Huang H.,Xiamen University | Ye H.,Xiamen University | Ye H.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources
Molecular and Cellular Endocrinology | Year: 2016

In vertebrates, bone morphogenetic proteins (BMPs) play an important role in various biological processes. However, the function of BMPs in crustaceans is still unknown. In our study, a ligand (BMP7) and two receptors (Sp-BMPRIB and Sp-BMPRII) are cloned firstly in the mud crab, Scylla paramamosain. The qRT-PCR demonstrated that both ligand and receptors were expressed in various tissues, especially in ovary. The expression of BMPRs mRNA increased along the ovarian development, while BMP7 had an opposite tendency. In-situ hybridization revealed that Sp-BMPRIB and Sp-BMPRII were expressed in both oocytes and follicle cells, whereas Sp-BMP7 was exclusively localized in follicle cells. RNAi experiments showed that the expression levels of Smad1 and vitellogenin receptor declined rapidly after BMPRs were silenced. Based on these data, we hypothesized that in S. paramamosain, BMP7 and BMPRs had impact on the ovarian development, presumably via the autocrine/paracrine way. © 2016 Elsevier Ireland Ltd

Lin Z.,Jimei University | Lin Z.,Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering | Wang D.,Jimei University | Peng A.,Jimei University | And 4 more authors.
Journal of Separation Science | Year: 2016

A selective sample cleanup method using molecularly imprinted polymers was developed for the separation of domoic acid (a shellfish toxin) from shellfish samples. The molecularly imprinted polymers for domoic acid was prepared by emulsion polymerization using 1,3,5-pentanetricarboxylic acid as the template molecule, 4-vinyl pyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinker, and Span80/Tween-80 (1:1 v/v) as the composite emulsifiers. The molecularly imprinted polymer showed high affinity to domoic acid with a dissociation constant of 13.5 μg/mL and apparent maximum adsorption capacity of 1249 μg/g. They were used as a selective sorbent for the detection of domoic acid from seafood samples coupled with high-performance liquid chromatography. The detection limit of 0.17 μg/g was lower than the maximum level permitted by several authorities. The mean recoveries of domoic acid from clam samples were 93.0–98.7%. It was demonstrated that the proposed method could be applied to the determination of domoic acid from shellfish samples. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Li L.,Jimei University | Lin Z.-Z.,Jimei University | Peng A.-H.,Jimei University | Zhong H.-P.,Jimei University | And 3 more authors.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2016

A direct competitive enzyme-linked immunosorbent assay (ELISA) method was used for the detection of malachite green (MG) with a high sensitivity and selectivity using magnetic molecularly imprinted polymers (MMIPs) as a bionic antibody. MMIPs were prepared through emulsion polymerization using Fe3O4 nanoparticles as magnetic nuclei, MG as a template, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent and span-80/tween-80 as mixed emulsifiers. The MMIPs were characterized by scanning electron micrographs (SEM), thermal-gravimetric analyzer (TGA), Fourier transform infrared spectrometer (FT-IR) and vibrating sample magnetometer (VSM), respectively. A high magnetic saturation value of 54.1 emu g−1 was obtained, resulting in rapid magnetic separation of MMIPs with an external magnet. The IC50 of the established ELISA method was 20.1 μg L−1 and the detection limit (based on IC85) was 0.1 μg L−1. The MMIPs exhibited high selective binding capacity for MG with cross-reactivities less than 3.9% for MG structural analogues. The MG spiking recoveries were 85.0%–106% with the relative standard deviations less than 4.7%. The results showed that the biomimetic ELISA method by using MMIPs as bionic antibody could be used to detect MG rapidly in fish samples with a high sensitivity and accuracy. © 2016 Elsevier B.V.

Luo Z.-H.,State Oceanic Administration | Luo Z.-H.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | Xu W.,State Oceanic Administration | Xu W.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | And 4 more authors.
Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology | Year: 2015

Nitrification, the aerobic oxidation of ammonia to nitrate via nitrite, is performed by nitrifying microbes including ammonia-oxidizing bacteria (AOB) and archaea (AOA). In the current study, the phylogenetic diversity and abundance of AOB and AOA in deep-sea sediments of the Pacific Ocean were investigated using ammonia monooxygenase subunit A (amoA) coding genes as molecular markers. The study uncovered 3 AOB unique operational taxonomic units (OTUs, defined at sequence groups that differ by ≤5 %), which indicates lower diversity than AOA (13 OTUs obtained). All AOB amoA gene sequences were phylogenetically related to amoA sequences similar to those found in marine Nitrosospira species, and all AOA amoA gene sequences were affiliated with the marine sediment clade. Quantitative PCR revealed similar archaeal amoA gene abundances [1.68 × 105–1.89 × 106 copies/g sediment (wet weight)] among different sites. Bacterial amoA gene abundances ranged from 5.28 × 103 to 2.29 × 106 copies/g sediment (wet weight). The AOA/AOB amoA gene abundance ratios ranged from 0.012 to 162 and were negatively correlated with total C and C/N ratio. These results suggest that organic loading may be a key factor regulating the relative abundance of AOA and AOB in deep-sea environments of the Pacific Ocean. © Springer International Publishing Switzerland 2015.

Gao C.,XingHuaLing District Food and Drug Administration | Jin M.,Third Institute of Oceanography | Yi Z.,Third Institute of Oceanography | Zeng R.,Third Institute of Oceanography | And 2 more authors.
Journal of Microbiology and Biotechnology | Year: 2015

A novel sulfatase gene, ary423 (1,536 bp ORF), encoding a protein of 511 amino acids with a calculated molecular mass of 56 kDa, was identified from Flammeovirga pacifica, which was isolated from deep-sea sediments of west Pacific Ocean. Amino acid sequence analysis revealed that Ary423 possessed a conserved C-X-A-X-R motif, which was recognized as the sulfatase signature. Phylogenetic analysis suggested that Ary423 belonged to arylsulfatases. After heterologous expression in Escherichia coli cells, the recombinant Ary423 was purified with a Ni+ affinity column, and was shown to be highly active at a broad range of temperatures from 30° to 70°C, with maximum activity at 40°C. Furthermore, recombinant Ary423 retained more than 70% and 40% of its maximum activity after 12 h of incubation at 50°C and 60°C, respectively, exhibiting good thermostability at high temperatures. The optimal pH for Ary423 was determined to be 8.0 and the activity of Ary423 could be slightly enhanced by Mg2+. The recombinant enzyme could hydrolyze sulfate ester bonds in p-nitrophenyl sulfate (NPS) and Asparagus crude polysaccharides with a specific activity of 64.8 U/mg and 25.4 U/mg, respectively. These favorable properties could make Ary423 attractive for application in the desulfating process of agar production. © 2015 by The Korean Society for Microbiology and Biotechnology.

Zhang Y.,State Oceanic Administration | Zhang Y.,Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources | Chen D.,China National Center for Food Safety Risk Assessment | Hong Z.,State Oceanic Administration | Hong Z.,China National Center for Food Safety Risk Assessment
Toxins | Year: 2015

In this study, we developed a self-assembly pipette tip solid-phase extraction (PTSPE) method using a high molecular weight polymer material (PAX) as the adsorbent for the determination of domoic acid (DA) in human urine samples by liquid chromatography high-resolution mass spectrometry (LC-HRMS) analysis. The PTSPE cartridge, assembled by packing 9.1 mg of PAX as sorbent into a 200 μL pipette tip, showed high adsorption capacity for DA owing to the strong cationic properties of PAX. Compared with conventional SPE, the PTSPE is simple and fast, and shows some advantages in the aspects of less solvent consumption, low cost, the absence of the evaporation step, and short time requirement. All the parameters influencing the extraction efficiency such as pH, the amount of sorbent, the number of aspirating/dispensing cycles, and the type and volume of eluent in PTSPE were carefully investigated and optimized. Under the optimized conditions, the limit of detection (LOD) and limit of quantification (LOQ) values of DA were 0.12 μg/L and 0.37 μg/L respectively. The extraction recoveries of DA from the urine samples spiked at four different concentrations were in a range from 88.4% to 102.5%. The intra- and inter-day precisions varied from 2.1% to 7.6% and from 2.6% to 12.7%, respectively. The accuracy ranged from -1.9% to -7.4%. © 2015 by the authors; licensee MDPI, Basel, Switzerland.

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