Establishing environmental background values of selected trace elements and environmental quality assessment of fi negrained sediments in the Jiaozhou Bay catchment,China

With the development of economy,the impact of human activities on ecological environment is increasing,and environmental protection work is important.Trace elements(Co,Ni,Cu,Pb,Zn,Cd,Cr,and Sc)in surface fi ne-grained sediment samples from the Jiaozhou Bay catchment were selected to evaluate their 2015 environmental background values and the environmental quality.Using statistical analysis,the environmental background values(ranges)of Sc,Co,Ni,Cu,Pb,Zn,Cd,and Cr were calculated,being 13.1(10.8-15.4),12.4(8.6-16.2),32.0(22.9-41.2),29.6(13.5-64.9),24.1(13.0-44.6),77.6(38.5-156.5),0.07(0.02-0.20),and 82.5(66.5-104.0)mg/kg,respectively.The enrichment factor,contamination factor,and pollution load index were used to evaluate the pollution status of the Jiaozhou Bay catchment.The environmental background values of most elements are higher than those of the upper continental crust and lower than those of global shale.At present,the pollution in the eastern Jiaozhou Bay is much higher than that in the western part.The results shall be helpful for future management for trace element pollution monitoring in the Jiaozhou Bay catchment.

Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers Plectropomus leopardus

Leopard coral groupers(Plectropomus leopardus),commercially bred in South China,are a significant economical fish species.In this study,by means of quantitative real-time PCR(qRT-PCR)technology,we assessed the stability of six common reference genes expression and selected the appropriate reference genes in leopard coral groupers with or without Vibrio harveyi stimulation at different time points.These data produced by qRT-PCR was handled via the geNorm,NormFinder,and BestKeeper software.The results revealed all the examined reference genes had manifest tissue-specific expression in different tissues.Prior to V.harveyi stimulation,RPL13 gene was the appropriate reference gene among eleven tissue types(blood,spleen,hepatopancreas,kidney,stomach,gill,heart,skin,muscle,intestine,brain)in leopard coral groupers.Under V.harveyi stimulation,the most reliable reference genes varied from tissue to tissue and were closely hinged upon different time points.At 6-h post-bacterial injection,the appropriate reference genes in hepatopancreas,spleen,kidney,and gill were Actin,B2M,UBCE,and Actin,respectively.At 9-and 12-h post-bacterial injection,the appropriate reference genes in hepatopancreas,spleen,kidney,and gill were RPL13,Actin,Actin,and Actin,respectively.If one reference gene was preferable,RPL13,Actin,Actin,and Actin could be selected as the reference gene in hepatopancreas,spleen,kidney,and gill of leopard coral groupers after V.harveyi infection,respectively.Expression profiles of two target genes(IL-6 and NK-lysin)were used to further validate reliability of these selected appropriate candidates.This study will lay a solid foundation for the future research on qRTPCR analysis of gene expression in leopard coral groupers.

Metabolomic and proteomic responses of Phaeodactylum tricornutum to hypoxia

Diatoms are important contributors to global net primary productivity,and play a crucial role in the biogeochemical cycles of carbon,phosphorus,nitrogen,iron,and silicon.Currently in some regions in the ocean,there’s a trend that carbon content is high while oxygen concentration is low,and the underlying mechanisms of diatoms’response to low oxygen environments are worth investigating.Phaeodactylum tricornutum is a model diatom whose genome has been sequenced;it provides a universal molecular toolbox and a stable transgenic expression system.Therefore,the study of the responses of P.tricornutum to low oxygen has not only fundamental research significance but also important ecologic al significance.In this study,growth rates were determined and proteomic analysis and metabolomic analysis were performed to examine P.tricornutum responses under different oxygen concentrations(2%oxygen concentration for hypoxic condition and 21%oxygen concentration for the normal condition(sterilized air)).Results show that the hypoxic environment inhibited the growth of P.tricornutum.In the hypoxic conditions,P.tricornutum could reset its metabolism pathways,including enhancement in lipid utilization,replenishment of tricarboxylic acid(TCA)cycle through the glyoxylic acid cycle,and down-regulation of photorespiration to reduce energy waste.Additionally,the stress resistance mechanism was activated to facilitate the adaptation to low oxygen conditions.This study helps to reveal the different metabolic changes to hypoxia of diatom from that of higher plants,which might be ascribed to their different habitats and needs further exploration in the future.

Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis

Humpback grouper Cromileptes altivelis is one commercial fish with considerable economic value.To determine the expression stabilities of six commonly used internal reference genes in C.altivelis challenged by Vibrio harveyi and viral nervous necrosis virus(VNNV)through quantitative real-time PCR(qRT-PCR),the expression levels of selected genes in five immune organs stimulated with pathogenic infection were carefully evaluated using algorithms of geNorm,NormFinder,and BestKeeper.The results show that the expre ssion stabilities of the six candidate inte rnal reference genes were diffe re nt.Under no rmal physiological conditions,RPL13 were identified as the most stably expressed genes among five different immune organs(liver,spleen,kidney,intestine,and gill).After V.harveyi stimulation,RPL13,RPL13,EF1 A,RPL13,and EF1 A were identified by geNorm,NormFinder,and BestKeeper as the most stable genes in liver,spleen,kidney,intestine,and gill,respectively.Combining these three algorithms suggested that under stimulation of VNNV,RPL13,EF1 A,Actin,RPL13,and Actin were as the most stable genes in liver,spleen,kidney,intestine,and gill,respectively.These results suggest that specific experiment conditions and tissue types shall be considered when selecting the reference genes in qRT-PCR analysis.This study provided a solid foundation for future studies on gene expression of C.altivelis under different conditions.

Involvement of the ammonium assimilation mediated by glutamate dehydrogenase in response to heat stress in the scleractinian coral Pocillopora damicornis

Glutamate dehydrogenase(GDH)plays an important role in the ammonium assimilation and nitrogen metabolism by catalyzing the reversible oxidative deamination of L-glutamate toα-ketoglutarate.In the present study,the potential functions of GDH in response to heat stre ss were explored in the scleractinian coral Pocillopora damicornis(designated as PdGDH).The cDNA of PdGDH contained an open reading frame of 1611 bp encoding a polypeptide of 536 amino acids,which exhibited the highest sequence identity to GDH of Stylophora pistillata(96%identity),and the deduced PdGDH protein was predicted to contain one GdhA domain(from Val95 to Tyr525).The recombinant protein of PdGDH(rPdGDH)was expressed in Escherichia coli BL21(DE3)-Transetta,and its catalytic activity was measured under different temperatures,pH conditions and epigallocatechin-3-gallate(EGCG,a GDH inhibitor)concentrations.The purified rPdGDH only used reduced coenzyme nicotinamide adenine dinucleotide(NADH)as coenzyme,and its highe st activity was observed at 35℃and pH 7.5,re spectively.The rPdGDH activity was negatively correlated with the concentration of EGCG,and was inhibited by more than half(65%,P<0.05)at 10mol/L EGCG.No significant alteration of PdGDH mRNA expression was detected at 12 h after exposure to heat and ammonium(P>0.05).Furthermore,the activities of NADH-GDH in the scleractinian coral P.damicornis increased significantly at 12 h after the heat and ammonium stress,and the NADH-GDH activity in the heat stress group(32.66 U/mg,P<0.05)was significantly higher than that in the heat and ammonium stress group(11.26 U/mg).These results collectively suggested that PdGDH,as a homologue of glutamate dehydrogenase in the scleractinian coral P.damicornis,could respond to heat stress at the protein level,which would have ability to further promote ammonium assimilation to increase the heat acclimatization of the coral-Symbiodiniaceae symbiotic association.

New insights into the influence of myo-inositol on carbohydrate metabolism during osmoregulation in Nile tilapia (Oreochromis niloticus)

A two-factor(23)orthogonal testwas conducted to investigate the effects of dietary myo-inositol(MI)on the osmoregulation and carbohydrate metabolism of euryhaline fish tilapia(Oreochromis niloticus)under sustained hypertonic stress(20 practical salinity units[psu]).6 diets containing either normal carbohydrate(NC,30%)or high carbohydrate(HC,45%)levels,with 3 levels(0,400 and 1,200 mg/kg diet)of MI,respectively,were fed to 540 fish under 20 psu for 8 weeks.Dietary MI supplementation significantly improved growth performance and crude protein content of whole fish,and decreased the content of crude lipid of whole fish(P<0.05).Curled,disordered gill lamella and cracked gill filament cartilage were observed in the gill of fish fed diets without MI supplementation.The ion transport capacity in gill was significantly improved in the 1,200 mg/kg MI supplementation groups compared with the 0 mg/kg MI groups(P<0.05).Moreover,the contents of Na^(+),K^(+),Cl^(-)in serum weremarkedly reduced with the dietary MI supplementation(P<0.05).The fish fed 1,200 mg/kg MI supplementation had the highest MI content in the gills and the lowest MI content in the serum(P<0.05).Additionally,the fish fed with 1,200 mg/kg MI supplementation had the highest MI synthesis capacity in gills and brain(P<0.05).Dietary MI markedly promoted the ability of carbohydrate metabolism in liver(P<0.05).Moreover,fish in the 1,200 mg/kg MI groups had the highest antioxidant capacity(P<0.05).This study indicated that high dietary carbohydrate would intensify stress,and impair the ability of osmoregulation in tilapia under a long-term hypersaline exposure.The supplementation of MI at 1,200 mg/kg in the high carbohydrate diet could promote carbohydrate utilization and improve the osmoregulation capacity of tilapia under long-term hypertonic stress.