Phospholipid Compositions in Portunus trituberculatus Larvae at Different Developmental Stages

Phospholipids are used to improve the growth and survival of Portunus trituberculatus,a widely cultured crab species in China.However,only total phospholipids or several classes are applied in crab diets.In this study,we employed a targeted lipidomic method to investigate the comprehensive phospholipid composition in P.trituberculatus larvae and reveal the changing phospholipid profile over the larval development.Results showed that P.trituberculatus larvae contain 112 phospholipid species belonging to 10 phospholipid classes,in which phosphatidylcholine(PC)and phosphatidylethanolamine(PE)species are the most abundant,and PC,PE,phosphatidic acid,and phosphatidylserine(PS)are with high concentrations.The levels of all phospholipids significantly changed with larval development,which was highlighted by the downward parabolic changes in PE,phosphatidylglycerol,phosphatidylino-sitol,PS,lysophosphatidic acid,and sphingomyelin levels.In addition,nearly all phospholipid species were depleted at the M stage,which probably contributed to the mass mortality of crab larvae.These findings on the composition and alterations of phospholipids in P.trituberculatus larvae provide novel perspectives for the targeted supplementation of phospholipids in crab diets.Our work also highlights the use of targeted UHPLC-MS lipidomics in understanding the changes of phospholipids during crab development.     

南海海域三疣梭子蟹的幼体培育

幼体期Z1和Z2投喂扁藻和轮虫,Z2后期开始以轮虫和卤虫无节幼体混合投喂,Z3后用卤虫无节幼体投喂,大眼幼体用不同饵料培育,进行南海海域三疣梭子蟹幼体培育,结果表明:溞状幼体平均成活率为15.3%;Z2变态为Z3、Z4变态为M为溞状幼体培育的敏感期,幼体的阶段成活率较低;以卤虫无节幼体、冰冻卤虫成虫、活卤虫成虫为饵料,均能使培育大眼幼体至幼蟹,其中活卤虫成虫投喂、加防残网片培育效果最好,成活率达80%。     

Disruption of bacterial balance in the gut of Portunus trituberculatus induced by Vibrio alginolyticus infection

Journal of Oceanology and Limnology - Gut microbiota impacts the health of crustaceans. Vibrio alginolyticus is a main causative pathogen that induces the vibriosis in farmed swimming crabs,...     

IgE Reactivity of Potential Allergens from the Swimming Crab Portunus trituberculatus and the Research of ELISA Reagent for Detecting Crab Food Anaphylaxis

Crustacean is one of the major allergic foods.It is of great significance to identify more crab allergens and research the detection methods for crab food anaphylaxis.In this study,IgE reactivity to three recombinant proteins from Portunus trituberculatus,including tropomyosin(rPtTM),myosin light chain(rPtMLC),and pancreatic lipase(rPtPL),were detected by enzyme-linked im-munosorbent assay(ELISA).The expressions of tropomyosin(TM)in various tissues of P.trituberculatus were detected by Western blot(WB).Furthermore,microplates were coated with rPtTM and the ELISA conditions were optimized.The cut-off value was deter-mined by the receiver operating characteristic(ROC)curve.Among 51 crab-allergic sera,21(41.2%)showed positive IgE to rPtTM.Other 70 crab-allergic sera more frequently recognized rPtPL(9/70;12.9%),followed by rPtMLC(1/70;1.4%).WB results showed that TM was mainly expressed in the muscle,followed by the heart and a small amount in gills and the testis.The optimal results showed that the coating condition of rPtTM was 50 ng per well with coating for 3 h at 37℃.The optimal blocking condition was 1.2%BSA with blocking for 3 h at 37℃,and the optimal dilution of the second antibody was 1:1500.The ROC curve showed that the ELISA reagent had high sensitivity(83.52%)and specificity(98.00%)when the cut-off value was 0.45.All results indicated that tropomyosin is the major allergen of P.trituberculatus,and myosin light chain and pancreatic lipase are the potential allergens.Addi-tionally,the ELISA reagent developed with the rPtTM was feasible for laboratory detection of crab anaphylaxis.     

Polymorphisms of clip domain serine proteinase and serine proteinase homolog in the swimming crab Portunus trituberculatus and their association with Vibrio alginolyticus

Clip domain serine proteases(cSPs) and their homologs(SPHs) play an important role in various biological processes that are essential components of extracellular signaling cascades,especially in the innate immune responses of invertebrates.Here,polymorphisms of PtcSP and PtSPH from the swimming crab Portunus trituberculatus were investigated to explore their association with resistance/susceptibility to Vibrio alginolyticus.Polymorphic loci were identified using Clustal X,and characterized with SPSS 16.0 software,and then the significance of genotype and allele frequencies between resistant and susceptible stocks was determined by a χ~2 test.A total of 109 and 77 single nucleotide polymorphisms(SNPs) were identified in the genomic fragments of PtcSP and PtSPH,respectively.Notably,nearly half of PtSPH polymorphisms were found in the non-coding exon 1.Fourteen SNPs investigated were significantly associated with susceptibility/resistance to V.alginolyticus(P0.05).Among them,eight SNPs were observed in introns,and one synonymous,four non-synonymous SNPs and one ins-del were found in coding exons.In addition,five simple sequence repeats(SSRs) were detected in intron 3 of PtcSP.Although there was no statistically significant difference of allele frequencies,the SSRs showed different polymorphic alleles on the basis of the repeat number between resistant and susceptible stocks.After further validation,polymorphisms investigated here might be applied to select potential molecular markers of P.trituberculatus with resistance to V.alginolyticus.     

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly inc...     

墨西哥湾扇贝稚贝盐度适应性的研究

在室内控制条件下,观察不同海水盐度对墨西哥湾扇贝(Argopecten irradians concentricusSay)稚贝存活及生长的影响,并将168 h 50%死亡的盐度作为适宜生存临界盐度,将增长率为最佳增长率30%时所对应的盐度作为适宜生长临界盐度。结果表明:在稚贝壳高4.38 mm±0.31 mm,水温25.4~27.8℃,pH值7.8~8.2下,墨西哥湾扇贝稚贝适宜生存盐度为23.6~35.7,最适生存盐度为26.0~31.3;适宜生长盐度为23.9~35.8,最适生长盐度为28.7~31.3。在盐度变化超出最适范围时,`墨西哥湾扇贝稚贝对低盐更为敏感,高盐比低盐对稚贝存活、生长、增重的影响小。当盐度为28.7时,墨西哥湾扇贝稚贝具有最高的相对存活率(100%)、壳高日增长率(0.116 2 mm.d-1)及体重日增长率(1.95 mg.d-1)。当盐度在最适范围之外升高或降低,稚贝的存活率、壳高及体重日增长率也随着降低。     

A New Method to Predict Erythrocyte Sedimentation Rate with Natural Geographical Factors and Location by Case-based Reasoning: A Case Study of China

Reference values of erythrocyte sedimentation rate(ESR)are the key to interpret ESR blood test in clinic.The common local reference ESR values are more accuracy in blood test that are established with natural geographical factors by using the multiple linear regression(MLR)model and the artificial neural network(ANN).These knowledge-based methods have limitations since the knowledge domains of ESR and natural geographical factors are limited.This paper presents a new cases-depended model to establish reference ESR values with natural geographical factors and location using case-based reasoning(CBR)since knowledge domain of ESR and geographical factors is weak.Overall 224 local normal ESR values of China that calculated from 13623 samples were obtained,and the corresponding natural geographical factors and location that include altitude,sunshine hours,relative humidity,temperature,precipitation,annual temperature range and annual average wind speed were obtained from the National Geomatics Center of China.CBR was used to predict the unseen local reference ESR values with cases.The average absolute deviation(AAD),mean square error(MSE),prediction accuracy(PA),and Pearson correlation coefficient(r)between the observed and estimated data of proposed model is 33.07%,9.02,66.93% and 0.78,which are better than those of ANN and MLR model.The results show that the proposed model provides higher prediction accuracy than those of the artificial neural network and multiple linear regression models.The predicted values are very close to the observed values.Model results show significant agreement of cases data.Consequently,the model is used to predict the unseen local reference ESR with natural geographical factors and location.In spatial,the highest ESR reference areas are distributed in the southern-western district of China that includes Sichuan,Chongqing,Guangxi and Guizhou provinces,and the reference ESR values are greater than 23 mm/60 min.The higher ESR reference values are distributed in the middle part and northern-eastern of China which include Hubei,Henan,Shaanxi,Shanxi,Jilin and Heilongjiang provinces,and the reference ESR values are greater than 18 mm/60min.The lowest ESR reference values are distributed in the northern-western of China that includes Tibet and Xinjiang,and the reference ESR values are lower than 5 mm/60min.