Effects of the Dietary Probiotic Clostridium butyricum on Intestine Digestive and Metabolic Capacities, SCFA Content and Body Composition in Marsupenaeus japonicus

A 56-day feeding trial was performed to evaluate the effects of dietary probiotic Clostridium butyricum(CB) on intestine digestive and metabolic capacities, intestine short-chain fatty acids(SCFA) content and body composition of kuruma shrimp Marsupenaeus japonicus. Shrimp s were randomly allocated into 9 tanks, 30 each, and f ed with diets containing different levels of C. butyricum(1×10~9 cfug^(-1)): 0 mgg^(-1) feed(Control), 100 mgg^(-1) feed(CB-100), 200 mgg^(-1) feed(CB-200), while each level was triplicated. The results indicated that compared with the control group, the intestine pepsin(Pep) activity and 5-hydroxytryptamine(5-HT) concentration of two C. butyricum group s were both increased. Amylase(AM Y) and lipase(LPS) activities wer e only induced in CB-200 group. Alanine aminotransferase(ALT) and aspartate aminotransferase(A ST) activities of two C. butyricum group s showed no significant change. The α-amylase(AM Y) gene expression was induced in CB-200 group, and tryp sin gene expression of two C. butyricum treated group s were both induced. Intestine SCFA content and body composition analysis showed that the contents of propionic acid, butyric acid and the crude protein of two C. butyricum group s were all higher than those of control. These results revealed that C. butyricum can modulate intestine digestive and metabolic capacities, improve intestine SCFA content and body crude protein content in M. japonicus.

Molecular characteristics of three thymosin-repeat proteins from Marsupenaeus japonicus and their responses to WSSV infection

β-thymosins, a family of highly conserved peptides, play a vital role in wound-healing, angiogenesis,antimicrobial process and antiviral immunity. Three novel β-thymosin-repeat proteins, named mjthm4, mjthm3 and mjthm2, were cloned from Marsupenaeus japonicus using expressed sequence tags（EST） from suppression subtractive hybridization. The mjthm4, mjthm3 and mjthm2 c DNAs possessed open reading frames that encoded166, 128 and 90 amino acid residue polypeptides and contained four, three and two β-thymosin actin binding modules, respectively. Blast analysis demonstrated that mjthm4, mjthm3 and mjthm2 shared high homology with known invertebrate multi-repeat β-thymosins. These proteins are ubiquitously expressed in all of the examined tissues, and the transcriptional levels were highest in the intestine. Further investigation revealed that mjthm4,mjthm3 and mjthm2 were remarkably up-regulated 6 h after WSSV infection. Moreover, while mjthm4 transcriptional levels displayed no changes, mjthm3 and mjthm2 levels decreased in the virus-resistant shrimps.The results indicate that mjthm4, mjthm3 and mjthm2 are novel multi-repeat β-thymosin homologues, have a close relationship with WSSV infection, and might contribute to a better understanding of host defense and/or virus invasion interactions in shrimps.

Molecular characterization and expression analysis of a novel dual-CRD C-type lectin in kuruma shrimp(Marsupenaeus japonicus)

C-type lectins are among the most significant pattern recognition receptors（PRRs） found in invertebrate. They are a class of carbohydrate-binding proteins that can recognize specific sugar moieties on the surface of pathogens. In the present study, a novel C-type lecitn（termed Mj Lectin） from kuruma shrimp Marsupenaeus japonicus was identified. The full-length c DNA of Mj Lectin was 1 245 bp with a 1 011 bp open reading frame（ORF） that encoded a polypeptide of 336 amino acid residues. Mj Lectin consisted of two tandemly arrayed carbohydrate-recognition domains（CRDs）, unlike other reported M. japonicus C-type lectins with only one CRD. It showed a high similarity to other shrimp dual-CRD lectins. Among the Ca2＋-binding Site 2, the tripeptide motif dictating the carbohydrate binding specificity was exhibited as a rare mutant LPN（Leu134-Pro135-Asn136） in CRD1 and a traditional EPN（Glu299-Pro300-Asn301） in CRD2, respectively. Mj Lectin showed a specific expression pattern in both tissue and cellular levels, for its m RNA transcript was mainly expressed in the F-cells of the hepatopancreas. After white spot syndrome virus（WSSV） challenge（3.6×108 virions/μL）, the expression of Mj Lectin in the hepatopancreas was up-regulated significantly at 48 h（P〈0.01） compared with the control group. These results suggested that Mj Lectin might be involved in the innate immune defense against WSSV infection.

Genome-wide mining, characterization, and development of microsatellite markers in Marsupenaeus japonicus by genome survey sequencing

The kuruma prawn, Marsupenaeus japonicus, is one of the most cultivated and consumed species of shrimp. However, very few molecular genetic/genomic resources are publically available for it. Thus, the characterization and distribution of simple sequence repeats(SSRs) remains ambiguous and the use of SSR markers in genomic studies and marker-assisted selection is limited. The goal of this study is to characterize and develop genome-wide SSR markers in M. japonicus by genome survey sequencing for application in comparative genomics and breeding. A total of 326 945 perfect SSRs were identified, among which dinucleotide repeats were the most frequent class(44.08%), followed by mononucleotides(29.67%), trinucleotides(18.96%), tetranucleotides(5.66%), hexanucleotides(1.07%), and pentanucleotides(0.56%). In total, 151 541 SSR loci primers were successfully designed. A subset of 30 SSR primer pairs were synthesized and tested in 42 individuals from a wild population, of which 27 loci(90.0%) were successfully amplified with specific products and 24(80.0%) were polymorphic. For the amplified polymorphic loci, the alleles ranged from 5 to 17(with an average of 9.63), and the average PIC value was 0.796. A total of 58 256 SSR-containing sequences had significant Gene Ontology annotation; these are good functional molecular marker candidates for association studies and comparative genomic analysis. The newly identified SSRs significantly contribute to the M. japonicus genomic resources and will facilitate a number of genetic and genomic studies, including high density linkage mapping, genome-wide association analysis, marker-aided selection, comparative genomics analysis, population genetics, and evolution.

Transcriptome of hepatopancreas in kuruma shrimp Marsupenaeus japonicus under low-salinity stress

The kuruma shrimp Marsupenaeus japonicus is one of the most commercially important shrimp species in the world.Low salinity would aff ect the penetration and immunity,and even led to its death of the shrimp.However,little is known about the molecular mechanism of the effect.Therefore,hepatopancreas of M.japonicus reared under low-salinity stress for 6,12,24,48,and 96 h was analyzed and the results were compared with that of the control group using transcriptomics.After removing reads containing adapters,88890960-1051300444 clean reads were generated from 10 libraries in the control group and experimental group.Compared with the control group,811,589,1095,745,and 875 diff erentially expressed genes were obtained in the five treatment groups.The N50 and N90 lengths of the transcripts were 1746 bp and 436 bp,respectively.The top 20 gene ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways associated with the diff erentially expressed genes were related mainly to osmotic regulation(ion exchange,lipid metabolism and carbohydrate metabolism),immune regulation(cellular and humoral immunity),chitin metabolism,and related functions.The diff erential expression patterns of nine randomly selected genes were confi rmed by quantitative real-time PCR.This is the fi rst report of osmotic regulation-related genes that are diff erentially expressed under low-salinity stress in the hepatopancreas of M.japonicus.Furthermore,we found that M.japonicus initiated its own immune regulation under low-salinity stress.These results will help elucidating the mechanism of osmotic regulation and immune responses in this shrimp species.

Integrative analysis of transcriptomic profile reveals potential roles of miRNAs in regulating development of Marsupenaeus japonicas

Regulation of microRNAs(miRNAs)on various biological processes has been a surprising and exciting field.Identification of miRNAs is the first step to comprehensively understand their functions.However,attempts on global identification and functional verification of miRNAs are very limited in penaeid shrimp Marsupenaeus japonicus,an economically important aquatic species.By performing an integrated analysis of transcriptomic profile from gastrula embryos of M.japonicus,21conserved miRNAs in M.japonicas(mja-miRNAs),belonging to 19 miRNA families,were identified and characterized.Of the 21 mja-miRNAs,15 miRNAs were successfully verified to be predominantly expressed in gastrula stage,where they displayed dynamic expression patterns compared with those in naupliuin stage.Based on perfect or near-perfect match to target region,120 target genes were predicted at transcriptome-wide level.Noteworthy,gene ontology(GO)classification and metabolic pathway annotation revealed eight targets that were actively involved in developmental processes.Of the predicted miRNA-mRNA pairs,six targets were then randomly selected and experimentally validated by dual luciferase reporter assay,where three pairs were proved with potential targeting activity.Overall,to search for conserved miRNAs potentially involved in early development of M.japonicus,we combined in silico and experimental methods,which can be applied in other organisms as well.Our data implied important roles of miRNAs in the early embryonic development and also suggested the presence of complex miRNA-mRNA functional networks in M.japonicus.

以钩虾为前期饵料的日本囊对虾室内养殖技术

主要介绍了利用钩虾作为前期饵料的日本囊对虾室内养殖技术,包括环境要求、放养密度、投喂方法等。分析了利用钩虾替代(冰)鲜饵料的意义。对室内培育钩虾的方法进行了介绍。

影响日本囊对虾养殖的环境因素分析及其健康养殖技术

水环境一直以来都是影响对虾养殖效益的重要因素之一。该文综述了水体环境中影响日本囊对虾养殖的主要影响因子对其存活、生长、免疫等的影响,并总结了日本囊对虾的关键健康养殖技术,以期为提高日本囊对虾的养殖成功率提供指导和帮助。