A Shrimp C-type Lectin Inhibits Proliferation of the Hemolymph Microbiota by Maintaining the Expression of Antimicrobial Peptides

Some aquatic invertebrates such as shrimp contain low albeit stable numbers of bacteria in the circulating hemolymph. The proliferation of this hemolymph microbiota in such a nutrient-rich environment is tightly controlled in healthy animals, but the mechanisms responsible had remained elusive. In the present study, we report a C-type lectin (MjHeCL) from the kuruma shrimp (Marsupenaeus japonicus) that participates in restraining the hemolymph microbiota. Although the expression of MjHeCL did not seem to be modulated by bacterial challenge, the down-regulation of its expression by RNA interference led to proliferation of the hemolymph microbiota, ultimately resulting in shrimp death. This phenotype was rescued by the injection of recombinant MjHeCL, which restored the healthy status of the knockdown shrimp. A mechanistic analysis revealed that MjHeCL inhibited bacterial proliferation by modulating the expression of antimicrobial peptides. The key function of MjHeCL in the shrimp immune homeostasis might be related to its broader recognition spectrum of the hemolymph microbiota components than other lectins. Our study demonstrates the role of MjHeCL in maintaining the healthy status of shrimp and provides new insight into the biological significance of C-type lectins, a diversified and abundant lectin family in invertebrate species.     

大兴安岭冻土湿地植物群落结构的环境梯度分析

采用除趋势典范对应分析法(DCCA),对大兴安岭24个冻土湿地植物群落进行排序,对划分出的3个不同类型多年冻土区的湿地植物群落的物种多样性,以及生活型和水分生态型等生态特征进行了环境梯度分析。结果表明:随着纬度的降低,年平均气温和年平均日照时间逐渐增大,年平均降水量和年平均湿度逐渐减小,调查的24个冻土湿地可划分为3组。从大片多年冻土区到大片-岛状多年冻土区,再到稀疏岛状多年冻土区,灌木多样性逐渐减小(p0.05),草本植物多样性逐渐增加(p0.05);中生植物种类所占比例分别为38.5%、55.9%和64.4%,沼生植物的重要值逐渐减小(p0.05),中生和旱生植物的重要值逐渐增大(p0.05),表明冻土湿地的土壤水分条件逐渐中生化,甚至出现旱生化趋势。地面芽植物在3个多年冻土区的湿地植物群落中始终占优势地位,高位芽和地下芽植物次之,反映了该区冬季漫长、严寒而潮湿,夏季较短的气候特征。大片-岛状多年冻土区作为大片多年冻土区和稀疏岛状多年冻土区的过渡区,其湿地植物群落的物种多样性和生态特征与稀疏岛状多年冻土区更相近。     

猪α干扰素重组腺病毒的构建及其抗口蹄疫病毒活性研究

本研究利用RT-PCR方法扩增猪α干扰素成熟蛋白基因后构建了重组腺病毒质粒pAd-poIFN-α,经PacI酶切后转染HEK-293A细胞,3次噬斑纯化后获得了重组腺病毒rAd-poIFN-α。该重组腺病毒于HEK-293A细胞连续传代至20代滴度稳定,为107TCID50/mL。RT-PCR检测证明目的基因在mRNA水平上可有效表达;在PK-15细胞上可以检测到较强的抗猪口蹄疫病毒活性,从而为研究猪口蹄疫免疫防治新技术奠定了重要基础。     

Pattern recognition receptors acting in innate immune system of shrimp against pathogen infections

Invertebrates, including shrimp, have developed very complicated innate immune system against pathogens. Much work has been performed on the innate immunity of shrimp, including immune recognition, signal transduction, effector molecules and antiviral responses due to its great economic value. Pattern recognition is the first step of innate immunity. Pattern recognition receptors (PRRs) sense the presence of infection and activate immune responses. The studies on shrimp PRRs revealed the recognition mechanism of shrimp at a certain degree. To date, 11 types of pattern recognition receptors (PRRs) have been identified in shrimp, namely, β-1,3-glucanase-related proteins, β-1,3-glucan-binding proteins, C-type lectins, scavenger receptors, galectins, fibrinogen-related proteins, thioester-containing protein, Down syndrome cell adhesion molecule, serine protease homologs, trans-activation response RNA-binding protein and Toll like receptors. A number of PRRs have been functionally studied and have been found to have different binding specificities and immune functions. The present review aims to summarize the current knowledge on the PRRs of shrimp.     

Integration of a microbial fuel cell with activated sludge process for energy-saving wastewater treatment: taking a sequencing batch reactor as an example

In the research and application of microbial fuel cell (MFC), how to incorporate MFCs into current wastewater infrastructure is an importance issue. Here, we report a novel strategy of integrating an MFC into a sequencing batch reactor (SBR) to test the energy production and the chemical oxygen demand (COD) removal. The membrane-less biocathode MFC is integrated with the SBR to recover energy from the aeration in the form of electricity and thus reduce the SBR operation costs. In a lab-scale integrated SBR-MFC system, the maximum power production of the MFC was 2.34 W/m(3) for one typical cycle and the current density reached up to 14 A/m(3) . As a result, the MFC contributed to the 18.7% COD consumption of the integrated system and also recovered energy from the aeration tank with a volume fraction of only 12% of the SBR. Our strategy provides a feasible and effective energy-saving and -recovering solution to upgrade the existing activated sludge processes.     

C-type lectin from red swamp crayfish Procambarus clarkii participates in cellular immune response

Lectins are potential immune recognition proteins. In this study, a novel C-type lectin (Pc-Lec1) is reported in freshwater crayfish Procambarus clarkii. Pc-Lec1 encodes a protein of 163 amino acids with a putative signal peptide and a single carbohydrate recognition domain. It was constitutively expressed in various tissues of a normal crayfish, especially in the hepatopancreas and gills. Expressions of Pc-Lec1 were up-regulated in the hepatopancreas and gills of crayfish challenged with Vibrio anguillarum, Staphylococcus aureus, or the white spot syndrome virus. Recombinant mature Pc-Lec1 bound bacteria and polysaccharides (peptidoglycan, lipoteichoic acid, and lipopolysaccharide) but did not agglutinate bacteria. Pc-Lec1 enhanced hemocyte encapsulation of the sepharose beads in vitro, and the blocking of beads by a polyclonal antibody inhibited encapsulation. Pc-Lec1 promoted clearance of V. anguillarum in vivo. These results suggest that Pc-Lec1 is a pattern recognition receptor and participates in cellular immune response. Pc-Lec1 performs its function as an opsonin by enhancing the encapsulation or clearance of pathogenic bacteria.     

Characterization of a novel alpha1,2-fucosyltransferase of Escherichia coli O128:b12 and functional investigation of its common motif

The wbsJ gene from Escherichia coli O128:B12 encodes an alpha1,2-fucosyltransferase responsible for adding a fucose onto the galactose residue of the O-antigen repeating unit via an alpha1,2 linkage. The wbsJ gene was overexpressed in E. coli BL21 (DE3) as a fusion protein with glutathione S-transferase (GST) at its N-terminus. GST-WbsJ fusion protein was purified to homogeneity via GST affinity chromatography followed by size exclusion chromatography. The enzyme showed broad acceptor specificity with Galbeta1,3GalNAc (T antigen), Galbeta1,4Man and Galbeta1,4Glc (lactose) being better acceptors than Galbeta-O-Me and galactose. Galbeta1,4Fru (lactulose), a natural sugar, was furthermore found to be the best acceptor for GST-WbsJ with a reaction rate four times faster than that of lactose. Kinetic studies showed that GST-WbsJ has a higher affinity for lactose than lactulose with apparent Km values of 7.81 mM and 13.26 mM, respectively. However, the kcat/appKm value of lactose (6.36 M(-1) x min(-1)) is two times lower than that of lactulose (13.39 M(-1) x min(-1)). In addition, the alpha1,2-fucosyltransferase activity of GST-WbsJ was found to be independent of divalent metal ions such as Mn2+ or Mg2+. This activity was competitively inhibited by GDP with a Ki value of 1.41 mM. Site-directed mutagenesis and a GDP-bead binding assay were also performed to investigate the functions of the highly conserved motif H152xR154R155xD157. In contrast to alpha1,6-fucosyltransferases, none of the mutants of WbsJ within this motif exhibited a complete loss of enzyme activity. However, residues R154 and D157 were found to play critical roles in donor binding and enzyme activity. The results suggest that the common motif shared by both alpha1,2-fucosyltransferases and alpha1,6-fucosyltransferases have similar functions. Enzymatic synthesis of fucosylated sugars in milligram scale was successfully performed using Galbeta-O-Me and Galbeta1,4Glcbeta-N3 as acceptors.