中国明对虾血蓝蛋白C末端片段在毕赤酵母中的表达及其抗菌活性

为研究中国明对虾(Fenneropenaeus chinensis)血蓝蛋白C末端(FcHC-C)的抗菌功能,将血蓝蛋白基因FcHC的2个C末端基因片段连接到毕赤酵母表达载体pPIC9K中,构建酵母表达载体pPIC9K/FcHC-C。该载体经SalI酶切后,采用PEG法转化毕赤酵母(Pichia pastoris GS115)。转化子经过PCR鉴定后,阳性克隆通过含有G418的YPD平板筛选,获得高拷贝重组子。重组毕赤酵母利用甲醇诱导表达目的基因。经Tricine-SDS-PAGE和Western blot分析结果表明,利用酵母工程菌成功表达了血蓝蛋白C末端片段(rFcHC-C1和rFcHC-C2)。抑菌活性鉴定实验结果显示,重组蛋白rFcHC-C1和rFcHC-C2作为阴离子抗菌肽具有抗真菌和抗细菌的活性。     

Peptidomic analysis of Chinese shrimp (Fenneropenaeus chinensis) hemolymph by magnetic bead-based MALDI-TOF MS

Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based purification(ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) to characterize shrimp hemolymph peptides.Shrimp serum and plasma were used as the source of samples for comparative analysis,and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis.To screen potential specific biomarkers in serum of immune-challenged shrimps,we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps.The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software.Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide(LPS)-infection.The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%.Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph,and will help to enable a better understanding of the innate immune response of shrimps.     

基于太平洋海温的冬小麦春季湿渍害预测模型

为了进一步开展冬小麦春季湿渍害的预测预报业务工作,利用江苏省1969-2008 年60 个气象站常规观测资料和冬小麦产量资料,根据场相关分析原理,将江苏省春季湿渍害3 个风险区的湿渍害指数与太平洋海温资料进行相关普查,结果表明:3 个风险区的小麦春季湿渍害指数和西太平洋海温均存在高相关区,主要位于Nino 区和西太平洋北部海区,其相关程度随时间具有一定的变化趋势;挑选出与湿渍害指数相关的强信号海区作为预报因子,通过最优化相关处理提高预报因子相关性;经稳定性和独立性检验,创建了对应3 个风险区的冬小麦湿渍害指数预测模型,所有模型均通过0.01 显著性水平检验,拟合及试报结果理想,表明预测模型具有一定应用价值。     

Analysis of the Trends of Thunderstorms in 1951–2007 in Jiangsu Province

Based on the 1951–2007 thunderstorms in Jiangsu, a study is conducted for their climate trends, periodicity, spatiotemporal patterns, and the distributions of the first and last days of the thunderstorms at different guarantee rates (GRs) using climate tendency rate, wavelet analysis, and GR for diagnosis. Results suggest that the inter-annual number of thunderstorm days (TSDs) exhibits a decreasing trend in this province. The trend is displayed mainly in the decreasing TSD number in summer and autumn except in spring, when the variation is not significant in the study period. In this province, the TSD number declines by ~2 days per 10 years. On an inter-annual basis, the pronounced positive departures of the number take place chiefly in the early 1960s, the late 1960s to the early-mid-1970s, the late 1980s, and the late 1990s compared with the negative anomalies dominant in the late 1970s to the mid-1980s, the mid-to-late-1990s, and the late 1990s to 2007. There are vast differences in the initial and ending days at diverse GRs in different areas of the province. At 50% GR, the earliest (last) days occur from mid-March to early April (early to late September) while at 80% GR, the initial (last) days are from late March to early May (early to late October). For the distribution of periods, the periods >8–10 years are relatively stable for the entire province. Based on 1951–2007 period analysis, the region north (south) of the Huaihe River experiences TSDs less (more) than normal days in recent years.     

On the relationship between 100-hPa South Asia High and Mei-yu in Jiangsu province

By analyzing the change of an index for the characteristics of South Asia High and variations of upper-air troughs in 2002–2005,we studied the impact of South Asia high on the beginning and ending of the Mei-yu(i.e.sustained rain corresponding to the ripening season of plum)in Jiangsu province.Statistic verification is conducted on the relationships between the index and the Mei-yu season in 1991–2005 to examine the impacts of the SAH characteristics index on a rain intensity index of Mei-yu and regional distribution of a characteristics index for different annual patterns of Mei-yu.Historical composite is performed of the 100-hPa circulation field for these patterns using the 100-hPa geopotential height of Northern Hemisphere from 2002 to 2005 and 45-year NCEP reanalysis to study the difference in the circulation for different patterns of Mei-yu.Diagnostic and statistic conclusions,which share much in common,have been obtained as follows.(1)The characteristics preceding to and the advancement/retreat of SAH and the movement of westerly troughs are the factors that influence the onset time of the Mei-yu season;after the Mei-yu onset,the progression/withdrawal of SAH and how farther east it extends are determining how long the Mei-yu lasts and when it ends.(2)During the Mei-yu,the general 100-hPa circulation situation and average characteristics of the SAH are well corresponding to the characteristics of the season and annual patterns of Mei-yu.In addition,the averages of the SAH ridgeline and east-extending index for June,July and the Mei-yu season have some implications to the forecast of the index of Mei-yu intensity.These conclusions can be served as powerful means in determining the starting/ending dates, duration and annual pattern of the Mei-yu season.     

过硫酸氢钾复合物对对虾养殖底泥硝化作用、氨氧化微生物丰度和群落结构的影响

采用在模拟池塘中投放过硫酸氢钾复合物（KMPS）进行对比实验的方法,探究KMPS对养殖底质硝化作用的影响。通过对氨氮和亚硝态氮含量的检测,探究对不同时期氮素转化的影响,低频率高剂量投放组中的氨氮和亚硝态氮含量显著降低,而高频率低剂量组中氨氮和亚硝态氮的含量显著上升。高频率低剂量KMPS的投放使氨氧化古菌（AOA）和氨氧化细菌（AOB）丰度降低,而低频率高剂量KMPS的投放不会造成丰度降低,而且还表现出部分时期AOB丰度的上升。进一步对群落结构进行分析发现KMPS的投放使AOA群落中Nitrosopumilu属相对丰度降低,Nitrososphaera属的相对丰度上升,这种相对丰度的变化与KMPS的投放方式无关;但AOB群落受到KMPS投放方式的影响,低频率高剂量的KMPS投放下AOB群落优势属相对丰度显著提高。以上结果均说明低频率高剂量KMPS的投放起到了促进底质硝化作用的效果。同时,可为KMPS用于对虾养殖池塘底质改良开辟一个新的途径。     

对虾病害防控的理论与实践

近年来我国对虾养殖持续保持较高增长，但是高密度集约化养殖带来的病害频发和防控手段缺乏等问题，往往给对虾养殖业造成巨大损失，使对虾养殖业成为一个高风险高回报的产业。针对我国对虾养殖业存在的主要危害因子如病原微生物、真菌毒素和环境胁迫等，笔者所在研究团队多年来从对虾免疫反应和生长代谢调控角度，探讨了对虾应对外界刺激的内在机制。同时，研究开发了水产专用的微生物制剂、抗病功能蛋白以及生物饲料等新型的安全投入品，开展了从基础理论到实际应用的系统性研究工作。本文综述了研究团队多年来在对虾病害防控的理论与实践方面的研究成果。     

饲料中添加两株乳酸菌及其发酵上清液对凡纳滨对虾消化酶活性的影响

为了探讨前期实验获得的益生菌及其发酵上清液对凡纳滨对虾消化功能的影响,将初始体质量3.5 g±0.06 g的凡纳滨对虾(Litopenaeus vannamei)在30℃±2℃环境下于水族箱中养殖4周。以基础饲料为对照组,在基础饲料中分别添加戊糖乳杆菌(Lactobacillus pentosus)HC-2、粪肠球菌(Enterococcus faecium)NRW-2、戊糖乳杆菌HC-2的发酵上清液,最终每克基础饲料中含有1.0×107 CFU益生菌或此菌量所对应的发酵上清液,共配制3组实验饲料。实验结果如下:与对照组相比,各实验组对虾肠道中蛋白酶活力显著提高(P0.05),但肝胰腺中蛋白酶活力无显著差异;添加NRW-2的实验组与对照组相比,肠道和肝胰腺中淀粉酶和脂肪酶活力均显著提高(P0.05),HC-2组对虾肝胰腺中淀粉酶及肠道中脂肪酶的活力显著提高(P0.05),而HC-2发酵上清液的添加仅显著提高了对虾肠道蛋白酶和淀粉酶的活性。结果表明饲料中添加NRW-2、HC-2及其发酵上清液可以提高凡纳滨对虾肠道及肝胰腺中消化酶的活力,但在不同组织中提高消化酶活性的种类是不同的,为水产养殖安全投入品的开发及乳酸菌在水产养殖中的推广应用提供一定的科学依据。     

Low temperature stress on the hematological parameters and <Emphasis Type="Italic">HSP</Emphasis> gene expression in the turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>

To study the effect of low temperature stress on hematological parameters and HSP gene expression in the turbot (Scophthalmus maximus), water temperature was lowered rapidly from 18 to 1°C. During the cooling process, three individuals were removed from culture tanks at 18, 13, 8, 5, 3, and 1°C. Blood samples and tissues were taken from each individual, hematological indices and HSP gene expression in tissues were measured. The red blood cell count, white blood cell count, and hemoglobin concentration decreased significantly (P < 0.05) as temperature decreased. Enzyme activities of plasma alanine transaminase and creatine kinase increased as temperature decreased, whereas aspartic transaminase and γ-glutamyl transpeptidase activities displayed no obvious changes above 1°C and lactate dehydrogenase activity increased first and then decreased. Blood urea nitrogen and uric acid levels were highest at 8°C, and creatinine concentration was highest at 3°C. The concentrations of plasma cortisol, cholesterol, and triglyceride all increased significantly (P < 0.05) as temperature decreased. The serum glucose concentration increased first and then decreased to the initial level. The HSP70 mRNA expression showed various patterns in different tissues, whereas HSP90 mRNA expression showed the same tendency in all tissues. Overall, these results indicate that temperature decreases in the range of 8 to 5°C may induce a stress response in S. maximus and that temperature should be kept above 8°C in the aquaculture setting to avoid damage to the fish.