Comparative study on the allergenicity of different Litopenaeus vannamei extract solutions

Allergen extracts are widely used for allergy diagnosis and treatment. The application of shrimp extract is hampered due to the low protein concentration and the inconsistent allergenicity. Extracting solutions are considered to be the primary limiting factor of protein extraction from crustaceans. This study aimed to select an optimal solution for shrimp protein extraction by comparing the allergenicity of different shrimp extracts. The effect of 7 existing or modified extracting solutions were evaluated, including the glycerol-NaCl solution, the glycerol Cocaine's solution, the buffered saline solution, the Cocaine's solution, the Glucose leaching solution, 1 mol L-1 KCl solution, and 0.01 mol L-1 phosphate buffered saline solution with and without dithiothreitolor(DTT). The quantitative(protein concentration) and qualitative parameters(SDS-PAGE protein patterns and immuno-reactivity) were determined using the sodium dodecyl sulfate polyacrylamide gel electrophoresis, enzyme linked immunosorbent assay and immunoblotting assay. Results showed that the 1 mol L-1 KCl solution with DTT was optimal for shrimp protein extraction, which yielded high concentration and allergenicity in the protein extract, including major and minor allergens. The 1 mol L-1 KCl solution with DDT is proposed for preparation of shrimp extract and associated allergy diagnosis, as well as potential applications for other crustaceans.     

凡纳对虾人工繁殖的初步研究

研究了海水比重的变化对凡纳对虾亲虾人工繁殖活动的影响及换水对亲虾交配率的影响。试验采用剪眼柄、强化培育、控光、换水等技术措施 ,将凡纳对虾培育成熟并使之自然交配产卵 ,孵化出无节幼体 ;亲虾的平均产卵量可达 1 6万粒左右 ,卵的受精孵化率可达 80 %以上。结果表明 :适时换水可将亲虾的交配率从 4 6 %提高到 79% ;凡纳对虾人工繁殖的适宜海水相对密度为 1 0 1 6以上。     

凡纳滨对虾性别分化的初步研究

通过组织学连续切片与扫描电镜观察,研究凡纳滨对虾的性别分化。结果显示:凡纳滨对虾外部形态的性别分化最早发生在仔虾后第37天,此时在扫描电镜下可观察到雄性附肢的芽基,仔虾后第45天,光镜下可观察到交接器;仔虾后第24天出现原始生殖细胞(PGCS),位于背大动脉的两侧及围心腔膜的腹面,细胞个体及细胞核明显比周围的细胞大,具有强嗜碱性,细胞核圆形或椭圆形,核内染色质清晰可见,随着生殖腺的发育,在第43天出现生殖管,第55天生殖腺可区分为精巢和卵巢。     

A Comparative study on the nonspecific immunity of juvenile Litopenaeus vannamei ever inhabiting freshwater and seawater

A study on the nonspecific immunity of Litopenaeus vannamei ever inhabiting freshwater and seawater was carried out at different molt stages by comparing their total hemocyte count(THC) and respiratory burst(RB) and activity of phenol oxidase(PO), nitric oxide synthase(NOS) and lysozyme(LY). Two-way ANOVA showed that salinity and molt stage independently affected THC and RB and the activity of PO, NOS and LY of juvenile L. vannamei significantly(P 0.05). The THC and RB and the activity of NOS gradually increased from the post-molt stages(A and B) to the pre-molt stages(D0–D3), which were common in shrimps inhabiting freshwater and seawater. The activity of PO peaked at the inter-molt stage(C), and touched the lowest at the post-molt stage in freshwater and pre-molt stage in seawater. The activity of LY was stable over the molt cycle. The RB and the activity of PO, NOS and LY of juvenile L. vannamei were significantly lower in freshwater than in seawater; whereas THC was significantly higher in freshwater than in seawater(P 0.05). It was concluded that the post-molt stage(especially stage A) was critical to shrimp culture, which should be intensively attended when L. vannamei was cultured in freshwater.     

凡纳滨对虾快速生长家系选育的初步研究

采用群体选择和家系选择相结合的方法,收集有代表性的优良群体103个,经过2代的群体选育,保留5个基础群体进行家系选育,育出全同胞家系180个,半同胞家系60个,选留其中的35个家系作为下一代的亲本,选育的D2家系生长速度与进口F1代相接近,养殖有效积温2 000℃时体重分别为26.17 g和26.47 g,抗WSSV能力较进口F1代强,感染WSSV 5 d死亡率分别为53.57%和72.73%。提出用等同的有效积温的体重来比较不同家系的生长,用生长速度、存活率和个体均匀度作为家系选育的指标。     

Supplementation of Yeast Extract to Practical Diet Improves the Growth,Anti-Oxidative Capacity and Intestinal Morphology of Shrimp Litopenaeus vannamei

Two control diets based on the commercial formula were designed to contain high(27%, D1) and low(22%, D2) levels of fish meal, respectively. Into D2, 500, 1000 and 1500 mg kg~(-1) of yeast extract were added, respectively, yielding three experimental diets(YE1 through YE3). Shrimp(initial body weight 0.30 g ± 0.02 g) were fed with the experimental diets, five tanks each diet and 30 shrimp individuals each tank, for 8 weeks, and then challenged with Vibrio parahaemolyticus. The results showed that the specific growth rate(SGR) of shrimp in D2 was significantly lower than that of shrimp in D1(P 0.05). The SGR of shrimp in YE3 was similar to that of shrimp in D1. The feed intake of shrimp was similar between D1 and D2. The feed conversion ratio and protein efficiency ratio of shrimp were similar among all diets(P 0.05). YE significantly improved the activity of glutathione S-transferase. The concentration of glutathione(GSH) and the total serum anti-oxidative capacity(T-AOC) of D1 were significantly higher than those of shrimp feeding other diets(P 0.05). The content of serum malondialdehyde of shrimp feeding YE2 and YE3 was significantly lower than that of shrimp feeding D2(P 0.05). The thickness of intestine muscular layer of shrimp feeding YE1 and YE2 was similar to that of shrimp feeding D1. The shrimp feeding YE1 showed the highest villus height of intestine among all groups. The cumulative mortality after challenging was similar among all groups(70.00%–86.67%)(P 0.05). In conclusion, 1000–1500 mg kg~(-1) of YE was suggested to be supplemented into the practical diets to improve the growth, anti-oxidative capacity and intestinal morphology of shrimp L. vannamei.     

眼柄粗提物对凡纳滨对虾性分化的影响

采用生化方法制备凡纳滨对虾[Litopenaeus(Penaeus)vannamei)]眼柄粗提物,用活体注射法研究不同粗提物剂量对去眼柄凡纳滨对虾卵母细胞直径大小的影响,以检验其性腺抑制素(Gonadinhibiting hormone,GIH)的生物活性;在凡纳滨对虾仔虾后17、22和27 d,分别以不同浓度眼柄粗提物浸泡24 h,结果显示:除0.001个眼柄/mL的GIH浓度外,0.01、0.1和1个眼柄/mL组均显著降低凡纳滨对虾卵母细胞直径(P﹤0.05);浸泡处理后,仔虾后17 d GIH处理组凡纳滨对虾雌性率达70.1%～80.1%;仔虾后22 d GIH处理组雌性率达65.8%～71.7%,各浓度GIH处理组雌性率比对照组(雌雄比例约为1:1)显著提高(P﹤0.05);仔虾后27 d,0.01和1个眼柄/mL处理组雌性率(约58%)略高于对照组,但0.1个眼柄/mL处理组雌性率(73.1%)与对照组相比显著提高(P﹤0.05)。结果表明较低浓度的眼柄粗提物可明显诱导凡纳滨对虾雌性化,在仔虾后22 d前处理效果最佳。     

淮山提取液替代羧甲基纤维素钠作为酸奶稳定剂的效果研究

采用正交试验等方法对淮山提取液（DB-Ext）代替羧甲基纤维素钠(CMC)作为调配型酸奶稳定剂的效果进行了初步探讨。结果表明以原料的6.0倍水量在110℃的温度条件下，提取40min，所得的淮山提取液的粘度为3500mPa     

凡纳滨对虾不同生长阶段食物组成结构的研究

利用稳定同位素碳比率δ13C作为天然示踪物,对高位养殖池生态系统中人工饲料和天然饵料提供的能源物质对养殖凡纳滨对虾生长的贡献比率进行研究。结果表明:虾体的δ13C值受食物δ13C值的影响,由于其摄食饵料的δ13C值逐渐减小,虾体的δ13C值也逐渐降低,逐渐接近于最后人工饲料的δ13C值,试验虾体的δ13C值由养殖开始时的-19.52‰±0.09‰变为实验结束时的-21.74‰±0.17‰;对虾的食性在10d左右时发生转变,34d左右时人工饲料和浮游动物对虾体的能量贡献基本相同,养殖结束时对虾生长能量的97.29%±0.98%来源于人工饲料。     

《儒林外史》人物对比讽刺谈

《儒林外史》是一部讽刺小说,主要运用对比讽刺,包括“鹤立鸡群”式、“言行相悖”式、“无独有偶”式、“前后矛盾”式、“自打嘴巴”式以及明比、暗比等讽刺。在作者的笔下,讽刺手法剥光了那些令人憎恶的封建文人、官僚豪绅、市井无赖等的外衣,露出了他们丑恶的灵魂,深刻地揭露了行将崩溃的封建制度的腐朽性,强烈地抨击了罪恶的科举制度。这种讽刺艺术在我国古代小说发展史上达到了顶峰。     

空间分析中几种插值方法的比较研究

针对常用的空间插值方法在精度分析方面比较的不足,从地质统计学、数学函数和几何方法方面分别选取普通克里金插值法、三次样条函数法和距离权重倒数法进行插值精度的比较与分析。文章分析了各个方法的优缺点和局限性,选取疏密差异较大的均匀和不均匀离散点数据进行实验,利用三次趋势面模型和MATLAB,ArcGIS,GS+Version...     

酵母培养物对凡纳滨对虾肠道菌群结构的影响

在基础饲料中分别添加质量分数0(对照)和0.30%的酵母培养物,配制2种等氮等脂饲料(分别记为Y0、Y0.3组),投喂初始体质量为(1.20±0.01)g的凡纳滨对虾(Litopenaeus vannamei)56 d。提取凡纳滨对虾肠道总DNA,采用Illumina Mi Seq高通量测序比较样品菌群组成和多样性,研究饲料中添加酵母培养物对凡纳滨对虾肠道菌群结构的影响。结果表明,Y0.3组对虾肠道分类单元(OTUs)数量、丰富度指数(Chao1、ACE值)、多样性指数(Shannon、np Shannon指数)与Y0组差异无统计学意义(P0.05);从物种分类树发现,两组的主要细菌类群均为变形菌门(Proteobacteria),其中,γ-变形菌纲是对虾肠道的最主要菌群,所占比例达69.48%;从科分类水平看,Y0.3组对虾肠道菌群链球菌科(Streptococcaceae)比Y0组提高4.98%;从优势属水平来看,Y0.3组对虾肠道的乳球菌属(Lactococcus)高于Y0组,差异有统计学意义(P0.01);Y0.3组对虾肠道的发光杆菌属(Photobacterium)低于Y0组,差异有统计学意义(P0.05)。饲料中添加质量分数0.30%酵母培养物可改善凡纳滨对虾肠道菌群多样性。     

水平面上太阳辐射空间模拟对比研究

面域的太阳辐射数据是进行其他各项有关辐射研究的基础。由辐射台站的点数据来模拟太阳辐射的空间分布时,传统的模拟方式都会产生精度与效率之间的矛盾。直接以辐射观测值与其影响因子建立非线性回归模型,以各因子的面域数据来模拟水平面上太阳辐射的空间分布,与引入了辐射基数值的模式相比,模型的误差更小,复相关系数大于0．96,运算效率提高29-286倍。当采用模拟的天文辐射经验公式来计算辐射基数时效率仍可提高2-11倍。用两种方式的回归模型模拟中国1km栅格的水平面太阳辐射,平均差值仅2.41％,而本文模式模拟结果与实测值更为接近。     

Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei

The effects of different hydrolysis methods on peptidoglycan(PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp,Litopenaeus vannamei.PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations.A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp.The composition,concentration,and molecular weight ranges of the soluble PG were analyzed.Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment.The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus(WSSV).Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h,and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme.SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods.Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes.In addition,the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV,whereas no increased resistance was observed for preparations containing less low-molecular-weight PG.These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.     

IGS精密星历和钟差的算法比较研究

通过编程对IGS精密星历和钟差的Lagrange插值、Neville插值、线性插值、多项式拟合算法等进行分析。结果表明:对于精密星历,插值算法的精度较相同阶次的拟合算法差,在拟合与插值时,选择最佳的阶数,其精度可达到毫米级;对于钟差,采用线性插值算法更为简单有效。     

凡纳滨对虾促雄性腺的组织学初步观察

通过显微观察和组织学连续切片,对凡纳滨对虾的促雄性腺进行研究。结果表明:凡纳滨对虾的促雄性腺埋于精荚囊端壶腹外侧的肌肉中,呈不规则的块状,Harris苏木精染色呈嗜碱性。H?E染色显示,促雄性腺主要由两种形态的细胞构成,一种核质致密嗜碱性非常强,细胞核被染成深蓝色,核仁很难辨认,大多呈圆形或卵圆形(A型细胞);另一种细胞多为卵圆形,细胞核较大,核质疏松,被染成浅蓝色,核仁1～2个清晰可见(B型细胞)。此外,促雄性腺中A、B细胞的出现及其数量随精巢发育而发生变化,繁殖完成后细胞解体。     

凡纳滨对虾潘状幼体厌食症的防治

潘状幼体厌食症是指潘状幼体发育到第2期后,出现摄食突然减少和停食的现象,幼体发病率和死亡率较高。探讨育苗用水处理方法、抗菌素、有益微生物制剂对防治凡纳滨对虾潘状幼体厌食症的效果。结果表明：1）用次氯酸钠、百碘、溴氯海困对育苗用水进行消毒处理没有预防潘状幼体厌食症作用,用甲醛处理水有一定的预防效果,幼体发病率为75.3％,低于对照组的88.2％,差异有统计学意义（P〈0.05）;2）利福平、土霉素、庆大霉素、诺氟沙星、复方新诺明等抗菌药物对潘状幼体厌食症防治效果不佳,除利福平外,其他药物组幼体发病率、存活率与对照组差异无统计学意义（P〉0.05）;3）有益微生物制剂光合细菌、乳酸杆菌、芽孢杆菌、复合活菌制剂EM活菌、虾蟹宝活菌和乐多源活菌对潘状幼体厌食症都有不同程度的预防作用,幼体发病率、存活率与对照组差异有统计学意义（P〈0.05）,以乐多源活菌预防效果最好,幼体发病率仅19.8％,比对照组低63.3％,治疗作用仅乐多源活菌较好,可治愈50.0％的发病幼体,其他活菌治疗效果不佳。建议潘状幼体厌食症应采取处理水和使用有益微生物制剂相结合方法进行防治。     

潜水含水层井流模型的对比研究

依托在潜水含水层中开展的10组高分辨率抽水试验,探讨了泰斯(Theis)、博尔顿(Boulton)和纽曼(Neuman)3种潜水井流模型的识别精度。结果表明:泰斯模型由于忽略了潜水含水层的三维流动和潜水含水层重力释水的滞后效应,因此无法真实反映潜水井流的运动特征和释水机理;博尔顿模型能够较好地刻画抽水试验早期潜水含水层重力给水滞后的主要特征;既考虑潜水含水层重力滞后给水机理,又同时考虑了潜水含水层三维流动特征的纽曼模型最能全面反映潜水含水层井流试验的运动特征和释水机理。      

抗菌肽对凡纳滨对虾生长和机体免疫的影响

基础饲料(对照组)中添加10g/kg抗菌肽制剂喂养体重0.10±0.01g、体长2.17±0.11cm的凡纳滨对虾(Penaeus vannamei)30d,进行10d的水浴攻毒(鳗弧菌10~8/mL),探讨饲料中添加抗菌肽对凡纳滨虾生长、成活率、免疫保护率,以及肌肉中溶菌酶活性、超氧化物歧化酶活性、总抗氧化能力的影响。结果表明:饲料中添加抗菌肽可显著提高凡纳滨对虾相对增重率、成活率和对致病菌的免疫保护率(P<0.05),但对饲料系数无显著影响(P>0.05);未受外界微生物刺激时,饲料中添加抗菌肽对凡纳滨对虾总抗氧化能力有促进作用(P<0.05),受外界微生物刺激后,饲料中添加抗菌肽喂养凡纳滨对虾可提高对虾超氧化物歧化酶活性(P<0.05)和总抗氧化能力(P<0.01)。因此饲料中添加抗菌肽喂养凡纳滨对虾可提高其机体非特异性免疫。     

Influence of Ultrasonic Treatment on the Allergenic Properties of Shrimp （Penaeus vannamei ） Allergen

The present study was undertaken to determine whether high intensity ultrasound could reduce the allergic properties of shrimp allergens. Reducing the allergenic properties of these allergens will be beneficial to allergic individuals. Samples of shrimp protein extract and shrimp muscle were treated by high-intensity ultrasound with water bathing at 0 ℃ or 50 ℃ for different time periods. The treated and untreated samples were then analyzed by SDS-PAGE, Western blots and competitive inhibition ELISA （Ci-ELISA） to determine the shrimp allergenicity. The results show that high-intensity ultrasound has no effect on allergenicity when the extracts were treated at 0℃. However, a significant decrease was observed in the level of the major shrimp allergen, Pen a 1, when the samples were treated at 50 ℃. In the determination of allergenicity with CiELISA, a reduction in IgE binding was also observed.