水产动物6种主要病原菌与抗血清的免疫交叉反应

采用ELISA、试管凝集、Western-blot等方法,分析了鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(V. harveyi)、溶藻胶弧菌(V. alginolyticus)、副溶血弧菌(V. paraheamolyticus)、迟钝爱德华氏菌(Edwardsiella tarda)和荧光假单胞菌(Psedomonas fluorescens)等水产养殖中主要病原细菌与抗血清之间的免疫交叉反应.结果表明,弧菌属细菌之间的交叉反应程度比较大,而与其他两属的细菌之间存在的交叉反应程度小,或不存在交叉反应;Western-blot分析结果显示,哈维氏弧菌、溶藻胶弧菌和副溶血弧菌抗血清分别与其他3种弧菌在分子量为135.6 kD和121.5 kD;95.6 kD,48.4 kD,39.2 kD和34.9 kD;55.1 kD的蛋白带处存在交叉反应,而这些分子量的蛋白带与其他两属的抗血清均不发生反应.     

基于核酸适配体的PCR法检测溶藻弧菌及其灭活菌

溶藻弧菌(Vibrio alginolyticus)分布广,数量多,发病率高,是水产养殖中常见的条件致病菌,而对溶藻弧菌进行快速准确的识别鉴定是其病害防治的前提和基础。核酸适配体,因为具有较高的亲和特异性,在微生物的识别鉴定方面展现出了巨大的优势。本文利用核酸适配体和适配体筛选产物,通过结合、洗涤、加热分离、PCR扩增以及电泳检测等步骤,对溶藻弧菌进行了检测鉴定。结果表明,适配体和筛选产物都能对溶藻弧菌及其灭活菌进行较好的识别鉴定,适配体筛选产物对溶藻弧菌的检测下限为10~3cfu/mL,而对其灭活菌的检测下限为10~2cfu/mL,适配体对溶藻弧菌及其灭活菌的检测下限都可达到10 cfu/mL。该方法对溶藻弧菌有较好的亲和特异性,并能较好地区分溶藻弧菌与哈维氏弧菌等水产常见病原菌,在水产病害的检测中显示了较好的应用前景。     

Odor Modification in Salmon Hydrolysates Using the Maillard Reaction

The aim of this work was to study the effect of adding sugar during proteolysis to promote the Maillard reaction and mask the initial fish odor and off-flavors generated. An experimental design, based on the Doehlert plan, was used to study the influence of hydrolysis conditions (time, temperature, sugar, and antioxidant addition) on the odor characteristics of hydrolysates, soluble protein levels, and amino acid content. Results showed that the lowest level of sugar (10 g of D-xylose added to 1 kg of by-products) was enough to develop a grilled odor in hydrolysates. In the hydrolysis conditions used—i.e., enzyme inactivation at 95°C for 30 min—hydrolysis temperature had no effect on grilled odor production but significantly affected the soluble protein fraction, as did hydrolysis time. Soluble protein content and essential amino acid content increased with the enzymatic reaction but were not modified by adding sugar. Hydrolysis conditions that promote Maillard reactions while keeping a nutritional balance have been identified.     

Degradation and effect of hydrogen peroxide in small‐scale recirculation aquaculture system biofilters

From an environmental point of view, hydrogen peroxide (HP) has beneficial attributes compared with other disinfectants in terms of its ready degradation and neutral by‐products. The rapid degradation of HP can, however, cause difficulties with regard to safe and efficient water treatment when applied in different systems. In this study, we investigated the degradation kinetics of HP in biofilters from water recirculating aquaculture systems (RAS). The potential effect of HP on the nitrification process in the biofilters was also examined. Biofilter elements from two different pilot‐scale RAS were exposed to various HP treatments in batch experiments, and the HP concentration was found to follow an exponential decay. The biofilter ammonia and nitrite oxidation processes showed quick recuperation after exposure to a single dose of HP up to 30 mg L^?1. An average HP concentration of 10–13 mg L^?1 maintained over 3 h had a moderate inhibitory effect on the biofilter elements from one of the RAS with relatively high organic loading, while the nitrification was severely inhibited in the pilot‐scale biofilters from the other RAS with a relatively low organic loading. A pilot‐scale RAS, equipped with two biofilter units, both a moving‐bed (Biomedia) and a fixed‐bed (BIO‐BLOK^®) biofilter, was subjected to an average HP concentration of ～12 mg L^?1 for 3 h. The ammonium‐ and nitrite‐degrading efficiencies of both the Biomedia and the BIO‐BLOK^® filters were drastically reduced. The filters had not reverted to pre‐HP exposure efficiency after 24 h, suggesting a possible long‐term impact on the biofilters.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

中国粳稻春江06抗白背飞虱的系谱分析

对中国粳稻春江06的抗白背飞虱特性进行系谱分析表明,春江06对白背飞虱的拒取食和杀卵抗性均来源于秀水620.秀水620的亲本中,只有秀水04具有较强的拒取食抗性,但没有杀卵抗性.在祥湖24中检测到明显的杀卵反应.亲本秀水04、单209和辐农709具有拒取食抗性,但测21没有.单209和辐农709的共同亲本农虎6号也具有拒取食抗性.然而,农虎6号、农垦58(日本粳稻)和老虎稻(中国粳稻地方品种)不具有拒取食抗性,在田间表现出感虫性.农虎6号、单209、辐农709和秀水04表现出稳定的田间抗性.春江06育种中的两个籼稻品种IR26和IR28高感白背飞虱,既无拒取食抗性,也无杀卵作用.     

农杆菌介导的转高赖氨酸蛋白基因（sb401）水稻T4代分析

通过农杆菌介导的方法用富赖氨酸蛋白基因（sb401）转化粳稻品种日本晴,获得了独立的10个转化株系,对转化株系进行连续自交,通过筛选得到9个纯合的T4代转化株系。 Southern blotting分析发现,整合位点是随机的,并为低拷贝（1~3个）。TAI-PCR扩增得到8个T-DNA侧翼序列,并定位于日本晴的7条染色体上。蛋白质和氨基酸测定分析发现,sb401基因对各株系的蛋白质、赖氨酸和其他氨基酸组分的提高起到了一定的作用。将杂交结果与T-DNA插入位置结合分析发现,在低拷贝的情况下,表达量的差异不明显。     

苎麻属植物RAPD反应体系影响因子的研究

以苎麻属植物中的4个种为材料，用SDS和CTAB2种方法提取基因组DNA并比较其RAPD分析的效果；对RAPD反应体系中的一些重要参数进行了优化，建立了适合苎麻属植物RAPD分析的反应体系。即在25μL的反应总体积中Mg^2 浓度为1.5mmol/L，dNTPs浓度为0.20mmol/L，40ng模板DNA，Taq酶1.0单位；反应程序为95℃预变性5min；前5个循环为94℃变性1min，36℃结合1min，72℃延伸2min；后40个循环为94℃30s，℃36℃1min，72℃2min（最后1个循环为10min）；共45个循环。该反应体系具有良好的稳定性和重复性。