中华绒螯蟹多倍体的诱导研究I.细胞松驰素B诱导中华绒螯蟹三倍体和四倍体胚胎

采用细胞松驰素Ｂ（Ｃ．Ｂ）处理法，研究了诱导中华绒螯蟹（Ｅｒｉｏｃｈｅｉｒｓｉｎｅｎｓｉｓ）三倍体、四倍体的可能性和处理条件。实验所用Ｃ．Ｂ浓度为０．５～２．０ｍｇ／Ｌ，处理的起始时间分别在受精后１０～２５ｍｉｎ（诱导三倍体）和８．５～９．８ｈ（诱导四倍体），处理的持续时间为１０～２５ｍｉｎ。三倍体的适宜诱导条件为卵子在受精后１５ｍｉｎ，浓度为１．５ｍｇ／Ｌ的Ｃ．Ｂ溶液处理１８ｍｉｎ。三倍体诱导率最高可达５８．１８％。根据诱导三倍体的适宜Ｃ．Ｂ处理浓度和处理时间，用１．５ｍｇ／ＬＣ．Ｂ溶液处理卵子１８ｍｉｎ，获得了２９．４６％～５７．８９％的四倍体胚胎。另外，还对中华绒螯蟹三倍体、四倍体胚胎的诱导技术、诱导参数对胚胎发育的影响等有关问题进行了分析讨论     

甲壳类动物颗粒饵料制造的关键

近年来，水产养殖作为世界市场中多种产品的来源已占据重要的地位。但由于饵料费用占产品费用很大的一部分，所以始终都将研究集中在使用简单、有效、廉价的饵料成分和饵料工艺来生产人工饵料上。目前的研究以影响颗粒饵料在水中稳定度和获得可用碾磨颗粒饵料的关键作为设计的标准。采用优质工程设计按５个因子、２个水平来验实，其５个因子、２个水平为：粘合剂成分（面粉／土豆粉）、粉末混合和温混合的时间（１０～１５ｍｉｎ）、     

3种饵料金藻的超低温保存研究↑（*）

用两步冷冻法研究绿色巴夫藻（Ｐａｖｌｏｖａｖｉｒｉｄｉｓ）、湛江等鞭金藻（Ｉｓｏｃｈｒｙｓｉｓｚｈａｎｊｉａｎｇｅｎｓｉｓ）、球等鞭金藻（Ｉ．ｇａｌｂａｎａ３０１１）的超低温保存（液氮,－１９６℃）,探讨了影响存活率的内、外因素。结果表明,降温速率０．７℃／ｍｉｎ,预冻温度－４０℃,预冻时间３０ｍｉｎ时可获得较高的存活率。１５％ＤＭＳＯ或１５％ＤＭＳＯ＋１０％蔗糖是良好的抗冻保护剂。某些藻类在生长静止期或在低于最适温度下培养时可显著提高其存活率。通过控制内、外条件,３种金藻的存活率都可达到３０％以上,比以往的报道有显著提高。     

泥鳅人工繁殖最适条件初探

本文对影响泥鳅人工繁殖主要外界因素进行了研究。结果表明，使用促黄体生成素释放激素类似物对泥鳅进行人工催产，剂量为３０毫克／尾（雄鱼减半），水体ｐＨ值为６．５￣７．０，性比（♀：♂）为１：２，下午６时注射，水温在２０℃￣３０℃时，催产效果最佳。     

海产饵料微藻超低温保种技术的研究↑（*）

１５种海产饵料微藻在超低温（－１９６℃）下保存试验结果表明，除了波海红胞藻和中肋骨条藻未能活化复苏外，其余１３种微藻都能在一定的时间内活化复苏，进行正常的生长繁殖，只是各种微藻的生长延缓期和生长率各有不同。细小裂面藻的延缓期最短，为４天，１５天的平均生长率最大（Ｋ＝０．２５３）；盐沼杜氏藻为最长，１５天，生长率最小（Ｋ＝０．０７７），其他微藻的延缓期都在５—１０天，生长率为０．１２１－０．２０４。多数微藻在超低温条件都能长期保持生命活力，超低温技术是微藻保种中的一种较好的方法。     

黄霉素对生长缓慢的欧洲鳗促生长效果的研究

本文讨论了在配合饲料中添加不同剂量黄霉素对生长缓慢的欧洲鳗（Ａｎｇｕｉｕａａｎｇｕｉｌｌａ）仔鳗的促生长效果。试验结论：添加黄霉素的配合饲料对欧洲鳗有显著促生长作用。其适宜添加量为１０－１５ｐｐｍ，以添加１５ｐｐｍ为最佳，该组的增重比为１２１（对照组为１００），饵料系数为１．４５，经济效益最好。     

黄河滩区泥鳅集约化养殖高产技术

我省范县地处黄河背河洼地，１９９９～２００２年，该县在我站指导下，在陈庄乡、杨集乡等相继建立了６个泥鳅养殖示范基地，进行了大规模的泥鳅养殖。经过几年来的生产实践，亩产均达到１０００千克以上，亩利润达４０００元。现将几年来泥鳅养殖过程中的主要关键技术进行总结。一、泥鳅的苗种培育技术（一）清塘与施基肥１．池塘的清整苗种培育以土池为好，面积以３０～１００米２为宜，池深４０～６０厘米。池中开挖鱼溜，以利其栖息和避暑防寒，池埂池底夯实，进排水口设拦鱼网，池底铺垫１５～２０厘米淤泥层，池中投放浮萍或水草起遮…     

菲律宾蛤仔（Ruditapes philippinarum）低温保活方法的研究↑（*）

本文在菲律宾蛤仔（Ｒｕｄｉｔａｐｅｓ ｐｈｉｌｉｐｐｉｎａｒｕｍ）生物学研究基础上，对菲律宾蛤仔在 ２７． ０～－１． ７℃范围内不同温度条件下的存活率，失重率及保活过程中的化学变化等进行了系统的分析研究，确定了菲律宾蛤仔的保活工艺和方法。结果表明，（１）菲律宾蛤仔的最肥季节为每年的 ３～ ７月，出肉率（干物计）最高可达 １０％；（２）最低保活温度为－１．７℃；（３）－１．０～－１．７℃可保活１３天，存活率９１％；（４）－１．０～１．７℃保活，保活时间最长，失重率也低，主要化学成分无显著变化。     

夏季白令海温跃层和中层冷水的分布特征

研究了夏季白令海阿留中海盆区温跃层和中层冷水的分布特点及形成机制。结果表明,（１）以水温为主要分析指标,调查海区自上而下（０～５００ｍ）层,可分为上匀和层、浊跃层、中居冷水和下匀和层（２）温跃层广泛分布于大部分调查海域,绝大多数测站其深度在２５～５０ｍ,强度在０．１０～０．１６℃／ｍ,其中东南部为一相对强温跃区（≥９０．１５℃／ｍ）;（３）中层冷水的上界深度在４０～１３０ｍ左右,下界深度在１５０～     

罗氏沼虾健康繁殖技术

近年来人工养殖罗氏沼虾出现个体小、性成熟提前、抗病力低等不良趋势，为确保罗氏沼虾后代的优良性状，我们在实践中总结出了罗氏沼虾健康繁殖技术，现将其技术要点总结如下。１亲本养殖池塘条件土池长方形，阳光充足，通风良好，面积０．６～１．３公顷，水深１．５米以上，池底淤泥不超过１５厘米，池底有底栖生物（螺、蚬、蚌等），沉水植物覆盖率１５％～２０％。放养水温稳定在２０℃后放养。当年良种苗投放不超过２．５万尾／６６７米２。放养时先将装苗氧气袋放入池水中缓温１０～２０分钟，再行放养。饵料全程以天然饵料为主（如螺…     

超高压处理对牡蛎(Crassostrea gigas)杀菌及贮藏品质的影响

以牡蛎(Crassostrea gigas)为研究对象,研究了不同超高压处理条件对牡蛎体内微生物的消减作用,分析了冷藏过程中挥发性盐基氮(TVB-N)和 pH 值的变化情况,对比了热加工和超高压处理对牡蛎滋味的影响。结果显示,经300 MPa 处理15 min、400 MPa 处理10、15 min 和500 MPa处理5、10和15 min 后,牡蛎体内的菌落总数均有2个对数以上的减少,表明超高压处理能有效消减牡蛎体内的微生物;牡蛎经300 MPa 处理15 min、400 MPa 处理10 min 和500 MPa 处理5 min,在4℃冷藏条件下保存,货架期延长至15 d;超高压处理能有效抑制牡蛎在4℃贮藏过程中挥发性盐基氮的产生,牡蛎经300 MPa 处理15 min,冷藏20 d 后 TVB-N 值仍小于10 mg/100 g,符合生鲜牡蛎要求;滋味方面,与加热处理相比,超高压处理后牡蛎更接近生鲜牡蛎,尤其300 MPa 处理15 min 的牡蛎同生鲜牡蛎最为接近。研究表明,选取300 MPa 处理15 min 作为牡蛎加工的最佳处理条件,即能有效消减牡蛎体内的微生物,同时能很好地保持牡蛎品质。     

Evaluation of cytochalasin B (CB) treatments for triploidy induction in the blacklip abalone, Haliotis rubra (Leach, 1814)

The effectiveness of cytochalasin B (CB) treatments for inducing triploidy was evaluated in the blacklip abalone Haliotis rubra (Leach, 1814) in two orthogonal design experiments. The first experiment employed three dosages (DSs) of 0.25, 0.5 and 1.0 mg CB L^?1, three starting times (STs) of 5, 15 and25 min post fertilization and three treatment durations (TDs) of 10, 20 and 40 min, for a total of 27 treatments. The second experiment comprised of two DSs of 0.25 and 0.5 mg CB L^?1, five STs of 5, 15, 20, 25and 30 min post fertilization, and three TDs of 10, 20 and 40 min, for a total of 30 treatments. Water temperature was held at 17.5–18.5°C. Day 3 larvae were sampled for triploidy using flow cytometry (FCM) and survival. Optimal inductions were treatments starting at 15 or 20 min post fertilization and continuing for 40 min, and those initiated 25 or 30 min post fertilization for 20 or 40 min, using 0.5 mg CB L^?1. These treatments were all targeted at inhibition of the second polar body (PB2) formation and yielded triploidy rates of 84.8–89.5% coupled with (relative) survival rates of 20.1–52.1% in the first experiment, and corresponding rates of 86.5–96.5% and 33.0–74.1%, respectively, in the second experiment. A common and essential feature of these optimal conditions is that treatment must fully span the period of time for most of the eggs to extrude PB2. Treatments that resulted in suppression of the first polar body (PB1) formation induced triploidy levels below 71.5% and 57.6% in experiments 1 and 2 respectively. Treatments that had overlapping effects on both PB1 and PB2 extrusion led to triploidy rates above 80% but very low survival rates of 1.8% and 5.4% in experiments 1 and 2 respectively.     

热休克诱导马苏大麻哈鱼三倍体的初步研究

为探索马苏大麻哈鱼(Oncorhynchus masou)三倍体育种技术方法,更好地解决其个体小、生长慢和性成熟后死亡率高等问题,采用热休克法进行三倍体诱导实验,设4个诱导温度(24、26、28、30℃),2个起始诱导时间(15 min、20 min)和2个持续诱导时间(15 min、20 min)共分13个实验组和1个对照组。结果显示,13个实验组均能诱导出三倍体个体,而不同温度组的诱导率差异显著,分别为13.3%、31.65%、52.28%和78.81%(P<0.0.5)。随着温度的上升,孵化率呈显著降低的趋势。结果表明,温度是影响诱导成功的关键因素,利用热休克诱导受精卵制备陆封型马苏大麻哈鱼三倍体苗种的方法是可行的,三倍体诱导的最佳条件是:水温28℃,卵子受精后15 min持续处理20 min,发眼率(72.57±0.26)%,孵化率(60.92±0.31)%,三倍体率53.1%,综合诱导效果最佳。     

Induced triploidy in the European clam, Ruditapes decussates (L.), and performance of triploid larvae

Abstract. For the first time, effective treatments using cytochalasin B were developed to induce triploidy in the European clam, Ruditapes decussatus (L.). The percentage of triploid embryos was assessed by karyological or image analysis. Two treatments (0·5 or 1 mg of cytochatasin B (CB) per ml of dimethyisulfoxide in 1 litre of sea water) were applied at different times after fertilization (10 to 25min), for two different periods (15 and 20min). Best results were obtained for a CB concentration of 1 mg/1. When treatment was applied 15min after fertilization for a 20min period, 94% and 95% of triploid embyros were obtained in two repeated experiments. At metamorphosis, the treated larvae appeared to be no smaller than the control larvae in all experiments. However, in general, significantly higher mortalities for CB-treated batches were found when compared with the untreated batch.     

新型笼目海带风味果冻的加工工艺研究

以日本笼目海带为原料进行笼目海带风味果冻加工工艺探究。通过单因素及正交试验确定海带汁的最佳脱腥工艺和复配胶比。研究结果表明,最优脱腥工艺为:绿茶添加量0.15%,脱腥时间25 min,温度90℃;最佳复配胶比例为:卡拉胶∶魔芋胶∶琼脂=10∶2∶1。采用正交试验获得笼目海带风味果冻加工优化工艺为:以20%笼目海带汁为底物,复配胶添加量1.3%(卡拉胶、魔芋胶、琼脂),KCl 0.06%,白砂糖3.0%,浓缩果汁(苹果、橘子)添加量10.0%,pH 4.0。制作的笼目海带果冻色泽均匀,酸甜适中,风味独特,符合国家果冻食品卫生要求(GB 19299-2015)。     

Optimization of triploidy induction in the blacklip abalone, Haliotis rubra (Leach, 1814), using 6-dimethylaminopurine

Optimal conditions of 6‐dimethylaminopurine (6‐DMAP) for triploidy induction in the blacklip abalone Haliotis rubra (Leach, 1814) were investigated, targeting inhibition of second polar body (PB2) formation. Two experiments were conducted at a water temperature of 17.5–18.5°C where factorial combination of (1) four dosages (DSs) of 100, 150, 200 and 250 μM 6‐DMAP, four starting times (STs) of 15, 20, 25 and 30 min post fertilization, and two treatment durations (TDs) of 20 and 30 min and (2) three DSs of 50, 100 and 150 μM 6‐DMAP, three STs of 15, 20 and 25 min post fertilization, and three TDs of 10, 20 and 30 min, were applied respectively. Day 3 larvae were sampled for triploidy and survival. Percent triploidy was verified using flow cytometry (FCM). Results show that optimal inductions that combine both high rates of triploidy and reasonable survival were those treatments commenced 15 or 20 min post fertilization and continued for 20 or 30 min, using 100 μM 6‐DMAP. These conditions induced rates of triploidy and relative survival of 80.5–93.3% and 36.5–40.2%, respectively, in the first experiment, and corresponding rates were 79.1–93.6% and 20.7–43.0% in the second experiment. High percent triploidy were also obtained in a number of treatments using 150 μM 6‐DMAP, but with overall survival rates generally lower than those using 100 μM 6‐DMAP.     

即食半干虾仁加工技术研究

以凡纳滨对虾（Litopenaeus vannamei）为试验原料,探索半干即食虾仁的加工技术。对蒸煮时间及顺序、调味配方、干制方式及杀菌条件等制作工艺关键步骤进行了研究,制作出风味浓郁、口感良好的半干即食虾仁。最佳的工艺条件为先蒸煮后调味,蒸煮时间为5min,通过正交试验筛选调味配方为1L水需加盐50．0g、糖50．0g、柠檬酸2．0g、味精5．0g,二段干燥,先后于50℃和60cC各烘干1h,80～85℃杀菌处理30rain,此条件下可以较好地保持产品的品质和风味。该研究为对虾的开发利用及提高产品附加值提供了新途径。     

Ⅱ型鲤疱疹病毒ORF121蛋白的多克隆抗体制备及鉴定

针对CyHV-2病毒ORF121基因(GenBank:AFJ20543.1)进行原核表达系统的构建,将纯化重组蛋白作为抗原来免疫BALB/c小鼠获得多克隆抗体,应用该抗体开展CyHV-2病毒诊断及其感染机制研究。以CyHV-2病毒感染细胞上清液为扩增模板,扩增ORF121基因构建至pGEX-4T原核表达载体,经异丙基硫代半乳糖苷(IPTG)诱导表达rORF121重组蛋白,利用尿素纯化后免疫6周龄BALB/c小鼠制备多克隆抗体。结果显示,CyHV-2病毒ORF121基因可在原核表达系统中高效表达目的重组蛋白rORF121,经SDS-PAGE分析大小约为60 ku,主要以不可溶的包涵体存在。利用尿素溶解rORF121蛋白免疫BALB/c小鼠获得抗ORF121蛋白的多克隆抗体,Western Blot实验显示,该抗体可特异性识别CyHV-2病毒感染RyuF-2细胞样品。研究表明,利用CyHV-2感染RyuF-2细胞后,本研究制备的抗ORF121蛋白的多克隆抗体能够通过间接免疫荧光实验特异性识别CyHV-2病毒感染的细胞样品。本研究制备的抗ORF121蛋白的多克隆抗体,能够为CyHV-2病毒诊断技术的构建以及深入开展CyHV-2病毒感染机制提供良好的技术基础。     

Role of free neuromasts in larval feeding of willow shiner Gnathopogon elongatus caerulescens Teleostei, Cyprinidae

ABSTRACT:   It has been reported that the larvae of willow shiner Gnathopogon elongatus caerulescens have many free neuromasts on their body surface. This study examined the ability of the willow shiner larvae to feed on zooplankton by mechanoreception by the free neuromasts. Feeding experiments using untreated larvae and larvae treated with streptomycin, which impairs free neuromast function, were conducted under light and completely dark conditions. The larvae were put into Petri dishes, then nauplii of Artemia salina were introduced. The average number of Artemia eaten by the larvae was expressed as the average ingestion rate of Artemia /10 min. The ingestion rate of Artemia for untreated larvae was 12.3 under light conditions and 10.6 Artemia /10 min. even in complete darkness. The ingestion rate in the larvae treated with streptomycin was 11 under light conditions and only 0.8 Artemia /10 min under dark conditions. The low rate in the treated larvae under dark conditions must be due to impairment by the streptomycin. Therefore, the high ingestion rate for the untreated larvae under dark conditions would be surely dependent on mechanoreception. The results indicate that larval willow shiner is able to feed on zooplankton under limited light conditions.     

Restructured Crispy Fish Cubes Containing Salicornia bigelovii Torr. Developed with Microwave Vacuum Drying

Restructured crispy fish cubes containing Salicornia bigelovii Torr. (S. bigelovii) were developed using microwave vacuum drying. A series of drying experiments and optimization experiments were conducted to optimize the process conditions of microwave vacuum drying. The quality parameters of fish cubes such as crispness, expansion ratio, and sensory score were measured. The optimal expansion ratio and crispness were decided on the basis of optimization experiments by response surface analysis. The results showed that the optimum microwave vacuum conditions were: initial moisture content of surimi was controlled at 60.5% (wet basis, w.b.) and microwave heating for 20 min at the vacuum pressure of 0.090 MPa and microwave power of 11 W/g. The final product had a uniform shape, homogenous texture, pleasant color, and delicious flavor.