6_二甲基氨基嘌呤诱导太平洋牡蛎三倍体_抑制受精卵第一极体释放

于 1996～ 1997年 ,用 6-二甲基氨基嘌呤 ( 6-DMAP)抑制受精卵第一极体的释放 ,诱导太平洋牡蛎产生三倍体。选用L16( 45)设计 ,进行三因素四水平的正交试验 :6-DMAP浓度 ,设15 0、30 0、45 0和 60 0μmol/L ;诱导时机 (即精卵混合后的时间 ) ,设 10、15、2 0和 2 5min ;诱导持续时间 ,设 10、15、2 0和 2 5min。试验平行重复二次。结果为最高三倍体诱导率为 ( 71.3± 1.2 ) % ,该实验组胚胎孵化率为 ( 5 5 .5± 3.1) % ,D形幼虫畸形率为 ( 10 .7± 1.6) %。根据直观分析结果 ,得出诱导太平洋牡蛎三倍体各因素的最优水平组合 :水温 2 5～ 2 5 .5℃ ,精卵混合 10min时 ,将受精卵浸泡在含 6-DMAP 60 0μmol/L的海水中 15min ;决定三倍体产生的三因素的主次顺序 :6-DMAP浓度→诱导时机→诱导持续时间。 6-DMAP浓度对三倍体诱导率影响显著 ,但诱导时机和诱导持续时间不显著。     

杂交鲍三倍体诱导技术研究

为研究杂交鲍的三倍体诱导技术,通过预试验,观察和了解23.5 ℃下杂交鲍受精卵第一极体和第二极体释放的时间。采用细胞松弛素B抑制第二极体释放的方法,分别在不同的诱导起始时间、诱导剂浓度及诱导持续时间下抑制第二极体的释放,测定鲍的三倍体率,进行了杂交鲍三倍体的诱导试验。试验结果表明:杂交鲍在水温23.5 ℃下,处理起始时间为20 min,诱导剂浓度为0.70 mg·L-1,诱导的持续时间为10 min时,表观三倍体率最高,达75 %以上。检查稚鲍的倍性,三倍体率达到66 %。     

6-DMAP诱导栉孔扇贝异源雌核发育二倍体

采用紫外线遗传灭活的长牡蛎精子激活栉孔扇贝卵子，运用L9（3^4）设计不同6-DMAP的处理条件来诱导染色体加倍，获得第二极体抑制型栉孔扇贝雌核发育二倍体早期胚胎。以雌核发育担轮幼虫为材料，滴片法制备染色体并采用计数法统计倍性，分析不同6-DMAP浓度、诱导时机和诱导持续时间对雌核发育二倍体诱导率的影响，并统计早期胚胎畸形率。结果表明，不同6-DMAP浓度、诱导时机和诱导持续时间对异源精子诱导栉孔扇贝雌核发育二倍体的诱导率均有显著影响；早期胚胎畸形率与二倍体的诱导率呈负相关，y=46.632-0.891x（r=-0.813，P〈0.01）。6-DMAP诱导栉孔扇贝雌核发育二倍体的最佳处理条件为：20%～25%受精卵排放出第1极体时，60μg/ml的6-DMAP持续处理25min可得到最高的诱导率为29.5±5.36%。研究结果为6-DMAP诱导栉孔扇贝雌核发育染色体加倍提供了一定的细胞学依据和技术参数。     

6-二甲基氨基嘌呤诱导泥蚶三倍体实验

于2001年5月,用6-DMAP抑制泥蚶受精卵第二极体的释放诱导三倍体的产生。实验设三个6-DMAP浓度梯度,分别为300μmol/L、450μmol/L、600μmol/L,处理时机为受精卵第一极排放体30％,处理持续时间为10min。结果:三倍体诱导率300μmol/L时为35.5％,450μmol/L时为48.6％,600μmol/L时约为58.5％,另外本实验还就6-DMAP浓度对D形幼虫孵化率和畸形率的影响以及在染色体制片过程中的有关问题进行了初步探讨。     

6-DMAP诱导扇贝异源三倍体的细胞学观察

分别在受精后15和30mjn,用60 mg·L~(-1)6-二甲基氨基嘌呤(6-DMAP)处理栉孔扇贝(♀)×虾夷扇贝(♂)受精卵诱导异源三倍体,持续处理5～25min。采用二氨基苯基吲哚(DAPI)染色和荧光显微镜观察对处理前后受精卵的染色体行为变化进行了研究,空气干燥法制备染色体检测胚胎倍性。结果表明,无论抑制第一极体或是第二极体释放均能获得正常发育的异源三倍体胚胎;抑制第一极体释放,产生异源三倍体、非整倍体,延长处理时间能获得五倍体;抑制第二极体释放,产生异源三倍体;6-DMAP使染色体运动终止,处理前后,核相无变化,随着处理时间的延长,核相泡状化增强。     

不同方法制备的三倍体长牡蛎养殖效果的比较

比较了细胞松弛素B(CB)和6-二甲基氨基嘌呤(6-DMAP)通过抑制受精卵极体释放的方法批量诱导三倍体长牡蛎的养殖效果.长牡蛎卵子在25℃的海水中受精,20～30min后,开始用浓度为0.5mg@L-1的CB处理,持续18～22min,受精卵处理密度为4.0～4.5×107个@L-1,三倍体产率为65.2%～70.1%,面盘幼虫孵化率为12.3%～14.5%,诱导效率指数为0.09.6-DMAP的使用浓度为400～420μmol@L-1,受精卵处理密度为3.0～3.5×107个@L-1,授精水温、处理起始和持续时间等与CB方法相同,三倍体产率为58.7%～65.4%,面盘幼虫孵化率为52.1%～55.4%,诱导效率指数为0.32.两种方法的采苗率基本相同,采苗器为基质较硬的栉孔扇贝贝壳.海区养殖采用浮筏夹苗吊养技术,两种方法诱导的三倍体牡蛎养殖性状没有明显差别.通过比较CB和6-DMAP两种诱导方法及三倍体的养殖效果表明,后者具有更好的应用性.     

CB与6-DMAP诱导香港牡蛎三倍体的效果比较

以三倍体率、卵裂率、D幼率、生产成本等为指标,比较了CB、6-DMAP两种化学试剂诱导香港牡蛎三倍体的效果,研究了试剂浓度、诱导时机、诱导持续时间及受精卵密度等4种因素对香港牡蛎三倍体的诱导效应。结果显示,在温度28~30°C、盐度15~25,受精卵密度为2.0×10~8个/L条件下,采用0.5 mg/L的CB在受精后15~18 min处理,诱导持续时间为20 min,可产生100%三倍体;合子的卵裂率为53.16%~63.00%,D形幼虫孵化率为47.32%~53.09%,诱导效率指数为0.47~0.53,生产成本为260元/L。相同条件下,采用浓度为75~100 mg/L的6-DMAP处理,诱导持续时间为20~25 min,受精卵处理密度4.5×10~7个/L,可产生62.52%~72.36%的三倍体;合子的卵裂率为60.00%~66.25%,D形幼虫孵化率为74.43%~90.00%,诱导效率指数为0.47~0.65,生产成本为139~185元/L。综合比较两种方法,6-DMAP诱导方法更加适合用于大规模的香港牡蛎三倍体苗种生产。本研究为香港牡蛎多倍体育种提供了研究数据与实践经验。     

一种生态化鲍育苗和养成模式的构建

该研究目的是建立一套鲍育苗和养殖的生态化模式,包括鲍苗种生态培育模式、鲍陆基生态养成模式和海上鲍藻套养生态养成模式,以克服目前鲍养殖业存在的病害频发、成活率低下等问题,提高鲍养殖的经济效益,降低养殖风险。鲍苗种生态培育模式通过＂光照-微藻-微生态制剂-滤食性动物-鲍苗＂构建复合生物体系;鲍陆基生态养成模式通过池内建立＂复合生物区系＂,并在池外设立生态塘和生态渠进行养殖废水处理的方式构建;海上生态养成模式通过＂鲍藻套养＂构建。共培育鲍苗5批次,培育周期210～240d,共产出鲍苗17.764×104个,平均壳长1.5cm以上,育成率0.78%～1.23%。陆基生态化模式养成商品鲍43.774×104个,平均壳长6.53cm,平均体质量34.6g,成活率87.5%,周期18～20个月。海上鲍生态养成150口网箱,养成商品鲍3×108个,平均壳长6.50cm以上,成活率85%以上,养成时间18～20个月。     

低渗诱导太平洋牡蛎三倍体以及与其他诱导方法的比较

通过低渗方法抑制受精卵第二极体(PB2)的释放,诱导太平洋牡蛎(Crassostrea gigas)三倍体.在水温为23～25℃的条件下,分别采用不同低渗海水(盐度分别为4、6、8、10、12、14、16),在不同处理时机,即第1个第一极体(PB1)出现、30%的PB1出现、40%的PB1出现、50%的PB1出现以及第1个第二极体(PB2)出现时,对太平洋牡蛎受精卵进行10 min、15 min、20 min、25 min持续处理.处理后,于正常盐度海水中孵化,收集D形幼虫,通过流式细胞仪进行倍性测定,确定最佳诱导条件.结果表明,当太平洋牡蛎受精卵PB1出现40%时,在盐度为8的低渗海水中,持续处理15 min,得到最高三倍体诱导率为(89.16±1.39)%.与对照组相比,低渗诱导获得的三倍体幼虫表现出明显的生长优势.将低渗方法同高温(32℃)、低温(2℃)和6-DMAP(450μmo1/L)处理等多倍体诱导方法进行比较,结果显示,低渗组的卵裂率和孵化率显著高于高温、低温和6-DMAP诱导的太平洋牡蛎三倍体(P＜0.05).6-DMAP诱导的三倍体率虽略高于低渗诱导,但差异不显著(P＞0.05).通过综合评价指数(Ie)比较,认为低渗诱导多倍体的方法比高温、低温和6-DMAP诱导具有明显的优势.     

山东南部皱纹盘鲍工厂化养殖试验研究

报道了山东南部皱纹盘鲍（HaliotisdiscushannaiIno）工厂化养殖试验研究的结果。自1995年1月19日至1996年5月19日，经16个月的培育，将平均壳长21．4mm，平均体重0．99g的幼的培育成平均壳长61．9mm（最大壳长72．0mm），平均体重28.5g的成鲍，成活率为93．2％。0．3m2的波纹权的养殖密度：鲍壳长为20．0～30．0mm时，养殖密度200～250粒为好；40．0mm以前，养殖密度150粒为宜；60．0mm以前，养殖密度50～80粒为佳。饵料以人工配合饵料为好。在鲍壳长40．0mm以前，在投喂人工配合饵料的同时，加上适量的新鲜海藻可以达到同样的增长和增重的目的。全投喂海藻增长和增重都较慢。     

Growth and feeding in juvenile triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814), at two temperatures

Growth and feeding of juvenile triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were investigated at two temperatures of 17 and 21 °C over a 50‐day period. There were no differences in growth between triploid and diploid abalone as measured by shell length and body weight. Both triploid and diploid abalone increased in length but not in weight at 21 °C. Condition indices were similar for triploid abalone maintained at both temperatures; however, those for diploid abalone were significantly higher at 17 °C than at 21 °C. Food intake was significantly greater yet feed conversion efficiency was significantly lower in triploid than in diploid abalone. Both the feeding variables were independent of temperature. On average, diploid abalone were able to convert 1 g of dry food ingested to 0.58 g of body weight, but triploid abalone only 0.44 g.     

Growth and reproductive performance of triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814)

Growth and reproduction of triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were compared in a 30-month study. Triploidy was induced by inhibition of the second polar body formation using 6-dimethylaminopurine (6-DMAP) or cytochalasin B (CB). There were no significant differences in growth and survivorship between triploid and diploid abalone. However, triploid abalone had a more elongated shell and greater foot muscles than diploid abalone. A slightly curvilinear growth in shell length was conformed to all treatments. While diploid abalone had reached sexual maturity and spawned during the study, gonadal development and gamete maturation were abnormal in triploids. Female triploids lacked an apparent gonad at the macroscopic level but microscopic examination revealed that they had a thin layer of oogonia development. In contrast, male triploids were able to form similar-sized gonads to diploids during most of the reproductive period, but with brown-yellow discolouration and stalled gametogenesis at spermatocyte formation. Sex ratio of triploid abalone did not deviate from 1:1. With the onset of sexual maturation, growth and gonadal maturation occurred concurrently in diploid abalone, and there was no indication that growth of (diploid) abalone was reduced.     

Ability of salmon growth hormone to accelerate somatic growth of juvenil abalone Haliotis discus hannai

ABSTRACT:   The ability of salmon growth hormone (sGH) to accelerate the somatic growth of juvenile abalone Haliotis discus hannai by immersion was examined. After immersion for one hour into a sGH-rich solution at a concentration of 30 mg/L, the sGH immunoreactivity in the body fluid of abalone was maximal after one day, and levels were still detectable at two days. No immunoreactivity was observed in the control group. Following immersion for one hour at one or two-week intervals for 120 weeks into the sGH-rich solution at 30 mg/L, the sGH-immersed abalone exhibited a significant increase in shell length and body weight. On the other hand, abalone immersed into a salmon prolactin (sPRL)-rich solution at 30 mg/L for one hour showed a lower increase in both shell length and body weight than the sGH-immersed abalone. No increases in shell length and body weight were observed in either the bovine serum albumin-immersed abalone or untreated controls. These results provide evidence that sGH can be transported from ambient water into the circulatory system of abalone, and can subsequently improve the somatic growth of juveniles.     

湾外海域不同结构鲍匍匐基初步对比试验

2016年3月6日—5月3日,在福建省漳浦县霞美镇浮头湾外海域进行三种不同结构鲍匍匐基养殖对比试验。三种鲍匍匐基分别为扣箱式(有效匍匐面积1.072 8 m2)、双层斜板式(有效匍匐面积1.993 5 m2)和传统养殖盆(有效匍匐面积0.413 3 m2),试验对象为皱纹盘鲍杂交鲍(Haliotis discus hanai)。试验结果如下:1)扣箱式鲍壳长日增长为0.131mm,日增重为0.129 g,存活率为98.44%,饲料转化率为3.38%;2)双层斜板式鲍壳长日增长为0.098 mm,日增重为0.096 g,存活率为99.38%,饲料转化率为4.29%;3)传统养殖盆式鲍壳长日增长为0.115 mm,日增重为0.103 g,存活率为98.00%,饲料转化率为5.37%。试验设施未受到湾外海况影响破坏。结果表明扣箱式鲍匍匐基整体性能较好,基于该匍匐基的鲍养殖网箱在经济型吊杆辅助作业下具有养殖效率高、抗风浪流性好、劳动强度低、所需劳动力少等优点,适合应用于湾外养殖。     

不同养殖条件下黄缘闭壳龟二龄幼龟的生长研究

研究了常温养殖(气温为14.7～33.3℃)与初夏、秋末阶段性的控温养殖(气温为20.3 ～33.3℃)两种养殖方式下黄缘闭壳龟Cuora flavomarginata(Gray)二龄幼龟生长的情况,试验结果:通过180 d的试验,常温养殖幼龟平均增重54.27 g,体重相对增长率为115.89％,绝对增长率为0.30...     

Effect of three triploidy induction methods on the growth and survival of larvae and post‐larvae of the Caribbean scallop Argopecten nucleus

Argopecten nucleus is a small scallop from the Caribbean Sea and a relatively new species for aquaculture. One of the key challenges to develop the farming operations for this species from the current pilot scale to commercial level is to improve its harvest size. In this study, we tested three different methods for triploidy induction. Additionally, the effect of these protocols on survival, developmental rate and size of larvae and post‐larvae were assessed. Three different mechanisms to stimulate the inhibition of the release of the second polar body were tested; (1) cold shock (18°C); (2) 6‐dimethylaminopurine (6‐DMAP); (3) cytochalasin B (CB) and (4) dimethylsulphoxide (DMSO). The treatment with 6‐DMAP yielded the highest percentage of triploid larvae (39%). The survival and development rate, however, were higher in non‐treated larvae (control) than in the treatment groups. Interestingly, larvae from CB and the DMSO control groups exhibited lower growth rates in length than those from control and the other two treatments. No influence of the triploidy induction treatments was observed on post‐larvae survival, but the size of post‐larvae was larger for the cold shock treatment and DMSO control group. Our results indicate that the use of 6‐DMAP has the greatest potential to produce triploid larvae of A. nucleus without affecting negatively growth and survival of post‐larvae.     

文蛤红壳色选育系F_3的生长规律及模型

为检验文蛤(Meretrix meretrix)红壳色选育系F_3的生长性能,连续比较了同等养殖条件下选育系F_3和自然对照组CG的生长情况,并采用Von Bertalanffy、Gompertz、Logistic和Brody 4种非线性模型对文蛤红壳色选育系F_3的生长参数进行生长模型构建,以研究文蛤红壳色选育系F_3的生长规律。结果表明,文蛤红壳色选育系F_3在浮游期、稚贝中间培育期和塘口养殖期内的生长均显著快于对照组CG(P0.05),F_3的平均日生长率均大于CG,490日龄内F_3的壳长相对于CG平均增长率为34.05%。塘口养殖期间,6~9月是F_3生长最为快速阶段,且F_3较CG的粒重增长率在98.27%以上。文蛤红壳色选育系F_3的壳长和粒重之间遵循复合曲线Y=9.028×1.499W,R2=0.977。文蛤红壳色选育系F_3的壳长生长遵循Logistic生长模型,其在575日龄时出现快速生长,这为及时疏苗养殖提供了依据。     

Feasibility of Offshore Co‐culture of Abalone,Haliotis discus hannai Ino,and Sea Cucumber,Apostichopus japonicus,in a Temperate Zone

Feasibility of offshore co‐culture of abalone and sea cucumber was investigated in Northern China. Survival and growth of abalone, Haliotis discus hannai Ino, and sea cucumber, Apostichopus japonicus, co‐cultured in abalone cages from suspended longlines, in the offshore area, were examined. Abalone and sea cucumbers were co‐cultured at density ratios of both 3:1 and 6:1 for 1 yr. Abalone were fed with fresh kelp and no additional feed was given to sea cucumbers. Survival of abalone and sea cucumber was 100% for all treatments. Abalone and sea cucumber grew well; the body weight (BW) of abalone and sea cucumber was nearly doubled and had reached a commercial size. There were no significant differences in the growth rates for both abalone and sea cucumber between the two density treatments. The specific growth rate of BW of abalone (SGRbw) was highest in June, with a value of 0.536%/d. Growth rate of sea cucumber (SGRsc) was highest in December, reached 1.84%/d, with an annual average SGRsc of 0.182%/d. Results suggested that the offshore co‐culture of abalone and sea cucumber was feasible offshore. The co‐culture of abalone with sea cucumbers may provide an additional valuable crop without additional financial input.     

Effects of alternate starvation and refeeding cycles on food consumption and compensatory growth of abalone, Haliotis asinina (Linnaeus)

The effects of alternate starvation and refeeding on food consumption and compensatory growth of hatchery‐bred abalone, Haliotis asinina (Linnaeus), were determined. Two groups of abalone juveniles (mean shell length = 29 mm, body weight = 5 g) were alternately starved and refed a macro‐alga, Gracilariopsis bailinae at equal duration (5/5 or 10/10) over 140 days. A control group (FR) was fed the seaweed ad libitum throughout a 200‐day experimental period. Starved and refed abalone showed slower growth rates (DGR, 63 and 70 mg/day in the 5/5 and 10/10 groups respectively), as a result of reduced food intake (DFI 15% and 16% day^?1 respectively), after repeated starvation and refeeding cycles. Percentage weight gains (5/5 = 196%, 10/10 = 177%) were significantly lower than that of the control (397%). When refed continuously over 60 days, the starved groups exhibited increased DFI and fed at the rate of 24% and 25% day^?1, which were not significantly different from that of the control at 26% day^?1. At the end of the experiment, no significant differences were observed among three treatments in terms of shell length (range: 46–48 mm), body weight (range 25–28 g), % weight gain (392–465%) and per cent survival (range 87–98%). The results indicated that H. asinina had a complete compensatory growth following a return to full rations after a series of intermittent starvation and refeeding cycles.