紫贻贝和厚壳贻贝杂交及F1代杂交优势初探

2005-2007年对紫贻贝(Mytilus gallo provincialis)和厚壳贻贝(Mytilus coruscus)种间进行了杂交生产性试验,通过亲贝强化培育、确认雌雄亲贝、改革育苗水体交换方法、投喂混合单细胞藻类、流水培养附着稚贝等技术,获得F1代.结果表明,正交F1代幼虫的成活率明显高于反交F1代和厚壳贻贝,在幼虫培育前期与紫贻贝的成活率相当,但随着幼虫的发育有高于紫贻贝的趋势.正交F1代幼虫的壳长和壳高在培育阶段的前期生长与紫贻贝相当,低于厚壳贻贝,高于反交F1代;但后期的生长明显增快,并高于反交F1代和其他两种贻贝.各试验组贻贝海区养殖的成活率在90%以上,正交F1代最高,显著高于反交F1代.各试验组生长特性,正交F1代紫贻贝厚壳贻贝反交F1代.其中正交F1代获附着稚贝3.77×109 ind.正交F1除在孵化率上低于紫贻贝、厚壳贻贝外,在壳长、壳高、成活率等方面指标具有一定杂交优势,而反交F1代的杂交优势不明显.     

厚壳贻贝人工育苗关键技术探讨

正近年来,随着厚壳贻贝养殖规模扩大,厚壳贻贝人工育苗也迅速发展起来。但育苗技术一直没有很好解决,导致育苗数量不稳定,苗种供应不足。本文根据笔者近几年的生产实践经验,对厚壳贻贝的苗种培育过程中关键技术进行总结,以求对今后厚壳贻贝育苗生产有所帮助。一、壳顶幼虫培育壳顶幼虫培育是厚壳贻贝人工育苗过程中的一个重要环节,它决定了苗种生产出苗率高低以及出苗数量。壳顶幼虫培育期间,要做到合理投喂饵料,逐步加大换水量,及时进行疏苗,轮流投洒抗生素,还要勤在显微镜下观察幼体生长发育、活力以及肠胃饱     

脂多糖对细菌生物被膜形成及厚壳贻贝幼虫变态的影响

为探究脂多糖(LPS)对海洋细菌生物被膜形成、海洋贝类幼虫变态所产生的作用,本研究使用不同浓度的LPS直接刺激厚壳贻贝幼虫,观察其对幼虫附着变态的直接作用;同时在海假交替单胞菌形成生物被膜的过程中添加不同浓度的LPS,通过分析生物被膜生物学特性的变化,及变化后的生物被膜对厚壳贻贝幼虫附着变态的影响。研究结果显示,3种浓度的LPS均可直接诱导厚壳贻贝幼虫的变态;10 mg/L浓度LPS处理后的生物被膜其细菌密度、膜厚明显降低(p<0.05),胞外产物中多糖(如可乐酸)、脂类显著增加(p<0.05),同时其对幼虫变态的诱导作用也显著提高(p<0.05),因此推测,在厚壳贻贝幼虫的附着变态过程中,LPS具有直接诱导作用;除此之外,脂多糖还可以通过调控生物被膜胞外物质,特别是可乐酸的生成,从而间接影响厚壳贻贝幼虫的附着变态。     

厚壳贻贝人工繁殖技术的研究

厚壳贻贝(Mytilus coruscus)是我国贻贝的主要养殖品种,其苗种主要依靠天然苗种和半人工采苗获得。2006年浙江海洋学院于嵊泗县石柱育苗厂,首次突破了厚壳贻贝规模化全人工繁殖和稚贝海区中间培育技术。研究结果表明,厚壳贻贝亲贝通过室内强化培养,经人工催产可获得成熟受精卵,受精率可达95%。在水温16℃时,受精卵在受精后25min出现第一极体,受精后39h50min发育至直线绞合幼虫期,胚胎孵化率达92%,在水温15.8～21℃的条件下,经39d室内人工培育,获平均壳长0.694mm的附着稚贝1304.7×104ind;附着稚贝经102d海区中间培育,获平均壳长13.95mm的稚贝404.46×104ind,海区保苗成活率达31%。该研究结果为今后厚壳贻贝大规模苗种生产奠定了重要理论和技术基础。     

培养基对微生物被膜形成和厚壳贻贝附着的影响

为探讨微生物被膜生物学特性、培养基以及大型生物附着三者之间的相互关系,本实验通过微生物分子生态学和海洋贝类生态学等方法调查了不同培养基对海洋细菌所形成微生物被膜的影响及其对厚壳贻贝幼虫附着的影响。结果显示,厚壳贻贝稚贝率与培养基类型和细菌初始密度显著相关,进一步分析表明,Zo Bell 2216E和Seawater Luriabertani(SWLB)条件下分别有6株和9株细菌形成的微生物被膜密度与稚贝率显著相关。扫描电镜结果显示,Staphylococcus sp.3在Zo Bell 2216E培养基条件下形成微生物被膜的细菌分布较为紧密,Pseudoalteromonas sp.8则在SWLB培养基条件下微生物被膜细菌分布较为紧密,且形态变为短杆状。SDS-PAGE结果显示,相比Zo Bell 2216E培养基,Staphylococcus sp.3在SWLB培养基条件下9条条带蛋白显著下降,其中2个条带完全消失;Pseudoalteromonas sp.8则在SWLB培养基条件下明显增加5条蛋白条带。研究表明,微生物被膜的形成受到培养基的影响,培养基的不同导致微生物被膜的形态结构、分布和蛋白有所差异,最终导致微生物被膜诱导厚壳贻贝幼虫附着变态的活性差异,本研究为后续开展厚壳贻贝附着的分子机制奠定良好基础。     

生态因子对厚壳贻贝眼点幼虫生长和存活的影响

为了实现厚壳贻贝人工育苗技术突破和推广,研究了海水盐度、温度、pH等生态因子对厚壳贻贝眼点幼虫的生长和存活的影响,筛选出最佳的繁育条件.结果表明:厚壳贻贝眼点幼虫对盐度的适应范围为23.5～39.1,最适为27.4～35.3;对温度的适应范围为18～27℃,最适为21～24℃;对pH值的适宜范围为7.5～9.0,最适为7.8～8.5.掌握了厚壳贻贝眼点幼虫对主要生态因子的适应范围,对厚壳贻贝人工育苗技术的突破具有重要的指导意义.     

维生素B7和B12对细菌生物被膜形成及厚壳贻贝幼虫变态的影响

为探究B族维生素对海洋细菌生物被膜形成、海洋贝类幼虫变态所产生的作用,本研究首先使用维生素B7 (VB7)和B12 (VB12)直接刺激厚壳贻贝(Mytilus coruscus)幼虫,观察其对变态的直接诱导活性;然后通过添加VB7和VB12,与海假交替单胞菌(Pseudoalteromonas marina)共同形成生物被膜,分析B族维生素对生物被膜形成及其生物学特性的影响;同时检测生物被膜变化对厚壳贻贝幼虫变态发育的影响。研究结果显示,0.02 mmol/L浓度VB7和VB12可以直接诱导厚壳贻贝幼虫的变态,且效果最为显著(P<0.05);0.02 mmol/L浓度VB7和VB12处理后的海假交替单胞菌生物被膜对幼虫附着变态的诱导作用均显著提高(P<0.05);进一步通过细菌密度计数、膜厚度分析、可拉酸染色和定量等方法,揭示VB7和VB12处理后生物被膜细菌密度、膜厚度以及胞外多糖、蛋白和脂质均显著增加(P<0.05)。研究结果证实,VB7和VB12可能通过改变海洋细菌生物被膜的生物学特性,进而调控厚壳贻贝幼虫变态发育。本研究为探究厚壳贻贝幼虫附着变态的分子机制提供了新的理论依据和创新思路,同时为B族维生素在提高厚壳贻贝人工育苗技术、改善厚壳贻贝养殖产业问题和促进海洋牧场生态修复建设等方面的应用提供了理论基础。     

坛紫菜丝状体种质保存技术的研究

采用液体培养基、固体培养基以及胶囊化冷冻法保存坛紫菜丝状体种质，探讨了不同保存条件对其生存的影响。结果表明：液体培养基保存丝状体，在低温5℃，培育30d时，藻丝的增重率为-0．67％；温度为10℃，有利于延长更换培养液的时间，即使120d不换培养液，丝状体也能正常生长。固体培养基保存丝状体，在5℃时固体平板上的细菌生长慢、数量少，但藻丝无法适应低温条件，逐渐死亡；丝状体在10℃固体平板上生长缓慢，但保存100d以上仍然正常生长，达到了长期保存的目的：在胶囊化低温保存中，丝状体存活率较高，而且随着保存时间的延长，成活率没有显著的变化，保持在75％以上。     

墨西哥湾扇贝选育系早期发育与生长

在水温28~29℃、盐度24~25条件下,研究墨西哥湾扇贝(Argopecten irradians concentricus)选育系的早期发育及幼虫和稚贝的生长。受精卵经50 min发育至卵裂,经22~24 h发育至D形幼虫,经11 d发育至眼点幼虫并出现足,进入附着变态阶段。选育系的胚胎发育时间和浮游幼虫前期壳长、壳高与对照系比较差异不显著(P0.05);浮游幼虫后期及至附着变态阶段,选育系与对照系的壳长和壳高差异显著(P0.05)。选育系的附着变态率和存活率与对照系比较均差异显著(P0.05),且优势明显。本研究表明,墨西哥湾扇贝浮游幼虫发育前期缓慢,幼虫后期、稚贝期生长迅速;壳长和壳高的生长速度不同步,幼虫前期壳长生长快于壳高,后期壳高生长快于壳长,并于第28~30天时壳高超过壳长。墨西哥湾扇贝选育系在幼虫和稚贝阶段生长速度明显快于对照系,具有明显的生长优势及生长潜力。     

岐脊加夫蛤幼虫培育密度的初步探讨

本文主要对岐脊加夫蛤幼虫培育的适宜密度进行初步探讨，以壳长为１６０．３μｍ的壳顶幼虫进行实验，历时８天。实验结果表明：在水温２７－２９℃，比重１．０２０－１．０２２的条件下投以等量的异胶藻和牟氏角毛藻的混合饵料，幼虫培育的适宜密度为５－１０个／ｍｌ。密度为２个／ｍｌ幼虫的生长速度快，但变态时存活率较低；     

Larval rearing of calico scallops, Argopecten gibbus, in a flow-through system

A flow-through (FT) culture system is described for calico scallop, Argopecten gibbus, larvae. Its performance was assessed by larval survival rate, shell growth, settlement rate and post-larval shell growth for the duration of larval life (13 days). Comparisons were made with larvae reared in standard static system (S). Effect of increased larval density on FT performance was also investigated. With comparable larval densities, survival rate of Day 2 larvae to pediveliger stage was similar in both larval rearing systems. Shell growth for FT-reared larvae was comparable or significantly higher than in the static system (P < 0.01). Settlement rate of pediveligers was comparable for both systems, averaging 30.7%, and no significant difference was seen in shell growth of FT- and static-reared pediveligers. Increased initial larval density did not affect survival rate in FT, but did negatively affect larval shell growth, settlement rate and post-larval shell growth yielding lower growth and minimal settlement rate (10.9 ± 2.8%) compared to the static system. This FT system was successful as larval rearing system, optimising space allocation in the hatchery, reducing labour, and eliminating the use of antibiotics. Optimising initial larval density within the system needs to be investigated in association with food ration.     

紫贻贝和厚壳贻贝杂交及F1代杂交优势初探

2005年-2007年对紫贻贝和厚壳贻贝种间进行了杂交生产性试验，通过亲贝强化培育、确认雌雄亲贝、改革育苗水体交换方法、投喂混合单细胞藻类、流水培养附着稚贝等技术，获得F₁代。结果表明，各试验组生长特性，正交F₁代﹥紫贻贝﹥厚壳贻贝﹥反交F₁代。其中正交F₁代获附着稚贝3.77亿粒，平均壳长为1.13 mm、平均壳高为0.78 mm，育苗成活率达35.57%。正交F₁除在孵化率上低于紫贻贝、厚壳贻贝外，在壳长、壳高、成活率等方面指标具有一定杂交优势，而反交F1代的杂交优势不明显。     

大竹蛏生产性人工繁育试验

2007年对大竹蛏进行了生产性人工繁育试验。对亲本不同批次催产效果、不同规格亲本产卵差异、孵化密度进行了比较,研究了D形幼虫发育与温度的关系以及幼虫发育各阶段生长规律等,结果显示,大竹蛏属分批成熟、分批产卵的潜居性贝类,不同批次个体平均产卵量(36~118)×104cell,个体总产卵量294×104cell。壳长8~10 cm的亲贝产卵量大,催产效果好。受精卵适宜的孵化密度为5~10 cell/mL。在水温21~24.5℃条件下,D形幼虫经8~5 d生长,发育变态为附着稚贝。D形幼虫壳长日生长33.33±4.22μm,1 mm前的稚贝壳长日生长108.33±22.57μm,1~10 mm的稚贝平均旬生长量2.12 mm,10.4~30.6 mm的苗种平均旬生长量为2.02 mm。当水温下降到15℃后,大竹蛏苗种生长滞缓。2007年培育出壳长11.52±0.27 mm的稚贝2 165×104cfu,平均成活率12.1%。     

肾上腺素对岩牡蛎幼虫变态的诱导

为了得到壳型规则、大小均一的单体牡蛎,本研究采用肾上腺素对岩牡蛎单体苗种生产的诱导条件进行研究,阐明了岩牡蛎单体苗种的最适诱导浓度、诱导时间和诱导密度。结果显示,肾上腺素能显著诱导岩牡蛎幼虫的不固着变态,最适诱导浓度为5×10–5 mol/L,最适诱导时间为1 h,提高诱导浓度和延长诱导时间导致岩牡蛎幼虫的不固着变态率、稚贝壳高和存活率显著降低;肾上腺素对低于8个/mL幼虫密度的诱导效果差异不显著,但稚贝壳高和存活率在8个/mL的培育密度下显著低于0.5～4个/mL,研究表明利用肾上腺素诱导岩牡蛎单体时,可大批量处理眼点幼虫,但稚贝充气培养的最适培养密度应不高于4个/mL。     

Effect of Stocking Density on the Growth and Survival of Spotted Sand Bass Paralabrax maculatofasciatus Larvae in a Closed Recirculating System

Abstract.— The effect on growth and survival of the initial stocking density (50, 100, 150, and 200 larvae/ L) in larval rearing of spotted sand bass was evaluated over 30 d in a closed recirculating system. Larvae were fed with rotifers, copepods, nauplii and adult Artemia , and spotted sand bass yolk-sac larvae. Water quality was monitored daily. The notochordal or standard length of sampled larvae was measured by image analysis. Specific growth rates at each density were compared by covariance analysis. Survival was estimated from day 15 to the end of the experiment, when a resistance test was used to evaluate the juvenile quality among densities. At the end of the experiment, mean standard length of larvae at lower densities was significantly larger ( P < 0.05) than at higher densities. Higher specific growth rates were found at lower densities. Significantly higher survival ( P < 0.05) was recorded for the lowest density, but the highest number of harvested fish was obtained with the highest densities (150 and 200 larvae/L). The lowest density also showed the significantly ( P < 0.05) higher survival after using a resistance test. We conclude the highest density can be used in larval rearing of spotted sand bass. However, better survival, growth, and seed quality are obtained at the lowest density. To recommend an optimal density for this specie, it is necessary to improve the water quality in the culture system and to make a cost-benefit study.     

The effects of dietary lipid sources and lecithin on the production of giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> larvae in the Mekong Delta region of Vietnam

The giant freshwater prawn, Macrobrachium rosenbergii, is cultured widely in the Mekong Delta region of Vietnam but it is often difficult or expensive for hatchery operators to purchase commercial diets used as a feeding supplement to Artemia nauplii. Therefore, in the present study, the effects of lipid sources and lecithin on the growth and survival rate of M. rosenbergii larvae were examined in order to develop suitable hand-prepared larval diets for seed production of M. rosenbergii in this area. Six egg custard diets consisting of various ratios of lipid (originating from soybean oil and squid oil) and lecithin were used for rearing Macrobrachium rosenbergii larvae. Treatments in which larvae were fed diets containing squid oil exhibited the highest body length and survival rates (7.14–7.43 mm and 51.1–68.1%, respectively), and differed significantly from other treatments (P<0.05). Use of dietary soybean oil yielded the lowest body length and survival rates (6.29–6.75 mm and 22.0–48.7%), respectively). The supplementation of dietary lecithin did not increase final body weight but did improve larval survival rates. The n-3 HUFA content of prawns fed dietary squid oil was higher than those of animals provided with other diets. These results indicated that the most appropriate diet for rearing M. rosenbergii larvae is the diet containing 3% squid oil and 1.5% lecithin.     

Experimental culture of larvae,post‐larvae and juveniles of the West Indian top shell,Cittarium pica (Linnaeus 1758)

The West Indian top shell, Cittarium pica, is an endangered vetigastropod of ecological and commercial value from the Caribbean. In order to assess the use of aquaculture as a tool for its sustainable production and conservation, embryos were produced in hatchery and experimentally cultured until juveniles under different conditions. Embryos were incubated under two temperatures (25 and 29°C) and six densities (0.3, 0.6, 1.0, 4.0, 9.0 and 37.0% of bottom‐coverage). Larvae were reared under different temperatures (25 and 27°C), densities (0.5, 1, 1.5 and 10 larvae/ml) and culture systems (static and down‐welling). Post‐larvae were obtained in three different settling conditions, and the early juveniles were cultured supplying fresh seaweed (Laurencia obtusa and Padina gymnospora), natural multi‐specific biofilm and Cylindroteca sp. biofilm. The growth and survival of embryos and larvae were not affected by temperature, but they were affected by density. Higher values were obtained at low densities (<1% of embryos bottom‐coverage and <1 larvae/ml), except for the growth of larvae, which was similar among treatments. Higher larval survival was recorded using the static culture system, and higher percent of post‐larvae recovered was associated with lower density of crawling veliger (0.2 and 0.1/ml), while its higher growth rate was related to the use of biofilms with conspecific mucus and low water‐flow (50%/h). The type of food tested did not affect the juvenile growth, but higher survival was obtained in those fed with seaweed than with multi‐specific biofilm.     

Implications of larval diet concentration on post-larval yield in a production scale flow-through system for scallops (Pecten maximus Lamarck) in Norway

In order to optimize Pecten maximus larval performance and post-larval yield, larvae were fed five algal concentrations in the range 3–20 cells μL^?1 in 2,800-L flow-through tanks without prophylactic antibiotics. Competent larvae were transferred to a commercial hatchery for settlement and provided uniform conditions for 4 weeks to observe effects. Increased diet concentration increased the sum of fatty acids (FA) in the total larval population, reaching 7 and 25 ng FA larvae^?1 at 3 and 16 cells μL^?1, respectively. The FA level in competent larvae was not affected by diet concentration and ranged from 30 to 46 ng larvae^?1. Increased diet concentration increased larval growth rate, and the larvae were ready to settle 5 days earlier when fed 16 cells μL^?1 compared to 3 cells μL^?1. Larval ingestion rate increased during life span and with increased larval diet concentrations, but a considerable amount (40–60 %) of the added algal cells was lost from the larval rearing tanks due to the seawater flow. There was no effect on larval survival, final post-larval shell height, % of competent larvae transferred to settlement, or total yield of post-larvae. Final mean post-larval shell height was 509 μm and 25.5 % of competent larvae settled, resulting in a final post-larval yield between 6.9 and 17.6 % of the initial number of d3 larvae. Competent larvae with similar FA content produced similar numbers of post-larvae independent of diet concentration, even if higher diet concentrations resulted in higher rates of larval development and metamorphosis.