华贵栉孔扇贝♀×栉孔扇贝♂远缘杂交的受精细胞学观察

用HOECHST33258对华贵栉孔扇贝(Chlamys nobilis)♀×栉孔扇贝(Chlamys farreri)♂的受精卵进行染色,在荧光显微镜下观察其受精细胞学过程。观察表明:栉孔扇贝的精子能够使华贵栉孔扇贝卵子受精。精子入卵后呈一圆形亮点,体积稍有膨大成球形;授精后成熟卵母细胞释放出第一极体和第二极体后,精核解凝、稀疏、泡状化,形成雄性原核(male pronucleus);雌性原核(female pronucleus)在完成两次成熟分裂之后,染色质去浓缩,扩散膨大;雌雄原核相互靠拢,当雌雄原核膨胀到最大程度时,发生融合,形成合子。受精卵能够正常发育,完成第一次卵裂,卵裂时细胞核分离未观察到异常现象。与华贵栉孔扇贝和栉孔扇贝种内交配对照组相比,异源受精卵的细胞学过程明显滞后,同时其发生过程具明显的不同步现象。实验中还观察到少数的雌核发育现象。     

不同饵料和饥饿对魁蚶幼虫生长和存活的影响

在水温23.2-24.0℃,盐度29.5-30.0条件下,研究了5种单胞藻饵料和饥饿对魁蚶浮游幼虫生长与存活的影响.投喂不同饵料试验中,球等鞭金藻与4种单胞藻混合投喂试验组幼虫的特定生长率、眼点幼虫比例和壳长均显著高于单一投喂试验组,其中球等鞭金藻与小球藻混合投喂的效果最好;投喂单一饵料试验中,球等鞭金藻组幼虫的生长、眼点幼虫比例与存活率显著高于其它试验组.在饥饿试验中,不同饥饿天数条件下幼虫的存活率差异不显著;生长方面,饥饿1-2d较短时间与一直投喂对照组的幼虫相比较,特定生长率、眼点幼虫比例、壳长的差异不显著,但随着饥饿时间延长,幼虫的生长变慢且眼点幼虫比例下降,一直饥饿情况下幼虫的生长基本停滞.研究结果可以为魁蚶人工苗种繁育技术的改进提供参考依据.     

饥饿和再投喂对青蛤(Cyclina sinensis)幼虫生长、存活及变态的影响

在温度18.2～20.6℃,盐度23～25,pH 7.96～8.14 的条件下,研究了饥饿和再投喂对青蛤幼虫生长、存活及变态的影响.结果表明:在饥饿状态下,幼虫具有生长现象,且随着饥饿时间的延长,壳长逐渐接近一个常值而不再生长;幼虫可以由面盘幼虫发育到足面盘幼虫.随饥饿时间延长存活率下降;且足面盘幼虫及其变态规格、单水管稚贝规格随着饥饿时间延长而减小;幼虫的不可逆点(PNR)为12.48d;延迟变态时间长达12.7d.饥饿后再投喂相同的时间,幼虫能够恢复生长,存活的幼虫能够变态;稚贝表现出补偿生长现象,以壳长作为衡量标准,完全补偿生长能力依次为:S10＞S11＞S12＞S1＞S2＞S3;超补偿生长能力依次为:S9＞S8＞S7＞S6＞S5＞S4.     

华贵栉孔扇贝Akirin 2基因的克隆与表达分析

为了解Akirin在华贵栉孔扇贝(Chlamys nobilis)中的作用,本研究分离了华贵栉孔扇贝Akirin 2基因(命名为CnAkirin 2),并描述了CnAkirin 2的特征。使用转录组分析和PCR方法获得了CnAkirin 2 cDNA序列,通过序列比对分析了Akirin氨基酸序列在不同物种中的保守性,使用MEGA 7.0软件的邻接法构建了Akirin 2的系统进化树,使用实时定量PCR分析了Akirin 2在华贵栉孔扇贝6种不同组织中的表达。结果显示:该基因序列全长1 888 bp,开放阅读框长597 bp,编码氨基酸198个;CnAkirin 2的预测分子量为22.11 kD,理论等电点pI为9.30,具有核定位信号PKRRRCM;不同物种Akirin氨基酸多序列比对结果显示,推译的CnAkirin 2氨基酸序列与其他物种Akirin氨基酸序列在N端和C端具有高度相似性;使用邻接法构建的系统进化树显示CnAkirin 2与牡蛎和其它扇贝等贝类Akirin聚为一支;实时定量PCR结果表明CnAkirin 2在不同组织中均有表达,其中在精巢表达量最高,其次为卵巢,在其它组织中的表达水平最低,推测其可能在性腺发育过程中发挥重要作用。本研究为探索Cn Akirin 2在华贵栉孔扇贝中的作用奠定了基础。     

模拟增温下门源草原毛虫幼虫生长发育特征

采用开顶式生长室(OTC)模拟增温的方法研究温度升高对门源草原毛虫幼虫发育历期、生长发育速率、体重及存活等的影响。研究发现:在增温0—1.26℃内,随着增温幅度变大,1龄幼虫越冬后开始活动期和蛹期提前,整个幼虫的发育历期缩短;增温可以提高门源草原毛虫幼虫的相对生长速度,使其发育高峰期提前到5月份;随着增温幅度变大,门源草原毛虫幼虫的体重有减轻的趋势;0—1.26℃的温度升高导致门源草原毛虫幼虫存活率增提高,但是增温过高(1.98℃)也会导致其存活率急剧下降。     

一株形态特殊的细菌对栉孔扇贝致病性的初步研究

为研究分离到的一株形态特殊的细菌对栉孔扇贝(Chlamys farreri)致病性,采用测定病原液蛋白含量的方法,对病原相对定量。用含200～300μg／ml蛋白的病原液进行了不同浓度梯度、同一温度和相同浓度梯度、不同温度对栉孔扇贝的致病作用的测试,结果表明：扇贝人工感染后潜伏期3～7d,死亡高峰期为5～10d,符合一般病原感染的规律。病死贝的一般病理变化与养殖海区自然发病扇贝的病理变化一致。23、26℃下该病原体对栉孔扇贝具有强致病作用。可以确定该细菌为造成栉孔扇贝大规模发病死亡的主要病原之一。     

冬季饥饿再投喂对菲律宾蛤仔生长、存活和生化组成的影响

2007年12月—2008年4月,在冬季低温（水温3.0 ℃～4.2 ℃、盐度25～27、pH 7.90～8.16）条件下,采用饥饿时间不等、恢复投喂时间相同的方式,研究了饥饿再投喂对不同规格菲律宾蛤仔生长、存活及体组分的影响.结果表明：在冬季低温条件下,各试验组菲律宾蛤仔生长性状的差异不明显;其对饥饿的耐受能力随着蛤仔规格的增大而升高;存活率随着饥饿时间的延长而降低,但整个试验过程中未出现不可逆点.从较大规格菲律宾蛤仔（壳长7 mm）的体组分上看：菲律宾蛤仔水分、脂肪含量随着饥饿时间的延长而下降;蛋白质含量在饥饿前期随着脂肪的消耗而上升,后期因脂肪的过度消耗开始被利用而下降;灰分含量在整个试验过程中的变化不显著.     

栉孔扇贝×虾夷扇贝杂交子一代与双亲染色体核型的分析

连续几年栉孔扇贝大面积死亡现象严重制约了北方贝类养殖业的发展 ,造成了巨大的经济损失。栉孔扇贝和虾夷扇贝生物学性状差异较大 ,可望通过杂交途径培育出抗逆性强、生长快的扇贝养殖新品种。杨爱国等[1] 和周丽青等[2 ] 的研究结果表明 ,栉孔扇贝与虾夷扇贝正反交杂交子一代     

温度和盐度对缢蛏浮游幼虫发育的影响

本文叙述了在室内控制条件下,温度和盐度单因子及其结合对缢蛏浮游幼虫的影响。缢蛏浮游幼虫生长发育的适温范围为17°—32℃,其中以25°—27℃为最好。对高温和低温有较强的耐力,致死温度的上限为40℃左右,变态的下限水温为15℃左右,幼虫适盐范围是4.50—28.30‰,其中以12.40‰为最适宜,但幼虫对低盐度的耐力比对较高盐度的耐力强。 温度对缢蛏浮游幼虫期的长短、增长速度和存活率的影响,要比盐度对它的影响显著,而盐度对缢蛏幼虫变态率的影响却比温度重要些。温度与盐度对缢蛏浮游幼虫发育影响的相关性不明显。     

温度和盐度对缢蛏浮游幼虫发育的影响

本文叙述了在室内控制条件下,温度和盐度单因子及其结合对缢蛏浮游幼虫的影响。缢蛏浮游幼虫生长发育的适温范围为17°—32℃,其中以25°—27℃为最好。对高温和低温有较强的耐力,致死温度的上限为40℃左右,变态的下限水温为15℃左右,幼虫适盐范围是4.50—28.30‰,其中以12.40‰为最适宜,但幼虫对低盐度的耐力比对较高盐度的耐力强。 温度对缢蛏浮游幼虫期的长短、增长速度和存活率的影响,要比盐度对它的影响显著,而盐度对缢蛏幼虫变态率的影响却比温度重要些。温度与盐度对缢蛏浮游幼虫发育影响的相关性不明显。     

二种改进人工饲料对红脉穗螟生长发育和繁殖的影响

在温度(29±1)℃和相对湿度75%±5%条件下,研究了2种改进人工饲料和天然饲料椰子幼果对红脉穗螟Tirathaba rufivena Walker生长发育、繁殖力和营养的影响。结果表明:与天然饲料相比,用改进人工饲料饲养的红脉穗螟幼虫生长发育速度、幼虫存活率、蛹重及产卵量均提高;幼虫相对生长率(RGR)、食物利用率(ECI)及食物转化率(ECD)均显著提高,而相对取食量(RCR)则显著降低。表明红脉穗螟幼虫对2种人工饲料有很好的适应性,可用于大量饲养。     

Starvation resistance of gypsy moth,Lymantria dispar (L.) (Lepidoptera: Lymantriidae): tradeoffs among growth,body size,and survival

Summary Survival and body composition of starving gypsy moth larvae initially reared on aspen foliage or artificial diet differeing in nitrogen (N) and carbohydrate concentration were examined under laboratory conditions. Diet nitrogen concentration strongly affected starvation resistance and body composition, but diet carbohydrate content had no effects on these. Within any single diet treatment, greater body mass afforded greater resistance to starvation. However, starving larvae reared on 1.5% N diet survived nearly three days longer than larvae reared on 3.5% N diet. Larvae reared on artificial diet survived longer than larvae reared on aspen. Differences in survival of larvae reared on artificial diet with low and high nitrogen concentrations could not be attributed to variation in respiration rates, but were associated with differences in body composition. Although percentage lipid in larvae was unaffected by diet nitrogen concentration, larvae reared on 1.5% N diet had a higher percentage carbohydrate and lower percentage protein in their bodies prior to starvation than larvae reared on 3.5% N diet. Hence, larger energy reserves of larvae reared on low nitrogen diet may have contributed to their greater starvation resistance. Whereas survival under food stress was lower for larvae reared on high N diets, growth rates and pupal weights were higher, suggesting a tradeoff between rapid growth and survival. Larger body size does not necessarily reflect larger energy reserves, and, in fact, larger body size accured via greater protein accumulation may be at the expense of energy reserves. Large, fast-growing larvae may be more fit when food is abundant, but this advantage may be severely diminished under food stress. The potential ecological and evolutionary implications of a growth/survival tradeoff are discussed.     

幼虫期短时高温暴露对二点委夜蛾存活和繁殖的影响

【目的】随着全球气候变暖,夏季短时极端高温发生的频率逐渐增加。本研究旨在探明二点委夜蛾Athetis lepigone幼虫期对高温的适应性。【方法】将二点委夜蛾不同日龄(1,6,12和18日龄)幼虫在不同高温(35,38和41℃)条件下暴露不同时间(0.5,1,2,4和6 h)后转移至适温(26℃)继续饲养,观察短时高温对其存活率、发育历期、化蛹率、羽化率、雌虫寿命、单雌产卵量及次代卵孵化率的影响。【结果】幼虫期短时高温暴露的温度和时间对二点委夜蛾幼虫的存活率和发育历期有显著影响,而对化蛹率、成虫羽化率、雌虫寿命、单雌产卵量以及次代卵孵化率影响不显著。随着温度的升高和处理时间的延长,幼虫存活率逐渐降低,发育历期逐渐延长。其中,18日龄的幼虫最为敏感,38℃和41℃暴露6 h后存活率分别为58.3%和17.7%,显著低于对照,发育历期分别为25.5 d和29.2 d,较对照显著延长。【结论】幼虫期经历短时高温暴露仅对幼虫的存活和发育历期有影响,而对后续蛹和成虫的生长发育及成虫繁殖力没有影响。     

Effects of food concentration and availability on the incidence of cloning in planktotrophic larvae of the sea star Pisaster ochraceus

A decade ago, cloning was first observed in the planktotrophic larvae of sea stars obtained from plankton tows. However, no controlled experimental studies have investigated what factors may regulate this remarkable phenomenon. In the present study we offer the first documentation of cloning in the planktotrophic larvae of Pisaster ochraceus from the northern Pacific coast. This species was used as a model system to investigate three factors that may influence the incidence of asexual reproduction (cloning) in planktotrophic sea star larvae. In an initial experiment, larvae were reared under nine combinations of three temperatures and three food (phytoplankton) concentrations. Larvae reared at 12-15 degrees C and fed the highest food concentrations grew larger than the other larvae and produced significantly more clones. In a second experiment, qualitatively different algal diets were fed to larvae reared under the conditions found to be optimal in the initial experiment. Up to 24% of the larvae consuming a mixed phytoplankton diet of Isochrysis galbana, Chaetocerous calcitrans, and Dunaliella tertiolecta cloned, and significantly more clones were produced by these larvae than by those fed monospecific diets. Our experiments indicate that cloning generally occurs after larvae have attained asymptotic body length and only when food is abundant and of high quality. Since larval mortality is considered to be extremely high for marine invertebrates with planktotrophic larvae, production of clones under optimal conditions of temperature and food may serve to increase larval populations when the environment is most conducive to larval growth.     

The impact of food type, temperature and starvation on larval development of Balanus amphitrite Darwin (Cirripedia: Thoracica)

The impact of diatom food species (Chaetoceros calcitrans and Skeletonema costatum), temperature and starvation on the larval development of Balanus amphitrite was evaluated. Starvation threshold levels for different ages of larvae (0- to 5-day-old) fed with C. calcitrans and S. costatum and then starved at 5, 15 and 25 °C temperature were estimated as ultimate recovery hour (URH; denoting the starvation point in hours at the end of which larvae can recover and continue development). Effect of temperature on starvation threshold varied significantly with larval age and food species. The URH declined with larval age at 5 °C, but not at 15 and 25 °C. The URH and grazing rates were high for early instars fed on C. calcitrans, and for advanced instars fed on S. costatum. Carbon gain through feeding was maximum for 2-day-old larvae when fed with C. calcitrans and decreased with larval age. However, when fed with S. costatum carbon gain increased with larval age. This confirms that with development the utility of food types changes. The differences in the carbon gain can be attributed to differences in grazing rate due to variations in the size of the diatom cells, larval intersetular distance, diatom sinking rate and the photo-taxic behavior of larvae. Molting was observed at times when larvae were undergoing starvation and this could be viewed as stress-induced molting, and it differed with the larval age and food organisms.     

Effects of starvation on the formation of daily growth increments in the otoliths of milkfish, Chanos chanos (Forsskål), larvae

The effects of starvation on daily growth and increment formation in the otolith were examined using a double oxytetracycline-labelling method on larval milkfish, Chanos chanos (Forsskål), reared under different feeding regimes. The results indicated that the differences in body and otolith growth between the larvae fed once and three times a day were not significant, and that the otolith growth increment was deposited daily in both groups of fed larvae. In contrast, the starved larvae grew at a slower rate than fed larvae in body length and otolith dimensions, and the otolith growth increment in the starved larvae was not deposited on a daily basis. After undergoing starvation, the larvae were unable to recover their normal growth either in otolith increment deposition or in body and otolith growth even though they were fed. Therefore, the application of ageing techniques based on counting otolith growth increments seems to be inaccurate for starved larvae.     

Assessment of the nutritional status of field-caught larval Pacific bluefin tuna by RNA/DNA ratio based on a starvation experiment of hatchery-reared fish

RNA/DNA ratio is a useful and reliable indicator of the nutritional status of fish larvae and juveniles. In order to assess the nutritional status of field-caught larval Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel), starvation experiments of hatchery-reared larvae were conducted and changes in the RNA/DNA ratio of fed and starved larvae were analyzed. Starvation experiments were conducted every 3 days after first feeding. The survival rate of Pacific bluefin tuna larvae ranged 10-50% after 1 day of starved conditions and growth retardation was observed immediately. These results suggest that Pacific bluefin tuna larvae have a very low tolerance to starvation. The RNA/DNA ratios of fed larvae were approximately 2.0-4.0. On the other hand, the value of starved larvae significantly decreased to 1.0-3.0. The nutritional status of 3 cohorts of field-caught tuna larvae collected in the northwestern Pacific Ocean was examined based on the value of the RNA/DNA ratio of the 1 day starved larvae. 4.35-25.77% of the cohorts were regarded as the “starving condition”, which was negatively correlated to the ambient prey densities. These findings suggest that the nutritional condition of larval Pacific bluefin tuna was influenced by the ambient prey density, and starvation itself and starvation-induced predation could greatly contribute to mortality in the larval period of Pacific bluefin tuna.     

Experimental influence of pH on the early life-stages of sea urchins I: different rates of introduction give rise to different responses

Many early life-stage response studies to ocean acidification utilize gametes/offspring obtained from ambient-sourced parents, which are then directly introduced to experimentally altered seawater pH. This approach may produce a stress response potentially impacting development and survival. Hence, this study determined whether this approach is suitable by subjecting embryos/larvae to different rates of introduction to lowered seawater pH to assess larval success under acute and staggered experimental pH scenarios. Embryos and 4-armed larvae of the sea urchin Psammechinus miliaris were introduced to pH conditions, widely used in ocean acidification studies, from ambient conditions utilizing 380, 470, 560, 700 and 840?ppm CO₂ changed at incremental steps at two rates: fast (every 3rd hour) or slow (every 48th hour). Direct transfers from ambient to low seawater pH gave rise to dramatic negative impacts (smaller size and low survival), but slower rates of introductions gave rise to lesser negative responses (low survival). There was no treatment effect on settled juveniles. Fast introductions utilized in many studies are likely not ideal approaches when assessing pre-settlement larval developmental responses. Therefore, careful consideration of the pattern of response is needed when studies report the responses of offspring, derived from ambient conditions, introduced directly to forecasted ocean acidification conditions.     

Evaluating the nutritional condition of individual whitefish (Coregonus spp.) larvae by the RNA/DNA ratio

Larvae of Coregonus spp. were reared in the laboratory under different temperature (4, 6 and 8°C) and feeding conditions ( ad libitum , limited, with starvation intervals). Their RNA/DNA ratios were determined with a highly sensitive fluorescence technique. After resorption of their yolk reserves (about 2 weeks after hatching), well fed larvae (RNA/DNA >2.5) could be significantly distinguished from larvae reared under limited food supply (RNA/DNA < 2.5), both at the 6 and 8°C levels. At 4°C, no differences due to the feeding regime were found. During a second series of experiments, larvae were affected by an intestinal disease, which was caused by the ingestion of unsuitable copepod plankton. This disease provoked high mortality, decreased growth and RNA/DNA ratios which were almost as low as in temporarily starved larvae from the first series. Coregonid larvae sampled in Lake Constance during spring 1990 showed RNA/DNA ratios which were unexpectedly low when judged on the basis of mean body length and average ambient temperature. It was obvious from macroscopic observations that some of these wild larvae were severely attacked by the intestinal disease. The low RNA/DNA ratios in field samples are, therefore, interpreted as a sublethal result of this disease.     

温度对花斑副沙鳅仔鱼发育、摄食及不可逆点的影响

2011年4～5月,研究了温度对花斑副沙鳅(Parabotia fasciata)早期仔鱼的发育、初次摄食时间、群体摄食率及不可逆点(PNR)的影响。结果发现,花斑副沙鳅仔鱼出膜后2 d已开始摄食;初次群体摄食率最高值出现在卵黄囊耗尽后的1～2 d,花斑副沙鳅仔鱼的摄食强度随温度升高而增大。仔鱼抵达PNR的时间随温度升高而缩短:22℃时,仔鱼的PNR出现在出膜后的8.0～9.0 d;26℃时,PNR为7.0 d;28℃时,PNR为5.5 d;30℃时,PNR为5.0 d。最大初次摄食率至PNR之间为不可逆饥饿期,22℃时为3.5 d,30℃时仅为1.5 d,说明温度越高,仔鱼耐受饥饿的能力越差,越不利于仔鱼的存活。研究认为,温度对花斑副沙鳅仔鱼发育、摄食及不可逆点的影响较大;在苗种培育过程中,应根据温度确定投饵时间,适时投饵对仔鱼的成活显得尤其重要。