环境条件对中国对虾交尾影响的试验观察↑（*）

本文报告了在室内条件下中国对虾雄性生殖系统不同部位精子形态、激活反应及外界环境对对虾交尾影响等的试验观察。雄性生殖系统被分为３部分：精巢（Ｔ）、输精管和精荚（ＴＡ）;输精管又分为前、中、后３段,其中的前（ＡＶＤ）、中（ＭＶＤ）、后（ＰＶＤ）３部分以及精巢和精荚被分别匀浆取样镜检,观察精子形态。结果表明：中国对虾雄性精巢、输精管、精荚及雌虾纳精囊内精子的形态、对卵水的激活反应能力有所差异,是一逐渐成熟过程;雄虾具有多次交尾能力,不同环境条件对中国对虾的交尾影响明显;当雌虾比例较高时交尾率也高;不同对虾密度对交尾影响不明显     

扇贝精子及卵子的受精生物学特性

采用扫描电子显微镜和普通光学显微镜观察方法,结合全光谱扫描分析方法,对栉孔扇贝和虾夷扇贝的精子及卵子的受精生物学特性进行了详细的观察,包括精卵产出后形态学变化、卵水的特性、精子顶体反应、卵子皮层反应以及精子分级分离组分对卵子的作用5个方面.结果表明,这两种扇贝精卵生物学特性没有本质的区别,扇贝精子排入海水中10min以内,外形没有太大的变化,但30min以后约有1/4的精子出现头部膨大变成圆球形的现象,受精能力明显下降;扇贝卵子产入海水中1h以内受精能力没有变化,但2h以后,约1/3的卵子出现卵裂现象,未见极体的排放,且卵裂多停止在2细胞期,少数达到4细胞期,基本属于均等卵裂,并失去受精能力;滴入卵水中的精子极易发生自溶解体,未解体的精子发生凝集,10min之后卵水中基本检测不到完整的精子,卵水和钙离子载体A23187均能诱导精子发生顶体反应;精子分级分离组分分别为精浆、精子头部和精子尾部,这三部分除了少数精子头部能够附着在卵子表面外,其他都不具备激活卵子的作用.对扇贝精子和卵子的生物学特性的研究将为解释不同种扇贝的精卵相互识别并受精的现象提供基础资料.     

人工控制自然交尾条件下中国对虾父本的微卫星识别

人工控制自然交尾条件下,在保持中国对虾雌、雄5∶1的交配比率下,获得了两尾交尾并产生子代的雌虾,同时有4个疑似父本需要识别.利用3个微卫星引物及其组成的三重PCR技术对4个疑似父本进行了鉴别,准确的找到了相应的与雌虾交尾的雄虾.这为建立半同胞家系提供了一种新的技术和思路.     

利用塑料大棚延长对虾交尾期的研究

为提高对虾交尾率，１９９３，１９９４年进行了塑料大棚延长对虾交尾期试验，通过大棚控制交尾水温，采取合理安排雌雄比，调节水质，投喂沙蚕，防治疾病等综合技术措施，２年共出棚交尾虾１２３５２尾，未交尾虾５９２２尾，雄虾５２５２尾，平均交尾率６７．５９％，雌雄虾成活率分别为８７．５４％，６３．００％，共获产值６２６１００元，纯利４４２６００元，平均每尾入棚雌虾净协效益２１．２３元，专家鉴定认为，本研究达国     

克氏螯虾繁殖性状初探

一、材料与方法 　　1.材料来源 　　本次试验所用克氏螯虾，全部从洪湖蓝田生态养殖二场二万亩大湖圈养捕捉而来，且捕捉时间不同。当时捕捉收集的虾当时解剖，共解剖虾 187只，其中雌虾 103只，雄虾 84只。未被解剖的虾放入暂养池中，观察其繁殖行为和抱卵期行为变化情况。 　　2.方法 　　对试验虾分雌雄两组，先称体重，然后对头部去壳观察其性腺，并对性腺称重。对雌虾的卵先用显微镜观察其形状，再观察其所处时期，然后用 Boun’ s液固定后人工计数，确保准确；对雄虾性腺称重，解剖后用显微镜观察精子形状和活力。 　　二、结果 …     

影响中国对虾(Penaeus Orientalis)室内交尾的主要因素

<正> 对虾室内交尾率低,限制了对虾大规模越冬生产。研究实践表明: 影响对虾室内交尾的主要因素如下: 1.生物节律。海虾和养殖虾的交尾活动都具有明显的季节性和日节律,在自然海区对虾交尾从10月中旬开始,到11月上旬结束。交尾期大约两周左右。交尾高峰一般在10月下旬。对虾的蜕皮与交尾都集中在大潮汛期,阴历九月初的大潮汛中有82％的雌虾交尾,阴历九月     

日本蟳精子超低温冷冻保存技术的研究

以精子存活率作为评价指标,采用两步降温法研究了稀释剂、抗冻保护剂和预冷时间对日本蟳精子超低温冷冻保存效果的影响。结果表明:以采用稀释剂II、15%二甲基亚砜(DMSO)作为抗冻保护剂的保存效果最佳。在液氮中保存24 h后,精子存活率可达83.76%,保存一年后可达73.81%。精液的第一次预冷时间以25 min为宜。     

影响中国对虾交尾率与亲虾越冬成活率的主要因素

一、影响中国对虾交尾的主要因素获取足量的交尾且体质健壮的雌虾是亲虾人工越冬生产的根本,而对虾交尾活动又受季节、水温、盐度和光照等若干因素的影响。因此,掌握对虾交尾活动的生态规律,是提高亲虾人工越冬生产技术水平的重要基础。     

电刺激日本沼虾精荚排放及人工授精的研究↑（*）

应用低压交流电刺激日本沼虾精英排放，进行精英移植，取得了人工授精的成功，在电压为４－６Ｖ范围内，刺激虾体精英排放的效果最佳，而对虾体无伤害；间隔４８小时的电刺激，可再次获得精英，且精英中精子的形态结构正常。精英移植人工授精实验结果表明，经精英移植的１６只已完成生殖蜕皮的雌虾中，１０只顺利产卵，受精率高达７１％。     

红螯螯虾的室内产卵和卵发育

１９９１～１９９２年对澳大利亚北方的热带大型虾红螯螯虾ｃｈｅｒａｘｑｕａｄｒｉｎａｒｉｎａｔｕｓ在室内进行了孵化实验。６个玻璃钢水槽放养４０～１９０ｇ的性成熟红螯螯虾１０只／ｍ２，雄雌比率１：２，水温保持在２８±１℃。１９９１年采用自然光照，１９９２年控制光照。采用自然光照，每日光照不是１２小时．月产卵率为１５％；每日光照有１２小时以上，月产卵率在３５％以上。由于产卵高峰期在５～７月，故春季放养为时过晚。翌年，将光照由１２月的１０小时光照１４小时黑暗调节至２月的１４小时光照１０小时黑暗，且一直持续至７月。红螯螯虾产卵率随光照期的延长而提高，且产卵高峰期提前在３～５月，月产卵率高达５０％。５月份以后，即使水温和光照期一直不变，产卵率将直线下降。所产卵数与雌虾个体规格呈正相关。４０～１９０ｇ的雌虾平均产仔虾７．３只／克体重，卵色用于预测卵的发育期。     

Egg Water Induced Reaction and Biostain Assay of Sperm from Marine Shrimp Penaeus vannamei: Dietary Effects on Sperm Quality

Reproductive performance was evaluated for sexually mature male Penueus vannamei fed one of four diets: 1) commercial 40% protein shrimp growout diet; 2) 100% squid; 3) 50% squid, 25% bloodworms and 25% Artemia; and 4) starvation. Spermatophores were ejaculated manually from tagged males at stocking and every 12 d thereafter. For each half of the compound spermatophore, weight and sperm count were determined. Four methods were used to assay the sperm quality: 1) gross morphology (GM); 2) trypan blue stain (TB); 3) acridine orange stain (AO); and 4) egg water induced reaction (EW). The reaction induced when P. vannamei sperm are exposed to conspecific egg water is grossly similar to that observed in vivo at the surface of the freshly spawned eggs from naturally mated females. Little correlation was found between spermatophore weight and sperm count. The best correlation was found between GM and AO (r²= 0.859, P < 0.01). Many morphologically abnormal sperm did not stain with TB. For most of the spermatophores evaluated; over 80% of the sperm were morphologically normal, were unstained with TB, and were immediately fluorescing light green when stained with AO. When exposed to egg water, sperm from these apparently good quality spermatophores yielded mixed results. The analysis of dietary effects on spermatophore quality were equivocal. Although a significant decline in body weight, spermatophore weight, and sperm count were observed in the starvation treatment as compared to maturation and squid diets, no significant differences were observed in MY of the sperm quality assays.     

Acrosome reaction in the sperm of the black tiger shrimp Penaeus monodon (Decapoda, Penaeidae)

Owing to the problem of male infertility in the domesticated shrimp Penaeus monodon, this study was conducted to reveal the morphological events of an acrosome reaction (AR) of sperm of this highly valuable species. The AR observed in an in vitro incubation of sperm with egg water (EW) and that during actual spawning was compared. Under transmission and scanning electron microscopy, sperm taken from the female thelycum was composed of a posterior main body, a central cap and an anterior single spike. Upon contact with EW, the sperm underwent two phases of AR: acrosomal exocytosis and spherical mass formation. The former was composed of a degeneration of the spike, swelling of the cap region and rupture of the acrosomal pouch. The latter began with polymerization of materials within the subacrosomal region and ended with re‐configuration of the subacrosomal region into an electron‐dense spherical mass. The AR of the sperm observed during spawning revealed similar morphological events, with degeneration of the spike upon contact with the vitelline envelope and formation of the spherical mass while penetrating into jelly material produced by protruding cortical rods. The results suggest the presence of AR inducers derived from the vitelline envelope and cortical rods of the egg. This study forms the basis for an evaluation of infertility regarding to AR in the domesticated P. monodon male.     

Biochemical characterization and physiological role of cortical rods in black tiger shrimp, Penaeus monodon

Cortical rods (CRs), precursors of egg jelly investment in many penaeoid shrimp, are composed of different proportions of proteins and carbohydrates, the physiological role of which still requires extensive investigation. In this study, we demonstrated the biochemical properties of the CRs and their role in the induction of the acrosome reaction (AR). Profiles of the isolated CRs revealed a number of major protein bands ranging from 35 to 230 kDa. These CR proteins were extensively glycosylated and sulfated. Lectin-based carbohydrate analysis further revealed the highest reactivity of concanavalin A (Con A) among other lectins used. In addition, the selective interference of Con A binding with mannose but not glucose indicated that CR glycoproteins were of high-mannose type. Using immunoblotting with anti-CR antibody, we further demonstrated that part of egg water (EW, a natural AR inducer) was derived from miscible components of the CRs. Physiological tests of water-soluble CR (wsCR) revealed its high AR inducing competency comparable to that of EW, which was far superior to that of acid-urea treated CR (auCR). Furthermore, the wsCR-induced AR was selectively inhibited by Con A, suggesting the significance of the exposing mannose residues in regulating P. monodon sperm AR response.     

史氏鲟精子入卵过程的扫描电镜观察

史氏鲟(Acipenserschrencki)隶属鲟形目(Acipenseriformes)、鲟科(Acipenseridae)、鲟属(Acipenser),是黑龙江特产和重要的经济鱼类之一,为淡水定居种类。鲟鱼是古老的生物类型,世界上约有28种,仅分布于北半球。由于自然环境的改变和人类活动的影响,所有鲟鱼均处于不同程度的濒危状态,有的已近灭绝[1,2]。我国有8种鲟鱼,其中黑龙江2种,即史氏鲟和达氏鳇。近年来,随着人工养殖技术的提高和完善,史氏鲟成为国内最主要的鲟鱼养殖种类。有关史氏鲟人工养殖、基因组DNA含量、胚胎发育等国内有较多研究,而对受精生物学方面研究尚未见报道。特殊…     

水温和盐度对南美白对虾幼虾能量收支的影响

用室内生态实验法测定了南美白对虾幼虾在不同盐度(5,15或25)和水温(20～23℃,25℃,30℃或33℃)下摄食水丝蚓的能量收支方程。在盐度5～25时,随盐度降低,对虾的能量转换效率升高,同化率降低。在盐度为5时,K1最高(29.02%),同化率最低(77.89%)。对虾具补偿机制来补偿调节渗透压的能耗。盐度对同化率的影响,主要是通过影响呼吸代谢实现的。对虾的特定生长率(y)(%·d-1)随水温(X)(℃)上升而增加,其关系式为:y=0.0168LnX 0.0831(R2=0.9324)。对虾在较高温度下生长快,主要是增加绝对摄食量,提高吸收效率,减少粪便排泄量。在相同温度下,变温时对虾的摄食量高于恒温时。     

MATURATION,REPRODUCTION, AND GROWTH OF Penaeus vannamei AND P. stylirostris FED NATURAL DIETS

A variety of natural foods have been reported to induce ovarian maturation of penaeid shrimp. In this study, conducted from 19 June to 11 September 1980, four single-food diets (clams, shrimp, squid, and worms) and one composite diet consisting of all four foods were fed to Penaeus vannamei in 3.7-m diameter indoor circular tanks. The shrimp, averaging 28 g, were stocked at a rate of 6.9 m^-2 with a 1:1 male to female ratio. Unilateral eyestalk ablation was performed on half of the females in each tank. Tanks received one turnover of water volume daily and the water temperature and salinity averaged 27.0°C and 28.6 ppt, respectively. Daily photoperiod was maintained at 13 hours of light. Ovarian maturation, spawning, molting rate, and survival were monitored daily and growth and gonad size were measured at termination. The composite diet was the best overall diet, while squid was the best single-food diet, followed by shrimp, worms, and clams. No maturation of unablated females was observed with the exception of one female that developed to stage 3. Although the size of the shrimp was considered marginal for spawning, 5 mated females were observed, but only one (squid treatment) produced viable nauplii. After termination of this study, all shrimp were fed the composite diet for 41 days. During this period, ovarian maturation and spawning occurred in the previously unproductive treatment receiving only clams, and growth rate of shrimp in all previously single-food treatments increased 4 to 7 fold. The composite diet was also fed to two tanks of P. stylirostris (mean initial weight = 45.4 g) for a period of 118 days. During that time, 50 spawns were obtained with up to 3 spawns per female. A total of 4.3 × 10⁵ P. stylirostris nauplii were produced.     

THE ARTIFICIAL PROPAGATION AND CULTURE OF BEAR SHRIMP,Penaeus semisulcatus de HAAN,IN HONG KONG

Penaeus semisulcatus (bear shrimp) was used for biological and culture studies. A peak of reproductive periodicity was shown in the months of May, June and July. Increase in body weight and in gonad weight coincided with the increase in sea water temperature. This suggested that sexual maturation might be a direct response to increased sea water temperature. Fecundity of mature female shrimp was estimated as 415,000 to 479,000 ova. In most cases, half-spent spawnings led to the production of poor eggs, characterized by irregular cytoplasmic formation and final autolysis. The embryonic and larval development of P. semisulcatus proceeded satisfactorily in a slightly alkaline medium (pH range from 7.5–8.5), and in water of salinity ranging from 28 to 35 ppt for egg and nauplius stages, and thereafter 25–35 ppt for zoea and mysis stages. The shrimps exhibited faster increase in body length than body weight in early growth. Later, after the shrimps had reached a body length of about 7.0 cm, the growth rate of body weight increased more markedly than body length. The rate of daily increase in weight was 1.01% in 7.0 cm shrimp. The feed efficiency of the formulated shrimp pellets was found to be 31.4%. During the nutritional study of P. semisulcatus, it was found that the combination of high dietary protein (about 40%) and low dietary lipid produced best growth and survival of bear shrimp. High increase of biomass of the shrimp fed with clam meat and high quality fish meal demonstrated the favorable response of shrimp to these diets. The use of cages for culturing bear shrimp was found to be practicable, but not efficient.     

Artificial fertilization in turbot, Scophthalmus maximus (L.): different methods and determination of the optimal sperm–egg ratio

With the aim of improving artificial fertilization (AF) in turbot, Scophthalmus maximus (L.), a series of fertilization experiments was carried out under dry conditions and different wet conditions (eggs/sea water: 2V/V and V/V). Another series of fertilization experiments was carried out with different quantities of sperm pool to determine the optimal ratio of spermatozoa to eggs for each AF method. Sperm pool from two males and eggs from spawns with a viability rate of > 70% were used. The sperm pool’s density (0.4–5.18 × 10⁹ sperm mL^–1) and motility (1–5) had been assessed previously. Significantly different fertilization rates were found when comparing 2V/V and V/V wet conditions. Significantly higher fertilization rates were found in dry fertilization when the sperm–egg ratio was > 9000 spermatozoa per egg and, under wet condition V/V, at 3000–4000 spermatozoa per egg.     

Performance of Larval and Postlarval Pacific White Shrimp,Litopenaeus vannamei (Boone), Fed Two Commercial Liquid Diet Supplements

Abstract

Two commercial liquid diet supplements were evaluated as a partial replacement for live foods fed to larval and postlarval Pacific white shrimp, Litopenaeus vannamei. Larvae were stocked in 20-L buckets at a density of 113 shrimp/L and fed one of four diets (5 replicate buckets per diet): (1) a control diet consisting of the diatom, Chaetoceros neogracile, and live Artemia; (2) the control diet plus Treflan (a fungicide); (3) LiquaLife liquid larval diet supplements with reduced concentrations of C. neogracile and Artemia; and (4) Epifeed liquid larval diet supplements with reduced concentrations of Artemia. Diets were evaluated by comparing shrimp rostro-caudal length and survival to PL-8, survival to osmotic and pH stress, as well as shrimp gut fullness, gut lipid content, and fouling. Rostro-caudal length and survival to PL-8 were not significantly different among the four treatments. However, mean survival to PL-8 was 12-34% greater in the Epifeed treatment than in the other three treatments. Shrimp fed Epifeed exhibited greater survival (P < 0.05) when exposed to 7 ppt water for two hours than shrimp in the control treatments. In contrast, there was no significant difference in survival (P > 0.05) among the four treatments when shrimp were exposed to apH of 3.35 for two hours. Although shrimp fed liquid diet supplements had more fouling (P < 0.05) than shrimp in either of the control treatments, commercial liquid diet supplements can be used in penaeid shrimp hatcheries to partially replace Artemia without compromising shrimp growth or survival.