方格星虫繁殖生物学研究

通过观察体腔生殖细胞发育的季节变化，研究广西海区方格星虫繁殖周期。研究样本采自北海市大冠沙至白虎头海区3个站点，1999年1月至2000年2月每月定期采集。观察结果表明，方格星虫雌雄异体；体腔内的生殖细胞最初成团附着在具有运动能力的滋养细胞上，卵原细胞经过细胞增殖期（1-2μm）、细胞质生长期（3-30μm）、滤泡膜形成期（30-70μm）、胶质膜形成期（70-160μm）和成熟期(≥160μm）5个发育时期；精原细胞的发育经过细胞增殖期、细胞生长期、细胞分化期和成熟期4个发育时期。方格星虫体腔全年都有生殖细胞存在，卵细胞分期成熟，分批产卵。广西海区方格星虫繁殖期于每年4-9月，繁殖盛期为5-8月。     

盐度对不同规格可口革囊星虫耗氧率和排氨率的影响

水温18.5~20℃下,采取实验生态学方法,测定了10、15、20、25、30和35盐度下3种规格[干质量(0.220±0.000)、(0.489±0.002)g和(0.899±0.002)g]可口革囊星虫的耗氧率和排氨率。试验结果表明,盐度、规格均显著影响可口革囊星虫的耗氧率和排氨率(P0.05)。在盐度10~25时,3种规格可口革囊星虫的耗氧率和排氨率均随盐度增加而增大,盐度25时达到峰值后,随着盐度增大而减小。耗氧率和排氨率与盐度呈一元二次函数关系:y=ax~2+bx+c。在相同盐度下,可口革囊星虫的耗氧率和排氨率随体质量的增大而显著减小(P0.05),且耗氧率和排氨率与干质量符合y=amb模型。盐度10~35时,可口革囊星虫的氧氮比值为5.947~25.246,表明可口革囊星虫主要由蛋白质和脂肪提供能量。     

可口革囊星虫对温度、盐度的耐受性试验

可口革囊星虫(Phascoloma esculenta)属星虫动物门、革囊星虫纲、革囊星虫目、革囊星虫科、革囊星虫属,是我国的特有种,主要分布于广东、广西、海南岛、福建、浙江沿海,有些地方的潮间带栖息密度较大[1-2]。可口革囊星虫肉味鲜美、营养丰富,加工成品“沙虫干”是著名的海产珍品之一,深受国内外消费者欢迎;做成“土蒜冻”制品具有滋阴降火、补肾益智、泻火的功效[3-4]。目前,有关可口革囊星虫的生物学基础资料较少[5-7],笔者探讨温度和盐度变化对可口革囊星虫的影响,从而为可口革囊星虫的人工养殖和人工育苗提供理论依据。1材料与方法1.1材…     

可口革囊星虫规模化生态繁育试验

为缓解可口革囊星虫苗种紧缺的现状,探索其批量苗种繁育技术,于2011~2013年在浙江省温岭市利用潮间带滩涂模拟自然生态建立生态繁育区,面积由110亩(15亩=1 hm~2,下同)扩大到210亩,开展了可口革囊星虫生态繁育试验。2013年,在繁育区选取20亩作为放养区,放养规格462尾/kg的亲体2 625 kg,折合放养密度6.064万尾/亩,2014年6月,对210亩生态繁育区自然繁育的星虫苗种进行测产,测得星虫苗种数量为3.375亿尾,规格为2 753尾/kg,3年累计培育可口革囊星虫苗种7.174亿尾。数据显示,可口革囊星虫的生态繁育试验收效甚佳,如能进一步扩大生态繁殖规模,将在一定程度上缓解可口革囊星虫苗种紧缺的现状。     

光裸方格星虫繁殖习性的初步观察

2011年6、7、8月每月10日,挑选♀∶♂为5∶1的性成熟光裸方格星虫(Sipunculus nudus)亲体,在水泥池中暂养20 d,观察其产卵习性、产卵量及其体腔中生殖细胞的变化。结果表明,光裸方格星虫雌雄异体、体外受精;卵母细胞分批成熟,分批产卵;产卵时间不固定,昼夜皆可产卵,但夜间产卵次数和产卵量均高于白天;大量产卵时集群游动;6、7、8月的雌光裸方格星虫平均产卵分别为:3.20×104、4.43×104、3.46×104cell,平均受精率达98.3%;6、7、8月日累计产卵量y与时间x的变化方程分别为:y=0.430 5x3-25.971x2+503.39x-582.56(R2=0.982 0)、y=0.583 3x3-33.942x2+659.04x-712.43(R2=0.984 7)、y=-0.31x3+1.4676x2+245.11x-278.22(R2=0.994 3)。6、7、8月成熟卵细胞相对百分含量分别降低4.8%、9.0%、26.0%(P<0.01);6、7月的光裸方格星虫产后具有较好的再繁殖能力。     

重金属镉和锌对可口革囊星虫的毒性试验

研究了镉、锌离子对可口革囊星虫的单一和联合毒性。结果显示:镉离子对可口革囊星虫的48 h和96 h半致死浓度分别为501 mg/L和175 mg/L,锌离子对可口革囊星虫的48 h和96 h半致死浓度分别为82 mg/L和61 mg/L,镉和锌对可口革囊星虫的安全浓度分别为17.5 mg/L和6.1 mg/L;镉和锌对可口革囊星虫的联合毒性在48 h和96 h时AI<0,相互拮抗作用明显。     

可口革囊星虫高密度养殖技术

正可口革囊星虫俗称泥丁,是生长在潮间带高潮区、泥沙底质地带的经济类星虫,主要分布于我国南海沿岸海滩,红树林底下的滩涂是它的最佳栖息地。笔者对可口革囊星虫育苗及养成的探索始于2012年,现将多年累积的经验与大家分享。一、可口革囊星虫养殖场地的选址可口革囊星虫以海底的有机碎屑、底栖硅藻     

星虫动物线粒体基因组全序列的基因排列和特征比较

综合利用通用引物PCR和长PCR扩增,获得可口革囊星虫的线粒体DNA,结合鸟枪法和引物步移法获得可口革囊星虫的线粒体基因组。比较可口革囊星虫与方格星虫线粒体基因组,所有的37个基因均在同一条链上编码。2个星虫动物线粒体基因组蛋白质编码基因的起始、终止密码子及氨基酸长度存在差异。2个星虫动物的线粒体基因组,存在6个保守的基因区块:(1)nad6-cob-P-S2-C-M,(2)I-K-nad3-F,(3)nad4L-nad4-L2,(4)nad1-W-atp6,(5)nad5-S1及(6)nad2-cox1。去除转运RNA而仅比较蛋白质编码基因和核糖体RNA,则2个星虫动物线粒体基因组主编码基因的基因排列完全一致,均为:cox1-cox2-atp8-cox3-nad6-cob-srRNA-lrRNA-nad3-nad4L-nad4-nad1-atp6-nad5-nad2。星虫动物、环节动物和螠虫动物线粒体基因组共有5个保守区块:(1)cox1-cox2-atp8,(2)nad4L-nad4,(3)srRNA-lrRNA,(4)cox3-nad6-cob,(5)atp6-nad5。在15个主编码基因中,nad5基因和nad4基因变异位点数最多,因此可作为备选的分子标记,用于分析星虫动物不同物种和群体之间的生物多样性。     

可口革囊星虫对海水盐度耐受力的初步研究

在室内条件下,可口革囊星虫对海水盐度耐受力的实验表明,不同个体星虫对盐度的耐受力没有显著差异,三组平均成活率均大于90%.可口革囊星虫存活的适宜海水盐度范围为10～35.本实验结果发现该种对低盐海水适应性很强,验证了该种的自然分布和生态习性.测定113条星虫体重与体壁干湿重关系,其直线回归分别为y=0.2819x-0.0142(R2=0.9529)、y=0.1886x 0.0028(R2=0.9891).     

可口革囊星虫体壁形态和组织学结构

试验结果表明,可口革囊星虫吻长与躯干长之比介于1与2之间,体壁由外到内依次由角质层、上皮、环肌层、纵肌层和壁体腔膜组成。角质层是由上皮细胞分泌而成的非细胞结构,外突形成乳突且在吻顶端特化为钩,乳突顶端含有棕色颗粒物质;上皮薄,为单层柱状上皮;肌肉发达,外环内纵,纵肌成束状分布;体壁的最内为壁体腔膜,由单层上皮细胞组成,朝体腔侧有具纤毛细胞的存在。AB-PAS染色显示体壁仅有少量混合黏液细胞存在。     

福建省东山岛西埔湾港养斑鰶的繁殖生物学

本文研究了福建省东山岛西埔湾港养斑鰶性腺周期发育、成熟系数周年变化、个体生殖力以及雌雄同体。港养斑鰶能在半封闭式湾内发育成熟而且自然产卵。其卵细胞发育分为6个时相,精巢发育分为6期。卵膜由透明带、外放射带和内放射带组成,内有孔道结构。第4时相中期卵母细胞的内放射带有7～8层纤维状片层。产卵后鞘膜细胞和颗粒细胞分裂增殖变成游走的吞噬细胞。西埔湾港养斑鰶Ⅰ龄鱼初次性成熟,生殖季节从12月下旬至翌年3月下旬,生殖盛期为2月,雌鱼成熟系数高达17.88％。个体绝对生殖力为7575～72621粒,平均 35310粒,系属于分批产卵类型,第1批生殖力大于第2批。     

Reproductive parameters of the chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> estimated from human chorionic gonadotropin-induced final oocyte maturation and ovulation in captivity

ABSTRACT:   Final oocyte maturation and ovulation of captive chub mackerel Scomber japonicus with fully yolk-accumulated oocytes were induced by a single injection of human chorionic gonadotropin. Reproductive parameters, including spawning frequency and batch fecundity, which are required to estimate spawning biomass in pelagic fish by the daily egg production method, were analyzed. Germinal vesicle migration (GVM) occurred at 18–24 h post-injection, and the hydration and ovulation of oocytes were completed at 30 and 36 h post-injection, respectively. The results of the maturation process suggest that fish with GVM-stage ovaries captured in the daytime from the field are capable of spawning on the night following their capture. The oocytes used in the oocyte size-frequency distribution method for batch fecundity estimates should be at late GVM and more advanced stages. The results of sequential artificial insemination showed that the quality of ovulated eggs held in the ovarian lumen rapidly deteriorated as time progressed after ovulation. This indicates that the fertilization window for the ovulated eggs of chub mackerel lasts only a few hours, and spawning behavior should be performed within a few hours after ovulation in the wild population.     

Oogenesis and lipids in gonad and liver of daubed shanny (Leptoclinus maculatus) females from Svalbard waters

Oocyte and liver histomorphology of the daubed shanny (Leptoclinus maculatus) from Isfjorden and Kongsfjorden in Svalbard were investigated during three Arctic seasons: summer (July), autumn (October) and winter (April). Three oocyte developmental phases were observed: primary growth phase, secondary growth phase and maturation phase. We observed four different developmental stages: (1) perinucleolus stage with cortical alveolus formation, (2) lipid droplets formation, (3) vitellogenesis stage and (4) maturation. Late maturation stage of oocytes in the ovaries was from the autumn season. Females accumulated lipids in liver (up to 35.2?% dw) and deposited large amounts of lipids into gonads (up to 52.2?% dw) during maturation. Lipid classes in female gonads changed seasonally, with relative increase in cholesterol during summer and depletion of storage lipids (triacylglycerols and wax esters/cholesterol esters) during the winter. Lipid composition in liver changed during oocyte development and spawning, as neutral lipids were transferred to developing oocytes during summer to autumn. During winter, storage lipids were depleted during starvation. Based on the increase in gonadosomatic index (GSI) with length and observed maturation stages, females seem to mature at a length of about 125-130?mm. The GSI and hepatosomatic index of large females sampled in autumn (September-October) were significantly higher than for females in late winter (April) and spring (May). These results indicate that spawning takes place during winter in Isfjorden and that energy reserves stored in the liver are utilized by females during gonadal development and reproduction.     

Stages of gonad development during the reproductive cycle of the blackfin snook, Centropomus medius Günther

The reproductive biology of the Blackfin snook Centropomus medius was studied to expand our current knowledge on its aquaculture potential. From April 2001 to November 2002, 301 females and 193 males were captured. A 1:1.6 (male:female) ratio was found. Gonadosomatic (GSI) and hepatosomatic (HSI) indices suggested that the spawning period (SP) extended from September to November, with peak spawning activity in September (GSI=0.98% and HSI=0.7%). The period of pre‐gametogenesis extended from January to June. During the post‐SP in December, 90% of oocytes were pre‐vitellogenic and 10% were atretic. Histological analysis of gonads (haematoxylin–eosin, Sudan black B, Alcian blue PAS, Cleveland Wolfe) showed five stages of oocyte development, each occurring at a characteristic range of oocyte diameters (μm): (I) pre‐vitellogenic (25–55); (II) cortical alveoli (56–250); (III) vitellogenic (251–340); (IV) maturation (341–645) and (V) atretic (21–100). Five reproductive stages were identified for testis morphology: (I) regressed; (II) early maturation; (III) mid‐maturation; (IV) full maturation and (V) regression. The type of ovarian development in C. medius was identified as a group synchronous.     

Maturation and reproductive cycle of female armorhead <Emphasis Type="Italic">Pseudopentaceros wheeleri</Emphasis> from the southern Emperor—northern Hawaiian Ridge Seamounts

ABSTRACT:   The stage of ovarian development and annual reproductive cycle of female armorhead Pseudopentaceros wheeleri collected from seamounts of the southern Emperor–northern Hawaiian Ridge in 1993 and 1995–1996 was investigated through use of the gonadal somatic index ( GSI ) and ovarian histology. The stages of oocyte development were classified into eight developmental stages (perinucleolus, cortical alveolus, oil droplet, early yolk globule, late yolk globule, migratory nucleolus, prematuration, and maturation) based on histological characteristics. Oocytes of different size and developmental stage were present within ovaries during the spawning season. Oocyte development appears to be asynchronous and armorhead apparently spawn several times during the spawning season. The stage of the most developed oocyte was used to classify the stage of ovarian maturation. The GSI increased gradually with ovarian development. Oocytes in the perinucleolus, cortical alveolus, and oil droplet stages were present in ovaries throughout the year. Early and late yolk globule stages appeared during August–September indicating that yolk formation begins during late summer. The migratory nucleolus through maturation stages were detected during November–January. Post ovulatory follicles were present during November–March, and GSI values rapidly decreased in February. Results indicate that spawning occurs during November–February and peaks during December–January.     

Histological examination of final oocyte maturation and atresia in wild and domesticated Penaeus monodon (Fabricius) broodstock

Oocyte maturation and gonadosomatic index (GSI) of eyestalk ablated Penaeus monodon females collected from the wild and from two first‐generation domesticated lines were assessed. Frequency and diameter of the different oocytes, and the intensity of oocyte atresia, were compared among groups through histological assessments of the sections of the middle ovarian lobe. Digitized images from ovary sections were used to record the frequency and diameter of different oocyte types. Spawning performance of the three groups were expressed in terms of the percentage of females that spawned at least once (productive females), time from eyestalk ablation to first spawning (latency period) and the number of spawnings per female stocked. Final ovarian maturation was attained in all groups, as indicated by the presence of mature oocytes with cortical rods (cortical oocytes), dark‐green ovarian colour and high GSI values (5.83–6.86%). However, domesticated groups presented significant larger immature oocyte types (previtellogenic and yolky oocytes) and smaller cortical oocytes compared with wild females, indicating a reduced vitellogenic activity during final oocyte maturation. Additionally, the frequency of atresia was comparatively higher for both domesticated groups, which could be related to their inferior spawning performance. The implications of these results on the reproductive potential and development of domesticated P. monodon are discussed.     

Induced spawning of red drum,Sciaenops ocellatus: use of multivariate and univariate analysis methods in the search for side effects of LH-RHa treatments and ovarian development state upon spawn quality

The present work examines the usefulness of the combination of multivariate analyses and further one-way analysis of variance in the search for links between causes and effects in heterologous stimulation of spawning in fish. Stimulations with both 20 and 50 μg.kg^–1 doses of [D Trp⁶] luteinizing hormone-releasing hormone analogue (LH-RHa) were effective in inducing and synchronizing spawning of red drum, Sciaenops ocellatus. Oocyte development monitoring using ovarian biopsy allowed treatment injections at different stages of maturation. The influence at stimulation time of female oocyte development state upon further development was shown. An interesting potential of second stimulation, leading to high quality spawns, was established and factors of success were identified. The best conditions for stimulation success were simultaneously late stage of oocyte development characterized by the beginning of oil droplet coalescence, high ovarian homogeneity quoted by the proportion of most developed oocytes in the biopsy sample, and low dose of hormone (20 μg.kg^–1) for both first and second stimulations.     

Interaction “nutrition-reproduction” in fish

In wild fish, gonad maturation and spawning induce modifications in feeding behaviour and dynamics of body stores which are possibly responsible for metabolic disorders. Conversely, food manipulations can modify reproduction of captive female broodstock. A restricted diet during early stages of the life cycle delays the first maturation age, a quantitatively restricted food supply during the stages of oocyte differentiation reduces the egg number, while a reduction of food levels during the last phases of oogenesis has only a small effect on egg size, composition and hatchability. During the spawning period, salmonid female broodstock seem to be more able to utilize carbohydrates than immature fish, they valorize the energy brought in by fats and have low quantitative requirements for proteins (28%). On the other hand, Red Sea bream broodstock have higher needs for proteins (45%).The incorporation into diet of liposoluble vitamins and essential fatty acids is necessary for normal reproduction, but (n-6) fatty acids seem to play a more important role than in juveniles. Although the effects and physiological role of carotenoids are not yet well known in salmonids, they are necessary for normal development in Red Sea bream. Although little work has been done on the effects of mineral nutrition, special attention must be paid to trace elements, manganese in particular.The major part of this review concerns female broodstock, since very little interest has been devoted to males.     

中华绒螯蟹卵黄发生期卵母细胞和卵泡细胞超微结构观

通过透射电镜技术观察了中华绒螯蟹第二次卵巢发育过程中卵巢的超微结构变化。结果表明：（1）中华绒螯蟹第二次卵巢发育过程中卵黄发生期可分为初期和后期；（2）卵黄发生初期（雌蟹第一次排卵后的16 d内），卵黄生成以卵母细胞内源性合成为主，此时卵母细胞胞质中存在大量内质网囊泡、高尔基体和线粒体，这些细胞器参与胞内卵黄物质的合成。内源性合成后期，卵母细胞膜形态多样，呈现触手状、波浪状和断裂状，为外源合成期做准备。此期卵泡细胞还未向卵母细胞靠近，两类细胞间存在着由淋巴细胞吐出的絮状物；（3）卵黄发生后期，首先为卵泡细胞与卵母细胞的结合阶段（排卵后16~21 d），此后，卵泡细胞胞质中含有大量内质网囊泡、卵黄颗粒和脂滴，卵母细胞与卵泡细胞膜变为链珠状便于物质交换，卵母细胞的卵黄合成能力减少，转由卵泡细胞进行外源性物质吸收和卵黄物质合成（21~36 d）；（4）卵黄发生结束后，双层卵膜形成，卵黄体和脂肪滴均匀分布在卵母细胞胞质中。