Ontogenetic development in the morphology and behavior of loach （Misgurnus anguillicaudatus） during early life stages

Loach （Misgurnus anguillicaudatus） is a commercially important fish in China and an ideal aquaculture species. However, culturists experience high larval and juvenile mortality during mass production. To provide insight into ways to improve larviculture techniques, we describe the morphological characteristics and behavior of loach during the larval and early juvenile stages. Yolksae larvae ranged from 2.8 to 4.0 mm body length （BL） between days 0 to 4; preflexion larvae ranged from 3.6 to 5.5 mm BL between days 4 to 6; flexion larvae ranged from 4.8 to 8.1 mm BL between days 5 and 14; and post-flexion larvae ranged from 7. l to 15.7 mm BL between days 11 to 27; the minimum length and age of juveniles was 14.1 mm BL and 23 d, respectively. Loach are demersal from hatch through to the early juvenile stages. A suite of morphological characteristics （e.g., external gill filament and ventral mouth opening） and behavioral traits have developed to adapt to demersal living. We observed positive allometric growth in eye diameter, head length, head height, and pectoral fin length during the early larval stages, reflecting the priorities in the development of the organs essential for survival. Our results provide a basis for developing techniques to improve the survival of larval and juvenile loach during mass production.     

Isolation and cultivation of the vegetative cells ofPorphyra haitanensis

This research deals mainly with the use of the vegetative cells of a marine red algaPorphyra haitanensis (laver) as seeds and their culture into young thalli for further cultivation in the sea. The experimental process was as follows. Enzymatically isolated vegetative cells were attached to rope substrates and cultured in the laboratory for a month until they became about 0.2–0.5 cm long sporelings and were then attached (7–15 sporelings per cm of rope) to nets for removal to the open sea for cultivation. A month after culturing, the thalli reached a maximum length of 50 cm (average of 20–30 cm). The thalli grew faster as the water temperature dropped from 21°C to 17°C. It was proved that the vegetative cells isolated from a small thallus about 5 cm long could grow up into normal thalli after being kept frozen for a year. The results of this study show that vegetative cells can be used as new seeds to simplify the production of laver sporelings. This technique is a significant advance in the field ofPorphyra culture.     

东太平洋赤道公海茎柔鱼生物学特性

根据2013年4~6月在赤道公海探捕期间采集茎柔鱼样本,研究赤道公海茎柔鱼渔业生物学特性。结果表明,不同性别、不同月份,茎柔鱼胴长和体质量组成不同。雌、雄性样本的胴长范围分别为93~495 mm和94~406 mm,优势胴长组分别为260~340 mm和240~320 mm,分别占样本总数72.53%和70.26%;雌、雄性样本体质量范围分别为25~3 900 g和33~2170 g,优势体质量组分别为400~1 200 g和200~1 000 g,分别占样本总数82.71%和87.82%。雌、雄性茎柔鱼净体质量比例分别介于31.69%~80.51%和37.05%~81.35%之间,平均值分别为60.45%和62.73%。协方差分析表明,茎柔鱼胴长与体质量、胴长与净体质量的生长均存在性别间差异,胴长与体质量的生长都最适合用幂函数表示,而胴长与净体质量的生长,雌、雄性样本分别最适合用幂函数和指数函数表示。不同月份茎柔鱼性别比例构成不同,4月、5月和6月性别比例分别为1.87∶1、1.61∶1和1.17∶1,全部样本的性别比例为1.61∶1。不同月份茎柔鱼性腺成熟度的组成不同。不同性别、不同月份间茎柔鱼胃含物等级组成存在差异。     

盐度对菊黄东方鲀受精卵孵化和仔鱼生长的影响

比较在0、5、10、15、20、25、30(自然海水)、35、40、45、50等11个盐度梯度下,菊黄东方鲀(Takiugu flavidus)的受精卵发育和仔鱼生长情况,测定仔鱼的不投饵存活系数(SAI).结果表明:1)菊黄东方鲀受精卵在盐度0~50范围内均可孵化,最适盐度20~25.盐度(x)与孵化时间(y)呈正相关关系,表达式为:y=0.345 x+141.1,R2=0.956.2)实验盐度范围内,菊黄东方鲀仔鱼SAI值为1.55~30.10.3)在盐度5~45条件下,仔鱼皆可存活,存活率为3%~27%.其中,在10~30盐度下存活率较高,为20%~27%.经15 d的培育,仔鱼平均全长为6.28~7.34 mm.低盐度对菊黄东方鲀仔鱼生长更有利,在15~20盐度条件下,仔鱼生长较好,最终平均全长为7.30~7.34 mm.4)根据SAI值和生长状况,仔鱼生长和存活的适宜生长盐度为10~30,最适盐度范围为15~20.     

珠母贝人工苗养殖研究

在不同海区、不同水层养殖珠母贝人工苗,及用不同方法进行养殖小贝。结果表明:在开放性、半开放性、内湾性海区养殖11个月后,珠母贝壳高分别为64.5、51.4、69.8 mm,成活率分别为6.4%、0.8%、0.5%,其生长和成活率具有显著差异(P<0.05);同海区中3m、6m、9m、12m、15m水层养殖11个月后,珠母贝壳高分别为64.5、67.4、62.8、56.9、51.9mm,成活率分别为6.4%、6.8%、8.1%、11.7%、13.9%,随着水层的加深,珠母贝的生长变慢,成活率提高且差异显著(P<0.05);6个月龄小贝用穿耳法、片笼法、锥型笼法养殖5个月后,壳高分别长成56.1、55.7、51.7mm,成活率分别为72.6%、59.9%、51.7%,其生长和成活率具有显著差异(P<0.05)。     

Embryonic and larval development in barfin flounder Verasper moseri (Jordan and Gilbert)

Broodstock of Verasper moseri (Jordan and Gilbert) aged 3–4 years old were selected, and reinforced cultivation was conducted to promote maturation under controlled water temperature and photoperiod conditions. Fertilized eggs were obtained by artificial fertilization, and the development of embryos, larvae and juveniles was observed continuously. The results showed that the fertilized eggs of V. moseri were spherical, with transparent yolk and homogeneous bioplasm, and had no oil globule inside. The average diameter of the eggs was 1.77±0.02 mm. The eggs of V. moseri were buoyant in water with salinity above 35. The cleavage type was typical discoidal. Young pigment cells appeared when olfactory plates began to form. Hatching occurred at 187 h after fertilization at a water temperature of 8.5°C. The newly hatched larvae, floating on the water surface, were transparent with an average total length of 4.69±0.15 mm. During the cultivation period, when the water temperature was raised from 9 to 14.5°C, 4-day old larvae showed more melanophores on the body surface, making the larvae gray in color. The pectoral fins began to develop, which enabled the larvae to swim horizontally and in a lively manner. On days 7–8, the digestive duct formed. The yolk sac was small and black. The yolk sac was absorbed on day 11. Larvae took food actively, and body length and body height clearly increased. The rudiments of dorsal and anal fin pterygiophores were discernible and caudal fin ray elements formed on day 19. On day 24, the larval notochord flexed upwards, and the rays of unpaired fins began to differentiate. Pigment cells converged on the dorsal and anal fin rays, and the mastoid teeth on the mandible appeared. On day 29, the left eyes of juveniles began to move upwards. Depigmentation began in some juveniles and they became sandy brown in color on day 37. Most juveniles began to settle on the bottom of the tank. The left eyes of juveniles migrated completely to the right side on day 50, when the average body length attained 2.5±0.18 cm, and juveniles accomplished metamorphosis to young. The embryonic and larval characters of several flounder species are compared.     

Initiation of primary cell culture from amphioxus Branchiostoma belcheri tsingtauense

Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C.     

Laboratory observation on spawning,fecundity and larval development of amphioxus (Branchiostoma belcheri Tsingtaunese)

Although amphioxus is widespread in temperate and tropical seas, its population is diminishing because of environmental pollution. To keep the population of this evolutionarily important animal from diminishing, study on its reproduction and development is necessary. The main findings in this study on the spawning and fecundity of the amphioxus reared in laboratory and its larval development are as follows. 1. Water temperature markedly affected the spawning. It spawned only when water temperature reached 21°C. 2. Spawning of the amphioxus in laboratory was markedly extended. Initially, the amphioxus spawned at about 7:00 PM, but spawning time was postponed as spawning days went on. 3. The number of eggs produced by a female ranged from 1400 to 12800, average of 5800. This also represents the fecundity of the amphioxus because it shedded all eggs within the ovary at a time. 4. During the first few months of life of the amphioxus, its growth rate changed seasonally. The growth rate in summer and fall was greater than that in winter. 5. The pelagic larva became a benthic adult after 50 days. 6. The amphioxus reared in laboratory from fertilized eggs could produce fertile eggs and sperms. These findings can be a foundation for measures to address the problem of diminishing amphioxus population. Contribution No. 2274 from the Institute of Oceanology, Chinese Academy of Sciences This work was supported by CNNSF, and Chinese Academy of Sciences.     

Seasonal variation in abundance, diel vertical migration and body size of pelagic tunicate Salpa fusiformis in the Southern Yellow Sea

Mass occurrence of Salpa fusiformis was observed in the Southern Yellow Sea in May and June 2007. In order to investigate its population recruitment and environmental adaptation, temporal variation of abundance, diel vertical migration (DVM) and length frequency distribution of both aggregate and solitary forms were studied with samples collected from eight months during September 2006 to August 2007. S. fusiformis presented in six months other than September and October 2006, and average abundance of aggregate and solitary forms peaked in June and May, respectively. In December, aggregate forms were absent in the bottom layer and performed irregular DVM from surface to 50 m depth, while solitary forms was too scarce to perform diel vertical distribution analysis. Both aggregate and solitary forms presented reverse DVM in May and June. They migrated upwards during daytime and concentrated in surface layer at sunset. The bimodal distribution of aggregate forms was found in April and the average size was largest in this month. In other months, the smaller aggregate forms (1-5 mm) dominated in populations except for May, when the modal size ranged from 2 to 8 mm. The average size of solitary forms was largest in December, followed by April. The skewed nomal distribution of solitary forms was found in May and June, with the modal size of 2-7 mm and 5-13 mm, respectively.     

A Successful Microbound Diet for the Larval Culture of Chinese Shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselrnis chuii, rotifer （Brachionus plicatilis） and Arternia sp. Larvae of 0 to 13d post-hatch （dph） were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L^-1 in tanks, each containing 50 L sterilized seawater with salinity 30-32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13 dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13dph, which was not significantly different from that of larvae fed on live foods （63.55%）. The body length and development index （DI） of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13 dph （P〈 0.05）. The mean dry weight loss of the microbound diet was 9.2% after 90 min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 52.23% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

Initiation of primary cell culture from amphioxus <Emphasis Type="Italic">Branchiostoma belcheri tsingtauense</Emphasis>

Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C. Supported by Doctoral Initial Fund of Ludong University (No.43304)     

线纹尖塘鳢仔幼鱼的生长发育与池塘育苗研究

线纹尖塘鳢的胚后发育大致分为仔鱼、稚鱼、幼鱼和成鱼期。初孵仔鱼仅2.875mm。前期仔鱼混合营养期短,仅2～3d。后期仔鱼为器官发育、奇鳍条基本形成期,历时10d,平均全长从4.31mm增至8.97mm。稚鱼为器官分化完毕,奇偶鳍条分节、鳞被形成期,历时27～29d,长至平均全长27.79mm。早期幼鱼鳞被等发育完善,进入生长时期。仔鱼开口饵料为150～200μm轮虫,其摄饵大小与口宽呈正相关。15mm后稚鱼可驯饵摄食人工混合料。对仔、稚、幼鱼的生长测定结果显示,全长与日龄呈线性关系L=0.5289t-0.7354(r2=0.9720),体重与日龄呈指数函数关系,W=0.8236e0.1267t(r2=0.9611),全长与体重呈幂函数关系,曲线回归方程W=0.0045L3.2527,b=3.2527,属匀速生长型。采用池塘培水,添加外源饵料生物系列,稚鱼全长15mm时开始驯饵。2002-2005年共培育全长2.2～3.4cm早期幼鱼523.7万尾,仔鱼培育成15mm稚鱼的成活率为26.8%,培育成早期幼鱼的成活率为75.7%。     

A successful microbound diet for the larval culture of Chinese shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselmis chuii, rotifer (Brachionus plicatilis) and Artemia sp. Larvae of 0 to 13d post-hatch (dph) were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L⁻¹ in tanks, each containing 50 L sterilized seawater with salinity 30–32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13 dph, which was not significantly different from that of larvae fed on live foods (63.55% ). The body length and development index (DI) of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13dph (P<0.05). The mean dry weight loss of the microbound diet was 9.2% after 90min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 5223% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

美洲鲥仔稚鱼淡、海水培育试验

在室内水泥池用淡水和海水培育美洲鲥(Alosa sapidissima)仔鱼,研究两种培育模式下美洲鲥的生长。结果表明,美洲鲥仔稚鱼可较好地适应海水培育环境,经33 d的培育,淡、海水培育模式下美洲鲥平均全长日增长分别为0.965 7 mm和0.922 0 mm,前者略高于后者;成活率分别为70.3%和74.7%,海水培育模式成活率略高于淡水培育模式;仔稚鱼培育期间的生长模型方程分别为Y=9.614 3+0.500 7 X+1.326 6 X2和Y=9.478 3+0.558 4 X+1.038 0 X2(Y为全长,X为日龄)。可见,两种培育模式生长差异不显著,只要生产条件许可,均可采用淡水或海水培育美洲鲥苗种。     

Short-term morphological changes after sand-digging during a sand sculpture activity

The morphology of the beach backshore and foreshore at Huiquan Bay, Qingdao, China, is characterized by a single intertidal sandbar system with a spring tide range of 4.59 m. The beach was measured with a laser total station of Leica TPS402. Contours of the beach were generated using data collected in March and November 2005. The survey method provided 2 mm measuring accuracy and 4–10 m horizontal spacing. The net accretion volume of the foreshore was about 11, 215 m³ from March to November. After sand sculpture activity, the axis of the sand trough migrated onshore from about 3.5 m to 17.5 m on the foreshore beach in November. At the same time, the axis of the sandbar crest migrated onshore no more than 42.25 m on the northwest foreshore; and it migrated offshore no more than 23.75 m on the southeast foreshore. On the northwest and southeast foreshore beach, two strips of erosion areas with a thickness of 0–0.2 m appeared on the sandbar crest. Accretion occurred at the bottom of the sand trough with a thickness of ∼0.2–0.6 m. The sandbar height decreased after sand sculpture activity, and it was no more than 0.7 m in March and 0.6 m in November. Human activities, such as sand digging on the sandbar crest during sand sculpture activity, also can disturb the beach morphology of intertidal bar systems. This phenomenon also was validated by comparison of beach morphology, the results of a color artificial tracer experiment and a sediment transportation trend prediction.     

军曹鱼仔鱼期的摄食与生长

以人工孵化的军曹鱼(Rachycentron canadum)卵黄囊期仔鱼为材料,对仔鱼早期阶段的摄食及生长情况进行研究。结果表明:在水温28~32℃条件下,军曹鱼仔鱼3日龄开始摄食,6日龄时卵黄完全耗尽,6~7日龄饥饿仔鱼进入不可逆点(PNR)期;混合营养期3~4 d。饥饿仔鱼的初次摄食率开始较低,此后上升,到卵黄囊快要耗尽时达高峰(50%),以后又下降。内营养期,仔鱼日平均生长率为0.86 mm/d;PNR前的摄食期,饥饿仔鱼的日平均增长率为0.068 mm/d,而摄食仔鱼的日平均生长率为0.64 mm/d;PNR后至死亡前,饥饿仔鱼体长呈负增长,而摄食仔鱼的日平均生长率达1.04 mm/d。摄食仔鱼在孵化后8 d内的全长和日龄呈指数关系;前10 d的口径与全长呈正比关系。     

Breeding and mass scale rearing of clownfish Amphiprion percula: feeding and rearing in brackishwater

Breeding and mass scale larval rearing of clownfish Amphiprion percula is very limited in brackishwater. We designed an indoor program of A. percula culture in brackishwater with a salinity of 24±1, during which the impacts of feed type, water temperature, and light intensity, on the efficiency of its reproduction, were revealed. The fish were accommodated along with sea anemones in fibre glass tanks to determine the influence of brooder diet on breeding efficiency. Higher reproductive efficiency [number of eggs laid (276 22.3 eggs)] was observed when fish were fed live Acetes sp. rather than clam (204 16.4 eggs), trash fish (155 12 eggs) and formulated feed (110 10 eggs). The spawning rate was increased during September and October (water temperature, 28.74 0.55°C) on average of 2.4 spawning per month; and low spawning rate was in January (water temperature, 24.55 0.45°C) on average of 1 spawning per month. Among three light intensities (100, 500, and 900 lx) set to evaluate larval survival rate, larvae showed the highest survival rate (65.5%) at 900 lx. The breeding method specifically in brackishwater developed in the present study is a new approach, will help the people from the regions of estuary and backwater to enhance their livelihood and it will lead to reduce the exploitation from the wild habitat.     

罗非鱼开口饵料的初步研究

研究了配合饵料、轮虫、蛋黄、轮虫+人工配合饵料(3～12日龄投喂轮虫,13日龄起投喂配合饵料)等4种开口饵料对奥尼罗非鱼仔鱼生长、发育和存活影响。结果表明,1)投喂不同饵料对奥尼罗非鱼仔鱼生长具有显著影响(P<0.05),轮虫+配合饵料组生长较快,绝对增重率和特定增重率分别为0.011g/d和17.18%d-1;蛋黄组生长最慢,绝对增重率和特定增重率与其他各组差异显著(P<0.05)。2)轮虫+配合饵料与轮虫组的仔鱼成活率较高,分别为90.3%和89.7%,2组差异不显著(P>0.05),且仔鱼发育较好,大部分个体进入稚鱼阶段;蛋黄组成活率最低,只有40.7%,个体发育较为缓慢,与其他各组差异显著(P<0.05)。可见,投喂鲜活的天然饵料(轮虫)+配合饵料是罗非鱼苗种培育的最佳开口饵料。     

企鹅珍珠贝人工苗生长的初步观察

为了解企鹅珍珠贝生长规律，观察了壳高2.5～5.0mm的出池幼苗，按月测量其生长参数和成活率，以及环境因子。结果表明，企鹅珍珠贝生长最快的月份为7～11月和次年4～6月，壳长，壳高和壳宽月均增加量分别为3.8～13.0mm、4.7～11.2mm、2.3～3.8mm，月成活率97.8%～98.6%。企鹅珍珠贝生长最慢月份是11月至次年3月。     

Cryopreservation of Veliger Larvae of Trumpet Shell,Charonia sauliae: an Essential Preparation to Artificial Propagation

Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocol for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants (ethylene glycol, 1, 2-propanediol,dimethyl sulfoxide and glycerol) were employed at three concentrations (1.0, 1.5 and 2.0 molL-1) respectively and survival rates of larvae were determined after a storage of 1h. The larvae frozen with these four cryoprotectants after 1h storage were cultured, and then survival rates were determined at 24, 72 and 120h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL-1 showed the best protective effect in all experiments (p<0.05). And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1,7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL-1 dimethyl sulfoxide after 1h, 1d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34% ±11.28%, 83.10% ±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods (p>0.05).