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.     

The potential of ocean acidification on suppressing larval development in the Pacific oyster Crassostrea gigas and blood cockle Area inflata Reeve

We evaluated the effect of pH on larval development in larval Pacific oyster(Crassostrea gigas) and blood cockle(Arca inflata Reeve).The larvae were reared at pH 8.2(control),7.9,7.6,or 7.3beginning 30 min or 24 h post fertilization.Exposure to lower pH during early embryonic development inhibited larval shell formation in both species.Compared with the control,larvae took longer to reach the D-veliger stage when reared under pH 7.6 and 7.3.Exposure to lower pH immediately after fertilization resulted in significantly delayed shell formation in the Pacific oyster larvae at pH 7.3 and blood cockle larvae at pH 7.6 and 7.3.However,when exposure was delayed until 24 h post fertilization,shell formation was only inhibited in blood cockle larvae reared at pH 7.3.Thus,the early embryonic stages were more sensitive to acidified conditions.Our results suggest that ocean acidification will have an adverse effect on embryonic development in bivalves.Although the effects appear subtle,they may accumulate and lead to subsequent issues during later larval development.     

The potential of ocean acidification on suppressing larval development in the Pacific oyster Crassostrea gigas and blood cockle Arca inflata Reeve

We evaluated the effect of pH on larval development in larval Pacific oyster （Crassostrea gigas） and blood cockle （Arca inflata Reeve）. The larvae were reared at pH 8.2 （control）, 7.9, 7.6, or 7.3 beginning 30 min or 24 h post fertilization. Exposure to lower pH during early embryonic development inhibited larval shell formation in both species. Compared with the control, larvae took longer to reach the D-veliger stage when reared under pH 7.6 and 7.3. Exposure to lower pH immediately after fertilization resulted in significantly delayed shell formation in the Pacific oyster larvae at pH 7.3 and blood cockle larvae at pH 7.6 and 7.3. However, when exposure was delayed until 24 h post fertilization, shell formation was only inhibited in blood cockle larvae reared at pH 7.3. Thus, the early embryonic stages were more sensitive to acidified conditions. Our results suggest that ocean acidification will have an adverse effect on embryonic development in bivalves. Although the effects appear subtle, they may accumulate and lead to subsequent issues during later larval development.     

Effects of calanoid copepod Schmackeria poplesia as a live food on the growth, survival and fatty acid composition of larvae and juveniles of Japanese flounder, Paralichthys olivaceus

Zooplankton constitutes a major part of the diet for fish larvae in the marine food web, and it is generally believed that copepods can meet the nutritional requirements of fish larvae. In this study, calanoid copepod Schmackeria poplesia, rotifer Brachionus plicatilis and anostraca crustacean Artemia sp. were analyzed for fatty acid contents, and were used as live food for culturing larval Japanese flounder, Paralichthys olivaceus. The total content of three types of HUFAs (DHA, EPA and ARA) in S. poplesia was significantly higher than that in the other two live foods (P<0.01). Three live organisms were used for raising larvae and juveniles of Paralichthys olivaceus respectively for 15 and 10 d. Then the growth, survival and fatty acid composition of the larvae and juveniles were investigated. The results showed that the larvae and juveniles fed with copepods (S. poplesia) had significantly higher growth rate than those fed with the other two organisms (P<0.01). The survival of the flounder larvae fed with copepods was significantly higher than that of the others (P<0.01), and the survival of the juvenile fish fed with copepods was higher than that fed with Artemia (P<0.05). The contents of three types of HUFAs (DHA, EPA and ARA) and the ratio of DHA/EPA in larval and juvenile flounder P. olivaceus were analyzed. The results showed that the contents of DHA, EPA and ARA in the larvae and juveniles fed with S. poplesia were higher than those fed with a mixed diet or Artemia only, and the ratio of EPA/ARA in larvae and juveniles of P. olivaceus fed with S. poplesia was lower than that in the case of feeding with a mixed diet or Artemia only. The present data showed that copepod is the best choice for feeding the larvae and juveniles of fish considering its effects on the survival, growth and nutrition composition of the fish.     

Autumn larval fish assemblages in the northwest African Atlantic coastal zone

A study on the assemblage composition and vertical distribution of larval fish was conducted in the southern area of the Moroccan Atlantic coast in Autumn 2011. A total of 1 680 fish larvae taxa were identified from 21 families. The majority of the larvae were present in the upper layers. Clupeids were the most abundant larvae taxa followed by Myctophidae, Gadidae and Sparidae, hence the larval fish assemblages(LFA) were variable in different depth layers. Total fish larvae showed a preference for surface layers, and were mainly found above 75 m depth, with some exceptions. The maximum concentration of fish larvae was concentrated up to 25 m essentially above the thermocline, where chlorophyll a and mesozooplankton were abundant. Spatially, neritic families were located near the coast and at some offshore stations especially in the northern part, while oceanic families were more distributed towards offshore along the study area. Cluster analysis showed a segregation of two groups of larvae. However, a clear separation between neritic families and oceanic families was not found. Multivariate analysis highlighted the relationship between the distribution of larvae of different families and environmental parameters. Temperature and salinity seem to have been the factors that acted on associations of fish larvae. Day/night vertical distributions suggest there was not a very significant vertical migration, probably due to adequate light levels for feeding.     

The post-larval and juvenile fish assemblage in the Sukhothai Floodplain,Thailand

This study investigated abundance, species composition and spatial and temporal distributions of fish larvae and their relationship with some environmental variables in the Sukhothai floodplain in northern Thailand. Fish larvae were collected from 33 sampling stations on 8 occasions between August 2010 and October 2013. The study collected and identified 149 296 individuals, representing 32 families and 165 taxa. The species composition of larval fish was dominated by the Cyprinidae(47.27%), Cobitidae(7.88%), Siluridae(6.67%), Bagridae(6.06%) and Mastacembelidae(3.33%) families. The mostabundant larval species were the Striped flying barb Esomus metallicus(16.90%), the Siamese mud carpHenicorhynchus siamensis(8.48%) and the Sumatran river sprat Clupeichthys goniognathus(8.31%). The greatest abundance and species diversity of larvae were found when the river flow runs onto the floodplain. PCA and nMDS analysis revealed that the samples plot is associated with temporal distribution among years. The discharge was a major factor determining fish larvae assemblage and environmental variables in the Sukhothai floodplain. Four fish larval species were positively correlated with the samples for 2013. The result of the CCA ordination plot showed that only the discharge variable was strongly correlated with fish larvae abundance, especially two cyprinid R asbora species.     

Effect of n-3 HUFA levels in rotifers and Artemia on growth and survival of larval black sea bream （Sparus macrocephalus）

Requirement for dietary n-3 HUFA (n-3 highly unsaturated fatty acid) for growth and survival of black sea bream (Sparus macrocephalus) larvae was studied using rotifers and Artemia at various levels of n-3 HUFA. Five treatments with rotifers and Artemia differing in n-3 HUFA were prepared by enriching them with various oil emulsions. Results indicated that dietary n-3 HUFA significantly influence fish n-3 HUFA levels and are essential for growth and survival of black sea bream larvae. The results also indicated that the incorporation of n-3HUFA TG (triacylglycerols) into tissues of larval black sea bream was more effective from natural fish oil in comparison with n-3 HUFA fatty acid ethyl esters from ethyl-esterified oil.     

The critical period of high mortality of larvae fish —A discussion based on current research

Most fishes suffer high mortality during early development stages. The existence of a “critical period” was hypothesized by Hjort (1914). However, several decades of research have resulted in considerable controversy over the hypothesis. This paper attempts to clarify the critical period concept, demonstrate its usefulness, and resolve some of the controversy surrounding it. Based on our own studies and the literature, we find that within the “aquaculture school” (those concerned with culturing fishes, particulary larval stages), there is a great deal of evidence for the critical period concept. But we redefine the concept of “critical period” of mortality of fish larvae in this way. The deepest, sharpest, and fastest physiological and ecological changes during the development of the larvae are produced with the shift from endogenous nutrition to exogenous. At this stage, the old functions are quickly being replaced by new functions inside the body. Because of this shifting of function and imperfection of organs, the larvae are most sensitive at this time to environmental factors, particularly food supply. A higher specific mortality rate often occurs immediately folowing the period of strictly endogenous yolk feeding, and during the period of first exogenous feeding. Then it may be followed by a lower specific mortality rate. The significance of a “critical period” for fish populations is discussed. A good understanding of the “critical period” would provide a means for predicting the strength of year classes in natural fish populations, and also suggests approaches for increasing survival in aquaculture systems.     

Toxicity of organophosphorus pesticide sumithion on larval stages of stinging catfish <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>

Sumithion is widely used to control brittle in paddy fields and tiger bug in fish larval rearing ponds. The objective of this study was to elucidate the toxic effects of sumithion on larval stages of stinging catfish Heteropneustes fossilis. Larvae were exposed to two concentrations (150 and 250 μg/L) of sumithion with one control in three replicates of each. Larvae samples were collected at 20- and 24-h intervals followed by observation under a digital microscope. Exposures of stinging catfish larvae to sumithion produced deformities including irregular head shape, lordosis, yolk sac edema, body arcuation, tissue ulceration, etc. The mortality rates of larvae were significantly increased in response to increase in sumithion concentrations. Furthermore, around 30% of the total adult stinging catfish reared in sumithiontreated aquaculture ponds were found to be deformed permanently. These findings highlight that exposure of stinging catfish to sumithion at the critical and sensitive stages in their life cycle may significantly reduce the number of returning adults. Therefore, the use of sumithion for crop protection needs to be considered carefully and alternatives to sumithion should to be developed for controlling aquatic insects in aqua-ponds during larval rearing.     

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.     

光合细菌在水产养殖中应用的研究(3)——光合细菌作为仔鱼开口饵料的价值

本文所报导的是用光合细菌作为仔鱼开口饵料的效果。实验结果表明,单纯使用光合细菌,鱼(?)成活率较高,但因营养成份不全面,仔鱼生长速度较慢;如将光合细菌和其它饵料混合使用,则仔鱼在体长、体重和成活率方面均有显著提高。     

Phospholipid Compositions in Portunus trituberculatus Larvae at Different Developmental Stages

Phospholipids are used to improve the growth and survival of Portunus trituberculatus,a widely cultured crab species in China.However,only total phospholipids or several classes are applied in crab diets.In this study,we employed a targeted lipidomic method to investigate the comprehensive phospholipid composition in P.trituberculatus larvae and reveal the changing phospholipid profile over the larval development.Results showed that P.trituberculatus larvae contain 112 phospholipid species belonging to 10 phospholipid classes,in which phosphatidylcholine(PC)and phosphatidylethanolamine(PE)species are the most abundant,and PC,PE,phosphatidic acid,and phosphatidylserine(PS)are with high concentrations.The levels of all phospholipids significantly changed with larval development,which was highlighted by the downward parabolic changes in PE,phosphatidylglycerol,phosphatidylino-sitol,PS,lysophosphatidic acid,and sphingomyelin levels.In addition,nearly all phospholipid species were depleted at the M stage,which probably contributed to the mass mortality of crab larvae.These findings on the composition and alterations of phospholipids in P.trituberculatus larvae provide novel perspectives for the targeted supplementation of phospholipids in crab diets.Our work also highlights the use of targeted UHPLC-MS lipidomics in understanding the changes of phospholipids during crab development.     

Spatiotemporal distribution of fish eggs and larvae in the Huanghe(Yellow) River estuary,China in 2005–2016

Decreasing fish resources in estuaries is a subject of anthropogenic activities.Studies of the spatiotemporal distribution of fish eggs and larvae can help identify the status and processes underlying recruitment in a fishery.As the fifth largest river estuary in the world,the Huanghe(Yellow)River estuary(HRE)is a typical estuary that has been seriously affected by human activities.Annual surveys on ichthyoplankton and environmental factors were conducted in the months of May of 2005 and 2009-2016 in the HRE to investigate the spatiotemporal distribution of fish eggs and larvae and the associated influencing factors.A total of 23 and 20 species of eggs and larvae,respectively,were collected.The dominant orders were Perciformes(51.2%)and Clupeiformes(25.6%).The average number of fish species eggs and larvae were 6.0 and 4.1 in average abundance of 0.91 and 0.13 ind./m~3,respectively.The dominant species were mainly low-commercial-value small-sized fishes,such as Clupanodon punctatus,Hare,ngula zutnasi,and Acanthogobius,whereas certain traditional commercial fishes,such as Trichiurus lepturus,and Clupea pallasii,were not seen.Analysis of the fish egg and larval community revealed four temporal assemblages and two spatial assemblages.Salinity was the main factor on the spatial distribution of ichthyoplankton communities,the species number and Shannon-Weiner diversity index(H')of the fish egg and larval community near the river mouth with lower salinity were significantly lower than the community far away from the river mouth with higher salinity.In addition,increases of water temperature promoted the number and abundance of fish species eggs,and the areas of abundant prey tended to have a more diversified and abundant of ichthyoplankton species.In overall,overfishing,dam construction,and other human activities were the main drivers that led to the substantial decline in fishery resources in the HRE.     

Effect of n?3 HUFA levels in rotifers and Artemia on growth and survival of larval black sea bream ( Sparus macrocephalus )

Requirement for dietary n−3 HUFA (n−3 highly unsaturated fatty acid) for growth and survival of black sea bream (Sparus macrocephalus) larvae was studied using rotifers andArtemia at various levels of n−3 HUFA. Five treatments with rotifers andArtemia differing in n−3 HUFA were prepared by enriching them with various oil emulsions. Results indicated that dietary n−3 HUFA significantly influence fish n−3 HUFA levels and are essential for growth and survival of black sea bream larvae. The results also indicated that the incorporation of n−3HUFA TG (triacylglycerols) into tissues of larval black sea bream was more effective from natural fish oil in comparison with n−3 HUFA fatty acid ethyl esters from ethyl-esterified oil. Project No. 39770588 supported by the NSFC.     

Effect of zinc on the growth and development of larvae of bay scallopArgopecten Irradians

The 48-h straight-hinge larvae ofArgopecten irradians were exposed for 9 days to various concentrations of zinc in seawater. The growth rate of larvae in the 50 ppb Zn mediums was 77% that of the controls, and nearly zero in the 200 ppb Zn mediums. A progressive decrease in growth and increase in larval deformity and mortality was observed with increasing zinc concentrations from 50 to 200 ppb. 50 and 100 ppb Zn resulted in delayed development of eye spot and metamorphosis and reduced numbers of larvae at both stages. All larvae subjected to higher zinc concentrations died before attaining the eye-spot stage. The zinc concentrations which caused 50% reduction in growth (G C₅₀) and 50% mortality (LC₅₀) were 109 and 120 ppb respectively. Growing in the medium with 100 ppb Zn, the larvae fed withIsochrysis galbana cultured in a medium with 100 ppb Zn showed more suppressed growth and development than those fed with normal food. It is suggested that zinc contaminated food has strong influence on the bay scallop larvae. Contribution No. 1616 from the Institute of Oceanology, Academia Sinica.     

Effects of Seawater Acidification on Early Development of Clam Cyclina sinensis

Anthropogenic emission of atmospheric carbon dioxide(CO_2) has led to a rapid increase in atmospheric CO_2 concentration. Increasing atmospheric CO_2 can reduce seawater pH and carbonate ions, which may adversely affect the survival of the larvae of calcareous animals. Cyclina sinensis is a commercially and ecologically important species in several Asian countries. Living in coast shallow waters, this species has experienced the coastal environmental changes frequently throughout its life cycle. In this study, we simulated possible future seawater pH values including 8.2, 7.8 and 7.4 and examined the effects of ocean acidification on the early development of C. sinensis. Clam embryos were incubated for 48 h(2 d) in control and high-CO_2 seawater to compare embryogenesis, larval growth and swimming behavior. Fertilization rate was quite sensitive to pH, and moderate acidification could induce a significant decrease in fertilization rate. However, only extreme acidification could bring significant negative effect to hatching rate, body size, and average path velocity of trochophora. Moreover, with seawater acidification, C. sinensis needs much more time to reach the same developmental stage, which increases the risk of larva survival. Together with recent studies demonstrating negative impacts of high CO_2 on fertilization and larva swimming behavior, the results imply a future decrease of C. sinensis populations in oceans if its acclimation to the predicted environmental alteration does not occur.     

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。     

Effects of base analogues 5-bromouracil and 6-aminopurine on development of zebrafish Danio Rerio

Zebrafish （Danio rerio） genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are reported for the first time in this study. The early development of the zebrafish embryos was little affected by 5-bromouracil and 6-aminopurine, while the late development （organogenesis） was significantly impaired. Embryos exposed to 5-bromouracil mainly showed curled tail, wavy body, golden pigmentation and the mouth with protruding lower jaw. 6-aminopurine-treated embryos had defective anterior structures, curled tails and wavy body. RAPD analysis showed that the majority of 5-bromouracil-and 6-aminopurine-treated larvae and fish shared banding patterns in common with the control, suggesting that most mutagenesis induced by these agents are point mutations. However, some fish derived from 5-bromouracil-treated embryos had golden（gol） pigmentation; and RAPD analysis revealed that their band patterns differed from those of the control.Possibly, 5-bromouracil can occasionally cause relatively extensive changes in the fish genome. The results of this study may provide valuable help for detailed studies of mutagenesis.     

Effects of elevated seawater pCO<Subscript>2</Subscript> on early development of scallop <Emphasis Type="Italic">Argopecten irradias</Emphasis> (Lamarck, 1819)

To investigate the effects of elevated seawater pCO₂ on the early developmental stages of marine benthic calcifying organisms, we exposed the eggs and larvae of Argopecten irradias, an important bivalve species in Chinese aquaculture, in seawater equilibrated with CO₂-enriched (1000 ppm) gas mixtures. We demonstrated that elevated seawater pCO₂ significantly interfered with fertilization and larval development and resulted in an increased aberration rate. Fertilization in the treatment (pH 7.6) was 74.3% ± 3.8%, which was 9.7% lower than that in the control (pH 8.3) (84.0% ±3.0%). Hatching success decreased by 23.7%, and aberration rate increased by 30.3% under acidic condition. Larvae in acidified seawater still developed a shell during the post-embryonic phase. However, the shell length and height in the treatment were smaller than those in the control. The development of embryos differed significantly at 12 h after fertilization between the two experimental groups. Embryos developed slower in acidified seawater. Nearly half of the embryos in the control developed into D-shaped larvae at 48 h after fertilization, which were considerably more than those in the treatment (11.7%). Results suggest that future ocean acidification (OA) would cause detrimental effects on the early development of A. irradias.