A method to determine protein digestibility of microdiets for larval and early juvenile fish

A method to evaluate protein quality using in vivo methods was developed for larval fish. FluoSpheres^® fluorescent microspheres (10 μm) were incorporated into two test diets, our standard zein microdiet (ZMD) and a microdiet with identical ingredients except for the replacement of high quality fish meal with the same product cooked for 24 h at 80 °C (ZMD-CF). Several trials were performed to design a reliable method to test digestibility using FluoSpheres^® as a marker. The developed in vivo technique was tested on 35 days posthatch (dph) larval Atlantic cod ( Gadus morhua L.) and two tropical fish species in the early juvenile stage. The method took into account loss of total protein to the faecal pellet and water column. Apparent digestibility of protein in larval cod fed ZMD was significantly higher than that of larvae fed ZMD-CF ( P  < 0.05). A growth study to validate differences between the two diets showed significant differences in growth and survival of larvae fed ZMD versus ZMD-CF ( P  < 0.05). Further validation of our results was indicated through the use of a pH-stat method using enzymes extracted from 35 dph larval cod guts. This novel technique will be advantageous for researchers to evaluate feed ingredients for larval marine fish and is adaptable to many different areas of larval fish nutrition.     

Immunostimulatory effects of dietary poly‐β‐hydroxybutyrate in European sea bass postlarvae

The stable production of high‐quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly‐β‐hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In this study, the effect of feeding PHB‐enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) postlarvae was examined. Amorphous PHB was administered to 28‐day‐old sea bass postlarvae over a period of 10 days. The survival and growth performance were monitored, and the expression of 29 genes involved in immunity, growth, metabolism and stress‐response was measured. While the expression of the insulin‐like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed postlarvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies.     

五种重金属对早繁鮸鱼胚胎和仔鱼的毒性效应

在水温27.5℃、盐度29条件下,研究了Cu2+、Cd2+、Zn2+、Cr6+和Pb2+5种重金属对早繁鮸鱼胚胎发育和初孵仔鱼的影响。结果表明:重金属在胚胎发育过程中引起各种畸形,如胚胎尾巴弯曲、初孵仔鱼不能出膜、胚胎死亡等;胚胎和初孵仔鱼上出现黑色颗粒,初孵仔鱼脊柱弯曲成L、V、S形或尾部没有展开呈逗号状,褶皱边缘被破坏等畸形。Cu2+、Cd2+、Zn2+、Cr6+和Pb2+5种重金属均延迟胚胎发育速率。综合孵化率、胚胎畸形和仔鱼畸形率等指标得出5种重金属对胚胎的毒性大小依次为Cu2+>Cd2+>Zn2+>Pb2+>Cr6+。计算了5种重金属对早繁鮸鱼初孵仔鱼的LC50和安全浓度,得出5种重金属对早繁鮸鱼初孵仔鱼毒性大小顺序为:Cu2+>Cd2+>Pb2+>Zn2+>Cr6+。     

Improved performance of gilthead sea bream, Sparus aurata, larvae after ozone disinfection of the eggs

Ozone (O₃) dissolved in seawater (ODS) was evaluated, as an egg disinfectant, on the spawn of captive gilthead sea bream, Sparus aurata, brood stock. Four contact times (CT) were tested (0.6, 1.2, 2.4 and 4.8 mg min L⁻¹) where CT was calculated by multiplying the dissolved O₃ concentration (0.3 mg L⁻¹) by different exposure periods (2, 4, 8, 16 min). There was also a disinfected seawater treatment that contained no O₃ or derived compounds (CT 0) and an untreated seawater control. All ODS treatments reduced egg surface bacterial counts to zero, which was significantly (P<0.05) lower than the CT 0 and the control groups (194 and 1320 plate⁻¹ respectively). Nevertheless, the hatching rate was high in the control and the CT treatments 0, 0.6 and 1.2 (88.7%, 87.3%, 89.5% and 83.7% respectively) while eggs exposed to a CT 2.4 and 4.8 hatched poorly (36.5% and 20.4% respectively), which was likely due, at least in part, to larvae unable to break the egg chorion successfully. Swim‐bladder inflation was significantly higher in the ODS groups (>97%) compared with the control and CT 0 treatments (ca. 70%). The results suggest that a 2‐min exposure of eggs to 0.3 mg O₃ L⁻¹ of ODS (CT 0.6) would improve current protocols in marine larviculture.     

温度和盐度对鞍带石斑鱼受精卵发育及仔鱼成活率的影响

研究了在8个盐度梯度(18.3‰、19.7‰、21.2‰、23.5‰、25.6‰、27.1‰、29.5‰和31.7‰)下鞍带石斑鱼Epinephelus lanceol受精卵和仔鱼在水中的分布状态、出膜率和仔鱼存活率;在9个温度梯度(18℃、20℃、22℃、24℃、26℃、28℃、30℃、33℃和35℃)下,其受精卵孵化率、畸形率和仔鱼成活率。结果表明:(1)在水温为25~26℃,当盐度≤25.6‰时,受精卵表现为沉性;当盐度≥27.1‰时,受精卵表现为浮性。仔鱼的分布与受精卵有相似的特点,当盐度≤21.2‰时仔鱼分布在底层水体;当盐度≥27.1‰时,仔鱼集中分布在上层水体。(2)以孵化率和成活率为指标,鞍带石斑鱼孵化的适宜盐度范围为21.2‰~31.7‰,最适盐度范围为25.6‰~31.7‰。在盐度为31.7‰时仔鱼成活率最高达到75.9%。(3)鞍带石斑鱼胚胎发育的适温范围是26~30℃。28℃时受精卵孵化率、仔鱼成活率均最高,畸形率最低。     

大亚湾鱼卵、仔稚鱼种群特征与环境因子的相关关系

为了解大亚湾鱼卵、稚仔鱼种群动态变化及其与环境因子的关系, 2015年采用浅水I型浮游生物拖网对大亚湾鱼卵、仔稚鱼进行了4个季度的调查,结果表明,4个季度共采获鱼卵31361粒,仔稚鱼244尾。经鉴定分析共有27种,其中鱼卵19种,隶属于6目16科18属;仔稚鱼18种,隶属于5目15科16属;鱼卵、仔稚鱼共有种11个。鱼卵平均丰度为614.55 ind/1000 m^3,稚仔鱼的平均丰度为3.69 ind/1000 m^3。鱼卵丰度季节变化由高到低依次为夏季、春季、冬季、秋季;空间分布显示,丰度由高到低依次为中央列岛、人工鱼礁区、湾口东岸。鱼卵多样性指数(H￠)范围为0.05～2.26,平均值为0.86,稚仔鱼多样性指数(H?)范围为0～2.53,平均值丰度0.82;鱼卵均匀度指数(J)范围为0.02～0.99,平均值为0.50;稚仔鱼均匀度指数(J)范围为0～1.00,平均值为0.52。根据Pearson相关性结果分析表明,与种群关系最密切的环境因子为温度和溶解氧(P<0.01)。研究结果显示,春夏季是大亚湾海域鱼类产卵的重要时期,中央列岛海域是大亚湾鱼类繁殖的关键区域。     

缢蛏规模化苗种培育技术报告

利用室内工厂化育苗设施对缢蛏Sinonovacula constricta (Lamarck)进行人工催产,幼虫培育,盐度调控,育苗中后期直接投喂土池中天然藻类,减轻藻类培育负担,同步进行多批次苗种生产,大幅度增加了产量.2011年规模化苗种培育孵化率62.2％ ～ 70.7％,附着率52.2％ ～77.1％,幼虫成活率30.8％～ 60.2％,幼虫增重率为99.85％～99.96％,3个月内共催产915 kg种蛏,培育优质蛏苗约40亿粒.     

Ontogenetic development of digestive enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.