发酵豆粕对圆斑星鲽生长及表观消化率的影响

为探讨圆斑星鲽(Verasper variegatus)饲料中发酵豆粕的适宜添加量,本试验以0%、10%、20%、30%、40%的发酵豆粕添加在五组等蛋等脂的饲料中,分别替代鱼粉蛋白的0%,9.81%,19.63%,29.44%,39.26%,标记为A、B、C、D、E共5个试验组,每组设3个重复,每池15尾圆斑星鲽(49.0±1.0 g),经过10周的饲养试验,结果表明:随饲料中发酵豆粕水平的升高,特定生长率逐渐降低,A、B组显著高于其他各组(P<0.05),饲料系数则逐渐升高,A、B和C组显著低于D、E组(P<0.05)。营养物质表观消化率以B组最高,显著高于其他各组。综上所述,饲料中添加10%的发酵豆粕在一定程度上能提高鱼体对营养物质的表观消化率,且不会对鱼体生长造成不利影响,过量则抑制鱼体生长。     

Pedigree analysis of recaptured fish in the stock enhancement program of spotted halibut Verasper variegatus

ABSTRACT:   Spotted halibut Verasper variegatus hatchery juveniles produced in 2002 were genotyped using three microsatellite DNA markers (msDNA) and then released into natural waters. Subsequently, recaptured individuals were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, from the genetic point of view, a pairwise F _ST test was implemented to estimate the genetic divergence between the wild captive broodstock, the hatchery offspring and the recaptured samples. The analysis showed significant differentiation between the broodstock and recaptured samples. Pedigree determination using msDNA was used to calculate the effective population size of the recaptured stock, which was found to be very low ( N _e ≈ 8). Equal family survivability was observed between the two recaptured stocks, but not between the released and recaptured stock. The number of identified families was higher and more equalized in the hatchery offspring compared to the recaptured samples, where the number of families declined. This fact was caused by an unequal family survivability just before or just after release. Separately, the number of contributing parents to the hatchery offspring was lower than the broodstock census number. Consequently, these two facts caused the genetic divergence of the recaptured stock from the broodstock.     

双酚A和壬基酚长期暴露对斑马鱼繁殖的影响

内分泌干扰物的研究目前正逐渐成为国际研究新的热点，但长期暴露条件下对生殖情况的研究目前较少。本文研究了环境类雌激素双酚A（BPA）以及和壬基酚（NP）联合的作用，对暴露一个世代的斑马鱼（F1）（Danio rerio）的生殖情况、子代质量的影响。将健康的受精卵48h后暴露于200，500，1000μg·L^-1 BPA以及三个浓度与30μg·L^-1 NP的联合，同时设置空白对照和30μg·L^-1 NP对照。130dph将成鱼置于清水喂养20d，统计每天的产卵量、产卵次数、畸形率、孵化率、卵径、破膜时间以及耐饥时间。以此为终点评估暴露后斑马鱼的生殖能力和子代质量。与对照相比NP，BPA可以抑制斑马鱼的生殖，两种类雌激素联合作用可以极显著地抑制斑马鱼的生殖（P〈0．01）；BPA各浓度组导致F1代畸形率上升（P〈0．05）；高浓度BPA抑制产卵量（P〈0．05）；BPA200，500μg·L^-1会延长F1代的破膜时间和耐饥时间（P〈0．01），而BPA1000μg·L^-1则产生相反的效果（P〈0．05）。双酚A对斑马鱼的生殖有明显的抑制作用，双酚A和壬基酚有协同效应。长时间暴露于双酚A会影响斑马鱼的生殖能力和子代质量。     

Growth and habitat of spotted halibut Verasper variegatus in the shallow coastal nursery area, Shimabara Peninsula in Ariake Bay, Japan

ABSTRACT:   To determine the growth performances and habitat characteristics of spotted halibut Verasper variegatus , successive samplings were conducted in the south-eastern coastal area of Shimabara Peninsula in Ariake Bay. The push-net samplings were performed at Kamaga beach from May 2003 to April 2004 to determine the horizontal distribution of larvae and juveniles, while the gill net samplings were done in the coastal waters of Tatsuishi beach from July 2003 to July 2004 to collect the larger juveniles and adults. A total of 478 individuals from metamorphosing larvae to 2-year-old fish (15.2–447.0 mm total length [TL]) were sampled. In particular, the 2003-year class fish were successively collected from May 2003 to June 2004 (15.2–350.0 mm TL; n  = 418) and the mean total length in March, June, September, December 2003 and May 2004 was 22.4, 82.5, 172.5, 203.9 and 296.4 mm, respectively. The present study found that metamorphosing larvae migrated into the estuarine tidal flats during March 2003; newly settled juveniles inhabited the shallow intertidal areas; and juveniles had a remarkably high growth in shallow coastal areas. It is suggested that young fish around 300 mm TL migrated from the nursery grounds in Ariake Bay to deeper waters around Tachibana Bay during May–June 2004.     

Genetic population evaluation of two closely related flatfish species, the rare barfin flounder and spotted halibut, along the Japanese coast

ABSTRACT:   Barfin flounder and spotted halibut have been selected as target species for stock enhancement in Japan. Understanding the genetic condition of the wild stock is a principal requirement in any stock enhancement program. The genetic variability of barfin flounder and spotted halibut, and the population structure of spotted halibut were evaluated using microsatellite DNA markers (msDNA) and the control region of the mitocondrial DNA (mtDNA). Barfin flounder and spotted halibut showed high genetic variability at the msDNA level. Barfin flounder A was 16.7 and H _e was 0.860; spotted halibut A _n ranged from 7.7 to 10.2 and H _e ranged from 0.710 to 0.774. At the mtDNA level, high haplotype ( h  = 0.922) and low nucleotide (π = 0.002) diversities were observed for barfin flounder; however, low haplotype and nucleotide diversities ( h  = 0.603–0.620 and π = 0.001–0.002), and very low haplotype and nucleotide diversities ( h  = 0.193 and π = 0.0003) were observed for spotted halibut in the north and south locations, respectively. Slight genetic differentiation among spotted halibut sampling locations was observed from the msDNA. MtDNA analyses showed genetic differentiation between north and south locations, but not within them. The designation of north-specific and south-specific management units in the future stock enhancement activities of spotted halibut is recommended.     

A comparison of retinol, retinal and retinyl ester concentrations in larvae of Atlantic halibut (Hippoglossus hippoglossus L.) fed Artemia or zooplankton

Atlantic halibut (Hippoglossus hippoglossus L.) larvae were fed enriched Artemia or zooplankton in duplicate tanks from 0 to 60 days after first‐feeding. Both diets and the larvae were analysed for vitamin A (VA) in order to confirm earlier findings, in which Artemia fed larvae had lower levels of VA compared with larvae fed zooplankton. Furthermore, we wanted to investigate the composition of the retinoids in the larvae. The results showed that Artemia and zooplankton contains low levels of VA, probably too low to sustain the assumed requirement. Nevertheless, larvae fed Artemia had the same level of retinal and retinol as larvae fed zooplankton. We found a significant lower level of retinyl esters in larvae fed Artemia. The total VA level was lower in larvae fed Artemia only at the end of the feeding trial after the onset of metamorphosis. Our conclusion is that feeding Artemia to Atlantic halibut larvae is not likely to cause VA deficiency.     

A correlation between phototactic response and first-feeding of Atlantic halibut (Hippoglossus hippoglossus L.) larvae

The correlation between positive phototaxis and feeding incidence at first‐feeding for groups of Atlantic halibut larvae ranging in age from 210 to 240 degreedays, post hatch, was examined. Phototactic response was measured as the fraction of larvae that responded by horizontal swimming towards a light source, and the median travel distance for the responding fraction of larvae. Within the ranges investigated, larval age, size or proportion of deformed larvae had no significant effect on the phototactic response, or on the feeding incidence (deformed larvae excluded from analysis). The fraction of larvae responding phototactically and the median travelling distance for responding larvae were significantly correlated. Feeding incidences after 24 and 48 h were also significantly correlated with both measures of phototactic response. The possibility of using phototactic response in quality assessment is discussed.     

Folate in eggs and developing larvae of Atlantic halibut, Hippoglossus hippoglossus L.

Folate mobilization from the yolk compartment during larval development was studied by analysing the folate concentration in whole body, embryo and yolk in a single batch of Atlantic halibut, Hippoglossus hippoglossus L., eggs and larvae that showed successful fertilization and development. There was a net loss of approx. 50% of folate from yolk during endogenous feeding. Further, only 23% of the decrease in yolk folate was retained in the larval body. The data suggest a need for folate for metabolic and growth purposes during embryogenesis of approximately 2 μg g^?1 weight gain. Relative to these data and published folate requirement for cold‐water species, batches of egg from 16 Atlantic halibut brood fish contained variable and, for some batches, critically low levels of folate. This may constitute a potential problem for larval development until start feeding.     

Growth, feed conversion efficiency and growth heterogeneity in Atlantic halibut (Hippoglossus hippoglossus) reared at three different photoperiods

Juvenile Atlantic halibut, Hippoglossus hippoglossus (initial weight (SD) 191.3 (±44.7)) g, were reared for 99 days at a constant temperature of 11°C and subjected to three different light regimes from 13 September to 21 December: continuous light (LD24:0), simulated natural photoperiod of Bergen (60°25′N, LDN) and constant 20 h light:4 h dark (LD20:4). The fish reared on the different photoperiod regimes differed in their growth patterns as juveniles exposed to long days, i.e. LD20:4 and LD24:0, exhibited faster growth than those experiencing a natural photoperiod. The LD20:4 group showed the highest average specific growth rate (0.72% body weight day^?1), whereas fish on LDN displayed the lowest average specific growth rate (0.60% body weight day^?1). The final mean weights of the LD20:4 and the LD24:0 groups were 15% and 12% higher than those of the LDN group. Dividing the duration of the experiment into three time periods shows that the LD20:4 and LD24:0 had a higher feed conversion efficiency (FCE) as compared with the LDN group during the first and the last period, while a reversed situation was observed in the second period. Our data indicate a larger variation in growth rates among individuals in the best‐performing groups (here LD20:4 and LD24:0). This may indicate that formation of size hierarchies is more pronounced in groups with more homogenous growth (here LDN). Overall, our findings indicate that extended light regimes result in faster growth and better feed conversion in juvenile Atlantic halibut. In line with findings on other flatfish species, this supports the concept that constant long day:short night or continuous light regimes should be used by the farmer in order to maximize growth and improve feed conversion in Atlantic halibut.     

Modelling the advection and diffusion of eggs and larvae of Greenland halibut (Reinhardtius hippoglossoides) in the north-east Arctic

Since the late 1980s there has been considerable uncertainty in recruitment levels of the north‐east Arctic stock of Greenland halibut (Reinhardtius hippoglossoides). The abundance of several year classes, originally considered very low at 0–3 yr age, appeared higher than expected at the age of 6+. This may be due to poor targeting of recruitment surveys of the younger year classes. The present work considers the transport and dispersion of eggs and larvae of Greenland halibut by numerical modelling in order to predict the locations of the initial recruitment grounds. Current fields from a 3D baroclinic hydrodynamic model are fed into a Lagrangian particle‐tracking model developed for the Barents Sea area. The particles are released into the current at the spawning field along the shelf slope from Lofoten to Bear Island (69–75°N). Vertically, the particles can follow a predefined depth‐by‐age curve or be kept at a fixed depth. This model system is used for different years to examine changes in the drift pattern. The results indicate that spawning location, transport depth and inflowing activity to the Barents Sea are important factors influencing the distribution of juveniles.     

Interactive effect of photoperiod and temperature on the growth rates,muscle growth and feed intake in juvenile Atlantic halibut

To investigate the interactive effects of temperature and photoperiod on the growth performance, feeding parameters and muscle growth dynamics in juvenile Atlantic halibut (Hippoglossus hippoglossus L.), a total of 1212 juvenile halibut, including 383 tagged fish (mean initial weight of tagged individuals: 17.6 ± 0.3 g SE), were reared under a simulated natural light regime for Bergen (60°25′N) or continuous light at 9, 12 and 15 °C from 3 December 2007 until 11 March 2008. The mean weight and growth rate were significantly higher at 12 and 15 °C than at 9 °C. In addition, significantly higher mean weight and growth rate were observed in halibut reared under continuous light at a low temperature, indicating an interactive effect of temperature and photoperiod on growth performance. No effect of temperature or photoperiod was found with respect to feed conversion efficiency, whereas a higher feed consumption at increasing temperature and a higher overall daily feeding rate at continuous light at a low temperature were observed. Indications of continuous light having a stronger effect at low temperatures on muscle growth dynamics were found. A difference in the size class distribution of fibre diameter was found between photoperiod treatments at 9 °C, suggesting that continuous light resulted in elevated hypertrophic growth at low temperature. This may suggest that the increased growth rate found at continuous light at 9 °C may be a result of hypertrophic growth in juvenile halibut.     

Control of light condition affects the feeding regime and enables successful eye migration in Atlantic halibut juveniles

Incomplete eye migration is one of the major problems in intensive production of juvenile Atlantic halibut. More than 60% of an average juvenile population reared according to best practice suffers from this abnormality. In commercial production, these fish are discharged and represent a substantial economic loss and a large welfare problem. In the present investigation it is demonstrated that by controlling diurnal light and darkness periods together with a meal based feeding regime, incomplete eye migration can be dramatically reduced in production systems for Atlantic halibut.Control groups were reared under continuous light conditions, whereas the experimental groups were given 7 h of darkness and 17 h of light during a 24 hour cycle, in a period lasting from 12 to 35 days post first-feeding. Otherwise both groups were reared under continuous light conditions. All larvae were fed short time enriched Artemia supplied two times daily.The experimental conditions did not affect the overall growth or survival up to day 85 after first feeding. However, 27 ± 3% of the fry reared under continuous light conditions had complete eye migration, whereas in juveniles reared under shifting light and darkness conditions, complete eye migration was 85 ± 7%. These results represent a major improvement in production systems for Atlantic halibut juveniles.     

The relation between age at first maturity and growth in Atlantic halibut (Hippoglossus hippoglossus) reared at four different light regimes

The effect of extended photoperiods on growth and age at first maturity was investigated in 166 (79 females and 87 males) individually tagged Atlantic halibut. The halibut were reared at 11°C on four different light regimes from 10 February to 6 July 1996: simulated natural photoperiod, (LDN), continuous light (LD24:0), constant 8 h light and 16 h darkness (LD8:16) and LD8:16 switched to continuous light on 4 May 1996 (LD8:16–24:0). From 6 July 1996 to 9 February 1998 the LD24:0 and LD8:16–24:0 were reared together under continuous light and the LDN and LD8:16 together under natural photoperiod. Juveniles subjected to continuous light exhibited faster growth than those experiencing a natural photoperiod or a constant short day. Moreover, the results suggest an overall growth enhancing effect of continuous light in females, but not in males. No females matured during the trial, but the proportion of mature males differed between the photoperiod groups, with significantly fewer males maturing in groups reared at continuous light. Independent of photoperiod regime and maturation status, females were significantly bigger than males from 14 April 1997 onwards. Immature males were bigger than maturing males from 23 March 1996 onwards. As continuous light reduced maturation at age 2+ in males, this could be used to reduce precocious maturation in males.     

The influence of water temperature on spawning patterns and egg quality in the Atlantic halibut (Hippoglossus hippoglossus L.)

A three-year study was conducted to investigate the effects of water temperature on Atlantic halibut broodstock reproductive performance. Two groups of fish held under ambient photoperiod were established onto contrasting temperature regimes. The ‘chilled’ group were held at below 9 °C from late October and at approximately 6 °C from December until the end of the spawning period whilst the ‘ambient’ group received no temperature control. The temperature profiles for the ‘ambient’ group changed over the 3 years but were generally characterised by warmer conditions prior to spawning and an earlier temperature rise in the spring. Total egg production was higher in the ‘chilled’ group each year. Absolute fecundity was significantly reduced in the ‘ambient’ group compared to the ‘chilled’ group every year of the study (0.6 million eggs/female for ‘chilled’ group vs. 0.3 to 0.4 million eggs/female for ‘ambient’ group) and egg viability, in terms of fertilisation and hatch rate, was significantly impaired in the ‘ambient’ group in years 2 and 3 (mean fertilisation rate in the ‘ambient’ group was between 27.0% and 54.8%; vs. 63.4% to 77.4% in the ‘chilled’ group, mean hatch rate in the ‘ambient’ group was between 3.1% and 25.6% vs. 60.7% to 71.7% in the ‘chilled’ group). Eggs spawned at high temperatures, later in the season were generally of low viability. In the ‘ambient’ group the spawning season became progressively delayed during the study and average duration of spawning season over 3 years was shorter (between 23.5 to 26.3 days for the ‘ambient’ group vs. between 30.5 and 41.2 days for ‘chilled’ group). It is hypothesised that high temperature during the vitellogenesis period caused a delay in spawning and a reduction in quantity and quality of eggs and that this effect was exacerbated by high temperature during spawning.     

圆斑星鲽促性腺激素释放激素基因克隆及表达特性

采用RACE技术,在圆斑星鲽(Verasper variegatus)脑中克隆到3种GnRH基因的cDNA序列: cGnRH-Ⅱ、sGnRH和sbGnRH.每种GnRH都包括1个信号肽、1个Gly-Lys-Arg连接序列和1个GnRH相关肽.其中, cGnRH-Ⅱ的cDNA全长为568 bp,编码85个氨基酸, ORF为255 bp,5′UTR为141 bp,3′UTR为169 bp.sGnRH的cDNA全长为457 bp,编码90个氨基酸, ORF为270 bp,5′UTR为41 bp,3′UTR为143 bp.sbGnRH的cDNA全长为381 bp,编码98个氨基酸, ORF为294 bp,5′UTR为48 bp,3′UTR为36 bp.分析了3种GnRH基因的编码氨基酸序列与其他脊椎动物的同源性,圆斑星鲽 GnRH 与鲽形目鱼类氨基酸同源性最高,其次为鲈形目鱼类.对3种 GnRH 基因的系统进化分析表明,圆斑星鲽 GnRH 基因与其他鲽形目鱼类亲缘关系最近,其次为鲈形目、鲑形目和鳗鲡目鱼类.荧光定量PCR分析表明,3种GnRH基因都在脑中表现出最高表达水平且具有性别特异性表达模式:雌性中不同组织的 GnRH mRNA 表达水平都相应高于雄性.组织表达分析表明, cGnRH-Ⅱ的 mRNA 仅在脑中表达,而sbGnRH在各个组织都有表达, sGnRH仅在脑、垂体和性腺中表达.脑中sbGnRH mRNA的表达水平在卵巢成熟过程中变化显著(P<0.05),而其他两种GnRH的mRNA表达水平变化不显著(P>0.05).本研究首次在圆斑星鲽脑中克隆到3种GnRH基因,其组织和季节表达水平变化表明sbGnRH可能是圆斑星鲽生殖调控的关键GnRH类型,本结果可为圆斑星鲽生殖调控机制和人工繁育技术研究提供理论支撑.