Post-stocking dispersal of European grayling,Thymallus thymallus (L.), in a semi-natural experimental stream

Abstract Artificially hatched and reared juvenile European grayling, Thymallus thymallus (L.), from three natural populations were released into an experimental stream. Two of the populations were from lakes and the third population was from a river. Post-stocking dispersal was studied by recapturing fish in traps and by electric fishing after each of the stocking exercises. On all three occasions the first fish to be recaptured were caught in a trap about 400 m downstream from the release site, within 5–12 min of release. Proportions of stocked grayling juveniles recaptured by electric fishing in the fast-flowing section of the experimental stream (the rapids) were 9.9%, 46.7% and 16.6% after the first, second and third stocking, respectively. No significant population-specific difference was found between fish caught in the downstream trap and fish recaptured in the rapids. The uppermost part of the rapids (stretch 1) and the catch in the upstream trap was dominated by fish of riverine origin. Post-stocking migration was influenced by sex but not by body size. Females showed a greater propensity to stay in the rapids compared with males. Implications of the results for stocking grayling in natural streams and regulated rivers are discussed.     

Roles of discharge and temperature in recruitment of a cold‐water fish,the European grayling Thymallus thymallus,near its southern range limit

Recruitment of salmonids is a result of density‐dependent factors, specifically egg production in the previous year, and density‐independent environmental processes driven by discharge and temperature. With the plethora of knowledge on major drivers of Atlantic salmon Salmo salar and brown trout Salmo trutta recruitment, there is a requirement to explore less known species, such as European grayling Thymallus thymallus, whose postemergence time coincides with period of increasing temperature and low discharge. This study assessed drivers of grayling recruitment in a southern English chalk stream, a system vulnerable to discharge and temperature alterations under future climate change predictions. The analyses explored age 0+ grayling survival in relation to conspecific and heterospecific densities and discharge‐ and temperature‐derived factors. The final mixed‐effects model revealed a positive relationship between age 0+ grayling survival and incubation temperature anomaly and age 0+ trout abundance. Similarly, postincubation temperature anomaly had a positive effect on 0+ grayling survival, but only up to a threshold temperature of 13.5°C, beyond which it had a negative effect. In contrast, increasing number of days with low discharge postincubation negatively influenced age 0+ grayling survival, with no evidence of an effect of elevated discharges following spawning. Our results emphasise the importance of maintaining natural discharge regimes in salmonid rivers by tackling multiple stressors operating at the catchment scale, including land and water use to mitigate for predicted climate driven changes. In addition, further research on recruitment drivers in less stable, rain‐fed systems, is required.     

Effects of rearing technique and fish size on post-stocking feeding, growth and survival of European grayling, Thymallus thymallus (L.)

European grayling, Thymallus thymallus (L.), were reared by four different techniques: (A) in a natural pond without a supply of artificial diet; (B) in a net pen fed artificial dry diet; (C) in tanks fed artificial dry diet; (D) in tanks, and for 16 days before stocking, transferred to a diet based on mixed frozen natural food exclusively. Fish within groups C and D were also split into subgroups of large and small fish. Feeding, growth and survival of fish in the different groups were compared after stocking into three different stocking sites during the spring of 1994. The stocking sites consisted of a barren lake (Lake L), a second lake (Lake N) containing an allopatric perch, Perca fluviatilis L., population, and a barren man-made pond. In Lake L, higher recapture rates and mean weights of stomach contents were obtained in fish reared in the natural pond and in the net pen. There were no differences in total recapture rate, or mean condition factor between small and large fish within groups C and D. In Lake N only five (0.5%) stocked grayling from rearing groups A and B were recaptured during the experiment and more fish from categories C and D were recaptured. This was probably related to size-selective predation. In the pond, there were no differences in recapture rates or in mean weight of stomach contents between fish from any of the groups. In sample 1 (Lake N, 22 days after stocking), mean weight of stomach contents was higher in the larger fish within categories C and D but there was no difference in condition factor between the groups. In sample 1, mean condition factor of fish from the different groups varied markedly, being highest in Lake L followed by Lake N and the Pond. Fourteen out of 26 dead fish in the pond showed clinical symptoms of the bacterial disease ASA. Mortality was related to low initial condition factor at the time of stocking. No stocked grayling were recaptured from the two lakes in the last sample (June 1995).     

Putative virulence factors of atypical Aeromonas salmonicida isolated from Arctic charr,Salvelinus alpinus (L.), and European grayling,Thymallus thymallus (L.)

Atypical Aeromonas salmonicida (AAS) causes generalized lethal infections in farmed Arctic charr, Salvelinus alpinus (L.), and European grayling, Thymallus thymallus (L.), and is thus a serious threat for culture of these fish species. Virulence factors were studied among isolates of AAS from Arctic charr (n = 20), European grayling (n = 19) and other fish species (n = 20), of which 48 were of Finnish and 11 of Swedish origin. All isolates produced an A-layer. Extracellular products (ECP) of the AAS isolates did not produce detectable gelatinase and caseinase activity in test assays. Analysis of the same ECP preparations with substrate sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed weak proteolytic activity, indicating the different sensitivity of the detection methods used. The ECP from AAS isolates showed low cytotoxic activity against cultured cells. However, the ECP did not induce mortality in challenged Arctic charr. The results suggest that toxic components, like ECP, secreted by the bacterium may not be the major virulence factor in AAS-infection in Arctic charr and European grayling, and hence the pathogenesis also differs from the pathogenesis of AAS-infection in Atlantic salmon, Salmo salar L.     

Controlled reproduction in the wild European eel (Anguilla anguilla): two populations compared

This study aimed to describe the response variability of female silver eels in terms of gonad development and eggs production to a standardized gonadotropic treatment (Carp pituitary extracts—CPE), and to relate this variability to population characteristics. For this purpose, sexual maturation, ovulation, and fertilization were induced in two eel populations coming from different locations in Adriatic Sea (Comacchio—CM and Marano-Grado—MG lagoons), and after that, their reproductive capacity was valuated. External (Silver index—SI, Eye index—EI, Pectoral fin length index—PFLI, Condition factor—K) and hormonal (17β-estradiol—E2, testosterone—T) parameters were measured, and some subject/group were killed for histological and lipid analysis and age determination. Morphometric parameters showed the CM-Group to have highest values of Body weight (BW), Body length (BL), and K, while MG-Group presented highest PFLI and Gonadosomatic index (GSI) values. Regarding hormonal analysis, the CM-Group showed significantly higher T and E2 levels than the MG-Group, both groups showed considerably rapid increase at T5 (5th injection). A positive trend in gonadal development was found through histological evaluation; a more regular maturation was observed in the MG-Group, whereas the CM-Group presented an exponential oocytes development starting from T10 (10th week), which led to an anticipated spawning. Lipid content showed significant differences in T0 (start study), post-ovulation, and Control (30th week) between CM and MG eels. As to zootechnical performances, while MG eels released spontaneously into the water, the CMs were stripped in order to check ovulation. The MG eels were statistically the most productive with 40.1 ± 6.33 % BW of eggs released. Furthermore, CM females ovulated mainly between the 19th and 22nd week (77.8 % spawned eels) instead in the MG’s ovulation goes from the 24th to the 28th week (100 % spawned eels). As fertilization is of concern, in both groups fertilized eggs were obtained with no difference in larvae production. These results seem to indicate that bigger dimensions, higher K, and larger lipid content (Comacchio eels) could fasten gonadic maturation without positively influencing reproductive performance of animals, both in term of quantity and quality of produced eggs. Smaller females with a highest SI (Marano-Grado eels) presented a more regular gonadic development, leading the animals to spontaneous spawning.     

Post-stocking behaviour of hatchery-reared European grayling, Thymallus thymallus (L.), and brown trout, Salmo trutta L., in a semi-natural stream

Hatchery-reared, juvenile European grayling, Thymallus thymallus (L.), and brown trout, Salmo trutta L., were each stocked six times into an area of a semi-natural stream. The order in which the two species were released was switched after every second experimental stocking. Temporal and spatial post-stocking dispersal, effects of previously stocked species, feeding behaviour and the influences of sex and size were studied. During each 48-h experimental release period, some fish were recaptured in a trap situated 200 m downstream from the stocking site, and fish remaining in the stream after each experimental release were caught by electric fishing. Significantly more grayling than trout moved downstream and left the semi-natural stream. Proportions of stocked grayling recaptured in the trap within 2 h and from 2 to 48 h post stocking in the stream were 36.4% and 10.0%, respectively. Corresponding recapture rates for brown trout were both 1.5%. Most of the grayling and brown trout that did not leave the stream early were recaptured in deep, slow-moving water at low velocities in the release area. The presence of grayling at the time that the brown trout were stocked resulted in significantly fewer brown trout staying in the upper part of the stream. Within 48 h of their release, 33.3% and 22.8% of the grayling and brown trout, respectively, had eaten natural food items. The mean length of brown trout recaptured in the upper part of the stocking area was significantly lower than that of fish recaptured in the lower part and in the trap. Among brown trout, males showed a significantly greater propensity to eat and to stay in the upper part of the stream near the stocking site compared with females . Brown trout with natural food items in their stomachs had significantly lower mean length than trout without such items. No sex- or size-related differences were found in the spatial distribution or feeding activity of grayling.     

Genetic comparison of wild and cultivated European populations of the gilthead sea bream (Sparus aurata)

This study represents the first large-scale population genetic analysis of the marine fish gilthead sea bream (Sparus aurata), one of the most significant species in the South European aquaculture. Six wild and five cultivated sample sets covering the South Atlantic and Mediterranean European area have been screened for allozyme, microsatellite and mitochondrial DNA (mtDNA) variation. Microsatellites showed higher levels of polymorphism than allozymes. The low variability of mtDNA offered no basis for population differentiation.

The results reveal levels of variability for S. aurata above those from other sparids. Cultivated populations show a slight decrease of variability related to the wild ones, but not sufficient to document inbreeding depression effects, thus suggesting a fairly proper management. Wild populations reveal a slight degree of differentiation more pronounced with microsatellites than with allozymes, but not apparently associated with geographic or oceanographic factors. The cultivated populations seem to be highly divergent as a result of genetic drift caused by different factors pertaining to their respective histories. With both markers, the two cultivated Spanish sample sets are the most divergent. The high differentiation between cultivated and wild populations from the same area might indicate no evidence for significant genetic flow between them.

This study provides an insight into the population structure of S. aurata, although more questions have arisen that need to be solved. This can be achieved by further screening of small-scaled targeted sample sets in the studied area.     

投喂频率对凡纳滨对虾生长和水质的影响

研究了不同投喂频率(1,2,3,4和5次·d-1)对凡纳滨对虾生长、存活率和水质的影响。试验在室内2m3的圆形水泥池(d=1.6m)中进行,采用全封闭养殖模式,凡纳滨对虾(初始重量为0.24g)养殖密度为50尾·m-3,投喂人工配合饲料(粗蛋白含量为42.98%)。通过7周的试验,结果表明,投喂频率为1～4次·d-1时,随着投喂频率的增加,对虾增重率显著增加;投喂频率由4次·d-1增加到5次·d-1时,对虾增重率略有增加,但差异不显著。凡纳滨对虾的成活率在1次·d-1时最低,2次·d-1时最高,2～5次·d-1时随着投喂频率的增加而降低(投喂频率为1和2次·d-1之间、3和4次·d-1之间差异显著)。投喂频率为3次·d-1时饲料系数最低,蛋白质效率最高。随着投喂频率1～3次·d-1增加,饲料系数逐渐显著降低,蛋白质效率显著递增;而投喂频率从3～5次·d-1增加时,饲料系数增大,蛋白质效率逐渐下降(3和4次·d-1差异显著;4和5次·d-1之间差异不显著)。投喂频率对水环境中的pH值、H2S含量、COD、盐度等水质指标没有显著影响;在养殖后期水环境中投喂频率3～5次·d-1的NH 4和NO-2浓度较投喂频率1～2次·d-1时高。     

倒刺鲃的胚胎发育及水温对胚胎发育的影响

对倒刺鲃的胚胎发育进行连续跟踪观察,并探讨了水温对其胚胎发育的影响。结果表明:水温(25±0.5)℃时,孵化时间为53.43 h;胚胎发育过程中出现克氏囊。水温21～33℃条件下,倒刺鲃的胚胎均能孵化,水温(x)与孵化时间(y)成极显著的负相关关系:y=235.72 e-0.056 6x,P<0.01;水温与积温的关系为:y=7 519.4x-0.519 2,P<0.01。孵化率(y)与水温(x)相关关系式为:y=-1.665 5x2+88.621x-1 091.5,由曲线方程可以推算出胚胎发育的适宜水温范围为23.4～29.8℃,通过曲线求导数推算极值的方法,求得胚胎发育的最适水温为26.6℃。     

基于SSR和ISSR标记对江苏大竹蛏野生群体和人工增殖群体的联合分析

为探索人工增殖活动对本地大竹蛏(Solen grandis)种质资源带来的影响,采用SSR及ISSR分子标记手段对江苏大竹蛏野生群体与人工增殖群体的遗传多样性及遗传结构进行了联合分析。筛选的14对SSR引物扩增获得多态性位点150个(PPB=93.16%),Nei's基因多样性指数和平均Shannon信息指数分别为0.134 2和0.253 5;筛选的10条ISSR引物获得多态性位点85个(PPB=91.93%),Nei's基因多样性指数和平均Shannon信息指数分别为0.131 0和0.230 2。两种方法分析得到野生群体的遗传多样性参数均显著高于人工增殖群体,但两群体间并未产生明显的遗传分化。实验结果表明,江苏沿海大竹蛏群体的遗传多样性水平较高,人工增殖群体的遗传多样性虽有所下降,但并未导致该地区大竹蛏种质资源遗传结构的显著改变。因此人工增养殖活动仍然能够作为补充大竹蛏资源的主要手段,但应探索采用高效生态的增养殖模式,对大竹蛏的种质资源进行合理的开发与利用。     

Microsatellite genetic variation in wild and hatchery populations of the sea cucumber (Apostichopus japonicus Selenka) from northern China

Farming of the sea cucumber Apostichopus japonicus (Selenka) started 20 years ago and is still in rapid expansion in China. In order to assess the genetic status of both wild and cultivated stocks of this species, we used eight microsatellite markers to estimate the level of genetic diversity within five hatchery stocks and two wild populations of A. japonicus, and compared the degree of genetic differentiation between them. High levels of polymorphism were observed over all loci. The mean alleles and expected heterozygosities over the seven stocks were 10.4–12.3 and 0.735–0.783 respectively. The results of the microsatellite survey provide no evidence to show that hatchery practice of the sea cucumber in China to date has significantly affected the genetic variability of the cultured stocks. Significant differentiation was found between most pairs of the hatchery stocks and wild populations (F_st range: 0.008–0.036), and no obvious difference was detected between the wild populations (F_st=0.008). The information on the genetic variation and differentiation obtained in this study can be applied for future genetic monitoring of A. japonicus aquaculture stocks and will be useful for future genetic improvement by selective breeding, and for designing suitable management guidelines for these genetic materials.     

The effect of ozone on water quality and survival of turbot (Psetta maxima) maintained in a recirculating aquaculture system

Turbot, Psetta maxima, represent a valuable and growing subsector of global finfish aquaculture, although bacterial infections such as edwardsiellosis have adversely affected the industry in recent years. During an experiment designed to investigate the effect of direct ozonation on fish performance in RAS, a bacterial disease outbreak (Edwardsiella tarda) occurred, presenting an opportunity to record additional effects of experimental ozonation regimes on performance of turbot grown in RAS. This short note thus collates phenomenological information on survival, growth and water quality parameters recorded during a 91 day experiment with juvenile fish. Alongside antibiotic therapy, a high ozone treatment (360 mV) improved survival of stock compared to those in a non-ozonated control (200 mV) and significantly so compared to low ozone treatment (320 mV). Both experimental treatments reduced total heterotrophic and Vibrio sp. bacterial loading and nitrite concentration in culture water compared to the control. Experimental ozone treatment also suggested a trend for improved growth and feed intake. Although no confirmed link or mechanism between ozonation and reduced impacts of bacterial infection are proven in this study, the observations add further evidence to the body of work demonstrating beneficial effects of ozonation on water quality, survival and growth of farmed fish.     

文蛤养殖群体和野生群体遗传多样性的AFLP 分析

应用AFLP标记技术对辽宁和山东沿海文蛤（Meretrix meretrix）养殖群体和野生群体的遗传多样性进行分析。采用7对AFLP引物组合对5个群体（3个野生群体,2个养殖群体）150个个体进行扩增,共得到364个的位点。5个群体内的多态位点比例为84．3945％～87．6465％,总多态位点比例为99．6300％。群体的Nei’s基因多样性指数（H）为0．2301～0．2634;群体的Shannon多样性指数为0．3617～0．4248,群体间的遗传距离为0．0394～0．1609,群体内个体间的遗传距离为0．2592～0．5360。辽宁大洼野生群体和山东河口野生群体的多态位点比例、Shannon多样性指数和群体内遗传距离均高于辽宁盘山和辽宁庄河养殖群体,辽宁庄河野生群体的遗传多样性处于中等水平。用UPGMA方法构建的群体系统进化树显示,辽宁庄河野生群体单独成为一支,辽宁大洼野生群体与庄河养殖群体聚到一起,辽宁盘山养殖群体与山东野生群体聚到一起,但是用这5个群体的150个个体进行的聚类结果显示,所有个体基本是随机交叉聚类,不能形成明显的类群分支。分子变异分析（AMOVA）表明,文蛤群体94．04%的变异来源于群体内,群体间的变异仅占5．96％。以上结果表明,文蛤群体内遗传多样性非常丰富,群体间相似性较大,且存在较强的基因交流。[中国水产科学,2008,15（2）：215-221]     

An investigation on the gamete quality of Black Sea trout (Salmo trutta labrax) broodstock fed with mealworm (Tenebrio molitor)

In this study, the effect of combined feeding Black Sea trout (Salmo trutta labrax) broodstock with commercial feed and mealworm larvae (Tenebrio molitor) for 50 days before the breeding period in order to evaluate the gamete quality. While the control group was fed with commercial feed, mealworm larvae were given as an additional protein source 2 days a week and 3 days a week to the experimental groups formed from female and male individuals. In addition to the growth parameters, the number of eggs (number/individual) and the egg diameter (mm), sperm volume (ml), density of spermatozoa (×10⁹ cell/ml), total motility (%), progressive motility (%) and average curvilinear velocity (VCL, µm/sec) values were determined. At the end of the study, the highest live weight gain was found as 46.2 g in the control group of male. The egg diameters were 4.3 ± 1.8 mm and 4.5 ± 1.4 mm in the worm treatment groups fed twice and three times with mealworm respectively. As a result, it was determined that feeding fish with mealworm larvae as an addition to the commercial diet in female individuals did not affect the amount of eggs (p > .05), but the egg diameters were significantly smaller in the control group than the mealworm groups (p < .05). In spermatological characteristics, only the amount of sperm was different between the groups (p < .05); all other parameters were found to be similar to each other with no statistical differences (p > .05).     

Differential distribution and response to experimental sexual maturation of two forms of brain gonadotropin-releasing hormone (GnRH) in the European eel,Anguilla anguilla

Using specific radioimmunoassays for the two GnRH molecular forms present in the European eel, Anguilla anguilla, (mGnRH and cGnRH II), we compared their distributions in the pituitary and different parts of the brain of female silver eels, as well as the modifications of their levels in experimentally matured female eels (treated with carp pituitary extract). In control eels, mGnRH levels were higher than cGnRH II levels in the pituitary, olfactory lobes and telencephalon, di- and mesencephalon, while the opposite was found in the posterior part of the brain (met- and myelencephalon). Experimental sexual maturation of the gonads significantly increased mGnRH levels in the pituitary and anterior parts of the brain; such a positive effect was not observed on the low cGnRH II levels, which were, in contrast, reduced. These data indicate that the positive feedback of gonadal hormones on GnRH, that we previously demonstrated, would specifically affect the mGnRH form. The differential distribution and control of mGnRH and cGnRH II suggest that these two forms have different physiological roles in the eel. The large increase in mGnRH during sexual maturation suggests the prime implication of this form in the neuroendocrine control of reproduction.     

Genetic diversity of cultured and wild populations of the giant freshwater prawn Macrobrachium rosenbergii (de Man, 1879) based on microsatellite analysis

Freshwater prawn (Macrobrachium rosenbergii) culture in the Western Hemisphere is primarily, if not entirely, derived from 36 individual prawns originally introduced to Hawaii from Malaysia in 1965 and 1966. Little information is available regarding genetic variation within and among cultured prawn stocks worldwide. The goal of the current study was to characterize genetic diversity in various prawn populations with emphasis on those cultured in North America. Five microsatellite loci were screened to estimate genetic diversity in two wild (Myanmar and India‐wild) and seven cultured (Hawaii‐1, Hawaii‐2, India‐cultured, Israel, Kentucky, Mississippi and Texas) populations. Average allelic richness ranged from 3.96 (Israel) to 20.45 (Myanmar). Average expected heterozygosity ranged from 0.580 (Israel) to 0.935 (Myanmar). Many of the cultured populations exhibited reduced genetic diversity when compared with the Myanmar and the India‐cultured populations. Significant deficiency in heterozygotes was detected in the India‐cultured, Mississippi and Kentucky populations (overall F_is estimated of 0.053, 0.067 and 0.108 respectively) reflecting moderate levels of inbreeding. Overall estimate of fixation index (F_st = 0.1569) revealed moderately high levels of differentiation among the populations. Outcome of this study provide a baseline assessment of genetic diversity in some available strains that will be useful for the development of breeding programmes.