Monitoring of the genetic variability of the hatchery and recaptured fish in the stock enhancement program of the rare species barfin flounder <Emphasis Type="Italic">Verasper moseri</Emphasis>

ABSTRACT:   In relation to the stock enhancement program for barfin flounder, hatchery juveniles produced in 2001 were genotyped using microsatellite DNA markers (msDNA) and then released to natural waters. Subsequently, recaptured individuals, designated as 'tentative recaptured', were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, pairwise F _ST and genic differentiation tests were used to estimate the genetic divergence between the wild samples, the hatchery broodstock, and the tentative recaptured samples. Analysis showed significant differentiation among these three groups. Pedigree determination by msDNA was used to establish the origin of the tentative recaptured individuals, in order to elucidate whether they were hatchery produced, 'real recaptured', or wild specimens. Wild individuals were not found. The effective population size of the real recaptured stock was very low ( N _e = 16.6). Equal family survivability was observed between the released and real recaptured stocks, indicating that the genetic variability of the released stock was maintained in the natural environment. Future broodstock management, breeding designs, and family contribution equalization of the offspring to be released will be required in this rare species to avoid unintended genetic differentiation between the wild population and the hatchery broodstock.     

Genetic assessment of milkfish (Chanos chanos Forsskal) stocks based on novel short tandem repeats for marker‐aided broodstock management

Milkfish hatchery broodstock are either from on‐grown wild‐caught or hatchery‐produced fry/juveniles. To determine if a marker‐assisted management scheme can be formulated for improved milkfish hatchery production, milkfish stocks were genetically characterized using nine novel short tandem repeats or microsatellites. Eight wild‐bred Philippine stocks (CLA, CUR, CAM, SIH, SBH‐I1, HH, PAL and ZH‐P₀), four hatchery‐bred stocks (SBH‐I2, SBH‐D, BoH and ZH‐F₁), two farm stocks of known mixed lineages (SPH and BDH) and one Indonesian hatchery‐bred stock (WJH) were assessed. WJH was included since milkfish fingerlings from Indonesia reared in Philippine farms could be developed into future broodstock. Mean allelic richness (Ar) was highest in wild‐bred stocks (9.5) and lowest in hatchery‐bred spawners (9.1). Mean expected heterozygosities (He) were relatively similar in all stocks with wild‐bred stocks slightly higher (0.67) than the others. An analysis of molecular variance indicated significant yet low genetic differentiation among stocks (F_ST = 0.013; p = .000) where variation (98.6%) was explained by intra‐stock differences. In some of the domesticated stocks, reductions in mean allelic richness were observed in first generation hatchery broodstock (e.g. ZH‐F₁; Ar = 8.3), compared with their founder stock (e.g. ZH‐P₀; Ar = 9.4). The Indonesian stock was similar to local wild‐bred stocks based on genetic variability indices; thus, it might be likely that the local stocks’ fitness traits could be comparable with the imported milkfish stock which has been perceived to be better. The quality of locally available farmed milkfish and prospects of formulating a broodstock management scheme for the production of good quality milkfish seedstock are herewith discussed.     

Genetic variation in farmed populations of the gilthead sea bream Sparus aurata in Greece using microsatellite DNA markers

Genetic variation in seven reared stocks of gilthead sea bream Sparus aurata, originating from Greek commercial farms, was assessed using five polymorphic microsatellite markers and was compared with that of two natural populations from the Ionian and the Adriatic Seas. The total number of alleles per marker ranged from 11 to 19 alleles, and hatchery samples showed the same levels of observed heterozygosity with samples from the wild but substantially smaller allelic diversity and expected heterozygosity. The global genetic differentiation for the cultivated samples was significant as indicated by F_st analysis, which might indicate random genetic drift and inbreeding events operating in the hatcheries. On the contrary, no significant difference was found between the two wild populations. Population pairwise tests between farmed and wild stocks were also significant, with the exception of one hatchery sample, the Central Greece 1, which was not significantly different from the two wild samples perhaps due to its recent use in aquaculture from wild‐caught animals. The UPGMA tree topology grouped the wild samples together with the Central Greece 1 stock, and showed a clear division between wild and farmed sample sets for the six remaining hatchery samples. Knowledge of the genetic variation in S. aurata cultured populations compared with that in the wild ones is essential for setting up appropriate guidelines for the proper monitoring and management of the stocks either under traditional practices or for the implementation of selective breeding programmes.     

Genetic diversity and relatedness estimates for captive barramundi (Lates calcarifer,Bloch) broodstock informs efforts to form a base population for selective breeding

Aquaculture of barramundi or Asian seabass (Lates calcarifer) is growing in both Australia and Southeast Asia and there is substantial interest to improve production efficiency through selective breeding. The establishment of a large and genetically diverse base population is a prerequisite for a sustainable and long‐term productive breeding program. Before selective breeding programs can begin for Australian barramundi it is important to assess the overall genetic diversity of current captive broodstock populations. To address this question, 407 captive barramundi broodstock from eight separate Australian broodstock populations were genotyped using 16 polymorphic microsatellite DNA markers. A Bayesian STRUCTURE analysis indicated that captive Australian broodstock are broadly divided into two genetic stocks. Multivariate analysis between broodstock individuals and pairwise F_ST between broodstock populations also supported the existence of two stocks. Comparisons with data obtained from natural stocks suggested that hatchery individuals were either sourced from the two stocks or represented an admixture between them. Genetic diversity was low within each broodstock population (allelic richness ranged from 2.67 to 3.42 and heterozygosity ranged from 0.453 to 0.537) and relatedness estimates within hatcheries were generally low (average r was equal to 0.141). We recommend sourcing captive individuals according to high levels of neutral genetic diversity and low levels of relatedness for the establishment of a base population. We also make recommendations about including genetically diverse wild individuals.     

Loss of genetic diversity due to hatchery culture practices in barramundi (Lates calcarifer)

Many aquaculture hatchery practices are detrimental to the long-term viability of restocking and selective improvement programs. Small effective broodstock population sizes, differential broodstock contribution, differential larval/juvenile survival during metamorphosis and size-based grading, all have the potential to drastically reduce the level of genetic variation remaining in hatchery populations. Monitoring levels of genetic variation and maintaining detailed pedigrees on progeny is the key to circumventing these problems. In this study we used microsatellites, coupled with DNA parentage analyses, to track the loss of genetic diversity in two independent commercial barramundi (Lates calcarifer) hatcheries over three mass spawning events, where up to two females and seven males had the opportunity to participate. Initial broodstock contributions were observed to be highly skewed, with significant differences observed in both the level of contribution by females to each mass spawning, as well as in the number of males participating and subsequently contributing to the genetic composition of cohorts. Effective population sizes were around half that of census sizes. We then examined whether differential family survival through metamorphosis (27 days post-hatch) and/or first size grading further influenced the retention of genetic diversity levels initially sampled during spawning. Parentage analyses indicated that some families that had been initially represented in cohorts had been lost, or that the contribution by particular broodstock had changed. In one cohort, as many as 55% of progeny were found to be sired by a single male individual. Size grading was also found to potentially impact on genetic diversity, with data suggesting that family representation in each of the grades was non-uniform and that some families were on average faster or slower growing than others. These results illustrate that hatchery management practices have the potential to significantly impact on the retention of genetic diversity in this species.     

Effect of introducing wild paternity on stock performance of hatchery-reared ayu

The domestication of ayu Plecoglossus altivelis, in which breeding has been carried out for multiple generations without the introduction of exotic broodstock (purebred-styled cultivation), causes rapid loss of genetic variability, and is problematic for resource management programs. The introduction of wild paternity to enhance genetic variability was examined for its effects on the performance of the newly hybridized stock using the Gunma population (hatchery strain), which has been inbred for 29 generations since its origin. Principal component analysis showed that hybridization could modify the stock performance, moderately affecting behavior and morphometrics. Reinforcement of aggressiveness as well as transformation to a long-nosed shape in the hybrid population contribute drevised performance that was acceptable to sports fishers as an artificial stock. To reduce genetic stress for native ayu populations, the introduction of wild paternity is proposed through sperm from wild caught specimens to hybridize with eggs from the pure-bred hatchery population.     

Maintenance of genetic variation in hatchery stocks of Atlantic salmon, Salmo salar L.: experiences from the River Bush, Northern Ireland

Abstract. An examination of biochemical-genetic variation at seven polymorphic loci was carried out among five year classes of wild Atlantic salmon, Salar L., in the R. Bush and in a hatchery strain derived from the wild population. Within some of the year classes, gene frequencies at several loci differed significantly between wild and artificially reared salmon. Highly significant temporal variation in gene frequencies was detected among successive year classes of the hatchery strain, while this was less significant among the wild salmon. Samples of wild salmon taken as 0+ and 1 + parr in the river showed no significant temporal variability in allelic frequencies. Heterozygosity levels among the wild and hatchery-reared salmon were comparable, averaging 0·185 and 0·176 respectively. The genetic variability of the artificially reared salmon is discussed in relation to numbers of broodstock and breeding regime used at the hatchery.     

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.     

Genetic variation of hatchery and wild stocks of the pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis> (Dunker, 1872), assessed by mitochondrial DNA analysis

In order to provide baseline information for the genetic resources, genetic variation in wild and cultured Pinctada fucata martensii from southern Korea and Japan was studied using nucleotide sequence analysis of 379 base pairs (bp) in the mitochondrial cytochrome oxidase subunit I gene (COI). The study included three hatchery stocks from Korea (Tongyeong) and Japan (Mie and Tsushima) and one wild stock from Korea (Geoje). A total of 3 haplotypes were identified in hatchery stocks of 78 individuals, of which 63 individuals shared 1 haplotype. Overall, nucleotide diversity (π) was low, ranging from 0.000 to 0.002, and haplotype diversity (h) ranged from 0.000 to 0.541. Considerably low haplotype and nucleotide diversities in hatchery stock indicated that low effective population size and consecutive selective breeding of P. fucata martensii could be responsible for the reduction in genetic variation. The wild stock exhibited low haplotype diversity (0.507 ± 0.039) with two shared haplotypes. The results of the present study with first record of wild pearl oyster in Korea support the possibility that the transplanted pearl oyster for overwintering experiments could have survived in winter. In order to enhance and/or maintain genetic diversity in the hatchery stock, further research should be directed toward genetic monitoring and evaluation of the hatchery and wild pearl oysters.     

Japanese chum salmon stock enhancement: current perspective and future challenges

This study reviews the present status of the Japanese chum salmon Oncorhynchus keta stock enhancement program and considers the ecological sustainability of wild populations while providing fishery production, exemplified by the hatchery-based Kitami region set net fishery. The return rate and the number of returns have been historically high in the Sea of Okhotsk, but have decreased in other regions since 2005. Natural spawning of chum salmon occurred in at least 160 rivers in Hokkaido. The genetic diversity of Japanese chum salmon was similar to or higher than that of other Pacific Rim populations. Numbers of alleles were high at microsatellite loci, but the loss of rare haplotypes was observed in all populations. The estimated N _e /N ratio for the Kitami region was >0.15 % including hatchery and wild fish under the present high fishing pressure. Four regional populations were inferred in Hokkaido, however, genetic differentiation was weak and some river-populations were nested. Substantial changes in run timing were observed, but it has recovered gradually owing to the recent practice of escapement. Our analyses highlight the importance of juvenile quality and the vital roles of escapements in enhanced and non-enhanced rivers. New research is needed to minimize the genetic risks associated with hatchery programs.     

基于SSR标记的长江下游原良种场鳙亲本和后备亲本种质资源现状分析

分析长江下游原良种场鳙(Aristichthys nobilis)亲本和后备亲本的遗传多样性和遗传结构,评估鳙育种群体的种质资源现状,可为鳙的种质资源管理和活体资源库建设提供科学的参考依据。本研究利用已发表的10对SSR荧光引物结合毛细管电泳技术,对7个亲本群体和1个后备亲本群体共638份DNA进行基因分型。主要应用GenAlEx 6.501、Cervus 3.0和Structure 2.3.4等软件进行遗传多样性参数计算和群体遗传结构分析。结果表明,8个群体总体遗传多样性水平较高,但存在一定程度的近亲繁殖风险。其平均等位基因数(N_a)为14.83±1.45,观测杂合度(H_o)和期望杂合度(H_e)分别为0.76±0.05、0.82±0.02,Shannon's信息指数(I)为2.08±0.09,近交系数(F)为0.08±0.05。各亲本群体之间遗传多样性水平存在差异,后备亲本遗传多样性水平最高。鳙的贝叶斯群体遗传结构分析图(Structure)与主坐标分析(PCoA)结果具有一致性,将所有样本划分为两个类群,大部分亲本群体之间并无明显的遗传分化,而后备亲本和亲本之间则表现明显的遗传分化。分子方差分析(AMOVA)显示,11%的遗传变异来自群体间,群体间的遗传分化处于中等水平,而89%的遗传变异来自群体内。本研究探究了鳙亲本和后备亲本的遗传差异性,对野生原种引进和保护提供了建议,以期为构建健康负责的增殖放流种苗繁育体系提供理论支持和数据参考。     

Use of Microsatellite DNA Profiling to Identify Japanese Flounder,Paralichthys olivaceus of Hatchery Origin

Japanese flounder, Paralichthys olivaceus, is an economically important marine fish species in Asia. A suite of 18 microsatellite markers chosen from published genetic linkage maps was used to carry out parentage assignments of 188 hatchery‐reared juveniles from a small number of breeders. The probabilities of exclusion for the 18 microsatellite markers were 0.604–0.913, and the effectiveness of combined probability of exclusion reached 100% when using the eight microsatellite markers with higher Excl 1 probabilities. The cultured and wild stocks (WSs) were differentiated in a release‐recapture population based on these markers. Of the 321 recaptured offspring, 28.34% were assigned to their parental pairs in our broodstock, whereas the remaining offspring could not be traced back to a possible sire or dam. Significant reduction in genetic diversity of the cultured stock (CS) had not been found compared with that of the WS. The results suggest that CSs released into the wild will not adversely affect the genetic structure of natural populations. Our results demonstrate that these markers provide an efficient tool for parentage assignments and genetic analysis of Japanese flounder.     

Loss of genetic variability in the captive stocks of tambaqui,Colossoma macropomum (Cuvier, 1818), at breeding centres in Brazil,and their divergence from wild populations

The loss of variability in farmed populations and the risks of interactions with wild populations support the need for the genetic monitoring of species farmed throughout the world. In Brazil, the tambaqui is the most widely farmed native fish species. Despite this, there are no data on the pedigree of the farmed stocks, and the potential for interactions with wild populations in the Amazon basin has raised concerns with regard to the genetic variability of these stocks. The present study analysed sequences of the mitochondrial Control Region and 12 microsatellites to characterize the genetic variability of seven historically important commercial tambaqui breeding centres located in four different regions of Brazil, and compared these sequences with those obtained from individuals collected from a wild population. High levels of genetic diversity were found in the wild population, whereas genetic diversity was reduced in both markers in most captive populations, except for the broodstock located near the Amazon River. High F_ST and D_EST indices were recorded between the wild population and most of the captive stocks analysed. The drastic reduction in genetic diversity found in most captive stocks and the difference between these stocks and the wild population may have been the result of the small size of the founding populations and the absence of breeding management. The renewal of the broodstocks and the application of breeding management techniques are recommended. In the Amazon region, in addition, the use of broodstocks that are genetically very different from local wild populations should be avoided.     

Influence of Hatchery Protocols on Mitochondrial DNA Variation in Japanese Flounder Juveniles

The Japanese flounder Paralichthys olivaceus is a commercially important fish that is stocked extensively from hatchery rearing programs in Japan. To examine the genetic variability of hatchery-raised juveniles of the Japanese flounder that are used for stocking into natural waters, we analyzed a portion of the mitochondrial genome. The mtDNA region extending from the 3'half of the cytochrome b gene to the central domain of the control region was PCR amplified and analyzed using 11 restriction endonucleases. We identified 34 polymorphic cleavage sites out of a total of 61 sites, which resulted in 67 different haplotypes in a total of 265 offspring, examined from eight hatchery stocks. Haplotype diversity of offspring at each of the eight hatcheries ranged from 0.49 ± 0.09 (SE) to 0.94 ± 0.03 (SE). Also, we observed 40 polymorphic sites out of a total of 59 sites, which resulted in a total of 50 haplotypes in 60 wild flounder. Haplotype diversity of the wild population was 0.98 ± 0.01 (SE). The use of subcultured fishes as broodstock appears to be one of the most important causes of reduced genetic diversity in hatchery-raised flounder juvenile. Our results suggest that the use of wild fish for broodstock is an effective way to maintain genetic variability in Japanese flounder offspring.     

Tench (Tinca tinca) broodstock management in breeding station under conditions of pond culture: a review

A basic assumption for carrying out the breeding work is to have well-organised broodstock management. Taking into account the number of populations bred and the limited number of ponds available, it is impossible to avoid rearing several juvenile or adult breeds in mixed stocks. Fish of individual breeds are group-marked with regular renewal of the freeze-branded mark and broodstock adults should be marked individually. To minimise inbreeding or losses in genetic variation, at least 120 fish per strain are reared and when the strain is restored, factorial crosses of at least 15 females and 25 males should be applied. All activities are individually registered in the “Evidence 2003” data-recording software and data can be sorted by the origin of the fish population, of the strain or of individual fish; reproductive and performance parameters are recorded individually also. Further breeding operations with tench comprise the selection after over-wintering and rearing of fish before, during and after the reproductive season.     

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.     

Straying of Atlantic salmon, Salmo salar, from delayed and coastal releases in the Baltic Sea, with special focus on the Swedish west coast

Abstract  Atlantic salmon, Salmo salar L., reared from two Baltic strains were released around the islands Bornholm and Møn in the Baltic Sea between 1995 and 1999. A total 600 000 reared salmon were released from net pens using the delayed release technique, keeping the salmon in net pens for approximately 3 months after smolting, and 208 000 were released directly from the hatchery. Of these, 15 958 were tagged with Carlin tags. Additionally, 65 300 coded wire tagged salmon were released as delayed release salmon close to Bornholm in 2000. Recaptures from the five years of Carlin tagged releases varied between 2.8% and 21.2% (average 13.1%). Most recaptures were from within the Baltic Sea (average 98%), but some were recaptured outside the Baltic Sea, either in the sea (1%) or in fresh water (1%). Recaptures outside the Baltic Sea and in fresh water were higher for releases at Møn in the western part of the Baltic, than releases at Bornholm. Straying rates from the releases into six rivers on the Swedish west coast were estimated using information from capture in traps and sport and broodstock fisheries. The proportion of straying salmon in rivers on the Swedish west coast was about 3.8% of the salmon run, but with large variations between rivers. Releases were discontinued because of possible deleterious effect on the local wild salmon populations.     

Genetic diversity of wild mekong giant catfish Pangasianodon gigas collected from Thailand and Cambodia

The Mekong giant catfish Pangasianodon gigas is endemic to the Mekong River and is a critically endangered species. The genotypes of the microsatellite DNA (msDNA) and mitochondrial DNA (mtDNA) markers (right domain of the control region) were detected to evaluate the present status of genetic divergence of this species from the Mekong River in Thailand and Cambodia. The observed and expected heterozygosity values of Mekong giant catfish in Thailand and Cambodia were relatively low in comparison with those of other nonendangered freshwater fish species. These two populations from Thailand and Cambodia showed similar levels of genetic diversity, as evaluated by the 384 nucleotides of the mtDNA control region with 13 haplotypes. The pairwise F _ST value between the two populations based on the genotype frequencies of msDNA and mtDNA markers suggested a close genetic relationship between the populations in Thailand and Cambodia. The results of this study support the conclusion that the Mekong giant catfish is critically endangered. Care should be taken to sustain the genetic diversity of this species, as the level of genetic variability has already decreased in the wild population. This species is a target species for an ongoing stock enhancement program in the Mekong River in Thailand. It is proposed to apply these markers for proper broodstock management, such as for minimal kinship selective breeding in the hatchery.     

Reproductive strategies in small populations: using Atlantic salmon as a case study

Abstract –  Wild salmonid populations with only a few breeding adults may not exhibit a significant reduction in genetic variability compared with larger populations. Such an observation suggests that effective population sizes are larger than population size estimates based on direct adult counts and/or the mating strategy maximises outbreeding, contributing to increased heterozygosity. In the case of wild Atlantic salmon Salmo salar populations, stratification by age classes and sexes on the spawning grounds avoids inbreeding and increases genetic variability. We studied the breeding composition of four Spanish salmon populations. Over a 7-year period we concluded that the probability of within-cohort mating is very low: females generally reproduce after two sea-winters whereas males reproduce mostly as one sea-winter ( grilse ) and/or mature parr. Considering different levels of contribution of mature parr to spawning derived from field surveys, we developed a simple model for estimating effective population sizes and found that they doubled with 65% parr contribution expected for rivers at this latitude (43°N), and ranged from 100–800 individuals. The effect of between-cohort mating was modelled considering different ranges of differences in allele frequencies between cohorts and resulted in 28–50% increases in heterozygosity when considering a 65% parr contribution. The complex mating strategy of Atlantic salmon contributes to explain the high levels of genetic variability found for small populations of this species. This model can probably be extended to other animal species with mating strategies involving different cohorts.     

Fecundity trends of Chinook salmon in the Pacific Northwest

Fecundity is an important demographic parameter that contributes to the productivity of anadromous fish stock dynamics. Yet, studies on fecundity patterns in Pacific salmon (Oncorhynchus spp.) often only include a few years of data, limiting our ability to understand spatio-temporal trends. Here, we used data on 43 hatchery Chinook salmon (O. tshawytscha, Salmonidae) populations in Washington State to evaluate whether average fecundity changed over the past three decades. We then used data from a subset of stocks (18) to evaluate the relationship between fecundity and body length. Our results revealed significant changes in fecundity across the 25-year study period with most stocks showing declines in fecundity over the past decade. Results further showed that Chinook salmon have decreased in length over this same period and that annual variation in mean length explains a majority (62%) of annual variation in mean fecundity. Specifically, we estimated that a 1-mm reduction in length results in 7.8 fewer eggs (95% CI = 6.6–8.9). Given that the majority of Pacific Northwest Chinook salmon in the environment and harvested in fisheries originate from hatchery releases and that nearby hatchery and wild populations generally have similar ocean distributions, these results likely reflect patterns for many populations not included. Combined, our results highlight the need to consider changes in body size and egg production when assessing the dynamics of anadromous fish populations and designing management or conservation plans, particularly for depressed populations.