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 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.     

Identification of differential broodstock contribution affecting genetic variability in hatchery stocks of Atlantic salmon (Salmo salar)

Supportive breeding of Atlantic salmon (Salmo salar) is commonly employed to maintain numbers of fish where the species has become locally endangered. Increasingly, one of the main aims of population management is the preservation of natural genetic diversity. If the stocks employed in supportive breeding exhibit reduced variation they can alter the natural pattern of genetic variation observed in wild populations. In northern Spain, wild adult salmon are caught every year from local rivers and artificially crossed in order to create supportive stocks. The offspring are hatchery reared until the juvenile stage, then released into the same river where their parents were caught. In the current study, our findings demonstrate that although adult broodstock exhibit a pattern of variation similar to the wild populations, variability at microsatellite loci was drastically reduced in the juveniles released into one of three rivers analyzed. The contribution of broodstock to this juvenile stock was examined by pedigree analysis. A restricted number of females contributing to the hatchery stock was identified as the main cause of loss in genetic variation, possibly due to overmaturity of some multi-sea-winter females. We suggest that better monitoring and control of parental contribution will help in solving the problem of loss of genetic diversity in hatchery populations.     

基于线粒体控制区序列的三疣梭子蟹增殖放流亲蟹遗传多样性研究

为研究山东沿海三疣梭子蟹增殖放流亲蟹群体的遗传多样性状况,实验采用536 bp的线粒体DNA控制区片段作为分子标记,对4个亲蟹群体的遗传多样性和遗传结构进行了分析。结果显示,301个三疣梭子蟹个体共检测到155个单倍型,4个群体的单倍型多样度为0.972 3~0.993 0,核苷酸多样度为0.021 2~0.023 6,表现出丰富的遗传多样性。AMOVA分析和Fst分析结果均显示,4个三疣梭子蟹亲蟹群体间遗传分化微弱,未形成明显的遗传结构,NJ系统树中未出现与各群体相对应的的谱系分支。研究表明,4个增殖放流亲蟹群体的遗传多样性丰富,且其遗传结构与放流海域的野生群体间没有明显分化,种质资源质量较好。此外,群体历史动态分析显示,渤海南部和黄海北部的三疣梭子蟹历史上曾经历过群体扩张事件。     

Erosion of the native genetic resources of brown trout in Spain

Abstract– We analyzed the introduction of hatchery-reared trout in the Riutort Creek, a small stream in the eastern Spanish Pyrennees. We used gene correlation matrices between individuals to analyze the fish coancestry in the Riutort Creek samples and in the hatchery stock. Hatchery fish disturbed the single ancestry in the native population of the creek, and were clearly detected with principal coordinate analysis of the gene correlation matrix. The amount of introgression produced by successful introductions was estimated from the principal coordinate analysis projections of the matrix of F_ST values between the putative native Riutort Creek population, the hatchery stock and the introgressed population. In only two years the amount of introgression rose to 10%, indicating that 5% of the native ancestry is lost each year as a result of the stocking program. Based on these results, we review the present understandings on the genetic impact of hatchery fish on indigenous Spanish brown trout populations. The stocking of these populations involves a non-native broodstock widespread through the Spanish hatcheries, but successful stockings appear to be limited to wild populations subjected to occasional releases in protected or unfished areas. Surprisingly, extensive stocking in fished areas result in a more limited genetic impact on the recipient native population.     

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.     

放流三疣梭子蟹月间回捕率及繁殖贡献率研究

2015年6月,将约500万只人工繁育的三疣梭子蟹C2期稚蟹放流到辽宁盘锦沿岸海域.当年7月28日—10月3日在盘锦沿岸以地笼网回捕1671只三疣梭子蟹样本,剪其螯足,剥离肌肉,提取DNA,利用微卫星DNA分子标记进行亲子鉴定,开展个体识别.试验结果显示,120只为当年放流蟹,放流后苗种短期月间放流蟹占当月回捕蟹群体的...     

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.     

基于微卫星标记对长江江苏段鳙增殖放流效果评估

为评估长江江苏段鳙(Aristichthys nobilis)增殖放流效果,本研究基于微卫星标记的亲子鉴定技术,选用 10对扩增效果稳定的微卫星荧光标记引物,结合毛细管电泳对江苏省内良种场 563 尾鳙亲鱼和长江江苏段 687 尾回捕鳙进行基因分型,并对位点多态性及亲子鉴定结果进行了分析。结果表明,各位点等位基因介于 19~55 之间,其观测杂合度为 0.530~0.909(平均值为 0.748),期望杂合度为 0.818~0.929 (平均值为 0.882),多态性信息含量为0.797~0.924 (平均值为 0.871);Cervus3.0 模拟结果显示,当双亲未知且置信度为 95%时, 10 个微卫星位点的累积非亲权排除率为 99.996%。亲子鉴定结果发现,所有回捕鳙中有 42 尾与亲本之间存在亲子关系,被确认来源于鳙增殖放流群体,并由此推算此次评估亲本的放流子代对长江江苏段野生群体的贡献率为 6.11%。结果显示该研究进行的鳙亲子鉴定技术可为长江鳙增殖放流效果评估工作提供可靠的数据支撑,为长江渔业资源管理提供参考资料。     

象山港黄姑鱼增殖放流效果评估及增殖群体利用方式优化

以回捕渔获重量和对繁殖群体的补充能力为评价指标,借助标志放流-回捕实验,利用模型模拟分析方法,定量评估浙江象山港黄姑鱼(Nibea albiflora)的增殖放流效果;同时,结合其增殖目标定位,探索构建生态高效的增殖群体利用方式。研究结果表明,象山港黄姑鱼增殖群体的捕捞死亡系数为1.31,在该捕捞强度下,11055尾增殖放流鱼苗可产生737 kg回捕渔获收益,同时还向增殖水域补充了554尾初次性成熟个体,增殖放流活动在一定程度上起到了修复象山港黄姑鱼资源、促进渔民增产增收的效果。结果表明,捕捞强度过大是制约象山港黄姑鱼增殖放流功效发挥的重要因素,捕捞死亡系数应降至0.46,同批次黄姑鱼增殖放流所能提供的回捕渔获重量和性成熟个体尾数可分别较现行捕捞强度提升41.49%和326.90%。综上述,象山港黄姑鱼增殖放流的生态和经济功效较为显著,合理降低对增殖群体的捕捞强度是进一步提升其增殖放流效果的必要措施。     

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.     

Increase of inbreeding by stocking on wild population assessed by using in dividual-based life history model

ABSTRACT:   The genetic impact of stocking on natural populations was assessed by using individual-based life history models. Models were constructed that included density-dependent mortality at the early life stage, natural mortality, fishing mortality, and the number of released fish. These variables were varied by using random numbers. The focus was on the genetic impact of the number of released fish, the number of parent fish producing seedlings that were released, the sex ratio of the parent fish, fishing mortality, and the methods to select the parents of the seedlings. When brood stock size in the hatchery was five males and five females, the coefficient of inbreeding was about 27-fold as high as that in the case of 50 males and 50 females at the 50th generation. The coefficient increased about fivefold at the 50th generation, when the sex ratio of the parents changed from 50 males and 50 females to 10 males and 90 females. When parents were taken from wild populations at random, extreme fishing mortality reduced genetic diversity.     

Genetic drift in a hatchery and the maintenance of genetic diversity in hatchery-wild systems

ABSTRACT:   Loss of within-population genetic diversity by genetic drift is related to stocking. In order to reduce it, selective use of wild-born individuals for hatchery broodstock is proposed. Its effectiveness is evaluated by the analytical method and numerical simulation. Results indicated that it is effective, especially when the ratio of released hatchery-produced individuals is high in the natural population. Loss of within-population genetic diversity by a few other strategies of broodstock preparation is also analyzed by numerical simulation and the effectiveness of these is also discussed.     

Population genetic structure and variability of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> in the stocking area along the Pacific coast of northern Japan

ABSTRACT:   The genetic diversity of wild and hatchery-released Pacific herring Clupea pallasii collected from three brackish lakes and two bays in Honshu and Hokkaido, Japan was examined with five microsatellite loci. All loci showed high genetic variability with expected heterozygosities ranging from 0.815 to 0.945. Significant differences in genotypic and allelic distributions were detected among all locations except for between the two bays in Honshu Island. Pairwise population analysis based on the F _ST values showed close genetic relationships among the locations in Hokkaido Island, and the hierarchical analyses of molecular variance showed significant genetic difference between the two islands. Those results suggest the existence of subpopulations due to natal homing. In addition, stocked fish showed as much genetic diversity as the wild fish. The pairwise population analyses also showed close relationships between the hatchery fish and the wild fish in respective stocking areas, showing that no effects of stocking programs on genetic diversity of wild populations were detected.     

Impacts of hatchery release on genetic structure of rock carp <Emphasis Type="Italic">Procypris rabaudi</Emphasis> in the upper Yangtze River,China

Rock carp Procypris rabaudi is a vulnerable endemic fish in the upper Yangtze River. Hatchery release has been carried out as a major stock enhancement strategy for this species. Ten microsatellite loci were chosen to compare genetic variation between one wild population and two hatchery groups to evaluate the potential impacts of hatchery release on the genetic structure of the wild population. Two different models indicated strong evidence of recent bottlenecks in all groups. The hatchery groups were lower in the mean number of alleles per locus, allelic richness, and allelic diversity compared with the wild population. The 80% membership coefficient indicated that 14% of the wild fish could be assigned as hybrids of wild and hatchery fish. Our results suggested that hatchery release will further reduce the natural genetic diversity in the wild population, change the genetic structure of the rock carp population, and may not benefit restoration of this vulnerable fish species.     

Genetic evaluation of interindividual relatedness for broodstock management of the rare species barfin flounder Verasper moseri using microsatellite DNA markers

ABSTRACT:   The barfin flounder has been recently declared to be rare, as the number of wild individuals in Japan has greatly reduced since 1975. The Japanese National Center for Stock Enhancement started the stocking program for this species on 1987. In the present study, microsatellite DNA loci were applied in order to determine the pedigree of the hatchery-produced juveniles for stock enhancement. Additionally, using six polymorphic microsatellite markers, the accuracy to assign dyads to categories of kinship was estimated using the relatedness estimator ( R _XY), similarity indexes (individual genetic identity [GI] and proportion of shared alleles [ P _S]), identification of kin groups using unweighted pair group method with arithmetic mean (UPGMA) clustering based on interindividual genetic distances (1- R _XY, 1- P _S, and 1-GI), a likelihood ratio approach, and partition methods to separate individuals into cohorts. The results were compared with the pedigree relationship previously obtained from parentage analysis. Estimation of the kinship relationship between individuals could be better inferred by means of UPGMA dendrogram clustering based on 1-GI or 1- P _S as interindividual genetic distances, or using computer software to perform partitioning of set of individuals.     

Genetic diversity of red sea bream Pagrus major in western Japan in relation to stock enhancement

SUMMARY: Stock enhancement is used in Japan as a tool to help the replenishment of wild populations of red sea bream Pagrus major . In this study, we analyzed the genetic diversity and composition of wild red sea bream at seven locations around Shikoku Island, South-west Japan, using three microsatellite loci. This analysis was done to test the hypothesis that: (i) red sea bream comprises a single Mendelian population along Japan; and (ii) stock enhancement programs around Shikoku Island are causing genetic differentiation among wild stocks. The results indicated that some locations from the Shikoku area were not significantly different from the rest of Japan, supporting the hypothesis of a single Mendelian population. Significant departures from Hardy–Weinberg equilibrium and significant pairwise F_ST among locations indicated genetic instability within this region. We suggest that the stock enhancement programs made in the region are the possible cause of this genetic instability. A management scheme for the hatcheries involved in the stock enhancement of red sea bream is presented.     

Impacts of in-stream acclimatization in post-stocking behaviour of European grayling in a Swedish stream

Abstract The post-stocking behaviour of hatchery-reared juvenile European grayling, Thymallus thymallus (L.), with in-stream acclimatization in a pool for 4, 8, and 12 days before release was compared with fish released directly into a 370-m long seminatural stream during 1995 and 1996. The behaviour of these groups was compared with fish acclimatized for 16 h in a holding pen (0.7 m³) in a more fast-flowing part of the stream. All fish were released in the experimental stream at the same time. Initially groups of 30 individuals were compared, but this was reduced because of the mortality during acclimatization. Temporal and spatial distribution, feed intake in relation to sex and size of fish were followed for 7 days after release. Between 33 and 59% of all groups of fish released into the pool left the experimental area and were recaptured in a trap at the downstream end of the experimental stream. The acclimatization period had no significant influence on the percentage of fish moving downstream. Significantly more grayling left the stream from the holding pen (76.7%) than fish released in the pool (47.2%). Of the recaptured fish, 77% had been feeding. Fish recaptured within a week had lost more than 10% of their initial weight measured at the hatchery. Fish recaptured more than 2 weeks after release had lost a significantly smaller percentage of their initial weight compared with fish recaptured in the first 2 weeks. No differences were found in spatial distribution or feeding activity between sexes.     

Genetic structure and genetic relatedness of a hatchery stock of Senegal sole (Solea senegalensis) inferred by microsatellites

Solea senegalensis is a fish species in an early stage of domestication. A commercial hatchery in southern Spain experienced a dramatic failure in spawning and embryo survival, after incorporation into the broodstock of fish reared in their hatchery. To assess the impact of management on the stock quality, a population genetics study was made of adult broodstock present at this hatchery in 2000. The broodstock was composed of a group of fish of wild origin and two more groups from F₁ progeny. A set of eight microsatellite loci was used to compare genetic variability among the three groups and to establish the relationships between pairs of individuals within each group.Individuals from F₁ origin showed a substantial reduction in genetic variability when compared to those from the wild. There was a greater than 50% decrease in the number of alleles per locus corrected by sample size and 16% and 26% reductions in H_e values in the two F₁ groups, respectively. The degree of genetic relationship between individuals, evaluated by the relatedness estimator, rxy, and by graphical approaches, revealed a high proportion, almost 75%, of full-sibs and half-sibs in both F₁ groups, which might account for the reduction in overall variability. The high proportion of siblings also suggests a reproductive pattern in this stock in which few individuals breed successfully. This mating behaviour, along with factors inherent to selection and management systems, might be responsible for the negative effects on the reproductive capacity of the stock.     

The enhancement of abalone stocks: lessons from Japanese case studies

The dramatic declines in abalone Haliotis spp. fishery production have been documented all over the world. Release of hatchery‐reared juveniles into natural habitats has been considered as one measure to sustain and/or augment the current fishery production of abalone, as well as to restore collapsed abalone stocks. However, attempts at abalone release programmes have only been undertaken at experimental scales, except for Japan, where large‐scale stock enhancement programmes for abalone have been undertaken since late 1960s. To evaluate the potential of stock enhancement for abalone, we analysed the release surveys of 13 case studies in Japan in terms of the overall recapture rate (number of recaptures through a lifetime/number of juveniles released), yield per release (YPR, yields from released individuals), the economic efficiency of releases (ratio of income from recaptured abalone to release cost) for each release year, and the contribution of hatchery releases to total catches for each fishing year. The average estimates for overall recapture rates (0.014–0.238) and YPR (3.1–60.3 g/individual) varied between locations and release years. The economic efficiency was estimated at 0.4–6.2. The released abalone contributed 6.9–83.5% to total catches. Hatchery releases could augment total production at some locations, but the success of release programmes would be limited by the carrying capacity at release areas, because density‐dependent mortality occurred following releases in some cases. Throughout Japan, the annual catch of abalone has continuously declined from ～6500 t in 1970 to ～2000 t in the mid‐1990s, despite the increase in the number of hatchery releases. Based on the estimates for YPR, the magnitude of the abalone releases on a national scale has not been sufficiently large to sustain the total production of Japanese abalone, which has primarily fluctuated according to the abundances of wild populations. Our results suggest that releases should be targeted at local populations in regions where stock enhancement is predicted to have the greatest chance of success, and the magnitude of releases should be considered carefully and determined for each region by taking the local carrying capacity into account. We also address the future prospects of abalone stock enhancement.