Differential Reproductive Success of Sympatric, Naturally Spawning Hatchery and Wild Steelhead, Oncorhynchus mykiss

Hatchery propagation of salmonids has been practiced in western North America for over a century. However, recent declines in wild salmon abundance and efforts to mitigate these declines through hatcheries have greatly increased the relative abundance of fish produced in hatcheries. The over-harvest of wild salmon by fishing mixed hatchery and wild stocks has been of concern for many years but genetic interactions between populations, such as hybridization, introgression and outbreeding depression, may also compromise the sustainability of wild populations. Our goal was to examine whether a newly established hatchery population of steelhead trout successfully reproduced in the wild and to compare their rate of reproductive success to that of sympatrically spawning native steelhead. We used eight microsatellite loci to create allele frequency profiles for baseline hatchery and wild populations and assigned the smolt (age 2) offspring of this parental generation to a population of origin. Adults originating from a generalized hatchery stock artificially selected for early return and spawning date were successful at reproducing in Forks Creek, Washington. Although hatchery females (N = 90 and 73 in the two consecutive years of the study) produced offspring that survived to emigrate as smolts, they produced only 4.4–7.0% the number produced per wild female (N = 11 and 10). This deficit in reproductive success implies that the proportion of hatchery genes in the mixed population may diminish since deliberate releases into the river have ceased. This hypothesis is being tested in a long-term study at Forks Creek.     

A Comparison of Genetic Variation Between an Anadromous Steelhead, Oncorhynchus mykiss, Population and Seven Derived Populations Sequestered in Freshwater for 70 Years

In 1926 cannery workers from the Wakefield Fisheries Plant at Little Port Walter in Southeast Alaska captured small trout, Oncorhynchus mykiss, from a portion of Sashin Creek populated with a wild steelhead (anadromous O. mykiss) run. They planted them into Sashin Lake which had been fishless to that time and separated from the lower stream by two large waterfalls that prevented upstream migration of any fish. In 1996 we sampled adult steelhead from the lower creek and juvenile O. mykiss from an intermediate portion of the creek, Sashin Lake, and five lakes that had been stocked with fish from Sashin Lake in 1938. Tissue samples from these eight populations were compared for variation in: microsatellite DNA at 10 loci; D-loop sequences in mitochondrial DNA; and allozymes at 73 loci known to be variable in steelhead. Genetic variability was consistently less in the Sashin Lake population and all derived populations than in the source anadromous population. The cause of this reduction is unknown but it is likely that very few fish survived to reproduce from the initial transplant in 1926. Stockings of 50–85 fish into five other fishless lakes in 1938 from Sashin Lake did not result in a similar dramatic reduction in variability. We discuss potential explanations for the observed patterns of genetic diversity in relation to the maintenance of endangered anadromous O. mykiss populations in freshwater refugia.     

Effects of Hatchery Rearing on Brain Structures of Rainbow Trout, Oncorhynchus mykiss

In this study, we contrast brain morphology from hatchery and wild reared stocks to examine the hypothesis that in salmonid fishes, captive rearing produces changes in brain development. Using rainbow trout, Oncorhynchus mykiss, as a model, we measured eight regions of the salmonid brain to examine differences between wild and hatchery reared fish. We find using multiple analysis of covariance (MANCOVA), analysis of covariance (ANCOVA) and discriminant function analysis (DFA) that the brains of hatchery reared fish are relatively smaller in several critical measures than their wild counterparts. Our work may suggest a mechanistic basis for the observed vulnerability of hatchery fish to predation and their general low survival upon release into the wild. Our results are the first to highlight the effects of hatchery rearing on changes in brain development inbreak fishes.     

虹鳟6个养殖群体遗传多样性的微卫星分析

利用14对微卫星分子标记对虹鳟的6个养殖群体进行遗传多样性分析。结果表明：6个群体的平均等位基因数A为 2.89～4.22,平均有效等位基因数Ne 为2.15～2.78,平均观察杂合度Ho 为0.4801～0.6786,平均期望杂合度He 为0.5052～0.6211,平均多态信息含量PIC 为0.4298～0.5762;其中渤海站群体的等位基因数最多、多态信息含量最高,有效种群最大,通过基因型的P值发现6个群体只在位点AF375034符合Hardy-Weinberg平衡,在其他位点都不同程度的偏离平衡,同时对6个群体的遗传距离进行了估算,聚类分析发现挪威群体与其他群体遗传距离最远。     

The Mating System of Steelhead, Oncorhynchus mykiss, Inferred by Molecular Analysis of Parents and Progeny

The development of molecular markers has allowed behavioral ecologists to link parents to specific offspring, providing insights into breeding systems that were not apparent from direct observations of the social system. Studies of this type in fishes have focused on species with male parental care such as centrarchids, and on salmonids, a family with little parental care. In order to gain further insight into the mating system of steelhead trout, Oncorhynchus mykiss, a winter-spawning species whose reproductive system is poorly known, adults returning to spawn were captured in four consecutive years in a small, unfished, wild population. Juvenile offspring were sampled by electrofishing and parentage was determined by exclusion based on a 12 locus microsatellite genotype. Both males and females mated with multiple individuals, though single pair matings were also inferred. Females and males tended to have the same number of mates (median = 1), but males were more likely to have no apparent partner (43% vs. 23% for females) and the maximum number of mates were obtained by males (range 0–10 vs. 0–5 for females). There was no difference in median arrival date by sex, but 80% of the females mated with males that had already arrived rather than males arriving with or after the females (median = 7.5, range = 1–63 days difference). Contrary to expectations, there was no evidence of size-assortative mating; larger males and larger females did not tend to mate with each other more often than would have occurred by chance. Of the juveniles with only one identified parent, most had a known mother and an unknown father rather than the reverse (88% vs. 11%). We interpret this as indirect evidence that non-anadromous males achieved a significant number of fertilizations. Thus the steelhead mating system was complex, being more strongly structured by arrival date than fish size, and including a significant genetic contribution by mature male parr.     

Temporal Genetic Variance and Propagule-Driven Genetic Structure Characterize Naturalized Rainbow Trout (Oncorhynchus mykiss) from a Patagonian Lake Impacted by Trout Farming

Knowledge about the genetic underpinnings of invasions—a theme addressed by invasion genetics as a discipline—is still scarce amid well documented ecological impacts of non-native species on ecosystems of Patagonia in South America. One of the most invasive species in Patagonia’s freshwater systems and elsewhere is rainbow trout (Oncorhynchus mykiss). This species was introduced to Chile during the early twentieth century for stocking and promoting recreational fishing; during the late twentieth century was reintroduced for farming purposes and is now naturalized. We used population- and individual-based inference from single nucleotide polymorphisms (SNPs) to illuminate three objectives related to the establishment and naturalization of Rainbow Trout in Lake Llanquihue. This lake has been intensively used for trout farming during the last three decades. Our results emanate from samples collected from five inlet streams over two seasons, winter and spring. First, we found that significant intra- population (temporal) genetic variance was greater than inter-population (spatial) genetic variance, downplaying the importance of spatial divergence during the process of naturalization. Allele frequency differences between cohorts, consistent with variation in fish length between spring and winter collections, might explain temporal genetic differences. Second, individual-based Bayesian clustering suggested that genetic structure within Lake Llanquihue was largely driven by putative farm propagules found at one single stream during spring, but not in winter. This suggests that farm broodstock might migrate upstream to breed during spring at that particular stream. It is unclear whether interbreeding has occurred between “pure” naturalized and farm trout in this and other streams. Third, estimates of the annual number of breeders (N_b) were below 73 in half of the collections, suggestive of genetically small and recently founded populations that might experience substantial genetic drift. Our results reinforce the notion that naturalized trout originated recently from a small yet genetically diverse source and that farm propagules might have played a significant role in the invasion of Rainbow Trout within a single lake with intensive trout farming. Our results also argue for proficient mitigation measures that include management of escapes and strategies to minimize unintentional releases from farm facilities.     

Hatchery performance in a major rainbow trout (Oncorhynchus mykiss,Walbaum, 1792) seed production area of Turkey

This study aims to determine the production characteristics and performance of rainbow trout hatcheries in Seydikemer, Mu?la, Turkey where over half of the total production of eyed eggs and juveniles take place. For this purpose six hatcheries with different production scales were selected and coded A, B, C, D, E and F (with capacities of 60, 15, 5.6, 4, 0.95 and 0.6 million juveniles per year, respectively). From each hatchery, 20 females were selected and their egg qualities (egg number, diameter and weight, fertilization, eyed egg, hatching, swim‐up fry and survival rates at 120 days after hatching) and juvenile growth rates were monitored until the 120^th day after hatching under each farms own conditions. The sperm characteristics of 10 broodstock males from each hatchery were also examined. Although fertilization and eyed egg rates were similar among the farms, hatching, swim‐up fry, and survival rates at the 120^th day after hatching differed significantly. The average eyed egg, hatching and survival rates (calculated from selected 20 females) were 72, 55, and 32%, respectively. While sperm characteristics except duration of motility (s), were significantly different among the farms, average sperm concentration (per ml), motility (%) and duration of motility (s) in the region were 11.8 × 10⁹, 55.3, and 56.4, respectively. The best hatchery performance was observed in Farm A because of better records and management applications than in the other farms.     

Effects of temperature on sexual development in steelhead,Oncorhynchus mykiss

We evaluated how genotype (stock source, parental line, chromosomal sex) and temperature interact to influence gonad development and phenotypic sex of Oncorhynchus mykiss. Embryos were generated from single pair matings of natural-origin and first-generation hatchery-origin O. mykiss held at ambient or elevated (5 °C above ambient) water temperature. Exposure to elevated temperature lasted from fertilization until the onset of exogenous feeding, after which all experimental groups were maintained at ambient temperature. Sexual phenotype was determined by histology and sexual genotype by the OmyY1 marker. There was 99% concordance among fish sexed both phenotypically and genotypically, demonstrating that experimental factors, including elevated rearing temperatures, did not result in sex change. Approximately 10% of the histologically examined fish showed no evidence of gonad development beyond the indifferent stage. Most of these fish were from hatchery family lines.

     

Molecular Markers for Variation in Spawning Date in a Hatchery Population of Rainbow Trout (Oncorhynchus mykiss)

We examined the distribution of alleles at 63 microsatellite loci distributed across 29 linkage groups in broodstock females from a commercial population of rainbow trout spawning on different dates throughout the season (August to January). A total of 368 females, 184 and 117 females from each of the tail-ends of the spawning distribution and a subsample of 67 females spawning in the middle, were used to detect marker–trait associations. Twenty-one loci in a subset of genomic regions (RT-5, 7, 8, 10, 12, 14, 15, 22, 23, 24, 25, 29, 30, and 31) were significantly associated with variation in spawning date. Many of these markers localize to regions with known spawning date quantitative trait loci based on previous studies. An individual assignment analysis was used to test how well the molecular data could be used to assign individuals to their correct spawning group, and markers were given a ranking reflecting their contribution to the accuracy of assignment. The top 15 ranked markers were successful at assigning the majority of females to the correct spawning group based on genotype with an average accuracy of 76 %. The most likely genes that could contribute to these differences in spawning date are discussed. Together, these data indicate that the loci could be incorporated into a selection index with phenotype data to increase the accuracy of selection for spawning date.     

新疆桑属植物栽培居群的遗传多样性研究

应用RAPD分子标记对新疆不同地区栽培的桑属植物2种3个分类群共11个居群进行了遗传多样性研究。结果表明,新疆桑属栽培植物中虽然存在较为丰富的遗传多样性,多态位点比率(PPB)为87．39％,Shannon多样性指数为0．3997,但在栽培居群内的遗传变异水平相对较低;在不同居群间遗传变异水平仔住很人差异,各居群的多态位点比率(PPB)为4．5％至45．95％,Shannon多样性指数为0．0312至0．2339;白桑(Morus alba L．)及其变种鞑靼桑(Morus alba L．var．tatarica)居群内的遗传变异水平远高于黑桑种(Morus nigra)。新疆桑属植物栽培居群内较低的遗传变异水平与其采用扦插等无性繁殖方式有关。分析全部的遗传变异显示,11个栽培居群之间的基因分化系数(Gst)为0．3541,其中桑及其变种9个居群间的基因分化系数为0．4597,黑桑种2个居群问的基因分化系数为0．4728。AMOVA分析表明,在全部遗传变异中,黑桑种和白桑种2个物种之间的遗传变异占59．16％,居群间遗传变异为17．46％。遗传距离和聚类分析也表明,黑桑种和白桑种及其变种鞑靼桑之间存在很大的遗传分化。     

Genetic Diversity of Orobanche cumana Populations in Serbia

In this study, we report genetic characterization of Orobanche cumana, the causal agent of sunflower wilting in Serbia. The genetic diversity of this parasitic plant in Serbia was not studied before. Random amplified polymorphic DNA (RAPD) markers and partial rbcL gene sequences analysis were used to characterize the O. cumana populations at the molecular level. While phylogenetic analyses of RAPD-PCR amplicons were performed using unweighted pair-group Method analyses, rbcL gene sequences were analyzed using neigbor joining method and minimum spanning tree. Molecular analyses of RAPD-PCR analysis revealed high genetic diversity of O. cumana populations which indicated high adaptive potential of this parasitic weed in Serbia. Further analyses of rbcL gene using minimum spanning tree revealed clear differences among diverse sections of Orobanche genus. Although this molecular marker lacked the resolution to display intrapopulation diversity it could be a useful tool for understanding the evolution of this parasitic plant. Our results suggested that O. cumana has great genetic potential which can lead to differentiation of more virulent races which is important for determining crop breeding strategies for their control.     

The Spring-Run Ecotype of the Steelhead Trout Parasalmo (Oncorhynchus) mykiss (Walbaum, 1792) (Salmonidae,Salmoniformes) in the Kamchatka Peninsula

Russian Journal of Marine Biology - A seasonal spring-run (vernal) ecotype of the Steelhead Trout Parasalmo (Oncorhynchus) mykiss (Walbaum, 1792) has been found in western Kamchatka for the first...     

Temporal and spatial variability in nearshore bacterioplankton communities of Lake Michigan

The spatial and temporal variability of bacterial communities were determined for the nearshore waters of Lake Michigan, an oligotrophic freshwater inland sea. A freshwater estuary and nearshore sites were compared six times during 2006 using denaturing gradient gel electrophoresis (DGGE). Bacterial composition clustered by individual site and date rather than by depth. Seven 16S rRNA gene clone libraries were constructed, yielding 2717 bacterial sequences. Spatial variability was detected among the DGGE banding patterns and supported by clone library composition. The clone libraries from deep waters and the estuary environment revealed highest overall bacterial diversity. Betaproteobacteria sequence types were the most dominant taxa, comprising 40.2–67.7% of the clone libraries. BAL 47 was the most abundant freshwater cluster of Betaproteobacteria , indicating widespread distribution of this cluster in the nearshore waters of Lake Michigan. Incertae sedis 5 and Oxalobacteraceae sequence types were prevalent in each clone library, displaying more diversity than previously described in other freshwater environments. Among the Oxalobacteraceae sequences, a globally distributed freshwater cluster was determined. The nearshore waters of Lake Michigan are a dynamic environment that experience forces similar to the coastal ocean environment and share common bacterial diversity with other freshwater habitats.     

Effects of Desacetyl-alpha-MSH on Lipid Mobilization in the Rainbow Trout, Oncorhynchus mykiss

Effects of melanocyte-stimulating hormone (MSH) and beta-endorphin on lipid mobilization were examined in the rainbow trout (Oncorhynchus mykiss). Plasma levels of fatty acid (FA) were measured after intra-arterial administration of alpha-MSH, desacetyl-alpha-MSH, beta-MSH, or beta-endorphin through a cannula in the dorsal aorta. Desacetyl-alpha-MSH at 1 ng/g body weight resulted in an increase in plasma FA levels 1-3 hr after the injection, whereas the other three peptides showed no significant effect at the same dose. There was no significant change in plasma levels of cortisol after administration of any of the peptides. Lipolytic enzyme activity in the liver was significantly increased in a dose-related manner 1 hr after single intra-peritoneal injection of desacetyl-alpha-MSH. The direct effect of desacetyl-alpha-MSH on lipolysis was examined in liver slices incubated in vitro. Lipase activity in the liver slice was stimulated in the medium containing desacetyl-alpha-MSH in a dose-related manner. The results indicate that desacetyl-alpha-MSH is a potent stimulator of lipid mobilization in the rainbow trout.     

缢蛏种群遗传多样性的周年变异分析

利用线粒体COI标记分析了福建省宁德市漳湾镇横屿村滩涂5个月份缢蛏群体(S3,S5,S7,S9,S11)的遗传结构,以期评估不同月份时期种群的遗传多样性差异。基于线粒体COI基因的结果表明,5个缢蛏群体的平均核苷酸差异数位于2.7836-3.6894之间,核苷酸多样性指数位于0.0050-0.0066之间,遗传多样性水平大致表现为S3和S5群体较高于S7和S9群体,明显高于S11群体。AMOVA分析结果显示,群体间遗传变异量占总变异的7.18%,而群体内变异达到了92.82%,说明遗传差异主要来自于群体内部。由此可见,从3月份到11月份,缢蛏群体的遗传多样性水平呈现出下降趋势,尤其是在11月份,差异最为明显。     

西南地区硬枝野荞麦(Fagopyrum urophyllum)居群的遗传多样性研究

采用等位酶电泳技术了云南省中北部昆明、富民、宾川3县（市）及四川省西南布拖县的硬枝野荞麦（Fagopyrum urophyllum)6个天然居群的遗传多样性和分化。硬枝野荞麦居群内维持有较高的遗传多样性,多态位点比率为50．0％,预期杂合度和观察杂合度分别为0．251和0．471。并对硬枝野荞麦（F.urophyllum)与栽培荞麦之间遗传变异作了比较。     

浙江省境内乌药种群遗传多样性研究

利用ISSR分子标记方法对浙江省境内的8个乌药自然种群的遗传多样性和遗传分化进行了研究。从100个引物中筛选出12个用于正式扩增的ISSR引物,在8个种群160个个体中共检测到125个位点,其中多态位点100个,多态位点百分率(P)为80%,各种群P平均为60.5%。乌药总的Shannon信息指数(I)为0.3762,Nei指数(h)为0.246 4,各种群I平均为0.317 3,h平均为0.213 1。AMOVA分析表明在总的遗传变异中,11.44%的变异发生在种群间,88.56%的变异发生于种群内,种群间的遗传分化系数(Gst)为0.134 9,乌药种群间的基因流(Nm)为3.205 2。8个种群间的平均遗传距离为0.049 4,利用算术加权平均数法(UPGMA)对乌药8个种群进行聚类,结果可分为2大类群,台州市的水磨坑、上泮、苍山等3个种群聚为一类,其余5个种群聚为另一类。     

滇牡丹天然居群的遗传多样性分析

以云南中部及西北部的6个滇牡丹(Paeonia delavayi)天然居群为研究对象,进行株高、新枝长等9个表型性状的表型多样性分析和ISSR分析。结果表明:9个表型性状变异幅度为0.9%~39.8%,平均值达到了18.9%;居群间生殖器官的变异较大,居群内营养器官更容易产生变异。利用居群间欧式距离进行聚类分析,6个居群聚为4个类群,没有与实际地理位置相吻合,说明表型特征的性状与地理距离的相关性不大。遗传多样性分析结果表明:利用筛选得到的10条引物,在取自6个自然居群、180个个体中,检测到56个多态位点。在居群水平上,多态位点百分率(PPB)为60.2%,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.281和0.414。在物种水平上,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.409和0.596。居群间的遗传分化系数(Gst)达0.319。结果显示,表型性状在居群间和居群内均存在广泛变异。滇牡丹遗传多样性水平较高,居群间遗传分化较大,滇牡丹并不濒危。     

内蒙古布氏田鼠地理种群遗传多样性

以MHCⅡ类基因第二外显子为分子标记,利用限制性内切酶分析法和序列分析法对8个布氏田鼠Lasiopodomys brandtii地理种群进行遗传多样性分析。结果显示,8个布氏田鼠地理种群的MHCⅡ类基因第二外显子酶切共检测到6个等位基因,定义21种单倍型,其中有3个单倍型为不同区域种群共享,经卡方检验,6个酶切多态性位点上基因型频率不符合Hardy-Weinberg平衡;序列分析显示,在261 bp的核苷酸序列中,有57个变异位点,单倍型多样性为0.746 5~0.873 3、核苷酸多样性为0.006 06~0.016 55。谱系分析得到3个稳定的分支:锡林浩特、二连浩特、东乌珠穆沁旗和西乌珠穆沁旗4个种群形成一个单元分歧的进化分支(A区域),新巴尔虎左旗、陈巴尔虎旗和新巴尔虎右旗3个种群成一个单元分歧的进化分支(B区域),而位于浑善达克沙漠南部的正镶白旗种群形成一个独立单元分歧的进化分支(C区域),分别与采集的地理种群相吻合。AMOVA分析结果表明,区域类群间的遗传变异占总变异比率的64.08%,区域内种群间占7.78%,种群内占28.14%。除正镶白旗种群外,布氏田鼠种群具有较丰富的遗传多样性,遗传变异主要发生在区域类群之间和种群内。     

Effects of experimental anaemia on intra-erythrocytic phosphate levels in rainbow trout, Oncorhynchus mykiss

The concentrations of intra-erythrocytic adenylates (ATP, ADP and AMP) and guanylates (GTP, GDP and GMP) were determined in rainbow trout subjected to 10% blood removal every 12 h for 96 h. Haemoglobin concentration, [Hb], decreased from 6·043±0·617 to 0·957 ± 0·195 g dl⁻¹. This decrease in [Hb] was followed by a continuous increase in total organic phosphates, e.g. adenylates plus guanylates. Intra-erythrocytic NTP (ATP plus GTP) levels increased significantly after 48 h when haemoglobin concentration was 2·427 ± 0·256 g dl⁻¹. Although a significant increase in GDP levels in animals with [Hb] less than 1·677 ± 0·235 g dl⁻¹ was observed, the general increase in guanylate level was mainly due to the GMP which increased about 85-fold during the experimental period. It is suggested that the erythrocytes of anaemic rainbow trout have the capacity to increase NTP/Hb₄ ratios which may represent an advantage for anaemic fish.