Female Salmon Research Articles

Biological Characteristics of Fish Germ Cells and their Application to Developmental Biotechnology

We have revealed several unique characteristics of germ cell development using rainbow trout, including the fact that spermatogonia transplanted into the peritoneal cavity of newly hatched embryos migrate toward recipient gonads, that spermatogonia transplanted into female recipients start oogenesis and produce functional eggs and that diploid germ cells transplanted into triploid trout can complete gametogenesis. By combining these unique features of fish germ cells, we established allogeneic and xenogeneic transplantation systems for spermatogonia in several fish species. Spermatogonia isolated from the mature testes of vasa-green fluorescent protein (Gfp) transgenic rainbow trout were transplanted into the peritoneal cavity of triploid masu salmon newly hatched embryos. These spermatogonia migrated toward recipient salmon genital ridges with extending pseudopodia and were subsequently incorporated into them. We further confirmed that the donor-derived spermatogonia resumed gametogenesis and produced sperm and eggs in male and female salmon recipients, respectively. By inseminating the resulting eggs and sperm, we obtained only rainbow trout offspring in the F1 generation, suggesting that the triploid salmon recipients produced functional gametes derived only from donor trout. We further confirmed that this intra-peritoneal transplantation of germ cells is applicable to several marine fishes, which could be of benefit in the production of bluefin tuna that has a large broodstock (>100 kg) and is difficult to maintain in captivity. Gamete production of bluefin tuna could be more easily achieved by generating a surrogate species, such as mackerel, that can produce tuna gametes.

Consequences of acute stress and cortisol manipulation on the physiology, behavior, and reproductive outcome of female Pacific salmon on spawning grounds

Life-history theory predicts that stress responses should be muted to maximize reproductive fitness. Yet, the relationship between stress and reproduction for semelparous salmon is unusual because successfully spawning individuals have elevated plasma cortisol levels. To tease apart the effects of high baseline cortisol levels and stress-induced elevation of cortisol titers, we determined how varying degrees of cortisol elevation (i.e., acute and chronic) affected behavior, reproductive physiology, and reproductive success of adult female pink salmon (Oncorhynchus gorbuscha) relative to different states of ovulation (i.e., ripe and unripe). Exhaustive exercise and air exposure were applied as acute stressors to manipulate plasma cortisol in salmon either confined to a behavioral arena or free-swimming in a spawning channel. Cortisol (eliciting a cortisol elevation to levels similar to those in post-spawn female salmon) and metyrapone (a corticosteroid synthesis inhibitor) implants were also used to chemically manipulate plasma cortisol. Cortisol implants elevated plasma cortisol, and impaired reproductive success; cortisol-treated fish released fewer eggs and died sooner than fish in other treatment groups. In contrast, acute stressors elevated plasma cortisol and the metyrapone implant suppressed plasma cortisol, but neither treatment significantly altered reproductive success, behavior, or physiology. Our results suggest that acute stressors do not influence behavior or reproductive outcome when experienced upon arrival at spawning grounds. Thus, certain critical aspects of salmonid reproduction can become refractory to various stressful conditions on spawning grounds. However, there is a limit to the ability of these fish to tolerate elevated cortisol levels as revealed by experimental elevation of cortisol.

Tradeoffs between homing and habitat quality for spawning site selection by hatchery-origin Chinook salmon

Spawning site selection by female salmon is based on complex and poorly understood tradeoffs between the homing instinct and the availability of appropriate habitat for successful reproduction. Previous studies have shown that hatchery-origin Chinook salmon (Oncorhynchus tshawytscha) released from different acclimation sites return with varying degrees of fidelity to these areas. To investigate the possibility that homing fidelity is associated with aquatic habitat conditions, we quantified physical habitat throughout 165 km in the upper Yakima River basin (Washington, USA) and mapped redd and carcass locations from 2004 to 2008. Principal components analysis identified differences in substrate, cover, stream width, and gradient among reaches surrounding acclimation sites, and canonical correspondence analysis revealed that these differences in habitat characteristics were associated with spatial patterns of spawning (p < 0.01). These analyses indicated that female salmon may forego spawning near their acclimation area if the surrounding habitat is unsuitable. Evaluating the spatial context of acclimation areas in relation to surrounding habitat may provide essential information for effectively managing supplementation programs and prioritizing restoration actions.

Seasonal changes in depth distribution of salmon sharks (Lamna ditropis) in Alaskan waters: implications for foraging ecology

The salmon shark, Lamna ditropis, is an endothermic, apex predator in the productive waters of the Gulf of Alaska (GOA). To understand their role in these productive habitats and to identify and better understand their interactions with commercially important species and fisheries, detailed information regarding the distribution of the salmon shark is required. Archival data records from eight recovered pop-up archival transmitting (PAT) tags were analyzed to examine the geographic and vertical distribution of female salmon sharks and elucidate how their distribution changes in relation to the biotic and abiotic environment. Tagged sharks used the neritic habitats of the GOA for the duration of the PAT deployments. Sharks exhibited a diel periodicity in their dive behavior. Salmon sharks exhibited three different patterns of depth distribution (behavioral modes) that occurred during distinct oceanographic seasons, likely reflecting changes in their foraging ecology in response to seasonal changes in the distribution and availability of important prey. The distribution of salmon sharks in the GOA appears to follow consistent patterns and has a high degree of geographical and vertical overlap with commercially important prey species. This information increases our understanding of interactions between salmon sharks and commercial fisheries and may assist in fostering responsible management for this species.

Reproductive timing phenotypes in female salmon: true alternatives or extreme variants?

Kokanee salmon, Oncorhynchus nerka, can vary in their maturation status at arrival to spawning areas and in their subsequent behaviour. In Meadow Creek, British Columbia, typical ‘red-arriving’ females arrive with red nuptial coloration, spawn within a few days, and then defend their nests against digup by conspecifics. In contrast, atypical ‘silver-arriving’ females arrive with scales still present and delay maturation and spawning by 3–4 weeks. Our objective was to further describe the silver- and red-arriving phenotypes and to test the hypothesis that they are proxies for a dimorphism of life cycle variation. Colour was a good indicator of a female’s stage of maturity and senescence because silver-arriving females had larger guts, larger and fattier livers, and smaller eggs containing less lean mass than red-arriving females. Colour, and therefore maturity status, covaried with life history traits: silver-arriving females were younger (in 2008 and 2009) and smaller (in 2008) than red-arriving females. The percentage of silver-arriving females was higher in 2009 (11.4%) than in 2008 (2.2%), possibly because of poor conditions for growth in 2009. We also observed some silver-arriving males, but they were much rarer (0.4% in 2008 and 1.5% in 2009). Scores on the first discriminant function based on colour information from digital photographs distinguished silver-arriving and red-arriving females. At the population level, these scores were bimodally distributed, but the dip between modes did not clearly separate silver- and red-arriving females. Although silver- and red-arriving phenotypes represent female reproductive tactics for delivering parental care, they are not distinct alternatives and instead represent the extremes of life cycle variation in this population.

Maintenance of asymmetric hybridization between Atlantic salmon (Salmo salar) and brown trout (Salmo trutta) via postzygotic barriers and paternal effects

At the southern European edge of Atlantic salmon ( Salmo salar ) distribution, all the hybrids found in nature are the product of crosses between female salmon and male brown trout ( Salmo trutta ). By artificially producing reciprocal crosses between salmon and trout, we demonstrate that unidirectional hybridization observed in nature is the result of postzygotic barriers that produce very high mortality rates (95%) in female trout × male salmon hybrids and not the consequence of prezygotic isolation or behavioural differences between the two species. Mortality of female trout × male salmon hybrids mainly occurs during the last phases of development, and a high percentage of these surviving hybrids showed external deformities that could compromise survival in the wild. Another important finding is the existence of paternal factor in embryo development. Using time to midhatch as an indicator of developmental rate, female salmon × male trout hybrids hatched faster than female trout × male salmon hybrids, with both developing at a rate intermediate to the pure crosses. The early emergence of female salmon × male trout hybrids, which have similar survival to pure salmons, could have fitness repercussions, since early emerging fry have a competitive advantage over later emerging fry.

Tissue residue concentrations of organohalogens and trace elements in adult Pacific salmon returning to the Fraser River, British Columbia, Canada

We report measured concentrations of organohalogens and trace elements in muscle and eggs of returning wild Pacific sockeye and chinook salmon during their 2007 migration through the Fraser River watershed in Canada. Chemical analyses revealed the presence of ppb to ppm levels of a wide variety of contaminants in these fish, including polychlorinated biphenyls (PCBs); polychlorinated dibenzo-p-dioxins (PCDDs); polychlorinated dibenzofurans (PCDFs); polybrominated diphenyl ethers (PBDEs); organochlorine pesticides (OCPs) such as DDTs, hexachlorocyclohexanes (HCHs), octachlorostyrene, and cyclodienes; and Hg, As, Cd, Pb, and several other trace elements. Body weights and flesh lipid contents declined during upstream migration, resulting in significantly higher (p < 0.05) lipid-normalized concentrations of lipophilic organohalogens (PCBs, PCDD/Fs, pesticides) in those spawning salmon. Postmigration magnification factors (MFs) of organohalogens (0.1-10) were comparable to previous observations and model predictions. MFs generally increased with increasing hydrophobicity (K(OW)). For example, MFs of tetra- and pentachlorobenzenes and HCH isomers (log K(OW) range: 3.8-5) were relatively low (between 0.1 and 1.7) compared with those of more lipophilic compounds (log K(OW) > 6) such as PCBs, DDTs, and mirex (MFs between 5 and 10). Lipid-normalized muscle:egg ratios in female salmon, which varied between 0.1 and 8, also exhibited a positive relationship with chemical K(OW). The results indicate that lipophilic compounds (K(OW) > 10(6)) can be magnified in flesh lipids of Pacific salmon during spawning migration, but maternal transfer kinetics (deposition to eggs) of those chemicals are relatively slow compared with less hydrophobic compounds. 2,3,7,8-TCDD toxic equivalents (ΣTEQs) in eggs of these spawning salmon, calculated using WHO toxic equivalency factors (WHO-TEFs) for fish health, in some cases exceeded the 0.3 pg·g(-1) threshold level associated with 30% salmonid egg mortality, indicating the potential for reproductive impacts in Fraser River salmon populations.

Temporally stable population-specific differences in run timing of one-sea-winter Atlantic salmon returning to a large river system.

The understanding of migration patterns can significantly contribute to conservation and management. The spawning migrations of Atlantic salmon (Salmo salar) cover thousands of kilometers from the feeding areas at sea to their natal rivers to reproduce. Migrating salmon are exposed to intensive harvest, but little is known of the population-specific differences in migration behavior. In this study, timing of return migration was investigated among one-sea-winter Atlantic salmon within a river system. By utilizing knowledge of the genetic population structure, population of origin was reliably identified for c. 1500 fish caught in mixed stock fisheries after adopting an approach to minimize the complications arising from potential nonsampled populations. Results demonstrated significant and temporally stable differences among populations as well as between sexes. Generally, female salmon from tributary populations entered fresh water first. Run timing was not however related to in-river migration distance. Rather, one-sea-winter salmon from larger populations and with a higher proportion of multi-sea-winter females arrived later in the season. These findings are a significant step toward a more thorough understanding of the salmon migration behavior and behavioral ecology, providing concrete tools for the management and conservation of the remaining indigenous Atlantic salmon stocks.

Indirect Benefits for Female Salmon from Mating with Brown Trout

By genetic analysis of 1625 samples from 10 south European rivers, we have found that Atlantic salmon Salmo salar hybridize with sympatric brown trout S. trutta in the wild and provide the female in most heterospecific crosses. Hybrids exhibit reduced fertility and could be considered a wasted reproductive effort by salmon females. In 7 experiments involving salmon females, large brown trout males, and small salmon male sneakers, reproductive success of Atlantic salmon females mating with brown trout males was not significantly different from that of 5 experiments of females mating with conspecific males because small Atlantic salmon sneakers fertilized most ova (mean 93%) in salmon x trout matings. Although egg retention tended to be higher in heterospecific than in conspecific crosses (mean 5.7% vs. 20.5% respectively), mean offspring survival was 24.4% and 30.3%, respectively (t = 1.5 x 10(-8), not significant). Brown trout males taking on a courting role may benefit late-maturing females in absence or scarcity of anadromous salmon males because they play a protective role against disturbances from other fishes (including cannibal sneakers).

Influence of European sculpin, Cottus gobio, on Atlantic salmon Salmo salar, recruitment and the effect of gravel size on egg predation – implications for spawning habitat restoration

Abstract The study examined if recruitment of juvenile Atlantic salmon, Salmo salar L., was influenced by the presence of European sculpin, Cottus gobio L., and if the spawning substrate size used by salmon influences sculpin predation on salmon eggs. A combination of information on spawning site selection by female salmon and associated densities of juvenile salmon indicated that recruitment of juvenile salmon was 10 times lower in areas where sculpin was present than areas without sculpin. Predation rate on salmon eggs was found to be dependent on substrate size in artificial redds created in stream aquaria using four different sizes of substrate (13, 23, 37 and 62 mm). Predation rate averaged 83% in the aquaria with the largest substrate size, whereas a rate of only 2–3% was observed using smaller substrate sizes. Sculpin may thus be an important factor influencing the recruitment of juvenile salmon. Selecting small enough gravel sizes during restoration of salmon spawning habitat could therefore be important to minimise egg predation.

In vitro fertilization experiments using sockeye salmon reveal that bigger eggs are more fertilizable under sperm limitation.

Although theory and widespread evidence show that the evolution of egg size is driven primarily by offspring and maternal fitness demands, an additional explanation invokes sperm limitation as a selective force that could also influence egg size optima. Levitan proposed that constraints from gamete encounter in external fertilization environments could select for enlargement of ova to increase the physical size of the fertilization target. We test this theory using in vitro fertilization experiments in an externally fertilizing fish. Sockeye salmon (Onchorhyncus nerka) females show considerable between-individual variation in ovum size, and we explored the consequences of this natural variation for the fertilization success of individual eggs under conditions of sperm limitation. By engineering consistent conditions where in vitro fertilization rate was always intermediate, we were able to compare the sizes of fertilized and unfertilized eggs across 20 fertilization replicates. After controlling for any changes in volume through incubation, results showed that successfully fertilized eggs were significantly larger than the eggs that failed to achieve fertilization. Under conditions without sperm limitation, fertility was unaffected by egg size. Our findings therefore support Levitan's theory, demonstrating empirically that some element of egg size variation could be selected by fertilization demands under sperm limitation. However, further research on sperm limitation in natural spawnings is required to assess the selective importance of these results.

Quantifying the behavioral response of spawning chum salmon to elevated discharges from Bonneville Dam, Columbia river, USA

AbstractChum salmon Oncorhynchus keta that spawn in main‐stem habitats below Bonneville Dam on the Columbia River, USA, are periodically subjected to elevated discharges that may alter spawning behaviour. We investigated behavioural responses of spawning chum salmon to increased water velocities associated with experimental increases in tailwater elevation using acoustic telemetry and a dual‐frequency identification sonar. Chum salmon primarily remained near their redds at base tailwater elevations (3.5 m above mean sea level), but displayed different movement and behavioural responses as elevations were increased to either 4.1 or 4.7 m for 8‐h periods. When velocities remained suitable (&lt;0.8 m s−1) during elevated‐tailwater tests, female chum salmon remained near their redds but exhibited reduced digging activity as water velocities increased. However, when velocities exceeded 0.8 m s−1, the females that remained on their redds exhibited increased swimming activity and digging virtually ceased. Female and male chum salmon that left their redds when velocities became unsuitable moved mean distances ranging from 32 to 58 m to occupy suitable velocities, but returned to their redds after tailwaters returned to base levels. Spawning events (i.e. egg deposition) were observed for five of nine pairs of chum salmon following tests indicating any disruptions to normal behaviour caused by elevated tailwaters were likely temporary. We believe a chum salmon's decision to either remain on, or leave, its redd during periods of unsuitably high water velocities reflects time invested in the redd and the associated energetic costs it is willing to incur. Published in 2009 by John Wiley &amp; Sons, Ltd.

Cytological and biochemical studies in back-crosses between the hybrid Atlantic salmon X sea trout and its parental species.

A male hybrid between a female salmon from Lule river in Northern Sweden and a male trout from Ireland has been back-crossed to a female salmon and a female trout both from Lule river. Eleven months after the crosses had been made no offspring was left in the combination trout × hybrid, while 4 individuals of the combination salmon × hybrid were still alive. The chromosome numbers of the fish studied are: salmon, 2n=58; trout, 2n=80; hybrid, 2n=69; back-crossed individuals 2n=60, 64 and 66. Various proteins of the different components were analysed. To our knowledge this is the first joint cytological/electrophoretic investigation of hybrids of this salmonid combination.

Thiamine Content of Eggs and Lengths of Coho Salmon (Oncorhynchus kisutch) in Relation to Abundance of Alewife (Alosa pseudoharengus) in Eastern Lake Ontario, 2003 to 2006

ABSTRACT Early mortality syndrome in fry of Great Lakes salmonines is linked to reduced levels of thiamine in eggs, which reflects maternal consumption of forage fishes such as alewife (Alosa pseudoharengus) that contain thiaminase, an enzyme that destroys thiamine. We assessed annual variations in abundance and condition of alewives and thiamine status of coho salmon (Oncorhynchus kisutch) in Lake Ontario. We analyzed total thiamine in eggs of 20 coho salmon collected annually between 2003 and 2006 at the Salmon River Hatchery on the Salmon River, New York. Alewife abundance was assessed annually in southern and eastern Lake Ontario with bottom trawls during late April and early May. Mean thlamine concentration in eggs varied annually, with those collected in 2003 (2.5 nmol/g) being significantly higher than those collected in 2004 to 2006 (1.5 to 1.7 nmol/g). Although we did not test survival of fry, if reported threshold levels of thiamine for preventing mortality of Lake Michigan coho salmon fry apply, then many or most Lake Ontario coho salmon produced fry were likely to incur thiamine-deficiency mortality, especially during years 2004 to 2006. Comparison to indices of annual abundance of alewife in Lake Ontario with thiamine concentration in coho salmon eggs failed to show any significant correlations (P > 0.05). However, total length of female spawning coho salmon was positively correlated (P < 0.05) with increasing condition and estimated energy content of adult alewives in the previous spring. These results suggest that growth of coho salmon in Lake Ontario was first limited by energy intake, whereas the amount of thiamine provided by alewives was sufficient for growth (in length) but not for producing thiamine-adequate eggs.

Migration of an upper trophic level predator, the salmon shark Lamna ditropis, between distant ecoregions

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 372:253-264 (2008) - DOI: https://doi.org/10.3354/meps07706 Migration of an upper trophic level predator, the salmon shark Lamna ditropis, between distant ecoregions Kevin C. Weng1,3,*, David G. Foley2, James E. Ganong1, Christopher Perle1, George L. Shillinger1, Barbara A. Block1 1Hopkins Marine Station, Stanford University, Pacific Grove, California 93950 USA 2Environmental Research Division, NOAA Fisheries Service, Pacific Grove, California 93950, USA 3Present address: School of Ocean and Earth Science and Technology, University of Hawaii at Manoa, Honolulu, Hawaii 96822, USA *Email: kevincmweng@gmail.com ABSTRACT: Knowledge of how animals move through heterogeneous environments is essential to understanding the ecological functions they fulfill in each habitat and their responses to environmental change. Upper trophic level organisms exert structural influences through the food web, so information on their range, migration and foraging strategy is necessary to understanding ecosystem function. Recent technological advances have enabled researchers to follow individual animals over seasonal and multi-year timescales, revealing long-distance migrations in a variety of taxa. We used satellite telemetry to monitor female salmon sharks Lamna ditropis and remote sensing to characterize their environment. Salmon sharks ranged throughout the entire eastern North Pacific Ocean during a seasonal migration cycle. During long-distance migrations, quantitative movement analyses of speed, path straightness and first passage time (FPT) revealed area-restricted search (ARS) behaviors in northern and southern regions, with transiting behaviors at mid-latitudes. Individuals migrating to a highly productive southern region displayed more ARS behaviors than those moving to a low productivity region. The combination of multi-year time-series of animal behavior with synoptic environmental data reveals factors influencing migration and indicates that different life history functions are fulfilled in each habitat. KEY WORDS: Migration · Behavior · Habitat selection · Landscape ecology · Oceanography · Foraging behavior · Reproduction · Elasmobranch · Lamna ditropis Full text in pdf format Supplementary appendix PreviousNextCite this article as: Weng KC, Foley DG, Ganong JE, Perle C, Shillinger GL, Block BA (2008) Migration of an upper trophic level predator, the salmon shark Lamna ditropis, between distant ecoregions. Mar Ecol Prog Ser 372:253-264. https://doi.org/10.3354/meps07706Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 372. Online publication date: December 09, 2008 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2008 Inter-Research.

Cloning of somatolactin alpha, beta forms and the somatolactin receptor in Atlantic salmon: Seasonal expression profile in pituitary and ovary of maturing female broodstock

BackgroundSomatolactin (Sl) is a fish specific adenohypophyseal peptide hormone related to growth hormone (Gh). Some species, including salmonids, possess two forms: Sl alpha and Sl beta. The somatolactin receptor (slr) is closely related to the growth hormone receptor (ghr). Sl has been ascribed many physiological functions, including a role in sexual maturation. In order to clarify the role of Sl in the sexual maturation of female Atlantic salmon (Salmo salar), the full length cDNAs of slr, Sl alpha and Sl beta were cloned and their expression was studied throughout a seasonal reproductive cycle using real-time quantitative PCR (RTqPCR).MethodsAtlantic salmon Sl alpha, Sl beta and slr cDNAs were cloned using a PCR approach. Gene expression of Sl alpha, SL beta and slr was studied using RTqPCR over a 17 month period encompassing pre-vitellogenesis, vitellogenesis, ovulation and post ovulation in salmon females. Histological examination of ovarian samples allowed for the classification according to the degree of follicle maturation into oil drop, primary, secondary or tertiary yolk stage.ResultsThe mature peptide sequences of Sl alpha, Sl beta and slr are highly similar to previously cloned salmonid forms and contained the typical motifs. Phylogenetic analysis of Atlantic salmon Sl alpha and Sl beta shows that these peptides group into the two Sl clades present in some fish species. The Atlantic salmon slr grouped with salmonid slr amongst so-called type I ghr. An increase in pituitary Sl alpha and Sl beta transcripts before and during spawning, with a decrease post-ovulation, and a constant expression level of ovarian slr were observed. There was also a transient increase in Sl alpha and Sl beta in May prior to transfer from seawater to fresh water and ensuing fasting.ConclusionThe up-regulation of Sl alpha and Sl beta during vitellogenesis and spawning, with a subsequent decrease post-ovulation, supports a role for Sl during gonadal growth and spawning. Sl could also be involved in calcium/phosphate mobilization associated with vitellogenesis or have a role in energy homeostasis associated with lipolysis during fasting. The up-regulation of both Sl alpha and Sl beta prior to fasting and freshwater transfer, suggests a role for Sl linked to reproduction that may be independent of the maturation induced fasting.

Combining turbine blade-strike and life cycle models to assess mitigation strategies for fish passing dams

The combined model produced a tool for evaluating effects on fish populations from passage through hydropower turbines at dams. Mean blade-strike mortality was higher for adult Atlantic salmon ( Salmo salar ) and sea-run brown trout ( Salmo trutta ) (25.2%–45.3%) than for juveniles (5.3%–9.7%). Based on life cycle modeling, salmon populations in two Swedish rivers responded differently to strategies for mitigating mortality caused by fish striking turbine blades. Although population growth rates increased up to 3% and were relatively similar for both rivers, the relative increase in the number of female salmon escaping above a dam annually after 20 years when both juveniles and adults were protected was greater in the River Piteälven (68%) than in the River Vindelälven (46%) and was approximately four times greater in the River Piteälven (38% vs. 10%) when only adults were protected. These population responses were not predicted by estimates of mortality through turbines. They showed the model could be used to evaluate strategies to conserve fish populations affected by dams located in fish migratory corridors and how postspawn adults contributed to population productivity.

Salmon louse (Lepeophtheirus salmonis) transcriptomes during post molting maturation and egg production, revealed using EST-sequencing and microarray analysis

BackgroundLepeophtheirus salmonis is an ectoparasitic copepod feeding on skin, mucus and blood from salmonid hosts. Initial analysis of EST sequences from pre adult and adult stages of L. salmonis revealed a large proportion of novel transcripts. In order to link unknown transcripts to biological functions we have combined EST sequencing and microarray analysis to characterize female salmon louse transcriptomes during post molting maturation and egg production.ResultsEST sequence analysis shows that 43% of the ESTs have no significant hits in GenBank. Sequenced ESTs assembled into 556 contigs and 1614 singletons and whenever homologous genes were identified no clear correlation with homologous genes from any specific animal group was evident. Sequence comparison of 27 L. salmonis proteins with homologous proteins in humans, zebrafish, insects and crustaceans revealed an almost identical sequence identity with all species.Microarray analysis of maturing female adult salmon lice revealed two major transcription patterns; up-regulation during the final molting followed by down regulation and female specific up regulation during post molting growth and egg production. For a third minor group of ESTs transcription decreased during molting from pre-adult II to immature adults. Genes regulated during molting typically gave hits with cuticula proteins whilst transcripts up regulated during post molting growth were female specific, including two vitellogenins.ConclusionThe copepod L.salmonis contains high a level of novel genes. Among analyzed L.salmonis proteins, sequence identities with homologous proteins in crustaceans are no higher than to homologous proteins in humans. Three distinct processes, molting, post molting growth and egg production correlate with transcriptional regulation of three groups of transcripts; two including genes related to growth, one including genes related to egg production. The function of the regulated transcripts is discussed in relation to post molting morphological changes in adult female salmon louse. There is clear evidence that transcription of the major yolk proteins is not induced before some of the post molting growth of abdomen and the genital segment has occurred. A hallmark for the observed growth is transcription of many putative cuticula proteins prior to the size increase.

Identification of differentially expressed ovarian genes during primary and early secondary oocyte growth in coho salmon, Oncorhynchus kisutch

BackgroundThe aim of this study was to identify differentially expressed ovarian genes during primary and early secondary oocyte growth in coho salmon, a semelparous teleost that exhibits synchronous follicle development.MethodsReciprocal suppression subtractive hybridization (SSH) libraries were generated from ovaries with perinucleolus (P) or cortical alveolus (CA) stage follicles and selected genes were assessed with quantitative PCR (qPCR). An assessment of changes in RNA composition during oocyte growth and its relationship to transcript levels was also conducted.ResultsSSH revealed several differentially expressed genes during early oogenesis, some which will not likely be utilized until 1–3 years later in salmon. Zona pellucida glycoprotein (zp) genes, vitellogenin receptor (vldlr) isoforms, cathepsin B (ctsba), cyclin E (ccne), a DnaJ transcript (dnaja2), and a ferritin subunit (fth3) were significantly elevated at the P stage, while a C-type lectin, retinol dehydrogenase (rdh1), and a coatomer protein subunit (cope) were upregulated at the CA stage. Putative follicle cell transcripts such as anti-Müllerian hormone (amh), lipoprotein lipase (lpl), apolipoprotein E (apoe), gonadal soma-derived growth factor (gsdf) and follicle-stimulating hormone receptor (fshr) also increased significantly at the CA stage. The analysis of RNA composition during oocyte growth showed that the total RNA yield and proportion of messenger RNA relative to non-polyadenylated RNAs declined as oogenesis progressed. This influenced apparent transcript levels depending on the type of RNA template used and normalization method.ConclusionIn coho salmon, which exhibit a dramatic change in oocyte size and RNA composition during oogenesis, use of messenger RNA as template and normalization of qPCR data to a housekeeping gene, ef1a, yielded results that best reflected transcript abundance within the ovarian follicle. Synthesis of zp transcripts and proteins involved in yolk incorporation and processing occurred during primary growth, while increased expression of a CA component and genes related to lipid incorporation occurred concomitant with the appearance of CA, but prior to lipid accumulation. Significant increases in transcripts for fshr, gsdf, and amh at the CA stage suggest a role of FSH and TGFβ peptides in previtellogenic oocyte growth and puberty onset in female salmon.

The combined effects of temperature and GnRHa treatment on the final stages of sexual maturation in Atlantic salmon (Salmo salar L.) females

Atlantic salmon (Salmo salar L.) females (2 SW), maturing for the first time, were reared under one of three temperature regimes (high: 14.3 +/- 0.5 degrees C; natural: 10.6 +/- 1.0 degrees C; and cold: 6.9 +/- 1.0 degrees C) in combination with one of two experimental treatments; an injection of GnRH analogue (GnRHa) contained in biodegradable microspheres, or a sham injection (microspheres only). The six experimental groups were then reared under simulated natural photoperiod for 4 weeks. Blood samples were drawn for analysis of plasma steroid levels and the fish were inspected for ovulation weekly. Batches of stripped eggs were incubated in triplicate incubators in raceways until the eyed stage. Treatment with GnRHa resulted in a substantial advancement and synchronization of ovulation at all temperatures, while exposure to cold water also appeared to advance ovulation slightly. While 75% (warm and cold) to 90% (natural) of GnRHa fish ovulated during the 4-week trial, only 30% of sham-treated females exposed to cold water, and none of the sham-treated fish held at higher temperatures, ovulated during this period. Survival rates of embryos to the eyed-stage were significantly higher for broodstock exposed to cold water. Plasma levels of testosterone (T), 17beta-oestradiol (E2), and 17alpha,20beta-dihydroxy-4-pregnen-3-one (17,20betaP) were all significantly affected by treatment with GnRHa and, to a lesser extent, temperature. The efficiency of GnRHa in counteracting the negative effects of high temperature on ovulation and the associated changes in circulating sex steroids suggest that temperature inhibition operates at least in part at the brain or pituitary.