Ecological Risk Assessment Paradigm for Salmon: Analyzing Immune Function to Evaluate Risk

Wild Pacific salmon populations are in serious decline, and as a result, a number of salmon stocks are listed as threatened or endangered under the Endangered Species Act. Our research identifies and supports the possibility that certain environmental contaminants can alter salmon survival, and as a result may contribute to these species being at risk. We have shown that juvenile chinook salmon (Oncorhynchus tshawytscha) are exposed to polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) as they migrate through a contaminated urban estuary in Puget Sound WA (the Duwamish Waterway estuary). Immune function was analyzed in these fish by examining the ability of their anterior kidney and splenic leukocytes to produce a primary and secondary in vitro plaque-forming cell (PFC) response to the hapten, trinitrophenyl (TNP), and by determining their susceptibility to a marine pathogen, Vibrio anguillarum. We found that fish outmigrating from the urban estuary produced a significantly lower PFC response to TNP and were more susceptible to the pathogen, compared to juvenile salmon collected from a rural estuary during their outmigration. In the laboratory, we exposed juvenile chinook salmon collected from a hatchery to either a PCB technical mixture or a PAH compound to determine if these contaminants have the potential to alter immune function in salmon. Indeed, we found that salmon exposed in the laboratory to either the PCB mixture or the PAH also produced lower PFC responses and were more susceptible to disease compared to animals treated with the solvent vehicle. In summary, contaminants such as PAHs and PCBs are demonstrated to influence salmon health, and thus have the potential to adversely impact salmon populations.     

Reproductive injury in English Sole (Pleuronectes vetulus) from the Hylebos Waterway, Commencement Bay, Washington

The effect of exposure to xenobiotic compounds on ovarian development was investigated in prespawning female English sole (Pleuronectes vetulus) from the Hylebos Waterway, an industrial site in Commencement Bay, WA, contaminated with aromatic hydrocarbons (AHs), polychlorinated biphenyls (PCBs)and other chlorinated compounds, including hexachlorobutadiene (HCBD) and hexachlorobenzene (HCB). Reference sole were collected from Colvos Passage, a nearby site with minimal sediment contaminant concentrations. English sole from theHylebos Waterway had significantly higher concentrations of fluorescent aromatic compounds (FACs) in bile, polycyclic aromatic compound-DNA adducts in liver, and dioxin-like and other selected PCB congeners in liver than sole from Colvos Passage. The Hylebos Waterway animals also showed significant alterations in their pattern of reproductive development when compared to Colvos Passage sole. Hylebos Waterway sole entered vitellogenesis at a nearlier age than Colvos Passage sole, with about 50%of fish below 5 years of age maturing in the Hylebos Waterway as compared to 20% of Colvos Passage sole in this age range, with corresponding increases in plasma estradiol concentrations and GSI in Hylebosfish. However, while the proportion of maturing Colvos Passage females increased with age to over70% for fish 5 years of age or greater, the proportion of maturing females in the Hylebos Waterway remained at about 50%. Moreover, plasma estradiol concentrations and gonadosomatic indices in these sole were depressed. Inhibited reproductive development and increased oocyte atresia in adult fish were correlated with elevated concentrations of FACs in bile. Enhanced growth, as well as exposure to both aromatic and chlorinated hydrocarbons, were associated with precocious maturation in sub adult Hylebos Waterway sole. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effects of turbidity and vegetation on the risk of juvenile salmonids, Oncorhynchus spp., to predation by adult cutthroat trout, O. clarkii

Synopsis During their seaward migration, juvenile salmonids encounter structural and visual cover which varies between and within watersheds. In this study, the effects of two types of cover (turbidity and artificial vegetation) on the predation mortality of juvenile salmonids exposed to fish piscivores was investigated in outdoor concrete ponds. During experiments, adult coastal cutthroat trout, Oncorhynchus clarkii clarkii, were allowed to feed on juvenile salmonid prey — chinook salmon, O. tshawytscha, chum salmon, O. keta, sockeye salmon, O. nerka, and cutthroat trout — in separate trials. Daily instantaneous per capita predation rate was determined for each turbidity and vegetation treatment, within each trial. Mean predation rates varied between 1% and 76% daily. In the presence of cover, mean daily predation rates were 10–75% lower than those in controls (no vegetation and clear water), depending on prey species. Predation rates were significantly lower in the presence of vegetation cover and did not covary with prey size or species. The effects of turbidity were generally not significant and were not additive with the effects of vegetation. However, turbidity appeared to significantly reduce the effectiveness of vegetation as cover for juvenile chinook and sockeye salmon. We suggest that these two forms of cover do not affect risk of predation by fish piscivores to juvenile salmonids via the same mechanism.     

Differential rejection of salmon lice by pink and chum salmon: disease consequences and expression of proinflammatory genes

The consequences of high (735 copepodids fish-1) and low (243 copepodids fish-1) level exposures of size-matched juvenile pink and chum salmon to Lepeophtheirus salmonis copepodids were examined. At both levels of exposure the prevalence and abundance of L. salmonis was significantly higher on chum salmon. In addition, the weight of exposed chum salmon following the high exposure was significantly less than that of unexposed chum salmon. At both exposures, the haematocrit of exposed chum salmon was significantly less than that of unexposed chum. Neither weight nor haematocrit of pink salmon was affected by exposures at these levels. Despite the presence of microscopic inflammatory lesions associated with attachment of L. salmonis on the epithelium of gill and fin of both salmon species, there were no mortalities following either exposure. A transient cortisol response was observed in chum salmon 21 d after low exposure. An earlier and quantitatively higher expression of the proinflammatory genes interleukin-8 (IL-8), tumour necrosis factor alpha-1 (TNFalpha-1) and interleukin-1beta (IL-1beta) in fin and head kidney of pink salmon suggested a mechanism of more rapid louse rejection in this species. Together, these observations indicate a relatively enhanced innate resistance to L. salmonis in the juvenile pink salmon compared with the juvenile chum salmon.     

Innate and enhanced predator recognition in hatchery-reared chinook salmon

We used a laboratory behaviour assay to investigate how innate predator recognition, handling stress, retention time, and number of conditioning events might affect chemically mediated anti-predator conditioning for hatchery-reared chinook salmon, Oncorhynchus tshawytscha. Juvenile chinook salmon with no prior exposure to predatory stimuli exhibited innate fright responses to northern pikeminnow, Ptychocheilis oregonensis, odour, regardless of whether the salmon came from a population that exists in sympatry or allopatry with northern pikeminnows. Juvenile chinook salmon exhibited enhanced predator recognition following a single conditioning event with conspecific extract and northern pikeminnow odour. Handling similar to what hatchery salmon might experience prior to release did not substantially reduce the conditioned response. When we conditioned juvenile chinook salmon in hatchery rearing vessels, fish from tanks treated once exhibited a conditioned response to northern pikeminnow odour in aquaria, but only for one behaviour (feeding response), and fish treated twice did not respond. The results suggest that enhanced recognition of predator stimuli occurs quickly, but may be to some extent context-specific, which may limit conditioned fright responses after release into the natural environment.     

Seawater adaptability of underyearling coho,chinook and chum salmons in the estuary of Avacha River (Kamchatka)

Synopsis The 96 h LC50 tests were conducted with chum, coho and chinook salmon alevins caught in the Avacha River (Kamchatka) estuary. Coho and chinook alevins exhibited very low seawater a adaptability when compared with chum. It is likely that coho and chinook young were pulled downstream by high velocity flow during the summer freshets.     

Vulnerability to predation and physiological stress responses of experimentally descaled juvenile chinook salmon,Oncorhynchus tshawytscha

Synopsis Juvenile salmonids,Oncorhynchus spp., commonly encounter conditions (e.g., during hatchery release and dam passage) that result in damage to the skin, scale, and slime complex. We conducted laboratory experiments to determine if descaling of juvenile chinook salmon,O. tshawytscha, increased their vulnerability to predation, and to assess the physiological stress responses elicited by descaling. Salmon were experimentally descaled on either 10% or 20% of their total body area. When offered equal numbers of control and descaled juvenile chinook salmon, northern squawfish,Ptychocheilus oregonensis, did not consume significantly more of either prey type (48–60% of consumed prey were descaled). Juvenile chinook salmon descaled on 10% of their body area did show significant physiological stress responses, however. Mean concentrations of plasma cortisol peaked 1 h after descaling, and returned to control levels by 12 h. Plasma glucose peaked 3 h post-treatment and remained elevated for 24 h. Plasma lactate increased immediately following treatment and returned to undisturbed control levels by 3 h. The osmoregulatory response of plasma potassium was highly variable, but plasma sodium decreased immediately and remained low for 24 h. The observed physiological responses suggest that descaling of juvenile chinook salmon could result in decreased resistance to disease and other stressors encountered in the field, possibly leading to reduced performance capacity and lowered survival.     

Juvenile chum salmon consumption of zooplankton in marine waters of southeastern Alaska: a bioenergetics approach to implications of hatchery stock interactions

Bioenergetics modeling was used to estimate zooplankton prey consumption of hatchery and unmarked stocks of juvenile chum salmon (Oncorhynchus keta) migrating seaward in littoral (nearshore) and neritic (epipelagic offshore) marine habitats of southeastern Alaska. A series of model runs were completed using biophysical data collected in Icy Strait, a regional salmon migration corridor, in May, June, July, August, and September of 2001. These data included a temperature (1-m surface versus surface to 20-m average), zooplankton standing crop (surface to 20-m depth versus entire water column), chum salmon diet (percent weight of prey type consumed), energy densities, and weight. Known numbers of hatchery releases were used in a cohort reconstruction model to estimate total abundance of hatchery and wild chum salmon in the northern region of southeastern Alaska, given average survival to adults and for two different (low and high) early marine littoral mortality rate assumptions. Total prey consumption was relatively insensitive to temperature differences associated with the depths potentially utilized by juvenile chum salmon. However, the magnitudes and temporal patterns of total prey consumed differed dramatically between the low and high mortality rate assumptions. Daily consumption rates from the bioenergetics model and CPUE abundance from sampling in Icy Strait were used to estimate amount and percentage of zooplankton standing crop consumed by mixed stocks of chum salmon. We estimated that only a small percentage of the available zooplankton was consumed by juvenile chum salmon, even during peak abundances of marked hatchery and unmarked mixed stocks in July. Total daily consumption of zooplankton by all stock groups of juvenile chum salmon was estimated to be between 330 and 1764 g/km²d¹ from June to September in the neritic habitat of Icy Strait. As with any modeling exercise, model outputs can be misleading if input parameters and underlying assumptions are not valid; therefore, additional studies are warranted, especially to determine physiological input parameters, and to improve abundance and mortality estimates specific to juvenile chum salmon. Future bioenergetics modeling is also needed to evaluate consumption by the highly abundant, vertically migrating planktivorous that co-occurred in our study; we suggest that these fishes have a greater impact on the zooplankton standing crop in Icy Strait than do hatchery stock groups of juvenile chum salmon.     

Density‐dependent mortality in Pacific salmon: the ghost of impacts past?

Conservation biologists often ignore density dependence because at‐risk populations are typically small relative to historical levels. However, if populations are reduced as a result of impacts that lower carrying capacity, then density‐dependent mortality may exist at low population abundances. Here, we explore this issue in threatened populations of juvenile chinook salmon (Oncorhynchus tshawytscha). We followed the fate of more than 50 000 juvenile chinook in the Snake River Basin, USA to test the hypothesis that their survival was inversely associated with juvenile density. We also tested the hypotheses that non‐indigenous brook trout and habitat quality affect the presence or strength of density dependence. Our results indicate that juvenile chinook suffer density‐dependent mortality and the strength of density dependence was greater in streams in which brook trout were absent. We were unable to detect an effect of habitat quality on the strength of density dependence. Historical impacts of humans have greatly reduced population sizes of salmon, and the density dependence we report may stem from a shortage of nutrients normally derived from decomposing salmon carcasses. Cohorts of juvenile salmon may experience density‐dependent mortality at population sizes far below historical levels and recovery of imperiled populations may be much slower than currently expected.     

Ontogenetic variations in the type and size of prey consumed by juvenile coho,Oncorhynchus kisutch,and chinook,O. tshawytscha,salmon

Synopsis Stomach contents of juvenile coho,Oncorhynchus kisutch, and chinook,O. tshawytscha, salmon collected in purse seines off the coast of Washington and Oregon were examined for variations related to predator size. There was a general trend toward increasing consumption of fish with increasing body size, due mainly to the increase in northern anchovy biomass consumed by the larger salmon. Most of the major prey taxa showed significant differences among the size classes examined for both salmon species. There was a direct relationship between predator and prey size for both coho and chinook, but considerable variation was found in prey length consumed within each size class. Prey width did not provide as good a fit as prey length for either species. In general, coho consumed larger fish prey in relation to their body length than chinook but there were substantial differences by month or year of collection.     

Wild chinook salmon survive better than hatchery salmon in a period of poor production

The population dynamics of chinook salmon (Oncorhynchus tshawytscha) from the Cowichan River on Vancouver Island, British Columbia, Canada are used by the Pacific Salmon Commission as an index of the general state of chinook salmon coast wide. In recent years the production declined to very low levels despite the use of a hatchery that was intended to increase production by improving the number of smolts entering the ocean. In 2008, we carried out an extensive study of the early marine survival of the hatchery and wild juvenile chinook salmon. We found that both rearing types mostly remained within the Gulf Islands study area during the period when most of the marine mortality occurred for the hatchery fish. By mid September, approximately 1.3% of all hatchery fish survived, compared to 7.8%–31.5% for wild fish. This six to 24 times difference in survival could negate an estimated increased egg-to-smolt survival of about 13% that is theorized to result through the use of a hatchery. Estimates of the early marine survival are approximate, but sufficient to show a dramatic difference in the response of the two rearing types to the marine nursery area. If the declining trend in production continues for both rearing types, modifications to the hatchery program are needed to improve survival or an emphasis on improving the abundances of wild stocks is necessary, or both. The discovery that the juvenile Cowichan River chinook salmon remain within a relatively confined area of the Gulf Islands within the Strait of Georgia offers an excellent opportunity to research the mechanisms that cause the early marine mortalities and hopefully contribute to a management that improves the production.     

Lack of trophic competition among wild and hatchery juvenile chum salmon during early marine residence in Taku Inlet,Southeast Alaska

Early marine trophic interactions of wild and hatchery chum salmon (Oncorhynchus keta) were examined as a potential cause for the decline in harvests of adult wild chum salmon in Taku Inlet, Southeast Alaska. In 2004 and 2005, outmigrating juvenile chum salmon were sampled in nearshore habitats of the inlet (spring) and in epipelagic habitat at Icy Strait (summer) as they approached the Gulf of Alaska. Fish were frozen for energy density determination or preserved for diet analyses, and hatchery stocks were identified from the presence of thermal marks on otoliths. We compared feeding intensity, diets, energy density, and size relationships of wild and hatchery stocks (n = 3123) across locations and weeks. Only hatchery fish feeding intensity was negatively correlated with fish abundance. In both years, hatchery chum salmon were initially larger and had greater energy density than wild fish, but lost condition in early weeks after release as they adapted to feeding on wild prey assemblages. Diets differed between the stocks at all inlet locations, but did not differ for hatchery salmon between littoral and neritic habitats in the outer inlet, where the stocks overlapped most. Both diets and energy density converged by late June. Therefore, if density-dependent interactions affect wild chum salmon, these effects must be very rapid because survivors in Icy Strait showed few differences. Our study also demonstrates that hatchery release strategies used near Taku Inlet successfully promote early spatial segregation and prey partitioning, which reduce the probability of competition between wild and hatchery chum salmon stocks.     

Effect of differential gastric evacuation and multispecies prey items on estimates of daily energy intake in juvenile chinook salmon

Synopsis The caloric density of stomach contents in juvenile chinook salmon,Oncorhynchus tshawytscha, was not affected by gastric evacuation, suggesting a constant caloric density of stomach contents during evacuation. Differences in the caloric density of prey consumed did affect caloric density of stomach contents over a 24-h period. Consumption of the amphipodCorophium sp. was associated with reduced caloric densities of stomach contents. During periods whenCorophium contributed more than 4% of the stomach contents, average caloric density declined from 5.56 to 5.33 kcal g^–1. Despite this difference, estimates of daily energy intake of juvenile chinook salmon were only 3%, greater when developed from the mean caloric density of stomach contents excludingCorophium.     

Incomplete congruence between morphobiological characters and sex-specific molecular markers in Pacific salmons: 1. Analysis of discrepancy in five species of the genus <Emphasis Type="Italic">Oncorhynchus</Emphasis>

The congruence between molecular markers, identifying the presence of the Y chromosome, and secondary sexual characters was examined in Asian populations of five Pacific salmon species: pink salmon (Oncorhynchus gorbuscha), chum salmon (O. keta), sockeye salmon (O. nerka), chinook salmon (O. tschawytscha), and sima (O. masou). It was demonstrated that in all species examined, the presence or absence of sex-specific molecular markers was to a considerable degree congruent with secondary sexual characters, but in some cases, an incongruence was found. These findings suggested that the mechanism underlying this phenomenon was similar or identical in all species examined. Possible genetic and physiological explanations of this phenomenon are discussed.     

Salmon growth hormone is transported into the circulation of rainbow trout,Oncorhynchus mykiss,after intestinal administration

Summary Transport of recombinant salmon growth hormone (rsGH) into the circulation of rainbow trout following intragastric or rectal administration was investigated. Changes in plasma GH levels were analyzed by radioimmunoassay specific to chum salmon GH. Intragastric administration of rsGH by oral intubation resulted in elevation of GH immunoreactivity in plasma after 11h. The plasma GH increased maximally after 15h, and then declined rapidly to reach a normal level after 19h. On the other hand, oral intubation of rsGH to carp, which has no stomach, caused elevation of plasma GH levels after 1 h which lasted for 21 h. In sharp contrast, rectal administration of rsGH to rainbow trout significantly elevated plasma GH levels after 15 min, which reached a maximum after 30 min. The rsGH was labeled with fluorescein isothiocyanate (FITC) to distinguish exogenous from endogenous GH, and administered into rainbow trout rectally. Subsequent analysis of plasma samples on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed two fluorescent bands at the same molecular weights as those of the monomeric and dimeric rsGHs. Intragastric and rectal administration of rsGH into juvenile trout resulted in significant increases in length and body weight compared to the control fish. This study strongly suggests that the rsGH is transported into the circulation of salmonids via the lower part of the intestine and takes part in growth stimulation.Abbreviations DCA deoxycholic acid - FITC fluorescein isothiocyanate - GH growth hormone - HPLC high-performance liquid chromatography - HRP horseradish peroxidase - RIA radioimmunoassay - rsGH recombinant salmon growth hormone - sGH chum salmon growth hormone - SEM standard error of mean - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - TFA trifluoroacetic acid     

Effect of hypo-osmotic environmental changes on the expression of gonadotropin-releasing hormone,its receptor,and gonadotropin hormone subunit mRNA in adult chum salmon (Oncorhynchus keta)

Migrating fish such as salmonids are affected by external environmental factors and salinity changes are particularly important, influencing spawning migration. The aim of this study was to test whether changes in salinity would affect the expression of the hypothalamic-pituitary-gonadal (HPG) axis hormones (gonadotropin-releasing hormones (GnRHs) [salmon GnRH and chicken GnRH-II], GnRH receptors [GnRHR1 and GnRHR5], and mRNA of the gonadotropin hormone [GTH] subunits [GTHα, follicle stimulating hormone β, and luteinizing hormone β]) in chum salmon (Oncorhynchus keta). Fish were progressively transferred from seawater (SW) through 50% SW to freshwater (FW), and the relationship between the osmoregulatory hormone prolactin (PRL) and sexual maturation was determined. The expression and activity of HPG hormones and their receptors, and levels of estradiol-17β and PRL increased after fish were transferred to FW, demonstrating that changes in salinity stimulate the HPG axis and PRL production in migrating chum salmon. These findings reveal details about the role of the endocrine system in maintaining homeostasis and stimulating sexual maturation and reproduction in response to salinity changes in this species.     

Effects of rapid experimental temperature increases on acute physiological stress and behaviour of stream dwelling juvenile chinook salmon

We experimentally heated small streams in summer and investigated the short-term behavioural changes and physiological stress responses of juvenile chinook salmon (Oncorhynchus tshawytscha). We rapidly raised temperatures ∼1–4 °C for 1.5 h above ambient levels of ∼7–15 °C in groundwater fed tributary streams and ∼19–23 °C in side-channel streams. Juvenile chinook rearing in groundwater fed tributaries were generally unaffected behaviourally; however, we found that temperature increase caused fish in the tributary trials to be physiologically stressed (elevations in mean cortisol concentrations ranged from 116% to 253%). Side-channel trials caused some mortality of juvenile chinook and a stronger display of behaviours indicative of stress and avoidance such as erratic swimming, abnormal posture, and aggregative behaviour. Foraging rates increased over 56 times in response to heating in side-channel trials. Cortisol levels did not increase in side-channel trials, but rather showed a trend to levels below control values suggesting an impaired stress response possibly due to chronic stress. Our results may reflect conservative responses in terms of what we may find with other salmonid species since juvenile chinook have been described as the most tolerant of the Pacific salmon species to elevated temperatures.     

Cloning preprolactin cDNA from Pacific salmon in Escherichia coli]

Prolactin coding mRNA was shown to be a prevalent part of chum salmon (Oncorhynchus keta) pituitary poly(A)-RNA during the spawning period. Clone lambda gtPrk12 was selected from the pituitary cDNA library by means of hybridization with the prolactin probe, and a nucleotide sequence of the insertion was determined and compared to the prolactin coding sequences from rainbow trout and Pacific chinook salmon, which had been published earlier. The sequences compared exhibited a significant homology. The deduced amino acid sequence of the chum salmon prolactin differed from a sequence determined directly in a single position. The prolactin-coding sequence can be used for constructing the bacterial strain producing prolactin.     

Functional response of juvenile pink and chum salmon: effects of consumer size and two types of zooplankton prey

Feeding rate experiments were conducted for pink salmon Oncorhynchus gorbuscha fry [mean fork length ( L _F) 39 mm], juveniles (103–104 mm L _F) and juvenile chum salmon Oncorhynchus keta (106–107 mm L _F). Fishes were presented with small copepod ( Tisbi sp.) or larger mysid shrimp ( Mysidopsis bahia ) prey at varying densities ranging from 1 to 235 prey l⁻¹ in feeding rate experiments conducted at water temperatures ranging from 10·5 to 12·0° C under high light levels and low turbidity conditions. Juvenile pink and chum salmon demonstrated a type II functional response to mysid and copepod prey. Mysid prey was readily selected by both species whereas the smaller bodied copepod prey was not. When offered copepods, pink salmon fry fed at a higher maximum consumption rate (2·5 copepods min⁻¹) than larger juvenile pink salmon (0·4 copepods min⁻¹), whereas larger juvenile chum salmon exhibited the highest feeding rate (3·8 copepods min⁻¹). When feeding on mysids, the maximum feeding rate for larger juvenile pink (12·3 mysids min⁻¹) and chum (11·5 mysids min⁻¹) salmon were similar in magnitude, and higher than feeding rates on copepods. Functional response models parameterized for specific sizes of juvenile salmon and zooplankton prey provide an important tool for linking feeding rates to ambient foraging conditions in marine environments, and can enable mechanistic predictions for how feeding and growth should respond to spatial-temporal variability in biological and physical conditions during early marine life stages.     

Experimental and natural host specificity of Loma salmonae (Microsporidia)

The microsporidian Loma salmonae (Putz, Hoffman & Dunbar, 1965) Morrison & Sprague, 1981 has caused significant gill disease in Pacific salmon Oncorhynchus spp. Host specificity of the parasite was examined experimentally by per os challenge of selected salmonids and non-salmonids with infective chinook salmon O. tshawytscha gill material. Pink Oncorhynchus gorbuscha and chum salmon O. keta, brown Salmo trutta and brook trout Salvelinus fontinalis, and chinook salmon (controls) were positive, whereas Atlantic salmon Salmo salar and Arctic char Salvelinus alpinus were negative. In addition, no non-salmonids were susceptible to experimental exposure. Wild Pacific salmon species in British Columbia, Canada, were examined for L. salmonae during their freshwater life history stages (smolts, prespawning, spawning). All stages were infected, although infections in smolts were only detectable using a L. salmonae-specific PCR test. Many previous Loma spp. described from Oncorhychus spp. are likely L. salmonae based on host, parasite morphology, and site of infection.