Intestinal lesions and parasites associated with senescence and prespawn mortality in Chinook Salmon (Oncorhynchus tshawytscha).

Prespawn mortality (PSM) presents a major problem for the recovery of spring Chinook Salmon (Oncorhynchus tshawytscha) populations. In the Willamette River, Oregon, PSM exceeds 90% in some years but factors explaining it are not well understood. We examined intestinal tissue samples using histological slides from over 783 spring Chinook Salmon collected between 2009 and 2021, which included tissues from PSM fish, artificially spawned captive broodstock (BS) and normal river run fish, comprised of trapped (Live) and naturally post-spawned river (RPS) fish collected from the river. We observed degeneration of the intestinal epithelium and loss of villous structure, with concurrent severe enteritis. A natural progression of decline in epithelial integrity (EI) through the summer and fall until spawning and subsequent death was also observed. Live fish exhibited high EI scores (mean = 68%), BS exhibited variable EI scores (35%) and RPS exhibited severe loss of EI (14%). PSM fish exhibited prominent loss of intestinal epithelium with EI scores (13%), very similar to RPS fish, despite having been collected earlier in the year. Hence, we argue that low EI scores are strongly linked with PSM. Ceratonova shasta and Enterocytozoon schreckii were common in all groups, but neither were linked to either PSM or a decline in EI.

Aeromonas salmonicida, causative agent of salmonid furunculosis, isolated from the freshwater parasitic copepod, Salmincola californiensis.

Here, we provide evidence that the freshwater parasitic copepod, Salmincola californiensis, acts as a vector for Aeromonas salmonicida. While investigating the effects of S. californiensis on Chinoook salmon (Oncorhynchus tshawytscha), we tangentially observed that fish infected with the copepod developed furunculosis, caused by A. salmonicida. This occurred despite being reared in pathogen-free well water in a research facility with no prior history of spontaneous infection. We further investigated the possibility of S. californiensis to serve as a vector for the bacterium via detection of fluorescently labelled A. salmonicida inside the egg sacs from copepods in which the fish hosts were experimentally infected with GFP-A449 A. salmonicida. We then evaluated copepod egg sacs that were collected from adult Chinook salmon from a freshwater hatchery with A. salmonicida infections confirmed by either culture or PCR. The bacterium was cultured on tryptic soy agar plates from 75% of the egg sacs, and 61% were positive by PCR. These three separate experiments indicate an alternative tactic of transmission in addition to direct transmission of A. salmonicida in captivity. The copepod may play an important role in transmission of the bacterium when fish are more dispersed, such as in the wild.

Gut microbiome composition associates with corticosteroid treatment, morbidity, and senescence in Chinook salmon (Oncorhynchus tshawytscha).

Pacific salmon experience prolonged elevation in corticosteroid hormones during important life history events including migration, reproduction, and senescence. These periods of elevated corticosteroids correspond with changes to immunity and energy metabolism; therefore, fish may be particularly vulnerable to mortality at these times. Recent studies found that stress-induced cortisol release associated with microbial community shifts in salmonids, raising the question of how longer-term corticosteroid dynamics that accompany life history transitions affect salmonid microbiomes. In this work, we experimentally evaluated the relationships between gut microbiome composition, chronically elevated corticosteroids, and mortality in juvenile Chinook salmon (Oncorhynchus tshawytscha). We found that treatment with slow-release implants of the corticosteroids cortisol or dexamethasone resulted in changes to the gut microbiome. Morbidity was also associated with microbiome composition, suggesting that the gut microbiome reflects individual differences in susceptibility to opportunistic pathogens. Additionally, we analyzed a small number of samples from adult fish at various stages of senescence. Results from these samples suggest that microbiome composition associated with gut integrity, and that the microbial communities of corticosteroid treated juveniles shift in composition toward those of senescent adults. Overall, findings from this work indicate that the gut microbiome correlates with mortality risk during periods of chronic corticosteroid elevation.

An assessment of terminology for intraspecific diversity in fishes, with a focus on "ecotypes" and "life histories".

Understanding and preserving intraspecific diversity (ISD) is important for species conservation. However, ISD units do not have taxonomic standards and are not universally recognized. The terminology used to describe ISD is varied and often used ambiguously. We compared definitions of terms used to describe ISD with use in recent studies of three fish taxa: sticklebacks (Gasterosteidae), Pacific salmon and trout (Oncorhynchus spp., "PST"), and lampreys (Petromyzontiformes). Life history describes the phenotypic responses of organisms to environments and includes biological parameters that affect population growth or decline. Life-history pathway(s) are the result of different organismal routes of development that can result in different life histories. These terms can be used to describe recognizable life-history traits. Life history is generally used in organismal- and ecology-based journals. The terms paired species/species pairs have been used to describe two different phenotypes, whereas in some species and situations a continuum of phenotypes may be expressed. Our review revealed overlapping definitions for race and subspecies, and subspecies and ecotypes. Ecotypes are genotypic adaptations to particular environments, and this term is often used in genetic- and evolution-based journals. "Satellite species" is used for situations in which a parasitic lamprey yields two or more derived, nonparasitic lamprey species. Designatable Units, Evolutionary Significant Units (ESUs), and Distinct Population Segments (DPS) are used by some governments to classify ISD of vertebrate species within distinct and evolutionary significant criteria. In situations where the genetic or life-history components of ISD are not well understood, a conservative approach would be to call them phenotypes.

Laboratory infection rates and associated mortality of juvenile Chinook Salmon (Oncorhynchus tshawytscha) from parasitic copepod (Salmincola californiensis).

Pacific salmon (Oncorhynchus spp.) rearing in lakes and reservoirs above dams have been known to become heavily infected with an ectoparasitic copepod (Salmincola californiensis). Little is known about the factors that affect the parasite infection prevalence and intensity. However, previous research suggests that the parasite may negatively affect the fitness and survival of the host fish. The effect of water temperature, confinement and the density of the free-swimming infectious stage of S. californiensis, the copepodid, on infection prevalence and intensity was evaluated by experimentally exposing juvenile Chinook Salmon (O. tshawytscha). Infection rates observed in wild populations were achieved under warm water (15-16°C) and high copepodid density (150-300/L) treatment conditions. Infection prevalence and intensity were also significantly higher in larger fish. During the infection experiment, 4.5% of infected fish died within 54 days with mortality significantly related to copepod infection intensity. The potential for autoinfection was compared to cross-infection by cohabitation of infected fish with naïve fish. Previously infected fish had significantly greater infection intensity compared with naïve fish, indicating that infected fish can be reinfected and that they may be more susceptible than naïve fish.

Rearing environment affects spatial learning in juvenile Chinook salmon Oncorhynchus tshawytscha.

We tested the prediction that a complex physical rearing environment would enhance short-term spatial memory as assessed by learning ability in a spatial navigation task in juvenile Chinook salmon Oncorhynchus tshawytscha. We reared fish in two low-density treatments, where fish were either in bare fiberglass tanks (bare) or in tanks with physical structure (complex). We also tested conventionally reared high-density hatchery fish to compare with these other experimental treatments. Our reason for including this third hatchery treatment is that the two low-density treatments, aside from the manipulation of structure, followed a rearing programme that is designed to produce fish with more wild-like characteristics. We tested individually marked fish for seven consecutive days and recorded movement and time to exit a testing maze. Stimulus conspecific fish outside the exit of the maze provided positive reinforcement for test fish. Fish from the bare treatment were less likely to exit the start box compared with fish in the complex and hatchery treatments. However, fish in the hatchery treatment were significantly more likely to exit the maze on their own compared with both the bare and complex treatments. Hatchery fish effectively learned the task as shown by a decrease in the number of mistakes over time, but the number of mistakes was significantly greater on the first day of trials. Increasing habitat complexity with structure may not necessarily promote spatial learning ability, but differences between hatchery and experimental treatments in rearing density and motivation to be near conspecifics likely led to observed behavioural differences.

Egg size and growth in steelhead Oncorhynchus mykiss.

The effects of egg size on early development and growth of steelhead Oncorhynchus mykiss were recorded for more than 200 days following hatching. Fish from smaller eggs hatched sooner and at a smaller size than fish from larger eggs, but fish from smaller eggs showed consistently higher growth rates than fish from larger eggs. Since many life-history attributes appear to be determined by size or growth rate at age during the first year, egg size could be a significant predictor of important changes in the life history of individuals.

The effect of rearing structures on behaviour and movement of juvenile steelhead Oncorhynchus mykiss.

The relative movement of juvenile steelhead Oncorhynchus mykiss reared on two treatments was investigated to provide insight on the effect of structure in the rearing environment on the behaviour of the fish before potential release into a natural river system. The progeny of wild broodstock were reared either in the presence or absence of structure in the tank environment for 7 months at the Oregon Hatchery Research Center located in Oregon, U.S.A. Behavioural assessments, including movement response to a simulated predator, showed that fish reared on structure moved a similar amount (based on line crosses) as fish reared in bare hatchery tanks. No significant difference was observed in the proportion of time spent near a small structure within the behavioural test tank between the two treatments, but all fish showed decreased movement over time with each subsequent predation event. Fish from both treatments spent 30% of their time in the section of the tank containing the structure, which was one section out of a total of eight sections. In both treatments, fish foraged 20% of the time in the 2 min following the introduction of live tubifex prey (two separate events). Overall, similar movement and foraging responses occurred following mock predation events for juveniles reared either with or without structure. Developing assessment tools such as these, that measures behaviours related to survival based on rearing environment should allow managers to better predict the survival and effect of rearing conditions on the release of hatchery-origin fish into the wild.

Egg size and emergence timing affect morphology and behavior in juvenile Chinook Salmon, Oncorhynchus tshawytscha.

Variation in early life history traits often leads to differentially expressed morphological and behavioral phenotypes. We investigated whether variation in egg size and emergence timing influence subsequent morphology associated with migration timing in juvenile spring Chinook Salmon, Oncorhynchus tshawytscha. Based on evidence for a positive relationship between growth rate and migration timing, we predicted that fish from small eggs and fish that emerged earlier would have similar morphology to fall migrants, while fish from large eggs and individuals that emerged later would be more similar to older spring yearling migrants. We sorted eyed embryos within females into two size categories: small and large. We collected early and late-emerging juveniles from each egg size category. We used landmark-based geometric morphometrics and found that egg size appears to drive morphological differences. Egg size shows evidence for an absolute rather than relative effect on body morphology. Fish from small eggs were morphologically more similar to fall migrants, while fish from large eggs were morphologically more similar to older spring yearling migrants. Previous research has shown that the body morphology of fish that prefer the surface or bottom location in a tank soon after emergence also correlates with the morphological variations between wild fall and spring migrants, respectively. We found that late-emerging fish spent more time near the surface. Our study shows that subtle differences in early life history characteristics may correlate with a diversity of future phenotypes.

Pacific lamprey (Entosphenus tridentatus) ammocoetes exposed to contaminated Portland Harbor sediments: Method development and effects on survival, growth, and behavior.

Many anthropogenic disturbances have contributed to the decline of Pacific lampreys (Entosphenus tridentatus), but potential negative effects of contaminants on lampreys are unclear. Lamprey ammocoetes are the only detritivorous fish in the lower Willamette River, Oregon, USA, and have been observed in Portland Harbor sediments. Their long benthic larval stage places them at risk from the effects of contaminated sediment. The authors developed experimental methods to assess the effects of contaminated sediment on the growth and behavior of field-collected ammocoetes reared in a laboratory. Specifically, they developed methods to assess individual growth and burrowing behavior. Burrowing performance demonstrated high variability among contaminated sediments; however, ammocoetes presented with noncontaminated reference sediment initiated burrowing more rapidly and completed it faster. Ammocoete reemergence from contaminated sediments suggests avoidance of some chemical compounds. The authors conducted long-term exposure experiments on individually held ammocoetes using sediment collected from their native Siletz River, which included the following: contaminated sediments collected from 9 sites within Portland Harbor, 2 uncontaminated reference sediments collected upstream, 1 uncontaminated sediment with characteristics similar to Portland Harbor sediments, and clean sand. They determined that a 24-h depuration period was sufficient to evaluate weight changes and observed no mortality or growth effects in fish exposed to any of the contaminated sediments. However, the effect on burrowing behavior appeared to be a sensitive endpoint, with potentially significant implications for predator avoidance. Environ Toxicol Chem 2016;35:2092-2102. © 2016 SETAC.

Innate and adaptive immune responses in migrating spring-run adult chinook salmon, Oncorhynchus tshawytscha.

Adult Chinook salmon (Oncorhynchus tshawytscha) migrate from salt water to freshwater streams to spawn. Immune responses in migrating adult salmon are thought to diminish in the run up to spawning, though the exact mechanisms for diminished immune responses remain unknown. Here we examine both adaptive and innate immune responses as well as pathogen burdens in migrating adult Chinook salmon in the Upper Willamette River basin. Messenger RNA transcripts encoding antibody heavy chain molecules slightly diminish as a function of time, but are still present even after fish have successfully spawned. In contrast, the innate anti-bacterial effector proteins present in fish plasma rapidly decrease as spawning approaches. Fish also were examined for the presence and severity of eight different pathogens in different organs. While pathogen burden tended to increase during the migration, no specific pathogen signature was associated with diminished immune responses. Transcript levels of the immunosuppressive cytokines IL-10 and TGF beta were measured and did not change during the migration. These results suggest that loss of immune functions in adult migrating salmon are not due to pathogen infection or cytokine-mediated immune suppression, but is rather part of the life history of Chinook salmon likely induced by diminished energy reserves or hormonal changes which accompany spawning.

Salinity effects on plasma ion levels, cortisol, and osmolality in Chinook salmon following lethal sampling.

Studies on hydromineral balance in fishes frequently employ measurements of electrolytes following euthanasia. We tested the effects of fresh- or salt-water euthanasia baths of tricaine mesylate (MS-222) on plasma magnesium (Mg(2+)) and sodium (Na(+)) ions, cortisol and osmolality in fish exposed to saltwater challenges, and the ion and steroid hormone fluctuations over time following euthanasia in juvenile spring Chinook salmon (Oncorhynchus tshawytscha). Salinity of the euthanasia bath affected plasma Mg(2+) and Na(+) concentrations as well as osmolality, with higher concentrations in fish euthanized in saltwater. Time spent in the bath positively affected plasma Mg(2+) and osmolality, negatively affected cortisol, and had no effect on Na(+) concentrations. The difference of temporal trends in plasma Mg(2+) and Na(+) suggests that Mg(2+) may be more sensitive to physiological changes and responds more rapidly than Na(+). When electrolytes and cortisol are measured as endpoints after euthanasia, care needs to be taken relative to time after death and the salinity of the euthanasia bath.

Designing a Monitoring Program to Estimate Estuarine Survival of Anadromous Salmon Smolts: Simulating the Effect of Sample Design on Inference.

A number of researchers have attempted to estimate salmonid smolt survival during outmigration through an estuary. However, it is currently unclear how the design of such studies influences the accuracy and precision of survival estimates. In this simulation study we consider four patterns of smolt survival probability in the estuary, and test the performance of several different sampling strategies for estimating estuarine survival assuming perfect detection. The four survival probability patterns each incorporate a systematic component (constant, linearly increasing, increasing and then decreasing, and two pulses) and a random component to reflect daily fluctuations in survival probability. Generally, spreading sampling effort (tagging) across the season resulted in more accurate estimates of survival. All sampling designs in this simulation tended to under-estimate the variation in the survival estimates because seasonal and daily variation in survival probability are not incorporated in the estimation procedure. This under-estimation results in poorer performance of estimates from larger samples. Thus, tagging more fish may not result in better estimates of survival if important components of variation are not accounted for. The results of our simulation incorporate survival probabilities and run distribution data from previous studies to help illustrate the tradeoffs among sampling strategies in terms of the number of tags needed and distribution of tagging effort. This information will assist researchers in developing improved monitoring programs and encourage discussion regarding issues that should be addressed prior to implementation of any telemetry-based monitoring plan. We believe implementation of an effective estuary survival monitoring program will strengthen the robustness of life cycle models used in recovery plans by providing missing data on where and how much mortality occurs in the riverine and estuarine portions of smolt migration. These data could result in better informed management decisions and assist in guidance for more effective estuarine restoration projects.

Occupancy modeling for improved accuracy and understanding of pathogen prevalence and dynamics.

Most pathogen detection tests are imperfect, with a sensitivity < 100%, thereby resulting in the potential for a false negative, where a pathogen is present but not detected. False negatives in a sample inflate the number of non-detections, negatively biasing estimates of pathogen prevalence. Histological examination of tissues as a diagnostic test can be advantageous as multiple pathogens can be examined and providing important information on associated pathological changes to the host. However, it is usually less sensitive than molecular or microbiological tests for specific pathogens. Our study objectives were to 1) develop a hierarchical occupancy model to examine pathogen prevalence in spring Chinook salmon Oncorhynchus tshawytscha and their distribution among host tissues 2) use the model to estimate pathogen-specific test sensitivities and infection rates, and 3) illustrate the effect of using replicate within host sampling on sample sizes required to detect a pathogen. We examined histological sections of replicate tissue samples from spring Chinook salmon O. tshawytscha collected after spawning for common pathogens seen in this population: Apophallus/echinostome metacercariae, Parvicapsula minibicornis, Nanophyetus salmincola/ metacercariae, and Renibacterium salmoninarum. A hierarchical occupancy model was developed to estimate pathogen and tissue-specific test sensitivities and unbiased estimation of host- and organ-level infection rates. Model estimated sensitivities and host- and organ-level infections rates varied among pathogens and model estimated infection rate was higher than prevalence unadjusted for test sensitivity, confirming that prevalence unadjusted for test sensitivity was negatively biased. The modeling approach provided an analytical approach for using hierarchically structured pathogen detection data from lower sensitivity diagnostic tests, such as histology, to obtain unbiased pathogen prevalence estimates with associated uncertainties. Accounting for test sensitivity using within host replicate samples also required fewer individual fish to be sampled. This approach is useful for evaluating pathogen or microbe community dynamics when test sensitivity is <100%.

Migratory Patterns of Wild Chinook Salmon Oncorhynchus tshawytscha Returning to a Large, Free-Flowing River Basin.

Upriver movements were determined for Chinook salmon Oncorhynchus tshawytscha returning to the Yukon River, a large, virtually pristine river basin. These returns have declined dramatically since the late 1990s, and information is needed to better manage the run and facilitate conservation efforts. A total of 2,860 fish were radio tagged during 2002-2004. Most (97.5%) of the fish tracked upriver to spawning areas displayed continual upriver movements and strong fidelity to the terminal tributaries entered. Movement rates were substantially slower for fish spawning in lower river tributaries (28-40 km d-1) compared to upper basin stocks (52-62 km d-1). Three distinct migratory patterns were observed, including a gradual decline, pronounced decline, and substantial increase in movement rate as the fish moved upriver. Stocks destined for the same region exhibited similar migratory patterns. Individual fish within a stock showed substantial variation, but tended to reflect the regional pattern. Differences between consistently faster and slower fish explained 74% of the within-stock variation, whereas relative shifts in sequential movement rates between "hares" (faster fish becoming slower) and "tortoises" (slow but steady fish) explained 22% of the variation. Pulses of fish moving upriver were not cohesive. Fish tagged over a 4-day period took 16 days to pass a site 872 km upriver. Movement rates were substantially faster and the percentage of atypical movements considerably less than reported in more southerly drainages, but may reflect the pristine conditions within the Yukon River, wild origins of the fish, and discrete run timing of the returns. Movement data can provide numerous insights into the status and management of salmon returns, particularly in large river drainages with widely scattered fisheries where management actions in the lower river potentially impact harvests and escapement farther upstream. However, the substantial variation exhibited among individual fish within a stock can complicate these efforts.

Effect of short-term decrease in water temperature on body temperature and involvement of testosterone in steelhead and rainbow trout, Oncorhynchus mykiss.

The Pacific salmonid species Oncorhynchus mykiss is separated into a migratory form (steelhead trout) and a non-migratory form (rainbow trout). A decrease in water temperature is likely a cue triggering downstream behavior in the migratory form, and testosterone inhibits onset of this behavior. To elucidate differences in sensitivity to water temperature decreases between the migratory and non-migratory forms and effect of testosterone on the sensitivity, we examined two experiments. In experiment 1, we compared changes in body temperature during a short-term decrease in water temperature between both live and dead steelhead and rainbow trout. In experiment 2, we investigated effects of testosterone on body temperature decrease in steelhead trout. Water temperature was decreased by 3°C in 30min. The body temperature of the steelhead decreased faster than that of the rainbow trout. In contrast, there was no significant difference in the decrease in body temperature between dead steelhead and rainbow trout specimens. The body temperature of the testosterone-treated steelhead trout decreased more slowly than that of control fish. Our results suggest that the migratory form is more sensitive to decreases in water temperature than the non-migratory form. Moreover, testosterone might play an inhibitory role in sensitivity to such decreases.

Sensitivity and specificity of histology for diagnoses of four common pathogens and detection of nontarget pathogens in adult Chinook salmon (Oncorhynchus tshawytscha) in fresh water.

Histology is often underutilized in aquatic animal disease screening and diagnostics. The agreement between histological classifications of infection and results using diagnostic testing from the American Fisheries Society's Blue Book was conducted with 4 common salmon pathogens: Aeromonas salmonicida, Renibacterium salmoninarum, Ceratomyxa shasta, and Nanophyetus salmincola. Adult Chinook salmon (Oncorhynchus tshawytscha) in Oregon were evaluated, and agreement between tests was calculated. Live and dead (both pre- and postspawning) salmon were collected from the Willamette River, Oregon, its tributaries, the Willamette Hatchery, and after holding in cool, pathogen-free water during maturation at Oregon State University. Sensitivity and specificity of histology compared to Blue Book methods for all fish, live fish only, and dead (pre- and postspawned combined) fish only were, respectively, as follows: A. salmonicida (n = 105): specificity 87.5%, 87.5%, 87.5% and sensitivity 38.6%, 14.8%, 60.0%; R. salmoninarum (n = 111): specificity 91.9%, 85.7%, 97.7% and sensitivity 16.0%, 7.1%, 27.2%; C. shasta (n = 136): specificity 56.0%, 63.3%, 28.6% and sensitivity 83.3%, 86.2%, 71.4%; N. salmincola (n = 228): specificity 68.2%, 66.7%, not possible to calculate for dead fish and sensitivity 83.5%, 80.5%, 87.3%. The specificity was good for bacterial pathogens. This was not the case for C. shasta, likely due to detection of presporogenic forms only by histology. Sensitivity of histology for bacterial pathogens was low with the exception of dead fish with A. salmonicida. Kappa analysis for agreement between Blue Book and histology methods was poor to moderate. However, histological observations revealed the presence of other pathogens that would not be detected by other methods.

Effect of commercially available egg cures on the survival of juvenile salmonids.

There is some concern that incidental consumption of eggs cured with commercially available cures for the purpose of sport fishing causes mortality in juvenile salmon. We evaluated this by feeding juvenile spring Chinook (Oncorhynchus tshawytscha) and steelhead (O. mykiss) with eggs cured with one of five commercially available cures. We observed significant levels of mortality in both pre-smolts and smolts. Depending on the experiment, 2, 3, or 4 of the cures were associated with mortality. Mortality tended to be higher in the smolts than in the parr, but there was no clear species effect. The majority of mortality occurred within the first 10 d of feeding. Removal of sodium sulfite from the cure significantly reduced the level of mortality. Soaking the eggs prior to feeding did not reduce mortality. We observed a clear relationship between the amount of cured egg consumed each day and the survival time. We conclude that consumption of eggs cured with sodium sulfite has the potential to cause mortality in juvenile steelhead and Chinook salmon in the wild.

Persistence of infection by metacercariae of Apophallus sp., Neascus sp., and Nanophyetus salmincola plus two myxozoans (Myxobolus insidiosus and Myxobolus fryeri) in coho salmon Oncorhynchus kisutch.

We evaluated the ability of 5 muscle- or skin-dwelling parasites to persist in naturally infected coho salmon, Oncorhynchus kisutch, from the West Fork Smith River, Oregon, by holding them in captivity from late summer to early spring (parr stage to the typical time of smoltification). These parasites included metacercariae of 3 digeneans, Nanophyetus salmincola, Apophallus sp., and neascus sp., and 2 myxozoans, Myxobolus insidiosus and Myxobolus fryeri. Two groups of wild-caught fish were evaluated in the laboratory, i.e., heavily infected fish from the lower main stem and less severely infected fish collected from tributaries of this river. All parasites survived in these fish for the 7-month experiment. Only 2 parasites had a statistically significant lower median abundance between host life stages. The mean abundance of N. salmincola declined 45% in the tributary fish and Apophallus sp. declined 43% in the lower main stem fish. However, more than 50% of each species persisted until the end of the study, with smolts still harboring relatively high infections.