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Carla M. Conway

Publications and source records attributed to Carla M. Conway.

At least 19 recordsLinked to original sources

Poor population recovery of Lake Washington Chinook Salmon (Oncorhynchus tshawytscha) in Pacific Northwestern USA may be associated with disease caused by Ceratonova shasta in an urban-influenced watershed

Understanding the effects of anthropogenic changes on fish disease is vital for supporting mitigative actions for fisheries population recovery. The Lake Washington watershed in the Pacific Northwestern United States is influenced by high urban development and climate change, compromising the sustainability of anadromous salmon. We investigated Ceratonova shasta , a myxozoan parasite, in Chinook Salmon Oncorhynchus tshawytscha to determine the impacts of C. shasta -induced enteronecrosis during salmon freshwater migrations within this watershed. All sampled adult Chinook Salmon returning to an enhancement hatchery to spawn were infected with genotype I C. shasta with a high prevalence of severe enteritis and enteronecrosis. Parasite loads and associated pathology were more severe in adult fish succumbing to prespawn mortality than in fish that spawned. Natural prespawn mortality of Chinook Salmon in the Cedar River was also associated with high C. shasta loads. Sentinel exposure studies with Chinook Salmon juveniles in the spring and fall demonstrated high levels of C. shasta genotype II actinospores in the Lake Washington Ship Canal, a human engineered canal that salmon must traverse in their freshwater migrations from the Puget Sound. The sentinel juvenile Chinook Salmon succumbed to clinical disease and high mortality from genotype II C. shasta , suggesting that these stocks have high susceptibility to this genotype. The results show that both genotypes I and II pose disease risks to adult and juvenile stages of Chinook Salmon in the watershed, and that the Lake Washington Ship Canal could be a C. shasta ‘hot spot’ for genotype II that may increase disease risk for juveniles during smolt emigration.

Washington

Host jump of an exotic fish rhabdovirus into a new class of animals poses a disease threat to amphibians

Spring viremia of carp virus (SVCV) is a rhabdovirus that primarily infects cyprinid finfishes and causes a disease notifiable to the World Organization for Animal Health. Amphibians, which are sympatric with cyprinids in freshwater ecosystems, are considered non-permissive hosts of rhabdoviruses. The potential host range expansion of SVCV in an atypical host species was evaluated by testing the susceptibility of amphibians native to the Pacific Northwest. Larval long-toed salamanders Ambystoma macrodactylum and Pacific tree frog Pseudacris regilla tadpoles were exposed to SVCV strains from genotypes Ia, Ib, Ic, or Id by either intraperitoneal injection, immersion, or cohabitation with virus-infected koi Cyprinus rubrofuscus . Cumulative mortality was 100% for salamanders injected with SVCV, 98–100% for tadpoles exposed to virus via immersion, and 0–100% for tadpoles cohabited with SVCV-infected koi. Many of the animals that died exhibited clinical signs of disease and SVCV RNA was found by in situ hybridization in tissue sections of immersion-exposed tadpoles, particularly in the cells of the gastrointestinal tract and liver. SVCV was also detected by plaque assay and RT-qPCR testing in both amphibian species regardless of the virus exposure method, and viable virus was detected up to 28 days after initial exposure. Recovery of infectious virus from naïve tadpoles cohabited with SVCV-infected koi further demonstrated that SVCV transmission can occur between classes of ectothermic vertebrates. Collectively, these results indicated that SVCV, a fish rhabdovirus, can be transmitted to and cause lethal disease in two amphibian species. Therefore, members of all five of the major vertebrate groups (mammals, birds, reptiles, fish, and amphibians) appear to be vulnerable to rhabdovirus infections. Future research studying potential spillover and spillback infections of aquatic rhabdoviruses between foreign and domestic amphibian and fish species will provide insights into the stressors driving novel interclass virus transmission events.

Viruses

Temporal, environmental, and demographic correlates of Ichthyophonus sp. infections in mature Pacific herring populations

Causes of population collapse and failed recovery often remain enigmatic in marine forage fish like Pacific herring ( Clupea pallasii ) that experience dramatic population oscillations. Diseases such as ichthyophoniasis are hypothesized to contribute to these declines, but lack of long-term datasets frequently prevents inference. Analysis of pathogen surveillance and population assessment datasets spanning 2007–2019 indicate that the age-based prevalence estimate of Ichthyophonus infection was, on average, 54% greater among a collapsed population of Pacific herring (Prince William Sound, Alaska, USA) as compared to a nearby population (Sitka Sound, Alaska, USA) that is relatively robust. During the study years, the age-based infection prevalence ranged from 14 to 44% in Prince William Sound and 5 to 33% in Sitka Sound. At both sites, the age-based infection prevalence declined over time, with an average decrease of 7% per year. Statistical analyses indicated that infection prevalence between the two populations was reduced by regional factors affecting both sites, and that these factors were independent of herring density. Infection prevalence in both populations was positively correlated with herring age and negatively correlated with the Pacific Decadal Oscillation. This study demonstrates how synthesis of environmental, stock assessment, and disease assessment data can be leveraged to elucidate epidemiological trends in diseases of wild fish.

ICES Journal of Marine Science

Exposure to 17α-ethinylestradiol results in differential susceptibility of largemouth bass (Micropterus salmoides) to bacterial infection

Disease outbreaks, skin lesions, mortality events, and reproductive abnormalities have been observed in wild populations of centrarchids. The presence of estrogenic endocrine disrupting compounds (EEDCs) has been implicated as a potential causal factor for these effects. The effects of prior EEDC exposure on immune response were examined in juvenile largemouth bass ( Micropterus salmoides ) exposed to a potent synthetic estrogen (17α-ethinylestradiol, EE2) at a low (EE2 Low , 0.87 ng/L) or high (EE2 High , 9.08 ng/L) dose for 4 weeks, followed by transfer to clean water and injection with an LD 40 dose of the Gram-negative bacteria Edwardsiella piscicida . Unexpectedly, this prior exposure to EE2 High significantly increased survivorship at 10 d post-infection compared to solvent control or EE2 Low -exposed, infected fish. Both prior exposure and infection with E. piscicida led to significantly reduced hepatic glycogen levels, indicating a stress response resulting in depletion of energy stores. Additionally, pathway analysis for liver and spleen indicated differentially expressed genes associated with immunometabolic processes in the mock-injected EE2 High treatment that could underlie the observed protective effect and metabolic shift in EE2 High -infected fish. Our results demonstrate that exposure to a model EEDC alters metabolism and immune function in a fish species that is ecologically and economically important in North America.

Environmental Science and Technology

Evaluating the effect of nuclear inclusion X (NIX) infections on Pacific razor clam populations

ABSTRACT: Nuclear inclusion X (NIX), the etiological agent of bacterial gill disease in Pacific razor clams Siliqua patula , was associated with host mortality events in coastal Washington State, USA, during the mid-1980s. Ongoing observations of truncated razor clam size distributions in Kalaloch Beach, Washington, raised concerns that NIX continues to impact populations. We conducted a series of spatial and longitudinal NIX surveillances, examined archived razor clam gill tissue, and used population estimates from stock assessments to test whether (1) the prevalence and intensity of NIX infections is higher at Kalaloch Beach relative to nearby beaches, (2) infected gill tissue has features consistent with historical descriptions of NIX-associated histopathology, and (3) annual clam survival is inversely related to NIX infection prevalence and intensity. NIX prevalence exceeded 85% at all sampled locations, and infection intensity was the highest at Kalaloch Beach by 0.9-2.6 orders of magnitude. Kalaloch Beach clams revealed histopathology consistent with previous NIX epidemics, including enlarged and/or rupturing branchial epithelial cells, branchial necrosis, and high hemocyte densities. Estimated annual survival was 22% at Kalaloch Beach, and ranged between 57 and 99% at other study sites. NIX infection intensity (via quantitative PCR) was not significantly correlated with annual survival; however, annual survival was lowest at Kalaloch Beach, where infection intensities were highest, suggesting that clams can tolerate infections up to a lethal threshold. Collectively these data support the hypothesis that high NIX intensities are associated with host mortality. NIX-associated mortality appears to be more pronounced at Kalaloch Beach relative to other Washington beaches.

Diseases of Aquatic Organisms

Survival and growth of suckers in mesocosms at three locations within Upper Klamath Lake, Oregon, 2018

Executive Summary Due to high mortality in the first year or two of life, Lost River ( Deltistes luxatus sp.) and Shortnose suckers ( Chasmistes brevirostris sp.) in Upper Klamath Lake, Oregon rarely reach maturity. In 2015, the U.S. Fish and Wildlife Service began the Sucker Assisted Rearing Program (SARP) to improve early life survival before releasing the fish back into Upper Klamath Lake. Survival and growth rates were compared for fish in mesocosms among three potential release or in-lake rearing sites, and in a pond at the SARP rearing facility. Fish used in this study included a mix of Lost River, Shortnose, and Klamath largescale suckers reared at either U.S. Fish and Wildlife Service or Klamath Tribes fish rearing facilities. These sites were Shoalwater Bay (SWB), Rattlesnake Point (RPT), and Cove Point (CPT). Ninety-nine to 103 suckers tagged with passive integrated transponders (PIT) were placed into each mesocosm for up to 80 days and up to 103 days in the SARP pond. Cessation of movement, as determined by passive detection of tagged fish on remote antennas, indicated mortality. Dissolved-oxygen saturation, temperature, and pH were tracked hourly in each mesocosm. All the suckers placed into the SWB mesocosm died during an extreme hypoxia event. These fish were replaced with another 120 PIT-tagged and 2 untagged hatchery-reared Lost River suckers from the Klamath Tribes Fish Research Facility (KTFRF), of which, all but two died during a second extreme hypoxia event. It was determined that SWB was an unsuitable site for summertime release or rearing of juvenile suckers in 2018. The summer survival rate was ≥86 percent at CPT, RPT, and the SARP pond. Suckers in the SARP pond grew slightly slower and gained less weight relative to increases in length than suckers held at RPT and CPT. All suckers sampled at the start of the study from both the SARP facility and the KTFRF, when water temperatures averaged approximately 18–22 degrees Celsius (°C), were infected with low levels of the gill parasite Ichthyobodo sp. Ichthyobodo sp. was detected on only 1 of 16 suckers sampled from CPT, RPT, and the SARP pond in late September or early October when water temperatures were approximately 16–19 °C, indicating fish were able to shed the parasite in cooler temperatures. Water quality conditions at RPT and CPT were adequate for in-lake rearing of SARP suckers in 2018. Due to interannual differences in water quality conditions, these sites may not be suitable in all years. Future research focused on the suitability of RPT, CPT and other potential sites under in years with varying conditions would be beneficial for improving sucker in-lake rearing practices. Additional research could help to elucidate how size at entry into the mesocosms affects sucker survival.

Oregon

Novel diagnostic tests for the putative agent of bacterial gill disease in Pacific razor clams (Siliqua patula)

Nuclear inclusion X (NIX) is a gamma proteobacteria that infects the nuclei of gill epithelial cells in Pacific razor clams. NIX has been associated with clam die-offs in coastal Washington. A quantitative PCR (qPCR) assay was developed to detect NIX in Pacific razor clams, and assay specificity was confirmed by chromogenic in situ hybridization (CISH). Both tests were applied to evaluate NIX infections in wild Pacific razor clams collected during spring 2019. Consistent with results from earlier histopathological assessments, qPCR and CISH indicated 100% prevalence in razor clams from two Washington beaches and 0% prevalence from two Alaskan beaches.

Alaska, Washington

Disruption of the Francisella noatunensis orientalis pdpA gene results in virulence attenuation and protection in zebrafish

Several Francisella spp. including F. noatunensis are regarded as important emerging pathogens of wild and farmed fish. However, very few studies have investigated the virulence factors that allow these bacterial species to be pathogenic in fish. The Francisella Pathogenicity Island (FPI) is a well-described, gene-dense region encoding major virulence factors for the genus Francisella. PdpA is a member of the pathogenicity determining protein genes encoded by the FPI that are implicated in the ability of the mammalian pathogen, F. tularensis , to escape and replicate in infected host cells. Using a sacB suicide approach, we generated pdpA knockouts to address the role of PdpA as a virulence factor for F. noatunensis . Because polarity can be an issue in gene-dense regions, we generated two different marker-based mutants in opposing polarity ( Fno Δ pdpA1 and Δ pdpA2 ). Both mutants were attenuated (p<0.0001) in zebrafish challenges and displayed impaired intracellular replication (p<0.05) and cytotoxicity (p<0.05), all of which could be restored to wild-type (WT) levels by complementation for Fno Δ pdpA 1. Importantly, differences were found for bacterial burden and induction of acute phase and pro-inflammatory genes for Fno Δ pdpA 1 and Δ pdpA 2 compared to WT during acute infection. In addition, neither mutant resulted in significant histopathological changes. Finally, immunization with Fno Δ pdpA1 led to protection (p<0.012) against an acute lethal-dose 40 challenge with WT Fno in the zebrafish model of infection. Taken together, this study further demonstrates physiological similarities within the genus Francisella relative to their phylogenetic relationships and the utility of zebrafish for addressing virulence factors for the genus.

Infection and Immunity

Differential susceptibility of Yukon River and Salish Sea stocks of Chinook salmon Oncorhynchus tshawytscha to ichthyophoniasis

Preliminary evidence suggests that Chinook salmon Oncorhynchus tshawytscha from the Yukon River may be more susceptible to Ichthyophonus sp. infections than Chinook from stocks further south. To investigate this hypothesis in a controlled environment, we experimentally challenged juvenile Chinook from the Yukon River and from the Salish Sea with Ichthyophonus sp. and evaluated mortality, infection prevalence and infection load over time. We found that juvenile Chinook salmon from a Yukon River stock were more susceptible to ichthyophoniasis than were those from a Salish Sea stock. After feeding with tissues from infected Pacific herring Clupea pallasii , Chinook salmon from both stocks became infected. The infection was persistent and progressive in Yukon River stock fish, where infections sometimes progressed to mortality, and histological examinations revealed parasite dissemination and proliferation throughout the host tissues. In Salish Sea-origin fish, however, infections were largely transient; host mortalities were rare, and parasite stages were largely cleared from most tissues after 3-4 wk. Susceptibility differences were evidenced by greater cumulative mortality, infection prevalence, parasite density, proportion of fish demonstrating a cellular response, and intensity of the cellular response among fish from the Yukon River stock. These observed differences between Chinook salmon stocks were consistent when parasite exposures occurred in both freshwater and seawater. These results support the hypothesis that a longer-standing host-pathogen relationship, resulting in decreased disease susceptibility, exists among Salish Sea Chinook salmon than among Yukon River conspecifics.

Diseases of Aquatic Organisms

Consequences of Piscine orthoreovirus genotype 1 (PRV‐1) infections in Chinook salmon (Oncorhynchus tshawytscha ), coho salmon (O. kisutch ) and rainbow trout (O. mykiss )

Piscine orthoreovirus genotype 1 (PRV‐1) is the causative agent of heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon ( Salmo salar L.). The virus has also been found in Pacific salmonids in western North America, raising concerns about the risk to native salmon and trout. Here, we report the results of laboratory challenges using juvenile Chinook salmon, coho salmon and rainbow trout injected with tissue homogenates from Atlantic salmon testing positive for PRV‐1 or with control material. Fish were sampled at intervals to assess viral RNA transcript levels, haematocrit, erythrocytic inclusions and histopathology. While PRV‐1 replicated in all species, there was negligible mortality in any group. We observed a few erythrocytic inclusion bodies in fish from the PRV‐1‐infected groups. At a few time points, haematocrits were significantly lower in the PRV‐1‐infected groups relative to controls, but in no case was anaemia noted. The most common histopathological finding was mild, focal myocarditis in both the non‐infected controls and PRV‐1‐infected fish. All cardiac lesions were judged mild, and none were consistent with those of HSMI. Together, these results suggest all three species are susceptible to PRV‐1 infection, but in no case did infection cause notable disease in these experiments.

Journal of Fish Diseases

Mortality of endangered juvenile Lost River Suckers associated with cyanobacteria blooms in mesocosms in Upper Klamath Lake, Oregon

Unsustainably high mortality within the first 2 years of life prevents endangered Lost River Suckers Deltistes luxatus in Upper Klamath Lake, Oregon, from recruiting to spawning populations. Massive blooms of the cyanobacterium Aphanizomenon flos‐aquae and their subsequent death and decay in the lake (bloom‐crashes) are associated with high pH, low percent oxygen saturation, high total ammonia concentrations, and spikes in the cyanotoxin microcystin. Poor water quality within the lake is considered the most likely cause of juvenile sucker mortality, but mechanisms causing the high mortality are not known. We introduced PIT‐tagged age‐1 Lost River suckers into three continuously monitored mesocosms in Upper Klamath Lake to determine the timing of juvenile sucker mortality relative to pH, temperature, and dissolved oxygen. Mortality was inferred from a lack of movement detected on remote PIT tag detection equipment within each mesocosm. Mortality was compared among mesocosms and an indoor tank‐held control group. We fitted time‐varying Cox hazard models to test hypotheses about short‐term and chronic effects of single and co‐occurring water quality parameters on the daily hazard rate. Presumed healthy or moribund fish that were collected pre‐season, mid‐season, or at the end of the study were examined macroscopically and histologically to generate inferences about the causes of mortality. Models did not indicate a plausible association between water quality variables and mortality. Hypoxia preceded periods of higher mortality at two of three sites but did not co‐occur with mortality. Hatchery‐reared Lost River Suckers confined to mesocosms may not represent the behavior of wild fish, and it is unclear whether the same factors affect the mortality of wild age‐0 suckers.

Oregon

Effects of microcystin-LR on juvenile Lost River suckers (Deltistes luxatus) during feeding trials, Upper Klamath Lake, Oregon, 2014−16

Executive Summary Historically, populations of Lost River suckers ( Deltistes luxatus ) of the Upper Klamath Basin were so numerous that they were commercially harvested; however, declining numbers throughout the 20th century led to the listing of the species under the United States Endangered Species Act in 1988. Habitat destruction, poor water quality, competition with (and predation by) nonnative species, especially fathead minnows ( Pimephales promelas ) and yellow perch ( Perca flavescens ), are hypothesized as primary causes of population decline (U.S. Fish and Wildlife Service, 2013). Age data indicate that almost all adult suckers presently in Upper Klamath Lake spawning populations were hatched in the early 1990s. While entrainment of young fish (especially larvae) may contribute, catch-at-length and age data suggest consistently high mortality during the first year of life may be preventing the recruitment of young adults. The specific causes of juvenile sucker mortality are unknown; however, the absence of juvenile suckers in trap net catches coincides with degraded water quality associated with the decay of cyanobacteria blooms and exposure to toxic microcystin produced by Microcystis cyanobacteria. Water-quality data collected in Upper Klamath Lake from 2011 to 2016 suggest that microcystin concentrations in Upper Klamath Lake reached potentially lethal levels based on literature findings from studies on a variety of fish species. We conducted a laboratory feeding trial to determine if microcystin toxicity could potentially be a direct cause of juvenile Lost River sucker mortality. We examined the effects of environmentally relevant doses of microcystin on the survival and health of hatchery-reared juvenile Lost River suckers. Results from this laboratory study suggest that Lost River suckers are very tolerant of the microcystin-LR toxin. Histopathological analysis revealed no evidence of tissue changes associated with microcystin-LR exposure. Although no direct effects of microcystin-LR exposure were detected, suckers could potentially be negatively affected through added energy expenditures and stress associated with excretion of microcystin. Furthermore, microcystin may adversely affect other organisms in Upper Klamath Lake that could alter food availability or habitat of the suckers.

Oregon

Assessing causes of mortality for endangered juvenile Lost River suckers (Deltistes luxatus) in mesocosms in Upper Klamath Lake, south-central Oregon, 2016

Executive Summary The recovery of endangered Lost River suckers ( Deltistes luxatus ) in Upper Klamath Lake, south-central Oregon, has been impeded because juveniles are not recruiting into adult spawning populations. Adult sucker populations spawn each spring but mortality of age-0 suckers during their first summer is excessively high, and recruitment of juveniles into adult populations does not occur in most years. The last significant year class to join spawning aggregations was hatched in 1991. Capture rates for age-0 Lost River suckers decrease so substantially each summer that it is thought that mortality is nearly 100 percent within the first year of life each year. Causes of mortality are not understood but poor water quality, parasites, disease, predation, and non-native species are suspected to contribute to mortality. Upper Klamath Lake is hypereutrophic and summer water-quality conditions have large diurnal and seasonal fluctuations. Photosynthesis of Aphanizomenon flos-aquae , the most abundant cyanobacterium in Upper Klamath Lake, is responsible for large fluctuations in dissolved-oxygen (DO) concentrations and pH. We introduced hatchery-raised, passive integrated transponder-tagged juvenile Lost River suckers into large mesocosms located at Fish Banks, Mid North, and Rattlesnake Point in Upper Klamath Lake, Oregon, to assess sucker mortality relative to water-quality conditions. We identified the date of death for each sucker by assessing movement patterns among vertically stratified antennas. We modeled daily mortality using known fate models relative to water-quality conditions measured by sondes. Histopathology was used to understand causes of eminent mortality for moribund suckers. Fish mortality, growth, health, and movement patterns varied among locations, but it was unclear whether this variation was due to water-quality or other factors. Seasonal mortality was 58.8 percent at Fish Banks, 27.4 percent at Mid North, and 11.5 percent at Rattlesnake Point. Growth over the 109-day study period was lowest at Fish Banks (34.5 ±10.0 millimeters [mm] standard length (SL); 18.6 ±7.7 grams [g]), intermediate at Mid North (57.5 ±13.6 mm SL; 40.1 ±15.4 g), and greatest at Rattlesnake Point (78.4 ±13.0 mm SL; 72.5 ±18.7 g). Our ability to assess causes of juvenile sucker mortality in mesocosms using our modelling approach was limited by low daily mortality. Zero to 3 mortalities occurred per day, except on July 30 at Fish Banks when 7 mortalities occurred. Relative to any other measured and tested water-quality condition, mortality was more likely to occur on days with large fluctuations in oxygen percent saturation. When we assessed the fit of the most parsimonious model, performance was poor, which suggested that other factors were contributing to mortality. Our ability to assess the relationship between seasonal patterns in water quality and fish mortality were limited by the absence of substantial differences in water quality among sites, inconsistency in the depth at which measurements were collected, and no clear pattern in conditions leading up to and during mortality events. Except for DO at Rattlesnake Point and diel temperature variations at Fish Banks, seasonally summarized water-quality factors were similar among sites. The locations of water-quality monitors within the water column likely explain the differences in DO at Rattlesnake Point and temperature variation at Fish Banks. Furthermore, DO concentrations and other water-quality factors occurring during and prior to mortality events were inconsistent. Microscopic assessments indicated severe gill hyperplasia, fusion of the secondary lamellae, and severe Ichthyobodo sp. infestations on the gills of most moribund suckers. Liver glycogen was usually depleted in suckers with severe Ichthyobodo sp. infestations. Ichthyobodo sp. infestations probably were the immediate cause of death and probably originated from the Klamath Tribes Fish Research Facility, although this parasite also is present in Upper Klamath Lake and severe water-quality conditions may have contributed to morbidity. As suckers in the mesocosms died, they were replaced with suckers from the Fish Research Facility that likely were heavily parasitized with Ichthyobodo sp. Therefore, it is possible that the gradient in mortality rate among sites was owing to site-varying differences in inadvertent increases in introduced parasite loads.

Oregon

Health and condition of endangered young-of-the-year Lost River and Shortnose suckers relative to water quality in Upper Klamath Lake, Oregon, 2014–2015

Most mortality of endangered Lost River ( Deltistes luxatus ) and shortnose ( Chasmistes brevirostris ) suckers in Upper Klamath Lake, Oregon, occurs within the first year of life. Juvenile suckers in Clear Lake Reservoir, California, survive longer and may even recruit to the spawning populations. In a previous (2013–2014) study, the health and condition of juvenile suckers and the dynamics of water quality between Upper Klamath Lake and Clear Lake Reservoir were compared. That study found that apparent signs of stress or exposure to irritants, such as peribiliary cuffing in liver tissue and mild inflammation and necrosis in gill tissues, were present in suckers from both lakes and were unlikely to be clues to the cause of differential mortality between lakes. Seasonal trends in energy storage as glycogen and triglycerides were also similar between lakes, indicating prey limitation was not a likely factor in differential mortality. To better understand the relationship between juvenile sucker health and water quality, we examined suckers collected in 2014–2015 from Upper Klamath Lake, where water quality can be dynamic and, at times, extreme. While there were notable differences in water quality and fish health between years, we were not able to identify any specific water-quality-related causes for differential fish condition. Water quality was generally better in 2014 than in 2015. When considered together afflictions and abnormalities generally indicated healthier suckers in 2014 than 2015. Low dissolved-oxygen events (< 4 milligrams per liter) were less frequent and occurred earlier; high pH events (≥ 9.5) were less frequent and shorter in duration; large diel fluctuations in pH (≥ 1.4) were less frequent; water temperatures were warmer, particularly in July and September; and concentrations of microcystin in both large and small fractions of samples were lower in 2014 than in 2015. Total and therefore also un-ionized ammonia were low in 2014–2015 relative to concentrations known to affect suckers. Petechial hemorrhages of the skin, attached Lernaea spp. and eosinophilic hyaline droplets in the kidney tubules were less prevalent in 2014 than in 2015; however, hyperplastic and hypertrophic gill tissue and trichodinids on the gills were observed more frequently in 2014. There were more suckers with normal liver color and texture in 2014 than in 2015. The prevalence of suckers with liver inflammation was greater in 2014 and only observed in suckers collected after August 5, whereas liver inflammation occurred intermittently in 2015. Liver glycogen among suckers decreased in late-August 2014 and increased from early August to mid-September 2015. Lost River suckers had greater whole-body triglyceride content but a larger proportion with an absence of visceral fat observed in 2014 than in 2015. In contrast, shortnose suckers were similar between years in regard to both whole-body triglyceride and visceral fat. Black-spot-forming parasites (trematode metacercariae) were observed in a higher prevalence on shortnose suckers but not Lost River suckers in 2014 than in 2015. Opercular deformities were less prevalent in both species in 2014 than in 2015. Neither gross nor histological examination revealed a high prevalence of abnormalities in suckers that clearly indicate a primary mechanism for juvenile mortality in Upper Klamath Lake. Histological abnormalities were almost always focal and minimal or mild except where associated with parasites. Mild to severe focal abnormalities associated with Lernaea sp. attachment sites and encysted digenean (trematode) metacercariae are unlikely to be associated with mortality. Severe and diffuse inflammation and hyperplasia of the gills associated with Ichthyobodo sp. on one Lost River sucker, may indicate a potential cause of mortality. High mortality may have primarily occurred outside our study period (for example, in spring or over winter), or was caused by a factor that could not be detected with our methods (for example, predation). Alternatively, abnormalities in a small percentage of passively captured suckers in Upper Klamath Lake may indicate health-related issues that were more prevalent in populations than in our samples. Temporary decreases in liver glycogen stores may also indicate periods of stress, which may eventually lead to mortality of young suckers.

Oregon

Pufferfish mortality associated with novel polar marine toxins in Hawaii

Fish die-offs are important signals in tropical marine ecosystems. In 2010, a mass mortality of pufferfish in Hawaii (USA) was dominated by Arothron hispidus showing aberrant neurological behaviors. Using pathology, toxinology, and field surveys, we implicated a series of novel, polar, marine toxins as a likely cause of this mass mortality. Our findings are striking in that (1) a marine toxin was associated with a kill of a fish species that is itself toxic; (2) we provide a plausible mechanism to explain clinical signs of affected fish; and (3) this epizootic likely depleted puffer populations. Whilst our data are compelling, we did not synthesize the toxin de novo, and we were unable to categorically prove that the polar toxins caused mortality or that they were metabolites of an undefined parent compound. However, our approach does provide a template for marine fish kill investigations associated with marine toxins and inherent limitations of existing methods. Our study also highlights the need for more rapid and cost-effective tools to identify new marine toxins, particularly small, highly polar molecules.

Hawaii

Survival, movement, and health of hatchery-raised juvenile Lost River suckers within a mesocosm in Upper Klamath Lake, Oregon

The recovery of endangered Lost River suckers ( Deltistes luxatus ) in Upper Klamath Lake is limited by poor juvenile survival and failure to recruit into the adult population. Poor water quality, degradation of rearing habitat, and toxic levels of microcystin are hypothesized to contribute to low juvenile survival. Studies of wild juvenile suckers are limited in that capture rates are low and compromised individuals are rarely captured in passive nets. The goal of this study was to assess the use of a mesocosm for learning about juvenile survival, movement, and health. Hatchery-raised juvenile Lost River suckers were PIT (passive integrated transponder) tagged and monitored by three vertically stratified antennas. Fish locations within the mesocosm were recorded at least every 30 minutes and were assessed in relation to vertically stratified water-quality conditions. Vertical movement patterns were analyzed to identify the timing of mortality for each fish. Most mortality occurred from July 28 to August 16, 2014. Juvenile suckers spent daylight hours near the benthos and moved throughout the entire water column during dark hours. Diel movements were not in response to dissolved-oxygen concentrations, temperature, or pH. Furthermore, low dissolved-oxygen concentrations, high temperatures, high pH, high un-ionized ammonia, or high microcystin levels did not directly cause mortality, although indirect effects may have occurred. However, water-quality conditions known to be lethal to juvenile Lost River suckers did not occur during the study period. Histological assessment revealed severe gill hyperplasia and Ichthyobodo sp. infestations in most moribund fish. For these fish, Ichthyobodo sp. was likely the cause of mortality, although it is unclear if this parasite originated in the rearing facility because fish were not screened for this parasite prior to introduction. This study has demonstrated that we can effectively use a mesocosm equipped with antennas to learn about the timing of mortality, movement, and health of PIT-tagged hatchery-raised juvenile Lost River suckers.

Oregon

Experimental infection of six North American fish species with the North Carolina strain of spring Viremia of Carp Virus

Spring viremia of carp virus (SVCV) is a rhabdoviral pathogen associated with disease outbreaks in cultured and wild fish worldwide. Common carp ( Cyprinus carpio carp ), and koi ( C. carpio koi ) suffer the highest mortalities from SVCV infections, while other cyprinid fish species have varying susceptibility. Although salmonid fish typically are considered refractory to infection by SVCV, there have been a few reports suggesting infection has occurred in rainbow trout ( Oncorhynchus mykiss ). There have been no reports of Percid fish being infected with SVCV. Since the first North American outbreak of SVCV at a North Carolina koi farm in 2002 there have been eight subsequent detections or outbreaks of SVCV among fish species from the families of Cyprinidae and Centrarchidae within the US and Canada. Thus, this exotic virus is considered a potential threat to native and cultured fish populations in North America. We performed multiple experimental challenges with fish species from three families ( Salmonidae , Cyprinidae , and Percidae ) to identify the potential risk associated with SVCV exposure of resident fish populations in North America. Three salmonid species, rainbow and steelhead trout ( Oncorhynchus mykiss ), Chinook salmon ( O. tshawytscha ), and sockeye salmon ( O. nerka ), were challenged by immersion or injection with the North Carolina SVCV isolate. Two cyprinid species, koi and fathead minnow ( Pimephales promelas ) and one percid species, yellow perch ( Perca flavescens ) were also challenged. Koi were highly susceptible to SVCV up to 11 months of age and fathead minnows had chronic disease expression with moderate mortality (29%). SVCV also induced moderate mortalities (33%) in yellow perch fry. Virus challenged salmonid fish had cumulative percent mortalities ranging from 0 to 100%, with sockeye salmon fry being the most vulnerable. A sub-sample of mortalities and survivors were screened for virus by plaque assay and reverse transcription polymerase chain reaction. In general, all mortalities tested positive for SVCV with high viral titers while survivors had variable persistence of SVCV with overall lower virus titers. Our SVCV challenges of multiple North American fish species suggested that host age is a key factor in determining disease outcome. Other factors, such as fish broodstock, virus strain, water temperature, and rearing conditions in association with the intrinsic level of species susceptibility may also impact infection dynamics. This is the first report of SVCV infecting a species (yellow perch) in the family Percidae and that sockeye salmon fry can suffer similarly high mortalities as the primary SVCV host species.

Aquaculture

Infecting Pacific Herring with Ichthyophonus sp. in the laboratory

The protistan parasite Ichthyophonus sp. occurs in coastal populations of Pacific Herring Clupea pallasii throughout the northeast Pacific region, but the route(s) by which these planktivorous fish become infected is unknown. Several methods for establishing Ichthyophonus infections in laboratory challenges were examined. Infections were most effectively established after intraperitoneal (IP) injections with suspended parasite isolates from culture or after repeated feedings with infected fish tissues. Among groups that were offered the infected tissues, infection prevalence was greater after multiple feedings (65%) than after a single feeding (5%). Additionally, among groups that were exposed to parasite suspensions prepared from culture isolates, infection prevalence was greater after exposure by IP injection (74%) than after exposure via gastric intubation (12%); the flushing of parasite suspensions over the gills did not lead to infections in any of the experimental fish. Although the consumption of infected fish tissues is unlikely to be the primary route of Ichthyophonus sp. transmission in wild populations of Pacific Herring, this route may contribute to abnormally high infection prevalence in areas where juveniles have access to infected offal.

Journal of Aquatic Animal Health