Geology topics
Diane G. Elliott
Publications and source records attributed to Diane G. Elliott.
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.
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.
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.
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.
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.
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.
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.
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.
In memoriam - William Toshio (Tosh) Yasutake, 1922-2016
William Toshio (Tosh) Yasutake, 1922-2016 passed away peacefully at home on December 12, 2016, at the age of 94. He is survived by Fumi, his wife of 66 years, as well as four children and six grandchildren. With his death, the fish health community has lost an outstanding scientist as well as a kind, unassuming, and wonderful human being. Tosh was born on June 10, 1922, in Seattle, Washington, to Jack and Hide Yasutake. He was in his first year of studies at the University of Washington when Pearl Harbor was attacked by Imperial Japan on December 7, 1941. Following the attack, Tosh and his family (father, mother, sister, and two brothers) were among the 110,000–120,000 people of Japanese ancestry who were forced from their homes on the Pacific coast and incarcerated in internment camps in the interior. In June 1942, Tosh enlisted in the U.S. Army, serving as an unarmed combat medic in the famed 442nd Regimental Combat Team, the most decorated unit for its size and length of service in the history of American warfare. Wounded in October 1944 during the Vosges Mountains campaign near Bruyères, France, Tosh was evacuated and missed the ensuing battle to rescue the “Lost Battalion,” at which his replacement was killed. Tosh returned to action in Italy in February 1945 and served until the end of the war in Europe, earning both a Purple Heart and a Bronze Star for bravery. In October 2010, the Congressional Gold Medal was awarded to the 442nd Regimental Combat Team, and in 2012 the surviving members were made chevaliers of the French Légion d’Honneur for actions contributing to the liberation of France in World War II. After the war, Tosh returned to the University of Washington on the GI Bill and received a B.S. degree in zoology in 1951. In 1953 he began his research career at the U.S. Fish and Wildlife Service’s Western Fish Nutrition Laboratory at Cook, Washington, where he conducted pioneering research on nutritional fish diseases with John Halver. Tosh was one of the first to recognize hepatomas in hatchery-reared Rainbow Trout and helped to trace the disease to an aflatoxin produced by the mold Aspergillus flavis , which grew during the storage of ingredients for fish diets. In 1960, he transferred to the Western Fisheries Research Center (WFRC) in Seattle (then called the Western Fish Disease Laboratory) to start a fish pathology diagnostic laboratory. There he described the histopathology of diseases of economically important fishes, identified etiologic agents, and worked with hatchery biologists to improve the health, quality, and survival of salmonids released from federal and state hatcheries. Tosh was instrumental in recognizing that the viruses of Oregon sockeye disease and Chinook Salmon virus disease were one entity and in giving the disease its present name: infectious hematopoietic necrosis. In recognition of his pioneering research, Tosh was awarded a doctorate in fish pathology by the University of Tokyo in 1980, the first American to have been so honored. In 1983, he published his classic textbook The Microscopic Anatomy of Salmonids: An Atlas , which quickly became a standard reference work in fish pathology and is still in wide use today. For his outstanding career achievements, in 1987 Tosh received the S. F. Snieszko Distinguished Service Award, the highest honor bestowed by the American Fisheries Society’s Fish Health Section (AFS–FHS). Tosh retired in 1988 but continued his research at the WFRC as a senior scientist emeritus, providing technical assistance to federal and state agencies and to the aquaculture industry worldwide. His culminating project was to digitize his lifetime collection of photomicrographs and prepare an atlas, “Histopathology of Selected Parasitic Salmonid Diseases: A Color Atlas,” that is now posted on the Web sites of the WFRC and the AFS–FHS. Although his presence will be sorely missed, his research contributions have become part of the foundation of today’s knowledge of fisheries biology and have assured him a place in history.
Detecting Renibacterium salmoninarum in wild brown trout by use of multiple organ samples and diagnostic methods
Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease (BKD), is endemic in many wild trout species in northerly regions. The aim of the present study was to determine the optimal R. salmoninarum sampling/testing strategy for wild brown trout (Salmo trutta L.) populations in Iceland. Fish were netted in a lake and multiple organs—kidney, spleen, gills, oesophagus and mid-gut—were sampled and subjected to five detection tests i.e. culture, polyclonal enzyme-linked immunosorbent assay (pELISA) and three different PCR tests. The results showed that each fish had encountered R. salmoninarum but there were marked differences between results obtained depending on organ and test. The bacterium was not cultured from any kidney sample while all kidney samples were positive by pELISA. At least one organ from 92.9% of the fish tested positive by PCR. The results demonstrated that the choice of tissue and diagnostic method can dramatically influence the outcome of R. salmoninarum surveys.
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.
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.
Health and condition of endangered juvenile Lost River and shortnose suckers relative to water quality and fish assemblages in Upper Klamath Lake, Oregon, and Clear Lake Reservoir, California
Executive Summary Most mortality of endangered Lost River ( Deltistes luxatus ) and shortnose ( Chasmistes brevirostris ) suckers in Upper Klamath Lake, Oregon, appears to occur within the first year of life. However, juvenile suckers in Clear Lake Reservoir, California, appear to survive longer and may even recruit to the spawning populations. Our goal in this study was to develop productive lines of inquiry into the causes of mortality of juvenile suckers, especially in Upper Klamath Lake, through comparison of sucker health and environmental conditions in both lakes. The health of juvenile suckers was associated with physical, biological, and chemical characteristics in each lake from July to September 2013 and 2014. Water-quality dynamics differed substantially between lakes. Diel fluctuations were greater for dissolved-oxygen concentrations and pH in Upper Klamath Lake than in Clear Lake Reservoir, but diel temperature fluctuations were greater in Clear Lake Reservoir. Minimum dissolved-oxygen concentrations were as low as 1.0 milligram per liter (mg/L) in Upper Klamath Lake, but were no lower than 5.7 mg/L in Clear Lake Reservoir. Unionized ammonia (NH 3 ) concentrations were near or less than the minimum reporting limit of 0.002 mg/L NH 3 in Clear Lake Reservoir and generally were less than the concentration known to cause cellular changes to the gills or mortality of suckers in Upper Klamath Lake. Concentrations of microcystins were less than the detection limit (≤0.10 microgram per liter [μg/L]) in both dissolved (<63 micrometers [μm]) and particulate (≥63 μm) fractions of water samples collected from Clear Lake Reservoir in both years. In Upper Klamath Lake, concentrations of microcystins were relatively low in the 2013 particulate fractions and dissolved fractions of samples in both years, but as high as 44.30 μg/L, more than 44 times the World Health Organization recommended levels for drinking water (1 μg/L), in the 2014 particulate fraction. However, there was little histological evidence and no chemical evidence that these concentrations in water were harmful to suckers that we captured. The species and age compositions of juvenile sucker populations differed between lakes. A total of 98 percent of all juvenile suckers captured in Upper Klamath Lake were age-0. In contrast, as many as six age classes of suckers were represented in Clear Lake Reservoir, indicating much better juvenile survival than in Upper Klamath Lake. Based on genetic species identification, 98 percent of juvenile suckers collected from Clear Lake Reservoir were shortnose or Klamath largescale suckers ( Catostomus snyderi ). In contrast, there was a high degree of apparent genetic hybridization in juvenile suckers collected from Upper Klamath Lake, and both Lost River and non-Lost River juvenile suckers were nearly equally represented in samples. Neither gross nor histological examination revealed a high prevalence of abnormalities in suckers that might indicate a mechanism for juvenile mortality in Upper Klamath Lake. Therefore, high mortality primarily may have occurred outside our study period (for example, in spring or over winter), or was owing to 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. Some apparent symptoms of stress or exposure to irritants, such as peribiliary cuffing in hepatocytes and mild inflammation and necrosis in gill tissues, were present in suckers from both lakes and do not appear to be clues to the cause of differential mortality between lakes. Seasonal trends in energy storage as glycogen and triglycerides were similar between lakes, indicating prey availability was not a factor in differential mortality. Differences in sucker health and condition between lakes were considered the most promising clues to the causes of differential juvenile sucker morality between lakes. A low prevalence of petechial hemorrhaging of the skin (16 percent) and deformed opercula (8 percent) in Upper Klamath Lake suckers may indicate exposure to a toxin other than microcystin. Suckers grew slower in their first year of life, but had similar or greater triglyceride and glycogen levels in Upper Klamath Lake compared to Clear Lake Reservoir. These findings do not suggest a lack of prey quantity but may indicate lower prey quality in Upper Klamath Lake.
Effects of temperature on Renibacterium salmoninarum infection and transmission potential in Chinook salmon, Oncorhynchus tshawytscha (Walbaum)
Renibacterium salmoninarum is a significant pathogen of salmonids and the causative agent of bacterial kidney disease (BKD). Water temperature affects the replication rate of pathogens and the function of the fish immune system to influence the progression of disease. In addition, rapid shifts in temperature may serve as stressors that reduce host resistance. This study evaluated the effect of shifts in water temperature on established R. salmoninarum infections. We challenged Chinook salmon with R. salmoninarum at 12°C for 2 weeks and then divided the fish into three temperature groups (8, 12 and 15°C). Fish in the 8°C group had significantly higher R. salmoninarum -specific mortality, kidney R. salmoninarum loads and bacterial shedding rates relative to the fish held at 12 or 15°C. There was a trend towards suppressed bacterial load and shedding in the 15°C group, but the results were not significant. Bacterial load was a significant predictor of shedding for the 8 and 12°C groups but not for the 15°C group. Overall, our results showed little effect of temperature stress on the progress of infection, but do support the conclusion that cooler water temperatures contribute to infection progression and increased transmission potential in Chinook salmon infected with R. salmoninarum .
Persistence of external signs in Pacific herring Clupea pallasii Valenciennes with ichthyophoniasis
The progression of external signs of Ichthyophonus infection in Pacific herring Clupea pallasii Valenciennes was highly variable and asynchronous after intraperitoneal injection with pure parasite preparations; however, external signs generally persisted through the end of the study (429 days post-exposure). Observed signs included papules, erosions and ulcers. The prevalence of external signs plateaued 35 days post-exposure and persisted in 73–79% of exposed individuals through the end of the first experiment (147 days post-exposure). Among a second group of infected herring, external signs completely resolved in only 10% of the fish after 429 days. The onset of mortality preceded the appearance of external signs. Histological examination of infected skin and skeletal muscle tissues indicated an apparent affinity of the parasite for host red muscle. Host responses consisted primarily of granulomatous inflammation, fibrosis and necrosis in the skeletal muscle and other tissues. The persistence and asynchrony of external signs and host response indicated that they were neither a precursor to host mortality nor did they provide reliable metrics for hindcasting on the date of exposure. However, the long-term persistence of clinical signs in Pacific herring may be useful in ascertaining the population-level impacts of ichthyophoniasis in regularly observed populations.
Detection of Ichthyophonus by chromogenic in situ hybridization
Ichthyophonus hoferi (Plehn & Mulsow 1911 ) is a protistan parasite in the class Mesomycetozoea that infects a large range of marine and freshwater fish (Mendoza, Taylor & Ajello 2002 ; McVicar 2011 ). The broad host and geographic range, which includes both fresh and marine waters of the Northern and Southern Hemispheres, combined with a lack of distinguishing morphological characteristics, have prompted speculation that Ichthyophonus -like organisms in multiple species of fish, as well as reptiles, amphibians, birds and invertebrates, may have been incorrectly classified under a single type species I. hoferi (McVicar 2011 ). At present, only two species, I. hoferi and I. irregularis , are currently recognized within the genus (Rand et al . 2000 ; Mendoza et al . 2002 ). Investigations of ribosomal DNA sequence variation have begun to clarify relationships among Ichthyophonus types (Criscione et al . 2002 ; Rasmussen et al . 2010 ). Here, we will use the term Ichthyophonus to broadly represent all members of the genus regardless of species/subspecies.