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Rachel L. Powers

Publications and source records attributed to Rachel L. Powers.

11 recordsLinked to original sources

Local and systemic replicative fitness for viruses in specialist, generalist, and non-specialist interactions with salmonid hosts

Host tissues represent diverse resources or barriers for pathogen replicative fitness. We tested whether viruses in specialist, generalist, and non-specialist interactions replicate differently in local entry tissue (fin), and systemic target tissue (kidney) using infectious hematopoietic necrosis virus (IHNV) and three salmonid fish hosts. Virus tissue replication was host specific, but one feature was shared by specialists and the generalist which was uncommon in the non-specialist interactions: high host entry and replication capacity in the local tissue after contact. Moreover, specialists showed increased replication in systemic target tissues early after host contact. By comparing ancestral and derived IHNV viruses, we also characterized replication tradeoffs associated with specialist and generalist evolution. Compared with the ancestral virus, a derived specialist gained early local replicative fitness in the new host but lost replicative fitness in the ancestral host. By contrast, a derived generalist showed small replication losses relative to the ancestral virus in the ancestral host but increased early replication in the local tissue of novel hosts. This study shows that the mechanisms of specialism and generalism are host specific and that local and systemic replication can contribute differently to overall within host replicative fitness for specialist and generalist viruses.

Journal of General Virology

Rapid diagnostic test to detect and discriminate infectious hematopoietic necrosis virus (IHNV) genogroups U and M to aid management of Pacific Northwest salmonid populations

Infectious hematopoietic necrosis virus (IHNV) is an acute pathogen of salmonids in North America, Europe, and Asia that is phylogenetically classified into five major virus genogroups (U, M, L, E, and J). The geographic range of the U and M genogroup isolates overlap in the North American Columbia River Basin and Washington Coast region, where these genogroups pose different risks depending on the species of Pacific salmon ( Oncorhynchus spp.). For certain management decisions, there is a need to both test for IHNV presence and rapidly determine the genogroup. Herein, we report the development and validation of a U/M multiplex reverse transcription, real-time PCR (RT-rPCR) assay targeting the IHNV nucleocapsid (N) protein gene. The new U/M RT-rPCR is a rapid, sensitive, and repeatable assay capable of specifically discriminating between North American U and M genogroup IHNV isolates. However, one M genogroup isolate obtained from commercially cultured Idaho rainbow trout ( O. mykiss ) showed reduced sensitivity with the RT-rPCR test, suggesting caution may be warranted before applying RT-rPCR as the sole surveillance test in areas associated with the Idaho trout industry. The new U/M assay had high diagnostic sensitivity (DSe > 94%) and specificity (DSp > 97%) in free-ranging adult Pacific salmon, when assessed relative to cell culture, the widely accepted reference standard, as well as the previously validated universal N RT-rPCR test. The high diagnostic performance of the new U/M assay indicates the test is suitable for surveillance, diagnosis, and confirmation of IHNV in Pacific salmon from the Pacific Northwest regions where the U and M genogroups overlap.

Animals

A phylogeny based on cytochrome-c oxidase gene sequences identifies sympatric Ichthyophonus genotypes in the NE Pacific Ocean

ABSTRACT: In recent decades, evidence has accumulated to suggest that the widespread and highly variable parasite Ichthyophonus hoferi is actually a species complex. Highly plastic morphology and a general lack of defining structures has contributed to the likely underestimate of biodiversity within this group. Molecular methods are a logical next step in the description of these parasites, but markers used to date have been too conserved to resolve species boundaries. Here we use mitochondrial encoded cytochrome-c oxidase (MTCO1) gene sequences and phylogenic analysis to compare Ichthyophonus spp. isolates from several marine and anadromous fish hosts. The resulting phylogeny displays lineage separation among isolates and possible host/niche segregation not previously described. The parasite type that infects Pacific herring Clupea pallasii , Atlantic herring C. harengus , Atlantic salmon Salmo salar , and Pacific staghorn sculpin Oligocottus maculosus (Clade A) is different from that which infects Chinook salmon Oncorhynchus tshawytscha, walleye pollock Gadus chalcogrammus , Greenland halibut Reinhardtius hippoglossoides , and Pacific halibut Hippoglossus stenolepsis (Clade B). MTCO1 sequences confirmed the presence of a more divergent Ichthyophonus sp. isolated from American shad Alosa sapidissima in rivers of eastern North America (Clade C), while American shad introduced to the Pacific Ocean are infected with the same parasite that infects Pacific herring (Clade A). Currently there are no consensus criteria for delimiting species within Ichthyophonidae , but MTCO1 sequences hold promise as a potential species identifying marker and useful epizootiological tool.

Diseases of Aquatic Organisms

Temperature variation and host immunity regulate viral persistence in a salmonid host

Environmental variation has important effects on host–pathogen interactions, affecting large-scale ecological processes such as the severity and frequency of epidemics. However, less is known about how the environment interacts with host immunity to modulate virus fitness within hosts. Here, we studied the interaction between host immune responses and water temperature on the long-term persistence of a model vertebrate virus, infectious hematopoietic necrosis virus (IHNV) in steelhead trout ( Oncorhynchus mykiss ). We first used cell culture methods to factor out strong host immune responses, allowing us to test the effect of temperature on viral replication. We found that 15 ∘ "> ∘ C water temperature accelerated IHNV replication compared to the colder 10 and 8 ∘ "> ∘ C temperatures. We then conducted in vivo experiments to quantify the effect of 6, 10, and 15 ∘ "> ∘ C water temperatures on IHNV persistence over 8 months. Fish held at 15 and 10 ∘ "> ∘ C were found to have higher prevalence of neutralizing antibodies compared to fish held at 6 ∘ "> ∘ C. We found that IHNV persisted for a shorter time at warmer temperatures and resulted in an overall lower fish mortality compared to colder temperatures. These results support the hypothesis that temperature and host immune responses interact to modulate virus persistence within hosts. When immune responses were minimized (i.e., in vitro) virus replication was higher at warmer temperatures. However, with a full potential for host immune responses (i.e., in vivo experiments) longer virus persistence and higher long-term virulence was favored in colder temperatures. We also found that the viral RNA that persisted at later time points (179 and 270 days post-exposure) was mostly localized in the kidney and spleen tissues. These tissues are composed of hematopoietic cells that are favored targets of the virus. By partitioning the effect of temperature on host and pathogen responses, our results help to better understand environmental drivers of host–pathogen interactions within hosts, providing insights into potential host–pathogen responses to climate change.

Pathogens

Effect of temperature on survival of Lost River Suckers with a natural infection of Ichthyobodo spp.

To compensate for low natural survival of endangered Lost River Suckers Deltistes luxatus , the U.S. Fish and Wildlife Service and the Klamath Tribes have initiated captive rearing programs. We conducted laboratory experiments intended to determine the temperature for optimum growth of juvenile Lost River Suckers; however, due to an unanticipated infection with Ichthyobodo spp., we instead estimated survival in conjunction with temperature and parasite loads. Ichthyobodo spp. are common parasites that infest fish skin and gills; they are known to be present in Upper Klamath Lake and have been found in fish at both the federal and tribal fish-rearing facilities. All fish held at mean temperatures of 16°C and 19°C survived the entire 65-d experiment, whereas the median time to death was 27 d at 22°C, 8 d at 24°C, and 7 d at 26°C. Gill samples from all Lost River Suckers tested positive for Ichthyobodo spp. DNA, with a trend toward higher copy numbers in suckers that were held at a mean temperature of 22°C or warmer compared to those held at 19°C or cooler. Fish kept at all temperatures grew slowly. These results suggest that survival of Lost River Suckers with a natural infection of Ichthyobodo spp. was significantly impacted by temperature.

North American Journal of Aquaculture

Long-term shedding from fully convalesced individuals indicates that Pacific herring are a reservoir for viral hemorrhagic septicemia virus

Processes that allow viral hemorrhagic septicemia (VHS) virus to persist in the marine environment remain enigmatic, owing largely to the presence of covert and cryptic infections in marine fishes during typical sub-epizootic periods. As such, marine host reservoirs for VHS virus have not been fully demonstrated, nor have the mechanism(s) by which infected hosts contribute to virus perpetuation and transmission. Here, we demonstrate that after surviving VHS, convalesced Pacific herring continue to shed virus at a low rate for extended periods. Further, exposure of previously naïve conspecific sentinels to this shed virus can result in infections for at least 6 mo after cessation of overt disease. This transmission mechanism was not necessarily dependent on the magnitude of the disease outbreak, as prolonged transmission occurred from 2 groups of donor herring that experienced cumulative mortalities of 4 and 29%. The results further suggest that the virus persists in association with the gills of fully recovered individuals, and long-term viral shedding or shedding relapses are related to cooler or decreasing water temperatures. These results provide support for a new VHS virus perpetuation paradigm in the marine environment, whereby the virus can be maintained in convalesced survivors and trafficked from these carriers to sympatric susceptible individuals.

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

Broad-spectrum antiviral JL122 blocks infection and inhibits transmission of aquatic rhabdoviruses

The aquaculture industry is growing rapidly to meet the needs for global protein consumption. Viral diseases in aquaculture are quite challenging due to lack of treatment options as well as limited injection-delivery vaccines, which are costly. Thus, water-immersion antiviral treatments are highly desirable. This study focused on broad-spectrum, light-activated antivirals that target the viral membrane (envelope) of viruses to prevent viral-cell membrane fusion, ultimately blocking viral entry into cells. Of the tested small-molecules, JL122, a new broad-spectrum antiviral previously unexplored against aquatic viruses, blocked infection of three aquatic rhabdoviruses (IHNV, VHSV and SVCV) in cell culture and in two live fish challenge models. Importantly, JL122 inhibited transmission of IHNV from infected to uninfected rainbow trout. Further, the effective antiviral concentrations were not toxic to cells or susceptible fish. These results show promise for JL122 to become an immersion treatment option for outbreaks of aquatic enveloped viral infections.

Virology

Detection of Nanophyetus salmincola in water, snails, and fish tissues by quantitative polymerase chain reaction

We report the development and validation of two quantitative PCR (qPCR) assays to detect Nanophyetus salmincola DNA in water samples and in fish and snail tissues. Analytical and diagnostic validation demonstrated good sensitivity, specificity, and repeatability of both qPCR assays. The N. salmincola DNA copy number in kidney tissue was significantly correlated with metacercaria counts based on microscopy. Extraction methods were optimized for the sensitive qPCR detection of N. salmincola DNA in settled water samples. Artificially spiked samples suggested that the 1-cercaria/L threshold corresponded to an estimated log 10 copies per liter ≥ 6.0. Significant correlation of DNA copy number per liter and microscopic counts indicated that the estimated qPCR copy number was a good predictor of the number of waterborne cercariae. However, the detection of real-world samples below the estimated 1-cercaria/L threshold suggests that the assays may also detect other N. salmincola life stages, nonintact cercariae, or free DNA that settles with the debris. In summary, the qPCR assays reported here are suitable for identifying and quantifying all life stages of N. salmincola that occur in fish tissues, snail tissues, and water.

Journal of Aquatic Animal Health