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Richard Kocan

Publications and source records attributed to Richard Kocan.

6 recordsLinked to original sources

Evidence for an amoeba-like infectious stage of ichthyophonus sp. and description of a circulating blood stage: a probable mechanism for dispersal within the fish host

Small amoeboid cells, believed to be the infectious stage of Ichthyophonus sp., were observed in the bolus (stomach contents) and tunica propria (stomach wall) of Pacific staghorn sculpins and rainbow trout shortly after they ingested Ichthyophonus sp.–infected tissues. By 24–48 hr post-exposure (PE) the parasite morphed from the classically reported multinucleate thick walled schizonts to 2 distinct cell types, i.e., a larger multinucleate amoeboid cell surrounded by a narrow translucent zone and a smaller spherical cell surrounded by a “halo” and resembling a small schizont. Both cell types also appeared in the tunica propria, indicating that they had recently penetrated the columnar epithelium of the stomach. No Ichthyophonus sp. pseudo-hyphae (“germination tubes”) were observed in the bolus or penetrating the stomach wall. Simultaneously, Ichthyophonus sp. was isolated in vitro from aortic blood, which was consistently positive from 6 to 144 hr PE, then only intermittently for the next 4 wk. Small PAS-positive cells observed in blood cultures grew into colonies consisting of non-septate tubules (pseudo-hyphae) terminating in multinucleated knob-like apices similar to those seen in organ explant cultures. Organ explants were culture positive every day; however, typical Ichthyophonus sp. schizonts were not observed histologically until 20–25 days PE. From 20 to 60 days PE, schizont diameter increased from ≤25 μm to ≥82 μm. Based on the data presented herein, we are confident that we have resolved the life cycle of Ichthyophonus sp. within the piscivorous host.

Journal of Parasitology

Synchronous cycling of Ichthyophoniasis with Chinook salmon density revealed during the annual Yukon River spawning migration

Populations of Chinook salmon Oncorhynchus tshawytscha in the Yukon River declined by more than 57% between 2003 and 2010, probably the result of a combination of anthropogenic and environmental factors. One possible contributor to this decline is Ichthyophonus , a mesomycetozoan parasite that has previously been implicated in significant losses of fish, including Chinook salmon. A multiyear epidemiological study of ichthyophoniasis in the Yukon River revealed that disease prevalence and Chinook salmon population abundance increased and decreased simultaneously (i.e., were concordant) from 1999 to 2010. The two values rose and fell synchronously 91% of the time for female Chinook salmon and 82% of the time for males; however, there was no significant correlation between Ichthyophonus prevalence and population abundance. This synchronicity might be explained by a single factor, such as a prey item that is critical to Chinook salmon survival as well as a source of Ichthyophonus infection. The host–parasite relationship between Ichthyophonus and migrating Chinook salmon from 2004 to 2010 was similar to that reported for the previous 5 years. During 2004–2010, overall disease prevalence was significantly higher among females (21%) than among males (8%), increased linearly with fish length for both males and females, and increased in both sexes as the fish progressed upriver. These regularly occurring features of host–parasite dynamics confirm a stable base of transmission for Ichthyophonus . However, from 2003 to 2010, disease prevalence decreased from 30% to just 8% in males and from 45% to 9% in females, paralleling a similar decline in Chinook salmon abundance during the same period. These findings may help clarify questions regarding the complex host–parasite dynamics that occur in marine species such as herrings Clupea spp., which have less well-defined population structures.

Alaska

Exxon Valdez oil spill restoration project final report: Prince William Sound Herring disease program (HDP), restoration project 070819

Surveys of pathogens in Pacific herring from 2007 – 2010 indicated that Ichthyophonus , viral hemorrhagic septicemia virus, and erythrocytic necrosis virus are endemic in Prince William Sound and throughout the NE Pacific. Laboratory studies with VHSV indicated that multiple herring stocks are equally susceptible to the resulting disease, Pacific herring shed copious levels of VHSV (as high at 5x10 8 plaque-forming units/day) shortly after exposure, chronic and persistent infections can occur in Pacific herring, susceptibility of Pacific herring to VHS extends to the larval life stages but not the embryonic stages, and the prior exposure history of Pacific herring to VHSV can be determined post hoc. Laboratory studies involving Ichthyophonus indicated that schizonts can be inactivated with chlorine and iodine solutions, the parasite can survive for extended periods in saltwater but not freshwater, a low potential exists for cross contamination between in vitro explant cultures, infectious schizonts are released from the skin surface of infected herring, schizonts are not uniformly distributed throughout the skeletal muscle of infected Pacific herring, multiple types of Ichthyophonus exist with different genotypes and phenotypic traits, and temperature is an important factor influencing the infectivity of Ichthyophonus . Additional field and laboratory studies indicated that Ichthyophonus negatively influences the swimming performance of infected hosts and the negative impacts effects are exacerbated by increasing temperatures, American shad are an important reservoir of Ichthyophonus in the NE Pacific, Pacific herring are not susceptible to infectious hematopoietic necrosis (IHN), and Pacific herring will accept surgically implanted acoustic tags with negligible impacts on survival.

Alaska, Washington

Effects of temperature on disease progression and swimming stamina in Ichthyophonus‐infected rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss , were infected with Ichthyophonus sp. and held at 10°C, 15°C and 20°C for 28 days to monitor mortality and disease progression. Infected fish demonstrated more rapid onset of disease, higher parasite load, more severe host tissue reaction and reduced mean‐day‐to‐death at higher temperature. In a second experiment, Ichthyophonus ‐infected fish were reared at 15°C for 16 weeks then subjected to forced swimming at 10°C, 15°C and 20°C. Stamina improved significantly with increased temperature in uninfected fish; however, this was not observed for infected fish. The difference in performance between infected and uninfected fish became significant at 15°C ( P =0.02) and highly significant at 20°C ( P = 0.005). These results have implications for changes in the ecology of fish diseases in the face of global warming and demonstrate the effects of higher temperature on the progression and severity of ichthyophoniasis as well as on swimming stamina, a critical fitness trait of salmonids. This study helps explain field observations showing the recent emergence of clinical ichthyophoniasis in Yukon River Chinook salmon later in their spawning migration when water temperatures were high, as well as the apparent failure of a substantial percentage of infected fish to successfully reach their natal spawning areas.

Journal of Fish Diseases

Isolation and characterization of a rhabdovirus from starry flounder (Platichthys stellatus) collected from the northern portion of Puget Sound, Washington, USA

The initial characterization of a rhabdovirus isolated from a single, asymptomatic starry flounder (Platichthys stellatus) collected during a viral survey of marine fishes from the northern portion of Puget Sound, Washington, USA, is reported. Virions were bullet-shaped and approximately 100 nm long and 50 nm wide, contained a lipid envelope, remained stable for at least 14 days at temperatures ranging from -80 to 5 degrees C and grew optimally at 15 degrees C in cultures of epithelioma papulosum cyprini (EPC) cells. The cytopathic effect on EPC cell monolayers was characterized by raised foci containing rounded masses of cells. Pyknotic and dark-staining nuclei that also showed signs of karyorrhexis were observed following haematoxylin and eosin, May-Grunwald Giemsa and acridine orange staining. PAGE of the structural proteins and PCR assays using primers specific for other known fish rhabdoviruses, including Infectious hematopoietic necrosis virus, Viral hemorrhagic septicemia virus, Spring viremia of carp virus, and Hirame rhabdovirus, indicated that the new virus, tentatively termed starry flounder rhabdovirus (SFRV), was previously undescribed in marine fishes from this region. In addition, sequence analysis of 2678 nt of the amino portion of the viral polymerase gene indicated that SFRV was genetically distinct from other members of the family Rhabdoviridae for which sequence data are available. Detection of this virus during a limited viral survey of wild fishes emphasizes the void of knowledge regarding the diversity of viruses that naturally infect marine fish species in the North Pacific Ocean.

Journal of General Virology

Duck plague virus replication in Muscovy duck fibroblast cells

Duck embryo fibroblast cell cultures from seven species of ducks were compared for virus yield, plaque quality, and sensitivity to infection by the duck plague herpesvirus (duck virus enteritis). Muscovy duck and wood duck cells gave the best results for virus yield and plaque quality, but muscovies were considered superior because they are more available than wood ducks. Pintails and lesser scaup gave the poorest results, and pekin duck, black duck, and redhead duck were intermediate. A growth curve for the virus, determined in muscovy cells, had a latent period of six hours and a maximum new virus titer reached at 36 hours. Because of their superior plaque production and ability to replicate the virus, muscovy embryo fibroblasts (MCE) are recommended for diagnostic and research work.

Avian Diseases