Jan Lovy

Parvicapsula minibicornis infections in gill and kidney and the premature mortality of adult sockeye salmon (Oncorhynchus nerka) from Cultus Lake, British Columbia

In recent years, large losses of migrating adult sockeye salmon (Oncorhynchus nerka) from the Fraser River, British Columbia, Canada, have resulted in reductions in catches and productivity. We investigated patterns of mortality and the occurrence of the myxosporean parasite Parvicapsula minibicornis in adult sockeye salmon from Cultus Lake, tributary to the Fraser River. Using data from a captive broodstock program, we found that early migrants to Cultus Lake had less severe P. minibicornis infections and were more likely to survive to maturity than those fish that arrived later after they had presumably held in the warmer Fraser River. We found P. minibicornis in kidneys and gills of spawners. In some fish, significant histopathology in the gills that included severe inflammation and hyperplasia of the gill lamellae was observed; the severity of the disease was correlated with the severity of P. minibicornis infections. Kidney and gill pathologies were more prevalent and more severe in fish that died be...

Molecular epidemiology of viral haemorrhagic septicaemia virus (VHSV) in British Columbia, Canada, reveals transmission from wild to farmed fish

Viral haemorrhagic septicaemia virus (VHSV) is a fish pathogen found throughout the Northern Hemisphere and is capable of infecting and causing mortality in numerous marine and freshwater hosts. In the coastal waters of British Columbia, Canada, the virus has been detected for 20 yr with many occurrences of mass mortalities among populations of Pacific herring Clupea pallasii (Valenciennes) and sardine Sardinops sagax as well as detections among cultured Atlantic Salmo salar and Chinook Oncorhynchus tshawytscha salmon. We compared nucleotide sequence of the full glycoprotein (G) gene coding region (1524 nt) of 63 VHSV isolates sampled during its recorded presence from 1993 to 2011 from 6 species and a total of 29 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVa within the major VHSV genetic group IV. Of the 63 virus isolates, there were 42 unique sequences, each of which was ephemeral, being repeatedly detected at most only 1 yr after its initial detection. Multiple sequence types were revealed during single viral outbreak events, and genetic heterogeneity was observed within isolates from individual fish. Moreover, phylogenetic analysis revealed a close genetic linkage between VHSV isolates obtained from pelagic finfish species and farmed salmonids, providing evidence for virus transmission from wild to farmed fish.

Comparative Cellular Morphology Suggesting the Existence of Resident Dendritic Cells Within Immune Organs of Salmonids

This report is the first morphological description of cells that resemble dendritic cells, which appear to form resident populations within the spleen and anterior kidney of fish. Based on examination of three salmonid species, including, rainbow trout, brook trout, and Atlantic salmon, the cells were most abundant in the spleen, although they were always present in the anterior kidney. The cells appeared diffusely distributed, often near blood vessels of the spleen and kidney of healthy fish and within the epithelium, connective tissue, and blood vessels of rainbow trout gills with experimentally induced microsoporidial gill disease. The dendritic‐like cells in this study contained granules that resemble Birbeck granules, which are considered to be morphological markers of Langerhans cells in mammals. The cells were approximately 6 μm in diameter and contained Birbeck‐like (BL) granules localized near centrioles. Although the dendritic‐like cells in the three salmonid species shared many similarities, morphological differences were found in the fine structure of the rod portion of the BL granules. Rainbow trout BL granules contained amorphous material, while the other salmonid species contained particulate material arranged in a square‐lattice arrangement. The BL granules in the cells of Atlantic salmon had a narrow diameter and contained four layers of particulate material when sectioned longitudinally; two layers enveloped by the granule membrane and two central layers making up a central lamella, which is common in mammalian Birbeck granules. Anat Rec, 291:456–462, 2008.

Isolation of Edwardsiella ictaluri from Tadpole Madtom in a Southwestern New Jersey River

Abstract In 2002, New Jersey Division of Fish and Wildlife biologists conducted an aquatic assessment in a section of the Cohansey River in Cumberland County. Three of seven tadpole madtoms Noturus gyrinus collected were observed to have an ulcerative lesion on the head between the eyes. Gram-negative, oxidase-negative rods were isolated from the head kidneys of two of the madtoms and from the cranial ulcer of the third madtom. Four different identification methods— API-20E, whole-cell fatty acid methyl esters (FAME), BIOLOG, and enterobacterial repetitive intergenic consensus (ERIC) sequences—ascribed the isolates to the species Edwardsiella ictaluri. A phenotypic comparison using FAME and BIOLOG data revealed that E. ictaluri isolates from wild madtoms shared a high degree of similarity to E. ictaluri isolates from cultured channel catfish with enteric septicemia of catfish (ESC) from southeastern states. However, fingerprinting by amplified fragment length polymorphisms (AFLP) demonstrated that the thr...

Infection of gill and kidney of Fraser River sockeye salmon, Oncorhynchus nerka (Walbaum), by Parvicapsula minibicornis and its effect on host physiology

Adult sockeye salmon, Oncorhynchus nerka (Walbaum), migrating upstream in the Fraser River, British Columbia, are exposed to the myxozoan parasite Parvicapsula minibicornis when they enter the river from the ocean. Infections are initially localized in the kidney but have recently been associated with branchitis in one population. Adult fish from five locations in the watershed were sampled to determine whether branchitis was widespread. P. minibicornis infections in kidney glomeruli were prevalent in all samples except for a sample of fish that had just entered the Fraser River from the ocean. For fish captured in spawning streams, parasites were observed in the renal tubules and gill, and branchitis was observed in 70% of fish. Plasma osmolality was negatively correlated with the number of parasites in the kidney tubules, which we hypothesize to be caused by the breach of glomerular membranes as the parasite leaves the fish. Plasma lactate values increased with increasing levels of pathology in gills. These findings support the hypothesis that P. minibicornis impacts the physiology of migrating fish, which may in turn affect the likelihood that adults will be able to migrate and spawn successfully.

Effects of dexamethasone on host innate and adaptive immune responses and parasite development in rainbow trout Oncorhynchus mykiss infected with Loma salmonae.

The effects of dexamethasone (dex) treatment on infections with the microsporidian parasite, Loma salmonae and the effects of dex on initiation of the adaptive immune response were investigated in rainbow trout, Oncorhynchus mykiss experimentally infected with the parasite. Dex treatment resulted in significantly higher infections with the parasite in the gills and other internal organs, suggesting that dex inhibits aspects of the innate immune response to L. salmonae; the heavier infections in the gills and organs of rainbow trout resembled infections seen in Chinook salmon. Mean xenoma counts per microscope field in the gills of fish infected with L. salmonae treated with dex or left untreated were 169 and 30, respectively. Although higher numbers of xenomas were observed in dex treated fish, the xenomas were generally smaller in size than in infected control fish. The xenomas in dex treated fish showed morphological signs of degeneration including loss and degeneration of early parasite stages, accumulation of amorphous material in xenomas, and infiltration with phagocytic cells containing degenerated parasites. The xenomas in infected untreated fish had larger xenomas with a more uniform size and contained identifiable parasite stages in the cytoplasm. According to this study, once fish have developed an adaptive immune response to the parasite by previous exposure, then fish have 100% protection to reinfection even when treated with heavy doses of dex. L. salmonae immune fish treated or untreated with dex during reinfection with the parasite developed no xenomas in the gills 6 weeks post reinfection. These results indicate that once the cellular response is primed to L. salmonae, then dex related immunosuppression does not reduce the effectiveness of the adaptive immune response.

The parasite Ichthyophonus sp. in Pacific herring from the coastal NE Pacific

The protistan parasite Ichthyophonus occurred in populations of Pacific herring Clupea pallasii Valenciennes throughout coastal areas of the NE Pacific, ranging from Puget Sound, WA north to the Gulf of Alaska, AK. Infection prevalence in local Pacific herring stocks varied seasonally and annually, and a general pattern of increasing prevalence with host size and/or age persisted throughout the NE Pacific. An exception to this zoographic pattern occurred among a group of juvenile, age 1+ year Pacific herring from Cordova Harbor, AK in June 2010, which demonstrated an unusually high infection prevalence of 35%. Reasons for this anomaly were hypothesized to involve anthropogenic influences that resulted in locally elevated infection pressures. Interannual declines in infection prevalence from some populations (e.g. Lower Cook Inlet, AK; from 20-32% in 2007 to 0-3% during 2009-13) or from the largest size cohorts of other populations (e.g. Sitka Sound, AK; from 62.5% in 2007 to 19.6% in 2013) were likely a reflection of selective mortality among the infected cohorts. All available information for Ichthyophonus in the NE Pacific, including broad geographic range, low host specificity and presence in archived Pacific herring tissue samples dating to the 1980s, indicate a long-standing host-pathogen relationship.

Histochemical and ultrastructural analysis of pathology and cell responses in gills of channel catfish affected with proliferative gill disease.

Pond-reared channel catfish Ictalurus punctatus with proliferative gill disease (PGD), caused by the myxozoan parasite Henneguya spp., were examined with light and transmission electron microscopy to better characterize the inflammatory response during infection. The early stages of disease are characterized by the destruction of collagen in the matrix of the gill filament cartilage causing weakness and breaks within the gill filaments. These early lesions lacked a notable inflammatory response around the disrupted cartilage, a chondrocyte response was not apparent, and the parasite was not present, suggesting that the cartilage breaks occur prior to inflammation and arrival of the parasite in the gill. In later lesions, a significant inflammatory response was generated in areas of disrupted cartilage, and the inflammatory infiltrate was composed of a mixed population of granulocytes including neutrophils and cells that resembled eosinophils. The majority of eosinophil-like cells demonstrated evidence of degranulation. Trophozoites of Henneguya spp. were surrounded by a uniform population of cells believed to be neutrophils. The granulocytes were infiltrated within the dense collagen layer of the gill filament cartilage and often appeared within chondrocyte lacunae in place of the chondrocyte. The gill lamellae adjacent to the lesions were fused and contained an inflammatory infiltrate containing granulocytes and cells with pericentriolar granules that resembled previous descriptions of Langerhans-like cells. These cells were abundant within damaged lamellar epithelium, but were only rarely found within the gill filament. Lesions that appeared to be recovering lacked the dense collagenous layer around the cartilage and contained hyperplastic and hypertrophic chondrocytes that formed a callus. Other chondrocytes in the lesions had ultrastructural features indicative of cell death.

Phylogeny and morphology of Glugea hertwigi from rainbow smelt Osmerus mordax found in Prince Edward Island, Canada

Infection of rainbow smelt Osmerus mordax with the microsporidian Glugea hertwigi was diagnosed for the first time in Prince Edward Island, Canada. The prevalence of infection was 24%, 45 infected out of 187 examined fish captured in February and March 2009. Both large and small xenomas of G. hertwigi observed within the submucosa of the gastrointestinal tract and along the mesentery of the host contained only mature spores. Advanced and degraded xenomas associated with host reaction were described using light and transmission electron microscopy. The first rDNA sequence of G. hertwigi prepared in the present study completed the set of sequences of Glugea spp. available for comparison. The high level of rDNA sequence identity between Glugea spp. suggests that these may be variants of a single species.

Comparative ultrastructure of Langerhans‐like cells in spleens of ray‐finned fishes (Actinopterygii)

We studied the morphology and occurrence of splenic Langerhans‐like (LL) cells in species representing 11 orders of ray‐finned fishes, Actinopterygii. LL cells were frequent in spleen tissue of species among Cypriniformes, Esociformes, Salmoniformes, and Pleuronectiformes. These cells contained granules which resembled Birbeck granules known to occur in mammalian Langerhans cells. The ultrastructure of LL cells in Northern pike, Esox lucius, and in Atlantic halibut, Hippoglossus hippoglossus were similar to those reported in salmonids. LL cells found in cyprinids shared some characteristics with the LL cells in other Actinopterygii species, although unique structures distinguished them from the latter. They contained dense bodies within the Birbeck‐like (BL) granules, a characteristic that was never observed in species outside the Cypriniformes. Two types of BL granules were characterized in cyprinid LL cells. The ultrastructure of BL granules across the species is discussed. LL cells in all Actinopterygii species demonstrated close contacts with nearby cells, characterized by adherens‐like junctions. Additionally, multivesicular bodies were present within the cytoplasm and large aggregates of exosomes were observed closely associated with the plasma membrane suggesting their release from the cells. These structures are discussed in relation to mammalian dendritic cells. Macrophages found in European perch, Perca fluviatilis, blue gourami, Trichogaster trichopterus, and Atlantic halibut, Hippoglossus hippoglossus contained lysosomes and residual bodies with structures resembling Birbeck granules. These granules and cells were clearly distinct from LL cells. J. Morphol. 2010.