Skin ulcers in estuarine fishes: a comparative pathological evaluation of wild and laboratory-exposed fish.

Abstract

The toxic dinoflagellate Pfiesteria piscicida Steidinger & Burkholder has recently been implicated as the etiologic agent of acute mass mortalities and skin ulcers in menhaden, Brevoortia tyrannus, and other fishes from mid-Atlantic U.S. estuaries. However, evidence for this association is largely circumstantial and controversial. We exposed tilapia (Oreochromis spp.) to Pfiesteria shumwayae Glasgow & Burkholder (identification based on scanning electron microscopy and molecular analyses) and compared the resulting pathology to the so-called Pfiesteria-specific lesions occurring in wild menhaden. The tilapia challenged by high concentrations (2,000-12,000 cells/mL) of P. shumwayaeexhibited loss of mucus coat and scales plus mild petecchial hemorrhage, but no deeply penetrating chronic ulcers like those in wild menhaden. Histologically, fish exhibited epidermal erosion with bacterial colonization but minimal associated inflammation. In moribund fish, loss of epidermis was widespread over large portions of the body. Similar erosion occurred in the mucosa lining the oral and branchial cavities. Gills exhibited epithelial lifting, loss of secondary lamellar structure, and infiltration by lymphoid cells. Epithelial lining of the lateral line canal (LLC) and olfactory organs exhibited severe necrosis. Visceral organs, kidney, and neural tissues (brain, spinal cord, ganglia, peripheral nerves) were histologically normal. An unexpected finding was the numerous P. shumwayae cells adhering to damaged skin, skin folds, scale pockets, LLC, and olfactory tissues. In contrast, histologic evaluation of skin ulcers in over 200 wild menhaden from Virginia and Maryland portions of the Chesapeake Bay and the Pamlico Estuary, North Carolina, revealed that all ulcers harbored a deeply invasive, highly pathogenic fungus now known to be Aphanomyces invadans. In menhaden the infection always elicited severe myonecrosis and intense granulomatous myositis. The consistent occurrence of this fungus and the nature and severity of the resulting inflammatory response indicate that these ulcers are chronic (age >1 week) and of an infectious etiology, not the direct result of an acute toxicosis initiated by Pfiesteria toxin(s) as recently hypothesized. The disease therefore is best called ulcerative mycosis (UM). This study indicates that the pathology of Pfiesteria laboratory exposure is fundamentally different from that of UM in menhaden; however, we cannot rule out Pfiesteria as one of many possible early initiators predisposing wild fishes to fungal infection in some circumstances.

Characterization of Ichthyocidal Activity of Pfiesteria piscicida : Dependence on the Dinospore Cell Density

“…The evidence presented here also supports reexamination of the rationale for the interpretation of results obtained in cell receptor assays using bioassay supernatants, as well as environmental water (15). Our results suggest that, as described for P. shumwayae (43), any contribution of Pfiesteria spp. to fish death in the bioassay system would be mostly mediated by direct interactions of the dinospore with fish external surfaces, as a result of their feeding behavior.…”

Molecular Assays for DetectingAphanomyces invadansin Ulcerative Mycotic Fish Lesions

“…The relationship between Pfiesteria and fish lesions, however, was based on laboratory studies that did not simulate natural conditions. Those studies were challenged by a number of researchers (10,11,(41)(42)(43)(44), and there is now a large body of research evidence that supports the primary role of A. invadans in causing UM. Repeated histological surveys of UM-affected fish have found a high number of broad aseptate hyphae in the ulcers (2,4,12,13,24,25,35).…”

Are Pfiesteria species toxicogenic? Evidence against production of ichthyotoxins by Pfiesteria shumwayae

“…A clonal isolate of Pfiesteria shumwayae [Center for Culture of Marine Phytoplankton (CCMP) 2089] was maintained in cultures as described below. The identity of P. shumwayae was confirmed additionally by PCR using species-specific primers for regions of the ribosomal RNA gene complex (29).…”