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

The estuarine genus Pfiesteria has received considerable attention since it was first identified and proposed to be the causative agent of fish kills along the mid-Atlantic coast in 1992. The presumption has been that the mechanism of fish death is by release of one or more toxins by the dinoflagellate. In this report, we challenge the notion that Pfiesteria species produce ichthyotoxins. Specifically, we show that (i) simple centrifugation, with and without ultrasonication, is sufficient to "detoxify" water of actively fish-killing cultures of Pfiesteria shumwayae, (ii) organic extracts of lyophilized cultures are not toxic to fish, (iii) degenerate primers that amplify PKS genes from several polyketide-producing dinoflagellates failed to yield a product with P. shumwayae DNA or cDNA, and (iv) degenerate primers for NRPS genes failed to amplify any NRPS genes but (unexpectedly) yielded a band (among several) that corresponded to known or putative PKSs and fatty acid synthases. We conclude that P. shumwayae is able to kill fish by means other than releasing a toxin into bulk water. Alternative explanations of the effects attributed to Pfiesteria are suggested.

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

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.

York river destratification: an estuary-subestuary interaction.

Destratification in the York River during high spring tides is the result of the interruption of normal two-layer estuarine flow by the advection of relatively fresh water into the river mouth from the Chesapeake Bay. This advection is due to the presence of a longitudinal salinity gradient in the bay and a difference in the tidal current phase between the river and the bay. Similar behavior is seen in other subestuaries of the Chesapeake Bay and may be common in subestuary-estuary interactions.

Ultrastructure of a marine Synechococcus possessing spinae.

Two Chesapeake Bay isolates of unicellular cyanobacteria belonging to the genus Synechococcus are described. Unicellular cyanobacteria are suspected to be important primary producers in estuarine and marine waters. One isolate (P-11-16) fluoresces red and forms green colonies. The other isolate (P-11-17) fluoresces orange and forms red colonies. Their ultrastructure is very similar to other isolates of Synechococcus except that spinae are formed and are attached to an outer wall layer not found in previously described species. The spinae are straight-walled cylinders, not flared at the base, are 44.0-65.0 nm in diameter, and range up to 2.7 micrometer in length. Substructure of the spinae wall consists of either material organized into stacks of rings or a strand of material helically coiled at a low (1-6 degrees) angle. Such material yielded a 6.0-9.2 nm cross-banding periodicity. Substructure of the rings or strand appeared to consist of bar-shaped, repeating units as seen in negatively stained material. Other procaryotic cell types with spinae, which were isolated from unincubated, natural seawater, are described.