Etiology of ulcerative lesions of Atlantic menhaden (Brevoortia tyrannus) from James River, Virginia.

We observed ulcerative lesions on live Atlantic menhaden, Brevoortia tyrannus, during ichthyofaunal sampling in the tidal James River in October 1999 (near Jamestown, VA, USA). Other synoptically collected fishes exhibited no signs of lesions or pre-ulcerative tissues. Live fish were classified as unremarkable (no dermal anomalies), pre-ulcerative (integument intact with boil-like swelling), and ulcerative (severe focal lesions). Specimens were analyzed for bacteria, fungi, and pathogenic protozoans including amphizoic amoebae, Pfiesteria piscicida, and Kudoa sp. No Pfiesteria were detected in any tissue specimen. All B. tyrannus examined, including tissues from unremarkable fish, tested positive for presence of the known fish parasite Kudoa. Only ulcerative lesions were also colonized by bacteria, fungi, and amphizoic amoebae. The absence of bacteria, fungi, and protozoans from unremarkable and pre-ulcerative fish suggests that association of other potential pathogens with B. tyrannus ulcers was due to secondary colonization following lesion formation as a result of Kudoa infection.

Detection of Bacillus endospores using total luminescence spectroscopy.

Detection and analysis of bacteria from environmental samples (e.g. water, air, and food) are usually accomplished by standard culture techniques or by analyses that target specific DNA sequences, antigens or chemicals. For large cell numbers in aqueous suspensions, an alternative technique that has proven useful is total luminescence spectroscopy (TLS). TLS is the acquisition of fluorescence data that records the unique excitation-emission matrix (EEM) of compound fluorophores. Past work has shown that one type of bacterial endospore, Bacillus megaterium, possessed a distinct EEM pattern useful for differentiating it in complex biological fluids and suspensions. The work described here extends those observations to establish some limits on the sensitivity and specificity of TLS for the detection and analysis of bacterial endospores versus (bacterial) vegetative cells in aqueous culture. Our findings show Bacillus endospores exhibit a dramatic blue shift of 130 nm in excitation and a smaller shift of 50 nm in emission when compared to ancillary endospore and non-endospore forming bacterial cells.

Distribution of free-living amoebae in James River, Virginia, USA.

A comprehensive survey to document the presence of free-living amoebae was conducted along 58 km of James River, near Richmond, Virginia, USA. Sites included tidal and non-tidal freshwater areas, near 40 combined sewer outflows, three municipal wastewater treatment plant release sites, and thermal discharge from a coal-fired power plant. Amoebae were present on all collection dates, spring through autumn, and at all sites ( n=330). Five genera, Naegleria, Vannella, Acanthamoeba, Vahlkampfia, and Hartmannella were present in both the water column and sediment. The most common isolates from the water column were Naegleria and Vannella. Water conditions conducive to the presence of large quantities of fecal coliform bacteria were correlated with the prevalence of free-living amoebae. Some of the amoebae in this complex ecosystem can act as opportunistic pathogens, may play a role in diseases of aquatic organisms in this heavily urbanized river, and may present a risk to human health.