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Featuring       the latest information on ground-water and surface-water in the MD-DE-DC       district.**SEARCH         THE DISTRICT:****National       District Sites**Choose           BelowAlabamaAlaskaArizonaArkansasCaliforniaColoradoConnecticutDelawareFloridaGeorgiaHawaiiIdahoIllinoisIndianaIowaKansasMaineMassachusettsMichiganMinnesotaMississippiMissouriMontanaNebraskaNevadaNew HampshireNew JerseyNew MexicoNew YorkNorth CarolinaNorth DakotaOhioOklahomaOregonPennsylvaniaPuerto RicoRhode IslandSouth CarolinaSouth DakotaTennesseeTexasUtahVermontVirginiaWashingtonWest VirginiaWisconsinWyoming________________****Major       Cooperators**** ****Online                           Publication - FS-114-98****** Home                           >>District                           Publications                           >> Online Publication FS-114-98**Fish                                 Health, Fungal Infections, and *Pfiesteria*:                                The Role of the U.S. Geological Survey** by                                 Vicki Blazer, Scott Phillips, and Edward Pendleton                                 
**Problem**                                               The                                         increasing incidence of fish-health problems,                                         ranging from small external sores and                                         lesions to large-scale fish kills in the                                         Chesapeake Bay and other Mid-Atlantic                                         estuaries, is of growing concern to natural                                         resource managers and residents of the                                         Chesapeake Bay watershed. Fish kills have                                         been reported in North Carolina since                                         the early 1990's. Reports of skin lesions                                         on fish in the summer and fall of 1996                                         and 1997 in the Pocomoke River, Maryland                                         and other tributaries of Chesapeake Bay                                         stimulated a great deal of public and                                         scientific interest. These skin lesions                                         ranged from small pinpoint hemorrhages                                         to abrasions to deep ulcers. In addition,                                         there were two fish kills, involving primarily                                         Atlantic menhaden, in the Pocomoke River                                         during August 1997. The fish kills, as                                         well as the variety of fish lesions, have                                         been blamed on the presence of a toxic                                         dinoflagellate, *Pfiesteria piscicida*,                                         or *Pfiesteria*-like dinoflagellates.                                         The occurrence of *Pfiesteria* has                                         been attributed to nutrient enrichment,                                         warm water temperatures, moderate to high                                         salinity, and fish excretions (Burkholder                                         and Glasgow, 1997).                                               While                                         *Pfiesteria* has been identified                                         as a potential cause of fish kills in                                         the Chesapeake Bay and other estuaries,                                         there is growing evidence that other pathogens                                         also are causing lesions in fish in the                                         Bay. Skin lesions in fish can be caused                                         by a variety of infectious and noninfectious                                         insults. Lesions commonly are caused by                                         opportunistic pathogens, which infect                                         weakened or stressed hosts. These pathogens,                                         which include bacteria, fungi, viruses,                                         and parasites, may gain entry because                                         of impaired immune or disease resistance                                         factors, or because the natural defense                                         mechanisms of the skin are impaired or                                         breached.                                               Several                                         State and Federal agencies, including                                         the U.S. Geological Survey (USGS), are                                         currently working to understand the relation                                         between *Pfiesteria* and other pathogens                                         to fish and human health, and to natural                                         and man-induced factors (such as nutrients                                         and sediment). This information will be                                         used to help formulate management solutions                                         to the problem. 

**USGS                                         investigations of fish lesions, related                                         environmental factors, and nutrient sources**                                               In                                         1983, the U.S. Environmental Protection                                         Agency began the Chesapeake Bay Program                                         with other Federal Agencies and several                                         States to restore the ecosystem of the                                         Bay and its watershed. The USGS has been                                         an active partner since the formation                                         of this program, and has provided scientific                                         information on the relation between contaminant                                         inputs (nutrients, sediment, and toxins)                                         and the response of water quality and                                         living resources.                                                In                                         1996, the USGS Chesapeake Bay Ecosystem                                         Program was initiated to provide scientific                                         information to resource managers on the                                         response of water quality and living resources                                         to changes in nutrient sources. As part                                         of the Ecosystem and other USGS Programs,                                         the USGS Leetown Science Center and the                                         Maryland Cooperative Fish and Wildlife                                         Research Unit began investigations to                                         determine factors contributing to fish                                         lesions in selected tributaries of the                                         Chesapeake Bay. USGS studies include characterizing                                         the type and cause of fish lesions and                                         evaluating overall fish health using a                                         variety of physiological, histological,                                         and immunological techniques. Additionally,                                         researchers at Leetown work with other                                         USGS scientists to document the potential                                         linkages between the occurrence of lesions                                         and water quality, land use, and nutrient                                         sources and transport to the Bay. Some                                         of the locations of different components                                         of USGS studies of fish health and nutrient                                         sources and transport are shown in figure                                         1. 
                                             Figure 1. Location of USGS fish collection,                                             streamflow, and water-quality sampling                                             sites near *Pfiesteria* outbreaks                                             on Chesapeake Bay tributaries in Maryland                                             and Delaware. 
**The                                         cause of fish lesions: fungal infections                                         or *Pfiesteria*?**                                               The                                         USGS and State partners collected menhaden                                         from sites affected by fish kills and                                         river closures from August through October                                         1997. Menhaden with lesions also were                                         collected in the Wicomico and Pocomoke                                         Rivers in August 1998 (Fig. 1). Ninety-five                                         percent of the menhaden with lesions collected                                         in 1997 and 100 percent collected in 1998                                         had chronic sores in response to an invasive                                         fungus (Fig. 2). 
                                             Figure 2. Lesions of menhaden that                                             contain the fungal pathogen: ***A***                                             early raised lesion; ***B***                                             advanced ulcerated lesion.                                              Figure 3. ***A*** Section                                             of lesion skeletal muscle of menhaden;                                             most of the normal muscle is replaced                                             with chronic inflammation surrounding                                             invading fungus (thin arrows); remnants                                             of normal muscle bundles (thick arrows)                                             stain pink-red with routine histologic                                             stain, hematocrit and eosin (H&E);                                             inflammatory cells appear purplish-blue                                             (240x). ***B*** Section of                                             normal skeletal muscle of menhaden                                             arranged in bundles and stained pink-red                                             with routine histologic (H&E)                                             stain (240x). 
                                               The                                         stained microscopic appearance (Fig. 3A)                                         of most of these lesions is identical                                         to that of lesions observed on fish in                                         other estuarine environments where cutaneous                                         ulcerative syndromes are associated with                                         an invasive fungus called *Aphanomyces                                         invadans* (Fig. 4). These syndromes                                         include red spot disease (RSD) in Australia,                                         epizootic ulcerative syndrome (EUS) of                                         Asian fishes, and mycotic granulomatosis                                         (MG) in Japan. Outbreaks of EUS and RSD                                         have been correlated with rainfall-runoff                                         events, warm water temperatures, and low                                         levels of dissolved oxygen (less than                                         one milligram per liter).                                               Previous                                         research activities suggest that the fungal                                         infections seen in menhaden and other                                         estuarine fish in North Carolina are secondary                                         to exposure of the fish to toxins from                                         *Pfiesteria* (Noga and others, 1996;                                         Burkholder and Glasgow, 1997). But scientific                                         evidence of a relation between these lesions                                         and exposure to toxic dinoflagellates                                         is lacking.                                                In                                         1998, a fungus was isolated from lesioned                                         menhaden in the Wicomico and Pocomoke                                         Rivers. By preliminary analyses including                                         growth requirements, growth-temperature                                         curves and appearance in culture these                                         isolates appear to be similar or identical                                         to *Aphanomyces invadans* (James                                         Lilley, oral commun., 1998, Stirling University,                                         Stirling, Scotland).                                                Laboratory                                         exposures to *Pfiesteria* have not                                         resulted in similar lesions. Respiratory                                         distress, neurological dysfunction, and                                         a cloudiness of the skin with acute mortalities                                         were reported in early laboratory studies                                         (Smith and others, 1988; Noga and others,                                         1993).                                               Microscopic                                         study of skin shows swelling of cells                                         and the death of skin cells, resulting                                         in deep sores extending through the skin                                         (Noga and others, 1996). This study also                                         examined fish that survived an acute sublethal                                         exposure to *Pfiesteria piscicida*                                         and then were placed into clean water.                                         One of ten fish developed a fungal infection.                                         This infection, however, appeared typical                                         of fungal organisms growing in necrotic                                         tissue. The fungus did not cause the typical                                         granulomatous response (Fig. 3A) seen                                         in wild fish captured in Bay tributaries.                                                                                        Burkholder                                         and Glasgow (1997) stated that "the fungi                                         are opportunists; their hyphae generally                                         do not penetrate to the lesion base, suggesting                                         they do not form the lesions; rather,                                         they colonize lesions formed by toxin(s)                                         from *Pfiesteria piscicida*." This                                         phenomenon was not observed in menhaden                                         collected in the Chesapeake tributaries.                                         In many instances, the fungal organism                                         and surrounding chronic inflammation extended                                         deep into muscle, under intact, normal                                         skin. In a number of menhaden, the fungal                                         organism had actually penetrated to and                                         through visceral organs such as the liver.                                         
                                             Figure 4. Fungal structures, darkly                                             stained by Grocott, are visible within                                             muscle tissue of menhaden. 
**Preliminary                                         information on nutrient sources and environmental                                         factors affecting fish health**                                               The                                         USGS is compiling information on estuarine                                         water quality and the human and landscape                                         factors that may affect fish health, such                                         as road density, population density, forest                                         fragmentation, and patterns of nutrient                                         sources. Additionally, information on                                         land characteristics that influence nutrient                                         transport, such as soil type, riparian                                         zones, extent of drainage ditching, and                                         geology is being compiled and integrated                                         with streamflow and water-quality data                                         being collected in the Pocomoke River                                         Basin.                                                Resource                                         managers will use the information on nutrient                                         sources and transport to evaluate management                                         practices that are being implemented to                                         reduce nutrients entering the Bay and                                         tributaries. In Maryland, legislation                                         has been passed requiring farmers to adopt                                         plans to control nitrogen runoff by July                                         2001 and phosphorous runoff by July 2004.                                                                                        Excess                                         nutrients (phosphorus and nitrogen) in                                         the soil can either run off into streams                                         directly or percolate through the soil                                         to the water table and reach streams as                                         ground-water discharge. Phosphorus tends                                         to bind with sediment particles and is                                         transported primarily with surface water,                                         whereas dissolved nitrogen travels in                                         both surface water and ground water. Ground                                         water in shallow aquifers that discharges                                         to streams is called "base flow."                                                The                                         USGS has found that the proportion of                                         streamflow contributed by base flow in                                         shallow aquifers of the Coastal Plain                                         ranges from approximately 42 to 74 percent                                         (Bachman and others, 1998). On the basis                                         of existing data, the concentration of                                         nitrate in shallow aquifers underlying                                         the lower Eastern Shore of Maryland generally                                         ranges from less than 0.1 to 35 milligrams                                         per liter. The associated age of most                                         of the water in shallow Coastal Plain                                         aquifers ranges from 0 to 20 years (Focazio                                         and others, 1998). This indicates that,                                         despite recent changes in management practices                                         to control nitrogen, an improvement in                                         surface-water quality may take a number                                         of years to evolve. The USGS is currently                                         collecting additional information to better                                         document the nutrients and associated                                         ground-water residence times in aquifers                                         discharging to surface-water bodies in                                         the Pocomoke River Basin. 

**Future                                         issues and management implications**                                               *Pfiesteria*                                         was detected in Maryland waters in 1997                                         and was believed to be the primary cause                                         of fish kills. Maryland has established                                         criteria to close rivers on the basis                                         of both fish kills, fish lesions, and                                         other factors. Therefore, there is a need                                         for resource managers to understand the                                         causes of both fish kills and lesions.                                         While it is indeed possible that toxic                                         dinoflagellates may play a role in the                                         initiation of fish lesions, other factors                                         also need to be considered. The finding                                         of massive cell death over top of the                                         chronic tissue damage in the muscle of                                         fish from the river closure areas suggests                                         these inflamed lesions were present prior                                         to exposure to *Pfiesteria* toxin.                                         It is possible that large numbers of migrating                                         menhaden with open skin sores could stimulate                                         the dinoflagellate blooms. Further research                                         is needed to study the development of                                         this fungal disease in menhaden, to understand                                         the underlying stressors that may initiate                                         this and other infections either by skin                                         damage or immunosuppression, and to understand                                         what role toxic dinoflagellates play in                                         these lesions. It is also important to                                         determine the time of year when the menhaden                                         first become infected and to follow the                                         development of these chronic lesions.                                         In addition to the occurrence of lesions,                                         the environmental factors -- nutrients,                                         contaminants, fresh-water flow, and estuarine                                         conditions-contributing to their occurrence                                         must be understood in order to formulate                                         effective management solutions. The USGS                                         continues to work with appropriate State                                         and Federal agencies to address fish health                                         issues in the Chesapeake Bay. 

**References**
-  Bachman,                                             L.J., Lindsey, B.D., Brakebill, J.B.,                                             and Powars, D.S., 1998, Ground-water                                             discharge and base-flow nitrate loads                                             of nontidal streams, and their relation                                             to a hydrogeomorphic classification                                             of the Chesapeake Bay watershed, Middle                                             Atlantic Coast: U.S. Geological Survey                                             Water-Resources Investigations Report                                             98-4059, 7l p.  
-  Burkholder,                                             J.M., and Glasgow, H.B., Jr., 1997,                                             *Pfiesteria piscicida* and other                                             *Pfiesteria*-like dinoflagellates:                                             Behavior, impacts, and environmental                                             controls: Limnol. Oceanogra. V. 42,                                             p. 1052-1075.  
-  Focazio,                                             M.J., Plummer, L.N., Bohlke, J.K.,                                             Busenberg, Eurybiades, Bachman, L.J.,                                             and Powars, D.S., 1998, Preliminary                                             estimates of residence times and apparent                                             ages of ground water in the Chesapeake                                             Bay watershed, and water-quality data                                             from a survey of springs: U.S. Geological                                             Survey Water-Resources Investigations                                             Report 97-4225, 75 p.  
-  Noga,                                             E.J., Smith, S.A., Burkholder, J.M.,                                             Hobbs, C., and Bullis, R.A., 1993,                                             A new ichthyotoxic dinoflagellate:                                             cause of acute mortality in aquarium                                             fishes: Vet Rec., V. 133, p. 96-97.                                              
-  Noga,                                             E.J., Khoo, J.B., Stevens, Z. F.,                                             and Burkholder, J.M., 1996, Novel                                             toxic dinoflagellate causes epidemic                                             disease in estuarine fish: Marine                                             Pollution Bulletin V. 32, p. 219-224.                                              
-  Smith,                                             S.A., Noga, E.J., and Bullis, R.A.,                                             1988, Mortality in *Tilapia aurea*                                             due to a toxic dinoflagellate bloom:                                             Abstract *in* Proceedings of                                             the Third International Colloq. Pathologic                                             Marine Aquaculture: Gloucester Point,                                             Virginia, p. 167-168.  

For                                           further information contact: 

                                         Chesapeake Bay Program Coordinator                                        U.S. Geological Survey                                        8987 Yellow Brick Road                                        Baltimore, Maryland 21237 

                                         or visit the USGS Chesapeake Bay Homepage:http://chesapeake.usgs.gov/chesbay/                                         
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