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Fecal-coliform bacteria concentrations in streams of the Chattahoochee River National Recreation Area, Metropolitan Atlanta, Georgia, May–October 1994 and 1995

The Metropolitan Atlanta area has been undergoing a period of rapid growth and development. The population in the 10-county metropolitan area almost doubled from about 1.5 million people in 1970 to 2.9 million people in 1995 (Atlanta Regional Commission, written commun., 2000). Residential, commercial, and other urban land uses more than tripled during the same period (Frick and others, 1998). The Chattahoochee River is the most utilized water resource in Georgia. The rapid growth of Metropolitan Atlanta and its location downstream of the headwaters of the drainage basin make the Chattahoochee River a vital resource for drinking-water supplies, recreational opportunities, and wastewater assimilation. In 1978, the U.S. Congress declared the natural, scenic, recreation, and other values of 48 miles of the Chattahoochee River from Buford Dam to Peachtree Creek to be of special national significance. To preserve this reach of the Chattahoochee River, the U.S. Congress created the Chattahoochee River National Recreational Area (CRNRA), which includes the Chattahoochee River downstream from Buford Dam to the mouth of Peachtree Creek and a series of park areas adjacent to the river in northern Metropolitan Atlanta Even with this protection, waters of the Chattahoochee River and many of its tributaries in Metropolitan Atlanta did not meet water-quality standards set for designated uses during 1994 and 1995 (fig. 1 and table 1). Much of the degradation of water quality has been associated with areas undergoing rapid urban growth and sprawling suburban development. The resulting conversion of mostly forested land to urban land has multiple adverse effects on water quality. Degradation of water quality may be caused by a number of factors including an increase in nutrient concentrations, sediment and sedimentbound contaminant concentrations (e.g., metals and pesticides) (Frick and others, 1998), and fecal-coliform bacteria concentrations (Center for Watershed Protection, 1999). The presence of fecal-coliform bacteria in streams and rivers indicates that contamination by fecal material from human or animal sources has occurred and contact with these waters can result in exposure to pathogenic bacteria often associated with fecal contamination. During 1994 and 1995, elevated concentrations of fecal-coliform bacteria were the most common reason that the Chattahoochee River and tributaries did not meet their designated uses of drinking-water supply, recreation, and fishing. According to the Georgia Department of Natural Resources (1997), during 1994 and 1995, 67 of 77 stream reaches assessed in Metropolitan Atlanta did not meet or only partially met water-quality requirements for designated uses. Excessive concentrations of fecal-coliform bacteria were a contributing factor in 63 of the 67 streams that did not meet or only partially met designated uses. High concentrations of fecal-coliform bacteria have the potential to reduce the recreational value of the river and pose a continued threat, with unknown health risks, to humans that come in contact with the water while fishing, boating, rafting, wading, and swimming.

Georgia↗

Fecal coliform and Escherichia coli bacteria in the St. Croix National Scenic Riverway, Summer 1999

Fecal coliform and Escherichia coli ( E. coli ) concentrations were determined in the St. Croix National Scenic Riverway to assess whether pathogenic organisms pose a potential problem for recreational use. Samples were collected from May through September 1999 at 22 locations on the St. Croix and Namekagon Rivers. No concentrations exceeded water-quality criteria or standards set by the U.S. Environmental Protection Agency or the states of Minnesota and Wisconsin. Maximum fecal coliform and E. coli concentrations were measured in the St. Croix River at St. Croix Falls, Wisconsin. Median fecal coliform and E. coli concentrations were greater in the St. Croix River near Woodland Corner, Wisconsin, and at Marine on St. Croix, Minnesota than at other locations sampled. There were no consistent short-term variations in fecal coliform or E. coli concentrations during the summer period or any significant relations between concentrations and stream discharge, based on these results.

Minnesota, Wisconsin↗

Sanitary quality of the Jordan River in Salt Lake County, Utah

This investigation of the sanitary quality of the Jordan River was conducted from July 1980 to October 1982 using indicator bacteria rather than specific pathogens. A serious sanitary problem was identified. Concentrations of total coliform bacteria often exceeded 5,000 colonies per 100 milliliters and concentrations of fecal coliform bacteria often exceeded 2,000 colonies per 100 milliliters in the lower reaches of the river. At times these levels were greatly exceeded. The most conspicuous aspect of the bacteriological data is its extreme variability. Seven waste-water treatment plants, seven major tributaries, numerous storm conduits, irrigation-return flow, and other sources all contribute to the dynamic system that determines the sanitary quality of the river. Because of this variability the sanitary quality of the river cannot be predicted at any one time. In general, concentrations of all three indicator bacteria increased in a downstream direction. Storm runoff from urban areas contributed large concentrations of indicator bacteria to the river. Regression analysis of 9 years of data collected at 1700 South Street showed a significant positive correlation between both fecal coliform and fecal streptococcal concentrations versus time. Concentrations of fecal coliform and fecal streptococci have both been increasing since 1974 at 1700 South Street. (USGS)

Water-Resources Investigations Report↗

Water resources of Oley Township, Berks County, Pennsylvania

Oley Township covers an area of 24 square miles, about half of which is underlain by highly permeable carbonate rocks. Nondomestic wells in these rocks typically have yields of 200 gallons per minute, and some wells yield more than 1,000 gallons per minute. Ground-water yield for Oley Township is about 0.5 million gallons per day per square mile. Thus, about 12 million gallons per day could be pumped from wells on a sustained basis. However, pumping this amount would adversely affect streamflow. A series of discharge measurements on Manatawny Creek in January 1983 showed that the creek was gaining approximately 12 cubic feet per second where it crosses the more- permeable carbonate rocks. Thus, the streams are directly connected to these aquifers. The northern and western parts of the township are mostly underlain by shale, quartzite, granite, gneiss, and carbonate rocks of low permeability, and some wells do not yield enough water for domestic supplies. A water-table map shows that two active quarries in low-permeability rocks have had little effect on the hydrologic system. Specific yields are about 4.5 percent for the carbonate rocks; 5 percent for quartzite, granite, and gneiss; 1 percent for the noncarbonate sedimentary rocks; and 1.5 percent for the Jacksonburg Limestone, which consists of argillaceous limestone. In 1982--a year of average precipitation--the ground-water contribution to total streamflow ranged from 56 to 88 percent. Basins with the highest percentage of carbonate rock contribute the largest amount of ground water to streamflow. Evapotranspiration averaged about 26 inches in 1982. Water loss was 32 inches in the Limekiln Creek basin; this suggests that about 6 inches of precipitation bypassed the Limekiln Creek gaging station as ground-water underflow. The most serious water-quality problems are excessive nitrate concentrations and bacterial contamination. Water from 3 of 19 wells in carbonate rocks had nitrate concentrations in excess of the U.S. Environmental Protection Agency maximum contaminant level of 10 milligrams per liter. Water from 5 of the 19 wells had fecal streptococci counts of more than 20 colonies per 100 milliliters. Although most agencies concerned with the protection of public health have not set limits for fecal streptococci, they are pathogenic, and their presence in drinking water is undesirable.

Water-Resources Investigations Report↗

Ground-Water Quality Reconnaissance, Tutuila, American Samoa, 1989

In May and July 1989, 19 ground-water samples were collected from 17 of the 35 water-supply wells on the island of Tutuila, American Samoa. Samples were analyzed for temperature, pH, hardness, specific conductance, turbidity, total and fecal coliform bacteria, major ions, major nutrients, 12 common metals, and 66 organic compounds. Chemical analysis of the water samples shows that, after chlorination, sampled ground water was in compliance with U.S. Environmental Protection Agency primary drinking-water regulations. Fourteen of the 19 samples were collected before the water was chlorinated in the water distribution system, and five were collected after chlorination. Seven of the 14 unchlorinated samples contained coliform bacteria and five of the seven contained fecal coliform bacteria, indicating potential contamination from disease-causing pathogens. All five of the chlorinated samples contained trace levels of trihalomethanes, but at concentrations below U.S. Environmental Protection Agency maximum contaminant levels. Trihalomethanes were the only organic constituents detected. Concentrations of total dissolved solids in the 19 samples ranged from about 100 to 2,400 milligrams per liter. Four samples had concentrations of total dissolved solids and chloride that exceeded the U.S. Environmental Protection Agency secondary drinking-water regulations of 500 and 250 milligrams per liter, respectively. For samples with less than 300 milligrams per liter of dissolved solids, sodium, calcium, magnesium, and bicarbonate were the dominant ions, indicating that they were derived from dissolution of minerals and carbon-dioxide enrichment of recharge water. For samples with greater than 300 milligrams per liter of dissolved solids, sodium and chloride were the dominant ions, indicating increased mixing of ground water with seawater.

Water-Resources Investigations Report↗

Effect of the Cedar River on the quality of the ground-water supply for Cedar Rapids, Iowa

The Surface Water Treatment Rule under the 1986 Amendment to the Safe Drinking Water Act requires that public-water supplies be evaluated for susceptibility to surface-water effects. The alluvial aquifer adjacent to the Cedar River is evaluated for biogenic material and monitored for selected water-quality properties and constituents to determine the effect of surface water on the water supply for the City of Cedar Rapids, Iowa. Results from monitoring of selected water-quality properties and constituents showed an inverse relation to river stage or discharge. Water-quality properties and constituents of the alluvial aquifer changed as water flowed from the river to the municipal well as a result of drawdown. The values of specific conductance, pH, temperature, and dissolved oxygen at observation well CRM-4 and municipal well Seminole 10 generally follow the trends of values for the Cedar River. Values at observation well CRM-3 and the municipal water-treatment plant showed very little correlation with values from the river. The traveltime of water through the aquifer could be an indication of the susceptibility of the alluvial aquifer to surface-water effects. Estimated traveltimes from the Cedar River to municipal well Seminole 10 ranged from 7 to 17 days. Above-normal streamflow and precipitation during the study could have increased the effect the river had on the alluvial aquifer and on the possibility of contamination by a pathogen. Microscopic particulate analysis of 29 samples found no Giardia cysts or Crytosporidium oocysts in water collected from municipal wells. Data also indicate that the aquifer is filtering out large numbers of algae, diatoms, rotifers, and nematodes as well as filtering out Cryptosporidium , Giardia , and other protozoa. The number of algae, diatoms, rotifers, protozoa, and vegetative debris for selected municipal wells tested showed at least a reduction to 1 per 1,000 of the number found in the river. A relative risk factor and a log-reduction rate were determined for the aquifer in the vicinity of selected wells. One municipal well had a high-risk factor, three other wells had a moderate-risk factor, and four wells had a low-risk factor. The filtering efficiency of the aquifer is equivalent to a 3 log-reduction rate or 99.99-percent reduction in particulates.

Iowa↗

Effects of hydrologic, biological, and environmental processes on sources and concentrations of fecal bacteria in the Cuyahoga River, with implications for management of recreational waters in Summit and Cuyahoga Counties, Ohio

Discharges of fecal bacteria (fecal coliform bacteria and Escherichia coli ) to the middle main stem of the Cuyahoga River from storm water, combined sewers, and incompletely disinfected wastewater have resulted in frequent exceedances of bacteriological water-quality standards in a 23-mile reach of the river that flows through the Cuyahoga Valley National Recreation Area. Contamination of the middle main stem of the Cuyahoga River by bacteria of fecal origin and subsequent transport to downstream areas where water-contact recreation is an important use of the river are a concern because of the potential public-health risk from the presence of enteric pathogens. Independent field investigations of bacterial decay, dilution, dispersion, transport, and sources, and bacterial contamination of streambed sediments, were completed in 1991-93 during periods of rainfall and runoff. The highest concentration of fecal coliform bacteria observed in the middle main stem during three transport studies exceeded the single-sample fecal coliform standard applicable to primary-contact recreation by a factor of approximately 1,300 and exceeded the Escherichia coli standard by a factor of approximately 8,000. The geometric-mean concentrations of fecal bacteria in the middle main stem were 6.7 to 12.3 times higher than geometric-mean concentrations in the monitored tributaries, and 1.8 to 7.0 times larger than the geometric-mean concentrations discharged from the Akron Water Pollution Control Station. Decay rates of fecal bacteria measured in field studies in 1992 ranged from 0.0018 per hour to 0.0372 per hour for fecal coliform bacteria and from 0.0022 per hour to 0.0407 per hour for Escherichia coli. Most of the decay rates measured in June and August were significantly higher than decay rates measured in April and October. Results of field studies demonstrated that concentrations of fecal coliform bacteria were 1.2 to 58 times higher in streambed sediments than in the overlying water. Sediments are likely to be a relatively less important source of fecal bacteria during rainfall and runoff in the middle main stem relative to bacterial loading from point sources. Numerical streamflow and transport simulation models were calibrated and verified with data collected during field studies. Of the constituents modeled, bacteria exhibited the poorest correspondence between observed and simulated values. The simulation results for a dye tracer indicated that the model reasonably reproduced the timing of dissolved constituents as well as dilution and dispersion effects. Calibrated and verified models for 1991 and 1992 data sets were used to simulate the improvements to bacteriological water quality that might result from reductions in concentrations of fecal bacteria discharged from two major sources. The model simulation resulting in the greatest improvement in bacteriological water-quality was one in which concentrations of fecal coliform bacteria and Escherichia coli were reduced by 90 percent in the Cuyahoga River at the Old Portage gaging station, and to geometric-mean bathing-water standards in the effluent of the Akron Water Pollution Control Station (BWS/90 scenario). Compared to the results of the base-simulation, when the BWS/90 scenario was applied in the 1991 model simulation, Escherichia coli concentrations were reduced 98.5 percent at Botzum, 97.5 percent at Jaite, and 91.1 percent at Independence. For 1992 model simulations, similar percent reductions in the concentrations of Escherichia coli were predicted at the three stream sites when the same reductions were applied to sources. None of the model simulations resulted in attainment of bacteriological water-quality standards. The potential benefits of source reductions to human health and recreational uses were estimated by comparing the number of illnesses per 1,000 people from concentrations of Escherichia coli associated with the BWS/90 simulation, with the base simulation, and with the geometric-mean standard for Escherichia coli. The predicted 22 to 26 illnesses per 1,000 people predicted by the E. coli concentrations resulting from BWS/90 simulation are 2.8 to 3.3 times higher than the 8 illnesses per 1,000 people associated with the geometric-mean primary-contact water-quality standard for Escherichia coli. Risks associated with the base simulation are 4.6 to 4.9 times higher than that associated with the geometric-mean primary- contact water-quality standard for Escherichia coli. The illness risks predicted from the BWS/90 scenario, although larger than acceptable, would nevertheless be an improvement over conditions that were encountered during field studies in 1991-93.

Ohio↗

Environmental setting and water-quality issues in the lower Tennessee River basin

The goals of the National Water-Quality Assessment Program are to describe current water-quality conditions for a large part of the Nation's water resources, identify water-quality changes over time, and identify the primary natural and human factors that affect water quality. The lower Tennessee River Basin is one of 59 river basins selected for study. The water-quality assessment of the lower Tennessee River Basin study unit began in 1997. The lower Tennessee River Basin study unit encompasses an area of about 19,500 square miles and extends from Chattanooga, Tennessee, to Paducah, Kentucky. The study unit had a population of about 1.5 million people in 1995. The study unit was subdivided into subunits with relatively homogeneous geology and physiography. Subdivision of the study unit creates a framework to assess the effects of natural and cultural settings on water quality. Nine subunits were delineated in the study unit; their boundaries generally coincide with level III and level IV ecoregion boundaries. The nine subunits are the Coastal Plain, Transition, Western Highland Rim, Outer Nashville Basin, Inner Nashville Basin, Eastern Highland Rim, Plateau Escarpment and Valleys, Cumberland Plateau, and Valley and Ridge.The lower Tennessee River Basin consists of predominantly forest (51 percent) and agricultural land (40 percent). Activities related to agricultural land use, therefore, are the primary cultural factors likely to have a widespread effect on surface- and ground-water quality in the study unit. Inputs of total nitrogen and phosphorus from agricultural activities in 1992 were about 161,000 and 37,900 tons, respectively. About 3.7 million pounds (active ingredient) of pesticides was applied to crops in the lower Tennessee River Basin in 1992. State water-quality agencies identified nutrient enrichment and pathogens as water-quality issues affecting both surface and ground water in the lower Tennessee River Basin. Water-quality data collected by State and Federal agencies between 1980 and 1996 were summarized to characterize surface- and ground-water quality of the subunits with respect to these issues. Median concentrations of nitrogen species generally were less than 1 milligram per liter in surface and ground water in all subunits, and were highest throughout the subunits that had the largest percentages of agricultural land use. Median phosphorus concentrations also were less than 1 milligram per liter in all subunits. Phosphatic limestones present in two subunits had a larger effect on phosphorus concentrations in surface and ground water than did the amount of agricultural land use in these subunits. Median counts of fecal coliform were higher in surface water than in ground water in all subunits. The highest median counts in surface water were in the Valley and Ridge (7,500 colonies per 100 milliliters) and the Outer Nashville Basin subunits (5,000 colonies per 100 milliliters). Highest median counts in ground water were in the Inner and Outer Nashville Basin subunit. Natural setting likely has an important effect with respect to fecal contamination of surface and ground water in the lower Tennessee River Basin.

Alabama, Georgia, Kentucky, Mississippi, Tennessee↗

Quantity and quality of seepage from two earthen basins used to store livestock waste in southern Minnesota during the first year of operation, 1997-98

Numerous earthen basins have been constructed in Minnesota for storage of livestock waste. Typically, these basins are excavated pits with partially above-grade, earth-walled embankments and compacted clay liners. Some have drain tile installed around them to prevent shallow ground and soil water to discharge into the basins. Environmental concerns associated with the waste include contamination of ground water by nitrogen compounds and pathogens. The U.S. Geological Survey, in cooperation with the MFC A (Minnesota Pollution Control Agency), studied the quantity and quality of seepage from two earthen basins used to store livestock waste in southern Minnesota during the first year of operation. One basin (site A), located at a small dairy farm, holds a manuresilage mixture, milkhouse wastewater, and local runoff; the other basin (site B), located at a large hog farm, holds a manure-water mixture from a nearby gestation barn. Monitoring systems were installed below compacted clay liners in portions of the sidewalls and bottoms of the basins to determine the quantity and quality of the seepage. Total seepage flow from the site A basin ranged from about 900 to 2,400 gal/d (gallons per day) except during April 1998 when the flow increased to about 4,200 gal/d. Seepage flow in areal units, which closely correlated with flow in gal/d, generally ranged from about 0.07 to 0.18 in./d (inches per day), which exceeded the recommended maximum design rate of 0.018 in./d established by the MPCA. Seepage flow commonly was greater through the sidewalls than through the bottom. Seepage from the site A basin (based on 11 samples each from the bottom and sidewall) had chloride concentrations of 220-350 mg/L (milligrams per liter); ammonium-N (nitrogen) concentrations of 2.40 mg/L or less (except for one concentration of 18.4 mg/L); nitrate-N concentrations of 5.24 mg/L or less; and organic-N concentrations of 6.97 mg/L or less. Ground water would be enriched in chloride and diluted in inorganic-N from mixing with basin seepage. Fecal Coliform bacteria, although abundant in the basin wastewater, were present in very small amounts in the seepage. Total seepage flow from the site B basin generally ranged from 400 to 2,200 gal/d except during 1-month and 3-month periods when the flow ranged from about 3,800 to 6,200 gal/d. Seepage flow in areal units generally ranged from about 0.025 to 0.15 in./d, and, as at the site A basin, exceeded the MPCA recommended maximum design rate of 0.018 in./d. Seepage flow in areal units generally correlated with the flow in gal/d except through the sidewalls when the basin was unfilled. Except during the first three months of the study, seepage flow was greater through the sidewalls than through the bottom. Seepage from the site B basin (based on 10 samples each from the bottom and sidewall) had chloride concentrations of 11 to 100 mg/L; ammonium-N concentrations of 2.58 mg/L or less; nitrate-N concentrations of 25.7 mg/L or less (except for one concentration of 146 mg/L); and organic-N concentrations of 0.92 mg/L or less. Nitrate-N concentrations in the seepage exceeded the U.S. Environmental Protection Agency (1996). MCL (maximum contaminant level) of 10 mg/L in 17 of 22 samples. Background ground-water quality, however, indicated that nitrate-N concentrations were greater than the MCL prior to operation of the basin. Fecal Coliform bacteria, as at the site A basin, were abundant in the basin wastewater, but not in the seepage.

Minnesota↗

The status of mussel health assessment and a path forward

Declines of freshwater mussel (order Unionida) populations worldwide are attributed to habitat degradation, pollution, and invasive species, among other factors. However, these purported causes do not fully explain the enigmatic decline and large-scale die-offs of mussels that have occurred in assumedly “healthy” streams across a wide geographic region. The roles of the microbiota and pathogens in mussel health has been understudied, and, as a result, few references exist to compare the microbiota of healthy mussels to that of stressed or dying mussels. Captive propagation and stocking programs have expanded across the globe without standard diagnostic protocols to assess health or potential diseases in hatchery-reared or wild stocks. Introduction of nonindigenous species, contaminants of emerging concern and anthropogenic climate change could adversely alter underlying variables that support mussel health such as nutrient and microbial composition, in addition to increased risk for outbreaks of opportunistic disease and emergent diseases in freshwater mussel populations. We suggest a coordinated, collaborative, and multidisciplinary effort to advance methods for assessing freshwater mussel health. We identify research and resources needed to answer central questions surrounding mussel health including identifying potential agents of disease, defining clinical signs of declining condition, refining stress-specific biomarkers for health assessment, and developing protocols specific for mussels.

Freshwater Mollusk Biology and Conservation↗

Bat flight and zoonotic viruses

Bats are sources of high viral diversity and high-profile zoonotic viruses worldwide. Although apparently not pathogenic in their reservoir hosts, some viruses from bats severely affect other mammals, including humans. Examples include severe acute respiratory syndrome coronaviruses, Ebola and Marburg viruses, and Nipah and Hendra viruses. Factors underlying high viral diversity in bats are the subject of speculation. We hypothesize that flight, a factor common to all bats but to no other mammals, provides an intensive selective force for coexistence with viral parasites through a daily cycle that elevates metabolism and body temperature analogous to the febrile response in other mammals. On an evolutionary scale, this host–virus interaction might have resulted in the large diversity of zoonotic viruses in bats, possibly through bat viruses adapting to be more tolerant of the fever response and less virulent to their natural hosts.

Emerging Infectious Diseases↗

Paranannizziopsis spp. Infection in Wild Vipers, Europe

We describe the first detection of Paranannizziopsis sp. in a wild population of vipers in Europe. Fungal infections were severe and one individual likely died from infection. Surveillance efforts are needed to better understand the threat of this pathogen to snake conservation.

Emerging Infectious Diseases↗

Microparasite ecology and health status of common bluestriped snapper Lutjanus kasmira from the Pacific Islands

Common bluestriped snappers Lutjanus kasmira were intentionally introduced into Hawaii from the South Pacific in the 1950s and have become well established throughout the archipelago. We examined health, prevalence and infection intensity of 2 microparasites, coccidia and epitheliocystis-like organisms (ELO), in L. kasmira from their introduced and native range including the islands where translocated fish originated (Tahiti and Marquesas Islands, French Polynesia) and from several other islands (American Samoa, Fiji and New Caledonia). In addition, we did a longitudinal survey of these parasites in the introduced range. Coccidia and ELO were widely distributed and were found on all islands except for New Caledonia. Health indices, as measured by overall organ lesions, body condition and parasite intensity, indicated that fish from Samoa were the least healthy, and fish from Midway (Hawaiian Archipelago) were the healthiest. Microparasite diversity was highest on Midway and Hawaii and lowest on New Caledonia. Infection of coccidia followed an asymptotic size–prevalence curve, whereas that of ELO peaked at larger size classes (27.8 cm). Prevalence and aggregation of both parasites in the host varied dynamically over 8 yr, with prevalence and aggregation of coccidia being consistently higher and lower, respectively, than ELO. We hypothesize that these parasites are enzootic to the Hawaiian Islands and were not introduced with fish from Tahiti or the Marquesas Islands. Host response and aggregation parameters suggest that coccidia exert a negative effect on their host and probably have an indirect life cycle, whereas ELO appears less pathogenic and has a direct life cycle.

Aquatic Biology↗

Quantitative PCR detection of Batrachochytrium dendrobatidis DNA from sediments and water

The fungal pathogen Batrachochytrium dendrobatidis (Bd) causes chytridiomycosis, a disease implicated in amphibian declines on 5 continents. Polymerase chain reaction (PCR) primer sets exist with which amphibians can be tested for this disease, and advances in sampling techniques allow non-invasive testing of animals. We developed filtering and PCR based quantitative methods by modifying existing PCR assays to detect Bd DNA in water and sediments, without the need for testing amphibians; we tested the methods at 4 field sites. The SYBR based assay using Boyle primers (SYBR/Boyle assay) and the Taqman based assay using Wood primers performed similarly with samples generated in the laboratory (Bd spiked filters), but the SYBR/Boyle assay detected Bd DNA in more field samples. We detected Bd DNA in water from 3 of 4 sites tested, including one pond historically negative for chytridiomycosis. Zoospore equivalents in sampled water ranged from 19 to 454 l-1 (nominal detection limit is 10 DNA copies, or about 0.06 zoospore). We did not detect DNA of Bd from sediments collected at any sites. Our filtering and amplification methods provide a new tool to investigate critical aspects of Bd in the environment.

Diseases of Aquatic Organisms↗

Chronic and persistent viral hemorrhagic septicemia virus infections in Pacific herring

Chronic viral hemorrhagic septicemia virus (VHSV) infections were established in a laboratory stock of Pacific herring Clupea pallasii held in a large-volume tank supplied with pathogen-free seawater at temperatures ranging from 6.8 to 11.6°C. The infections were characterized by viral persistence for extended periods and near-background levels of host mortality. Infectious virus was recovered from mortalities occurring up to 167 d post-exposure and was detected in normal-appearing herring for as long as 224 d following initial challenge. Geometric mean viral titers were generally as high as or higher in brain tissues than in pools of kidney and spleen tissues, with overall prevalence of infection being higher in the brain. Upon re-exposure to VHSV in a standard laboratory challenge, negligible mortality occurred among groups of herring that were either chronically infected or fully recovered, indicating that survival from chronic manifestations conferred protection against future disease. However, some survivors of chronic VHS infections were capable of replicating virus upon re-exposure. Demonstration of a chronic manifestation of VHSV infection among Pacific herring maintained at ambient seawater temperatures provides insights into the mechanisms by which the virus is maintained among populations of endemic hosts.

Diseases of Aquatic Organisms↗

Larval and juvenile Pacific herring Clupea pallasii are not susceptible to infectious hematopoietic necrosis under laboratory conditions

Infectious hematopoietic necrosis (IHN) leads to periodic epidemics among certain wild and farmed fish species of the Northeast (NE) Pacific. The source of the IHN virus (IHNV) that initiates these outbreaks remains unknown; however, a leading hypothesis involves viral persistence in marine host species such as Pacific herring Clupea pallasii. Under laboratory conditions we exposed specific pathogen-free (SPF) larval and juvenile Pacific herring to 10 3 to 10 4 plaque-forming units (pfu) of IHNV ml –1 by waterborne immersion. Cumulative mortalities among exposed groups were not significantly different from those of negative control groups. After waterborne exposure, IHNV was transiently recovered from the tissues of larvae but absent in tissues of juveniles. Additionally, no evidence of viral shedding was detected in the tank water containing exposed juveniles. After intraperitoneal (IP) injection of IHNV in juvenile herring with 10 3 pfu, IHNV was recovered from the tissues of sub-sampled individuals for only the first 5 d post-exposure. The lack of susceptibility to overt disease and transient levels of IHNV in the tissues of exposed fish indicate that Pacific herring do not likely serve a major epizootiological role in perpetuation of IHNV among free-ranging sockeye salmon Oncorhynchus nerka and farmed Atlantic salmon Salmo salar in the NE Pacific.

Diseases of Aquatic Organisms↗

Universal reverse-transcriptase real-time PCR for infectious hematopoietic necrosis virus (IHNV)

Infectious hematopoietic necrosis virus (IHNV) is an acute pathogen of salmonid fishes in North America, Europe and Asia and is reportable to the World Organization for Animal Health (OIE). Phylogenetic analysis has identified 5 major virus genogroups of IHNV worldwide, designated U, M, L, E and J; multiple subtypes also exist within those genogroups. Here, we report the development and validation of a universal IHNV reverse-transcriptase real-time PCR (RT-rPCR) assay targeting the IHNV nucleocapsid (N) gene. Properties of diagnostic sensitivity (DSe) and specificity (DSp) were defined using laboratory-challenged steelhead trout Oncorhynchus mykiss, and the new assay was compared to the OIE-accepted conventional PCR test and virus isolation in cell culture. The IHNV N gene RT-rPCR had 100% DSp and DSe and a higher estimated diagnostic odds ratio (DOR) than virus culture or conventional PCR. The RT-rPCR assay was highly repeatable within a laboratory and highly reproducible between laboratories. Field testing of the assay was conducted on a random sample of juvenile steelhead collected from a hatchery raceway experiencing an IHN epizootic. The RT-rPCR detected a greater number of positive samples than cell culture and there was 40% agreement between the 2 tests. Overall, the RT-rPCR assay was highly sensitive, specific, repeatable and reproducible and is suitable for use in a diagnostic setting.

Diseases of Aquatic Organisms↗

Testing of candidate non-lethal sampling methods for detection of Renibacterium salmoninarum in juvenile Chinook salmon Oncorhynchus tshawytscha

Non-lethal pathogen testing can be a useful tool for fish disease research and management. Our research objectives were to determine if (1) fin clips, gill snips, surface mucus scrapings, blood draws, or kidney biopsies could be obtained non-lethally from 3 to 15 g Chinook salmon Oncorhynchus tshawytscha , (2) non-lethal samples could accurately discriminate between fish exposed to the bacterial kidney disease agent Renibacterium salmoninarum and non-exposed fish, and (3) non-lethal samples could serve as proxies for lethal kidney samples to assess infection intensity. Blood draws and kidney biopsies caused ≥5% post-sampling mortality (Objective 1) and may be appropriate only for larger fish, but the other sample types were non-lethal. Sampling was performed over 21 wk following R. salmoninarum immersion challenge of fish from 2 stocks (Objectives 2 and 3), and nested PCR (nPCR) and real-time quantitative PCR (qPCR) results from candidate non-lethal samples were compared with kidney tissue analysis by nPCR, qPCR, bacteriological culture, enzyme-linked immunosorbent assay (ELISA), fluorescent antibody test (FAT) and histopathology/immunohistochemistry. R. salmoninarum was detected by PCR in >50% of fin, gill, and mucus samples from challenged fish. Mucus qPCR was the only non-lethal assay exhibiting both diagnostic sensitivity and specificity estimates >90% for distinguishing between R. salmoninarum -exposed and non-exposed fish and was the best candidate for use as an alternative to lethal kidney sample testing. Mucus qPCR R. salmoninarum quantity estimates reflected changes in kidney bacterial load estimates, as evidenced by significant positive correlations with kidney R. salmoninarum infection intensity scores at all sample times and in both fish stocks, and were not significantly impacted by environmental R. salmoninarum concentrations.

Diseases of Aquatic Organisms↗