A herpesvirus was isolated from adult koi, a strain of common carp Cyprinus carpio, suffering mass mortality in two outbreaks-one in the mid-Atlantic region of the United States and the second in Israel. The principal external signs of dying fish were pale and irregularly colored gills. There were few consistent internal signs in either outbreak. The most prominent microscopic lesions were in the gills, where hyperplasia and necrosis of the epithelium were severe. Other lesions included interstitial nephritis, splenitis, and enteritis. Affected cells often contained nuclei with marginated chromatin and faint intranuclear inclusions. Typical herpesvirus particles were present in branchial epithelial cells, hepatocytes, and among circulating leukocytes. Inoculations of the koi fin (KF-1) cell line with tissue extracts from the gill and kidney-spleen resulted in cytopathic effects characterized by severe vacuolation first detected after 7 d incubation at 20°C. Exposures of adult koi to the herpesvirus as propagated in KF-1 cells by bath or intraperitoneal injections resulted in 80-100% mortality during a 26-d period, and the virus was reisolated from the gill, kidney, liver, spleen, intestine, and brain of dead fish. The viral agents from koi in Israel and the United States appear to be similar if not identical; both could be distinguished from Herpesvirus cyprini by indirect fluorescent antibody tests with rabbit anti-H. cyprini serum. Other factors should be examined but we strongly suspect that this newly recognized koi herpesvirus (KHV) has the potential to be a significant cause of mortality among koi and presumably common carp.

Heart and skeletal muscle inflammation (HSMI) is a significant and often fatal disease of cultured Atlantic salmon in Norway. The consistent presence of Piscine orthoreovirus (PRV) in HSMI diseased fish along with the correlation of viral load and antigen with development of lesions has supported the supposition that PRV is the etiologic agent of this condition; yet the absence of an in vitro culture system to demonstrate disease causation and the widespread prevalence of this virus in the absence of disease continues to obfuscate the etiological role of PRV with regard to HSMI. In this study, we explore the infectivity and disease causing potential of PRV from western North America—a region now considered endemic for PRV but without manifestation of HSMI—in challenge experiments modeled upon previous reports associating PRV with HSMI. We identified that western North American PRV is highly infective by intraperitoneal injection in Atlantic salmon as well as through cohabitation of both Atlantic and Sockeye salmon. High prevalence of viral RNA in peripheral blood of infected fish persisted for as long as 59 weeks post-challenge. Nevertheless, no microscopic lesions, disease, or mortality could be attributed to the presence of PRV, and only a minor transcriptional induction of the antiviral Mx gene occurred in blood and kidney samples during log-linear replication of viral RNA. Comparative analysis of the S1 segment of PRV identified high similarity between this North American sequence and previous sequences associated with HSMI, suggesting that factors such as viral co-infection, alternate PRV strains, host condition, or specific environmental circumstances may be required to cause this disease.

Development of pink salmon (Oncorhynchus gorbuscha) incubating in gravel contaminated with weathered Prudhoe Bay crude oil was retarded at concentrations as low as 55.2 μg oil/g gravel. Larvae exposed to various levels of oil contamination were sampled 4 weeks before emergence, at emergence, and 13 days after emergence for histopathology (quantitative and semiquantitative) and cytochrome P4501A (CYP1A) induction (using immunohistochemical staining). A subset of postemergent fish was not fed. Hydrocarbon analysis by gas chromatography and mass spectroscopy revealed that tissue uptake of polynuclear aromatic hydrocarbons (PAH) was mediated by oil's dissolution in water, with significant biological effects when the peak total PAH concentration in water was as low as 4.4 μg/L. Oil-related effects included induction of CYP1A, development of ascites, and increased mortality. Several oil-related changes were indicative of premature emergence. Compared with control fish, for example, exposed fish of the same age and emerging on the same day had greater amounts of yolk and hepatocellular glycogen, increased apoptosis of gonadal cells and midventral skin cells, and less food in the gastrointestinal tract. Histological features were similar within groups of larvae sampled 4 weeks before and 13 days after emergence, and oil-induced changes were not affected by feeding during the first 13 days after emergence. Increased gonadal cell apoptosis may be related to later reproductive impairment documented in field studies of pink salmon up to 4 years after the Exxon Valdez oil spill.

We have carried out the first major infectivity trial to examine differential genetic resistance in fish for pathogens. We used captive-bred, endangered winter-run chinook salmon (Oncorhynchus tshawytscha) to determine resistance to three pathogens: the bacterium, Listonella (Vibrio) anguillarum, infectious hematopoietic necrosis virus (IHNV), and Myxobolus cerebralis, the parasite that causes whirling disease. We compared resistance to these three pathogens between inbred and outbred salmon and between siblings that were heterozygous or homozygous for a class II gene in the major histocompatibility complex (MHC). In two of five different comparisons, we found significant genetic effects on disease resistance. First, MHC heterozygotes had a higher survival than MHC homozygotes when exposed to IHNV and the selection disadvantage of homozygotes was estimated to be 8.5%. Second, outbred fish had a higher resistance (or lower infection severity) than inbred fish when exposed to M. cerebralis. Using a quantitative genetics approach, it appears that there are slightly more than three gene equivalents segregating that would result in no resistance to M. cerebralis when homozygous. Overall, our investigation suggests that pathogen susceptibility in the winter-run chinook salmon will increase if further genetic variation is lost in this endangered species.

Increased farm salmon production has heightened concerns about the association between disease on farm and wild fish. The controversy is particularly evident in the Broughton Archipelago of Western Canada, where a high prevalence of sea lice (ectoparasitic copepods) was first reported on juvenile wild pink salmon (Oncorhynchus gorbuscha) in 2001. Exposure to sea lice from farmed Atlantic salmon (Salmo salar) was thought to be the cause of the 97% population decline before these fish returned to spawn in 2002, although no diagnostic investigation was done to rule out other causes of mortality. To address the concern that sea lice from fish farms would cause population extinction of wild salmon, we analyzed 10-20 y of fish farm data and 60 y of pink salmon data. We show that the number of pink salmon returning to spawn in the fall predicts the number of female sea lice on farm fish the next spring, which, in turn, accounts for 98% of the annual variability in the prevalence of sea lice on outmigrating wild juvenile salmon. However, productivity of wild salmon is not negatively associated with either farm lice numbers or farm fish production, and all published field and laboratory data support the conclusion that something other than sea lice caused the population decline in 2002. We conclude that separating farm salmon from wild salmon-proposed through coordinated fallowing or closed containment-will not increase wild salmon productivity and that medical analysis can improve our understanding of complex issues related to aquaculture sustainability.B ecause salmon aquaculture production has rapidly increased over the past three decades, the potential for environmental impacts of salmon farms has generated heightened scientific and public interest (1, 2). One concern about salmon farms is that they are the source of ectoparasitic sea lice infestations that might reduce the marine survival of wild salmon (3, 4). In the Broughton Archipelago region of Western Canada (Fig. S1), farming of Atlantic salmon (Salmo salar) began in the late 1980s, and annual farm salmon production increased steadily to 17 Gg by 1999 (Fig. S2). Pink salmon (Oncorhynchus gorbuscha) is the most abundant wild salmon species in the Broughton Archipelago; they enter the marine environment at a very small size (0.2 g), and they return to natal streams to spawn 2 y after their parents (5). Because age at maturity never varies, they have distinct evenand odd-year populations ( Fig. S2 and SI Text). Record high numbers of pink salmon returned to spawn in rivers of the Broughton Archipelago in 2000 and 2001 (Dataset S1), but these returns were followed by population decline of 97% in 2002 and 88% in 2003 (Figs. S2 and S3 and SI Text). When juvenile pink salmon in the Broughton Archipelago were first examined for sea lice in June 2001, more than 90% were infested-leading to the hypothesis that sea lice from fish farms were the cause of population collapse in 2002 (4).Adult pink salmon are a natural host for the sea louse species Lepeophtheirus salmoni...