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Immunostimulants, adjuvants, and vaccine carriers in fish: Applications to aquaculture

Use of immunostimulants, adjuvants, and vaccine carriers in fish culture offers a wide range of attractive methods for inducing and building up protection against diseases. Immunostimulants and adjuvants can be administered before, with, or after vaccines to amplify the specific immune response generating elevations of circulating antibody titers and numbers of plaque-forming cells. Special applications of immunostimulants include assisting shower or other regimens to increase topical uptake of vaccines. In addition, immunostimulants may be used alone, inducing elevated activities in the nonspecific defense mechanisms such as increased oxidative activity of neutrophils, augmented engulfment activity of phagocytic cells, or potentiating cytotoxic cells. In cases where disease outbreaks are cyclical and can be predicted, losses may be reduced by elevating the nonspecific defense mechanisms, and the immunostimulants may be used in anticipation of events to prevent losses from diseases. Complete Freund's adjuvant was one of the first immunostimulants used in animals to elevate the specific immune response, and it has also been successfully used in conjunction with injection of fish bacterins. Other adjuvants, immunostimulants, and biological response modifiers that have been used in fisheries research include levamisole, salt baths, and bacterial lipopolysaccharides. Vaccines have been adsorbed to inert particles, such as bentonite on latex beads, to carry the immunogens to maximize in vivo uptake for bath immunization and to facilitate in vitro phagocytosis. Each substance presents special problems in timing and method of administration (injection, immersion, oral—by feed—or flush treatments), dosage adjustments for size and fish species, storage stability, and cost. An additional consideration is that the nonspecific defense mechanisms and immune responses in fish are highly variable among individuals and statistical validation requires appropriate sample numbers and carefully controlled experiments. This article reviews the literature and present concepts of use of immunostimulants, adjuvants, and vaccine carriers in fish. Cautions for use are noted, as some of these potent substances can suppress or alter biological pathways if used inappropriately. Recent research, defining pathways of the action of immunostimulants, adjuvants, and vaccine carriers, helps explain how these substances activate the protective mechanisms in fish. In addition, immunostimulants used alone hold tremendous potential for use in fish farms, hatcheries, and aquaculture facilities to reduce losses from infectious diseases. Research on the immunostimulant, levamisole, and the light oil adjuvants for use in food fish is in progress. Applications for use of these immunostimulants are proposed.

Annual Review of Fish Diseases↗

Recent advances in detection and control of infectious hematopoietic necrosis virus in aquaculture

Infectious hematopoietic necrosis (IHN) is one of the most important viral diseases of salmon and trout reared in culture. The disease remains untreatable with avoidance being the only control measure. Much has been learned about the chemical, physical, and serological characteristics of the rhabdovirus causing IHN, but critical gaps exist in our understanding of the biology of the virus in nature. The tools of molecular biology have provided improved methods for detection of pathogens and new strategies for control of viral diseases. This paper reviews several recent improvements in methods for detecting infectious hematopoietic necrosis virus including the application of enzyme-linked immunosorbent assays, development of monoclonal antibodies and DNA probes, and use of the polymerase chain reaction. New strategies for control of IHN through the use of better water treatment, more resistant fish, antiviral drugs or chemicals, and new generation vaccines are discussed.

Annual Review of Fish Diseases↗

The role of the diagnostic laboratory in fish disease control

The diagnostic laboratory is becoming increasingly important to the productivity and profitability of hatcheries, fish farms, and aquaculture stations. The basic function of the laboratory personnel is to isolate and identify viral, bacterial, protozoan, and other fish pathogens present in cultured and feral fish stocks. New, rapid and accurate methods for the detection and identification of fish disease agents based on immunological, biochemical, and physiological assays are becoming commonly used. Nearly every North American state or province and many foreign countries have fish health regulations that require inspection of stocks for certain disease agents before the fish are shipped into their areas. Decisions from the diagnostic laboratory on identification, treatment, guaranteed isolation, immunization, and disposal of fish populations affect administrative directives, hatchery placement, and national and international transportation of fish and fish products. This paper reviews concepts and describes the equipment, supplies, biologics, and media needed for the basic diagnostic laboratory. Information management, including training of staff, certification procedures, and quality control are also discussed.

Annual Review of Fish Diseases↗

Host defense mechanisms of Cephalopods

Humoral and cellular mechanisms of defense have been described for cephalopods, a relatively advanced group of mollusks. Typical of other mollusks, cephalopod agglutinins are the most documented component of humoral immunity. Lectins, which have agglutinating properties, have been described and characterized from octopuses. Agglutinins from cephalopod hemolymph have also been shown to agglutinate a variety of vertebrate red blood cells, as well as potential bacterial pathogens. Hemocytes are the primary component of cellular immunity. Although the hemocyte role in phagocytosis has been extensively studied in other mollusks, the mechanisms of phagocytosis have not been described extensively for cephalopods. Cephalopod hemocytes have phagocytic capabilities and may function in encapsulation and neutralization of foreign substances; however, the effects of environmental factors and the full extent of phagocytic capabilities of cephalopod hemocytes have not been reported. Hemocytes from cephalopods have a role in wound healing and inflammation which have been reported in detail by several investigators.

Annual Review of Fish Diseases↗

Viral hemorrhagic septicemia virus in North America

The first detections of viral hemorrhagic septicemia virus (VHSV) in North America were in Washington State from adult coho ( Oncorhynchus kisutch ) and chinook ( O. tshawytscha ) salmon in 1988. Subsequently, VHSV was isolated from adult coho salmon returning to hatcheries in the Pacific Northwest in 1989, 1991 and 1994. These isolates represented a strain of VHSV that was genetically different from European VHSV as determined by DNA sequence analysis and T1 ribonuclease fingerprinting. The North American strain of VHSV was also isolated from skin lesions of Pacific cod ( Gadus macrocephalus ) taken from Prince William Sound (PWS), Alaska in 1990, 1991 and 1993. In 1993 and 1994, the virus was isolated from Pacific herring ( Clupea harengus pallasi ) in Alaskan waters of PWS, Kodiak Island, Auke Bay and Port Frederick. During 1993 and 1994 the herring fishery in PWS failed from a probable complex of environmental stressors but VHSV isolates were associated with hemorrhages of the skin and fins in fish that returned to spawn. Also in 1993 and 1994, VHSV was isolated from apparently healthy stocks of herring in British Columbia, Canada and Puget Sound, Washington. Thus, the North American strain of VHSV is enzootic in the Northeastern Pacific Ocean among Pacific herring stocks with Pacific cod serving as a secondary reservoir. Although the North American strain of the virus appears to be moderately pathogenic for herring, causing occasional self-limiting epizootics, it was shown to be relatively avirulent for several species of salmonids. Pacific herring are common prey for cod and salmon and were most probably the source of the VHSV isolates from the adult salmon returning to spawn in rivers or at hatcheries in Washington State. Compelling circumstances involving the European isolates of VHSV suggest that this strain of the virus also is enzootic among marine fish in the Atlantic Oean. The highly pathogenic nature of the European strain of VHSV for salmonid fish may be the result of the exposure of rainbow trout ( O. mykiss ), an introduced species, in a stressful environment of intensive culture and the high rate of mutation inherent in all rhabdoviruses. Consequently, we recommend that efforts be made to eradicate the North American strain of VHSV when detected in live salmonids to reduce the possibility of its evolution into a more virulent salmonid virus.

Annual Review of Fish Diseases↗

Marine infectious disease ecology

To put marine disease impacts in context requires a broad perspective on the roles infectious agents have in the ocean. Parasites infect most marine vertebrate and invertebrate species, and parasites and predators can have comparable biomass density, suggesting they play comparable parts as consumers in marine food webs. Although some parasites might increase with disturbance, most probably decline as food webs unravel. There are several ways to adapt epidemiological theory to the marine environment. In particular, because the ocean represents a three-dimensional moving habitat for hosts and parasites, models should open up the spatial scales at which infective stages and host larvae travel. In addition to open recruitment and dimensionality, marine parasites are subject to fishing, filter feeders, dosedependent infection, environmental forcing, and death-based transmission. Adding such considerations to marine disease models will make it easier to predict which infectious diseases will increase or decrease in a changing ocean.

Annual Review of Ecology, Evolution, and Systemati↗

Are diseases increasing in the ocean?

Many factors (climate warming, pollution, harvesting, introduced species) can contribute to disease outbreaks in marine life. Concomitant increases in each of these makes it difficult to attribute recent changes in disease occurrence or severity to any one factor. For example, the increase in disease of Caribbean coral is postulated to be a result of climate change and introduction of terrestrial pathogens. Indirect evidence exists that ( a ) warming increased disease in turtles; ( b ) protection, pollution, and terrestrial pathogens increased mammal disease; ( c ) aquaculture increased disease in mollusks; and ( d ) release from overfished predators increased sea urchin disease. In contrast, fishing and pollution may have reduced disease in fishes. In other taxa (e.g., sea grasses, crustaceans, sharks), there is little evidence that disease has changed over time. The diversity of patterns suggests there are many ways that environmental change can interact with disease in the ocean.

Annual Review of Ecology, Evolution, and Systemati↗

Factors affecting post-challenge survival of Flavobacterium psychrophilum in susceptible rainbow trout from the literature

Infectious bacterial pathogens are a concern for aquaculture as estimates suggest that billions of US dollars are lost annually in aquaculture due to disease. One of the most prevalent salmonid pathogens is the bacterium Flavobacterium psychrophilum that causes bacterial coldwater disease. We reviewed the published F. psychrophilum literature and conducted a Bayesian analysis to examine large-scale patterns in rainbow trout ( Oncorhynchus mykiss ) mortality associated with laboratory challenge. We incorporated factors that were common across a majority of the laboratory exposure studies and these included bacterial dose, culture time, exposure method, bacterial isolate, experimental duration, and fish weight. The comparison showed that injection as the exposure method produced higher mortality than bath immersion, bacterial isolates differed in their effect on mortality, and bacterial dose has an interactive effect with fish weight and exposure method. Our comparison allows for inference on factors affecting rainbow trout mortality due to exposure to F. psychrophilum and suggests avenues to further optimize research protocols to better reach study goals.

Pathogens↗

Infectious diseases affect marine fisheries and aquaculture economics

Seafood is a growing part of the economy, but its economic value is diminished by marine diseases. Infectious diseases are common in the ocean, and here we tabulate 67 examples that can reduce commercial species' growth and survivorship or decrease seafood quality. These impacts seem most problematic in the stressful and crowded conditions of aquaculture, which increasingly dominates seafood production as wild fishery production plateaus. For instance, marine diseases of farmed oysters, shrimp, abalone, and various fishes, particularly Atlantic salmon, cost billions of dollars each year. In comparison, it is often difficult to accurately estimate disease impacts on wild populations, especially those of pelagic and subtidal species. Farmed species often receive infectious diseases from wild species and can, in turn, export infectious agents to wild species. However, the impact of disease export on wild fisheries is controversial because there are few quantitative data demonstrating that wild species near farms suffer more from infectious diseases than those in other areas. The movement of exotic infectious agents to new areas continues to be the greatest concern.

Annual Review of Marine Science↗

Introduction [to Issue 3]

Artificial propagation of aquatic organisms is increasing globally and currently accounts for approximately 32% of total world fishery production (Vannuccini, 2004). Between 1970 and 2000, aquaculture production of salmonids has grown from less than 200,000 metric tons per year to over 1.5 million metric tons (Tacon, 2003). In 1995, the number of Atlantic salmon (Salmo salar) far exceeded the carrying capacity of salmon in the wild but over 94% of all adult Atlantic salmon were in aquaculture environments (Gross, 1998). Since the 1970’s, concerns have arisen about interactions of hatchery and wild produced salmonids in native habitats. In response, research has addressed concerns about domestication and genetic impacts (Reisenbichler and McIntyre, 1977; Hindar et al., 1991; Waples, 1991; Clifford et al., 1998a; Fleming et al., 2000, 2002; Hard et al., 2000), transfer of disease and parasites (Johnsen and Jensen, 1994; Bakke and Harris, 1998), behavior of spawning adults (Fleming and Gross, 1992; Fleming et al., 1994; Økland et al., 1995; Youngston et al., 1998), differences in fitness traits and life history (Jonsson et al., 1991; Thodesen et al., 1999; McGinnity et al., 2003), and behavioral interactions between hatchery and wild juvenile salmon (Swain and Riddell, 1990; Johnsson et al., 1996; Clifford et al., 1998b). Much of this research has been driven by concerns about the impacts of escaped farmed Atlantic salmon in the North Atlantic Ocean (Hansen et al., 1991). Studies of the interactions between hatchery and wild salmonids, with few exceptions (see Myers et al., 2000), have primarily focused on interactions in freshwater environments. Interactions between hatchery and wild fishes in estuarine and marine environments have not received the same attention, but may have signifi- cant impacts on wild populations. To address this issue, we organized a symposium held at the 2003 annual meeting of the Western Division of the American Fisheries Society in San Diego, California. The session was titled Interactions of Hatchery and Wild Fishes in Marine and Estuarine Environments and included nine presentations. All but one presentation focused on salmonids. This Special Issue of Reviews in Fish Biology and Fisheries includes five papers that were submitted to the symposium.

Reviews in Fish Biology and Fisheries↗

Assessment of smolt condition for travel time analysis. Annual report 1988

Estimates of migration rates and travel times of juvenile salmonids within index reaches of the Columbia River basin are collected through the Smolt Monitoring Program for use by the Fish Passage Center. With increased reliance upon travel time estimates in 1988 by the Fish Passage Center, this study was implemented to monitor the biological attributes of juvenile chinook salmon Oncorhynchus tshawytscha and steelhead trout 0.- mykiss used for the travel time estimates, The physiological ability of fish to respond to stress was assessed by measuring levels of plasma cortisol, glucose, and chloride before and after a stress-challenge test. Most mid-Columbia and Snake river groups responded normally to the stress challenge exhibiting an increase in plasma glucose and cortisol and a slight decrease in chloride. Fish trucked to release sites were more stressed than those released directly from the hatchery, but most still responded to the stress challenge test normally. An abnormal or extreme stress response occurred when there were deviations from preferred protocol, disease problems at hatcheries, or when fish were trucked over long periods (7h). The development of smoltification was evaluated by measuring gill Na+K+-ATPase, plasma thyroxine, purines, and body morphology. Most groups were similar at the hatcheries but differed as the migration to McNary Dam proceeded. Gill ATPase activity increased 2-3 fold during the first 20 days of migration, after which it changed little. Fish with longer in-river travel times appeared to be more smolted than those which were in the river for a shorter period of time. The prevalence of bacterial kidney disease (BKD) in spring chinook salmon was evaluated using the enzyme linked immunosorbent assay (ELISA) and fluorescent antibody technique (FAT). Prevalence of BKD in groups tested using the ELISA method was as high as 99% at some downstream locations. A review of indices is presented as a guide, to the development of an index of smolt condition and preliminary data are presented. An index could be used as a tool to synthesize information on fish condition to assist with management and evaluation of the Water Budget.

Report↗

Avian disease at the Salton Sea

A review of existing records and the scientific literature was conducted for occurrences of avian diseases affecting free-ranging avifauna within the Salton Sea ecosystem. The period for evaluation was 1907 through 1999. Records of the U.S. Department of Agriculture, Bureau of Biological Survey and the scientific literature were the data sources for the period of 1907–1939. The narrative reports of the U.S. Fish and Wildlife Service's Sonny Bono National Wildlife Refuge Complex and the epizootic database of the U.S. Geological Survey's National Wildlife Health Center were the primary data sources for the remainder of the evaluation. The pattern of avian disease at the Salton Sea has changed greatly over time. Relative to past decades, there was a greater frequency of major outbreaks of avian disease at the Salton Sea during the 1990s than in previous decades, a greater variety of disease agents causing epizootics, and apparent chronic increases in the attrition of birds from disease. Avian mortality was high for about a decade beginning during the mid-1920s, diminished substantially by the 1940s and was at low to moderate levels until the 1990s when it reached the highest levels reported. Avian botulism ( Clostridium botulinum type C) was the only major cause of avian disease until 1979 when the first major epizootic of avian cholera ( Pasteurella multocidia ) was documented. Waterfowl and shorebirds were the primary species affected by avian botulism. A broader spectrum of species have been killed by avian cholera but waterfowl have suffered the greatest losses. Avian cholera reappeared in 1983 and has joined avian botulism as a recurring cause of avian mortality. In 1989, avian salmonellosis ( Salmonella typhimurium ) was first diagnosed as a major cause of avian disease within the Salton Sea ecosystem and has since reappeared several times, primarily among cattle egrets ( Bubulcus ibis ). The largest loss from a single epizootic occurred in 1992, when an estimated 155 000 birds, primarily eared grebes ( Podiceps nigricollis ), died from an undiagnosed cause. Reoccurrences of that unknown malady have continued to kill substantial numbers of eared grebes throughout the 1990s. The first major epizootic of type C avian botulism in fish-eating birds occurred in 1996 and killed large numbers of pelicans ( Pelecanus occidentalis & P . erythrorhynchos ). Avian botulism has remained as a major annual cause of disease in pelicans. In contrast, the chronic on-Sea occurrence of avian botulism in waterfowl and shorebirds of previous decades was seldom seen during the 1990s. Newcastle disease became the first viral disease to cause major bird losses at the Salton Sea when it appeared in the Mullet Island cormorant ( Phalacrocorax auritus) breeding colony during 1997 and again during 1998.

California↗