A review of potential pathogens of sea lice and the application of cleaner fish in biological control

There are many examples of successful biological control of pest populations in aquatic environments. This approach to sea louse control has environmental benefits and is cost-effective. The range of possible pathogens of lice is reviewed and epibionts recorded from sea lice, including the monogenean Udonella caligorum and ciliates, are examined. Baculoviruses when ingested by insects form occlusion bodies resulting in severe damage to the digestive system and subsequent death, and this may be a promising approach. Cleaner wrasse (Labridae) have been stocked commercially with farmed salmon since 1989, and recent work on improving the method is reviewed. Wrasse are sourced from a wild fishery and stocked at ratios of 1 to 25-150 salmon. Over 5 million wrasse are stocked annually in Norway and c 30% of smolts in Scotland were stocked with wrasse until 1998, when an outbreak of infections salmon anaemia (ISA) deterred many farmers from transferring wild fish to cages. A case study is given showing that salmon in cages stocked with wrasse had a burden of one to eight lice through the first year compared with up to 40 lice per fish on unprotected and untreated fish. Electivity indices were used to compare the relative composition of lice developmental stages on salmon in stocked and unstocked cages, and adult male and female lice were found to comprise only 6% of the population in cages with wrasse, compared with 49% adults on fish in control cages. Measures to improve the efficacy of wrasse as a way of cleaning salmon in the second production year include the use of refuges to assist over-wintering survival, and stocking ballan wrasse. Health hygiene includes sourcing wrasse in the farm locality, testing for pathogens, vaccination of wrasse and ultimately rearing wrasse for stocking. The role of wrasse in an IPM strategy is described.     

Wild salmonids and sea louse infestations on the west coast of Scotland: sources of infection and implications for the management of marine salmon farms

The sea louse (Lepeophtheirus salmonis Kr?yer) is a major health problem for both farmed and wild salmonids. This paper investigates louse epidemiology and management in the salmon-farming zone of western Scotland. Based on a review of the marine ecology of wild salmon (Salmo salar L) and sea trout (Salmo trutta L), and catch and farm production statistics, best estimates were made for numbers of wild and farm hosts present in coastal waters in March-June 2000. Applying data for ovigerous female louse infections and fecundity, the sources and risks of larval transmission to wild salmon and sea trout were modelled. Farm salmon in the second spring of production were the primary host group (98% of fish), while numbers of wild salmonids (< 1%) and escaped farm salmon (2%) were relatively insignificant. Farm salmon produced 97% of louse eggs at high levels (eight ovigerous lice per fish), and 78% at low levels (one per fish). Wild salmonids produced < 1% of eggs under both scenarios, but escaped farm salmon produced 3% and 21%, respectively. All hosts potentially cross-infect one another, but farm salmon are more likely to infect wild and farm smolts, and also other farm salmon. Monitoring of lice on sea trout in June 1998-2000 by the Association of West Coast Fisheries Trusts corroborated the model's conclusions. Localised epizootics occurred every year and coincided with the presence of ovigerous lice on local farms. In areas of mixed-year class production on farms, epizootics were evident every spring, but occurred every second spring in areas of single-year class production. In 1998-2000 at least 14-40% of sea trout were infected with potentially lethal infestations of lice. Ovigerous louse levels of < 0.005 per fish were required on farm salmon in the spring of 2000 to produce less eggs than those emitted by wild salmonids. With the industry's continued expansion, and thus increased numbers of farm salmon, a target of zero ovigerous lice will be required on farms to minimise impacts on wild salmonids. Due to the limited long-term efficacy and availability of louse medicines, management strategies are discussed which will improve control, including single-year class production over large areas, alternate S1-S1/2 smolt inputs, and 11-month production cycles.     

Semiochemical strategies for sea louse control: host location cues

The development of behavioural bioassays and electrophysiological recording techniques has enabled the role of semiochemicals to be investigated for the first time in Lepeophtheirus salmonis (Kr?yer). Adult male sea lice L salmonis were activated by, and attracted to, salmon-conditioned water (SCW) and SCW extract. Non-host fish odours, turbot-conditioned water (TCW) and turbot-conditioned water extract (TCW extract), elicited activation but no attraction in the lice. Solid-phase extraction techniques were developed to extract low molecular weight components of fish odours, and biological activity was shown to be confined to these. Electrophysiological activity from aesthetascs in the first antennae was found and neural responses to odours were recorded from the antennal nerves. Preliminary experiments on individual chemicals (isophorone, 1-octen-3-ol) linked to salmon revealed behavioural activation and electrophysiological responses in adult male L salmonis. Isophorone was shown to be significantly attractive to sea lice when placed in a slow-release system in a perfused tank of seawater in a choice situation. Proof of concept has been established for successfully extracting odour cues from seawater, analysing their biological activity and applying these to slow-release technologies for field trapping of lice. Future work involving linked GC-MS techniques using behaviour, electrophysiological responses, and organolepsis will establish further host location cues specific to Atlantic salmon.     

Temporal,environmental and management factors influencing the epidemiological patterns of sea lice (Lepeophtheirus salmonis) infestations on farmed Atlantic salmon (Salmo salar) in Scotland

In association with industry, an extensive database has been established on measurements of sea louse counts on farmed Atlantic salmon for the years 1996 to 2000 from 33 Scottish fish farms. These data include extensive counts on the sea louse species, Lepeophtheirus salmonis, at various stages of the life cycle and in particular the chalimus and mobile stages. There has been considerable speculation as to what factors might affect the abundance of sea lice, much of which is based on limited evidence. Our analyses show that there is tremendous variation in sea louse infestation patterns from year to year, whereas stock type, geographical region and coastal exposure do not appear to affect mean levels of abundance. In contrast, treatments lead to pronounced cycles of sea louse infestation with peaks and troughs at 3-week intervals, and these interventions are important if the sea louse levels on fish are to be controlled. There was no evidence of water temperature affecting the mean annual abundance of sea louse infestation.     

Towards selective breeding of Atlantic salmon for sea louse resistance: approaches to identify trait markers

Several species of sea lice, in particular Lepeophtheirus salmonis (Kr?yer), affect the welfare and condition of farmed salmon, with an estimated annual cost to the Scottish industry of 15-30 Pounds million. In Atlantic salmon, some stocks show resistance to L salmonis. Such natural resistance could be utilized for stock improvement using molecular genetic technologies. The development of molecular markers linked to resistance genes, allowing the identification of resistant fish, could increase the efficacy of selective breeding programmes. Various approaches to achieve this goal are described. One way to identify genes conferring resistance is to develop screens for salmon genes that are activated upon louse infection. One such screen--differential display--requires no previous knowledge of gene sequences, involves no preconceptions about which gene families are involved and can therefore identify novel genes. Preliminary results of comparative gene expression in sea-louse-challenged and control fish illustrate the application of differential display.     

Analysis and management of resistance to chemotherapeutants in salmon lice,Lepeophtheirus salmonis (Copepoda: Caligidae)

In Northern Europe and Canada, the salmon louse, Lepeophtheirus salmonis (Kr?yer), seriously affects the marine phase of salmon production. Although the problem is long-standing, the development of sustainable methods of pest management has been unable to keep pace with the intensification of production, leading to large-scale reliance on very few chemotherapeutants. This runs the risk of selecting for genetically determined resistance in target organisms. There are many examples of similar evolutionary adaptations in arthropod pests of arable crops, livestock and human health. Several hundred pest species are now documented as being resistant to one or more chemical classes of insecticides and acaricides. Many of these compounds are identical or closely related to ones currently employed against salmon lice. It is, therefore, opportune to consider what lessons have been learnt from contending with resistance in terrestrial organisms, the implications for sustainable use of chemotherapeutants in aquaculture, and the potential for developing effective resistance management strategies. An EU-funded project named SEARCH (QLK2-CT-2000-00809) has been initiated to explore in more detail the diagnosis, incidence, dynamics and management of resistance to chemotherapeutants in L salmonis.     

Observations on the metazoan parasites of the Atlantic salmon (Salmo salar) after its reintroduction into the Elbe River basin in the Czech Republic

In the years 1999-2002, first studies were carried out on the metazoan parasites of Atlantic salmon (Salmo salar L.) recently reintroduced into the Elbe River drainage system, after more than 50 years after the complete extinction of the Elbe salmon population. A total of six helminth species were recorded from salmon smolts from three streams of the Elbe River basin (Kamenice River, Jestedsky and Libocansky Brooks) in North Bohemia, Czech Republic, where S. salar fingerlings have been released since 1998: Gyrodactylus truttae Gl?ser, 1974, Crepidostomum metoecus Braun, 1900, Diplostomum spathaceum (Rudolphi, 1819) metacercariae, Raphidascaris acus (Bloch, 1799) adults and encapsulated larvae, Cystidicoloides ephemeridarum (Linstow, 1872), and Neoechinorhynchus rutili (Müller, 1780). Except for D. spathaceum, all these freshwater parasites have been received from the helminth fauna of the co-habiting brown trout (Salmo trutta fario L.). Due to local ecological conditions, the parasite faunae of both salmon and brown trout exhibited distinct qualitative and quantitative differences in the three localities. The finding of G. truttae on S. salar represents a new host record. Three helminth parasites of marine origin, the cestodes Eubothrium crassum (Bloch, 1779) and Scolex pleuronectis Müller, 1788 plerocercoids, and the nematode Anisakis simplex (Rudolphi, 1809) larvae were found in the single examined adult S. salar from the Kamenice River. New data on the geographical distribution of some nematode parasites of Salmo trutta fario L., Barbatula barbatula (L.) and Anguilla anguilla (L.) are presented.     

Using observed load distributions with a simple model to analyse the epidemiology of sea lice (Lepeophtheirus salmonis) on sea trout (Salmo trutta)

Sea lice are ectoparasites of salmonids that have been associated with the recent decline in sea trout numbers in north-west Europe. Observed patterns of louse load distribution between sea trout in the seas surrounding the UK, Ireland and Norway and a simple model have been used to analyse the epidemiology of lice. Loads are aggregated and deviate strongly from the Poisson distribution, although less than is observed with many other parasites. The louse numbers on fish from offshore sites are slightly less variable than for fish from coastal sites with comparable mean loads. Analysis of louse development stages and sexes shows that selection between hosts by sea lice plays a limited role. If host selection is absent, then associated poor condition would be caused by, not the cause of, high louse burdens; however the absence of such selection is not proved. Scenarios with infection that is patchy in space and time best generate the aggregated load patterns observed; these patches accord with observed swarms of copepodids. Prevalence patterns may indicate the movement of trout between environments. Control of copepodids in infection 'hot spots', either directly or through control of louse egg production in their catchment, may reduce louse loads on wild sea trout and, in particular, extreme and damaging loads.     

Molecular mechanisms and monitoring of permethrin resistance in human head lice

Head lice resistance to permethrin is mainly conferred by the knockdown resistance (kdr) trait, a voltage-sensitive sodium channel (VSSC) insensitivity factor. Three VSSC mutations (M815I, T917I and L920F) have been identified. Functional analysis of the mutations using the house fly VSSC expressed in Xenopus oocytes revealed that the permethrin sensitivity is reduced by the M827I (M815I) and L932F (L920F) mutations when expressed alone but virtually abolished by the T929I (T917I) mutation, either alone or in combination. Thus, the T917I mutation is primarily responsible for permethrin resistance in head lice. Comparison of the expression rates of channel variants indicates that the M815I mutation may play a role in rescuing the decreased expression of channels containing T917I. A step-wise resistance monitoring system has been established based on molecular resistance detection techniques. Quantitative sequencing (QS) has been developed to predict the VSSC mutation frequency in head lice at a population basis. The speed, simplicity and accuracy of QS made it an ideal candidate for a routine primary resistance monitoring tool to screen a large number of wild louse populations as an alternative to conventional bioassay. As a secondary monitoring method, real-time PASA (rtPASA) has been devised for more precise determination of low resistance allele frequencies. To obtain more detailed information on resistance allele zygosity, as well as allele frequency, serial invasive signal amplification reaction (SISAR) has been developed as an individual genotyping method. Our approach of using three tiers of molecular resistance detection should facilitate large-scale routine resistance monitoring of permethrin resistance in head lice using field-collected samples.     

Differential host utilisation by different life history stages of the fish ectoparasite Argulus foliaceus (Crustacea: Branchiura)

In this study we examine differences in the occurrence of life history stages of the destructive fish ectoparasite Argulus foliaceus (L., 1758) on eight fish species (stickleback, rudd, roach, gudgeon, bream, tench, crucian carp and common carp) sampled from a mixed-species recreational fishing lake on nine occasions during late spring and summer. Total numbers ofA. foliaceus, as well as the number of larval, juvenile and adult parasite stages, from each fish were recorded along with the fish species. Lice generally exhibited an aggregated distribution approximating a negative binomial distribution. Significant differences in the prevalence, intensity and intensity frequency distribution were observed between life history stages and between host species. In general, all life history stages of A. foliaceus exhibited an over-dispersed distribution. However, larval lice did show some degree of aggregation particularly within the stickleback samples. Infection data for parasite larval stages suggested that sticklebacks are more likely to be infected than other host species. For adult lice, however, carp appeared to be the main host. We propose that A. foliaceus infection characteristics are predominantly determined by the level of host exposure to the parasite and its life history stages (larval, juvenile and adult) rather than by an innate difference in host susceptibility related to individual host factors such as immune responses. We conclude that host exposure is determined by the parasite-host behavioural interplay related to species-specific ecology and behavioural traits such as microhabitat preference and normal swimming speed.     

Permethrin treatments in canadian forests. Part 1: Impact on stream fish

The effects of permethrin on native and caged fish when aerially applied directly to forest streams at dosages between 8.8 and 70 g ha^?1 were studied between 1976 and 1981. None of the applications caused mortality of caged or native fish in streams. Trout and salmon diets were altered by the treatments due to effects on fish food organisms. The duration of the effects varied from several months to over a year with increasing dosage. Reductions in salmonid growth rates and reductions in fish densities in treated areas, presumably due to emigration, were documented following severe impacts on aquatic invertebrates in salmon nursery streams. Growth rates and population densities both recovered within four months after treatment.     

Evidence for occurrence of an organophosphate-resistant type of acetylcholinesterase in strains of sea lice (Lepeophtheirus salmonis Krøyer)

Acetylcholinesterase (AChE) is the target of a major pesticide family, the organophosphates, which were extensively used as control agents of sea lice on farmed salmonids in the early 1990s. From the mid‐1990s the organophosphates dichlorvos and azamethiphos were seriously compromised by the development of resistance. AChE insensitive to organophosphate chemotherapeutants has been identified as a major resistance mechanism in numerous arthropod species, and in this study, target‐site resistance was confirmed in the crustacean Lepeophtheirus salmonis Krøyer isolated from several fish‐farming areas in Norway and Canada. A bimolecular rate assay demonstrated the presence of two AChE enzymes with different sensitivities towards azamethiphos, one that was rapidly inactivated and one that was very slowly inactivated. To our knowledge this is the first report of target‐site resistance towards organophosphates in a third class of arthropods, the Crustacea. Copyright © 2004 Society of Chemical Industry     

Estimating buffer zone widths for pesticide applications

A technique for estimating the width of buffer zones required around sensitive areas during pesticide applications has been devised and tested. The technique has been used to estimate the buffer width required around water bodies during ground-based permethrin applications in Canadian forests to prevent significant impact on fish and their food populations. A worst- case scenario was developed for environmental impact in water bodies result ing from ground-based permethrin applications, and a spray application was made under these worst-case conditions. Permethrin deposit on ground sheets was measured downwind of overlaid crosswind swaths. From these measurements the deposit at various downwind distances from a single crosswind swath was calculated, and a curvilinear regression line fitted to these values. Permethrin deposit downwind of multiple-swath applications was computed by adding the contributions from individual swaths. Mortality resulting from various permethrin concentrations was measured for Aedes aegypti larvae. Although these larvae are not an important food species for the fish species of interest, salmon and trout, they are more sensitive to permethrin than most aquatic invertebrates. Predicted mortality in populations of this species and Salmo gairdneri, rainbow trout, at various downwind distances from the permethrin application was calculated from the toxicological and spray-cloud dispersal data. Buffer width was estimated by choosing an acceptably low mortality, and determining the downwind distance at which this value was obtained. For example, a 20 m swath width was found to be adequate to limit mortality in A. aegypti andS. gairdneri populations to 10 and 0.1% during ground-based permethrin applications.     

Infection prevalence,seasonality and host specificity of actinosporean types (Myxozoa) in an Atlantic salmon fish farm located in Northern Scotland

A total of 28,387 oligochaetes belonging to the families Tubificidae, Lumbriculidae, Naididae and Enchytraeidae were examined at regular intervals for actinosporean infections from October 1996 to August 1998 from a freshwater salmon farm in Northern Scotland. A total of 21 types of actinosporeans belonging to seven collective groups synactinomyxon (three types), aurantiactinomyxon (four types), echinactinomyxon (five types), raabeia (six types), triactinomyxon (one type), neoactinomyxum (one type) and siedleckiella (one type) were found. Synactinomyxon type 1, echinactinomyxon type 1 and raabeia type 4 were most abundant. The overall infection prevalence of oligochaetes was 2.9%. Aurantiactinomyxon, synactinomyxon and neoactinomyxum were most common in summer and autumn. Raabeia was most common in spring and summer and echinactinomyxon in winter and spring. Siedleckiella was found only in spring and triactinomyxon in all seasons except winter. A positive relationship between water temperature and the number of actinosporean types released was observed. Most actinosporean types were found in only one host species.     

Occurrence of the microsporidian parasite Nucleospora salmonis in four species of salmonids from the Massif Central of France

Nucleospora salmonis (Hedrick, Groff et Baxa, 1991), an intranuclear microsporidian parasite of marine and freshwater fish, causes diseases mainly in salmonid species. Losses have been reported in stocks of salmonid fish reared in the region of Auvergne (France). The cause of chronic mortalities in the local host species raised in aquaculture and destined for supplementation of the river system Loire-Allier was examined. The presence of N. salmonis was confirmed by PCR and histology in Salmo salar L. previously and in newly investigated salmonid species, Salmo salar, Salmo trutta fario L., Thymallus thymallus (L.) and Salvelinus alpinus (L.), present in European streams. The infection by N. salmonis was consistent in all cases with characteristic symptoms of the disease in deceased or moribund fish. The small subunit ribosomal DNA from N. salmonis was partially sequenced and compared to previously characterised N. salmonis isolates. As a result, a genotype, or clonal entity, was attributed to N. salmonis among Atlantic salmon found along the Northern Atlantic coastal lines and other salmonid species co-inhabiting or co-cultivated in the Auvergne region.     

Medicines for sea lice

Sea louse (Family Caligidae: genera Caligus and Lepeophtheirus) infection of farmed salmonids represents a significant threat to animal welfare and undermines profitability. Lice may also act as vectors for the transmission of viral and bacterial pathogens. Pest-control programmes parallel those deployed in terrestrial livestock farming and include the use of parasiticides. The authorisation process for fish medicines varies widely between salmon farming countries and undue regulatory constraint may place farmers in one country at a competitive disadvantage. In many jurisdictions, fish are a 'minor' species and mounting demands for environmental assessment increase registration costs. A successful integrated louse-management strategy requires free access to a range of effective, chemically unrelated active ingredients deployed according to current best practice. Over-reliance on a limited number of products will lead, inevitably, to resistance, which is difficult to counter.     

Fumigation of potatoes with 2-aminobutane for control of gangrene and skin spot1

In Scotland 2-aminobutane (2-AB) has been used successfully for the control of gangrene (Phoma exigua var. foveata) and skin spot (Polyscytalum pustulans) in potatoes stored both in boxes and bulk. Gangrene and skin spot can be particularly troublesome in some areas of Scotland and under the prevalent harvesting conditions 2-AB provides a high level of control while other materials have been less effective.     

Development of vaccines against sea lice

A review of efforts to develop a vaccine against sea lice is presented together with analysis of the rationale behind the approaches and potential future directions. Vaccines against the caligid copepod, Lepeophtheirus salmonis, have the potential to be a cost-effective means of controlling the infection and avoid many of the disadvantages of medicine treatments. However, research towards such vaccines is in its infancy and approaches so far used have met with little or no success. Most strategies for sea louse vaccines have adopted methods used for vaccines against other ectoparasites. A vaccine against the cattle tick (Boophilus microplus) is in field use while other vaccines such as the sheep blowfly (Lucilia cuprina) vaccine are at an earlier stage of development. These haematophageous parasites ingest host antibody as part of a large blood meal which can target antigenic sites in the gut. However, the assumption that arachnid and insect physiology are directly comparable with that of sea lice is not proven, and this may partly explain the slow progress this approach has had with sea lice. Success in developing a louse vaccine will depend upon a better understanding of louse digestive biology, particularly an evaluation of whether the cattle tick model is applicable to the development of a louse vaccine. If the louse gut is to be targeted immunologically, critical antigens will need to be identified and evaluated, bearing in mind that an economic vaccine must include recombinant proteins or be a DNA vaccine. Alternatives to the louse gut as a target are also worth consideration. Antibodies could target critical host-parasite interactions that are amenable to disruption, although no such targets have been identified.     

Discharge consents in Scotland

The Scottish Environment Protection Agency (SEPA) is charged with the protection of the Scottish environment, and this is achieved through the regulation of polluting discharges and through consulting, influencing and educating others who interact with it. This paper describes aspects of the agency's regulatory work as it applies to the Scottish fish-farming industry. By far the largest sector of the industry in Scotland involves the rearing of fin-fish in cages, presently still dominated by Atlantic salmon, and the paper is based on experiences gained within this sector. The present circumstances affecting its development are described with reference to the environmental impacts associated with cage-rearing techniques used for production in marine waters. This paper briefly reviews the statutory background behind Scotland's system of discharge consents, including relevant aspects of European legislation. Methods developed to control the environmental risks posed by sea louse treatment chemicals are described. The concept of farming the sea is explored in relation to SEPA's 'allowable zone of effects' approach and the growing public concern about perceived environmental damage. Finally, the future prospects for the industry in Scotland are reviewed in relation to sea louse control.     

Residues of organochlorine insecticides and polychlorinated biphenyls in fish from Lakes Saint Clair and Erie, Canada--1968-76.

Eighteen species of fish from Lake Saint Clair and 19 species from Lake Erie were analyzed for organochlorine pesticides and polychlorinated biphenyls (PCBs) between 1968 and 1976. Mean residues of sigma DDT peaked at 1.19 ppm in longnose gar (Lepisosteus osseus) caught in Lake Saint Clair in 1970-71, but had declined in all species by 1975-76. Dieldrin levels in fish tissues increased over the same period. White bass (Morone chrysops), caught in 1975 in Lake Erie, had the highest mean residue of dieldrin at 0.17 ppm. PCB residues increased in some species and decreased in others. PCB residues exceeding the tolerance level of Health and Welfare Canada were found in the following: from Lake Saint Clair, smallmouth bass (Micropterus dolomieui) in 1975 and channel catfish (Ictalurus punctatus) in 1971; from Lake Erie, coho salmon (Oncorhynchus kisutch) in 1970, smallmouth bass, alewife (Alosa pseudoharengus), freshwater drum (Aplodinotus grunniens), and gizzard shad (Dorosoma cepedianum) in 1971, and white bass in 1971 and 1976.