Quantitative analysis of <Emphasis Type="Italic">Ichthyobodo salmonis</Emphasis> an ectoparasitic flagellate infecting juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> in hatcheries

Ichthyobodosis caused by the ectoparasitic flagellate Ichthyobodo salmonis is a significant cause of mortality in juvenile chum salmon Oncorhynchus keta reared in hatcheries of northern Japan. The present study established a real-time quantitative polymerase chain reaction assay (qPCR) of I. salmonis ribosomal DNA (rDNA) using SYBR Green. This assay allows monitoring of parasite infections for the epidemiological study and control of ichthyobodosis in hatcheries. qPCR showed high reproducibility for measurements between 1.0 and 1.0 × 10⁸ rDNA copy/μl. There was a significant positive correlation between the number of parasites and the amount of I. salmonis rDNA. A survey using the qPCR assay indicated that infection by I. salmonis was present in 23 of 87 hatcheries; parasite loads were estimated to be between 50 and 750 parasites/g juvenile body weight. These results demonstrate that our qPCR assay enables the surveying of juvenile chum salmon reared in hatcheries for infection by I. salmonis.     

Density‐dependent effects of eastern Kamchatka pink salmon (Oncorhynchus gorbuscha) and Japanese chum salmon (O. keta) on age‐specific growth of western Alaska chum salmon

Hatcheries release >4.5 billion juvenile Pacific salmon (Oncorhynchus spp.) into the North Pacific Ocean annually, raising concerns about competition with wild salmon populations. We used retrospective scale analysis to investigate how the growth of chum salmon (O. keta) from western Alaska is affected by the abundance of chum salmon from Japanese hatcheries and wild pink salmon (O. gorbuscha) from the Russian Far East. Over nearly five decades, the growth of Kuskokwim River chum salmon was negatively correlated with the abundance of Japanese hatchery chum salmon after accounting for the effects of sex and spring/summer sea‐surface temperature in the Bering Sea. An effect of wild eastern Kamchatka pink salmon abundance on the growth of Kuskokwim River salmon was detectable but modest compared to the intraspecific competitive effect. A decrease in Japanese hatchery chum salmon releases in 2011–2013 was not associated with increased growth of Bering Sea chum salmon. However, the abundance of wild chum salmon from the Russian Far East increased during that time, possibly obscuring reduced competition with hatchery chum salmon. Our results support previous evidence that chum salmon are affected by intraspecific competition, and to a lesser extent interspecific competition, in the North Pacific, underscoring that the effects of salmon hatchery production transcend national boundaries.     

Changing salmon: An analysis of body mass,abundance, survival,and productivity trends across 45 years in Puget Sound

Pacific salmon and trout (Oncorhynchus spp., Salmonidae) of the Puget Sound region of Washington State, USA, have experienced recent and longer‐term (multidecadal) variability in abundance while supporting robust fisheries. As part of the post‐season salmon management process, population‐specific estimates of total adult abundance to Puget Sound (Strait of Juan de Fuca) for pink (O. gorbuscha), chum (O. keta), coho (O. kisutch), sockeye (O. nerka), and Chinook (O. tshawytscha) salmon and steelhead trout (O. mykiss) are calculated annually. We compiled annual estimates of body mass, abundance and survival of hatchery‐ and naturally produced salmon from 1970 to 2015 to compare spatial and temporal patterns across species. Average weights of adult salmon and steelhead returning to Puget Sound, with the exception of coho salmon, have decreased since the 1970s. Temporal trends in abundance, survival and productivity varied by species and origin (hatchery vs. naturally produced). Generally, abundance and survival rates of natural‐origin species decreased whereas those of hatchery‐produced species did not, which is in contrast with other studies' general conclusions of decreasing survival among Puget Sound salmonids. Species diversity has decreased in recent years, with salmonids that rely on a short freshwater rearing phase in the natural environment (hatchery‐produced fish and naturally produced pink and chum) representing >90% of total returns in most years. This new information reveals patterns of body size, abundance, survival and productivity across species, life history and rearing type over the past 45 years and, in doing so, demonstrates the strength in multidecadal, multifactor time series to critically evaluate salmonid species.     

Ultraviolet irradiation is an effective alternative to ozonation as a sea water treatment to prevent Kudoa neurophila (Myxozoa: Myxosporea) infection of striped trumpeter,Latris lineata (Forster)

Myxozoan parasites are known pathogens of cultured finfish. Kudoa neurophila n. comb. (Grossel, Dyková, Handlinger & Munday) has historically infected hatchery‐produced striped trumpeter, Latris lineata (Forster in Bloch and Schneider), a candidate species for seacage aquaculture in Australia. We examined the efficacy of four water treatment methods to prevent K. neurophila infection in post‐larval (paperfish) and juvenile striped trumpeter. Treatments included dose‐controlled ultraviolet irradiation [hydro‐optic disinfection (HOD)], ozone with conventional UV (ozone), mechanical filtration at 25 μm and then foam fractionation (primary filtration), and 50‐μm‐filtered sea water (control). In post‐larvae (initially 10.3 ± 2.7 g, mean ± SD, 259 days post‐hatching, dph), the infection prevalence (PCR test) after 51 days was 93 ± 12% in the control, 100 ± 0% in primary filtration and 0 ± 0% in both ozone and HOD. Likewise, in juveniles (initially 114 ± 18 g, 428 dph), prevalence was 100 ± 0% in the control and primary filtration treatments with no infection detected in ozone and HOD. Concurrently, there was a 50–100% reduction in heterotrophic bacteria and 100% reduction in presumptive Vibrio sp. in sea water HOD and ozone treatments. HOD with a dose of ≥44 mJ cm^?2 UV was as effective as ozonation at >700 mV ORP for 10 min, in preventing K. neurophila infection.     

Comprehensive antibiotic susceptibility profiling of Chilean Piscirickettsia salmonis field isolates

Antibiotics have been extensively used against infections produced by Piscirickettsia salmonis, a fish pathogen and causative agent of piscirickettsiosis and one of the major concerns for the Chilean salmon industry. Therefore, the emergence of resistant phenotypes is to be expected. With the aim of obtaining a landscape of the antimicrobial resistance of P. salmonis in Chile, the susceptibility profiles for quinolones, florfenicol and oxytetracycline (OTC) of 292 field isolates derived from main rearing areas, different hosts and collected over 5 years were assessed. The results allowed for the determination of epidemiological cut‐off values that were used to characterize the pathogen population. This work represents the first large‐scale field study addressing the antimicrobial susceptibility of P. salmonis, providing evidence of the existence of resistant types with a high incidence of resistance to quinolones. Remarkably, despite the amounts and frequency of therapies, our results disclosed that the issue of resistance to florfenicol and OTC is still in the onset.     

Early‐stage sea lice recruits on Atlantic salmon are freshwater sensitive

Sea lice are significant parasites of marine and brackish farmed fishes. Freshwater bathing is a potential control option against numerous sea lice species, although has been viewed as futile against those that are capable of tolerating freshwater for extended periods. By comparing freshwater survival times across host‐attached stages of Lepeophtheirus salmonis (Krøyer), a key parasite in Atlantic salmon farming, we show the first attached (copepodid) stage undergoes 96–100% mortality after 1 h in freshwater, whereas later attached stages can tolerate up to 8 days. Thus, regular freshwater bathing methods targeting the more susceptible attached copepodid stage may successfully treat against L. salmonis and potentially other sea lice on fish cultured in marine and brackish waters.     

Development of a Piscirickettsia salmonis immersion challenge model to investigate the comparative susceptibility of three salmon species

Piscirickettsia salmonis, the aetiological agent of salmonid rickettsial septicaemia (SRS), is a global pathogen of wild and cultured marine salmonids. Here, we describe the development and application of a reproducible, standardized immersion challenge model to induce clinical SRS in juvenile pink (Oncorhynchus gorbuscha), Atlantic (Salmo salar) and sockeye salmon (O. nerka). Following a 1‐hr immersion in 10⁵ colony‐forming units/ml, cumulative mortality in Atlantic salmon was 63.2% while mortality in sockeye salmon was 10%. Prevalence and levels of the bacterium in kidney prior to onset of mortality were lower in sockeye compared with Atlantic or pink salmon. The timing and magnitude of bacterial shedding were estimated from water samples collected during the exposure trials. Shedding was estimated to be 82‐fold higher in Atlantic salmon as compared to sockeye salmon and peaked in the Atlantic salmon trial at 36 d post‐immersion. These data suggest sockeye salmon are less susceptible to P. salmonis than Atlantic or pink salmon. Finally, skin lesions were observed on infected fish during all trials, often in the absence of detectable infection in kidney. As a result, we hypothesize that skin is the primary point of entry for P. salmonis during the immersion challenge.     

Experimental infections with Lepeophtheirus salmonis (Krøyer) on threespine sticklebacks, Gasterosteus aculeatus L., and juvenile Pacific salmon, Oncorhynchus spp

Experimental infections with Lepeophtheirus salmonis (Krøyer) were established on threespine sticklebacks, Gasterosteus aculeatus L., juvenile pink, Oncorhynchus gorbuscha (Walbaum), and chum, Oncorhynchus keta (Walbaum), salmon. The prevalence and abundance of infections were initially higher on sticklebacks than on either salmon species. The initial prevalence and intensity of infections on chum salmon were higher than those on pink salmon, and declined on both species during louse development. The rate of parasite development to adult stages was similar on all species although development beyond the preadult stage was not observed on sticklebacks. These results confirm previous field observations on the occurrence and development of L. salmonis on threespine sticklebacks.     

Prevalence,geographic distribution and phenotypic differences of Piscirickettsia salmonis EM‐90‐like isolates

Early reports accounted for two main genotypes of Piscirickettsia salmonis, a fish pathogen and causative agent of piscirickettsiosis, placing the single isolate EM‐90 apart from the prototypic LF‐89 and related isolates. In this study, we provide evidence that, contrary to what has been supposed, the EM‐90‐like isolates are highly prevalent and disseminated across Chilean marine farms. Molecular analysis of 507 P. salmonis field isolates derived from main rearing areas, diverse hosts and collected over 6 years, revealed that nearly 50% of the entire collection were indeed typed as EM‐90‐like. Interestingly, these isolates showed a marked host preference, being recovered exclusively from Atlantic salmon (Salmo salar) samples. Although both strains produce undistinguishable pathological outcomes, differences regarding growth kinetics and susceptibility to the antibiotics and bactericidal action of serum could be identified. In sum, our results allow to conclude that the EM‐90‐like isolates represent an epidemiologically relevant group in the current situation of piscirickettsiosis. Based on the consistency between genotype and phenotype exhibited by this strain, we point out the need for genotypic studies that may be as important for the Chilean salmon industry as the continuous surveillance of antimicrobial susceptibility patterns.     

Effect of treatment of chum salmon Oncorhynchus keta (Walbaum) eggs with 1,3;1,6‐β‐D‐glucans on their development and susceptibility to Saprolegnia infection

The effects of six 1,3;1,6‐β‐D‐glucooligo‐ and polysaccharides with different structures (ranging from 1 to 10 kDa in molecular mass and containing 10–25% of β‐1,6‐linked glucose residues) from brown algae, Saccharina cichorioides, on development of the chum salmon, Oncorhynchus keta (Walbaum), were evaluated. Exposure of chum salmon eggs to 1,3;1,6‐β‐D‐glucans with a molecular mass of more than 2 kDa increased the survival of embryos and juveniles and their resistance to Saprolegnia infection by up to 2.5‐fold, leading to a weight gain in juveniles of 40–55% compared with The control chum salmons. The 1,3;1,6‐β‐D‐glucans with molecular mass of 6–8 kDa and used at a at concentration of 0.5 mg mL^?1 rendered the best stimulative effect.     

Flavobacteria colonizing the early life stages of hatchery‐incubated Chinook salmon Oncorhynchus tshawytscha (Walbaum 1792) are markedly diverse

Flavobacterial diseases are significant impediments to hatchery‐based fishery conservation and aquaculture productivity worldwide. Recent studies revealed a multitude of novel flavobacteria within the reproductive fluids and unfertilized eggs of feral Chinook salmon Oncorhynchus tshawytscha broodstock, some of which were associated with systemic disease. Herein, embryonated eggs/fry from these broodstock were assayed for flavobacteria while in incubator stacks in three hatcheries over 2 years, as was the water entering hatchery incubators. Overall, >65% of sampled eggs and 38% of fry were colonized by flavobacteria. One hundred and ninety‐one egg and fry‐associated flavobacterial isolates were characterized phenotypically and via 16S rRNA gene sequencing and phylogenetic analyses, revealing that the majority fell into 22 clades (i.e., 15 Flavobacterium spp. groups and seven Chryseobacterium spp. groups) that varied in presence by facility. Although some matched previously described fish‐pathogenic species, the majority were distinct from all described flavobacteria and likely represent novel species. Of concern, iodophor disinfection at the commonly utilized dose/duration for egg‐surface disinfection did not eliminate flavobacteria. Results also implicated maternal routes of infection and source water for some flavobacteria. In total, study findings underscore the complexity of flavobacterial ecology within hatchery environments and highlight the need for improved hatchery biosecurity practices.     

Aeromonas salmonicida infection levels in pre‐ and post‐stocked cleaner fish assessed by culture and an amended qPCR assay

Due to increasing resistance to chemical therapeutants, the use of ‘cleaner fish’ (primarily wrasse, Labridae, species) has become popular in European salmon farming for biocontrol of the salmon louse, Lepeophtheirus salmonis (Krøyer). While being efficient de‐licers, cleaner fish mortality levels in salmon cages are commonly high, and systemic bacterial infections constitute a major problem. Atypical furunculosis, caused by Aeromonas salmonicida A‐layer types V and VI, is among the most common diagnoses reached in clinical investigations. A previously described real‐time PCR (qPCR), targeting the A. salmonicida A‐layer gene (vapA), was modified and validated for specific and sensitive detection of all presently recognized A‐layer types of this bacterium. Before stocking and during episodes of increased mortality in salmon cages, cleaner fish (primarily wild‐caught wrasse) were sampled and screened for A. salmonicida by qPCR and culture. Culture indicated that systemic bacterial infections are mainly contracted after salmon farm stocking, and qPCR revealed A. salmonicida prevalences of approximately 4% and 68% in pre‐ and post‐stocked cleaner fish, respectively. This underpins A. salmonicida's relevance as a contributing factor to cleaner fish mortality and emphasizes the need for implementation of preventive measures (e.g. vaccination) if current levels of cleaner fish use are to be continued or expanded.     

The placement of Gyrodactylus salmonis (Yin & Sproston) in the molecular phylogeny of studied members of the Gyrodactylus wageneri‐group parasitizing salmonids

The molecular phylogeny of Gyrodactylus salmonis from brook charr, Salvelinus fontinalis, rainbow trout, Oncorhynchus mykiss, cutthroat trout, O. clarkii, and Atlantic salmon, Salmo salar, in Canada is presented using sequences from ITS‐rDNA and the mitochondrial COX 1 gene. Sequence variation among G. salmonis specimens from the different North American hosts was consistent with within‐species variation reported for other Gyrodactylus. Sequence data are compared to those from other members of the wageneri group parasitizing salmoniform fishes in northern Europe (G. derjavini, G. derjavinoides, G. lavareti, G. salaris, G. salvelini, G. teuchis and G. truttae) and Asia (G. brachymystacis). Sequence divergence between G. salmonis and the recently described G. salvelini on Arctic charr, Salvelinus alpinus, in Finland was consistent with within‐species levels of variation in Gyrodactylus; however, phylogenetic analyses and morphological comparisons provided evidence of their distinctiveness such that they appear to be sister species. They grouped with G. lavareti (a parasite of a coregonid) to form a clade separate from European and Asian species of the wageneri lineage known from salmonid fish. Further study of gyrodactylids from across salmonid, coregonid and thymallid fish in the northern hemisphere would shed more light on the phylogeography of these parasites and serves as an important backdrop to understanding the evolution of their emergent virulence.     

Substrate type and brightness preference of Schizothorax wangchiachii and Percocypris pingi juveniles

It is advantageous for fish to choose different substrate types and brightness based on their current life cycle stage, foraging requirements, predator avoidance and water conditions. We determined the substrate type and brightness preference of hatchery‐reared and wild juvenile Schizothorax wangchiachii and Percocypris pingi under laboratory conditions. Individuals and groups were exposed to four kinds of substrate types (sand: diameter < 1 mm, small gravel: diameter approximately 2–3 cm, large gravel: diameter approximately 10–15 cm, sand and medium gravel mix: diameter approximately 4–7 cm) and two kinds of substrate brightness (black and white). The results showed that hatchery‐reared and wild S. wangchiachii and P. pingi significantly preferred large gravel and black substrates regardless of the number of fish (p < .05). P. pingi had a significantly higher preference for black substrate than S. wangchiachii both in hatchery‐reared and wild individuals (p < .05). Hatchery‐reared and wild individuals of the two species shared a preference for large gravel and black substrates suggesting that substrate preferences might be genetically based. The findings in the present study could be used to improve the rearing conditions for the two species in hatchery and thereafter to enhance its adaptability after releasing to the wild.     

Prey partitioning and use of insects by juvenile sockeye salmon and a potential competitor,threespine stickleback,in Afognak Lake,Alaska

Freshwater growth of juvenile sockeye salmon (Oncorhynchus nerka) depends upon the quality and quantity of prey and interactions with potential competitors in the foraging environment. To a large extent, knowledge about the ecology of lake‐rearing juvenile sockeye salmon has emerged from studies of commercially important runs returning to deep nursery lakes, yet information from shallow nursery lakes (mean depth ≤ 10 m) is limited. We examined seasonal and ontogenetic variation in diets of juvenile sockeye salmon (N = 219, 30–85 mm) and an abundant potential competitor, threespine stickleback (Gasterosteus aculeatus; N = 198, 42–67 mm), to understand their foraging ecology and potential trophic interactions in a shallow Alaska lake. This study revealed that adult insects made up 74% of all sockeye salmon diets by weight and were present in 98% of all stomachs in Afognak Lake during the summer of 2013. Diets varied temporally for all fishes, but small sockeye salmon (<60 mm) showed a distinct shift in consumption from zooplankton in early summer to adult insects in late summer. We found significant differences in diet composition between sockeye salmon and threespine stickleback and the origin of their prey indicated that they also separated their use of habitat on a fine scale; however, the two species showed overlap in size selectivity of zooplankton prey. Considering that aquatic insects can be a primary resource for juvenile sockeye salmon in Afognak Lake, we encourage the development of nursery lake carrying capacity models that include aquatic insects as a prey source for sockeye salmon.     

Impacts of parasites on marine survival of Atlantic salmon: a meta‐analysis

Parasites can, in theory, have large impacts on the survival of fish populations. One method to evaluate such impacts on anadromous species is to apply manipulative field experiments in which parallel groups of antiparasitically treated and non‐treated fish are simultaneously released and then subsequently recaptured as returning adults. A systematic review and meta‐analysis on all such Norwegian studies on Salmo salar provided a data set for the time period 1996 to 2011 on 118 release groups comprising 657 624 fish released and 3989 recaptured. The overall risk ratio (RR) was estimated to be 1.18 (95% CI: 1.07–1.30). The effect varied strongly between groups, (Higgins I² = 40.1%). Over 70% of this heterogeneity could be explained by the release location, time period and baseline survival. The most important predictor variable was baseline survival. In groups with low recapture in the control group (low baseline survival), the effect of treatment was high (RR = 1.7), while in groups with high recapture in the control group (high baseline survival), there was no effect of treatment (RR ~ 1.00). The most prevalent parasite in the region affected by the drugs administered was Lepeophtheirus salmonis. Hence, the meta‐analysis supports the hypothesis that L. salmonis contributes to the mortality of S. salar during outward migration. However, the effect of treatment was not consistent, but was evidently strongly modulated by other risk factors. The results suggest that the population‐level effects of parasites cannot be estimated independently of other factors affecting the marine survival of S. salar.     

The Three‐spined Stickleback,Gasterosteus aculeatus Linnaeus 1758, plays a minor role as a host of Lepeophtheirus salmonis (Krøyer 1837) in the Gulf of Maine

The sea louse, Lepeophtheirus salmonis (Krøyer 1837), is a significant parasite of farmed salmon throughout the Northern Hemisphere. Management of on‐farm louse populations can be improved by understanding the role that wild fish play in sustaining and providing refuge for the local population of sea lice. In this study, 1,064 sticklebacks were captured. Of these animals, 176 individuals were carrying a total of 238 sea lice, yielding a prevalence and intensity of 16.5% and 1.4 lice per fish, respectively. Detailed examination of the sea lice on the three‐spined sticklebacks captured in Cobscook Bay found two L. salmonis individuals using three‐spined sticklebacks as hosts. A 2012 survey of wild fish in Cobscook Bay, Maine, found multiple wild hosts for Caligus elongatus (von Nordmann 1832), including three‐spined sticklebacks (Gasterosteus aculeatus L.), but no L. salmonis were found in this earlier study.     

Piscirickettsia salmonis infection in cultured lumpfish (Cyclopterus lumpus L.)

A Piscirickettsia salmonis infection was diagnosed in lumpfish (Cyclopterus lumpus L.) juveniles held in a marine research facility on the west coast of Ireland. The main clinical signs and pathology included marked ascites, severe multifocal liver necrosis and severe diffuse inflammation and necrosis of the exocrine pancreas and peri‐pancreatic adipose tissue. Numerous Piscirickettsia‐like organisms were observed by histopathology in the affected organs, and the bacterial species was characterized by molecular analysis. Sequencing of the partial 16S rDNA gene and internal transcribed spacer region showed the lumpfish sequences to be closely related to previously identified Atlantic salmon (Salmo salar L.) sequences from Ireland. To the authors’ knowledge, this is the first detection of P. salmonis in lumpfish worldwide. The infection is considered potentially significant in terms of lumpfish health and biosecurity.