Polyphasic characterization of Aeromonas salmonicida isolates recovered from salmonid and non‐salmonid fish

Michigan's fisheries rely primarily upon the hatchery propagation of salmonid fish for release in public waters. One limitation on the success of these efforts is the presence of bacterial pathogens, including Aeromonas salmonicida, the causative agent of furunculosis. This study was undertaken to determine the prevalence of A. salmonicida in Michigan fish, as well as to determine whether biochemical or gene sequence variability exists among Michigan isolates. A total of 2202 wild, feral and hatchery‐propagated fish from Michigan were examined for the presence of A. salmonicida. The examined fish included Chinook salmon, Oncorhynchus tshawytscha (Walbaum), coho salmon, O. kisutcha (Walbaum), steelhead trout, O. mykiss (Walbaum), Atlantic salmon, Salmo salar L., brook trout, Salvelinus fontinalis (Mitchill), and yellow perch, Perca flavescens (Mitchill). Among these, 234 fish yielded a brown pigment‐producing bacterium that was presumptively identified as A. salmonicida. Further phenotypic and phylogenetic analyses identified representative isolates as Aeromonas salmonicida subsp. salmonicida and revealed some genetic and biochemical variability. Logistic regression analyses showed that infection prevalence varied according to fish species/strain, year and gender, whereby Chinook salmon and females had the highest infection prevalence. Moreover, this pathogen was found in six fish species from eight sites, demonstrating its widespread nature within Michigan.     

Stress-induced impairment of predator evasion and non-predator mortality in Pacific salmon

This study examined whether rapid recovery from stress-induced impairment in predator evasion, observed in previous studies on coho salmon, Oncorhynchus kisutch (Walbaum), was a general characteristic of different stocks of the same species and different species of Pacific salmon. We monitored stress-induced non-predator mortality, predator evasion and Cortisol concentrations of smolts of coho and spring Chinook, O. tshawytscha (Walbaum), after administration of standardized single and multiple handling stresses. Marked differences in the response to handling stress among stocks of coho and spring chinook smolts were evident, with recovery from impaired predator evasion occurring within 24 h of a 30-s handling stress for coho smolts and 24 h of a 1-min handling stress for spring chinook smolts. Differences in stress-induced non-predator mortality among stocks were also observed. The results point to the importance of screening hatchery salmonid stocks to assess differing capabilities of dealing with stress.     

Impact of bacteria on short-term storage of salmonid eggs

Unfertilized eggs of steelhead, Oncorhynchus mykiss (Walbaum 1792), were stored at a low temperature (～0°C) with antibiotics (penicillin and streptomycin) for up to 22 days and fertility assessed. Unfertilized eggs of chinook salmon, Oncorhynchus tshawytscha (Walbaum 1792), were stored at a low temperature (～0°C) either with or without antibiotics for up to 24 days and fertility assessed. There were no significant decreases in fertility under these conditions. Steelhead eggs were maintained at 4°C for up to 15 days with or without added bacteria and with or without gentamicin. Gentamicin inhibited bacterial growth during storage at 4°C and its use resulted in a less pronounced decline in fertility with storage. These results suggest that low temperatures and antibiotics play an important role in extending the period over which salmonid eggs can be successfully stored.     

Comparison of histopathology caused by Vibrio anguillarum and Vibrio ordalii in three species of Pacific salmon*

Abstract. The histopathology associated with naturally acquired vibriosis in chum salmon, Oncorhynchus keta (Walbaum), fingerlings caused by Vibrio anguillarum was compared with that caused by infection with Vibrio ordalii. Pathogenesis of the two forms was found to be different. Bacteraemia caused by V. anguillarum occurred in the early stages with pronounced histopathological changes in blood, loose connective tissue, kidney, spleen, gills and posterior gastrointestinal tract. Bacterial cells appeared uniformly dispersed throughout the affected tissues but were most abundant in the blood. With V. ordalii, bacteraemia developed only in late stages of the disease and the concentration of bacterial cells per ml of blood was less than in the V. anguillarum infection by a factor of 10²–10³. Tissues with most pronounced changes were skeletal and cardiac muscle, anterior and posterior gastrointestinal tract and the gills. Vibrio ordalii observed in the tissues was not evenly dispersed but was present within tissue as colonies or aggregates of cells. The differences in pathology observed in naturally infected chum salmon were produced experimentally with each pathogen by waterborne exposure of chum; coho, Oncorhynchus kisutch (Walbaum); and chinook salmon, Oncorhynchus tshawytscha (Walbaum). Severe decreases in circulating leucocytes accompanied bacteraemia caused by either bacterial species.     

Molecular testing of adult Pacific salmon and trout (Oncorhynchus spp.) for several RNA viruses demonstrates widespread distribution of piscine orthoreovirus in Alaska and Washington

This research was initiated in conjunction with a systematic, multiagency surveillance effort in the United States (U.S.) in response to reported findings of infectious salmon anaemia virus (ISAV) RNA in British Columbia, Canada. In the systematic surveillance study reported in a companion paper, tissues from various salmonids taken from Washington and Alaska were surveyed for ISAV RNA using the U.S.‐approved diagnostic method, and samples were released for use in this present study only after testing negative. Here, we tested a subset of these samples for ISAV RNA with three additional published molecular assays, as well as for RNA from salmonid alphavirus (SAV), piscine myocarditis virus (PMCV) and piscine orthoreovirus (PRV). All samples (n = 2,252; 121 stock cohorts) tested negative for RNA from ISAV, PMCV, and SAV. In contrast, there were 25 stock cohorts from Washington and Alaska that had one or more individuals test positive for PRV RNA; prevalence within stocks varied and ranged from 2% to 73%. The overall prevalence of PRV RNA‐positive individuals across the study was 3.4% (77 of 2,252 fish tested). Findings of PRV RNA were most common in coho (Oncorhynchus kisutch Walbaum) and Chinook (O. tshawytscha Walbaum) salmon.     

Effect of Dietary Inclusion of N-Acetyl Cysteine on Mucus Viscosity and Susceptibility of Rainbow Trout, Oncorhynchus mykiss, and Atlantic Salmon, Salmo salar, to Amoebic Gill Disease

The treatment of amoebic gill disease (AGD) in cultured Atlantic salmon, Salmo salar L., using mucolytic agents has been previously reported. The agent L‐cysteine ethyl ester reduces salmonid mucus viscosity and potentially increases the flushing of the gill. In the present study, the effects of the mucolytic agent N‐acetyl cysteine (NAC) were assessed. Cutaneous mucus from rainbow trout, Oncorhynchus mykiss Walbaum, and Atlantic salmon was shown to have reduced viscosity when mixed in vitro with 100 or 200 μg/mL NAC. Saltwater‐acclimated rainbow trout and Atlantic salmon were fed an oil‐incorporated, NAC‐medicated diet (8 g NAC/kg diet) for up to 24 d and challenged with inoculation of 300 cells/L Neoparamoeba spp., the etiological agent of AGD. Control fish were fed normal oil‐coated pellets and received no NAC. NAC medication failed to reduce the severity of gill lesions associated with AGD even though the mucus viscosity from medicated fish was less than that of controls. Oral NAC medication does not appear to be an effective method for controlling AGD in salmonids despite reducing cutaneous mucus viscosity.     

Distribution and feeding of juvenile Pacific salmon in freshwater tidal creeks of the lower Fraser River, British Columbia

Abstract This study examined juvenile salmonid use of a freshwater tidal creek system draining a wetland on the floodplain of the lower Fraser River, British Columbia, Canada. Chum, Oncorhynchus keta (Walbaum), chinook, O. tshawytscha (Walbaum), and sockeye, O. nerka (Walbaum), salmon fry were abundant in the tidal creeks in spring. The fry were found in non-natal habitat up to 1.5 km from the main channel of the river. The salmon fry ate dipteran adults, larvae and pupae, cyclopoid and harpacticoid copepods, and Collembola. Mysids Neomysis mercedis Holmes (Walbaum), and amphipods, Crangonyx richmondensis occidentalis (Hubricht and Harrison), were also consumed. The upper reaches of an undisturbed creek were the winter rearing habitat for presmolt coho salmon, O. kisutch (Walbaum), where this species ate dipteran pupae and larvae as well as a freshwater isopod, Asellus communis Say (Walbaum).     

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.     

Salmonid colonization of new streams in Glacier Bay National Park, Alaska

Abstract. Following the rapid recession of a neoglacial ice sheet within the last 250 years, colonization of recently deglaciated streams by salmonid fishes was investigated in Glacier Bay National Park, south-eastern Alaska. The primary factors governing the establishment, species diversity composition and abundance of salmonids in Glacier Bay streams were water temperature, sediment loading and stream discharge. No salmonids were found in the turbid meltwater streams emerging from retreating ice. Coho, Oncorhynchus kisutch (Walbaum), and sockeye, Oncorhynchus nerka (Walbaum). salmon and Dolly Vardcn, Salvelinus malma (Walbaum), charr were the first salmonids to colonize the youngest clearwater stream. Juvenile Dolly Varden were more abundant than juvenile coho salmon in the most recently formed clearwater stream because of the characteristic absence of pool habital. Densities of juvenile coho salmon were six times greater in a stream with a series of lakes compared with a stream of similar age without lakes. Future advancement of salmonid stocks will probably depend upon the rate and extent of the development of riparian vegetation and inputs of large woody debris from the developing forest to provide further instream cover, habitat variation and channel stabilization.     

Mortality in juvenile salmonids passed through an agricultural Hidrostal pump

Abstract Hidrostal pumps have been successfully employed in live fish transport, yet their effectiveness in fish passage is incompletely understood. This study investigates juvenile salmonid mortality in experimental passage trials through Hidrostal pumps at an agricultural pump facility in Washington State, USA. The effects of impeller pitch, rotational speed and fish body size on passage survival were examined. Hatchery‐reared salmonids [Oncorhynchus mykiss (Walbaum) and Oncorhynchus kisutch (Walbaum)] were introduced to low‐ and high‐pitch impeller assemblies operating at two speeds. Instantaneous mortality rates ranged from 0 to 4% for high‐pitch trials and from 3 to 10% for low‐pitch trials. Larger fish experienced sublethal injury at higher rates (approximately 60% injured) than smaller fish (approximately 23% injured) and exhibited greater susceptibility to injury at higher pump speed. Injury between trials was compared by ranking according to severity and summed for each treatment; greater injury severity was found for the low‐pitch impeller and from higher rotational speeds. Although injury and mortality rates to fish passing through Hidrostal pumps may be reduced through the use of higher‐pitch impellers and lower operational speeds, the use of pump‐bypass facilities may be warranted where acceptable impact thresholds are low.     

Ceratomyxa shasta genotypes cause differential mortality in their salmonid hosts

Ceratomyxa shasta is a myxozoan parasite of salmonid fish. In natural communities, distinct genotypes of the parasite are associated with different salmonid hosts. To test the hypothesis that genotypes of C. shasta cause differential mortality, the polychaete host was experimentally infected with different parasite genotypes. Genotype I was obtained from Chinook salmon, Oncorhynchus tshawytscha, and genotype II from either coho salmon, O. kisutch, or rainbow trout, O. mykiss, We then challenged four salmonid strains: Chinook and coho salmon that occur in sympatry with the parasite and allopatric Chinook salmon and rainbow trout. Parasite genotype I caused mortality only in Chinook strains, although mortality in the allopatric strain also occurred from exposure to genotype II. A second experiment demonstrated that genotype II could be separated into two biotypes based on differential mortality in rainbow trout and coho salmon. These differential patterns of mortality as a result of infection by certain genotypes of C. shasta support field observations and suggest a co‐evolutionary relationship between these parasites and their hosts.     

Genetic analysis of lysozyme activity and resistance to vibriosis in farmed Chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Twelve full-sib families of chinook salmon, Oncorhynchus tshawytscha (Walbaum), reared in two environments, were examined for differences in disease resistance following a natural outbreak of vibriosis on a commercial salmon farm in British Columbia, Canada. The two replicate groups differed in early rearing conditions, time of introduction to sea water, and netpen environment. Family mortality data revealed a significant genetic component to disease resistance (sire and dam). Cumulative mortalities in the two replicate groups were significantly different, at 8.1% and 5.7%. There were no significant genotype-by-environment interactions, indicating that the genetic contribution to the disease resistance was stable across environments. Plasma lysozyme activity in both environmental groups was found to have a significant genetic component (sire). There was a significant positive correlation between plasma lysozyme activity and vibriosis mortality.     

Experimental evidence on prevention of infection by the ectoparasitic protozoans Ichthyobodo salmonis and Trichodina truttae in juvenile chum salmon using ultraviolet disinfection of rearing water

In northern Japan, juvenile chum salmon Oncorhynchus keta (Walbaum) are released from hatcheries to enhance the fishery resource. Infections with ectoparasitic protozoans, particularly the flagellate Ichthyobodo salmonis and the ciliate Trichodina truttae, occasionally cause severe mortality among hatchery‐reared juveniles. This study examined the susceptibility of the two parasites to wide‐ranging UV irradiation (experiment 1) and then investigated whether UV disinfection of the rearing water using a commercial device was useful for preventing infections among juveniles in a small‐scale rearing system over a 28‐day period (experiment 2). In experiment 1, parasite mortality reached 100% with UV irradiation doses of ≥9.60 × 10⁵ μW s/cm² for I. salmonis and ≥8.40 × 10⁵ μW s/cm² for T. truttae. In experiment 2, disinfection of the rearing water at a UV irradiation dose of 2.2 × 10⁶ μW s/cm² succeeded in complete prevention of both parasites in the juvenile salmon. These results elucidate the minimum dose of UV irradiation for inactivation of I. salmonis and T. truttae, and demonstrate the usefulness of water disinfection using a commercial UV irradiation device to prevent infections by these parasites in hatchery‐reared juvenile chum salmon.     

Juvenile Chinook salmon,Oncorhynchus tshawytscha,growth and diet in riverine habitat engineered to improve conditions for spawning

Abstract Many habitat enhancement techniques aimed at restoring salmonid populations have not been comprehensively assessed. The growth and diet of juvenile Chinook salmon, Oncorhynchus tshawytscha (Walbaum), rearing in a reach designed to enhance spawning were evaluated to determine how a non‐target life stage fared in the engineered habitat. Prior work demonstrated differences in food web structure between restored and unenhanced reaches of the Merced River, thus juvenile salmon feeding dynamics were also hypothesised to vary. Dependent variables were compared among fish collected from within and near the upper boundary of the restored reach and in an unenhanced habitat upstream. Diets, otolith‐derived growth and stable isotope‐inferred trophic positions were compared. Baetidae mayflies were particularly important prey in the restored reach, while elsewhere individuals exhibited heterogeneous diets. Salmon residing at the bottom of the restored reach exhibited slightly faster growth rates relative to fish collected elsewhere, although stable isotope and diet analyses suggested that they fed at a relatively low trophic position. Specialised Baetis predation and/or abundant interstitial refugia potentially improved rearing conditions in the restored reach. Data suggest that gravel enhancement and channel realignment designed to augment adult spawning habitat may simultaneously support juvenile Chinook salmon despite low invertebrate food resources.     

Laboratory and field investigations of salmon lice [Lepeophtheirus salmonis (Krøyer)] infestation on Atlantic salmon (Salmo salar L.) post-smolts

Hatchery‐reared 1‐year‐old Atlantic salmon post‐smolts (Salmo salar L.), artificially infected with salmon lice [Lepeophtheirus salmonis (Krøyer)] copepodids, were found to suffer from primary alterations (increased cortisol levels) at early lice stages. Secondary alterations, such as osmotic stress (increased chloride levels), first occurred after the preadult stages of the lice appeared. Fish with the highest salmon lice infections died throughout the experiment. Seven years of field investigation of Trondheimsfjorden showed that Atlantic salmon post‐smolts descending coastal waters can become heavily infected with salmon lice. The migrating post‐smolts were only infected with the chalimus stages, showing that the fish had only recently left the rivers. The infection level, however, varied considerably between the years, and, in 1998, the infection was higher than previous years. The experimental results have been combined with the field data to appraise the consequences of the infection.     

Large-scale, differential summer habitat use of three anadromous salmonids in a large river basin in Oregon, USA

The summer distribution of three juvenile anadromous salmonid species was compared in the mainstem sections and nine tributaries of the Upper South Umpqua River Basin, Oregon, USA. Chinook salmon, Oncorhynchus tshawytscha (Walbaum), was found at highest densities in the mainstem and mid-elevation tributaries. Coho salmon, O . kisutch (Walbaum), was essentially absent from the mainstem, but was found at high densities in the low-elevation tributaries. Steelhead trout, O . mykiss (Walbaum), was found in the mainstem and all nine tributaries; the highest densities of this species were in the upper mainstem reaches and high-elevation tributaries. No significant correlations were found between ranks of mainstem and tributary densities among the three species (Spearman's rs; P > 0.05). Observed spatial segregation of basin habitat by these species during summer indicated that protection provided at the level of the river basin will be necessary to assure the continued existence of all stocks.     

Evaluation of non‐traditional modelling techniques for forecasting salmon returns

Forecasting adult salmon abundance is problematic when the number of observations is small relative to the number of potential explanatory variables. Machine learning and other non‐traditional techniques employ algorithms designed to prevent model overfitting. Data from 18 coho salmon, Oncorhynchus kisutch (Walbaum), and seven Chinook salmon, Oncorhynchus tschawytscha (Walbaum), populations on the Oregon coast were used to evaluate the forecast performance of artificial neural networks, elastic net, least absolute shrinkage and selection operator, principal component regression (PCR) and ridge regression (RR) compared to several more traditional techniques. In general, the non‐traditional modelling techniques evaluated in this study performed similarly to the traditional techniques with the exception of sibling regression. This suggests that they have merit for improving actual predictions. Among the non‐traditional techniques, PCR resulted in the lowest prediction error for the coho salmon populations, and RR predicted Chinook salmon returns most accurately. The techniques explored are not an easy solution to a difficult problem. Spurious conclusions about the processes that generate salmon returns still may result as evidenced by the inclusion of an unrelated variable in many of the non‐traditional models.     

Carcass selenium loss as an indicator of stress in barge transported chinook salmon (Oncorhynchus tshawytscha Walbaum) smolts

Two salmon smolt barge transport experiments were conducted to measure tissue trace element selenium (Se) loss during the 30 h transport 500 km downriver past seven hydroelectric dams on the Columbia River. Carcass Se was measured before and after the barge trip. Liver glutathione peroxidase (GTPX) activity and total ascorbate concentrations were assayed to correlate Se loss with GTPX levels. Hatchery‐reared smolt chinook salmon, Oncorhynchus tshawytscha (Walbaum), were tested before and after barging. Liver ascorbate concentrations, measured by high‐performance liquid chromatography techniques, showed levels adequate to support GTPX activity. Salmon smolts lost up to 20% of carcass tissue Se during the 30 h barge confinement. Selenium was analysed by analytical polarography. Samples collected after two barging episodes showed Se loss and concomitant elevated liver GTPX activity after the transport. Tissue Se loss may be useful to measure stress in fish, and increasing tissue Se in hatchery salmon smolts prior to release and transport may be warranted.     

Microhabitat selection of Gyrodactylus salaris Malmberg on different salmonids

The microhabitat selection of the ectoparasite Gyrodactylus salaris (Lærdalselva strain, Norway) was investigated concurrently with studies on the parasite population growth on five strains of Atlantic salmon, Salmo salar L., and a strain of Danish rainbow trout, Oncorhynchus mykiss (Walbaum). The salmon used were hatchery‐reared parr of East Atlantic strains [River Conon (Scotland), River Storå (western Denmark) and River Ätran (western Sweden)] and Baltic strains [Lule and Ume (eastern Sweden)]. The location and numbers of parasites were recorded on anaesthetized fish once a week from week 0 to week 8. The mean abundance of G. salaris steadily increased to high levels on the River Conon, Storå and Ätran strains until the end of the experiment. The mean abundance of G. salaris on the two Baltic strains (River Lule älv and River Ume älv) initially increased but after 4–7 weeks the growth of the parasite infrapopulations decreased markedly. The Danish rainbow trout strain showed the lowest abundances of all the fish species and strains. Gyrodactylus salaris preferentially selected the fins and head region when colonising the hosts (all species and strains). Increasing percentages of G. salaris on the tail fins of the East Atlantic strains and rainbow trout were found during the course of infection, whereas the two Baltic salmon strains experienced a decreasing percentage of parasites in this microhabitat.     

Heavy Metal Content of Coho Oncorhynchus kisutch and Chinook Salmon O. tschawytscha Reared in Deep Upwelled Ocean Waters in Hawaii1

Coho (Oncorhynchus kisutch) and chinook salmon (O. tschawyrscha) were reared in seawater pumped from the 600 m depth at the Natural Energy Laboratory of Hawaii. Fish were reared in either 100% deep water at 11 C, or a mixture of 55% deep and 45% surface waters at 17 C. Salmon flesh was analyzed for mercury, zinc, cadmium and lead. Mercury concentration in coho salmon reared at 11 C was significantly lower than the concentration in coho reared at 17 C (P < 0.05), but other heavy metal concentrations in tissue did not differ between the two species or between the two rearing temperatures. Compared with wild salmon from the northwest Pacific Ocean, salmon reared in Hawaii had significantly lower concentrations of mercury and lead (P < 0.05), but significantly higher concentrations of zinc and cadmium. These differences in heavy metal concentration between wild and cultured salmon were most readily explained by differences in the heavy metal content of diets and the lack of heavy metal concentrations in rearing water. The concentration of mercury in salmon tissue was at least 20 times less than the 1.0 μ/g U.S. Food and Drug Administration action level for mercury. Action levels have not been established for zinc, cadmium, and lead.