Gill pathology in Scottish farmed Atlantic salmon,Salmo salar L., associated with the microsporidian Desmozoon lepeophtherii Freeman et Sommerville, 2009

Gill disorders have emerged in recent years as a significant problem in the production of marine‐stage Atlantic salmon Salmo salar L. The multi‐aetiological condition ‘proliferative gill inflammation’ (PGI) has been reported to cause heavy losses in western Norway, yet reports of Scottish cases of the disease have remained anecdotal. In the present study, histopathological material from a marine production site in the Scottish Highlands experiencing mortalities due to a seasonal gill disease with proliferative‐type pathology was examined using light microscopy, special staining techniques and transmission electron microscopy (TEM). The microsporidian Desmozoon lepeophtherii Freeman et Sommerville, 2009 (syn. Paranucleospora theridion) was identified by staining using a Gram Twort method and TEM associated with distinctive proliferative and necrotic pathology confined to the interlamellar Malpighian cell areas of the primary filaments. Epitheliocystis was not a feature of the gill pathology observed. It is believed this is the first report of D. lepeophtherii being identified associated with pathology in a Scottish gill disease case, and supports anecdotal reports that a disease at least partly synonymous with PGI as described by Norwegian researchers is present in Scottish aquaculture.     

Parasite fecundity decreases with increasing parasite load in the salmon louse Lepeophtheirus salmonis infecting Atlantic salmon Salmo salar

Aggregation is common amongst parasites, where a small number of hosts carry a large proportion of parasites. This could result in density‐dependent effects on parasite fitness. In a laboratory study, we explored whether parasite load affected parasite fecundity and survival, using ectoparasitic salmon lice (Lepeophtheirus salmonis Krøyer, 1837) infecting Atlantic salmon (Salmo salar) hosts. We found a significant reduction in fecundity with higher parasite load, but no significant effect on survival. Together with previous findings, this suggests that stronger competition amongst female lice under high parasite load is a more likely explanation than increased host immune response.     

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.     

Characterization of Mycobacterium salmoniphilum as causal agent of mycobacteriosis in Atlantic salmon,Salmo salar L., from a freshwater recirculation system

Thirty Atlantic salmon, Salmo salar L., with low corporal condition relative to other fish present in the culture system, were sampled from a freshwater recirculation pisciculture located in Chile. The most characteristic signs and lesions were cachexia and presence of multiple greyish‐white granulomas within internal organs. The external and internal lesions, along with the microscopic, histologic and biochemical findings, were consistent with mycobacteriosis. The identification of Mycobacterium salmoniphilum as the causal agent of the lesions was possible through the use of molecular analyses. This study represents the first report of Mycobacterium salmoniphilum in a freshwater recirculation system and the first case of fish mycobacteriosis described in Chile.     

Preliminary success using hydrogen peroxide to treat Atlantic salmon,Salmo salar L., affected with experimentally induced amoebic gill disease (AGD)

Currently, the only effective and commercially used treatment for amoebic gill disease (AGD) in farmed Tasmanian Atlantic salmon is freshwater bathing. Hydrogen peroxide (H₂O₂), commonly used throughout the aquaculture industry for a range of topical skin and gill infections, was trialled in vitro and in vivo to ascertain its potential as an alternative treatment against AGD. Under in vitro conditions, trophozoites of Neoparamoeba perurans were exposed to three concentrations of H₂O₂ in sea water (500, 1000 and 1500 mg L^?1) over four durations (10, 20, 30 and 60 min) each at two temperatures (12 and 18 °C). Trophozoite viability was assessed immediately post‐exposure and after 24 h. A concentration/duration combination of 1000 mg L^?1 for >10 min demonstrated potent amoebicidal activity. Subsequently, Atlantic salmon mildly affected with experimentally induced AGD were treated with H₂O₂ at 12 and 18 °C for 15 min at 1250 mg L^?1 and their re‐infection rate was compared to freshwater‐treated fish over 21 days. Significant differences in the percentage of filaments affected with hyperplastic lesions (in association with amoebae) and plasma osmolality were noted between treatment groups immediately post‐bath. However, the results were largely equivocal in terms of disease resolution over a 3‐week period following treatment. These data suggest that H₂O₂ treatment in sea water successfully ameliorated a clinically light case of AGD under laboratory conditions.     

The effect of dietary chitin on growth and nutrient digestibility in farmed Atlantic cod,Atlantic salmon and Atlantic halibut

The effect of adding 0%, 1%, 2% and 5% chitin from prawn shells in the diets for Atlantic cod, Atlantic halibut and Atlantic salmon on growth was investigated. Nutrient digestibility and feed utilization was investigated in salmon and cod. Atlantic cod grew from 186 ± 29 to 383 ± 78 g (N = 960) over 13 weeks. Dietary chitin had no effect on length, weight, condition, liver size or specific growth rate (SGR). The apparent digestibility (ADC) for protein ranged from 84.7% to 86.5%, lipid between 88.8% and 93.1% and dry matter from 96.1% to 96.6%. Feed utilization varied between 1.08 and 1.11 and was not correlated with dietary chitin content. Atlantic salmon tripled their weight from 199 ± 9 to 615 ± 75 g (N = 480) during the 13 weeks. High inclusions of chitin (>1%) reduced both growth rate and condition. Protein and lipid ADC was negatively correlated with dietary chitin. Feed utilization ranged between 0.86 and 0.90 and was not significantly affected by dietary chitin. Faecal protein increased significantly with increasing dietary chitin, while faecal dry matter and lipid did not. Individually tagged Atlantic halibut grew from 1300 ± 470 to 2061 ± 714 g (N = 70) during 6 months. Individual growth rates varied within each group from being slightly negative to 0.81%·day^?1. Diet had no significant effect on growth rates. Atlantic cod and Atlantic halibut seems unaffected by up to 5% chitin additions in the diet, while chitin >1% of diet negatively affects growth and nutrient utilization in Atlantic salmon.     

Salmon gill poxvirus,a recently characterized infectious agent of multifactorial gill disease in freshwater‐ and seawater‐reared Atlantic salmon

Gill diseases cause considerable losses in Norwegian salmon farming. In 2015, we characterized salmon gill poxvirus (SGPV) and associated gill disease. Using newly developed diagnostic tools, we show here that SGPV infection is more widely distributed than previously assumed. We present seven cases of complex gill disease in Atlantic salmon farmed in seawater and freshwater from different parts of Norway. Apoptosis, the hallmark of acute SGPV infection, was not easily observed in these cases, and qPCR analysis was critical for identification of the presence of SGPV. Several other agents including Costia‐like parasites, gill amoebas, Saprolegnia spp. and bacteria were observed. The studied populations experienced significant mortalities, which increased to extreme levels when severe SGPV infections coincided with smoltification. SGPV infection appears to affect the smoltification process directly by affecting the gills and chloride cells in particular. SGPV may be considered a primary pathogen as it was often found prior to identification of complex gill disease. It is hypothesized that SGPV‐induced gill damage may impair innate immunity and allow invasion of secondary invaders. The distinct possibility that SGPV has been widely overlooked as a primary pathogen calls for extended use of SGPV qPCR in Atlantic salmon gill health management.     

How are the salmon lice (Lepeophtheirus salmonis Krøyer, 1837) in Atlantic salmon farming affected by different control efforts: A case study of an intensive production area with coordinated production cycles and changing delousing practices in 2013–2018

The aims of the present study were to describe the salmon lice (Lepeophtheirus salmonis Krøyer, 1837) situation in an intensive salmon production area in mid‐Norway and to consider implications of changing practices of how salmon lice infestation can be controlled. The results in this study suggest that there are steps that can be carried out to keep salmon lice under control even during years when the temperature facilitates a quick salmon lice development. The present work indicates that the use of cleaner fish can delay the time it takes adult female lice to reach 0.1 per salmon in the beginning of a production cycle. It suggests that the timing of cleaner fish deployment into salmon cages can influence its effectiveness in controlling salmon lice. It also gives caution to letting salmon lice develop unchecked, even at levels far below the current lice limit, because of the difficulties to control salmon lice when the external infection pressure is too high. This study took place during a rapid change in delousing methods, in an area with coordinated salmon production. Despite its exploratory nature, this study offers insights into the salmon lice fluctuations in relation to efforts aimed at controlling it.     

Effects of feeding rate and frequency on growth performance,digestion and nutrients balances of Atlantic salmon (Salmo salar) in recirculating aquaculture systems (RAS)

A 10‐week study was conducted to investigate the effects of feeding rate and frequency on growth performance, digestion and nutrients balances of Atlantic salmon (Salmo salar) in replicated recirculating aquaculture systems (RAS). Replicated groups of juvenile salmon weighing 90 ± 2.5 g (mean ± SD) were fed a commercial feed (21.63 MJ kg^?1 gross energy) to designed feeding rate (1.4%, 1.6% and 1.8% body weight day^?1, BW day^?1) and feeding frequency (2 and 4 meals day^?1) combinations. Specific growth ratio varied between 1.15 ± 0.02 and 1.37 ± 0.16% day^?1, and feed conversion ratio ranged from 0.96 ± 0.03 to 1.16 ± 0.02. The nitrogen and phosphorus retention rates were from 36.50 ± 1.94 to 47.08 ± 5.23% and from 20.42 ± 1.05 to 38.59 ± 2.80%. Apparent digestibility coefficients (ADC) in dry matter, protein, lipid and energy showed no significant differences for all groups. However, fish fed at 1.6% BW day^?1 and 4 meal day^?1 groups had relatively better growth and nutrient retention efficiency compared to other groups. In addition, concentrations of nitrogenous and phosphorous compounds were also detected in this study. These results suggested that salmon of 100–200 g in RAS could in practice be fed at 1.6% BW day^?1 and 4 meals day^?1, taking environmental impacts into account.     

Reduced total hardness of fresh water enhances the efficacy of bathing as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L

The current treatment for amoebic gill disease (AGD)-affected Atlantic salmon involves bathing sea-caged fish in fresh water, often sourced from local dams, for 3-4 h. In both a small-scale laboratory and an on-farm field experiment, the effects of water hardness on the efficacy of freshwater bathing were assessed. Results showed that soft fresh water (19.3-37.4 mg L(-1) CaCO3), whether it be naturally soft city mains water or artificially softened dam water, was more efficacious at alleviating AGD in affected fish than hard fresh water (173-236.3 mg L(-1) CaCO3). Soft freshwater bathing significantly reduced viable gill amoebae numbers (from 73.9 to 40.9% of total count) and significantly alleviated gill pathology, both gross and histological. Following bathing, gross gill pathological scores of soft freshwater bathed fish lagged 2 weeks behind hard freshwater bathed fish. Significant gill lesion fragmentation, and shedding of lesion-associated hyperplastic tissue, was accompanied by a significant reduction in AGD-affected gill filaments in soft freshwater bathed fish. Furthermore, soft freshwater bathing alleviated the blood plasma electrolyte imbalance seen in control (sea water) and hard freshwater bathed fish. This study showed that the use of soft fresh water for bathing AGD-affected Atlantic salmon could be an improvement to the current method of treatment. Not only does it reduce gill amoeba numbers, but also, it is of a therapeutic advantage with the potential to reduce bathing frequency.     

Performance and welfare of Atlantic salmon,Salmo salar L. post-smolts in recirculating aquaculture systems: Importance of salinity and water velocity

Producing a larger post-smolt in recirculating aquaculture systems (RAS) could shorten the production time in sea cages and potentially reduce mortality. Knowledge of the biological requirements of post-smolts in closed-containment systems is however lacking. In the present study, the effects of salinity and water velocity on growth, survival, health, and welfare of Atlantic salmon reared in RAS were examined. Salmon smolts were stocked in three separate RAS with salinities of 12, 22, and 32‰ and subjected to high (1.0 body lengths per s⁻¹) or low (0.3 body lengths second⁻¹) water velocity. Growth performance, survival, welfare, and physiological stress responses were monitored until the fish reached a bodyweight of around 450 g. Growth rate was higher at lower salinity and higher water velocity generally had a positive effect on growth in all salinities. Feed conversion ratio was lower at 12‰ compared to the 22 and 32‰ when the fish were between 250 and 450 g. Higher mortality, elevated plasma cortisol levels, higher incidence of cataract, and a higher expression of stress-induced genes in the skin (iNOS, Muc5ac-like) indicated a negative effect of higher salinity on fish welfare. Male maturation was low (<1%), and not affected by salinity or water velocity.     

The mutagenesis of a type IV secretion system locus of Piscirickettsia salmonis leads to the attenuation of the pathogen in Atlantic salmon,Salmo salar

Piscirickettsiosis is a threatening infectious disease for the salmon industry, due to it being responsible for significant economic losses. The control of outbreaks also poses considerable environmental challenges. Despite Piscirickettsia salmonis having been discovered as the aetiological agent of the disease more than 25 years ago, its pathogenicity remains poorly understood. Among virulence factors identified so far, type four secretion systems (T4SS) seem to play a key role during the infection caused by the bacterium. We report here the genetic manipulation of P. salmonis by means of the transference of plasmid DNA in mating assays. An insertion cassette was engineered for targeting the icmB gene, which encodes a putative T4SS‐ATPase and is carried by one of the chromosomal T4SS clusters found within the genome of P. salmonis PM15972A1, a virulent representative of the EM‐90‐like strain. The molecular characterization of the resulting mutant strain demonstrated that the insertion interrupted the target gene. Further in vitro testing of the icmB mutant showed a dramatic drop in infectivity as tested in CHSE‐214 cells, which is in agreement with its attenuated behaviour observed in vivo. Altogether, our results demonstrate that, similar to other facultative intracellular pathogens, P. salmonis’ virulence relies on an intact T4SS.     

脂肪和蛋白质水平对工业养殖大西洋鲑消化酶、非特异性免疫及水质的影响

为探讨脂肪和蛋白质水平对工业化养殖大西洋鲑相关消化酶活力、免疫及对水质指标的影响,采用3×2双因素试验设计(3脂肪水平:18%、21%、24%,即F18、F21、F24;2蛋白质水平:38%、48%,即P38、P48),形成6种实验处理的膨化配合颗粒料,每处理3重复,在工业化封闭循环海水养殖(RAS)条件下,选用初重(650.0±45.50)g大西洋鲑720尾进行56 d的养殖实验。结果显示:(1)F21可显著提高大西洋鲑胃、肠、肝脂肪酶活力,比低脂肪分别提高11.52%、14.63%、4.31%;P48使肠、肝胰蛋白酶活力分别提高8.23%、8.33%,且发现其活力水平远高于胃蛋白酶。(2)F18、P48可显著提高肠道AKP酶活力,F18比F21、F24分别提高18.61%、31.70%,P48比P38提高13.69%。(3)F21、F24可显著改善血清的抗氧化能力,F21比F18的SOD活力提高10.32%,同时MDA含量降低4.49%;P48有利于提高血清LZM活力及补体C3含量,P48比P38分别高9.49%、5.93%。(4)低蛋白质水平可显著降低水体中氨氮、硝酸盐的含量,P38比P48分别降低61.70%、28.36%;提高脂肪水平可降低水体氨氮含量,F21、F24比F18分别降低10.00%、8.20%。研究表明,饲料脂肪和蛋白质水平与消化吸收酶、非特异免疫、养殖水氨氮之间关系特征明显;适当提高脂肪水平有利于提高消化道脂肪酶活力,提高蛋白质水平有利于提高免疫力;低蛋白质和中高脂肪组合可有效降低大西洋鲑的氨氮排泄量。     

Recovery of Bacillus mycoides,B. pseudomycoides and Aeromonas hydrophila from common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) with gill disease

During a 3‐month period from June to the end of August 2016, ~5% mortalities were observed in a farm with rainbow trout (Oncorhynchus mykiss Walbaum) and one farm of common carp (Cyprinus carpio L.) in Bulgaria. The disease was manifested by gill ulcers/rot, asphyxiation and bloody ascites. Aeromonas hydrophila was isolated from the internal organs of all the diseased fish. Bacillus mycoides or B. pseudomycoides were recovered from the gill lesions on diseased carp and rainbow trout, respectively, with identification achieved by conventional phenotyping and by sequencing of the 16S rRNA gene. In vivo experiments confirmed that all three organisms were pathogenic to rainbow trout.     

Colonization of Aeromonas salmonicida subsp. masoucida strains in Atlantic salmon (Salmo salar L.) during infection

Aeromonas salmonicida subsp. masoucida (ASM) is classified as atypical A. salmonicida and brought huge economic damages to the local salmonid aquaculture in China. An ASM strain named AS‐C4 was used to investigate the colonization of ASM in Atlantic salmon (Salmo salar L.) by an immersion challenge with the control group (T0, no AS‐C4), group T1 (2.67 × 10⁴ CFU/ml AS‐C4) and group T2 (2.67 × 10⁷ CFU/ml AS‐C4). The numbers of AS‐C4 copies in different fish tissues (gill, intestine, skin, blood, muscle, spleen, liver and kidney) were determined at different time points post challenge using the quantitative real‐time PCR (qRT‐PCR). AS‐C4 were detected in the gill and intestine as early as 0 hr after the challenge both in T1 and T2 groups, suggesting that the gill and intestine were probably the portals of entry of AS‐C4 into salmon. Although AS‐C4 could not be detected in the skin until 24 hr after the challenge in T1 group, it could be detected in the skin as early as 0 hr after the challenge in T2 group, indicating that the skin may also be a portal of entry of AS‐C4 into salmon. AS‐C4 was immediately detected in the blood within 3 hr after it entered the host, suggesting that AS‐C4 successfully invaded the bloodstream of fish. After AS‐C4 colonized the host, it colonized the internal tissues, such as the spleen, liver, kidney and muscle. The results of this study will contribute to the understanding of the pathogenesis of the ASM strains and give a broader understanding of the infection route of ASM in it's host, providing more information for the development of new therapeutic strategies to protect against this pathogen in aquaculture.     

Use of a growth model to assess the suboptimal growth of Atlantic salmon parr in a subarctic river

This study tested the hypothesis that Atlantic salmon parr grew suboptimally according to the water temperature regime during the ice‐free season (from May to September) in the subarctic River Reisa (northern Norway). A laboratory‐based growth model was used to estimate the optimal growth which was then compared with the observed growth of the parr in the river. Water temperatures were recorded every second hour with a submersible data logger, and observed and predicted growth of several age classes were compared at two nearby nursery habitats. Except for parr aged 2, most age classes showed no significant change, or even a weight loss, from May to July, whereas from July to September, positive growth was found for all age classes coinciding with favourable temperatures above a critical temperature for growth. The parr growth rates were suboptimal, being lower than the predicted values, but values for observed and predicted body mass were not significantly different in five of 12 comparisons. Hence, the fish were sometimes unable to grow according to the optimal baseline set by the temperature regime. Therefore, factors other than temperature, such as limited food resources and resource competition, may also be a major determinant of Atlantic salmon parr growth. Our study demonstrates how growth predictions based on mathematical models may be employed by fish ecologists to identify growth deficiencies in wild fish populations.