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A better understanding of unique anatomical and functional features of the visual systems of teleost fish could provide key knowledge on how these systems influence the health and survival of these animals in both wild and culture environments. We took a systematic approach to assess some of the visual systems of spotted wolffish (Anarhichas minor), a species of increasing importance in North Atlantic aquaculture initiatives. The lumpfish (Cyclopterus lumpus) was included in these studies in a comparative manner to provide reference. Histology, light and electron microscopy were used to study the spatial distribution and occurrence of cone photoreceptor cells and the nature of the retinal tissues, while immunohistochemistry was used to explore the expression patterns of two photoreceptor markers, XAP‐1 and XAP‐2, in both species. A marine bacterial infection paradigm in lumpfish was used to assess how host–pathogen responses might impact the expression of these photoreceptor markers in these animals. We define a basic photoreceptor mosaic and present an ultrastructural to macroscopic geographical configuration of the retinal pigment tissues in both animals. Photoreceptor markers XAP‐1 and XAP‐2 have novel distribution patterns in spotted wolffish and lumpfish retinas, and exogenous pathogenic influences can affect the normal expression pattern of XAP‐1 in lumpfish. Live tank‐side ophthalmoscopy and spectral domain optical coherence tomography (SD‐OCT) revealed that normal cultured spotted wolffish display novel variations in the shape of the retinal tissue. These two complementary imaging findings suggest that spotted wolffish harbour unique ocular features not yet described in marine teleosts and that visual function might involve specific retinal tissue shape dynamics in these animals. Finally, extensive endogenous biofluorescence is present in the retinal tissues of both animals, which raises questions about how these animals might use retinal tissue in novel ways for visual perception and/or communication. This work advances fundamental knowledge on the visual systems of two economically important but now threatened North Atlantic teleosts and provides a basic foundation for further research on the visual systems of these animals in health versus disease settings. This work could also be useful for understanding and optimizing the health and welfare of lumpfish and spotted wolffish in aquaculture towards a one health or integrative perspective. [ABSTRACT FROM AUTHOR]

Both in their natural habitat (estuaries, marine coastal environment, ocean depths and freshwater lakes) or in captivity conditions (land‐based, open‐sea aquaculture and aquariums), exposure to hypoxia can have direct detrimental effects on fish growth, reproduction, behaviour and health. The objective of this study was to investigate the effects of chronic exposure of spotted wolffish (Anarhichas minor) (∼1.4 kg) and the interspecific hybrid (A. minor × A. lupus) (∼600 g) to moderate hypoxia levels (dissolved oxygen [DO]: 40%, 50% and 60% saturation) in comparison to a normoxia group (control 100%) over a period of 14 weeks at ∼7.5°C. The trials were conducted as a common‐garden experiment (both fish groups reared together) in quadruplicate (4 tanks per DO level = 16 tanks). Fish performance (specific growth rate, feed intake [FI]) and physiological status (condition factor [K] and hepatosomatic index, haematocrit) were monitored at different intervals. The metabolic enzyme activity of citrate synthase (CS), lactate dehydrogenase (LDH) and pyruvate kinase (PK) was measured as well as antioxidant enzymes glutathione reductase and catalase. Oxidative stress indices were assessed by the quantification of thiobarbituric acid reactive substances (TBARS) and the measurement of the enzymatic activity level of aconitase. Significant reduction in growth rate occurred at DO 40%. DO was identified as a major driver of FI. Our results indicate a very strong adaptation to hypoxia as (1) exposures to DO 40% were not severe enough to affect aerobic or anaerobic metabolism capacity or to significantly induce oxidative stress and (2) metabolic organization of the muscle tissue, as expressed by LDH, CS and PK is not affected by DO levels down to 40%. However, at the lowest DO levels (40% and 50%), hybrid fish displayed significantly better growth than the spotted wolffish. [ABSTRACT FROM AUTHOR]

Abstract Both in their natural habitat (estuaries, marine coastal environment, ocean depths and freshwater lakes) or in captivity conditions (land‐based, open‐sea aquaculture and aquariums), exposure to hypoxia can have direct detrimental effects on fish growth, reproduction, behaviour and health. The objective of this study was to investigate the effects of chronic exposure of spotted wolffish (Anarhichas minor) (∼1.4 kg) and the interspecific hybrid (A. minor × A. lupus) (∼600 g) to moderate hypoxia levels (dissolved oxygen [DO]: 40%, 50% and 60% saturation) in comparison to a normoxia group (control 100%) over a period of 14 weeks at ∼7.5°C. The trials were conducted as a common‐garden experiment (both fish groups reared together) in quadruplicate (4 tanks per DO level = 16 tanks). Fish performance (specific growth rate, feed intake [FI]) and physiological status (condition factor [K] and hepatosomatic index, haematocrit) were monitored at different intervals. The metabolic enzyme activity of citrate synthase (CS), lactate dehydrogenase (LDH) and pyruvate kinase (PK) was measured as well as antioxidant enzymes glutathione reductase and catalase. Oxidative stress indices were assessed by the quantification of thiobarbituric acid reactive substances (TBARS) and the measurement of the enzymatic activity level of aconitase. Significant reduction in growth rate occurred at DO 40%. DO was identified as a major driver of FI. Our results indicate a very strong adaptation to hypoxia as (1) exposures to DO 40% were not severe enough to affect aerobic or anaerobic metabolism capacity or to significantly induce oxidative stress and (2) metabolic organization of the muscle tissue, as expressed by LDH, CS and PK is not affected by DO levels down to 40%. However, at the lowest DO levels (40% and 50%), hybrid fish displayed significantly better growth than the spotted wolffish.

In spotted wolffish Anarhichas minor aquaculture, cryopreservation is used to secure sperm availability throughout the entire spawning season. Under current protocols, sperm is cryopreserved in 0.5-mL straws. This implies thawing a considerable number of straws for insemination with cryopreserved sperm. In this work, we scale up the spotted wolffish sperm cryopreservation procedure through the development of a protocol for sperm cryopreservation in 5-mL cryovials. Different freezing (distances from the liquid nitrogen surface) and thawing rates were tested. The best results were obtained with cryovials frozen at a distance of 1.5 cm from the liquid nitrogen surface and thawed either at 15 or 10 °C for 4 and 6 min, respectively. Under these conditions, similar percentage of motile cells, sperm velocity and percentage of viable cells were obtained in comparison with the sperm cryopreserved in the traditional 0.5-mL straws. This protocol will facilitate the process of insemination with cryopreserved sperm in the spotted wolffish hatcheries. [ABSTRACT FROM AUTHOR]

Spotted wolffish Anarhichas minor reproduction in captivity is dependent on in vitro fertilization. However, it is often challenging to acquire sufficient fresh sperm to fertilize the eggs that are obtained. In this study, we evaluate the possibility to store spotted wolffish sperm by refrigeration. Spotted wolffish sperm has the particularity that is already motile on stripping, and currently it is not possible to immobilize and reactivate. Thus, sperm refrigeration protocols should focus in extending this motility period that usually lasts up to 2 days. In a first experiment, we evaluated the possibility that the motility period of the sperm was limited by contamination with urine. The urea concentration in the sperm obtained both by stripping (17.10 ± 1.98 mg/dL) and directly from the testis (12.59 ± 2.37 mg/dL) was similar (p > 0.05), which indicate that the sperm collection method used avoid contamination with urine. Afterwards, we tested the possibility that the sperm motility period was limited by energy stores. The ATP concentration (initial value 5.65 ± 0.86 nmol/109 cells) remained stable (p = 0.099) during 30 h after sperm collection, and similar values (p = 0.329) were recorded at end of sperm storage in both diluted (3.88 ± 1.35 nmol/109 cells) and undiluted samples (4.76 ± 1.08 nmol/109). This indicates that the low intracellular ATP consumption, derived from the slow sperm motility, can probably be compensated rapidly enough by mitochondrial synthesis of ATP in the spotted wolffish sperm. In both experiments, diluted sperm kept higher percentage of motile cells during the storage time. [ABSTRACT FROM AUTHOR]

The first artificially fertilized spotted wolffish (Anarhichas minor) eggs hatched in Norway in the mid-1990s as this species was considered by Norwegian authorities to be a top candidate species for cold-water aquaculture in the North Atlantic regions. Previous research conducted in Norway (since 1992) and Canada (since 2000), focused on identifying key biological parameters for spotted wolffish cultivation which led, respectively, to the rapid establishment of a full commercial production line in northern Norway, while Québec (Canada) is witnessing its first privately driven initiative to establish commercial production of spotted wolffish on its territory. The control of reproduction can be viewed as a major requirement to achieve the development of performant strains using genetic selection tools and/or all-year-round production to bring about maximal productivity and synchronization among a given captive population. Although the basic reproduction aspects are more understood and controlled there are still some challenges remaining involving broodstock and upscaling of operations that limit the achievement of a standardized production at the commercial level. Quality of gametes is still considered a major constraint and it can be affected by multiple factors including nutrition, environmental conditions, handling practices, and welfare status. Internal insemination/fertilization and the protracted incubation period are challenging as well as the establishment of a health monitoring program to secure large-scale operations. The profound progress achieved in the control of reproduction, sperm handling, and cryopreservation methods for this species is presented and discussed. In this review, we also go into detail over the full range of up-to-date cultivation practices involving broodstock and identify areas that could benefit from additional research efforts (i.e., broodstock nutrition, health and welfare, scaling-up egg and larval production, genetics, and development of selective breeding programs).

Wolffish are regularly housed in aquaria, but little data on their husbandry and health is available for caretakers. High occurrence rates of nephrocalcinosis and urolithiasis have been observed in Atlantic Anarhichas lupus and spotted A. minor wolffish housed at 2 Canadian zoological institutions. To explore the effect of diet on nephrocalcinosis and urolithiasis development, a 16 mo prospective study was conducted. A total of 32 juvenile spotted wolffish were randomly assigned to one of 4 experimental groups fed exclusively with the following diet: (1) Skretting® Europa 18 pellets; (2) Mazuri® LS Aquatic Carni-Blend Diet Formula; (3) vitamin-supplemented fish-based diet, and (4) vitamin-supplemented invertebrate-based diet. Urinalysis, radiographs, and complete necropsies were performed at the end of the study. None of the wolffish developed uroliths during the study period. All specimens fed with the fish-based and invertebrate-based diets developed nephrocalcinosis, whereas this condition was seen in 12.5 and 0% of the fish in the Skretting® and Mazuri® groups, respectively. Affected wolffish often presented with oxalate crystalluria and increased radiodensity of the posterior kidneys. Urinalysis and radiographic study were considered useful in the antemortem diagnosis of nephrocalcinosis. None of the previously published risk factors for the development of nephrocalcinosis in fish were supported by the results of this study. However, nutritional analyses of the 4 diets suggest that high dietary levels of gelatin or vitamin C or low levels of vitamin E could be potential risk factors for the development of nephrocalcinosis in spotted wolffish and thus warrant further study.

This study provides new and practical information to implement the use of photoperiodic manipulation for the control of the sexual maturation of Canadian 6+ Atlantic wolffish (Anarhichas lupus) and 3+ first-time spawners spotted wolffish (Anarhichas minor) from Canadian and Norwegian populations. Wolffish reproductive cycle (gametogenesis and oocyte maturation and spermiation) was monitored for 23 consecutive months. Control groups were held under a 12-month simulated natural photoperiod and treatment groups under an 8-month compressed photoperiod. Plasma sex steroid concentrations (estradiol-17β and 11-ketotestosterone), oocyte diameter growth, and milt production were assessed monthly. For all groups under study, fish subjected to the compressed photoperiod spawned 2-6 months earlier than the controls. Complete out-of-shift cycle was not achieved based on the completion of two reproduction cycles, and this is briefly discussed. Photoperiod treatment induced temporal shifts in sex steroid profiles, which are the likely mediators of altered timing of ovulation/final maturation. Photoperiod has a strong influence on the timing of wolffish maturation and could be used as an efficient and inexpensive tool to secure wolffish reproduction operations (year-round supply of egg and milt and/or timing with optimal temperature regimes). [ABSTRACT FROM AUTHOR]

Oocytes were sampled from Atlantic wolffish (Anarhichas lupus) and spotted wolffish (Anarhichas minor) to investigate the effect of fixation and storage in formalin on oocyte size and weight over time. The effect of formalin on oocyte size was dependent on the original size of the oocyte. As the effect of fixation was similar for the two species, a common formula for the conversion of fixed oocyte size/weight to fresh oocyte size/weight was developed. The relationships were negative and nonlinear, the smallest oocytes increased ∼11 % in diameter and ∼15 % in weight when fixed in formalin, whereas the largest oocytes decreased about 4 % in diameter and 7 % in weight. After fixation, oocyte diameter was stable for up to 2, but not 10 years and for up to 10 years for oocytes at CA and VIT stage, respectively. • Oocytes of Atlantic and spotted wolffish were sampled at two ovary development stages to examine the influence of formlin on oocyte size and weight. • The influence of formlin was dependent on the original size of the oocyte. • After fixation in formalin, oocyte size was stable for at least two years. [ABSTRACT FROM AUTHOR]

Vitellogenin (VTG) from spotted wolffish, Anarhichas minor, a candidate species for cold-water marine aquaculture, was purified by MgCl/EDTA precipitation followed by a two-step chromatographic procedure. VTG had an apparent molecular mass of 470 kDa, as determined by gel filtration, and an amino acid composition similar to those of other teleosts. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the purified VTG revealed a major band with a relative molecular weight of 166 kDa and some minor bands. Spotted wolffish VTG (sw-VTG) is relatively robust to in vitro degradation, as shown when samples of purified VTG and plasma from mature females subjected to various storage conditions or multiple freeze/thaw cycles were analyzed by Western blot. We developed an indirect competitive enzyme-linked immunosorbent assay (ELISA) using an antibody against Atlantic wolffish ( Anarhichas lupus) VTG and purified sw-VTG. The ELISA had a detection limit of 6.7 ng/ml and a working range of 16.2-787.5 ng/ml, with intra- and inter-assay coefficients of variation ranging from 1.5 to 7.3 % and 7.1 to 14.3 %, respectively. The assay could distinguish males from immature females and discriminate maturing females at different stage of oocyte development. These results suggest that the sw-VTG ELISA would be useful in spotted wolffish aquaculture to determine sex and monitor female maturation. [ABSTRACT FROM AUTHOR]

The purpose of this study was to determine the cortisol response of spotted wolffish (Anarhichas minor) after exposure to air immersion and netting (acute stress) under a range of increasing densities (chronic stressor). In addition, the presence of a cumulative negative impact from chronic stress was assessed by the application of an additional acute stressor following rearing at various densities, and proxies of behavior were evaluated (feeding and aggression data). The stress response to air immersion and netting led to increasing plasma cortisol values at 0.5, 13, and 37 h post-stress, with the highest value achieved after 13 h. At 168 h, cortisol concentrations returned to similar pre-stress value. Changes in cortisol concentrations were relatively low after acute stress (five to six fold increases) and recovery time long lasting (>37 h). Prolonged rearing at 20, 30, and 40 kg m−2showed a non-significant trend of increasing cortisol values with increasing density, and the cortisol response after the application of an additional acute stressor was significantly altered, i.e., higher cortisol levels were measured in unstressed (chronic stress only) compared to stressed fish (chronic stress + acute stress). Our results suggest that (1) spotted wolffish is a low responder that additionally displays a long-lasting cortisol elevation following an acute stressor; (2) fixed densities did not lead to significant differences of plasma cortisol concentration or aggression levels, although involved marked changes in daily feed intake and food conversion efficiency were observed; and (3) cortisol levels reached acute stress values after prolonged chronic stress. Although this species displays indications of being relatively resistant to handling disturbances during aquaculture practices, it is nevertheless responsive to chronic stress factors that could alter the physiological response to subsequent acute stressor. [ABSTRACT FROM AUTHOR]

Abstract: Growth rate is dependent upon adequate provision of amino acids especially in newly-hatched fish which experience very high growth rate. The replacement of a fraction of protein content by partially hydrolyzed (pre-digested) proteins was carried out and the digestive capacities and performances of larval/juvenile spotted wolffish (Anarhichas minor) were measured. The goal of this study was to verify whether the scope for growth is principally dictated by the proteolytic capacity of the digestive system by examining the effect of protein hydrolysates (PH) and trypsin inhibitor dietary inclusion on protein digestion/assimilation capacities, growth and survival. Four experimental diets were examined: C (control) I (supplemented with 750mg/kg soybean trypsin inhibitor (SBTI)) H (supplemented with 20% PH) and HI (supplemented with 20% PH and 750mg/kg SBTI). Protein hydrolysate supplementation gave significantly higher body mass than control at day 15 post-hatching. Unexpectedly, at day 30 and 60, fish administered diet HI (containing trypsin inhibitor) were heavier than the other groups. Suggested mechanisms are presented and discussed. The main conclusions of this study are that wolffish larval stage lasts roughly 15days and that juvenile growth is linked to proteolytic capacity, but also very likely to absorption capacity of peptides and amino acids. [Copyright &y& Elsevier]

The goal of this study was to determine the optimal stocking density for rearing juvenile spotted wolffish, Anarhichas minor (Olafsen), at two different sizes and assess the welfare status in relation to density. No major growth impairment was observed, although smaller fish (50–100 g) were significantly affected by density during the 120 days of the experiment, with final mean weights of 119.6 ± 11.6, 118.0 ± 5.8 and 88.7 ± 0.6 g for initial rearing densities of 10, 20 and 40 kg m−2 respectively. No effect of rearing density was seen for larger fish (100–160 g) during the 90 days of the experiment, with final mean weights of 160.2 ± 5.9, 159.7 ± 3.7 and 163.7 ± 11.5 g at fixed rearing densities of 20, 30 and 40 kg m−2 respectively. Our results suggest that the optimal rearing density of juvenile spotted wolffish is below 40 kg m−2 for smaller size fish (∼50–100 g) and probably ≥40 kg m−2 for the larger fish (100–160 g). Furthermore, it appears that the range of rearing density used did not have a significant effect on a selection of stress indicators (Na+ and K+ concentration, haematocrit, hepatosomatic index, total amount of plasma proteins and liver and muscle water content) and immunity response (plasma lysozyme activity) of juvenile spotted wolffish, making it a very tolerant species to crowding. [ABSTRACT FROM AUTHOR]

Abstract: The spotted wolffish (Anarhichas minor) is considered a promising species for coldwater aquaculture. It is a sedentary, bottom-dwelling fish which exhibits a calm and “non-stressed” behaviour in captivity. There are, however, no reports on the physiological responses to stressors in this species. In the present study we investigated primary (cortisol secretion) and secondary (glucose mobilization) responses to common aquaculture stressors like disturbance and hypoxia. Pre-stress plasma cortisol levels were within those generally considered representative for unstressed fish (~10ng ml−1), whereas basal glucose levels were unusually low (0.3–0.4mM). After exposure to a short, but severe disturbance challenge (emptying the tank of water for 10min), the increase in plasma cortisol level was slow and relatively weak, reaching a peak level of 25ng ml−1 4 to 8h after disturbance. When the fish were exposed to a gradually decreasing oxygen level in the tank, a significantly elevated plasma cortisol level (35ng ml−1) was seen in the fish that remained in the tank until oxygen saturation had decreased to 20% oxygen saturation after 2.5h. However, a two-fold higher plasma cortisol level (~70ng ml−1) was seen in all fish exposed to reduced oxygen levels (60, 40 and 20% oxygen saturation) after 3.5h recovery in normoxic water. Plasma glucose levels showed only moderate increases (~70%) following disturbance and hypoxia challenges. An in vivo injection of ACTH caused a strong elevation of plasma cortisol (peak level ~170ng ml−1), demonstrating a high capacity for interrenal steroidogenesis in the spotted wolffish. The slow and relatively weak cortisol response to stressors, and low plasma glucose levels, may relate to the sedentary lifestyle of the spotted wolffish. The stress-response is characterised by a passive (reactive) coping style, which is considered adaptive for farming of this species. [Copyright &y& Elsevier]

Abstract: Flat-bottom sea cages offer a cost-effective alternative for culture of bottom-dwelling fish. In an experiment lasting 428 days, spotted wolffish (Anarhichas minor Olafsen) grew equally fast when kept in tanks on land and in a flat-bottom sea cage at the same stocking density (increasing from 7.5 to 42 kg m−2 during the experiment) and same water temperature. A lag phase of 1 month with negative growth was observed following transfer from tanks to the sea cage. The results confirm that flat-bottom sea cages may be an option for culture of this species. [Copyright &y& Elsevier]

Abstract: Despite larval robustness characterized by the absence of metamorphosis and readiness for exogenous feeding based on commercial feed, the spotted wolffish (Anarhichas minor) displays highly variable survival at first-feeding. In this study, we investigated the use of three dietary concentrations of protein hydrolysates (PH, pre-digested proteins) (0, 10 and 20%) when newly hatched juvenile wolffish were held at three different rearing temperatures namely, 5, 8 and 12 °C to determine whether digestion of protein was a limiting factor for fish growth and survival. Final weights for fish at 5, 8,and 12 °C at day 60 were respectively 0.35, 1.19 and 2.02 g. Mean specific growth rates were 1.97, 4.01 and 4.88%/day and survival rates were 49.8, 53.4 and 33.2% respectively. No significant effects of PH were observed on growth or survival at any time during the experiment. However, as a general trend, fish survival was always higher when the diet contained 20% PH. We suggest that the degree of hydrolysis of the PH used may have been insufficient to induce specific digestive enzyme stimulation for promoting larval growth. Moreover, precocious ontogeny of the digestive system may have precluded any significant effect of using dietary protein hydrolysates. Our results are discussed in comparison with metamorphic species which characteristically display incomplete development of the digestive system. [Copyright &y& Elsevier]

Abstract: The Atlantic and spotted wolffish (Anarhichas lupus and A. minor, respectively) inhabit the cold waters of the northeast Atlantic Ocean. Although both species experience subzero water temperatures during winter, the Atlantic wolffish, which occupies shallower waters than the spotted wolffish, faces the greater threat of coming into contact with ice and freezing. This laboratory study was designed to determine whether these species differed in their abilities to resist freezing by examining the seasonal changes in blood plasma freezing points, antifreeze protein (AFP) activity and Na+ and Cl− concentrations when exposed to seasonally cycling water temperatures and photoperiod. The plasma of both species showed distinct seasonal cycles in all parameters with the highest values occurring during the winter. However, of the two species, only the Atlantic wolffish produced sufficient AFP to protect the fish down to the freezing point of seawater (−1.80 °C). The levels of AFP in the spotted wolffish were too low to impart any significant improvement in their resistance to freezing (approximately −0.8 °C). When wolffish were maintained in warm water under a seasonally changing photoperiod, the amplitude of the seasonal cycle in AFP activity was greatly reduced, indicating that low water temperatures are necessary to maximize plasma AFP levels. However, despite being maintained in warm water, plasma levels of AFP activity began to increase over summer values at the same time of year as did the fish exposed to seasonally changing water temperatures. This suggests that photoperiod plays a major role in the timing of the annual AFP cycle. [Copyright &y& Elsevier]

The article talks about the Gyrodactylus anarhichatis Mo & Lile (Monogenea: Gyrodactylidae) infection of Iceland's farmed spotted wolffish, Anarhichas minor Olafsen. It aims to investivate the potential pathogenicity of this parasite to farmed spotted wolffish. More information is given in the article.

Abstract: Atlantic cod and spotted wolffish fry were fed high-M alginate containing feed for 59 and 55days, respectively. During this period the fry showed a higher specific growth rate compared to controls. Uptake and distribution of alginate was studied by inclusion of the 125I-labelled molecule in the feed. The stomach and intestine contained the highest amount while the kidney, liver and spleen contained some, indicating that the alginate was taken up by the gut and transported to internal organs. Cod fry fed 0.06% and 0.1% high-M alginate showed a death rate of 51.4% and 53.3%, respectively. The lowest mortality, 48.1%, was found in fry fed 0.01% high-M alginate. Controls showed a mortality rate of 49.0%. Differences were, however, not statistically significant. Challenge of the immunostimulated fry (fed 0.02% and 0.06% alginate for 62days) with atypical Aeromonas salmonicida bacteria resulted in accumulated mortalities of 56% and 49%, respectively, 47days after infection. The group that received 0.06% alginate for a shorter period (47days) and then control feed until challenged, and the group that received alginate by bath reached a cumulative mortality of 59% and 60%, respectively. Lowest mortality (44%) was seen in the control group. Numerous microabscesses were found in both immunostimulated and control fish in secondary lamellae of the gills, haematopoietic tissues of the kidneys, the submucosa and mucosa of the intestine, the spleen, the liver and the myocardium of the heart. [Copyright &y& Elsevier]

The growth properties of juvenile spotted wolffish Anarhichas minor reared at 4, 6, 8 and 12° C, and a group reared under ‘temperature steps’, (T-step) i.e. with temperature reduced successively from 12 to 9 and 6° C were investigated. Growth rate and feed efficiency ration was significantly influenced by temperature and fish size. Overall growth rate was highest at 6° C (0·68% day−1) and lowest at 12° C (0·48% day−1), while the 4 and 8° C, and the T-step groups had similar overall growth rates, i.e. 0·59, 0·62 and 0·51% day−1 respectively. Optimal temperature for growth ( Topt G) and feed efficiency ratio (Topt FCE) decreased as fish size increased, indicating an ontogenetic reduction in Topt G and Topt FCE. The results suggest a Topt G of juvenile spotted wolffish in the size range 135–380 g, dropping from 7·9° C for 130–135 g to 6·6° C for 360–380 g juveniles. The Topt FCE dropped from 7·4° C for 120–150 g to 6·5° C for 300–380 g juveniles. A wider parabolic regression curve between growth, feed efficiency ratio and temperature as fish size increased, may indicate increased temperature tolerance with size. Individual growth rates varied greatly at all time periods within the experimental temperatures, but at the same time significant size rank correlations were maintained and this may indicate stable size hierarchies in juvenile spotted wolffish. [ABSTRACT FROM AUTHOR]

The article focuses on the electron microscopic study of endocytosis of indigenous bacteria and cell damage caused by Vibrio anguillarum in the foregut and hindgut of spotted wolffish Anarhichas minor. The authors suggested two mechanisms involved in bacterial translocation of indigenous gut bacteria. The difference in bacterial translocation between indigenous gut bacteria and pathogens has not been elucidated. The results reached a hypothesis that paracellular bacteria translocation is a secondary effect of detachment and loss of enterocytes and increased intracellular gap when the fry is exposed to V. anguillarum.

Abstract: Eggs and larvae from the spotted wolffish (Anarhichas minor Olafsen) were analysed to study the ontogeny of the complement component C3. By Western blotting experiments, the results revealed that C3 was present already in unfertilised eggs and that there was a decrease in C3 content towards hatch, whereas the results using reverse transcription–polymerase chain-reaction showed a continuous increase of C3 mRNA content towards hatch. These findings indicate a transfer of maternal complement component C3 to the offspring where the content of C3 decreases and the level of C3 mRNA increases towards hatch. Immunohistochemical experiments revealed that only the liver of spotted wolffish larvae contained complement component C3, suggesting that the liver is the main site of production. [Copyright &y& Elsevier]

Abstract: Non-specific mechanisms are important in the defence of all multicellular animals against pathogenic microorganisms. Macrophages and granulocytes play a central role in this respect. It is thus pertinent to develop methods for obtaining and cultivation of macrophages and assessing their functions in the spotted wolffish, a cold water species of current interest for the aquaculture industry. Kidney macrophages from the spotted wolffish (Anarhichas minor Olafsen) were isolated by density sedimentation using Percoll. The cells were highly phagocytic and possessed typical macrophage morphology evaluated by transmission and scanning electron microscopy. Using electron microscopic analysis, the size of the macrophages, collected from the Percoll density interface, was 5–9 μm. The viability in vitro was highest (87.1%) when the cells were kept at 13 °C with the addition of synthetic serum replacement (SSR-2) when measured 24 h after seeding. One day old cells were not significantly activated by addition of bacterial lipopolysaccharide (LPS) for 24 h when measured by reduction of nitroblue tetrazolium compared to control cells. The cells were negative in respect to synthesis and contents of complement component C3. [Copyright &y& Elsevier]

The first articially fertilized spotted wolffish eggs hatched only 10 years ago, and today the species is considered a very promising candidate species for cold water aquaculture in the North Atlantic. Recent research has focused on identifying key biological parameters in spotted wolffish aquaculture in order to establish a full production line for the species, and basic aspects of reproduction and larval development are now understood, controlled, and no longer limiting production. Spotted wolffish eggs (5-6 mm) have a protracted incubation period (800-1000 D°) and newly hatched individuals (20-25 mm) are well developed, with the only larval characteristic remaining being a relatively small yolk sac which is completely resorbed after 3-4 weeks. The species can be weaned directly on formulated feed, and high specific growth rates have been obtained in land-based culture facilities using shallow raceways. Adaptive immune responses are present early after hatching and few potential disease problems have been identified. Only one bacterial disease, atypical furunculosis, has been reported in farmed fish, but oil-emulsified vaccines have displayed efficient protection both in juvenile and adult fish. Ectoparasites may, however, constitute a problem during parts of the year when sea-water temperature increases. Optimal temperature for growth decreases with increasing fish size and is 10-12 °C for early juveniles and 4-6 °C for adult fish and broodstock. Spotted wolffish is a very robust species, and juveniles thrive at high densities and may be reared at a wide range of salinity levels. The species has further displayed a high tolerance to environmental changes in water quality parameters such as oxygen, carbon dioxide and un-ionized ammonia. Currently, the possibility of rearing spotted wolffish in flat-bottom net cages with shelves in the sea is being investigated. Preliminary results suggest that sea-based production may be a viable alternative to land-based rearing of the species in certain areas. [ABSTRACT FROM AUTHOR]

Farmed spotted wolffish (Anarhichas minor O.) is susceptible to infections with atypical Aeromonas salmonicida, and experimental oil-adjuvanted vaccines are previously reported to induce protection against the disease. For prophylactic treatment of juveniles, an immersion vaccine is the preferred alternative, and to test the efficacy of different bath vaccination strategies, a waterborne challenge model has to be established. Exposing the wolffish juveniles to 2×107–108 live bacteria ml−1 for 60–90 min resulted in 70–85% mortality. Bath vaccinating wolffish juveniles (∼25 mm) for 1 h in a bacterin containing 108 inactivated cells ml−1 did not induce specific antibody responses although the bacterial antigens were localised in the muscle, kidney and liver tissues. No protection was established in the bath vaccinated fish after waterborne challenge, nor did a second immersion boost improve the efficacy. For comparison, fish were i.p. vaccinated with an oil-emulsified bacterin and significant protection against the pathogen was induced both after injection and bath challenge. The protection was significantly higher when both the vaccine and the pathogen were administered by injection as compared to i.p. vaccination and challenge by bath. [Copyright &y& Elsevier]

To study the effect of early vaccination, wolffish juveniles of size 50 and 90 mm, respectively, were vaccinated with an oil-adjuvanted atypical A. salmonicida bacterin. Vaccination resulted in significant protection after challenge with the homologous bacterial strain and specific antibody responses were demonstrated against whole bacteria as well as purified A-layer protein and LPS by ELISA and Western blotting but individual variation in immune responses was apparent. The A-protein was the most immunogenic bacterial component.In addition, higher numbers of immunoglobulin producing cells were detected by in situ hybridisation in kidney and spleen of vaccinated fish compared to non-vaccinated fish. Plasma cells were also present in gut and gills in equal numbers irrespective of treatment. No plasma cells were found in the skin. Finally, the frequencies of expressed VHfamilies and CLisotypes of wolffish immunoglobulins were shown by PCR. The relative expression of the three variable regions of the Ig heavy chain and the three isotypes of the Ig light chain in the spotted wolffish spleen seemed to be unaffected by immunisation with a complex antigen like the A. salmonicida bacterin. [Copyright &y& Elsevier]

The spotted wolffish (Anarhichas minor Olafsen) is a species of the Perciformes, the most diverse and numerous order of all fish. A cDNA library from head kidney tissue was screened for immunoglobulin (Ig) heavy and light chain transcripts, and showed highest identity to Ig sequences from other perciform species. So far only one Ig class is described in spotted wolffish, but three VHfamilies were identified among the heavy chain transcripts. Highest diversity was located at the CDR3 region and demonstrates the importance of this gene element to the antibody repertoire. Two VLfamilies were identified among the light chain clones and three distinct isotypes were present. Use of polymerase chain reaction and in situ hybridisation techniques revealed individual variations in the relative expression of the three isotypes of light chains. [Copyright &y& Elsevier]

The complement component C3 was isolated from spotted wolffish (Anarhichas minor Olafsen) serum by polyethylene glycol precipitation, anion exchange chromatography and gel filtration. Silver staining in SDS-PAGE and rabbit anti-wolffish C3 antiserum used in Western blotting revealed that spotted wolffish C3 contains two polypeptide chains, Mr65 and 115 kDa, respectively. The high molecular weight α-chain of the C3 incorporated 14C-methylamine suggesting that it contained a reactive thioester group. The deduced amino acid sequence, after screening a liver cDNA expression library, showed that the wolffish C3 contained key amino acids for binding C3 convertase, factor H, I and properdin. Also, high degree of homology to other vertebrate C3 was found in the β–α junction site. Phylogenetic tree analysis indicated that the Japanese flounder and spotted wolffish that belong to order pleuronectiformes and perciformes, respectively, are phylogenetically close species. Immunohistochemical experiments showed that liver hepatocytes and blood contained C3, and in situ hybridisation experiments revealed that liver hepatocytes expressed C3. [Copyright &y& Elsevier]

The effects of dissolved oxygen (DO) concentration combined with sublethal doses of ammonia on growth and food conversion efficiency (FCE) were investigated in juvenile spotted wolffish. Fish with a mean (S.D.) initial weight of 95.4 (18.0) g were reared in shallow raceways (initial stocking density 10 kg m−2) at normoxic (9.6 mg l−1) and hyperoxic (14.5 mg l−1) conditions, and also under both normoxia and hyperoxia with an additional sublethal concentration of unionised ammonia (0.17 mg NH3 l−1) added, for 8 weeks at 8 °C. There was an interacting effect of oxygen saturation and ammonia level on growth, as growth rate was significantly higher in the hyperoxic/NH3 group compared to the normoxic/NH3 group (P<0.001), suggesting that hyperoxic conditions may increase tolerance to unionised ammonia in spotted wolffish. At the end of the experiment, no difference in mean weight was found between fish reared at normoxic and hyperoxic conditions without added ammonia, whereas mean weight was reduced under normoxia with added ammonia as compared to normoxia without added ammonia. Mean daily oxygen consumption was investigated in the non-ammonia groups at both normoxic and hyperoxic conditions, and O2 consumption was significantly higher (P<0.001) under hyperoxic (100.5 mg O2 kg−1 h−1) compared to normoxic (79.9 mg O2 kg−1 h−1) conditions. A clear diurnal rhythm in the O2 consumption pattern was seen at both O2 saturations, with the most striking observation being a dramatic increase in the morning when the light was turned on. Overall, our results support earlier findings of a high capacity to adapt to variations in environmental water quality parameters, and to restore normal growth rate in the spotted wolffish. [Copyright &y& Elsevier]

The presence of immunocompetent cells was studied in the larval and adult stages of the spotted wolffish, Anarhichas minor. In situ hybridization with a probe complementary to the secretory Igμ-chain was used to localize immunoglobulin producing cells or plasma cells in organs from adult fish and the appearance of these cells in lymphoid tissues during juvenile development. Plasma cells were located in pronephros, spleen, gut, gills and skin of adult wolffish. In juveniles, the first plasma cells were detected in the kidney 1 week post-hatching and the appearance in other lymphoid organs was in the order spleen, gut and thymus. No plasma cells were detected in skin and gills during the sampling period of juveniles (<10 cm). Our study confirmed that plasma cells are present in both the systemic and mucosal compartments of adult fish but during ontogeny there is an earlier appearance of plasma cells in the gut compared to gill and skin compartments. [Copyright &y& Elsevier]

Kininogens are multifunctional proteins found so far mainly in mammals. They carry vasoactive kinins as well as participate in defense, blood coagulation and the acute phase response. In this study, novel kininogens were isolated from Atlantic cod (Gadus morhua L.) and spotted wolffish(Anarhichas minor ) by papain-affinity chromatography. The molecular mass of cod kininogen determined by MALDI-TOF mass spectrometry to be 51.0 kDa and it had pI values of 3.6, 3.9 and 4.4. The molecular mass of wolffish kininogen was 45.8 kDa and it had pI values of 4.1, 4.3, 4.35 and 4.4. Partial amino-acid sequences determined from both kininogens showed clear homology with previously determined kininogen sequences. Both kininogens were found to inhibit cysteine proteinases like papain and ficin but they had no effect on trypsin, a serine proteinase. Wolffish kininogen carried α2,3-sialylated biantennary and triantennary N-glycans with extensive sialic acid O-acetylation. Cod kininogen carried similar glycan structures but about 1/3 of its glycans carried sulfate at their N -acetylglucosamine units. [ABSTRACT FROM AUTHOR]

The atypical strains of Aeromonas salmonicida isolated from spotted wolffish Anarhichas minor O., show biochemical and molecular variations, but it is not known whether these variations affect bacterial virulence or antigenicity of possible protective antigens. In this study, protection against atypical furunculosis was compared in spotted wolffish vaccinated with monovalent or multivalent vaccines containing different strains of atypical A. salmonicida. In addition, these vaccines were compared to a vaccine containing an atypical A. salmonicida isolate from Atlantic halibut, Hippoglossus hippoglossus L. Vaccinated fish were challenged with three different A. salmonicida strains from spotted wolffish. Significant protection was obtained against both homologous and heterologous challenge. The best protection, regardless of strain used for challenge, was obtained with the multivalent vaccine composed of three different bacterial strains. Also, some of the monovalent vaccines resulted in significant protection, but they seemed to differ in protection against one particular strain of atypical A. salmonicida. In addition, significant cross-protection was observed with a vaccine containing a strain isolated from halibut, while a commercial salmon vaccine against furunculosis based on A. salmonicida subsp. salmonicida, failed to protect against an atypical strain from spotted wolffish. [Copyright &y& Elsevier]

Eggs of spotted wolffish ( Anarhichas minor Olafsen)were incubated at constant 4, 6 and 8 °C, and disinfected withglutaric dialdehyde (150 p.p.m. for 5 min) once ortwice a month during two thirds of the incubation period, to prevent growth ofmicroorganisms. Hatching of apparently normal larvae started earlier when eggswere disinfected twice a month compared to once a month at all incubationtemperature regimes. The time to 50% hatch was 900 and 920 day-degrees (16 and16,5 weeks) at 8 °C, 835 and 880 day-degrees (20 and 21 weeks)at 6 °C and 725 and 800 day-degrees (26 and 28,5 weeks) at 4°C, in the egg groups disinfected twice or once a month,respectively. The best survival until hatching was noted when eggs weredisinfected twice a month and incubated at 6 and 8 °C.Survivalwas very low at 4 °C. Prematurely hatched larvae wereregistered in all egg groups disinfected twice a month and the highestfrequencywas noted in the 8 °C groups. The larval weight at normalhatching in the 6 and 8 °C groups was negatively correlatedwith incubation temperature and intervals of disinfection during the incubationperiod, but after 42 days feeding with live feed (unenriched Artemia) the weights of the larvae were not significantlydifferent. The specific growth rates of the larvae from the eggs incubated at 6°C and 8 °C were 3.0% and 3.2%, respectively.The mean survival of larvae was between 88% and 96% at 42 days post-hatching.Young wolffish originating from the 6 °C incubation groupsshowed lowest mortality. [ABSTRACT FROM AUTHOR]

Simple Summary: Spotted wolffish, a cold-water fish species, is a high potential candidate marine fish species for the diversification of the aquaculture sector in Norway, Canada, and Iceland. A review of the state of advancement of all aspects of its reproduction is proposed. Species-specific life-history and reproduction traits are discussed in parallel with relevant information originating from past R&D activities and the current state of knowledge regarding rearing environmental conditions and practices possibly affecting broodstock performances. The first artificially fertilized spotted wolffish (Anarhichas minor) eggs hatched in Norway in the mid-1990s as this species was considered by Norwegian authorities to be a top candidate species for cold-water aquaculture in the North Atlantic regions. Previous research conducted in Norway (since 1992) and Canada (since 2000), focused on identifying key biological parameters for spotted wolffish cultivation which led, respectively, to the rapid establishment of a full commercial production line in northern Norway, while Québec (Canada) is witnessing its first privately driven initiative to establish commercial production of spotted wolffish on its territory. The control of reproduction can be viewed as a major requirement to achieve the development of performant strains using genetic selection tools and/or all-year-round production to bring about maximal productivity and synchronization among a given captive population. Although the basic reproduction aspects are more understood and controlled there are still some challenges remaining involving broodstock and upscaling of operations that limit the achievement of a standardized production at the commercial level. Quality of gametes is still considered a major constraint and it can be affected by multiple factors including nutrition, environmental conditions, handling practices, and welfare status. Internal insemination/fertilization and the protracted incubation period are challenging as well as the establishment of a health monitoring program to secure large-scale operations. The profound progress achieved in the control of reproduction, sperm handling, and cryopreservation methods for this species is presented and discussed. In this review, we also go into detail over the full range of up-to-date cultivation practices involving broodstock and identify areas that could benefit from additional research efforts (i.e., broodstock nutrition, health and welfare, scaling-up egg and larval production, genetics, and development of selective breeding programs). [ABSTRACT FROM AUTHOR]