Exploring the Effects of Acute Stress Exposure on Lumpfish Plasma and Liver Biomarkers.

This study aimed to expand knowledge on lumpfish stress physiology by investigating the effects of acute stress on primary (i.e., cortisol) and secondary (e.g., metabolites) stress responses, as well as oxidative stress biomarkers, from stress exposure to a recovery phase. The results showed that the lumpfish physiological response to 1 min air exposure is mild, in line with recent studies, and comparable to that described for white sturgeons. Cortisol seems to be the most reliable acute stress biomarker in lumpfish, with a significant increase in plasma 30 min after stress exposure, returning to resting levels 2 h after exposure. In contrast, glucose and lactate were not significantly altered by short-term air exposure. Effects on hepatic energy mobilisation were also detected following the acute stress. This study showed that acute 1 min air exposure seems tolerable, allowing a swift recovery. However, more studies on the impacts of air exposure and repeated acute stressors on lumpfish stress and immune responses are required to develop industry standards for lumpfish health and welfare monitoring.

Mitogenomics of the suborder Cottoidei (Teleostei: Perciformes): Improved assemblies, mitogenome features, phylogeny, and ecological implications.

We determined the mitogenome of Cyclopterus lumpus using a hybrid sequencing approach, and another four closely related species in the Liparidae based on available next-generation sequence data. We found that the mitogenome of C. lumpus was 17,266 bp in length, where the length and organisation were comparable to those reported for cottoids. However, we found a GC-homopolymer region in the intergenic space between tRNALeu2 and ND1 in liparids and cyclopterids. Phylogenetic reconstruction confirmed the monophyly of infraorders and firmly supported a sister-group relationship between Cyclopteridae and Liparidae. Purifying selection was the predominant force in the evolution of cottoid mitogenomes. There was significant evidence of relaxed selective pressures along the lineage of deep-sea fish, while selection was intensified in the freshwater lineage. Overall, our analysis provides a necessary expansion in the availability of mitogenomic sequences and sheds light on mitogenomic adaptation in Cottoidei fish inhabiting different aquatic environments.

The effect of temperature on growth performance and aerobic metabolic scope in Arctic charr, Salvelinus alpinus (L.).

In recent years, Arctic char populations in Iceland have declined and the objective of this experiment was to throw further light on these changes by examining the effect of temperature (5, 9, 13, 17, and 21 °C) on the survival, growth rate, metabolism, and physiological indices of juvenile Arctic charr (initial mean body mass 4.02 ± 0.8 g). Mortality was 60% at 21 °C while at lower temperatures it was below 5%. However, Arctic charr populations in Iceland are declining in locations where the ambient temperature is lower, suggesting that other factors may be more important in determining the abundance of the species. The optimum temperature for growth was near 14 °C. The growth rate was progressively reduced at supra-optimum temperatures with almost no growth at 21 °C. Indicators of energy reserves: condition factor, relative intestinal mass, and hepatosomatic index are all consistent with reduced feed intake at supra-optimum temperatures. The standard and maximum metabolic rate (SMR; MMR), as well as the aerobic scope for activity (AS), were maximum at 13 °C. The routine metabolic rate (RMR) increased exponentially with temperature and, at T21, it was equal to the MMR suggesting, that the RMR was limited by the MMR. Moreover, increased heart- and gill mass at 21 °C are consistent with increased stress on the cardiovascular system. These findings are in keeping with the OCLTT hypothesis that the thermal tolerance of fish is limited by the capacity of the cardiovascular system to deliver oxygen and support metabolism. Taken together, the results of this experiment suggest, that growth rate is reduced at supra-optimum temperatures because of reduced energy intake, increased metabolic demand, and limitations in the capacity of the cardiovascular system to support metabolic rate at high temperatures. At lower temperatures, growth does not appear to be limited by the AS.

Genome- and transcriptome-derived microsatellite loci in lumpfish Cyclopterus lumpus: molecular tools for aquaculture, conservation and fisheries management.

The lumpfish Cyclopterus lumpus is commercially exploited in numerous areas of its range in the North Atlantic Ocean, and is important in salmonid aquaculture as a biological agent for controlling sea lice. Despite the economic importance, few genetic resources for downstream applications, such as linkage mapping, parentage analysis, marker-assisted selection (MAS), quantitative trait loci (QTL) analysis, and assessing adaptive genetic diversity are currently available for the species. Here, we identify both genome- and transcriptome-derived microsatellites loci from C. lumpus to facilitate such applications. Across 2,346 genomic contigs, we detected a total of 3,067 microsatellite loci, of which 723 were the most suitable ones for primer design. From 116,555 transcriptomic unigenes, we identified a total of 231,556 microsatellite loci, which may indicate a high coverage of the available STRs. Out of these, primer pairs could only be designed for 6,203 loci. Dinucleotide repeats accounted for 89 percent and 52 percent of the genome- and transcriptome-derived microsatellites, respectively. The genetic composition of the dominant repeat motif types showed differences from other investigated fish species. In the genome-derived microsatellites AC/GT (67.8 percent), followed by AG/CT (15.1 percent) and AT/AT (5.6 percent) were the major motifs. Transcriptome-derived microsatellites showed also most dominantly the AC/GT repeat motif (33 percent), followed by A/T (26.6 percent) and AG/CT (11 percent). Functional annotation of microsatellite-containing transcriptomic sequences showed that the majority of the expressed sequence tags encode proteins involved in cellular and metabolic processes, binding activity and catalytic reactions. Importantly, STRs linked to genes involved in immune system process, growth, locomotion and reproduction were discovered in the present study. The extensive genomic marker information reported here will facilitate molecular ecology studies, conservation initiatives and will benefit many aspects of the breeding programmes of C. lumpus.

Is Habituation Measurable in Lumpfish Cyclopterus lumpus When Used as Cleaner Fish in Atlantic Salmon Salmo salar Aquaculture?

To investigate how lumpfish interact in Atlantic salmon aquaculture, physiological stress responses and changes in behaviour were analysed in experienced and naive lumpfish. Experienced lumpfish (30.2 ± 7.93 g, mean ± SD) coexisted with a commercial scale production unit of Atlantic salmon (1258.5 ± 152.12 g) for 30 to 60 days, while naive lumpfish (38.2 ± 12.37 g) were kept with conspecifics only. Ten trials from each background were tested. For each trial, 10 lumpfish were tagged and transferred to a video monitored experimental tank (2 × 2 × 0.7 m). In each trial, swimming behaviour was mapped for all lumpfish every 60 s in 20 min, 10 min before, and 10 min after the introduction of four Atlantic salmon. Naive lumpfish expressed significantly increased burst swimming activity and maintained longer interspecific distance to Atlantic salmon in comparison with experienced fish. In addition, mean plasma cortisol levels were significantly elevated in naive fish after exposure to Atlantic salmon. We argue that naive lumpfish expressed innate physiological and behavioural stress responses during first encounter with Atlantic salmon, while reduced responses in experienced individuals indicated habituation. The effect from behavioural and physiological stress in newly deployed naive lumpfish-before and during habituation-should be taken account for when lumpfish are introduced in commercial sea cages to improve welfare for the species. In addition, we suggest that habituation could be applicable during the rearing phase to moderate the transition from a simple tank environment with conspecifics only to interspecies interaction with Atlantic salmon in sea cages.

Modelling sea lice control by lumpfish on Atlantic salmon farms: interactions with mate limitation, temperature and treatment rules.

Atlantic salmon farming is one of the largest aquaculture sectors in the world. A major impact on farm economics, fish welfare and, potentially, nearby wild salmonid populations, is the sea louse ectoparasite Lepeophtheirus salmonis. Sea louse infestations are most often controlled through application of chemicals, but in most farming regions, sea lice have evolved resistance to the small set of available chemicals. Therefore, alternative treatment methodologies are becoming more widely used. One increasingly common alternative treatment involves the co-culture of farmed salmon with cleaner fish, which prey on sea lice. However, despite their wide use, little is understood about the situations in which cleaner fish are most effective. For example, previous work suggests that a low parasite density results in sea lice finding it difficult to acquire mates, reducing fecundity and population growth. Other work suggests that environmental conditions such as temperature and external sea louse pressure have substantial impact on this mate limitation threshold and may even remove the effect entirely. We used an Agent-Based Model (ABM) to simulate cleaner fish on a salmon farm to explore interactions between sea louse mating behaviour, cleaner fish feeding rate, temperature and external sea louse pressure. We found that sea louse mating has a substantial effect on sea louse infestations under a variety of environmental conditions. Our results suggest that cleaner fish can control sea louse infestations most effectively by maintaining the population below critical density thresholds.

The use of photoperiods to provide year round spawning in lumpfish Cyclopterus lumpus.

In order to provide year round spawning broodstock, lumpfish (initial size 746 g and 24.9 cm) were reared under four different photoperiod regimes from January 2017 to July 2018. One group was reared under simulated natural photoperiod (LDN, control group) for Tromsø (70°N). The second group was transferred to continuous light (LD240) on 30 January 2017 and reared at LD24:0 throughout the trial period. Two compressed and phase advanced photoperiods were also established. Both groups were moved from LDN to LD24:0 on 30 January 2017, and after that reared at compressed natural photoperiods where the annual photoperiod was compressed down to six months (L6) or nine months (L9) for the duration of the study. Spawning time was shifted in both compressed groups during both years of the study. Spawning activity in the second year of the study was higher and followed more closely the expected spawning period in the compressed and the LDN groups. Spawning in the LD240 group was spread out over the experimental period with no distinct peak in spawning. A seasonal and pronounced drop in condition factor was found for females in the L9, L6 and the LDN groups. This post-spawning loss in condition was closely related to the spawning activity of each group. The current findings suggest that photoperiod has a strong influence on the timing of lumpfish maturation and can be used as an efficient and inexpensive tool to secure lumpfish reproduction operations i.e. year-round supply of egg and milt and/or timing with optimal temperature regimes.

Ontogenetic effects of diet during early development on growth performance, myosin mRNA expression and metabolic enzyme activity in Atlantic cod juveniles reared at different salinities.

This study investigates the effect of diet during early development on growth and metabolic capacity in the juvenile stage of Atlantic cod. Growth in three groups of Atlantic cod juveniles (10-70 g) was measured at two salinities (15 per thousand or 32 per thousand) in combination with two temperatures (10 degrees C or 14 degrees C). Groups of cod from a single egg batch differed by having been fed with rotifers (R) or natural zooplankton (Z) during the first 36 days post hatch. A third group was fed zooplankton from 1 to 22 dph, after which diet changed to rotifers from 22 to 36 dph (ZRZ). All fish were weaned at 36 dph. Juveniles from the Z and ZRZ groups performed equally well under all experimental conditions, but fish that had received rotifers as a larval diet showed overall significantly lower growth rates. Growth was significantly enhanced by reduced salinity. Metabolic enzyme activity and relative myosin mRNA expression levels were not affected by larval diet. Muscle AAT and MDH were affected by salinity while these enzymes in liver tissue were affected by the interaction between salinity and temperature. Metabolic enzymes were stronger correlated with fish size than growth rates. Our results indicate that larval diet has a pronounced effect on juvenile growth rates under varying environmental conditions as optimal larval diet (zooplankton) increased juvenile growth rates significantly. Metabolic enzyme activity and relative myosin mRNA expression were not affected by larval history, which suggests that the persisting juvenile growth difference is not a result of differing metabolic capacity.

Plasma insulin-like growth factor-I concentrations and growth in juvenile halibut (Hippoglossus hippoglossus): effects of photoperiods and feeding regimes.

The effects of photoperiod and feeding regimes on plasma IGF-I levels and their relationship with growth rate of juvenile halibut (initial mean weight 364 g) were investigated by rearing fish under five different photoperiod regimes and two feeding regimes for 14 months. The entire photoperiod experiment was divided into 3 phases where the fish in each phase were exposed to either natural photoperiod (N), stimulated photoperiod with long day and short night (S) or continuous light (L). Thus, the following five photoperiod combinations were tested: a) Control group (NNN) b) Group 2A (NLN) c) Group 2B (NNL) d) Long day-natural group (SNN) e) Production group (LNN). In addition, the Control group was split into two parts and fed according to two different feeding regimes: a) Continuous fed group: Fish fed every day. b) Starvation/re-fed group: Fish were starved for 5 weeks and then re-fed for 10 weeks, and the treatment repeated during the whole experimental period. The analyses of IGF-I were performed from individually tagged fish in all groups in September 2005 and March 2006. In order to test how rapidly starvation affects circulating IGF-I levels samples were taken from the Starvation/re-fed group after a 10 days starvation (September) and immediately after 10 weeks of feeding (March). A significant relationship between IGF-I levels and individual growth in the preceding period and photoperiod and starvation treatment was found on both occasions. In conclusion, the present study indicates that plasma IGF-I levels are correlated to growth in Atlantic halibut, and affected by photoperiod treatment or compensatory growth during re-feeding. Correlation between individual growth rate and IGF-I levels was low, but significant, highlighting the complexity of how environmental factors affect the endocrine and physiological regulation of growth in fish.

New haemoglobin genotypes in Atlantic cod, Gadus morhua: possible relation with growth.

In a preliminary study, 121 individually tagged juvenile Atlantic cod (Gadus morhua) were classified according to their haemoglobin genotypes into four groups, i.e., two main haemoglobin genotypes [Hb-I(1/2), Hb-I(2/2)] and two sub-types [Hb-I(1/2b), Hb-I(2/2b)], and reared for 3 months at 10 degrees C, 13 degrees C and T-step (fish reared at 16 degrees C and then subsequently moved to 13 and later to 10 degrees C). Overall growth rates across temperatures were 10% and 19% higher in the Hb-I(2/2b), Hb-I(1/2b) sub-types compared to corresponding Hb-I(2/2) and Hb-I(1/2) main types, respectively. Individual growth rate trajectories varied between the genotypes at all temperatures studied. Our study indicates that under certain environmental conditions higher growth in the two sub-types compared to the main genotypes could be expected. This may indicate difference in other physiological characters not studied here, but seen in previous studies, i.e., oxygen affinity and competitive performance.

Environment affects stress in exercised turbot.

We investigated the interaction of water temperature (10, 18 and 22 degrees C) and salinity (33.5 and 15 per thousand ) on the stress response of juvenile turbot. At each temperature/salinity combination, fish were subjected to 10 min enforced exercise. This induced a moderate stress response, which differed at the various temperature and salinity combinations. High temperatures caused more rapid increases in plasma cortisol and glucose, larger and more rapid increases in plasma lactate levels, which were also influenced by body weight, and a faster recovery in plasma Na(+) and Cl(-). Low salinity ameliorated cortisol responses at low but not at high temperatures. The magnitude of ionic disturbance was reduced at 15 per thousand. Plasma K(+) did not change at any temperature or salinity. The stress response involved activation of the brain-pituitary-interrenal axis, as indicated by the cortisol elevations. The low magnitude of glucose responses, the mild Na(+) and Cl(-) disturbances, and the lacking K(+)-responses indicated mild activation of the brain-sympathetic-chromaffin cell axis, and hence a low release of catecholamines, which seemed though to occur to a higher extent at higher temperatures. The relatively low catecholaminergic response of turbot may be linked to their inactive sedentary lifestyle. The higher responsiveness at higher water temperatures may reflect a higher overall adaptive capacity.

HPV subtypes and immunological parameters of cervical cancer in Iceland during two time periods, 1958-1960 and 1995-1996.

OBJECTIVE: Cervical cancer is a disease caused in part by an infection with an oncogenic subtype of human papillomavirus (HPV). In this study we analysed all cervical cancer samples diagnosed in Iceland during two periods, 1958-1960 and 1995-1996, and asked whether significant changes in viral or immunological parameters had occurred over a period that spanned both significant changes in sexual attitude and the implementation of organized screening for cervical cancer. METHODS: Samples from 47 patients (46 squamous cell carcinomas (SCC) and 1 adenosquamous carcinoma (ASC)) in the first period and 30 patients (20 SCC, 4 ASC, and 6 adenocarcinomas (AC)) in the later period were analysed for viral subtype and expression of Fas, FasL, MHC class I, p53 and apoptosis. RESULTS: AC and ASC are proportionately much more common today than 40 years ago (30% vs 2%). The distribution of HPV in cervical cancer is similar in both periods, with HPV16 found in 75% and HPV18 in 13% of cases. Other HPV types found were 31,33,45, and 59. No significant differences were found in the immunological profiles of tumors from the two periods except that a higher fraction of SCC in the later period stained positive for FasL. When SCC are compared with AC/ASC, the latter have less expression of MHC class I, less expression of Fas, and stronger FasL expression. CONCLUSIONS: AC/ASC tumors show some immunological features that suggest that they are more resistant to immune attack than SCC.