The purpose of this study was to apply oxygen and carbon stable isotope microsampling techniques to the seasonal banding of the otoliths of the Atlantic cod, Gadus morhu~ in order to determine their potential use in reconstructing the environmental conditions experienced by cod. A reconstruction of the changes in environmental temperature is seen as particularly applicable to the determination of cod migration routes based on the known temperature profile of the North Atlantic. Transverse thin sections of six otoliths from cod taken off the coast of Atlantic Canada were prepared using standard methods. Material was sampled from each semiannual band of the otoliths and run on a Finnigan MAT 251 ratio mass spectrometer to determine 0180 and 013C. The results for 0180 indicated that the otoliths had an approximate initial value of either 1.4%o or l.O%o for sample material taken from the nucleus. The 0180 signal was characterized by oscillating values in which sample material from hyaline bands corresponded predominantly with the troughs and sample material from the opaque bands corresponded with the peaks. The average range of 0180 was found to be 0.87%o corresponding to a temperature range of 3.6°C which was within the expected 3 to 4 °C seasonal average temperature shift experienced by cod. Seasonal cycling was apparent in three of the otoliths, with semi-annual values alternating between high opaque bands and low hyaline bands. It was concluded that seasonal temperature changes due to migration from offshore to inshore waters are recorded in the 0180 signal and that future sampling should attempt to resolve several samples within each seasonal band in order to resolve the migratory changes in temperature on an sub-annual basis. It is also suggested that experiments be carried out to determine the species specific 0180 versus temperature relationship for cod to make accurate interpretation of the data possible. The o13C signal was found to be characterized by an increasing logarithmic trend in o13C. The 013C signal was observed to increase over the first three years of growth and to level off at age 4 and fluctuate at a value close to O%o. The otoliths of cod were found to contain a high proportion of inorganic carbon with a minimum 013C value of -4.53%o and a maximum of 0.23%o. It was suggested that the initial increase in 013C was a product of a combination of factors affecting metabolic rate and therefore, indirectly, the amount of metabolically derived carbon circulating in the blood. It was concluded that further research into the growth and development of cod was necessary in order to reach a comprehensive understanding of the biological processes responsible for the observed trends in 013C. 0180 was plotted against 013C for samples from growth bands of age four or greater and found to have a positive correlation with a slope of 0.269 (S.E. 0.049) and an r-squared of 0.537, P < 0.0001 when the results for otolith 176 were excluded due an anomalous trend in that data set. Two hypotheses were suggested to account for the observed positive correlation, a metabolic/temperature effect, and a depth/temperature effect. It was concluded that, with further research into the controlling factors behind the fluctuations in 013C, the 013C signal may provide a second source of information with regards to changes in habitat and environmental characteristics over the life time of the cod. It is suggested that a study be carried out to determine the amount of metabolic carbon incorporated into the otolith in order to clarify the observed trends. / Thesis / Candidate in Philosophy

Fiskbestånden i Östersjön har sedan början av 1980-talet genomgått en omfattande förändring. Stora predatorer, som torsk (Gadus morhua) och abborre (Perca fluviatilis) har minskat i antal medan det finns tecken på att storspigg (Gasterosteus aculeatus) har ökat. Denna sannolika ökning av storspigg skulle kunna ha att göra med minskningen av både torsk och abborre som predatorer i de kustnära tångbältena. För att undersöka detta utfördes ett mesokosmexperiment där torskens och abborrens förmåga att predera på storspigg undersöktes, både i ljus och mörker. Hypoteserna som arbetet utgick ifrån var att torsk var en bättre predator i mörker än i ljus och att torsk var en bättre predator än abborre i mörker. Resultaten visar på att torsk är den bättre predatorn i både ljus och mörker, samt att den är lika framgångsrik i mörker som i ljus. Detta tyder på att torskens försvinnande skulle kunna vara en bidragande faktor till ökningen av storspigg. Ytterligare visades abborren vara en mer framgångsrik predator i mörker än i ljus, detta trots att den alltid anges som en visuell predator. Detta visar att även minskningen av abborre skulle kunna vara en anledning till att storspiggen ökar.

The persistence of a species in the face of environmental change is a function of the extent to which populations respond differently to changes in their environment and the spatial correspondence between the scale of disturbance and the scale of adaptation. The pattern by which a population, or genotype, expresses a range of phenotypes across an environmental gradient is called a norm of reaction. The level of phenotypic plasticity displayed within a population (i.e. the slope of the reaction norm) reflects the short-term response of a population to environmental change while variation in reaction norm slopes among populations reflects the spatial scale of variation in these responses. Using a reaction norm framework, I examined the spatial scale of genetic variation in plasticity for life-history traits in Atlantic cod (Gadus morhua), a marine fish of global biological and socioeconomic importance. Through common-garden experiments, I found evidence of both adaptive and non-adaptive plasticity for larval growth rate and survival in two cod populations that experience contrasting thermal environments in nature. A comparison of these reaction norms with those of four cod populations studied previously revealed significant genetic divergence in adaptive traits at a smaller spatial scale than has previously been shown for a marine fish with no apparent physical barriers to gene flow (<250 km). This fine-scale genetic structure is likely the result of populations being locally adapted to seasonal changes in temperature during the larval stage caused by differences in spawning times and may be maintained by behavioural barriers to gene flow. Implications of variation in life-history trait plasticity to fisheries management in the face of predicted changes in climate are discussed.

The populations of Atlantic cod (Gadus morhua) in the Baltic Sea has been heavily exploited for decades, with fishing mortalities close to, and for several occasions, even above one. The larger part of the spawning stock biomass is consequently being removed each year. The issue of fisheries induced evolution (FIE) has been gaining attention from researchers lately. The selection pressure driving this evolution is powered by a connection between high mortality rates and heritable traits. The fishing in the Baltic Sea is mainly performed with size selective gear that can impose selection on traits like size-at-age or size-at-maturity. In this review I show how FIE may affect the Baltic cod towards decreased size-at-age/maturity and how size selective fishing on stocks at low levels can increase the inherent instability of the population as well as deprave the Baltic Sea of ecosystemic services from cod.

Sexual maturation during commercial culture of Atlantic cod (Gadus morhua L.) represents a significant production bottleneck restricting the profitability of the industry. Such problems in other species have traditionally been addressed by artificial manipulation of photoperiod cycles, however little research exists in this field in cod. This thesis therefore investigates the interactions between artificial photoperiod manipulation, sexual maturation and somatic growth in this species. In the first experiment, populations of Atlantic cod (hatched, spring 1999) were maintained on either a simulated natural photoperiod (SNP) or continuous illumination (LL) from approximately 15 months post hatch (MPH) (July 2000) in an enclosed tank system. Growth performance was recorded monthly along with observations of reproductive activity over the subsequent 2 years (up to July 2002). At both 2 and 3 years of age the entire population raised under SNP matured and spawned, during which time mean weight reduced by 13% and 24% respectively. No spawning individuals were recorded at 2 years of age in the LL population and only 18% were observed to spawn at 3. However, observations of both changes in gonadal morphology (observed via ultrasound scanning) and a suppression in growth rate at 2 years of age in the LL population alluded to a maturation “dummy run” regulated by an endogenous clock. Despite this phenomenon, the LL treatment realised a 39% and 43% improvement in wet weight following 1 and 2 years of exposure to LL respectively. When the diel cycle of plasma melatonin was compared between the treatments in February 2001 (23MPH) the SNP population displayed an A-profile diel rhythm ranging between 20 and 50 pg/ml while the LL treatment did not display any rhythm. In the second experiment of this work, two populations of cod (hatched, spring 2001) were reared in commercial open cage systems, one of which experienced continuous additional artificial illumination between July 2002 (15MPH) and October 2003 (30MPH) provided by four, 400W submerged lighting units. Growth and maturation were assessed in both populations throughout. In March 2003 (24MPH) it was apparent that spawning individuals were present in both the SNP and LL populations though a significantly lower number of spawning individuals in the LL treatment suggested that the peak in spawning activity was delayed by about 1 to 2 months. With both populations apparently maturing at 2 years of age, there was no significant difference in weight between the populations at the end of the trial. In comparison to experiments I and IV of this work, these results would suggest that in comparison to salmonids for example, Atlantic cod appear to have a heightened sensitivity to light allowing individuals to differentiate the ambient photoperiod signal from the application of continuous artificial light. In the third experiment, 6 populations of approximately 20 tagged individuals (hatched spring 1999) were maintained, from December 2000 to July 2002, under either SNP, LL or one of four, out of season “square wave” photoperiod regimes (repeating cycles with a 12 month period, consisting of a 6 month window of LL followed by six months of short day lengths [SD, 7L:17D] which had been staggered to start over a six month period). Each individual was monitored monthly for maturation status. Out of season “square wave” photoperiods were demonstrated to successfully entrain maturation and hence significantly alter the spawning profiles in these populations. Application of LL from December 2000 failed to inhibit maturation in the spring of 2001 and, in fact, advanced the spawning season by 1 month while those that experienced SD from the same date showed significant extension of the subsequent spawning season. Interestingly, the males maintained on LL throughout the experiment matured both in the spring of 2001 and one year later in the spring of 2002 while females under the same treatment only matured and spawned in 2001. In the fourth experiment, a total of 830 tagged individuals were raised either under SNP or one of 7 photoperiod treatments, consisting of 5 groups transferred from SNP to LL at 3 monthly intervals between 6 and 18 MPH where they remained and a further two groups maintained on LL from 6 to 15MPH and 6 to 21 MPH respectively before being returned to SNP. Both the gonadic and somatic axes were monitored at the physiological and endocrinological level at three monthly intervals from 6 to 27 MPH. The results demonstrated that it is the falling autumnal photoperiod signal after the summer solstice, more specifically after October, that is responsible for recruiting individuals to enter the sexual maturation cycle. Furthermore, in all treatments where this signal was masked i.e. those which experienced LL starting at or prior to 15MPH, except for some restricted spermatogenic activity in the males testis observed at 27MPH, there was no significant reproductive activity and growth was improved by up to 60% at 27 MPH. While providing evidence for direct photic stimulation of somatic growth, the growth results were also correlated with the measurement of plasma IGF-I and demonstrated its potential as a tool to assess growth rates in the species. Plasma melatonin measured at 15MPH, as in experiment I, was suppressed in all populations which were under LL photoperiods. By identifying the photoperiod “window of opportunity” which recruits individuals into the sexual maturation cycle, this work was able to conclude that the application of LL from the summer solstice prior to maturation is the most efficient photoperiod strategy to be adopted by the aquaculture industry to realise maximum growth potential from their cultured stocks.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Durch die Zunahme von Offshorewindparks wird der anthropogene Lärmpegel im Meer weiter ansteigen. Deshalb ist es notwendig, mögliche negative Effekte von Windturbinenschall auf Fische zu erkennen und zu minimieren. Um mögliche Effekte des Turbinenschalls auf Kabeljau (Gadus morhua) und Scholle (Pleuronectes platessa) zu untersuchen, wurden die Fische in Laborversuchen verschiedenen tiefen Frequenzen und Schallstärken, wie sie von Windturbinen emittiert werden ausgesetzt. Ein ringförmiges Betonversuchsbecken mit 10 Metern Durchmesser wurde mit Schallisolationwänden in verbundene Viertel unterteilt, wodurch im Becken eine Schalldruckdifferenz von 32-52 dB, abhängig von Frequenz und produziertem Schalldruckpegel erzeugt wurde. Die Fische, die sich frei im Becken bewegen und so, wenn sie wollten, den höchsten Schalldruckpegeln ausweichen konnten, wurden monofrequenten Tönen im Frequenzbereich zwischen 25 und 250 Hz in Schalldruckpegeln von 130 und 140 dB re 1 µPa ausgesetzt. Die Position der Fische im Becken wurde in den 24-Stunden-Phasen vor, während und nach Beschallung verglichen und zeigte in den meisten Versuchen während Beschallung signifikant weniger Kabeljau in der Nähe der Schallquelle, als in den Phasen vor und nach Beschallung. Scholle zeigte uneinheitliche Reaktionen, die teilweise auf Meideverhalten, in anderen Versuchen auf Attraktion durch den Schall hindeuteten. Die Ergebnisse lassen wenigstens kurzzeitige, durch Schall verursachte Verteilungsänderungen von Kabeljau in Offshorewindparks erwarten. / The growing number of offshore wind farms will lead to an increase of anthropogenic noise in the sea. Research is therefore necessary to identify and minimize any potentially negative effects of offshore wind turbine noise on fish. To examine the potential impact of offshore wind farm noise emissions on cod (Gadus morhua) and plaice (Pleuronectes platessa), tank experiments were carried out exposing the fishes to different low frequencies at sound levels as emitted by wind turbines An annular concrete tank of 10 m in diameter was divided in connected quarters by sound barriers, so that sound pressure differences of 32 to 52 dB were achieved, depending on frequency and produced sound levels. Pure tones in the frequency range between 25 and 250 Hz were artificially emitted in sound levels of 130 and 140 dB re 1µPa. The experimental fish were free to move around in the tank and therefore could avoid the highest sound levels if they chose to. The location of fish in the 24 hour periods before, during and after sound production were compared and showed significantly lower numbers of cod in the vicinity of the sound source during sound presentation in the majority of the experiments. Reactions of plaice were ambiguous, suggesting both avoidance of, or attraction to the sound. From the results at least short-term changes in distribution of cod related to sound in the vicinity of offshore wind farms could be expected.

Lernaeocera branchialis (Linnaeus, 1767) is a parasitic copepod possessing a complex dual-host lifecycle. The “definitive” gadoid hosts, including Gadus morhua (Atlantic cod), Melanogrammus aeglefinus (haddock) and Merlangius merlangus (whiting), are infected by the fertilised female, which penetrates the host’s ventral aorta or bulbus arteriosus whilst undertaking extensive metamorphosis and a haematophagous lifestyle. The pathogenic effects of this activity upon the host have been well documented and mortality may occur, especially when multiple parasites are present. These negative impacts on cod, particularly juveniles, by L. branchialis have the potential to adversely affect cod aquaculture in the future, and already vulnerable wild cod stocks. This PhD project therefore, investigated the immune response of wild haddock and cultured-cod post-infection by L. branchialis, and the possible mechanisms by which the parasite modulates/evades the host’s immune response. The systemic immune response of both wild haddock and cultured-cod post-infection by L. branchialis depended on the maturation stage of the parasite, and in the former host species, upon the infection intensity. Wild haddock harbouring fully metamorphosed females showed an increase in circulating thrombocytes and a decrease in serum protein levels however; if multiple mature L. branchialis were present the haddock possessed reduced circulating monocytes, and increased circulating thrombocytes and serum anti-trypsin activity. Infection by L. branchialis was also associated with a suppressive effect on haddock serum spontaneous haemolytic activity. These responses were thought to be due to the host trying to counteract the increased damage caused by the massive increase in size and the feeding of the mature parasite, which is more pronounced when multiple parasites are present, resulting in the increase in some parameters and the ‘consumption’ of others. However, the effect of parasite-derived secretions and other pathogens due to observations on wild fish could not be discounted. The laboratory-infection of cultured-cod from two different sources was also performed in order to study the immune response over time. The two groups of cod showed differences in their immune response to L. branchialis. The first group showed suppressed respiratory burst activity of phagocytes, as the parasite reached the early penella sub-stage, whilst no suppression in phagocyte respiratory burst activity was found in the second group. The parasite was found to migrate along the afferent branchial artery of the cod where a thrombus formed and was present throughout its migration into the ventral aorta. At 14 d post-infection, leukocytes expressing Interleukin 8 mRNA were observed within the free-flowing blood at the periphery of the organising thrombus within the lumen of the ventral aorta. This was speculated to aid the recruitment and activation of leukocytes to the site, and the maturation and neovascularisation of granulation tissue. The infection of the second group subsided with the death of the parasite, and none of the parasites metamorphosed past the early penella sub-stage. The live parasites infecting the first group of cod did not possess IgM or complement component C3 binding on their cuticle, however, both IgM and C3 binding occurred on the dead parasites in the second infection trial. This may highlight the importance of these opsonins and the cytotoxic effect of phagocytes in the elimination of L. branchialis by some cod. However, the first infection was terminated as the parasite reached the early penella sub-stage due to a loss of stock cod prior to the study, so the long-term success of the infection can not be concluded. Therefore, the immune response to infection needs to be determined over the entire metamorphosis of L. branchialis to determine whether the infection was successful or not, and preferably in populations with varying susceptibility to L. branchialis. This will not be possible without further studies into the resistance of different stocks of cultured-cod. Many arthropod parasites, such as ticks and salmon lice, have been previously documented to produce pharmacologically active secretions, aiding host invasion and parasite feeding, preventing the host immune response from working effectively against the parasite, all aimed at improving survival of the parasite. Therefore, the effects of the secretory/excretory products (SEPs) produced during the initial infective stage and by the mature, fully metamorphosed female on the immune response of cultured-cod in vitro, and the location of exocrine glands associated with the oral region of the parasite were investigated. The SEPs from the infective stage of the parasite were found not to affect the intracellular hydrogen peroxide (H2O2) production of phagocytes. The practical difficulties in collecting large quantities of the SEPs from the infective stage meant that their effects could not be tested on the other host immune parameters studied. The SEPs from fully metamorphosed female L. branchialis, however, had a number of suppressive effects on the host immune response in vitro including: 1) suppression of the intracellular production of cytotoxic H2O2 during the respiratory burst of phagocytic leukocytes post-PMA stimulation, 2) suppression of the production of macrophage activating factor by leukocytes with a priming effect on naïve phagocyte function, and 3) suppression of the chemo-attraction ‘power’ of zymosan activated cod serum, i.e. anaphylatoxin activity, on head kidney-derived leukocytes. These effects were dose-dependent, and highlight the capacity of L. branchialis to suppress its host’s innate immune response at the local feeding area. Further work is required to establish the mechanisms by which the parasite-derived SEPs suppress these host immune parameters, and to identify which molecules produced by the parasite are responsible. The correlation between these in vitro results, and systemic immune parameters measured from laboratory-infected Atlantic cod and wild infected haddock are discussed. Host immuno-modulation by other arthropod parasites is mediated by pharmacologically active secretions produced by exocrine glands. Therefore, the exocrine glands of the infective and fully metamorphosed female L. branchialis were also investigated in order to identify those that might be responsible for the secretion of host-modifying products. Adult female exocrine glands were mapped using diaminobenzidine (DAB), most commonly known to stain peroxidases and catalases. These compounds are known to be involved in the neutralisation of harmful free radicals which are released during the respiratory burst and tissue damage. Such products may therefore be important protective secretory components at the site of feeding / infection. Exocrine glands were located in the infective stage associated with the oral region, one pair termed the anterior gland complex (AGC), and the other pair extending either side of the oral cone termed the circum-oral glands (CG). These were further investigated using light microscopy and transmission electron microcopy. The AGC and CGs possessed multi-component secretions and they possessed secretory vesicles, abundant and highly active rough endoplasmic reticulum and Golgi apparatus suggesting that protein is an important component of the secretory products. These glands were also observed in the fully metamorphosed females where they had increased in size within the cephalothorax post-metamorphosis. It is hoped that the identification of these glandular structures, which are thought to secrete within the local vicinity of the oral cone, will aid future studies regarding the identification and secretion kinetics of parasite-derived molecules during the infection and feeding process.

<p>Superchilling is a method that can be used to prolong the shelf life of foods by partial freezing. Knowledge of the effects of this method on both the shelf life and quality of foods is important in order to find optimal processing and storage conditions and is of great importance both for the industry and for the consumers. Different raw materials of muscle foods were studied with the purpose of creating a basis for further improvements of both the process and the storage conditions. Products from the commercially important species pork (both roasts and rib steaks), Atlantic salmon (both vacuum packed and wrapped fillets) and Atlantic cod (vacuum packed fillets) were chosen to represent different muscle foods.</p><p>Based on both sensory and microbial evaluations, the superchilled storage improved shelf life of pork roasts from 2 to 16 weeks, and shelf life might even be further prolonged if temperature is kept more constant. The H2S-producing bacteria in superchilled cod fillets did not exceed the limit of consumption during the whole storage period of six weeks, while the microbial shelf life of the ice chilled fillets was estimated to be 8 days after processing.</p><p>Quality changes have been studied with focus on biochemical and physiochemical properties. One of the major goals in meat and fish processing is the ability to retain water and it was observed that drip loss was lower in superchilled samples compared to traditionally chilled samples in all species studied. However, the subsequent liquid loss was higher both in pork roasts and in cod fillets.</p><p>The extractability of protein was used as a tool to monitor protein denaturation. It was found that myofibrillar proteins denatured more easily during superchilled than during traditionally chilled storage both in salmon and in cod fillets. This was confirmed by electrophoresis in salmon. The amount of free amino acids were higher in cod than in salmon and increased more during superchilled storage than during ice chilled storage indicating exoproteolytic activity during storage. Activities of cathepsins B and B + L in salmon fillets were relatively stable during the storage period in all storage groups, demonstrating that these enzymes are not deactivated at the selected storage temperatures and may therefore lead to softening during subsequent chilled storage.</p><p>In superchilled samples of all species studied, white spots emerged on the product surface. However, the appearance of spots did not correspond either to higher drip loss or to higher microbial growth on surface of these samples. On the contrary, the total plate counts of superchilled samples were lower than of the other storage groups. This observation also applied to iron agar counts on cod fillets. These findings are interpreted as a strong indication of that the spots most likely were not of microbial origin.</p><p>The optimal superchilling process and storage conditions remains to be found for the products studied. From the present results it is reasonable to suggest that optimal storage temperature for the vacuum packed salmon fillets is found between the superchilled temperatures in the present experiments. The quality both of the pork roasts and the cod fillets would probably benefit from a storage temperature slightly closer to 0 °C than those studied. It can also be assumed that the control of the superchilling process is more essential to cod than to salmon and pork. In addition, the properties of the raw material are crucial in order to obtain high quality of products after processing and storage.</p> / Paper I-III are reprinted with kind permission from Elsevier, sciencedirect.com

Superchilling is a method that can be used to prolong the shelf life of foods by partial freezing. Knowledge of the effects of this method on both the shelf life and quality of foods is important in order to find optimal processing and storage conditions and is of great importance both for the industry and for the consumers. Different raw materials of muscle foods were studied with the purpose of creating a basis for further improvements of both the process and the storage conditions. Products from the commercially important species pork (both roasts and rib steaks), Atlantic salmon (both vacuum packed and wrapped fillets) and Atlantic cod (vacuum packed fillets) were chosen to represent different muscle foods. Based on both sensory and microbial evaluations, the superchilled storage improved shelf life of pork roasts from 2 to 16 weeks, and shelf life might even be further prolonged if temperature is kept more constant. The H2S-producing bacteria in superchilled cod fillets did not exceed the limit of consumption during the whole storage period of six weeks, while the microbial shelf life of the ice chilled fillets was estimated to be 8 days after processing. Quality changes have been studied with focus on biochemical and physiochemical properties. One of the major goals in meat and fish processing is the ability to retain water and it was observed that drip loss was lower in superchilled samples compared to traditionally chilled samples in all species studied. However, the subsequent liquid loss was higher both in pork roasts and in cod fillets. The extractability of protein was used as a tool to monitor protein denaturation. It was found that myofibrillar proteins denatured more easily during superchilled than during traditionally chilled storage both in salmon and in cod fillets. This was confirmed by electrophoresis in salmon. The amount of free amino acids were higher in cod than in salmon and increased more during superchilled storage than during ice chilled storage indicating exoproteolytic activity during storage. Activities of cathepsins B and B + L in salmon fillets were relatively stable during the storage period in all storage groups, demonstrating that these enzymes are not deactivated at the selected storage temperatures and may therefore lead to softening during subsequent chilled storage. In superchilled samples of all species studied, white spots emerged on the product surface. However, the appearance of spots did not correspond either to higher drip loss or to higher microbial growth on surface of these samples. On the contrary, the total plate counts of superchilled samples were lower than of the other storage groups. This observation also applied to iron agar counts on cod fillets. These findings are interpreted as a strong indication of that the spots most likely were not of microbial origin. The optimal superchilling process and storage conditions remains to be found for the products studied. From the present results it is reasonable to suggest that optimal storage temperature for the vacuum packed salmon fillets is found between the superchilled temperatures in the present experiments. The quality both of the pork roasts and the cod fillets would probably benefit from a storage temperature slightly closer to 0 °C than those studied. It can also be assumed that the control of the superchilling process is more essential to cod than to salmon and pork. In addition, the properties of the raw material are crucial in order to obtain high quality of products after processing and storage. / Paper I-III are reprinted with kind permission from Elsevier, sciencedirect.com