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The purpose of this study was to investigate the effects of hydroxytyrosol (HT) supplementation on markers of mitochondrial biogenesis, aerobic capacity, and endurance exercise performance in recreationally active men. Sixty-one (n = 61) subjects (21.46 ± 0.22 yrs, 179.46 ± 0.79 cm, 78.91 ± 1.19 kg) consumed either a high dose (HI) HT supplement (150 mg HT), a low dose (LO) HT supplement (50 mg HT), or a placebo (PLA) every day for 6 weeks. Muscle biopsies from the vastus lateralis were obtained at baseline and after 6 weeks of supplement consumption and analyzed for markers of mitochondrial biogenesis: succinate dehydrogenase (SDH), citrate synthase (CS), and peroxisome proliferator-activated receptor ɣ coactivator (PGC)-1α. Subjects completed exercise testing on a bicycle ergometer at baseline and after 3 and 6 weeks of supplement consumption to measure changes in maximal aerobic power (VO2MAX), lactate threshold, respiratory exchange ratio (RER), substrate utilization, and endurance exercise performance on a 20 km time trial course. The primary findings were that HT supplementation increased muscle oxidative enzyme activity suggesting increased oxidative capacity. HT also increased time trial performance at midpoint and endpoint and this corresponded with an improvement in lactate threshold and a lower RER for the LO HT treatment. Time trial performance was also improved at endpoint for PLA, however, unlike LO an HI HT, this was accompanied by a significant increase in rating of perceived exercise (RPE) and not associated with improvements in muscle oxidative capacity. Our results indicate that HT ranging from 50 to 150 mg/day for 6 weeks can improve muscle oxidative capacity and aerobic performance, and suggests that HT may be used chronically to improve mitochondrial function. HT may be used as an effective means to increase mitochondria to improve exercise performance, and limit diseases associated with mitochondrial dysfunction such as cardiovascular disease, type II diabetes, and some cancers. / text

This thesis is organized in “sandwich” format, as recommended by my supervisory committee. It consists of three main chapters. Chapter one is a general introduction and outlines the background information leading to the objectives and hypotheses of my thesis research. Chapter two is a manuscript prepared for submission to a peer-reviewed scientific journal. Chapter three is an overview of the major findings of this thesis, their implications in fish physiology and ecotoxicology, including suggestions of future directions of research. Appendix A contains data from an additional series of experiments that were conducted during my thesis but are not included as a full data chapter. It will be prepared for publication after my defence. / Hypoxia often occurs in aquatic ecosystems that receive effluent from municipal wastewater treatment plants (WWTP). WWTP effluent contains contaminants that could disrupt the complex physiological pathways fish use to cope with hypoxia (e.g., pharmaceuticals, polychlorinated biphenyls, and polycyclic aromatic hydrocarbons), but the effects of WWTP effluent on the physiological responses of fish to chronic hypoxia is poorly understood. We exposed mummichog killifish (Fundulus heteroclitus) to hypoxia (5 and 2 kPa O2) and/or WWTP effluent for 21 days in a full factorial design. We then measured hypoxia tolerance, whole-animal metabolism, gill morphology, haematology, and tissue metabolites. In clean water, killifish responded to chronic hypoxia with improvements in hypoxia tolerance – increases in time to loss of equilibrium at 0.5 kPa (tLOE) and decreases in critical O2 tension (Pcrit) – in association with increased gill surface area as a result of regression of the interlamellar cell mass (ILCM). Concurrent exposure to wastewater attenuated the increases in tLOE and gill remodeling in chronic hypoxia, and nearly depleted brain glycogen stores. Therefore, exposure to WWTP effluent can disrupt the physiological mechanisms fish use to cope with chronic hypoxia and impair hypoxia tolerance. My research suggests that the combination of stressors near WWTPs can have interactive effects on the physiology and health of fish. / Thesis / Master of Science (MSc) / Low oxygen conditions, known as ‘hypoxia’, frequently occur in aquatic ecosystems that receive municipal wastewater treatment plant (WWTP) effluent. WWTP effluent is a continuous and complex source of pollution, including contaminants that can disrupt fish physiology, affecting their ability to cope with stressors, like hypoxia. The effects of WWTP effluent on the responses of fish to chronic hypoxia are poorly understood. To address this research gap, I examined the effects of hypoxia and WWTP effluent on chronically exposed mummichog killifish. I provide evidence that combined exposure to hypoxia and wastewater affected hypoxia tolerance, gill structure, and depleted energy stores in the brain. My thesis demonstrates that WWTP effluent can disrupt mechanisms that fish use to cope with chronic hypoxia and impair hypoxia tolerance. These findings contribute to the existing body of work that documents the interactive effects of combined stressors in effluent-dominated ecosystems on the physiology and health of fish.

Salinity and oxygen availability have long been recognised as important factors influencing animal physiology and therefore species distribution. The maintenance of appropriate cellular ion levels is critical for many essential physiological processes, but at the same time is energetically expensive. Since hypoxia is likely to impose aerobic limitations for ATP generation, the maintenance of salt and water homeostasis could be at risk during hypoxia. The amphidromous inanga (Galaxias maculatus) is well known for its salinity tolerance and its life cycle that involves several salinity related migrations. During these migrations inanga also frequently encounters hypoxic waters, and therefore must maintain energy homeostasis when aerobic metabolism may be compromised. The present study has investigated behavioural, physiological, biochemical and molecular mechanisms by which inanga tolerate changes in salinity and hypoxia. After 14 days of acclimation to salinities ranging from freshwater to 43‰, inanga showed physiological acclimation. This was evident by no changes in metabolic rates or energy expenditures through this salinity range. Energy balance seemed to be tightly and efficiently controlled by changes in the proportion of protein and lipids used as energy substrate. No mortalities and only minor changes in plasma osmolality also indicated salinity acclimation. The remarkable osmoregulatory capacity of inanga was also evidenced after a seawater challenge. The osmotic balance of inanga was only disrupted during the first 24 hours after the challenge, evidenced by an increase in plasma osmolality and plasma Na+, and a decrease in muscle water content. These physiological changes were correlated with changes at the molecular level. Different isoforms of the catalytic subunit of the Na+,K+-ATPase (NKA) were isolated, partially sequenced and identified in inanga. Phylogenetic analysis grouped inanga isoforms (α-1a, α-1b, α-1c) with their respective homologues from salmonids. Patterns of mRNA expression were also similar to salmonids, with α-1a being downregulated and α-1b being up-regulated following seawater challenge. Previous to this study, NKA isoform switching was reported to occur only in salmonids and cichlids. The presence of NKA subunits that change with environmnetal salinity in inanga indicates that this isoform switching phenomenon is much more widespread among teleost lineages than previously thought. Aiming to elucidate the hypoxia tolerance of inanga, oxygen consumption rate as a function of decreasing external PO2 was evaluated. At no point did inanga regulate oxygen consumption, suggesting that this species is an oxyconformer. This is the first robust demonstration of the existence of oxyconforming in fish. Evaluation of the scaling relationship between oxygen consumption and fish size in normoxia, showed that the exponent of this relationship fell within the range previously reported for fish. However, in hypoxic conditions the scaling relationship was less clear suggesting different size-related mechanisms for tolerating hypoxia. Analysis of the aerobic and anaerobic metabolism of small and large fish, showed that smaller inanga were able to sustain aerobic metabolism for longer than larger inanga, which instead relied on anaerobic metabolism for extending their survival. This knowledge is likely to be of value for the conservation of this iconic fish species, by incorporating these size related differences in hypoxia tolerance in streams management. In light of the unusual oxyconforming response of inanga, a study examining the behavioural responses of this species to declining dissolved oxygen was performed. Inanga did not display a behaviour that might reduce energy expenditure during oxygen limitation; instead swimming activity and speed were elevated relative to normoxia. As hypoxia deepened inanga leaped out of the water, emersing themselves on a floating platform. Once emersed, fish exhibited an enhanced oxygen consumption rate compared to fish that remained in hypoxic water. Although this emersion behaviour was hypothesised to be of physiological advantage, both aquatic hypoxia and emersion resulted in similar physiological and biochemical consequences in inanga. While in hypoxic water oxygen availability seemed to be the limiting factor, in air failure of the circulatory system was hypothesised to be the cause of a similar metabolic signature to that found in aquatic hypoxia. Overall, inanga seemed to be not particularly well adapted to tolerate aquatic hypoxia. In light of the increasing likelihood of anthropogenic-induced hypoxia in inanga habitats, this is likely to have negative consequences for the future of inanga populations in the wild. Although this study provides the mechanisms behind the exceptional salinity tolerance of inanga, its susceptibility to hypoxia is likely to impose further constraints for the osmoregulatory processes that guarantee inanga survival during life cycle migrations. The results of the present study are relevant for understanding and managing the fishery of this economically- and culturally important fish species.

La zone côtière Atlantique constitue une importante nourricerie pour les juvéniles de sole commune (Solea solea, Linnaeus 1758). Pour évaluer les conséquences sur la biologie des juvéniles de cette espèce exploitant un milieu côtier de plus en plus anthropisé, les effets de l’hypoxie et/ou des Polychlorobiphényles (PCBs) ont été évalués sur leurs performances physiologiques (métabolisme aérobie et performances cardiaques) ainsi que sur leur comportement vis-à-vis des prédateurs. Pour cela, une étude a été réalisée à deux échelles d’organisation biologique, l’organe et l’individu. Des contaminations aux PCBs (670 ng and 2239 ng PCB g-1 de nourriture ont été effectuées expérimentalement par voie trophique pendant 30 et 60 jours. De telles expositions ont engendré des effets peu marqués sur le métabolisme aérobie, se traduisant par une faible augmentation du métabolisme de maintenance associé à une augmentation de la concentration critique en oxygène (O2crit) chez les soles exposées sur le long terme à la plus haute concentration de PCB. De la même façon, la performance cardiaque n’est apparue que très peu affectée par la contamination aux PCBs. Nous avons tout de même noté une diminution de la force de contraction du cœur des soles contaminées lorsqu’il est fortement sollicité comme c’est le cas au cours d’une activité physique soutenue. Par ailleurs, le mécanisme de contraction du cœur se trouve modifié chez les soles contaminées avec une participation plus importante du calcium issu du réticulum sarcoplasmique que chez les soles témoins. Les résultats concernant le comportement anti-prédateur ont montré que l’hypoxie engendrait une réduction du taux de réponse de fuite de la sole, ainsi qu’une limitation de leur capacité cryptique via une augmentation oxy-dépendent de la fréquence de ventilation. D’une façon générale, même si les PCBs semblent avoir peu d’effets sur les performances physiologiques testées, il ne peut être exclu qu’ils puissent compromettre à plus long terme le développement et la survie des juvéniles de soles, qu’ils soient associés ou non à des conditions environnementales limitantes comme l’hypoxie. / The Atlantic coastal zone constitutes an important nursery for the juveniles of the common sole (Solea solea, Linnaeus on 1758). To assess the effect of anthropisation on the survival of the juveniles of common sole, the effects of the hypoxia and\or Polychlorinated Biphenyles (PCBs) on their physiological performances (aerobic metabolism and cardiac performances) were assessed as well as their antipredator behaviour. For that purpose, the study was carried out at two levels of biological organization, the organ and the individual. Sole contamination with PCB (670 ng and 2239 ng PCB g-1 of food), was made experimentally via the trophic pathway, during 30 and 60 days. Our results suggest that PCBs slightly affected the aerobic metabolism. An increase of the standard metabolic rate participate to raise the level of critical oxygen concentration (CritO2) in soles exposed to long term to the highest concentration of PCB was noted. In the same way, the cardiac performances seem to be slightly affected by PCB-contamination. A decrease in the peak tension was noted in PCB-contaminated soles forced to an intense activity. In addition it was noted an increase in the participation of the sarcoplasmic reticulum to the force of contraction in the hearts of soles contaminated compared with hearts of control soles. The results obtained with regard to the simulation of the attack of a predator showed that responsiveness was lower in hypoxia compared with normoxia associated with a decrease of their cryptic capacity. In general, even if the PCB seems to have slight effects on the tested physiological performances, it cannot be excluded that long term exposure to PCBs can compromise the development and the survival of the juveniles of common sole, independently of their association with limiting environmental conditions such as hypoxia.

Rodd är en sport som kräver hög kondition då de flesta av kroppens stora muskler används och kroppens alla tre energisystem är inblandade. Därmed har kroppen ett stort behov av energi vid rodd.    Roddmaskin är en utrustning som används av roddare under vintersäsongen för att utveckla prestationen och göra fysiologiska tester.    Syftet med projektet var att undersöka hur takt (drag per minut) och styrka (1RM marklyft) påverkade, sluttiden (s) vid 1000 m rodd på roddergometrar. Studien utfördes på 9 manliga deltagare över 18 år och sambanden studerades med linjär regressionsanalys. Hypotesen var att lägre takt leder till högre kraft i draget och högre takt leder till lägre kraft. Resultaten visade inget samband mellan takt, högre vikt i marklyft och sluttid där en lägre takt och högre medeleffekt gav en lägre sluttid, p och r2 värdet för sambandet mellan tid och takt var 0.109 för r2och p=0.385, p= 0.210 och r2 värdet 0.214 för sambandet mellan tid och marklyft, för sambandet mellan tid och medeleffekt var r2 värdet 0.75 och p = 0.0025, vilket tyder på ett starkt samband.    Det är troligen större fördel om man drar med större kraft på roddmaskin än om man skulle ro på vatten och resultaten är därför inte direkt överförbara till rodd på vatten. Vidare antyder resultaten att det kan vara en fördel om roddarna tränar mer styrka då mer styrka leder till kraftigare drag, vilket i sin tur leder till en lägre tid. / Rowing is a sport dependent on high endurance capacity, where most of the large muscles in the body are used, the three energy systems are involved and the body is in need of a large amount of energy. Concept 2 rowing ergometers is a sport of itself but is performed mostly during the winter season for outdoor rowers to see the performance and do physiological tests. The purpose of this project was to see if there is any correlation between stroke rate, average power, 1RM deadlift and finishing time on 1000 m rowing machine performance, for 9 adult male rowers. The correlation was studied with linear regression analysis.    The hypothesis was that lower rate leads to higher mean power in each stroke and higher stroke rate leads to lower power in each stroke.    The results showed no correlation between stroke rate, heavier weight lifted in deadlift and finishing time. Where a lower stroke rate and higher mean effect lead to a lower finishing time, p and r2 value for the correlation between time and stroke rate was 0.109 for r2 and p=0.385, p=0.210 and r2 value 0.214 for the correlation between time and deadlift, for the correlation between time and mean effect, the r2 value was 0.75 and p=0.0025, which indicate a strong correlation.        It is more benefit if one row with higher force on the rowing machine than rowing on the water. Further, the results show that it is an advantage if the rowers train more strength, because more power leads to a more powerful stroke and lower time.

Introduction : Les muscles paravertébraux interviennent en permanence dans les tâches de la vie quotidienne pour la stabilisation et la mobilisation du rachis. Cela nécessite la contribution du métabolisme aérobie musculaire. La lombalgie chronique est caractérisée par un haut niveau de fatigabilité et des sensations de douleur au regard des muscles paravertébraux, qui pourraient être en partie attribués à l’altération des réponses du métabolisme aérobie musculaire, tandis que la pratique d’activités physiques et sportives (APS) pourrait améliorer ces réponses Objectif : L’objectif général était de déterminer la spécificité des réponses du métabolisme aérobie des muscles paravertébraux chez des patients lombalgiques chroniques, avant et après un programme de restauration fonctionnelle, et chez différents athlètes en bonne santé. Méthode : Quatre études ont été menées. Chaque participant a effectué des exercices isocinétiques sous-maximaux d’extension du tronc pendant cinq minutes, afin d’évaluer les réponses du métabolisme aérobie des muscles paravertébraux à l’exercice. Des sujets lombalgiques chroniques ont été appariés et comparés à des sujets sains, puis suivis au cours d’un programme de restauration fonctionnelle, dont la principale composante était l’exercice physique. Des sujets jeunes et en bonne santé, sportifs et non sportifs, ont été comparés entre eux. Résultats/Conclusion : Chez les lombalgiques chroniques, le programme de restauration fonctionnel a permis une accélération des cinétiques de V̇O2, une augmentation de l’oxygénation musculaire, et une augmentation de l’efficience mécanique à l’exercice. Chez les jeunes sportifs, les réponses aérobies étaient aussi améliorées, et la modification de ces réponses était dépendante des modalités d’entrainement. Ces résultats suggèrent que la pratique d’APS améliore la contribution du métabolisme aérobie au cours d’un exercice prolongé d’extension du tronc, au regard des muscles paravertébraux. Il n’y avait pas de différence entre les réponses métaboliques et hémodynamiques à l’exercice des lombalgiques chroniques, et celles des sujets sains, malgré un faible niveau de force maximale, et un haut niveau de fatigabilité musculaire à l’exercice chez les lombalgiques chroniques. Bien que les résultats préliminaires ne témoignent pas d’une altération des réponses chez les lombalgiques chroniques, nos travaux suggèrent que la pratique d’activités physiques pourrait diminuer le niveau de fatigabilité des muscles paravertébraux, et ainsi limiter les risques de blessure et de contre-performance, en améliorant les réponses aérobies à l’exercice. / Introduction: Paraspinal muscles are required continuously during daily tasks for trunk stabilization and mobilization. For this, aerobic metabolism muscle contribution is required. Chronic low back pain is characterized by a high level of fatigability and pain sensations in regard to paraspinal muscles. These symptoms may be attributed to an alteration in aerobic metabolism responses. Conversely, practising physical and sports activities (PSA) could improve these responses.Objective: The main objective was to determine the specificities in aerobic metabolism responses in regard to paraspinal muscles, in chronic low back pain patients following a functional restoration program and in healthy athletes.Method: Four studies have been conducted. Participants performed submaximal trunk extension exercises on an isokinetic dynamometer for five minutes, to assess paraspinal muscle aerobic metabolism responses to exercises. Chronic low back pain patients were paired with and compared to healthy individuals and followed during a functional restoration program, in which physical exercise was the main component. Also, healthy young individuals, with and without sport specialization, were compared with each other.Results/Conclusion: In chronic low back pain patients, the functional restoration program allows an acceleration in V̇O2, kinetics, an increase in paraspinal muscle oxygenation, and an increase in mechanical efficiency to exercise. In young athletes, the aerobic responses were also improved, and the changes in responses depended on the training modalities. Practicing SPA enhances the aerobic metabolism contribution during prolonged trunk extension exercise, in regard to paraspinal muscles. There was no difference in aerobic metabolism responses between the chronic low back pain patients and the healthy individuals, despite weak levels of maximal strength and high levels of muscle fatigability when exercising (for the chronic low back pain patients). Although the preliminary results did not attest to an alteration in aerobic responses in chronic low back pain patients, our results suggest that practising physical activities could limit paraspinal muscle fatigability, which could be a way to prevent injury and disappointing performances, by improving aerobic metabolism responses to exercise.