The studies presented in this thesis were designed to investigate the effects of reduced sea water pH on the behaviour of intertidal decapod crustaceans, both within the context of the variations occurring naturally in the pH of rock pool habitats, and in relation to predicted changes to ocean pH resulting from ocean acidification and potential carbon dioxide (CO2) leaks from carbon capture storage (CCS) sites. Recent studies on marine fish have shown behavioural disruptions as a result of increased CO2 concentrations in sea water and reduced pH, but the effects on crustaceans are as yet unknown. The first two studies investigated the effects of reduced pH upon the olfactory behaviour of the prawn Palaemon elegans and the hermit crab Pagurus bernhardus, focussing on their responses to food odours. Short-term (five day) exposures to highly reduced pH (pHNBS = 6.60, 6.80) revealed disruptions to the chemo-sensory behaviour of both species with a reduction in their ‘sniffing’ response, and the inability of P. bernhardus to locate the chemical cue. This was also accompanied by elevated haemolymph chloride ions. in In a further study P. bernhardus was subjected to a longer exposure (60 days) and to a range of pH levels (pHNBS = 8.00, 7.90, 7.70, 7.35 and 6.80) in order to detect a threshold for the behavioural disruptions observed, and to determine if there would be any sign of acclimation over a longer period. A clear gradient in the disruptions to the chemo-sensory responses and survival rates of the hermit crabs, and disruption to a physiological marker (elevated haemolymph calcium ions), was found. Possible thresholds for disruption were also identified at levels that match predictions for ocean acidification and leaks from proposed CO2 CCS sites. Some of the crabs in the lower pH treatments exhibited a recovery in their responses by day 60, possibly indicating an acclimation effect. The presence of disruption to haemolymph ion concentrations in both the short and longer term hermit crab studies suggest a mechanism for behavioural disruption. In a final study the effects of reduced sea water pH on a more complex behaviour, involving decision making, was investigated. Reduced sea water pH was shown to disrupt the shell assessment and selection behaviour of P. bernhardus affecting its decision making processes, although not all crabs were affected in the same way. The work presented here therefore demonstrates that reduced sea water pH could have disruptive effects upon both information gathering, via chemo-sensory processes, and decision making in intertidal crustaceans. The mechanism responsible is unlikely to be due to changes in the odour molecule, or physical damage to receptor organs. Rather the observed disruptions could be due (a) to ionic changes, causing metabolic depression or interference with neurotransmitter function, or (b) to disruption to chemoreception per se. Such disturbances to key behavioural processes have implications for inter and intraspecific species interactions and population dynamics in the marine environment. Changes in pH are already experienced by intertidal animals for short periods when rock pools are emersed, but future anthropogencially-induced reductions in sea water pH are likely to cause more sustained and widespread disruptions with, as yet, unpredictable consequences. The differential responses observed between individuals in these studies may warrant further investigation as such differences may provide the basis for selection and adaptation to projected changes in ocean pH.

Wastewater disposal often has deleterious impacts on the receiving environment. Low levels of dissolved oxygen are of particular concern. Here I investigate the impacts on dissolved oxygen and carbon chemistry of screened municipal wastewater in the marine waters off Victoria, B.C., Canada. I analyzed data from a series of undersea moorings, ship-based monitoring, and underwater remotely-operated vehicle video. I used these observations to construct a two-layer box model of the nearfield receiving environment. Despite the lack of more advanced treatment, dissolved oxygen levels near the outfalls are well above the commonly used 63 umol kg-1 hypoxic threshold and that the outfalls’ impact on water column oxygen is likely less than a few umol kg-1. Likewise, dissolved inorganic carbon is not elevated and pH not depressed compared to the surrounding region. Strong tidal currents and the cold, well-oxygenated waters of Victoria’s marine environment give these waters a high assimilative capacity for organic waste. / Graduate / 0595 / 0403 / 0543 / jeremykrogh@gmail.com

Since the 1800s, carbon dioxide emissions due to human activities have contributed significantly to the amount of carbon in the atmosphere. Approximately a third of this carbon is absorbed by the ocean, through air-sea fluxes at the ocean surface (Sabine, 2004). Increased CO2 has changed the carbon chemistry of the ocean and hence the pH. pH is expected to drop by 0.4 by the year 2100. It is unclear how this lower pH will affect carbon cycling and sequestration with respect to the biological carbon pump. Most studies have focused on open ocean phytoplankton or bacterial communities in large, stationary mesocosms. Few studies have coupled both phytoplankton and bacterial processes and even fewer have investigated coastal communities, where pH and pCO2 can vary drastically. This study focused first on developing and evaluating a mesocosm and alternative method for elevating pCO2. The second goal was to determine how potential changes in phytoplankton DOC release and community structure and the resulting carbon pool may affect bacterial secondary production and ectoenzyme activity in a natural coastal community. Mesocosms aimed to mimic natural pCO2 fluctuations by maintaining CO2 concentration of 1250 ppm in the headspace, as aqueous pCO2 may change with biological processes. Six mesocosms were filled with 40L of water from the Chesapeake Bay (three ambient pCO2 and three 1250 ppm) and monitored over 15 days. Chlorophyll a, DOC, bacterial respiration, bacterial production, and enzyme activity were measured. Bacterial production and respiration were used to calculate bacterial growth efficiency (BGE). Results showed that there was no significant difference between the ambient and elevated groups with respect to chlorophyll a, DOC, BGE and enzymes activity. However, differences in bacterial respiration and bacterial production during the first four days of the experiment may suggest that bacteria require time to acclimate to elevated pCO2. Phytoplankton and bacteria in coastal areas are exposed to a wide range of abiotic factors such as seasonal temperature variations, salinity, mixing, and terrestrial inputs. The pH of the Chesapeake Bay ranges between 7.5 and 8.3, and it is possible that the phytoplankton and bacteria are adapted to cope with a wide range of pH (Wong, 2012). This study suggests that the biological carbon pump may not be significantly altered in our future ocean.

Anthropogenic activities are fundamentally altering the chemistry of the world’s oceans. Many of these modifications could have a significant impact on the health of marine organisms. Yet, despite being proposed as one of the most significant threats that marine ecosystems face, to date very little is known about the impact of anthropogenic climate change, and ocean acidification in particular, on host defence. The aims of this thesis are to investigate the impact of environmental stressors on the invertebrate immune response, providing empirical data on how anthropogenically induced stressors will impact the invertebrate immune system and how this will impact organism condition and subsequent physiological trade-offs. Exposure to reduced seawater pH and increased temperature significantly reduced the immune response in the blue mussel, Mytilus edulis. This reduction in immune response could indicate stress-induced immune dysfunction. However, the immune system protects an organism from infectious disease, ensuring survival, and should therefore be evaluated functionally rather than immunologically. By subsequently exposing mussels to a bacterial challenge this study demonstrated that an earlier study which measured a reduction in host defence represented a trade-off of immune system maintenance costs, with mussels maintaining a capacity to up-regulate immune defence when required. However, whilst this immune plasticity ensures mussels are able to survive a pathogen exposure, such a strategy appears to be physiologically costly. This cost is seen as a reduction in reproductive investment, an altered energy metabolism and an altered fatty acid composition in organisms exposed to low pH. Therefore the overarching picture that emerges is, without measuring physiological processes functionally, and in neglecting any physiological trade-offs, it is possible that many studies may misinterpret the complex physiological responses of marine organisms to ocean acidification.

Le cycle océanique de l'azote est à l'origine de deux rétro-actions climatiques au sein du système terre. D'unepart, il participe au contrôle du réservoir d'azote fixé disponible au développement du phytoplancton et à lamodulation de la pompe biologique, un des mécanismes de séquestration du carbone anthropique. D'autre part,le cycle de l'azote produit un gaz à effet de serre et destructuer d'ozone, le protoxyde d'azote (N2O). L'évolutionfuture du cycle de l'azote sous l'influence du rechauffement climatique, de la déoxygénation et de l'acidificationdes océans reste une question ouverte. Les processus tels que la fixation d'azote, la dénitrification et laproduction de protoxyde d'azote seront modifiés sous l'influence conjuguée des ces trois stresseurs. Cesinteractions peuvent être évaluées grâce aux modèles globaux de biogéochimie marine. Nous utilisons NEMOPISCESet l'ensemble des modèles CMIP5 pour projeter les modifications des taux de fixation d'azote, denitrification, de production et des flux air-mer de N2O à l'horizon de 2100 en réponse au scénario 'business-asusual'.Les effets liés à l'action combinée du rechauffement climatique et de l'acidification des océans sur leréservoir d'azote fixé, la production primaire et la rétro-action sur le bilan radiatif sont également évalués danscette thèse.! / The marine nitrogen cycle is responsible for two climate feedbacks in the Earth System. Firstly, it modulates thefixed nitrogen pool available for phytoplankton growth and hence it modulates in part the strength of thebiological pump, one of the mechanisms contributing to the oceanic uptake of anthropogenic CO2. Secondly, thenitrogen cycle produces a powerful greenhouse gas and ozone (O3) depletion agent called nitrous oxide (N2O).Future changes of the nitrogen cycle in response to global warming, ocean deoxygenation and oceanacidification are largely unknown. Processes such as N2-fixation, nitrification, denitrification and N2Oproduction will experience changes under the simultaneous effect of these three stressors. Global oceanbiogeochemical models allow us to study such interactions. Using NEMO-PISCES and the CMIP5 modelensemble we project changes in year 2100 under the business-as-usual high CO2 emissions scenario in globalscale N2-fixation rates, nitrification rates, N2O production and N2O sea-to-air fluxes adding CO2 sensitivefunctions into the model parameterizations. Second order effects due to the combination of global warming intandem with ocean acidification on the fixed nitrogen pool, primary productivity and N2O radiative forcingfeedbacks are also evaluated in this thesis.

Thesis: Ph. D., Joint Program in Biological Oceanography (Massachusetts Institute of Technology, Department of Biology; and the Woods Hole Oceanographic Institution), 2017. / Cataloged from PDF version of thesis. / Includes bibliographical references (pages 159-168). / Thecosome pteropods are planktonic mollusks that form aragonite shells and that may experience increased dissolution and other adverse effects due to ocean acidification. This thesis focuses on assessing the possible biological effects of ocean acidification on the shells and locomotion of pteropods and examining the response of a local pteropod population to environmental change over time. I analyzed shell condition after exposing pteropods to elevated CO₂ as well as in natural populations to investigate the sensitivity of the shells of different species to aragonite saturation state ([omega][subscript A]). The pteropods (Limacina retroversa) from laboratory experiments showed the clearest pattern of shell dissolution in response to decreased [omega][subscript A], while wild populations either had non-significant regional trends in shell condition (Clio pyramidata) or variability in shell condition that did not match expectations due to regional variability in [omega][subscript A] (Limacina helicina). At locations with intermediate [omega][subscript A] (1.5-2.5) the variability seen in L. helicina shell condition might be affected by food availability more than tA. I examined sinking and swimming behaviors in the laboratory in order to investigate a possible fitness effect of ocean acidification on pteropods. The sinking rates of L. retroversa from elevated CO₂ treatments were slower in conjunction with worsened shell condition. These changes could increase their vulnerability to predators in the wild. Swimming ability was mostly unchanged by elevated CO₂ after experiments that were up to three weeks in duration. I used a long-term dataset of pteropods in the Gulf of Maine to directly test whether there has been a population effect of environmental change over the past several decades. I did not observe a population decline between 1977 and 2015, and L. retroversa abundance in the fall actually increased over the time series. Analysis of the habitat use of L. retroversa revealed seasonal associations with temperature, salinity, and bottom depths. The combination of laboratory experiments and field surveys helped to address gaps in knowledge about pteropod ecology and improve our understanding of the effects of ocean acidification on pteropods. / by Alexander Bergan. / Ph. D.

Diante do quadro atual de previsões de mudanças climáticas, estudos a respeito das possíveis respostas dos organismos a estas alterações são importantes. Com a finalidade de prever e verificar se estas serão de fato deletérias ou se os organismos são capazes de lidar com elas sem alterações na sua fisiologia, e consequentemente na estrutura do ambiente, E. brasiliensis foi utilizada como modelo para estudar possíveis impactos do aumento da temperatura e acidificação dos oceanos na sua fisiologia. Para isso, espécimes foram expostos a 9 possíveis combinações de temperatura (24ºC, 28ºC e 30ºC) e pH (8.0, 7.7 e 7.3) em diferentes intervalos de tempo (1, 3, 12, 24 e 48 h). Amostras de gônadas e fluido celomático foram coletadas para avaliar a expressão das proteínas de estresse HSP70, AIF-1 e p38-MAPK, e a variação no número e viabilidade dos celomócitos. Nossos resultados mostram que o modelo é sensibilizado pelas mudanças no ambiente, através da hiper-regulação das proteínas de estresse. O cenário considerado mais extremo (30°C + pH7.3) ocasionou a morte de 100% dos organismos após 24horas. E o segundo cenário mais severo (30°C + pH7.7) desencadeou o desenvolvimento de ulceração de pele. Os efeitos são mais pronunciados nos celomócitos e a acidificação da água parece ter efeitos antagônicos com a temperatura nos celomócitos e sinérgicos nas gônadas. Embora a resposta tenha sido sistêmica, o grau e a dinâmica foram distintos em relação às diferentes amostras e estresses. Podendo causar modificações na resposta imune dos organismos e consequentemente na sobrevivência da espécie a longo prazo. / Under the current Climate Change context, studies about the potential responses of the organisms to their changing environment are of extreme importance. Recent studies point out the synergy of temperature and ocean acidification altogether. In this study, we used the sea star E. brasiliensis to assess the physiological effects of rising temperature, seawater acidification and the interaction of both factors. Independent individuals (N=225) were exposed to 9 possible combinations of temperature (24ºC, 28ºC and 30ºC) and pH (8.0, 7.7 and 7.3), for 1, 3, 12, 24 and 48 h. We compared the stress produced by these treatments measuring the expression of heat shock proteins (HSP70), the production of the allograft inflammatory factor (AIF−1) and the activation of mitogen kinases (MAPKs) at both gonad and celomic fluid. Furthermore, we assessed the quantity and quality of coelomocytes. Our results demonstrated that E. brasiliensis is vulnerable to the interaction of temperature and acidification. All the stress proteins evaluated were upregulated. The extreme scenario (30°C + pH7.3) caused the death of 100% of organisms after 24 hours, while the second most severe scenario (30°C + pH7.7) triggered skin ulceration. Nevertheless, we found that water acidification produces antagonistic effects to the temperature in coelomocytes and synergistic effects in gonad cells. Furthermore, these effects were more pronounced in the coelomocytes than in the gonads. The systemic response found in this study suggest that the interactive effects of elevated temperatures in conjunction with ocean acidification may endanger the survival of this species, and it could compromise the ecosystem functioning at long term.

World-wide coral reefs are in a state of decline as a result of many local and regional factors. Recent global mass mortalities of reef corals due to record warm sea temperatures have led researchers to consider global warming as one of the most significant threats to the persistence of coral reef ecosystems over the next 100 years. It is well established that elevated sea surface temperatures (SSTs) cause widespread coral bleaching, yet confusion lingers as to what facet of extreme temperatures is most important. Utilizing long-term in situ datasets, nine thermal stress indices were calculated and their effectiveness at segregating bleaching years a posteriori for multiple reefs on the Florida Reef Tract was tested. Simple bleaching thresholds based on deviations above the climatological maximum monthly SST were just as effective at identifying bleaching years as complex thermal stress indices. Near real-time bleaching alerts issued by NOAA's Integrated Coral Observing Network (ICON) are now based upon a running 30-day average SST, such that alerts are only issued when the running 30-day average SST exceeds the estimated bleaching threshold for a particular site. In addition to three widespread, mass-coral bleaching events, the Florida Reef Tract was impacted by three tropical storms and 12 hurricanes from 1997-2005. Sea surface cooling associated with the high frequency of hurricanes that impacted Florida in 2005 likely acted to ameliorate the severity and duration of bleaching. Nonetheless, hurricane-associated cooling is not expected to nullify the proposed effects of climate change on coral reefs. The role of thermal stress in coral bleaching has been extensively studied for eastern tropical Pacific (ETP) coral reefs. However, the ETP presents a unique opportunity as this region has sub-optimal conditions for coral reef development because of upwelling of carbon dioxide-enriched deep water along the shallow thermocline. This upwelling results in a depressed aragonite saturation state which is likely an additional factor in the poor reef development throughout the ETP. The highest aragonite saturation documented in this study occur in the Gulf of Chiriquí, which corresponds with the greatest reef development of the entire ETP. Seasonal upwelling had a significant effect on the carbonate chemistry of surface waters in Pacific Panamá. This regionally-depressed aragonite saturation state of the ETP appears to result in corals with a less dense skeleton. Density values of poritid corals from the Galápagos, where aragonite saturation was the lowest documented in this study, were significantly less dense relative to those from Panamá and the Great Barrier Reef. The density of non-living pocilloporid framework components were no different across the ETP aragonite saturation gradients. This could be a result of the activity of boring sponges removing the primary carbonate material within the dead coral skeleton, thus lowering density, albeit physical-chemical dissolution cannot be ruled out. These studies provide real-world examples of the ramifications of global climate change upon coral reef ecosystems.

Animals experience variation in their environment because of natural changes. However, due to anthropogenic disturbance, the speed and severity of these changes have recently increased. This thesis investigates how reproductive behaviours may be affected by human induced environmental change. In specific, I investigate how visual and chemical changes in the aquatic environment, caused by eutrophication, affect mating systems and sexual selection in fish. Broad-nosed- and straight-nosed pipefish, which both have been studied in detail for a long period, were used as model organisms. These two species are particularly suitable model organisms since they perform complex courtship behaviours, including the advertisement of ornaments and a nuptial dance. Further, two distinct populations were studied, one on the Swedish west coast and one in the Baltic Sea, as these two locations vary in the degree and extent of environmental disturbance, in particular turbidity. I found that changes in the visual environment had no impact on the development of female sexual ornaments in these sex-role reversed pipefishes, but it hampered adaptive mate choice. Turbidity also had a negative effect on reproductive success in the Baltic Sea population. Changes in the chemical environment in the form of increased pH reduced the probability to mate, while hypoxia did not alter mating propensity. However, hypoxic water delayed the onset of both courting and mating. Hence, human induced change in aquatic environments may alter the processes of sexual selection and population dynamics.

As human activities continue to generate accelerating levels of carbon dioxide emissions, the world’s oceanic resources are threatened by variability in seawater chemistry, known as ocean acidification. Recent increases in atmospheric carbon dioxide have resulted in decreased carbonate ion concentrations and ocean pH levels, leading to increasingly acidic waters. The exact consequences of these chemical changes on ecosystems and individual species are difficult to predict; however, research has shown that economically valuable calcifying species will experience reduced reproductive fitness and population declines. Ocean acidification, therefore, poses an immediate risk to both fish stocks and fishery industries. From a local perspective, individual regions will need to implement dynamic management strategies to prepare for anticipated economic consequences. In a global context, international cooperation is required for further research and collaborative efforts must be made to reduce future acidification.