The rocky intertidal zone, experiencing fully marine and fully terrestrial conditions, has become increasingly investigated as a model ecosystem for studying the future implications of climate change. The barnacle, Balanus glandula, a common rocky intertidal inhabitant, plays an important role as a key prey item for many organisms. Low tide can be particularly challenging for barnacles as they are marine organisms subjected to the abiotic conditions of a terrestrial environment. The most stressful of these are increased temperature and decreased oxygen availability. This study aimed to investigate how low tide impacts the energy budget, specifically the digestion, of B. glandula. Barnacles are unable to feed at low tide however, if they were able to digest at low tide, they could maximize their energy intake by emptying their stomach to prepare to feed at the next high tide. However, digestion is a metabolically costly activity, which could make it less energetically favorable to digest when there’s less oxygen available. To test for an effect of low tide on digestion, barnacles were fed, and the time to first fecal production measured as a ‘baseline’. This was repeated, but barnacles were exposed to either a 16ºC or 35 ºC low tide immediately after being fed. The change in digestion time was calculated by comparing these two times for each barnacle. It was found that regardless of temperature, barnacles delayed their digestion by about 50-60 minutes after exposure to a one hour low tide. To determine the energetic cost of digestion, the rate of oxygen consumption was compared between starved and digesting barnacles. I was unable to detect any evidence of elevated metabolic activity during digestion. Additional testing is needed to confirm these results as the barnacles may have not fed during the trial, thus had no food to digest. While it appears that increasing temperatures associated with climate change will have little impact on the digestion of barnacles at low tide, if climate change alters the duration of low tide, there could be an energetic impact to barnacles due to the slowing of their metabolism as indicated by the delay in their digestion.

Like all intertidal species, the barnacle Balanus glandula must cope with temperature and desiccation stress during daily low tide exposure. The increase in temperature at low tide leads to both increased metabolic rate and the potential for increased ATP demand. With its additional inhibition of oxygen intake, low tide thus has an energetic cost that is often reflected in an increase in oxygen consumption following resubmersion. As anthropogenically induced global climate change increases air and water temperatures, its cost might increase. B. glandula individuals were exposed to 4‑hour low tides with maximal temperatures of 18, 30, 35, and 38°C, and their oxygen consumption rates and behaviors were recorded for 4 hours upon resubmersion. It was found that aerial respiration could be measured, though aerial rates were only a fraction of aquatic rates. It was further found that relative aquatic oxygen consumption rates were not elevated following low tide for any temperatures. However, B. glandula individuals exposed to 35 and 38°C low tides remained active a significantly greater portion of time through the first and second hours of recovery, respectively. This indicates that a low tide stress effect is evident in B. glandula, but that it manifests not as an increase in the respiration rate when active, but rather as an increase in the overall activity time. Thus, with increasing global temperatures B. glandula will likely have increased energy needs. This might lead to range relocations, a drive to find new energy sources, and/or reallocations of energy budgets.

As a unique habitat that encompasses steep environmental gradients, it is important to evaluate threats posed to the intertidal zone by rapid climate change. It is thought that intertidal ectotherms are living close to their physiological limit; therefore slight changes in temperature could result in high levels of mortality. Past studies on intertidal species measured thermal tolerance under constant temperatures, neglecting to consider the impacts of natural variation in field temperatures. I conducted a study on the barnacle, Balanus glandula, to assess if a variable thermal environment would alter thermal tolerance. Barnacles were acclimated in an intertidal mesocosm to either daily cold (maximum 20.4◦C), daily warm (maximum 26.5◦C), or variable (two days cold, two days warm) low-tide temperatures. I measured each barnacle’s critical thermal maximum (CTmax) by increasing air temperature 6◦C/hour and identifying the point at which the barnacle ceased to function. Barnacles exposed to any warm temperatures demonstrated an increased thermal tolerance, suggesting that this population of barnacles is capable of shifting their thermal maximum. Furthermore, acclimation to thermal heterogeneity raised thermal maximum, reinforcing the need for future thermal tolerance studies to incorporate biologically-relevant thermal regimes in laboratory experiments. These results demonstrate that B. glandula in the field are well-adapted for increasing air temperatures.

Intertidal habitats are characterized by dynamic, tidally-driven fluctuations in abiotic and biotic factors. Many of the environmental stressors that vary across the intertidal (e.g., temperature, oxygen, food availability, predation pressure) are strong drivers of metabolic rate in ectotherms. As such, we predicted that there may be pronounced differences in the metabolic and stress physiology of conspecific sessile invertebrates occupying at different relative tidal heights. The common acorn barnacle Balanus glandula represents an ideal model organism in which to investigate the possibility of tidal height-dependent physiological differences, owing to their wide distribution in the intertidal zone and their eurytolerant nature. In the first chapter of my thesis, we investigate the hypothesis that B. glandula anchored in the low intertidal have a greater capacity for anaerobic metabolism than conspecifics in the high intertidal, and that this is due to increased predation pressure during submersion. Further, we explore the temporal and spatial fidelity of certain tidal-height driven trends in lactate dehydrogenase activity previously observed in our lab (i.e., higher LDH activity in low intertidal barnacles; Horn et al., 2021), and attempt to identify environmental variables that drive plasticity in LDH activity. We found that, in general, there were higher densities of B. glandula and gastropod whelk predators in the low intertidal compared to the high intertidal, but follow-up studies in the lab revealed that opercular closure in B. glandula was induced by predator exposure (Acanthinucella spirata) for less than 24h. This time frame for shell closure is unlikely to result in internal hypoxia or enhance capacity for anaerobic metabolism. We were therefore not surprised to find that LDH activity in B. glandula was likewise not affected by predator exposures (48h) carried out in the lab. After failing to find an effect of predators on LDH activity in B. glandula, we attempted to replicate the previous finding that LDH activity was highest in low intertidal populations of B. glandula. We did this at the original location in San Luis Obispo Bay, CA as well as at three novel field sites and across seasons and years. While we did observe variation in LDH activity over time and between sites, we did not consistently observe the same trend in LDH activity whereby low intertidal barnacles had the highest activity. In response to these variable patterns, we attempted to identify what environmental parameters, other than predation, might be responsible for plasticity in LDH activity. Unfortunately, neither temperature nor emersion stress – the two variables we examined – had any significant an effect on LDH activity in B. glandula. These data suggest that there must be multiple, interacting stressors – including tidal position - that influence the anaerobic metabolic capacity of B. glandula. In the second chapter of my thesis, we went on to investigate how the response to thermal stress might differ between populations of B. glandula from different vertical heights in the intertidal zone. To this end, we assessed how aerial temperature stress affected oxygen consumption rates (MO2), superoxide dismutase (SOD) activity, and time to mortality in B. glandula collected from both low and high intertidal positions. We found that barnacles from the low intertidal showed a significant increase in MO2 with higher temperature, while MO2 was unaffected by temperature in B. glandula from the high intertidal. We also observed that SOD activity levels were higher in the high intertidal barnacles compared to the low intertidal barnacles, although neither group was increasing SOD activity under higher temperature. Finally, we observed significantly longer survival times during thermal stress in barnacles from the high intertidal zone (e.g., LT50 = 8.75 h vs 5 h at 33˚C for the high and low barnacles, respectively), although this advantage seemed to be lost with the addition of desiccation stress at these same temperatures. It is evident that life in highest reaches of the intertidal zones is physiologically challenging, and this has resulted in a population of B, glandula barnacles that are less sensitive to and better suited to tolerate temperature extremes than conspecifics in the lowest intertidal regions. Understanding how habitat variation may differentially impact the metabolic and thermal stress physiology of B. glandula is increasingly important as climate change progresses. This is particularly significant considering that organisms in the intertidal already reside within a relatively stressful environment and may be living closer to their thermal tolerance limits than animals from less extreme habitats.

The intertidal zone is characterized by persistent, tidally-driven fluctuations in both abiotic (e.g., temperature, [O2], salinity) and biotic (e.g., food availability, predation) conditions, which makes this a very physiologically challenging habitat for resident organisms. The magnitude and degree of variability of these environmental stressors differs between intertidal zones, with the most extreme physiological stress often being experienced by organisms in the high intertidal. Given that many of the fluctuating conditions in this environment are primary drivers of metabolic rate (e.g., temperature, [O2], food availability), we hypothesized that sessile conspecifics residing in different tidal zones would exhibit distinct ‘metabolic phenotypes,’ a term we use to collectively describe the organisms’ baseline metabolic performance and capacity. To investigate this hypothesis, we collected acorn barnacles (Balanus glandula) from low, mid, and high intertidal positions in San Luis Obispo Bay, CA and measured a suite of biochemical (whole-animal citrate synthase (CS) and lactate dehydrogenase (LDH) activity, aerial [lactate]), physiological (O2 consumption rates), morphological (body size), and behavioral (e.g., cirri beat frequency, % time operculum open) indices of metabolism. We found tidal zone-dependent differences in B. glandula metabolism that primarily related to anaerobic capacity, feeding behaviors and body size. Barnacles from the low intertidal tended to have a greater capacity for anaerobic metabolism (i.e., increased LDH activity), feed less when submerged, and be smaller in size compared to conspecifics in the high intertidal. We did not, however, see differences between barnacles from different tidal heights in whole-animal [lactate] following 24h of air exposure, which indicates that the enhanced capacity of low intertidal barnacles for anaerobic metabolism may have evolved to support metabolism during more prolonged episodes of emersion (>>24h) or during events other than emersion (e.g., coastal hypoxia, predation). There were also no significant differences in CS activity or baseline oxygen consumption rates (in air or seawater at 14˚C) across tidal heights, which implies that aerobic metabolic capacity may not be as sensitive to tidal position as anaerobic processes. Understanding how individuals occupying different shore heights differ in their metabolic capacity becomes increasingly interesting in the context of global climate change, given that the intertidal zone is predicted to experience even greater extremes in abiotic stress.