The ecological impacts of multiple environmental stressors on coastal biofilm bacteria.

Ecological communities are increasingly exposed to multiple interacting stressors. For example, warming directly affects the physiology of organisms, eutrophication stimulates the base of the food web, and harvesting larger organisms for human consumption dampens top-down control. These stressors often combine in the natural environment with unpredictable results. Bacterial communities in coastal ecosystems underpin marine food webs and provide many important ecosystem services (e.g. nutrient cycling and carbon fixation). Yet, how microbial communities will respond to a changing climate remains uncertain. Thus, we used marine mesocosms to examine the impacts of warming, nutrient enrichment, and altered top-predator population size structure (common shore crab) on coastal microbial biofilm communities in a crossed experimental design. Warming increased bacterial α-diversity (18% increase in species richness and 67% increase in evenness), but this was countered by a decrease in α-diversity with nutrient enrichment (14% and 21% decrease for species richness and evenness, respectively). Thus, we show some effects of these stressors could cancel each other out under climate change scenarios. Warming and top-predator population size structure both affected bacterial biofilm community composition, with warming increasing the abundance of bacteria capable of increased mineralization of dissolved and particulate organic matter, such as Flavobacteriia, Sphingobacteriia, and Cytophagia. However, the community shifts observed with warming depended on top-predator population size structure, with Sphingobacteriia increasing with smaller crabs and Cytophagia increasing with larger crabs. These changes could alter the balance between mineralization and carbon sequestration in coastal ecosystems, leading to a positive feedback loop between warming and CO2 production. Our results highlight the potential for warming to disrupt microbial communities and biogeochemical cycling in coastal ecosystems, and the importance of studying these effects in combination with other environmental stressors.

Amelioration of ocean acidification and warming effects through physiological buffering of a macroalgae.

Concurrent anthropogenic global climate change and ocean acidification are expected to have a negative impact on calcifying marine organisms. While knowledge of biological responses of organisms to oceanic stress has emerged from single-species experiments, these do not capture ecologically relevant scenarios where the potential for multi-organism physiological interactions is assessed. Marine algae provide an interesting case study, as their photosynthetic activity elevates pH in the surrounding microenvironment, potentially buffering more acidic conditions for associated epiphytes. We present findings that indicate increased tolerance of an important epiphytic foraminifera, Marginopora vertebralis, to the effects of increased temperature (±3°C) and pCO2 (~1,000 µatm) when associated with its common algal host, Laurencia intricata. Specimens of M. vertebralis were incubated for 15 days in flow-through aquaria simulating current and end-of-century temperature and pH conditions. Physiological measures of growth (change in wet weight), calcification (measured change in total alkalinity in closed bottles), photochemical efficiency (Fv/Fm), total chlorophyll, photosynthesis (oxygen flux), and respiration were determined. When incubated in isolation, M. vertebralis exhibited reduced growth in end-of-century projections of ocean acidification conditions, while calcification rates were lowest in the high-temperature, low-pH treatment. Interestingly, association with L. intricata ameliorated these stress effects with the growth and calcification rates of M. vertebralis being similar to those observed in ambient conditions. Total chlorophyll levels in M. vertebralis decreased when in association with L. intricata, while maximum photochemical efficiency increased in ambient conditions. Net production estimates remained similar between M. vertebralis in isolation and in association with L. intricata, although both production and respiration rates of M. vertebralis were significantly higher when associated with L. intricata. These results indicate that the association with L. intricata increases the resilience of M. vertebralis to climate change stress, providing one of the first examples of physiological buffering by a marine alga that can ameliorate the negative effects of changing ocean conditions.

Seahorse Hotels: Use of artificial habitats to support populations of the endangered White's seahorse Hippocampus whitei.

The provision of temporary, specially designed artificial habitat may help support populations of the Endangered Whites' seahorse Hippocampus whitei in the face of rapid coastal urbanisation and declining natural habitats. Three designs of artificial habitat (Seahorse Hotels) were installed in Port Stephens, New South Wales, Australia, where natural habitats had significantly declined. Mark recapture surveys were used to assess seahorse site fidelity and population parameters, and the effect of Seahorse Hotel design on seahorse abundance, epibiotic growth and mobile epifaunal seahorse prey was determined. The Seahorse Hotels sustained a substantial population of seahorses (64; 57-72 95% confidence intervals) in comparison to recent local population estimates. There were no significant differences in seahorse abundance, mobile epifauna or epibiotic growth among the three different hotel designs. This research demonstrated that H. whitei will inhabit Seahorse Hotels in absence of natural habitat, and additional complexity in these artificial structures was not necessary to support seahorse populations. Temporary structures such as Seahorse Hotels will be a valuable tool in supporting H. whitei and other Syngnathid populations through infrastructure maintenance or habitat modification.

The endangered White's seahorse Hippocampus whitei chooses artificial over natural habitats.

To explore whether the endangered White's seahorse Hippocampus whitei would choose to inhabit artificial over natural habitats, 10 adult H. whitei individuals were put through a series of binary choice trials in aquaria, during which they were offered different paired combinations of natural (different types of macroalga and seagrass) and artificial habitat (panels of swimming-net material). It was found that H. whitei displayed a significant choice for swimming-net material over all other available natural habitats and chose habitats according to the following rankings: (a) Net; (b) Sargassum sp.; (c) Posidonia australis; (d) Zostera muelleri. Hippocampus whitei's choice of swimming net material over natural habitat suggests that these artificial structures could be a useful conservation measure for seahorses in areas where natural habitat is becoming less favourable due to declines in abundance or quality.

Can transplanting enhance mobile marine invertebrates in ecologically engineered rock pools?

The field of eco-engineering has burgeoned in recent years in response to the proliferation of artificial structures. Adding water-retaining features to seawalls has been successful in increasing biodiversity relative to the surrounding structure. Artificial rock pools may not, however, completely mimic natural rock pools. Here, we compared natural colonisation, through dispersal and recruitment, of intertidal mobile species to water-retaining flowerpots on seawalls with that into rock pools. This represents the more usual 'passive' approach to eco-engineering where features are built to enhance biodiversity and are allowed to colonise naturally, as opposed to seeding or transplanting organisms to features. While flowerpots supported some mobile species not found on the seawall, other species common on natural shores did not recruit to flowerpots. Thus, in a second experiment we tested the effectiveness of an 'active' approach through transplanting mobile organisms to flowerpots to expedite the colonisation process. For the species examined, however, most individuals did not stay in the flowerpots for more than 24 h after being transplanted. Further understanding of the processes (e.g. dispersal distances, recruitment) influencing colonisation of eco-engineered habitats is needed to effectively inform management of marine infrastructure, particularly for projects targeted at restoration rather than enhancement.

When time affects space: Dispersal ability and extreme weather events determine metacommunity organization in marine sediments.

Community ecology has traditionally assumed that the distribution of species is mainly influenced by environmental processes. There is, however, growing evidence that environmental (habitat characteristics and biotic interactions) and spatial processes (factors that affect a local assemblage regardless of environmental conditions - typically related to dispersal and movement of species) interactively shape biological assemblages. A metacommunity, which is a set of local assemblages connected by dispersal of individuals, is spatial in nature and can be used as a straightforward approach for investigating the interactive and independent effects of both environmental and spatial processes. Here, we examined (i) how environmental and spatial processes affect the metacommunity organization of marine macroinvertebrates inhabiting the intertidal sediments of a biodiverse coastal ecosystem; (ii) whether the influence of these processes is constant through time or is affected by extreme weather events (storms); and (iii) whether the relative importance of these processes depends on the dispersal abilities of organisms. We found that macrobenthic assemblages are influenced by each of environmental and spatial variables; however, spatial processes exerted a stronger role. We also found that this influence changes through time and is modified by storms. Moreover, we observed that the influence of environmental and spatial processes varies according to the dispersal capabilities of organisms. More effective dispersers (i.e., species with planktonic larvae) are more affected by spatial processes whereas environmental variables had a stronger effect on weaker dispersers (i.e. species with low motility in larval and adult stages). These findings highlight that accounting for spatial processes and differences in species life histories is essential to improve our understanding of species distribution and coexistence patterns in intertidal soft-sediments. Furthermore, it shows that storms modify the structure of coastal assemblages. Given that the influence of spatial and environmental processes is not consistent through time, it is of utmost importance that future studies replicate sampling over different periods so the influence of temporal and stochastic factors on macrobenthic metacommunities can be better understood.

Ocean acidification but not warming alters sex determination in the Sydney rock oyster, Saccostrea glomerata.

Whether sex determination of marine organisms can be altered by ocean acidification and warming during this century remains a significant, unanswered question. Here, we show that exposure of the protandric hermaphrodite oyster, Saccostrea glomerata to ocean acidification, but not warming, alters sex determination resulting in changes in sex ratios. After just one reproductive cycle there were 16% more females than males. The rate of gametogenesis, gonad area, fecundity, shell length, extracellular pH and survival decreased in response to ocean acidification. Warming as a sole stressor slightly increased the rate of gametogenesis, gonad area and fecundity, but this increase was masked by the impact of ocean acidification at a level predicted for this century. Alterations to sex determination, sex ratios and reproductive capacity will have flow on effects to reduce larval supply and population size of oysters and potentially other marine organisms.

Increasing habitat complexity on seawalls: Investigating large- and small-scale effects on fish assemblages.

The construction of artificial structures in the marine environment is increasing globally. Eco-engineering aims to mitigate the negative ecological impacts of built infrastructure through designing structures to be multifunctional, benefiting both humans and nature. To date, the focus of eco-engineering has largely been on benefits for benthic invertebrates and algae. Here, the potential effect of eco-engineered habitats designed for benthic species on fish was investigated. Eco-engineered habitats ("flowerpots") were added to an intertidal seawall in Sydney Harbour, Australia. Responses of fish assemblages to the added habitats were quantified at two spatial scales; large (among seawalls) and small (within a seawall). Data were collected during high tide using cameras attached to the seawall to observe pelagic and benthic fish. At the larger spatial scale, herbivores, planktivores, and invertebrate predators were generally more abundant at the seawall with the added flowerpots, although results were temporally variable. At the smaller spatial scale, certain benthic species were more abundant around flowerpots than at the adjacent control areas of seawall, although there was no general pattern of differences in species density and trophic group abundance of pelagic fish between areas of the seawall with or without added habitats. Although we did not find consistent, statistically significant findings throughout our study, the field of research to improve fish habitat within human-use constraints is promising and important, although it is in its early stages (it is experimental and requires a lot of trial and error). To advance this field, it is important to document when effects were detected, and when they were not, so that others can refine the designs or scale of habitat enhancements or their study approaches (e.g., sampling protocols).

Effect of copper on multiple successional stages of a marine fouling assemblage.

Copper based paints are used to prevent fouling on the hulls of ships. The widely documented effect of copper on hull assemblages may be primarily due to direct effects on the invertebrates themselves or indirect effects from copper absorbed into the microbial biofilm before settlement has commenced. Artificial units of habitat were exposed to varied regimes of copper to examine (1) the photosynthetic efficiency and pigments of early-colonising biofilms, and (2) subsequent macroinvertebrate assemblage change in response to the different regimes of copper. Macroinvertebrate assemblages were found to be less sensitive to the direct effects of copper than indirect effects as delivered through biofilms that have been historically exposed to copper, with some species more tolerant than others. This raises further concern for the efficacy of copper as a universal antifoulant on the hulls of ships, which may continue to assist the invasion of copper-tolerant invertebrate species.

Ocean acidification narrows the acute thermal and salinity tolerance of the Sydney rock oyster Saccostrea glomerata.

Coastal and estuarine environments are characterised by acute changes in temperature and salinity. Organisms living within these environments are adapted to withstand such changes, yet near-future ocean acidification (OA) may challenge their physiological capacity to respond. We tested the impact of CO2-induced OA on the acute thermal and salinity tolerance, energy metabolism and acid-base regulation capacity of the oyster Saccostrea glomerata. Adult S. glomerata were acclimated to three CO2 levels (ambient 380µatm, moderate 856µatm, high 1500µatm) for 5weeks (24°C, salinity 34.6) before being exposed to a series of acute temperature (15-33°C) and salinity (34.2-20) treatments. Oysters acclimated to elevated CO2 showed a significant metabolic depression and extracellular acidosis with acute exposure to elevated temperature and reduced salinity, especially at the highest CO2 of 1500µatm. Our results suggest that the acute thermal and salinity tolerance of S. glomerata and thus its distribution will reduce as OA continues to worsen.

Storm effects on intertidal invertebrates: increased beta diversity of few individuals and species.

Climate change is predicted to lead to more extreme weather events, including changes to storm frequency, intensity and location. Yet the ecological responses to storms are incompletely understood for sandy shorelines, the globe's longest land-ocean interface. Here we document how storms of different magnitude impacted the invertebrate assemblages on a tidal flat in Brazil. We specifically tested the relationships between wave energy and spatial heterogeneity, both for habitat properties (habitat heterogeneity) and fauna (ß-diversity), predicting that larger storms redistribute sediments and hence lead to spatially less variable faunal assemblages. The sediment matrix tended to become less heterogeneous across the flat after high-energy wave events, whereas ß-diversity increased after storms. This higher ß-diversity was primarily driven by species losses. Significantly fewer species at a significantly lower density occurred within days to weeks after storms. Negative density and biomass responses to storm events were most prominent in crustaceans. Invertebrate assemblages appeared to recover within a short time (weeks to months) after storms, highlighting that most species typical of sedimentary shorelines are, to some degree, resilient to short-term changes in wave energy. Given that storm frequency and intensity are predicted to change in the coming decades, identifying properties that determine resilience and recovery of ecosystems constitute a research priority for sedimentary shorelines and beyond.

Incorporating in situ habitat patchiness in site selection models reveals that site fidelity is not always a consequence of animal choice.

Understanding site fidelity is important in animal ecology, but evidence is lacking that this behaviour is due to an animal choosing a specific location. To discern site selection behaviour, it is necessary to consider the spatial distribution of habitats that animals can occupy within a landscape. Tracking animals and defining clear habitat boundaries, however, is often difficult. We use in situ habitat distribution data and animal movement simulations to investigate behavioural choice in site fidelity patterns. We resolved the difficulty of gathering data by working with intertidal rock pool systems, which are of manageable size and where boundaries are easy to define. Movements of the intertidal starfish Parvulastra exigua were quantified to test the hypotheses that (1) this species displays fidelity to a particular rock pool and that (2) rock pool fidelity is due to site selection behaviour. Observed patterns of individuals (n = 10 starfish) returning to a previously occupied rock pool (n = 5 pools per location) were tested against an expected null distribution generated through simulations of random movements within their natural patchy environment. Starfish exhibited site selection behaviour at only one location even though site fidelity was high (av. 7·4 starfish out of 10 found in test pools) in two of the three locations. The random chance of a starfish returning to a pool increased 67% for each metre further a rock pool was from the original pool, and 120% for each square metre increase in surface area of an original pool. The decision of returning to an original rock pool was influenced by food availability. When microalgal cover was >60%, there was a c. 50% chance of animals staying faithful to that pool. Our results show the importance to consider spatial distribution of habitats in understanding patterns of animal movement associated with animal choices and site fidelity. Returning to a particular place does not necessarily mean that an animal is homing; it may be the only place to go.

Adult exposure to ocean acidification is maladaptive for larvae of the Sydney rock oyster Saccostrea glomerata in the presence of multiple stressors.

Parental effects passed from adults to their offspring have been identified as a source of rapid acclimation that may allow marine populations to persist as our surface oceans continue to decrease in pH. Little is known, however, whether parental effects are beneficial for offspring in the presence of multiple stressors. We exposed adults of the oyster Saccostrea glomerata to elevated CO2 and examined the impacts of elevated CO2 (control = 392; 856 µatm) combined with elevated temperature (control = 24; 28°C), reduced salinity (control = 35; 25) and reduced food concentration (control = full; half diet) on their larvae. Adult exposure to elevated CO2 had a positive impact on larvae reared at elevated CO2 as a sole stressor, which were 8% larger and developed faster at elevated CO2 compared with larvae from adults exposed to ambient CO2 These larvae, however, had significantly reduced survival in all multistressor treatments. This was particularly evident for larvae reared at elevated CO2 combined with elevated temperature or reduced food concentration, with no larvae surviving in some treatment combinations. Larvae from CO2-exposed adults had a higher standard metabolic rate. Our results provide evidence that parental exposure to ocean acidification may be maladaptive when larvae experience multiple stressors.

A limited legacy effect of copper in marine biofilms.

The effects of confounding by temporal factors remains understudied in pollution ecology. For example, there is little understanding of how disturbance history affects the development of assemblages. To begin addressing this gap in knowledge, marine biofilms were subjected to temporally-variable regimes of copper exposure and depuration. It was expected that the physical and biological structure of the biofilms would vary in response to copper regime. Biofilms were examined by inductively coupled plasma optical emission spectrometry, chlorophyll-a fluorescence and field spectrometry and it was found that (1) concentrations of copper were higher in those biofilms exposed to copper, (2) concentrations of copper remain high in biofilms after the source of copper is removed, and (3) exposure to and depuration from copper might have comparable effects on the photosynthetic microbial assemblages in biofilms. The persistence of copper in biofilms after depuration reinforces the need for consideration of temporal factors in ecology.

The interplay between habitat structure and chemical contaminants on biotic responses of benthic organisms.

Habitat structure influences the diversity and distribution of organisms, potentially affecting their response to disturbances by either affecting their 'susceptibility' or through the provision of resources that can mitigate impacts of disturbances. Chemical disturbances due to contamination are associated with decreases in diversity and functioning of systems and are also likely to increase due to coastal urbanisation. Understanding how habitat structure interacts with contaminants is essential to predict and therefore manage such effects, minimising their consequences to marine systems. Here, we manipulated two structurally different habitats and exposed them to different types of contaminants. The effects of contamination and habitat structure interacted, affecting species richness. More complex experimental habitats were colonized by a greater diversity of organisms than the less complex habitats. These differences disappeared, however, when habitats were exposed to contaminants, suggesting that contaminants can override effects of habitats structure at small spatial scales. These results provide insight into the complex ways that habitat structure and contamination interact and the need to incorporate evidence of biotic responses from individual disturbances to multiple stressors. Such effects need to be taken into account when designing and planning management and conservation strategies to natural systems.

Facing the Heat: Does Desiccation and Thermal Stress Explain Patterns of Orientation in an Intertidal Invertebrate?

A key challenge for ecologists is to quantify, explain and predict the ecology and behaviour of animals from knowledge of their basic physiology. Compared to our knowledge of many other types of distribution and behaviour, and how these are linked to individual function, we have a poor level of understanding of the causal basis for orientation behaviours. Most explanations for patterns of animal orientation assume that animals will modify their exposure to environmental factors by altering their orientation. We used a keystone grazer on rocky shores, the limpet Cellana tramoserica, to test this idea. Manipulative experiments were done to evaluate whether orientation during emersion affected limpet desiccation or body temperature. Body temperature was determined from infrared thermography, a technique that minimises disturbance to the test organism. No causal relationships were found between orientation and (i) level of desiccation and (ii) their body temperature. These results add to the growing knowledge that responses to desiccation and thermal stress may be less important in modifying the behaviour of intertidal organisms than previously supposed and that thermoregulation does not always reflect patterns of animal orientation. Much of what we understand about orientation comes from studies of animals able to modify orientation over very short time scales. Our data suggests that for animals whose location is less flexible, orientation decisions may have less to do with responses to environmental factors and more to do with structural habitat properties or intrinsic individual attributes. Therefore we suggest future studies into processes affecting orientation must include organisms with differing levels of behavioural plasticity.

Size-balanced community reorganization in response to nutrients and warming.

It is widely accepted that global warming will adversely affect ecological communities. As ecosystems are simultaneously exposed to other anthropogenic influences, it is important to address the effects of climate change in the context of many stressors. Nutrient enrichment might offset some of the energy demands that warming can exert on organisms by stimulating growth at the base of the food web. It is important to know whether indirect effects of warming will be as ecologically significant as direct physiological effects. Declining body size is increasingly viewed as a universal response to warming, with the potential to alter trophic interactions. To address these issues, we used an outdoor array of marine mesocosms to examine the impacts of warming, nutrient enrichment and altered top-predator body size on a community comprised of the predator (shore crab Carcinus maenas), various grazing detritivores (amphipods) and algal resources. Warming increased mortality rates of crabs, but had no effect on their moulting rates. Nutrient enrichment and warming had near diametrically opposed effects on the assemblage, confirming that the ecological effects of these two stressors can cancel each other out. This suggests that nutrient-enriched systems might act as an energy refuge to populations of species under metabolic constraints due to warming. While there was a strong difference in assemblages between mesocosms containing crabs compared to mesocosms without crabs, decreasing crab size had no detectable effect on the amphipod or algal assemblages. This suggests that in allometrically balanced communities, the expected long-term effect of warming (declining body size) is not of similar ecological consequence to the direct physiological effects of warming, at least not over the six week duration of the experiment described here. More research is needed to determine the long-term effects of declining body size on the bioenergetic balance of natural communities.

Patterns of the non-indigenous isopod Cirolana harfordi in Sydney Harbour.

Biological introductions can alter the ecology of local assemblages and are an important driver of global environmental change. The first step towards understanding the impact of a non-indigenous species is to study its distribution and associations in the invaded area. In Sydney Harbour, the non-indigenous isopod Cirolana harfordi has been reported in densities up to 0.5 individuals per cm(2) in mussel-beds. Abundances of this species have, however, been largely overlooked in other key habitats. The first aim of this study was to evaluate the abundances and distribution of C. harfordi across different habitats representative of Sydney Harbour. Results showed that C. harfordi occurred in oyster and mussel-beds, being particularly abundant in oyster-beds. We also aimed to determine the role of C. harfordi as a predator, scavenger and detritus feeder by investigating the relationships between densities of C. harfordi and (i) the structure of the resident assemblages, and (ii) deposited organic matter in oyster-beds. Densities of C. harfordi were not related to the structure of the assemblages, nor amounts of deposited organic matter. These findings suggested little or no ecological impacts of C. harfordi in oyster-beds. These relationships may, however, affect other variables such as growth of individuals, or be disguised by high variability of assemblages among different locations. Future studies should, therefore, test the impacts of C. harfordi on the size of organisms in the assemblage and use manipulative experiments to control for spatial variation. This study is the first published work on the ecology of the invasion of C. harfordi and provides the starting-point for the study of the impacts of this species in Sydney Harbour.

Trying to fit in: are patterns of orientation of a keystone grazer set by behavioural responses to ecosystem engineers or wave action?

The distribution of animals varies at different temporal and spatial scales. At the smallest scale, distribution may be orientated in regard to particular environmental variables or habitat features. For animals on the rocky intertidal, the processes which set and maintain patterns of distribution and abundance in wave-exposed areas are well studied, with explanatory models focused on wave action and, more recently, the role of biogenic habitats. In contrast, patterns of orientation by intertidal animals have received less attention, although having ecological and fitness consequences. Here, we report tests of competing models to explain the observation that limpets on steeply sloped surfaces orientate downwards. A greater proportion of downwards-facing limpets was found in sheltered sites and areas without barnacles and this pattern was consistent across many shores and sampling occasions. Additionally, the frequency at which limpets were dislodged after a storm was independent of orientation. To test whether orientation is a behavioural response to habitat-forming barnacles, barnacles were removed and/or killed from patches of substrata and the change in proportion of downwards-facing limpets measured. The proportion increased with barnacle removal and this behaviour was a response to the structure of the barnacles, not a biotic effect associated with the living organism. Our study suggests that biogenic habitat not wave action sets patterns of limpet orientation and barnacle shells, regardless of whether the barnacle is alive or not, limit the ability of limpets to adopt a downward orientation.

Effect of extraction-method, period of incubation and tidal emersion on the viability of haemocytes from oysters.

The impacts of pollution on marine organisms are often investigated using the viability of their haemocytes. Although this assay is routinely used in monitoring, field and laboratory experimentation, there has been less effort in further optimizing procedures to reduce artefacts and facilitate sampling over large geographic areas. Using the oyster Saccostrea glomerata as a model species, we investigated the effects of different techniques for extracting haemolymph, period of incubation with dye and emersion-time (e.g. tidal-state) on the viability of haemocytes. Collecting haemocytes with a syringe, through a drilled hole in the shell, increased the viability of haemocytes by almost 50%. While emersion-time and incubating haemocytes with the dye for up to 4 h did not affect viability. This simple in situ approach provides a less destructive method for extracting haemocytes, allowing their viability to be measured as part of large-scale experiments without jeopardizing the surrounding assemblage of animals and plants.