From transcriptome to biological function: environmental stress in an ectothermic vertebrate, the coral reef fish Pomacentrus moluccensis

Background
Our understanding of the importance of transcriptional regulation for biological function is continuously improving. We still know, however, comparatively little about how environmentally induced stress affects gene expression in vertebrates, and the consistency of transcriptional stress responses to different types of environmental stress. In this study, we used a multi-stressor approach to identify components of a common stress response as well as components unique to different types of environmental stress. We exposed individuals of the coral reef fish Pomacentrus moluccensis to hypoxic, hyposmotic, cold and heat shock and measured the responses of approximately 16,000 genes in liver. We also compared winter and summer responses to heat shock to examine the capacity for such responses to vary with acclimation to different ambient temperatures.
Results
We identified a series of gene functions that were involved in all stress responses examined here, suggesting some common effects of stress on biological function. These common responses were achieved by the regulation of largely independent sets of genes; the responses of individual genes varied greatly across different stress types. In response to heat exposure over five days, a total of 324 gene loci were differentially expressed. Many heat-responsive genes had functions associated with protein turnover, metabolism, and the response to oxidative stress. We were also able to identify groups of co-regulated genes, the genes within which shared similar functions.
Conclusion
This is the first environmental genomic study to measure gene regulation in response to different environmental stressors in a natural population of a warm-adapted ectothermic vertebrate. We have shown that different types of environmental stress induce expression changes in genes with similar gene functions, but that the responses of individual genes vary between stress types. The functions of heat-responsive genes suggest that prolonged heat exposure leads to oxidative stress and protein damage, a challenge of the immune system, and the re-allocation of energy sources. This study hence offers insight into the effects of environmental stress on biological function and sheds light on the expected sensitivity of coral reef fishes to elevated temperatures in the future.

Heterologous microarray experiments used to identify the early gene response to heat stress in a coral reef fish

Coral reef fishes are expected to experience rising sea surface temperatures due to climate change. How well tropical reef fishes will respond to these increased temperatures and which genes are important in the response to elevated temperatures is not known. Microarray technology provides a powerful tool for gene discovery studies, but the development of microarrays for individual species can be expensive and time-consuming. In this study, we tested the suitability of a Danio rerio oligonucleotide microarray for application in a species with few genomic resources, the coral reef fish Pomacentrus moluccensis. Results from a comparative genomic hybridization experiment and direct sequence comparisons indicate that for most genes there is considerable sequence similarity between the two species, suggesting that the D. rerio array is useful for genomic studies of P. moluccensis. We employed this heterologous microarray approach to characterize the early transcriptional response to heat stress in P. moluccensis. A total of 111 gene loci, many of which are involved in protein processing, transcription, and cell growth, showed significant changes in transcript abundance following exposure to elevated temperatures. Changes in transcript abundance were validated for a selection of candidate genes using quantitative real-time polymerase chain reaction. This study demonstrates that heterologous microarrays can be successfully employed to study species for which specific microarrays have not yet been developed, and so have the potential to greatly enhance the utility of microarray technology to the field of environmental and functional genomics.

Small within-day increases in temperature affects boldness and alters personality in coral reef fish

Consistent individual differences in behaviour, termed personality, are common in animal populations and can constrain their responses to ecological and environmental variation, such as temperature. Here, we show for the first time that normal within-daytime fluctuations in temperature of less than 3°C have large effects on personality for two species of juvenile coral reef fish in both observational and manipulative experiments. On average, individual scores on three personality traits (PTs), activity, boldness and aggressiveness, increased from 2.5- to sixfold as a function of temperature. However, whereas most individuals became more active, aggressive and bold across temperature contexts (were plastic), others did not; this changed the individual rank order across temperatures and thus altered personality. In addition, correlations between PTs were consistent across temperature contexts, e.g. fish that were active at a given temperature also tended to be both bold and aggressive. These results (i) highlight the importance of very carefully controlling for temperature when studying behavioural variation among and within individuals and (ii) suggest that individual differences in energy metabolism may contribute to animal personality, given that temperature has large direct effects on metabolic rates in ectotherms.

Using otolith microchemistry and shape to assess the habitat value of oil structures for reef fish

Over 7500 oil and gas structures (e.g. oil platforms) are installed in offshore waters worldwide and many will require decommissioning within the next two decades. The decision to remove such structures or turn them into reefs (i.e. 'rigs-to-reefs') hinges on the habitat value they provide, yet this can rarely be determined because the residency of mobile species is difficult to establish. Here, we test a novel solution to this problem for reef fishes; the use of otolith (earstone) properties to identify oil structures of residence. We compare the otolith microchemistry and otolith shape of a site-attached coral reef fish (Pseudanthias rubrizonatus) among four oil structures (depth 82-135 m, separated by 9.7-84.2 km) on Australia's North West Shelf to determine if populations developed distinct otolith properties during their residency. Microchemical signatures obtained from the otolith edge using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) differed among oil structures, driven by elements Sr, Ba and Mn, and to a lesser extent Mg and Fe. A combination of microchemical data from the otolith edge and elliptical Fourier (shape) descriptors allowed allocation of individuals to their 'home' structure with moderate accuracy (overall allocation accuracy: 63.3%, range: 45.5-78.1%), despite lower allocation accuracies for each otolith property in isolation (microchemistry: 47.5%, otolith shape: 45%). Site-specific microchemical signatures were also stable enough through time to distinguish populations during 3 separate time periods, suggesting that residence histories could be recreated by targeting previous growth zones in the otolith. Our results indicate that reef fish can develop unique otolith properties during their residency on oil structures which may be useful for assessing the habitat value of individual structures. The approach outlined here may also be useful for determining the residency of reef fish on artificial reefs, which would assist productivity assessments of these habitats.

Application of species distribution models to explain and predict the distribution, abundance and assemblage structure of nearshore temperate reef fishes

 Aim: The purpose of this study was to create predictive species distribution models (SDMs) for temperate reef-associated fish species densities and fish assemblage diversity and richness to aid in marine conservation and spatial planning. Location: California, USA. Methods: Using generalized additive models, we associated fish species densities and assemblage characteristics with seafloor structure, giant kelp biomass and wave climate and used these associations to predict the distribution and assemblage structure across the study area. We tested the accuracy of these predicted extrapolations using an independent data set. The SDMs were also used to estimate larger scale abundances to compare with other estimates of species abundance (uniform density extrapolation over rocky reef and density extrapolations taking into account variations in geomorphic structure). Results: The SDMs successfully modelled the species-habitat relationships of seven rocky reef-associated fish species and showed that species' densities differed in their relationships with environmental variables. The predictive accuracy of the SDMs ranged from 0.26 to 0.60 (Pearson's r correlation between observed and predicted density values). The SDMs created for the fish assemblage-level variables had higher prediction accuracies with Pearson's r values of 0.61 for diversity and 0.71 for richness. The comparisons of the different methods for extrapolating species densities over a single marine protected area varied greatly in their abundance estimates with the uniform extrapolation (density values extrapolated evenly over the rocky reef) always estimating much greater abundances. The other two methods, which took into account variation in the geomorphic structure of the reef, provided much lower abundance estimates. Main conclusions: Species distribution models that combine geomorphic, oceanographic and biogenic habitat variables can reliably predict spatial patterns of species density and assemblage attributes of temperate reef fishes at spatial scales of 50 m. Thus, SDMs show great promise for informing spatial and ecosystem-based approaches to conservation and fisheries management. © 2015 John Wiley

Using community-based monitoring with GIS to create habitat maps for a marine protected area in Australia

In recent years there has been an increase in community-based monitoring programmes developed and implemented worldwide. This paper describes how the data collected from such a programme could be integrated into a Geographic Information System (GIS) to create temperate subtidal marine habitat maps. A differential Global Positioning System was utilized to accurately record the location of the trained community-based SCUBA diver data. These georeferenced data sets were then used to classify benthic habitats using an aerial photograph and digitizing techniques. This study demonstrated that trained community-based volunteers can collect data that can be utilized within a GIS to create reliable and cost-effective maps of shallow temperate subtidal rocky reef systems.

An evaluation of Sea Search as a citizen science programme in Marine Protected Areas

Citizen science involves collaboration between multi-sector agencies and the public to address a natural resource management issue. The Sea Search citizen science programme involves community groups in monitoring and collecting subtidal rocky reef and intertidal rocky shore data in Victorian Marine Protected Areas (MPAs), Australia. In this study we compared volunteer and scientifically collected data and the volunteer motivation for participation in the Sea Search programme. Intertidal rocky shore volunteer-collected data was found to be typically comparable to data collected by scientists for species richness and diversity measures. For subtidal monitoring there was also no significant difference for species richness recorded by scientists and volunteers. However, low statistical power suggest only large changes could be detected due to reduced data replication. Generally volunteers recorded lower species diversity for biological groups compared to scientists, albeit not significant. Species abundance measures for algae species were significantly different between volunteers and scientists. These results suggest difficulty in identification and abundance measurements by volunteers and the need for additional training requirements necessary for surveying algae assemblages. The subtidal monitoring results also highlight the difficulties of collecting data in exposed rocky reef habitats with weather conditions and volunteer diver availability constraining sampling effort. The prime motivation for volunteer participation in Sea Search was to assist with scientific research followed closely by wanting to work close to nature. This study revealed two important themes for volunteer engagement in Sea Search: 1) volunteer training and participation and, 2) usability of volunteer collected data for MPA managers. Volunteer-collected data through the Sea Search citizen science programme has the potential to provide useable data to assist in informed management practices of Victoria’s MPAs, but requires the support and commitment from all partners involved.

Assessment of habitat representation across a network of marine protected areas with implications for the spatial design of monitoring

Networks of marine protected areas (MPAs) are being adopted globally to protect ecosystems and supplement fisheries management. The state of California recently implemented a coast-wide network of MPAs, a statewide seafloor mapping program, and ecological characterizations of species and ecosystems targeted for protection by the network. The main goals of this study were to use these data to evaluate how well seafloor features, as proxies for habitats, are represented and replicated across an MPA network and how well ecological surveys representatively sampled fish habitats inside MPAs and adjacent reference sites. Seafloor data were classified into broad substrate categories (rock and sediment) and finer scale geomorphic classifications standard to marine classification schemes using surface analyses (slope, ruggedness, etc.) done on the digital elevation model derived from multibeam bathymetry data. These classifications were then used to evaluate the representation and replication of seafloor structure within the MPAs and across the ecological surveys. Both the broad substrate categories and the finer scale geomorphic features were proportionately represented for many of the classes with deviations of 1-6% and 0-7%, respectively. Within MPAs, however, representation of seafloor features differed markedly from original estimates, with differences ranging up to 28%. Seafloor structure in the biological monitoring design had mismatches between sampling in the MPAs and their corresponding reference sites and some seafloor structure classes were missed entirely. The geomorphic variables derived from multibeam bathymetry data for these analyses are known determinants of the distribution and abundance of marine species and for coastal marine biodiversity. Thus, analyses like those performed in this study can be a valuable initial method of evaluating and predicting the conservation value of MPAs across a regional network.

Forests of the sea: predictive habitat modelling to assess the abundance of canopy forming kelp forests on temperate reefs

 Large brown seaweeds (kelps) form forests in temperate and boreal marine systems that serve as foundations to the structure and dynamics of communities. Mapping the distributions of these species is important to understanding the ecology of coastal environments, managing marine ecosystems (e.g., spatial planning), predicting consequences of climate change and the potential for carbon production. We demonstrate how combining seafloor mapping technologies (LiDAR and multibeam bathymetry) and models of wave energy to map the distribution and relative abundance of seaweed forests of Ecklonia radiata can provide complete coverage over hundreds of square kilometers. Using generalized linear mixed models (GLMMs), we associated observations of E. radiata abundance from video transects with environmental variables. These relationships were then used to predict the distribution of E. radiata across our 756.1km2 study area off the coast of Victoria, Australia. A reserved dataset was used to test the accuracy of these predictions. We found that the abundance distribution of E. radiata is strongly associated with depth, presence of rocky reef, curvature of the reef topography, and wave exposure. In addition, the GLMM methodology allowed us to adequately account for spatial autocorrelation in our sampling methods. The predictive distribution map created from the best GLMM predicted the abundance of E. radiata with an accuracy of 72%. The combination of LiDAR and multibeam bathymetry allowed us to model and predict E. radiata abundance distribution across its entire depth range for this study area. Using methods like those presented in this study, we can map the distribution of macroalgae species, which will give insight into ecological communities, biodiversity distribution, carbon uptake, and potential sequestration.

Environmental controls on spatial patterns in the long-term persistence of giant kelp in central California

As marine management measures increasingly protect static areas of the oceans, it is important to make sure protected areas capture and protect persistent populations. Rocky reefs in many temperate areas worldwide serve as habitat for canopy-forming macroalgae and these structure-forming species of kelps (order Laminariales) often serve as important habitat for a great diversity of species. Macrocystis pyrifera is the most common canopy-forming kelp species found along the coast of California, but the distribution and abundance of M. pyrifera varies in space and time. The purpose of this study is to determine what environmental parameters are correlated with and their relative contribution to the spatial and temporal persistence of M. pyrifera along the central coast of California and how well those environmental parameters can be used to predict areas where this species is more likely to persist. Nine environmental variables considered in this study included depth of the seafloor, structure of the rocky reef, proportion of rocky reef, size of kelp patch, biomass of kelp within a patch, distance from the edge of a kelp patch, sea surface temperature, wave orbital velocities, and population connectivity of individual kelp patches. Using a generalized linear mixed effects model (GLMM), the persistence of M. pyrifera was significantly associated with seven of the nine variables considered: depth, complexity of the rocky reef, proportion of rock, patch biomass, distance from the edge of a patch, population connectivity, and wave orbital velocities. These seven environmental variables were then used to predict the persistence of kelp across the central coast, and these predictions were compared to a reserved dataset of M. pyrifera persistence, which was not used in the creation of the GLMM. The environmental variables were shown to accurately predict the persistence of M. pyrifera within the central coast of California (r = 0.71, P < 0.001). Because persistence of giant kelp is important to the community structure of kelp forests, understanding those factors that support persistent populations of M. pyrifera will enable more effective management of these ecosystems.

Effects of human activities on rocky shores of the Surf Coast Shire: scientific basis for management at the regional level

The Surf Coast Shire in regional Victoria contains some of the most spectacular coastline in Australia, running from Point Impossible in the east to just west of the resort town of Lorne. The Surf Coast Shire council is committed to ecologically sustainable tourism based on its coastal assets, including the important intertidal environments. The challenge for the Shire is to protect and enhance the biodiversity of its intertidal areas whilst allowing for their sustainable use as a critical component of the local economy. In order to do this the Council needed to identify the conservation values of intertidal areas within the shire and assess the impacts that current human use has on these values. The impacts of shellfish collecting on rocky shores were identified as an issue of particular concern. We have conducted a research project with the Shire to provide a scientific basis for management decisions. The principal aims of this project were to: (1) determine the patterns of human use of intertidal habitats; (2) measure the impacts of human usage on biological communities and species populations; and (3) to identify intertidal sites of regional conservation significance for the Surf Coast Shire. Surveys of human usage identified reef walking, looking in rock pools and fossicking as major uses of rocky shores within the Surf Coast. This poster reports the effects of this usage on gastropod populations of rocky shores within the Surf Coast Shire. A small proportion of visitors collected intertidal organisms. Shores were categorized as high or low use based on total numbers of people observed at each shore over the first year of the project. Mean size and catch per unit effort were compared for several gastropod species between high use and low use shores. The results presented here show that the populations of some gastropod species are of smaller mean size and less abundant on high use shores than on low use shores. There was also a noticeable difference in degree of effect detected between sandstone and mudstone shores. The implications of these results are briefly discussed in terms of management options available to the Shire.

Remotely sensed hydroacoustics and observation data for predicting fish habitat suitability

This paper investigates the use of using remotely sensed observation and full coverage hydroacoustic datasets to quantify habitat suitability for a marine demersal fish, the blue-throated wrasse. Because of issues surrounding the detection of species using remotely sensed video techniques, the application of presence-only techniques are well suited for modeling demersal fish habitat suitability. Ecological-Niche Factor Analysis is used to compare analyses conducted using seafloor variables derived from hydroacoustics at three spatial scales; fine (56.25 m²), medium (506.25 m²) and coarse (2756.25 m²), to determine which spatial scale was most influential in predicting blue-throated wrasse habitat suitability. The coarse scale model was found to have the best predictive capabilities with a Boyce Index of 0.80±0.26. The global marginality and specialization values indicated that, irrespective of spatial scale, blue-throated wrasse prefer seafloor characteristics that are different to the mean available within the study site, but exhibit a relatively wide niche. Although variable importance varied over the three spatial scale models, blue-throated wrasse showed a strong preference for regions of shallow water, close to reef, with high rugosity and maximum curvature and low HSI-B values. Generally the spatial patterns in habitat suitability were well represented in the Marine National Park compared to adjacent waters. However, some significant differences in spatial patterns were observed. Interspersion and Juxtaposition Indexes for unsuitable and highly suitable habitat were significantly smaller inside the Marine National Park, while the Mean Shape Index of unsuitable habitat in the Marine National Park was significantly larger.

Field evaluation of a suite of biomarkers in an Australian Tropical Reef Species, Stripey Seaperch (Lutjanus carponotatus) : Assessment of Produced Formation water from the Harriet A Platform

There is paucity of data regarding hydrocarbon exposure of tropical fish species inhabiting the waters near oil and gas platforms on the Northwest Shelf of Australia. A comprehensive field study assessed the exposure and potential effects associated with the produced water (PW) plume from the Harriet A production platform on the northwest shelf in a local reef species, Stripey seaperch (Lutjanus carponotatus). This field study was a continuation of an earlier pilot study which concluded that there were “warning signs” of potential biological effects on fish populations exposed to PW. A 10-day field caging study was conducted deploying 15 individual fish into 6 separate steel cages set 1-m subsurface at 3 stations in a concentration gradient moving away from the platform. A battery of biomarkers were evaluated including hepatosomatic index (HSI), total cytochrome P450, bile metabolites, CYP1A-, CYP2K- and CYP2M-like proteins, cholinesterase (ChE) activity, and histopathology of liver and gill tissues. Water column and PW effluent samples was also collected. Results confirmed that PAH metabolites in bile, CYP1A-, CYP2K-, and CYP2M-like proteins and liver histopathology provided evidence of significant exposure and effects after 10 days at the near-field site (~200 m off the Harriet A platform). Hepatosomatic index, total cytochrome P450, and ChE did not provide site-specific differences by day 10 of exposure to PW. CYP proteins were shown by principal component analysis (PCA) to be the best diagnostic tool for determining exposure and associated biological effects of PW on L. carponotatus. Using a suite of biomarkers has been widely advocated as a vital component in environmental risk assessments worldwide. This study demonstrates the usefulness of biomarkers for assessing the Harriet A PW discharge into Australian waters with broader applications for other PW discharges. This approach has merit as a valuable addition to environmental management strategies for protecting Australia’s tropical environment and its rich biodiversity.

Fish assemblages associated with oil industry structures on the continental shelf of north-western Australia

This study provides the first assessment of fish associations with oil and gas structures located in deep water (85-175 m) on Australia's north-west continental shelf, using rare oil industry video footage obtained from remotely operated vehicles. A diverse range of taxa were observed associating with the structures, including reef-dependent species and transient pelagic species. Ten commercially fished species were observed, the most abundant of which was Lutjanus argentimaculatus, with an estimated biomass for the two deepest structures (Goodwyn and Echo) of 109 kg.