Experimental considerations of acute heat stress assays to quantify coral thermal tolerance

Understanding the distribution and abundance of heat tolerant corals across seascapes is imperative for predicting responses to climate change and to support novel management actions. Thermal tolerance is variable in corals and intrinsic and extrinsic drivers of tolerance are not well understood. Traditional experimental evaluations of coral heat and bleaching tolerance typically involve ramp-and-hold experiments run across days to weeks within aquarium facilities with limits to colony replication. Field-based acute heat stress assays have emerged as an alternative experimental approach to rapidly quantify heat tolerance in many samples yet the role of key methodological considerations on the stress response measured remains unresolved. Here, we quantify the effects of coral fragment size, sampling time point, and physiological measures on the acute heat stress response in adult corals. The effect of fragment size differed between species (Acropora tenuis and Pocillopora damicornis). Most physiological parameters measured here declined over time (tissue colour, chlorophyll-a and protein content) from the onset of heating, with the exception of maximum photosynthetic efficiency (Fv/Fm) which was surprisingly stable over this time scale. Based on our experiments, we identified photosynthetic efficiency, tissue colour change, and host-specific assays such as catalase activity as key physiological measures for rapid quantification of thermal tolerance. We recommend that future applications of acute heat stress assays include larger fragments (> 9 cm2) where possible and sample between 10 and 24 h after the end of heat stress. A validated high-throughput experimental approach combined with cost-effective genomic and physiological measurements underpins the development of markers and maps of heat tolerance across seascapes and ocean warming scenarios

Genes for de novo biosynthesis of omega-3 polyunsaturated fatty acids are widespread in animals

Marine ecosystems are responsible for virtually all production of omega-3 (ω3) long-chain polyunsaturated fatty acids (PUFA), which are essential nutrients for vertebrates. Current consensus is that marine microbes account for this production, given their possession of key enzymes including methyl-end (or "ωx") desaturases. ωx desaturases have also been described in a small number of invertebrate animals, but their precise distribution has not been systematically explored. This study identifies 121 ωx desaturase sequences from 80 species within the Cnidaria, Rotifera, Mollusca, Annelida, and Arthropoda. Horizontal gene transfer has contributed to this hitherto unknown widespread distribution. Functional characterization of animal ωx desaturases provides evidence that multiple invertebrates have the ability to produce ω3 PUFA de novo and further biosynthesize ω3 long-chain PUFA. This finding represents a fundamental revision in our understanding of ω3 long-chain PUFA production in global food webs, by revealing that numerous widespread and abundant invertebrates have the endogenous capacity to make significant contributions beyond that coming from marine microbes. Copyright © 2018 The Authors, some rights reserved.Acknowledgments: We thank A. Magurran and J. Napier for comments on the manuscript and R. Ruivo for drawings in Figs. 1 and 3. Funding: This work received funding from the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland) funded by the Scottish Funding Council (grant reference HR09011), and their support is gratefully acknowledged. Access to the Institute of Aquaculture laboratories was funded by the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 262336 (AQUAEXCEL), Transnational Access Project Number 0095/06/03/13

Measurement of the Atmospheric Muon Spectrum from 20 to 3000 GeV

The absolute muon flux between 20 GeV and 3000 GeV is measured with the L3 magnetic muon spectrometer for zenith angles ranging from 0 degree to 58 degree. Due to the large exposure of about 150 m2 sr d, and the excellent momentum resolution of the L3 muon chambers, a precision of 2.3 % at 150 GeV in the vertical direction is achieved. The ratio of positive to negative muons is studied between 20 GeV and 500 GeV, and the average vertical muon charge ratio is found to be 1.285 +- 0.003 (stat.) +- 0.019 (syst.).Comment: Total 32 pages, 9Figure

Habitat selection and aggression as determinants of spatial segregation among damselfish on a coral reef

Adults of many closely related coral reef fish species are segregated along gradients of depth or habitat structure. Both habitat selection by new settlers and subsequent competitive interactions can potentially produce such patterns, but their relative importance is unclear. This study examines the potential roles of habitat selection and aggression in determining the spatial distribution of adults and juveniles of four highly aggressive damselfishes at Lizard Island, northern Great Barrier Reef. Dischistodus perspicillatus, D. prosopotaenia, D. melanotus, and D. pseudochrysopoecilus maintain almost non-overlapping distributions across reef zones, with adults of one species dominating each reef zone. Juveniles exhibit slightly broader distributional patterns suggesting that subsequent interactions reduce overlap among species. Although habitat choice experiments in aquaria suggest that associations between juveniles and substrata types in the field are partly due to habitat selection, large overlaps in the use of substrata by the different species were also found, suggesting that substratum selection alone is insufficient in explaining the discrete spatial distributions of adults. The strength of aggressive interactions among all four species was tested by a "bottle" experiment, in which an adult or juvenile of each species was placed in the territories of adult fish on the reef. The greatest levels of interspecific aggression were directed against adults and juveniles of neighbouring species. The highest levels of aggression were associated with species exhibiting the greatest levels of overlap in resource use. Evidently both habitat selection and interspecific aggression combine to determine the adult distributions of these species

Intraspecific variation in the pelagic larval duration of tropical reef fishes

Estimates of pelagic larval duration (PLD) for 10 species of Pomacentridae and two species of Gobiidae were made. In eight of the 12 species examined, within-population mean PLDs differed between sampling times, locations within regions and among regions. In contrast, the range of these same PLD estimates overlapped at all spatial and temporal scales examined in 11 of the 12 species, but not between regions in one species (Amphiprion melanopus). Therefore, despite tight error estimates typically associated with estimates of PLD taken from a particular population at a particular time in some taxa, the overlapping ranges in PLD reported here indicate that the length of the pelagic larval phase is a much more plastic trait than previously appreciated. Within-species variation in PLD has considerable potential to provide further insights into the ecology and evolution of tropical reef fishes

Intraspecific variation in the pelagic larval duration of tropical reef fishes

Estimates of pelagic larval duration (PLD) for 10 species of Pomacentridae and two species of Gobiidae were made. In eight of the 12 species examined, within-population mean PLDs differed between sampling times, locations within regions and among regions. In contrast, the range of these same PLD estimates overlapped at all spatial and temporal scales examined in 11 of the 12 species, but not between regions in one species (Amphiprion melanopus). Therefore, despite tight error estimates typically associated with estimates of PLD taken from a particular population at a particular time in some taxa, the overlapping ranges in PLD reported here indicate that the length of the pelagic larval phase is a much more plastic trait than previously appreciated. Within-species variation in PLD has considerable potential to provide further insights into the ecology and evolution of tropical reef fishes

Microsatellite allele sizes alone are insufficient to delineate species boundaries in Symbiodinium

Symbiodinium are a diverse group of unicellular dinoflagellates that are important nutritional symbionts of reef-building corals. Symbiodinium putative species ('types') are commonly identified with genetic markers, mostly nuclear and chloroplast encoded ribosomal DNA regions. Population genetic analyses using microsatellite loci have provided insights into Symbiodinium biogeography, connectivity and phenotypic plasticity, but are complicated by: (i) a lack of consensus criteria used to delineate inter- vs. intragenomic variation within species; and (ii) the high density of Symbiodinium in host tissues, which results in single samples comprising thousands of individuals. To address this problem, Wham & LaJeunesse (2016) present a method for identifying cryptic Symbiodinium species from microsatellite data based on correlations between allele size distributions and nongeographic genetic structure. Multilocus genotypes that potentially do not recombine in sympatry are interpreted as secondary 'species' to be discarded from downstream population genetic analyses. However, Symbiodinium species delineations should ideally incorporate multiple physiological, ecological and molecular criteria. This is because recombination tests may be a poor indicator of species boundaries in Symbiodinium due to their predominantly asexual mode of reproduction. Furthermore, discontinuous microsatellite allele sizes in sympatry may be explained by secondary contact between previously isolated populations and by mutations that occur in a nonstepwise manner. Limitations of using microsatellites alone to delineate species are highlighted in earlier studies that demonstrate occasional bimodal distributions of allele sizes within Symbiodinium species and considerable allele size sharing among Symbiodinium species. We outline these issues and discuss the validity of reinterpretations of our previously published microsatellite data from Symbiodinium populations on the Great Barrier Reef (Howells et al. 2013)

High genetic diversities and complex genetic structure in an Indo-Pacific tropical reef fish (Chlorurus sordidus): evidence of an unstable evolutionary past?

Historical sea level fluctuations have influenced the genetic structure and evolutionary history of marine species and examining widespread species across their speciesrsquo ranges may elucidate some of these effects. Chlorurus sordidus is a common and widespread parrotfish found on coral reefs throughout the Indo-central Pacific. We used phylogenetic, phylogeographic, and cladistic analyses to examine the genetic composition and population structure of this species across most of its latitudinal range limits. We sequenced 354 bp of the mitochondrial control region I in 185 individuals from nine populations. Populations of C. sordidus displayed high levels of genetic diversity, similar to those recorded for widespread pelagic fish species, but much greater nucleotide diversity values than those previously recorded for other demersal reef fishes. Both phylogenetic and phylogeographic analyses detected strong genetic subdivision at the largest spatial scale (i.e. among oceans). The Pacific Ocean was characterised by weak population genetic structure. Separation of the Hawaiian location from other Pacific and West Indian Ocean sites was evident in phylogenetic analyses, but not from analysis of molecular variance. NCA and isolation-by-distance tests suggested that the genetic structure of this species was the result of multiple contemporary and historical processes, including long-distance colonisation and range expansion arising from fluctuating sea levels, limited current gene flow, and isolation by distance. This pattern is to be expected when historically fragmented populations come into secondary contact. We suggest the patterns of population genetic structure recorded in C. sordidus are caused by large local population sizes, high gene flow, and a recent history of repeated fragmentation and remixing of populations resulting from fluctuating sea levels

Biennium horribile: very high mortality in the reef coral Acropora millepora on the Great Barrier Reef in 2009 and 2010

Coral cover has declined markedly in the recent past in many regions of the world, including the Great Barrier Reef (GBR), Australia. The major causes of this decline are generally considered to be mortality associated with large-scale severe disturbances (i.e. catastrophic mortality), such as Acanthaster planci outbreaks, cyclones and bleaching. However, background rates of mortality (i.e. not associated with catastrophic disturbance), are rarely quantified, but without these it is difficult to assess the relative importance of these 2 types of mortality (catastrophic and background). We quantified spatial and temporal variation in catastrophic and background whole-colony mortality of the common reef coral Acropora millepora over 24 mo at 2 sites in 3 regions separated by 700 km along the GBR. The study period included 2 cyclones and a flood. Overall mortality rates were exceptionally high. Of 180 colonies tagged in April 2009, only 36 (20%) were alive in April 2011, and 68% of this mortality occurred in intervals following the 3 large disturbances. Background mortality rates were also high in the Palm Islands, where they approached 40% yr-1 compared to <5% in the Whitsunday and Keppel Islands. These results support the hypothesis that catastrophic mortality has been the major cause of coral loss in recent years on the GBR and also suggest that background rates of mortality are increasing at some locations. Projected increases in the agents of catastrophic mortality, such as cyclones and bleaching, as a result of global warming are likely to threaten the persistence of many coral species