Fluorescence in situ hybridization and spectral imaging of coral-associated bacterial communities

Microbial communities play important roles in the functioning of coral reef communities. However, extensive autofluorescence of coral tissues and endosymbionts limits the application of standard fluorescence in situ hybridization (FISH) techniques for the identification of the coral-associated bacterial communities. This study overcomes these limitations by combining FISH and spectral imaging.

Genetic structure of a reef-building coral from thermally distinct environments on the Great Barrier Reef

Adaptation to localised thermal regimes is facilitated by restricted gene flow, ultimately leading to genetic divergence among populations and differences in their physiological tolerances. Allozyme analysis of six polymorphic loci was used to assess genetic differentiation between nine populations of the reef-building coral Acropora millepora over a latitudinal temperature gradient on the inshore regions of the Great Barrier Reef (GBR). Small but significant genetic differentiation indicative of moderate levels of gene flow (pairwise F-ST 0.023 to 0.077) was found between southern populations of A. millepora in cooler regions of the GBR and the warmer, central or northern GBR populations. Patterns of genetic differentiation at these putatively neutral allozyme loci broadly matched experimental variation in thermal tolerance and were consistent with local thermal regimes (warmest monthly-averages) for the A. millepora populations examined. It is therefore hypothesized that natural selection has influenced the thermal tolerance of the A. millepora populations examined and greater genetic divergence is likely to be revealed by examination of genetic markers under the direct effects of natural selection.

Increased mortality and photoinhibition in the symbiotic dinoflagellates of the Indo-Pacific coral Stylophora pistillata (Esper) after summer bleaching

Coral bleaching (the loss of symbiotic dinoflagellates from reef-building corals) is most frequently caused by high-light and temperature conditions. We exposed the explants of the hermatypic coral Stylophora pistillata to four combinations of light and temperature in late spring and also in late summer. During mid-summer, two NOAA bleaching warnings were issued for Heron Island reef (Southern Great Barrier Reef, Australia) when sea temperature exceeded the NOAA bleaching threshold, and a 'mild' (in terms of the whole coral community) bleaching event occurred, resulting in widespread S. pistillata bleaching and mortality. Symbiotic dinoflagellate biomass decreased by more than half from late spring to late summer (from 2.5x10(6) to 0.8x10(6) dinoflagellates cm(2) coral tissue), and those dinoflagellates that remained after summer became photoinhibited more readily (dark-adapted F (V) : F (M) decreased to (0.3 compared with 0.4 in spring), and died in greater numbers (up to 17% dinoflagellate mortality compared with 5% in the spring) when exposed to artificially elevated light and temperature. Adding exogenous antioxidants (D-mannitol and L-ascorbic acid) to the water surrounding the coral had no clear effect on either photoinhibition or symbiont mortality. These data show that light and temperature stress cause mortality of the dinoflagellate symbionts within the coral, and that susceptibility to light and temperature stress is strongly related to coral condition. Photoinhibitory mechanisms are clearly involved, and will increase through a positive feedback mechanism: symbiont loss promotes further symbiont loss as the light microenvironment becomes progressively harsher.

Mass mortality following disturbance in Holocene coral reefs from Papua New Guinea

The frequency and intensity of disturbance on living coral reefs have been accelerating for the past few decades, resulting in a changed seascape. What is unclear but vital for management is whether this acceleration is natural or coincident only with recent human impact. We surveyed nine uplifted early to mid-Holocene (11,000-3700 calendar [cal] yr B.P.) fringing and barrier reefs along similar to 27 km at the Huon Peninsula, Papua New Guinea. We found evidence for several episodes of coral mass mortality, but frequency was < 1 in 1500 yr. The most striking mortality event extends > 16 km along the ancient coastline, occurred ca. 9100-9400 cal yr B.P., and is associated with a volcanic ash horizon. Recolonization of the reef surface and resumption of vertical reef accretion was rapid (< 100 yr), but the post-disturbance reef communities contrasted with their pre-disturbance counterparts. Assessing the frequency, nature, and long-term ecological consequences of mass-mortality events in fossil coral reefs may provide important insights to guide management of modern reefs in this time of environmental degradation and change.

Monitoring coral bleaching using a colour reference card

Assessment of the extent of coral bleaching has become an important part of studies that aim to understand the condition of coral reefs. In this study a reference card that uses differences in coral colour was developed as an inexpensive, rapid and non-invasive method for the assessment of bleaching. The card uses a 6 point brightness/saturation scale within four colour hues to record changes in bleaching state. Changes on the scale of 2 units or more reflect a change in symbiont density and chlorophyll a content, and therefore the bleaching state of the coral. When used by non-specialist observers in the field (here on an intertidal reef flat), there was an inter-observer error of I colour score. This technique improves on existing subjective assessment of bleaching state by visual observation and offers the potential for rapid, wide-area assessment of changing coral condition.

Response of holosymbiont pigments from the scleractinian coral Montipora monasteriata to short-term heat stress

Heating the scleractinian coral, Montipora monasteriata (Forskal 1775) to 32 degrees C under < 650 mu mol quanta m(-2) s(-1) led to bleaching in the form of a reduction in Peridinin, xanthophyll pool, chlorophyll c(2) and chlorophyll a, but areal dinoflagellates densities did not decline. Associated with this bleaching, chlorophyll (Chl) allomerization and dinoflagellate xanthophyll cycling increased. Chl allomerization is believed to result from the interaction of Chl with singlet oxygen (O-1(2)) or other reactive oxygen species. Thermally induced increases in Chl allomerization are consistent with other studies that have demonstrated that thermal stress generates reactive oxygen species in symbiotic dinoflagellates. Xanthophyll cycling requires the establishment of a pH gradient across the thylakoid membrane. Our results indicate that, during the early stages of thermal stress, thylakoid membranes are intact. Different morphs of M. monasteriata responded differently to the heat stress applied: heavily pigmented coral hosts taken from a high-light environment showed significant reductions in green fluorescent protein (GFP)-like homologues, whereas nonhost pigmented high-light morphs experienced a significant reduction in water-soluble protein content. Paradoxically, the more shade acclimated cave morph were, based on Chl fluorescence data, less thermally stressed than either of the high-light morphs. These results Support the importance of coral pigments for the regulation of the light environment within the host tissue.

Spatial patterns of aerobic and anaerobic mineralization rates and oxygen penetration dynamics in coral reef sediments

Oxygen consumption rates (OCR), aerobic mineralization and sulfate reduction rates (SRR) were studied in the permeable carbonate reef sediments of Heron Reef, Australia. We selected 4 stations with different hydrodynamic regimes for this study. In situ oxygen penetration into the sediments was measured with an autonomous microsensor profiler. Areal OCR were quantified from the measured oxygen penetration depth and volumetric OCR. Oxygen penetration and dynamics (median penetration depths at the 4 stations ranged between 0.3 and 2.2 cm), OCR (median 57 to 196 mmol C m(-2) d(-1)), aerobic mineralization (median 24 to 176 mmol C m(-2) d(-1)) and SRR (median 9 to 42 mmol C m(-2) d(-1)) were highly variable between sites. The supply of oxygen by pore water advection was a major cause for high mineralization rates by stimulating aerobic mineralization at all sites. However, estimated bottom water filtration rates could not explain the differences in volumetric OCR and SRR between the 4 stations. This suggests that local mineralization rates are additionally controlled by factors other than current driven pore water advection, e.g. by the distribution of the benthic fauna or by local differences in labile organic carbon supply from sources such as benthic photosynthesis. Carbon mineralization rates were among the highest reported for coral reef sediments, stressing the role of these sediments in the functioning of the reef ecosystem.

The impact of spectral composition and light periodicity on the activity of two antioxidant enzymes (SOD and CAT) in the coral Favia favus

In oligotrophic waters the light spectrum is mostly blue, and therefore the physiological and biochemical responses to blue light occurring in the coral tissue and in the symbiotic algae are important. Examination of the wavelength dependence of two free radical scavenger enzyme activity revealed an increase in activity in the blue light range (440-480 nm) compared to the red (640680 nm) in the full visible light (400-700 nm) range. These data show for the first time the relationship between the action spectra of photosynthesis and the activity of two main antioxidant enzymes in the symbiotic coral Favia favus. It was found that in the animal (host) the enzyme response to the spectral distribution of light was higher than that of the zooxanthellae, probably due to accumulation of free radicals within the host tissue. Furthermore, we found that the activity of these enzymes is affected in nature by the length of the day and night, and in the laboratory, by the duration of the illumination. Changes in the pigment concentrations were also observed in response to growth under the blue region and the whole PAR spectrum, while fluorescence measurements with the fast repetition rate fluorometer (FRRF) showed a decrease in the sigma cross section and a decrease in the quantum yield also in the blue part of the spectrum. These changes of scavenger enzymes activity, pigment concentration and fluorescence yield at different light spectra are vital in acclimatization and survival of corals in shallow water environments with high light radiation. (c) 2005 Elsevier B.V. All rights reserved.

Why are there so few genetic markers available for coral population analyses?

Coral reefs are in serious decline, and research in support of reef management objectives is urgently needed. Reef connectivity analyses have been highlighted as one of the major future research avenues necessary for implementing effective management initiatives for coral reefs. Despite the number of new molecular genetic tools and the wealth of information that is now available for population-level processes in many marine disciplines, scleractinian coral population genetic information remains surprisingly limited. Here we examine the technical problems and approaches used, address the reasons contributing to this delay in understanding, and discuss the future of coral population marker development. Considerable resources are needed to target the immediate development of an array of relevant genetic markers coupled with the rapid production of management focused data in order to help conserve our globally threatened coral reef resources.

Phototrophic microendoliths bloom during coral "white syndrome"

Following rapid lesion progression of white syndrome in tabular Acropora spp., the white bare skeleton gradually changes to green, a result of endolithic algae blooms (primarily Ostreobium spp.). Endolithic algal biomass and chlorophyll concentration were found to be an order of magnitude higher in the green zone compared with healthy appearing parts of each colony. Chl b to Chl a ratio increased from 1:1.6 in the healthy area to 1:2 and 1:3.5 in the white exposed skeleton and green zones, respectively. These observations together with pulse amplitude modulated (PAM) fluorometry suggest photoacclimation of the endoliths in the green zone. Histopathological microscopy revealed that the endolithic algal filaments penetrate the coral tissue. This study highlights the interaction of endolithic algae with both the skeleton and host tissue. This may have a critical role in the processes that accompany the post-disease state in reef-building corals.

A high-precision record of mid-late Holocene sea-level events from emergent coral pavements in the Houtman Abrolhos Islands, southwest Australia

Early work on sea-levels in southwest Australia claimed to recognise a Holocene sea-level highstand which was not seen in better known sea-level records elsewhere at the time. More recent work has confirmed that a mid-Holocene highstand Occurred about 6 kyr ago. As new data on oscillating sea-levels from the region have recently been published, a high continuity, precisely dated and accurately surveyed record was obtained from emergent coral pavements in the leeward Houtman Abrolhos Islands (Serventy Island), a tectonically stable region from where good-quality Holocene sea-level data have been previously obtained from corals. From the mid-Holocene highstand ca. 7 U/Th kyr ago, sea-level declined linearly during the remainder of the Holocene as the carbonate platform prograded leewards. Hydro-isostatic controls are probably significant in the record. (c) 2005 Elsevier Ltd and INQUA. All rights reserved.

High-precision TIMS U-series and AMS C-14 dating of a coral reef lagoon sediment core from southern South China Sea

Accurate dating of lagoon sediments has been a difficult problem, although lagoon profiles, usually with high deposition rates, have a great potential for high-resolution climate reconstruction. We report 26 high-precision TIMS U-series dates (on 25 coral branches) and five AMS C-14 dates (on foraminifera) for a 15.4-m long lagoon core from Yongshu Reef, Nansha area, southern South China Sea. All the dates are in the correct stratigraphical sequence, providing the best chronology so far reported for lagoon deposits. The results reveal a similar to 4000-a continuous depositional history, with sedimentation rates varying from 0.8 to 24.6 mm a(-1), with an average of 3.85 mm a(-1), which corresponds to an average net carbonate accumulation rate of similar to 2700 g CaCO3 m(-2) a(-1), significantly higher than the mean value (800 +/- 400 g CaCO3 m(-2) a(-1)) used for lagoons in general in previous studies of global carbonate budget. Episodes of accelerated depositions within the last 1000 years correlate well with strong storm events identified by U-series dates of storm-transported coral blocks in the area. However, in the longer term, the sedimentation rates during the past 1000 years were much higher than earlier on, probably due to more vigorous wave-reef interaction as a result of relative sea-level fall since 500 AD and expansion of reef flat area, supplying more sediments. The coral TIMS U-series ages and foraminifera AMS 14C dates reveal intriguing apparent radiocarbon reservoir ages (R) from 572 to 1052 years, which are much higher than global mean values of similar to 400 years. (c) 2006 Elsevier Ltd. All rights reserved.

The stability of P in coral reef fishes

The constancy of phenotypic variation and covariation is an assumption that underlies most recent investigations of past selective regimes and attempts to predict future responses to selection. Few studies have tested this assumption of constancy despite good reasons to expect that the pattern of phenotypic variation and covariation may vary in space and time. We compared phenotypic variance-covariance matrices (P) estimated for Populations of six species of distantly related coral reef fishes sampled at two locations on Australia's Great Barrier Reef separated by more than 1000 km. The intraspecific similarity between these matrices was estimated using two methods: matrix correlation and common principal component analysis. Although there was no evidence of equality between pairs of P, both statistical approaches indicated a high degree of similarity in morphology between the two populations for each species. In general, the hierarchical decomposition of the variance-covariance structure of these populations indicated that all principal components of phenotypic variance-covariance were shared but that they differed in the degree of variation associated with each of these components. The consistency of this pattern is remarkable given the diversity of morphologies and life histories encompassed by these species. Although some phenotypic instability was indicated, these results were consistent with a generally conserved pattern of multivariate selection between populations.

Viruses: agents of coral disease?

The potential role of viruses in coral disease has only recently begun to receive attention. Here we describe our attempts to determine whether viruses are present in thermally stressed corals Pavona danai, Acropora formosa and Stylophora pistillata and zoanthids Zoanthus sp., and their zooxanthellae. Heat-shocked P. danai, A. formosa and Zoanthus sp. all produced numerous virus-like particles (VLPs) that were evident in the animal tissue, zooxanthellae and the surrounding seawater; VLPs were also seen around heat-shocked freshly isolated zooxanthellae (FIZ) from P. danai and S. pistillata. The most commonly seen VLPs were tail-less, hexagonal and about 40 to 50 nm in diameter, though a diverse range of other VLP morphotypes (e.g. rounded, rod-shaped, droplet-shaped, filamentous) were also present around corals. When VLPs around heat-shocked FIZ from S. pistillata were added to non-stressed FIZ from this coral, they resulted in cell lysis, suggesting that an infectious agent was present; however, analysis with transmission electron microscopy provided no clear evidence of viral infection. The release of diverse VLPs was again apparent when flow cytometry was used to enumerate release by heat-stressed A. formosa nubbins. Our data support the infection of reef corals by viruses, though we cannot yet determine the precise origin (i.e. coral, zooxanthellae and/or surface microbes) of the VLPs seen. Furthermore, genome sequence data are required to establish the presence of viruses unequivocally.