Impacts of bleaching on the soft coral Lobophytum compactum. I. Fecundity, fertilization and offspring viability

We document long-term effects of a simulated bleaching event on the reproductive output and offspring viability of the soft coral Lobophytum compactum. Corals were subjected to temperature and solar radiation treatments to produce both moderately (48–60%) and heavily (90–95%) bleached colonies. Although bleached colonies recovered their zooxanthellae within 10 to 18 weeks, impacts on reproductive output were significant for at least two annual spawning seasons. In the first year, both polyp fecundity and mean oocyte diameter were reduced and inversely correlated with the degree of bleaching, with complete failure of fertilization in the group of heavily bleached colonies. For moderately bleached soft corals, survival and growth of sexual offspring did not differ significantly from those of unbleached colonies. Although no further reductions in zooxanthellae densities in experimental soft corals were recorded throughout the subsequent second year, egg size and fecundity of the heavily bleached soft corals were still significantly reduced 20 months later. Severe bleaching clearly has long-term sub-lethal impacts, reducing overall reproductive output for at least two spawning seasons. Accepted: 1 June 2000     

Scleractinian coral diseases in south Florida: incidence, species susceptibility, and mortality

There are limited quantitative data available documenting the natural, or non-epizootic, occurrence of scleractinian coral diseases over multiple years. Individual coral colonies exhibiting black band disease (BBD), white plague (WP), dark spots syndrome (DSS), and white band disease (WBD) were monitored 3 times per year on 5 south Florida reefs over a 2 yr period. Surveys included measurements of coral population composition, coral diversity, disease type, coral species affected, colony size, percent of colony affected, and the number of lesions or active infections per colony. Data on re-infections of the same colonies, multiple infections per colony, disease duration, disease-associated tissue mortality, and coral recruitment are also presented. A total of 674 coral colonies exhibiting coral diseases were tagged and monitored. DSS was the most common syndrome (n = 620 infected colonies), but BBD and WP infections caused the largest amount of coral tissue death. The only disease that exhibited a linear increase in incidence with elevated temperature was BBD. DSS and BBD were the most persistent conditions, and WP infections were comparatively short-lived, with obvious signs of disease typically disappearing after 2 to 3 mo. The only disease that caused total colony death as oppposed to partial mortality during the survey period was WBD. WP and DSS incidence was significantly positively correlated with the relative frequency of the species most commonly affected by each disease at each study site. Of the 61 colonies examined in the recruitment survey, only 5 scleractinian coral recruits were identified. The most commonly recorded colonizer of exposed coral skeleton was filamentous/turf algae, thus indicating the potential for a shift towards algal-dominated reef communities.     

Experimental fragmentation reduces sexual reproductive output by the reef-building coral Pocillopora damicornis

Natural and anthropogenic disturbances may fragment stony reef corals, but few quantitative data exist on the impacts of skeletal fragmentation on sexual reproduction in corals. We experimentally fragmented colonies of the branching coral Pocillopora damicornis and determined the number and size of planula larvae released during one lunar reproductive cycle. Partially fragmented colonies significantly delayed both the onset and peak period of planula release compared with intact control colonies. Most fragments removed from the corals died within 11–18 days, and released few planulae. The total number of planulae released per coral colony varied exponentially with remaining tissue volume, and was significantly lower in damaged versus undamaged colonies. However, the number of planulae produced per unit tissue volume, and planula size, did not vary with damage treatment. We conclude that even partial fragmentation of P. damicornis colonies (<25% of tissue removed) decreases their larval output by reducing reproductive tissue volume. Repeated breakage of corals, such as caused by intensive diving tourism or frequent storms, may lead to substantially reduced sexual reproduction. Therefore, reef management should limit human activities that fracture stony corals and lead to decreases in colony size and reproductive output. Accepted: 2 February 2000     

Three scleractinian coral diseases in Dominica, West Indies: distribution, infection patterns and contribution to coral tissue mortality

Coral diseases have been documented in many areas of the Caribbean, but studies in the eastern Caribbean region have been lacking. The prevalence, distribution patterns and contribution to the mortality of coral tissue by black band discase (BBD), white plague (WP) and dark spots disease (DSD) were examined at five reef sites along the west coast of Dominica. 185 of the 325 diseased colonies recorded between March and August 2000, in a survey area of 5884 m2, were WP. This disease contributed to 89% of the total 4.08 m2 of tissue mortality caused by diseases during the survey period. WP also affected the largest average tissue surface area (relative to colony size) per colony and exhibited the largest average tissue loss per infection when compared to BBD and DSD. The species most susceptible to WP and BBD in Dominica differed from most other described Caribbean locations with Siderastrea siderea being most susceptible. S. siderea was also the only species noted to be susceptible to DSD. Measurements of colony size revealed that each disease affected the larger colonies of some coral species. Comparisons between disease prevalence at each site and various physical parameters, including temperature, wave height, depth, and current patterns, did not exhibit significant correlations. The lack of a direct correlation between temperature and disease prevalence indicates that there are other seasonal factors contributing to the higher prevalence of diseases recorded during the summer months in Dominica. WP prevalence at each site was positively correlated to the relative species abundances of the species most susceptible to WP. This was the dominant factor in determining site-specific disease densities of this disease and may therefore be a valuable predictive and management tool. There were no correlations between BBD or DSD and the relative abundances of susceptible species. The spatial distribution patterns of WP, BBD and DSD were clustered, which is a distribution pattern that suggests an infectious disease.     

Intraspecific contact and egg production in the massive coral Goniastrea aspera in Okinawa, subtropical Japan

The effect of intraspecific contact (Contact) on egg production was examined in the massive coral Goniastrea aspera in Okinawa, subtropical Japan. The contact was non-aggressive without damaging soft tissues each other. Within Contact colonies, polyp volume, polyp fertility (%polyps with gonad), and NE/PV (number of eggs per polyp volume) were significantly smaller in marginal (Mg) polyps without direct intraspecific contact than other polyps, but no difference was found between non-marginal and Mg-Contact (marginal with direct intraspecific contact) polyps. Comparisons of non-marginal polyps (non-marginal and Mg-Contact polyps were combined in Contact colonies) between Non-Contact and Contact colonies showed that fertility and NE/PV were significantly larger in Contact colonies than in Non-Contact colonies, but polyp volume were not different significantly. Further analyses dividing colonies at Non-Contact maturation colony size (60 polyps) revealed that fertility and NE/PV were significantly larger in Contact colonies than in Non-Contact colonies only in the small colonies (<60 polyps), indicating that the intraspecific contact promoted sexual maturation at smaller colony size; one polyped Contact coral was also reproductive. The lack of correlation between polyp volume and NE/PV in the small Contact colonies, and the similarity of NE/PV in non-marginal and Mg-Contact polyps within a colony, suggest that the maturation at smaller size in Contact colonies is realized by reproductive integration of polyps at the colony level. The present results show that size-structured populations such as colonial corals may show phenotypic diversity in key demographic parameters, such as reproductive output, dependent on ecological conditions.     

The bacterial ecology of a plague-like disease affecting the Caribbean coral Montastrea annularis

The bacterial communities associated with the Caribbean coral Montastrea annularis showing tissue lesions indicative of a White Plague (WP)-like disease were investigated. Two molecular screening techniques using bacterial 16S rDNA genes were used and demonstrated distinct differences between the bacterial communities of diseased and non-diseased coral tissues, and also in relation to the proximity of tissue lesions on diseased corals. Differences between non-diseased corals and the apparently healthy tissues remote from the tissue lesion in affected corals indicates a 'whole coral' response to a relatively small area of infection with a perturbation in the normal microbial flora occurring prior to the onset of visible signs of disease. These whole organism changes in the microbial flora may serve as a bioindicator of environmental stress and disease. There were striking similarities between the 16S rDNA sequence composition associated with the WP-like disease studied here and that previously reported in association with black band disease (BBD) in coral. Similarities included the presence of a potential pathogen, an alpha-proteobacterium identified as the causal agent of juvenile oyster disease (JOD). The WP-like disease studied here is apparently different to WP Type ii because the bacterial species previously identified as the causal agent of WP Type ii was not detected, although the symptoms of the two diseases are similar.     

Sexual Reproduction and Seasonality of the Alaskan Red Tree Coral,Primnoa pacifica

The red tree coral Primnoa pacifica is an important habitat forming octocoral in North Pacific waters. Given the prominence of this species in shelf and upper slope areas of the Gulf of Alaska where fishing disturbance can be high, it may be able to sustain healthy populations through adaptive reproductive processes. This study was designed to test this hypothesis, examining reproductive mode, seasonality and fecundity in both undamaged and simulated damaged colonies over the course of 16 months using a deepwater-emerged population in Tracy Arm Fjord. Females within the population developed asynchronously, though males showed trends of synchronicity, with production of immature spermatocysts heightened in December/January and maturation of gametes in the fall months. Periodicity of individuals varied from a single year reproductive event to some individuals taking more than the 16 months sampled to produce viable gametes. Multiple stages of gametes occurred in polyps of the same colony during most sampling periods. Mean oocyte size ranged from 50 to 200 µm in any season, and maximum oocyte size (802 µm) suggests a lecithotrophic larva. No brooding larvae were found during this study, though unfertilized oocytes were found adhered to the outside of polyps, where they are presumably fertilized. This species demonstrated size-dependent reproduction, with gametes first forming in colonies over 42-cm length, and steady oocyte sizes being achieved after reaching 80-cm in length. The average fecundity was 86 (±12) total oocytes per polyp, and 17 (±12) potential per polyp fecundity. Sub-lethal injury by removing 21–40% of colony tissue had no significant reproductive response in males or females over the course of this study, except for a corresponding loss in overall colony fecundity. The reproductive patterns and long gamete generation times observed in this study indicate that recruitment events are likely to be highly sporadic in this species increasing its vulnerability to anthropogenic disturbances.     

Coral-crab association: a compact domain of a multilevel trophic system

Colonies of the Red Sea branching coral Stylophora pistillata were incubated in situ with radioactive carbon and Trapezia cymodoce crabs were introduced to the colonies, for one month each, up to 7 months after coral labelling. Zooxanthellar photosynthetic products were translocated to the crabs via host coral tissue. Based upon crab/coral tissue conversion from 53 crab samples, crabs possessed radioactive material equal to that recorded in 320–770 mm² of coral tissue (up to 2257 mm²). This material was translocated mainly by direct grazing on coral tissue rather than mucus collection. Ovigerous female crabs (39 specimens) accumulated significantly more labelled carbon than males, and up to 53 % of their radioactivity was concentrated in their reproductive organs. A pair of crabs dwelling in a coral colony consumed ca 130 cm² of host tissue per month (40–45 cm length of coral branches). This system represents a compact, obligatory multilevel trophic domain which also radiates horizontally by allocation of energy derived from algal photosynthesis into planula and zooea larva production, permitting the development of long trophic chains and a complex food web.     

Potential role of viruses in white plague coral disease

White plague (WP)-like diseases of tropical corals are implicated in reef decline worldwide, although their etiological cause is generally unknown. Studies thus far have focused on bacterial or eukaryotic pathogens as the source of these diseases; no studies have examined the role of viruses. Using a combination of transmission electron microscopy (TEM) and 454 pyrosequencing, we compared 24 viral metagenomes generated from Montastraea annularis corals showing signs of WP-like disease and/or bleaching, control conspecific corals, and adjacent seawater. TEM was used for visual inspection of diseased coral tissue. No bacteria were visually identified within diseased coral tissues, but viral particles and sequence similarities to eukaryotic circular Rep-encoding single-stranded DNA viruses and their associated satellites (SCSDVs) were abundant in WP diseased tissues. In contrast, sequence similarities to SCSDVs were not found in any healthy coral tissues, suggesting SCSDVs might have a role in WP disease. Furthermore, Herpesviridae gene signatures dominated healthy tissues, corroborating reports that herpes-like viruses infect all corals. Nucleocytoplasmic large DNA virus (NCLDV) sequences, similar to those recently identified in cultures of Symbiodinium (the algal symbionts of corals), were most common in bleached corals. This finding further implicates that these NCLDV viruses may have a role in bleaching, as suggested in previous studies. This study determined that a specific group of viruses is associated with diseased Caribbean corals and highlights the potential for viral disease in regional coral reef decline.     

Coral diseases near Lee Stocking Island, Bahamas: patterns and potential drivers

The number of coral diseases, coral species they infect, number of reported cases, and range over which these diseases are distributed have all increased dramatically in the past 3 decades, posing a serious threat to coral reef ecosystems worldwide. While some published studies provide data on the distribution of coral diseases at local and regional levels, few studies have addressed the factors that may drive these distributions. We recorded coral disease occurrence, prevalence, and severity along with temperature, sedimentation, and coral population data (species abundance and colony size) over 2 consecutive summers on reefs near Lee Stocking Island (LSI) in the Bahamas' Exuma Chain. In 2002 a total of 11092 coral colonies (all species present) were examined within a survey area of 9420 m2, and 13 973 colonies within 10 362 m2 in 2003. Similar to other reports, relatively large, framework species including Siderastrea siderea, Colpophyllia natans, and Montastraea annularis, along with the smaller Dichocoenia stokesi, were the species most susceptible to coral disease. Recurring infections were observed on individual colonies from 2002 to 2003, and were more likely for black band disease (BBD) than for either white plague (WP) or dark spots syndrome (DS). In 2002, WP and DS demonstrated clumped distributions, while BBD was randomly distributed. However, in 2003 BBD and WP were clumped. This is the first study, to our knowledge, that quantitatively documents coral disease dynamics on reefs surrounding LSI.     

Gametogenesis and fecundity of <Emphasis Type="Italic">Acropora tenella</Emphasis> (Brook 1892) in a mesophotic coral ecosystem in Okinawa,Japan

Mesophotic coral ecosystems (below 30–40 m depth) host a large diversity of zooxanthellate coral communities and may play an important role in the ecology and conservation of coral reefs. Investigating the reproductive biology of mesophotic corals is important to understand their life history traits. Despite an increase in research on mesophotic corals in the last decade, their reproductive biology is still poorly understood. Here, gametogenesis and fecundity of the Indo-Pacific mesophotic coral, Acropora tenella, were examined in an upper mesophotic reef (40 m depth) in Okinawa, Japan for the first time. Acropora tenella is a hermaphrodite with a single annual gametogenic cycle, and both oogenesis and spermatogenesis occurring for 11–12 and 5–6 months, respectively. Timing of spawning of this species was similar to other shallow Acropora spp. in the region. However, colonies had longer gametogenic cycles and less synchronous gamete maturation compared to shallow acroporids with spawning extended over consecutive months. Both the polyp fecundity (number of eggs per polyp) and gonad index (defined as the number of eggs per square centimeter) of A. tenella were lower than most acroporids. Our findings contribute to understanding of the life history of corals on mesophotic reefs and suggest that the reproductive biology of upper mesophotic corals is similar to that of shallow-water corals.     

Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II

One of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment. A promising method to do this is by use of pathogen-specific molecular probes. However, this approach has been little used to date. We constructed, and validated in the laboratory, a fluorochrome-labeled molecular probe specific to Aurantimonas coralicida, the bacterial pathogen of the Caribbean coral disease white plague type II (WPIl). We then used the probe to test field samples of diseased coral tissue for the presence of this pathogen. Probe design was based on a unique subset (25 nucleotides) of the complete l6S rRNA gene sequence derived from a pure culture of the pathogen. The pathogen-specific probe was labeled with the fluorochrome GreenStar* FITC (fluorescein isothiocyanate, GeneDetect Ltd, New Zealand). As a control, we used the universal eubacterial probe EUB 338, labeled with a different fluorochrome (TRITC, tetra-methylrhodamine isothiocyanate). Both probes were applied to laboratory samples of pure cultures of bacteria, and field samples collected from the surface of the disease line of corals exhibiting signs of white plague (types I and II), healthy controls, and corals with an uncharacterized disease ("patchy necrosis"). All samples were analyzed using fluorescence in situ hybridization (FISH). We have determined that the probe is specific to our laboratory culture of the coral pathogen, and does not react with other bacterial species (the eubacterial probe does). The WPII pathogen was detected in association with diseased coral samples collected from coral colonies on reefs of the Bahamas (n= 9 samples) exhibiting signs of both WPI and WPII. Diseased (and healthy) tissue samples (n- 4) from corals exhibiting signs of "patchy necrosis" were also assayed. In this case the results were negative, indicating that the same pathogen is not involved in the two diseases. Incorporation and use of pathogen-specific probes can significantly expand our knowledge of the etiology of coral diseases.     

Reproductive energy investment in corals: scaling with module size

In colonial modular organisms, differences in module size and colony growth patterns among species have the potential to impose varying constraints on reproductive investment. Here, we compare reproductive output among seven morphologically different species of spawning reef corals, and analyse the relationship between reproductive output and module (polyp) size. Reproductive output ranged between 132 and 384 J cm^–2, with lipid constituting the key indicator of energy investment. Lipid decreased by 85–100%, whereas protein and carbohydrate were relatively invariant between pre- and post-spawning tissues in all species, representing 1–15% and <1%, respectively, of the energy investment to reproductive output. The ratio of energy content in reproductive to somatic tissues (gonadosomatic index, GSI) varied among species from 0.20 (Symphyllia recta) to 1.31 (Acropora tenuis), the latter being the highest value reported for any iteroparous marine invertebrate. Surprisingly, small-polyped species (Acropora, Montipora) had 2- to 6-fold higher GSIs than large-polyped ones (Lobophyllia, Symphyllia). Energy equivalents of tissues increased with the 1.50–1.76 power of polyp diameter for somatic tissues and with the 1.42–1.80 power of polyp diameter for reproductive output. In both cases, increases in energy equivalents with polyp diameter were less than the scaling exponent of 3 predicted for an isometric relationship between tissue volume (or mass) and polyp diameter, indicating significant constraints of space, design or physiological energetics with increasing polyp size. We hypothesise that such constraints have played a key role in the evolution of modularity in cnidarians.     

Fecundity and sexual maturity of the coral Siderastrea siderea at high latitude along the Florida Reef Tract,USA

Siderastrea siderea is one of the most abundant corals at high latitude shallow sites along the Florida Reef Tract (25°–27°N). This species is able to tolerate wide seawater temperature fluctuations and sedimentation stress, but its reproductive status at high latitudes and under marginal environmental conditions is poorly understood. The objectives of this study were to evaluate the reproductive potential of S. siderea along a latitudinal gradient (25°–27°N) and to determine if sexual maturity occurs in small (<12.0 cm) S. siderea colonies. Samples of coral tissue were collected in 2007, 2008, and 2009 at three sites along the latitudinal gradient and were processed for histological analysis. Oocyte size, volume, and abundance were used to calculate fecundity. Results showed that fecundity decreased with increasing latitude and that oocyte volume was the major contributing factor to this variation. Mature oocytes were observed in S. siderea colonies at sizes as small as 1.1 cm in diameter. The ability of S. siderea to reach fertility at high latitude areas suggests this species is able to reproduce under marginal environmental conditions; however, reduction in oocyte size could increase local retention of larvae. The presence of mature oocytes in small colonies suggests that stress can reduce somatic growth and shift sexual maturity to smaller colony sizes.     

Live coral predation by fish in Tayrona Nature National Park, Colombian Caribbean

Live coral predation by fish was evaluated in two bays of the Tayrona National Natural Park (Colombia), as a possible biological agent causing coral mortality. Visual censuses were used to identify the most important predator. Predation incidence was determined by examining all colonies present in permanent belt transects (20 x 2 m) in two reef environments (one dominated by Colpophyllia natans and the other one by Montastraea faveolata), for two climatic seasons (rainy and dry seasons). The parrotfish Sparisoma viride was the most important predator due to its biting frequency and bite size. S. viride adults of the initial and terminal phases, removed live tissue and part of the calcareous matrix of M. faveolata, M. annularis, Porites astreoides and C. natans, of which, the last one lost a major amount of tissue per area (3.51 cm2) and volume (3.22 cm3) per bite. A negative exponential tendency (r2=0.94), between coral density and volume removed was found, indicating that the coral density determines the bite's damage. There is no clear relationship between predation incidence and climatic seasons at the sites studied. At Chengue and Gayraca bays, live coral predation is one of the factors contributing to coral tissue loss and could have important consequences on the dynamic of these reefs.     

The Possible Role of Cyanobacterial Filaments in Coral Black Band Disease Pathology

Black band disease (BBD), characterized by a black mat or line that migrates across a coral colony leaving behind it a bare skeleton, is a persistent disease affecting massive corals worldwide. Previous microscopic and molecular examination of this disease in faviid corals from the Gulf of Eilat revealed a number of possible pathogens with the most prominent being a cyanobacterium identified as Pseudoscillatoria coralii. We examined diseased coral colonies using histopathological and molecular methods in order to further assess the possible role of this cyanobacterium, its mode of entry, and pathological effects on the coral host tissues. Affected areas of colonies with BBD were sampled for examination using both light and transmission electron microscopies. Results showed that this dominant cyanobacterium was found on the coral surface, at the coral–skeletal interface, and invading the polyp tissues and gastrovascular cavity. Although tissues surrounding the invasive cyanobacterial filaments did not show gross morphological alterations, microscopic examination revealed that the coral cells surrounding the lesion were dissociated, necrotic, and highly vacuolated. No amoebocytes were evident in the mesoglea of affected tissues suggesting a possible repression of the coral immune response. Morphological and molecular similarity of the previously isolated BBD-associated cyanobacterium P. coralii to the current samples strengthens the premise that this species is involved in the disease in this coral. These results indicate that the cyanobacteria may play a pivotal role in this disease and that the mode of entry may be via ingestion, penetrating the coral via the gastrodermis, as well as through the skeletal–tissue interface.     

Spatial variability in distribution and prevalence of skeletal eroding band and brown band disease in Faafu Atoll,Maldives

Indo-Pacific coral diseases are currently considered one of the 15 globally important threats requiring conservation attention. The coral reefs of the Maldives are experiencing a local decline, with the presence of some coral diseases reported only recently. We investigated the spatial variability in prevalence and distribution of two protozoan diseases, skeletal eroding band (SEB) and brown band disease (BrB), in three islands in the Faafu Atoll: an inhabited island (Magoodhoo), an uninhabited island (Adanga) and a resort island (Filitheyo). Our study revealed a low level of mean disease prevalence for both diseases (<1 %), with Magoodhoo and Adanga being the most affected by BrB and SEB, respectively. However, our preliminary temporal investigations revealed an increment of both coral diseases in Adanga during the last 4 years. Furthermore, we observed different spatial patterns between the two diseases, with SEB positively correlated to dead coral coverage. Finally, Acropora was the most affected coral genus, hosting both coral diseases. Considering that Acropora is the most abundant genus in the archipelago and many other areas in the Indo-Pacific, this finding highlights the need for particular conservation efforts for this genus. These results represent just a first step in the assessment of Maldivian coral disease epidemiology, and more detailed analyses of regional differences in diseases prevalence are needed to further explore their impacts on Maldivian coral reefs.     

Occupation Dynamics and Impacts of Damselfish Territoriality on Recovering Populations of the Threatened Staghorn Coral,Acropora cervicornis

Large-scale coral reef restoration is needed to help recover structure and function of degraded coral reef ecosystems and mitigate continued coral declines. In situ coral propagation and reef restoration efforts have scaled up significantly in past decades, particularly for the threatened Caribbean staghorn coral, Acropora cervicornis, but little is known about the role that native competitors and predators, such as farming damselfishes, have on the success of restoration. Steep declines in A. cervicornis abundance may have concentrated the negative impacts of damselfish algal farming on a much lower number of coral prey/colonies, thus creating a significant threat to the persistence and recovery of depleted coral populations. This is the first study to document the prevalence of resident damselfishes and negative effects of algal lawns on A. cervicornis along the Florida Reef Tract (FRT). Impacts of damselfish lawns on A. cervicornis colonies were more prevalent (21.6% of colonies) than those of other sources of mortality (i.e., disease (1.6%), algal/sponge overgrowth (5.6%), and corallivore predation (7.9%)), and damselfish activities caused the highest levels of tissue mortality (34.6%) among all coral stressors evaluated. The probability of damselfish occupation increased as coral colony size and complexity increased and coral growth rates were significantly lower in colonies with damselfish lawns (15.4 vs. 29.6 cm per year). Reduced growth and mortality of existing A. cervicornis populations may have a significant effect on population dynamics by potentially reducing important genetic diversity and the reproductive potential of depleted populations. On a positive note, however, the presence of resident damselfishes decreased predation by other corallivores, such as Coralliophila and Hermodice, and may offset some negative impacts caused by algal farming. While most negative impacts of damselfishes identified in this study affected large individual colonies and <50% of the A. cervicornis population along the FRT, the remaining wild staghorn population, along with the rapidly increasing restored populations, continue to fulfill important functional roles on coral reefs by providing essential habitat and refuge to other reef organisms. Although the effects of damselfish predation are, and will continue to be, pervasive, successful restoration efforts and strategic coral transplantation designs may help overcome damselfish damage by rapidly increasing A. cervicornis cover and abundance while also providing important information to educate future conservation and management decisions.     

Light microenvironment and single-cell gradients of carbon fixation in tissues of symbiont-bearing corals

Recent coral optics studies have revealed the presence of steep light gradients and optical microniches in tissues of symbiont-bearing corals. Yet, it is unknown whether such resource stratification allows for physiological differences of Symbiodinium within coral tissues. Using a combination of stable isotope labelling and nanoscale secondary ion mass spectrometry, we investigated in hospite carbon fixation of individual Symbiodinium as a function of the local O₂ and light microenvironment within the coral host determined with microsensors. We found that net carbon fixation rates of individual Symbiodinium cells differed on average about sixfold between upper and lower tissue layers of single coral polyps, whereas the light and O₂ microenvironments differed ~15- and 2.5-fold, respectively, indicating differences in light utilisation efficiency along the light microgradient within the coral tissue. Our study suggests that the structure of coral tissues might be conceptually similar to photosynthetic biofilms, where steep physico-chemical gradients define form and function of the local microbial community.The quantity and quality of solar radiation are arguably the most important environmental resources that affect the structure and function of photosynthetic communities in both terrestrial and aquatic environments. Sunlight is of key importance for symbiont-bearing corals, driving the symbiotic interaction between the coral animal and its photosynthetic microalgae of the genus Symbiodinium (Roth, 2014). Light attenuation through the water mass and over the reef matrix has a fundamental role in structuring morphology, function and distribution of corals and their symbiotic algae with depth (Falkowski et al., 1990). Recent studies on the optical properties of corals have shown that light is also a highly stratified resource at the level of individual coral polyps and tissue layers (Wangpraseurt et al., 2014). Steep light gradients exist within the polyp tissues of some corals and light can attenuate by more than an order of magnitude within tissues, that is, comparable to the attenuation that can occur in open oceanic waters between the surface and >25 m of water depth (Kirk, 1994; Wangpraseurt et al., 2012). In this study, we investigated whether such light gradients within coral tissues are correlated with a stratification of Symbiodinium physiology in hospite.We used fibre-optic and electrochemical microsensors together with stable isotopic labelling and nanoscale secondary ion mass spectrometry (NanoSIMS) to estimate single-cell carbon fixation rates across light gradients within coral tissues. We collected several fragments of Favites sp. from the Heron Island reef flat (152°69'' E, 20°299'' S), Great Barrier Reef, Australia. Fragments were cultured under a downwelling photon irradiance (400–700 nm) of ~100 μmol photons per m² per s (12/12 h cycle), in aerated seawater (25 °C, salinity 33). Photosynthesis-irradiance curves for the investigated corals were determined with an imaging pulse amplitude modulated fluorometer (I-PAM, Walz GmbH, Effeltrich, Germany; Ralph et al., 2005). Values for saturating irradiance, E_max, and irradiance at onset of saturation, E_k, were ~350 μmol photons per m² per s and ~160 μmol photons per m² per s, respectively (data not shown). These values are typical for healthy corals kept under moderate irradiance (Ralph et al., 2005). To ensure incubations at irradiance levels where photosynthesis and irradiance correlated linearly, that is, on the linearly increasing part of the P vs I curve, all experiments were performed at ~80 μmol photons per m² per s (12/12 h cycle). Microsensor measurements of scalar irradiance (tip size ~60 μm; Lassen et al., 1992) and O₂ concentration (OX-50, tip size 50 μm, Unisense A/S, Aarhus, Denmark) were performed within the polyp and coenosarc tissues of corals as described previously (Figures 1a and b; Wangpraseurt et al., 2012). After microsensor measurements, corals were incubated with ¹³C-bicarbonate (Supplementary Text S1). NanoSIMS imaging was then applied on coral tissue sections, as described by Pernice et al. (2014) to quantify the assimilation of dissolved inorganic carbon into individual Symbiodinium cells across polyp (oral and aboral) and coenosarc tissues of corals. Briefly, corals were incubated in small aquaria with 2 mm NaH¹³CO₃ in artificial sea water (recipe adapted from Harrison et al., 1980). After 24 h of isotopic incubation, coral fragments were sampled, chemically fixed and processed for NanoSIMS analyses (see Kopp et al., 2013; Pernice et al., 2012, 2014; and Supplementary Text S1,Supplementary Figure S1).Open in a separate windowFigure 1Internal microenvironment and single-cell ¹³C assimilation by Symbiodinium cells within Favites sp. (a) Representative measurement locations indicating connecting tissue (c, coenosarc; white circle) and polyp tissue (p; red circle). Scale bar is 0.5 cm. (b) Schematic diagram of the vertical arrangement of the polyp tissue structure (not drawn to scale). The coral tissue consists of oral and aboral gastrodermal tissues that contain photosymbiont cells (~10 μm in diameter). The two tissue layers are separated by a flexible gastrodermal cavity and the entire mean polyp tissue thickness was 1150 μm (±385 s.d., n=8) as determined by microsensor profiles. The NanoSIMS images (c–e) show the ¹³C/¹²C isotopic ratio for Symbiodinium cells in coenosarc tissue (c), the upper oral polyp tissue (d) and in the lowest layer of aboral polyp tissue (e). Scale bars are 10 μm. The colour scale of the NanoSIMS images is in hue saturation intensity ranging from 220 in blue (which corresponds to natural ¹³C/¹²C isotopic ratio of 0.0110) to 1000 in red (which corresponds to ¹³C/¹²C isotopic ratio of 0.05, ~4.5 times above the natural ¹³C/¹²C isotopic ratio). Quantification of ¹³C enrichment of individual Symbiodinium cells was obtained by selecting regions of interest that were defined in Open_MIMS (http://nrims.harvard.edu/software/openmims) by drawing the contours of the Symbiodinium cells directly on the NanoSIMS images. (f) Mean enrichment measured in Symbiodinium cells by NanoSIMS, in coenosarc tissue (in white, n=33), in upper oral polyp tissue (in grey, n=25), in the lowest layer of polyp tissue (in turquoise, n=17) and in the control treatment (n=20). Bars in the histograms indicate the s.e.m. enrichment quantified for the different whole Symbiodinium cells for each tissue category. Microsensor measurements of (g) scalar irradiance and (h) O₂ performed along depth gradients within the polyp tissue (mean±s.d., n=4). Measurements were averaged for the first 100 μm from the tissue surface (oral) and the last 100 μm from the skeleton (aboral). The oral and aboral depth was defined through gentle touching of the microsensor tip at the surface of the coral tissue and skeleton, respectively.Our combined approach of using NanoSIMS and microsensors within the tissue of corals provides, to the best of our knowledge, the first evidence for physiological differences of individual Symbiodinium cells in hospite in relation to the local microenvironmental conditions across different coral tissue layers, that is, oral vs aboral parts of polyp and coenosarc. Quantitative analysis based on tissue sections from different coral tissue layers showed that mean incorporation of ¹³C-bicarbonate by individual Symbiodinium cells was up to 6.5-fold higher in the upper oral polyp and coenosarc tissues compared with the lowermost layer of polyp tissues (δ¹³C: 1609±147‰, n=25 for Symbiodinium cells in upper oral polyp tissue; 1696±205‰, n=33 for Symbiodinium cells in coenosarc tissue and 246±82‰, n=17 for Symbiodinium cells in the lowest aboral layer of polyp tissue). Although the sample sizes in this study are small and the ¹³C signal is heterogeneous within individual Symbiodinium cells (because of carbon fixation hotspots in specific compartments; Supplementary Figure S2; Kopp et al., 2015), the magnitude of the difference in mean ¹³C incorporation between the aboral part of the polyp and the two other parts of coral tissue was clear and statistically significant (one-way analysis of variance (ANOVA) F_2,75=15.91; P<0.0001; 6.5-fold increase in polyp oral vs aboral polyp tissue, Fischer''s least significant difference (LSD) P<0.0001; 6.9-fold increase in coenosarc vs aboral polyp tissue Fischer''s LSD P<0.0001; and no significant difference between oral polyp vs coenosarc tissue, Fischer''s LSD P=0.718; Figure 1c–f; Supplementary Table S1). The internal microenvironment within the corresponding polyp tissues was highly stratified with respect to light and O₂ (Figures 1g and h). Scalar irradiance decreased about 15-fold from the surface to the bottom of the polyp tissues. Gradients of O₂ were less steep but still significant, with an approximate reduction in O₂ concentration by about 2.5 times (Figure 1; Supplementary Table S2; ANOVA F_1,6= 16.4; P=0.006).These results suggest that coral tissues are vertically stratified systems that affect the physiological activity of their symbionts along a fine-scale microenvironmental gradient. The presence and role of microscale heterogeneity has hitherto largely been ignored in the field of coral symbiosis research, while much is known for other photosynthetic tissues. For instance, for terrestrial plant leaves and for aquatic photosynthetic biofilms, it is known that the photosynthetic unit can adapt to microenvironmental light gradients, where chloroplasts/phototrophs harboured in low-light niches show increased photosynthetic quantum efficiencies at low light levels (Terashima and Hikosaka, 1995; Al-Najjar et al., 2012). Although the steady-state O₂ concentration values reported here are a function of the different metabolic processes of the coral holobiont (that is, Symbiodinium photosynthesis and the combined respiration by the coral host, Symbiodinium and microbes), the NanoSIMS approach allowed us to separate ¹³C fixation of Symbiodinium from the host metabolic activity. Our study provides the first experimental evidence from carbon fixation measurements that Symbiodinium cells can adapt to optical microniches in coral tissues. The 15-fold reduction in irradiance with depth in the coral tissue led only to an ~6.5-fold reduction in net carbon fixation suggesting enhanced light-harvesting efficiency or a reduced P/R ratio for Symbiodinium harboured in aboral tissues. Although such enhanced efficiency under low light often reflects the adaptation of the photosynthetic apparatus (for example, an increase in light-harvesting complexes (Walters, 2005) and reduced cell respiration (Givnish, 1988), it might additionally be the result of physiologically distinct populations or clades of Symbiodinium. Several studies have revealed remarkable genetic and physiological diversities among different Symbiodinium clades (Loram et al., 2007; Stat et al., 2008; Baker et al., 2013; Pernice et al., 2014). Although Favites sp. corals from Southern Great Barrier Reef are generally reported in association with one specific Symbiodinium type (clade C3; Tonk et al., 2013), Symbiodinium diversity within the microenvironment of these common corals could have been overlooked and such physiological diversity could further provide selective advantage to different genotypes in microenvironments within coral tissue. Coral tissues might thus exhibit similar characteristics to photosynthetic biofilms where steep physico-chemical microgradients give rise to different pheno- and ecotypes of phototrophs along those gradients (Musat et al., 2008; Ward et al., 1998).These first experiments were performed under sub-saturating irradiance of ~80 μmol photons per m² per s. Earlier studies showed that the local scalar irradiance in upper vs deeper tissue layers relates to the incident photon irradiance in a linear fashion such that at stressful incident irradiance levels of, for example, 2000 μmol photons per m² per s, light levels in the lowermost polyp tissue layers are ~200 μmol photons per m² per s (Wangpraseurt et al., 2012), still representing optimal conditions for photosynthesis. We thus consider it likely that excess irradiance triggering photoinhibition in oral tissues is unlikely to cause photoinhibition of Symbiodinium in aboral polyp tissues. The internal light field is species specific and in some thin-tissued, branching corals such as Pocillopora damicornis, intra-tissue light attenuation is not very pronounced (Wangpraseurt et al., 2012; Szabó et al., 2014). The ability to harbour Symbiodinium cells in low-light niches might be an important resilience factor for thick-tissued corals, such as massive faviids, during and after coral bleaching. Our study gives first insights to the functional diversity of Symbiodinium along microscale gradients in coral tissue and underscores the importance of considering such heterogeneity in studies linking symbiont diversity and coral physiology responses to environmental stress factors.     

Implications of coral harvest and transplantation on reefs in northwestern Dominica

In June, 2002, the government of Dominica requested assistance in evaluating the coral culture and transplantation activities being undertaken by Oceanographic Institute of Dominica (OID), a coral farm culturing both western Atlantic and Indo-Pacific corals for restoration and commercial sales. We assessed the culture facilities of OID, the condition of reefs, potential impacts of coral collection and benefits of coral transplantation. Coral reefs (9 reefs, 3-20 m depth) were characterized by 35 species of scleractinian corals and a live coral cover of 8-35%. Early colonizing, brooders such as Porites astreoides (14.8% of all corals), P. porites (14.8%), Meandrina meandrites (14.7%) and Agaricia agaricites (9.1%) were the most abundant corals, but colonies were mostly small (mean = 25 cm diameter). Montastraea annularis (complex) was the other dominant taxa (20.8% of all corals) and colonies were larger (mean = 70 cm). Corals (pooled species) were missing an average of 20% of their tissue, with a mean of 1.4% recent mortality. Coral diseases affected 6.4% of all colonies, with the highest prevalence at Cabrits West (11.0%), Douglas Bay (12.2%) and Coconut Outer reef (20.7%). White plague and yellow band disease were causing the greatest loss of tissue, especially among M. annularis (complex), with localized impacts from corallivores, overgrowth by macroalgae, storm damage and sedimentation. While the reefs appeared to be undergoing substantial decline, restoration efforts by OlD were unlikely to promote recovery. No Pacific species were identified at OID restoration sites, yet species chosen for transplantation with highest survival included short-lived brooders (Agaricia and Porites) that were abundant in restoration sites, as well as non-reef builders (Palythoa and Erythropodium) that monopolize substrates and overgrow corals. The species of highest value for restoration (massive broadcast spawners) showed low survivorship and unrestored populations of these species were most affected by biotic stressors and human impacts, all of which need to be addressed to enhance survival of outplants. Problems with culture practices at OID, such as high water temperature, adequate light levels and persistent overgrowth by macroalgae could be addressed through simple modifications. Nevertheless, coral disease and other stressors are of major concern to the most important reef builders, as these species are less amenable to restoration, collection could threaten their survival and losses require decades to centuries to replace.