Development of bacterial biofilms on artificial corals in comparison to surface-associated microbes of hard corals

Numerous studies have demonstrated the differences in bacterial communities associated with corals versus those in their surrounding environment. However, these environmental samples often represent vastly different microbial micro-environments with few studies having looked at the settlement and growth of bacteria on surfaces similar to corals. As a result, it is difficult to determine which bacteria are associated specifically with coral tissue surfaces. In this study, early stages of passive settlement from the water column to artificial coral surfaces (formation of a biofilm) were assessed. Changes in bacterial diversity (16S rRNA gene), were studied on artificially created resin nubbins that were modelled from the skeleton of the reef building coral Acropora muricata. These models were dip-coated in sterile agar, mounted in situ on the reef and followed over time to monitor bacterial community succession. The bacterial community forming the biofilms remained significantly different (R = 0.864 p<0.05) from that of the water column and from the surface mucus layer (SML) of the coral at all times from 30 min to 96 h. The water column was dominated by members of the α-proteobacteria, the developed community on the biofilms dominated by γ-proteobacteria, whereas that within the SML was composed of a more diverse array of groups. Bacterial communities present within the SML do not appear to arise from passive settlement from the water column, but instead appear to have become established through a selection process. This selection process was shown to be dependent on some aspects of the physico-chemical structure of the settlement surface, since agar-coated slides showed distinct communities to coral-shaped surfaces. However, no significant differences were found between different surface coatings, including plain agar and agar enhanced with coral mucus exudates. Therefore future work should consider physico-chemical surface properties as factors governing change in microbial diversity.     

Observations of a new source of coral mortality along the Kenyan coast

In early 2002 coral mortality occurred along 600 km of coastline from Tanzania to Kenya. Astreopora, Echinopora , and Montipora species were severely affected, with Montipora being nearly eliminated from Kenyan reefs. Acropora , Platygyra , Goniopora , and massive Porites were also affected; however, Porites and Goniopora rarely died and often recovered, whereas death for most other species occurred within 2 weeks. In Echinopora and Montipora , a dull ashy tissue color and brittle skeletons characterized the early stages of this event with a mucus layer on the tissue surface in intermediate stages. Mucus and embedded debris then disappeared and surfaces were left covered in a white calcareous dust that sometimes capped a black layer. Astreopora tissues became dull and pale, and seldom produced mucus; eventually the skeleton became bare and white. Either a colorless translucent or brownish thin margin of tissue was visible between living tissue and bare skeleton, depending on species. Scanning electron micrographs of affected corals revealed the presence of fungi. Histology and staining showed that the fungi were mostly in the three genera that died from the syndrome and it may be that fungi invaded and killed corals weakened by another unidentified pathogen.     

Bacteria Associated with the Bleached and Cave Coral Oculina patagonica

The relative abundance of bacteria in the mucus and tissues of Oculina patagonica taken from bleached and cave (azooxanthellae) corals was determined by analyses of the 16S rRNA genes from cloned libraries of extracted DNA and from isolated colonies. The results were compared to previously published data on healthy O. patagonica. The bacterial community of bleached, cave, and healthy corals were completely different from each other. A tight cluster (>99.5% identity) of bacteria, showing 100% identity to Acinetobacter species, dominated bleached corals, comprising 25% of the 316 clones sequenced. The dominant bacterial cluster found in cave corals, representing 29% of the 97 clones sequenced, showed 98% identity to an uncultured bacterium from the Great Barrier Reef. Vibrio splendidus was the most dominant species in healthy O. patagonica. The culturable bacteria represented 0.1–1.0% of the total bacteria (SYBR Gold staining) of the corals. The most abundant culturable bacteria in bleached, cave, and healthy corals were clusters that most closely matched Microbulbifer sp., an α-proteobacterium previously isolated from healthy corals and an α-protobacterium (AB026194), respectively. Three generalizations emerge from this study on O. patagonica: (1) More bacteria are associated with coral tissue than mucus; (2) tissue and mucus populations are different; (3) bacterial populations associated with corals change dramatically when corals lack their symbiotic zooxanthellae, either as a result of the bleaching disease or when growing in the absence of light.     

Bacterial communities associated with healthy and Acropora white syndrome-affected corals from American Samoa

Acropora white syndrome (AWS) is characterized by rapid tissue loss revealing the white underlying skeleton and affects corals worldwide; however, reports of causal agents are conflicting. Samples were collected from healthy and diseased corals and seawater around American Samoa and bacteria associated with AWS characterized using both culture-dependent and culture-independent methods, from coral mucus and tissue slurries, respectively. Bacterial 16S rRNA gene clone libraries derived from coral tissue were dominated by the Gammaproteobacteria, and Jaccard's distances calculated between the clone libraries showed that those from diseased corals were more similar to each other than to those from healthy corals. 16S rRNA genes from 78 culturable coral mucus isolates also revealed a distinct partitioning of bacterial genera into healthy and diseased corals. Isolates identified as Vibrionaceae were further characterized by multilocus sequence typing, revealing that whilst several Vibrio spp. were found to be associated with AWS lesions, a recently described species, Vibrio owensii, was prevalent amongst cultured Vibrio isolates. Unaffected tissues from corals with AWS had a different microbiota than normal Acropora as found by others. Determining whether a microbial shift occurs prior to disease outbreaks will be a useful avenue of pursuit and could be helpful in detecting prodromal signs of coral disease prior to manifestation of lesions.     

Heavy metals distribution in the coral reef ecosystems of the Northern Red Sea

Concentrations of seven heavy metals (Cu, Zn, Pb, Cd, Ni, Co and Fe) were measured in the seawater, sediments, common scleractinian reef-building corals and soft corals (Octocorallia : Alcyonacea) at seven reef sites in the Northern Red Sea: I (Hurghada), II (Ras Za’farana), III (El-Ain Al-Sukhna), IV (El-Tur), V (Sha’b Rashdan), VI (Sharm El-Sheikh) and VII (Dahab). Levels of heavy metals were considerably elevated in seawater, sediments and corals collected from reef sites exposed to increased environmental contamination, as a result of diversified natural and anthropogenic inputs. Soft corals of genera Lithophyton, Sarcophyton and Sinularia showed higher concentrations of Zn, Pb, Cd and Ni than hard coral genera Acropora and Stylophora. Soft coral Sarcophyton trocheliophorum collected from El Ain Al-Suhkna (Gulf of Suez) had greater concentration of Cu, followed by hard corals Acropora pharaonis and Acropora hemprichi. The elevated levels of Zn, Cd and Ni were reported in the dry tissue of soft coral Sinularia spp. On the other hand, the soft coral Lithophyton arboreum displayed the highest concentration of Pb at Sha’b Rashdan (Gulf of Suez) and elevated concentration of Zn at Sharm El-Sheikh. Sediments showed significantly higher concentration of Fe than corals. The higher levels of Fe in hard corals than soft corals reflected the incorporation of Fe into the aragonite and the chelation with the organic matrix of the skeleton. The greater abundance of soft corals in metal-contaminated reef sites and the elevated levels of metals in their tissue suggesting that the soft corals could develop a tolerance mechanism to relatively high concentrations of metals. Although the effects of heavy metals on reef corals were not isolated from the possible effects of other stresses, the percentage cover of dead corals were significantly higher as the concentrations of heavy metals increased.     

Response of triassic reef coral communities to sea-level fluctuations, storms and sedimentation: Evidence from a spectacular outcrop (Adnet, Austria)

Summary The Upper Rhaetian coral limestone of Adnet, southeast of Salzburg Austria has been repeatedly referred to as one of the most spectacular examples of an ancient ‘autochthonous’ coral reef structure. The ‘Tropfbruch’ quarry is probably the best outcrop for interpreting the distributional patterns of biotic successions and communities of a late Triassic patch reef. Our study is based on the interpretation of a) outcrop photographs, b) reef maps resulting from quadrat transects, and c) the analysis of quantitative data describing the distribution and frequency of reef organisms and sediment. A new methodological approach (combination of reef mapping and photo-transects) is used to obtain quantitative field data which can be compared in greater detail with data from modern coral reefs investigated by corresponding quantitative surveys. Three unconformities and three well-defined ‘reef growth stages’ reflecting the vertical and lateral development of the reef structure were differrentiated using transects: Stage 1, representing the reef growth optimum, is characterized by laterally differentiated coral reef knobs with corals in growth position. Criteria supporting this interpretation are the extraordinary size of the corals, their preservation in situ and the great thickness of this interval. The massive coralPamiroseris grew under higher energy conditions at the rim of the reef knob, whereas branchingRetiophyllia colonies preferred less agitated water in the center. Vertical changes are reflected by an increase in frequency of the dasycladacean algaDiplopora adnetensis and by the decreasing size ofRetiophyllia. These sedimentological and biological criteria together with the unconformity above indicate a fall in the sea level as a major control mechanism. Stage 2, separated from stage 1 by an unconformity caused by partial subaerial exposure and karstification, is characterized by vertically stacked coral successions with diverse reef debris. Facies heterogeneity is reflected by differences in the diversity, taphonomy and packing density of reef-building organisms as well as by differences in sediment input from the platform. Water depths and accommodation space were lower, therefore minor sea level fluctuations had a stronger effect on the biotic composition. The high percentage of coral debris and corals reworked by storms and the increase in the input of platform sediment led to a reduction of reef growth. Stage 3, again separated at the base by an unconformity associated with karstification, is characterized by bioclastic sediments with isolated reefbuilders forming a level-bottom community. The distribution of different coral morphotypes suggests that sea level fluctuations were not the only controlling factor. Variations in the substrate were caused by differences in the input of platform sediment. The three-step development seen in Adnet documents the response of low-diverse coral associations to variations caused by small-scale sea level changes, storm activity and sedimentation. The vertical changes in reef community structures correspond to a sequence of ‘allogenic replacements’. The Adnet reef structure should not be regarded as a general model of Alpine Upper Rhaetian reefs, because of the particular setting of the patch reef. Only the ‘capping beds’ of the Upper Rhaetian Reef Limestone of the Steinplatte exhibit criteria similar to Adnet. Potential modern analogues of features seen in the coral communities of Adnet are the internal structure of theRetiophyllia thickets, the key role of branching corals within the communities, the scattered distribution and low and even diversity of corals subsequent to breaks in settlement, segration patterns of corals indicating ‘contact avoidance’, toppling of large coral colonies by intensive boring, and decreasing coral coverage from deeper and sheltered settings to more shallower water depths.     

PhyloChip™ microarray comparison of sampling methods used for coral microbial ecology

Interest in coral microbial ecology has been increasing steadily over the last decade, yet standardized methods of sample collection still have not been defined. Two methods were compared for their ability to sample coral-associated microbial communities: tissue punches and foam swabs, the latter being less invasive and preferred by reef managers. Four colonies of star coral, Montastraea annularis, were sampled in the Dry Tortugas National Park (two healthy and two with white plague disease). The PhyloChip™ G3 microarray was used to assess microbial community structure of amplified 16S rRNA gene sequences. Samples clustered based on methodology rather than coral colony. Punch samples from healthy and diseased corals were distinct. All swab samples clustered closely together with the seawater control and did not group according to the health state of the corals. Although more microbial taxa were detected by the swab method, there is a much larger overlap between the water control and swab samples than punch samples, suggesting some of the additional diversity is due to contamination from water absorbed by the swab. While swabs are useful for noninvasive studies of the coral surface mucus layer, these results show that they are not optimal for studies of coral disease.     

Spatial Homogeneity of Bacterial Communities Associated with the Surface Mucus Layer of the Reef-Building Coral Acropora palmata

Coral surface mucus layer (SML) microbiota are critical components of the coral holobiont and play important roles in nutrient cycling and defense against pathogens. We sequenced 16S rRNA amplicons to examine the structure of the SML microbiome within and between colonies of the threatened Caribbean reef-building coral Acropora palmata in the Florida Keys. Samples were taken from three spatially distinct colony regions—uppermost (high irradiance), underside (low irradiance), and the colony base—representing microhabitats that vary in irradiance and water flow. Phylogenetic diversity (PD) values of coral SML bacteria communities were greater than surrounding seawater and lower than adjacent sediment. Bacterial diversity and community composition was consistent among the three microhabitats. Cyanobacteria, Bacteroidetes, Alphaproteobacteria, and Proteobacteria, respectively were the most abundant phyla represented in the samples. This is the first time spatial variability of the surface mucus layer of A. palmata has been studied. Homogeneity in the microbiome of A. palmata contrasts with SML heterogeneity found in other Caribbean corals. These findings suggest that, during non-stressful conditions, host regulation of SML microbiota may override diverse physiochemical influences induced by the topographical complexity of A. palmata. Documenting the spatial distribution of SML microbes is essential to understanding the functional roles these microorganisms play in coral health and adaptability to environmental perturbations.     

Regulation of Bacterial Communities Through Antimicrobial Activity by the Coral Holobiont

Interactions between corals and associated bacteria and amongst these bacterial groups are likely to play a key role in coral health. However, the complexity of these interactions is poorly understood. We investigated the functional role of specific coral-associated bacteria in maintaining microbial communities on the coral Acropora millepora (Ehrenberg 1834) and the ability of coral mucus to support or inhibit bacterial growth. Culture-independent techniques were used to assess bacterial community structures whilst bacterial culture was employed to assess intra- and inter-specific antimicrobial activities of bacteria. Members of Pseudoalteromonas and ribotypes closely related to Vibrio coralliilyticus displayed potent antimicrobial activity against a range of other cultured isolates and grew readily on detached coral mucus. Although such bacterial ribotypes would be expected to have a competitive advantage, they were rare or absent on intact and healthy coral colonies growing in situ (analysed using denaturing gradient gel electrophoresis and 16S rRNA gene sequencing). The most abundant bacterial ribotypes found on healthy corals were Gammaproteobacteria, previously defined as type A coral associates. Our results indicate that this group of bacteria and specific members of the Alphaproteobacteria described here as ‘type B associates’ may be important functional groups for coral health. We suggest that bacterial communities on coral are kept in check by a combination of host-derived and microbial interactions and that the type A associates in particular may play a key role in maintaining stability of microbial communities on healthy coral colonies.     

Bacterial decomposition of coral mucus as evaluated by long-term and quantitative observation

Coral mucus released from Acropora formosa and Montipora digitata was incubated with bacteria under dark conditions for 1 year to evaluate the quantitative degradability. All the mucus samples showed a similar decomposition pattern: about 80% of total organic carbon (TOC) in the mucus was mineralized within 1 month, while some mucus was slowly decomposed over the 1 year. Regression analysis using an exponential curve considering three degradability pools (labile, semilabile, and refractory) fitted the changes of the TOC concentrations very well (r ² > 0.99). Compiling the data on the two coral species, the labile organic C in the coral mucus had mineralization rates of 10–18% d⁻¹ and accounted for 79–87% of the initial TOC in the mucus. Semilabile organic C had mineralization rates of 0.3−1.6% d⁻¹ and accounted for 11−18% of the initial TOC. Refractory organic C accounted for 6% at most. These results suggest that not all coral mucus is rapidly decomposed by bacteria but some mucus remains as semilabile and refractory organic matter for several months.     

Symbiotic crabs maintain coral health by clearing sediments

Stony corals are the foundation of coral reef ecosystems and form associations with other reef species. Many of these associations may be ecologically important and play a role in maintaining the health and diversity of reef systems, rendering it critical to understand the influence of symbiotic organisms in mediating responses to perturbation. This study demonstrates the importance of an association with trapeziid crabs in reducing adverse effects of sediments deposited on corals. In a field experiment, mortality rates of two species of branching corals were significantly lowered by the presence of crabs. All outplanted corals with crabs survived whereas 45–80% of corals without crabs died within a month. For surviving corals that lacked crabs, growth was slower and tissue bleaching and sediment load were higher. Laboratory experiments revealed that corals with crabs shed substantially more of the sediments deposited on coral surfaces, but also that crabs were most effective at removing grain sizes that were most damaging to coral tissues. The mechanism underlying this symbiotic relationship has not been recognized previously, and its role in maintaining coral health is likely to become even more critical as reefs worldwide experience increasing sedimentation.     

δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N in tissue of coral polyps and epilithic algae inhabiting damaged coral colonies under the influence of different light intensities

Studies were performed of the carbon and nitrogen stable isotope (δ¹³C and δ¹⁵N) composition (δ¹³C and δ¹⁵N) of the corals Porites cylindrica and P. lutea (5 years after damaging the colonies by the bleaching events) and of epilithic algae settled onto damaged areas of coral colonies. Coral polyps and three epilithic algal communities (‘red algal turf, green algal turf and red calcified crusts’) were sampled along the boundary between communities of coral polyps and algal colonizers from differently illuminated habitats from 2 to 90% of incident surface photosynthetically active radiation (PAR₀). It was found that communities with a predominance of red algae significantly differed from communities with a predominance of green algae in δ¹³C but not in δ¹⁵N values. An influence of habitat irradiance was found only for communities of coral polyps for δ¹³C and δ¹⁵N values: under bright light (70–90% PAR₀) polyp tissues of both coral species were significantly enriched in heavy carbon isotopes and insignificantly in nitrogen isotopes (δ¹³C values difference ~4‰) relative to tissues of corals under lower light 15–50% PAR₀. On the basis of these results we assumed that differences in light intensities in the habitat ranging from 15 to 90% PAR₀ do not influence on accessibility of the main carbon and nitrogen sources for corals and algae, and exchange by these elements between organisms. We also assumed that the relative enrichment in the heavy carbon isotopes of coral tissues in high light is a result of decreased isotope fractionation (or the absence of fractionation in photosynthesis of their zooxanthellae).     

Do fluctuating temperature environments elevate coral thermal tolerance?

In reef corals, much research has focused on the capacity of corals to acclimatize and/or adapt to different thermal environments, but the majority of work has focused on distinctions in mean temperature. Across small spatial scales, distinctions in daily temperature variation are common, but the role of such environmental variation in setting coral thermal tolerances has received little attention. Here, we take advantage of back-reef pools in American Samoa that differ in thermal variation to investigate the effects of thermally fluctuating environments on coral thermal tolerance. We experimentally heat-stressed Acropora hyacinthus from a thermally moderate lagoon pool (temp range 26.5–33.3°C) and from a more thermally variable pool that naturally experiences 2–3 h high temperature events during summer low tides (temp range 25.0–35°C). We compared mortality and photosystem II photochemical efficiency of colony fragments exposed to ambient temperatures (median: 28.0°C) or elevated temperatures (median: 31.5°C). In the heated treatment, moderate pool corals showed nearly 50% mortality whether they hosted heat-sensitive (49.2 ± 6.5% SE; C2) or heat-resistant (47.0 ± 11.2% SE; D) symbionts. However, variable pool corals, all of which hosted heat-resistant symbionts, survived well, showing low mortalities (16.6 ± 8.8% SE) statistically indistinguishable from controls held at ambient temperatures (5.1–8.3 ± 3.3–8.3% SE). Similarly, moderate pool corals hosting heat-sensitive algae showed rapid rates of decline in algal photosystem II photochemical efficiency in the elevated temperature treatment (slope = −0.04 day⁻¹ ± 0.007 SE); moderate pool corals hosting heat-resistant algae showed intermediate levels of decline (slope = −0.039 day⁻¹ ± 0.007 SE); and variable pool corals hosting heat-resistant algae showed the least decline (slope = −0.028 day⁻¹ ± 0.004 SE). High gene flow among pools suggests that these differences probably reflect coral acclimatization not local genetic adaptation. Our results suggest that previous exposure to an environmentally variable microhabitat adds substantially to coral–algal thermal tolerance, beyond that provided by heat-resistant symbionts alone.     

Comprehensive characterization of skeletal tissue growth anomalies of the finger coral Porites compressa

The scleractinian finger coral Porites compressa has been documented to develop raised growth anomalies of unknown origin, commonly referred to as “tumors”. These skeletal tissue anomalies (STAs) are circumscribed nodule-like areas of enlarged skeleton and tissue with fewer polyps and zooxanthellae than adjacent tissue. A field survey of the STA prevalence in Oahu, Kaneohe Bay, Hawaii, was complemented by laboratory analysis to reveal biochemical, histological and skeletal differences between anomalous and reference tissue. MutY, Hsp90a1, GRP75 and metallothionein, proteins known to be up-regulated in hyperplastic tissues, were over expressed in the STAs compared to adjacent normal-appearing and reference tissues. Histological analysis was further accompanied by elemental and micro-structural analyses of skeleton. Anomalous skeleton was of similar aragonite composition to adjacent skeleton but more porous as evidenced by an increased rate of vertical extension without thickening. Polyp structure was retained throughout the lesion, but abnormal polyps were hypertrophied, with increased mass of aboral tissue lining the skeleton, and thickened areas of skeletogenic calicoblastic epithelium along the basal floor. The latter were highly metabolically active and infiltrated with chromophore cells. These observations qualify the STAs as hyperplasia and are the first report in poritid corals of chromophore infiltration processes in active calicoblastic epithelium areas.     

Phage Therapy of Coral White Plague Disease: Properties of Phage BA3

The bacteriophage BA3 multiplies in and lyses the coral pathogen Thalassomonas loyana. The complete genome of phage BA3 was sequenced; it contains 47 open reading frames with a 40.9% G + C content. Phage BA3 adsorbed to its starved host in seawater with a k = 1.0 × 10⁻⁶ phage ml⁻¹ min⁻¹. Phage therapy of coral disease in aquarium experiments was successful when the phage was added at the same time as the pathogen or 1 day later, but failed to protect the coral when added 2 days after bacterial infection. When the phages were added 1 day after coral infection, the phage titer increased about 100-fold and remained present in the aquarium water throughout the 37-day experiment. At the end of the experiment, the concentration of phages associated with the corals was 2.5 ± 0.5 × 10⁴ per cm² of coral surface. Corals that were infected with the pathogen and treated with phage did not transmit the disease to healthy corals.     

Effects of increased pCO2 on zinc uptake and calcification in the tropical coral Stylophora pistillata

Zinc (Zn) is an essential element for corals. We investigated the effects of ocean acidification on zinc incorporation, photosynthesis, and gross calcification in the scleractinian coral Stylophora pistillata. Colonies were maintained at normal pH_T (8.1) and at two low-pH conditions (7.8 and 7.5) for 5 weeks. Corals were exposed to ⁶⁵Zn dissolved in seawater to assess uptake rates. After 5 weeks, corals raised at pH_T (8.1) exhibited higher ⁶⁵Zn activity in the coral tissue and skeleton, compared with corals raised at a lower pH. Photosynthesis, photosynthetic efficiency, and gross calcification, measured by ⁴⁵Ca incorporation, were however unchanged even at the lowest pH.     

Coral mortality,recovery and reef degradation at Mexico Rocks Patch Reef Complex,Northern Belize,Central America: 1995–1997

The 1995 coral bleaching event in the western Caribbean was the first reported episode that significantly affected the Belize barrier and lagoonal patch reefs. Bleaching was attributed to a 2 mo period of warm water temperatures above 30°C. Near Ambergris Caye, barrier and patch reefs experienced up to 50% bleaching. At Mexico Rocks patch reef complex, the bleaching resulted in changes in reef health, community, and physical structure. Prior to the hyperthermal episode, patch reef surface area consisted of 47% healthy framework coral coverage, 12% secondarily colonized biotic coverage, 35% dead coral surfaces that were degraded by biological activity and physical erosion, and 6%cavities. six months after bleaching, most corals had regained their color, but, owing to coral mortality, areas of surface degradation had increased to an average 49% (p=0.029 based on Kruskal–Wallis analyses). Eighteen months after bleaching, degraded surface areas expanded to 53% (p=0.0366). Although re-coloring indicates rapid recovery for surviving corals, the persistence in dead coral surfaces suggests that reef skeletal structure recovery lags behind that of individual corals. Initial results of framework measurements indicate that bleaching events may result in an ‘imbalance’ in the carbonate production rate of coral reefs and produce mass wasting of the skeletal structure. Remapping of reef skeletal structure should establish quantitative measures for the long-term effects of bleaching on patch reef frameworks.     

High levels of acrylate in the Great Barrier Reef coral Acropora millepora

High concentrations of acrylate, 542–683 μmol g⁻¹ of the non-skeletal dry mass (DM), were measured in the Great Barrier Reef coral, Acropora millepora, using quantitative nuclear magnetic resonance spectroscopy (qNMR). As the amount of NaCl salt in the samples was substantial but variable, the total carbon (TC) in the coral extracts was determined, and the carbon due to acrylate found to represent 13–15% of the TC present in the total organic extracts (TOE). Acrylate, a C₃ compound, is thus a substantial carbon source in the coral holobiont and is known to be derived from dimethylsulfoniopropionate (DMSP), which has previously been found in corals and other organisms that harbor Symbiodinium spp. The reason for such high levels of acrylate in the corals is unknown; possible functions include antimicrobial and/or antioxidant roles, as well as playing a role in the structuring of the healthy resident coral bacteria.     

Corals form characteristic associations with symbiotic nitrogen-fixing bacteria

The complex symbiotic relationship between corals and their dinoflagellate partner Symbiodinium is believed to be sustained through close associations with mutualistic bacterial communities, though little is known about coral associations with bacterial groups able to fix nitrogen (diazotrophs). In this study, we investigated the diversity of diazotrophic bacterial communities associated with three common coral species (Acropora millepora, Acropora muricata, and Pocillopora damicormis) from three midshelf locations of the Great Barrier Reef (GBR) by profiling the conserved subunit of the nifH gene, which encodes the dinitrogenase iron protein. Comparisons of diazotrophic community diversity among coral tissue and mucus microenvironments and the surrounding seawater revealed that corals harbor diverse nifH phylotypes that differ between tissue and mucus microhabitats. Coral mucus nifH sequences displayed high heterogeneity, and many bacterial groups overlapped with those found in seawater. Moreover, coral mucus diazotrophs were specific neither to coral species nor to reef location, reflecting the ephemeral nature of coral mucus. In contrast, the dominant diazotrophic bacteria in tissue samples differed among coral species, with differences remaining consistent at all three reefs, indicating that coral-diazotroph associations are species specific. Notably, dominant diazotrophs for all coral species were closely related to the bacterial group rhizobia, which represented 71% of the total sequences retrieved from tissue samples. The species specificity of coral-diazotroph associations further supports the coral holobiont model that bacterial groups associated with corals are conserved. Our results suggest that, as in terrestrial plants, rhizobia have developed a mutualistic relationship with corals and may contribute fixed nitrogen to Symbiodinium.     

A mechanism of transmission and factors affecting coral susceptibility to <Emphasis Type="Italic">Halofolliculina</Emphasis> sp. infection

Anecdotal evidence collected since 2004 suggests that infections caused by ciliates in the genus Halofolliculina may be related to coral mortality in more than 25 scleractinian species in the Caribbean. However, the relationship between the presence of ciliates and coral mortality has not yet been firmly established. Field and laboratory manipulations were used to test if ciliate infections harm corals, if ciliates are able to infect healthy colonies, and if coral susceptibility to ciliate infection depends on temperature, depth, distance to an infected colony, and the presence of injuries. Ciliate infections were always characterized by a visually detectable front of ciliates located on recently exposed coral skeletons. These infections altered the normal structure of the colony by causing tissue mortality (0.8 ± 0.95 cm month⁻¹, mean ± SD) and by delaying or preventing recovery from injuries. Under laboratory conditions, ciliates transmitted directly and horizontally from infected to healthy hosts, and coral susceptibility to ciliate infections increased with the presence of injuries. After invasion, the ciliate population grew, rapidly and after 8 d, produced tissue mortality on 32% of newly infected hosts. Thus, our results support the existence of a new Caribbean coral syndrome that is associated with tissue mortality, is infectious, and transmits directly and horizontally. Even though the role of ciliates in the development of lesions on coral tissues remains unclear, their presence is by far the most conspicuous sign of this syndrome; thus, we propose to name this condition Caribbean ciliate infection (CCI). Communicated by Biology Editor Dr Michael Lesser