Molecular Investigation of Recurrent Streptococcus iniae Epizootics Affecting Coral Reef Fish on an Oceanic Island Suggests at Least Two Distinct Emergence Events.

Streptococcus iniae is an emerging zoonotic pathogen of increasing concern for aquaculture and has caused several epizootics in reef fishes from the Caribbean, the Red Sea and the Indian Ocean. To study the population structure, introduction pathways and evolution of S. iniae over recurring epizootics on Reunion Island, we developed and validated a Multi Locus Sequence Typing (MLST) panel using genomic data obtained from 89 isolates sampled during epizootics occurring over the past 40years in Australia, Asia, the United States, Israel and Reunion Island. We selected eight housekeeping loci, which resulted in the greatest variation across the main S. iniae phylogenetic clades highlighted by the whole genomic dataset. We then applied the developed MLST to investigate the origin of S. iniae responsible for four epizootics on Reunion Island, first in inland aquaculture and then on the reefs from 1996 to 2014. Results suggest at least two independent S. iniae emergence events occurred on the island. Molecular data support that the first epizootic resulted from an introduction, with inland freshwater aquaculture facilities acting as a stepping-stone. Such an event may have been facilitated by the ecological flexibility of S. iniae, able to survive in both fresh and marine waters and the ability of the pathogen to infect multiple host species. By contrast, the second epizootic was associated with a distinct ST of cosmopolitan distribution that may have emerged as a result of environment disturbance. This novel tool will be effective at investigating recurrent epizootics occurring within a given environment or country that is despite the fact that S. iniae appears to have low genetic diversity within its lineage.

Insights into the Cultured Bacterial Fraction of Corals.

Bacteria associated with coral hosts are diverse and abundant, with recent studies suggesting involvement of these symbionts in host resilience to anthropogenic stress. Despite their putative importance, the work dedicated to culturing coral-associated bacteria has received little attention. Combining published and unpublished data, here we report a comprehensive overview of the diversity and function of culturable bacteria isolated from corals originating from tropical, temperate, and cold-water habitats. A total of 3,055 isolates from 52 studies were considered by our metasurvey. Of these, 1,045 had full-length 16S rRNA gene sequences, spanning 138 formally described and 12 putatively novel bacterial genera across the Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria phyla. We performed comparative genomic analysis using the available genomes of 74 strains and identified potential signatures of beneficial bacterium-coral symbioses among the strains. Our analysis revealed >400 biosynthetic gene clusters that underlie the biosynthesis of antioxidant, antimicrobial, cytotoxic, and other secondary metabolites. Moreover, we uncovered genomic features-not previously described for coral-bacterium symbioses-potentially involved in host colonization and host-symbiont recognition, antiviral defense mechanisms, and/or integrated metabolic interactions, which we suggest as novel targets for the screening of coral probiotics. Our results highlight the importance of bacterial cultures to elucidate coral holobiont functioning and guide the selection of probiotic candidates to promote coral resilience and improve holistic and customized reef restoration and rehabilitation efforts. IMPORTANCE Our paper is the first study to synthesize currently available but decentralized data of cultured microbes associated with corals. We were able to collate 3,055 isolates across a number of published studies and unpublished collections from various laboratories and researchers around the world. This equated to 1,045 individual isolates which had full-length 16S rRNA gene sequences, after filtering of the original 3,055. We also explored which of these had genomes available. Originally, only 36 were available, and as part of this study, we added a further 38-equating to 74 in total. From this, we investigated potential genetic signatures that may facilitate a host-associated lifestyle. Further, such a resource is an important step in the selection of probiotic candidates, which are being investigated for promoting coral resilience and potentially applied as a novel strategy in reef restoration and rehabilitation efforts. In the spirit of open access, we have ensured this collection is available to the wider research community through the web site http://isolates.reefgenomics.org/ with the hope many scientists across the globe will ask for access to these cultures for future studies.

Estimations of densities of marine litter on the fringing reefs of Mayotte (France - South Western Indian Ocean) - impacts on coral communities.

The present study was carried out at 22 stations located on the fringing reefs of the island of Mayotte inside the perimeter of the Marine Natural Park of Mayotte (PNMM). The average assessment of the number of samples of marine litter (ML) collected at these stations was 3.9 ± 1.3 ML per 500 m2 in winter and 3.8 ± 1.1 ML per 500 m2 in summer. Plastic ML was dominant, representing approximately 92% of the litter collected at the stations for the two seasons. The majority was plastic ML from fishing gear (lines, nets etc.). Station 18 presented remarkable average densities of 39 ± 4.2 ML per 500 m2 for the two seasons and differed from the other stations in that the ML came solely from the catchment areas, with a majority of aluminium ML (56%). Approximately half the coral colonies coming into contact with ML presented an impact dominated by the category of "Broken or abraded colonies", with 25% of the colonies being impacted. The colonies most severely impacted by ML were branch or table corals, with the greatest majority for the present study being of the genus Acropora.

What can South African reefs tell us about the future of high-latitude coral systems?

Coral communities are found at high latitude on the East Coast subtropical reefs of South Africa. They are biodiverse, economically important, and afforded World Heritage Site status in the iSimangaliso Wetland Park where some are subjected to recreational use. While the Park's unique coral reefs have, to date, suffered little bleaching from climate change, they are susceptible to the phenomenon and provide a natural laboratory for the study of its effects at high latitude. This review covers recent advances in the regional oceanography; coral community dynamics and the underpinning reef processes, including minor bleaching events; the incidence of coral disease; and coral genetic connectivity. The effects of human activity (SCUBA diving, recreational fishing, pesticide use) were assessed, as well as the nursery benefits of Acropora austera, a coral which provides the reefs with much structure and is vulnerable to damage and climate change. The reefs were valued in terms of human use as well as services such as sediment generation and retention. The results have provided valuable information on relatively pristine, high-latitude reefs, their socio-economic benefits, and the anticipated effects of climate change.

Characterisation of an atypical manifestation of black band disease on Porites lutea in the Western Indian Ocean.

Recent surveys conducted on Reunion Island coral reefs revealed an atypical manifestation of black band disease on the main framework building coral, Porites lutea. This BBD manifestation (PorBBD) presented a thick lighter-colored band, which preceded the typical BBD lesion. Whilst BBD aetiology has been intensively described worldwide, it remains unclear if corals with apparently similar lesions across coral reefs are affected by the same pathogens. Therefore, a multidisciplinary approach involving field surveys, gross lesion monitoring, histopathology and 454-pyrosequencing was employed to provide the first comprehensive characterization of this particular manifestation. Surveys conducted within two geomorphological zones over two consecutive summers and winters showed spatial and seasonal patterns consistent with those found for typical BBD. Genetic analyses suggested an uncharacteristically high level of Vibrio spp. bacterial infection within PorBBD. However, microscopic analysis revealed high densities of cyanobacteria, penetrating the compromised tissue as well as the presence of basophilic bodies resembling bacterial aggregates in the living tissue, adjacent to the bacterial mat. Additionally, classical BBD-associated cyanobacterial strains, genetically related to Pseudoscillatoria coralii and Roseofilum reptotaenium were identified and isolated and the presence of sulfate-reducers or sulfide-oxidizers such as Desulfovibrio and Arcobacter, previously shown to be associated with anoxic microenvironment within typical BBD was also observed, confirming that PorBBD is a manifestation of classical BBD.

Identification of a bacterial pathogen associated with Porites white patch syndrome in the Western Indian Ocean.

Porites white patch syndrome (PWPS) is a coral disease recently described in the Western Indian Ocean. This study aimed to isolate and identify potential pathogens associated with PWPS utilizing both culture and nonculture screening techniques and inoculation trials. A total of 14 bacterial strains (those dominant in disease lesions, absent or rare in healthy tissues and considered potential pathogens in a previous study) were cultured and used to experimentally inoculate otherwise healthy individuals in an attempt to fulfil Henle-Koch's postulates. However, only one (P180R), identified as closely related (99-100% sequence identity based on 1.4 kb 16S RNA sequence) to Vibrio tubiashii, elicited signs of disease in tank experiments. Following experimental infection (which resulted in a 90% infection rate), the pathogen was also successfully re-isolated from the diseased tissues and re-inoculated in healthy corals colonies, therefore fulfilling the final stages of Henle-Koch's postulates. Finally, we report that PWPS appears to be a temperature-dependent disease, with significantly higher tissue loss (anova: d.f. = 2, F = 39.77, P < 0.01) occurring at 30 °C [1.45 ± 0.85 cm(2) per day (mean ± SE)] compared to ambient temperatures of 28 and 26 °C (0.73 ± 0.80 cm(2) per day (mean ± SE) and 0.51 ± 0.50 cm(2) per day (mean ± SE), respectively).

Identification and prevalence of coral diseases on three Western Indian Ocean coral reefs.

Coral diseases have caused a substantial decline in the biodiversity and abundance of reef-building corals. To date, more than 30 distinct diseases of scleractinian corals have been reported, which cause progressive tissue loss and/or affect coral growth, reproductive capacity, recruitment, species diversity and the abundance of reef-associated organisms. While coral disease research has increased over the last 4 decades, very little is known about coral diseases in the Western Indian Ocean. Surveys conducted at multiple sites in Reunion, South Africa and Mayotte between August 2010 and June 2012 revealed the presence of 6 main coral diseases: black band disease (BBD), white syndrome (WS), pink line syndrome (PLS), growth anomalies (GA), skeleton eroding band (SEB) and Porites white patch syndrome (PWPS). Overall, disease prevalence was higher in Reunion (7.5 ± 2.2%; mean ± SE) compared to South Africa (3.9 ± 0.8%) and Mayotte (2.7 ± 0.3%). Across locations, Acropora and Porites were the genera most susceptible to disease. Spatial variability was detected in both Reunion and South Africa, with BBD and WS more prevalent on shallow than deep reefs. There was also evidence of seasonality in 2 diseases: the prevalence of BBD and WS was higher in summer than winter. This was the first study to investigate the ecology of coral diseases, providing both qualitative and quantitative data, on Western Indian Ocean reefs, and surveys should be expanded to confirm these patterns.

Bacterial communities associated with Porites white patch syndrome (PWPS) on three western Indian Ocean (WIO) coral reefs.

The scleractinian coral Porites lutea, an important reef-building coral on western Indian Ocean reefs (WIO), is affected by a newly-reported white syndrome (WS) the Porites white patch syndrome (PWPS). Histopathology and culture-independent molecular techniques were used to characterise the microbial communities associated with this emerging disease. Microscopy showed extensive tissue fragmentation generally associated with ovoid basophilic bodies resembling bacterial aggregates. Results of 16S rRNA sequence analysis revealed a high variability between bacterial communities associated with PWPS-infected and healthy tissues in P. lutea, a pattern previously reported in other coral diseases such as black band disease (BBD), white band disease (WBD) and white plague diseases (WPD). Furthermore, substantial variations in bacterial communities were observed at the different sampling locations, suggesting that there is no strong bacterial association in Porites lutea on WIO reefs. Several sequences affiliated with potential pathogens belonging to the Vibrionaceae and Rhodobacteraceae were identified, mainly in PWPS-infected coral tissues. Among them, only two ribotypes affiliated to Shimia marina (NR043300.1) and Vibrio hepatarius (NR025575.1) were consistently found in diseased tissues from the three geographically distant sampling localities. The role of these bacterial species in PWPS needs to be tested experimentally.