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Rickettsiales-like organisms (RLOs) are thought to be related to bacteria in the order Rickettsiales. They have been reported to occur in the staghorn coral (Acropora cervicornis), and this study investigated trends of infection over time, and in relation to the health of infected corals. This study focuses on tissue samples taken mostly from visibly healthy A. cervicornis thickets in Broward County, Florida, and processed for histological examination. Samples were originally collected and analyzed to document reproduction during years 2001 through 2012, and tissue loss diseases (white-band disease [WBD] types I and II, and rapid tissue loss). The presence of suspect RLOs, the presence of ovoid bacterial aggregates in the basal body wall, and the condition of the coral tissue were examined in hematoxylin and eosin (H&E) and Giemsa-stained sections. Determination was made as to whether suspect RLO infection severity, location, or the presence of bacterial aggregates are correlated with changes in tissue condition associated with WBD. To better understand progression, trends, and periodicity in bacterial presence and coral tissue health, these data were then further analyzed for potential correlation with the month, year, and average monthly nighttime sea surface temperatures (categorized into ranges above, within, or below 24–29°C) when samples were collected. The severity of suspect RLO infections and the presence of bacterial aggregates in A. cervicornis varied over time, with no correlation with the location of suspect RLOs within the polyp. High suspect RLO prevalence was correlated with normal tissue conditions, while low suspect RLO prevalence was correlated with abnormal tissue conditions. However, high prevalence of bacterial aggregates was correlated to abnormal tissue conditions. Epidermal RLO and overall suspect RLO prevalence severity scores were significantly higher among samples collected when monthly average nighttime sea surface temperatures were below 24°C in contrast to samples collected when temperatures were between 24–29°C, suggesting direct or indirect effects of sea surface temperatures on infection severity. The areas of suspect RLO intracellular bodies within infected mucocytes were measured using digital image analysis software and found to be positively correlated with worsening coral tissue condition. Semiquantitative variable scores for histoslides stained with H&E were significantly different from those stained with Giemsa, indicating that these stains cannot be used interchangeably to study the presence of bacteria and the condition of coral tissue. Overall, the results of this study indicate that infection severity of suspect RLOs and the presence of bacterial aggregates are variable and correlated with the incidence of WBD-I in A. cervicornis. However, the exact nature of this relationship remains unclear. Further studies are necessary to interpret trends detected during this analysis to develop a better understanding of what contributes to the severe tissue-loss outbreaks and mortalities of A. cervicornis.

Caribbean reef ecosystems have undergone major ecological changes in the last 30 – 40 years, with the result that ecological systems once dominated by structurally complex Acropora cervicornis and Montastraea annularis corals now consist mainly of flattened carbonate substrates with macroalgal overgrowth. A need for greater understanding of coral reef ecosystems is imperative if we are to attempt to conserve them. The threespot damselfish, Stegastes planifrons, is herbivorous damselfish species ubiquitous to Caribbean reefs, where it has been termed a keystone species. Aggressive in nature, S. planifrons defends territories of around 70 cm in diameter from other roving herbivorous fish and urchins, in apparent effort to maintain the algal resources therein for its own use. The predilection of Stegastes planifrons for basing its territories on the now Critically Endangered staghorn coral, Acropora cervicornis, and the Endangered boulder coral Montastraea annularis is well known, however the likely ecological implications of this fact have not been investigated. Using a combination of experimental and observational methodologies we examine the ecological implications of coral microhabitat choice and use by S. planifrons. We also assess the magnitude of the direct and indirect effects of S. planifrons’ territorial behaviour on macroalgal dynamics both within and outside of territory confines, at the reef-wide level. We find that coral microhabitat is a more important determinant of algal community structure than damselfish presence, and that this can be explained by a previously unrecognised effect of coral microhabitat on the grazing behaviour of roving herbivorous fishes - on which S. planifrons’ territorial behaviour has little effect. In a modification of the space availability hypothesis of Williams et al (2001) we suggest that Acropora cervicornis acts as a grazing fish „exclusion zone‟, and we further hypothesise that the existence of large stands of this coral prior to the Caribbean „phase shift‟ may have acted to concentrate the grazing pressure of excluded roving fish onto the remaining areas of the reef. We further hypothesise that the loss of such „exclusion zones‟ and accompanied effective dilution of grazing pressure may have been on a scale large enough to have been a significant underlying factor in the proliferation of macroalgae seen on modern day Caribbean reefs. In the absence of demonstrable direct or indirect effects on benthic algal communities we question the continued keystone status of S. planifrons, particularly since the status 6 was originally based on interference behaviour involving the important grazing urchin Diadema antillarum, which is now functionally absent from Caribbean reefs. Implications of the context-dependant nature of keystone status are also discussed. We find that the effect of S. planifrons on coral community may be more important than its effects on benthic algal community. In examining the factors involved in habitat coral choice we establish a significant preference for 100% live coral substrate over substrates with a supply of algal food. Territory selection was followed by a high rate of coral biting – a behaviour which has previously been shown to result in coral tissue death and the fast establishment of algal turf communities on which S. planifrons likes to feed (Kaufman 1977). We also demonstrate a novel and significant association between S. planifrons presence and disease incidence its primary habitat coral, the Critically Endangered staghorn coral Acropora cervicornis, and a significant correlation between areas of fish biting and the later onset of disease. Changes to the overall role of damselfish on today's Caribbean reefs are discussed in light of these insights.

White Band disease has devastated the staghorn coral Acropora cervicornis in recent decades, and it continues to impinge upon restoration efforts. The etiological agent(s) remain unknown as Koch’s postulates have yet to be satisfied, but disease may originate when opportunistic pathogens in the surface mucus layer exploit a stressed host. Using 16s rRNA sequencing, differences in the taxonomic diversity and relative abundances of bacteria within the mucus of A. cervicornis were documented between colonies of the same genotype, genotypes (n=8) categorized as having either high or low WBD susceptibility, and during a transplantation event. A. cervicornis colonies suspended from midwater PVC trees via monofilament were sampled for mucus, after which half of the sampled colonies were relocated to the unconsolidated sediments below. Temporal changes in the microbiome of the pelagic and benthic corals were then monitored by sampling the same apical tip over time. Incidentally, all benthic colonies for this experiment became afflicted with WBD; thereby differences in healthy vs. diseased colonies and the effects of disease progression on the microbiome were documented. Water was sampled concurrently with all mucus experiments to resolve the degree of commonality in bacterial species between the two environments, and sediments were sampled in the transplant experiment to determine if sediments may act as a pathogen reservoir. In addition, sediment samples were collected to assess site and temporal differences in the benthic microbiome along a nearshore to offshore transect off Key Largo, Florida. Irrespective of the inclusion of water operational taxonomic units (OTUs), no differences between colonies of the same genotype were observed with regards to the bacterial communities sampled from mucus in either alpha diversity metrics [species richness, Shannon, Inverse Simpson] or phylogenetic relatedness as determined by weighted unique fraction (UniFrac) were detected between colonies. However, differences were observed in the Bray-Curtis dissimiliarity matrices based on relative abundance and presence/absence of either [with and without water OTU] scenarios. Bacterial communities associated with different coral genotypes differed in species richness and Inverse Simpson in both water scenarios, as did weighted UniFrac and Bray-Curtis relative abundance and presence/absence transformed dissimilarity matrices. Alpha diversity of mucus bacteria was similar between corals of different disease-susceptibilities when water OTUs were either included or excluded, except for the Inverse Simpson index upon removal of water OTUs. Removal of aqueous bacteria also revealed significant differences between disease-susceptibility groups in Bray-Curtis relative abundance and presence/absence dissimiliarity values that was not detected with the incorporation of water OTUs. Regardless of the presence of water OTUs, weighted UniFrac was similar between corals of different disease susceptibilities. Most notably, dispersion increased in the microbiome of coral genotypes with high disease susceptibility in all cases except for the relative abundance transformed Bray-Curtis dissimilarity matrix when water OTUs were incorporated. This finding is in accordance with the Anna Karenina Principle, which states that loss of microbial regulation leads to an unpredictable microbiome in diseased individuals. In the sediment experiment, location was the only factor influencing microbiome composition. These findings may be due to the short duration of the experiment and differences between the carbonate content of the sediments and hydrological regimes between sites.

The high latitude thickets of Acropora cervicornis off Broward County flourish despite the presence of natural and anthropogenic impacts. These populations provided a unique study area in contrast to disease-stricken populations of the Florida Keys. This study used time-sequenced photographs to examine how A. cervicornis was affected by tissue loss attributed to white-band disease during 2007–2008. Variables monitored included healthy colony skeletal extension rates, diseased colony skeletal extension rates, and tissue loss. The transmissibility of the three white-band syndromes found in the Scooter and Oakland thickets was examined through tissue grafting experiments. Skeletal extension rates of healthy and diseased colonies were generally not significantly different. Mean skeletal extension for A. cervicornis colonies in Broward County was observed to be 9.6 cm/y (SD=3.95, Range: 1.02–19.9). Mean linear tissue loss from disease signs was 2.6 mm/d (SD=4.3, Range: 0.023–16.8). Although the majority of active disease lesions caused severe tissue loss upon contact with healthy branches, in 25% of the cases there was no tissue loss. Disease signs were also observed in 10% of the control grafting trials. A. cervicornis thickets in Broward County were growing at rates similar to those observed in this species elsewhere in Florida, but faster than other areas of the Western Atlantic. Tissue loss rate from disease lesions was lower than reported elsewhere. White-band disease and/or other tissue loss syndromes are always present in Broward County, but the low prevalence of affected colonies, inconsistent transmission of a presumptive agent that causes the disease signs, and optimum branch skeletal extension seems to limit effects on the thickets. Results of this research are significant as the current protected status of acroporid corals no longer allows manipulative research such as coral grafting for transmissibility of potential disease pathogens.