First Record of Black Band Disease in the Hawaiian Archipelago: Response, Outbreak Status, Virulence, and a Method of Treatment

A high number of coral colonies, Montipora spp., with progressive tissue loss were reported from the north shore of Kaua‘i by a member of the Eyes of the Reef volunteer reporting network. The disease has a distinct lesion (semi-circular pattern of tissue loss with an adjacent dark band) that was first observed in Hanalei Bay, Kaua‘i in 2004. The disease, initially termed Montipora banded tissue loss, appeared grossly similar to black band disease (BBD), which affects corals worldwide. Following the initial report, a rapid response was initiated as outlined in Hawai‘i’s rapid response contingency plan to determine outbreak status and investigate the disease. Our study identified the three dominant bacterial constituents indicative of BBD (filamentous cyanobacteria, sulfate-reducing bacteria, sulfide-oxidizing bacteria) in coral disease lesions from Kaua‘i, which provided the first evidence of BBD in the Hawaiian archipelago. A rapid survey at the alleged outbreak site found disease to affect 6-7% of the montiporids, which is higher than a prior prevalence of less than 1% measured on Kaua‘i in 2004, indicative of an epizootic. Tagged colonies with BBD had an average rate of tissue loss of 5.7 cm2/day over a two-month period. Treatment of diseased colonies with a double band of marine epoxy, mixed with chlorine powder, effectively reduced colony mortality. Within two months, treated colonies lost an average of 30% less tissue compared to untreated controls

Vibrio coralliilyticus Strain OCN008 Is an Etiological Agent of Acute Montipora White Syndrome

Identification of a pathogen is a critical first step in the epidemiology and subsequent management of a disease. A limited number of pathogens have been identified for diseases contributing to the global decline of coral populations. Here we describe Vibrio coralliilyticus strain OCN008, which induces acute Montipora white syndrome (aMWS), a tissue loss disease responsible for substantial mortality of the coral Montipora capitata in Ka ne‘ohe Bay, Hawai‘i. OCN008 was grown in pure culture, recreated signs of disease in experimentally infected corals, and could be recovered after infection. In addition, strains similar to OCN008 were isolated from diseased coral from the field but not from healthy M. capitata. OCN008 repeatedly induced the loss of healthy M. capitata tissue from fragments under laboratory conditions with a minimum infectious dose of between 107 and 108 CFU/ml of water. In contrast, Porites compressa was not infected by OCN008, indicating the host specificity of the pathogen. A decrease in water temperature from 27 to 23°C affected the time to disease onset, but the risk of infection was not significantly reduced. Temperature-dependent bleaching, which has been observed with the V. coralliilyticus type strain BAA-450, was not observed during infection with OCN008. A comparison of the OCN008 genome to the genomes of pathogenic V. coralliilyticus strains BAA-450 and P1 revealed similar virulence-associated genes and quorum-sensing systems. Despite this genetic similarity, infections of M. capitata by OCN008 do not follow the paradigm for V. coralliilyticus infections established by the type strain

Ecological and molecular characterization of a coral black band disease outbreak in the Red Sea during a bleaching event

Black Band Disease (BBD) is a widely distributed and destructive coral disease that has been studied on a global scale, but baseline data on coral diseases is missing from many areas of the Arabian Seas. Here we report on the broad distribution and prevalence of BBD in the Red Sea in addition to documenting a bleaching-associated outbreak of BBD with subsequent microbial community characterization of BBD microbial mats at this reef site in the southern central Red Sea. Coral colonies with BBD were found at roughly a third of our 22 survey sites with an overall prevalence of 0.04%. Nine coral genera were infected including Astreopora, Coelastrea, Dipsastraea, Gardineroseris, Goniopora, Montipora, Pavona, Platygyra, and Psammocora. For a southern central Red Sea outbreak site, overall prevalence was 40 times higher than baseline (1.7%). Differential susceptibility to BBD was apparent among coral genera with Dipsastraea (prevalence 6.1%), having more diseased colonies than was expected based on its abundance within transects. Analysis of the microbial community associated with the BBD mat showed that it is dominated by a consortium of cyanobacteria and heterotrophic bacteria. We detected the three main indicators for BBD (filamentous cyanobacteria, sulfate-reducing bacteria (SRB), and sulfide-oxidizing bacteria (SOB)), with high similarity to BBD-associated microbes found worldwide. More specifically, the microbial consortium of BBD-diseased coral colonies in the Red Sea consisted of Oscillatoria sp. (cyanobacteria), Desulfovibrio sp. (SRB), and Arcobacter sp. (SOB). Given the similarity of associated bacteria worldwide, our data suggest that BBD represents a global coral disease with predictable etiology. Furthermore, we provide a baseline assessment of BBD disease prevalence in the Red Sea, a still understudied region

Differential Susceptibility Manuscript_RawData_FigShare.docx.xlsx

Raw data for publication entitled:<div>Trade-offs in disease and bleaching susceptibility among two color morphs of the Hawaiian reef coral, <i>Montipora capitata</i><br></div><div><i><br></i></div><div>Authors:</div><div>Shore-Maggio, A; Callahan, S; Aeby, G </div

Phylogenetic relationship of <i>Beggiatoa</i> spp. isolated from disease lesions from Kaua‘i to other <i>Beggiatoa</i>.

<p>A phylogenetic tree was generated using the Maximum Likelihood method with sequences from 14 other <i>Beggiatoa</i> and closely related sulfur-oxidizing strains including representative type strains from the Thiotrichaceae family. No 16S rRNA gene sequences from <i>Beggiatoa</i> found in BBD from other regions were available for comparison. The tree with the highest log likelihood is shown, and 1000 bootstrap replicates were used. NCBI accession numbers are in brackets, and bootstrap values are indicated at branch nodes. Scale bar represents five substitutions per 100 nucleotide positions.</p

The lesion occlusion method of disease treatment.

<p>A) Infected <i>M</i>. <i>capitata</i> before treatment. B) Colony with marine epoxy over lesion and with a band of epoxy placed as a “firebreak” approximately 5 cm away. C) Same colony two months post-treatment. Note that the marine epoxy is overgrown with algae and the edge of the lesion is starting to grow over the epoxy. D) Control infected <i>M</i>. <i>capitata</i> before marking with marine epoxy. E) Colony with marine epoxy approximately 5 cm behind lesion. F) Control colony after two months.</p

Phylogenetic relationship of cyanobacterium OCN074, isolated from disease lesions from Kaua‘i, to other cyanobacteria.

<p>All 16S rRNA gene sequences from the three Kaua‘i BBD cyanobacterial isolates were identical. A phylogenetic tree was generated using the Maximum Likelihood method. The ‘*’ indicates cyanobacteria associated with BBD of Palau (*[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref042" target="_blank">42</a>]) the Red Sea (**[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref041" target="_blank">41</a>]), and the Florida Keys (***[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref043" target="_blank">43</a>]). The tree with the highest log likelihood is shown, and 1000 bootstrap replicates were used. NCBI accession numbers are in brackets and bootstrap values are indicated at branch nodes. Scale bar represents two substitutions per 100 nucleotide positions.</p

Phylogenetic relationship of sulfate-reducing bacteria from disease lesions from Kaua‘i to other sulfate-reducing bacteria.

<p>A phylogenetic tree was generated using the Maximum Likelihood method with a reference sequence library built from 25 known <i>dsrA</i> gene sequences used as a proxy for the presence of sulfate-reducing bacteria [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref023" target="_blank">23</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref033" target="_blank">33</a>]. The ‘*’ indicates gene sequences from BBD samples previously published from the Great Barrier Reef (* [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref033" target="_blank">33</a>]), and the Red Sea (** [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120853#pone.0120853.ref023" target="_blank">23</a>]). The tree with the highest log likelihood is shown, and 1000 bootstrap replicates were used. NCBI accession numbers are in brackets, and bootstrap values are indicated at branch nodes. Scale bar represents two substitutions per 100 nucleotide positions.</p

<i>Montipora capitata</i> colony with signs of black band disease.

<p><i>Montipora capitata</i> colony with signs of black band disease.</p

Brightfield image of Beggiatoa sp. filaments removed from disease lesions.

<p>Image taken at 60x magnification. Black bar represents 20 μm.</p