Determination of <i>Philaster</i> sp presence.

<p>PCR product check using <i>Philaster</i> sp specific primers (BrB-F-171 and BrB-R-1721) on healthy (H), sloughed tissue (ST), untreated (WSU) and treated WS (WST) samples, carried out using gel electrophoresis on a 1% agarose gel. P indicates bands representing <i>Philaster</i> sp. presence.</p

Bacterial abundance of <i>Echinopora lamellosa</i> samples.

<p>Mean bacterial abundance for apparently healthy (H), untreated White Syndrome-affected (WSU) and treated White Syndrome-affected (WST) <i>Echinopora lamellosa</i> samples. Error bars represent SE.</p

Neighbour-joining consensus tree of partial 18S rRNA gene sequences of 4 ciliate species found in <i>Echinopora lamellosa</i> samples affected by White Syndrome in this aquarium experiment.

<p>Sequence alignment was carried out using MUSCLE and the neighbour-joining tree was constructed in TreeDyn (via phylogeny.fr), using the Tamura genetic distance model [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121780#pone.0121780.ref040" target="_blank">40</a>] with an opalinid protist; <i>Opalina ranarum</i> (AF141970) as the outgroup, as used by Sweet and Bythell [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121780#pone.0121780.ref018" target="_blank">18</a>]</p

Ciliate 18S rRNA analysis of <i>Echinopora lamellosa</i> samples.

<p>Ciliate 18S rRNA gene fingerprints (represented on Denaturing Gradient Gel Electrophoresis) of healthy (H), sloughed tissue (ST), untreated (WSU) and treated WS (WST) samples. M = marker lane.</p

Heat-map representing the relative abundance of identified (GenBank ID) bacteria and archaea taxa associated with H, ST, UWS and TWS samples.

<p>Identification and sequence similarity was based on 161 bp sequences obtained from excised denaturing gradient gel electrophoresis (DGGE) bands. Relative abundance values are of scaled arbitrary units, based on relative density of DGGE bands in BioNumerics (3.5) analysis. Absence of a particular band is represented by a zero.</p

Antimicrobial activity of apparently healthy and White Syndrome-affected <i>Echinopora lamellosa</i> samples.

<p>Antimicrobial activity of apparently healthy and White Syndrome-affected <i>Echinopora lamellosa</i> samples.</p

Microbial analysis of <i>Plectropomus leopardus</i> samples; a) Bacterial 16S rRNA gene fingerprints (represented on Denaturing Gradient Gel Electrophoresis) of fish mucus (SWB) and tissue samples (TSU), standardised for gel-to-gel comparison using BioNumerics; b) resin embed histological section of a healthy fish, stained with the general DNA stain 4′6-diamidino-2-phenylindole (DAPI); c) histological section of the lesion on a diseased fish stained with DAPI, both showing no bacteria within the dermis suggesting the bacteria present in (a) are localised within the mucus layer not within the tissues.

<p>Scale bars = 10 µm.</p

Microscopic images of <i>Plectropomus leopardus</i> tissues; a) Scanning Electron Micrograph (SEM) of the healthy tissue; b) SEM of the lesion.

<p>MGC = mucus goblet cells, M = mucus. c) Light microscope image of a healthy scale and d) light microscope image of a diseased scale, showing disorganisation of natural melanin patterns seen in (c). Scales bars = 10 µm.</p

Lesions were present on approximately 15% of the sampled population of <i>Plectropomus leopardus</i>; a) affected individual showing <10% coverage of body surface; b) <i>P. leopardus</i> with almost complete coverage >90%; c) healthy tissue under light microscope and d) the lesion.

<p>Scale bars = 20 µm.</p

Transmission Electron Micrographs of different samples of <i>P. leopardus</i> exhibiting; (a) healthy tissue showing the two cell types (A & B) associated in the dermis along the collagenous basal membrane (CBM).

<p>Cell A shows localisation of melanosomes and Cell B shows absence of melanosomes in the same area. (b) Lesion showing disorganisation of pleomorphic cells (A & B) with an increase in number and spread of melanosomes. This lesion is an example of a <i>P. leopardus</i> suffering from stage II melanoma, where the melanosomes are restricted to the dermis. Scale bars = 10 µm.</p