11th ICRS Abstract book - Nova Southeastern University

8.242
Microbial Community Profile Of Coral Mucus From Recovering porites Ulcerative
White Spot Disease
Miahnie Joy PUEBLOS* 1 , Wolfgang REICHARDT 1
1 The Marine Science Institute, University of the Philippines, Quezon City, Philippines
The genus Porites has been reported as a dominant host to most of the diseases found in
the Philippines, Porites Ulcerative White Spot Disease (PUWSD), of which is most
prevalent. This study aims at observing changes in the microbial community in the
mucus of both healthy and recovering colonies of Porites cylindrica using a molecular
tool like Fluorescent in situ Hybridization (FISH) and a culture-dependent method such
as plate counts. Mucus used for FISH was sampled from a coral colony via suction with
the use of a sterile syringe while mucus used for plate counts was harvested by “milking,”
which involves exposing colonies to open air to induce the production of mucus and
gentle spraying of sterile salt water. Preliminary observations under FISH, showed an
increase and decrease of Vibrio species profile during the recovery period. In the plate
count assay, mucus from healthy Porites sp. was incorporated into glycerol artificial
seawater agar, to test against potentially invasive microbes found in the ambient water
column, fish mariculture water and bacterial strains such as presumptive Vibrio isolated
from PUWS. Plate counts on mucus-incorporated saltwater-glycerol agar media versus
the presumptive Vibrio showed antibacterial properties that may possibly explain the
recovery phenomenon observed in Porites corals affected by PUWSD. Both tools used
in this study showed the possible role of Vibrio in PUWSD. Further studies should
demonstrate how extensive this role is and show whether it is the causative or the
recovery agent.
8.243
Anti-Microbial And Antibiotic Resistance Of vibrio Coralliilyticus And Sister
Phylotypes
Maria VIZCAINO* 1,2 , Gregory S. MATTHEWS 3,4 , Nikole E. KIME 1,2 , Katherine
WILLIAMS 1,2 , Peter D.R. MOELLER 1,2 , Pamela J. MORRIS 1,2
1 Marine Biomedicine and Environmental Sciences Center, Medical University of South
Carolina, Charleston, SC, 2 Hollings Marine Laboratory, Charleston, 3 Hollings Marine
Laboratory, Charleston, SC, 4 Grice Marine Laboratory, College of Charleston, Charleston
Vibrio coralliilyticus ATCC BAA-450, a temperature-dependent coral pathogen first
isolated from the Indian Ocean and Red Sea, induces bleaching in Pocillopora damicornis
at temperatures greater than 24.5°C. Currently, our research group is having the genome
of this coral pathogen sequenced and is examining it’s proteome at 24°C and 27°C. In
addition to these two approaches, we are comparing the anti-microbial and antibiotic
resistance and susceptibility profiles of this V. coralliilyticus strain and the sister
phylotypes isolated from the southern coast of Puerto Rico. First, five bacterial isolates,
obtained from the surface mucopolysaccharide layer of visually-diseased
Pseudopterogorgia americana, showed 16S rDNA homology to V. coralliilyticus.
Genomic profiling (REP-PCR) demonstrated that one isolate had a similar profile to
ATCC BAA-450, while the other four were different (but similar to one another).
Second, we examined whether bacteria isolated from healthy and diseased P. americana
colonies were able to inhibit V. coralliilyticus grown at 24°C and 27°C. We observed
that twelve (12/140) isolates inhibited V. coralliilyticus at 24°C, while only five strains
inhibited growth at 27°C. Next, four of the sister phylotypes were screened against coral
isolates that exhibited bioactivity in prior anti-microbial tests. Similarly to the ATCC
strain, four of the sister phylotypes showed a high level of resistance to the anti-microbial
activity of the coral isolates. This observed resistance increased at 27°C. Lastly, V.
coralliilyticus ATCC BAA-450 and the four sister phylotypes were screened against 26
known antibiotics; and they all exhibited similar resistance and susceptibility profiles. To
summarize, both V. coralliilyticus and Caribbean phylotypes showed similar levels of
resistance in our anti-microbial (temperature-dependent) and antibiotic assays. This
study will further contribute to our understanding of the pathogenicity of V.
coralliilyticus and the similarity with sister phylotypes in a broader ecological context.
Poster Mini-Symposium 8: Coral Microbial Interactions
324