Project by Amy Huynh, Dylan Sonett, Mauri Malta, Dr. Jesse Zaneveld. Partnered with the Global Coral Microbiome Project. Presented in 2019 at the University of Washington Bothell Biology Presentations.

1. SEASONAL EFFECTS ON
CORAL MICROBIOMES
Amy Huynh, Mauri Malta, and Dylan Sonett
Lizard Island.com.au

2. THE IMPORTANCE OF CORAL REEFS
NATURAL
BARRIER
RELATIONSHIP
WITH FISH
ECONOMICAL
DEVELOPMENT
Serves as a natural
barrier that protects
coastal communities
and shorelines from
hazards such as:
Storms
Waves
Erosion
Provide a:
Home
Nursery ground
Food source
To many coral reef fish
species
Support commercial
fisheries through tourism
and recreation
Coral reef tourism is
worth $36 billion to the
travel industry (nature.org)

3. (Adapted from Thurber et al. 2009)
RELATIONSHIP BETWEEN CORAL AND
MICROBES
Carbon source
Nutrient sequestration
Fix Nitrogen
Antibiotic production
Prevent Pathogens

4. SEASONS ALTER CORAL MICROBE
Rising seawater temperatures cause worldwide
decline of coral reef
Disease and coral bleaching
DMSP is a driver of V. coralliiyticus
Coral mucous is rich in DMSP
Vibrio coralliilyticus (coral pathogen)
uses chemotaxis to target the mucus
of coral hosts
Yellow gradient = DMSP
Pathogens use chemical gradient to
navigate towards the coral
In heat-stressed coral, DMSP concentrations
increased
Increased seawater temperature leads to an
increase in coral pathogens
(Garren et. al 2013)

5. Australian Marine Conservation Society
Coral microbiome changes from winter to summer
Coral bleaching and disease is more common in the summer
Long periods of thermal stress can kill coral by driving widespread coral bleaching
and coral disease outbreaks
Warmer temperature reduces the stability of the microbiome
Nutrient runoff can disrupt whole ecosystems
(Zaneveld et. al 2016)
SEASONS EFFECT CORALS

6. GLOBAL CORAL MICROBIOME
PROJECT
Project aims to determine the genome sequences and
metabolic capabilities of key coral bacteria
Collected data from coral in Lizard Island, Australia
Researchers collected data for Summer and Winter
months
Global Coral Microbiome Project, Pinterest

7. SEASONAL ENVIRONMENTAL CHANGE
X-axis: Months (Jul-Feb)
Y-axis: Degrees in Celsius
10°F temperature change
from winter to summer
Summer: 29°C/84°F
Winter: 24°C/74°F
Australian Institute of Marine Science

8. METHODS
GCMP DATA
GATHERING

9. METHODS
GCMP DATA
GATHERING
16S rRNA
SEQUENCING

10. 16S rRNA ANALYSIS
A component of the 30S
small subunit of a ribosome
Used in reconstructing
phylogenies because of the
slow rates of evolution in the
region of the gene
EzBioCloud

11. METHODS
GCMP DATA
GATHERING
16S rRNA
SEQUENCING
FILTER DATA TO
LIZARD ISLAND
SAMPLES
Sophie Diao, Applied and Environmental Microbiology, Vicam

12. METHODS
RAREFACTION

13. RAREIFY TO 1000 SEQUENCES PER SAMPLE

14. METHODS
RAREFACTION QIIME ALPHA +
BETA DIVERSITY

15. QIIME ANALYSIS
QIIME2 is a collection of packages used to analyze and manipulate DNA
sequence data
Computational tool for working with massive datasets
Generates diversity metrics
Runs statistical test such as Kruskal-Wallace and PERMANOVA
Attach taxonomic information to data

16. Adapted from University of Idaho
ClipartMax, DeviantArt
SITE A SITE B
ALPHA DIVERSITY
Diversity in a particular
environment or sample
BETA DIVERSITY
Difference in species across a
landscape
Site A=3 species
Site B=1 species
A VS B=2 species
3 species altogether – 1 shared

17. METHODS
RAREFACTION QIIME ALPHA +
BETA DIVERSITY
OVERALL DIVERSITY
DIVERSITY BY
COMPARTMENT
DIVERSITY BY
SPECIES
QIIME2
TAXONOMIC
ANALYSIS

18. Hypothesis
Results
Warmer
months
Increased diversity in
coral microbiome
Higher diversity in species of bacteria in the summer
Larger variation between samples in the summer
OVERALL
DIVERSITY

19. Results
Hypothesis
OVERALL
DIVERSITY

20. p = 0.001
(PERMANOVA)
Results
Hypothesis
OVERALL
DIVERSITY

21. DIVERSITY BY
COMPARTMENT
Microbiomes of each compartment demonstrate different
responses to seasonal change
Mucus would exhibit the greatest change in alpha
diversity
Hypothesis
Key Information
Results
Summary

22. Hypothesis
Results
UC San Diego
Coral compartments differ in their
microbiome
Summary
Key Information
DIVERSITY BY
COMPARTMENT

23. Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

24. Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

25. Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

26. Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

27. p = 0.001
(PERMANOVA)
Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

28. Key Information
Hypothesis
Results
TAXONOMIC
SHIFTS

29. Morton et al. 2017
Key Information
Hypothesis
Results
TAXONOMIC
SHIFTS

30. Adapted from Morton et al. 2017
Key Information
Hypothesis
Results
TAXONOMIC
SHIFTS

31. Genus Vibrio will be a greater proportion of
mucus microbes in the summer compared to
the winter.
Key Information
Results
Hypothesis
TAXONOMIC
SHIFTS

35. Genera
Caulobacter,
Sphingobium,
Blastomonas, and
Pseudomonas
increased the most
in relative
abundance from
winter to summer.
Key Information
Hypothesis
Summary
Results
DIVERSITY BY
COMPARTMENT

36. Increase in g. Sphigobium,
Pseudomonas,
Caulobacter, Blastomonas
plus f. Comamonadaceae
Decrease in unknown o.
Stramenopiles
Unknown f. Vibrionaceae
did not reach statistical
significance
ANCOM
Hypothesis
Key Information
Summary
Results
DIVERSITY BY
COMPARTMENT

37. ↑ g. Sphigobium,
Pseudomonas, Caulobacter,
Blastomonas, f.
Comamonadaceae
↓ in o. Stramenopiles
No change in f. Vibrionaceae
Relative abundance
Temperature tree a better fit
than correlation clustering or
Greengenes phylogeny
Key Information
Hypothesis
Results
Summary
DIVERSITY BY
COMPARTMENT

38. Doug Walsh / TwoFarGone.com
Key Information
Hypothesis
Results
Summary
DIVERSITY BY
SPECIES

39. The change in microbiomes is dependent on
host species and life history strategy
Stress resistant coral will exhibit less change
in alpha diversity than generalist or weedy
corals from winter to summer.
Key Information
Results
Summary
Hypothesis
DIVERSITY BY
SPECIES

40. Key Information
Hypothesis
Summary
Results
DIVERSITY BY
SPECIES

41. Key Information
Hypothesis
Summary
Results
DIVERSITY BY
SPECIES

42. Key Information
Hypothesis
Summary
Results
DIVERSITY BY
FUNCTIONAL GROUP

43. Information + Results
Experiment by
Dr. Vega Thurber
& Dr. Burkepile
Florida Keys
Mucus layer
2009 to 2012
Zaneveld et al. 2016
DIVERSITY IN
ANOTHER DATA SET

44. ● Decrease in alpha diversity from winter to summer
○ All samples
○ Mucus > Tissue
○ Turbinaria reniformis, Fungid species, Diploastrea heliopora
○ Stress-tolerant species
● Summer mucus samples are distinct from all other samples
● Change in relative abundance:
○ Increase of g. Sphigobium, Pseudomonas, Caulobacter,
Blastomonas, f. Comamonadaceae
○ Decrease of Stramenopiles
● Temperature tree a better fit to differential abundance than
correlation clustering or Greengenes phylogeny
● Opposite trend in alpha diversity in Florida Keys experiment
Key Information
Hypothesis
Results
Summary
SEASONAL
CHANGES

45. Wallpaper access