Chemical Mechanisms of Disease Resistance

in Hawai’ian Corals

Deborah Gochfeld1, Greta Aeby2, Jesse Miller1

1National Institute of Undersea Science and Technology,University of Mississippi

2Hawaii Institute of Marine Biology

Occurrence of infectious disease

Host

Pathogen

Environment

Occurrence of infectious disease

Host

Pathogen

Environment

0

0.5

1

1.5

2

2.5

3

3.5

4

Montipora Porites Pocillopora

overall disease prevalence (%)

Mauin=9 sites

Antimicrobial defense

Occurrence of infectious disease

Host

Pathogen

Environment

00.5

11.5

22.5

33.5

44.5

OlowaluHonolua N

KahekiliKanahena

MolokiniPuamanaMala Wharf

Ma'alaeaSugar Beach

est. prevalence (%)

Antimicrobial defense

Objectives

1) Assess differences in antimicrobial activity among three dominant Hawai’ian coral genera.

2) Assess population-level variability in antimicrobial activity of common coral species from three different sites.

3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

Montipora Porites Pocillopora

General Methods

1) Collect replicate pieces of three common coral genera (Montipora, Pocillopora, Porites) from three sites on Oahu

2) Extract corals is aqueous solvents

3) Test extracts in antimicrobial assays to determine genus- and population-level variability in antimicrobial activity

BacteriaKnown coral

pathogen

Potential marine

pathogen from human waste

Isolated from surfaces of Hawaiian

corals

Aurantimonas coralicida X

Vibrio corallyticus X

Vibrio shiloi X X

Serratia marcescens X X

Clostridium perfringens X

Yersinia enterocolitica X X

Pseudomonas nautica X

Vibrio agarivorans X

Klebsiella pneumoniae X

Species of bacteria used to test coral extracts

Progress Since May

• Collected replicate corals from three sites around Oahu: Waianae, Maile Beach, Kaneohe Bay

• Extracted corals for assays•Porites lobata (n=5 from each of 3 sites)•Pocillopora meandrina (n=5 from each of 3 sites)•Montipora capitata (n=5 from each of 3 sites)

• Collected and extracted healthy and diseased tissues from diseased Montipora capitata from Kaneohe Bay, along with healthy neighbor controls (n=5 of each)

• Performed growth inhibition assay of all aqueous extracts from Porites lobata and Pocillopora meandrina against 9 bacterial strains.

• Performed growth inhibition assay on extracts from healthy, diseased and control Montipora capitata from Kaneohe Bay to determine whether further chemical fingerprinting is warranted

Objectives

1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera.

2) Assess population-level variability in antimicrobial activity of common coral species from different sites.

3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

Anti-microbial activity of Hawaiian reef corals

0

20

40

60

80

100

A. coralicidaV. corallyticus

V. shiloi

S. marcescensY. enterocolitica

C. perfringensP. nautica

V. agarivoransK. pneumoniaebacterial species

proportion of colonies w/ antimicrobial activity(%)

P. lobataN=15

Anti-microbial activity of Hawaiian reef corals

0

20

40

60

80

100

A. coralicidaV. corallyticus

V. shiloi

S. marcescensY. enterocolitica

C. perfringensP. nautica

V. agarivoransK. pneumoniae

bacterial species

proportion of colonies w/ antimicrobial

activity (%)

P. meandrina N=15

Intraspecific variability in antimicrobial activity

Bacteria P. lobata(n=15)

Aurantimonas coralicida -12Vibrio corallyticus -10/+3Vibrio shiloi -5/+3Serratia marcescens -15Yersinia enterocolitica -10Clostridium perfringens -14Pseudomonas nautica +13Vibrio agarivorans -15Klebsiella pneumoniae -1/+3

Intraspecific variability in antimicrobial activity

Bacteria P. lobata P. meandrina(n=15) (n=15)

Aurantimonas coralicida -12 -9/+2Vibrio corallyticus -10/+3 -6/+1Vibrio shiloi -5/+3 -1/+8Serratia marcescens -15 -11/+1Yersinia enterocolitica -10 -13/+1Clostridium perfringens -14 -14/+1Pseudomonas nautica +13 -1/+11Vibrio agarivorans -15 -11/+2Klebsiella pneumoniae -1/+3 -3/+11

Intraspecific variability in disease resistance in corals

Objectives

1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera.

2) Assess population-level variability in antimicrobial activity of common coral species from different sites.

3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

Site differences in antimicrobial activity of corals

Porites lobata

Bacteria Waianae Maile Beach Kaneohe Bay

Aurantimonas coralicida -3 -4 -5Vibrio corallyticus +3 -5 -5Vibrio shiloi -3/+1 -1/+1 -1/+1Serratia marcescens -5 -5 -5Yersinia enterocolitica -5 -5 0Clostridium perfringens -4 -5 -5Pseudomonas nautica +5 +3 +5Vibrio agarivorans -5 -5 -5Klebsiella pneumoniae +2 -1 +1

antimicrobial activity (%) 55.6 68.9 57.8

Site differences in antimicrobial activity of corals

Pocillopora meandrina

Bacteria Waianae Maile Beach Kaneohe Bay

Aurantimonas coralicida -2/+1 -4 -3/+1Vibrio corallyticus -5 +4 -1Vibrio shiloi -1/+3 +5 0Serratia marcescens -3 -5 -3/+1Yersinia enterocolitica -5 -5 -3/+1Clostridium perfringens -5 -5 -4/+1Pseudomonas nautica -1/+2 +5 +4Vibrio agarivorans -4 -2/+2 -5Klebsiella pneumoniae -2/+3 -1/+3 +5

antimicrobial activity (%) 62.2 48.9 42.2

Site differences in antimicrobial activity of corals Pocillopora meandrina

Bacteria Waianae Maile Beach Kaneohe Bay

Aurantimonas coralicida -2/+1 -4 -3/+1Vibrio corallyticus -5 +4 -1Vibrio shiloi -1/+3 +5 0Serratia marcescens -3 -5 -3/+1Yersinia enterocolitica -5 -5 -3/+1Clostridium perfringens -5 -5 -4/+1Pseudomonas nautica -1/+2 +5 +4Vibrio agarivorans -4 -2/+2 -5Klebsiella pneumoniae -2/+3 -1/+3 +5

00.5

11.5

22.5

33.5

44.5

OlowaluHonolua N

KahekiliKanahena

MolokiniPuamanaMala Wharf

Ma'alaeaSugar Beach

est. prevalence (%)

land based pollution stress defensenovel pathogens

water flow inoculumsrepetitive exposure increased water temperature

Objectives

1) Assess differences in antimicrobial activity among dominant Hawai’ian coral genera.

2) Assess population-level variability in antimicrobial activity of common coral species from different sites.

3) Assess differences in antimicrobial activity and chemical constituents in healthy and diseased colonies of the same species.

Montipora white syndrome

Montipora white syndrome

Comparison of antimicrobial activity in healthy vs. diseased colonies

Healthy neighbor

Higher levels of antimicrobials?Induced defenses?

Effect of extracts from healthy & diseasedMontipora capitata on growth of bacteria

M. capitata

Bacteria control normal affected

Aurantimonas coralicida -4 -3 -4Vibrio corallyticus

Vibrio shiloi -5 -4 -4Serratia marcescens

Yersinia enterocolitica

Clostridium perfringens -5 -5 -3Pseudomonas nautica -5 -3 -5Vibrio agarivorans -5 -3 -3Klebsiella pneumoniae

antimicrobial activity (%) 96 72 76

Conclusions

Host

Pathogen

Environment

Broad spectrum antimicrobial activity

Intraspecific variability Differences within coral species among sites

Differences between healthy and diseased colony

Acknowledgements

Collections(Hawaii Institute of Marine Biology)

Megan Colwin Lian ZigtermanCasper Lazet

Sample Processing and Assays(University of Mississippi)

Shellie BaileyAmy KatzenmeyerLauren Wheeler