16SrDNA Pyrosequencing of the Mediterranean Gorgonian Paramuricea clavata Reveals a Link among Alterations in Bacterial Holobiont Members, Anthropogenic Influence and Disease Outbreaks

PLoS One. 2013 Jun 26;8(6):e67745. doi: 10.1371/journal.pone.0067745. Print 2013.

Abstract

Mass mortality events of benthic invertebrates in the Mediterranean Sea are becoming an increasing concern with catastrophic effects on the coastal marine environment. Sea surface temperature anomalies leading to physiological stress, starvation and microbial infections were identified as major factors triggering animal mortality. However the highest occurrence of mortality episodes in particular geographic areas and occasionally in low temperature deep environments suggest that other factors play a role as well. We conducted a comparative analysis of bacterial communities associated with the purple gorgonian Paramuricea clavata, one of the most affected species, collected at different geographic locations and depth, showing contrasting levels of anthropogenic disturbance and health status. Using massive parallel 16SrDNA gene pyrosequencing we showed that the bacterial community associated with healthy P. clavata in pristine locations was dominated by a single genus Endozoicomonas within the order Oceanospirillales which represented ∼90% of the overall bacterial community. P. clavata samples collected in human impacted areas and during disease events had higher bacterial diversity and abundance of disease-related bacteria, such as vibrios, than samples collected in pristine locations whilst showed a reduced dominance of Endozoicomonas spp. In contrast, bacterial symbionts exhibited remarkable stability in P. clavata collected both at euphotic and mesophotic depths in pristine locations suggesting that fluctuations in environmental parameters such as temperature have limited effect in structuring the bacterial holobiont. Interestingly the coral pathogen Vibrio coralliilyticus was not found on diseased corals collected during a deep mortality episode suggesting that neither temperature anomalies nor recognized microbial pathogens are solely sufficient to explain for the events. Overall our data suggest that anthropogenic influence may play a significant role in determining the coral health status by affecting the composition of the associated microbial community. Environmental stressful events and microbial infections may thus be superimposed to compromise immunity and trigger mortality outbreaks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anthozoa / microbiology*
  • Bacteria / classification
  • Bacteria / genetics*
  • Bacteria / isolation & purification
  • Bacterial Infections / epidemiology*
  • Bacterial Infections / genetics
  • Bacterial Infections / microbiology
  • DNA, Bacterial / genetics*
  • DNA, Ribosomal / genetics*
  • Disease Outbreaks*
  • Mediterranean Sea
  • Sequence Analysis, DNA
  • Symbiosis

Substances

  • DNA, Bacterial
  • DNA, Ribosomal

Grants and funding

This work was promoted and funded by the Committee for Research and Exploration of the National Geographic Society (grant Grant Agreement 8876-11), by the Italy-Israel joint research Programme, Directorate General Political and Affairs of the Foreign Affairs, Italy (contract number 3697760016) and by PRIN 2008 Italian funds (2008YBE-321 ANX_002). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.