Antibiotics reduce Pocillopora coral‐associated bacteria diversity, decrease holobiont oxygen consumption and activate immune gene expression

Corals are important models for understanding invertebrate host-microbe interactions; however, to fully discern mechanisms involved in these relationships, experimental approaches for manipulating coral-bacteria associations are needed. Coral-associated bacteria affect holobiont health via nutrient cycling, metabolic exchanges and pathogen exclusion, yet it is not fully understood how bacterial community shifts affect holobiont health and physiology. In this study, a combination of antibiotics (ampicillin, streptomycin and ciprofloxacin) was used to disrupt the bacterial communities of 14 colonies of the reef framework-building corals Pocillopora meandrina and P. verrucosa, originally collected from Panama and hosting diverse algal symbionts (family Symbiodiniaceae). Symbiodiniaceae photochemical efficiencies and holobiont oxygen consumption (as proxies for coral health) were measured throughout a 5-day exposure. Antibiotics altered bacterial community composition and reduced alpha and beta diversity, however, several bacteria persisted, leading to the hypothesis that these bacteria are either antibiotics resistant or occupy internal niches that are shielded from antibiotics. While antibiotics did not affect Symbiodiniaceae photochemical efficiency, antibiotics-treated corals had lower oxygen consumption rates. RNAseq revealed that antibiotics increased expression of Pocillopora immunity and stress response genes at the expense of cellular maintenance and metabolism functions. Together, these results reveal that antibiotic disruption of corals' native bacteria negatively impacts holobiont health by decreasing oxygen consumption and activating host immunity without directly impairing Symbiodiniaceae photosynthesis, underscoring the critical role of coral-associated bacteria in holobiont health. They also provide a baseline for future experiments that manipulate Pocillopora corals' symbioses by first reducing the diversity and complexity of coral-associated bacteria.

[1]  Shenmin Zhang,et al.  Cultured Bacteria Provide Insight into the Functional Potential of the Coral-Associated Microbiome , 2022, mSystems.

[2]  N. Traylor-Knowles,et al.  Antibiotics Alter Pocillopora Coral-Symbiodiniaceae-Bacteria Interactions and Cause Microbial Dysbiosis During Heat Stress , 2022, Frontiers in Marine Science.

[3]  F. Houlbrèque,et al.  Highly Variable and Non-complex Diazotroph Communities in Corals From Ambient and High CO2 Environments , 2021, Frontiers in Marine Science.

[4]  S. Bonnet,et al.  Microbes support enhanced nitrogen requirements of coral holobionts in a high CO2 environment , 2021, Molecular ecology.

[5]  K. Nelson,et al.  Coral microbiome manipulation elicits metabolic and genetic restructuring to mitigate heat stress and evade mortality , 2021, Science Advances.

[6]  J. Robbins,et al.  Characterizing the culturable surface microbiomes of diverse marine animals , 2021, FEMS microbiology ecology.

[7]  R. Peixoto,et al.  Delivering Beneficial Microorganisms for Corals: Rotifers as Carriers of Probiotic Bacteria , 2020, Frontiers in Microbiology.

[8]  M. Kawachi,et al.  A New Dinoflagellate Genome Illuminates a Conserved Gene Cluster Involved in Sunscreen Biosynthesis , 2020, Genome biology and evolution.

[9]  Åsa K. Björklund,et al.  Prototypical pacemaker neurons interact with the resident microbiota , 2020, Proceedings of the National Academy of Sciences.

[10]  D. Crawford,et al.  An Innovative Setup for High-Throughput Respirometry of Small Aquatic Animals , 2020, bioRxiv.

[11]  Kohske Takahashi,et al.  Welcome to the Tidyverse , 2019, J. Open Source Softw..

[12]  R. Irizarry ggplot2 , 2019, Introduction to Data Science.

[13]  K. Guillemin,et al.  Evolutionary "Experiments" in Symbiosis: The Study of Model Animals Provides Insights into the Mechanisms Underlying the Diversity of Host-Microbe Interactions. , 2019, BioEssays : news and reviews in molecular, cellular and developmental biology.

[14]  M. V. van Oppen,et al.  Experimental Inoculation of Coral Recruits With Marine Bacteria Indicates Scope for Microbiome Manipulation in Acropora tenuis and Platygyra daedalea , 2019, Front. Microbiol..

[15]  Senjie Lin,et al.  Altered Immune Landscape and Disrupted Coral-Symbiodinium Symbiosis in the Scleractinian Coral Pocillopora damicornis by Vibrio coralliilyticus Challenge , 2019, Front. Physiol..

[16]  M. Matz,et al.  Contrasting effects of Symbiodinium identity on coral host transcriptional profiles across latitudes , 2018, Molecular ecology.

[17]  P. Bongaerts,et al.  Adaptation to reef habitats through selection on the coral animal and its associated microbiome , 2018, Molecular ecology.

[18]  Yu Song,et al.  Anti-quorum Sensing Activities of Selected Coral Symbiotic Bacterial Extracts From the South China Sea , 2018, Front. Cell. Infect. Microbiol..

[19]  S. Fraune,et al.  Predicted Bacterial Interactions Affect in Vivo Microbial Colonization Dynamics in Nematostella , 2018, Front. Microbiol..

[20]  C. Wild,et al.  Dominance of Endozoicomonas bacteria throughout coral bleaching and mortality suggests structural inflexibility of the Pocillopora verrucosa microbiome , 2018, Ecology and evolution.

[21]  Jean M. Macklaim,et al.  Microbiome Datasets Are Compositional: And This Is Not Optional , 2017, Front. Microbiol..

[22]  Yinglin Xia,et al.  Hypothesis testing and statistical analysis of microbiome , 2017, Genes & diseases.

[23]  H. Magalon,et al.  Reevaluating species number, distribution and endemism of the coral genus Pocillopora Lamarck, 1816 using species delimitation methods and microsatellites. , 2017, Molecular phylogenetics and evolution.

[24]  R. Peixoto,et al.  Beneficial Microorganisms for Corals (BMC): Proposed Mechanisms for Coral Health and Resilience , 2017, Front. Microbiol..

[25]  A. Schramm,et al.  Endozoicomonas Are Specific, Facultative Symbionts of Sea Squirts , 2016, Front. Microbiol..

[26]  D. Bourne,et al.  Differential specificity between closely related corals and abundant Endozoicomonas endosymbionts across global scales , 2016, The ISME Journal.

[27]  Paul J. McMurdie,et al.  DADA2: High resolution sample inference from Illumina amplicon data , 2016, Nature Methods.

[28]  P. Frade,et al.  The microbiome of coral surface mucus has a key role in mediating holobiont health and survival upon disturbance , 2016, The ISME Journal.

[29]  C. Castro,et al.  Impact of oil spills on coral reefs can be reduced by bioremediation using probiotic microbiota , 2015, Scientific Reports.

[30]  V. Paul,et al.  Microbiome shifts and the inhibition of quorum sensing by Black Band Disease cyanobacteria , 2015, The ISME Journal.

[31]  R. Parsons,et al.  Minor revision to V4 region SSU rRNA 806R gene primer greatly increases detection of SAR11 bacterioplankton , 2015 .

[32]  Ruth D Gates,et al.  The coral core microbiome identifies rare bacterial taxa as ubiquitous endosymbionts , 2015, The ISME Journal.

[33]  Javier Palarea-Albaladejo,et al.  zCompositions — R package for multivariate imputation of left-censored data under a compositional approach , 2015 .

[34]  J. Bythell,et al.  Microbial Communities Associated with Healthy and White Syndrome-Affected Echinopora lamellosa in Aquaria and Experimental Treatment with the Antibiotic Ampicillin , 2015, PloS one.

[35]  René Augustin,et al.  Bacteria–bacteria interactions within the microbiota of the ancestral metazoan Hydra contribute to fungal resistance , 2014, The ISME Journal.

[36]  V. Weis,et al.  TIR-domain-containing protein repertoire of nine anthozoan species reveals coral-specific expansions and uncharacterized proteins. , 2014, Developmental and comparative immunology.

[37]  J. Bythell,et al.  Experimental antibiotic treatment identifies potential pathogens of white band disease in the endangered Caribbean coral Acropora cervicornis , 2014, Proceedings of the Royal Society B: Biological Sciences.

[38]  R. Hill,et al.  Bacterial diversity associated with the tunic of the model chordate Ciona intestinalis , 2013, The ISME Journal.

[39]  D. Bourne,et al.  Nutrient cycling in early coral life stages: Pocillopora damicornis larvae provide their algal symbiont (Symbiodinium) with nitrogen acquired from bacterial associates , 2013 .

[40]  E. Sampayo,et al.  Blind to morphology: genetics identifies several widespread ecologically common species and few endemics among Indo‐Pacific cauliflower corals (Pocillopora, Scleractinia) , 2013 .

[41]  Wei Shi,et al.  featureCounts: an efficient general purpose program for assigning sequence reads to genomic features , 2013, Bioinform..

[42]  Susan Holmes,et al.  phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data , 2013, PloS one.

[43]  A. Kostic,et al.  Exploring host-microbiota interactions in animal models and humans. , 2013, Genes & development.

[44]  D. Oh,et al.  Thalassospiramide G, a New γ-Amino-Acid-Bearing Peptide from the Marine Bacterium Thalassospira sp , 2013, Marine drugs.

[45]  K. Katoh,et al.  MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability , 2013, Molecular biology and evolution.

[46]  Pelin Yilmaz,et al.  The SILVA ribosomal RNA gene database project: improved data processing and web-based tools , 2012, Nucleic Acids Res..

[47]  Paramvir S. Dehal,et al.  FastTree 2 – Approximately Maximum-Likelihood Trees for Large Alignments , 2010, PloS one.

[48]  Emmanuel Paradis,et al.  pegas: an R package for population genetics with an integrated-modular approach , 2010, Bioinform..

[49]  M. Martindale,et al.  The onset of microbial associations in the coral Pocillopora meandrina , 2009, The ISME Journal.

[50]  A. Couloux,et al.  Patterns of genetic structure among Hawaiian corals of the genus Pocillopora yield clusters of individuals that are compatible with morphology. , 2008, Comptes rendus biologies.

[51]  S. Tillier,et al.  The mitochondrial genome of Pocillopora (Cnidaria: Scleractinia) contains two variable regions: the putative D-loop and a novel ORF of unknown function. , 2007, Gene.

[52]  Robert C. Edgar,et al.  MUSCLE: multiple sequence alignment with high accuracy and high throughput. , 2004, Nucleic acids research.

[53]  N. Andreakis,et al.  With eyes wide open: a revision of species within and closely related to the Pocillopora damicornis species complex (Scleractinia; Pocilloporidae) using morphology and genetics , 2014 .

[54]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[55]  P. Gibbs,et al.  Practical applications of contaminant-free Symbiodinium cultures grown on solid media , 2009 .