Metabolomics Study of Immune Responses of New Zealand Greenshell™ Mussels (Perna canaliculus) Infected with Pathogenic Vibrio sp.
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Vibrio coralliilyticus is a bacterial pathogen which can affect a range of marine organisms, such as corals, fish and shellfish, with sometimes devastating consequences. However, little is known about the mechanisms involved in the host-pathogen interaction, especially within molluscan models. We applied gas chromatography-mass spectrometry (GC-MS)-based metabolomics to characterize the physiological responses in haemolymph of New Zealand Greenshell™ mussels (Perna canaliculus) injected with Vibrio sp. DO1 (V. coralliilyticus/neptunius-like isolate). Univariate data analyses of metabolite profiles in Vibrio-exposed mussels revealed significant changes in 22 metabolites at 6 h post-infection, compared to non-exposed mussels. Among them, 10 metabolites were up-regulated, while 12 metabolites were down-regulated in infected mussels. Multivariate analyses showed a clear distinction between infected and non-infected mussels. In addition, secondary pathway analyses indicated perturbations of the host innate immune system following infection, including oxidative stress, inflammation and disruption of the TCA cycle, change in amino acid metabolism and protein synthesis. These findings provide new insights into the pathogenic mechanisms of Vibrio infection of mussels and demonstrate our ability to detect detailed and rapid host responses from haemolymph samples using a metabolomics approach.
KeywordsMussel immunology Metabolite profiles Bacterial infection Host-pathogen interactions Shellfish health Flow cytometry Haemolymph
We would like to thank Westpac Mussel Distributors Ltd. for supplying mussels, Aditya Kesarcodi-Watson (Cawthron Institute, Nelson, New Zealand) for providing the bacterial strain and Erica Zarate and Saras Green (University of Auckland) for their assistance with metabolite sample processing. This project was supported by the New Zealand Ministry of Business, Innovation and Employment (MBIE) (CAWX1315). Additional financial support was provided by a New Zealand ASEAN Scholarship and an award from the New Zealand Marine Sciences Society (NZMSS) to T. V. Nguyen, under the supervision of A. C. Alfaro and F. Merien. We are also thankful to all the members of the Aquaculture Biotechnology Research Group at the Auckland University of Technology (ABRG-AUT) for their assistant during this project.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- Alfaro AC, Young T (2016) Showcasing metabolomic applications in aquaculture: a review. Rev Aquac. https://doi.org/10.1111/raq.12152
- Brown C (1981) A study of two shellfish-pathogenic Vibrio strains isolated from a Long Island hatchery during a recent outbreak of disease [New York]. J Shellfish Res 1:83–87Google Scholar
- Chouchani ET, Pell VR, Gaude E, Aksentijević D, Sundier SY, Robb EL, Logan A, Nadtochiy SM, Ord ENJ, Smith AC, Eyassu F, Shirley R, Hu CH, Dare AJ, James AM, Rogatti S, Hartley RC, Eaton S, Costa ASH, Brookes PS, Davidson SM, Duchen MR, Saeb-Parsy K, Shattock MJ, Robinson AJ, Work LM, Frezza C, Krieg T, Murphy MP (2014) Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS. Nature 515:431–435CrossRefPubMedPubMedCentralGoogle Scholar
- Costa MM, Prado-Alvarez M, Gestal C, Li H, Roch P, Novoa B, Figueras A (2009) Functional and molecular immune response of Mediterranean mussel (Mytilus galloprovincialis) haemocytes against pathogen-associated molecular patterns and bacteria. Fish Shellfish Immunol 26:515–523CrossRefPubMedGoogle Scholar
- Fleury E, Huvet A, Lelong C, de Lorgeril J, Boulo V, Gueguen Y, Bachère E, Tanguy A, Moraga D, Fabioux C, Lindeque P, Shaw J, Reinhardt R, Prunet P, Davey G, Lapègue S, Sauvage C, Corporeau C, Moal J, Gavory F, Wincker P, Moreews F, Klopp C, Mathieu M, Boudry P, Favrel P (2009) Generation and analysis of a 29,745 unique expressed sequence tags from the Pacific oyster (Crassostrea gigas) assembled into a publicly accessible database: the GigasDatabase. BMC Genomics 10:341CrossRefPubMedPubMedCentralGoogle Scholar
- Hasanuzzaman AFM, Robledo D, Gómez-Tato A, Alvarez Dios JA, Harrison PW, Cao A, Fernández-Boo S, Villalba A, Pardo BG, Martínez P (2017) Transcriptomic profile of manila clam (Ruditapes philippinarum) haemocytes in response to Perkinsus olseni infection. Aquaculture 467:170–181CrossRefGoogle Scholar
- Jha AK, Huang SCC, Sergushichev A, Lampropoulou V, Ivanova Y, Loginicheva E, Chmielewski K, Stewart KM, Ashall J, Everts B, Pearce EJ, Driggers EM, Artyomov MN (2015) Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization. Immunity 42:419–430CrossRefPubMedGoogle Scholar
- Kesarcodi-Watson A, Miner P, Nicolas J-L, Robert R (2012) Protective effect of four potential probiotics against pathogen-challenge of the larvae of three bivalves: Pacific oyster (Crassostrea gigas), flat oyster (Ostrea edulis) and scallop (Pecten maximus). Aquaculture 344–349:29–34CrossRefGoogle Scholar
- Lampropoulou V, Sergushichev A, Bambouskova M, Nair S, Vincent EE, Loginicheva E, Cervantes-Barragan L, Ma X, Huang SCC, Griss T, Weinheimer CJ, Khader S, Randolph GJ, Pearce EJ, Jones RG, Diwan A, Diamond MS, Artyomov MN (2016) Itaconate links inhibition of succinate dehydrogenase with macrophage metabolic remodeling and regulation of inflammation. Cell Metab 24:158–166CrossRefPubMedPubMedCentralGoogle Scholar
- Li E, Li C (2014) Use of RNA-seq in aquaculture research. Poult Fish Wildl Sci 2:108–109Google Scholar
- Michelucci A, Cordes T, Ghelfi J, Pailot A, Reiling N, Goldmann O, Binz T, Wegner A, Tallam A, Rausell A, Buttini M, Linster CL, Medina E, Balling R, Hiller K (2013) Immune-responsive gene 1 protein links metabolism to immunity by catalyzing itaconic acid production. Proc Natl Acad Sci 110:7820–7825CrossRefPubMedPubMedCentralGoogle Scholar
- Milan M, Coppe A, Reinhardt R, Cancela LM, Leite RB, Saavedra C, Ciofi C, Chelazzi G, Patarnello T, Bortoluzzi S, Bargelloni L (2011) Transcriptome sequencing and microarray development for the manila clam, Ruditapes philippinarum: genomic tools for environmental monitoring. BMC Genomics 12:234CrossRefPubMedPubMedCentralGoogle Scholar
- Moreira R, Milan M, Balseiro P, Romero A, Babbucci M, Figueras A, Bargelloni L, Novoa B (2014) Gene expression profile analysis of manila clam (Ruditapes philippinarum) hemocytes after a vibrio alginolyticus challenge using an immune-enriched oligo-microarray. BMC Genomics 15:267CrossRefPubMedPubMedCentralGoogle Scholar
- Plumb JA, Hanson LA (2011) Health maintenance and principal microbial diseases of cultured fishes. Wiley, New YorkGoogle Scholar
- Sanders ER (2012) Aseptic laboratory techniques: plating methods. J Vis Exp 63:3064Google Scholar
- Voet D, Voet JG, Pratt CW (2016) Fundamentals of biochemistry: life at the molecular level. Wiley, New YorkGoogle Scholar
- Young T, Alfaro AC (2016) Metabolomic strategies for aquaculture research: a primer. Rev Aquac 0:1–31Google Scholar