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Cellular Immune Response of an Endemic Lake Baikal Amphipod to Indigenous Pseudomonas sp

Marine Biotechnology volume 23, pages 463–471 (2021)Cite this article

Abstract

Studies of invertebrates have shown that the internal environment of crustaceans is not always sterile in normal conditions, and in many species, it can be populated by microorganisms even in the absence of any visible pathological processes in the body. This observation raises the question of whether genetically modified indigenous hemolymph microorganisms can be used for biotechnological purposes inside the crustacean either as local producers of some compounds or as sensors to physiological parameters. In this study, we tested the ability of the bacteria isolated from the hemolymph of the amphipod Eulimnogammarus verrucosus to hide from the cellular immune response of the host as the most important feature for their potential long-term application in vivo. 16S rDNA amplicon sequencing revealed five common bacterial genera in all analyzed samples of the amphipod hemolymph, among which Pseudomonas is most easily subjected to genome modification and, thus, the most prospective for biotechnological application. Cultivation of Pseudomonas gave us a number of strains undoubtedly derived from the amphipod hemolymph, and one of them (belonging to the Pseudomonas fluorescens group) was chosen for further tests. The primary culture of amphipod hemocytes was used to analyze the immunogenicity of the strain and showed a pronounced reaction of the immune cells to a high amount of the bacteria within six hours. This result indicates that modulation of cellular immune response to metabolically active bacterial cells is not mandatory for the survival and wide distribution of these microorganisms in the hemolymph of numerous amphipod individuals.

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References

  • Balabanova L, Averianova L, Marchenok M, Son O, Tekutyeva L (2021) Microbial and genetic resources for cobalamin (vitamin B12) biosynthesis: from ecosystems to industrial biotechnology. Int J Mol Sci 22:4522

    CAS  Article  Google Scholar 

  • Bel’kova NL, Parfenova VV, Kostornova TY, Denisova LY, Zaichikov EF (2003) Microbial biodiversity in the water of Lake Baikal. Microbiology 72:203–213

    CAS  Article  Google Scholar 

  • Bojko J, Burgess AL, Baker AG, Orr CH (2020) Invasive non-native crustacean symbionts: diversity and impact. J Invertebr Pathol. https://doi.org/10.1016/j.jip.2020.107482

    Article  PubMed  Google Scholar 

  • Bojko J, Dunn AM, Stebbing PD, van Aerle R, Bacela-Spychalska K, Bean TP, Urrutia A, Stentiford GD (2018) ‘Candidatus Aquirickettsiella gammari’ (Gammaproteobacteria: Legionellales: Coxiellaceae): a bacterial pathogen of the freshwater crustacean Gammarus fossarum (Malacostraca: Amphipoda). J Invert Pathol 156:41–53

    Article  Google Scholar 

  • Bojko J, Ovcharenko M (2019) Pathogens and other symbionts of the Amphipoda: taxonomic diversity and pathological significance. Dis Aquat Org 136:3–36

    CAS  Article  Google Scholar 

  • Chattopadhyay P, Banerjee G, Mukherjee S (2017) Recent trends of modern bacterial insecticides for pest control practice in integrated crop management system. Biotech 7:60

    Google Scholar 

  • Cornet S, Biard C, Moret Y (2009) Variation in immune defense among populations of Gammarus pulex (Crustacea: Amphipoda). Oecologia 159:257–269

    Article  Google Scholar 

  • Desriac F, Le Chevalier P, Brillet B, Leguerinel I, Thuillier B, Paillard C, Fleury Y (2014) Exploring the hologenome concept in marine bivalvia: haemolymph microbiota as a pertinent source of probiotics for aquaculture. FEMS Microbiol Lett 350:107–116

    CAS  Article  Google Scholar 

  • Dimova M, Madyarova E, Gurkov A, Drozdova P, Lubyaga Y, Kondrateva E, Timofeyev M (2018) Genetic diversity of Microsporidia in the circulatory system of endemic amphipods from different locations and depths of ancient Lake Baikal. PeerJ. https://doi.org/10.7717/peerj.5329

    Article  PubMed  PubMed Central  Google Scholar 

  • Duval D, Galinier R, Mouahid G, Toulza E, Allienne JF, Portela J, Gourbal B (2015) A novel bacterial pathogen of Biomphalaria glabrata: a potential weapon for schistosomiasis control? PLoS Negl Trop Dis. https://doi.org/10.1371/journal.pntd.0003489

    Article  PubMed  PubMed Central  Google Scholar 

  • Ejeian F, Etedali P, Mansouri-Tehrani HA, Soozanipour A, Low ZX, Asadnia M, Razmjou A (2018) Biosensors for wastewater monitoring: a review. Biosens Bioelectron 118:66–79

    CAS  Article  Google Scholar 

  • Gurkov A, Borvinskaya E, Shchapova E, Timofeyev M (2018) Restraining small decapods and amphipods for in vivo laboratory studies. Crustaceana 91:517–525

    Article  Google Scholar 

  • Gurkov A, Rivarola-Duarte L, Bedulina D, Casas IF, Michael H, Drozdova P, Nazarova A, Govorukhina E, Timofeyev M, Stadler P, Luckenbach T (2019) Indication of ongoing amphipod speciation in Lake Baikal by genetic structures within endemic species. BMC Evol Biol 19:138

    Article  Google Scholar 

  • Haushalter RW, Groff D, Deutsch S, The L, Chavkin TA, Brunner SF, Keasling JD (2015) Development of an orthogonal fatty acid biosynthesis system in E. coli for oleochemical production. Metab Eng 30:1–6

    CAS  Article  Google Scholar 

  • Jakob L, Axenov-Gribanov D, Gurkov A, Ginzburg M, Bedulina D, Timofeyev M, Luckenbach T, Lucassen M, Sartoris F, Pörtner H (2016) Lake Baikal amphipods under climate change: thermal constraints and ecological consequences. Ecosphere. https://doi.org/10.1002/ecs2.1308

    Article  Google Scholar 

  • Li H (2013) Aligning sequence reads, clone sequences and assembly contigs with BWA-MEM. arXiv preprint arXiv: 1303.3997

  • Maksimov IV, Veselova SV, Nuzhnaya TV, Sarvarova ER, Khairullin RM (2015) Plant growth-promoting bacteria in regulation of plant resistance to stress factors. Russ J Plant Physiol 62:715–726

    CAS  Article  Google Scholar 

  • Mardal M, Bischoff M, Ibáñez M, Ruffing U, Hernández F, Meyer MR (2017) Microbial biotransformation of five pyrrolidinophenone-type psychoactive substances in wastewater and a wastewater isolated Pseudomonas putida strain. Drug Test Anal 9:1522–1536

    CAS  Article  Google Scholar 

  • Mastore M, Arizza V, Manachini B, Brivio MF (2015) Modulation of immune responses of Rhynchophorus ferrugineus (Insecta: Coleoptera) induced by the entomopathogenic nematode Steinernema carpocapsae (Nematoda: Rhabditida). Insect Sci 22:748–760

    CAS  Article  Google Scholar 

  • Murali A, Bhargava A, Wright ES (2018) IDTAXA: a novel approach for accurate taxonomic classification of microbiome sequences. Microbiome 6:1–14

    Article  Google Scholar 

  • Newton RJ, Huse SM, Morrison HG, Peake CS, Sogin ML, McLellan SL (2013) Shifts in the microbial community composition of Gulf Coast beaches following beach oiling. PLoS One. https://doi.org/10.1371/journal.pone.0074265

    Article  PubMed  PubMed Central  Google Scholar 

  • Nikolaidis M, Mossialos D, Oliver SG, Amoutzias GD (2020) Comparative analysis of the core proteomes among the Pseudomonas major evolutionary groups reveals species-specific adaptations for Pseudomonas aeruginosa and Pseudomonas chlororaphis. Diversity 12:289

    CAS  Article  Google Scholar 

  • Okonechnikov K, Golosova O, Fursov M, Ugene Team (2012) Unipro UGENE: a unified bioinformatics toolkit. Bioinformatics 28:1166–1167

    Article  Google Scholar 

  • Parks DH, Chuvochina M, Waite DW, Rinke C, Skarshewski A, Chaumeil PA, Hugenholtz P (2018) A standardized bacterial taxonomy based on genome phylogeny substantially revises the tree of life. Nat Biotechnol 36:996–1004

    CAS  Article  Google Scholar 

  • Robach MC (1978) Effect of potassium sorbate on the growth of Pseudomonas fluorescens. J Food Sci 43:1886–1887

    CAS  Article  Google Scholar 

  • Schmitt P, Rosa RD, Duperthuy M, de Lorgeril J, Bachère E, Destoumieux-Garzón D (2012) The antimicrobial defense of the Pacific oyster, Crassostrea gigas. How diversity may compensate for scarcity in the regulation of resident/pathogenic microflora. Front Microbiol 3:160

  • Shchapova E, Nazarova A, Gurkov A, Borvinskaya E, Rzhechitskiy Y, Dmitriev I, Meglinski I, Timofeyev M (2019) Application of PEG-covered non-biodegradable polyelectrolyte microcapsules in the crustacean circulatory system on the example of the amphipod Eulimnogammarus verrucosus. Polymers 11:1246

    CAS  Article  Google Scholar 

  • Singh K, Ainala SK, Kim Y, Park S (2019) A novel D (−)-lactic acid-inducible promoter regulated by the GntR-family protein D-LldR of Pseudomonas fluorescens. Synth Syst Biotechnol 4:157–164

    Article  Google Scholar 

  • Tindall BJ, Rosselló-Móra R, Busse HJ, Ludwig W, Kämpfer P (2010) Notes on the characterization of prokaryote strains for taxonomic purposes. Int J Syst Evol Microbiol 60:249–266

    CAS  Article  Google Scholar 

  • Wang XW, Wang JX (2015) Crustacean hemolymph microbiota: endemic, tightly controlled, and utilization expectable. Mol Immunol 68:404–411

    Article  Google Scholar 

  • Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703

    CAS  Article  Google Scholar 

  • Wright ES, Yilmaz LS, Noguera DR (2012) DECIPHER, a search-based approach to chimera identification for 16S rRNA sequences. Appl Environ Microbiol 78:717–725

    CAS  Article  Google Scholar 

  • Zhang X, Sun Z, Zhang X, Zhang M, Li S (2018) Hemolymph microbiomes of three aquatic invertebrates as revealed by a new cell extraction method. Appl Environ Microbiol 84:e02824-e2917

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang X, Zhang M, Zheng H, Ye H, Zhang X, Li S (2020) Source of hemolymph microbiota and their roles in the immune system of mud crab. Dev Comp Immunol 102:103470

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We acknowledge Dr. Denis Axenov-Gribanov and Evgenii Protasov for help and advice on microbiological issues, as well as Dr. Polina Drozdova for help in the preparation of the manuscript.

Funding

The study was supported primarily by the Russian Foundation for Basic Research (#19–34-90137) and additionally by the Ministry of Science and Higher Education of Russia (#FZZE‐2020‐0026). AN was financially supported by the program «Mikhail Lomonosov» of the German Academic Exchange Service (DAAD) and the Ministry of Science and Higher Education of Russia (application #2300–21).

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Authors and Affiliations

  1. Irkutsk State University, Irkutsk, Russia

    Ekaterina Shchapova, Anna Nazarova, Uliana Vasilyeva, Anton Gurkov, Andrei Mutin, Renat Adelshin & Maxim Timofeyev

  2. Baikal Research Centre, Irkutsk, Russia

    Anton Gurkov & Maxim Timofeyev

  3. Irkutsk Anti-Plague Research Institute of Siberia and Far East, Irkutsk, Russia

    Alexander Ostyak & Renat Adelshin

  4. Scientific Centre for Family Health and Human Reproduction Problems, Irkutsk, Russia

    Natalia Belkova

Authors
  1. Ekaterina Shchapova
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  2. Anna Nazarova
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  3. Uliana Vasilyeva
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  4. Anton Gurkov
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  5. Alexander Ostyak
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  6. Andrei Mutin
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  7. Renat Adelshin
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  8. Natalia Belkova
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  9. Maxim Timofeyev
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Corresponding author

Correspondence to Maxim Timofeyev.

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Ethics Approval

All experimental procedures with amphipods were conducted in accordance with the EU Directive 2010/63/EU for animal experiments and were approved by the Animal Subjects Research Committee of the Institute of Biology at Irkutsk State University.

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10126_2021_10039_MOESM1_ESM.xlsx

Supplementary file1: Table S1. The full list of the strains isolated from hemolymph and chitin surface of E. verrucosus with data on MALDI-TOF MS identification (XLSX 48 KB)

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Shchapova, E., Nazarova, A., Vasilyeva, U. et al. Cellular Immune Response of an Endemic Lake Baikal Amphipod to Indigenous Pseudomonas sp. Mar Biotechnol 23, 463–471 (2021). https://doi.org/10.1007/s10126-021-10039-2

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  • DOI: https://doi.org/10.1007/s10126-021-10039-2

Keywords

  • Amphipoda
  • Baikal
  • Hemolymph
  • Microbiome
  • Microorganisms
  • Symbiosis