Abstract
Aliarcobacter butzleri (formerly known as Arcobacter butzleri) is an emerging food-borne zoonotic pathogen that establishes in vitro endosymbiotic relationships with Acanthamoeba castellanii, a free-living amoeba. Previously, we described that this bacterium acts as an endocytobiont of A. castellanii, surviving for at least 10 days in absence of bacterial replication. Thus, the aim of this study was to evaluate the ability of A. butzleri to survive as a long-term endosymbiont of A. castellanii for 30 days in two models of symbiotic interaction with A. castellanii: (i) endosymbiotic culture followed by gentamicin protection assay and (ii) transwell co-culture assay. The results allow us to conclude that A. butzleri is capable of surviving as an endosymbiont of A. castellanii for at least 30 days, without multiplying, under controlled laboratory conditions. In addition, in the absence of nutrients and as both microorganisms remain in the same culture, separated by semi-permeable membranes, A. castellanii does not promote the survival of A. butzleri, nor does it multiply. Our findings suggest that the greater survival capacity of A. butzleri is associated with their endosymbiont status inside A. castellanii, pointing out the complexity of this type of symbiotic relationship.
This is a preview of subscription content, log in via an institution to check access.
References
Almsherqi Z, Hyde S, Ramachandran M, Deng Y (2008) Cubic membranes: a structure-based design for DNA uptake. J R Soc Interface 5:1023–1029. https://doi.org/10.1098/rsif.2007.1351
Anderson I, Watkins RF, Samuelson J et al (2005) Gene discovery in the genome. Protist 156:203–214. https://doi.org/10.1016/j.protis.2005.04.001
Balczun C, Scheid PL (2017) Free-living amoebae as hosts for and vectors of intracellular microorganisms with public health significance. Viruses. https://doi.org/10.3390/v9040065
Burdíková Z, Čapek M, Ostašov P et al (2010) Testate amoebae examined by confocal and two-photon microscopy: implications for taxonomy and ecophysiology. Microsc Microanal 16:735–746. https://doi.org/10.1017/S1431927610094031
Chen CY, Nace GW, Irwin PL (2003) A 6x6 drop plate method for simultaneous colony counting and MPN enumeration of Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli. J Microbiol Methods 55:475–479. https://doi.org/10.1016/S0167-7012(03)00194-5
Collado L, Figueras MJ (2011) Taxonomy, epidemiology, and clinical relevance of the genus Arcobacter. Clin Microbiol Rev 24:174–192. https://doi.org/10.1128/CMR.00034-10
Fera MT, Maugeri TL, Gugliandolo C et al (2008) Induction and resuscitation of viable nonculturable Arcobacter butzleri cells. Appl Environ Microbiol 74:3266–3268. https://doi.org/10.1128/AEM.00059-08
Fernández H, Villanueva MP, Medina G (2012) Endosymbiosis of Arcobacter butzleri in Acanthamoeba castellanii. Rev Argent Microbiol 44:133
Ferreira S, Queiroz JA, Oleastro M, Domingues FC (2015) Insights in the pathogenesis and resistance of Arcobacter: a review. Crit Rev Microbiol. https://doi.org/10.3109/1040841X.2014.954523
Garcia-Sanchez AM, Ariza C, Ubeda JM et al (2013) Free-living amoebae in sediments from the Lascaux Cave in France. Int J Speleol 42:9–13. https://doi.org/10.5038/1827-806X.42.1.2
Gilson PR, Yu X-C, Hereld D et al (2003) Two Dictyostelium orthologs of the prokaryotic cell division protein FtsZ localize to mitochondria and are required for the maintenance of normal mitochondrial morphology. Eukaryot Cell 2:1315–1326. https://doi.org/10.1128/EC.2.6.1315-1326.2003
Greub G, Raoult D (2004) Microorganisms resistant to free-living amoebae. Clin Microbiol Rev 17:413–433
International Commission for the Microbiological Specifications of Foods (2002) Microorganisms in foods 7—microbiological testing in food safety management, 1st edn. Springer, New York
Jung SY, Matin A, Kim KS, Khan NA (2007) The capsule plays an important role in Escherichia coli K1 interactions with Acanthamoeba. Int J Parasitol 37:417–423. https://doi.org/10.1016/j.ijpara.2006.10.012
Kakley MR, Velle KB, Fritz-Laylin LK (2018) Relative quantitation of polymerized actin in suspension cells by flow cytometry. Bio-Protoc. https://doi.org/10.21769/BioProtoc.3094
Kebbi-Beghdadi C, Greub G (2014) Importance of amoebae as a tool to isolate amoeba-resisting microorganisms and for their ecology and evolution: the Chlamydia paradigm. Environ Microbiol Rep 4:309–24
Khan NA, Siddiqui R (2014) Predator vs aliens: bacteria interactions with Acanthamoeba. Parasitology 141:869–874. https://doi.org/10.1017/S003118201300231X
Laskowski-Arce MA, Orth K (2008) Acanthamoeba castellanii promotes the survival of Vibrio parahaemolyticus. Appl Environ Microbiol 74:7183–7188. https://doi.org/10.1128/AEM.01332-08
Medina G, Flores-Martin S, Fonseca B et al (2014) Mechanisms associated with phagocytosis of Arcobacter butzleri by Acanthamoeba castellanii. Parasitol Res 113:1933–1942. https://doi.org/10.1007/s00436-014-3842-8
Medina G, Neves P, Flores-Martin S et al (2019) Transcriptional analysis of flagellar and putative virulence genes of Arcobacter butzleri as an endocytobiont of Acanthamoeba castellanii. Arch Microbiol. https://doi.org/10.1007/s00203-019-01678-0
Mungroo M, Siddiqui R, Khan N (2021) War of the microbial world: Acanthamoeba spp. interactions with microorganisms. Folia Microbiol 66:689–699. https://doi.org/10.1007/s12223-021-00889-7
Mella C, Medina G, Toledo Z (2016) Interaction between zoonotic bacteria and free living amoebas. A new angle of an epidemiological polyhedron of public health importance. Austral J Vet Sci 10:1–10
Molofsky AB, Swanson MS (2004) Differentiate to thrive: Lessons from the Legionella pneumophila life cycle. Mol Microbiol 53:29–40
Oliver JD (2005) The viable but nonculturable state in bacteria. J Microbiol 43((spec 1)):93–100
Oren A, Garrity GM (2014) List of new names and new combinations previously effectively, but not validly, published. Int J Syst Evol Microbiol 64:1–5. https://doi.org/10.1099/ijs.0.060285-0
Pizarro-Cerdá J, Méresse S, Parton RG et al (1998) Brucella abortus transits through the autophagic pathway and replicates in the endoplasmic reticulum of nonprofessional phagocytes. Infect Immun 66:5711–5724
Ramees TP, Dhama K, Karthik K et al (2017) Arcobacter : an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control—a comprehensive review. Vet Q 37:136–161. https://doi.org/10.1080/01652176.2017.1323355
Roy CR, Tilney LG (2002) The road less traveled: transport of Legionella to the endoplasmic reticulum. J Cell Biol 158:415–419. https://doi.org/10.1083/jcb.200205011
Scheid P (2014) Relevance of free-living amoebae as hosts for phylogenetically diverse microorganisms. Parasitol Res 113:2407–2417
Schuster FL (2002) Cultivation of pathogenic and opportunistic free-living amebas. Clin Microbiol Rev 15:342–354. https://doi.org/10.1128/cmr.15.3.342-354.2002
Tsaousis AD, Nývltová E, Šuták R et al (2014) A nonmitochondrial hydrogen production in Naegleria gruberi. Genome Biol Evol 6:792–799. https://doi.org/10.1093/gbe/evu065
Vandamme P, Vancanneyt M, Pot B et al (1992) Polyphasic taxonomic study of the emended genus Arcobacter with Arcobacter butzleri comb. nov. and Arcobacter skirrowii sp. nov., an aerotolerant bacterium isolated from veterinary specimens. Int J Syst Bacteriol 42:344–356. https://doi.org/10.1099/00207713-42-3-344
Villanueva MP, Medina G, Fernández H (2016) Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii. Rev Argent Microbiol. https://doi.org/10.1016/j.ram.2015.12.003
Wojtkowska M, Buczek D, Stobienia O et al (2015) The TOM complex of Amoebozoans: the cases of the amoeba Acanthamoeba castellanii and the slime mold Dictyostelium discoideum. Protist 166:349–362. https://doi.org/10.1016/J.PROTIS.2015.05.005
Yousuf F, Siddiqui R, Khan N (2013) Acanthamoeba castellanii of the T4 genotype is a potential environmental host for Enterobacter aerogenes and Aeromonas hydrophila. Parasit Vectors 6:169. https://doi.org/10.1186/1756-3305-6-169
Zhang X, Zhuchenko O, Kuspa A, Soldati T (2016) Social amoebae trap and kill bacteria by casting DNA nets. Nat Commun 7:10938. https://doi.org/10.1038/ncomms10938
Acknowledgements
I would like to express my deep gratitude to Dra. Carola Otth, my co-sponsor professor who recently pass away, for their patient guidance, enthusiastic encouragement and useful critiques of this paper and the research behind.
Funding
This work was supported by Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT, #1110202), Fondo Concurso Interno Línea Profondecyt Vicerrectoría de Investigación y Postgrado Universidad Católica De Temuco (VIPUCT, #2016PF-GM-03) and Fondo de Equipamiento (FEQUIP2019-CS-05) Vicerrectoría de Investigación y Posgrado Universidad Católica de Temuco.
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Medina, G.A., Flores-Martin, S.N., Pereira, W.A. et al. Long-term survive of Aliarcobacter butzleri in two models symbiotic interaction with Acanthamoeba castellanii. Arch Microbiol 204, 610 (2022). https://doi.org/10.1007/s00203-022-03223-y
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-03223-y