Intra-amoebic localization of Arcobacter butzleri as an endocytobiont of Acanthamoeba castellanii
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Intra‑amoebic localization of Arcobacter butzleri as an endocytobiont of Acanthamoeba castellanii G. Medina1 · P. Leyán1 · C. Viera da Silva2 · S. Flores‑Martin3 · C. Manosalva4 · H. Fernández3 Received: 9 May 2019 / Revised: 19 June 2019 / Accepted: 1 July 2019 © Springer-Verlag GmbH Germany, part of Springer Nature 2019
Abstract Acanthamoeba castellanii is a free-living amoeba found mainly in humid environments and Arcobacter butzleri is an emerging zoonotic pathogen, both can establish in vitro endosymbiotic relationships in the absence of bacterial replication. We analyzed the localization of A. butzleri within A. castellanii establishing their association with endoplasmic reticulum vesicles and mitochondria. Through confocal microscopy, we observed that during the early stages of endosymbiosis, there is not colocalization between amoebic vacuoles containing A. butzleri and mitochondria or ER vesicles of A. castellanii. Considering that energy production of this bacterium occurs via metabolism of amino acids or the tricarboxylic acid cycle, these results contribute to explain the absence of bacterial replication, since A. butzleri would not have access to the nutrients found in endoplasmic reticulum vesicles and mitochondria. In addition, we observe that A. butzleri induces significantly the actin polymerization of A. castellanii during the early stages of endosymbiosis. Keywords Acanthamoeba · Arcobacter · Endosymbiosis · Confocal microscopy
Introduction Free-living amoebae (FLA), including Acanthamoeba castellanii, assume a fundamental role in the maintenance of ecosystems, as they feed primarily on bacteria, controlling the biomass of these organisms in the environment (Siddiqui Communicated by Erko Stackebrandt. * G. Medina [email protected]; [email protected] C. Viera da Silva [email protected] S. Flores‑Martin [email protected] 1
Faculty of Health Sciences, Department of Diagnostic Processes and Evaluation, Universidad Católica de Temuco, PO. BOX 15‑D, Temuco, Chile
2
Institute of Biomedical Sciences, Universidade Federal de Uberlândia, Av. Amazonas, Bloco 6T, Campus Umuarama, CEP, Uberlândia, MG 38400‑902, Brazil
3
Institute of Clinical Microbiology,Faculty of Medicine, Universidad Austral de Chile, PO. BOX 567, Valdivia, Chile
4
Institute of Pharmacy, Faculty of Science, Universidad Austral de Chile, Valdivia, Chile
and Khan 2012). Nonetheless, co-culture and metagenomics studies have shown that a wide range of bacterial species, including some of clinical importance for humans and other mammals, are resistant to amoebic phagocytosis, being able to establish endosymbiotic relationships with FLA (Evstigneeva et al. 2009; Garcia-Sanchez et al. 2013; Delafont et al. 2013). Arcobacter butzleri is considered an emerging food-borne zoonotic pathogen worldwide and is the most frequently specie of the genus isolated from environmental water, food, and human clinical samples (Collado and Figueras 2011; Ramees et al. 2017). This pathogen is associated with abortion
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