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Journal of Parasitology Research
Volume 2014, Article ID 256310, 10 pages
http://dx.doi.org/10.1155/2014/256310
Research Article

Morphological Features and In Vitro Cytopathic Effect of Acanthamoeba griffini Trophozoites Isolated from a Clinical Case

1Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, CINVESTAV-IPN, Avenida Instituto Politécnico Nacional 2508, San Pedro Zacatenco, 07360 Mexico, DF, Mexico
2Faculty of Superior Studies, Biology, UNAM, Los Reyes Iztacala, 54090 Tlalnepantla, MEX, Mexico
3University Institute of Tropical Diseases and Public Health of The Canary Islands, University of La Laguna, Avenida Astrofísico Francisco Sánchez SN, San Cristóbal de La Laguna, Tenerife, 38203 Canary Islands, Spain
4Centre for Integrative Physiology, School of Biomedical Sciences, University of Edinburgh, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK

Received 3 June 2014; Revised 14 August 2014; Accepted 18 August 2014; Published 8 September 2014

Academic Editor: Renato A. Mortara

Copyright © 2014 Arturo González-Robles et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Light and transmission electron microscopy observations are reported on the structure and in vitro cytopathic effect of Acanthamoeba griffini trophozoites isolated from a clinical case. Live trophozoites were moderately active with a remarkable pleomorphism which changed from ovoid to quite elongated shapes. When moving, amoebae formed cytoplasmic projections such as wide lamellae and acanthopodia of diverse size and thickness which contain a significant amount of actin. Ultrastructurally, the cytoplasm showed the main organelles found in other free-living amoebae. Coincubation of trophozoites with MDCK cell monolayers resulted in a local damage to target cells after 24 h of interaction, suggesting that the cytopathic effect is contact-dependent. By transmission electron microscopy, amoebae appeared to engulf small portions of the MDCK cells; however, the cells that were not in contact with trophozoites had an unaltered morphology. When epithelial monolayers were incubated with conditioned medium for 24 h, small areas of cell injury were also observed. The phylogenetical analysis as well as the sequencing of the acquired amplified product for the DF3 region of the amoebae isolate confirmed that it belongs to genotype T3, which includes other pathogenic amoebae; besides the activity of two drugs currently used against Acanthamoeba was tested on A. griffini.