Hindawi Publishing Corporation BioMed Research International Volume 2013, Article ID 675145, 8 pages http://dx.doi.org/10.1155/2013/675145 Research Article Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey Kübra AçJkalJn CoGkun, 1 Semra Özçelik, 1 Lütfi Tutar, 2 Nazif ElaldJ, 3 and Yusuf Tutar 4,5 1 Department of Parasitology, Faculty of Medicine, Cumhuriyet University, 58140 Sivas, Turkey 2 Department of Biology, Faculty of Science and Letters, Kahramanmaras ¸S¨ utc ¸¨ u ˙ Imam University, 46100 Kahramanmaras, Turkey 3 Department of Infectious Diseases, Faculty of Medicine, Cumhuriyet University, 58140 Sivas, Turkey 4 Department of Biochemistry, Faculty of Pharmacology, Cumhuriyet University, 58140 Sivas, Turkey 5 CUTFAM Research Center, Faculty of Medicine, Cumhuriyet University, 58140 Sivas, Turkey Correspondence should be addressed to Yusuf Tutar; ytutar@cumhuriyet.edu.tr Received 9 April 2013; Revised 11 June 2013; Accepted 27 June 2013 Academic Editor: Gernot Zissel Copyright © 2013 K¨ ubra Ac ¸ıkalın Cos ¸kun et al. Tis 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. Te present work focuses on a local survey of free-living amoebae (FLA) that cause opportunistic and nonopportunistic infections in humans. Determining the prevalence of FLA in water sources can shine a light on the need to prevent FLA related illnesses. A total of 150 samples of tap water were collected from six districts of Sivas province. Te samples were fltered and seeded on nonnutrient agar containing Escherichia coli spread. Tirty-three (22%) out of 150 samples were found to be positive for FLA. Te FLA were identifed by morphology and by PCR using 18S rDNA gene. Te morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of three diferent species, Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. Naegleria fowleri, Balamuthia mandrillaris, or Sappinia sp. was not isolated during the study. All A. castellanii and A. polyphaga sequence types were found to be genotype T4 that contains most of the pathogenic Acanthamoeba strains. Te results indicated the occurrence and distribution of FLA species in tap water in these localities of Sivas, Turkey. Furthermore, the presence of temperature tolerant Acanthamoeba genotype T4 in tap water in the region must be taken into account for health risks. 1. Introduction Free-living amoebae (FLA), ubiquitous and widely dis- tributed protozoa, feed on bacteria, algae, fungi, and small organic particles and are adaptable to their environment [1]. Tey can be found in dust, air, seawater, dental treatment units, sewage, eyewash solutions, contact lenses, and dialysis units and are particularly abundant in soil and water [2, 3]. Among them, only four genera including Acanthamoeba, Naegleria, Balamuthia, and Sappinia cause opportunistic and nonopportunistic infections in humans and in animals, but infections are not commonly reported with the exception of Acanthamoeba keratitis which is reported in over 1 to 2 cases per million contact lens wearers in the USA annually [4– 6]. Acanthamoeba spp. and Balamuthia mandrillaris cause granulomatous amoebic encephalitis (GAE) while Naegleria fowleri causes primary amoebic meningoencephalitis (PAM). Both GAE and PAM are central nervous system infec- tions. Some Acanthamoeba spp., commonly Acanthamoeba castellanii, cause amoebickeratitis (AK), a vision-threatening corneal infection. In humans Acanthamoeba spp. may also afect the skin and lungs [3, 7]. Hartmannella spp. invade animal tissues and have been found in nasal mucosa of humans, the bronchial system of dogs, and the intestines of turkeys [8]. Sappinia diploidea have been reported, only once, from a brain infection in a healthy man [9]. Tis amoeba was identifed later as Sappinia pedata, by using real-time PCR tests based on 18S rRNA gene sequences [10]. Te presence of FLA in tap water may represent a health risk to both immunocompromised and immunocom- petent individuals [11] and they are resistant to extreme conditions of temperature, pH, and exposure to various