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      Detection of Acanthamoeba and Toxoplasma in River Water Samples by Molecular Methods in Iran

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          Abstract

          Background:

          Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP) method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples.

          Methods:

          A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively.

          Results:

          Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites.

          Conclusion:

          The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oo)cystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran.

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          Most cited references33

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          Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue.

          Motoki Goto, Eiichi Honda, Atsuo Ogura … (2009)
          Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after adding an intercalating dye to the reaction solution, but the use of such dyes has certain limitations. Hydroxy naphthol blue (HNB), a metal indicator for calcium and a colorimetric reagent for alkaline earth metal ions, was used for a new colorimetric assay of the LAMP reaction. Preaddition of 120 microM HNB to the LAMP reaction solution did not inhibit amplification efficiency. A positive reaction is indicated by a color change from violet to sky blue. The LAMP reaction with HNB could also be carried out in a 96-well microplate, and the reaction could be measured at 650 nm with a microplate reader. The colorimetric LAMP method using HNB would be helpful for high-throughput DNA and RNA detection.
          Article 0 comments Cited 289 times – based on 0 reviews     Review now
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            Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt.

            Panagiotis Karanis, Christina Kourenti, Huw Smith (2007)
            At least 325 water-associated outbreaks of parasitic protozoan disease have been reported. North American and European outbreaks accounted for 93% of all reports and nearly two-thirds of outbreaks occurred in North America. Over 30% of all outbreaks were documented from Europe, with the UK accounting for 24% of outbreaks, worldwide. Giardia duodenalis and Cryptosporidium parvum account for the majority of outbreaks (132; 40.6% and 165; 50.8%, respectively), Entamoeba histolytica and Cyclospora cayetanensis have been the aetiological agents in nine (2.8%) and six (1.8%) outbreaks, respectively, while Toxoplasma gondii and Isospora belli have been responsible for three outbreaks each (0.9%) and Blastocystis hominis for two outbreaks (0.6%). Balantidium coli, the microsporidia, Acanthamoeba and Naegleria fowleri were responsible for one outbreak, each (0.3%). Their presence in aquatic ecosystems makes it imperative to develop prevention strategies for water and food safety. Human incidence and prevalence-based studies provide baseline data against which risk factors associated with waterborne and foodborne transmission can be identified. Standardized methods are required to maximize public health surveillance, while reporting lessons learned from outbreaks will provide better insight into the public health impact of waterborne pathogenic protozoa.
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              Toxoplasmosis - a waterborne zoonosis.

              J Dubey (2004)
              Humans become infected with Toxoplasma gondii mainly by ingesting uncooked meat containing viable tissue cysts or by ingesting food or water contaminated with oocysts from the feces of infected cats. Circumstantial evidence suggests that oocyst-induced infections in humans are clinically more severe than tissue cyst-acquired infections. Until recently, water-borne transmission of T. gondii was considered uncommon but a large human outbreak linked to contamination of a municipal water reservoir in Canada by wild felids and the widespread infection by marine mammals in the USA provide reasons to question this view. The present paper reviews information on the biology of oocyst-induced infections of T. gondii in humans and animals and examines possible importance of transmission by water.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Iran J Parasitol
                Journal ID (iso-abbrev): Iran J Parasitol
                Journal ID (publisher-id): IJPA
                Journal ID (pmc): IJPA
                Title: Iranian Journal of Parasitology
                Publisher: Tehran University of Medical Sciences
                ISSN (Print): 1735-7020
                ISSN (Electronic): 2008-238X
                Publication date (Print): Apr-Jun 2015
                Volume: 10
                Issue: 2
                Pages: 250-257
                Affiliations
                [1. ]Dept. of Medical Microbiology and Parasitology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran
                [2. ]Dept. of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
                [3. ]Cellular and Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran
                [4. ]Center for Biomedicine and Infectious Diseases (CBID), Qinghai Academy for Animal Sciences and Veterinary Medicine, Qinghai University, Xining, China
                Author notes
                [* ] Correspondence Email: mrmahmoodi2002@ 123456yahoo.com
                Article
                Publisher ID: ijpa-10-250
                PMC ID: 4522301
                PubMed ID: 26246823
                SO-VID: 528c8496-5fe8-49b9-bfd0-ac1f1bcbaf9f
                Copyright © Copyright© Iranian Society of Parasitology & Tehran University of Medical Sciences
                License:

                This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.

                History
                Date received : 9 December 2014
                Date accepted : 13 April 2015
                Categories
                Subject: Original Article

                ScienceOpen disciplines: Parasitology
                Keywords: acanthamoeba,toxoplasma,pcr,lamp,iran
                Data availability:
                ScienceOpen disciplines: Parasitology
                Keywords: acanthamoeba, toxoplasma, pcr, lamp, iran

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