Isolation and molecular characterization of potentially pathogenic Acanthamoeba genotypes from diverse water resources including household drinking water from Khyber Pakhtunkhwa, Pakistan
- 435 Downloads
Acanthamoeba, an opportunistic protozoan pathogen, is ubiquitous in nature, and therefore plays a predatory role and helps control microbial communities in the ecosystem. These Acanthamoeba species are recognized as opportunistic human pathogens that may cause blinding keratitis and rare but fatal granulomatous encephalitis. To date, there is not a single report demonstrating Acanthamoeba isolation and identification from environmental sources in Pakistan, and that is the aim of this study. Acanthamoeba were identified by morphological characteristics of their cysts on non-nutrient agar plates seeded with Escherichia coli. Additionally, the polymerase chain reaction (PCR) was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Furthermore, our PCR and sequencing results confirmed seven different pathogenic and nonpathogenic genotypes, including T2–T10, T4, T5, T7, T15, T16, and T17. To the best of our knowledge, we have identified and isolated Acanthamoeba sp., for the first time, from water resources of Khyber Pakhtunkhwa, Pakistan. There is an urgent need to address (1) the pathogenic potential of the identified genotypes and (2) explore other environmental sources from the country to examine the water quality and the current status of Acanthamoeba species in Pakistan, which may be a potential threat for public health across the country.
KeywordsKeratitis Acanthamoeba Keratitis Acanthamoeba Isolation Acanthamoeba Species Single Polymerase Chain Reaction Product
The authors are thankful to Brig. Muhammad Farooq Murawat, Asim Wazir Khan Naizi, Sohail Khan Naizi, Junaid Khan, Abu Turab Khan, and Ghazala Shabnum for their tremendous support in sample collection and transportation. This work was partially supported by grants from Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan.
Conflicts of interest
Authors have no conflict of interest to declare.
- Arnalich-Montiel F, Lorenzo-Morales J, Irigoyen C, Morcillo-Laiz R, López-Vélez R, Muñoz-Negrete F, Piñero JE, Valladares B (2012) Co-isolation of Vahlkampfia and Acanthamoeba in Acanthamoeba-like keratitis in a Spanish population. Cornea 0:1–7Google Scholar
- Booton GC, Kelly DJ, Chu YW, Seal DV, Houang E, Lam DSC, Byers TJ, Fuerst PA (2002) 18S ribosomal DNA typing and tracking of Acanthamoeba sp. isolates from corneal scrape specimens, contact lens, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong. J Clin Microbiol 40:1621–1625PubMedCrossRefGoogle Scholar
- Gregorio D, Rivasi CF, Mongiardo N, De Rienzo B, Visvesvara GS (1991) Acanthamoeba meningoencephalitis in an AIDS patient: first report from Europe. Arch Pathol Lab Med 116:1363–1365Google Scholar
- Lorenzo-Morales J, Ortega-Rivas A, Martínez E, Khoubbane M, Artigas P, Victoria Periago M, Foronda P, Abreu-Acosta N, Valladares B, Mas-Coma S (2006) Acanthamoeba isolates belonging to T1, T2, T3, T4, and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Trop 100:63–69PubMedCrossRefGoogle Scholar
- Niyyati M, Lorenzo-Morales J, Rahimic F, Motevalli-Haghia A, Martín-Navarro CM, Farniaa S, Valladares B, Rezaeiana M (2009) Isolation and genotyping of potentially pathogenic Acanthamoeba strains from dust sources in Iran. Trans R Soc Trop Med Hyg. doi: 10.1016/j.trstmh.2008.12.007 PubMedGoogle Scholar
- Rezaeian M, Niyyati M, Farnia S, Haghi AM (2008) Isolation of Acanthamoeba spp. from different environmental sources. Iran J Parasitol 3:44–47Google Scholar