Isolation and Molecular Identification of Acanthamoeba and Naegleria from Agricultural Water Canal in Qazvin, Iran
,
Abstract
Background: Free-living amoeba (FLA) are widely distributed in different environmental sources. The most genera of the amoeba are Acanthamoeba, Naegleria and Vermamoeba. The most common consequences of the infections in immune-deficient and immunocompetent persons are amoebic encephalitis and keratitis. The aim of this study was to investigate the presence of Acanthamoeba spp. and Naegleria spp., isolated from the main agricultural water canal in Qazvin.
Methods: Totally, 120 water specimens were collected and later the specimens were cultured and cloned to identify positive samples. PCR amplification and sequencing were carried out to identify the isolated species as well as the genotypes of amoeba.
Results: According to morphological surveys, 41.7% (50/120) of water specimens were positive for FLA. Molecular analysis revealed that 68.6% and 31.4% of Acanthamoeba specimens were identified as T3 and T4 genotypes, respectively. Also, two species of Naegleria named as N. lovaniensis (57.1%) and Naegleria spp. (42.8%) were identified. The results of pathogenicity assays demonstrated that 38.5% of T3 and 61.5% of T4 genotypes of Acanthamoeba were highly pathogenic parasites.
Conclusion: The water flowing in the agricultural canal of the area is contaminated with potential pathogenic FLA, therefore, it is recommended that more attention to be paid towards proper treatment of water sources to prevent possible risk of the disease.
1. Schuster FL, Visvesvara GS. Amebae and ciliated protozoa as causal agents of waterborne zoonotic disease. Vet Parasitol. 2004;126(1-2):91-120.
2. Siddiqui R, Khan NA. Biology and pathogenesis of Acanthamoeba. Parasit Vectors. 2012;5:6.
3. Khan NA. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev. 2006;30(4):564-95.
4. Martinez AJ, Visvesvara GS. Free‐living, amphizoic and opportunistic amebas. Brain Pathol. 1997;7(1):583-98.
5. Niyyati M, Rezaeian M. Current status of Acanthamoeba in Iran: a narrative review article. Iran J Parasitol. 2015;10(2):157-63.
6. Lorenzo-Morales J, Martín-Navarro CM, López-Arencibia A, et al. Acanthamoeba keratitis: an emerging disease gathering importance worldwide? Trends Parasitol. 2013;29(4):181-7.
7. Marciano-Cabral F, Cabral GA. The immune response to Naegleria fowleri amebae and pathogenesis of infection. FEMS Immunol Med Microbiol. 2007;51(2):243-59.
8. Visvesvara GS, Moura H, Schuster FL. Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. FEMS Immunol Med Microbiol. 2007;50(1):1-26.
9. Corsaro D, Walochnik J, Köhsler M, et al. Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19, and T20 and proposal for Acanthamoeba micheli sp. nov.(genotype T19). Parasitol Res. 2015;114(7):2481-90.
10. Nuprasert W, Putaporntip C, Pariyakanok L, et al. Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol. 2010;48(12):4636-40.
11. Karakavuk M, Aykur M, Şahar EA, et al. First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target? Exp Parasitol. 2017;183:137-42.
12. Fuerst PA, Booton GC. Species, Sequence Types and Alleles: Dissecting Genetic Variation in Acanthamoeba. Pathogens. 2020;9(7):534.
13. Schuster FL, Visvesvara GS. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol. 2004;34(9):1001-27.
14. Hajialilo E, Behnia M, Tarighi F, et al. Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran. Parasitol Res. 2016;115(8):3147-51.
15. Maghsood AH, Sissons J, Rezaian M, et al. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. J Med Microbiol. 2005;54(8):755-9.
16. Niyyati M, Lorenzo-Morales J, Rezaie S, et al. Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran. Exp Parasitol. 2009;121(3):242-5.
17. Latifi AR, Niyyati M, Lorenzo-Morales J, et al. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran. Acta Parasitologica. 2017;62:104-9.
18. Solgi R, Niyyati M, Haghighi A, et al. Occurrence of thermotolerant Hartmannella vermiformis and Naegleria spp. in hot springs of Ardebil Province, Northwest Iran. Iran J Parasitol. 2012;7(2):47-52.
19. Niyyati M, Lasjerdi Z, Nazar M, et al. Screening of recreational areas of rivers for potentially pathogenic free-living amoebae in the suburbs of Tehran, Iran. J Water Health. 2012;10(1):140-6.
20. Niyyati M, Lasjerdi Z, Zarein-Dolab S, et al. Morphological and molecular survey of Naegleria spp. in water bodies used for recreational purposes in Rasht city, Northern Iran. Iran J Parasitol. 2015;10(4):523-29.
21. Majid MAA, Mahboob T, Mong BGJ, et al. Pathogenic waterborne free-living amoebae: An update from selected Southeast Asian countries. PLoS One. 2017;12(2):e0169448.
22. Khan NA. Pathogenicity, morphology, and differentiation of Acanthamoeba. Curr Microbiol. 2001;43(6):391-5.
23. Memari F, Niyyati M, Haghighi A, et al. Occurrence of pathogenic Acanthamoeba genotypes in nasal swabs of cancer patients in Iran. Parasitol Res. 2015;114(5):1907-12.
24. Anderson OR. A Half-century of Research on Free-living Amoebae (1965-2017): Review of Biogeographic, Ecological and Physiological Studies. Acta Protozoologica. 2018;57(1):1-28.
25. Król-Turmińska K, Olender A. Human infections caused by free-living amoebae. Ann Agric Environ Med. 2017;24(2):254-60.
26. Hooshyar H, Hosseinbigi B, Saraei M, et al. Genotyping of Acanthamoeba isolated from surface and stagnant waters of Qazvin, Central Iran. Iran Red Crescent Med J. 2013;15(6):536-8.
27. Rahdar M, Niyyati M, Salehi M, et al. Isolation and genotyping of Acanthamoeba strains from environmental sources in Ahvaz City, Khuzestan Province, Southern Iran. Iran J Parasitol. 2012;7(4):22-6.
28. Salehi M. Acanthamoeba Strains genotypes prevalence in water Sources in Bojnurd City: Short Communication. J Birjand Univ Med Sci. 2014;21(2):260-6.
29. Eftekhar M, Nazemalhosseini Mojarad E, Haghighi A, et al. Detection of Acanthamoeba from fresh water using polymerase chain reaction. Research in Medicine. 2009;33(1):43-46.
30. Ghadar-ghadr SH, Solhjoo K, Norouz-nejad MJ, et al. Isolation and identification of free living amoeba (Naegleria and Acanthamoeba) in Shiraz water resources by morphological criteria. Pars of Jahrom University of Medical Sciences. 2012;10(3):33-42.
31. Mosayebi M, Ghorbanzadeh B, Eslamirad Z, et al. The Isolation and Detection of Acanthamoeba in Rural Water Sources of Arak, Iran. Mljgoums. 2014;7(4):66-71.
32. Niyyati M, Nazar M, Lasjerdi Z, et al. Reporting of T4 genotype of Acanthamoeba isolates in recreational water sources of Gilan province, northern Iran. Novelty in Biomedicine. 2015;3(1):20-4.
33. Aghajani A, Dabirzadeh M, Maroufi Y, et al. Identification of Acanthamoeba genotypes in pools and stagnant water in ponds in Sistan region in Southeast Iran. Turkiye Parazitol Derg. 2016;40(3):132-136.
34. Ghaderifar S, Najafpoor AA, Zarrinfar H, et al. Isolation and identification of Acanthamoeba from pond water of parks in a tropical and subtropical region in the Middle East, and its relation with physicochemical parameters. BMC Microbiol. 2018;18(1):139.
35. Montalbano Di Filippo M, Santoro M, Lovreglio P, et al. Isolation and molecular characterization of free-living amoebae from different water sources in Italy. Int J Environ Res Public Health. 2015;12(4):3417-27.
36. Lorenzo-Morales J, Ortega-Rivas A, Martínez E, et al. Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Trop. 2006;100(1-2):63-9.
37. Kiss C, Barna Z, Vargha M, et al. Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary. Parasitol Res. 2014;113(7):2551-7.
38. Thammaratana T, Laummaunwai P, Boonmars T. Isolation and identification of Acanthamoeba species from natural water sources in the northeastern part of Thailand. Parasitol Res. 2016;115(4):1705-9.
39. Edagawa A, Kimura A, Kawabuchi-Kurata T, et al. Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan. Parasitol Res. 2009;105(4):1109-17.
40. Lass A, Guerrero M, Li X, et al. Detection of Acanthamoeba spp. in water samples collected from natural water reservoirs, sewages, and pharmaceutical factory drains using LAMP and PCR in China. Sci Total Environ. 2017;584-585:489-94.
41. Coşkun KA, Özçelik S, Tutar L, et al . Isolation and identification of free-living amoebae from tap water in Sivas, Turkey. Biomed Res Int. 2013;2013.
42. De Jonckheere JF. What do we know by now about the genus Naegleria? Exp Parasitol. 2014;145:S2-S9.
43. Najafpoor AA, Zarrinfar H, Ghaderifar S, et al. Naegleria species population found in pond water of parks in Mashhad city, Can the physicochemical factors affect it? MethodsX. 2018;5:1427-30.
44. Javanmard E, Niyyati M, Lorenzo-Morales J, et al. Molecular identification of waterborne free living amoebae (Acanthamoeba, Naegleria and Vermamoeba) isolated from municipal drinking water and environmental sources, Semnan province, north half of Iran. Exp Parasitol. 2017;183:240-4.
45. Değerli S, Değerli N, Çamur D, et al. Genotyping by Sequencing of Acanthamoeba and Naegleria Iso-lates from the Thermal Pool Distributed Through-out Turkey. Acta Parasitologica. 2020;65(1):175-186.
46. Wei W, Feng W, Jiping L, et al. Isolation and identification of Naegleria species from environmental water in changchun, northeastern china. Iran J Parasitol. 2014;9(2):254-9.
47. Nazar M, Haghighi A, Niyyati M, et al. Genotyping of Acanthamoeba isolated from water in recreational areas of Tehran, Iran. J Water Health. 2011;9(3):603-8.
48. Shokri A, Sarvi S, Daryani A, et al. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran. Korean J Parasitol. 2016;54(4):447-53.
49. Behnia M, Hatam-Nahavandi K, Hajialilo E, et al. Occurrence of Acanthamoeba Genotypes in Wastewater Samples in Tehran, Iran. Iran J Parasitol. 2017;12(4):516-21.
50. Niyyati M, Lasgerdi Z, Lorenzo-Morales J. Detection and molecular characterization of potentially pathogenic free-living amoebae from water sources in Kish Island, Southern Iran. Microbiol Insights. 2015;8:1-6.
51. Armand B, Motazedian M, Asgari Q. Isolation and identification of pathogenic free-living amoeba from surface and tap water of Shiraz City using morphological and molecular methods. Parasitol Res. 2016;115(1):63-8.
52. Behniafar H, Niyyati M, Lasjerdi Z. Molecular Characterization of Pathogenic Acanthamoeba Isolated from Drinking and Recreational water in East Azerbaijan, Northwest Iran. Environ Health Insights. 2015;9:7-12.
2. Siddiqui R, Khan NA. Biology and pathogenesis of Acanthamoeba. Parasit Vectors. 2012;5:6.
3. Khan NA. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev. 2006;30(4):564-95.
4. Martinez AJ, Visvesvara GS. Free‐living, amphizoic and opportunistic amebas. Brain Pathol. 1997;7(1):583-98.
5. Niyyati M, Rezaeian M. Current status of Acanthamoeba in Iran: a narrative review article. Iran J Parasitol. 2015;10(2):157-63.
6. Lorenzo-Morales J, Martín-Navarro CM, López-Arencibia A, et al. Acanthamoeba keratitis: an emerging disease gathering importance worldwide? Trends Parasitol. 2013;29(4):181-7.
7. Marciano-Cabral F, Cabral GA. The immune response to Naegleria fowleri amebae and pathogenesis of infection. FEMS Immunol Med Microbiol. 2007;51(2):243-59.
8. Visvesvara GS, Moura H, Schuster FL. Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. FEMS Immunol Med Microbiol. 2007;50(1):1-26.
9. Corsaro D, Walochnik J, Köhsler M, et al. Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19, and T20 and proposal for Acanthamoeba micheli sp. nov.(genotype T19). Parasitol Res. 2015;114(7):2481-90.
10. Nuprasert W, Putaporntip C, Pariyakanok L, et al. Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol. 2010;48(12):4636-40.
11. Karakavuk M, Aykur M, Şahar EA, et al. First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target? Exp Parasitol. 2017;183:137-42.
12. Fuerst PA, Booton GC. Species, Sequence Types and Alleles: Dissecting Genetic Variation in Acanthamoeba. Pathogens. 2020;9(7):534.
13. Schuster FL, Visvesvara GS. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol. 2004;34(9):1001-27.
14. Hajialilo E, Behnia M, Tarighi F, et al. Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran. Parasitol Res. 2016;115(8):3147-51.
15. Maghsood AH, Sissons J, Rezaian M, et al. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. J Med Microbiol. 2005;54(8):755-9.
16. Niyyati M, Lorenzo-Morales J, Rezaie S, et al. Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran. Exp Parasitol. 2009;121(3):242-5.
17. Latifi AR, Niyyati M, Lorenzo-Morales J, et al. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran. Acta Parasitologica. 2017;62:104-9.
18. Solgi R, Niyyati M, Haghighi A, et al. Occurrence of thermotolerant Hartmannella vermiformis and Naegleria spp. in hot springs of Ardebil Province, Northwest Iran. Iran J Parasitol. 2012;7(2):47-52.
19. Niyyati M, Lasjerdi Z, Nazar M, et al. Screening of recreational areas of rivers for potentially pathogenic free-living amoebae in the suburbs of Tehran, Iran. J Water Health. 2012;10(1):140-6.
20. Niyyati M, Lasjerdi Z, Zarein-Dolab S, et al. Morphological and molecular survey of Naegleria spp. in water bodies used for recreational purposes in Rasht city, Northern Iran. Iran J Parasitol. 2015;10(4):523-29.
21. Majid MAA, Mahboob T, Mong BGJ, et al. Pathogenic waterborne free-living amoebae: An update from selected Southeast Asian countries. PLoS One. 2017;12(2):e0169448.
22. Khan NA. Pathogenicity, morphology, and differentiation of Acanthamoeba. Curr Microbiol. 2001;43(6):391-5.
23. Memari F, Niyyati M, Haghighi A, et al. Occurrence of pathogenic Acanthamoeba genotypes in nasal swabs of cancer patients in Iran. Parasitol Res. 2015;114(5):1907-12.
24. Anderson OR. A Half-century of Research on Free-living Amoebae (1965-2017): Review of Biogeographic, Ecological and Physiological Studies. Acta Protozoologica. 2018;57(1):1-28.
25. Król-Turmińska K, Olender A. Human infections caused by free-living amoebae. Ann Agric Environ Med. 2017;24(2):254-60.
26. Hooshyar H, Hosseinbigi B, Saraei M, et al. Genotyping of Acanthamoeba isolated from surface and stagnant waters of Qazvin, Central Iran. Iran Red Crescent Med J. 2013;15(6):536-8.
27. Rahdar M, Niyyati M, Salehi M, et al. Isolation and genotyping of Acanthamoeba strains from environmental sources in Ahvaz City, Khuzestan Province, Southern Iran. Iran J Parasitol. 2012;7(4):22-6.
28. Salehi M. Acanthamoeba Strains genotypes prevalence in water Sources in Bojnurd City: Short Communication. J Birjand Univ Med Sci. 2014;21(2):260-6.
29. Eftekhar M, Nazemalhosseini Mojarad E, Haghighi A, et al. Detection of Acanthamoeba from fresh water using polymerase chain reaction. Research in Medicine. 2009;33(1):43-46.
30. Ghadar-ghadr SH, Solhjoo K, Norouz-nejad MJ, et al. Isolation and identification of free living amoeba (Naegleria and Acanthamoeba) in Shiraz water resources by morphological criteria. Pars of Jahrom University of Medical Sciences. 2012;10(3):33-42.
31. Mosayebi M, Ghorbanzadeh B, Eslamirad Z, et al. The Isolation and Detection of Acanthamoeba in Rural Water Sources of Arak, Iran. Mljgoums. 2014;7(4):66-71.
32. Niyyati M, Nazar M, Lasjerdi Z, et al. Reporting of T4 genotype of Acanthamoeba isolates in recreational water sources of Gilan province, northern Iran. Novelty in Biomedicine. 2015;3(1):20-4.
33. Aghajani A, Dabirzadeh M, Maroufi Y, et al. Identification of Acanthamoeba genotypes in pools and stagnant water in ponds in Sistan region in Southeast Iran. Turkiye Parazitol Derg. 2016;40(3):132-136.
34. Ghaderifar S, Najafpoor AA, Zarrinfar H, et al. Isolation and identification of Acanthamoeba from pond water of parks in a tropical and subtropical region in the Middle East, and its relation with physicochemical parameters. BMC Microbiol. 2018;18(1):139.
35. Montalbano Di Filippo M, Santoro M, Lovreglio P, et al. Isolation and molecular characterization of free-living amoebae from different water sources in Italy. Int J Environ Res Public Health. 2015;12(4):3417-27.
36. Lorenzo-Morales J, Ortega-Rivas A, Martínez E, et al. Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Trop. 2006;100(1-2):63-9.
37. Kiss C, Barna Z, Vargha M, et al. Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary. Parasitol Res. 2014;113(7):2551-7.
38. Thammaratana T, Laummaunwai P, Boonmars T. Isolation and identification of Acanthamoeba species from natural water sources in the northeastern part of Thailand. Parasitol Res. 2016;115(4):1705-9.
39. Edagawa A, Kimura A, Kawabuchi-Kurata T, et al. Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan. Parasitol Res. 2009;105(4):1109-17.
40. Lass A, Guerrero M, Li X, et al. Detection of Acanthamoeba spp. in water samples collected from natural water reservoirs, sewages, and pharmaceutical factory drains using LAMP and PCR in China. Sci Total Environ. 2017;584-585:489-94.
41. Coşkun KA, Özçelik S, Tutar L, et al . Isolation and identification of free-living amoebae from tap water in Sivas, Turkey. Biomed Res Int. 2013;2013.
42. De Jonckheere JF. What do we know by now about the genus Naegleria? Exp Parasitol. 2014;145:S2-S9.
43. Najafpoor AA, Zarrinfar H, Ghaderifar S, et al. Naegleria species population found in pond water of parks in Mashhad city, Can the physicochemical factors affect it? MethodsX. 2018;5:1427-30.
44. Javanmard E, Niyyati M, Lorenzo-Morales J, et al. Molecular identification of waterborne free living amoebae (Acanthamoeba, Naegleria and Vermamoeba) isolated from municipal drinking water and environmental sources, Semnan province, north half of Iran. Exp Parasitol. 2017;183:240-4.
45. Değerli S, Değerli N, Çamur D, et al. Genotyping by Sequencing of Acanthamoeba and Naegleria Iso-lates from the Thermal Pool Distributed Through-out Turkey. Acta Parasitologica. 2020;65(1):175-186.
46. Wei W, Feng W, Jiping L, et al. Isolation and identification of Naegleria species from environmental water in changchun, northeastern china. Iran J Parasitol. 2014;9(2):254-9.
47. Nazar M, Haghighi A, Niyyati M, et al. Genotyping of Acanthamoeba isolated from water in recreational areas of Tehran, Iran. J Water Health. 2011;9(3):603-8.
48. Shokri A, Sarvi S, Daryani A, et al. Isolation and Genotyping of Acanthamoeba spp. as Neglected Parasites in North of Iran. Korean J Parasitol. 2016;54(4):447-53.
49. Behnia M, Hatam-Nahavandi K, Hajialilo E, et al. Occurrence of Acanthamoeba Genotypes in Wastewater Samples in Tehran, Iran. Iran J Parasitol. 2017;12(4):516-21.
50. Niyyati M, Lasgerdi Z, Lorenzo-Morales J. Detection and molecular characterization of potentially pathogenic free-living amoebae from water sources in Kish Island, Southern Iran. Microbiol Insights. 2015;8:1-6.
51. Armand B, Motazedian M, Asgari Q. Isolation and identification of pathogenic free-living amoeba from surface and tap water of Shiraz City using morphological and molecular methods. Parasitol Res. 2016;115(1):63-8.
52. Behniafar H, Niyyati M, Lasjerdi Z. Molecular Characterization of Pathogenic Acanthamoeba Isolated from Drinking and Recreational water in East Azerbaijan, Northwest Iran. Environ Health Insights. 2015;9:7-12.
| Files | ||
| Issue | Vol 15 No 3 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v15i3.4204 | |
| Keywords | ||
| Acanthamoeba Naegleria Genotype Agricultural water Iran | ||
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How to Cite
1.
KHORSANDI RAFSANJANI M, HAJIALILO E, SARAEI M, ALIZADEH SA, JAVADI A. Isolation and Molecular Identification of Acanthamoeba and Naegleria from Agricultural Water Canal in Qazvin, Iran. Iran J Parasitol. 2020;15(3):393-402.
