جداسازی آمیب نگلریا و شناسایی نگلریا آسترالینسیس از آب استخرهای شنای استان کرمان

نویسندگان
راحله افتخاری کنزرکی 1
کاوس صلح جو 1
زهرا بابایی 2
حسن رضانژاد 3
احمد ابوالقاضی 1
1 گروه انگل شناسی و قارچ شناسی پزشکی، دانشکده پزشکی،دانشگاه علوم پزشکی جهرم، جهرم، ایران
2 مرکز تحقیقات لیشمانیوز، دانشکده پزشکی، دانشگاه علوم پزشکی کرمان، کرمان، ایران
3 مرکز تحقیقات بیماریهای زئونوز، دانشکده پزشکی، دانشگاه علوم پزشکی جهرم، جهرم، ایران
10.52547/jmj.18.2.19
چکیده
مقدمه: در بین آمیب های آزادزی، گونه های N.fowleri و N.australiensis علت مرگ ناشی از مننگوآنسفالیت آمیبی اولیه به حساب می‌آیند. از آن جا که آب استخرها منبع آمیب‌های آزادزی است، این مطالعه با هدف جداسازی آمیب نگلریا و شناسایی نگلریا آسترالینسیس از آب استخرهای شنای استان کرمان انجام شد.
روش کار: 80 نمونه آب از 20  استخر استان کرمان در تابستان 1397 جمع‌آوری و میزان کلر باقی‌مانده، pH و دمای آن‌ها اندازه‌گیری شد. نمونه‌های آب فیلتر و کشت داده شدند. پس از شناسایی مورفولوژیکی آمیب، تعیین جنس به روش  PCR و تایید گونه با مقایسه توالی محصول PCR در بانک ژن انجام شد.
یافته‌ها: دما و باقیمانده کلر به ترتیب در 50% و 93.7 %  از استخرها به میزان استاندارد بود. به طورکلی 8.75 % از نمونه آب استخرها، آلوده به آمیب با مورفولوژی نگلریا بودند. نتیجه PCR حاکی از وجود نگلریا در هفت نمونه بود. درصد آلودگی در سانس آقایان و در یک متری کناره استخر  بیشتر بود. توالی محصول PCR درصد بالایی از هم پوشانی و شباهت با توالی های ثبت شده N.australiensis نشان داد که در بانک ژنی NCBI با شماره MT292609 به ثبت رسید.
نتیجه‌گیری: نتایج پژوهش حاضر نشان داد که آب 25% از استخرهای شنا در استان کرمان به نگلریا آسترالینسیس آلوده هستند. با توجه به توانایی بیماری‌زایی این گونه آمیب در انسان و حیوان، لازم است تمهیدات لازم برای ضدعفونی هر چه بیشتر آب استخرهای شنا توسط دست‌اندرکاران مسائل بهداشتی برای جلوگیری از آلودگی احتمالی اتخاذ شود.
کلیدواژه‌ها
آمیب‌های آزادزی
نگلریا آسترالینسیس
استخر شنا
کرمان

عنوان مقاله English

Detection of Naegleria and Identification of Naegleria australiensis from Swimming pools in Kerman Province

نویسندگان English

Raheleh Eftekhari-Kenzerki 1
Kavous Solhjoo 1
Zahra Babaei 2
Hassan Rezanezhad 3
Ahmad Abolghazi 1
1 Department of Medical Parasitology and Mycology, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran
2 Leishmaniasis Research Center, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran
3 Zoonoses Research Center, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran
چکیده English

Introduction: Among the free living amoeba, Naegleria fowleri and Naegleria australiensis are considered as the cause of death due to primary amoebic meningoencephalitis (PAM). As the water of swimming pools are the sources of free living amoeba, this study conducted to detection of Naegleria and identification of N.australiensis from swimming pools.
Materials and Methods: Eighty water samples were collected from 20 swimming pools of Kerman province in summer 2018 and the residual chlorine, pH and temperature were measured, then filtered and cultured. After morphological identification of amoeba, genus identification and species confirmation were performed by PCR and comparison of PCR product sequence in gene bank, respectively.
Results: The temperature and residual chlorine of water were at standard level in 50% and 93.7%, respectively. Overall 8.75% of water swimming pool samples were contaminated with amoeba with similar morphology to Naegleria. The results of PCR were confirmed the present of Naegleria in 7 samples. The contamination percent was higher in men timetable (10%) and in one meter of wall of swimming pools (12.5%). Comparison of PCR products showed the high percentages of Identity and Query Coverage with recorded sequences of N.australiensis and it was deposited in NCBI with accession number MT292609.
Conclusion: The present research showed that 25% of swimming pools in Kerman province are contaminated with
N. australiensis. Regarding to the pathogenic potential of N. australiensis for human and animals, it is necessary, the health workers think about for more disinfection of swimming pools water for preventing of possible infection.

کلیدواژه‌ها English

Free living amoeba
Naegleria australiensis
Swimming pool
Kerman
1. Scheikl U, Sommer R, Kirschner A, Rameder A, Schrammel B, Zweimüller I, etal. Free-living amoebae (FLA) co-occurring with legionellae in industrial waters. Eur J Protistol. 2014;50(4):422–9 2. World Health Organization. Guidelines for Safe Recreational Water, Coastal and Fresh Waters WHO, Geneva, Switzerland. 2003; Vol. 1, 3. Visvesvara, GS, Moura, H. & Schuster, F. L. Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. FEMS Immunol Med Microbiol 2007; 50 (1), 1–26. 4. Carter, R. F. Description of a Naegleria sp. Isolated from two cases of primary amoebic meningoencephalitis, and the experimental pathological changes induced by it. J. Pathol. 1970; 100:217-244. 5. Ferrante A. Experimental pneumonitis induced by Naegleria fowleri in mice. Trans R Soc Trop Med Hyg. 1981; 75(6):907-8. 6. De Jonckheere, J. F. Naegleria australiensis sp. nov., another pathogenic Naegleria from water. Protistologica 1981; 17:423-429. 7. Visvesvara GS, Sriram R, Qvarnstrom Y et al. Paravahlkampfia francinae n. sp. masquerading as an agent of primary amoebic meningoencephalitis. J Eukaryot Microbiol. 2009; 56(4):357-66. 8. Michel, R., and J. De Jonckheere. Erster Nachweiseiner pathogenen Naegleria-Art (N. australiensis De Jonckheere 1981) in Deutschland. Z. Parasitenkd. 1983; 69:395-396. 9. Scaglia M, Gatti S, Cevini C et al. Naegleria australiensis ssp. italica: experimental study in mice. Exp Parasitol. 1989; 69(3):294-9. 10. Visvesvara GS. Infections with free-living amebae. Handb Clin Neurol. 2013; 114:153-68. 11. De Jonckheere JF, Aerts M, Martinez AJ. Naegleria australiensis: experimental meningoencephalitis in mice. Trans R Soc Trop Med Hyg. 1983; 77(5):712-6. 12. De Jonckheere JF. What do we know by now about the genus Naegleria? Exp Parasitol. 2014; 145 Suppl: S2-9. 13. Movahedi Z, Shokrollahi MR, Aghaali M, Heydari H. Primary amoebic meningoencephalitis in an Iranian infant. Case Rep Med. 2012; 2012:782854. 14. 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):523-9. 15. Solgi R, Niyyati M, Haghighi A, Mojarad EN. Occurrence of Thermotolerant Hartmannella vermiformis and Naegleria Spp. in Hot Springs of Ardebil Province, Northwest Iran. Iran J Parasitol. 2012; 7(2):47-52. 16. 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. 17. Latifi AR, Niyyati M, Lorenzo-Morales J et al. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran. Acta Parasitol. 2017; 62(1):104-109. 18. Latifi A, Niyyati M, Tabaei SJ, Biderouni FT, Haghighi A, Lasjerdi Z. An Experimental Model of Primary Amoebic Meningoencephalitis Due to Naegleria australiensis in Iran. Iran J Parasitol. 2018 Jul;13(3):369. 19. Solgi R., Niyyati M., Haghighi A., Taghipour N., Tabaei S.J., Eftekhar M., et al. Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in northwestern Iran. J Water and Health, 2012; 10:650–656. DOI: 10.2166/wh.2012.032 20. Page F.C. 1988. A new key to freshwater and soil gymnamoebae: with instructions for culture. Ambleside, Cumbria, England, pp 92–96. 21. Pe ́landakis M, Serre S, Pernin P (2000) Analysis of the 5.8S rRNA gene and the internal transcribed spacers in Naegleria spp. and in N. fowleri. J Eukaryot Microbiol, 47: 116–121. 22. Caumo K, Rott MB. Acanthamoeba T3, T4 and T5 in swimming-pool waters from Southern Brazil. Acta tropica. 2011 Mar 1; 117(3):233-5. 23. Seyfried PL, Fraser DJ. Persistence of Pseudomonas areuginosa in chlorinatd swimming pools. Can J Microbiol, 1980; 26(3): 350-5. 24. Rasti S, Assadi MA, Iranshahi L, Saffari M, Gilasi HR, Pourbabaee M. Assessment of microbial contamination and physicochemical condition of public swimming pools in Kashan, Iran. Jundishapour J Microbiol, (2012), 5: 450-455. DOI: 10.5812/jjm.2478 25. Ithoi I, Lau YL, Fadzlun AA, Foead AI, Neilson RS, Nissapatorn V. Detection of free living amoebae, Acanthamoeba and Naegleria, in swimming pools, Malaysia. Tropical Biomedicine. 2010;27(3):566-77. 26. Scaglia M, Strosselli M, Grazioli V, Gatti S, Bernuzzi AM, De Jonckheere JF. Isolation and identification of pathogenic Naegleria australiensis (Amoebida, Vahlkampfiidae) from a spa in northern Italy. Appl Environ Microbiol. 1983 Dec 1;46(6):1282-5. 27. Niyyati M, Saberi R, Lorenzo-Morales J, Salehi R. High occurrence of potentially-pathogenic free-living amoebae in tap water and recreational water sources in South-West Iran. Trop Biomed. 2016 Mar 1;33(1):95-101. 28. Latifi AR, Niyyati M, Lorenzo-Morales J, Haghighi A, Tabaei SJ, Lasjerdi Z, Azargashb E. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran. Acta Parasitologica. 2017 Jan 1;62(1):104-9. 29. Niyyati M, Lasjerdi Z, Nazar M, Haghighi A, Nazemalhosseini Mojarad E. Screening of recreational areas of rivers for potentially pathogenic free-living amoebae in the suburbs of Tehran, Iran. J water and health. 2012 Mar;10(1):140-6. 30. Solgi R, Niyyati M, Haghighi A, Mojarad EN. Occurrence of thermotolerant Hartmannella vermiformis and Naegleria spp. in hot springs of Ardebil Province, Northwest Iran. Iran J Parasitol. 2012;7(2):47. 31. Feiz Haddad MH, Habibpour H, Mahmoudi MR. Isolation and molecular identification of free-living amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) from mineral springs in Guilan Province, northern Iran. J Water and Health. 2020 Feb;18(1):60-6. 32. Milanez G, Masangkay F, Somsak V, Kotepui M, Tangpong J, Karanis P. Occurrence and the first report of Naegleria australiensis presence in a major lake in the Philippines. J water and health. 2019 Aug 1;17(4):647-53.