Öz
Amaç: Acanthamoeba spp. serbest yaşayan amiplerden olup çevrede deniz sularında, göllerde, nehirlerde, durgun sularda, yüzme havuzlarında, şişelenmiş sularda, havalandırma kanallarında, klima ünitelerinde, lağım sularında, toprakta, hastanelerin diyaliz üniteleri ve göz yıkama üniteleri gibi çok çeşitli yerlerde bulunabilmektedir. Acanthamoeba spp.’ye bağlı enfeksiyonlara az rastlanmasınarağmen yüksek ölüm ile karakterize olup ciddi klinik problemler oluşturmaktadır. Acanthamoeba sağlıklı bireylerde ya da kontakt lens kullanıcılarında Acanthamoeba keratitine (AK) neden olmaktadır. Bağışıklık sistemi baskılanmış kişilerde granülomatöz amebik ensefalit (GAE), akciğer ve deri enfeksiyonlarına sebep olmaktadır. Çalışmamızda Türkiye’nin çeşitli şehirlerinden toplanan çevresel su örneklerinde, Acanthamoeba spp. varlığının farklı kültür ve moleküler yöntem ile araştırılması amaçlanmıştır.
Gereç ve Yöntem: Türkiye’nin farklı şehirlerinden ve su kaynaklarından toplam 100 örnek toplanmıştır. Su örnekleri kültür yöntemi ve DNA izolasyonu için 0.45 μm çapında selüloz nitrat membran filtreden yaklaşık 30 saniyede 100 ml geçecek şekilde vakum cihazı ile filtrelenmiştir. Filtrelenen su örnekleri NNA-E.coli ve Tamponlu Kömür Maya Özütü besiyerlerine kültüre edilmiştir. Kültür sonucu pozitif olarak kabul edilen örnekler DNA izolasyonu için -20 °C dondurucuya kaldırılmıştır.
Bulgular: Toplanan 100 örneğin kültür sonuçları besiyeri ayrımı yapılmaksızın 27 (%27) Acanthamoeba spp. tespit edilmiştir. NNA-E. coli besiyerinde 22 (%22) BCYE besiyerinde 19 (%19) Acanthamoeba spp. üretilmiştir. İki besiyeri arasında anlamlı bir farklılık bulunamamıştır. RT-PCR yöntemi ile toplam 25 Acanthamoeba spp. saptanmıştır (%25). Besiyerlerinin RT-PCR ile uyumu kappa katsayısı hesaplanarak istatiksel olarak (p<0,005) anlamlı ve iki yönlü olarak kabul edilebilir bulunmuştur. Acanthamoeba spp. kisti saptanan örnekler morfolojik kriterlere göre değerlendirilmiştir. Kistlerin büyüklük ve şekilleri baz alınarak yapılan sınıflamada ağırlıklı olarak tespit edilen örneklerin grup II ve grup III’e ait oldukları görülmüştür.
Sonuç: Acanthamoeba büyümesini farklı kültür ortamlarında karşılaştıran az sayıda çalışma yapılmıştır. Bu amip için daha yeni ve daha erişilebilir kültür ortamları keşfedilmesine ihtiyaç vardır.
Anahtar Kelimeler
Acanthamoeba spp. çevresel su örnekleri real time PCR kültür
Destekleyen Kurum
İstanbul Üniversitesi Bilimsel Araştırma Projeleri Birimi
Proje Numarası
38220
Kaynakça
- Castrillón JC, Orozco LP. Acanthamoeba spp. como parásitos patógenos y oportunistas. Revista Chil Infectol 2013;30(2):147-55. [CrossRef] google scholar
- Schuster FL, Visvesvara GS. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol 2004;34:1001-27. [CrossRef] google scholar
- Center for Disease Control and Prevention (CDC). Acanthamoeba Infections, Acanthamoeba Keratitis. 2024 September. https://www.cdc.gov/acanthamoeba/about/ about-acanthamoeba-keratitis.html google scholar
- Satlin MJ, Graham JK, Visvesvara GS, Mena H, Marks KM, Saal SD, et al. Fulminant and fatal encephalitis caused by Acanthamoeba in a kidney transplant recipient: case report and literature review. Transpl Infect Dis 2013;15(6):619-26. [CrossRef] google scholar
- Putaporntip C, Kuamsab N, Nuprasert W, Rojrung R, Pattanawong U, Tia T, et al. Analysis of Acanthamoeba genotypes from public freshwater sources in Thailand reveals a new genotype, T23 Acanthamoeba bangkokensis sp. nov. Sci Rep 2021;11(1):17290. [CrossRef] google scholar
- Raju R, Khurana S, Mahadevan A, John DV. Central nervous system infections caused by pathogenic free-living amoebae: An Indian perspective. Trop Biomed 2022;39(2):265-80. [CrossRef] google scholar
- Penland RL, Wilhelmus KR. Comparison of axenic and monoxenic media for isolation of Acanthamoeba. J Clin Microbiol 1997;35(4):915-22. [CrossRef] google scholar
- Tawfeek GM, Bishara SAH, Sarhan RM, Taher EE, Khayyal AE. Genotypic, physiological, and biochemical characterization of potentially pathogenic Acanthamoeba isolated from the environment in Cairo, Egypt. Parasitol Res 2016;115(5):1871-81. [CrossRef] google scholar
- Mahmoudi MR, Maleki M, Zebardast N, Rahmati B, Ashrafi K, Sharifdini M, et al. Identification of Acanthamoeba spp. from water and soil of public parks in the north of Iran. J Water Health 2022(10);1604-10. [CrossRef] google scholar
- Hajialilo E, Behnia M, Tarighi F, Niyyati M, Rezaeian M. Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran. Parasitol Res 2016;115(8):3147-51. [CrossRef] google scholar
- Al-Herrawy AZ, Heshmat MG, Abu Kabsha, SH, Gad MA, Lotfy WM. Occurrence of Acanthamoeba species in the Damanhour drinking water treatment plant, Behera Governorate (Egypt). Reports in Parasitology 2015;4:15-21. [CrossRef] google scholar
- Milanez GD, Masangkay FR, Scheid P, Dionisio JD, Somsak V, Kotepui M, et al. Acanthamoeba species isolated from Philippine freshwater systems: epidemiological and molecular aspects. Parasitol Res 2020;119(11):3755-61. [CrossRef] google scholar
- Karimi A, Kheirandish F, Mamaghani AJ, Taghipour N, Mousavi SF, Aghajani A, et al. Identification and genotyping of Acanthamoeba spp. in the water resources of western Iran. Parasite Epidemiol Control 2023;22:e00308. [CrossRef] google scholar
- Kao PM, Chou MY, Tao CW, Huang WC, Hsu BM, Shen SM, et al. Diversity and seasonal impact of Acanthamoeba species in a subtropical rivershed. BioMed Res Int 2013;2013:405794. [CrossRef] google scholar
INVESTIGATION OF ACANTHAMOEBA SPP. WITH CULTURE AND MOLECULAR METHODS IN THE ENVIRONMENTAL WATER SAMPLES
Öz
Objective: Acanthamoeba spp. are free-living amoebae found in a variety of environments, including seawater, lakes, rivers, stagnant waters, swimming pools, bottled waters, ventilation ducts, air conditioning units, sewage systems, soil, and in-hospital dialysis and eye wash units. Although infections caused by Acanthamoeba spp. are infrequent, they are characterised by high mortality and can lead to serious clinical problems. Acanthamoeba causes Acanthamoeba keratitis (AK) in healthy individuals and contact lens users. In immunocompromised individuals, it can lead to granulomatous amoebic encephalitis (GAE) and lung and skin infections. In this study, we investigated the presence of Acanthamoeba species in environmental water samples collected from various cities across Turkey. We employed a range of culture and molecular methods for this analysis.
Material and Methods: A total of 100 samples were collected from different cities and water sources in Turkey. Water samples were filtered through a 0.45 μm diameter cellulose nitrate membrane filter using a vacuum device, allowing approximately 100 ml to pass in about 30 s. The filtered water samples were cultured on Non-Nutrient Agar E.coli (NNA-E.coli) and Buffered Charcoal Yeast Extract Agar media. Samples that were considered culture-positive were stored at -20 °C for DNA isolation.
Results: Among the 100 samples, 27 (27%) Acanthamoeba spp. were detected without differentiation by the media. In the NNA-E. coli medium, 22 (22%) Acanthamoeba spp. were produced, while 19 (19%) were produced in the BCYE medium. No significant difference was found between the two media. A total of 25 Acanthamoeba spp. were detected using RT-PCR (25%). The compatibility of the media with RT-PCR was found to be statistically significant (p<0.005) and bidirectional, as calculated by the kappa coefficient. Samples in which Acanthamoeba spp. cysts were detected were evaluated based on morphological criteria. Based on the size and shape of the cysts, it was observed that the majority of the detected samples belonged to groups II and III.
Conclusion: There are few studies comparing Acanthamoeba growth in different culture media. There is a need to discover newer and more accessible culture media for this amoeba.
Anahtar Kelimeler
Acanthamoeba spp. environment water samples real time PCR culture
Destekleyen Kurum
Istanbul University Scientific Research Projects Unit
Proje Numarası
38220
Kaynakça
- Castrillón JC, Orozco LP. Acanthamoeba spp. como parásitos patógenos y oportunistas. Revista Chil Infectol 2013;30(2):147-55. [CrossRef] google scholar
- Schuster FL, Visvesvara GS. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol 2004;34:1001-27. [CrossRef] google scholar
- Center for Disease Control and Prevention (CDC). Acanthamoeba Infections, Acanthamoeba Keratitis. 2024 September. https://www.cdc.gov/acanthamoeba/about/ about-acanthamoeba-keratitis.html google scholar
- Satlin MJ, Graham JK, Visvesvara GS, Mena H, Marks KM, Saal SD, et al. Fulminant and fatal encephalitis caused by Acanthamoeba in a kidney transplant recipient: case report and literature review. Transpl Infect Dis 2013;15(6):619-26. [CrossRef] google scholar
- Putaporntip C, Kuamsab N, Nuprasert W, Rojrung R, Pattanawong U, Tia T, et al. Analysis of Acanthamoeba genotypes from public freshwater sources in Thailand reveals a new genotype, T23 Acanthamoeba bangkokensis sp. nov. Sci Rep 2021;11(1):17290. [CrossRef] google scholar
- Raju R, Khurana S, Mahadevan A, John DV. Central nervous system infections caused by pathogenic free-living amoebae: An Indian perspective. Trop Biomed 2022;39(2):265-80. [CrossRef] google scholar
- Penland RL, Wilhelmus KR. Comparison of axenic and monoxenic media for isolation of Acanthamoeba. J Clin Microbiol 1997;35(4):915-22. [CrossRef] google scholar
- Tawfeek GM, Bishara SAH, Sarhan RM, Taher EE, Khayyal AE. Genotypic, physiological, and biochemical characterization of potentially pathogenic Acanthamoeba isolated from the environment in Cairo, Egypt. Parasitol Res 2016;115(5):1871-81. [CrossRef] google scholar
- Mahmoudi MR, Maleki M, Zebardast N, Rahmati B, Ashrafi K, Sharifdini M, et al. Identification of Acanthamoeba spp. from water and soil of public parks in the north of Iran. J Water Health 2022(10);1604-10. [CrossRef] google scholar
- Hajialilo E, Behnia M, Tarighi F, Niyyati M, Rezaeian M. Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran. Parasitol Res 2016;115(8):3147-51. [CrossRef] google scholar
- Al-Herrawy AZ, Heshmat MG, Abu Kabsha, SH, Gad MA, Lotfy WM. Occurrence of Acanthamoeba species in the Damanhour drinking water treatment plant, Behera Governorate (Egypt). Reports in Parasitology 2015;4:15-21. [CrossRef] google scholar
- Milanez GD, Masangkay FR, Scheid P, Dionisio JD, Somsak V, Kotepui M, et al. Acanthamoeba species isolated from Philippine freshwater systems: epidemiological and molecular aspects. Parasitol Res 2020;119(11):3755-61. [CrossRef] google scholar
- Karimi A, Kheirandish F, Mamaghani AJ, Taghipour N, Mousavi SF, Aghajani A, et al. Identification and genotyping of Acanthamoeba spp. in the water resources of western Iran. Parasite Epidemiol Control 2023;22:e00308. [CrossRef] google scholar
- Kao PM, Chou MY, Tao CW, Huang WC, Hsu BM, Shen SM, et al. Diversity and seasonal impact of Acanthamoeba species in a subtropical rivershed. BioMed Res Int 2013;2013:405794. [CrossRef] google scholar
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Sağlık Hizmetleri ve Sistemleri (Diğer) |
| Bölüm | ARAŞTIRMA |
| Yazarlar | |
| Proje Numarası | 38220 |
| Yayımlanma Tarihi | 28 Nisan 2025 |
| Gönderilme Tarihi | 12 Kasım 2024 |
| Kabul Tarihi | 18 Mart 2025 |
| Yayımlandığı Sayı | Yıl 2025 Cilt: 88 Sayı: 2 |
Kaynak Göster
Contact information and address
Addressi: İ.Ü. İstanbul Tıp Fakültesi Dekanlığı, Turgut Özal Cad. 34093 Çapa, Fatih, İstanbul, TÜRKİYE
Email: itfdergisi@istanbul.edu.tr
Phone: +90 212 414 21 61