Abstract
We present a novel and simple technique for storing live Acanthamoeba for long periods of time. The amoebae are maintained at refrigerator temperatures in a peptone–yeast extract–glucose (PYG) medium normally used for cultivation. Using this method, we obtained survival rates of at least 4 years for Acanthamoeba polyphaga and 3 years for Acanthamoeba castellanii and Acanthamoeba rhysodes. Advantages of this storage method are: (1) it is quick and simple, (2) inexpensive, (3) does not require encystment before storage, (4) resuscitation of cysts can be achieved within a week of culture in PYG medium at 27°C, and does not require co-culture with bacteria or any special equipment.
References
Ahearn DG, Gabriel MM (1997) Contact lenses, disinfectants, and Acanthamoeba keratitis. Adv Appl Microbiol 43:35–56
Axelsson-Olsson D, Waldenstrom J, Broman T, Olsen B, Holmberg M (2005) Protozoan Acanthamoeba polyphaga as a potential reservoir for Campylobacter jejuni. Appl Environ Microbiol 71:987–992
Axelsson-Olsson D, Ellstrom P, Waldenstrom J, Haemig PD, Brudin L, Olsen B (2007) Acanthamoeba–Campylobacter coculture as a novel method for enrichment of Campylobacter species. Appl Environ Microbiol 73:6864–6869
Biddick CJ, Rogers LH, Brown TJ (1984) Viability of pathogenic and nonpathogenic free-living amoebae in long-term storage at a range of temperatures. Appl Environ Microbiol 48:859–860
Borazjani RN, May LL, Noble JA, Avery SV, Ahearn DG (2000) Flow cytometry for determination of the efficacy of contact lens disinfecting solutions against Acanthamoeba spp. Appl Environ Microbiol 66:1057–1061
Greub G, Raoult D (2002) Crescent bodies of Parachlamydia acanthamoeba and its life cycle within Acanthamoeba polyphaga: an electron micrograph study. Appl Environ Microbiol 68:3076–3084
Greub G, Raoult D (2004) Microorganisms resistant to free-living amoebae. Clin Microbiol Rev 17:413–433
John DT, Eddy PL, John RA (1994) Cryopreservation of pathogenic free-living amebae. Folia Parasitol (Praha) 41:110–114
Khan NA (2006) Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev 30:564–595
Kilvington S (1990) Activity of water biocide chemicals and contact lens disinfectants on pathogenic free-living amoebae. Int Biodeter 26:127–138
Kilvington S, Price J (1990) Survival of Legionella pneumophila within cysts of Acanthamoeba polyphaga following chlorine exposure. J Appl Bacteriol 68:519–525
Lorenzo-Morales J, Maciver SK (2006) Long-term, low-maintenance storage of Acanthamoeba strains. Parasitol Res 99:743–745
Menrath P, Brame R-M, Oger C, Delattre J-M (1995) A new method of preservation of axenic amoebae of the genera Naegleria and Acanthamoeba. Eur J Protozool 31(1):109–118
Rowbotham TJ (1983) Isolation of Legionella pneumophila from clinical specimens via amoebae, and the interaction of those and other isolates with amoebae. J Clin Pathol 36:978–986
Schuster FL (2002) Cultivation of pathogenic and opportunistic free-living amebas. Clin Microbiol Rev 15(3):342–354
Acknowledgements
This work was supported financially by the Royal Swedish Academy of Agriculture and Forestry (KSLA-H482), the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (FORMAS 221-2007-438), and Sparbankstiftelsen Kronan. All experiments comply with the national guidelines.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Axelsson-Olsson, D., Olofsson, J., Ellström, P. et al. A simple method for long-term storage of Acanthamoeba species. Parasitol Res 104, 935–937 (2009). https://doi.org/10.1007/s00436-008-1304-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-008-1304-x