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2013, Number 3

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Rev Cubana Med Trop 2013; 65 (3)

Preservation of fungal cultures of medical importance in distilled water

Fernández ACM, Díaz SLA, Illnait ZMT, Aragonés LC, Martínez MG, Perurena LMR, Rodríguez GI

Language: Spanish
References: 20
Page: 361-369
PDF size: 73.86 Kb.


ABSTRACT

Introduction: causal agents of the main human myicoses have been preserved in the culture collection of pathogenic fungi at "Pedro KourÍ" Tropical Medicine Institute, a national reference center, with the purpose of using them for medical microbiology diagnoses, research and teaching. Preservation of fungal cultures in sterile distilled water, or Castellani's method, has been endorsed as a method ensuring high rates of strain viability, purity and stability, low costs and great simplicity. It is therefore an advantageous alternative for the maintenance of microorganisms in many laboratories.
Objective: present the results obtained from the preservation in sterile distilled water of the main fungal species included in the collection.
Methods: an evaluation was conducted of the viability, purity and stability of the main morphological and physiological characteristics of 240 strains of different fungal species from the collection, which had been preserved in sterile distilled water.
Results: 80 % of the cultures had retained their viable status without any contamination. The best results corresponded to the preservation of fungi producing abundant conidia (97 % recovery) and yeasts (83 %), followed by dermatophytes (69 %) and dimorphic fungi (68 %). In 8 genera, recovery of viable strains was 100 %, and in 17 it was above 60%. Preservation time was 15-20 years. A web-based digital database was created for the collection.
Conclusions: Castellani's is the method of choice for the preservation of fungal cultures in resource-limited laboratories.


REFERENCES

  1. Smith D. Culture collections over the world. Int Microbiol. 2003;6:95-100.

  2. Hawksworth DL. Fungal diversity and its implications for genetic resources collections. Stud Mycol. 2004;50:9-18.

  3. Nakasone KK, Peterson SW, Jong SC. Preservation and distribution of fungal cultures. En: Mueller GM, Bills GF, Foster MS, editors. Biodiversity of fungi. Inventory and monitoring methods. Amsterdam: Elsevier Academic Press; 2004. p. 37-47.

  4. Hartung de Capriles C, Mata S, Middelveen M. Preservation of fungi in water (Castellani): 20 years. Mycopathologia. 1989;106:73-9.

  5. Diogo HC, Sarpieri A, Pires MC. Preservação de fungos em água destilada. An Bras Dermatol. 2005;80:591-4.

  6. Borman AM, Szekely A, Campbell CK, Johnson EM. Evaluation of the viability of pathogenic filamentous fungi after prolonged storage in sterile water and review of recent published studies on storage methods. Mycopathologia. 2006;161:361-8.

  7. De Hoog GS, Guarro J, Gené J, Figueras MJ. Atlas of clinical fungi. 3rd ed (versión piloto in CD Rom). Utrech¬/Reus: Centraalbureau voor Schimmelcultures/ Universitat Rovira i Virgili; 2009. p. 10-1.

  8. Mendoza M, Alvarado P, Díaz de Torres E, Lucena L, Albornoz MC. Comportamiento fisiológico y de sensibilidad in vitro de aislamientos de Sporothrix schenckii mantenidos 18 años por dos métodos de preservación. Rev Iberoam Micol. 2005;22:151-6.

  9. Qiangqiang Z, Jiajun W, Li L. Storage of fungi using sterile distilled water or lyophilisation: comparison after 12 years. Mycoses. 1998;41:255-7.

  10. Rodrigues EG, Lírio VS, Lacaz CS. Preservação de fungos e actinomicetos de interesse médico em água destilada. Rev Inst Med Trop São Paulo. 1992;34:159-65.

  11. Bueno L, Gallardo R. preservación de hongos filamentosos en agua destilada estéril. Rev Iberoam Micol. 1998;15:166-8.

  12. Stevens D. Clinical aspergillosis for basic scientists. Med Mycol. 2009;47(Suppl. 1):1-4.

  13. Smith D, Onions AH. The preservation and maintenance of living fungi. 2nd ed. Walling Ford: CAB International; 1994.

  14. Lima RF, Borba CM. Viability, morphological characteristics and dimorphic ability of fungi preserved by different methods. Rev Iberoam Micol. 2001;18:191-6.

  15. Pasarell L, Mc Ginnis MR. Viability of fungal cultures maintained at 70 ºC. J Clin Microbiol. 1992;30:1000-4.

  16. Pérez C, Mata-Essayag S, Hartung de Capriles C, Roselló A, Colella MT, Olaizola C, et al. Mantenimiento de Cryptococcus sp. con el método de Castellani. Rev Soc Ven Microbiol. 2003;23:153-7.

  17. Fernández Andreu CM, Díaz Suárez LA, Illnait Zaragozí MT, Aragonés López C, Martínez Machín G, Perurena Lancha MR. Conservación de cultivos fúngicos de alto riesgo de Histoplasma y Cryptococcus. Rev Cubana Med Trop. 2012;64(1):49-54.

  18. Freitas RS, Dantas KC, Pereira CN, Levi JE, Costa Martins JE. Preservation methods of fungi in 35 years old stock culture storages: a comparative study. Afr J Microbiol Res. 2011;5(5):555-61.

  19. García MD, Uruburu F. La conservación de cepas microbianas. Actual Sem. 2000;30:12-6.

  20. Laboratory Procedures for Microorganisms. Establishment and operation of culture collections. Documentation, catalogues and on-line information [citado 12 Nov 2009]. Disponible en: http://www.cabri.org/guidelines/micro-organisms/M101.html




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