Event Abstract

Back to Event

A novel technique for improving an in vitro culture of Demodex spp (Acari: Demodicidae). A pilot trial.

Humberto J. Maldonado Santana1, Dary L. Mendoza Meza2, Luis J. Escaf Jaraba3, Luis C. Escaf3, Martha Lizarazo3, Gloria Garavito De Egea1 and Eduardo Egea Bermejo1*
  • 1 Universidad del Norte, Medicina, Colombia
  • 2 Universidad del Atlantico, Quimica, Colombia
  • 3 Fundaciòn Oftalmologica del Caribe, Oftalmologia, Colombia

Background Demodex folliculorum (Simon, 1842) and Demodex brevis (Akbulatova, 1963) are obligate parasites in humans, Its live in the sebaceous glands and hair follicles sucking nutrients from the hair roots and damaging cell walls. Infestation with Demodex spp has been associated with ocular diseases, such as chronic blepharitis, chalazia or, less commonly, keratoconjunctivitis. The search for agents intended to control the density of Demodex spp requires the establishment and the maintenance of and in vitro culture of this mites. . The optimization of an in vitro or ex vivo demodex spp culture model is necessary for future development of effective treatments Objective The objective of this study was to establish and maintain a novel experimental conditions for an in vitro culture of Demodex spp. Materials and methods Mites Mites were collected by two techniques: First one set up the cellophane tape method. Demodex ssp. were obtained from the face skin of undergraduate volunteer students, aged 18–23, , and the other technique was collecting mites from eyelashes from adult patients with blepharitis referred by ophthalmologist. Demodex folliculorum and Demodex brevis were identified by phenotypic characteristics. Experiment design In order to evaluate the effect of temperature and culture on the long lifespan and viability of the mites, we performed experiments including two variables. . The design was run in a single block and the order of the experiments was completely randomized. The results were analyzed using the Statistical Package SPSS v.22 Temperature and relative humidity were controlled in bioclimatic chamber. Temperature tested were 15, 20 and 25 °C. Range of temperature was fixed to 90%. The culture media tested were: RPMI 1640 with L-glutamine and Hepes buffer. The RPMI medium was used in three combinations: RPMI enriched with human serum, RPMI enriched with fetal bovine serum and plain RPMI. The viability of the mites was observed at intervals of 8 hours, recording the time of death according to criteria of mobility of the body and its appendices, for 1 minute. Results Results indicate that the best alternative to grow and maintain viable mites was in RPMI enriched with human serum .Using this modality we found that mite mortality after 8 days of culture was 40% at 15 ° C and 70% at 25 ° C. Conclusion This is the first description, reporting an in vitro culture of these mites in RPMI with better viability after 8 days of culture compared with those reported in the literature. These results provide an option for the in vitro maintenance of Demodex spp. New trials are in progress to optimize these results.

References

1. Chen 1985. Influence of some environmental factors on the survival of two human follicular mites. Acta Entomological Sinica, Vol. 28, No. 1
2. Shufang J, Lijuan D, Tiongiing L, 2003. Primary studies on conditions for the survival of Demodex folliculorum in vitro. Chinese Journal
3. Zhao Y, Guo N, Zheng X, Yang S, Zhang M, Zhang L, Wu K, 2005. Observations on morphology and the survival temperature range of Demodex folliculorum. Acta Entomological Sinica, Vol 48 (5): 754 – 758.
4. Zhao Y, Guo N, Ping wu L, 2009. The effect of temperature on the viability of Demodex folliculorum and Demodex brevis. Parastitol Res vol 105: 1623-1628.
5. Zhao Y, Guo N, Ping wu L, 2011. Influence of temperature and medium on viability of Demodex folliculorum and Demodex brevis (Acari: Demodicidae). Exp Appl Acarol 54: 421-425.

Keywords: Demodex, culture in vitro, RPMI, Fetal bovine serum, human serum

Conference: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.

Presentation Type: Poster Presentation

Topic: Allergy

Citation: Maldonado Santana HJ, Mendoza Meza DL, Escaf Jaraba LJ, Escaf LC, Lizarazo M, Garavito De Egea G and Egea Bermejo E (2015). A novel technique for improving an in vitro culture of Demodex spp (Acari: Demodicidae). A pilot trial.. Front. Immunol. Conference Abstract: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología. doi: 10.3389/conf.fimmu.2015.05.00265

Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.

The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated.

Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed.

For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions.

Received: 26 May 2015; Published Online: 14 Sep 2015.

* Correspondence: MD, PhD. Eduardo Egea Bermejo, Universidad del Norte, Medicina, Barranquilla, Atlantico, Colombia, egea.eduardo@gmail.com