Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

PLoS One. 2016 Apr 22;11(4):e0154246. doi: 10.1371/journal.pone.0154246. eCollection 2016.

Abstract

Purpose: Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus.

Methods: One hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba.

Results: Among the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of "not cleaning the CL case" presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples.

Conclusions: The PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acanthamoeba / genetics
  • Acanthamoeba / physiology*
  • Acanthamoeba Keratitis / diagnosis
  • Acanthamoeba Keratitis / parasitology*
  • Acanthamoeba Keratitis / prevention & control
  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Child
  • Contact Lens Solutions / analysis*
  • Contact Lenses / microbiology
  • Contact Lenses / parasitology*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification
  • DNA, Protozoan / chemistry
  • DNA, Protozoan / genetics
  • DNA, Protozoan / isolation & purification
  • Disinfection / methods
  • Disinfection / standards
  • Female
  • Host-Parasite Interactions
  • Humans
  • Hygiene / standards
  • Male
  • Middle Aged
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / physiology
  • Risk Factors
  • Sequence Analysis, DNA
  • Spain
  • Staphylococcus aureus / genetics
  • Staphylococcus aureus / physiology
  • Surveys and Questionnaires
  • Young Adult

Substances

  • Contact Lens Solutions
  • DNA, Bacterial
  • DNA, Protozoan

Grants and funding

This project was supported by grant PI12/02725 from Instituto de Salud Carlos III (FISS) (http://www.isciii.es/) and FEDER and by grant USPCEU-PC07/2013 of the Fundación Universitaria San Pablo CEU (http://www.ceu.es/). TSG was supported by Research Program EADS CASA-Brasil I and by grant BEX 9132/13-9 of the Coordenaçao de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)/Brazil (http://www.capes.gov.br/). AM was supported by FPU grant AP2009-0415 of Ministerio de Educación, Cultura y Deporte, Spain (http://www.mecd.gob.es/portada-mecd/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.