Isolation and molecular identification of free-living amoebae from dishcloths in Tenerife, Canary Islands, Spain
In this work, the presence of free-living amoebae (FLA) in dishcloths collected from human activity related places was evaluated. Once in the laboratory, 6 cm2 pieces of each dishcloth were cut and washed with Page’s Amoeba Solution (PAS) in sterile tubes. After washing, the dishcloth pieces were removed, and the tubes were centrifuged (1500 rpm for 10 min). The obtained pellets were seeded onto 2% non-nutrient agar (NNA) plates, incubated at room temperature and were monitored daily an inverted microscope. Once clonal cultures were obtained (only one type of FLA observed), molecular analyses were carried out in order to characterize the isolated FLA strains at the genus/genotype level. From the 31 dishcloths which were processed, FLA strains were isolated in NNA plates in 13 the samples (13/31, 42%). However, and due to bacterial overgrowth, only six strains were characterized at the molecular level (PCR and sequencing). Among the PCR positive strains, 83.33% (5/6) of the PCR positive samples belonged to Acanthamoeba genus (80% (4/5) to genotype T4 and 20% (1/5) to genotype T11). Furthermore, one strain was identified as a member of Allovahlkampfia genus using both morphological and molecular approaches. To the best of our knowledge, this is the first report on the isolation of Allovahlkampfia genus from dishcloths and in the Spanish territory. The presence of FLA in dishcloths should raise awareness to improve hygienic strategies in food- and domestic-related environments, in order to prevent contamination with these protozoa, which are able to be pathogenic and even to act as vehicles of other pathogenic agents.
KeywordsAcanthamoeba Allovahlkampfia PCR Dishcloths Canary Islands Spain
This work was supported by the grants PI18/01380, Fondo Europeo de Desarollo Regional (FEDER) and RICET [project no. RD16/0027/0001 of the programme of Redes Temáticas de Investigación Cooperativa, FIS], Spanish Ministry of Science, Innovation and Universities, Madrid, Spain. MRB was also funded by RICET. ALA and IS were funded by Agustin de Betancourt programme. OC was funded by a grant from Vicerrectorado de Internacionalización, Universidad de La Laguna.
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Conflict of interests
The authors declare that they have no conflict of interest.
- Cabello-Vílchez AM, Mena R, Zuñiga J, Cermeño P, Martín-Navarro CM, González AC, López-Arencibia A, Reyes-Batlle M, Piñero JE, Valladares B, Lorenzo-Morales J (2014) Endosymbiotic Mycobacterium chelonae in a Vermamoeba vermiformis strain isolated from the nasal mucosa of an HIV patient in Lima, Peru. Exp Parasitol 145(Supl):S127–S130CrossRefGoogle Scholar
- Lorenzo-Morales J, Monteverde-Miranda CA, Jiménez C, Tejedor ML, Valladares B, Ortega-Rivas A (2005). Evaluation of Acanthamoeba isolates from environmental sources in Tenerife, Canary Islands, Spain. Ann Agric Environ Med 12(2):233–236Google Scholar
- Lorenzo-Morales J, Morcillo-Laiz R, Martín-Navarro CM, López-Vélez R, López-Arencibia A, Arnalich-Montiel F, Maciver SK, Valladares B, Martínez-Carretero E, Lorenzo-Morales J, Morcillo-Laiz R, Martín-Navarro CM (2011) López-Vélez (2011) Acanthamoeba keratitis due to genotype T11 in a rigid gas permeable contact lens wearer in Spain. Cont Lens Anterior Eye 34(2):83–86. https://doi.org/10.1016/j.clae.2010.10.007. CrossRefGoogle Scholar
- Lorenzo-Morales J, Martín-Navarro CM, López-Arencibia A, Arnalich-Montiel F, Piñero JE, Valladares B (2013) Acanthamoeba keratitis: an emerging disease gathering importance worldwide?. Trends Parasitol 29(4):181–187Google Scholar
- Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16(2):273–307Google Scholar
- Pagnier I, Yutin N, Croce O, Makarova KS, Wolf YI, Benamar S, Raoult D, Koonin EV, La Scola B (2015a) Babela massiliensis, a representative of a widespread bacterial phylum with unusual adaptations to parasitism in amoebae. Biol Direct 31:10–13. https://doi.org/10.1186/s13062-015-0043-z. Google Scholar
- Reyes-Batlle M, Niyyati M, Martín-Navarro CM, López-Arencibia A, Valladares B, Martínez-Carretero E, Piñero JE, Lorenzo-Morales J (2015) Unusual Vermamoeba vermiformis strain isolated from snow in Mount Teide, Tenerife, Canary Islands, Spain. NBM 3:189–192Google Scholar
- Reyes-Batlle M, Girbau C, López-Arencibia A, Sifaoui I, Liendo AR, Bethencourt Estrella CJ, García Méndez AB, Chiboub O, Hajaji S, Fernández-Astorga A, Valladares B, Martínez-Carretero E, Piñero JE, Lorenzo-Morales J (2017a) Variation in Campylobacter jejuni culturability in presence of Acanthamoeba castellanii Neff. Exp Parasitol 183(Supl):178–181. https://doi.org/10.1016/j.exppara.2017.09.005 CrossRefGoogle Scholar
- Reyes-Batlle M, Martín-Rodríguez AJ, López-Arencibia A, Sifaoui I, Liendo AR, Bethencourt Estrella CJ, García Méndez AB, Chiboub O, Hajaji S, Valladares B, Martínez-Carretero E, Piñero JE, Lorenzo-Morales J (2017b) In vitro interactions of Acanthamoeba castellanii Neff and Vibrio harveyi. Exp Parasitol 183(Supl):167–170. https://doi.org/10.1016/j.exppara.2017.09.003 CrossRefGoogle Scholar
- Schroeder JM, Booton GC, Hay J (2001) Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. J Clin Microbiol 39(5):1903–1911. https://doi.org/10.1128/JCM.39.5.1903-1911.2001 CrossRefGoogle Scholar