Challenges to get axenic cultures of Trichomonas spp.: A new approach in eradication of contaminants and maintenance of laboratory microbiological cultures
2015 (English)In: Journal of Microbiological Methods, ISSN 0167-7012, E-ISSN 1872-8359, Vol. 118, 25-30 p.Article in journal (Refereed) PublishedText
Contamination of microbiological and cell cultures is a major problem in many scientific and clinical laboratories as well as bioproduct manufacturers worldwide. In the current study we established a rapid (9 day) method to detect and eliminate fungal and bacterial contamination in cultures of the unicellular eukaryote Trichomonas spp. The developed method combines identification of the contaminating microorganisms using PCR and sequencing of the 16/18S regions followed by phylogenetic analysis. The next step was a phylogeny-guided selection of antibiotic treatments. We then used a two-step propidium iodide-resorufin assay to test the effect of selected antibiotics. The result was a quick and worthwhile purification of trichomonad laboratory cultures. Our workflow may also be implemented to obtain new isolates of trichomonads from clinical samples if initial broad-spectrum antibiotic therapy fails.
Place, publisher, year, edition, pages
2015. Vol. 118, 25-30 p.
Trichomonas spp., Contamination, Resorufin, Propidium iodide, Antibiotic, Phylogeny
IdentifiersURN: urn:nbn:se:uu:diva-270634DOI: 10.1016/j.mimet.2015.08.009ISI: 000364892300004PubMedID: 26284963OAI: oai:DiVA.org:uu-270634DiVA: diva2:890223
FunderCarl Tryggers foundation , CTS KF15:6Swedish Research Council Formas, 2008-1366