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Cited by 19Legionella and Acanthamoeba researchAntimicrobial agents and applicationsWater Treatment and Disinfection

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Use of 5-Cyano-2,3-Ditolyl-Tetrazolium Chloride Staining as an Indicator of Biocidal Activity in a Rapid Assay for Anti-Acanthamoeba Agents

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Kobayashi, Takeshi, Mito, Tsuyoshi, Watanabe, Narumi, Suzuki, Takashi, Shiraishi, Atsushi, Ohashi, Yuichi·2012·Journal of Clinical Microbiology
DOI 10.1128/jcm.06461-11PMC PMC3347101
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Abstract

The usefulness of 5-cyano-2,3-ditolyl-tetrazolium chloride (CTC) staining to determine the respiratory activity of Acanthamoeba was evaluated in this study. Acanthamoeba trophozoites and cysts have a red fluorescence after staining with CTC. To determine the effectiveness of CTC staining as a CTC biocidal assay for Acanthamoeba , the trophozoites and cysts of Acanthamoeba castellanii (ATCC 5037) were treated with serial concentrations of disinfectant solutions, namely, polyhexamethylene biguanide (PHMB) and commercial soft contact lens (SCL) disinfectant solutions. The treated Acanthamoeba organisms were stained with CTC, and their respiratory activity was determined by the intensity of fluorescence in a fluorescence microplate reader. The survival rates of the same samples were determined by a culture-dependent biocidal assay using the Spearman-Karber method. Our results showed that the respiratory activities determined by the CTC biocidal assay and the survival rates determined by the culture-dependent biocidal assay for Acanthamoeba trophozoites and cysts decreased in a dose-dependent way after PHMB treatments, and the results were significantly correlated ( r = 0.83 and P < 0.01 for trophozoites; r = 0.60 and P < 0.01 for cysts; Spearman rank correlation test). The respiratory activities in the trophozoites and cysts treated with SCL disinfectant solutions were significantly correlated with the survival rate ( r = 0.70 and P < 0.01 for trophozoites; r = 0.64 and P < 0.01 for cysts; Spearman rank correlation test). The significant correlation of the results indicated that the CTC biocidal assay can be used as an alternative method to a culture-dependent biocidal assay. The CTC biocidal assay is a rapid and simple method to test the effectiveness of disinfectant solutions against Acanthamoeba trophozoites and cysts.

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References 32 · Cited by 19

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Metadata

Authors
Kobayashi, T., Mito, T., Watanabe, N., Suzuki, T., Shiraishi, A., & Ohashi, Y.
Affiliations
Ehime University (JP)
ORCID
Venue
Journal of Clinical Microbiology
Publisher
American Society for Microbiology
Published
2012-05
Volume
50
Issue
5
Pages
1606-1612
ISSN
  • 0095-1137 (ISSN-L)
  • 1098-660X
DOI
10.1128/jcm.06461-11
PMC
PMC3347101
OpenAlex
W2098239999
Cited by
19
References
32
Citations by year
2026: 1 · 2024: 2 · 2023: 4 · 2022: 1 · 2020: 1 · 2018: 2 · 2017: 2 · 2016: 1 · 2015: 2 · 2014: 1 · 2013: 1 · 2012: 1
FWCI
1.02
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Journal of Clinical Microbiology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
Japan Society for the Promotion of Science (22791665)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Legionella and Acanthamoeba research
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Antimicrobial agents and applications
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Water Treatment and Disinfection
Keywords
Acanthamoeba, Acanthamoeba castellanii, biocidal activity, disinfectant efficacy, polyhexamethylene biguanide, trophozoites, cysts, respiratory activity, fluorescent staining
MeSH terms
Biguanides, Disinfectants, Fluorometry, Humans, Staining and Labeling, Tetrazolium Salts, Parasitic Sensitivity Tests, Acanthamoeba castellanii, Microbial Viability

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CoChat. (2026). Citation check: Use of 5-Cyano-2,3-Ditolyl-Tetrazolium Chloride Staining as an Indicator of Biocidal Activity in a Rapid Assay for Anti-Acanthamoeba Agents. https://app.cochat.ai/papers/10.1128/jcm.06461-11
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Kobayashi, T., Mito, T., Watanabe, N., Suzuki, T., Shiraishi, A., & Ohashi, Y. (2012). Use of 5-Cyano-2,3-Ditolyl-Tetrazolium Chloride Staining as an Indicator of Biocidal Activity in a Rapid Assay for Anti-Acanthamoeba Agents. Journal of Clinical Microbiology. https://doi.org/10.1128/jcm.06461-11