Rapid efficacy testing of cattle footbath disinfectants by flow cytometry: a laboratory-validated screening method complementary to EN 1656
References
- Ariza J.M., Relun A., Bareille N., Oberle K., Guatteo R.: Effectiveness of collective treatments in the prevention and treatment of bovine digital dermatitis lesions: A systematic review. J Dairy Sci 2017, 100, 7401–7418, doi: 10.3168/jds.2016-11875.
- Berney M., Hammes F., Bosshard F., Weilenmann H.U., Egli T.: Assessment and interpretation of bacterial viability by using the LIVE/DEAD BacLight Kit in combination with flow cytometry. Appl Environ Microbiol 2007, 73, 3283–3290, doi: 10.1128/AEM.02750-06.
- Boulos L., Prévost M., Barbeau B., Coallier J., Desjardins R.: LIVE/DEAD BacLight: application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water. J Microbiol Methods 1999, 37, 77–86, doi: 10.1016/s0167-7012(99)00048-2.
- Brown M.R., Gilbert P.: Sensitivity of biofilms to antimicrobial agents. J Appl Bacteriol 1993, 74 Suppl, 87S–97S, doi: 10.1111/j.1365-2672.1993.tb04345.x.
- Claveau L., Hudson N., Jarvis P., Jeffrey P., Hassard, F.: Microbial water quality investigation through flow cytometry fingerprinting: from source to tap. Sustain Microbiol 2024, 1, qvae003, doi: 10.1093/sumbio/qvae003.
- Cook N.B., Rieman J., Gomez A., Burgi K.: Observations on the design and use of footbaths for the control of infectious hoof disease in dairy cattle. Vet J 2012, 193, 669–673, doi: 10.1016/j.tvjl.2012.06.051.
- Ersoy Z.G., Dinc O., Cinar B., Gedik S.T., Dimoglo A.: Comparative evaluation of disinfection mechanism of sodium hypochlorite, chlorine dioxide and electroactivated water on Enterococcus faecalis. LWT 2019, 102, 205–213, doi: 10.1016/j.lwt.2018.12.041.
- European Committee for Standardization (CEN): European Standard EN 1656:2020 Chemical disinfectants and antiseptics – Quantitative suspension test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in the veterinary area – Test methods and requirements (phase 2, step 1). CEN, Brussels, Belgium, 2020.
- Gerba C.P.: Quaternary ammonium biocides: efficacy in application. Appl Environ Microbiol 2015, 81, 464–469, doi: 10.1128/AEM.02633-14.
- Grootaert C., Boon N., Zeka F., Vanhoecke B., Bracke M., Verstraete W., Van de Wiele T.: Adherence and viability of intestinal bacteria to differentiated Caco-2 cells quantified by flow cytometry. J Microbiol Methods 2011, 86, 33–41, doi: 10.1016/j.mimet.2011.03.011.
- Johnston J.L., Billington S.J., Haring V., Rood J.I.: Complementation analysis of the Dichelobacter nodosus fimN, fimO, and fimP genes in Pseudomonas aeruginosa and transcriptional analysis of the fimNOP gene region. Infect Immun 1998, 66, 297–304, doi: 10.1128/IAI.66.1.297-304.1998.
- Lambert P.A.: Cellular impermeability and uptake of biocides and antibiotics in gram-positive bacteria and mycobacteria. J Appl Microbiol 2002, 92, 46S–54S, doi: 10.1046/j.1365-2672.92.5s1.7.x.
- Lindivat M., Bratbak G., Larsen A., Hess-Erga O.K., Hoell I.A.: Flow cytometric analysis of bacterial protein synthesis: monitoring vitality after water treatment. Front Microbiol 2021, 12, 772651, doi: 10.3389/fmicb.2021.772651.
- Maillard J.Y., Pascoe M.: Disinfectants and antiseptics: mechanisms of action and resistance. Nat Rev Microbiol 2024, 22, 4–17, doi: 10.1038/s41579-023-00958-3.
- Massicotte R., Mafu A.A., Ahmad D., Deshaies F., Pichette G., Belhumeur P.: Comparison between flow cytometry and traditional culture methods for efficacy assessment of six disinfectant agents against nosocomial bacterial species. Front Microbiol 2017, 8, 112, doi: 10.3389/fmicb.2017.00112.
- McDonnell G.E.: Antisepsis, disinfection, and sterilization: Types, action, and resistance. John Wiley & Sons, Hoboken, NJ, USA, 2020.
- McFeters G.A., Egli T., Wilberg E., Alder A., Schneider R., Suozzi M., Giger W.: Activity and adaptation of nitrilotriacetate (NTA)-degrading bacteria: field and laboratory studies. Water Res 1990, 24, 875–881, doi: 10.1016/0043-1354(90)90137-u.
- O’Hara C.M., Brenner F.W., Steigerwalt A.G., Hill B.C., Holmes B., Grimont P.A., Hawkey P.M., Penner J.L., Miller J.M., Brenner D.J.: Classification of Proteus vulgaris biogroup 3 with recognition of Proteus hauseri sp. nov., nom. rev. and unnamed Proteus genomospecies 4, 5 and 6. Int J Syst Evol Microbiol 2000, 50, 1869–1875, doi: 10.1099/00207713-50-5-1869.
- Ölmez H., Kretzschmar U.: Potential alternative disinfection methods for organic fresh-cut industry for minimising water consumption and environmental impact. LWT 2009, 42, 686–693, doi: 10.1016/j.lwt.2008.08.001.
- Park S.H., Choi M.R., Park J.W., Park K.H., Chung M.S., Ryu S., Kang D.H.: Use of organic acids to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh apples and lettuce. J Food Sci 2011, 76, M293–M298, doi: 10.1111/j.1750-3841.2011.02205.x.
- Park S.Y., Kim C.G.: A comparative study of three different viability tests for chemically or thermally inactivated Escherichia coli. Environ Eng Res 2018, 23, 282–287, doi: 10.4491/eer.2017.223.
- Pazos-Rojas L.A., Cuellar-Sánchez A., Romero-Cerón A.L., Rivera-Urbalejo A., Van Dillewijn P., Luna-Vital D.A., Muñoz-Rojas J., Morales-García Y.E.: The viable but non-culturable (VBNC) state, a poorly explored aspect of beneficial bacteria. Microorganisms 2023, 12, 39, doi: 10.3390/microorganisms12010039.
- Pîndaru A.M., Măruţescu L., Popa M., Chifiriuc M.C.: A label-free optical flow cytometry based-method for rapid assay of disinfectants’ bactericidal activity. Int J Mol Sci 2024, 25, 7158, doi: 10.3390/ijms25137158.
- Rosario-Cruz Z., Eletsky A., Daigham N.S., Al-Tameemi H., Swapna G.V.T., Kahn P.C., Szyperski T., Montelione G.T., Boyd J.M.: The copBL operon protects Staphylococcus aureus from copper toxicity: CopL is an extracellular membrane-associated copper-binding protein. J Biol Chem 2019, 294, 4027–4044, doi: 10.1074/jbc.RA118.004723.
- Seo Y.S., Lee G., Song S., Kim K., Cho M.: Combinatorial treatment using citric acid, malic acid, and phytic acid for synergistic inactivation of foodborne pathogenic bacteria. Korean J Chem Eng 2021, 38, 826–832, doi: 10.1007/s11814-021-0751-2.
- Solano L., Barkema H.W., Pickel C., Orsel K.: Effectiveness of a standardised footbath protocol for prevention of digital dermatitis. J Dairy Sci 2017, 100, 1295–1307, doi: 10.3168/jds.2016-11464.
- Wang Y., Hammes F., De Roy K., Verstraete W., Boon N.: Past, present and future applications of flow cytometry in aquatic microbiology. Trends Biotechnol 2010, 28, 416–424, doi: 10.1016/j.tibtech.2010.04.006.
- Wilks S.A., Michels H., Keevil C.W.: The survival of Escherichia coli O157 on a range of metal surfaces. Int J Food Microbiol 2005, 105, 445–454, doi: 10.1016/j.ijfoodmicro.2005.04.021.
- Ye C., Chen C., Feng M., Ou R., Yu X.: Emerging contaminants in the water environment: Disinfection-induced viable but non-culturable waterborne pathogens. J Hazard Mater 2024, 461, 132666, doi: 10.1016/j.jhazmat.2023.132666.
- Zdeb M., Zamorska J., Domoń A.: Disinfection of rainwater as a way to their microbiological stability and safe use. E3S Web Conf 2018, 44, 1–7, doi: 10.1051/e3sconf/20184400199.
DOI: https://doi.org/10.2478/jvetres-2026-0038 | Journal eISSN: 2450-8608 (formerly 2300-3235)
Language: English
Submitted on: Jan 14, 2026
Accepted on: Jul 13, 2026
Published on: Jul 18, 2026
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Related subjects:
© 2026 Lucie Langová, Petr Kouřil, Vladimír Zmrhal, Lenka Hromádková, Miroslav Macháček, Libor Kalhotka, Zdeněk Havlíček, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.