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BS EN 16438:2014 Chemical disinfectants and antiseptics. Quantitative surface test for the evaluation of fungicidal or yeasticidal activity of chemical disinfectants and antiseptics used in the veterinary area on non-porous surfaces without mechanical act, 2014
- Contents Page
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Requirements
- 5 Test method [Go to Page]
- 5.1 Principle
- 5.2 Materials and reagents [Go to Page]
- 5.2.1 Test organism
- 5.2.2 Culture media and reagents [Go to Page]
- 5.2.2.1 General
- 5.2.2.2 Water
- 5.2.2.3 Malt extract agar (MEA)
- 5.2.2.4 Diluent
- 5.2.2.5 Neutraliser
- 5.2.2.6 Hard water for dilution of products
- 5.2.2.7 Interfering substance
- 5.2.3 Test surface
- 5.3 Apparatus and glassware [Go to Page]
- 5.3.1 General
- 5.3.2 Usual microbiological laboratory equipment ) [Go to Page]
- 5.3.2.1 Apparatus for sterilisation (moist and dry heat)
- 5.3.2.2 Water bath, capable of being controlled at 10 °C ± 1 °C, 20 °C ± 1 °C, and 45 °C ± 1 °C (to maintain melted MEA. If pour plate technique is used), and at additional temperatures ± 1 °C (5.5.1).
- 5.3.2.3 Incubator, capable of being controlled at 30 °C ± 1 °C and 37 °C ± 1 °C (for drying surfaces)
- 5.3.2.4 pH-metre, having an accuracy of calibration of 0,1 pH units at 20 °C ± 1 °C.
- 5.3.2.5 Stopwatch
- 5.3.2.6 Shakers
- 5.3.2.7 Membrane filtration apparatus, constructed of a material compatible with the substances to be filtered with a filter holder of at least 50 ml volume and suitable for use with filters of diameter 47 mm to 50 mm and 0,45 µm pore size for filtration of hard water (5.2.2.6) and bovine albumin (5.2.2.7.2, 5.2.2.7.3).
- 5.3.2.8 Refrigerator capable of being controlled at 2 °C to 8 °C.
- 5.3.2.9 Graduated pipettes, of nominal capacities 10 ml and 1 ml and 0,1 ml or calibrated automatic pipettes.
- 5.3.2.10 Petri dishes, (plates) of size 90 mm to 100 mm.
- 5.3.2.11 Glass beads, diameter: 3 mm to 4 mm.
- 5.3.2.12 Volumetric flasks
- 5.3.2.13 Temperature controlled cabinet, capable of being controlled at 10 °C ± 1 °C.
- 5.3.2.14 Glass screw top container, with a base diameter of 4 cm - 5 cm.
- 5.3.2.15 Fritted filter, with porosity of 40 µm to 100 µm according to ISO 4793.
- 5.3.2.16 Flasks with vented caps.
- 5.3.2.17 Microscope capable of x 400 magnification.
- 5.3.2.18 Vacuum desicator capable of achieving a vacuum of 20-25 in. mercury.
- 5.4 Preparation of test organism suspensions and product test solutions [Go to Page]
- 5.4.1 Test organism suspension (test and validation suspension) [Go to Page]
- 5.4.1.1 General
- 5.4.1.2 Preservation and stock cultures of test organisms
- 5.4.1.3 Working culture of test organisms
- 5.4.1.4 Test suspension (N)
- 5.4.1.5 Incubation and counting of the test suspension
- 5.4.2 Product test solutions
- 5.5 Procedure for assessing the fungicidal or yeasticidal activity of the product [Go to Page]
- 5.5.1 General [Go to Page]
- 5.5.1.1 Experimental conditions (obligatory and additional)
- 5.5.1.2 Neutralisation
- 5.5.1.3 General instructions for validation and control procedures
- 5.5.1.4 Equilibration of temperature
- 5.5.2 Test procedure (Dilution-neutralisation method) [Go to Page]
- 5.5.2.1 General
- 5.5.2.2 Test “Na” - determination of fungicidal or yeasticidal concentrations
- 5.5.2.3 Water control “Nw”
- 5.5.2.4 Neutraliser control “B” - verification of the absence of toxicity of the neutraliser
- 5.5.2.5 Method validation C - dilution-neutralisation validation
- 5.5.2.6 Incubation and counting of the test, control and validation neutralisers
- 5.5.3 Observation of the test surface agar
- 5.6 Experimental data and calculation [Go to Page]
- 5.6.1 Explanation of terms and abbreviations [Go to Page]
- 5.6.1.1 Overview of the different suspensions and test mixtures
- 5.6.1.2 Vc values
- 5.6.2 Calculation [Go to Page]
- 5.6.2.1 General
- 5.6.2.2 Determination of Vc values
- 5.6.2.3 Calculation of N
- 5.6.2.4 Calculation of Na and Nw
- 5.6.2.5 Calculation of B and C
- 5.7 Verification of methodology [Go to Page]
- 5.7.1 General
- 5.7.2 Control of weighted mean counts
- 5.7.3 Basic limits
- 5.7.4 Microscopic observation
- 5.8 Expression of results and precision [Go to Page]
- 5.8.1 Reduction
- 5.8.2 Control of active and non-active product test solution (5.4.2)
- 5.8.3 Fungicidal or yeasticidal concentration [Go to Page]
- 5.8.3.1 Fungicidal concentration
- 5.8.3.2 Yeasticidal concentration
- 5.8.4 Precision, repetitions
- 5.9 Interpretation of results - conclusion [Go to Page]
- 5.9.1 General
- 5.9.2 Fungicidal or yeasticidal activity for general purposes
- 5.9.3 Qualification for certain fields of application
- 5.10 Test report
- Annex A (informative)Referenced strains of national collections
- Annex B (informative)Examples of neutralisers of the residual antimicrobial activity of chemical disinfectants and antiseptics
- Annex C (informative)Graphical representations of dilution-neutralisation method
- Annex D (informative)Example of a typical test report [Go to Page]
