Test report Virkon® S - Falw.vu

The efficacy of Virkon ® S againstSalmonella paratyphi B biovar. JavaN.M.BolderAnimal Sciences Group confidential report: ID-03/0013048/BNM

The efficacy of Virkon ® S againstSalmonella paratyphi B biovar. JavaN.M.BolderAnimal Sciences Group confidential report: ID-03/0013048/BNMProject: 870-45700-3Animal Sciences Group B.V.Division Infectious DiseasesPO BOX 658200 AB LelystadThe NetherlandsN.M.BolderTel: +31 320238444e.mail: nico.bolder@wur.nlThe efficacy of Virkon ® S against Salmonella paratyphi B var. Java. N.M.Bolder, Animal Sciences Group, Lelystad, TheNetherlands. June 02, 2003.2

The efficacy of Virkon ® S against Salmonella paratyphi B biovar. JavaN.M.BolderAnimal Sciences Group, Lelystad, The NetherlandsTest report Virkon ® S, carried out according to BS.EN 1656:2000Test laboratory:Animal Sciences GroupDivision Infectious DiseasesPO BOX 65NL 8200 AB LelystadThe Netherlands.Sample:Virkon ® SBatch 4819Manufacturer:Antec International Ltd.Windham RoadChilton industrial Estate SudburySuffolk CO10 2XDUKDate of delivery: 2003-01-23Storage conditions: Room temperature at laboratoryActive substances: Not indicatedTest method and its validationMethod Dilution NeutralizationNeutralizer: Universal Neutralizer :Sodium thiosulphate: 5 g/lPhosphate buffer: 0.25N (34g KH 2 PO 4 /l distilled water): 10 mlTween 80: 30 mlL-Histidin: 1gLecithin: 3gExperimental conditionsPeriod of analysis: 2003-05-20 to 2003-05-22Appearance of product:Original: pink crystalsDilutions: pink clear solutionsProduct test concentrations:0.5% (5g/l)1.0% (10 g/l)Test temperature:10°C ± 1°CContact time:5 min ± 10 s30 min ± 10 sInterfering substanceA: low: 3 g/l bovine albuminB: high: 10 g/l yeast extract + 10 g/l bovine albuminThe efficacy of Virkon ® S against Salmonella paratyphi B var. Java. N.M.Bolder, Animal Sciences Group, Lelystad, TheNetherlands. June 02, 2003.3

Product diluent:Standardised hard water:Distilled water: 986 ml6 ml [(19,84 g anhydrous MgCl 2 /l) + (46,24 g anhydrous CaCl 2 /l)]8 ml (35.02 g NaHCO 3 /l)Incubation temperature :37°C ± 1°CBacterial counting procedure:Spread plate technique on Brilliant Green Agar, incubation at 37°C for 24 hConfirmation: biochemically and serologically of randomly selected coloniesBacterial strain used:Salmonella paratyphi B var. Java (ID-Lelystad collection, typed by National Salmonella CentreBilthoven, the Netherlands)Test results:Verification of the methodology showed no unacceptable deviations from those indicated in the BS EN1656:2000. As is indicated in table 1, the test suspension (N) contained 2,98. 10 7 cfu/ml, whereas thebacterial suspension (Nv) we used, 1,25.10 4 cfu/ml..Results of validation of neither experimental conditions (A) , nor Dilution neutralisation (C), wereinfluenced by the type of interfering fluid; high or low. Neutralisation itself did not interfere withSalmonella during the test.In Table 2 the test results are presented of decontamination trials with two concentrations of Virkon ® S,during respectively 5 and 30 minutes. The bacterial suspension (N) contained 2,31. 10 8 cfu/ml,according to the BS EN 1656:2000. The results are given as cfu/ml (Na).From these results at low dose (0,5%) of Virkon ® S , there is practically no difference in survival ofSalmonella java between the two types of interfering fluid, high or low, when the exposure time is 5minutes. When disinfection time is extended to 30 minutes, the result at low level interfering fluid issignificantly better in comparison to high level interfering fluid conditions.A similar tendency was found between high and low levels of interfering fluid during 5 minutesexposure time with 1 % of Virkon ® S, although the presence of high level of interfering materialprovided sufficient protection to Salmonella, in order to fail the conditions of EN 1656:2000. Whendisinfection time with 1% Virkon ® S was extended to 30 minutes, no surviving Salmonella java wasfound.In conclusion, reduction in viability of Salmonella Java as indicated in table 3 shows that exposuretime of 5 minutes at 1% of Virkon ® S product with low level of interfering substance, and 30 minuteswith low level of interfering substance both 0.5% and 1% of Virkon ® S and is sufficiently effective topass the requirements of EN 1656:2000. The same conclusion can be drawn for 1% of Virkon ® Sproduct when exposure time is 30 minutes with a high level of interfering substance. At these specificconditions, Salmonella was inactivated sufficiently and therefore these conditions pass therequirements of EN 1656:2000.The efficacy of Virkon ® S against Salmonella paratyphi B var. Java. N.M.Bolder, Animal Sciences Group, Lelystad, TheNetherlands. June 02, 2003.4

Table 2.Test results of two concentrations of Virkon ® S (0.5% and 1%) at 10°C ± 1°C with dilutionneutralisation method.Test microorganismSalmonellaparatyphi Bvar. Java0.5% 1% Bacterial testsuspension5 min 1 30 min 1 5 min 1 30 min 1Low interferingsubstanceVc10 -3: 488, 27210 -4 : 15, 9Low interferingsubstanceVc10 -0 :15, 010 -1 : 2, 0Low interferingsubstance0,0Low interferingsubstance0,0Vc10 -6 : 166, 40810 -7 : 10, 25(Na= 2,50. 10 5 )(Na= 4,25)(Na= 2,31. 10 6High interferingsubstanceHigh interferingsubstanceHigh interferingsubstanceHigh interferingsubstance1,54. 10 2 8,27. 10 3 3,98. 10 4 >2,31. 10 6N.M.Bolder,Animal Sciences Group,Division Infectious DiseasesPO BOX 65,NL- 8200 AB Lelystad,The NetherlandsThe efficacy of Virkon ® S against Salmonella paratyphi B var. Java. N.M.Bolder, Animal Sciences Group, Lelystad, TheNetherlands. June 02, 2003.6