Efficacy Data

Efficacy testing of ‘Virasure’ broad spectrum disinfectant against salmonid alphavirus (SAV) was performed at three different concentrations (0.1%, 1% and 2%), both in the absence and in the presence of heavy organic loading (10g/l yeast extract and 10g/l bovine albumin).

Viral testing was performed based on the protocol specified by European Standard EN14675:2006 (Chemical disinfectants and antiseptics- Quantitative suspension test for the evaluation of virucidal activity of chemical disinfectants and antiseptics used in the veterinary field- Test methods and requirements (phase 2/step1)), with some minor modifications.

Each dilution of product was tested in triplicate. As defined in the standard, a given test was considered successful if a reduction in viral titre of >4.0 log10 (104) could be shown, accompanied by acceptable controls. A given concentration was considered to pass the test when all three replicates were successful

In the absence of organic loading (hard water diluent) all three concentrations passed.In the presence of heavy loading, 0.1% failed, but passes were achieved at 1% and 2%. Further details are given in Appendix 1.

Disinfectant Efficacy Test Report-Full Report

a) Identification of the test
Laboratory:
Veterinary Sciences Division, Stoney Road, Stormont,
Belfast, BT4 3SD, Northern Ireland
b) Identification of the sample
Name of the product ‘Virasure aquatic broad spectrum disinfectant’
Batch number: 59028
Manufacturer: Clinimax
Date of delivery: 21-9-09
Expiry date: January 2011
Storage conditions: Room temperature.
Active substance(s) & concentrations Not stated
c) Experimental conditions
Period of analysis: September/November 2009
Appearance of the product and its dilutions: Fine pink powder; clear (hard water) or brown/red (heavy loading) solutions without precipitate at test dilutions
Product test: 1/1000 w/v (0.1%), 1/100 w/v (1.0%), 1/50 (2.0%)
Test temperature: 4ºC + 1ºC
Contact times: 30 min + 30 sec
Interfering substance: 10g/l yeast extract and 10g/l bovine albumin
d) Test results:
Viral strain used: Salmonid alpha virus (SAV) SPDV, pool 10, strain F93-125
Product diluent: Hard water ± interfering substance
Stability of test mixture: No precipitation of product throughout the test
Temperature of incubation: 15ºC + 1ºC
e) Locality, date and identified signature: Belfast NI. David Graham MVB PhD MRCVS

Appendix 1

Summary of viral test results (with organic loading)

Hard water (no organic loading)

Concentration Replicate Titre** Post-test Titre** Reduction Pass
Control n/a 5.75 n/a n/a
0.1% 1 <0.5 >5.25 Pass
0.1% 2 <0.5 >5.25 Pass
0.1% 3 <0.5 >5.25 Pass
1% 1 <1.5* >4.25 Pass
1% 2 <1.5* >4.25 Pass
1% 3 <1.5* >4.25 Pass
2% 1 <1.5* >4.25 Pass
2% 2 <1.5* >4.25 Pass
2% 3 <1.5* >4.25 Pass

* Cytotoxicity present ** 50% tissue culture infectious dose (TCID50)/100μl

Hard water - with organic loading

Concentration Replicate Titre** Post-test Titre** Reduction Pass
Control n/a 5.5 n/a n/a
0.1% 1 5.375 <4.0 Fail
0.1% 2 5.875 <4.0 Fail
0.1% 3 6.0 <4.0 Pass
1% 1 0.875 4.625 Pass
1% 2 1.25 4.25 Pass
1% 3 <0.5 >5.0 Pass
2% 1 <1.5* >4.0 Pass
2% 2 <1.5* >4.0 Pass
2% 3 <1.5* >4.0 Pass

* Cytotoxicity present ** 50% tissue culture infectious dose (TCID50)/100μl

Summary

In the absence of organic loading, complete inactivation of SAV was achieved at 0.1%, 1.0% and 2.0% concentrations. Note that the apparently lower reductions in titre at 1.0% and 2.0% reflect low level cytotoxicity of the product to cell culture at these concentrations rather than any demonstrated lower efficacy at these concentrations. In the presence of organic loading, all three replicates at 0.1% failed, with no reduction in titre. At both 1.0% and 2% the product passed, with all three replicates showing at least a 4 log10 reduction in titre.

Pathogen type Disease Test method Effective dilution used Testing laboratory
Birnavirus Infectious pancreatic necrosis EN 14675-2005 1/200 @ 4oC with simulated high soiling AFBI,Veterinary Sciences Division, Belfast
Rhabdovirus Viral Haemorrhagic Septicaemia EN 14675-2005 1/200 @ 4oC with simulated high soiling AFBI,Veterinary Sciences Division, Belfast
Orthomyxovirus Infectious Salmon Anaemia EN 14675-2005 1/200 @ 4oC with simulated high soiling AFBI,Veterinary Sciences Division, Belfast
Aeromonas salmonicida Furunculosis BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Renibacterium salmoninarum Bacterial kidney disease BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Yersinia ruckeri Enteric Red Mouth BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Flavobacterium psychrophilum Rainbow Trout Fry Syndrome BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Moritella viscosa Associated with Winter lesion syndrome BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Pasteurella skyensis Pasteurellosis BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Nocardia asteroides Nocardiosis BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Mycobacterium chelonae Atypical Mycobacteriosis BS EN 1656:2000 1/100 @10oC Scientific Services, Mill Farm, Norwich
Listeria monocytogenes Listeriosis BS EN 1276:1997 1/100 @10oC Scientific Services, Mill Farm, Norwich