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 |
| f) 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)
Titre** |
||||
Concentration |
Replicate |
Post-test |
reduction |
Pass |
Control |
n/a |
5.75 |
n/a |
n/a |
0.1% 0.1% 0.1% |
1 2 3 |
<0.5 <0.5 <0.5 |
>5.25 >5.25 >5.25 |
PASS PASS PASS |
1% 1% 1% |
1 2 3 |
<1.5* <1.5* <1.5* |
>4.25 >4.25 >4.25 |
PASS PASS PASS |
2% 2% 2% |
1 2 3 |
<1.5* <1.5* <1.5* |
>4.25 >4.25 >4.25 |
PASS PASS PASS |
**50% tissue culture infectious dose (TCID50)/100μl
n/a not applicable
Hard water-with organic loading
Titre** |
||||
Concentration |
Replicate |
Post-test |
reduction |
Pass |
Control |
n/a |
5.5 |
n/a |
n/a |
0.1% 0.1% 0.1% |
1 2 3 |
5.375 5.875 6.0 |
<4.0 <4.0 <4.0 |
FAIL FAIL FAIL |
1% 1% 1% |
1 2 3 |
0.875 1.25 <0.5 |
4.625 4.25 >5.0 |
PASS PASS PASS |
2% 2% 2% |
1 2 3 |
<1.5* <1.5* <1.5* |
>4.0 >4.0 >4.0 |
PASS PASS PASS |
**50% tissue culture infectious dose (TCID50)/100μl
n/a not applicable
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. Note though that at 1.0%, low levels of virus were still present in two of three replicates.
| 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 |
