This WPG deals with the survival of avian influenza viruses in all kinds of airs and surfaces, clean or soiled.

Workpackage 5: Impact of air conditions on virus survival

WP leader: Ana Maria BURGUIERE, Institut Pasteur, France)
Participants: IPP, IPS
This WP will focus on virus survival at various hygrometry and temperature conditions. It is divided into two main parts. The first one corresponds to the set up of a system where an aerosol is generated and submitted to controlled conditions such as temperature and hygrometry (Air Experimental System). We will use an aerosol generator already available in IPP with the help of scientists with expertise in this very domain. A biocollector will be implemented inside the AES to allow for the collection of virus particles contained in the aerosols. The design of the AES including the biocollector will be done in collaboration with IPP craftsmen as well as specialist of biosafety. Once the AES is set up, investigations of the effects of various air conditions will be undertaken.
The second part of this WP is focused on surfaces exposed to the air and in particular on technical challenges to recover viruses from experimentally infected surfaces. To date there is standardised methods for the recovery of virus s from surfaces either smooth,rough or porous from the air. A peculiar study will be developed about mask surfaces and virus survival. Virus survival on fabrics and other materials used in agricultural and industrial settings or in commercial planes will also be investigated.

Workpackage 6 : Persistence of viable virus in farms surroundings

WP leader: Angel GALABOV, Institut Angelov, Sofia, Bulgaria)
Participants: MICB, IC, IPS, WIV, IPC, CIRAD, IPP
The presence of AIV A(H5N1) will be investigated in farms and in natural settings where this virus has been circulating. This will be performed in Europe (Bulgaria, Romania, France), in Africa and in Asia (Cambodia and China). In particular, litter straw, drinking and feeding bowls as well as other food and litter supplies will be screened.
Earth and passive vector can constitute surfaces where the virus can remain infectious for hours and possibly days. Earth and mechanical vectors might play a significant role in virus introduction in family or industrial farms. Moreover, available data relate to the contamination of surfaces with human strains of IVs and their results cannot be extrapolated to H5N1 AIVs without checking. Traditional and molecular techniques will be ‘imported’ from WP0, WP1 and WP5 in order to evaluate the viability of detected viruses in theses circumstances.
The following settings will be investigated:
1/ Earth and muds in and around poultry farms where there is / was an outbreak of HPAI H5N1 (special attention will be paid to the pathways used by animals between contaminated pond and barns)
2/ Earth and muds in and around areas where there is / was an outbreak of HPAI H5N1 in wild birds associated with mortality
3/ Feeding bowls, water supplies to the animals in poultry farms and other commodities where there is/was an outbreak.
Samples will be collected where farm equipments and soils are heavily dirtied by birds faeces. Faeces in general will be collected on various items, if possible, throughout the year but at least just after the outbreaks and farm cleansing. If relevant, protocols developed in WP8 for solid matrices will be implemented. If relevant and possible, earth and mud sampling will be envisaged in additional areas of bird concentration that will be followed in bird ecology studies in liaison with other tasks of this call (tasks 3 and 4 mainly).
As for it counterpart WP2, this WP was build taking into account the geographic complementarities of its participants. As for WP2, three regions of the world will be concerned by this WP: Africa, (CIRAD), Asia (IPC, IPS, WIV) and Europe (MICB, IC, IPP).

WP leader: Vincent DEUBEL, IP, China)
Participants: IPS, IPP
This WP is the counterpart of WP4 and focuses on surfaces exposed to the air and their disinfection. In the literature, it has been shown that IVs can survive for at least one day on stainless steel surfaces and for some hours on cloth, paper and tissues (Bean et al., 1982). Strangely, virus survived longer on non porous surfaces. This was probably linked to difficulties to recover virus from rough surfaces and this very issues will be dealt by WP5.
Chemical and physical conditions in various combinations will be applied to various concentrations of virus on lifeless surfaces: temperature, ultra-violet light etc …. Physical treatments in combination between them will also be experimented to evaluate their impact on virus viability and concentration: temperature, hygrometry, ultra-violet light for example.
As part of control efforts, surface treatment including the use of disinfectants will probably be needed at some stage. Regarding the disinfectants, every compound has its limitations, which are often linked to the cleanliness of the surfaces: the amount of proteins fro example. A number of disinfectants will first be selected and will include IATA approved products as well as environment friendly compounds. Virus survival will be assessed using the outcome from WP0, WP1 and WP5 and experiments will be designed according to the existing National, European or International Standards. However, regarding the virucidal activity of antiseptics and disinfectants on surfaces, there is no standard. However, it is possible to base our approach on existing ISO, European (EN) or National (eg AFNOR) standardised methods for the choice of standardised surfaces and protocols which are applied for the bactericidal and fungicidal activity of antiseptics and disinfectants such as NF T72-190 (08/1988) for steel, glass and plastic, NF EN 14349 (06/2005) for non porous surface (standardised steel) in the veterinary field. The European Standard EN 13610 (07/2003) specifies a testing method relating to the minimal virucidal activity with respect to the bacteriophages of disinfecting chemicals. EN 13610 is used in the fields of the food industry. The European Standard EN 14476 (08/2005) specifies a testing method relating to the virucidal activity of antiseptics and disinfecting chemicals for instrument surfaces and hands. A much older National Standard NF T72-180 (12/1989) relates to the virucidal activity of antiseptics and disinfectants on virus infecting Vertebrates.
This WP will primarily be done at IPS, which has the leadership. To guaranty coherence throughout the project concerning standardized assays and compliance with existing standards, IPP (Ana Maria BURGUIÈRE) will be involved in study design with the leader team.

Workpackage 8 : Evaluation of the impact of selected parameters involved in food processing

WP leader: Jean-Marie DELATTRE, Institut Pasteur de Lille, France
Animal materials involved in this project include birds faeces, blood, carcasses and food products. Naturally dead animals or experimentally infected animals or products will be considered. Methods for extraction, detection and quantification of AIV in these materials will be evaluated. With a suitable method, quantitative studies of the impact of food processing conditions on AIV will be possible. Temperature, pH, and salinity are key factors in controlling micro-organisms in food products. Thermal inactivation of viruses has proven an efficient decontamination method. Chicken is generally well cooked as are ‘white’ meats, however in some countries like France duck breast filet (magret de canard) is often served medium rare and the temperature at the centre of the piece of meat might not be sufficient for a long enough time. In addition in other part of the world, consumption of raw duck blood is not impossible and can be the source of infection. Controlling micro-organisms in food products is an increasing field of investigation, and other factors (such as pH and salt content) may positively interact with heat. It is thus essential to evaluate the effect of these factors on AIV control.
In the field of food microbiology, there is a great number of standards (ISO, European standards (EN) or National (eg AFNOR)). Some of these standards stipulate general rules for food microbiological examination (NF ISO 7218 and NF ISO 7218/A1 - December 2001), others define the methodology for samples preparation (NF ISO 6887-1 of September 1999), some of them being very specific for meat and meat derived products (eg NF ISO 6887-2 of January 2004). Nevertheless these standards relate to the analysis of the bacteriological quality of food and the detection of a limited amount of given pathogens. Since June 2005, Standard NF IN ISO 22174 outlines general requirements and establishes definitions for the detection and identification of micro-organisms (either pathogens or not) in food by Polymerisation Chain Reaction (PCR). Two draft standards PR NF ISO 20837et PR NF ISO 20838-September 2004 respectively define requirements relating to sample preparation and amplification for qualitative PCR detection of pathogens in food. Our methodological approaches should be compatible with these existing standards for the detection and quantification of AIVs in poultry meat. Institut Pasteur de Lille, which has long been specialised in food microbiology and has been involved in the making of numerous standards, has been working on the evaluation and modelling of thermal inactivation of enteric viruses in fruit as influenced by pH and sugar, or the development of detection and quantification methods of micro-organisms in food products using various methods (titration, molecular biology).