1. Measurements of biological contaminants - WP3.3 SINPHONIE Project kick-off meeting 10-12 November, REC Conference Center, Szentendre Hungary Martin Täubel, THL

2. Lead: THL - National Institute for Health and Welfare, Finland Martin Täubel, Anne Hyvärinen, Aino Nevalainen Co-Lead: NIEH - National Institute of Environmental Health, Hungary Donat Magyar et al.

3. The overview of the presentation Objectives of this work package Tasks to be completed Critical issues

4. Background Microbial pollution is a key element of indoor pollution. (WHO guidelines for indoor air quality: dampness and mould. 2009, WHO Regional Office for Europe) - exposure to indoor microbial contaminants related to adverse health outcomes - includes hundreds of species of bacteria and fungi that grow whenever sufficient moisture is available - indoor growth; outdoor sources; person/person transfer Calltext: "... In addition to above (chemical) pollutants, allergens in dust and mould and bacteria in dust and the air will be measured."

5. Objectives i) to define a common methodology to realize measurements of biological contaminants in schools across Europe ii) to produce new exposure data on biological contaminants in schools with a great European coverage iii) to apply molecular, culture-independent approaches for detection of specific bacterial/fungal groups, in addition to general markers for microbial exposure

6. Tasks 1) Definition of harmonized methodologies standardisation in protocols, equipment, field form, sample shipment 2) Organisation of training workshops centralized training of field workers; simple sampling approach 3) Quality control/Quality assurance see 1) and 2); centralized analyses of sample materials 4) Conduction of sampling campaigns

7. Environmental sampling – main study → 120 schools/kindergardens; 3 classrooms → standardised, ’light’ sampling approach → sampling equipment provided centrally; analyses centrally oSettled dust sampling:  for determination of exposure to microbes  long-term integrated sample, representing airborne exposure  4 weeks passive sample accumulation (using SDB or EDC) oFloor/furniture dust sampling:  sampling with regular vacuum cleaners equipped with ALK filter (for allergen determination) and nylon socks (back-up for microbial determination)

8. Environmental sampling – side study → assess contribution of outdoor air to indoor microbial content → conducted in 1 center per each cluster, to cover different climatic regions (total 12 schools)

9. Environmental sampling – side study

10. → assess contribution of outdoor air to indoor microbial content → conducted in 1 center per each geographical cluster, to cover different climatic regions (total 12 schools) oActive air sampling:  4 countries, 12 schools; 3 indoor / 1 outdoor per school  2 sampling campaigns (heating vs non-heating season)  engage centers with experience and equipment  active air sampling onto filters using pumps and Button aerosol sampler (tbd)  centralised analyses of microbes

11. Tasks 1) Definition of harmonized methodologies standardisation in protocols, equipment, field form, sample shipment 2) Organisation of training workshops centralized training of field workers; simple sampling approach 3) Quality control/Quality assurance see 1) and 2); centralized analyses of sample materials 4) Conduction of sampling campaigns 5) Analyses of sample materials

12. Centralised analyses of sample materials  Specific exposure to microbial groups (THL)  settled dust from main study and active air samples from side study (indoor/outdoor comparison)  quantitative PCR for different relevant mould and bacterial groups (eg. Penicillium/Aspergillus, Cladosporuim sp., Streptomyces sp.)  General microbial exposure levels (NIEH, THL)  settled dust from main study  bacterial endotoxin (NIEH) and fungal ergosterol (THL)  Analyses of indoor relevant allgergens (UU)  vacuumed floor dust from main study (subsample)  dog & cat allergen, house dust mites (University of Uppsala=UU)

13. Critical issue – time to generate the data!  Important to measure biological exposures in parallel with chemical/physical parameters and the health studies; BUT: time needed for sample processing/sample analyses/data generation need to be considered! Sample materials from field work Data to database

14. How to deal with the issue of time  Schedule centers for passive sampling for biologicals into 3 blocks:  Block 1 (mid Oct to mid Nov): norhternmost centers, who start heating season and field work the earliest  Block 2 (mid Nov to mid Dec): remaining centers that perform field work before Christmas  Block 3 (mid Jan to mid Feb): centers who perform field work after Christmas (southernmost centers)

16. How to deal with the issue of time  Schedule centers for passive sampling for biologicals into 3 blocks:  Block 1 (mid Oct to mid Nov): norhternmost centers, who start heating season and field work the earliest  Block 2 (mid Nov to mid Dec): remaining centers that perform field work before Christmas  Block 3 (mid Jan to mid Feb): centers who perform field work after Christmas (southernmost centers)  Continuous analyses of samples as they arrive in the analyzing centers  Prioritize analyses of certain biological agents that will provide data on general biological levels until April (i.e. ergosterol, endotoxin, allergens)