Measurements of biological contaminants - WP3.3SINPHONIE Project kick-off

meeting 10-12 November, REC Conference Center,

Szentendre Hungary

Martin Täubel, THL

Lead: THL - National Institute for Health and Welfare, Finland

Martin Täubel, Anne Hyvärinen, Aino Nevalainen

Co-Lead: NIEH - National Institute of Environmental Health, HungaryDonat Magyar et al.

The overview of the presentation

• Objectives of this work package

• Tasks to be completed

• Critical issues

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."

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

Tasks

1) Definition of harmonized methodologiesstandardisation in protocols, equipment, field form, sample shipment

2) Organisation of training workshopscentralized training of field workers; simple sampling approach

3) Quality control/Quality assurancesee 1) and 2); centralized analyses of sample materials

4) Conduction of sampling campaigns

Environmental sampling – main study

→ 120 schools/kindergardens; 3 classrooms

→ standardised, ’light’ sampling approach

→ sampling equipment provided centrally; analyses centrally

o Settled dust sampling: for determination of exposure to microbes long-term integrated sample, representing airborne exposure 4 weeks passive sample accumulation (using SDB or EDC)

o Floor/furniture dust sampling: sampling with regular vacuum cleaners equipped with ALK

filter (for allergen determination) and nylon socks (back-up for microbial determination)

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)

Environmental sampling – side study

Cluster 1

Cluster 2Cluster 3

Cluster 4

Cluster 1: Northern Europe-Cold climate, cold winters-Large differences old vs new buildings ; Well insulated rooms, ventilation systems (Norway, Sweden, Finland)

Cluster 2: Central Europe-Moderate climate, moderately cold winters-Differences old vs new buildings (ventilation, insulation, passive and low energy construction)

Cluster 3: Eastern Europe-Colder climate, cold winters-Moderate to low insulation no ventilation systems (?)

Cluster 4: Southern Europe-Warm , warmer winters Mediterranean climate-Moderate to low insulation-Ventilation?

Environmental sampling – side study→ 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)

o Active 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

Tasks

1) Definition of harmonized methodologiesstandardisation in protocols, equipment, field form, sample shipment

2) Organisation of training workshopscentralized training of field workers; simple sampling approach

3) Quality control/Quality assurancesee 1) and 2); centralized analyses of sample materials

4) Conduction of sampling campaigns

5) Analyses of sample materials

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)

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

mo1 mo2 mo3 mo4 mo5 mo6

Processing of dust and splitting (N=500)

DNA extraction (N=500)

qPCR analyses + data (N=500*7)

Ergosterol analyses + data (N=400)

Endotoxin analyses + data (N=400)

Allergen analyses + data (N=200*2+120)

How to deal with the issue of time1) Schedule centers for passive sampling for biologicals into 3 blocks:

1) Block 1 (mid Oct to mid Nov): norhternmost centers, who start heating season and field work the earliest

2) Block 2 (mid Nov to mid Dec): remaining centers that perform field work before Christmas3) Block 3 (mid Jan to mid Feb): centers who perform field work after Christmas (southernmost centers)

How to deal with the issue of time1) 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)

2) Continuous analyses of samples as they arrive in the analyzing centers

3) Prioritize analyses of certain biological agents that will provide data on general biological levels until April (i.e. ergosterol, endotoxin, allergens)