Fungal DNA health studies and

patient cases

Guihong Cai

Occupational hygienist (AMM) and

Researcher (pos-doc): Department of Allergy

and Sleep Research, Uppsala University.

email: guihong.cai@medsci.uu.se

Contents

2

Fungal DNA method (QPCR)

Malaysia School Study

HESE study: Health Effects of School Environment

Investigation of patient cases with DNA method

EpiHealth study

Molds/Dampness and Health

3

Over 100 epidemiological studies

showed that dampness/moulds exposure

are at increased risk of respiratory

symptoms/infection, asthma, headache,

eye symptoms or SBS, but depending on

the species involved (Quansah, et al.

2012 ; WHO 2009; IOM 2004).

Viable/Non-viable moulds could be

risk to health, both need to accurately

measure specially the species

composition and concentrations.

4

Methods for Mould Measurements

Fig. qPCR and culture-based methods, 4 Aspergillus spp. (GM levels), home

dust samples in Cincinnati, OH. (Meklin et al., 2004)

Traditional culture/microscope counting cells method:

Economical/on species level, but requires qualified person or only

detect the viable/cultural moulds.

Quantitative Polymerase Chain Reaction (qPCR): Fast,

quantify viable/non-viable moulds, low detection limit and high

accuracy, can detect general (e.g. Asp/Pen) and specific fungal DNA

sequences (e.g. A. versicolor).

1. Cotton Swab:

60 cm2 upper doorframe

2017/2/3

2. Petri dish:

Open to indoor air

3. Vacuum cleaner:

ALK filters

Particles/Dust sampling

Malaysia School Study (I):

Population

8 schools (32 classrooms),

462 pupils participated.

Participation rate: 96 %; Mean

age: 14yr (range 14-16).

Johor Bahru

Fungal DNA Cotton swab (CE/m2) Petri dish (CE/m2/day)

Total fungal DNA

(GM±GSD1) 5.7×108±2.1 9.2×106±1.9

Asp/Pen DNA

(GM±GSD1) 0.5×108±3.3 1.6×106±3.0

A. versicolor DNA

(Arithmetic Mean) 8 780 1 478

S. chartarum DNA

(Arithmetic Mean) 26 105

Streptomyces DNA

(Arithmetic Mean) 893 1 271

2017/2/3 7

Result (I-1):

Fungal DNA Levels

1 GM: Geometric mean, GSD: Geometric standard deviation.

Result (I-2) Fungal DNA/Health Associations

Fungal DNA

(Petri dish dust)

Wheeze Daytime

breathlessness

Doctor-

diagnosed asthma

1Total fungal DNA 0.71 (0.32-1.59) 1.02 (0.66-1.58) 1.32 (0.89-1.95)

2Asp/Pen DNA 0.84 (0.21-3.35) 0.91 (0.40-2.08) 0.99 (0.40-2.44)

3A. versicolor DNA 1.19 (1.07-1.32)** 1.14 (1.02-1.28)* 1.04 (0.93-1.15)

4S. chartarum DNA 1.03 (0.90-1.18) 0.88 (0.80-0.98)* 1.01 (0.92-1.11)

5Streptomyces DNA 0.95 (0.73-1.24) 1.05 (0.89-1.24) 1.18 (1.00-1.40)*

No associations between fungal DNA in

swab samples and respiratory health.

Fungal DNA in HESE study

Result (II-1):

log-Total Fungal DNA Levels

2017/2/3 10

log

-to

tal

fu

ng

al D

NA

(C

E/g

du

st) 6.5

6.0

5.5

5.0

4.

4.0

Siena,

Italy

Udine,

Italy

Oslo,

Norway

Uppsala,

Sweden

Århus,

Denmark

Reims,

France

Simoni et al., 2011

2017/2/3 11

0

0.5

1

1.5

2

2.5

3

3.5

4 2nd tertile

3rd tertile

Wheeze Dry cough

at night Rhinitis Cough

Result (II-2) Associations by Tertile:

Total Fungal DNA/Health

Risk (Odds Ratio and 95 % Confidence Interval) for reporting respiratory disorders by tertile

exposure to log-Total funal DNA (CE/g).

Analyses accounted for gender, age , ETS, and lifetimeasthma).

Reference category for moulds exposure: 1st tertile of the distribution

Dose Effect

2017/2/3 12

0

0.5

1

1.5

2

2.5

3

3.5

4 2nd tertile

3rd tertile

Wheeze dry cough

at night Rhinitis Cough

Analyses accounted for gender, age , ETS, and lifetimeasthma).

Reference category for moulds exposure: 1st tertile of the distribution.

Risk (Odds Ratio and 95 % Confidence Interval)

for reporting respiratory disorders by tertile exposure

to log-Asp/Pen DNA (CE/g).

0

1

2

3

4

5

6 2nd tertile

3rd tertile

Wheeze Dry cough

at night Rhinitis Cough

Risk (Odds Ratio and 95 % Confidence Interval)

for reporting respiratory disorders by tertile exposure

to log-A. versicolor DNA (CE/g).

Dose Effect

Result (II-3) Associations by Tertile:

Fungal DNA/Health

2017/2/3 13

Result (II-4) Lung Function

Linear regression models

Crude adjusteda

Log-total VM

FEV1 -0.11 (-0.24;0.02)bl -0.05 (-0.07;0.18)

FVC -0.21 (-0.36;-0.06) 0.01 (-0.12 ;0.14)

Log-A. versicolor DNA

FEV1 -0.08 (-0.16;-0.01)* -0.04 (-0.14 ;0.06)

FVC -0.16 (-0.25;-0.06)** -0.10 (-0.21;0.01)bl

Log-Streptomyces DNA

FEV1 -0.13 (-0.21;-0.04)** -0.14 (-0.23 ;-0.05)*

FVC -0.15 (-0.26;-0.05)** -0.14 (-0.23 ;-0.04)**

FEV1 = forced expiratory volume in 1 second; FVC = forced vital capacity.

p<0.05, ** p<0.01, bl 0.05 < p < 0.1. aAdjusted for gender, age, height, passive smoking at home, and any of lifetime-

asthma, dry cough at night, cough.

Anozona DNA method

2017/2/3 14

1. Total fungi: common indoor mould.

2. Asp/Pen: like warm, allergic

3. A.versicolor: good indicator of mild dampness in nordic

country (no need high humidity), health problems: inflamation

in mice experiment (1×106 spores) (Jussila and Komulainen,

2002).

4. S.chartarum: well known toxic spp.

5. Streptomyces: health problems, common spp. and soil odour

Fungal DNA analysis done by anoZona AB, Uppsla

2017/2/3 15

Fungal DNA Living room (at

1st floor)

Sleeping room

(at 2nd floor)

Total fungal DNA 30 417 8 853

Asp/Pen DNA 4 015 1 052

A. versicolor DNA 22 5

S. chartarum DNA 0 0

Streptomyces DNA 11 6

Two-floors wooden house (1932) with water damage at the 2nd floor

(damaged floorboards were removed).

By visiting: we found moisture damaged signs in bathrooms at the 1st

floor

Patient case 1: home environment

2017/2/3 16

Fungal DNA Grodan section Haren section

Total fungal DNA 13 916 36 866

Asp/Pen DNA 665 942

A. versicolor DNA 2 0

S. chartarum DNA 0 0

Streptomyces DNA 5 8

Patient is experiencing the most symptoms and smell in the “Haren”

section.

Significantly less health problems and odor in “Grodan” section.

Patient case 2: day care centre

Conclusions

With DNA method: will always find indoor mold, and sometimes

found the hidden damage, but the content was higher in the room

where the patient is experiencing more symptoms.

With DNA method: can also get information about the renovation

of dampness has been successful.

The method is useful especially when there is no building

technical investigation.

When communicating with the patient: There is not a simple

relationship between the level of fungal DNA and health, and

emphasizes that this method is a way to better assess whether the

building is a damp building.

2017/2/3 17

2017/2/3 18

EpiHealth study: Primary objective was to

study how interactions between lifestyle factors

and genotypes contribute to the development of

common disorders (eg. respiratory diseases, lung

function) in humans (started in 2011) with 19 135

participants, age (45-75 ys old).

My research now: Study the body mass index

(BMI) scores, weight gain, obesity and central

obesity associated with the prevalence of sleep

disturbances and the impact of sleep duration and

insomnia symptoms when other lifestyle variables

were taking into account.

My research at Dep.

Allergy and Sleep Research

0

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