Transcript Slide 1

Spore sampling –
indoor air quality
Controlling fungi in your house


Fungi need
food,oxygen, and
water to grow
Of these, water
is the most easily
controlled
Nondestructive sampling

Tape lift samples
Most appropriate for identifying fungi causing
visible contamination
 Analyze by direct microscopy
 Identification seldom beyond genus level by spore
morphology;
 Semiquantitative

(Semi) destructive sampling

Exploratory excavations/holes
Moderately destructive
 Valuable if hidden contamination is suspected

Swab sampling
 Bulk air sampling


Example: carpet samples
Bulk air samples – non-destructive

Bulk dust – excellent natural filter/accumulator

Collect a volume of dust – good for porous
surfaces (fabric, insulation)
Can be semi-quantitative
 Sample size is important

Little equipment needed – HEPA filtered vacuum
 Comparison of air quality in different areas

+/ – indoor, outdoor
 Also samples pollens, fibreglass, particulates


Analyze by direct microscopy or culture methods
Air samplers – impact spore traps







Rapid turnaround time
Particulate matter may
interfere with analysis
Slide preparation requires
skill
Morphological similarity
some genera
Under-representation of
some genera (e. g. Stachybotrys)
Standardized reporting
Particle bounce may produce
low counts
Burkard
Rotorod
Sieve-type impactors

Anderson sampler





Culture medium affects results





1-6 stages,
multiple stage provides differentiation of
particle size
air is drawn through a sieve plate onto a
culture plate.
The plate is incubated in a laboratory, and
colonies that develop are speciated and
enumerated.
Malt Extract Agar (MEA) General screening
media for fungi, particularly hydrophilic
fungi
Dichloran Glycerol Agar (DG18)
xerotolerant fungi
Czapek Cellulose Agar (CCA) Stachybotrys
and cellulolytic Penicillium
Corn Meal Agar Stachybotrys, Cheatomium and
other hydrophilic fungi
Spore impact may reduce viability
Spore plate from an Anderson sampler


Medium formulation and incubation
temperature important
Can impact affect spore viability?
http://helios.bto.ed.ac.uk/bto/microbes/airborne.htm
Mushroom toxins

Amanitins – Amanita species,
especially phalloides and virosa


1998 A. phalloides found on Vancouver
Island
Test for amanitoxins (not for edibility)




Mushroom ‘juice’ on newspaper
dry
Add a drop of concentrated HCl
Blue reaction within 20 minutes
(amanitin+ lignin + acid) indicates
amatoxins.
Amanita virosa; A phalloides
Gallerina autumnalis; Conocybe filaris
Bastien Treatment for amanitin poisoning









A French physician, Dr. P. Bastien, has developed a new treatment for Amanita
poisoning.
(1) intravenous injections of 1 gram vitamin C twice a day
(2) two capsules of nifuroxazide three times a day
(3) two tablets of dihydrostreptomycin three times a day.
Supplemented by measures to control fluid and electrolyte balance, and by
penicillin.
Successfully treated 15 cases of A. phalloides poisoning between 1957 and 1969.
In 1974 he ate 65 grams of A. phalloides and survived.
In 1981 he ate 70 grams of A. phalloides and again successfully treated himself.
Reported that the Bastien treatment is now used throughout France, where it
saves the lives of all those whose treatment has not been delayed until massive
liver and kidney damage has occurred. This method should obviously be
widely publicized and tested in North America.
Four classes of mushroom toxins









cause extensive cell destruction,
overt symptoms only after a significant, and potentially fatal, delay
amanitin, orellanine, monomethylhydrazine, the undetermined toxin(s)
in Tricholoma equestre/T. flavovirens
autonomic nervous system
symptoms either as soon as they have been absorbed, or whenever the
appropriate substrate enters the system (muscarine, coprine)
central nervous system
symptoms as soon as they have been absorbed (muscimol, psilocybin)
Undetermined toxins that act on the alimentary canal
symptoms as soon as they have been absorbed
Mono-methylhydrazine





boiling point of 87.5C
vapours are toxic
2-4% of all fatal mushroom
poisonings
Symptoms appear 2-12 hours
(typically 6-8 hours)
Bloating, nausea, vomiting,
diarrhea and abdominal cramps,
loss of muscular control,
jaundice; convulsions, death
Orellanine



Cortinarius orellanoides
Kidney failure
Transplant
Coprine (Antabuse-like)

Acetaldehyde dehydrogenase inhibitor
hot flushes of the face and neck
 metallic taste
 tingling /numbness
 throbbing headache
 nausea and vomiting.
 possible cardiac arrhythmia

Muscarine



muscarinic adrenergic
receptor inhibitor
pupil constriction,
blurred vision, muscle
spasms, diarrhea, slow
heartbeat, and a drop
in blood pressure
PSL syndrome
perspiration, salivation,
lachrymation
Inocybe lanuginella Inocybe fastigiata
Clitocybe dealbata Inocybe geophylla
Ibotenic acid – muscimol



Amanita muscaria
muscle spasms,
dizziness, vomiting
deep sleep/intense
dreams
Abbaye de Plaincourault, Mérigny, France
Psilocybin-Psilocin


hallucinogenic
Similar structure to
bufotenin and
myristicin (nutmeg)
Diverse gasterointestinal toxins

Agaricus, Amanita, Boletus,
Chlorophyllum, Entoloma,
Hebeloma, Lactarius,
Marasmius, Naematoloma,
Omphalotus, Pholiota,
Russula, Scleroderma,
Tricholoma,
B. splendidus
B. satanus
Tricholoma equestre (= T. flavovirens)



Muscle wasting
fatigue, weakness and
myalgia
heart muscle lesions and
kidney lesions