1. Spore sampling – indoor air quality

2. Controlling fungi in your house
Fungi need food,oxygen, and water to grow
Of these, water is the most easily controlled

3. 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

4. (Semi) destructive sampling
Exploratory excavations/holes
Moderately destructive
Valuable if hidden contamination is suspected
Swab sampling
Bulk air sampling
Example: carpet samples

5. 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

6. 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

7. 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.
Culture medium affects results
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

8. Spore plate from an Anderson sampler
Medium formulation and incubation temperature important
Can impact affect spore viability?

9. 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

10. 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.

11. 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

12. 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

13. Orellanine
Cortinarius orellanoides
Kidney failure
Transplant

14. 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

15. 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

16. Ibotenic acid – muscimol
Amanita muscaria
muscle spasms, dizziness, vomiting
deep sleep/intense dreams
Abbaye de Plaincourault, Mérigny, France

17. Psilocybin-Psilocin hallucinogenic
Similar structure to bufotenin and myristicin (nutmeg)

18. Diverse gasterointestinal toxins
Agaricus, Amanita, Boletus, Chlorophyllum, Entoloma, Hebeloma, Lactarius, Marasmius, Naematoloma, Omphalotus, Pholiota, Russula, Scleroderma, Tricholoma,
B. splendidus
B. satanus

19. Tricholoma equestre (= T. flavovirens)
Muscle wasting
fatigue, weakness and myalgia
heart muscle lesions and kidney lesions