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. 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

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