Spore sampling Spore sampling – indoor air – indoor air

qualityquality

Controlling fungi in your Controlling fungi in your househouse

Fungi need Fungi need food,oxygen, food,oxygen, and water to and water to growgrow

Of these, Of these, water is the water is the most easily most easily controlledcontrolled

Nondestructive sampling Nondestructive sampling Tape lift samples Tape lift samples

Most appropriate for identifying fungi Most appropriate for identifying fungi causing visible contaminationcausing visible contamination

Analyze by direct microscopy Analyze by direct microscopy Identification seldom beyond genus Identification seldom beyond genus

level by spore morphology;level by spore morphology; SemiquantitativeSemiquantitative

(Semi) destructive (Semi) destructive samplingsampling

Exploratory excavations/holes Exploratory excavations/holes Moderately destructiveModerately destructive Valuable if hidden contamination is Valuable if hidden contamination is

suspectedsuspected Swab sampling Swab sampling Bulk air samplingBulk air sampling

Example: carpet samplesExample: carpet samples

Bulk air samples – non-Bulk air samples – non-destructivedestructive

Bulk dust – excellent natural Bulk dust – excellent natural filter/accumulatorfilter/accumulator Collect a volume of dust – good for porous Collect a volume of dust – good for porous

surfaces (fabric, insulation)surfaces (fabric, insulation) Can be semi-quantitativeCan be semi-quantitative Sample size is importantSample size is important

Little equipment needed – HEPA filtered Little equipment needed – HEPA filtered vacuumvacuum

Comparison of air quality in different areasComparison of air quality in different areas +/ – indoor, outdoor+/ – indoor, outdoor Also samples pollens, fibreglass, particulatesAlso samples pollens, fibreglass, particulates

Analyze by direct microscopy or culture Analyze by direct microscopy or culture methodsmethods

Air samplers – impact Air samplers – impact spore trapsspore traps

Rapid turnaround timeRapid turnaround time Particulate matter may Particulate matter may

interfere with analysis interfere with analysis Slide preparation Slide preparation

requires skillrequires skill Morphological Morphological

similarity some genera similarity some genera Under-representation Under-representation

of some genera (of some genera (e. g.e. g. StachybotrysStachybotrys))

Standardized reporting Standardized reporting Particle bounce may Particle bounce may

produce low countsproduce low counts

Burkard

Rotorod

Sieve-type impactorsSieve-type impactors Anderson samplerAnderson sampler

1-6 stages, 1-6 stages, multiple stage provides multiple stage provides

differentiation of particle sizedifferentiation of particle size air is drawn through a sieve plate air is drawn through a sieve plate

onto a culture plate. onto a culture plate. The plate is incubated in a The plate is incubated in a

laboratory, and colonies that laboratory, and colonies that develop are speciated and develop are speciated and enumerated. enumerated.

Culture medium affects resultsCulture medium affects results Malt Extract Agar (MEA) General Malt Extract Agar (MEA) General

screening media for fungi, screening media for fungi, particularly hydrophilic fungiparticularly hydrophilic fungi

Dichloran Glycerol Agar (DG18) Dichloran Glycerol Agar (DG18) xerotolerant fungixerotolerant fungi

Czapek Cellulose Agar (CCA) Czapek Cellulose Agar (CCA) StachybotrysStachybotrys and cellulolytic and cellulolytic PenicilliumPenicillium

Corn Meal AgarCorn Meal Agar Stachybotrys, Stachybotrys, Cheatomium Cheatomium andand other other hydrophilic fungihydrophilic fungi

Spore impact may reduce viabilitySpore impact may reduce viability

Spore plate from an Anderson Spore plate from an Anderson samplersampler

Medium formulation and incubation Medium formulation and incubation temperature important temperature important

Can impact affect spore viability?Can impact affect spore viability?

http://helios.bto.ed.ac.uk/bto/microbes/airborne.htm

Mushroom toxinsMushroom toxins

Amanitins – Amanita species, Amanitins – Amanita species, especially phalloides and especially phalloides and virosavirosa 1998 1998 A. phalloidesA. phalloides found on found on

Vancouver IslandVancouver Island Test for amanitoxins (not for Test for amanitoxins (not for

edibility)edibility) Mushroom ‘juice’ on Mushroom ‘juice’ on

newspapernewspaper drydry Add a drop of concentrated Add a drop of concentrated

HClHCl Blue reaction within 20 Blue reaction within 20

minutes (amanitin+ lignin + minutes (amanitin+ lignin + acid) indicates amatoxins. acid) indicates amatoxins.

Amanita virosa; A phalloides

Gallerina autumnalis; Conocybe filaris

Bastien Treatment for amanitin Bastien Treatment for amanitin poisoningpoisoning

A French physician, Dr. P. Bastien, has developed a new A French physician, Dr. P. Bastien, has developed a new treatment for treatment for AmanitaAmanita poisoning. poisoning.

(1) intravenous injections of 1 gram vitamin C twice a day(1) intravenous injections of 1 gram vitamin C twice a day (2) two capsules of nifuroxazide three times a day(2) two capsules of nifuroxazide three times a day (3) two tablets of dihydrostreptomycin three times a day. (3) two tablets of dihydrostreptomycin three times a day.

Supplemented by measures to control fluid and electrolyte Supplemented by measures to control fluid and electrolyte balance, and by penicillin. balance, and by penicillin.

Successfully treated 15 cases of Successfully treated 15 cases of A. phalloidesA. phalloides poisoning poisoning between 1957 and 1969. between 1957 and 1969.

In 1974 he ate 65 grams of In 1974 he ate 65 grams of A. phalloidesA. phalloides and survived. and survived. In 1981 he ate 70 grams of In 1981 he ate 70 grams of A. phalloidesA. phalloides and again and again

successfully treated himself. successfully treated himself. Reported that the Bastien treatment is now used Reported that the Bastien treatment is now used

throughout France, where it saves the lives of all those throughout France, where it saves the lives of all those whose treatment has not been delayed until massive liver whose treatment has not been delayed until massive liver and kidney damage has occurred. This method should and kidney damage has occurred. This method should obviously be widely publicized and tested in North America. obviously be widely publicized and tested in North America.

Four classes of Four classes of mushroom toxinsmushroom toxins

cause extensive cell destruction, cause extensive cell destruction, overt symptoms only after a significant, and potentially overt symptoms only after a significant, and potentially

fatal, delay fatal, delay amanitin, orellanine, monomethylhydrazine, the amanitin, orellanine, monomethylhydrazine, the

undetermined toxin(s) in undetermined toxin(s) in Tricholoma equestre/T. Tricholoma equestre/T. flavovirensflavovirens

autonomic nervous systemautonomic nervous system symptoms either as soon as they have been absorbed, or symptoms either as soon as they have been absorbed, or

whenever the appropriate substrate enters the system whenever the appropriate substrate enters the system (muscarine, coprine)(muscarine, coprine)

central nervous systemcentral nervous system symptoms as soon as they have been absorbed (muscimol, symptoms as soon as they have been absorbed (muscimol,

psilocybin)psilocybin)

Undetermined toxins that act on the alimentary canalUndetermined toxins that act on the alimentary canal symptoms as soon as they have been absorbed symptoms as soon as they have been absorbed

Mono-methylhydrazineMono-methylhydrazine boiling point of 87.5Cboiling point of 87.5C vapours are toxic vapours are toxic 2-4% of all fatal mushroom 2-4% of all fatal mushroom

poisoningspoisonings Symptoms appear 2-12 Symptoms appear 2-12

hours (typically 6-8 hours) hours (typically 6-8 hours) Bloating, nausea, Bloating, nausea,

vomiting, diarrhea and vomiting, diarrhea and abdominal cramps, loss of abdominal cramps, loss of muscular control, muscular control, jaundice; convulsions, jaundice; convulsions, deathdeath

Orellanine Orellanine

Cortinarius orellanoidesCortinarius orellanoides Kidney failureKidney failure TransplantTransplant

Coprine (Antabuse-like)Coprine (Antabuse-like)

Acetaldehyde dehydrogenase Acetaldehyde dehydrogenase inhibitorinhibitor hot flushes of the face and neckhot flushes of the face and neck metallic tastemetallic taste tingling /numbness tingling /numbness throbbing headachethrobbing headache nausea and vomiting. nausea and vomiting. possible cardiac arrhythmiapossible cardiac arrhythmia

Muscarine Muscarine

Inocybe lanuginella Inocybe fastigiata Clitocybe dealbata Inocybe geophylla

muscarinic muscarinic adrenergic receptor adrenergic receptor inhibitorinhibitor

pupil constriction, pupil constriction, blurred vision, blurred vision, muscle spasms, muscle spasms, diarrhea, slow diarrhea, slow heartbeat, and a heartbeat, and a drop in blood drop in blood pressure pressure

PSL syndrome PSL syndrome perspiration, perspiration, salivation, salivation, lachrymation lachrymation

Ibotenic acid – muscimolIbotenic acid – muscimol

Amanita Amanita muscariamuscaria

muscle muscle spasms, spasms, dizziness, dizziness, vomiting vomiting

deep deep sleep/intense sleep/intense dreamsdreams

Abbaye de Plaincourault, Mérigny, France

Psilocybin-Psilocin Psilocybin-Psilocin

hallucinogenic hallucinogenic Similar Similar

structure to structure to bufotenin and bufotenin and myristicin myristicin (nutmeg)(nutmeg)

Diverse gasterointestinal Diverse gasterointestinal toxinstoxins

AgaricusAgaricus, , AmanitaAmanita, , BoletusBoletus, , ChlorophyllumChlorophyllum, , EntolomaEntoloma, , HebelomaHebeloma, , LactariusLactarius, , MarasmiusMarasmius, , NaematolomaNaematoloma, , OmphalotusOmphalotus, , PholiotaPholiota, , RussulaRussula, , SclerodermaScleroderma, , TricholomaTricholoma,,

B. splendidusB. satanus

Tricholoma Tricholoma equestreequestre (= (= T. T. flavovirensflavovirens) )

Muscle wastingMuscle wasting fatigue, weakness fatigue, weakness

and myalgia and myalgia heart muscle heart muscle

lesions and kidney lesions and kidney lesions lesions