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Detailed Description of ERMI Species

An Assured Bio Labs White Paper Contributions were made by all of the following: James Campbell, Ph.D., Senior Scientist Alisha Coffey, B.S., Molecular Analyst

Project Director Edward A. Sobek, Ph.D., Chief Science Officer

March 3, 2008

Table of Contents Introduction 3 Scopulariopsis brevicaulis 11 Problem Statement 3 Scopulariopsis chartarum 11 Previous Options 3 Stachybotrys chartarum 11 The Assured Bio Solution 3 Trichoderma viride 11 Implementation 3 11 Summary 3 Alternaria alternata 4 flavus 4 Aspergillus fumigatus 4 Aspergillus niger 4 Aspergillus ochraceus 5 Aspergillus penicilloides 5 Aureobasidium pullulans 5 Aspergillus restrictus 5 Aspergillus sclerotiorum 5 Acremonium strictum 5 Aspergillus sydowii 5 Aspergillus unguis 6 Aspergillus ustus 6 Aspergillus versicolor 6 cladosporioides 6 Chaetomium globosum 6 7 Cladosporium sphaerospermum 7 Eurotium amstelodami 7 Epicoccum nigrum 8 amphibiorum 8 Penicillium brevicompactum 9 9 9 9 10 10 10 Paecilomyces variotii 10 Rhizopus stolonifer 11

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008

Benefit 2 Introduction Mycological lineages were analyzed to The Environmental Relative ensure that ecological descriptions are Moldiness Index (ERMI) contains 36 traceable to each ERMI species or species species specific DNA probes. Mold group. Specific Quantitative PCR (MSQPCR) is the method used to detect and quantify the ERMI mold Benefit 3 species. MSQPCR and ERMI were All descriptions will be updated quarterly. developed via an intense research Assured Bio’s current ERMI/MSQPCR will project conducted over a period of 10 be granted access to each updated version years at the United States EPA. electronically.

Implementation Problem Statement ERMI/MSQPCR clients may freely use the There is no definitive source that species descriptions to enhance their own describes the ecology and/or medical reports. The descriptions can be pasted significance of the mold (fungal) directly into their own inspection reports. species contained in the ERMI panel. Inspectors can also attach the verified electronic version as an appendix to their reports. Previous Options Indoor air quality investigators had Current users of Assured Bio’s ERMI and to comb through old journal articles, other MSQPCR panels will automatically out-of-print mycology keys and receive a high-quality glossy hard copy of texts, plus spend vast hours on the the species description. New clients or internet in order to find information clients ordering their first ERMI test will on the species of that were automatically receive one high quality print present in the ERMI report. copy via mail. A verified electronic version Unfortunately, much of the internet will always accompany ERMI/MSQPCR data cannot be verified and many panel reports. In addition, current species names have changed over the ERMI/MSQPCR clients will receive a high last decade. quality print with each quarterly update.

The Assured Bio Solution Summary Provide an up to date description of The vast wealth of data in the ERMI and all species and species groups other MSQPCR panels, coupled with present in the ERMI test panel definitive and detailed species descriptions, will considerably enhance the interpretation of ERMI/MSQPCR panels for the inspector Benefit 1 and significantly improve discussion of the All species descriptions are from results among clients and remediation peer-reviewed journals or up-to-date specialists, medical professionals, and medical mycology texts. litigation attorneys.

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008

Aaltr Afumi Alternaria alternata Aspergillus fumigatus / Neosartorya fischeri This can be found throughout Neosartorya fischeri is a heat-tolerant the world on and in plants, soils, fungus that is common in soil and fruits and textiles and foods. A. alternata is occasionally causes human infections. among the most commonly observed Aspergillus fumigatus is also heat-tolerant molds in indoor environments, and and found worldwide. A. fumigatus it is its are released diurnally. It is particularly dense in agricultural soils but is found frequently in moist/humid commonly isolated from house dust, areas such as watertanks and garbage, compost, potted plants, humidifiers humidifiers. However, this species and HVAC systems, as well. More is also quite common in dry areas importantly, A. fumigatus is isolated including dust from floors and commonly from human patients. In healthy mattresses. This species produces humans, A. fumigatus is not a . several and However, this species can cause severe (tenuazonic acid and altertoxins). infections in humans with suppressed Members of the Alternaria are immune systems (e.g. those with pre- known to cause asthma, sinusitis and existing illnesses or taking infections of the eyes, ears and skin. immunosuppressants). In such individuals, spores that are inhaled are not attacked Aflav efficiently by the host’s immune system, and / oryzae the spores could germinate and begin to Isolates of Aspergillus flavus and A. invade host tissues. oryzae are morphologically indistinguishable. A. flavus can be Anigr found virtually anywhere on Earth Aspergillus niger / awamori / foetidus / and has been isolated from dry areas phoenicis in Chile, antarctic lakes, humidifiers, Species detected by this assay are plants, insects, animals, leather, morphologically similar and difficult to feathers, cotton fabrics, paintings, distinguish without molecular techniques, etc. Distributions of A. flavus in such as PCR. Aspergillus awamori is American soils are more dense in the widespread in soils and on plants, and it has southern U.S. A. flavus can produce been used extensively for industrial aflatoxins under some circumstances. applications and for food preparation. Some Aflatoxin B1 is the most potent isolates of A. awamori have been found to carcinogen (cancer-causing agent) produce the known as ochratoxin known, and lethal doses of this A, and it is possible that this fungus can compound are known to be cause subcutaneous infections. A. foetidus extremely low in mice. In man, A. and A. phoenicis are soil fungi that are likely flavus can cause pulmonary involved in natural decomposition. A. niger and weakened patients is a fungus that can be found in house dust, can suffer from cutaneous, nasal and mattress dust. A. niger can also contaminate cerebral infections. foods such as spices and onions. Importantly, A. niger is allergenic and can

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 cause inner/outer ear infections and sinus infections. Aochr1 Arest Aspergillus ochraceus / ostianus Aspergillus restrictus / caesillus / conicus These species of Aspergillus can be Aspergillus restrictus is a fungus that is found indoors and on foodstuffs (e.g. more likely to be isolated in cool and dry coffee and paprika). Both species climates, which could explain its frequent produce ochratoxin A, but A. occurrence in house dust. All three of these ostianus can also produce aflatoxin. species are considered medically important, although infections are not widely Apeni2 documented. Aspergillus penicillioides This fungal species is common in Asclr very dry conditions and can be Aspergillus sclerotiorum isolated from dried fruits, spices, This species is found in tropical and archives, furniture, carpets, house subtropical soils across the world. A. dust and clothing. It is also sclerotiorum can produce ochratoxins and is associated with dust mites and is known to cause infections of the ear, known to be allergenic. toenails and fingernails.

Apull Astrc Aureobasidium pullulans Acremonium strictum This fungal species is ubiquitous. This species is a common inhabitant of soils Isolations are most common from worldwide and can be isolated from plant plant leaves but have been successful surfaces, fuel and fuel filters. Import in an from such diverse environments as indoor context, A. strictum is found widely humidifiers, house dust, mattress in the atmosphere and is common observed dust, forest soils, sand dunes, peat on food and moist indoor surfaces (e.g. bogs, estuarine sediments, marine humidifiers). It is possible that moldy sediments and seawater. In British homes can show greater numbers of this homes, airborne spores of this species in winter months. A. strictum has species increase sharply in winter caused infections in chemotherapy and months. Interestingly, A. pullulans transplant patients. Infections of blood, does not appear to require high levels cerebrospinal fluid, eye, pulmonary, of nutrients commonly needed by peritoneal, toenail and fingernail have been other environmental microbes. This reported for this species, although they species is also extremely sensitive to appear to be relatively rare. However, heat, and can be found in high- reports of infections by species of the genus humidity areas (e.g. window frames Acremonium appear to be on the rise. and bathrooms). This fungus is implicated rarely in human infections Asydo3 of the eyes and skin, and A. pullulans Aspergillus sydowii infections can be found in blood. This species is found in soils worldwide and has been isolated from plants, seeds, foods, leather, textiles and uranium mines. A. sydowii can produce mycotoxins know as

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 sydowic acids and can cause allergenic, and mycoses of this species fingernail and toenail infections and include osteomyelitis and infections of the invasive aspergillosis. auditory canal, fingernails and toenails. Aungu Aspergillus unguis Cclad1 and Cclad2 Very little is known about this fungal Cladosporium cladosporioides svar. 1 and species. However, it has been found Cladosporium cladosporioides svar. 2 to cause fingernail and toenail These two organisms are not currently infections. recognized as individual species, and they cannot be differentiated using standard Austs2 microscopic techniques. DNA sequencing Aspergillus ustus projects seeking to devise rapid It is likely that A. ustus is one of the identification methods for fungal most widely spread species of detected distinct DNA sequences in this Aspergillus. It has been isolated species, and each is now recognized as a from diverse soils from around the “sequevar.” In essence, they cannot be world, salt marshes, estuaries, foods, identified correctly without the use of DNA- bat caves and uranium mines. based technology, such as this quantitative Sporulation of A. ustus is stimulated PCR technique. Both sequevars represent by light. This species produces the most common saprobe in the several mycotoxins and has been environment. This species generates many responsible for endocarditis and more spores under moist conditions than in infections of the and skin. It is dry conditions. C. cladosporioides is possible that infection by A. ustus is distributed worldwide in soils, air, house nosocomial, but diagnoses of this dust, mattress dust, on dairy products, are rare. textiles, food, plants, many aquatic environments, wood pulp and feathers. This Avers2-2 species is allergenic and can form fungal Aspergillus versicolor balls in lungs, skin infections, keratitis, As are most aspergilli, A. versicolor sinusitis, and infections of spinal fluid, is extremely widespread in nature. fingernails and toenails. However, this species tends to occupy the coldest regions of Cglob Aspergillus distributions, as well as Chaetomium globosum deserts, peat bogs, estuarine This fungus is isolated commonly from soil, sediments, compost, linoleum, decaying plants, seeds, food, estuarine chipboard, paintings, , spices, environments and marine sediments. It has stored grains, house dust, mattress particular notoriety as a soft rot fungus and dust and rotting military equipment can be found on decaying wood, explaining in the tropics. This species is its occurrence in indoor environments extremely xerophilic and common in following water damage. In fact, C. indoor environments, where its globosum can be found growing on growth can cause moldy odors. A. wallpaper in homes with extensive water versicolor is known to produce a damage. Sporulation of this fungus tends to carcinogenic compound known as occur more readily under dark conditions, sterigmatocystin. A. versicolor is and the spores produced are very resistant to

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 desiccation. While not particularly them. C. sphaerospermum is one of the allergenic itself, its presence appears most commonly isolated indoor air fungi. to enhance the allergic response of This species is allergenic and has caused individuals to other allergens (e.g. documented bronchial lesions and pollen). This species has caused subcutaneous skin infections. invasive infections, subcutaneous infections and Eamst fingernail and toenail infections. Eurotium (Aspergillus) amstelodami / The genus Chaetomium appears to chevalieri / herbariorum / rubrum / repens be emerging as important fungal This assay identifies a group of closely pathogens. related Eurotium species. Most molds isolated from indoor environments are Cherb asexual species, however some also Cladosporium herbarum reproduce sexually. To discern these two This is a very common fungus in modes or reproductive states, mycologists nature, and it can be isolated from have devised a unique terminology. The dead/dying plants, soil, food, wheat, term “anamorph” describes those molds that textiles, floor dust, mattress dust, reproduce asexually; whereas, the term seawater, uranium mines and paint. “teleomorph” describes molds that In fact, it is possible that C. reproduce sexually. Anamorphic or asexual herbarum is the most common molds do not need a partner to reproduce, Cladosporium in air samples and they produce their spores similar to budding appears to be more prevalent in cells and do so on a grand scale; summer months in British homes. millions if not billions of spores are However, this species was found to produced in a short period of time (24-48 cause food spoilage at refrigeration hrs). Anamorphic reproduction is an temperatures. C. herbarum was evolutionary strategy that fires the conflict found to be strongly allergenic and between humans and molds in homes and produces an endotoxin that has buildings; just add water to building similar health effects to that materials, and mold will seem to appear out produced by Stachybotrys of nowhere and rapidly colonize the damp chartarum. substrates. Teleomorphic molds, however, must find and fuse with a compatible partner Cspha or strain in order to produce spores sexually. Cladosporium sphaerospermum Hence, teleomorphic molds are rare relative As with most species of to anamorphic molds because the paring of Cladosporium, C. sphaerospermum compatible strains in the environment is is common worldwide. This species governed by the laws of probability, and the can be isolated from plants, soil, probability of two microscopic strains food, paint, textiles, insulation, floor meeting at any given location is remote. dust, mattress dust, humidifiers and However, some teleomorphic species tend to from humans and other animals. commonly occur indoors. Spores of this species are difficult to distinguish from those of C. The most common teleomorphic genus is cladosporioides microscopically, but Eurotium. Eurotium species are perhaps the DNA analyses easily distinguish most abundant sexually reproducing molds

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 found indoors. The key to is not known as a pathogen, but this species Eurotium's success lies in genetics, can cause skin allergies. for Eurotium's asexual counterpart is Aspergillus. Aspergillus species produce enormous flushes of spores. Muc1 Hence, Aspergillus spores are Mucor amphibiorum / circinelloides / extremely common, especially in a hiemalis / indicus / mucedo / racemosus / water compromised building. The ramosissimus and Rhizopus azygosporus / relative abundance of Aspergillus homothalicus / microsporus / oligosporus / spores dramatically increases the oryzae probability that two compatible The species of mold represented in this aspergilli strains will meet and fuse assay are all members of a broad class of to form a teleomorphic Eurotium fungi known as zygomycetes. Zygomycetes species. Thus, Eurotium has become are primitive but fast growing fungi. They an important mold genus, one that are widely distributed in terrestrial should not be ignored during indoor environments, where they break down plant air quality assessments. This genus debris in soil. However, many species are is xerophilic and has the ability to common environmental contaminants that germinate and colonize substrates can cause food spoilage, and a few are having minimal water activity. pathogens of plants, insects and humans. By Eurotium is also a common food definition, all pathogenic zygomycotic spoilage organism. species will grow at 37°C, with the possible exception of the M. circinelloides. Eurotium has been implicated in several health maladies. The common genera that infect humans Anamorphic forms of Eurotium include Rhizopus, followed by Mucor, produce various mycotoxins. Rhizomucor, Absidia, Cunninghamella and Farmer's lung disease (FLD) is Syncephalastrum. Underlying diseases in caused mainly by repeated exposure humans include cancer and leukemia, to moldy hay colonized by Eurotium or prednisone use, diabetes, species. Eurotium may be a deferoxamine and desferrioxamine therapy, respiratory allergic in susceptible transplantation, burn wounds and the individuals and can cause adverse associated forms of immunosuppressive health effects in children who attend therapies. The most common clinical form school in buildings damaged by of is rhinocerebral disease moisture. followed by pulmonary, cutaneous/subcutaneous, gastrointestinal and Enigr disseminated disease. Mucor amphibiorum Epicoccum nigrum has not been reported in human infections. One of the most commonly isolated Mucor circinelloides has been reported as a indoor fungi, E. nigrum is also rare cause of cutaneous infections in widely distributed in nature. It can humans. Mucor hiemalis has been reported be found growing in and on soils, from a few cases of human cutaneous sand, dead/decaying plant tissue, infection. (synonym: M. saline environments, textiles and rouxii) has been reported from human moldy paper. At this time, E. nigrum gastric and pulmonary infections, a case of

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 necrotizing fasciitis and reports of implicated widely in human disease. hepatic infection in a bone marrow However, P. brevicompactum has been transplant recipient who had ingested isolated from a dog with fungal pneumonia contaminated medicine. Mucor and a deep organ infection in a human. racemosus has been infrequently reported as a causative agent of Pchry animal and human zygomycosis. Penicillium chrysogenum Rhizopus microsporus accounts for This species is found worldwide but has 10-15% of reported human cases and earned most notoriety from its production of has been implicated in cellulitis, . In addition to soil distributions, it cutaneous infection, zygomycosis, can be isolated from foods, plants, floor and gastrointestinal infections. dust, mattress dust, wood, wall paper, paint, However, rhinocerebral forms of R. gypsum (as in wall board) artwork and microsporus are rare. Rhizopus occasionally optical lenses. It is considered oryzae (synonym: R. arrhizus) is the a good indicator of water intrusion. most common causative agent of Although this species is highly allergenic zygomycosis, accounting for 60% of and can produce mycotoxins, P. the reported culture positive cases chrysogenum is not considered a common and nearly 90 percent of the health risk. Nonetheless, infections of the rhinocerebral form of infection. ears, eyes, heart tissue, skin and cerebrospinal fluid have been documented. Pbrev Penicillium brevicompactum / Pcory stoloniferum Penicillium corylophilum P. stoloniferum is a relatively rarely This species is widely distributed, but it is occurring fungus found in soils and found more frequently in warm climates. foods. P. stoloniferum commonly Isolations have been successful from soil, attacks poinsettias in Switzerland textiles and various foods. This species is greenhouses but is not currently thought to be relatively xerophilic and is recognized as a health threat. P. likely more common in low-humidity brevicompactum is a common conditions, probably explaining their species worldwide and indoors, isolation from wood and paint. At this time, occurring in fruit juices, fresh herbs, P. corylophilum does not appear to be a wall paper, wood, paint, potted human pathogen. plants (particularly strong association), soils, floor dust, PenGrp2 mattress dust, caves, freshwater and Penicillium crustosum / camembertii / uranium mines. P. brevicompactum commune / echinulatum / solitum can be xerophilic but sensitive to P. crustosum is a common food high-salt conditions. This species contaminant, particularly common in seeds, also inhibits the growth of several nuts and apples. P. crustosum produces species of soil bacteria, possibly potent neurotoxins (penitrems and through production of its several roquefortine) that can cause muscular mycotoxins (e.g. ochratoxin). P. tremors in individuals eating contaminated brevicompactum can be strongly foods. P. camemberti is a mold commonly allergenic, but it has not been found in cheeses ( ) and

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 occasionally meats, where it can aggressively on computer diskettes in high produce low levels of the mycotoxin humidity. P. glabrum also grows well on cyclopiazonic acid. P. commune is the corks of wine bottles and elicits allergic commonly found indoors and on responses in individuals that work with wine cheeses and meats. P. commune has corks. P. lividum is a relatively rare and been documented in pulmonary non-pathogenic species of Penicillium and infections in dogs and can produce occurs mostly in northern latitudes. P. cyclopiazonic acid and possibly purpurescens is a common inhabitant of nephrotoxins. P. echinulatum is soils and indoor environments (particularly found most frequently on foods greenhouses). P. purpurescens does not containing oils (e.g. margarine and appear to be an overt pathogen, but it can be cheese) but is also found indoors. P. found in feed potentially toxic to poultry. P. echinulatum is capable of producing spinulosum is distributed worldwide and is tremorgenic mycotoxins (territrems). usually found associated with forest soils, P. solitum is commonly isolated flour-based foods and fruit products. P. from foods such as hard cheeses and spinulosum can grow on wet plasterboard, some meats. P. solitum can produce and such growth can yields mycotoxin mycotoxins (viridicatins) on such production, the health effects of which are foods but does not appear to cause under debate. P. thomii is widely distributed diseases in humans. in soils of temperate environments. P. thomii does not appear to be pathogenic, Ppurp given current data. Penicillium purpurogenum This is another example of a Pvarb2 Penicillium with a worldwide Penicillium variabile distribution in soils. This species This species is widely distributed in soils also occurs on foods, plants and and can also be found in seawater, fruit occasionally on optical lenses. P. juices, paper and optical lenses. P. variabile purpurogenum tends to grow in appears to grow best at slightly acidic pH environments with low pH (acidic). and does not tolerate high heat for long A mycotoxin, known as rubratoxin, periods of time. This species produces can be produced when growth occurs ochratoxin A (among others) but is not on foods. P. purpurogenum is not currently known as a pathogen. currently recognized as a pathogen, but it has caused a few pulmonary Pvari2 infections in humans and a systemic Paecilomyces variotii infection in a dog. This fungus is known to be heat resistant and can, therefore, be found most commonly Pspin2 in warm and arid environments. It is also Penicillium glabrum / lividum / very common in air, animal feed, seawater, purpurescens / spinulosum / thomii wood pulp in paper mills, creosote-treated P. glabrum is a commonly occurring wood, walls, wallpaper, house dust, indoor fungus, but it can also be compost, leather, optical lenses, synthetic found contaminating foods rubber, photographic paper, moldy cigars, (particularly fruit and fruit products) ink, optical lenses, PVC and kerosene. P. and growing in compost and variotii has been known as a pathogen in

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 birds and mammals but also appears brevicaulis, this species produces infections to be an important human pathogen of the skin and fingernails and toenails. and infects the heart, lungs, bones, spleen and soft tissue. SCchr Scopulariopsis chartarum Rstol Relatively little is known about Rhizopus stolonifer Scopulariopsis chartarum, not to be This fungus has a worldwide confused with Stachybotrys chartarum. distribution, occurring most densely Scopulariopsis chartarum was first observed in soils of warm climates. R. on wallpaper, but has also been found in stolonifer is one of the most soils. Growth on maple by this species frequently observed indoor air fungi results in a weakening of the wood. This and commonly grows on foods (e.g. species does not appear to be a human bread) and its spores can germinate pathogen, but it has caused a systemic on moist paper. It appears that mycosis in a dog. growth is enhanced by slightly alkaline conditions. This species has Stac caused occasional infections, but it is Stachybotrys chartarum not generally regarded as an Stachybotrys chartarum is the quintessential important pathogen. black mold found in indoor environments. It is distributed worldwide, primarily found SCbrv associated on decaying plant material. S. Scopulariopsis brevicaulis / fusca chartarum possesses a battery of S. brevicaulis is the most common linked to plant decomposition, making it a species of its genus and occurs potent attacker of all forms of wood, paper worldwide and occurs in soils, floor and natural fibers (e.g. wool). Hence, it is dust, mattress dust, aquatic commonly an indicator of moisture environments, compost, seawater, problems in homes and can be found paper mill waste, wood pulp, textiles, growing on paper, wallpaper, wall board, paintings and uranium mines. S. wood and textiles. S. chartarum is not a fusca is also commonly isolated from common pathogen, in and of itself, but has soil, straw, paper and food. S. garnered particular attention for its role in brevicaulis is regarded as moderately Sick Building Syndrome, due to its high xerophilic, and it can produce toxic production of mycotoxins (satratoxin G and by-products of arsenic and mercury, H). Long-term exposure to such toxins can becoming exceptionally dangerous induce a myriad of health maladies, when growing indoors on paints including nausea, dermatitis, rhinitis, containing arsenic. S. brevicaulis is depression, general malaise, headaches, sore said to produce - or ammonia- throats, etc. S. chartarum has also been like odors when growing indoors. S. known to invade lung tissue. brevicaulis attacks and keratin, often leading to infections of the Tviri toenails and fingernails. However, it Trichoderma viride / atroviride / koningii can also cause skin, lung and soft T. viride and T. koningii are cosmopolitan tissue infections. S. fusca is less species and have been isolated from almost frequently pathogenic than S. every environment. Soils, composts and

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[ERMI/MSQPCR PANEL SPECIES DESCRIPTION] March 12, 2008 vegetables are common sources of these fungi, and cool and moist environments are preferred. Very little is known about T. atroviride. T. viride can grow on linoleum and wallpaper, and is probably more commonly isolated from indoor environments in winter months. As a genus, Trichoderma can cause nosocomial (hospital acquired) mycoses from contaminated solutions. T. viride is allergenic and has caused keratitis, peritonitis, pulmonary infections and hematomas.

Wsebi Wallemia sebi This fungus is a very common indoor fungus and is commonly found airborne. It is xerophilic and osmophilic and can be found growing on substrates that would desiccate many other fungi. These substrates include rock salt, bacon, salted foods, jam, jellies, fruits, textiles, rotting paper, and mammals. W. sebi can also be found in floor dust, mattress dust, soil and hay. This species is allergenic and is known to colonize human lungs, bones and skin. However, W. sebi is not considered a serious pathogen.

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