Elena Piecková

Adverse Health Effects of Indoor Moulds

Abstract Building associated illnesses - sick building syndrome (SBS) as a common example - are associated with staying in buildings with poor indoor air quality. The importance of indoor fungal growth in this phenomenon continues to be evident, even though no causative relation has been established so far. Indoor humidity is strongly associated with the symptoms of SBS. Fungal metabolites that may induce ill health in susceptible occupants comprise beta-D-glucan, mycotoxins, and volatile organic compounds as known irritants and/or immunomodulators. Indoor toxic fungal metabolites might be located in micromycetal propagules (endometabolites), in (bio-)aerosol, detritus, and house dust (exometabolites) as their particular carriers. It is highly probable that hyphal fragments, dust, and particles able to reach the alveoli have the strongest depository and toxic potential. Most fungal spores are entrapped by the upper respiratory tract and do not reach further than the bronchi because of their size, morphology, and the mode of propagation (such as slime heads and aggreggation). This is why studies of the toxic effects of fungal spores prefer directly applying metabolite mixtures over mimicking real exposure. Chronic low-level exposure to a mixture of fungal toxicants and other indoor stressors may have synergistic effects and lead to severe neuroendocrineimmune changes. Bolesti povezane sa stanovanjem te sindrom bolesne zgrade (engl. sick building syndrome, SBS), kao jedan od njihovih tipičnih primjera, povezani su s boravkom u zgradama s lošom kvalitetom zraka. Premda još nije uspostavljena jasna uzročno-posljedična povezanost, važnu ulogu u spomenutom fenomenu ima rast plijesni u unutarnjim prostorima. Pojavnost simptoma sindroma bolesne zgrade snažno ovisi o vlažnosti prostora. U metabolite plijesni koji mogu izazvati poremećaje zdravlja u osjetljivijih ispitanika ubrajamo beta-D-glukan, mikotoksine te hlapljive organske spojeve koji su poznati kao iritansi i/ili imunomodulatori. Toksični metaboliti gljiva unutarnjih prostora mogu biti sadržani u propagulama mikromiceta (endometaboliti), (bio)aerosolu, detritusu te kućnoj prašini (egzometaboliti). Najjači potencijal za depoziciju te toksični potencijal imaju fragmenti hifa, prašina i čestice koje mogu doprijeti do alveola. Većinu gljivičnih spora zaustave gornji dišni putovi te one zbog svoje veličine, morfologije i načina razmnožavanja (npr. ljepljive glavice i agregacija) ne dopiru dalje od bronha. To je razlog zašto se u istraživanjima toksičnih učinaka gljivičnih spora češće primjenjuju smjese metabolita negoli oponaša stvarna izloženost. Kronična izloženost niskim razinama smjesa gljivičnih toksikanata i drugih stresora unutarnjih prostora može izazvati sinergijske učinke i dovesti do teških neuroendokrino-imunosnih promjena.

Detection of ciliostatic activity in fungal growth on building materials

Varying ciliostatic activity was found in extracts of biomass from building materials inoculated with pure isolates of some molds previously isolated from buildings. Extracts from growth on materials composed of finely divided cellulose (plaster board and construction cardboard) were more active than those from growth on Rockwool. Extracts of biomass from all three materials demonstrated an activity greater than control samples. Based on the activity observed here, it is conceivable that the measurement of ciliostatic activity of biomass scraped from contaminated building materials could be useful for planning removal strategies.

Inflammatory and Haematotoxic Potential of Indoor Stachybotrys chartarum (Ehrenb.) Hughes Metabolites

Inflammatory and Haematotoxic Potential of Indoor Stachybotrys chartarum (Ehrenb.) Hughes Metabolites Mould Stachybotrys chartarum (Ehrenb.) Hughes is known to pose a health risk in indoor environments. Most of its strains can produce several intra- and extracellular trichothecene mycotoxins. Complex secondary metabolites of stachybotrys isolates from mouldy dwellings/public buildings in Slovakia were intratracheally instilled in Wistar male rats (4 μg in 0.2 mL of 0.2 % dimethylsulphoxide; diacetoxyscirpenol as the positive control). After three days, haematological parameters were measured in peripheral blood and inflammatory response biomarkers in bronchoalveolar lavage fluid (BALF), and the results were statistically analysed. Exometabolites proved to suppress red blood cell (RBC), decreasing the total RBC count, haemoglobin, and haematocrit. The exposed rats showed significantly higher total BALF cell count, indicating inflammation, lower alveolar macrophage counts, and increased granulocyte count related to the BALF cells. Due to haematotoxic and inflammation-inducing properties, metabolites of S. chartarum can cause damage to the airways and haematological disorders in occupants of mouldy buildings. Upalni i hematotoksični potencijal metabolita plijesni Stachybotrys chartarum (Ehrenb.) Hughes u zatvorenim prostorijama Plijesan Stachybotrys chartarum (Ehrenb.) Hughes poznata je kao rizični mikroorganizam u zatvorenim prostorijama. Većina njezinih sojeva može proizvesti nekoliko unutarstaničnih i izvanstaničnih trikotecenskih mikotoksina. Muškim Wistar štakorima instilirani su intratrahealno kompleksni sekundarni metaboliti stahibotrisa izolirani iz stambenih i javnih zgrada u Slovačkoj zahvaćenima plijesni (4 μg na 0,2 mL 0,2 %-tnog dimetilsulfoksida; dok se diacetoksiscirpenol rabio kao pozitivna kontrola). Tri dana kasnije izmjereni su hematološki parametri u perifernoj krvi te biopokazatelji upalnoga odgovora u bronhoalveolarnome ispirku te su rezultati obrađeni statistički. Pokazalo se da egzometaboliti suprimiraju eritrocite, smanjujući njihov ukupni broj, hemoglobin i hematokrit. Izloženi štakori imali su značajno veći broj stanica u bronhoalveolarnome ispirku, što upućuje na upalu, dok im je broj alveolarnih makrofaga bio manji, a broj granulocita povezanih sa stanicama u ispirku veći. Zbog svojih hematotoksičnih i upalnih svojstava S. chartarum može dovesti do oštećenja dišnih putova i poremećaja u krvotvornome sustavu osoba koje žive i/ili rade u zgradama zahvaćenima tom plijesni.

Respiratory toxicity of Aspergillus versicolor: the most common indoor mould in Slovakia.

A mould Aspergillus versicolor clearly dominates in damp and mouldy indoor environments under Slovak dwelling/public building conditions (up to 1/3 of all isolates during the last decade’s surveys). Nearly all of its isolates are able to synthesize a mycotoxin sterigmatocystin (detected by LC/MS-MS), that showed severe in vitro as well as in vivo toxic potential in animal experiments (after intratracheal instillation to rats). In vitro toxicity of complex chloroformextractable endoand exometabolites of 10 indoor, and related outdoor, A. versicolor isolates from a heavily mouldy kids’ fashion store in Slovakia with complaints from the occupants of irritation of their airways has been evaluated by a bioassay with tracheal organ cultures of one-day old chicks (20 microg of toxicants per mL of cultivation medium). In the in vivo experiments, respiratory toxicity of the same metabolite mixtures was tested in Wistar rats during three days. The inflammatory and cytotoxic biomarkers were then analyzed in bronchoalveolar lavage fluid. Searching for the fungus possible source, molecular epidemiological study of the isolates was performed using RAMP PCR. Strains colonizing the indoor walls of the shop were the highest correlated to the outdoor airborne ones (Pearson correlation 97%). While indoor airborne isolates correlated to the strains growing on retailed clothes at the levels of 90 or 86% according to Pearson. All micromycetes produced secondary metabolites that ceased ciliary beating in tracheal epithelium in the organ cultures already in 24 hrs of the activity, i.e. in the sense of the method used, they belong to strong toxicants. Two of the isolates tested also produced extrolites without toxic effects detectable by the method. The metabolites also showed certain cytotoxic Environmental Health and Biomedicine 135 www.witpress.com, ISSN 1743-3525 (on-line) WIT Transactions on Biomedicine and Health, Vol 15,

In Vitro Toxicity Of Indoor Fungi From DwellingsIn Slovakia: Testing On The Isolated Lung Cells

The lung is the target organ of multiple aggressions due to environmental noxious substances, indoor pollution, occupational hazards and personal risk such as cigarette smoke. Metabolites produced by different fungus species can also be present the inhaled air. Their effects are not frequently studied in the lung or in the lung cells. Our study focused on the effects of metabolites (both exoand endometabolites) produced by Aspergillus ustus, Aspergillus versicolor, Penicillium chrysogenum and Stachybotrys chartarum isolated from dwellings in Slovakia. Their effects were studied in vitro on alveolar macrophages and epithelial type II cells isolated from Wistar rats and Clara cells isolated from mice. Alveolar macrophages represent a free living population in the alveolar spaces and play an important role in maintaining clean and sterile alveoli; they contribute also to the production of proinflammatory cytokines. Epithelial type II cells play a critical role in preserving the functional integrity of the alveolar surface and they also produce cytokines. The effects of metabolites were evaluated by estimating their cytotoxicity, the activity of lysosomal enzyme acid phosphatase in alveolar macrophages. Lectins were used for studying the changes on cell surface. The production of cytokines (Monocyte Chemoatractant Protein 1MCP-1 and Tumor Necrosis Factor α – TNF-α) was also measured. The effects of metabolites were dose dependent, the highest toxicity was evoked by Stachybotrys chartarum metabolites.

Use of the in vitro model for the evaluation of toxic effects of metabolites produced by fungi

There are several reports about chronic intoxications, allergies or other pulmonary health problems of dwellers (especially babies) and workers from buildings contaminated with microfungi. Material from presented work was collected during the study of indoor fungal colonization of mouldy dwellings and public buildings in Slovakia. The isolated fungi were cultivated for 10 days in liquid media and their products - both endo- and exometabolites were isolated. The metabolites from four isolates of Stachybotrys chartarum, two of Aspergillus versicolor and one of Penicillium chrysogenum were used. Their effects were studied in vitro on rat alveolar epithelial type 2 cells, from the toxicological point of view one of the most important lung cells type. Type 2 cells were cultured for 20 hours in DMEM and then they were exposed for 24 hours to various concentrations of metabolites. The toxicity was evaluated by staining the cells for alkaline phosphatase which is a marker for this cell type. The toxic effect of metabolites from all isolates of Stachybofrys chartarum was dose dependent, there were differencies in the extent of cytotoxicity. From the tested microorganisms Stachabotrys chartarum and Aspergillus versicolor showed high toxic effects (Stachybotrys higher than Aspergillus), • while Penicillium chrysogenum only minor toxic effects in this test.

Sterigmatocystin in foodstuffs and feed: aspects to consider

ABSTRACT Sterigmatocystin (STC) is a possible human carcinogen (2B) according to International Agency for Research on Cancer classification and has been associated with immunotoxic and immunomodulatory activity, together with mutagenic effects. It might be found in numerous substrates, from foods and feeds to chronically damp building materials and indoor dust. Although European Food Safety Authority concluded that the exposure to STC to be of low concern for public health, reinforces the need of data concerning exposure of European citizens. Climate change can represent an increased risk of exposure to STC since it is a crucial factor for agro-ecosystem powering fungal colonisation and mycotoxin production This aspect can represent an increased risk for European countries with temperate climates and it was already reported by the scientific community.

Indoor Microbial Aerosol and Its Health Effects: Microbial Exposure in Public Buildings – Viruses, Bacteria, and Fungi

Mechanisms of aerosolization of microorganisms, composition and dynamics of microbioaerosol are characterized. As well as methods of its detection, incl. modern equipment set-ups and sampling procedures recommended are outlined. Medical impact of (indoor) air disperged viral, bacterial and fungal propagules (allergies, intoxications, infections), together with the related European legislation is summarized. An overview of real mycoaerosol conditions in our dwellings and their outdoors with different microclimate, settlement and building types, household characteristics and health state of occupants is given, too. Finally, examples of several possible health damages due to exposition to (aerosolized) fungal toxicants in vitro and in vivo are demonstrated.

Adverse Health Effects Of Indoor Moulds / Štetni Zdravstveni Učinci Plijesni Unutarnjih Prostora

Abstract Building associated illnesses - sick building syndrome (SBS) as a common example - are associated with staying in buildings with poor indoor air quality. The importance of indoor fungal growth in this phenomenon continues to be evident, even though no causative relation has been established so far. Indoor humidity is strongly associated with the symptoms of SBS. Fungal metabolites that may induce ill health in susceptible occupants comprise beta-D-glucan, mycotoxins, and volatile organic compounds as known irritants and/or immunomodulators. Indoor toxic fungal metabolites might be located in micromycetal propagules (endometabolites), in (bio-)aerosol, detritus, and house dust (exometabolites) as their particular carriers. It is highly probable that hyphal fragments, dust, and particles able to reach the alveoli have the strongest depository and toxic potential. Most fungal spores are entrapped by the upper respiratory tract and do not reach further than the bronchi because of their size, morphology, and the mode of propagation (such as slime heads and aggreggation). This is why studies of the toxic effects of fungal spores prefer directly applying metabolite mixtures over mimicking real exposure. Chronic low-level exposure to a mixture of fungal toxicants and other indoor stressors may have synergistic effects and lead to severe neuroendocrineimmune changes. Bolesti povezane sa stanovanjem te sindrom bolesne zgrade (engl. sick building syndrome, SBS), kao jedan od njihovih tipičnih primjera, povezani su s boravkom u zgradama s lošom kvalitetom zraka. Premda još nije uspostavljena jasna uzročno-posljedična povezanost, važnu ulogu u spomenutom fenomenu ima rast plijesni u unutarnjim prostorima. Pojavnost simptoma sindroma bolesne zgrade snažno ovisi o vlažnosti prostora. U metabolite plijesni koji mogu izazvati poremećaje zdravlja u osjetljivijih ispitanika ubrajamo beta-D-glukan, mikotoksine te hlapljive organske spojeve koji su poznati kao iritansi i/ili imunomodulatori. Toksični metaboliti gljiva unutarnjih prostora mogu biti sadržani u propagulama mikromiceta (endometaboliti), (bio)aerosolu, detritusu te kućnoj prašini (egzometaboliti). Najjači potencijal za depoziciju te toksični potencijal imaju fragmenti hifa, prašina i čestice koje mogu doprijeti do alveola. Većinu gljivičnih spora zaustave gornji dišni putovi te one zbog svoje veličine, morfologije i načina razmnožavanja (npr. ljepljive glavice i agregacija) ne dopiru dalje od bronha. To je razlog zašto se u istraživanjima toksičnih učinaka gljivičnih spora češće primjenjuju smjese metabolita negoli oponaša stvarna izloženost. Kronična izloženost niskim razinama smjesa gljivičnih toksikanata i drugih stresora unutarnjih prostora može izazvati sinergijske učinke i dovesti do teških neuroendokrino-imunosnih promjena.

Štetni zdravstveni učinci plijesni unutarnjih prostora

Abstract Building associated illnesses - sick building syndrome (SBS) as a common example - are associated with staying in buildings with poor indoor air quality. The importance of indoor fungal growth in this phenomenon continues to be evident, even though no causative relation has been established so far. Indoor humidity is strongly associated with the symptoms of SBS. Fungal metabolites that may induce ill health in susceptible occupants comprise beta-D-glucan, mycotoxins, and volatile organic compounds as known irritants and/or immunomodulators. Indoor toxic fungal metabolites might be located in micromycetal propagules (endometabolites), in (bio-)aerosol, detritus, and house dust (exometabolites) as their particular carriers. It is highly probable that hyphal fragments, dust, and particles able to reach the alveoli have the strongest depository and toxic potential. Most fungal spores are entrapped by the upper respiratory tract and do not reach further than the bronchi because of their size, morphology, and the mode of propagation (such as slime heads and aggreggation). This is why studies of the toxic effects of fungal spores prefer directly applying metabolite mixtures over mimicking real exposure. Chronic low-level exposure to a mixture of fungal toxicants and other indoor stressors may have synergistic effects and lead to severe neuroendocrineimmune changes. Bolesti povezane sa stanovanjem te sindrom bolesne zgrade (engl. sick building syndrome, SBS), kao jedan od njihovih tipičnih primjera, povezani su s boravkom u zgradama s lošom kvalitetom zraka. Premda još nije uspostavljena jasna uzročno-posljedična povezanost, važnu ulogu u spomenutom fenomenu ima rast plijesni u unutarnjim prostorima. Pojavnost simptoma sindroma bolesne zgrade snažno ovisi o vlažnosti prostora. U metabolite plijesni koji mogu izazvati poremećaje zdravlja u osjetljivijih ispitanika ubrajamo beta-D-glukan, mikotoksine te hlapljive organske spojeve koji su poznati kao iritansi i/ili imunomodulatori. Toksični metaboliti gljiva unutarnjih prostora mogu biti sadržani u propagulama mikromiceta (endometaboliti), (bio)aerosolu, detritusu te kućnoj prašini (egzometaboliti). Najjači potencijal za depoziciju te toksični potencijal imaju fragmenti hifa, prašina i čestice koje mogu doprijeti do alveola. Većinu gljivičnih spora zaustave gornji dišni putovi te one zbog svoje veličine, morfologije i načina razmnožavanja (npr. ljepljive glavice i agregacija) ne dopiru dalje od bronha. To je razlog zašto se u istraživanjima toksičnih učinaka gljivičnih spora češće primjenjuju smjese metabolita negoli oponaša stvarna izloženost. Kronična izloženost niskim razinama smjesa gljivičnih toksikanata i drugih stresora unutarnjih prostora može izazvati sinergijske učinke i dovesti do teških neuroendokrino-imunosnih promjena.