Z problemem grzybów degradujących materiały budowlane człowiek styka się od zarania dziejów. Bardzo wcześnie zdano sobie sprawę, iż obecność tych organizmów w pomieszczeniach wpływa szkodliwie na zdrowie przebywających w nich ludzi. Jednym z pierwszych objawów świadczących o obecności mikroorganizmu jest zapach unoszący się we wnętrzu. Wskaźnik ten wydaje się o tyle istotny, że wyczuć go można często, zanim uda się makroskopowo dokładnie określić miejsce infekcji. Obecnie przeważa opinia, że zawarte w powietrzu związki zapachowe, zwane lotnymi związkami organicznymi VOC (ang. – volatile organic compounds), wydzielane przez różne mikroorganizmy przyczyniają się do tak zwanego syndromu chorego budynku – SBS (ang. – sick building syndrome). Grzyby wytwarzają szerokie spectrum związków organicznych, wśród których najważniejsze są: alkohole, estry, ketony, mono- i seskwiterpeny. Stwierdzono różnice w ich wydzielaniu zarówno w przypadku poszczególnych gatunków, jak i szczepów. Okazało się, iż podłoże oraz warunki środowiska, w których wzrastają grzyby, wpływają na emisję mikrobiologicznych lotnych związków organicznych.
Przeprowadzone badania wykazały, iż istnieje zależność między rodzajem materiału budowlanego zasiedlonego przez mikroorganizm a oznaczanymi w atmosferze lotnymi związkami organicznymi. Jeden ze sposobów ograniczenia rozkładu drewna i towarzyszącemu zjawiska emisji lotnych związków organicznych polega na stosowaniu chemicznych środków ochrony drewna. Jednak czasami zdarza się, że chociaż drewno zostało nasycone fungicydami, nadal jest podatne na porastanie przez grzyby degradujące tkankę drzewną. Zjawisko to wynika z niedostatecznej retencji fungicydu w drewnie lub powstaniu szczepu mikroorganizmu odpornego na chemiczny środek ochrony drewna w zastosowanej koncentracji. Nie przeprowadzono do tej pory badań nad emisją VOC zwiazaną z rozwojem grzybów rosnących na drewnie zabezpieczonym dawką podprogową fungicydu.

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1. WSTĘP

2. GENEZA PROBLEMU

3. CEL PRACY

4. MATERIAŁ BADAWCZY I METODYKA
4.1. Grzyby testowe 
4.2. Drewno
4.3. Fungicydy 
4.4. Karbaminian-3-jodo-2-propynylbutylowy (IPBC)
4.5. Propikonazol 
4.6. Chlorek didecylodimetyloamoniowy (DDAC)
4.7. Metoda oznaczania lotnych związków organicznych 

5. WYNIKI BADAŃ I DYSKUSJA
5.1. Lotne związki organiczne wydzielane przez podłoże 
5.2 Izolacja lotnych związków organicznych w zależności od rodzaju fazy stacjonarnej adsorberów
5.3. Oznaczenie VOC wydzielanego przez grzyby porastające drewno 
5.4. Dynamika tworzenia lotnych metabolitów przez Aspergillus niger van Tieghem porastającego zaimpregnowane drewno
5.5. Oznaczenie VOC wydzielanego przez Trichoderma viride Persoon ex S.F. Gray aggr. 
5.6. Dynamika tworzenia związków lotnych przez Trichoderma viride Persoon ex S.F. Gray aggr. 
5.7. Dynamika tworzenia związków lotnych przez Trichoderma viride Persoon ex S.F. Gray aggr porastającego zaimpregnowane drewno
5.8. Oznaczenie VOC wydzielanego przez Coniophora puteana (Schumacher ex Fries) Karsten 
5.9. Dynamika tworzenia związków lotnych przez Coniophora puteana (Schumacher ex Fries) Karsten porastającego zaimpregnowane drewno
5.10. Oznaczenie VOC wydzielanego przez Gloeophyllum trabeum (Persoon ex Fries) Murrill porastającego podłoże zaimpregnowane 
5.11. Dynamika tworzenia lotnych metabolitów przez Gloeophyllum trabeum (Persoon ex Fries) Murrill porastającego zaimpregnowane drewno 
5.12. Wykrywanie ognisk korozji za pomocą VOC

6. PODSUMOWANIE 

7. WNIOSKI 

LITERATURA

SUMMARY

A series of experiments was carried out with the aim to answer the question whether there were differences in the formation by fungi of volatile metabolites in the case of infestation of untreated timber in comparison with the timber impregnated below the limiting fungicidal value. The following three fungicides were used to treat experimental timber below the fungicidal value: 3-iodo-2-propynyl-butylcarbamate – IPBC, didecyl dimethyl ammonium chloride – DDAC, 1-[[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl]methyl]-1,2,4-triazole; 1H-1,2,4-triazole – propiconazol. The technique of solid phase microextraction (SPME) was employed to isolate volatile metabolites emitted by test fungi, whereas a gas chromatograph coupled with a mass spectrometer were used to identify them. The paper presents quantities and profiles of chemical compounds secreted by test fungi at the following time intervals: 4, 7, 10 and 14 days. During the performed experiments on the dynamics of the formation of volatile metabolites, attention was focused on compounds that can exert a potentially allergic action on homoeothermic organisms and can also be utilised as indicators of the presence of fungi in the environment.
The following four fungi were selected for the experiments: Aspergillus niger van Tieghem, Trichoderma viride Persoon ex S.F. Gray aggr. Gloeophyllum trabeum (Persoon ex Fries) Murrill, Coniophora puteana (Schumacher ex Fries) Karsten which were cultured on Scots pine (Pinus sylvestris) timber.
On the basis of the performed mycological tests, the following results were obtained for individual fungi. A. niger infesting Scots pine wood emitted a number of compounds, among others, ketones containing up to 8 carbon compounds in one molecule (pentan-2-one, heptan-2-one, 1-octen-3-one, octan-3-one, octan-2-one). Another numerous group of volatile metabolites comprised alcohols (heptan-1-ol, oct-1-en-3-ol, 3-octen-2-ol and octan-2-ol), monoterpenes: (α-pinene, sabinene, limonene, D-pinocamphone, dichydrocarvone) as well as terpene derivatives (myrtenol and isopulegol acetate), diterpenes, (18-nor-4(19),7,15-iso-pimaratriene, rimuene, manool, 13-isopimaradiene, phyllocladene, abietatriene), esters (heptyl hexanoate, 3-methylotridecane acid methyl ester, dodecanoic acid 1-methyl ester, isopropyl myristate), furan derivatives (furfural and 2-noctylofuran). In the case of the synthesis of the majority of volatile metabolites emitted by A. niger degrading the control timber and the one treated with fungicides, the highest level was usually observed on the fourth day of culturing. The most conspicuous changes in concentration levels were recorded between days 4 and 7 of the test duration and they were noticeable in the case of nearly all volatile organic compounds (VOC) with the exception of oct-1-en-3-ol. Furthermore, it was found that the biosynthesis of such compounds as oktan-2-one, heptan-2-one, pentan-2-one, oct-1-en-3-ol, 3-octen-2-ol could be stimulated by the content of fungicides below the limiting fungicidal value in wood. However, it was impossible to determine precise relations between the specific fungicide, its concentration and emissions of the above-mentioned compounds by A. niger.
The successive microfungus that was investigated in the presented experiments was T. viride. In the course of experiments, 47 compounds were extracted from the air above the surface of the mycelium. The most numerous class of chemical compounds emitted by T. viride turned out to be alcohols followed by ketones then: monoterpenes, sequiterpenes, esters and diterpenes, unidentified compounds of 204 molecular weight corresponding to the fraction of sequiterpenes hydrocarbons and of 272 molecular weight attributed to diterpenes. During culturing, the highest concentrations were recorded for Eight-carbon compounds such as: octan-2-ol, octan-3-ol, octan-3-one, octan-2-one, oct-1-en-3-ol. It was possible to observe, similar as for A. niger, regularity of the dynamics of VOC emissions by T. viride developing on the control timber. In both cases, the highest concentrations of volatile metabolites were recorded on days 4 and 7 of measurements. The smallest quantities of volatile metabolites, with the exception of oct-1-en-3-ol, were determined on the last day of experiments. Analysing the VOC emission dynamics by T. viride growing on the wood treated below the limiting fungicidal value, increased emission of such compounds as: norinone, pentan-2-one, octan-2-one was observed. On the basis of the performed investigations, it was not possible to propose pathways of biochemical changes taking place in the course of the development of the test microfungi on timber containing fungicides below limiting fungicidal value.
The next fungus employed in these studies was C. puteana. On the basis of model investigations of the fungus degrading Scots pine wood, it was possible to identify 48 compounds classified as: alcohols, esters, carboxylic acids, terpenes and diterpenes. Among dominant volatile metabolites were: γ-cadinene, octan-3-one, oct-2-en-1-ol, oct-3-en-2-ol, hexanol, myristic acid and palmitic acid. Determining the synthetic capability of C. puteana in relation to the time of culturing, it was found that the most diverse, with respect to quality and quantity, chromatographic profile occurred on days 4 and 7. The only exception was the oct-1-en-3-ol alcohol whose synthesis increased from the level of day 4 up to day 10 and on the last day of measurements it was comparable with day 7. In the case of VOC emissions of C. puteana degrading wood treated with fungicides below the limiting fungicidal value, inhibition of synthesis was observed for the majority of volatile metabolites. Only for one variant – octen-2-ol – stimulation was recorded on mycelium samples on day four of culturing.
G. trabeum developing on Scots pine wood emitted 50 volatile metabolites which comprised the following classes of chemical compounds: alcohols, aldehydes, esters, ketones, aromatic compounds but, first and foremost, monoterpenes and sequiterpenes hydrocarbons as well as various terpene derivatives. The highest concentrations were found for sequiterpenes: citronellol, α-neoclavene, β-elmene, unidentified sequiterpene as well as an unidentified compound and eight-carbon alcohols: oct-1-en-3-ol and 2-okten-1-ol. The most numerous group of volatile metabolites isolated from the abovesurface phase included esters. The total of 14 esters was successfully identified. In the case of the majority of volatile metabolites secreted by G. trabeum, it was found that the highest concentrations of these compounds occurred on the fourth day and were found in the air above the surface of the mycelium. The exceptions included: oct-1-en-1-ol and limonen. Examining variants with wood containing fungicides below the limiting fungicidal value, a phenomenon of synthesis inhibition was observed for hexan-1-ol and oct-1-en-3-ol by the test fungus in comparison with the development of mycelium on the control samples. The only exception was the case when the mycelium settled wood treated with DDAC because no significant differences were observed in the emission of oct-1-en-3-ol on the last day of the mycological test. For the majority of variants, it was impossible to identify the phenomenon of inhibition or stimulation depending on the concentration and kind of fungicide in timber. In most variants, terpenes (limonene, α-neoklovene, and β-elemene) were synthesized by G. trabeum at higher concentrations in comparison with control samples.
It was concluded, on the basis of the performed mycological tests that eight-carbon alcohols and ketones can serve as indicators of the presence of A. niger, T. viride, C. puteana and G. trabeum in facilities in cases when the fungi develop in wood which does not contain fungicides. In addition, it can be said that individual species of fungi inoculated on samples containing fungicides (IPBC, DDAC and propoconazol) below fungicidal values release a wide range, with regard to their quantity and quality, of volatile metabolites in comparison with the variants which did not contain fungicides. For this reason, eight-carbon alcohols and ketones cannot be used as markers of fungal presence in different facilities.