Black fungal extremes - CBS

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Sat o w e t a l.Assimilation of mineral oil and n-hexadecaneThe readings of 80 % hydrocarbon concentration were notconsidered because at this percentage an interference on theabsorbance occurred. Growth curves of the strains in cultureconditions A, B and C (values with blank discounted) are shown(Figs 1–3).Strains not belonging to the black yeast group: Rhodotorulagraminis (CBS 2826) and Penicillium chrysogenum (CBS 776.95)showed minimum growth in culture conditions A, B and C (Figs1–3). Similar growth was observed for the black yeast strain CBS122275. Ability to use hydrocarbons as sole source of carbon andenergy seemed to be absent for these strains.Cladophialophora sp. CBS 122255 showed growth in all cultureconditions, however its growth was higher in culture condition A,without hydrocarbon (Fig. 1). Better growth with hydrocarbonas sole source of carbon and energy was observed in strainsFig. 1. Culture condition A: growth curves of studied strains in mineral medium, YNB (continuous line), or in physiological salt solution, NaCl (line and dots).Fig. 2. Culture condition B: growth curves of studied strains in mineral medium (YNB) + 20 % mineral oil (continuous line), and in physiological salt solution (NaCl) + 20 %mineral oil (line and dots).160
Oil f l o tat i o n is o l at i o n o f b l a c k y e a s t sFig. 3. Culture condition C: growth curves of studied strains in mineral medium (YNB) + 20 % n-hexadecane (continuous line), and in physiological salt solution (NaCl) + 20 %n-hexadecane (line and dots).CBS 122253, CBS 122257, CBS 122258 and CBS 118157. Thedifference between the growth rates in culture conditions B andC were small: strain CBS 122253 and CBS 122258 showed thesame rates for both culture conditions; higher growth rates withmineral oil was observed for strain CBS 122257; and strain CBS118157 seemed to be stimulated by adding n-hexadecane (Figs2–3). Strain dH 18473 had similar growth in culture conditions A, Band C (Figs 1–3).OligotrophismWhen growth was tested in physiological salt solution, severalblack yeasts and relatives (CBS 122258; CBS 122257; CBS122253; CBS 118157) were able to grow for few ds. However,growth rates were lower than in assimilation tests as presumeddue to the poor concentration of nutrients. Stationary phase wasmostly reached after 36−68 h. Cladophialophora minourae (CBS122275) and Cladophialophora sp. (dH 18473) showed the samepattern as reference strains (Rhodotorula graminis and Penicilliumchrysogenum): they all did not have significant growth in NaCl(Figs 1–3). Surprisingly some growth curves in mineral oil andn-hexadecane 20 % from oligotrophism tests were higher thanobserved in assimilation tests. Strain CBS 122258 and CBS 118157(both Exophiala xenobiotica strains) continued growth at a minimallevel. Addition of 20 % mineral oil slightly stimulated growth of blackyeast-like fungi in that stationary phase was reached somewhatearlier (Fig. 2). Addition of 20 % n-hexadecane stimulated growthin Exophiala strains that performed best in previous tests; clearexponential and stationary phases were distinguishable (Fig. 3).Strains CBS 122255 and dH 18473 also did not show significantgrowth in NaCl. Strains CBS 122257 and CBS 122253 showedability to grow in media poor in nutrients; growth levels werenot so high as for Exophiala strains but were higher than inCladophialophora sp.www.studiesinmycology.orgDISCUSSIONPrevious studies on black yeasts isolation using the oil flotationtechnique reported that soil was poor in black yeast (Iwatsu et al.,1981; Marques et al., 2006; Vicente et al., 2001), but in the presentarticle using polluted soil, a large number of isolates was obtained.In these researches few black yeast strains per sample wererecovered: Iwatsu et al. (1981) isolated 83 dematiaceous strainsfrom 177 samples; Vicente (2000) obtained 81 strains from 540samples and Marques et al. (2006) 9 isolates from 68 samples. Inthis study 107 strains were isolated from 3 samples. Hence it maybe concluded that hydrocarbons present in the soil might favor thegrowth of the black yeasts and function as an enrichment factor, assupposed earlier (Prenafeta-Boldú et al. 2002, 2006, Sterflinger &Prillinger 2001) or inhibited growth of competing species. Judgingfrom the number of strains recovered, and the number of strainsfor which no match was obtained with any known species, landfarming soil was considered an interesting substrate for recoveringthis group of fungi. Studies on the ecology, phylogeny andbioremediation seem to be promising. Previous studies in this areaobtained strains able to degrade aromatic hydrocarbons such asfungi from genera Fusarium, Penicillium, Trichoderma, Aspergillus(Kataoka 2001, Satow 2005) and some fungi with black colonies,however, molecular identification was not done to confirm if theybelonged to black yeast-like fungi (Conceição et al. 2005).The recovery of Exophiala xenobiotica from land farming soilconfirms that particularly those members of Chaetothyriales showinga preference for habitats rich in monoaromatic hydrocarbons andalkanes are recovered (de Hoog et al. 2006, Sterflinger & Prillinger2001, Woertz et al. 2001). Given their potential as opportunists orhuman pathogens (e.g., Prenafeta-Boldú et al. 2006, Zeng et al.2007) the results of this study may contribute for the biosafety rulesestablished for the employees of the refinery. On the other hand,black yeasts and relatives could be also developed as agents forbioremediation for industrial purposes, such as treatment of volatile161
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Studies in Mycology 61 (2008)Black
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Studies in MycologyThe Studies in M
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PREFACEThe terms "black fungi" or "
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available online at www.studiesinmy
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Dr o u g h t m e e t s a c i dTable
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Dr o u g h t m e e t s a c i d2003/
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Dr o u g h t m e e t s a c i din Pe
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Dr o u g h t m e e t s a c i d99100
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Dr o u g h t m e e t s a c i dAnamo
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Dr o u g h t m e e t s a c i dFig.
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Dr o u g h t m e e t s a c i dconve
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Dr o u g h t m e e t s a c i dal. 2
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Za l a r e t a l.Table 1. List of a
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Za l a r e t a l.Table 1. (Continue
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Za l a r e t a l.Fig. 2. Consensus
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Za l a r e t a l.Fig. 4. Macromorph
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Za l a r e t a l.Fig. 5. Aureobasid
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Za l a r e t a l.Fig. 7. Aureobasid
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Za l a r e t a l.the type species o
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Za l a r e t a l.Vishniac HS, Onofr
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Yu r l o va e t a l.Table 1. Strain
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Yu r l o va e t a l.112ОС2Fig. 1.
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Yu r l o va e t a l.RESULTSFourteen
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Yu r l o va e t a l.100908070605040
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Yu r l o va e t a l.AcetateMelanin(
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Va u p o t i č e t a l.HMG-CoA red
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Pl e m e n i ta s e t a l.entire en
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Pl e m e n i ta s e t a l.glycerol
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Pl e m e n i ta s e t a l.Fig. 1. T
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Pl e m e n i ta s e t a l.diversity
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Ti n e a n i g r aand Polynesia (Ue
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On o f r i e t a l.Table 1. Test pa
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On o f r i e t a l.Analyses of resp
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On o f r i e t a l.Fig. 6. Percenta
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Page 124 and 125: De n g e t a l.Table 1. Strains use
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Page 192 and 193: homogentisate 165, 171, 173Hortaea
Page 194 and 195: CBS Biodiversity SeriesNo. 6: Alter
Page 196 and 197: Studies in Mycology (ISSN 0166-0616
Page 198: Satow MM, Attili-Angelis D, Hoog GS
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