KRZYSZTOF ULFIG, MACIEJ TERAKOWSKI, GRAŻYNA PŁAZA
F IR S T IS O L A T IO N O F M IC R O SP O R U M R A C E M O S U M B O R E L L I IN P O L A N D Institute for Ecology of Industrial Areas,
40-832 Katowice, Kossutha 6 Str., Poland Head: doc. dr hab. E. Marchwińska
During a study o f keratinolytic fungi in the ash heap ground, considerably influenced by the municipal landfill site, a rare potentially pathogenic dermato phyte, Microsporum racemosum Borelli, was recognized. This is the first report on its isolation in Poland. Morphological and ecological features o f the fungus are discussed.
INTRODUCTION
D u rin g a study o f keratinolytic fungi (K F) in soils contam inated by m unicipal landfill sites [17], a ra re pathogenic derm atophyte, M icrosporum racem osum Borelli, was isolated from th e ash heap ground. In this article, we p resen t its m acro- and m icroscopic characteristics as well as discuss possible ecological factors affecting the incidence o f th e fungus in th e ground.
MATERIAL AND METHODS
The ash heap investigated is situated in Sosnowiec at a distance of ca. 100 m from the municipal landfill site. On this heap, coal combustion ash from the coal mine „Kazimierz-Juli- usz” was dumping for a period of ca. 100 years. The dumping was ceased in 1990. Up-till-now, the ash heap surface (ca. 1.5 ha) has not been treated in any way. Actually, it is covered with the black „coal” ground (loose sand in the ground classification by the Polish Standard BN-78/9180-11), on which only few plants (lichens, mosses, herbs, grasses and shrubs) are possible to be found. Ash heap's material is also used as inert material for municipal waste disposal.
Ash heap ground was sampled 4 times (April, October 1993; May, September 1994). Altogether, 7 locations on ash heap's surface were investigated. The way of sampling is de scribed by Ulfig [17]. The samples were examined for selected physico-chemical parameters (Tab. 1). Full physico-chemical characteristics of the ash heap material were presented by Terakowski [15]. The human hair baiting technique [19] was used for KF recognition in the samples. Isolated fungi were identified according to selected keys and monographs [3, 8, 13].
RESULTS
O u t o f 90 Petri dishes set up, 63 (70% ) w ere positive for KF. A lto g eth er, 158 fungal ap p earan ces belonging to 18 species w ere observed. L index (n u m b er o f ap p earan ces divided by the n u m b er o f Petri dishes set up) was 1,76. A phanoascus durus w ith its an am o rp h Chrysosporium sp. and Ch. europae w ere the prevailing species in th e ash
314 К. Ulfig i in. N r 3 T a b l e I . Physico-chemical characteristics of the ash heap ground
dw - dry weight of ground
heap ground (Fig. 1). Microsporum racemosum w ere found to grow on 4 Petri dishes. T h e species ap p e ared in A pril 1993. Two strains w ere isolated and intro d u ced to th e IE IA (In stitu te fo r Ecology o f Industrial A reas) collection.
M i c r o s p o r u m r a c e m o s u m B o r e l l i
O n Sabouraud glucose agar (SG A ), colony growing m oderately rapidly, w ith a m ean daily grow th ra te o f 4 mm in the dark at 28°C, flat, with a folded and pinkish um bo, downy through floccose to granulose o r powdery, initially colourless, later becom ing d ark cream to pink; m argin defined, irregular, fim briate; reverse initially yellowish but slowly becom ing brow nish, brown to pink-violet; pigm ent not diffusing into th e agar. T he pigm ent also p roduced on p o tato dextrose agar (P D A ) and, though only brow n, on phyton yeast extract agar (PY E ). N o pigm ent observed on m alt extract agar (M E A ) and Sabouraud 1:10 + salts ( Takashio agar).
H yphae hyaline, septate, branched, thin- and sm ooth-w alled, 1.8—4.5 ц wide. No racq u et and spiral hyphae observed. M icroconidia extrem ely num erous, disposed in clusters, usually arises from the conidiogenous hyphae at right angles, hyaline, thin- and sm ooth-w alled, stick- and cartridge-shaped, with tru n cate basal ends, 1-, rarely
Fig. 1. Occurence of keratinolytic fungi in the ash heap ground in Sosnowiec
Abbreviations: TAJ - Trichophyton ajelloi ACUR - Arihrodenna curreyi CACUR - Chryso- sporium an. Arthroderma curreyi CTROP - Chrysosporium tropicum MFLA - Malbranchea flava ARET - Aphanoascus reticulisporus AFUL - Aphanoascus fulvescens CHRYS - Chrysosporium an. Aphanoascus reticulisporus /fulvescens CKER - Chrysosporium keratinophilum MGYPS - Microsporum gypseum complex MVEL - Myceliophthora vellerea CEUR - Chrysosporium europae CIND - Chrysosporium indicum ADUR - Aphanoascus durus CADUR - Chrysosporium an. Aphanoascus durus CPAN - Chrysosporium pannicola TTER - Trichophyton terrestre complex AQ - Arthroderma quadrifidum CMEF - Chrysosporium mephiticum GYMN - Gymnoascus sp.
MRAC - Microsporum racemosum CACUN - Chrysosporium an. Arthroderma cuniculi 2-celled, 2.7-9.7 x 1.3-3.3 p m . M acroconidia less num erous, growing on sim ple, unbran ch ed conidiophores, hyaline, m ultiseptate, fusiform , elliptical and cigar-shaped, relatively thin-w alled (0.8-1.6 p m) , generally sm ooth, som e o f them verruculose o r verrucose, on SG A regularly shaped and (1 -6 ) - celled, on S abouraud 1:10 + salts (1-12, usually 7 -9 ) - celled, often curved or even branched, 24.4-71.0 x 8.0-12.6 p m (Fig. 2, 3). N o chlam ydospores observed. N o ascam ata observed on the h air bait and w hen crossed the strains isolated. O ne of the strains (IE IA 672A) was sent to the M icrofungus Collection and H erbarium , University o f A lberta, E dm o n to n , C an ad a (U A M H ).
T he fungus does not grow at 37°C but displays slow grow th at 7°C, with a m ean daily grow th rate o f 0.37 mm. Strongly keratinolytic, producing p en etratin g organs in the hair.
316 К. Ulfig i in. N r 3 M a t e r i a l e x a m i n e d .
Living single-spore strains (obtained by the dilution m ethod): IE IA 673A and IE IA 674B, ash heap ground, Sosnowiec, Poland.
DISCUSSION
M. racem osum was first described in 1965 in V enezuela w here was isolated from th e healthy h air o f a forest ra t [2]. Subsequently, the species was recovered in A m azonias soil, and also from forest soil and anim al h air (squirrel, badger) in R o m an ia [1, 12]. In 1982, soil isolates o f M. racemosum w ere also rep o rte d in Czechoslovakia [2]. Pathogenic p ro p erties o f the fungus w ere d em o n strated bo th experim entally and naturally [1, 2, 4, 5, 11, 12]. Its perfect state, Nannizzia racemosa R ush-M unro, Sm ith et B orelli, was described by R ush-M unro et al. [12]. In 1986 [21], a new com bination, Arthroderm a racem osum , was proposed.
M racem osum is very close to the m em bers o f the M. gypseum -fulvum group. T h e first is characterized by the ab u n d an t stick- and cartridge-shaped m icroconidia disposed in clusters w hereas in th e M. gypseum-fulvum group the m icroconidia are usually clavate and never the prevailing m icrom orphological elem ents [14]. B ecause o f th e abun d an ce o f m icroconidia, th e m icrom orphology o f M. racem osum , at first glance, is m ore characteristic fo r Trichophyton than Microsporum species. Subsequently, exam ination o f m acroconidia reveals th a t in M. gypseum-fulvum group they are typically verruculose to verrucose while in M. racem osum a considerable p a rt o f th em is sm ooth th a t also resem bles Trichophyton. A dditionally, the n u m b er of septa, generally uniform in M. gypseum (up to 6), is usually higher in M. racem osum (up to 12). Finally, the pro d u ctio n o f the characteristic pink-violet pigm ent by M. racem osum is o f p articu lar taxonom ic significance. T he pigm ent appears a fter 3 w eeks o f incubation b u t only on S G A and PD A .
Although K F in soils and o th e r habitats in P oland have been a subject o f extensive investigation fo r m ore th an thirty years, th ere is no referen ce to M. racem osum isolates
in available literatu re [6, 7, 9, 10, 16]. This is, th erefo re, the first re p o rt on its isolation in o u r country. It is also to note th a t a sim ilar fungus was previously isolated from sewage sludge but m isidentified as a m em ber o f the M. gypseum -fulvum group [18]. T his d ata indicates th at M. racemosum is a rare but widely distributed pathogenic fungus.
T h e h ab itat o f the ash heap ground is of particular ecological interest. This ground is h etero g en o u s as regards physico-chemical features. In general, it is characterized by th e high ignition losses and organic carbon (T jurin), derived from th e partly com busted coal, low salt concentration, and pH close to neutral. A p a rt from the Z n co n ten t, the con cen tratio n of the o th e r heavy m etals w ere low. T he untoxic c h aracter o f th e ground was show ed by the w atercress sprout test [15]. U n d e r these ra th e r favourable conditions, lower and higher plants as well as m icro-organism s w ere poorly re p resen te d [17]. T his was probably caused by the low n u trien t co n centration and the deficiency o f available carbon sources in the surface layer o f the ground. In addition, the ground ten d ed to get dried rapidly during the intensive insolation and dry w hether.
In contrast, Kf w ere found to occur relatively aboundantly in the ash heap ground. M. racem osum was strictly associated with this habitat. This finding is ra th e r opposed to previous observations indicating the association o f M. racem osum chiefly with forest soils and anim al hair. T h e presence and good growth of the species in the ash heap ground can be explained in the following way: T he fresh coal com bustion ash a m a terial devoid o f living organism s and keratin rem ains need ed for the grow th o f KF. T he only im p o rtan t source o f keratin and K F in the surroundings o f the ash heap is the m unicipal landfill site. This object considerably influenced, via the air, the ash ground m ykoflora. Som e KF, including M. racemosum, found favourable conditions to survive in this habitat. H ow ever, this derm atophyte did not occur in m unicipal landfill site's ground. This can be explained by the unfavourable ecological conditions of the ground and the possible restricted abilities o f the fungus to com pete with o th e r KF on hair bait.
318 К. Ulfig i in. N r 3 M unicipal solid w astes and th e habitats influenced by m unicipal landfill sites can be real sources o f pathogenic fungi [17]. Isolation o f pathogenic d erm atophytes from this environm ent can n o t be ignored from an epidem iological p oint o f view.
K . U l f i g , M . T e r a k o w s k i , G . P ł a z a
PIERWSZA IZOLACJA MICROSPORUM RACEMOSUM BORELLI S t r e s z c z e n i e
W toku badań grzybów keratynolitycznych w glebach zanieczyszczonych przez wysypiska odpadów komunalnych z hałdy popiołów kopalnianych wyizolowano rzadki gatunek chorobot wórczego dermatofita - Microsporum racemosum Borelli. W niniejszym artykule przedstawiono makro- i mikroskopowe właściwości tego gatunku, jak również przedyskutowano potencjalne ekologiczne czynniki wpływające na występowanie grzyba w gruncie badanej hałdy.
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