Mycologia, 97(1), 2005, pp. 33–44.

2005 by The Mycological Society of America, Lawrence, KS 66044-8897

Isolation and identification of fungal communities in compost and vermicompost
Antonella Anastasi Giovanna Cristina Varese1 Valeria Filipello Marchisio
Department of Plant Biology, Viale Mattioli 25, 10125, Turin, Italy

Abstract: This research illustrates the qualitative and quantitative composition of the mycoflora of both a green compost (thermophilically produced from plant debris) and a vermicompost (mesophilically produced by the action of earthworms on plant and animal wastes after thermophilic preconditioning). Fungi were isolated using three media (PDA, CMC, PDA plus cycloheximide), incubated at three temperatures (24, 37 and 45 C). Substantial qualiquantitative differences in the species composition of the two composts were observed. The total fungal load was up to 8.2 105 CFU/g dwt in compost and 4.0 105 CFU/g dwt in vermicompost. A total of 194 entities were isolated: 118 from green compost, 142 from vermicompost; 66 were common to both. Structural characterization of this kind is necessary to determine the most appropriate application of a compost and its hygienic quality. Key words: compost, compost hygiene, compost quality, earthworms, fungi

INTRODUCTION

Composting is the biological conversion of solid organic waste into usable end products such as fertilizers, substrates for mushroom production and biogas. Moreover, their high organic matter content and biological activity make composts effective in a variety of applications, including erosion control, revegetation, biofiltration and bioremediation (Alexander 1999). The active component involved in the biodegradation and conversion processes during composting is the resident microbial community, among which fungi play a very important role. The biomass ratio of fungi to prokaryotes in compost is about 2:1 (Sparling et al 1982, Wiegant 1992). In addition, fungi use
Accepted for publication 18 Aug 2004. 1 Corresponding author. Email: cristina.varese@unito.it

many carbon sources, mainly lignocellulosic polymers and can survive in extreme conditions. They mainly are responsible for compost maturation (Miller 1996). A better understanding of fungal diversity in compost may prove crucial in predicting its best application. Fungi affect soil fertility, suppress plant diseases and promote mushroom growth (Straatsma and Samson 1993). They also degrade complex polymers such as polyaromatic compounds or plastics and are being increasingly applied to bioremediate soils contaminated with a wide range of pollutants (Kastner and Mahro 1996, Eggen and Sveum 1999, Minussi et al 2001). Monitoring fungal diversity is essential to detect fungi hazardous to humans, animals and plants and to optimize compost quality standards (Summerbell et al 1994). Much information exists about the succession of fungi, mainly thermotolerant and thermophilic fungi, in conventional two-phase thermogenic composting (Straatsma et al 1994, Ross and Harris 1983, Fermor et al 1979, Chang and Hudson 1967). These data refer mainly to mushroom compost, straw compost or experimental compost obtained by environmentally controlled and standardized processes. However, industrial composting uses a variety of procedures and raw materials (Beffa et al 1998) and hence results in very different end products. In contrast, very little is known about fungal communities in mesophilic processes such as vermicomposting, an alternative technology increasingly used in many countries, including Italy (Beffa et al 1998, Masciandaro et al 2000). Earthworms stabilize organic residues and reduce pathogenic bacteria and other human pathogens (Eastman et al 2001) and also can greatly affect fungal communities. They select fungal species by influencing spore germination and creating microsites favorable or unfavorable to fungus development (Brown 1995, Tiunov and Scheu 2000). The few studies on these mechanisms have provided partly contradictory data and stressed the importance of monitoring the hygienic aspects of this mesophilic process in fungal communities (Beffa et al 1998). In brief, since composting methods and different source materials are associated with differences in the composition of a fungal community, monitoring of the resident fungal population in a compost is 33

After determination of their genera (Domsch et al 1980.9%). the moisture content 40%.34 MYCOLOGIA clamp connections or positive to the reaction with Diazonium Blue B salts (DBB). The Mann-Whitney test (StatView 1988) was run to assess the significance (P 0.6% of the dry matter and the C/N ratio about 25. further dilutions were made in NaCl (0. In C. the two population structures were analyzed with the rank-abundance plot (Biodiversity PRO 1997). During this mesophilic phase the temperature never exceeded 25–30 C. 50 m long. Both composts were stored in polypropylene bags 1–3 mo at 10 C before being sold. a significant reduction in CFU/g dwt was induced by higher incubation temperatures and the addition of cycloheximide.3 10 105 CFU/g dwt in VC. needed to determine its quality and field of application (Peters et al 2000). Compost (C) was produced in an outdoor pile (3 m wide. Ten approximately 1 kg samples per compost (C1–10. The pH of the final product was 7. Filipello Marchisio et al 1996).0 from 5. the Berger-Parker index from the formula d Nmax/N (where Nmax is the number of individuals in the most abundant species). d and D decrease. Hanlin 1990. species and genera load) and between all treatments (three media and three incubation temperatures) in the composts. depending on media or incubation temperature (TABLE I). poultry and various zoo animals) and 30% plant debris from various sources. Employment of the CMC and CX media.05) of the differences of each index between the two composts. humic organic carbon (humic acid fulvic acid) 3. The nonparametric Mann-Whitney test for independent groups (StatView 1988) was run to assess the significance (P 0.5 m high) composed of 70% dung (from cows. In VC. VC1–10). The maximum temperature during the composting was 60 C. and incubation at 37 C and 45 C allowed . MATERIALS AND METHODS The following physical dimensions. Diversity indexes based on species richness (Margalef index) and species relative abundance (BergerParker. A total of 194 fungal entities were identified from the two composts. SM with RESULTS The total fungal load was high in both composts: from 5. We therefore used these indexes in their reciprocal form 1/d and 1/D (Magurran 1988). temperatures and moisture measurements were taken outdoors and are approximations.0 104 to 8. unless denoted otherwise.2.5%. Plates were incubated at 24 C. during which the piles were turned periodically by machine. Multivariate analysis (Detrended Correspondence Analysis-DCA) was used to evaluate quali-quantitative differences in the composition of the mycofloras of the two composts and between the 10 samples of each compost (CANOCO 1998). the Simpson index from the formula D {[ni(ni 1)]/[N(N 1)]}. 0. Shannon. three of carboxy-methyl cellulose agar (CMC) and three of PDA supplemented with cycloheximide (CX) to retard the growth of all fungi. allow isolation of slow-growing colonies and focus on fungi of medical interest (Airaudi and Filipello Marchisio 1996. Moreover. higher temperatures.05) of the differences between the two composts (total load. von Arx 1981. Simpson indexes) were applied to assess biodiversity (Biodiversity PRO 1997). and 4 to 4. All statistics were obtained from the highest load of each species in the 9 treatments of each sample. The pH of the final product was 7. earthworms (Lumbricus rubellus Hoffmeister) were added (50 103 worms per m3 organic matter) and the pile was turned periodically by machine. This work focuses on the species composition and load of the mycoflora of two mature composts marketed by an Italian firm: a compost currently used as a bioactivator in landfills and a vermicompost mainly applied in agriculture. of which 118 came from C and 142 from VC. 37 C and 45 C to isolate mesophilic and thermotolerant/ thermophilic fungi with the result that 33 replicates were made for each sample. during which the temperature rose to 60 C. Kiffer and Morelet 1997). sclerotia or vesicles. humic organic carbon (humic acid fulvic acid) 7. they were transferred to the media recommended by the authors of selected genus monographs for species identification.9. According to Magurran (1988).6% of the dry matter and the C/N ratio 15.2 105 CFU/g dwt in C. were examined according to the guidelines proposed by the Piedmont Region (Trombetta et al 1998). Sterile mycelia (SM) were classified according to their hyphal pigments and their production of chlamydospores.5 m high) from plant debris from various sources by a conventional thermophilic process that lasted ca 6 mo. After preconditioning for several days. according to Summerbell (1985). The greatest number of species were isolated from both composts on PDA incubated at 24 C. 1. The final dilution (1: 20 000) was plated (1 ml per plate) on 11 replicates: five of potato-dextrose agar (PDA). Only 66 were common to both composts (TABLE II). Vermicompost (VC) was produced in an outdoor pile (3 m wide. S is the number of fungal entities and N is the total number of individuals). except for CMC at 45 C and CX at all temperatures (TABLE I). As diversity increases. The number of colony forming units per g of dry weight (CFU/g dwt) was calculated both for the total mycoflora and for each species or morphotype. the Margalef index was calculated from the formula DMg (S 1)/ln N (here and throughout. were classified as basidiomycetes. cycloheximide and carboxy-methyl cellulose all reduced CFU/g dwt values. The load values in C always were higher. Fungi were identified conventionally according to their macroscopic and microscopic features. A 10 g portion of each sample was suspended in 90 ml Na4P2O7·10 H2O to disperse organic colloids. however. the moisture content 38. the Shannon index from the formula H Spi(lnpi) (where pi is the proportion of individuals found in the ith species). The culture and/or incubation conditions produced different load values within the same compost. 50 m long.

6·105 1. Both loads were composed mainly of thermotolerant A.3. the isolation of a good number of species that otherwise would have been missed: 24 from C and 30 from VC (TABLE I). and many species solely were present in C or in VC (TABLE II). ellipticus. Chrysosporium and Scopulariopsis species prevailed in VC (respectively 1.0·104 a* a* b* c* ac* bc c c bc No.1·104 2. of fungal entities 70 42 43 32 17 9 25 5 3 (31) (6) (7) (5) (3) (0) (3) (0) (0) 4. Both composts were dominated by mitosporic fungi (including the ascomycetes in their anamorphic state) (TABLE IV).1 104 in VC versus 1. There are three zones along the 1 axis: zone I containing most of the C samples (C4–10) and the entities found only or preponderant in C. 45 ascomycetes. 37 C.0·104 3.6·105 1. the thermotolerant fungus Scedosporium state of Pseudallescheria boydii displayed a significantly greater load (P 0. 2). Eurotium chevalieri (mainly present in VC6).6·105 2.0·104 4.8 105 CFU/g dwt in C and 1. 1).5·104 5.8·105 2. and 3. the greater number of species corresponds to higher biodiversity index values (TABLE III). though P. fumigatus (TABLE II). mainly in C4 (FIG. mainly .1 104 in C versus 2.0·105 2. Other species described as thermotolerant or thermophilic (Domsch et al 1980) were isolated at 37 and 45 C from both composts with no significant load differences. 14 SM morphotypes and three basidiomycete morphotypes (TABLE II). roseopurpureum (P 0.2 105 CFU/g dwt in C.5·105 9.2·105 8. This difference is not signif- icant. zone III containing most of the VC samples (VC1. Malbranchea cinnamomea.1·104 a* a* b* a* ab* bc d d c No.7 104 CFU/ g dwt in C). zone II containing C and VC samples (C1–3 e VC3.5·105 1.013) and P. They included Aspergillus fumigatus var.0 105 CFU/g dwt in VC and 1. Of the most abundant species in both composts.0012) in C (7. or found only in C or in VC. The total load of Penicillium was 3.4·105 1.2 103 CFU/g dwt) were present exclusively in C.6·104 2. namely the Scedosporium state of Pseudallescheria boydii and Aspergillus fumigatus. and Trichoderma species (8. particularly owing to the presence of Corynascus sepedonium (mainly present in VC3). There were no significant differences between composts in the quantitative composition of the two sets of Cladosporium and Acremonium species (about 5. CMC. Paecilomyces variotii and Thermomyces lanuginosus.9·104 5. 2).9·104 4. The genera with the highest load and number of species in both composts were Penicillium and Aspergillus. fumigatus var. CX) incubated at 24 C.0·105 2. 2.4·105 5.03) showed prevalence in C and associated with C8 and C5 respectively (FIG.3·104 VC 8. aurantiogriseum (P 0. which demonstrates the quantitative domination of two species.5·105 9. Mann-Whitney test) among the load of the same compost obtained in different culture condition and/or incubation temperatures and * indicates significant differences between C and VC in the same culture conditions. 2). flavus (mainly present in VC2. The Aspergillus load was not significantly different: 1.1 103 CFU/g dwt in VC).1·105 1. Mean fungal load (CFU/g dwt SE) and number of fungal entities isolated in compost (C) and vermicompost (VC) on 3 media (PDA.1·105 3. 4–7) and the entities found only or preponderant in VC.8·104 6. In VC.0 104 and 1. The DCA scatterplot (FIG. whereas Fusarium species prevailed in C (3.3 105 CFU/g dwt) and was associated with it in the DCA and included in zone I (FIG. and Talaromyces flavus var. but present in smaller quantities and thus regarded as occasionals (FIG.105 CFU/g dwt in VC. 8–10) and the entities equally distributed between C and VC.2 104 in VC versus 0 CFU/g dwt in C. 2). 2). 2) shows the distribution of the samples and the 194 fungal entities.5·104 9.8·105 6. FIG.1·104 4. 15 zygomycetes. The lower evenness of C is illustrated in the rank abundance plot (FIG. Number of entities isolated from C only and from VC only in brackets C PDA 24 C PDA 37 C PDA 45 C CMC 24 C CMC 37 C CMC 45 C CX 24 C CX 37 C CX 45 C 8. 45 C.10) (TABLE II.0·104 1.ANASTASI ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 35 TABLE I. aurantiogriseum var.3·105 1.0 104 CFU/g dwt respectively in both composts).2·105 1.7·104 2.3·105 7. 2).4·105 4. of fungal entities 98 52 37 30 19 15 30 19 6 (46) (11) (3) (4) (4) (1) (2) (4) (1) Different letters indicate significant differences (P 0. which displayed high loads in all the samples and was located in zone II (FIG. The load of zygomycetes was similar in C and VC with greater species diversity in C (TABLE IV).05. The number of species and the load of ascomycetes were higher in VC (TABLE IV). The 194 fungal entities comprised 117 mitosporic fungi.9·104 1.0·104 2. Absidia corymbifera alone was isolated from C only.

1·104 1.0·102 3. 1 Acremonium sp.0·103 9.0·103 2.4·103 1.9·103 1.8·102 1.3 102 — — 7. Emmons) von Arx Corynascus sepedonium (C.3·103 6.3·102 1.7·104 6.5·103 1.3·102 — — — — 2.2·103 5.0·103 2.7·102 — 1.0·102 3. Gams Acremonium humicola (Onions & Barron) W.2·103 7. Gams Acremonium sp.9·103 — — 9.0·102 6.1·105 1.0·103 — 2.0·103 — 3.2·103 3.0·103 — 9.0·102 3.0·103 — 3.6·103 — 9. Gams Acremonium sclerotigenum (F.7·103 4.0·103 9.3·102 1.7·103 1.1·103 2.0·104 4.2·105 5. Ames Chrysosporium indicum (Randhawa & Sandhu) Garg Chrysosporium merdarium (Link : Fries) Carmichael Chrysosporium queenslandicum Apinis & Rees Chrysosporium tropicum Carmichael Cladosporium chlorocephalum (Fresenius) Mason & M. Moreau ex Valenta) W.6·104 1.1·103 6.W. 2 Acremonium strictum W.0·103 9.0·102 7.8·103 2.B.5·103 2. Gams Acremonium chrysogenum (Thrirumalachar & Sukapure) W.9·103 — 4. pullulans Beauveria bassiana (Balsamo) Vuillemin Beauveria brongniartii (Saccardo) Petch Botryotinia fuckeliana (de Bary) Whetzel (Botrytis state) Chaetomium bostrycodes Zopf Chaetomium funicola Cooke Chaetomium globosum Kunze : Fries Chaetomium nigricolor L.8·102 — — — — 9.2·104 5. columnaris Aspergillus fumigatus Fresenius var.0·102 — 9. Saksena) Samson Alternaria alternata (Fries: Fries) von Keissler Aphanoascus terreus (Randhawa & Sandhu) Apinis (Chrysosporium state) Apiospora montanei Saccardo (Arthrinium state) Arthroderma tuberculatum Kuehn (Myceliophthora state) Ascodesmis microscopica (Crouan) Seaver Aspergillus candidus Link : Fries Aspergillus flavus Link : Fries var.7·103 4.0·102 1. Gams Acremonium persicinum (Nicot) W.5·104 — .0·102 1.9·103 1.7·103 9.0·103 — 6. ellipticus Aspergillus niger van Tiegham Aspergillus ochraceus Wilhelm Aspergillus oryzae (Ahlburg) Cohn var.5·102 1.0·103 3.3·102 3. Trotter Acremonium charticola (Lindau) W.4·103 1. fumigatus Aspergillus fumigatus Raper & Fennel var. Emmons) von Arx (Myceliophthora state) Cunninghamella elegans Lendner Cylindrocarpon sp. terreus Aspergillus versicolor (Vuillemin) Tiraboschi Aspergillus wentii Wehmer Aureobasidium pullulans (de Bary) Arnaud var.0·102 2. africanus Aspergillus terreus Thom var.B. Gams Acrodontium griseum (Fassatiova) de Hoog ` Acrophialophora fusispora (S. oryzae Aspergillus puniceus Kwon & Fennell Aspergillus sulphureus (Fresenius) Thom & Church Aspergillus terreus Fennel and Raper var.W.4·103 — 6.0·102 1.7·103 — 1.0·102 VC CFU/g dwt — — 1.0·104 9.0·102 8.2·103 — — — 6. flavus Aspergillus flavus Raper & Fennell var. Gams Acremonium fusidioides (Nicot) W. Ellis Cladosporium cladosporioides (Fresenisus) de Vries Cladosporium herbarum (Persoon : Fries) Link Cladosporium oxysporum Berkeley & Curtis Cladosporium sphaerospermum Penzig Cordyceps memorabilis Cesati (Paecilomyces state) Corynascus sepedonium (C.0·102 4.8·103 9.3·102 4. 4.36 MYCOLOGIA TABLE II.0·102 — — 4. Fungal entities isolated from compost (C) and vermicompost (VC) and their load (CFU/g dwt) expressed as the average of the highest values recorded in each of the 10 samples C CFU/g dwt 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 Absidia corymbifera (Cohn) Saccardo & A.5·102 — 3. & V.0·102 5.3·103 — 3.

Smith Emericella nidulans (Eidam) Vuillemin var. Gams & Veen baas-Rijks Mortierella humilis Linnemann ex W.0·103 b 1.1·103 4. 3 Mucor circinelloides (Hagem) Schipper f.3·104 1.7·103 7.0·103 — 1. Gams Mortierella hyalina (Harz) W.3·103 8. Graphium putredinis (Corda) S. 2 Fusarium sp.4·103 — 3.8·103 1.0·104 4.0·102 4.2·103 — 2.103 a 9.0·103 1. Huges Haematonectria haematococca (Berkeley & Broome) Samuels & Niremberg (Fusarium state) Humicola fuscoatra Traaen var. Smith Doratomyces purpureofuscus (Schweinitz : Fries) Morton & G. Gams Mortierella indohii C. thermoidea Hypocrea rufa (Person : Fries) Fries (Trichoderma state) Leptographium sp.0·102 b 3.7·102 3.0·103 7.6·103 1. fuscoatra Humicola grisea Cooney & Emerson var. griseo-cyanus Nectria sp.0·103 1.8·102 — 6.0·102 9. nidulans Engyodontium album (Limber) de Hoog Epicoccum nigrum Link Eremascus fertilis Stoppel Eurotium amstelodami Mangin Eurotium chevalieri Mangin Eurotium intermedium Blaser Eurotium montevidense (Talice & Mackinnon) Malloch & Cain Eurotium rubrum Konig et al Eutypella scoparia (Schweinitz : Fries) Ellis & Everhart (Libertella state) Exophiala moniliae de Hoog Exophiala pisciphila McGinnis & Padhye Exophiala sp.0·103 9.1·103 — 8.0·102 — 1.9·103 — 4.1·104 6.4·104 — — 6.9·103 1. Continued ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 37 C CFU/g dwt 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 Doratomyces microsporus (Saccardo) Morton & G.4·104 a — — 2.9·1033 — — 6. Neosartorya fischeri (Wehemer) Malloch & Cain var.9·104 — 6.6·104 6.1·103 — — 1. 4 Geomyces pannorum (Link) Sigler & Carmichael var.0·102 8.Y.0·102 1.7·104 7.0·103 1.0·102 — 2. Fennelia nivea (Wiley & Simmons) Samson (Aspergillus state) Fusarium oxysporum Schlechtendahl : Fries Fusarium sp. 3 Fusarium sp.3·102 2.8·103 1.0·103 7.0·103 1. 1 Mortierella sp.7·103 3.5·103 .5·102 3.0·102 7.8·103 4.5·102 — — 6. nigra Mortierella alliacea Linnemann Mortierrella alpina Peyronel Mortierella chlamydospora (Chesters) van der Plaats-Niterink Mortierella echinosphaera van der Plaats-Niterink Mortierella globalpina W.0·103 2.ANASTASI TABLE II.0·102 — 3. 2 Mortierella sp.9·104 — 2. Chien Mortierella sp.0·103 1.1·103 — — — — 2.0·103 1. Abbott (Scopulariopsis state) Microascus cirrosus Curzi Microascus manginii (Loubiere) Curzi (Scopulariopsis state) ` Moniliella suaveolens (Burri & Staub) de Hoog var.7·103 6. Leptosphaeria coniothyrium (Fuckel) Saccardo (Coniothyrium state) Malbranchea cinnamomea (Libert) van Oorschot & de Hoog Microascus brevicaulis S. 1 Fusarium sp.0·102 — 6.0·102 — — 9. fischeri Neosartorya spinosa (Raper & Fennell) Kozakiewicz Paecilomyces variotii Bainier 1.8·102 7. pannorum Geotrichum sp.0·102 — 10.4·103 2.9·102 4.8·102 — — 1.7·102 5.1·103 2.0·103 — — — 9. Gilmaniella macrospora Moustafa Gliocladium sp.7·103 4.0·103 9.P.7.8·103 VC CFU/g dwt — 9.3·102 — — — 4.

0·103 2.4·103 1.9·102 — — 8.8·103 2. Plectosporium tabacinum (van Beyma) M.2·103 4.3·103 7. italicum Penicillium janczewskii Zaleski Penicillium jensenii Zaleski Penicillium minioluteum Dierckx Penicillium ochrochloron Biourge Penicillium paxilli Bainer Penicillium piceum Raper & Fennel Penicillium purpurescens (Sopp) Raper & Thom Penicillium purpurogenum Stoll Penicillium restrictum Gilmann & Abbott Penicillium rolfsii Thom var. verrucosum Penicillium waksmanii Zaleski Phialemonium obovatum W.7·103 9. exigua ´ Phoma sp.0·102 2. W.1·103 — 4.9·103 3. Pseudallescheria boydii (Shear) McGinnis et al Pseudallescheria boydii (Shear) McGinnis et al (Scedosporium state) Pseudogymnoascus roseus Raillo (Geomyces state) Rhizopus oryzae Went & Prinsen Geerligs Rollandina capitata Patouillard Scopulariopsis brumptii Salvanet-Duval Scopulariopsis koningii (Oudemans) Vuillemin Scopulariopsis sphaerospora Zach 1.4·103 4.0·104 1.1·102 — — 4.9·103 1.8·102 1.0·103 . Phoma exigua Desmazieres var.8·102 1.0·104 2.0·103 8.0·102 3.2·103 b 9.0·103 6.3·103 9.3·102 — — — — 7.1·104 3.0·103 1.0·102 3.0·103 9.0·102 2.3·102 4. echinulatum ` Penicillium expansum Link Penicillium glabrum (Wehmer) Westling Penicillium glandicola (Oudemans) Seifert & Samson Penicillium herquei Bainier & Sartory Penicillium implicatum Biourge Penicillium islandicum Sopp Penicillium italicum Wehmer var.5·103 6.9·104 a 2.8·102 8.0·103 1.0·102 b 1.0·102 9.0·102 3.E.0·102 1.5·102 1.3·102 5.3·104 1. Gams & McGinnis Phialophora cyclaminis van Beyma Phialophora hoffmannii group (van Beyma) Schol-Schwarz Phialophora sp.0·104 — 2.0·103 3.7·103 — 1.3·103 1.9·102 1. Continued MYCOLOGIA C CFU/g dwt 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 Penicillium aurantiogriseum Dierckx var.0·102 — 1.5·105 — 2.5·104 9.4·103 1. rolfsii Penicillium roquefortii Thom Penicillium roseopurpureum Dierckx Penicillium rugulosum Thom Penicillium simplicissimum (Oudemans) Thom Penicillium sp. Gams & Niremberg (Fusarium state) Preussia fleischhakii (Auerswald) Cain Preussia sp.0·102 1.0·103 — 4.7·102 — — 2.4·103 4.0·103 9. Palm.1·103 1.0·104 — VC CFU/g dwt 3. 1 Penicillium spinulosum Thom Penicillium verrucosum Dierckx var.5·103 — 2.7·103 — — 1.5·103 — — 4.8·104 a — 7.0·103 — — — 7.1·102 — — 7. aurantiogriseum Penicillium brevicompactum Dierckx Penicillium canescens Sopp Penicillium chermesinum Biourge Penicillium chrysogenum Thom Penicillium citrinum Thom Penicillium dierckxii Biourge Penicillium digitatum Saccardo Penicillium diversum Raper & Fennell Penicillium echinulatum Raper & Thom ex Fassatiova var.2·105 b — — — 3.2·103 3.3·105 a 7. Phomopsis sp.0·102 1.3·102 6.1·102 1.2·104 — 2.1·103 7.7·104 3.38 TABLE II.7·103 1.4·103 2.

0·102 1.0·102 — 1.9·103 9.8·103 7. 2). Hughes Staphylotrichum coccosporum J.9·103 — — — 4.0·102 9.6·103 9.4·103 — 1.3·102 — — — 1.8·103 2.0·102 4.6·104 1. griseocyanus in VC2 (TABLE II. 0.7 104 CFU/g dwt in VC versus 2. flavus Talaromyces helicus (Raper & Fennell) C.0·102 7.7·103 9.9·103 9. This load is comparable with that observed in the richest soils (Thorn 1997) and justifies the use of C as a bioactivator in landfills.0·103 — 6.6·103 7.5·104 — — 6.0·102 7.4·103 5.5·104 4.2 105 CFU/g dwt.ANASTASI TABLE II.1·103 2. FIG.8·104 7. 2).0·104 — — 4.0·103 8.4·104 — VC CFU/g dwt — 1. Continued ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 39 C CFU/g dwt 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 197 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 Scytalidium lignicola Pesante Stachybotrys chartarum (Ehrenberg) S.8·104 9. whose species fall mainly in zone I (FIG. Meyer & Nicot Syncephalastrum racemosum Cohn ex Schroter Talaromyces flavus (Klocker) Stolk & Samson var.0·103 8.2·104 2. 3 from VC) compared with the SM morphotypes (6 from C.0·102 Different letters indicate significant differences (P compost and vermicompost. Benjamin var.0·103 7.2·103 7.0·102 — — 9.9 104 CFU/g dwt in C) (TABLE II).7·102 — 1.A. Dark SM were more varied in morphology in VC (mainly traceable in zone III) and overall load was 3 higher (8. made mainly from plant debris.5·103 1. Few basidiomycete morphotypes were isolated (2 from C.5·103 — 1.0·102 1. Mann-Whitney test) among the load of the same species in due to the genus Mortierella (10 species versus 5) (TABLE II).1·102 — 1.05.0·102 4.8·102 3. helicus Talaromyces helicus Stolk & Samson var.0·103 1. major Thermomyces lanuginosus Tsiklinsky Thielavia basicola Zopf Thielavia heterothallica von Klopotek (Myceliophthora state) Thysanophora penicilloides (Roumeguere) Kendrick ` Torrubiella confragosa Mains (Verticillium state) Trichoderma hamatum (Bonorden) Bainier Trichoderma harzianum Rifai Trichosporiella sporotrichoides van Oorschot Ulocladium alternariae (Cooke) Simmons Verticillium nigrescens Pethybridge Westerdykella dispersa (Clum) Cejp & Milko Basidiomycetes with clamp connections Basidiomycetes DBB Basidiomycetes with arthroconidia DBB Avellanea sterile mycelia Dark sterile mycelia Dark sterile mycelia with chlamydospores Dark sterile mycelia with chlamydospores in chain Dark sterile mycelia with chlamydospores in chain and setole Dark sterile mycelia with red exudate Dark sterile mycelia with sclerotia Dark sterile mycelia with setole Dark sterile mycelia with vesicles Hyaline sterile mycelia Hyaline sterile mycelia with vesicles Hyaline sterile mycelia with chlamydospores Yellow sterile mycelia Yellow sterile mycelia with red reverse 7.0 105 CFU/g dwt).9·103 6.0·102 7. though still very high . C.R.8·102 — — — 9. In VC the load was almost halved (up to 4. 14 from VC) (TABLE IV).7·103 1. Rhizopus oryzae and Absidia corymbifera were present only in C5 and C3 respectively. DISCUSSION These results contribute to the microbiological understanding of commercial composts.3·104 4.0·102 1. displayed a fungal load up to 8. whereas Cunninghamella elegans was present only in VC8–10 and Mucor circinelloides f. whose fungal component is often overlooked despite the favorable and unfavorable effects of fungi in the situations in which composts are employed.

The lower fungal density observed in VC is accompanied by a wider biodiversity. species that can survive during the preparation and life of the finished product. Several thermotolerant or thermophilic species (Domsch et al 1980) were isolated from both composts.84 1. However.29 a a a a 23. Cailleux 1973). showing both a greater species richness (Margalef index) and a greater evenness (Berger-Parker. the latter also shown by the rank abundance plot. Their overall load was about 9 105 CFU/g dwt in C and about ⅓ in VC. Fermor et al 1979. b. Diversity indices (Margalef. molecular methods identify most bacteria. Smit et al 1999). we found a substantial presence in VC of Chrysosporium and Scopulariopsis species. Moreover. as demonstrated by Roberts and collaborators (2002) in a study of an in-vessel compost and by Peters and collaborators (2000) in a study of composting of agricultural substrates. Most C and VC samples are distinguished in function of the presence of species regarded as typical of each matrix because they are present. enabling them to invade and parasitize cornified tissues (Rip- . Filipello Marchisio 2000).79 1. Tiunov and Scheu 2000). This was due to the use of three kinds of media and three incubation temperatures to increase the chances of isolating rare or less competitive species. might accumulate because thermophilic preconditioning could encourage the development and proliferation of thermotolerant or thermophilic species. time-consuming techniques that allow only investigation of the cultivable portion of the mycoflora and cannot provide a precise quantitative estimate. incompleteness of gene databases and low taxonomic resolution of DNA sequences (Anderson et al 2003. 1994a. All diversity indexes. were significantly higher in VC. Bridge et al 2003.08 8. Peters et al 2000. This substantial load. Simpson indeces) of fungal communities in compost (C) and vermicompost (VC) C Margalef DMg Berger-Parker 1/d Shannon H Simpson 1/D 18.84 b b b b Different letters indicate significant differences (P 0.05. The higher biodiversity may be due to a favorable action of earthworms (Brown 1995.01 3. 1. Cladosporium. however. molecular techniques only complement the conventional techniques that remain indispensable for the complete study of fungus communities and provide pure cultures that can be used for further physiological characterization of each isolate. VanderGheynst et al 2002.73 2. The differences in the qualitative and quantitative composition of the mycoflora in C and VC are well represented in the DCA plot. The main obstacles stem from inefficient DNA extraction. This finding is of particular interest because both species are potential human and animal pathogens.40 MYCOLOGIA TABLE III. Rank abundance plot of compost (cross) and vermicompost (triangle) fungal communities.18 2. which frequently demonstrate keratinolytic activity (Filipello Marchisio et al 1986. we found Scedosporium state of Pseudallescheria boydii and Aspergillus fumigatus. Penicillium. Abundance is the fungal load expressed as CFU/g dwt. prevalent during vermicomposting that are conducive to more types of fungi. Pseudallescheria and Thermomyces genera.56 VC 0. 1991.97 0. non-optimal primer selection. Shannon. Rapid molecular PCR-based techniques now are used to overcome the problems with cultivationbased.28 0. Shannon. Employment of a conventional isolation technique results in the identification in both composts of a huge number of species compared with similar studies (Straatsma et al 1994. also shows that some samples of both composts cannot be separated because they are composed of a similar mycoflora. Malbranchea. and greater than in many agricultural soils (Luppi Mosca et al 1976). Most of the 66 species common to both composts belong to the Acremonium. many regarded as the most common in composting materials.39 0. Simpson indexes). or to a more varied composition of the raw materials and to the mesophilic conditions FIG. Aspergillus.24 4. Berger-Parker. In our opinion. Among the more abundant species in both composts. in fact. The DCA. due to their thermotolerance and/or capacity to degrade a wide range of organic waste (Miller 1996).87 4.77 0.86 0. either exclusively or preponderantly. Mann-Whitney test) between the values of diversity indices in C and VC. produced by a mesophilic process in a compost. but only identify a few fungus species in samples from complex environments such as composts.

species exclusive of compost. TABLE IV. Scatterplot of the DCA of 10 samples of compost (circle) and 10 samples of vermicompost (square) along with 194 fungal entities (for species name refer to TABLE II). species exclusive of vermicompost. of genera 3 12 — 44 — VC No. The first two axes are shown (eigenvalues: axis 1 0. Number of fungal genera and species among different taxonomic groups in compost (C) and vermicompost (VC) and their relative load (%) C No.297). axis 2 0.576.ANASTASI ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 41 FIG. of genera Zygomycetes Ascomycetes Basidiomycetes Mitosporic fungi (including Ascomycetes in their anamorphic state) Sterile mycelia 3 7 — 37 — No. 2. of species or morphotypes 7 20 3 98 14 Load % 6 1 1 86 6 Load % 6 4 2 76 12 . of species or morphotypes 12 9 2 89 6 No. species common to both composts.

Panchal G. and for preparation of quality certificates and correct management practices to safeguard the health of compost workers and users. We are grateful to Prof. Scannerini S. Along with the systematic characterization of fungal communities in compost. as already observed by Brown (1995) and Tiunov and Scheu (2000). BioCycle Magazine. Moreover. Miserere for their assistance in the preparation of the statistics. Beffa T. Another point is the isolation of a low number of potentially phytopathogenic species from both composts. Valutazione della fitotossicita di un am` mendante compostato verde ed un ammendante compostato misto [Doctoral dissertation]. Beffa et al 1998. Med Mycol 36: 137–145. can be determined only by comparing identically composed raw materials. This too. maintain that their ingestion is impeded by protective chemical barriers. Pre- This study was financed by CEBIOVEM (Centro di Eccellenza per la Biosensoristica Tramite l’utilizzo di Organismi Vegetali e Microbici) and by Marcopolo Environmental Group which also supplied the mature composts. several fungal strains from these composts now are being investigated to test their capability to decolorize several synthetic dyes and degrade some polycyclic aromatic hydrocarbons: naphthalene. Prosser JI. however. Odds 1991). Mycological control and surveil´ lance of biological waste and compost. the presence of animal skin. Biodiversity PRO. Campbell CD. 2003. L. On the unreliability of published DNA sequences. Roberts PJ. The extent earthworms influence the development of health-threatening fungi. 1997. Italy: 123 p.42 MYCOLOGIA liminary results show that taxonomic fungal diversity reflects a different metabolic potential (Anastasi et al 2004). Caccavo D. 1999. LITERATURE CITED Airaudi D. 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