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

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. Compost (C) was produced in an outdoor pile (3 m wide. SM with RESULTS The total fungal load was high in both composts: from 5.9%). As diversity increases. d and D decrease. the moisture content 38. they were transferred to the media recommended by the authors of selected genus monographs for species identification. A 10 g portion of each sample was suspended in 90 ml Na4P2O7·10 H2O to disperse organic colloids. and 4 to 4. A total of 194 fungal entities were identified from the two composts. depending on media or incubation temperature (TABLE I). 1. poultry and various zoo animals) and 30% plant debris from various sources. von Arx 1981. humic organic carbon (humic acid fulvic acid) 7. allow isolation of slow-growing colonies and focus on fungi of medical interest (Airaudi and Filipello Marchisio 1996. Employment of the CMC and CX media. The final dilution (1: 20 000) was plated (1 ml per plate) on 11 replicates: five of potato-dextrose agar (PDA). The Mann-Whitney test (StatView 1988) was run to assess the significance (P 0. The pH of the final product was 7. VC1–10). Shannon. The culture and/or incubation conditions produced different load values within the same compost. higher temperatures.05) of the differences between the two composts (total load. of which 118 came from C and 142 from VC.9. Vermicompost (VC) was produced in an outdoor pile (3 m wide. The greatest number of species were isolated from both composts on PDA incubated at 24 C. S is the number of fungal entities and N is the total number of individuals).2. Only 66 were common to both composts (TABLE II). were examined according to the guidelines proposed by the Piedmont Region (Trombetta et al 1998). cycloheximide and carboxy-methyl cellulose all reduced CFU/g dwt values.34 MYCOLOGIA clamp connections or positive to the reaction with Diazonium Blue B salts (DBB). species and genera load) and between all treatments (three media and three incubation temperatures) in the composts. We therefore used these indexes in their reciprocal form 1/d and 1/D (Magurran 1988). a significant reduction in CFU/g dwt was induced by higher incubation temperatures and the addition of cycloheximide. needed to determine its quality and field of application (Peters et al 2000). according to Summerbell (1985). Fungi were identified conventionally according to their macroscopic and microscopic features. three of carboxy-methyl cellulose agar (CMC) and three of PDA supplemented with cycloheximide (CX) to retard the growth of all fungi.0 from 5. Simpson indexes) were applied to assess biodiversity (Biodiversity PRO 1997). 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. Kiffer and Morelet 1997). Sterile mycelia (SM) were classified according to their hyphal pigments and their production of chlamydospores. and incubation at 37 C and 45 C allowed . during which the piles were turned periodically by machine. In C. unless denoted otherwise. humic organic carbon (humic acid fulvic acid) 3. were classified as basidiomycetes. temperatures and moisture measurements were taken outdoors and are approximations. earthworms (Lumbricus rubellus Hoffmeister) were added (50 103 worms per m3 organic matter) and the pile was turned periodically by machine. 50 m long.6% of the dry matter and the C/N ratio 15.2 105 CFU/g dwt in C. sclerotia or vesicles. After preconditioning for several days. The nonparametric Mann-Whitney test for independent groups (StatView 1988) was run to assess the significance (P 0. Plates were incubated at 24 C.05) of the differences of each index between the two composts. 0. the Berger-Parker index from the formula d Nmax/N (where Nmax is the number of individuals in the most abundant species).5%.5 m high) composed of 70% dung (from cows. During this mesophilic phase the temperature never exceeded 25–30 C. All statistics were obtained from the highest load of each species in the 9 treatments of each sample. the two population structures were analyzed with the rank-abundance plot (Biodiversity PRO 1997). Diversity indexes based on species richness (Margalef index) and species relative abundance (BergerParker.0 104 to 8. Hanlin 1990. the Margalef index was calculated from the formula DMg (S 1)/ln N (here and throughout. In VC. MATERIALS AND METHODS The following physical dimensions. the Simpson index from the formula D {[ni(ni 1)]/[N(N 1)]}.3 10 105 CFU/g dwt in VC. After determination of their genera (Domsch et al 1980. The maximum temperature during the composting was 60 C. except for CMC at 45 C and CX at all temperatures (TABLE I). further dilutions were made in NaCl (0. the Shannon index from the formula H Spi(lnpi) (where pi is the proportion of individuals found in the ith species). Both composts were stored in polypropylene bags 1–3 mo at 10 C before being sold. however. Ten approximately 1 kg samples per compost (C1–10. Moreover.6% of the dry matter and the C/N ratio about 25.5 m high) from plant debris from various sources by a conventional thermophilic process that lasted ca 6 mo. the moisture content 40%. 37 C and 45 C to isolate mesophilic and thermotolerant/ thermophilic fungi with the result that 33 replicates were made for each sample. 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 load values in C always were higher. Filipello Marchisio et al 1996). The pH of the final product was 7. According to Magurran (1988). during which the temperature rose to 60 C. 50 m long.

The 194 fungal entities comprised 117 mitosporic fungi. and Trichoderma species (8. mainly . zone III containing most of the VC samples (VC1.5·105 9. of fungal entities 70 42 43 32 17 9 25 5 3 (31) (6) (7) (5) (3) (0) (3) (0) (0) 4.3 105 CFU/g dwt) and was associated with it in the DCA and included in zone I (FIG. 45 ascomycetes.0·105 2.1·104 2. 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. 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.ANASTASI ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 35 TABLE I.1 104 in VC versus 1. aurantiogriseum (P 0. aurantiogriseum var. CMC.6·105 1. namely the Scedosporium state of Pseudallescheria boydii and Aspergillus fumigatus. The total load of Penicillium was 3. and Talaromyces flavus var. There were no significant differences between composts in the quantitative composition of the two sets of Cladosporium and Acremonium species (about 5.0 104 and 1.4·105 5. fumigatus (TABLE II).2 105 CFU/g dwt in C. and many species solely were present in C or in VC (TABLE II).2 104 in VC versus 0 CFU/g dwt in C. 2. 8–10) and the entities equally distributed between C and VC.2 103 CFU/g dwt) were present exclusively in C. The load of zygomycetes was similar in C and VC with greater species diversity in C (TABLE IV).8 105 CFU/g dwt in C and 1. 15 zygomycetes. The Aspergillus load was not significantly different: 1.1 103 CFU/g dwt in VC). the isolation of a good number of species that otherwise would have been missed: 24 from C and 30 from VC (TABLE I). zone II containing C and VC samples (C1–3 e VC3. They included Aspergillus fumigatus var. 4–7) and the entities found only or preponderant in VC.03) showed prevalence in C and associated with C8 and C5 respectively (FIG. and 3.1·105 1.05.0 105 CFU/g dwt in VC and 1. 2).3·105 7.5·105 9. though P.0·104 4. whereas Fusarium species prevailed in C (3. 2).5·104 9. CX) incubated at 24 C.8·105 6. Absidia corymbifera alone was isolated from C only.013) and P. The number of species and the load of ascomycetes were higher in VC (TABLE IV).0·104 1.1·105 3.4·105 4. 2). 1). The DCA scatterplot (FIG. which demonstrates the quantitative domination of two species.0·104 a* a* b* c* ac* bc c c bc No. Mean fungal load (CFU/g dwt SE) and number of fungal entities isolated in compost (C) and vermicompost (VC) on 3 media (PDA.3·104 VC 8. Chrysosporium and Scopulariopsis species prevailed in VC (respectively 1.0·104 2. the greater number of species corresponds to higher biodiversity index values (TABLE III).105 CFU/g dwt in VC. Both loads were composed mainly of thermotolerant A. flavus (mainly present in VC2. The genera with the highest load and number of species in both composts were Penicillium and Aspergillus.9·104 1. 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. which displayed high loads in all the samples and was located in zone II (FIG.7·104 2.2·105 8. 37 C. the thermotolerant fungus Scedosporium state of Pseudallescheria boydii displayed a significantly greater load (P 0.2·105 1.8·105 2. This difference is not signif- icant. or found only in C or in VC.1 104 in C versus 2.5·105 1. 2) shows the distribution of the samples and the 194 fungal entities.0 104 CFU/g dwt respectively in both composts).9·104 5. 2). In VC.9·104 4. particularly owing to the presence of Corynascus sepedonium (mainly present in VC3). 45 C. 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.4·105 1.10) (TABLE II. Both composts were dominated by mitosporic fungi (including the ascomycetes in their anamorphic state) (TABLE IV).5·104 5. roseopurpureum (P 0. Malbranchea cinnamomea. 2). 14 SM morphotypes and three basidiomycete morphotypes (TABLE II).3. Of the most abundant species in both composts. mainly in C4 (FIG. ellipticus.6·104 2.1·104 a* a* b* a* ab* bc d d c No. Paecilomyces variotii and Thermomyces lanuginosus.1·104 4. 2).6·105 2.0·104 3.0012) in C (7. but present in smaller quantities and thus regarded as occasionals (FIG.0·105 2. 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. The lower evenness of C is illustrated in the rank abundance plot (FIG.7 104 CFU/ g dwt in C).3·105 1. fumigatus var. Eurotium chevalieri (mainly present in VC6).8·104 6.6·105 1. FIG.

W.0·103 — 2.3·102 3.0·103 2.9·103 1.0·102 4.0·102 1.0·102 7.8·102 1. Emmons) von Arx Corynascus sepedonium (C.7·103 1.7·103 — 1.0·103 — 6.3 102 — — 7.0·102 VC CFU/g dwt — — 1.2·103 — — — 6.0·102 1.5·104 — .4·103 1.3·102 1. 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.0·103 2.7·103 4.1·104 1.2·103 3.0·103 9. Moreau ex Valenta) W. 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.0·103 — 3.B.0·102 6.3·102 — — — — 2.4·103 1.3·103 — 3.7·104 6.3·103 6.2·103 5.7·103 9.2·104 5.0·103 9.5·103 1. Gams Acremonium humicola (Onions & Barron) W.3·102 1.0·102 5.0·103 — 9. Trotter Acremonium charticola (Lindau) W.5·103 2.6·104 1. africanus Aspergillus terreus Thom var.6·103 — 9.7·103 4.0·104 9.0·102 3. oryzae Aspergillus puniceus Kwon & Fennell Aspergillus sulphureus (Fresenius) Thom & Church Aspergillus terreus Fennel and Raper var. 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.B.0·102 1. ellipticus Aspergillus niger van Tiegham Aspergillus ochraceus Wilhelm Aspergillus oryzae (Ahlburg) Cohn var.4·103 — 6.2·103 7. Gams Acremonium sp.1·105 1.1·103 2.9·103 1.0·102 3.9·103 — — 9.0·102 2.5·102 1. Gams Acremonium sclerotigenum (F.8·103 2. & V.8·103 9.0·103 9. 4. flavus Aspergillus flavus Raper & Fennell var. Gams Acremonium persicinum (Nicot) W.1·103 6. Ames Chrysosporium indicum (Randhawa & Sandhu) Garg Chrysosporium merdarium (Link : Fries) Carmichael Chrysosporium queenslandicum Apinis & Rees Chrysosporium tropicum Carmichael Cladosporium chlorocephalum (Fresenius) Mason & M.0·102 3.W.0·102 — 9.9·103 — 4.0·103 3.0·103 — 3. 1 Acremonium sp. 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.7·102 — 1. Gams Acrodontium griseum (Fassatiova) de Hoog ` Acrophialophora fusispora (S.0·102 8. terreus Aspergillus versicolor (Vuillemin) Tiraboschi Aspergillus wentii Wehmer Aureobasidium pullulans (de Bary) Arnaud var. fumigatus Aspergillus fumigatus Raper & Fennel var.3·102 4.0·102 — — 4.5·102 — 3.2·105 5.36 MYCOLOGIA TABLE II.8·102 — — — — 9. columnaris Aspergillus fumigatus Fresenius var. Gams Acremonium fusidioides (Nicot) W. Emmons) von Arx (Myceliophthora state) Cunninghamella elegans Lendner Cylindrocarpon sp. Gams Acremonium chrysogenum (Thrirumalachar & Sukapure) W. 2 Acremonium strictum W.0·104 4.

4·103 — 3. Gams Mortierella hyalina (Harz) W.8·102 — — 1.ANASTASI TABLE II.6·104 6. fischeri Neosartorya spinosa (Raper & Fennell) Kozakiewicz Paecilomyces variotii Bainier 1.103 a 9.0·102 — 2. Smith Emericella nidulans (Eidam) Vuillemin var.Y.3·102 2.0·103 1.3·103 8.0·102 — 1. fuscoatra Humicola grisea Cooney & Emerson var.7·103 7.1·104 6. 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.8·102 — 6.0·102 — — 9.0·103 1. pannorum Geotrichum sp. Fennelia nivea (Wiley & Simmons) Samson (Aspergillus state) Fusarium oxysporum Schlechtendahl : Fries Fusarium sp.7·102 3.9·104 — 2.0·103 1. Smith Doratomyces purpureofuscus (Schweinitz : Fries) Morton & G.9·1033 — — 6.0·103 9. Abbott (Scopulariopsis state) Microascus cirrosus Curzi Microascus manginii (Loubiere) Curzi (Scopulariopsis state) ` Moniliella suaveolens (Burri & Staub) de Hoog var. 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.8·103 1. Graphium putredinis (Corda) S.0·103 — 1. 4 Geomyces pannorum (Link) Sigler & Carmichael var.1·103 — 8. Leptosphaeria coniothyrium (Fuckel) Saccardo (Coniothyrium state) Malbranchea cinnamomea (Libert) van Oorschot & de Hoog Microascus brevicaulis S.4·104 — — 6. 3 Fusarium sp. 1 Fusarium sp. Gilmaniella macrospora Moustafa Gliocladium sp.9·103 1.0·103 b 1.0·102 — 3.1·103 — — 1.9·102 4. 3 Mucor circinelloides (Hagem) Schipper f.8·103 1.0·103 2. Gams & Veen baas-Rijks Mortierella humilis Linnemann ex W.0·102 9.8·103 VC CFU/g dwt — 9.1·103 2.9·103 — 4. griseo-cyanus Nectria sp.7·103 3.0·103 7.5·102 3.4·103 2.7·103 4.7·104 7.0·102 4. 2 Mortierella sp. Gams Mortierella indohii C.7.1·103 — — — — 2.0·102 1.0·102 7.7·103 6. Huges Haematonectria haematococca (Berkeley & Broome) Samuels & Niremberg (Fusarium state) Humicola fuscoatra Traaen var.0·103 1.4·104 a — — 2.5·102 — — 6.2·103 — 2.8·102 7. Neosartorya fischeri (Wehemer) Malloch & Cain var.0·103 1.0·102 b 3.9·104 — 6. thermoidea Hypocrea rufa (Person : Fries) Fries (Trichoderma state) Leptographium sp.0·103 9.0·102 — 10. Chien Mortierella sp. 2 Fusarium sp.7·102 5.3·104 1.P. nigra Mortierella alliacea Linnemann Mortierrella alpina Peyronel Mortierella chlamydospora (Chesters) van der Plaats-Niterink Mortierella echinosphaera van der Plaats-Niterink Mortierella globalpina W.0·102 8. 1 Mortierella sp.5·103 .0·103 1.3·102 — — — 4.0·102 — 6.0·103 — — — 9.0·104 4.8·103 4.0·103 7.6·103 1.1·103 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.8·102 8.2·104 — 2.0·103 .7·103 — — 1. rolfsii Penicillium roquefortii Thom Penicillium roseopurpureum Dierckx Penicillium rugulosum Thom Penicillium simplicissimum (Oudemans) Thom Penicillium sp.4·103 2.0·102 3.0·103 9.7·102 — — 2.0·104 2.5·104 9. Phoma exigua Desmazieres var.7·103 1.3·103 7.8·104 a — 7.0·103 9.7·103 9.0·102 2.9·102 1.3·103 1.38 TABLE II.0·102 b 1.8·102 1.0·104 — VC CFU/g dwt 3.4·103 4.1·104 3.0·102 3.3·102 — — — — 7.1·102 — — 4. W.7·103 — 1.E.0·103 — 4.2·103 3.5·103 6.2·105 b — — — 3.0·102 2.8·103 2.1·103 7.0·102 — 1.5·105 — 2.3·103 9.8·102 1.9·104 a 2.3·105 a 7.0·103 6.9·102 — — 8.5·103 — — 4.0·103 — — — 7.0·103 1.0·103 8.1·102 1.0·103 3. 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.7·104 3.0·102 3.0·102 1.4·103 4. Plectosporium tabacinum (van Beyma) M.0·102 9.5·103 — 2.3·102 5.0·102 1. 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. exigua ´ Phoma sp. Palm.4·103 1. Phomopsis sp.1·103 1. 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. Gams & Niremberg (Fusarium state) Preussia fleischhakii (Auerswald) Cain Preussia sp.3·104 1.0·104 — 2. 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.0·103 1.0·103 2.2·103 b 9.4·103 1.9·103 1.0·104 1.3·102 4.5·102 1. verrucosum Penicillium waksmanii Zaleski Phialemonium obovatum W.3·102 6.1·102 — — 7.0·102 1. Gams & McGinnis Phialophora cyclaminis van Beyma Phialophora hoffmannii group (van Beyma) Schol-Schwarz Phialophora sp.9·103 3. 1 Penicillium spinulosum Thom Penicillium verrucosum Dierckx var.2·103 4.1·103 — 4.

5·104 — — 6.7 104 CFU/g dwt in VC versus 2. In VC the load was almost halved (up to 4. 3 from VC) compared with the SM morphotypes (6 from C.6·103 7.9·103 6.0·102 1. whereas Cunninghamella elegans was present only in VC8–10 and Mucor circinelloides f.0·102 1.7·103 9.A.R.2·104 2. Rhizopus oryzae and Absidia corymbifera were present only in C5 and C3 respectively. This load is comparable with that observed in the richest soils (Thorn 1997) and justifies the use of C as a bioactivator in landfills. DISCUSSION These results contribute to the microbiological understanding of commercial composts.9·103 9.4·103 — 1.0·103 — 6.5·103 — 1. though still very high .1·102 — 1.8·103 2.9 104 CFU/g dwt in C) (TABLE II).0·102 — — 9.8·102 3.8·104 7.0·102 — 1.0·103 8. C.0·102 4. FIG.0·102 7.0·103 1.0·102 4.4·103 5.8·103 7. 14 from VC) (TABLE IV).0·103 8. 0.ANASTASI TABLE II.8·104 9.7·103 1. 2). Hughes Staphylotrichum coccosporum J.3·104 4.7·102 — 1.6·103 9.1·103 2. Mann-Whitney test) among the load of the same species in due to the genus Mortierella (10 species versus 5) (TABLE II).0·102 1.5·104 4.2·103 7.0·102 Different letters indicate significant differences (P compost and vermicompost. 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. displayed a fungal load up to 8.0·104 — — 4. Dark SM were more varied in morphology in VC (mainly traceable in zone III) and overall load was 3 higher (8. Few basidiomycete morphotypes were isolated (2 from C.0·103 7. whose fungal component is often overlooked despite the favorable and unfavorable effects of fungi in the situations in which composts are employed. whose species fall mainly in zone I (FIG.05.9·103 — — — 4.9·103 9.3·102 — — — 1. made mainly from plant debris.0 105 CFU/g dwt).0·102 7.8·102 — — — 9. griseocyanus in VC2 (TABLE II. Meyer & Nicot Syncephalastrum racemosum Cohn ex Schroter Talaromyces flavus (Klocker) Stolk & Samson var. Benjamin var.5·103 1.2 105 CFU/g dwt. 2). helicus Talaromyces helicus Stolk & Samson var.6·104 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. flavus Talaromyces helicus (Raper & Fennell) C.0·102 7.0·102 9.4·104 — VC CFU/g dwt — 1.

Tiunov and Scheu 2000). Most C and VC samples are distinguished in function of the presence of species regarded as typical of each matrix because they are present. due to their thermotolerance and/or capacity to degrade a wide range of organic waste (Miller 1996). molecular methods identify most bacteria. Malbranchea. Aspergillus.97 0. The DCA.05. Simpson indexes). but only identify a few fungus species in samples from complex environments such as composts.84 b b b b Different letters indicate significant differences (P 0. Rapid molecular PCR-based techniques now are used to overcome the problems with cultivationbased. This substantial load. 1994a. and greater than in many agricultural soils (Luppi Mosca et al 1976). species that can survive during the preparation and life of the finished product. Moreover. In our opinion. Their overall load was about 9 105 CFU/g dwt in C and about ⅓ in VC. Berger-Parker. Rank abundance plot of compost (cross) and vermicompost (triangle) fungal communities. 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. we found Scedosporium state of Pseudallescheria boydii and Aspergillus fumigatus. Penicillium. Pseudallescheria and Thermomyces genera. Filipello Marchisio 2000).08 8. Shannon.84 1.77 0. 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. which frequently demonstrate keratinolytic activity (Filipello Marchisio et al 1986. Simpson indeces) of fungal communities in compost (C) and vermicompost (VC) C Margalef DMg Berger-Parker 1/d Shannon H Simpson 1/D 18.86 0. incompleteness of gene databases and low taxonomic resolution of DNA sequences (Anderson et al 2003.29 a a a a 23. The lower fungal density observed in VC is accompanied by a wider biodiversity.87 4. Several thermotolerant or thermophilic species (Domsch et al 1980) were isolated from both composts.01 3. Peters et al 2000. however. 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. Fermor et al 1979.40 MYCOLOGIA TABLE III. However. enabling them to invade and parasitize cornified tissues (Rip- . Cladosporium. 1. This finding is of particular interest because both species are potential human and animal pathogens.73 2. prevalent during vermicomposting that are conducive to more types of fungi. VanderGheynst et al 2002. The differences in the qualitative and quantitative composition of the mycoflora in C and VC are well represented in the DCA plot. or to a more varied composition of the raw materials and to the mesophilic conditions FIG. Most of the 66 species common to both composts belong to the Acremonium. All diversity indexes.24 4. The higher biodiversity may be due to a favorable action of earthworms (Brown 1995. 1991. b. Diversity indices (Margalef.39 0. time-consuming techniques that allow only investigation of the cultivable portion of the mycoflora and cannot provide a precise quantitative estimate. non-optimal primer selection. Mann-Whitney test) between the values of diversity indices in C and VC. we found a substantial presence in VC of Chrysosporium and Scopulariopsis species. also shows that some samples of both composts cannot be separated because they are composed of a similar mycoflora. the latter also shown by the rank abundance plot. showing both a greater species richness (Margalef index) and a greater evenness (Berger-Parker. in fact. were significantly higher in VC. 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.18 2. might accumulate because thermophilic preconditioning could encourage the development and proliferation of thermotolerant or thermophilic species. The main obstacles stem from inefficient DNA extraction. Shannon. Smit et al 1999).79 1. Abundance is the fungal load expressed as CFU/g dwt. Among the more abundant species in both composts. many regarded as the most common in composting materials. Bridge et al 2003.28 0. either exclusively or preponderantly. Cailleux 1973). produced by a mesophilic process in a compost.56 VC 0.

2. of genera Zygomycetes Ascomycetes Basidiomycetes Mitosporic fungi (including Ascomycetes in their anamorphic state) Sterile mycelia 3 7 — 37 — No. species common to both composts. species exclusive of compost. species exclusive of vermicompost. of genera 3 12 — 44 — VC No. 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). 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. TABLE IV. of species or morphotypes 12 9 2 89 6 No. of species or morphotypes 7 20 3 98 14 Load % 6 1 1 86 6 Load % 6 4 2 76 12 .297).576. axis 2 0.ANASTASI ET AL: FUNGAL COMMUNITIES IN COMPOST AND VERMICOMPOST 41 FIG.

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