title: Characterization of cultivated fungi isolated from grape marc wastes through the use of amplified rDNA restriction analysis and sequencing.
abstract: Microbial assessment of grape marc wastes, the residual solid by-product of the wine-industry, was performed by identifying phylogenetically the fungal culturable diversity in order to evaluate environmental and disposal safety issues and to discuss ecological considerations of applications on agricultural land. Fungal spores in grape marc were estimated to 4.7 x 10(6) per g dry weight. Fifty six fungal isolates were classified into eight operational taxonomic units (OTUs) following amplified ribosomal DNA restriction analysis (ARDRA) and colony morphology. Based on 18S rRNA gene and 5.8S rRNA gene-ITS sequencing, the isolates representing OTUs #1, #2, #3, and #4, which comprised 44.6%, 26.8%, 12.5%, and 5.3%, respectively, of the number of the total isolates, were identified as Aspergillus fumigatus, Bionectria ochroleuca, Haematonectria haematococca, and Trichosporon mycotoxinivorans. The isolates of OTU#5 demonstrated high phylogenetic affinity with Penicillium spp., while members of OTUs #6 and #7 were closer linked with Geotrichum candidum var. citri-aurantii and Mycocladus corymbifer, respectively (95.4 and 97.9% similarities in respect to their 5.8S rRNA gene-ITS sequences). The OTU#8 with a single isolate was related with Aspergillus strains. It appears that most of the fungal isolates are associated with the initial raw material. Despite the fact that some of the species identified may potentially act as pathogens, measures such as the avoidance of maintaining large and unprocessed quantities of grape marc wastes in premises without adequate aeration, together with its suitable biological treatment (e.g., composting) prior to any agriculture-related application, could eliminate any pertinent health risks.
authors.affiliation: National Agricultural Research Foundation, Institute of Kalamata, Lakonikis 87, 24100, Kalamata, Greece.
authors.forename: Spyridon
authors.initials: S
authors.lastname: Ntougias
authors.forename: Nektarios
authors.initials: N
authors.lastname: Kavroulakis
authors.forename: Kalliope K
authors.initials: KK
authors.lastname: Papadopoulou
authors.forename: Constantinos
authors.initials: C
authors.lastname: Ehaliotis
authors.forename: Georgios I
authors.initials: GI
authors.lastname: Zervakis
mesh_terms: D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D017753:Ecosystem; D005658:Fungi / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D005800:Genes, Fungal; D007220:Industrial Waste; D010802:Phylogeny; D012037:Refuse Disposal; D012988:Soil Microbiology; D027843:Vitis / Q000382:microbiology*; D014920:Wine
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D007220:Industrial Waste