Succession of Actinomycetes During Composting Proccess of Dairy-Farm Waste Investigated by Culture-Dependent and Independent Approaches

• Main Authors: Mukhlissul Faatih1, Jaka Widada, Ngadiman N
• Format: Article
• Language: English
• Published: Universitas Gadjah Mada, Yogyakarta 2015-11-01
• Series: Indonesian Journal of Biotechnology
• Online Access: http://journal.ugm.ac.id/ijbiotech/article/view/7799

Mesophilic, thermophilic, and maturation phases were recognized in composting proccess. Temperature<br />changes influence the microbial communities in compost within composting proccess. Actinomycetes account for a<br />larger part of compost microbial population. The aim of this research was to study succession of actinomycetes<br />community during composting of dairy-farm waste investigated by culture-dependent and independent<br />approaches.<br />In culture-independent method, the succession of actinomycetes community was analyzed by nestedpolymerase<br />chain reaction of ribosomal intergenic spacer (nested-PCR RISA) using spesific primer F243 and primer<br />R23S followed by a second PCR using primers F968 and R23S. In culture-dependent method actinomycetes from<br />compost were isolated on selective media, starch-nitrate medium and humic-acid + vitamins medium. DNA of<br />actinomycetes was extracted and amplified by repetitive sequence-based PCR (rep-PCR) using primer BOXA1R. The<br />banding patterns were used to generate dendrograms by UPGMA clustering with NTSYS program. Microcosm<br />containing sterile rice-straw and water which is inoculated with each actinomycetes isolates was used for examining<br />the ability of each isolate in rice-straw degradation.<br />The experiment results showed that succession of both bacteria and actinomycetes was occured within<br />composting proccess of dairy-farm waste. Analysed by culture-independent method revealed that the highest<br />community of compost&rsquo;s bacteria was on mesophilic, thermophilic, and maturation phases, respectively. Whereas<br />PCR-nested RISA resulted the highest population of actinomycetes was on thermophilic, maturation, and mesophilic<br />phases, respectively. By culture-dependent method was obtained 29 actinomycetes isolates from mesophilic phase,<br />23 isolates from thermophilic phase, and 19 isolates from maturation phase. Genetic diversity analysis of the obtained<br />isolates showed the presence of phylogenetic grouping on each phase of composting proccess. This result illustrated<br />the occurance of succession of actinomycetes community in compost. The ability of each isolates in rice-straw<br />degradation was different, and SnT9 isolate was found to be a promising rice-straw degrader.<br />Keywords: succession, actinomycetes, composting, nested-PCR RISA, rep-PCR