Succession of Actinomycetes During Composting Proccess of Dairy-Farm Waste Investigated by Culture-Dependent and Independent Approaches
https://doi.org/10.22146/ijbiotech.7799 Mukhlissul Faatih(1*), Jaka Widada(2), N. Ngadiman(3)
(1)
(2)
(3)
(*) Corresponding Author
Abstract
Mesophilic, thermophilic, and maturation phases were recognized in composting proccess. Temperature
changes influence the microbial communities in compost within composting proccess. Actinomycetes account for a
larger part of compost microbial population. The aim of this research was to study succession of actinomycetes
community during composting of dairy-farm waste investigated by culture-dependent and independent
approaches.
In culture-independent method, the succession of actinomycetes community was analyzed by nestedpolymerase
chain reaction of ribosomal intergenic spacer (nested-PCR RISA) using spesific primer F243 and primer
R23S followed by a second PCR using primers F968 and R23S. In culture-dependent method actinomycetes from
compost were isolated on selective media, starch-nitrate medium and humic-acid + vitamins medium. DNA of
actinomycetes was extracted and amplified by repetitive sequence-based PCR (rep-PCR) using primer BOXA1R. The
banding patterns were used to generate dendrograms by UPGMA clustering with NTSYS program. Microcosm
containing sterile rice-straw and water which is inoculated with each actinomycetes isolates was used for examining
the ability of each isolate in rice-straw degradation.
The experiment results showed that succession of both bacteria and actinomycetes was occured within
composting proccess of dairy-farm waste. Analysed by culture-independent method revealed that the highest
community of compost’s bacteria was on mesophilic, thermophilic, and maturation phases, respectively. Whereas
PCR-nested RISA resulted the highest population of actinomycetes was on thermophilic, maturation, and mesophilic
phases, respectively. By culture-dependent method was obtained 29 actinomycetes isolates from mesophilic phase,
23 isolates from thermophilic phase, and 19 isolates from maturation phase. Genetic diversity analysis of the obtained
isolates showed the presence of phylogenetic grouping on each phase of composting proccess. This result illustrated
the occurance of succession of actinomycetes community in compost. The ability of each isolates in rice-straw
degradation was different, and SnT9 isolate was found to be a promising rice-straw degrader.
Keywords: succession, actinomycetes, composting, nested-PCR RISA, rep-PCR
changes influence the microbial communities in compost within composting proccess. Actinomycetes account for a
larger part of compost microbial population. The aim of this research was to study succession of actinomycetes
community during composting of dairy-farm waste investigated by culture-dependent and independent
approaches.
In culture-independent method, the succession of actinomycetes community was analyzed by nestedpolymerase
chain reaction of ribosomal intergenic spacer (nested-PCR RISA) using spesific primer F243 and primer
R23S followed by a second PCR using primers F968 and R23S. In culture-dependent method actinomycetes from
compost were isolated on selective media, starch-nitrate medium and humic-acid + vitamins medium. DNA of
actinomycetes was extracted and amplified by repetitive sequence-based PCR (rep-PCR) using primer BOXA1R. The
banding patterns were used to generate dendrograms by UPGMA clustering with NTSYS program. Microcosm
containing sterile rice-straw and water which is inoculated with each actinomycetes isolates was used for examining
the ability of each isolate in rice-straw degradation.
The experiment results showed that succession of both bacteria and actinomycetes was occured within
composting proccess of dairy-farm waste. Analysed by culture-independent method revealed that the highest
community of compost’s bacteria was on mesophilic, thermophilic, and maturation phases, respectively. Whereas
PCR-nested RISA resulted the highest population of actinomycetes was on thermophilic, maturation, and mesophilic
phases, respectively. By culture-dependent method was obtained 29 actinomycetes isolates from mesophilic phase,
23 isolates from thermophilic phase, and 19 isolates from maturation phase. Genetic diversity analysis of the obtained
isolates showed the presence of phylogenetic grouping on each phase of composting proccess. This result illustrated
the occurance of succession of actinomycetes community in compost. The ability of each isolates in rice-straw
degradation was different, and SnT9 isolate was found to be a promising rice-straw degrader.
Keywords: succession, actinomycetes, composting, nested-PCR RISA, rep-PCR
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PDF DOI: https://doi.org/10.22146/ijbiotech.7799
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