Bacterial Groups Concerned with Maturing Process in Manure Production Analyzed by a Method Based on Restriction Fragment Length Polymorphism Analysis

Katsuji Watanabe^1*, Naoto Horinishi¹, Kunimasa Matumoto¹, Akihiro Tanaka², Kenichi Yakushido³

¹Department of Life, Environment and Materials Science, Fukuoka Institute of Technology, Fukuoka, Japan.
²National Agricultural Research Center for Kyushu-Okinawa Region, National Agriculture and Food Research Organization, Koshi, Japan.
³National Agricultural Research Center, National Agriculture and Food Research Organization, Tsukuba, Japan.

DOI: 10.4236/aim.2015.513088                

Abstract

Composting is a biological aerobic decomposition process consisted from different phases. Although the Japanese Standards for manure recommended that it took at least 6 months to complete the maturing phase, there was no reliable ground. In order to find out shortening method of the maturing phase, the microorganisms concerned with a progress of the maturing was determined by using the most probable number method (MPN) and PCR-RFLP of the 16S rDNA, which was found effective to provide numbers and taxonomy of polymyxin B resistant bacterial groups in the former paper [1]. Compared to the numbers after thermophilic phase, those of Actinobacteria, δ-proteobacteria, and the other gram negative bacteria increased to 50 times, 20 times, and 10⁵ times, respectively, after maturing phase, while those of Bacillus spp., and α and β-proteobacteria decreased to 1/10, and 1/10⁵ after maturing phase. Numbers of the other Fumicutes, and γ-proteobacteria remained in the same revel. Actinobacteria, δ-proteobacteria, and the other gram negative bacteria might be concerned with a progress of the maturing phase, because these bacterial groups were detected and enumerated due to their proliferation ability. Although number of Acitinobacteria might be underestimated because of a PCR bias, the method was found effective for the purpose to monitor bacteria actively proliferated in culture medium.

Keywords

Maturing Phase, Manure Production, Microchip Electrophoresis, Multiple Enzyme Restriction Fragment Length Polymorphism Analysis, The Most Probable Number Method

Conflicts of Interest

The authors declare no conflicts of interest.

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