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3 メ タ ン発 酵 に お け る硫 酸還 元 菌 とC02/H2資 Photo.1 Colony vated Photo.2Colony of by sulfate-reducing-bacteria by plastic で 示す MPBの Fig.2A), CFCs of SRB and MPB in stopped tube culture with medium containing sulfate(3.6 mm)as sulfur source; B), Methane formation and cell growth of methanogens versus time; C), The changes in conentration of sulfate and evolved sulfide in the medium versus time. The means of colony counts from duplicate roll tubes are plotted. は コ ロニ-はPhoto.1(平 コ ロニ ー はPhoto.2(平 板)と 板)で 示す 生 物質 を添 加 して い ない培 地 に よ る培 養 Fig,2(A), (B), (C)は 抗 生 物 質 を 添加 して い な い培 地 の 培養 結 果 を示 す SRBのCFCsは1日 /mlか cul- は の 希 釈 率 範 囲 で コ ロニ ー を形 成 させ た SRBの 1.抗 culti- petri dish タン菌 の 培養 液 をサ ンプ リン グ し, SRBに の 希 釈 率, MPBに 233 plastic petri dish of methane-producing-bacteria tivated 化 性 メ タン生 成 菌 の 共 生関 係 ら 個/mLま 間 で7.2 10E個 を め ぐ る競 合 は, ま ず 硫 酸 塩 が 存 在 す る と き にHzは で6倍 以 上 に 増 加 し た そ の 後, 時 間 の 経過 と共 に菌 数 が減 少 し, 最 終 的 にCFCs SBRに は 個/mLま これ は硫 化 物 が メ タン菌 の 生 育 に提 供 され るこ とに よ リメ タン菌 エ ネ ル ギ-を 獲 得 し, 初 期 の の 増 殖 及 び メタ ン生 成 を促 進 した と推定 され る そ して で 低 下 した(Fig.20A)) 酸 塩 の 還 元 に よ りSRBが 電 子供 与 体 と して 消 費 され, 次 い で生 成 した 硫 菌 の増 殖 を もた ら した もの と考 え られ る またFig-2(C) 硫 酸 塩 濃 度 の減 少 に よ りSRBの に 示 した よ うに硫 酸塩 濃度 の 減 少 に伴 い硫 酸 還 元菌 の 生 果 的 に メ タ ン菌 が 優 占種 と な る と考 え られ た SRBと 育 が制 限 され, 硫 酸 還 元菌 のCFCs減 MPB両 考 え られ る これ に対 し, MPBの 少 の原 因 に な る と 初 期 のCFCsの 菌 の コ ロニ ー の 計 数 結 果 は2オ ー ダー の 差 が あ り, SRBはMPBと 増殖 生 育 が 制 限 を受 け, 結 比 べ 僅 か で あ っ た の で, 測 定 し た は非常 に遅 く, 2日 目に対 数 増 殖期 に入 った 後, 2日 目 菌 の 乾 燥 重 量 は全 て メ タン菌 と見 な して もよい と考 え ら か ら の2日 間 個/mLか れ る こ れ は ま たFig..2(B)に ら 個/mLま で 急 速 に 増 殖 した つ ま り, メ タン 菌 と硫 酸 還 元 菌 のH2 示 す よ うに メ タ ン菌 体 密 度 の 経 時 変化 は メ タ ン菌 のCFCsの -41- 経 時 変化 と同 じ傾 向
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5 Fig. 3 CUFs of SRB and MPB in stopped tube culture with medium containing antibiotics and sulfate (3.6mM) as sulfur source; B), CH4 formation and cell growth of methanogens versus time; C), The changes in concentration of sulfate and evolved sulfide in the medium versus time. The means of colony counts from duplicate roll tubes are plotted. 2) M. Sai Ram, L. Singh and S. I. Alam (1993): Effect of sulfate and nitrate on anaerobic degradation of night soil. Bioresource Technology, 45: ) Zhen Ya Zhang and Takaaki Maekawa (1996): Effects of sulfur-containing compounds on the growth and methane production of acclimatedmixed methanogens. Biomass and Bioenergy, 10 (1): ) W. Jack Jones, W. B. Whitman, R. D. Fields and R. S. Wolfe (1983): Growth and Plating Efficiency of Methanococci on Agar medium, Appl. Environ. Microbiol., 46: ) Z. Isa, S. Grusenmeyer and W. Verstraete (1986: Sulfate-reduction relative to methane production in high-rate anaerobic digestion: technical aspects, Appl. Env. Microbiol., 51: ) P. H. Ronnow and Lars A. H. Gunnarsson (1982): Response of growth and methane production to limiting amounts of sulfide and ammonia in two thermophilic methanogenic bacteria, FFMS. Microbiol. Lett., 14, (1982)
6 The Symbiotic Relation Between Sulfate-Reducing-Bacteria and Methanogens in Methane Fermentation from CO2 and H2 Takaaki MAEKAWA and Zhen Ya ZHANG Institute of Agricultural and Forest Engineering University of Tsukuba, Tennodai, Ibaraki 305, Japan Abstract The competition for H2 substrate and the symbiotic relation between sulfate-reducing-bacteria (SRB) and methane-producing-bacteria (MPB) were investigated in this study for the acclimated-mixed-methanogens culture from CO2 and H2. The medium was cultured with antibiotics where growth of eubateria was inhibited but not methanogens, and without antibiotics where sulfate was used as a sulfur source. In the case of the medium without antibi- vation, while CFCs of MPB increased less rapidly. Entering growth phase after 2 days, CFCs of MPB rapidly in- inhibited, thus sulfate reduction to sulfide was impossible. Comparison of CFCs of MPB and methane gas formation showed that numbers were less in the medium containing antibiotics, than in the medium without antibiotics. MPB were not capable of reducing sulfate to sulfide, thus CFCs and methane gas formation were reduced. These results suggested that SRB, which exists symbiotically with predominant methanogens, produced a sulfur source by reducing sulfate to sulfide. The reduction consequentially provided an environment for the development of strictly anaerobic organisms, thereby accelerating the growth of methanogens. The role that SRB played in symbiotic methanogenesis was evaluated and examined in this study. Keywords: sulfate-reducing-bacteria, methane-producing-bacteria, symbiotic relation, sulfur source, sulfate reduction
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