Phylogenetic description of immobilized methanogenic community using real-time PCR in a fixed-bed anaerobic digester

Sawayama, Shigeki; Tsukahara, Kenichiro; Yagishita, Tatsuo

doi:10.1016/j.biortech.2005.02.011

Cited by 63 publications

(22 citation statements)

References 23 publications

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“…A number of studies have quantified methanogens belonging to different phylogenetic groups with techniques such as dot blot hybridization and fluorescent in situ hybridization (27,38,57), as well as qPCR (19,41,42,(54)(55)(56). Quantitation in these studies has targeted 16S rRNA or the rRNA gene, but studies by Springer et al (49) and Bapteste et al (3) suggest that the mcrA gene demonstrates the same phylogenetic relationships as the 16S rRNA gene between organisms.…”

Section: Discussionmentioning

confidence: 99%

“…Real-time quantitative PCR (qPCR) is an alternate technique capable of determining the copy number of a particular gene present in the DNA extracted from an environmental sample. Only a few studies have used qPCR to quantitatively examine different clades within methanogen communities, and most of these studies have exclusively targeted the 16S rRNA gene (19,41,42,(54)(55)(56). Far fewer researchers have used qPCR to quantify methanogen clades by targeting the mcrA (21,34,45), and these studies were limited to only a few phylogenetic groups.…”

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confidence: 99%

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mcrA -Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities

Steinberg

Regan

2009

Appl Environ Microbiol

234

138

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Methanogens are of great importance in carbon cycling and alternative energy production, but quantitation with culture-based methods is time-consuming and biased against methanogen groups that are difficult to cultivate in a laboratory. For these reasons, methanogens are typically studied through culture-independent molecular techniques. We developed a SYBR green I quantitative PCR (qPCR) assay to quantify total numbers of methyl coenzyme M reductase ␣-subunit (mcrA) genes. TaqMan probes were also designed to target nine different phylogenetic groups of methanogens in qPCR assays. Total mcrA and mcrA levels of different methanogen phylogenetic groups were determined from six samples: four samples from anaerobic digesters used to treat either primarily cow or pig manure and two aliquots from an acidic peat sample stored at 4°C or 20°C. Only members of the Methanosaetaceae, Methanosarcina, Methanobacteriaceae, and Methanocorpusculaceae and Fen cluster were detected in the environmental samples. The three samples obtained from cow manure digesters were dominated by members of the genus Methanosarcina, whereas the sample from the pig manure digester contained detectable levels of only members of the Methanobacteriaceae. The acidic peat samples were dominated by both Methanosarcina spp. and members of the Fen cluster. In two of the manure digester samples only one methanogen group was detected, but in both of the acidic peat samples and two of the manure digester samples, multiple methanogen groups were detected. The TaqMan qPCR assays were successfully able to determine the environmental abundance of different phylogenetic groups of methanogens, including several groups with few or no cultivated members.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

mcrA -Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities

Steinberg

Regan

2009

Appl Environ Microbiol

234

138

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“…x 10 9 cells/mL vs. 1.53 x 10 8 cells/mL, respectively) (Sawayama et al 2006;Yang et al 2004). Thus, fixed-bed biofilm reactors allow for MA numbers comparable or even greater than BAC numbers.…”

Section: Quantification Of Reactor Samplesmentioning

confidence: 96%

Effect of Sulfide Inhibition and Organic Shock Loading on Anaerobic Biofilm Reactors Treating a Low‐Temperature, High‐Sulfate Wastewater

McDonald¹,

Parkin²

2009

Water Environment Research

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McDonald, Heather Brown. "The effect of sulfide inhibition and organic shock loading on anaerobic biofilm reactors treating a lowtemperature, high-sulfate wastewater. mg COD/L. It was determined that long-term stability could be achieved in reactors that contained well balanced, stable populations of lactate-and propionate-degrading SRB and aceticlastic methanogens. Significant populations of fermenters present resulted in an imbalance which caused lactate to be routed through an additional pathway where propionate was formed. Greater numbers of MA than bacteria were found in all reactors.This may be attributed to the availability of acetate in the reactors for MA consumption and to using the immobilized fixed bed reactor type. Aceticlastic methanogens were the dominant methanogen, and were observed to remove nearly all acetate produced in all reactors. SRB were observed to remove lactate in microbially balanced reactors, whereas fermenters degraded lactate in reactors with less balanced populations.

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“…Therefore, the activity of the hydrogenotrophic methanogens are crucial for a stable and efficient process performance. The performance and activity of hydrogenotrophic methanogens in anaerobic conversion of simplesoluble type of substrates (such as acetate, ethanol, methanol, glucose and propionate) and various types of wastewaters (such as olive mill) have been covered and discussed in recent research activities selected here (Gijzen et al 2000;Imachi et al 2000;Gonzalez-Gil et al 2001;Syutsubo et al 2001;Delbes et al 2001;Paulo et al 2002;Paulo et al 2003;McHugh et al 2004;Bertin et al 2004;Sipma et al 2004;Tada et al 2005;Tang et al 2005;Roest et al 2005;Sawayama et al 2006;Shigematsu et al 2006). On the contrary, the role and activity of hydrogenotrophic methanogens in anaerobic conversion of biomass/complex organic materials to methane has not been regularly discussed, and there exists relatively less amount of data during the previous decade (Blotevogel et al 1985;Jarvis et al 1995;Zhu et al 1997;Schnürer et al 1999).…”

Section: The Role Of Hydrogenotrophic Methanogensmentioning

confidence: 98%

The roles of acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of biomass to methane: a review

Demirel

Scherer

2008

Rev Environ Sci Biotechnol

967

497

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Among different conversion processes for biomass, biological anaerobic digestion is one of the most economic ways to produce biogas from various biomass substrates. In addition to hydrolysis of polymeric substances, the activity and performance of the methanogenic bacteria is of paramount importance during methanogenesis. The aim of this paper is primarily to review the recent literature about the occurrence of both acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of particulate biomass to methane (not wastewater treatment), while this review does not cover the activity of the acetate oxidizing bacteria. Both acetotrophic and hydrogenotrophic methanogens are essential for the last step of methanogenesis, but the reports about their roles during this phase of the process are very limited. Despite, some conclusions can still be drawn. At low concentrations of acetate, normally filamentous Methanosaeta species dominate, e.g., often observed in sewage sludge. Apparently, high concentrations of toxic ionic agents, like ammonia, hydrogen sulfide (H 2 S) and volatile fatty acids (VFA), inhibit preferably Methanosaetaceae and especially allow the growth of Methanosarcina species consisting of irregular cell clumps, e.g., in cattle manure. Thermophilic conditions can favour rod like or coccoid hydrogenotrophic methanogens. Thermophilic Methanosarcina species were also observed, but not thermophilic Methanosaetae. Other environmental factors could favour hydrogentrophic bacteria, e.g., short or low retention times in a biomass reactor. However, no general rules regarding process parameters could be derivated at the moment, which favours hydrogenotrophic methanogens. Presumably, it depends only on the hydrogen concentration, which is generally not mentioned in the literature.

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Phylogenetic description of immobilized methanogenic community using real-time PCR in a fixed-bed anaerobic digester

Cited by 63 publications

References 23 publications

mcrA -Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities

mcrA -Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities

Effect of Sulfide Inhibition and Organic Shock Loading on Anaerobic Biofilm Reactors Treating a Low‐Temperature, High‐Sulfate Wastewater

The roles of acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of biomass to methane: a review

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