Studies about Bacteria Quantification using Molecular Biological Method and Mechanism of Biogas Production.

甲烷和氢气的浓度测量使用半导体气体监测系统（mBA-3000）提供的医疗用途。甲烷和氢的浓度之间没有相关性，甲烷和一氧化二氮的浓度之间也没有相关性。甲烷和氨浓度之间存在一周的正相关关系。我们发明了一种利用半导体气体传感器监测反刍动物呼出气体的甲烷和氢气监测仪（mBA-5000）。该监测器可以连续20小时每10秒检测一次甲烷（1 ~ 1000ppm）和氢气（1 ~ 100ppm）的浓度，存储的数据可以通过串口转换到个人电脑。mBA-5000被放置在荷斯坦奶牛喂养的代谢室中。采食后甲烷气体浓度逐渐升高，在采食后1小时左右达到峰值，全天保持在600 ~ 1000ppm。在喂食后5 ~ 15分钟观察到产氢量，在喂食后30分钟达到峰值。加料后150分钟，产氢量恢复到0 ppm，并一直保持到下一次加料时为0 ppm。由此推测，饲喂后产生的氢气可能是瘤胃发酵产生的氢气或质子溢出。不同的动物产生甲烷的能力应该是不同的，实际上氢气的峰值水平从20到90 ppm不等，这取决于实验中使用的牛。利用16S核糖体RNA基因和甲基辅酶M还原酶基因序列构建的引物对产甲烷菌进行PCR检测和定量。PCR分析结果表明，灭羊瘤胃产甲烷菌与未灭羊瘤胃产甲烷菌有显著差异。

The concentrations of methane and hydrogen gas were measured using a semiconductor gas monitor system (mBA-3000) provided for medical use. There was no correlation between the methane and hydrogen concentrations, or between methane and nitrous oxide concentrations. There was a week positive correlation between the methane and ammonia concentrations. We invented a methane and hydrogen gas monitor (mBA-5000) for the expired breath of ruminant animals using semiconductor gas sensor. This monitor can detect the concentration of methane (1 to 1000 ppm) and hydrogen (1 to 100 ppm) every 10 seconds for 20 hours, and the stored data can convert to a personal computer via serial port. The mBA-5000 was placed in the metabolic chamber in which a Holstein cow was feeding. The concentrations of methane gas were increased after feeding and peaked at around 1 hour after feeding, and were keeping 600 to 1000 ppm all day long. The hydrogen gas production was observed from 5 to 15 minutes after feeding, and peaked at 30 minutes after feeding. The hydrogen production returned to 0 ppm until 150 minutes after feeding, and had been keeping 0 ppm until the next feeding. This observation reads to a speculation that the hydrogen gas production after feeding might be the overflow of hydrogen or proton produced by the rumen fermentation. The capacity of methane producing rate should be different between the animals, and actually the peak levels of hydrogen gas were varied from 20 to 90 ppm depending on the cattle used in this experiment. PCR detection and quantification of methanogen was provided using the primers constructed from the sequences of 16S ribosomal RNA gene and methyl coenzyme M reductase gene. The results of PCR analysis indicated that the methanogens of the faunated rumen differed significantly from those of the unfaunated rumen.