Archaeal microbiota population in piglet feces shifts in response to weaning: Methanobrevibacter smithii is replaced with Methanobrevibacter boviskoreani
Archaeal microbiota population in piglet feces shifts in response to weaning: Methanobrevibacter smithii is replaced with Methanobrevibacter boviskoreani
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DOI：
10.1093/femsle/fnv064
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发表时间：
2015-05-01
影响因子：
2.1
通讯作者：
Callegari, Maria Luisa
中科院分区：
文献类型：
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作者：
Federici, Sara;Miragoli, Francesco;Callegari, Maria Luisa
产甲烷菌通常存在于猪的肠道中。我们从9头仔猪的粪便中提取DNA，对肠道古菌种群进行了分析。我们进行了针对16S rRNA基因V6-V8区域的聚合酶链式反应。随后的变性梯度凝胶电泳(DGGE)发现了以前在猪身上未发现的牛博维斯克雷亚氏甲烷短杆菌。我们用MCRA基因的PCR-DGGE分析和随后的测序证实了这些数据。在63日龄时，粪便样本中唯一的条带对应于Boviskoreani分枝杆菌。DGGE分析还表明，28d时数量较多的史密斯甲烷短杆菌在42d时数量急剧减少，63d时完全消失。为了验证这些数据，我们用定量聚合酶链式反应(QPCR)对斯氏分枝杆菌和古菌总数进行了量化；此外，我们还根据博维斯科雷尼分枝杆菌的16S rRNA基因设计了一套新的物种特异性引物。定量聚合酶链式反应结果证实，随着时间的推移，史密斯分枝杆菌的数量减少，而博维斯克雷亚氏分枝杆菌的数量同时增加。在63日龄时，古生菌和博维斯科雷亚氏支原体的基因组总数基本相当，说明博维斯科雷亚氏支原体是优势古生菌。这项研究表明，在断奶过程中，古生菌种群发生了转移，博维斯克里亚尼支原体取代了斯米蒂亚氏支原体。
Methanogens commonly inhabit swine intestine. We analyzed the gut archaeal population by extracting DNA from the feces of nine piglets. We performed PCR to target the V6-V8 region of the 16S rRNA gene. Subsequent denaturing gradient gel electrophoresis (DGGE) revealed the presence of Methanobrevibacter boviskoreani, which has not previously been identified in pigs. We confirmed these data with a PCR-DGGE analysis of the mcrA gene, and subsequent sequencing. At 63 days old, the only band in fecal samples corresponded to M. boviskoreani. The DGGE analysis also showed that Methanobrevibacter smithii, which was abundant at 28 days, was dramatically reduced at 42 days, and it completely disappeared at 63 days. To confirm these data, we quantified M. smithii and the total archaeal population by quantitative PCR (qPCR); moreover, we designed a new set of species-specific primers based on the 16S rRNA gene of M. boviskoreani. The qPCR results confirmed the reduction in M. smithii over time and a simultaneous increase in M. boviskoreani. At 63 days, the total numbers of archaea and M. boviskoreani genomes were comparable, which suggested that M. boviskoreani represented the dominant archaea. This work showed that the archaeal population shifted during weaning, and M. boviskoreani replaced M. smithii.