Cook Kimberly L, Whitehead Terence R, Spence Cheryl, Cotta Michael A
AWMRU, USDA-ARS, Bowling Green, KY 42104, USA.
Anaerobe. 2008 Jun;14(3):172-80. doi: 10.1016/j.anaerobe.2008.03.003. Epub 2008 Mar 28.
Hydrogen sulfide, produced by sulfate-reducing bacteria (SRB), is one of the most potent malodors emitted from anaerobic swine waste storage systems. However, little is known about the prevalence and diversity of SRB in those systems. The goals of this study were to evaluate the SRB population in swine manure storage systems and to develop quantitative, real-time PCR (QRT-PCR) assays to target four of the SRB groups. Dissimilatory sulfite reductase (DSR) gene sequences were obtained from swine slurry stored in underground pits (43 clones) or in lagoons (34 clones). QRT-PCR assays were designed to target the dsrA gene of four novel groups of SRB. Sequences of dsrA clones from slurry samples grouped with those from three different cultured SRB: Desulfobulbus sp. (46 clones), Desulfovibrio sp. (24 clones and 5 isolates), and Desulfobacterium sp. (7 clones). However, DsrA sequences from swine slurry clones were generally less than 85% similar to those of cultured organisms. SRB from all four targeted SRB groups were detected in underground waste storage pits (6.6 x 10(3)-8.5 x 10(7) dsrA copies mL(-1) slurry), while only two groups of SRB were detected in lagoons (3.2 x 10(5)-2.5 x 10(6) dsrA copies mL(-1) slurry). To date, this is the only study to evaluate the phylogeny and concentration of SRB in any livestock waste storage system. The new QRT-PCR assays should facilitate sensitive, specific detection of the four novel groups of SRB in livestock waste storage systems.
由硫酸盐还原菌(SRB)产生的硫化氢是厌氧猪粪便储存系统中排放的最强烈的恶臭气味之一。然而,对于这些系统中SRB的流行情况和多样性知之甚少。本研究的目的是评估猪粪便储存系统中的SRB种群,并开发针对四个SRB组的定量实时PCR(QRT-PCR)检测方法。从储存在地下坑(43个克隆)或泻湖(34个克隆)中的猪粪浆中获得异化亚硫酸盐还原酶(DSR)基因序列。设计QRT-PCR检测方法以靶向四个新型SRB组的dsrA基因。来自粪浆样品的dsrA克隆序列与来自三种不同培养的SRB的序列分组在一起:脱硫球茎菌属(46个克隆)、脱硫弧菌属(24个克隆和5个分离株)和脱硫杆菌属(7个克隆)。然而,来自猪粪浆克隆的DsrA序列通常与培养生物的序列相似度低于85%。在地下废物储存坑中检测到了来自所有四个目标SRB组的SRB(6.6×10³-8.5×10⁷个dsrA拷贝 mL⁻¹粪浆),而在泻湖中仅检测到两组SRB(3.2×10⁵-2.5×10⁶个dsrA拷贝 mL⁻¹粪浆)。迄今为止,这是唯一一项评估任何家畜粪便储存系统中SRB系统发育和浓度的研究。新的QRT-PCR检测方法应有助于灵敏、特异检测家畜粪便储存系统中的四个新型SRB组。