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Structure and Topology of Microbial Communities in the Major Gut Compartments of Melolontha melolontha Larvae (Coleoptera: Scarabaeidae)

机译:Melolontha melolontha幼虫(鞘翅目:Scarabaeidae)主要肠道区的微生物群落结构和拓扑。

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摘要

Physicochemical gut conditions and the composition and topology of the intestinal microbiota in the major gut compartments of the root-feeding larva of the European cockchafer (Melolontha melolontha) were studied. Axial and radial profiles of pH, O2, H2, and redox potential were measured with microsensors. Terminal restriction fragment length polymorphism (T-RFLP) analysis of bacterial 16S rRNA genes in midgut samples of individual larvae revealed a simple but variable and probably nonspecific community structure. In contrast, the T-RFLP profiles of the hindgut samples were more diverse but highly similar, especially in the wall fraction, indicating the presence of a gut-specific community involved in digestion. While high acetate concentrations in the midgut and hindgut (34 and 15 mM) corroborated the presence of microbial fermentation in both compartments, methanogenesis was confined to the hindgut. Methanobrevibacter spp. were the only methanogens detected and were restricted to this compartment. Bacterial 16S rRNA gene clone libraries of the hindgut were dominated by clones related to the Clostridiales. Clones related to the Actinobacteria, Bacillales, Lactobacillales, and γ-Proteobacteria were restricted to the lumen, whereas clones related to the β- and δ-Proteobacteria were found only on the hindgut wall. Results of PCR-based analyses and fluorescence in situ hybridization of whole cells with group-specific oligonucleotide probes documented that Desulfovibrio-related bacteria comprise 10 to 15% of the bacterial community at the hindgut wall. The restriction of the sulfate-reducer-specific adenosine-5′-phosphosulfate reductase gene apsA to DNA extracts of the hindgut wall in larvae from four other populations in Europe suggested that sulfate reducers generally colonize this habitat.
机译:研究了欧洲金龟子(Melolontha melolontha)的根食幼虫主要肠道区室的理化肠道条件以及肠道菌群的组成和拓扑。用微传感器测量pH,O2,H2和氧化还原电势的轴向和径向轮廓。个体幼虫中肠样品中细菌16S rRNA基因的末端限制性片段长度多态性(T-RFLP)分析显示一个简单但可变且可能非特异性的群落结构。相比之下,后肠样品的T-RFLP图谱则更加多样,但高度相似,尤其是在壁部分,这表明存在消化相关的肠道特异性群落。尽管中肠和后肠中醋酸盐的浓度较高(分别为34和15 mM)证实了两个隔室中都存在微生物发酵,但甲烷生成仅限于后肠。甲烷短杆菌属。是唯一检测到的产甲烷菌,并仅限于该区域。后肠的细菌16S rRNA基因克隆文库由与梭菌相关的克隆所控制。与放线菌,芽孢杆菌,乳杆菌和γ-变形杆菌有关的克隆仅限于管腔,而与β-和δ-变形细菌有关的克隆仅在后肠壁上发现。基于PCR的分析结果和全细胞与组特异性寡核苷酸探针的荧光原位杂交结果表明,脱硫弧菌相关细菌占后肠壁细菌群落的10%至15%。硫酸盐还原剂特异性腺苷5'-磷酸硫酸盐还原酶基因apsA对欧洲其他四个种群的幼虫后肠壁DNA提取物的限制表明,硫酸盐还原剂通常在该栖息地定居。

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