Šimůnek Jiří, Killer Jiří, Sechovcová Hana, Šimůnek Jiří, Pechar Radko, Rada Vojtěch, Švec Pavel, Sedláček Ivo
Institute of Animal Physiology and Genetics v.v.i. of the Czech Academy of Sciences, Vídeňská 1083, 142 20, Prague4, Krč, Czech Republic.
Faculty of Agrobiology, Food and Natural Resources, Department of Microbiology, Nutrition and Dietetics, Czech University of Life Sciences, Kamýcká 129, Prague 6, 165 00, Suchdol, Czech Republic.
Folia Microbiol (Praha). 2018 May;63(3):391-399. doi: 10.1007/s12223-017-0577-9. Epub 2017 Dec 21.
Gram-stain-positive, catalase and oxidase-negative and short rod-shaped bacterium C10 with occasional branching was isolated under strictly anaerobic conditions from the rumen fluid of a red deer (Cervus elaphus) in the course of study attempting to uncover new xylanolytic and cellulolytic rumen bacteria inhabiting the digestive tract of wild ruminants in the Czech Republic. The anaerobic M10 medium containing bovine rumen fluid and carboxymethylcellulose as a defined source of organic carbon was used in the process of bacterial isolation. The 16S rRNA gene similarity revealed recently characterized new species Actinomyces succiniciruminis Am4 (GenBank accession number of the gene retrieved from the complete genome: LK995506) and Actinomyces glycerinitolerans G10 (GenBank accession number from the complete genome: NZFQTT01000017) as the closest relatives (99.7 and 99.6% gene pairwise identity, respectively), followed by the Actinomyces ruminicola DSM 27982 (97.2%, in all compared fragment of 41468 pb). Due to the taxonomic affinity of the examined strain to both species A. succiniciruminis and A. glycerinitolerans, its taxonomic status towards these species was evaluated using variable regions of rpsA (length of 519 bp) and rplB (597 bp) gene sequences amplified based on specific primers designed so as to be applicable in differentiation, classification, and phylogeny of Actinomyces species/strains. Comparative analyses using rpsA and rplB showed 98.5 and 97.9% similarities of C10 to A. succiniciruminis, respectively, and 97.5 and 97.6% similarities to A. glycerinitolerans, respectively. Thus, gene identities revealed that the evaluated isolate C10 (=DSM 100236 = LMG 28777) is a little more related to the species A. succiniciruminis isolated from the rumen of a Holstein-Friesian cow than A. glycerinitolerans. Phylogenetic analyses confirmed affinity of strain C10 to both recently characterized species. Unfortunately, they did not allow the bacterial strain to be classified into a particular species. Phenotypic characterization suggested similar conclusions. This brief contribution is aimed at classification and detailed phenotypic characterization of bacterial strain C10 isolated from the rumen of a wild red deer exhibiting, from the point of view of Actinomyces species, noteworthy cellulolytic and xylanolytic activities.
在试图发现捷克共和国野生反刍动物消化道中新型木聚糖分解和纤维素分解瘤胃细菌的研究过程中,在严格厌氧条件下从一只马鹿(Cervus elaphus)的瘤胃液中分离出革兰氏染色阳性、过氧化氢酶和氧化酶阴性且呈短杆状、偶尔有分支的细菌C10。在细菌分离过程中使用了含有牛瘤胃液和羧甲基纤维素作为有机碳特定来源的厌氧M10培养基。16S rRNA基因相似性显示,最近鉴定的新物种琥珀酸瘤胃放线菌Am4(从完整基因组检索到的该基因的GenBank登录号:LK995506)和甘油耐受放线菌G10(来自完整基因组的GenBank登录号:NZFQTT01000017)是最接近的亲缘种(基因两两同一性分别为99.7%和99.6%),其次是瘤胃放线菌DSM 27982(97.2%,在41468 pb的所有比较片段中)。由于所检测菌株与琥珀酸瘤胃放线菌和甘油耐受放线菌这两个物种在分类学上具有亲缘关系,因此基于设计用于放线菌物种/菌株分化、分类和系统发育的特异性引物,扩增rpsA(长度为519 bp)和rplB(597 bp)基因序列的可变区,以评估其与这些物种的分类地位。使用rpsA和rplB进行的比较分析表明,C10与琥珀酸瘤胃放线菌的相似性分别为98.5%和97.9%,与甘油耐受放线菌的相似性分别为97.5%和97.6%。因此,基因同一性表明,所评估的分离株C10(=DSM 100236 = LMG 28777)与从荷斯坦 - 弗里生奶牛瘤胃中分离出的琥珀酸瘤胃放线菌的亲缘关系比与甘油耐受放线菌的亲缘关系稍近。系统发育分析证实了菌株C10与这两个最近鉴定的物种的亲缘关系。不幸的是,它们无法将该细菌菌株归入特定物种。表型特征分析也得出了类似结论。本简短报告旨在对从野生马鹿瘤胃中分离出的细菌菌株C10进行分类和详细的表型特征分析,从放线菌物种的角度来看,该菌株具有值得注意的纤维素分解和木聚糖分解活性。