Pechar Radko, Killer Jiří, Mekadim Chahrazed, Geigerová Martina, Rada Vojtěch
Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences, Kamýcká 129, Suchdol, 165 00, Prague 6, Czech Republic.
Food Research Institute Prague, Radiová 1285/7, Hostivař, 102 00, Prague 10, Czech Republic.
Curr Microbiol. 2017 Nov;74(11):1324-1331. doi: 10.1007/s00284-017-1320-0. Epub 2017 Jul 29.
Occurrence of bifidobacteria, known as health-promoting probiotic microorganisms, in the digestive tract of wild pigs (Sus scrofa) has not been examined yet. One hundred forty-nine fructose-6-phosphate phosphoketolase positive bacterial strains were isolated from colonic content of twenty-two individuals of wild pigs originated from four localities in the Czechia. Based on PCR-DGGE technique targeting the variable V3 region of the 16S rRNA genes, strains were initially differentiated into four groups represented by: (i) probably a new Bifidobacterium species (89 strains), (ii) B. boum/B. thermophilum/B. thermacidophilum subsp. porcinum/B. thermacidophilum subsp. thermacidophilum (sub)species (49 strains), (iii) Pseudoscardovia suis (7 strains), and (iv) B. pseudolongum subsp. globosum/B. pseudolongum subsp. pseudolongum (4 strains), respectively. Given the fact that DGGE technique did not allow to differentiate the representatives of thermophilic bifidobacteria and B. pseudolongum subspecies, strains were further classified by the 16S rRNA and thrS gene sequences. Primers targeting the variable regions of the latter gene were designed to be applicable in identification and phylogeny of Bifidobacteriaceae family. The 16S rRNA-derived phylogenetic study classified members of the first group into five subgroups in a separated cluster of thermophilic bifidobacteria. Comparable results were obtained by the thrS-derived phylogenetic analysis. Remarkably, variability among thrS sequences was higher compared with 16S rRNA gene sequences. Overall, molecular genetic techniques application allowed to identify a new Bifidobacterium phylotype which is predominant in the digestive tract of examined wild pigs.
双歧杆菌作为具有促进健康作用的益生菌微生物，其在野猪（Sus scrofa）消化道中的存在情况尚未得到研究。从来自捷克四个地区的22头野猪的结肠内容物中分离出149株6-磷酸果糖磷酸酮醇酶阳性细菌菌株。基于针对16S rRNA基因可变V3区域的PCR-DGGE技术，这些菌株最初被分为四组，分别为：（i）可能是一个新的双歧杆菌物种（89株），（ii）猪双歧杆菌/嗜热双歧杆菌/嗜热嗜酸双歧杆菌猪亚种/嗜热嗜酸双歧杆菌嗜热嗜酸亚种（亚）种（49株），（iii）猪假斯卡多维亚菌（7株），以及（iv）假长双歧杆菌球形亚种/假长双歧杆菌假长亚种（4株）。鉴于DGGE技术无法区分嗜热双歧杆菌和假长双歧杆菌亚种的代表菌株，因此通过16S rRNA和thrS基因序列对这些菌株进行了进一步分类。针对后一个基因可变区域设计的引物适用于双歧杆菌科的鉴定和系统发育研究。基于16S rRNA的系统发育研究将第一组的成员在嗜热双歧杆菌的一个单独聚类中分为五个亚组。通过基于thrS的系统发育分析也获得了类似的结果。值得注意的是，与16S rRNA基因序列相比，thrS序列之间的变异性更高。总体而言，分子遗传学技术的应用使得能够鉴定出一种在被检查野猪消化道中占主导地位的新双歧杆菌系统型。
