Aquilanti Lucia, Santarelli Sara, Silvestri Gloria, Osimani Andrea, Petruzzelli Annalisa, Clementi Francesca
Department of Food Science, Polytechnic University of Marche, via Brecce Bianche, 60131 Ancona, Italy.
Int J Food Microbiol. 2007 Nov 30;120(1-2):136-45. doi: 10.1016/j.ijfoodmicro.2007.06.010. Epub 2007 Jun 13.
We have investigated the bacteria and yeast ecology of the typical Italian Ciauscolo salami that is produced in Central Italy using a polyphasic approach based on culture-dependent and -independent methods. The physico-chemical analyses showed a progressive drop in pH and water activity (aw) during ripening. The viable counts revealed a dominance of lactic acid bacteria (LAB) over coagulase negative cocci (CNC) and yeasts. From the molecular identification of the isolates, the prevalence of Lactobacillus curvatus, Lb. plantarum and Staphylococcus xylosus was shown among the bacteria, while Debaryomyces hansenii was the prevalent species among the yeasts, and it was isolated throughout the whole ripening process. Minority species, namely Rhodotorula mucillaginosa and Trichosporon brassicae, were also recovered from the meat batter. The total microbial community was profiled without cultivation by analyzing the DNA that was directly extracted from the salami samples. Moreover, the cultivable community was profiled by analyzing the DNA recovered from bulk cells that were obtained by harvesting the colonies from serial-dilution agar plates. The 16S rRNA gene V1 and V3 regions were used as targets in the denaturing gradient gel electrophoresis (DGGE) profiling of the LAB and CNC communities, respectively, while the diversity and dynamics of the yeast population were assessed by analyzing a portion of the 28S rRNA gene. Our findings suggest that the microbial diversity of fermented meat products can be successfully investigated by this polyphasic approach that is based on the assessment of both the total and the cultivable community diversity.
我们采用基于依赖培养法和非依赖培养法的多相方法,对意大利中部生产的典型意大利恰乌斯科洛香肠的细菌和酵母生态进行了研究。理化分析表明,在成熟过程中pH值和水分活度(aw)逐渐下降。活菌计数显示,乳酸菌(LAB)比凝固酶阴性球菌(CNC)和酵母占优势。通过对分离株的分子鉴定,弯曲乳杆菌、植物乳杆菌和木糖葡萄球菌在细菌中占优势,而汉逊德巴利酵母是酵母中的优势种,并且在整个成熟过程中都能分离到。少数菌种,即粘红酵母和油菜毛孢子菌,也从肉馅中分离出来。通过分析直接从香肠样品中提取的DNA,对未培养的总微生物群落进行了分析。此外,通过分析从连续稀释琼脂平板上收获的菌落获得的大量细胞中回收的DNA,对可培养群落进行了分析。在乳酸菌和凝固酶阴性球菌群落的变性梯度凝胶电泳(DGGE)分析中,分别将16S rRNA基因的V1和V3区域作为靶标,而通过分析28S rRNA基因的一部分来评估酵母种群的多样性和动态。我们的研究结果表明,通过这种基于总群落和可培养群落多样性评估的多相方法,可以成功地研究发酵肉制品的微生物多样性。