多重多重 PCR 检测在 4 °C 真空保存的新鲜希腊乳清干酪中占主导地位的非典型中肠白色念珠菌亚种 jonggajibkimchii 表型的末端腐败微生物关联

Nikoletta Sameli, E. Sioziou, L. Bosnea, S. Paramithiotis, J. Samelis
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引用次数: 0

摘要

将物种特异性多重 PCR 方法和表型测试结合起来,评估了 24 个具有代表性的中肠白色念珠菌不同分离物之间的生化和基因型差异,这些分离物以前曾在 4 °C 下储存 40 天的变质真空包装 Anthotyros 乳清奶酪中发现并通过 16S rRNA 基因测序鉴定。根据其表型(API 50 CHL)特征,这 24 个分离物包括 6 个多菌株生物型和 7 个单菌株生物型。只有两个单菌株生物型(L4A 和 L4B)从蔗糖中产生粘液(葡聚糖),只有四个生物型(L2A-L2C、L3;7 个分离株)发酵 L-阿拉伯糖;其余 15 个分离株(生物型 L1A-L1F)是葡聚糖阴性、低聚发酵的 Ln. mesenteroides 变异株,能发酵 D-木糖并在 37 ℃ 下生长。根据其多重 PCR(rpoB、araA、dsr 和 sorA)基因图谱与四种 Ln. mesenteroides subsp.在与 cremoris(rpoB)、dextranicum(rpoB/dsr)、mesenteroides(rpoB/araA/dsr/sorA)和 jonggajibkimchii(rpoB/araA/dsr)四个 Ln mesenteroides 亚种的模式菌株进行比较后,没有一个分离株被归入前两个亚种,只有四个分离株(L2A 和 L2C)被归入 mesenteroides 亚种。十个分离物具有琼加吉布金氏亚种的特征,而其他十个分离物具有第五个非典型特征(rpoB/dsr/sorA),似乎更接近右旋糖酐亚种。尤其是非典型生物型 L1B,代表了在 Anthotyros 乳清干酪腐败中最普遍的精神滋养型 Ln. mesenteroides 亚种 jonggajibkimchii(rpoB/araA/dsr)基因型,值得进一步进行生化和分子特征研究。
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Multiplex-PCR Detection of an Atypical Leuconostoc mesenteroides subsp. jonggajibkimchii Phenotype Dominating the Terminal Spoilage Microbial Association of a Fresh Greek Whey Cheese Stored at 4 °C in Vacuum
A species-specific multiplex-PCR method and phenotypic tests were combined to evaluate biochemical and genotypic differences between 24 representative Leuconostoc mesenteroides diverse isolates previously found to dominate in spoiled, vacuum-packed Anthotyros whey cheeses stored at 4 °C for 40 days and identified by 16S rRNA gene sequencing. Based on their phenotypic (API 50 CHL) profiles, the 24 isolates comprised 6 multi-strain and 7 single-strain biotypes. Only two single-strain biotypes (L4A and L4B) produced slime (dextran) from sucrose, and only four biotypes (L2A–L2C, L3; 7 isolates) fermented L-arabinose; the remaining 15 isolates (biotypes L1A–L1F) were dextran-negative, oligofermenting Ln. mesenteroides variants, able to ferment D-xylose and grow at 37 °C. Based on their multiplex-PCR (rpoB, araA, dsr, and sorA) gene profiles in comparison with those of the type strains of the four Ln. mesenteroides subsp. cremoris (rpoB), dextranicum (rpoB/dsr), mesenteroides (rpoB/araA/dsr/sorA), and jonggajibkimchii (rpoB/araA/dsr), no isolate was assigned to the first two subspecies and only four isolates (L2A and L2C) to the subsp. mesenteroides. Ten isolates shared the subsp. jonggajibkimchii profile, while the other ten ones have a fifth atypical profile (rpoB/dsr/sorA), seemingly being closer to the subsp. dextranicum. Particularly the atypical biotype L1B representatives of the most prevalent psychrotrophic Ln. mesenteroides subsp. jonggajibkimchii (rpoB/araA/dsr) genotype at Anthotyros whey cheese spoilage deserve further biochemical and molecular characterization studies.
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