Bacterial and fungal population dynamics in cider for distillation have so far been explored by culture-dependant methods. Cider for distillation can be produced by the spontaneous fermentation of apples that do not undergo any intervention during the process. In this study, cider microbiomes extracted from six tanks containing ciders for distillation from four producers in Normandy were characterized at three main stages of the fermentation process: fermentation Initiation (I), end of the alcoholic Fermentation (F) and end of the Maturation period (M). Cider samples were subjected to Illumina MiSeq sequencing (rRNA 16S V1-V3 and ITS1 region targeting) to determine bacterial and fungal communities. Yeasts (YGC), Zymomonas (mZPP) and lactic acid bacteria selective media (mMRS, mMLO, mPSM) were also used to collect 807 isolates. Alcoholic levels, glycerol, sugar content (glucose, fructose and sucrose), pH, total and volatile acidity, nitrogen, malic and lactic acid contents were determined at all sampling points. Alpha diversity indexes show significant differences (p < 0.05) in microbial populations between I, F and M. Fungal communities were characterized by microorganisms from the environment and phytopathogens at I followed by the association of yearsts with alcoholic fermentation like Saccharomyces and non-Saccharomyces yeasts (Hanseniaspora, Candida). A maturation period for cider leads to an increase of the Dekkera/Brettanomyces population, which is responsible for off-flavors in cider for all producers. Among bacterial communities, the genera community associated to malolactic fermentation (Lactobacillus sp., Leuconostoc sp., Oenococcus sp.) was the most abundant at F and M. Acetic acid bacteria such as Acetobacter sp., Komagataeibacter sp. and Gluconobacter sp. were also detected during the process. Significant differences (p < 0.05) were found in fungal and bacterial populations between the four producers and during the fermentation process. The development of microorganisms associated with cider spoilage such as Zymomonas mobilis, Lactobacillus collinoides or Brettanomyces/Dekkera sp. was anticipated by a metagenomic approach. The monitoring of microbial diversity via high throughput sequencing combined with physical-chemical analysis is an interesting approach to improve the fermentation performance of cider for distillation and therefore, the quality of Calvados.

迄今为止，已经通过依赖于文化的方法探索了用于蒸馏的苹果酒中细菌和真菌的种群动态。苹果酒的自然发酵可以产生蒸馏用的苹果酒，在此过程中无需进行任何干预。在这项研究中，从诺曼底的四个生产商的六个装有苹果酒的罐子中提取的苹果酒微生物群，在发酵过程的三个主要阶段进行了表征：发酵开始（I），酒精发酵结束（F）和成熟期结束（M）。对苹果酒样品进行Illumina MiSeq测序（rRNA 16S V1-V3和ITS1区域定位）以确定细菌和真菌群落。酵母（YGC），发酵单胞菌（mZPP）和乳酸菌选择性培养基（mMRS，mMLO，mPSM）也用于收集807种分离物。在所有采样点测定酒精含量，甘油，糖含量（葡萄糖，果糖和蔗糖），pH，总酸度和挥发性酸度，氮，苹果酸和乳酸含量。α多样性指数显示了I，F和M之间微生物种群之间的显着差异（p <0.05）。真菌群落的特征是来自环境的微生物和I处的植物病原体，其次是多年生与酒精发酵，如酿酒酵母和非酿酒酵母（Hanseniaspora，念珠菌）。苹果酒的成熟期导致Dekkera /酒仙子种群，对所有生产者的苹果酒产生异味。在细菌群落中，与苹果酸乳酸发酵相关的属群落（乳酸杆菌属，Leuconostoc属菌，Oenococcus属）在F和M处最丰富。醋酸菌，例如醋杆菌属，Komagataeibacter属。和葡糖杆菌属。在此过程中也被检测到。在四个生产者之间以及在发酵过程中，在真菌和细菌种群中发现了显着差异（p <0.05）。与苹果酒变质有关的微生物的发展，例如运动发酵单胞菌（Zymomonas mobilis），Collinoides乳杆菌或Brettanomyces / Dekkera sp。是宏基因组学方法所预期的。通过高通量测序与物理化学分析相结合的微生物多样性监测是一种有趣的方法，可以提高苹果酒蒸馏的发酵性能，从而改善Calvados的品质。