Brunelli Laura, Perotti Susanna, Gargari Giorgio, De Vitis Valerio, Mantegazza Giacomo, Ferrari Roberto, Minuzzo Mario, Pierallini Elena, Ricci Giovanni, Fiore Walter, Guglielmetti Simone
Alfasigma S.p.A., Milan, Italy.
Department of Food, Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy.
Appl Environ Microbiol. 2025 May 21;91(5):e0239424. doi: 10.1128/aem.02394-24. Epub 2025 Apr 24.
The commercialization of a probiotic microorganism requires genetic and phenotypic consistency across production lots. However, the large-scale industrial production of probiotic microbial biomasses over years poses a risk of genetic drift, potentially affecting the probiotic's ability to confer health benefits. This study assessed the stability of DG (DSM 34154), a commercial probiotic. Seven isolates from different commercial lots over 10 years, one 8-year-old laboratory subculture, one isolate from human feces, and the DG strain deposited at DSMZ were analyzed. Shotgun and PacBio sequencing were combined to obtain the complete genome of the 10 isolates. Comparative analysis showed that the seven commercial isolates had identical genomes, differing from the DSMZ isolate by one synonymous transition and one non-synonymous transversion. The laboratory subculture strain had two additional mutations. Phenotypic analyses, including antibiotic resistance, carbohydrate fermentation profile, survival to simulated gastrointestinal transit, immunomodulatory capacity, and radical scavenging ability, found no significant differences among isolates. Overall, this study demonstrates the substantial equivalence of DG over 10 years of industrial production, indicating that the current industrial practices help prevent genome alterations that could compromise probiotic performance. Similar studies should be part of continuous monitoring and quality control measures for the probiotic products on the market.IMPORTANCEThe genetic and functional stability of probiotic strains during years of industrial production is essential but has not been clearly demonstrated for many strains. This study shows that careful industrial practices can maintain the genetic integrity and functionality of probiotics. Using advanced genome sequencing and detailed laboratory tests, we confirmed that the probiotic DG (DSM 34154) has remained stable over a decade of production, consistently delivering its health-promoting properties. These findings support the quality and reliability of probiotic products, fostering consumer trust and highlighting the importance of continuous monitoring in probiotic manufacturing to sustain quality assurance.
益生菌微生物的商业化需要各生产批次间具备基因和表型的一致性。然而，多年来益生菌微生物生物量的大规模工业化生产存在基因漂移的风险，这可能会影响益生菌产生健康益处的能力。本研究评估了商业益生菌DG（DSM 34154）的稳定性。分析了来自10年间不同商业批次的7株分离株、一株8年的实验室传代培养物、一株来自人类粪便的分离株以及保藏于德国微生物和细胞培养物保藏中心（DSMZ）的DG菌株。结合鸟枪法测序和PacBio测序以获得这10株分离株的完整基因组。比较分析表明，7株商业分离株具有相同的基因组，与DSMZ分离株存在一个同义转换和一个非同义颠换的差异。实验室传代培养菌株还有另外两个突变。包括抗生素抗性、碳水化合物发酵谱、模拟胃肠道转运存活能力、免疫调节能力和自由基清除能力在内的表型分析发现，各分离株之间没有显著差异。总体而言，本研究证明了DG在10年工业生产过程中的实质等效性，表明当前的工业生产实践有助于防止可能损害益生菌性能的基因组改变。类似的研究应成为市场上益生菌产品持续监测和质量控制措施的一部分。重要性益生菌菌株在多年工业生产过程中的遗传和功能稳定性至关重要，但许多菌株尚未得到明确证明。本研究表明，谨慎的工业生产实践可以维持益生菌的遗传完整性和功能。通过使用先进的基因组测序和详细的实验室测试，我们证实益生菌DG（DSM 34154）在十多年的生产过程中一直保持稳定，始终发挥其促进健康的特性。这些发现支持了益生菌产品的质量和可靠性，增强了消费者的信任，并突出了在益生菌生产中持续监测以维持质量保证的重要性。
