ArticleFrontiers in microbiology2024
An optimized culturomics strategy for isolation of human milk microbiota.
Article in Frontiers in microbiology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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9 citing papers in PubMed, 16 citations in OpenAlex.
- Traditional Fermented Dairy Products as Reservoirs of Bifidobacterium With Probiotic Potential: From Microbial Diversity to Functional Characterization.Comprehensive reviews in food science and food safety · 2026Review
- Infant gut microbiota coverage by different culture media.BMC microbiology · 2026Article
- Unveiling functional heterogeneity in pasteurizedFrontiers in nutrition · 2026Article
- New insights into the cultivability of human milk bacteria from ingestion to digestion and implications for their Immunomodulatory properties.Scientific reports · 2025Article
- Integrating the milk microbiome signatures in mastitis: milk-omics and functional implications.World journal of microbiology & biotechnology · 2025Review
- Discovery and cultivation of prokaryotic taxa in the age of metagenomics and artificial intelligence.The ISME journal · 2025Article
- Human breast milk and its impact on inflammatory bowel disease.Journal of smooth muscle research = Nihon Heikatsukin Gakkai kikanshi · 2025Review
- Bacterial Community of Breast Milk in Breastfeeding Women Using Culture-Dependent and Culture-Independent Approaches.Journal of microbiology and biotechnology · 2024Article
- Identification and Characterization of Human Breast Milk and Infant Fecal Cultivable Lactobacilli Isolated in Bulgaria: A Pilot Study.Microorganisms · 2024Article
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Authors and funding
9 authors.
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Abstract
Viable microorganisms and a diverse microbial ecosystem found in human milk play a crucial role in promoting healthy immune system and shaping the microbial community in the infant's gut. Culturomics is a method to obtain a comprehensive repertoire of human milk microbiota. However, culturomics is an onerous procedure, and needs expertise, making it difficult to be widely implemented. Currently, there is no efficient and feasible culturomics method specifically designed for human milk microbiota yet. Therefore, the aim of this study was to develop a more efficient and feasible culturomics method specifically designed for human milk microbiota. We obtained fresh samples of human milk from healthy Chinese mothers and conducted a 27-day enrichment process using blood culture bottles. Bacterial isolates were harvested at different time intervals and cultured on four different types of media. Using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, we identified a total of 6601 colonies and successfully obtained 865 strains, representing 4 phyla, 21 genera, and 54 species. By combining CBA and MRS media, we were able to cultivate over 94.4% of bacterial species with high diversity, including species-specific microorganisms. Prolonged pre-incubation in blood culture bottles significantly increased the number of bacterial species by about 33% and improved the isolation efficiency of beneficial bacteria with low abundance in human milk. After optimization, we reduced the pre-incubation time in blood culture bottles and selected optimal picking time-points (0, 3, and 6 days) at 37°C. By testing 6601 colonies using MALDI-TOF MS, we estimated that this new protocol could obtain more than 90% of bacterial species, reducing the workload by 57.0%. In conclusion, our new culturomics strategy, which involves the combination of CBA and MRS media, extended pre-incubation enrichment, and optimized picking time-points, is a feasible method for studying the human milk microbiota. This protocol significantly improves the efficiency of culturomics and allows for the establishment of a comprehensive repertoire of bacterial species and strains in human milk.
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