Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 6 2025 12 13 893 900 EN Sima Taheri Department of Food Science and Technology, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran Morteza Khomeiri Department of Food Science and Technology, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran Hesamaddin Aski Infectious Disease Research Center, Golestan University of Medical Sciences, Gorgan, Iran Hamid Ghoddusi Microbiology Research Unit (MRU), School of Human Sciences, Faculty of Life Sciences and Computing, London Metropolitan University, London, UK Seid Mahdi Jafari Department of Food Science and Technology, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran Ezzat Allah Ghaemi Infectious Disease Research Center, Golestan University of Medical Sciences, Gorgan, Iran 2025 05 20 2025 11 04 Background and Objectives: Human milk provides nutrients, prebiotics, and probiotics that support infants’ physical and mental growth. Human milk microbiota, as a potential source of probiotics and an indicator of the safety of donor milk, is of great importance. Akkermansia muciniphila, a core member of next-generation probiotics (NGPs), with the ability to degrade human milk oligosaccharides (HMOs), may be present in human milk. This study was carried out to assess the total bacterial count and presence of A. muciniphila in raw freshly expressed mothers’ milk and pasteurized donor milk from human milk banks (HMBs). Materials and Methods: 15 raw and 20 pasteurized milk samples were collected and analyzed using a real-time PCR technique with specific primers for A. muciniphila and universal 16S rRNA bacterial primers. Results: Results showed that the average total bacterial count was 4.95 log CFU/ml, which is lower than the normal range reported for human milk. Samples with bacterial count over the standard range were related to the HMBs. Prevalence of A. muciniphila in human milk was 35% and was higher in raw milk samples than in pasteurized samples. Conclusion: In conclusion, raw human milk can serve as a potential source for A. muciniphila isolation as a candidate for NGPs. https://ijm.tums.ac.ir/index.php/ijm/article/view/5502 https://ijm.tums.ac.ir/index.php/ijm/article/download/5502/1836