Human milk as a source of next-generation probiotics: quantifying Akkermansia muciniphila and microbial contamination risks in donor milk
-
Sima Taheri
,
Morteza Khomeiri
,
Hesamaddin Shirzad Aski
,
Hamid Bahador Ghoddusi
,
Seid Mahdi Jafari
,
Ezzat Allah Ghaemi
Abstract
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.
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19. Lackey KA, Williams JE, Meehan CL, Zachek JA, Benda ED, Price WJ, et al. What’s normal? Microbiomes in human milk and infant feces are related to each other but vary geographically: The INSPIRE Study. Front Nutr 2019; 6: 45.
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https://link.springer.com/book/10.1007/978-0-387-68572-4
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22. Selma-Royo M, Calvo Lerma J, Cortés-Macías E, Collado MC. Human milk microbiome: From actual knowledge to future perspective. Semin Perinatol 2021; 45: 151450.
23. Lee S, Heo S, Park M-K, Sung M-H, Jeong D-W. Bacterial community of breast milk in breastfeeding women using culture-dependent and culture-independent approaches. J Microbiol Biotechnol 2024; 34: 2005-2011.
24. Gad S, Sheta MM, Al-Khalafawi AI, Abu El-Fadl HA, Anany M, Sahmoud S, et al. Expressed breast milk contamination in neonatal intensive care unit. Pediatric Health Med Ther 2021; 12: 307-313.
25. Ramsay DT, Kent JC, Owens RA, Hartmann PE. Ultrasound imaging of milk ejection in the breast of lactating women. Pediatrics 2004; 113: 361-367.
26. Landers S, Updegrove K. Bacteriological screening of donor human milk before and after Holder pasteurization. Breastfeed Med 2010; 5: 117-121.
27. Perrin MT, Fogleman AD, Davis DD, Wimer CH, Vogel KG, Palmquist AEL. A pilot study on nutrients, antimicrobial proteins, and bacteria in commerce-free models for exchanging expressed human milk in the USA. Matern Child Nutr 2018; 14 Suppl 6(Suppl 6): e12566.
28. Haiden N, Pimpel B, Assadian O, Binder C, Kreissl A, Repa A, et al. Comparison of bacterial counts in expressed breast milk following standard or strict infection control regimens in neonatal intensive care units: Compliance of mothers does matter. J Hosp Infect 2016; 92: 226-228.
29. Kumar H, du Toit E, Kulkarni A, Aakko J, Linderborg KM, Zhang Y, et al. Distinct patterns in human milk microbiota and fatty acid profiles across specific geographic locations. Front Microbiol 2016; 7: 1619.
30. Cabrera-Rubio R, Collado MC, Laitinen K, Salminen S, Isolauri E, Mira A. The human milk microbiome changes over lactation and is shaped by maternal weight and mode of delivery. Am J Clin Nutr 2012; 96: 544-551.
31. Hermansson H, Kumar H, Collado MC, Salminen S, Isolauri E, Rautava S. Breast milk microbiota is shaped by mode of delivery and intrapartum antibiotic exposure. Front Nutr 2019; 6: 4.
32. Soto A, Martín V, Jiménez E, Mader I, Rodríguez JM, Fernández L. Lactobacilli and bifidobacteria in human breast milk: Influence of antibiotherapy and other host and clinical factors. J Pediatr Gastroenterol Nutr 2014; 59: 78-88.
33. Olivares M, Albrecht S, De Palma G, Ferrer MD, Castillejo G, Schols HA, et al. Human milk composition differs in healthy mothers and mothers with celiac disease. Eur J Nutr 2015; 54: 119-128.
34. de Segura AG, Escuder D, Montilla A, Bustos G, Pallás C, Fernández L, et al. Heating-induced bacteriological and biochemical modifications in human donor milk after Holder pasteurization. J Pediatr Gastroenterol Nutr 2012; 54: 197-203.
35. Miura K, Tanaka M, Date M, Ito M, Mizuno N, Mizuno K. Comparison of bacterial profiles in human milk from mothers of term and preterm infants. Int Breastfeed J 2023; 18: 29.
36. Moossavi S, Sepehri S, Robertson B, Bode L, Goruk S, Field CJ, et al. Composition and variation of the human milk microbiota are influenced by maternal and early-life factors. Cell Host Microbe 2019; 25: 324-335.e4.
37. Aakko J, Kumar H, Rautava S, Wise A, Autran C, Bode L, et al. Human milk oligosaccharide categories define the microbiota composition in human colostrum. Benef Microbes 2017; 8: 563-567.
38. Liao J, Liu Y, Ku T. Changes in physicochemical properties and DNA quality of milk as affected by different heat treatments. Int J Dairy Technol 2018; 71: 333-339.
39. Qi Y, Yu L, Tian F, Zhao J, Zhang H, Chen W, et al. Akkermansia muciniphila supplementation in mice during pregnancy and lactation affects the maternal intestinal microenvironment. Nutrients 2022; 14: 390.
2. Palou A, Picó C. Leptin intake during lactation prevents obesity and affects food intake and food preferences in later life. Appetite 2009; 52: 249-252.
3. Savilahti E, Saarinen KM. Early infant feeding and type 1 diabetes. Eur J Nutr 2009; 48: 243-249.
4. Jeurink PV, van Bergenhenegouwen J, Jiménez E, Knippels LM, Fernández L, Garssen J, et al. Human milk: A source of more life than we imagine. Benef Microbes 2013; 4: 17-30.
5. Gómez-Gallego C, García-Mantrana I, Salminen S, Collado MC. The human milk microbiome and factors influencing its composition and activity. Semin Fetal Neonatal Med 2016; 21: 400-405.
6. Martín R, Langa S, Reviriego C, Jimínez E, Marín ML, Xaus J, et al. Human milk is a source of lactic acid bacteria for the infant gut. J Pediatr 2003; 143: 754-758.
7. Martín R, Jiménez E, Olivares M, Marín ML, Fernández L, Xaus J, et al. Lactobacillus salivarius CECT 5713, a potential probiotic strain isolated from infant feces and breast milk of a mother-child pair. Int J Food Microbiol 2006; 112: 35-43.
8. Martín R, Jiménez E, Heilig H, Fernández L, Marín ML, Zoetendal EG, et al. Isolation of bifidobacteria from breast milk and assessment of the bifidobacterial population by PCR-denaturing gradient gel electrophoresis and quantitative real-time PCR. Appl Environ Microbiol 2009; 75: 965-969.
9. Marcobal A, Barboza M, Froehlich JW, Block DE, German JB, Lebrilla CB, et al. Consumption of human milk oligosaccharides by gut-related microbes. J Agric Food Chem 2010; 58: 5334-5340.
10. Shirvanian M, Azimian Zavareh V, Zamanzadeh Z, Janghorban M, Mohammadi E, Ahangarzadeh S. Probiotic potential, antiproliferative, and proapoptotic activities of Lactobacillus fermentum isolated from breast milk. Adv Biomed Res 2023; 12: 242.
11. Malekzadeh J, Synaii S, Ebrahimzadeh Koor B, Falsafian G, Nakhaie MR. Growth indices of exclusively breastfed until 6 months age and formula-fed infants in southwest of Iran. Int J Prev Med 2019; 10: 207.
12. Moro GE, Billeaud C, Rachel B, Calvo J, Cavallarin L, Christen L, et al. Processing of donor human milk: Update and recommendations from the European Milk Bank Association (EMBA). Front Pediatr 2019; 7: 49.
13. Hamidi N, Mousavi SS, Farahani LA, Khosravi A, Saboute M. Comparison of the short-term neonatal outcomes of preterm neonates before and after the launch of human milk bank in Iran: A retrospective descriptive study. BMC Pediatr 2025; 25: 202.
14. Fernández L, Langa S, Martín V, Maldonado A, Jiménez E, Martín R, et al. The human milk microbiota: Origin and potential roles in health and disease. Pharmacol Res 2013; 69: 1-10.
15. National Institute for Health and Clinical Excellence. Donor Breast Milk Banks: The Operation of Donor Milk Bank Services. London: NICE; 2010.
16. Conboy-Stephenson R, Ross RP, Kelly AL, Stanton C. Donor human milk: The influence of processing technologies on its nutritional and microbial composition. Front Nutr 2024; 11: 1468886.
17. Derrien M, Vaughan EE, Plugge CM, de Vos WM. Akkermansia muciniphila gen. nov., sp. nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol 2004; 54: 1469-1476.
18. Urbaniak C, Cummins J, Brackstone M, Macklaim JM, Gloor GB, Baban CK, et al. Microbiota of human breast tissue. Appl Environ Microbiol 2014; 80: 3007-3014.
19. Lackey KA, Williams JE, Meehan CL, Zachek JA, Benda ED, Price WJ, et al. What’s normal? Microbiomes in human milk and infant feces are related to each other but vary geographically: The INSPIRE Study. Front Nutr 2019; 6: 45.
20. Krieg NR, Staley JT, Brown DR, Hedlund B, Paster B, Ward N, et al (2010). Bergey's Manual of Systematic Bacteriology, Volume 4: The Bacteroidetes, Spirochaetes, Tenericutes (Mollicutes), Acidobacteria, Fibrobacteres, Fusobacteria, Dictyoglomi, Gemmatimonadetes, Lentisphaerae, Verrucomicrobia, Chlamydiae, and Planctomycetes.
https://link.springer.com/book/10.1007/978-0-387-68572-4
21. Collado MC, Derrien M, Isolauri E, de Vos WM, Salminen S. Intestinal integrity and Akkermansia muciniphila, a mucin-degrading member of the intestinal microbiota present in infants, adults, and the elderly. Appl Environ Microbiol 2007; 73: 7767-7770.
22. Selma-Royo M, Calvo Lerma J, Cortés-Macías E, Collado MC. Human milk microbiome: From actual knowledge to future perspective. Semin Perinatol 2021; 45: 151450.
23. Lee S, Heo S, Park M-K, Sung M-H, Jeong D-W. Bacterial community of breast milk in breastfeeding women using culture-dependent and culture-independent approaches. J Microbiol Biotechnol 2024; 34: 2005-2011.
24. Gad S, Sheta MM, Al-Khalafawi AI, Abu El-Fadl HA, Anany M, Sahmoud S, et al. Expressed breast milk contamination in neonatal intensive care unit. Pediatric Health Med Ther 2021; 12: 307-313.
25. Ramsay DT, Kent JC, Owens RA, Hartmann PE. Ultrasound imaging of milk ejection in the breast of lactating women. Pediatrics 2004; 113: 361-367.
26. Landers S, Updegrove K. Bacteriological screening of donor human milk before and after Holder pasteurization. Breastfeed Med 2010; 5: 117-121.
27. Perrin MT, Fogleman AD, Davis DD, Wimer CH, Vogel KG, Palmquist AEL. A pilot study on nutrients, antimicrobial proteins, and bacteria in commerce-free models for exchanging expressed human milk in the USA. Matern Child Nutr 2018; 14 Suppl 6(Suppl 6): e12566.
28. Haiden N, Pimpel B, Assadian O, Binder C, Kreissl A, Repa A, et al. Comparison of bacterial counts in expressed breast milk following standard or strict infection control regimens in neonatal intensive care units: Compliance of mothers does matter. J Hosp Infect 2016; 92: 226-228.
29. Kumar H, du Toit E, Kulkarni A, Aakko J, Linderborg KM, Zhang Y, et al. Distinct patterns in human milk microbiota and fatty acid profiles across specific geographic locations. Front Microbiol 2016; 7: 1619.
30. Cabrera-Rubio R, Collado MC, Laitinen K, Salminen S, Isolauri E, Mira A. The human milk microbiome changes over lactation and is shaped by maternal weight and mode of delivery. Am J Clin Nutr 2012; 96: 544-551.
31. Hermansson H, Kumar H, Collado MC, Salminen S, Isolauri E, Rautava S. Breast milk microbiota is shaped by mode of delivery and intrapartum antibiotic exposure. Front Nutr 2019; 6: 4.
32. Soto A, Martín V, Jiménez E, Mader I, Rodríguez JM, Fernández L. Lactobacilli and bifidobacteria in human breast milk: Influence of antibiotherapy and other host and clinical factors. J Pediatr Gastroenterol Nutr 2014; 59: 78-88.
33. Olivares M, Albrecht S, De Palma G, Ferrer MD, Castillejo G, Schols HA, et al. Human milk composition differs in healthy mothers and mothers with celiac disease. Eur J Nutr 2015; 54: 119-128.
34. de Segura AG, Escuder D, Montilla A, Bustos G, Pallás C, Fernández L, et al. Heating-induced bacteriological and biochemical modifications in human donor milk after Holder pasteurization. J Pediatr Gastroenterol Nutr 2012; 54: 197-203.
35. Miura K, Tanaka M, Date M, Ito M, Mizuno N, Mizuno K. Comparison of bacterial profiles in human milk from mothers of term and preterm infants. Int Breastfeed J 2023; 18: 29.
36. Moossavi S, Sepehri S, Robertson B, Bode L, Goruk S, Field CJ, et al. Composition and variation of the human milk microbiota are influenced by maternal and early-life factors. Cell Host Microbe 2019; 25: 324-335.e4.
37. Aakko J, Kumar H, Rautava S, Wise A, Autran C, Bode L, et al. Human milk oligosaccharide categories define the microbiota composition in human colostrum. Benef Microbes 2017; 8: 563-567.
38. Liao J, Liu Y, Ku T. Changes in physicochemical properties and DNA quality of milk as affected by different heat treatments. Int J Dairy Technol 2018; 71: 333-339.
39. Qi Y, Yu L, Tian F, Zhao J, Zhang H, Chen W, et al. Akkermansia muciniphila supplementation in mice during pregnancy and lactation affects the maternal intestinal microenvironment. Nutrients 2022; 14: 390.
| Files | ||
| Issue | Vol 17 No 6 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i6.20356 | |
| Keywords | ||
| Akkermansia muciniphila Bacterial counts Human milk Microbiota Pasteurization Real-time polymerase chain reaction | ||
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How to Cite
1.
Taheri S, Khomeiri M, Aski H, Ghoddusi H, Jafari SM, Ghaemi EA. Human milk as a source of next-generation probiotics: quantifying Akkermansia muciniphila and microbial contamination risks in donor milk. Iran J Microbiol. 2025;17(6):893-900.
