Effect of human milk oligosaccharides intervention on gut development and behaviour in piglets

Breastmilk is the optimal and complete source of nutrition for infants during the first 4-6 months of life. Infant formula is the recommended alternative and the first choice for providing nutrition to infants who cannot be exclusively breastfed. However most infant formula is cow’s milk based missing critical nutrients of human milk oligosaccharides (HMOs) . HMOs as prebiotics play a significant role in the development and function of the infant gut microbiota, which in turn affects various aspects of gut development and behavior.

The challenge

Breast-feeding has been linked to higher intelligence in later childhood compared with formula-feeding. Low birth weight infants benefit more in cognitive development from breastfeeding than full term infants. Morphometric brain imaging studies have revealed an increase in the volume of white and sub-cortical grey matter, and parietal lobe cortical thickness, which are associated with IQ in adolescents who were breastfed as infants compared to those who were exclusively formula-fed.

Currently, only ~35.4% of Australian infants are exclusively breastfed until ca. 6 mo. This means that >60% of Australian infants rely on infant formula during early neural development. Early-life nutrition refers to the critical period of the first 1000 days of life, which starts from conception and extends through pregnancy and the first two years of a child's life. This period is widely recognized as a window of opportunity for optimal growth, development, and a child's health and well-being throughout their life. Therefore, our study has the potential to impact neonatal care for all children, particularly those born premature, and reduce costs of health care and social services and will enhance the wealth of Australian societies.

Human milk oligosaccharides (HMOs) are the third most abundant class of biomolecules in mature human milk, after lactose and lipids, reaching levels ~20–25g/L in colostrum and 5–20g/L in mature milk (1, 2). More than 200 different HMOs have been identified (1), with ~10–20% containing the acidic sugar, sialic acid, so called sialylated HMOs. The remaining are neutral HMOs. Human milk contains ~100-1000 fold higher levels of HMOs than bovine milk or any infant formula. Breastmilk is the optimal and complete source of nutrition for infants during the first 4-6 months of life. Infant formula is the recommended alternative and the first choice for providing nutrition to infants who cannot be exclusively breastfed. However most infant formula is cow’s milk based missing critical nutrients HMOs. HMOs as prebiotics play a significant role in the development and function of the infant gut microbiota, which in turn affects various aspects of gut development and behavior. The rate of initial brain growth exceeds that of any other organ or body tissue, and by 2 y of age, the brain reaches 80% of its adult weight. Our recent in vivo magnetic resonance spectroscopic (MRS) studies demonstrated that the sialylated HMOs, 3’sialyllactose (3'-SL) and (6'-SL) can alter the level of important brain metabolites and neurotransmitters required for optimal neurodevelopment in piglets. The most abundant nHMO, 2’-fucosyllactose (2'-FL), improved learning and memory in rodent models. However, we do not know if an acidic, neutral or a combination of the two HMOs can act synergistically to provide an enhanced therapeutical role for neurodevelopment, neuroprotection and gut development of newborn.

Project name
Effect of human milk oligosaccharides (HMOs) intervention on gut development and behaviour in piglets (2023-2025)

Funding Junlebao Dairy Group Co., LTD, stage one project $613,628

Our response

This is the first preclinical study taking a nutraceutical approach to deliver innovative therapeutic applications of key sialylated and neutral or a combination of two human milk oligosaccharides (HMOs) in neurodevelopment, neuroprotection and gut development for newborns. The innovation associated with the assessment of a piglet outcome will allow us to create new knowledge and new insights into the molecular and cellular mechanisms of the therapeutic role of individual HMOs or their combination on brain, gut development, cognitive functions and immunity, with an approach not possible using human subjects.

In 2023 the team successfully completed two piglet’s feeding trials with 32 newborn piglets. More than 12 undergraduate students, 2 PhD students, a research assistant, and staff members participated in this project. All students learned skills including the calculation of daily milk intake, milk preparation, animal feeding, care, animal weighing,  cleaning the animal house, and behavioral testing. The team also successfully completed brain MRI scans of all 32 piglets at the Monash Bioimaging Center.

The goal

The overall goal of our project is to elucidate the molecular and cellular mechanisms of the therapeutic role of dietary sialylated HMOs and neutral HMOs, acting alone or in combination, to enhance neurodevelopment, neuroprotection, gut development and immunity in newborn piglets, an ideal animal model for the human infant.

Our research uses innovative approaches that will provide evidence-based recommendations and new insights into molecular and cellular mechanisms whereby dietary sialyllactose (SL), 2’-fucosyllactose (2’FL) or their combination might optimize neurodevelopment, cognitive function and immunity associated with piglets during early postnatal life.

Our team

Principal scientist

portrait of  Professor Bing Wang
Professor Bing Wang
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Our research team

portrait of Dr Xiaoming Zheng
Dr Xiaoming Zheng
Senior lecturer in in Medical Physics
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portrait of Dr Shaoyu (Shaun) Wang
Dr Shaoyu (Shaun) Wang
Lecturer in in Anatomy & Physiology
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portrait of Dr Allan Gunn
Dr Allan Gunn
A/Professor in Veterinary Reproduction
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Professor Peter Wynn
Adjunct Professor
Dr Wendy Li
Postdoctoral fellow
Mahmudul Amin
PhD student
Dilki Adikari Arachchige
PhD student

Key research publications

  • Jiang, B., Xia, Y., Zhou, L., Liang, X., Chen, X., Chen, M., Li, X., Lin, S., Zhang, N., Zheng, L., Tao, M., Petocz, P., Gallier, S., Rowan, A., & Wang, B. (2022). Safety and tolerance assessment of milk fat globule membrane-enriched infant formulas in healthy term Chinese infants: a randomised multicenter controlled trial. BMC Pediatrics, 22(1), 1-18. [465]. https://doi.org/10.1186/s12887-022-03507-8
  • Madalyn Hobbs, Marefa Jahan, Seyed Ghorashi, Bing Wang. (2021). Current Perspective of Sialylated Milk Oligosaccharides in Mammalian Milk: Implications for Brain and Gut Health of Newborns. Foods 10, 473. https://doi.org/10.3390/foods10020473.
  • Y Xia, PW Jiang, LH Zhou, XH Chen, MZ Chen, XX Li, HB Liu, LY Yang, L Zheng, P Petocz, B Wang, (2021). Neurodevelopmental Outcomes of Healthy Chinese Term Infants Fed Infant Formula Enriched in Bovine Milk Fat Globule Membrane for 12 months: A Randomized Controlled Trial. Asia Pac J Clin Nutr 30(3):401-414 DOI: 10.6133/apjcn.202109_30(3).0007
  • Wang HX, Chen Y, Haque Z, de Veer M, Egan G and Wang B.(2019). Sialylated Milk Oligosaccharides Alter Neurotransmitters and Brain Metabolites Optimising Neurodevelopment in Piglets: An In vivo Magnetic Resonance Spectroscopic (MRS) study. Nutri Neurosci. 20:1-11. DOI: 10.1080/1028415X.2019.1691856
  • CW Yang, PW Zhang, F Wang, B Wang. (2019). Molecular Mechanisms Underlying How Sialyllactose Intervention Promotes Intestinal Maturity by Upregulating GDNF Through a CREB-Dependent Pathway in Neonatal Piglets. Mol Neurobiol, 56:7994-8007. DOI: 10.1007/s12035-019-1628-9
  • Wei JH, Wang ZA, Wang B, Jahan M, Wang ZF, Troy FA II, Du YG. (2018). Characterization of porcine milk oligosaccharides over lactation between primiparous and multiparous female pigs. Scientific Report, 8:4688. doi: 10.1038/s41598-018-23025-x.
  • Gan HA, Zhang QZ, Chen Y, Zhang H, Lin JZ, Kang TS, Zhang JX, Wang B. (2014). Development of New Population-averaged Standard Templates for Spatial Normalization and Segmentation of MR Images for Postnatal Piglet Brains. Magn Reson Imaging. 2014;32:1396-402 doi: 10.1016/j.mri.2014.08.036
  • Wang B. (2012). Molecular Mechanism Underlying Sialic Acid as an Essential Nutrient for Brain Development and Cognition. Adv Nutr. 3:4655-4725.  DOI: 10.3945/an.112.001875
  • Wang B. (2009). Sialic Acid is an essential nutrient for brain development and cognition. Ann Rev Nutr, 29:177-222. DOI: 10.1146/annurev.nutr.28.061807.155515
  • Wang B, Yu B, Karim M, Hu H, Sun Y, Petocz P, Held S, McGreevy P, Brand-Miller J. (2007). Dietary sialic acid supplementation improves learning and memory in piglets. Ame J Clin Nutr 85:561-9. PMID: 17284758. DOI: 10.1093/ajcn/85.2.561
  1. Wang B. Dairy product and process. 30 July 2017, PTC/IB2017/054610
  2. Wang B, Dong ZZ, Composition comprising sialyllactose for use in enhancing learning skills and memory function. Filling date 18 March 2015 PCT/CN15/074457
  3. Wang B, Zheng ZQ, Dong ZZ, Wang JK Lactoferrin and memory and learning speed in Children. Aug 2013 Ref. 12027/EP/EPA P44486/EP, WO2015/028173A1
  4. Wang B Lactoferrin supplementation and diarrhea. Mar 2012 PCT/CN12/072594
  5. Wang B and Faure M. Lactoferrin and white matter Nov 2011 Ref No. EP20110189879/ grant
  6. Wang B, L Fay, J Schmitt et al. Nutrition composition for supporting brain development and function of toddles. June 2010 Ref No. EP2010057661/US13/375,079
  7. Wang B, Faure M, Schmitt J. Lactoferrin and brain health and development in infants. 2009 Ref No. EP2010056237/ US 8524658. PCT/EP2010/056237
  8. Faure M, Wang B, Schmitt J. Lactoferrin and neuronal health and development in infant gut. 2009 Ref No. EP2010056234/ US 13/319,655.
  9. Wang B, Faure M, Schmitt J. Lactoferrin and brain health and protection in adults. 2009 Ref No. PCT/EP2010/056241
  10. Faure M, Wang B, Schmitt J. Lactoferrin and gut neuronal health in adult and/or elderly. 2009 Ref No. PCT/EP2010/056238/ US14/073,522
  11. Wang B, J Brand-Miller. Methods of improving learning and memory in mammals. 2005 Ref No. 19400/09302
  12. Wang B, J Brand-Miller. Method of increasing the salivary sialic acid content in a mammal. 2005 Ref No. 19400/09187/US 11/118,845

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