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Physicochemical Biomolecular Insights into Buffalo Milk-Derived Nanovesicles

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Abstract

Milk is a natural nutraceutical produced by mammals. The nanovesicles of milk play a role in horizontal gene transfer and confer health-benefits to milk consumers. These nanovesicles contain miRNA, mRNA, and proteins which mediate the intercellular communication. In this work, we isolated and characterized the buffalo milk-derived nanovesicles by dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), scanning electron microscopy (SEM), Western probing, and Fourier transform infrared (FTIR) spectroscopy. The DLS data suggested a bimodal size distribution with one mode near 50 nm and the other around 200 nm for the nanovesicles. The NTA and SEM data also supported the size of nanovesicles within a range of 50–200 nm. The FTIR measurements of nanovesicles identified some prominent absorption bands attributable to the proteins (1300–1700 cm−1, amide A and amide B bands), lipids (2800–3100 cm−1), polysaccharides, and nucleic acids (900–1200 cm−1). The comparative expression profiles of immune miRNA signatures (miR-15b, miR-21, miR-27b, miR-125b, miR-155, and miR-500) in nanovesicles isolated from milk, serum, and urine revealed that these miRNAs are present abundantly (P < 0.05) in milk-derived nanovesicles. Milk miRNAs (miR-21 and 500) that were also found stable under different household storage conditions indicated that these could be biologically available to milk consumers. Overall, nanovesicles are a new class of bioactive compounds from buffalo milk with high proportion of stable immune miRNAs compared to urine and plasma of same animals.

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Acknowledgments

The authors are very grateful to Director NDRI, Karnal, for providing the necessary facilities for this study. The authors thank Dr. S. K. Tomar for providing the SEM facility to BV and Dr. Rajan Sharma for allowing VN to use FTIR spectroscopy, and Mark Ware, Kartick Padmanabhan, and Namrata Jain of Malvern Instruments for facilitating the NTA at NDRI.

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    Authors and Affiliations

    1. Molecular Endocrinology, Functional Genomics and Systems Biology Laboratory, Animal Biochemistry Division, ICAR-National Dairy Research Institute (Deemed University), Karnal, 132001, Haryana, India

      Vijay Simha Baddela, Varij Nayan, Payal Rani, Suneel Kumar Onteru & Dheer Singh

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    1. Vijay Simha Baddela
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    2. Varij Nayan
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    3. Payal Rani
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    4. Suneel Kumar Onteru
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    Correspondence to Dheer Singh.

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    Funding

    This work was financially supported by the National Agricultural Science Fund (NASF), formerly known as National Fund for Basic, Strategic, and Frontier Application Research in Agriculture (NFBSFARA) (NFBSFARA/BSA-4006/2013-14) and CRP – Nanotechnology of ICAR, India.

    Conflict of Interest

    The authors declare that they have no conflict of interest.

    Ethical Approval

    Authors have taken approval of institute ethical committee for the collection of blood, urine, and milk from the buffaloes of NDRI cattle and buffalo herd for the sake of research work.

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    Vijay Simha Baddela and Varij Nayan are co-first authors and contributed equally to this work.

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    Baddela, V.S., Nayan, V., Rani, P. et al. Physicochemical Biomolecular Insights into Buffalo Milk-Derived Nanovesicles. Appl Biochem Biotechnol 178, 544–557 (2016). https://doi.org/10.1007/s12010-015-1893-7

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