Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Advertisement

Scientific Reports
  • View all journals
  • Search
  • My Account Login
  • Content Explore content
  • About the journal
  • Publish with us
  • Sign up for alerts
  • RSS feed
  1. nature
  2. scientific reports
  3. articles
  4. article
Modulation of fat mobilization and adipose tissue gene expression in Holstein cows supplemented with omega-3 fatty acids and N-acetyl-tryptophan during the transition period
Download PDF
Download PDF
  • Article
  • Open access
  • Published: 08 February 2026

Modulation of fat mobilization and adipose tissue gene expression in Holstein cows supplemented with omega-3 fatty acids and N-acetyl-tryptophan during the transition period

  • Mansoureh Ghorbanalinia1,
  • Essa Dirandeh1,
  • Zarbakht Ansari-Pirsaraei1,
  • Hasan Sadri2 &
  • …
  • William W Thatcher3 

Scientific Reports , Article number:  (2026) Cite this article

  • 583 Accesses

  • Metrics details

We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Biochemistry
  • Endocrinology
  • Molecular biology
  • Physiology

Abstract

The transition period is critical for dairy cows, characterized by negative energy balance (NEB), excessive adipose mobilization, and metabolic challenges. This study investigated the effects of dietary omega-3 fatty acids (O3) and N-acetyl-tryptophan (NAT) on blood metabolites and adipose tissue gene expression in Holstein cows. Forty-eight multiparous cows were assigned to four groups (control, O3, NAT, O3+NAT) from -21 to +42 days relative to calving. Body weight (BW) and body condition score (BCS) were recorded, and plasma non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), insulin, and glucose were measured at -21, 0 (calving), +21, and +42 days. Adipose biopsies at +21 and +42 days postpartum were used to perform qPCR analysis of lipogenesis-related genes (Acetyl-CoA carboxylase alpha (ACACA), peroxisome proliferator-activated receptor gamma (PPARγ), lipoprotein lipase (LPL)), fatty acid oxidation (Acyl-CoA oxidase 1 (ACOX1)), lipolysis (hormone-sensitive lipase (LIPE), adipose triglyceride lipase (ATGL)), and adiponectin receptors (AdipoR1, AdipoR2). Statistical analysis used two-way ANOVA with repeated measures. Cows supplemented with O3+NAT maintained higher BW (p<0.05) and BCS (p<0.05), exhibited lower NEFA and BHBA (p<0.05), and had increased insulin (p<0.05) and tended to have higher glucose (p=0.08) compared with controls. PPARγ, LPL, and adiponectin receptors (AdipoR1, AdipoR2) were upregulated in all supplemented groups compared to controls (p<0.05). ACOX1 was downregulated in O3, NAT, and O3+NAT groups compared to control (p<0.05). Network analysis revealed strong positive correlations between insulin and AdipoR1/2 (r>0.7) and positive correlations between NEFA/BHBA and LIPE/β2AR (r>0.75) at d 21. These results demonstrate that O3 and NAT act via complementary mechanisms to attenuate lipolysis, promote lipid storage, and enhance metabolic homeostasis during the transition period.

Data availability

The data that support the findings of this study are available, on reasonable request, from the corresponding author.

References

  1. Drackley, J. K. Biology of dairy cows during the transition period: the final frontier?. J. Dairy Sci. 82, 2259–73. https://doi.org/10.3168/jds.S0022-0302(99)75474-3 (1999).

    Google Scholar 

  2. Ingvartsen, K. L. & Moyes, K. Nutrition, immune function and health of dairy cattle. Animal. 7, 112–22. https://doi.org/10.1017/S175173111200170X (2013).

    Google Scholar 

  3. Overton, T. R., McArt, J. A. A. & Nydam, D. V. A 100-Year Review: Metabolic health indicators and management of dairy cattle. J. Dairy Sci. 100, 10398–417. https://doi.org/10.3168/jds.2017-13054 (2017).

    Google Scholar 

  4. Sordillo, L. M. & Mavangira, V. The nexus between nutrient metabolism, oxidative stress and inflammation in transition cows. Anim. Prod. Sci. 54, 1204–14. https://doi.org/10.1071/AN14503 (2014).

    Google Scholar 

  5. Roche, J. R., Bell, A. W., Overton, T. R. & Loor, J. J. Nutritional management of the transition cow in the 21st century—a paradigm shift in thinking. Anim. Prod. Sci. 53, 1000–23. https://doi.org/10.1071/AN12293 (2013).

    Google Scholar 

  6. Esposito, G., Irons, P. C., Webb, E. C. & Chapwanya, A. Interactions between negative energy balance, metabolic diseases, uterine health and immune response in transition dairy cows. Anim. Reprod. Sci. 144, 60–71. https://doi.org/10.1016/j.anireprosci.2013.11.007 (2014).

    Google Scholar 

  7. Zachut, M. et al. Periparturient dairy cows do not exhibit hepatic insulin resistance, yet adipose-specific insulin resistance occurs in cows prone to high weight loss. J. Dairy Sci. 96, 5656–69. https://doi.org/10.3168/jds.2012-6202 (2013).

    Google Scholar 

  8. Dirandeh, E., Ghorbanalinia, M., Roodbari, A. R. & Colazo, M. G. Effects of dietary conjugated linoleic acid on metabolic status, BW and expression of genes related to lipid metabolism in adipose tissue of dairy cows during peripartum. Animal. 15, 100105. https://doi.org/10.1016/j.animal.2020.100105 (2021).

    Google Scholar 

  9. Mohammed, A. A., Al-Saiady, M., El-Waziry, A. & Alshaheen, T. Effects of dietary omega-3 fatty acids on reproductive performance and biochemical parameters of lactating cows in arid subtropics. Pakistan J. Zool. 56, 123–30. https://doi.org/10.17582/journal.pjz/2024.56.1.123.130 (2024).

    Google Scholar 

  10. Coyne, G. S., Kenny, D. A., Childs, S., Sreenan, J. M. & Waters, S. M. Dietary n-3 polyunsaturated fatty acids alter the expression of genes involved in prostaglandin biosynthesis in the bovine uterus. Theriogenology. 70, 772–82. https://doi.org/10.1016/j.theriogenology.2008.05.048 (2008).

    Google Scholar 

  11. Kuda, O., Rossmeisl, M. & Kopecky, J. Omega-3 fatty acids and adipose tissue biology. Mol. Aspects Med. 64, 147–60. https://doi.org/10.1016/j.mam.2018.01.004 (2018).

    Google Scholar 

  12. Mohammed, A. A., Al-Saiady, M., El-Waziry, A. & Alshaheen, T. Effects of dietary omega-3 fatty acids on reproductive performance and biochemical parameters of lactating cows in arid subtropics. Pakistan J. Zool. 56, 123–30. https://doi.org/10.17582/journal.pjz/2024.56.1.123.130 (2024).

    Google Scholar 

  13. Mohammed, A. A., Al-Saiady, M., El-Waziry, A. & Alshaheen, T. Effects of dietary omega-3 fatty acids on reproductive performance and biochemical parameters of lactating cows in arid subtropics. Pakistan J. Zool. 56, 123–30. https://doi.org/10.17582/journal.pjz/2024.56.1.123.130 (2024).

    Google Scholar 

  14. Coyne, G. S., Kenny, D. A., Childs, S., Sreenan, J. M. & Waters, S. M. Dietary n-3 polyunsaturated fatty acids alter the expression of genes involved in prostaglandin biosynthesis in the bovine uterus. Theriogenology. 70, 772–82. https://doi.org/10.1016/j.theriogenology.2008.05.048 (2008).

    Google Scholar 

  15. Kra, G. et al. Alpha-linolenic acid modulates systemic and adipose tissue-specific insulin sensitivity, inflammation, and the endocannabinoid system in dairy cows. Sci. Rep. 13, 5280. https://doi.org/10.1038/s41598-023-32456-7 (2023).

    Google Scholar 

  16. Abou-Rjeileh, U. et al. Oleic acid abomasal infusion limits lipolysis and improves insulin sensitivity in adipose tissue from periparturient dairy cows. J. Dairy Sci. 106, 4306–23. https://doi.org/10.3168/jds.2022-22699 (2023).

    Google Scholar 

  17. Richard, D. M. et al. L-tryptophan: basic metabolic functions, behavioral research and therapeutic indications. Int. J. Tryptophan. Res. 2, 45–60. https://doi.org/10.4137/IJTR.S2129 (2009).

    Google Scholar 

  18. Jo, J. H. et al. Effects of rumen-protected L-Tryptophan supplementation on productivity, physiological indicators, blood profiles, and heat shock protein gene expression in lactating holstein cows under heat stress conditions. Int. J. Mol. Sci. 25, 1217. https://doi.org/10.3390/ijms25021217 (2024).

    Google Scholar 

  19. Yamauchi, T., Iwabu, M., Okada-Iwabu, M. & Kadowaki, T. Adiponectin receptors: a review of their structure, function and how they work. Best Pract. Res. Clin. Endocrinol. Metab. 28, 15–23. https://doi.org/10.1016/j.beem.2013.09.003 (2014).

    Google Scholar 

  20. Liu, X. et al. Effects of Prepartum L-Tryptophan supplementation on the postpartum performance of holstein cows. Animals. 14, 1278. https://doi.org/10.3390/ani14091278 (2024).

    Google Scholar 

  21. Luo, Z. et al. Association between tryptophan metabolism and inflammatory biomarkers in dairy cows with ketosis. Metabolites. 13, 123. https://doi.org/10.3390/metabo13020123 (2023).

    Google Scholar 

  22. Dirandeh, E., Ghorbanalinia, M., Rezaei-Roodbari, A. & Colazo, M. G. Relationship between body condition score loss and mRNA of genes related to fatty acid metabolism and the endocannabinoid system in adipose tissue of periparturient cows. Animal. 14, 1724–32. https://doi.org/10.1017/S1751731120000474 (2020).

    Google Scholar 

  23. Ning, M. et al. Gene co-expression network and differential expression analyses of subcutaneous white adipose tissue reveal novel insights into the pathological mechanisms underlying ketosis in dairy cows. J. Dairy Sci. 106, 5018–28. https://doi.org/10.3168/jds.2022-22954 (2023).

    Google Scholar 

  24. Edmonson, A. J., Lean, I. J., Weaver, L. D., Farver, T. & Webster, G. A body condition scoring chart for Holstein dairy cows. J. Dairy Sci. 72, 68–78. https://doi.org/10.3168/jds.S0022-0302(89)79081-0 (1989).

    Google Scholar 

  25. Zachut, M. et al. Periparturient dairy cows do not exhibit hepatic insulin resistance, yet adipose-specific insulin resistance occurs in cows prone to high weight loss. J. Dairy Sci. 96, 5656–69. https://doi.org/10.3168/jds.2012-6202 (2013).

    Google Scholar 

  26. Bustin, S. A. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin. Chem. 55, 611–22. https://doi.org/10.1373/clinchem.2008.112797 (2009).

    Google Scholar 

  27. Miller, M. L. Effects of Omega-3 fatty acids on milk production responses and milk and plasma fatty acids in dairy cows. J. Dairy Sci. 107, 123–35. https://doi.org/10.3168/jds.2023-24112 (2024).

    Google Scholar 

  28. Choi, W. T., Ghassemi Nejad, J., Moon, J. O. & Lee, H. G. Dietary supplementation of acetate-conjugated tryptophan alters feed intake, milk yield and composition, blood profile, physiological variables, and heat shock protein gene expression in heat-stressed dairy cows. J. Therm. Biol. 98, 102949. https://doi.org/10.1016/j.jtherbio.2021.102949 (2021).

    Google Scholar 

  29. Liu, G. et al. Dietary tryptophan supplementation improves antioxidant status and alleviates inflammation, endoplasmic reticulum stress, apoptosis, and pyroptosis in the intestine of piglets after lipopolysaccharide challenge. Antioxidants (Basel). 11, 123. https://doi.org/10.3390/antiox11010123 (2022).

    Google Scholar 

  30. Tomczyk, M. et al. Omega-3 fatty acid supplementation affects tryptophan metabolism during a 12-week endurance training in amateur runners: a randomized controlled trial. Sci. Rep. 14, 4102. https://doi.org/10.1038/s41598-024-54102-8 (2024).

    Google Scholar 

  31. Yamauchi, T., Iwabu, M., Okada-Iwabu, M. & Kadowaki, T. Adiponectin receptors: a review of their structure, function and how they work. Best Pract. Res. Clin. Endocrinol. Metab. 28, 15–23. https://doi.org/10.1016/j.beem.2013.09.003 (2014).

    Google Scholar 

  32. Mezzetti, M., Piccioli-Cappelli, F., Minuti, A. & Trevisi, E. Effects of an intravenous infusion of emulsified fish oil rich in long-chained omega-3 fatty acids on plasma total fatty acids profile, metabolic conditions, and performances of postpartum dairy cows during the early lactation. Front. Vet. Sci. 9, 870901. https://doi.org/10.3389/fvets.2022.870901 (2022).

    Google Scholar 

  33. Conejos, J. R. V. et al. Supplementing with L-Tryptophan increases medium protein and alters expression of genes and proteins involved in milk protein synthesis and energy metabolism in bovine mammary cells. Int. J. Mol. Sci. 22, 123. https://doi.org/10.3390/ijms22010123 (2021).

    Google Scholar 

  34. Field, S. L., Ouellet, V., Sheftel, C. M., Hernandez, L. L. & Laporta, J. In vitro effects of 5-Hydroxy-L-tryptophan supplementation on primary bovine mammary epithelial cell gene expression under thermoneutral or heat shock conditions. Sci. Rep. 12, 3820. https://doi.org/10.1038/s41598-022-07836-6 (2022).

    Google Scholar 

  35. Jump, D. B. et al. Docosahexaenoic acid (DHA) and hepatic gene transcription. Chem. Phys. Lipids. 153, 3–13. https://doi.org/10.1016/j.chemphyslip.2008.02.001 (2008).

    Google Scholar 

  36. Kra, G. et al. Effects of omega-3 supplementation on components of the endocannabinoid system and metabolic and inflammatory responses in adipose and liver of peripartum dairy cows. J. Anim. Sci. Biotechnol. 13, 114. https://doi.org/10.1186/s40104-022-00749-0 (2022).

    Google Scholar 

  37. Van Veldhoven, P. P. Biochemistry and genetics of inherited disorders of peroxisomal fatty acid metabolism. J Lipid Res. 51, 2863–2895. https://doi.org/10.1194/jlr.R005959 (2010).

    Google Scholar 

  38. Bauman, D. E. & Griinari, J. M. Nutritional regulation of milk fat synthesis. Annu. Rev. Nutr. 23, 203–27. https://doi.org/10.1146/annurev.nutr.23.011702.073408 (2003).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Animal Science, Sari Agricultural Sciences and Natural Resources University, P. O. Box: 578, Sari, Mazandaran, Iran

    Mansoureh Ghorbanalinia, Essa Dirandeh & Zarbakht Ansari-Pirsaraei

  2. Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, 5166616471, Iran

    Hasan Sadri

  3. Department of Animal Science, University of Florida, P. O. Box: 511, Florida, USA

    William W Thatcher

Authors
  1. Mansoureh Ghorbanalinia
    View author publications

    Search author on:PubMed Google Scholar

  2. Essa Dirandeh
    View author publications

    Search author on:PubMed Google Scholar

  3. Zarbakht Ansari-Pirsaraei
    View author publications

    Search author on:PubMed Google Scholar

  4. Hasan Sadri
    View author publications

    Search author on:PubMed Google Scholar

  5. William W Thatcher
    View author publications

    Search author on:PubMed Google Scholar

Contributions

Essa Dirandeh: Writing – review & editing, Supervision, Project administration, Methodology, Data curation, Conceptualization. Mansoureh Ghorbanalinia: Performing project Resources. Zarbakht Ansari-Pirsaraei: Formal analysis. William W. Thatcher and Hasan Sadri: review & editing.

Corresponding author

Correspondence to Essa Dirandeh.

Ethics declarations

Competing interest

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ghorbanalinia, M., Dirandeh, E., Ansari-Pirsaraei, Z. et al. Modulation of fat mobilization and adipose tissue gene expression in Holstein cows supplemented with omega-3 fatty acids and N-acetyl-tryptophan during the transition period. Sci Rep (2026). https://doi.org/10.1038/s41598-026-38923-8

Download citation

  • Received: 14 September 2025

  • Accepted: 01 February 2026

  • Published: 08 February 2026

  • DOI: https://doi.org/10.1038/s41598-026-38923-8

Share this article

Anyone you share the following link with will be able to read this content:

Sorry, a shareable link is not currently available for this article.

Provided by the Springer Nature SharedIt content-sharing initiative

Keywords

  • Dairy cows
  • Negative energy balance
  • Lipolysis
  • Omega-3 fatty acids
  • N-acetyl tryptophan
  • Adipose tissue
  • Gene expression
Download PDF

Advertisement

Explore content

  • Research articles
  • News & Comment
  • Collections
  • Subjects
  • Follow us on Facebook
  • Follow us on X
  • Sign up for alerts
  • RSS feed

About the journal

  • About Scientific Reports
  • Contact
  • Journal policies
  • Guide to referees
  • Calls for Papers
  • Editor's Choice
  • Journal highlights
  • Open Access Fees and Funding

Publish with us

  • For authors
  • Language editing services
  • Open access funding
  • Submit manuscript

Search

Advanced search

Quick links

  • Explore articles by subject
  • Find a job
  • Guide to authors
  • Editorial policies

Scientific Reports (Sci Rep)

ISSN 2045-2322 (online)

nature.com sitemap

About Nature Portfolio

  • About us
  • Press releases
  • Press office
  • Contact us

Discover content

  • Journals A-Z
  • Articles by subject
  • protocols.io
  • Nature Index

Publishing policies

  • Nature portfolio policies
  • Open access

Author & Researcher services

  • Reprints & permissions
  • Research data
  • Language editing
  • Scientific editing
  • Nature Masterclasses
  • Research Solutions

Libraries & institutions

  • Librarian service & tools
  • Librarian portal
  • Open research
  • Recommend to library

Advertising & partnerships

  • Advertising
  • Partnerships & Services
  • Media kits
  • Branded content

Professional development

  • Nature Awards
  • Nature Careers
  • Nature Conferences

Regional websites

  • Nature Africa
  • Nature China
  • Nature India
  • Nature Japan
  • Nature Middle East
  • Privacy Policy
  • Use of cookies
  • Legal notice
  • Accessibility statement
  • Terms & Conditions
  • Your US state privacy rights
Springer Nature

© 2026 Springer Nature Limited

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing