MILK BIOMARKERS AS INDICATORS OF METABOLIC STATUS IN DAIRY COWS

4th International Symposium On Biotechnology (2026),  [pp. 365-381]
 
AUTHOR(S) / АУТОР(И): Radojica Đoković , Biljana Anđelić , Marko Cincović , Milun D. Petrović , Miloš Ži. Petrović , Boban Jašović , Aleksandar Čukić 
 
Download Full Pdf   

DOI: https://doi.org/10.46793/SBT26.365D

ABSTRACT / САЖЕТАК:

This study aimed to determine milk biomarkers in dairy cows for early diagnosis of subclinical metabolic disease at different stages of lactation  on the basis relationships between milk and blood metabolites. Blood and milk samples were collected from 100 Holstein dairy cows during morning. The cows were allocated to four groups according to the production period, including Group 1 – cows in early lactation (n=18), Group 2 – cows in full lactation (n=26), Group 3 – cows in mid lactation (n=25), and Group 4 – cows in late lactation (n=31). Chemical composition of cows milk was analysed as well as biochemical parameters in serum. Statistically significant differences (p<0.01) were found between the experimental groups of cows in blood NEFA, BHB, TP, TG, TChol, urea, globulin, AST, ALT, ALP, GGT and LDH levels, milk fat, lactose and protein concentrations, and enzyme activities. Milk enzymes were positively correlated (p<0.05) with blood enzyme activities and blood NEFA, BHB and TBil concentrations, and negatively correlated (p<0.05) with blood glucose, TChol and TG levels. Blood and milk BHB values were markedly positively correlated (p<0.01). Milk fat was positively correlated (p<0.05) with glucose, TP and TG levels, and negatively correlated (p < 0.05) with BHB, NEFA, TBil, ALT, LDH and ALP in the blood. The results showed that milk fat, BHB and milk enzyme activity in the milk can be used to monitor metabolic status in dairy cows at the herd level, and can serve as biomarkers for early detection of subclinical metabolic disease.

KEYWORDS / КЉУЧНЕ РЕЧИ:

cows, lactation, blood, milk, biomarkers, correlations

ACKNOWLEDGEMENT / ПРОЈЕКАТ:

The research presented in this article is part of No. 451-03-33/2026-03/ 200088 financially supported by Ministry of Science, Technological Development and Innovation of Republic of Serbia.

REFERENCES / ЛИТЕРАТУРA:

  • Biljana Andjelic, Radojica Djokovic, Marko Cincovic, Snežana Bogosavljevic-Boškovic, Milun Petrovic, Jelena Mladenovic, Aleksandar Cukic (2022). Relationships between Milk and Blood Biochemical Parameters and Metabolic Status in Dairy Cows during Lactation. Metabolites 2022,12 (8),733; https://doi.org/10.3390/metabo12080733
  • Antunović, Z., Mioč, B., Klir, Z., Šalavardić, K., Širić, I., Držaić, D., Jukić Grbavac, M., Novoselec J. (2022). Correlation between milk compositionand selected blood indicators of liver function in ewes during lactation. Mljekarstvo 72 (1).  3-10. https://doi.org/10.15567/mljekarstvo.2022.0101.
  • Babaei, H., Mansuori-Najand, L., Molaei, M.M., Kheradmand, A., Sharifan, M. (2007): Assessment of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase activities in cow’s milk as an indicator of subclinical mastitis. Veterinary Research Communicationes 31, 419–425. https://doi.org/10.1007/s11259-007-3593-x.
  • Benedet, A., Manuelian, C.L., Zidi, A., Penasa, M., De Marchi, M.(2019):  β-hydroxybutyrate concentration in blood and milk and its associations with cow performance.Animal, 13, (8),1676–1689. DOI: https://doi.org/10.1017/S175173111900034X]
  • Bobe, G., Young, J.W., Beitz, D.C. (2004): Pathology, etiology, prevention, treatment of fatty liver in dairy cows. Journal of  Dairy Science, 87, 3105–3124. https://doi.org/10.3168/jds.S0022-0302(04)73446-3
  • Brunner, N., Groeger, S., Raposo, J.C., Bruckmaier, R.M., Gross, J.J. (2019): Prevalence of subclinical ketosis and production diseases in dairy cows in Central and South America, Africa, Asia, Australia, New Zealand, and Eastern Europe. Translation Animal Science, 3, 19–27., DOI:10.1093/tas/txy102.
  • Chavarria, I., Ángel-García, O., Alvarado, A., Contreras, V., Carrillo, D., Macías-Cruz, U., Avendaño-Reyes, L., Mellado, M. (2025): A time series analysis of monthly Holstein milk yield and composition in a hot environment. Mljekarstvo,75(1), 52-61., DOI: 10.15567/mljekarstvo.2025.0105.
  • Churakov, M., Karlsson, J., Edvarsson-Rasmussen, A., Holtenius K. (2021): Milk fatty acids as indicators of negative energy balance of dairy cows in early lactation. Animal, DOI:15:100253.
  • Chladek ,G.,Machal L.(2004): Blood plasma urea concentration and its relationship with milk production parameters in czech pied cows. Journal of Central Europe Agriculture 5.337–346.
  • Civelek, T., Aydim, I., Cingi, C.C., Yilmaz, O., Kabu, M. (2011): Serum non-esterified fatty acids and beta-hydroxybutyrate in dairy cows with retainedplacenta. Pakistan Veterinary Journal, 31(4), 341-344. ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE).
  • Dann, H.M., Morin, D.E., Murphy, M.R., Bollerog, A., Drackely, J.K. (2005): Prepartum intake, postpartum induction of ketosis, and periparturient disorders affect the metabolic status of dairy cows. Journal of Dairy Science, 88.3249–3264.  DOI: 10.3168/jds.S0022-0302(05)73008-3.
  • Drackley, J.K., Dann, H.M., Douglas, G.N., Janovick-Gurtzky, N.A., Lutherland, N.B., Underwood, J.P.,Loor, .J. (2005): Physiological and pathological adaptations in dairy cows that may increase susceptibility to periparturient diseases and disorders. Italian Journal of  Animal Science, 4. 323–344. DOI:10.4081/ijas.2005.323.
  • Djokovic, R. Ilic, Z., Kurcubic, V., Petrovic, M., Doskovic, D. (2011): Functional and morphological state of the liver in Simmental dairy cows during transitional period. Revue de Médecine Vétérinaire,162.574–579.
  • Djokovic, R. Cincovic, M., Belic, B., Toholj, B., Davidov, I., Hristovska, T. (2015): Relationship between blood metabolic hormones, metabolites and energy balance in Simmental dairy cows during peripartum period and lactation. Pakistan Veterinary Journal, 35. 163–167. ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE).
  • Djoković, R., Cincovic, M., Ilic, Z., Kurćubic, V., Andjelic, B., Petrović, M., Lalic, N., Jasovic, B. (2019): Relationships between contents of biochemical metabolites in blood and milk in dairy cows during transition and mid lactation. International Journal of Applaied Reserch in Veterinary Medicine,  17, (1). 1-9. ISSN: 1542-2666.
  • Djokovic, R., Cincovic, M., Andjelic, B., Petrovic, M.., Cukic, A. (2024): Associations between milk and blood biochemical parameters and metabolic status in dairy cows during lactation. Proceedings, Keynote Lectures, Oral and Poster Presentations. The 32nd World Buiatrics Congress, May 20 to 24, Cancun, Quintana Roo, Mexico, 823-824.
  • Eicher, R. (2004): Evaluation of the metabolic and nutritional situation in dairy herds: Diagnostic use of milk components. Proceedings of the World Buiatrics Congress, 11–16, July, 2004, Quebec, Canada.
  • Ghadaa, E.M. (2014): Investigation of some enzymes level in blood and milk serum in two stages of milk yield dairy cows at Assiut city. Assiut Vetetinary Medicine Journal, 60. 110–120. DOI:10.21608/AVMJ.2014.170982.
  • Gonzalez, F.D., Murino, R., Pereira, V., Campos, R., Benedito, J.L. (2011): Relationship among blood indicators of lipomobilization and hepatic function during early lactation in high yielding dairy cows. Journal of Veterinary Science, 12. 251–255. DOI: 10.4142/jvs.2011.12.3.251
  • Hamann, J., Kromker, V. (1997): Potential of specific milk composition variables for cow health management.  Livestock  Production  Science, 48.201–208.
  • Henao-Velasquez, A.P., Munera-Bedoya, O.D., Herrera, A.C., Aqudelo-Trujillo, J.H., Cenon –Munoz, M.F. (2014). Lactose and milk urea nitrogen, fluctuations during lactation in Holstein cows. Revista Brasilia Zootecnica 43.479:484.  https://doi.org/10.1590/S1516-35982014000900004
  • Jilek, F., Rehak, D., Volek, J., Stipkova, M., Nemcova, E., Fiedlerova, M., Rajmon, A.R., Svetskova, D. (2006): Effect of herd, parity stage of lactation and milk yield on urea concentration in milk. Czech Journal of Animal Science, 51. 510–517. DOI: 10.17221/3971-CJAS
  • Jorritsma, R.H., Jorritsma, Y.H., Schukken, P.C., Bartlett, T., Wensing, T., Wenting, G. (2001): Prevalence and indicators of postpartum fatty infiltration of the liver in nine commercial dairy herds in the Netherlands. Livestock Production Science, 68. 53–60. DOI 10.1016/so301-6226(00)00208-6.
  • Jozwik, A., Strzalkowska, N., Bagnicka, E., Grzybek, W., Krzyzewski, J., Polowska, E., Kolataj, A, Horbanczuk, J.O. (2012): Relationship between milk yield, stage of lactation, and some blood serum metabolic parameters of dairy cows. Czech Journal of Animal Science, 57. 353–360. DOI: 10.17221/6270-CJAS
  • Katsoulos, P.D., Christodoulopoulos, G., Minas, A., Karatzia, M.A., Pourliotis, K., Kritas, S.K. (2010): The role of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase in the diagnosis of subclinical intramammary infections in dairy sheep and goats. Journal of Dairy Research, 77.107–111. doi: 10.1017/S0022029909990410.
  • Le Blanc, S.J. (2010): Monitoring metabolic health of dairy cattle in transition period. Journal of Reproduction and Development, 56 (Suppl1). 29–35.  DOI: 10.1262/jrd.1056s29
  • Liu, P., He, B.X., Yang, X.L., Hou, X.L., Han, J.B., Han, Y.H., Nie, P., Deng, H.F., Du, X.H. (2012): Bioactivity evaluation of certain hepatic enzymes in blood plasma and milk of Holstein cows. Pakistan Veterinary Journal, 32.601–604. ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE).
  • Liu, P., Hou, L.X., Nie, P., Aahan, H.Y., Hoang, F.Y., Zoun, X.Z., Deng, F.H., Song, P., Li, M., Xian,g H.B. (2013): Dynamic Monitoring of ALT and correlation analysis in blood plasma and milk of Holstein cows. Agriculture Journal, 8.51–55
  • Liu, Q., Wang, C., Guo, G., Huo, W.J., Zhang, S.L., Pei, C.X., Zhang, Y.L., Wang, H..
    (2018): Effects of branched-chain volatile fatty acids on lactation performance and mRNA expression of genes related to fatty acid synthesis in mammary gland of dairy cows. Animal, 12.2071–2079. https://doi.org/10.1017/S1751731118000113.
  • Mäntysaari, P., Mäntysaari, E.A., Kokkonen, T., Mehtiö, T., S. Kajava, S., Grelet, C., Lidauer, P., Lidauer, M.H. (2019): Body and milk traits as indicators of dairy cow energy status in early lactation. Journal of Dairy Science, 102,7904–7916. DOI: 10.3168/jds.2018-15792
  • Mordak, R., Kupczyński, R., Kuczaj, M., Niżański, W. (2020): Analysis of correlations between selected blood markers of liver function and milk composition in cows during late lactation period. Annals of Animal Science, 20(3). 871–886. DOI:10.2478/aoas-2020-0020.
  • Nozad, S., Ramin, A.G., Moghadam, G., Rezaei, S.A., Babapour, A., Ramin S. (2007): Relationship between blood urea, protein, creatinine, triglycerides and macro-mineral concentrations with the quality and quantity of milk in dairy Holstein cows. Veterinary Research Forum, 3. 55–59. PMID: 25653747
  • Nozad, S., Ramin, A.G., Moghadam, G. (2011): Diurnal variations in milk, urea, protein and lactose concentrations in Holstein dairy cows. Acta Veterinaria, 61. 3–12. https://doi.org/10.2298/AVB1101003N
  • NRC, National Research Council. (2001): Nutrient requirements of dairy cattle. 7th ed. National Academy Press, Washington, D. C., USA.
  • Oetzel, G.R. 2004: Monitoring and testing dairy herds for metabolic disease. Veterinary Clinic of  North America: Food Animal Practice, 20. 651–674. DOI: 10.1016/j.cvfa.2004.06.006
  • Ospina, P.A., Nydam, D.V., Stokol, T., Overton, TR. (2010): Associations of elevated nonesterified fatty acids and β-hydroxybutyrate concentrations with early lactation reproductive performance and milk production in transition dairy cattle in the northeastern United States. Journal of Dairy Science, 93. 1596–1603. https://doi.org/10.3168/jds.2009-2852
  • Pires, J.A.A., Larsen, T., Leroux, C. (2022): Milk metabolites and fatty acids as noninvasive biomarkers of metabolic status and energy balance in early-lactation cows. Journal of Dairy Science, 105.201–220. DOI: 10.3168/jds.2021-20465
  • Radostits, O.M., Blood, D.C., Gay, C.C., Hinchcliff, K.W. (2000):Veterinary Medicine, A Textbook of the Diseases of Cattle, Sheep, Pigs, Goats and Horses. Ninth Edition W.B.Saunders Company Ltd London, New York, Philadelphia, San Francisco, St. Louis, Sydney.
  • Sevinc, M., Basoglu, A., Guzulbekta, H. (2003): Lipid and lipoprotein levels in dairy cows with fatty liver. Turkish Journal of Veterinary and  Animal Science, 27. 295–299. . Available at: https://journals.tubitak.gov.tr/veterinary/vol27/iss2/2
  • Stang, B.D, Bertics, S.J, Grummer,  R.R, Armentanol, E.(1998): Effect of long chain fatty acids on triglycerides accumulation, gluconeogenesis and ureogenesis in bovine hepatocytes. Journal of Dairy Science, 81.728–739. DOI: 10.3168/jds.S0022-0302(98)75629-2
  • Stojevic, Z., Pirsljin, J., Milinkovic-Tur, S., Zdwlar-Tuk, M., Ljubic, B.B. (2005): Activities of AST, ALT and GGT in clinically healthy dairy cows during lactation and in the dry period. Veterinarski  Arhiv,75.67–73. https://hrcak.srce.hr/67059
  • Stolcova, M., Rehak, D., Barton, L., Rajmon, R.(2020): Blood biochemical parameters measured during the periparturient period in cows of Holstein and Fleckvieh breeds differing in production purpose. Czeck Journal of Animal Science, 65(5).172–181. DOI: 10.17221/99/2020-CJAS
  • Tinkovicova-Lackova, P., Maskalova, M., Vajda, V.(2019): Evaluation of the milk urea content in relation to milk production and composition in dairy cows. Acta Veterinaria Brno, 88. 277–285. https://doi.org/10.2754/avb201988030277
  • Westwood, C.T, Lean, I.J, Kellaway, R.C. (1998): Indications and implications for testing of milk urea in dairy cattle: A Quantitative review. Part 1. Dietary protein sources and metabolism. New Zaeland Veterinary Journal, 46. 87–96. DOI: 10.1080./00480169.1998.36066.
  • Whitaker, D.A., Macrae, A.I., Burrough, E. (2005): Nutrition, fertility and dairy herd productivity. Cattle Practice, 13. 27–32.
  • Woolpert, M.E.,  Dann, H.M., Cotanch, K.W., Melilli, C., Chase, L.E., R. J. Grant, R.J.. D Barbano, D.M. (2016): Management, nutrition, and lactation performance are related to bulk tank milk de novo fatty acid concentration on northeastern US dairy farms. Journal of Dairy Science, 99.8486–8497. DOI: 10.3168/jds.2016-10998