The Application of Near-infrared spectroscopy technique for measuring plasma Beta-hydroxybutyrate and non-esterified fatty acid concentration and subclinical ketosis screening test in dairy cow

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theerakul Nilnont
Rittichai pilachai
Pranpreya Todtong
Saranya Workhwa


Introduction: Ketosis is one of the metabolic diseases that causes economic loses in dairy industry. The evaluation of BHB and NEFA is a specific test for subclinical ketosis which shows the higher blood level of beta-hydroxybutyrate (BHB) and non-esterified fatty acid (NEFA). There have been applied the Near-infrared spectroscopy (NIRS) technique for detecting subclinical ketosis, however the studies of this technique are needed for supporting the accuracy of the analysis and screening tests.

Objectives: The aims of this study were to apply the NIRS technique to measure plasma BHB and NEFA of dairy cow by generating the appropriate equation to predict the relationship between the laboratory BHB and NEFA results and the NIRS results, and to investigate the prevalence of subclinical ketosis in dairy raising area, Udon Thani province.

Materials and methods: blood and plasma samples of 102 post-partum dairy cows (5-60 DIM) were collected. The plasma samples were separated and divided into 2 parts, for laboratory BHB and NEFA analysis and NIRS analysis by using FT-NIR and DLP-NIR. The appropriate equation was generated to predict the BHB and NEFA level.    

Results: The prevalence of subclinical ketosis in pots-partum dairy cows was 5.88% (6/102). The statistical analysis to generate the equation to predict plasma BHB and NEFA level, by using FT-NIR และ DLP-NIR, demonstrated the R value were 0.98, 0.90 and 0.98, 0.84, respectively.

Conclusions: The results of this study showed the moderate relationship between the laboratory and the NIRS BHB and NEFA level and was able to apply in the research. The further studies and data collections is necessary for improve the accuracy of blood chemistry analysis by using NIRS technique.


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Adewuyi AA, Gruys E, Van Eerdenburg FJCM, 2005. Non esterifiedfatty acids (NEFA) in dairy cattle. A review, Vet Q 27(3), 117-126.

Andersson L, 1988. Subclinical ketosis in dairy cows. Vet Clin North Am Food Anim Pract 4(2), 233-51.

Asl AN, Nazifi S, Ghasrodashti AR, Olyaee A, 2011. Prevalence of subclinical ketosis in dairy cattle in the Southwestern Iran and detection of cutoff point for NEFA and glucose concentrations for diagnosis of subclinical ketosis. Prev Vet Med 100(1), 38-43.

Baird GD, 1982. Primary ketosis in the high-producing dairy cow: clinical and subclinical disorders, treatment, prevention, and outlook. J Dairy Sci 65(1), 1-10.

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. Transl. Anim. Sci 3, 19-27.

Denis-Robichaud J, Dubuc J, Lefebvre, DesCôteaux L, 2014. Accuracy of milk ketone bodies from flow-injection analysis for the diagnosis of hyperketonemia in dairy. J Dairy Sci 97, 3364–3370.

De Roos A.P, van den Bijgaart J, Horlyk G, De Jong, 2007. Screening for subclinical ketosis in dairy cattle by Fourier transform infrared spectrometry. J Dairy Sci 90, 1761-1766.

Dervishi E, Plastow G, Hoff B, Colazo M, 2021. Common and specific mineral and metabolic features in dairy cows with clinical metritis, hypocalcaemia or ketosis. Res Vet Sci 135, 335-342

Dohoo I, Martin W, Stryhn, H. (2010) Veterinary Epidemiologic Research. VER Inc., Charlottetown.

Dohoo IR, Martin SW, 1984. Subclinical Ketosis: Prevalence and Associations with Production and Disease. Can J Comp Med 48, 1-5.

Duffield T, 2000. Subclinical Ketosis in Lactating Dairy Cattle. North Am Food Anim Pract 16(2), 231-253.

Duffield T, Bagg R, 2002. Herd level indicators for the prediction of high-risk dairy herds for subclinical ketosis. In: Proceeding of American Association of Bovine Practitioner, Rome, GA.175-176

Duffield TF, Lissemore KD, McBride BW, Leslie KE, 2009. Impact of hyperketonemia in early lactation dairy cows on health and production. J Dairy Sci 92(2), 571-580.

Enjalbert F, Nicot MC, Bayourthe C, Moncoulon R, 2001. Ketone bodies in milk and blood of dairy cows: Relationship between concentrations and utilization for detection of subclinical ketosis.J. Dairy Sci 84, 583-589.

Gohary K, Overton MW, Von Massow M, LeBlanc SJ, Lissemore KD, Duffield TF, 2016. The cost of a case of subclinical ketosis in Canadian dairy herds. Can Vet J 57(7), 728-32.

Gordon JL, Leblanc SJ, Duffield TF, 2013. Ketosis treatment in lactating dairy cattle. Vet Clin North Am Food Anim Pract 29(2), 433-45.

Herdt TH, 1997.Gastrointestinal physiology and metabolism. Textbook of Veterinary Physiology, 2nd edition. Edited by J. G. Cunningham. WB Sanders Company 1997, 263-380.

Herdt TH, 2000. Influences on the etiology of ketosis and fatty liver. North Am Food Anim Pract 16, 215-230.

Jenkins NT, Peña G, Risco C, Barbosa C, Vieira-Neto, A, Galvão KN, 2015. Utility of inline milk fat and protein ratio to diagnose subclinical ketosis and to assign propylene glycol treatment in lactating dairy cows. Can Vet J 56(8), 850–854.

Jorjong S, van Knegsel ATM, Verwaeren J, Val Lahoz M, Bruckmaier RM, De Baets B, Kemp B , Fievez V, 2014. Milk fatty acids as possible biomarkers to early diagnose elevated concentrations of blood plasma nonesterified fatty acids in dairy cows. J Dairy Sci 97(11), 7054-7064.

LeBlanc S, 2010. Monitoring metabolic health of dairy cattle in the transition period. J Reprod Dev 56, 29-35.

Li Y, Ding H, Liu L, Song Y, Du X, Feng S, Wang X, Li X, Wang Z, Li X, Li J, Wu J, Liu G, 2020. Non-esterified Fatty Acid Induce Dairy Cow Hepatocytes Apoptosis via the Mitochondria-Mediated ROS-JNK/ERK Signaling Pathway. Front Cell Dev Biol28(8), 245.

Liu L, Shen T, Yang W, Yu H, Gao S, Huang B, Xu C, 2020. Ketotic cows display a different serum nonesterified fatty acid composition. J Dairy Res 87(1), 52-55.

Macmillan K, López Helguera I, Behrouzi A, Gobikrushanth M, Hoff B , Colazo MG, 2017. Accuracy of a cow-side test for the diagnosis of hyperketonemia and hypoglycemia in lactating dairy cows. Res Vet Sci 115, 327-331.

Mahrt A, Burfeind O, Heuwieser W, 2015. Evaluation of hyperketonemia risk period and screening protocols for early-lactation dairy cows. J Dairy Sci 8(5), 3110-3119.

McParland S, Berry DP, 2016. The potential of Fourier transform infrared spectroscopy of milk samples to predict energy intake and efficiency in dairy cows. J Dairy Sci 99(5), 4056-4070.

Oetzel GR, 2004. Monitoring and testing dairy herds for metabolic disease. Vet Clin North Am Food Anim Pract 20, 651-674.

Ospina P, Nydam D, Stokol T, Overton T, 2010. Associations of elevated nonesterified fatty acids and beta-hydroxybutyrate concentrations with early lactation reproductive performance and milk production in transition dairy cattle in the northeastern United States. J Dairy Sci 93, 1596-1603.

Ospina P, Jessica M, Overton T, Stokol T, Nydam D, 2013. Using Nonesterified Fatty Acids and β-Hydroxybutyrate Concentrations During the Transition Period for Herd-Level Monitoring of Increased Risk of Disease and Decreased Reproductive and Milking Performance. Vet Clin North Am Food Anim Pract 29, 387-412.

Promkot C, Mansathit J, Wanapat M, 2021. Metabolic Disorders of Transitional Low Production Dairy Cow. J. Agric. Sci. Technol 1(8), 1221-1223.

Ren Z, Wang Y, Liu H, Zhou Z, Cai D, Hu Y, Zong X, Cao S, 2020. Characterization of serum adiponectin and leptin in healthy perinatal dairy cows or cows with ketosis, and their effectson ketosis involved indices. Pol J Vet Sci 23(3), 373-381.

Rukkwamsuk T, Kruip TA, Wensing T, 1999. Relationship between overfeeding and overconditioning in the dry period and the problems of high producing dairy cows during the postparturient period. Vet. Q, 21, 71-77.

Rukkwamsuk T, 2005. Research for improving nutrition and health of dairy cow: in medium scale dairy farm, in: Proceedings of AHAT/BSAS International Conference: Integradting Livestock-Crop Systems to Meet the Challenges of Globalisation, Nov. 14-18, Khon Kaen, Thailand,3, 96-99.

Rutherford AJ, Oikonomou G, Smith RF, 2016. The effect of subclinical ketosis on activity at estrus and reproductive performance in dairy cattle. J Dairy Sci 99(6), 4808-4815.

Simasatitkul N, Supaluckbunlue R, 1998. Ketonuria in Dairy Cows and Correlation between Ketonuria Level and Milk Production, KKU. Vet. J 8(1-2), 19-27.

Sukhontarad P, Yawongsa A, 2020. The Measurement of Serum Beta-hydroxybutyrate Concentrations in Dairy Cows by Near Infrared Spectroscopy. J. Kasetsart Vet 30(2): 61-78.

Tveit B, Lingaas F, Svendsen M, Sjaastad OV, 1992. Etiology of acetonemia in Norwegian cattle.Effect of ketogenic silage, season, energy level, and genetic factors. J Dairy Sci 75(9), 2421-2432.

Suthar VS, Canelas-Raposo J, Deniz A, Heiwieser W, 2013. Prevalence of subclinical ketosis and relationships with postpartum diseases in European dairy cows. J Dairy Sci 96, 2925-2938.

Toledo-Alvarado H, Vazquez AI, de Los Campos G, Tempelman RJ, Bittante G, Cecchinato A, 2018. Diagnosing pregnancy status using infrared spectra and milk composition in dairy cows. J Dairy Sci 101(3), 2496-2505.

Van Knegsel ATM, van der Drift SGA, Horneman M, de Roos APW, Kemp B, Graat EAM, 2010. Ketone body concentration in milk determined by Fourier transform infrared spectroscopy: Value for the detection of hyperketonemia in dairy cows. J Dairy Sci 93, 3065-3069.

Voyvoda H, Erdogan H, 2010. Use of a hand-held meter for detecting subclinical ketosis in dairy cows. Res Vet Sci 89(3), 344-51.

Willams, P, 2007. Near-infrared technologygetting the best out of light. PDK Graqin,Canada. Blackwell Pub.

Wang H, Gao Y, Xia C, Zhang H, Qian W, Cao Y,2016. Pathway analysis of plasma different metabolites for dairy cow ketosis, Ital. J. Anim. Sci, 15(3), 545-551.

Wu ZL, Chen SY, Hu S, Jia X, Wang J, Lai SJ, 2020. Metabolomic and Proteomic Profiles Associated With Ketosis in Dairy Cows. Front Genet 16(11), 551-587.

Xu S, Wu Z, Zou Y, Li S, Cao Z, 2017. Evaluation of a Hand-Held Meter to Detect Subclinical Ketosis in Dairy Cows. J Adv Dairy Res 5(2), 173-178.

Yawongsa A, Sukhontarad P, 2018. Preliminary study on measurement of blood chemistry concentrations in dairy cows serum by near infrared spectroscopy. J Kasetsart Vet 28, 69-87.

Zakian A, Tehrani-Sharif M, Mokhber-Dezfouli MR, Nouri M, Constable PD, 2017. Evaluation of a point-of-care electrochemical meter to detect subclinical ketosis and hypoglycaemia in lactating dairy cows. Aust Vet J 95(4), 123-128.

Zhang H, Wu L, Xu C, 2013. Plasma metabolomic profiling of dairy cows affected with ketosis using gas chromatography/mass spectrometry. BMC Vet Res 9, 186.