Electronic Journal of Polish Agricultural Universities (EJPAU) founded by all Polish Agriculture Universities presents original papers and review articles relevant to all aspects of agricultural sciences. It is target for persons working both in science and industry,regulatory agencies or teaching in agricultural sector. Covered by IFIS Publishing (Food Science and Technology Abstracts), ELSEVIER Science - Food Science and Technology Program, CAS USA (Chemical Abstracts), CABI Publishing UK and ALPSP (Association of Learned and Professional Society Publisher - full membership). Presented in the Master List of Thomson ISI.
Volume 14
Issue 2
Veterinary Medicine
Available Online: http://www.ejpau.media.pl/volume14/issue2/art-07.html


Robert Kupczyński1, Rafa³ Bodarski2, Maciej Adamski3
1 Department of Environmental Hygiene and Animal Welfare, Wroc³aw University of Environmental and Life Sciences, Poland
2 Department of Animal Nutrition and Feed Science, Wroc³aw University of Environmental and Life Sciences, Poland
3 Institute of Animal Breeding, Wroc³aw University of Environmental and Life Sciences, Poland



The aim of the study was an assessment of an influence of glycerin applied to the TMR dose on carbohydrate-lipid transformations, chosen enzymes activity and milk yield of Polish Holstein-Friesian cows in periparturient period. 24 multiparous cows were included in the research. Glycerin in a dose of 300 ml /day was applied to the TMR dose of the experimental cows from 7 day a.p. to 3 week p.p. Biochemical tests of blood were conducted using Pentra 400 analyzer of HORIBA ABX Company. The concentration of glucose, β-betahydroxybutyrate acid, unsaturated fatty acid, triglycerides, total cholesterol, AST, ALP, GGT enzymes activity as well as total bilirubin was determined in blood serum. The long-term feeding of glycerin in periparturient period had an antiketogenic effect limiting glucose level decrease in blood, and limited an intensification of lipolysis at the beginning of lactation. The lower increase of AST activity and total bilirubin concentration was affirmed at 3 week p.p., after the use of glycerin. The lower decrease in condition was noted in experimental cows, that did not influence statistically on milk yield.

Key words: cows, periparturient period, glycerin, blood, metabolic profile.


A sudden growth in energy and essential nutrients requirements in a proper development of foetus, production of colostrum and milk is observed in periparturient period and at the beginning of lactation, concurrently with hormonal changes causing an organism subordination to lactation process [1,6,13]. The energy deficiency at the beginning of lactation is compensated by a growth of reserve fat lipolysis, that results in an increased non-esterified fatty acids (NEFA) content in blood [7,8,24]. NEFA are metabolized in the liver to triglycerides (TG), or they are transformed to ketonic compounds [11,19]. An intensified or elongated lipolysis leads to fatty liver, while large amounts of acetyl-CoA formed during NEFA β-oxydation process intensify the hyperketogenesis [22,23].

Numerous research concerning an application of glucogenic precursors in a transition period, mainly propylene glycol as antiketogenic compound that also decreases the risk of fatty liver, have been conducted so far [3,10,12,16,20,22]. Considerably smaller amount of research concerned the use of glycerol at the same purpose. An interest in glycerol application in ruminants results also from biodiesel production development, where glycerin is a waste product.

Goff and Horst [9] demonstrated that feeding with glycerin per os is effective in ketosis prevention in periparturient cows. Feeding the cows with glycerol caused an increase in glucose concentration in blood [9,18], and a decrease in free fatty acids level [18]. Similar results were obtained by Chung et. al. [4]. A lack of antiketogenic effect of glycerin addition to TMR doses was demonstrated in another study [2,5]. The present study aimed at an assessment of metabolic effects resulting form glycerin application in TMR dose of Polish Holstein-Friesian breed in periparturient period.


The study included dairy cows of Polish Holstein-Friesian breed coming from a farm of a mean yield on a level of 9000 kg of milk in 305 d lactation. The animals in periparturient period were kept in a nursery, on deep litter in a theathering system. Feeding was based on TMR (Total Mixed Ration) diets calculated according to DLG system. The two diets were used, i.e. for close-up period (2–3 weeks prepartum) and for lactating cows (Table 1).

Table 1. Diet (TMR) for cows during an experiment (kg/d)


2–3 weeks prepartum


Corn silage



Partially wilted lucerne silage



Pressed pulp silage


Fresh spent grain



Breeder mash



Extracted soybean meal



Corn grain silage



Protected fat


Wheat straw



Mineral mixture for dry period


Productive mineral mixture


Forage chalk


Acid sodium carbonate 


Nutritive value

Dry matter kg/d






nCP g/ d



(CP-nCP)/6.25 g



Twenty four cows (multiparous ones) were chosen for the experiment on the basis of analogues. Clinically healthy cows, aged 4–7 years, were divided into 2 groups (n=12): control group fed with TMR diet, and experimental one which received 300 ml of glycerin top-dressed with TMR.

The cows were given the glycerin in a liquid form (99.7% content of 1,2,3-propantriol) once a day during the morning feeding (Over, Poland). The preparation was top-dressed and hand-mixed into the upper one third of the daily ration. The preparation was given starting one week before an expecting parturition date, continuing to 21st day of lactation.

Blood for the experiment was collected from an external jugular vein (v. jugularis externa) at the morning, before feeding, one week before expected parturition date and at 7th and 21st day post partum. Laboratory analyses were done using Pentra 400 biochemical analyzer Company (Horiba ABX Diagnosis, Montpellier Cedex, France). The analyses included biochemical parameters that determine the most important directions of carbohydrate-lipid transformations and are markers of liver functioning, i.e.:

The data concerning milk yield was obtained from milking system of milking parlour (Westfalia Company). The condition of cows that were included in the experiment was determined using BCS method (1–5 points, 0.25 point range) at the first day of the experiment, and in 1st, 3rd and 10th week of lactation.

Basic statistical parameters of the results (means, standard deviations) in individual groups were worked out using the Statgraphics ver. 5.0 software. The significance of differences between the experimental factors was determined by Duncan's test.


The condition of cows in both groups before parturition was even (Table 2). A successive, significant decrease (P<0.01) in the condition, until 10th week of lactation, was observed after the parturition. It was 0.8 point in the control group, and 0.67 point in the experimental one. No statistical differences in milk yield of analysed cows were observed (Fig. 1). Mean daily milk yield of the cows from the control group was slightly lower (37.94 kg) when compared to the cows from the experimental group (39.17 kg) until the 10th week of lactation.

Table 2. Estimation of cows' condition score based on the BCS method (points), ( ± s)


Estimation term

1 week a.p.

Period of lactation

1 week

3 week

10 week

Control (n = 12)





(n = 12)





1 cows were fed 300 ml/d of glycerin added to TMR from week before calving until 21 days after calving
a – statistically significant differences between prepartum and postpartum period, p≤0.05
A, B – statistically significant differences between prepartum and postpartum period, p≤0.01
* – statistically significant differences between groups p≤0.05

Fig. 1. Milk yield of cows in the first weeks of lactation

The values of analysed carbohydrate-lipid transformation parameters did not reveal any considerable differences in both groups at the beginning of the experiment (Table 3). After parturition, in 3rd week of lactation, the significant decrease in glucose concentration in both groups (P≤0.01) was noted. The higher glucose level was observed in cows fed with glycerin (Table 3). The beginning of lactation in a significant manner affected the concentration of BHBA and NEFA in blood serum. The more intensified ketogenesis and lipolysis were observed in the case of control cows. Lower content of BHBA (P≤0.05) was noted in 3rd week postpartum in the group fed with glycerol when compared to the control one. The content of triglycerides and  total cholesterol decreased after parturition in control cows (P≤0.05), that was not observed in the experimental group (Table 3). In the 3rd week of lactation, the experimental cows were characterized by the higher level of these parameters (P≤0.01) when compared to the control ones.

Table 3. Means of serum blood metabolites concentrations pre- and postpartum,
(  ± s)





n = 12

n = 12

Glucose (mmol/)







BHBA (mmol/l)







NEFA (mmol/l)







Triglycerides (mmol/l)







Total cholesterol (mmol/l)







1 cows were fed 300 ml/d of glycerin added to TMR from week (3-10 d) before calving until 21 days after calving
* – statistically significant differences between prepartum and postpartum period p≤0.05
** – statistically significant differences between prepartum and postpartum period p≤0.01
a, b – statistically significant differences between groups, p≤0.05
A, B – statistically significant differences between groups, p≤0.01

Table 4. Enzymatic activities and total bilirubin content in blood serum (  ± s)





n = 12

n = 12

AST (U/l)







ALT (U/l)







GGT (U/l)







Total bilirubin (µmol/l)







Explanations: like in Table 3

Before the parturition, AST activity was higher (P≤0.05) in control cows when compared to the experimental group (Table 4). An increase in AST activity during postpartum period was observed in control (up to 97.28 U/L) and experimental cows (63.99 U/L). The changes in ALT activity were small, however its increased activity after parturition was confirmed statistically (P≤0.05). There were no significant changes in GGT activity during experimental period. Total bilirubin concentration in blood serum increased in both groups after parturition (Table 4). The experimental treatment caused the minor increase in total bilirubin concentration in blood serum. Differences between the groups were significant statistically (P≤0.05).


More research concerning the application of propylene glycol when compared to those on glycerin have been conducted so far. An activity of propylene glycol is based on a mechanism causing a decrease in NEFA and ketonic compounds content in blood, and an increase in glucose and insulin concentration [3,10,22]. Feeding with propylene glycol causes an increase in L-lactates and propionates supply for gluconeogenesis [14]. The hyperglycemic and hyperinsulinaemic effects of glycol are most likely caused by insulin resistance induced by an increased glycol and propanol concentrations, and a decreased ratio of ketogenic and glucogenic metabolites in arterial blood plasma [14]. Grummer [11] points an antilipolytic effect of that compound. Propylene glycol used in a restrictive feeding of heifers was more effective when fed per os, than when it was given as a part of concentrate or added to TMR dose [3]. However, oral drenching and esophageal pumping present tremendous stress to dairy cows, especially in the early postpartum period. Glycerin in a liquid form added to TMR dose was used in the present study.

The condition decrease at the beginning of lactation up to about 2.75 points was not connected with an increase in NEFA and BHBA in blood [13]. In the present study on high-yielding cows, a slightly intensified lipolysis and ketogenesis were observed in 3rd week of lactation, especially in the control group. The observed fall of the condition can be recognized as a physiological one with given milk yield level. Short-term application of glycerin to water in periparturient period (7 days prepartum to 7 days postpartum) did not affect the condition of cows [17]. The increased fodder intake was observed in the present study, however, it did not influence energy balance nor milk yield. Glycerin used in a form of powder for the first three weeks of lactation (162.5g/d only), did also not influence the milk yield of cows [4]. The tendency towards the higher milk yield at glycerin supplemented cows during 6 weeks of lactation (52 vs. 46 kg/d) after the supplementation period suggested a potential benefit of dry glycerin on subsequent milk production [19]. An application of glycerin in a higher dose (300-500 g/d) to cows from the 3rd week prepartum to 70th day postpartum, caused a significant yield increase (P<0.01) in the first 10 weeks of lactation [2]. Higher yield in the case of cows that were given glycerin, that was willingly consumed mixed with TMR dose, was also observed in the present study. That observation is consistent with the study by Spörndly and Åsberg [21], who compared 25 kinds of fodders with respect to willingness of their consumption by cows. Barley with glycerol addition (10%) was placed at the 5th position.

According to Chung et al. [4], an application of powdered glycerin in a transition period influences an improvement of energetic status, causing an increase in glucose concentration, and a decrease in BHBA concentration in blood, and ketonic compounds in urine. In another study, glycerol given during first 2 weeks after parturition (per os) caused a decrease in glucagon, NEFA, and BHBA content in blood, exhibiting a glucogenic effect at the same time (18). The mentioned study demonstrated more intense, profitable influence of joint injection of glucagon and glycerol on metabolism. The present study demonstrated significantly lower (P≤0.05) content of BHBA in the 3rd week postpartum in experimental cows. Goff and Horst [9] believe that antiketogenic influence of glycerin applied per os depends on its dose. The newest study analysed glycerin application in drinking water in amount of 20 g/l from 7th day prepartum to 7th d postpartum [17]. The study did not demonstrate any glucogenic effect in such a short time, however the level of BHBA decreased after parturition. DeFrain et al. [5] noted a decrease in glucose content in blood, and an  increase in BHBA concentration while glycerol was supplemented to TMR doses. The study demonstrated that glycerin increased an overall content of volatile fatty acids and decreased the ratio of acetates to propionates. Glycerin used in the present study was pure, as opposed to that used by DeFrain et al. [5] with glycerol content on a level of 80.2%.

Glycerin used in the present study caused smaller increase in AST activity and total bilirubin concentration in blood serum. Using energetic supplements in a transition period, no influence on AST and ALT activity was observed [1], while an application of propylene glycol to TMR doses for cows in periparturient period caused a decrease in AST activity in blood [16]. An assessment of enzymes activity was not conducted in any other study concerning glycerin application [4,17].

According to Dracley et al. [7] glucose deficiency on a level of 500 g/d is characteristic for an early lactation stage of cows. It may be limited by an application of glucogenic precursors in a diet. Glycerin may be such an additive, as  it was demonstrated in results of another study [4,18] and those presented in the present paper.


It should be stated that intensified lipolysis in high-yielding cows of Holstein-Friesian breed in the first stage of lactation is a physiological phenomenon, even with balanced nutritional dose and supplementation with a glucoplastic compound. Long-term glycerin application to cows contributed in an improved milk field when compared to control cows (39.17 kg vs 37.94 kg). Long-term top-dress application of glycerin in TMR in periparturient period may result in an antiketogenic effect, profitably influencing an activity of enzymes analysed.


  1. Ballard C.S., Mandebvu, P., Sniffen, C.J., Emanuele, S.M., Carter, M.P., 2001. Effect of feeding an energy supplement to dairy cows pre- and postpartum on intake, milk yield, and incidence of ketosis. Anim. Feed Sci. Technol., 93, 55–69.

  2. Bodarski, R., Wertelecki, T., Bommer, F., Gosiewski, S., 2005. The changes of metabolic status and lactation performance in dairy cows under feeding TMR with glycerin (glycerol) supplement at periparturient period. EJPAU 8(4), #22, http://www.ejpau.media.pl/volume8/issue4/art-22.html.

  3. Christensen, J.O., Grummer, R.R., Rasmussen, F.E., Bertics, S.J., 1997. Effect of method of delivery of propylene glycol on plasma metabolites of feed-restricted cattle. J. Dairy Sci., 80, 563–568.

  4. Chung, Y.H., Rico, D.E., Martinez, C.M., Cassidy, T.W., Noirot, V., Ames, A., Varga, G. A., 2007. Effects of feeding dry glycerin to early postpartum holstein dairy cows on lactational performance and metabolic profiles. J. Dairy Sci., 90, 5682–5691.

  5. DeFrain, J.M., Hippen, A.R., Kalscheur, K.F., Jardon, P.W., 2004. Feeding glycerol to transition dairy cows: effects on blood metabolites and lactation performance. J. Dairy Sci., 87, 4195–4206.

  6. Doepel L., Lapierre, H., Kennelly, J.J., 2002. Peripartum performance and metabolism of dairy cows in response to prepartum energy and protein intake J. Dairy Sci., 85, 2315–2334.

  7. Drackley, J.K., Overton, T.R., Douglas G. N., 2001. Adaptations of glucose and long-chain fatty acid metabolism in liver of dairy cows during the periparturient period. J. Dairy Sci., 84, 100–112.

  8. Duffield, T.F., 2000. Subclinical ketosis in lactating dairy cattle: Metabolic disorders of ruminants. Vet. Clin. North Am. Food Anim. Pract., 16, 231–253.

  9. Goff, J.P., Horst, R.L., 2001. Oral glycerol as an aid in the treatment of ketosis/fatty liver complex. J. Dairy Sci., 84, Suppl. 1, 153.

  10. Grummer, R.R., Winkler, J.C, Bertics, S.J., Studer, V.A., 1994. Effect of propylene glycol dosage during feed restriction on metabolites in blood of prepartum Holstein heifers. J. Dairy Sci., 77, 3618–3623.

  11. Grummer, RR., 2008. Nutritional and management strategies for the prevention of fatty liver in dairy cattle. Vet. Journal., 176, 10–20.

  12. Hachenberg, S., Weinkauf, C., Hiss, S., Sauerwein, H., 2007. Evaluation of classification modes potentially suitable to identify metabolic stress in healthy dairy cows during the peripartal period. J. Anim. Sci., 85, 1923–1932.

  13. Hoedemaker, M., Prange, D., Zerbe, H., Frank, J., Daxebberger, A., Meyer, H.H.D., 2004. Peripartal propylene glycol supplementation and metabolism, animal health, fertility and production in dairy cows. J. Dairy Sci., 87, 2136–2145.

  14. Kristensen, N.B., Raun, B.M.L., 2007. Ruminal and intermediary metabolism of propylene glycol in lactating holstein cows. J. Dairy Sci., 90, 4707–4717.

  15. Kupczyński, R., Janeczek, W., Pogoda-Sewerniak, K., 2005. Studies on the use of different doses of propylene glycol in dairy cows during the periparturient period. Medycyna Wet., 61, 194–199.

  16. Miku³a, R., Nowak, W., Ja¶kowski, J.M., Maękowiak, P., Pruszyńska, E., W³odarek, J., 2008. Effects of propylene glycol supplementation on blood biochemical parameters in dairy cows. Bull. Vet. Inst. Pulawy, 52, 461–466.

  17. Osborne, V.R., Odongo, N.E., Cant, J.P., Swanson, K.C., McBride, B.W., 2009. Effects of supplementing glycerol and soybean oil in drinking water on feed and water intake, energy balance, and production performance of periparturient dairy cows. J. Dairy Sci., 92, 698–707.

  18. Osman, M.A., Allen, P.S., Mehyar, N.A., Bobe, G., Coetzee, J.F., Koehler, K.J., Beitz, D.C., 2008. Acute metabolic responses of postpartal dairy cows to subcutaneous glucagon injections, oral glycerol, or both. J. Dairy Sci., 91, 3311–3322.

  19. Rukkwamsuk, T., Rungruang, S., Choothesa, A., Wensing, T., 2005. Effect of propylene glycol on fatty liver development and hepatic fructose 1,6 bisphosphatase activity in periparturient dairy cows. Livest. Product. Sci., 95, 95–102.

  20. Rukkwamsuk, T., Wensing, T., Geelen, M.J.H., 1999. Effect of overfeeding during the dry period on the rate of esterification in adipose tissue of dairy cows during the periparturient period. J. Dairy Sci., 82, 1164–1169.

  21. Spörndly, E., Åsberg, T., 2006. Eating rate and preference of different concentrate components for cattle. J. Dairy Sci., 89, 2188–2199.

  22. Studer ,V.A., Grummer, R.R., Bertics, S.J., Reynolds, C.K., 1993. Effect of prepartum propylene glycol administration on periparturient fatty liver in dairy cows. J. Dairy Sci., 76, 2931–2939.

  23. Suriyasathaporn, W., Heuer, C., Noordhuizen-Stassen, E.N., Schukken, Y.H., 2000. Hyperketonemia and the impairment of udder defense: a review. Vet. Res., 31, 397–412.

  24. Vazquez-Anon, M., Bertics, S., Luck, M., Grummer, R.R., Pinheiro, J., 1994. Peripartum liver triglyceride and plasma metabolites in dairy cows. J. Dairy Sci., 77, 1521–1528.


Accepted for print: 2.02.2011

Robert Kupczyński
Department of Environmental Hygiene and Animal Welfare,
Wroc³aw University of Environmental and Life Sciences, Poland
Che³mońskiego 38 C, 51-630 Wroc³aw, Poland
Phone: (+48 71) 320 59 41
email: robert.kupczynski@up.wroc.pl

Rafa³ Bodarski
Department of Animal Nutrition and Feed Science,
Wroc³aw University of Environmental and Life Sciences, Poland
Che³mońskiego 38 C, 51-630 Wroc³aw, Poland
email: bodarski@zoo.ar.wroc.pl

Maciej Adamski
Institute of Animal Breeding,
Wroc³aw University of Environmental and Life
Sciences, Poland
Che³mońskiego 38C, 51-630 Wroc³aw, Poland

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