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Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 Krsmanović, M. 12
INTRODUCTION
Deviation from the normal values of the inorganic blood
parameter status of cows in early lactation, as well as their
defciency in the diet can lead to subclinical or clinical
manifestations (puerperal paresis, tetany), which have a
negative impact on health and fertility (1, 2, 3, 4, 5). Late
pregnancy and the beginning of lactation present a burden
for the cow’s physiology, as it has increased demands in
calcium, which is necessary for the construction of the fetal
skeleton. Phosphorous represents the second most important
macro-element for bone tissue production. In addition, it
participation in the cellular process of phosphorylation and
energy production, afects acid-base balance and plays a role
in the detoxifcation process (6, 7).
Determination of calcium, magnesium and inorganic
phosphorus in the blood serum is of important diagnostic
value in preventing puerperal paresis and other diseases (8,
9). However, their values can difer in cattle for many rea-
sons, among which is the breed (10). Te values of the above
mentioned macro-minerals in Simmental cows particularly
at diferent production stages are lacking in the literature.
Tis study was aimed at determining and comparing levels
of serum calcium, inorganic phosphorus and magnesium pre-
parturienly, peri-parturiently and during the peak of lactation
in Simmental dairy cattle.
MATERIALS AND METHODS
Animal selection and study design
Te study was conducted in a dairy cattle farm consisted of
150 Simmental cows. In total, 45 clinically healthy cows of
3 diferent productive stages were selected. Te animals of
the 1
st
group (15 cows - group A) were close-up cows, at the
Determination of Macroelement Parameters in Diferent
Productive Stages of Simmental Cows
Krsmanović, M.,
1
Djoković, R.,
1
Giadinis, N.D.,
2
Panousis, N.,
2
Bojkovski, D.,
3
Savić-Stevanović, V.,
4
Vasić, A.,
4
Zdravković, N.,
4
Korica, S.
5
and Bojkovski, J.
4
*
1
Agricultural Faculty, University of Kragujevac, Cara Dušana 34, 32000 Čačak, Serbia.
2
Faculty of Veterinary Medicine, Aristotle University of Tessaloniki, University Campus, 54124 Tesaloniki, Macedonia, Greece.
3
Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia.
4
Faculty of Veterinary Medicine, University of Belgrade, Bul.Oslobodjenja 18, 11000 Belgrade, Serbia.
5
University Union, Faculty for Ecology and Environmental Protection, Cara Dušana 62-64, 11000 Belgrade, Serbia.
*
Corresponding Author: Jovan Bojkovski, DVM. PhD, Associate Professor, Department for Farm Animal Disease, Faculty of Veterinary Medicine,
University of Belgrade, Belgrade, 11.000 Belgrade, Serbia. Tel: +381113615436. Email:bojkovski@vet.bg.ac.rs
ABSTRACT
Te results of inorganic blood parameters are presented in the periparturient period and during lactation of
Simmental dairy cows. Serum calcium and inorganic phosphorus in blood of dairy cows in puerperium were
signifcantly lower (p<0.05) compared to the values in the blood of dairy cows in advanced pregnancy and
during peak of lactation, probably indicating the increased use of these macro-elements by the Simmental
cows’ mammary gland at the early stages of lactation. When the cows were in the periparturient period
serum magnesium levels were signifcantly lower (p<0.05) compared to the values of dairy cows during their
maximal lactation probably indicating the increased use of magnesium by the Simmental dairy cow during
the periparturient period.
Keywords: Simmental cows; Blood Serum; Calcium; Phosphorus; Magnesium.
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 13 Macroelement Parameters of Simmental Cows
last 15 days before the expected parturition. Te animals of
the 2nd group (15 cows - group B) were fresh cows 1 to 15
days after calving, while the 3rd group (15 cows - group C)
consisted of high-production cows, from the 90
th
to 100
th
day
of lactation. All the included cows were in their 3
rd
lactation
and their average milk production was 6,825± 305 liters per
lactation period.
Feeding and housing were in accordance with the in-
tended use of the animals. Diet composition was adjusted to
the requirements of cows during pregnancy and lactation. Te
feeding regime of the cows at diferent production stages is
presented in Table 1.
Cows were judged to be clinically healthy for the duration
of the study.
Table 1: Feeding cows before and after calving
Cows before the calving were
fed daily
Cows after calving and during
lactation consumed daily
3 kg alfalfa hay 1,800 Kg alfalfa hay
Wheat straw 3 kg, 10 kg 15 kg of corn silage
(30% dry weight)
Corn silage (30% dry weight ) 8 kg of alfalfa silage
4 kg alfalfa haylage 4 kg of maze ear silage
(68% dry weight)
2 kg maize ear silage
(68% dry weight)
2 kg of dry sugar beet pulp
0.5 kg dry sugar beet pulp 2 kg exstruded soybeans
1.5 kg of additional mixtures
(30% of total protein)
4.5 kg additional mixtures
(30% of total protein)
Blood sampling
Blood sampling of all the tested cows was conducted once,
by jugular veni-puncture into 20 ml sterile vacuum glass
tubes. All samples were drawn between 10:00-12:00 a.m. Te
samples were placed in ice and were forwarded to the labora-
tory within an hour; sera were separated by centrifugation at
2,500 × g for 15 min. Tereafter the sera were placed in plas-
tic tubes (1.5 ml) (Eppendorf, Germany) and stored at -20°C
until analysed. Te concentration of calcium and magnesium
was determined by atomic absorption spectrophotometry
(AAS), and inorganic phosphorus by a spectrophotometrical
assay kit (Inorganic Phosphorus Assay Kit, Bioo Scientifc
Corporation, USA) using the spectrophotometer, UV-1800
(Shimadzu, Japan). Serum levels of calcium, magnesium and
phosphorus in sera were determined by the standard method
of by atomic absorption spectrophotometry (Serbian SRPS
ISO 6869 2002 using UNICAM 969, USA).
Statistical analysis
Te ANOVA procedure followed by LSD post hoc was used to
analyze the diference between analyzed groups of Simmental
cows. Te diferences with p-values of p<0.05 were considered
signifcant (95% signifcance). Statistical analysis was carried
out by Ver.5.0 Microsoft Statistical Stat.Soft.Inc.1995.
Ethical compliance
Statement of ethical compliance: Te experiment was
done in compliance with Serbian Law on Animal Welfare
(Ofcial Gazette of the Republic of Serbia No 41/09) and
Ordinance on the conditions for registration for experimental
animals and the keeping of such register, training programs
on welfare on experimental animals, request forms for ap-
proval of conducting experiments on animals, standing,
treatment and killing experimental animals and reproduc-
tion, circulation, or implementation experiments on animals
(Ofcial Gazette of the Republic of Serbia No 39/10).
RESULTS
Results of the concentration of inorganic phosphorus in
serum of tested Simmental cows are shown in Table 2.
Signifcantly higher (p<0.05) levels of phosphorus were in
Table 2: Serum inorganic phosphorus concentrations (mmol/l) from
all the sampled cows (groups A, B and C)
Productive stage Group N X SD CV P<0.05
Close-up dry cows A 15 2.09 0.43 32.55
A:C
B:C
Fresh cows B 15 1.97 0.39 30.98
High-production cows C 15 2.40 0.38 18.93
Table 3: Serum magnesium concentration (mmol/l) from all the
sampled cows (groups A, B and C)
Productive stage Group N X SD CV P<0.05
Close-up dry cows A 15 1.02 0.26 43.25
A:C Fresh cows B 15 1.09 0.32 44.76
High-production cows C 15 1.28 0.20 29.23
Table 4: Serum calcium concentrations (mmol/l) from all the
sampled cows (groups A, B and C)
Productive stage Group N X SD CV P<0.05
Close-up dry cows A 15 2.35 0.24
21.98
B:C Fresh cows B 15 2.17 0.22
23.56
High-production cows C 15 2.53 0.26 19.23
N = Number of cows
X = Mean
SD = Standard deviation
CV = Coefcient of variance
Research Articles
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 Krsmanović, M. 14
group C compared to group B. Te mean serum phosphorus
concentration of close-up dry cows of group A was signif-
cantly higher (p<0.05) compared to fresh cows of group B.
Variations in serum inorganic phosphorus concentration was
not high within tested groups; the most pronounced vari-
ability was recorded in the fresh cows group (CV = 32.55%).
Results of serum magnesium concentration are shown
in Table 3.Te highest mean magnesium concentration
was found in high-production lactating Simmental cows
(1.28±0.20 mmol/l). Mean magnesium values were sig-
nifcantly higher (p<0.05) in high-production lactating
cows (group C) compared to close-up dry cows (group A).
Magnesium levels in the blood of tested cows had relatively
high variations within tested groups: in range from 0.20 to
0.32 mmol/l and the coefcient of variation varied from
29.30% to 44.76%.
Results of serum calcium concentrations are shown in
Table 4. Mean calcium concentration of fresh cows (group
B) was signifcantly lower in comparison with those of
high-production cows of group C (p<0.05). Te variability
of serum calcium concentrations among the three groups of
cows was not high, being most pronounced in the fresh cows
(CV=23.56%).
DISCUSSION
Lactating dairy cows use large quantities of calcium for
milk synthesis (11). Hypocalcemia in intensively managed
dairy cows causes periparturient paresis and contributes
to “downer cow syndrome”, while during lactation it can
also cause fertility disorders (2,6). Te results of the pres-
ent study indicate that in a group of Simmental dairy cows
in puerperium, lowest calcium levels were found, and these
values were signifcantly lower than the level of calcium in
the blood in Simmental cows in advanced pregnancy and
during maximum lactation. Tese fndings are similar to the
fndings of other researchers, who suggest that a sudden loss
of calcium from the body occurs in the puerperal cows, as for
every liter of colostrum 1.0-2.0 g of calcium is consumed (7),
while a physiological decline of calcium in the blood is held
steady for several days and on account of intensive produc-
tion, serum calcium decrease is correlated with the appear-
ance of puerperal paresis (7). Calcium metabolism is closely
related with phosphorus metabolism and is responsible for
the proper metabolism of vitamin D (12, 13).
Inorganic phosphorus levels in the blood of cows are at
low concentrations in the range of 2.1-3.0 mmol/l, while
signifcantly more phosphorus is bound to organic com-
pounds such as phospholipids (14, 15, 16). Similarly to serum
calcium, inorganic phosphorus values were signifcantly lower
in Simmental cows in puerperium compared to the values
of inorganic phosphorus in the blood of cows in advanced
pregnancy and during maximal lactation. Grünbergand co-
workers (17) found that hypophosphatemia is present in the
blood of highly productive Holstein-Friesian cows through-
out the postpartum period which is associated with the entry
of glucose in the glycolytic path of peripheral tissues, as well
as to supply the necessary quantities of mammary gland
phosphorus. Te authors believe that hypophosphatemia
allows proper glucose metabolism which prevents ketosis im-
mediately after calving (6, 18). Te sudden loss of phosphorus
from the body occurs in the puerperal cows when 0.8-1.9 g
phosphorus per liter of colostrum is spent. Tis physiological
decline of phosphorus in the blood is held steady for several
days and the intensity of phosphate fall is correlated with
previous fndings that phosphate blood level falls in older
animals (19).Te results of these tests show a decrease in
serum calcium and inorganic phosphorus in the blood of
Simmental cows in early lactation, suggesting increased use
of these macroelements by the mammary glands. Our results
show slightly higher calcium, phosphorus and magnesium
blood serum levels than in Jersey cows but similar to levels
of Holstein breed cows (12, 20), on the other hand calcium
and magnesium blood sera levels were found to be lower
than in beef cattle such as Angus, Charolais and Hereford,
or Limousin (10).
Magnesium, like calcium reduces neuro-muscular irrita-
bility and a drop in its concentration in the blood results in
spontaneous muscle contractions or tetany (1). Te results
of this study indicate that the lowest magnesium levels were
detected in Simmental cows in advanced pregnancy and that
greater values of magnesium in the blood were determined in
cows in puerperium. However the diferences of these values
were not signifcant. Te highest values of magnesium in the
blood were detected in high production Simmental cows
during the peak of lactation which were signifcantly higher
compared to the values of the cows in advanced pregnancy
and the puerperium. Te results obtained in this study in-
dicate that the magnesemia was within the normal range in
all tested cows, and lower values were found in a group of
cows in the periparturient period. Tis probably points to the
Research Articles
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 15 Macroelement Parameters of Simmental Cows
increased use of magnesium during negative energy balance
in early lactation.
CONCLUSION
Serum calcium and inorganic phosphorus in the blood of
Simmental dairy cows in puerperium were signifcantly
lower (p<0.05), compared to the values of cows in advanced
pregnancy and during maximum lactation, indicating the
increased use of these macro-elements by the mammary
gland of cows in early lactation. For the cows in the peri-
parturient period, values of magnesium in the blood were
signifcantly lower (p<0.05), compared to the values during
the periparturient period.
ACKNOWLEDGMENTS
Ministry of science, education and technological development of
the Republic of Serbia, projects number ID P 46002, TR 31071
and TR 31079.
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Research Articles