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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

Protein content in milk of holstein black-and-white cows

N L Ignatieva* and E Yu Nemtseva

Department of general and private animal science, Chuvash State Agricultural
Academy, 29 K Marx Street, Cheboksary 428000, Chuvashia, Russian Federation
*
E-mail: [email protected]

Abstract. The most optimal method to solve the problem for cow's milk and protein content
increase is to carry out zootechnical and breeding activities. The goal was to study the milk
protein structure and content of the Holsteinized black-and-white breed cows of different
genotypes, as well as to establish the relationship nature between the activity of transamination
enzymes and the milk protein content of cows. For this purpose, 4 groups of experimental cows
(15 heads in each) were formed according to the principle of father's belonging to the countries
of origin (daughters of seed bulls of Canadian, Danish selection, Dutch and domestic
selection). The superiority of cows-daughters of foreign breeding bulls in terms of milk protein
content was established. Moreover, the milk of cows born from foreign producers contents
high level of casein - the most important fraction from the technological point of view.
Electrophoretic analysis of milk proteins isolated 16 fractions, including 9 casein and 7 whey
ones. The highest content was found in such fractions as αs1-, β-, αs2-, κ-caseins and β-Lg.
The calculated correlation coefficients between the alanine aminotransferase and aspartate
aminotransferase and the milk protein activity of cows showed a direct relationship between
them in cows of the studied groups. This is a favorable factor for increasing the cows milk
protein content.

1. Introduction
One of the most important sectors of the national economy remains agriculture. It is the main supplier
of food and raw materials for many industries, the basic source of income for the rural population. Its
normal functioning allows providing jobs to other sectors of the economy. In this regard, the effective
development of agriculture has important national, economic and social significance [1-3]. Estimates
of genetic parameters for milk yield in dairy cows are abundant in the literature [4-8]. There is a strong
belief that proteins are the most valuable constituents of milk. Their digestibility reaches 95%. Milk
proteins play a huge role for young growing organisms participating in the creation of new cells and
tissues as well as for mature people restoring obsolete cells [9]. Therefore, in all countries with
developed dairy cattle breeding, the assessment and selection according to protein content in milk is a
special direction in the practical selection of dairy cattle [10]. For the dairy industry, not only milk
protein yield but also milk protein composition is important; for example, milk with higher casein
content will result in greater cheese yield [11, 12]. Directional stock breeding to improve the structure
of milk protein is very relevant. These indicators affect the consumption of raw materials, time and
energy in the production of dairy products. The quality of products depends on them. It is well known
that the fractional composition of milk proteins is a genetic factor [13]. Thus, the increase of content
of technologically important protein fractions in milk and thereby improvement of technological
properties can only be done through targeted breeding. The main role in the rapid improvement of the
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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

productive qualities of black-and-white cattle is assigned to Holstein seed bulls. In this case the
assessment of seed bulls in a particular farm is one of the most productive and reliable elements in
breeding with a high-producing dairy herd.
Currently, the requirements for quality of dairy products and, in particular, for protein content have
increased. The dairy cattle breeding requires reliable and accurate indicators of its forecast. It is known
that enzymatic blood systems under conditions of intensification of metabolic processes in animals
play an important role. They are directly related to the course of milk formation. Therefore, the
application of the interior characteristics of an animal as an innovative breeding method of modern
biochemical genetics is quite promising. Learning the proportion of interdependence of indicators of
protein metabolism and milk productivity in dairy cattle is relevant. It is an opportunity to raise milk
productivity and to increase the content of fat and protein in cow’s milk.
Such blood enzymes as alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT)
can be used in breeding with dairy cattle [14]. The purpose of the research is to study the content and
structure of proteins in the milk of Holstein black-and-white cows of different genotypes as well as to
establish the nature of relationship between the activity of transamination enzymes and the protein
content in cow’s milk.

2. Materials and methods

The Federal State Unitary Enterprise Scientific-Experimental Farm “Privolzhskoye” of the Chuvash
State Agricultural Academy where the studies have been conducted ranches Holstein black-and-white
cows. The herd is represented by the daughters of Holstein bulls from different countries of origin.
Four groups of experimental cows (15 animals each) were composed according to the principle of
father’s belonging to countries of origin. The experimental groups of cows were made up by the
method of pair-analogues taking into account blood relationship, live weight, age and physiological
state. The daughters of Canadian seed bulls formed the first group, Danish – the second one. Their
herdmates of Dutch and domestic selection were included into the third and fourth groups,
respectively. The primary research materials have become cows breeding cards, automated system of
livestock zootechnical registration on PEVM ARM "SELEX" and other documents of zootechnical
registration. The experimental animals were in the same conditions of feeding and keeping.
Analytical studies were conducted in the laboratory of the Department of Private Zootechnics of
the Chuvash State Agricultural Academy using modern instruments and equipment. The total protein
content was determined in an average daily milk sample taken in proportion to the yield of milk (by
the method of formol titration). Milk proteins were fractioned by electrophoretic method in a synthetic
polyacrylamide gel which makes it possible to carry out qualitative and quantitative analysis of milk
proteins faster and more qualitatively in the modern mini-chamber for vertical electrophoresis with
glass size of 2020 VE-20 (Helicon) [15].
The gels were examined using the high-frequency Vilber Lourmat TFP-L/WL transilluminator
(Helicon). Quantitative analysis of phoregrams of the studied samples was carried out by scanning
using the computer video system Gel Imager-2, designed for capturing and processing images of
luminescent DNA in gels (Helicon).
Blood for research was taken from the jugular vein located in the jugular gutter. Pre-sterilized
special injection needles were used for the collection. The activity of the transamination enzymes
ASAT and ALAT in non-homolized blood serum was determined photometrically with a unified
Wright-Frenkel method using set of reagents AlAT-Vital and AsAT-Vital.

3. Results and discussion

The studies have shown that the milk of cows from different experimental groups had some
differences. Animals of group II (3.24%) were characterized by the highest protein content in milk. It
was significantly higher than in animals of group IV by 0.24% (P < 0.05). The protein content in milk
of cows of experimental groups I and III made up 3.15 and 3.16%, respectively. The highest casein
content (2.52%) was found in the milk of cows of experimental group II. Moreover, the descendants of

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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

Canadian and Dutch bulls (group I and III) were characterized by an equal casein content of 2.46%. It
is 0.12% more than cows of group IV. However, the difference is not reliable. No significant
differences in the level of whey protein in milk of cows of different origin were found. The Danish
cows (group II) showed 0.72% of whey protein content in milk which was 0.06% higher than in
domestic cows (group IV) (P <0.05). It is 0.04% higher in Dutch cows (group III), and 0.03% higher
in daughters of Canadian bulls (group I) than in animals of group IV (the offspring of domestic
selection bulls). The established superiority of cows of imported bulls in terms of protein content in
milk allows us to recommend using them as improvers of protein content in milk of Holstein black-
and-white cows.
The studies of electrophoretic mobility have identified individual protein fractions of milk: 9 casein
and 7 serum. αs1-, β-, αs2- and κ-caseins identified on the electrophoregram of casein and β-Lg - on the
electrophoregram of whey proteins had the highest concentration. The share of other fractions was
insignificant.
It is determined that the electrophoretic presentation of milk proteins separated on the carrier
material - polyacrylamide gel - in cows of different origin is different. According to the current
classification a detailed analysis of milk protein fractions is shown in figure 1.

Figure 1. The structure of milk protein in Holstein black-and-white cows.

It can be seen that cows of group III show significant differences in the concentration of the most
important protein fraction of casein (79.53% of the total protein). Whereas the domestic origin
herdmates (group IV) have a casein concentration of 78.57%. The concentration of such casein
fractions as αs1-, β- and κ-fractions form the quality milk products containing protein. Milk of the
domestic origin cows (experimental group IV) in comparison with milk from other groups is
characterized by a lower concentration of αs1-casein (10.67%).
By αs1-casein content, the daughters of Canadian, Danish and Dutch bulls exceeded domestic
breeding cows (group IV) by 0.4, 0.13 and 0.9%, respectively. The domestic origin cows also had a
low β-casein content of 12.03%. The difference in the content of this fraction in the descendants of
foreign (I, II and III group) and domestic (IV group) bulls made up 0.47, 0.84 and 1.0%, respectively.
The concentration of κ-casein at the level of 8.63 and 8.83% was determined in the milk of cows in the

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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

II and III experimental groups (Danish and Dutch breeding). Such a milk protein fraction as γ-casein is
considered to be unimportant from a technological point of view. Its share in the structure of milk
protein was greater in daughters of the domestic (group IV) bulls than in other groups of animals.
Whereas the concentration of γ-casein was insignificant in cows of Dutch origin (experimental group
III), at the level of 7.50% of the total milk protein content.
Immunoglobulins are a fraction of whey proteins. The results of the analysis showed that their
relative content in milk of daughters of the domestic bulls (group IV) was the highest (2.77%). The
lowest concentration was noted in milk of Dutch cows (group III) - at the level of 2.43%. The β-Lg
and α-La fractions are identified between the fast fraction (F) and serum albumin. The concentration
of β-Lg is at the level of 3.67% in Canadian cows which is 0.23, 0.4 and 0.16% less than in groups II,
III and IV, respectively. The smallest concentration of α-La was also found in cows of experimental
group I - daughters of Canadian bulls.
Currently, the issue of selection of dairy cattle to increase the content of high-value milk protein is
relevant. Therefore, the identification of the interdependence of the content of protein fractions will
solve this difficult problem [1].
It is discovered that protein content in milk of all the studied groups of animals is closely related to
the level of αs2-casein (the correlation coefficient was in the range from 0.50 to 0.97). This creates
suitable conditions for practical selection. A negative relationship between the relative content of total
protein and γ casein in the studied groups of cows (the correlation coefficient ranged from -0.28 to -
0.99) indicates an opposite relationship. The higher the total protein value is, the lower the γ value of
the casein fraction and vice versa. The relationship of total protein and other protein fractions in these
studies is not found.
The absolute values of the correlation coefficients of the total protein and the relative content of
complex proteins in animals of groups I and IV make up 0.57 and 0.68 for casein, -0.57 and -0.68 for
serum proteins. It is interpreted as follows: the selection of dairy cattle by content of protein in milk in
the group of Canadian and domestic animals will lead to an increase in the proportion of casein in the
total protein. In this case, the concentration of whey proteins will decrease. This connection is
desirable for cheese production technology as it will lead to an increase in the yield of cheese mass.
The data of the correlation analysis of the dependence of the total protein and the relative content of
complex proteins in animals of groups II and III turned out to be opposite to what was observed in
animals of groups I and IV. The correlation of the total protein and the main fractions of casein (αs1-,
β- and κ) is positive in the daughters of Canadian and domestic fathers (I and IV groups). The
interdependence of total protein and casein β-fraction was also established in Danish cows - group II
(with the increase of one, the other criteria also increases). Moreover, the relationship between the
total protein content and αs1 and κ fraction of casein in this group of cows is inverse (an increase in one
criteria leads to a decrease in the other). The correlation analysis revealed a positive relationship
between the concentration of total protein and αs1-fraction and a negative relationship with its β- and
κ-fractions in daughters of Dutch bulls (group III).
The calculation of the coefficients of dependence (correlation) between the total protein content
and the main fractions of whey proteins – β-Lg and α-La – showed the presence of a positive
relationship in cows of groups I, II and IV. The values of the correlation coefficients ranged from 0.33
to 0.95. Moreover, experimental group of animals No. 3 was characterized by a weak (–0.38) and
average (–0.73) dependence of the total protein content and the relative content of β-Lg and α-La.
Moreover, the reliability of the correlation is absent.
The breeding work can also use such transaminase enzymes as alanine aminotransferase and
aspartate aminotransferase. The biochemical profile of enzymes also depends on the stage of lactation
of animals. The change in the activity of the enzymes ALAT and ASAT during lactation is presented
in figures 2 and 3.

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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

Figure 2. Change of ALAT activity in serum during lactation.

The analysis shows that the catalytic activity of the enzyme ALAT in the first three months of
lactation in cows of all experimental groups increases. Then, at the 4th month of lactation, a sharp
decline in enzyme activity is observed. From the fifth month of lactation, daughters of foreign bulls
(groups I, II and III) have a significant decrease in its activity with some fluctuations (fluctuations are
unreliable). It should be noted that animals of group IV (daughters of domestic bulls) are characterized
by a decrease in enzyme activity up to the 7th month of lactation.

Figure 3. Change of AsAT activity in serum during lactation.

According to the change in the activity of ASAT enzyme during lactation in cows of different
origin, the following was noted: increases until the 6th month of lactation, then decreases by the end of
lactation.

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International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

Since the biochemical composition of the blood is to some extent interconnected with the breeding
and productive qualities of animals, the correlation coefficients of two values - the activity of ALAT
and ASAT enzymes and the protein content in milk - were calculated (table 1).
Table 1. The relationship of ALAT and ASAT with protein content in milk of
cows (Х±Sx).
Enzyme Activity, mcM/(h*ml)
Animal Groups ALAT ASAT
I 0.98±0.22*** 0.99±0.13***
II 0.78±0.44 0.97±0.16***
III 0.92±0.39* 0.96±0.26***
IV 0.27±0.56 0.37±0.54
*Р <0.05
***Р <0.001
The studies conducted on Holstein black-and-white cows of different origin showed that a positive
statistically significant correlation was established between the activity of ASAT and ALAT in the
blood and the protein level in cow’s milk. The maximum value of correlation was found in the group
of cows originating from bulls of Canadian selection. The second in correlation are cows of group III -
of Dutch origin. It is a favorable factor for increasing protein content in milk of cows.

4. Conclusion
As a result of the studies, the superiority of cows-daughters of the imported bulls- in terms of the
protein content in milk was established. It allows us to recommend using them as improvers of protein
content in milk of Holstein black-and-white cows. The detailed analysis of the fractional composition
of the milk protein of cows of different origin indicates that the female ancestors of Holstein producers
of Dutch companies are distinguished by the highest content of the main protein fractions that affect
the technology of milk and dairy products. The protein content in milk of cows positively correlates
with the enzymatic activity of serum transaminases. It is a favorable factor for increasing protein
content in milk of cows.

References
[1] Lavrentyev A Y, Nikolaeva A I, Evdokimov N V, Semenov V G, Sherne V S, Nemtseva E Y,
Danilova N V, Petrov N S and Toboyev G M 2019 The efficiency of the use of “Biostrong
510” additive in the technology of broiler chicken meat production. IOP Conference Series:
Earth and Environmental Science Macau 346 012053 DOI: 10.1088/1755-
1315/346/1/012053
[2] Sherne V S, Lavrentev A Y, Evdokimov N V, Petrov N S, Nemtseva E Y and Danilova N V
2019 Enzyme preparations in compound feed for ducklings. IOP Conference Series: Earth
and Environmental Science 346 012030 DOI: 10.1088/1755-1315/346/1/012030
[3] Evdokimov N V, Lavrentyev A Y, Alekseev V A, Sherne V S, Nemtseva E Y, Petrov N S,
Danilova N V, Novikov A A and Toboev G M 2019 The possibility of using immunogenetic
criteria for the characterization of the breed, to predict the productive qualities and results of
the selection of pigs of the tsivilsky breed. IOP Conference Series: Earth and Environmental
Science 346 012052 DOI: 10.1088/1755-1315/346/1/012052
[4] Hammami H, Rekik B, Soyeurt H, Bastin C, Stoll J and Gengler N 2008 Genotype Environment
Interaction for Milk yield in Holsteins Using Luxembourg and Tunisian Populations. J.
Dairy Sci. 91(9) 3661 DOI: 10.3168/jds.2008-1147
[5] Hammami H, Rekik B, Bastin C, Soyeurt H, Bormann J, Stoll J and Gengler N 2009
Environmental Sensitivity for MilkYield in Luxembourg and Tunisian Holsteins by Herd
Management Level. J. Dairy Sci. 92(9) 4604 DOI: 10.3168/jds.2008-1513

6
International AgroScience Conference (AgroScience-2020) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 604 (2020) 012025 doi:10.1088/1755-1315/604/1/012025

[6] Ben Gara A, Rekik B and Bouallègue M 2006 Genetic parameters and evaluation of Tunisian
dairy cattle population for milk yield by Bayesian and BLUP analyses. Liv. Sci. 100 142
DOI: 10.1016/j.livprodsci.2005.08.012
[7] Ben Gara A, Jemmali B, Hammami H, Rouissi H, Bouallegue M and Rekik B 2012 Milk
Production of Holsteins Under Mediterranean Conditions: Case of the Tunisian Population
Milk production (NY, USA) p 1-12
[8] Ilahi H, Hammouda M B and Othmane M 2012 Bayesian genetic analysis of milk yield in
Tunisian Holstein dairy cattle population. Open Journal of Genetics 2(02) 103
DOI: 10.4236/ojgen.2012.22015
[9] Madureira A R, Pereira C I, Gomes A M P, Pintado M E and Malcata X 2007 Bovine whey
proteins-оverview on the main biological properties. Food Res Int. 40 1197
DOI: 10.1016/j.foodres.2007.07.005
[10] Ignatieva N L, Aizatov R M 2012 Protein content in milk of cows of different genetic origin.
Scientific notes of the Kazan State Academy of Veterinary Medicine named after
N E Bauman 209 128
[11] Emmons D B, Ernstrom C A, Lacroix C and Verret P 1990 Predictive formulae for yield of
cheese from composition of milk. J. Dairy Sci. 73 1365 DOI: 10.3168/jds.S0022-
0302(90)78803-0
[12] Wedholm A, Larsen L B, Lindmark-Månsson H, Karlsson A H and Andrén A 2006 Effect of
protein composition on the cheesemaking properties of milk from individual dairy cows
J. Dairy Sci. 89(9) 3296 DOI: 10.3168/jds.S0022-0302(06)72366-9
[13] Martin P, Szymanowska M, Zwierzchowski L and Leroux C 2002 The impact of genetic
polymorphisms on the protein composition of ruminant milks. Reprod Nutr. Dev. 42 433
DOI: 10.1051/rnd:2002036
[14] Ignatieva N L and Aizatov R M 2011 Protein-milk Content of cows of different origin and its
relation to the activity of transaminase enzymes. Bulletin of the Ulyanovsk state agricultural
Academy 3(15) 81
[15] Khayertdinov R A, Afanasyev M P and Khayertdinov R R2009 Proteins of milk (Kazan:
Publishing house Idel-Press) p. 256