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Vertiprakhov V.G., Selionova M.I., Malorodov V.V., Laptev G.Yu., Ilyina L.A. Milk trypsin clear increases under bovine mastitis simultaneously with inflammation gene expression. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2023, Vol. 58, № 4, p. 685-699.
(how to cite this article)

doi: 10.15389/agrobiology.2023.4.685eng

UDC: 636.2.034:618.19-002:591.1:577.2

 

MILK TRYPSIN CLEAR INCREASES UNDER BOVINE MASTITIS SIMULTANEOUSLY WITH INFLAMMATION GENE EXPRESSION

V.G. Vertiprakhov1 ✉, M.I. Selionova1, V.V. Malorodov1, G.Yu. Laptev2, L.A. Ilyina2

1Russian State Agrarian University — Timiryazev Moscow Agricultural Academy, 49, ul. Timiryazevskaya, Moscow, 127550 Russia, e-mail Vertiprakhov63@mail.ru (✉ corresponding author), m_selin@mail.ru, malorodov56@gmail.com;
2Saint-Petersburg State Agrarian University, 2, Peterburgskoe sh., St. Petersburg—Pushkin, 196601 Russia, e-mail georg-laptev@rambler.ru, ilina@biotrof.ru

ORCID:
BVertiprakhov V.G. orcid.org/0000-0002-3240-7636
Laptev G.Yu. orcid.org/0000-0002-8795-6659
Selionova M.I. orcid.org/0000-0002-9501-8080
Ilyina L.A. orcid.org/0000-0003-2789-4844
Malorodov V.V. orcid.org/0000-0001-9033-7552

Final revision received March 30, 2023
Accepted June 9, 2023

Mastitis is one of the most serious problems in dairy farming. Mastitis often affects high-yielding cows, with a 10-15 % reduction in productivity and irreversible mammary gland dysfunction. In clinical course pathology has clear diagnostic signs. The main known methods of diagnostics of subclinical forms of mastitis (mastitis tests) are based on the determination of somatic cells in milk, the number of which correlates with inflammation, but the development of methods for early diagnosis of mastitis and pre-mastitis state of cows remains relevant. Biochemical parameters and morphological profiles of animal blood, expression of genes associated with inflammation are also examined in mastitis. However, the presence of enzymes in animal milk has not been fully studied. Trypsin is considered as a hormone-like substance capable of influencing metabolism and being a marker of inflammatory processes in animals and humans. Previously, we have shown the role of trypsin in experimental toxicosis of chickens and dietary changes. In the presented study we have for the first time revealed trypsin in the milk of cows, the increase in its activity in mastitis was established and compared with changes in other indicators used to assess the state of animals in pathology. The aim of the present work is to detect trypsin activity in milk of healthy and mastitis-affected cows and to determine the number of somatic cells in milk, relative expression of genes associated with inflammation, as well as morpho-biochemical blood parameters. The results obtained on Ayrshire cows (Bos taurus), 10 lactating cows without clinical signs of mastitis and 15 cows with clinical signs of mastitis (SGC Smena — a branch of the FSC VNITIP RAS, Moscow Province, 2022), showed that in the milk, the activity of genes associated with inflammation and the trypsin activity varied depending on the mammary gland health. In mastitis this index increased compared to the norm by 106.6 % (p £ 0.05), whereas trypsin activity in blood serum of healthy and mastitis cows had no significant differences. Of the biochemical parameters of cow blood, the most informative were the concentration of glucose, calcium and phosphorus. We found that in blood serum of mastitic cows the amount of glucose increases by 67.4% (p < 0.05), calcium by 38.8% (p < 0.05), the concentration of phosphorus, on the contrary, decreases by 23.8 % (p < 0.05) compared to healthy animals. In the blood morphological profile at mastitis leukocytosis is observed, there is a decrease in immunoreactivity by 42,5 % (p < 0.05), the ratio of lymphocytes and neutrophils by 20.4 % (p < 0.05), the number of eosinophils by 57.4 % (p < 0.05) and basophils by 33.3 % (p < 0.05), while the number of monocytes increases by 46.5 % compared to the control (p < 0.05). The expression of genes of monocyte chemotactic protein 1 and monocyte chemotactic protein 2 increased 5.5-fold, tumor necrosis factor alpha 3.9-fold, interleukin 4 and interleukin 8 2.9-fold and 14-fold, respectively, in cows with mastitis compared to healthy cows. Thus, we found that cow's milk contains trypsin, which is not inferior to the enzyme in the blood serum of animals in terms of activity (48.2±3.8 units/l). In inflammation of the mammary gland confirmed by instrumental and molecular genetic methods, trypsin activity in milk increases, which can be used in the development of diagnostic methods for pre-mastitis and early stages of mastitis.

Keywords: cows, mastitis, milk trypsin, mastitis diagnostic methods.

 

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