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Shabunin S.V., Vostroilova G.A., Khokhlova N.A., Shabanov D.I., Bliznetsova G.N., Ermakova T.I., Shaposhnikov I.T. Prospects for the application of extracellular proteinases of micromycete Aspergillus ochraceus in the treat-ment of mastitis in cows. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2022, Vol. 57, № 4, p. 706-717.
(how to cite this article)

doi: 10.15389/agrobiology.2022.4.706eng

UDC: 636.2.034:618.19-002:615.355:582.282.123.4

Acknowledgements:
Supported financially from the Russian Science Foundation (project No. 20-16-00085)

 

PROSPECTS FOR THE APPLICATION OF EXTRACELLULAR PROTEINASES OF MICROMYCETE Aspergillus ochraceus IN THE TREATMENT OF MASTITIS IN COWS

S.V. Shabunin, G.A. Vostroilova✉, N.A. Khokhlova, D.I. Shabanov,
G.N. Bliznetsova, T.I. Ermakova, I.T. Shaposhnikov

All-Russian Research Veterinary Institute of Pathology, Pharmacology and Therapy, 114-b, ul. Lomonosova, Voronezh, 394087 Russia, e-mail vnivipat@mail.ru, gvostroilova@mail.ru (✉ corresponding author), nina_xoxlova@mail.ru, am7d@mail.ru, gnbliznetsova@mail.ru, ermakova53@list.ru, 36011958@mail.ru

ORCID:
Shabunin S.V. orcid.org/0000-0002-2689-6998
Bliznetsova G.N. orcid.org/0000-0002-1042-9279
Vostroilova G.A. orcid.org/0000-0002-2960-038X
Ermakova T.I. orcid.org/0000-0003-1069-1223
Khokhlova N.A. orcid.org/0000-0001-6861-2554
Shaposhnikov I.T. orcid.org/0000-0003-0190-9083
Shabanov D.I. orcid.org/0000-0002-1574-1317

Received May 5, 2022

Cow mastitis, the etiological agents of which are pathogenic and opportunistic microorganisms, is considered one of the diseases that cause significant economic damage to dairy farming with a risk to the health of dairy consumers. With bacterial infections of the mammary gland in cows, the formation of protein exudates occurs. Proteinases that reduce the severity of the inflammatory response are included in the treatment regimens for various pathologies in medicine, but this practice is limited in veterinary medicine. In this work, we proved that a drug based on Aspergillus ochraceus proteinase increases the effectiveness of antibiotic therapy for cow mastitis. The aim of the study was to experimentally evaluate the possibility of using A. ochraceus BKM F-4104D micromycete proteinase in veterinary medicine. The studies were performed on lactating black-and-white cows (Bos taurus) with milk productivity for the previous lactation of 6900-7110 kg, which were divided into two groups (n = 16 each). All experimental animals were intracisternally injected with an anti-mastitis drug based on the beta-lactam antibiotic amoxicillin, clavulonic acid and prednisolone (ACP) at a dose of 3.0 g (one syringe dispenser) once per day for 3-4 days until the disappearance of clinical signs of mastitis. In the second group, 12 hours before the use of the ACP preparation, animals were additionally intracisternally administered a drug with the working name PAO-1. PAO-1 is an oil suspension containing 4.0 g (syringe-dosing device) extracellular proteinase of the micromycete A. ochraceus BKM F-4104D as an active ingredient. This proteinase is able to degrade heterogeneous protein substrates in a wide range of environmental conditions, which can increase the effectiveness of the etiotropic therapy of bovine mastitis. The condition of the breast, morpho-biochemical status were evaluated before treatment, after treatment and 7-10 days after the end of the administration of the drugs. It was found that the combined use of an antimicrobial agent and a preparation based on the proteinase of the micromycete A. ochraceus BKM F-4104D was accompanied by the recovery of 93.8 % of cows with clinical mastitis, which is 12.5 % higher (p < 0.05) than when using only the antimastitis drug ACP. Recovery of animals was characterized by normalization of morpho-biochemical status. The amount of β-globulins increased by 14.2 % (p < 0.05), triglycerides by 31.4 % (p < 0.05), creatinine decreased by 24.2 % (p < 0.05) compared to animals treated with ACP therapy. Endogenous intoxication and lipid peroxidation decreased, e.g., the concentration of malonic dialdehyde decreased by 40.4 % (p < 0.00005), medium-weight molecules by 46.3 % (p < 0.00005), NOx 3.6-fold ( p < 0.00005), endogenous intoxication index was 33.7 % lower (p < 0.005) compared to sick animals. The activity of the enzymatic and non-enzymatic components of the antioxidant defense increased, the concentration of vitamin A increased by 36.8 % (p < 0.005), vitamin E by 32.8 % (p < 0.05), vitamin C by 39.2 % (p < 0.005), catalase activity increased by 39.4 % (p < 0.005), glutathione peroxidase activity increased by 30.6 % (p < 0.005) compared to sick animals. Optimization of protein, lipid and mineral metabolism occurred. After the end of the therapeutic course, the number of somatic cells and their composition in the secret of the udder was normalized and there was no pathogenic microflora, which confirmes the complete clinical recovery of the cows. Our findings indicate that Aspergillus ochraceus BKM F-4104D micromycete proteinase which has high anticoagulant and fibrinolytic activity, can be very promising for the creation of domestically produced enzymatic veterinary drugs competitive in the world market.

Keywords: proteinases, Aspergillus ochraceus BKM F-4104D, PAO-1 drug, mastitis, cattle, enzyme preparations, combination therapy.

 

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