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Vol. 24 (2021 year), No. 3, DOI: 10.21443/1560-9278-2021-24-3
Kolberg N. A, Leontieva S. A., Tikhonov S. L., Tikhonova N. V., Shikhalev S. V., Kirpikova R. E.
Evaluation of the effect of enzymatic hydrolyzate of the bursa
of broiler chickens on cytotoxic activity and nonspecific immunity
The use of biologically active substances of tissue origin for the creation of biologically active additives and immunomodulatory drugs is an important area of scientific research in the field of nutrition and pharmacology. The aim of the work is to evaluate the effect of enzymatic hydrolysate from the fabricium bag of broiler chickens on the cytotoxicity and nonspecific immunity of mice against the background of experimental salmonella infection using biotechnological methods. For the experiment, three equal experimental groups of white mongrel mice were formed. All laboratory animals were fed daily for seven days with enzymatic hydrolysate at a dose of 750 mg/kg (therapeutic dose), 150 mg/kg (0.2 therapeutic dose) and 3,750 mg/kg (five therapeutic doses). At the same time, the control group of animals received intragastric water in the same volume. 24 hours after the last intake of the enzymatic hydrolysate, the animals were intraperitoneally infected with a culture of Salmonella enteritidis 92. The study reveals the absence of cytotoxic properties and impaired cell viability in cultures L929, J774.1A, HeLa S3, K562, and HST116 against the background of exposure to various concentrations of enzymatic hydrolysate from 0.02 to 10 mg/ml. It should be noted that there is a pronounced cytotoxic effect on MCF-7 tumor cells of the concentration of enzymatic hydrolysate in culture of 5 and 10 mg/ml, which suggests the possibility of using enzymatic hydrolysate for the prevention of tumor diseases. As a result of the experiment, it has been proved that the administration of enzymatic hydrolysate to mice increased the LD50 index and the survival rate of mice infected with the culture of the causative agent of salmonellosis by 1.5 times. Based on the conducted studies, it has been shown that the enzymatic hydrolysate of the lymphoid tissue of broiler chickens does not violate the viability of mammalian cells and does not exhibit cytotoxic properties on the metabolism of healthy mammalian cells, which indicates its safety. In an experiment on mice, the effect of enzymatic hydrolysate of the fabricium bag of broiler chickens on the activation of nonspecific immunity against salmonella infection has been revealed.
(in Russian, стр.7, fig. 0, tables. 3, ref 13, AdobePDF, AdobePDF 0 Kb)
Vol. 25 (2022 year), No. 3, DOI: 10.21443/1560-9278-2022-25-3
Kolberg N. A., Moskovenko N. V., Tikhonov S. L. , Tikhonova N. V.
The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens on the morphofunctional state of macrophages
The goal of studying the mechanisms of action of cells that structure the immune system is to obtain additional data about the influence of immunomodulating peptides of natural origin on the possibility of regulating immune processes. The effect of enzymatic hydrolysate of lymphoid tissue of broiler chickens (EHLTBC) on the viability of culture of peritoneal and alveolar macrophages, on the morphological characteristics and functional activity of macrophages has been studied. The effect of EHLTBC on the viability of culture of peritoneal and alveolar macrophages has been investigated. Morphometric characteristics of macrophages under the influence of EHLTBC have been given. The functional activity of peritoneal and alveolar macrophages has been assessed against the background of the introduction of EHLTBC. Enzymatic hydrolysate of lymphoid tissue of broiler chickens at concentrations of 1,5 and 10 % does not have a toxic effect on the culture of alveolar and peritoneal macrophages. EHLTBC enhances the phagocytic activity of peritoneal macrophages, which enhances their role in the chemotaxis and immune response. The stimulating effect of EHLTBC of poultry on the ability of macrophages to adhere has been revealed.
(in Russian, стр.14, fig. 9, tables. 0, ref 30, AdobePDF, AdobePDF 0 Kb)
Vol. 25 (2022 year), No. 3, DOI: 10.21443/1560-9278-2022-25-3
Tikhonov S. L., Tikhonova N. V., Kolberg N. A., Ozhgikhina A. S., Shikhalev S. V.
Microcapsulation and evaluation of subchronic toxicity of peptides extracted from cow colostrum and peptides of Fabricius bursa extract of broiler chickens
Peptides have high biological activity and are used in the composition of medicines. Peptides are not widely used in food technology as they are sensitive to the action of proteolytic enzymes of the gastrointestinal tract and are characterized by rapid decomposition reducing their effectiveness. Colostrum of cows is a promising source of peptides with different functional orientations. One of the ways contributing to the expansion of using peptides when administered orally is their microcapsulation. The aim of the research is to develop a technology and method for microcapsulating peptides extracted from cow colostrum after calving and to assess their subchronic toxicity. The development of an installation for microencapsulation of peptides was carried out taking into account the diameter of the wide part of the cone of the working chamber and the speed of soaring of small particles of peptide conglomerates. For the experiment on the comparative assessment of the subchronic toxicity of encapsulated and free peptides, 4 groups of nonlinear mice were formed, 10 in each. Group 1 of mice was a control, group 2 (experimental) received an extract of the fabricium bag of broiler chickens containing peptides of various molecular weights at a dose of 15,000 mg/kg, group 3 – free peptides extracted from cow colostrum serum in an amount of 1,500 mg/kg, group 4 – encapsulated peptides in an amount of 15,000 mg/kg. Hematological blood analysis was performed using the analyzer PCE-90Vet. The mice were weighed on the 7th, 9th, 14th, 21st and 28th days of the experiment. Biochemical analysis of animal blood sera was carried out using the analyzer StatFax 3300. During the study, it has been found that the treatment of peptides with maltodextrin solution for 1.0–1.5 minutes forms a protective coating with a thickness of more than 2 microns. The diameters of the wide and narrow parts of the cone of the device and the velocity of the gushing streams for the smallest and largest particles of the peptide conglomerate have been calculated. The absence of toxicity of microcapsulated peptides has been proven.
(in Russian, стр.11, fig. 3, tables. 3, ref 25, AdobePDF, AdobePDF 0 Kb)
Vol. 26 (2023 year), No. 3, DOI: 10.21443/1560-9278-2023-26-3
Merzlyakova N. V., Tikhonov S. L., Tikhonova N. V., Pestova I. G., Kulagova E. P.
Prediction of antimicrobial activity of a food peptide and evaluation of its effectiveness in vitro
Food antimicrobial peptides are increasingly relevant in the treatment of bacterial infections and have a number of advantages in comparison with drugs: slower emergence of resistance in bacterial strains, high activity against antibiotic films and immunomodulatory properties. The physico-chemical properties causing antimicrobial properties include: cationic charge, hydrophobicity, molecular weight and amino acid sequence. Promising sources of antimicrobial peptides are milk proteins, in particular, cow colostrum proteins. The aim of the work is to extract a peptide from the pepsin hydrolysate of cow colostrum, predict its antimicrobial activity using a bioinformatic approach and confirm its effectiveness in vitro. The molecular mass distribution of the peptide was evaluated by mass spectrometric method. The analysis of mass spectra was carried out using the Mascot program, the Peptide Fingerprint option – using the Protein NCBI database. Microsequencing was performed using a MiSeq sequencer. Modeling of the spatial structure of the isolated peptide was carried out using the Schrodinger Maestro molecular modeling program. The antimicrobial activity of the peptide was studied by the disco-diffusion method on gram-positive and gram-negative bacteria. The peptide has been isolated from the pepsin hydrolysate of cow colostrum, which is absent in the known proteomic bases Protein NCBI and AVPdb and, accordingly, its biological activity and functions have not been studied. The resulting peptide belongs to linear alpha-helical peptides consists of 11 amino acid sequences ANRKLRANKSR with a molecular weight of 8.2 kDa, an isoelectric point at 12.79, a charge of +5 (cationic), hydrophilicity (hydrophobicity) +20.84 Kcal*mol-1. As a result of in vitro studies, it was found that the isolated peptide has antimicrobial activity against E. coli ATCC 25922 and B. subtilis and antimicrobial action against C. albicans.
(in Russian, стр.10, fig. 3, tables. 2, ref 31, AdobePDF, AdobePDF 0 Kb)