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Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv
original article I UDC 619.22.28: 614.48: 615.9: 636.065 | doi: 10.32718/ujvas5-1.08
Volume 5 Number 1
The activity of T- and B-cell links of specific protection of chicken-broilers under the influence of synbiotic preparation "Biomagn" and "Diolide" disinfectant
O. M. Chechet1 , V. L. Kovalenko2 ,0.1. Vishchur3 , O. S. Haidei1 , N. V. Liniichuk1 , B.V. Gutyj4 ■■:,(). V. Krushelnytska4 ..:
1State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise, Donetska Str., 30, Kyiv, 03151, Ukraine
2State Scientific Control Institute Biotechnology and Strains of Microorganisms, Donetska Str., 30, Kyiv, 03151, Ukraine 3Institute of Animal Biology NAAS, V. Stusa Str., 38, Lviv, 79034, Ukraine
4Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Pekarska Str., 50, Lviv, 79010, Ukraine
Article info Received 16.02.2022 Received in revised form
17.03.2022 Accepted 18.03.2022
Correspondence author Olha Haidei
Tel.: +38-067-171-15-58 E-mail: olga.gaidei@gmail. com
2022 Chechet O. et al. This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
One of the most actual problems of modern poultry farming is to increase the preservation of chickens and ensure high intensity of their growth at all stages of rearing. The development of alternative methods of prevention and treatment of infectious diseases of poultry involves the use of preparations of natural or organic origin, which have antagonistic properties against infectious agents and are able to balance the immune response. A prominent place in the system of disease prevention and increasing the resistance of animals and poultry is the development of modern probiotic and bactericidal preparations. In view of this, we studied the influence of the synbiotic preparations "Biomagn" in combination with the disinfectant "Diolide" on the state of the T- and B-cell links of the specific immunity of broiler chickens during the period of their rearing. The preparation "Biomagn" is based on representatives of the normal commensal microflora - non-pathogenic lactic acid bacteria with antibacterial and immunomodulatory properties and additional components: magnesium chloride, chitosan, xylanase, protease, cellulase, milk thistle meal, acidity regulator, betaine and emulsifier. The specified remedy is used to ensure optimal level of metabolism, increase immune function, growth, safety of animals and poultry. The results of the research showed that the use of the synbiotic preparation "Biomagn" in combination with the disinfectant "Di-olide" had a positive influence on the activity of T- and B-cell links of specific defense of broiler chickens organism during the period of their rearing. This is evidenced by an increase in the amount of T-lymphocytes (total, active and theophylline-resistant) and B-lymphocytes in the blood and an increase in their functional activity due to the redistribution of the receptor apparatus of immunocompetent cells. In particular, a decrease in the number of functionally inactive T- and B-lymphocytes in the blood and an increase in the number of cells with low and medium degree of avidity. Therefore, the use of a set of studied immunotropic preparations is a promising direction in increasing the resistance and productivity of poultry. This allows in industrial conditions of maintenance, without loss of productivity, to respond to technological factors that can lead to immunodeficiency, disease and death of poultry.
Keywords: broiler chickens, pro- and prebiotics, chlorine dioxide, immunity, nonspecific resistance._
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Citation:
Chechet, O. M., Kovalenko, V. L., Vishchur, O. I., Haidei, O. S., Liniichuk, N. V., Gutyj, B. V., & Krushelnytska, O. V. (2022). The activity of T- and B-cell links of specific protection of chicken-broilers under the influence of synbiotic preparation "Biomagn" and "Diolide" disinfectant. Ukrainian Journal of Veterinary and Agricultural Sciences, 5(1), 46-52.
Contents
1. Introduction................. .. 46
2. Materials and methods .... .. 47
3. Results and discussion .... .. 48
4. Conclusions................ .. 51
.. 51
1. Introduction
It is known that low resistance of chickens at an early
age is due to their biological characteristics, high concentration of livestock, the negative influence of technological factors and poorly balanced feeding, which leads to oxida-tive stress, immunodeficiency and death of poultry (Sobolev et al., 2020; 2021; Brezvyn et al., 2021).
At the same time pathogens of pathogenic microflora are also one of the causes of disease in poultry, especially in the
gastrointestinal tract. Widespread use of antibacterial preparations leads to the artificial formation of antibiotic resistance in the microflora and the difficulty of treating infectious diseases, violation of microbiocinosis, which has a negative influence on the productivity of poultry (Kalinichenko, 2013). Antibiotics, such as chloramphenicol, tetracycline, aminoglycosides, which are used for poultry in therapeutic doses, has a negative influence on the formation of immunity. These preparations suppress the normal intestinal microflora, which forms various biologically active
substances and regulates the body's immune function. Pathogens of viruses and bacteria also reduce immunobiological reactivity, which leads to disease.
In recent years, states of European Union, as well as the United States, have tightened controls and significantly reduced the use of feed antibiotics in livestock, and a number of preparations have been banned to prevent them from entering food. Currently, feed antibiotics are added only in the production of premixes for medicinal purposes and pre-starter feed for poultry and pigs. Therefore, synbiotic supplements may be an important alternative to feed antibiotics (Barratt & Rodford, 2001; Frei et al., 2015; Manzanares et al., 2016; Sarao & Arora, 2017).
The development of preparations based on normal microflora, in particular strains of lacto- and bifidobacteria (Mokrozub et al., 2012) and their use has a positive influence on the body's immune function, in particular, contributes to the increase of the phagocytic activity of monocytes, neutrophils and others (Tizard, 2009). Thus, it is necessary to develop and implement alternative methods of prevention and treatment of infectious diseases with preparations of natural origin with antagonistic properties against infectious agents. Modern probiotics in combination with safe bactericidal drugs will enhance antibacterial and immunomodulatory properties to protect the poultry's body (Kalinichenko, 2013).
It is known that a promising direction of correction of dysbacteriosis are compositions based on probiotic strains of bacilli where strains Bacillus subtilis, Bacillus licheniformis, Enterococcus faecium are most commonly used and dried fermentation products of Lactococcus Lactis, Bacillus sub-tilis, Bacillus and other substances that form a synbiotic. Synbiotic is a combined preparation in which probiotics and prebiotics have a mutually reinforcing effect on physiological functions and metabolic processes in the body of poultry. Strains as a part of probiotic preparations produce large amounts of antibiotics and other substances that suppress many microorganisms. Bacteria of the genus Bacillus show a variety of antimicrobial activity and in addition to therapeutic and prophylactic properties, often have enzymatic properties. Therefore, their use improves feed digestibility (Stoianovskyi & Kolomiiets, 2011; Mokrozub et al., 2012; Kurtiak & Romanovych, 2015). At the same time, the long-term effects of the immune system's response to random microorganisms, such as spore-forming bacteria, remain unknown. Further research is needed to clarify the mechanisms of their probiotic action on living systems.
In the complex of veterinary and sanitary measures in poultry farming, disinfection occupies an important place in order to prevent and eliminate dangerous infectious diseases of poultry. At the same time, it is important to clean and disinfect the drinking water supply system, as microorganisms can precipitate together with vitamins and other additives (Ma et al., 2012; Tsynovyi, 2013). One of the effective components of safe substances is chlorine dioxide. Chlorine dioxide in doses from 1.0 to 1.5 mg/dm3 is an effective mean for a wide range of viruses and bacteria, respectively, for disinfection of drinking water depending on the level of contamination (Sorlini et al., 2014).
Of course, in the observance of sanitation it is necessary to use highly effective environmentally friendly preparations that have a prolonged bactericidal action. Therefore, it is important for poultry to develop and implement new disinfectants that are easy to use, non-toxic, non-carcinogenic, with a broad bactericidal spectrum, do not cause addiction to
the microflora and provide a permanent bactericidal action (Sanekata et al., 2010; Yu et al., 2014; Sorlini et al., 2014).
The immune system is a complex of protection of organs and tissues, which with other body systems maintain a specific homeostasis of the internal environment (Tizard, 2009; Mokrozub et al., 2012; Kalinichenko, 2013).
Natural immunity is inherent in many living organisms, including plants, insects and even bacteria, which have protective mechanisms against bacteriophage viruses. Specific (adaptive) immunity is inherent only in higher animals and reaches the most complex organization in warm-blooded animals (birds and mammals). Natural immunity acts quickly because it does not depend on clonal proliferation of antigen-specific cells inherent in adaptive immunity, and is usually effective. Natural immunity cells play an important role in adaptive immunity - some of them (dendritic cells) are antigen-presenting cells capable of stimulating antigen-specific cells of adaptive immunity (T- and B-lymphocytes) (Romanyuk & Komisarenko, 2012).
From the data of the literature it is known about the influence of various immunotropic preparations on T- and B-cell immunity (Mudrak & Vishchur, 2011; Hrabovskyi & Hrabovska, 2015), the mechanism of development of reactions of homeostatic systems to external stimuli (Kaminska et al., 2008) and, in particular, in poultry (Kuzmenko, 2011; Marvar et al., 2012).
However, it is necessary to clarify the mechanisms of influence of new synbiotic preparations and disinfectants to activate the cellular component of specific protection of the organism and to develop schemes for their use in the technological system of industrial poultry farming. This is due to the relevance of expanding and deepening research aimed at studying the age characteristics of the formation of immun-obiological reactivity in the organism of poultry, which underlie its growth and development, mechanisms and factors of their regulation.
The purpose of the work was to determine the influence of the synbiotic "Biomagn" in combination with an aqueous solution of the disinfectant "Diolide" on the activity of T- and B-cell units of specific immunity of broiler chickens during the period of their rearing.
2. Materials and methods
The investigation was conducted in one of the farms of the Lviv region on broiler chickens cross R0SS-308, from 1 to 41 days of age. Chickens were kept in poultry houses with free access to feed and water, technological parameters of broiler breeding (temperature and light regime) in accordance with the norms of 0NTP-2005. 2 groups of broiler chickens were formed for research: control and experimental groups, 100 heads in each. The broilers of the control group were fed standard compound feed (SCF) according to the existing norms recommended for the R0SS-308 cross. The chickens of the experimental group were similarly fed SC and synbiotic preparation "Biomagn" at the rate of 0.5 kg per ton of combined feed. This preparation was used according to the following scheme: the first time at one day of age - seven days in a row, the next task - at 22 days of age, seven days in a row).
The preparation "Biomagn" is a mixture of probiotic bacteria: Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans, Enterococcus faecium and dried fermentation products of microorganisms: Lactococcus Lactis, Bacillus
subtilis, Bacillus milk thistle, acidity regulator, betaine and emulsifier (TU U 24.2-00699690-003: 2022).
At the same time, the broilers of the experimental group throughout the experiment were fed with water a solution of the preparation "Diolide" (TU U 24.2-00699690-001: 2022), where the main active ingredients: sodium chlorite, sodium chloride. For disinfection of drinking water, the disinfectant was used at a dose of 1.0 mg/L of chlorine dioxide. "Diolide" is added through the mediators, pre-uterine solution is diluted to a concentration that allows it to be presented by the medicator in accordance with its technical characteristics. This preparation was developed by the staff of the State Research Institute for Laboratory Diagnostics and Veterinary Sanitary Examination (Kyiv, SRILDVSE).
For immunological investigations, chickens were bled at different ages: 16-, 27-, 34- and 41-day-old.
The relative numbers of T and B lymphocytes and their individual populations in the reaction of spontaneous rosette formation with ram erythrocytes as markers were determined in the blood of broiler chickens. The principle of the method is the ability of lymphocytes to form so-called "sockets" with heterogeneous erythrocytes. In the center of the "socket" is a lymphocyte, the membrane of which contains specific receptors, and on the periphery of erythrocytes. The degree of activity of lymphocytes is judged by the number of erythrocytes adsorbed by one lymphocyte. The relative number of total (TE-RUL - total rosette-forming lymphocytes) and active T-lymphocytes (TA-RUL - active rosette-forming lymphocytes) was determined. Broiler chicken lymphocytes were isolated from heparin-stabilized blood in a ficol-verografin gradient with a relative density of 1.077. T-lymphocytes were determined in the reaction of spontaneous rosette formation with ram erythrocytes by M. Jondal et al., 1972 in a modification (Vlizlo, 2012). Active rosette-forming lymphocytes with receptors capable of attaching erythrocytes without incubation and theophylline-resistant (TLR) lymphocytes that form rosettes after incubation with theophylline were isolated. The smears were dried, fixed with methanol, and stained for 7-10 min according to Romanowski-Gimza. Smear microscopy was performed under immersion at a magnification of 90 H 7. Lymphocytes were distinguished by the number of attached erythrocytes: zero - did not attach any, poorly differentiated (low-avid lymphocytes or cells with low surface receptor density) - 3-5 erythrocytes were joined, average avid subpopulations - 6-10 erythrocytes, highly differentiated (highly avid) - "rosettes" with more than 10 erythrocytes (M - morula) (Vlizlo, 2012).
The determination of the relative number of theophylline-resistant T-helpers is based on the fact that these cells
carry on their surface receptors for class M immunoglobulins and T-suppressors for class G immunoglobulins. The number of theophylline-sensitive T-suppressor lymphocytes was determined by the difference between the number of TE-RUL and T-helpers.
The relative number of B-lymphocytes was determined, the method of identification of which is based on the presence of membrane immunoglobulin receptors in them, which ensures the attachment of indicator cells to B-lymphocytes, which contain complement-antigen-complex (EAS-RUL). Ram erythrocytes sensitized with antibodies and complement were used as indicator cells. Ready liquid hemolytic serum (titer 1: 1 200) and ready dry guinea pig complement were used to prepare the complement antigen complex.
All investigations were performed according to the methods described in the guide (Starovoitova et al., 2012).
The experimental part of the work was carried out taking into account the "General Ethical Principles of Animal Experiments", approved by the National Congress on Bioethics (Kyiv, 2001) and agreed with the provisions of the "European Convention for the Protection of Vertebrate Animals" which are used for experimental and other scientific purposes (Strasbourg, 1985).
Digital data were processed by biometric method of var-iational nonparametric analysis using Microsoft Excel spreadsheet package Microsoft Office Professional XP and program Origin 6.1. Differences between values were considered statistically significant: P < 0.05; 0.01 and 0.001.
3. Results and discussion
The immunity of animals depends on the reactivity of the organism and its main cells - T- and B-lymphocytes. T-lymphocytes and their regulatory populations are important in the formation of a specific immune response, they provide immunobiological homeostasis and resistance to anti-genic determinants (Mokrozub et al., 2012). From the data given in Table 1 we can see that the use of the investigated preparations in broiler chickens had an influence on the total number of T-lymphocytes (TE-RUL) and their functional activity. In particular, the total number of TE-RUL in the blood of broiler chickens in the experimental group at 27, 34 and 41 days of age was higher (P < 0.05-0.01) than in the control group. It should be noted that these changes were more pronounced (P < 0.001) in the blood of broiler chickens of the experimental group at the end of the experiment, in particular at 41 days of age.
Table 1
The number of TE-RUL and their functional activity in the blood of broiler chickens, % (M ± m; n = 5)
Indicators
Groups
16th day
27th day
Research periods
34th day
41th day
TE-RUL, 0 3-5 6-10
M
%
C E C E C E C E C E
52.8 ± 0.86 48.8 ± 1.31* 40.2 ± 0.96 44.0 ± 1.30* 6.6 ± 0.48 6.4 ± 0.49 0.8 ± 0,28 0.8 ± 0.25 47.65 ± 1.02 51.25 ± 1.31
49.0 ± 45.2 ± 43.6 ± 48.4 ± 6.33 ± 5.6 ± 1.83 ± 0.8 ± 51.0 ± 54.8 ±
0.89 0.86* 1.16 1.25* 0.67 0.50 0.0 0.21 0.89 0.86*
49.0 ± 0.44 43.2 ± 0.73*** 43.6 ± 0.46 51.2 ± 0.54*** 6.8 ± 0.37 5.0 ± 0.54* 0.63 ± 0.24 0.6 ± 0.24 51.0 ± 0.31 56.8 ± 1.01***
47.76 ± 0.74 40.4 ± 0.92*** 45.2 ± 0.66 52.2 ± 1.39** 6.40 ± 0.67 7.25 ± 0.48 1.0 ± 0.31 0.6 ± 0.24 52.4 ± 0.74 59.6 ± 0.93***
The increase in the total number of T-lymphocytes in the blood of broiler chickens of the experimental group occurred with a simultaneous decrease (P < 0.05-0.001) inactive functional relation of the population of TE-RUL and increase (P <0.05-0.001) the number of TE-RUL with low receptor density. These data indicate that the use of the investigated preparations led to an increase in the number of TE-RUL and increase their functional activity.
It is known that the population of blood T lymphocytes consists of several subpopulations, the cells of which differ in functional state. Therefore, the use in test of "active"
rosette formation research allows to determine the subpopulation of T-cells that have high-affinity receptors for indicator cells (erythrocytes) and actively interact with them without additional sensitization. From the data in Table 2 we see that the feeding of chickens-broilers the preparation "Bio-magn" in combination with the use of disinfectant caused an increase in blood TA-RUL. At the same time, the largest number of TA-RUL was recorded in the blood of chickens of the experimental group at 16 and 27 days of age, where differences in control were significant.
Table 2
The number of TA-RUL and their functional activity in the blood of broiler chickens, % (M ± m; n = 5)
Indicators TA-RUL 0 3-5 6-10
M
%
Research periods
16th day 27th day 34th day 41th day
C 67.2 ± 0.58 66.4 ± 1.28 66.0 ± 0.31 62.0 ± 0.31
E 65.4 ± 0.45* 61.6 ± 1.32* 60.2 ± 1.01*** 60.20 ± 0.86
C 22.2 ± 0.58 24.0 ± 2.09 28.0 ± 0.31 27.2 ± 1.95
C 26.8 ± 1.71 28.0 ± 0.83 32.0 ± 0.94** 26.6 ± 1.77
C 9.0 ± 1.18 8.2 ± 1.11 5.0 ± 0.44 10.0 ± 1.89
E 6.4 ± 1.28 9.4 ± 1.50 6.6 ± 0.4 11.8 ± 1.52
C 1.6 ± 0.49 1.4 ± 0.50 1.0 ± 0.44 0.8 ± 0.37
E 1.2 ± 0,37 1.2 ± 0.37 1.2 ± 0.37 1.4 ± 0.24
C 32.8 ± 0.58 33.6 ± 1.28 34.0 ± 0.31 38.0 ± 0.31
E 34.6 ± 0.4* 38.6 ± 1.28* 39.4 ± 1.19** 39.8 ± 0.86
The increase in the number of TA-RUL in the blood of chickens of the experimental group occurred against the background of a decrease (P < 0.05-0.001) in the number of "zero", inactive T-lymphocytes, and an increase in subpopulations with low receptor density, especially at 34 days of age (Table 2).
Similar changes were fixed in the blood of poultry in the research of the number of theophylline-resistant populations of T-lymphocytes (Table 3). Thus, in all search periods, the total number of Th - RUL in the blood of broiler chickens in
the experimental group was higher (P < 0.05-0.001) than in the control. The increase in the number of theophylline-resistant T-lymphocyte populations in the blood of chickens of the experimental group occurred with an increase (P < 0.05) in the number of low-avid and medium-avid subpopulations and a decrease (P < 0.05-0.001) in the number of inactive Th-RUL. The action of the investigated preparation showed a tendency to increase the number of T-suppressors in the blood of chickens of the experimental group in all periods of the research, and especially at 41 days of age.
Table 3
The amount of Th-RUL and Ts and their functional activity in the blood of broiler chickens, % (M ± m; n = 5)
Indicators Th, RUL, 0 3-5 6-10
M
%
Ts, % IPI
Research periods
Groups 16th day 27th day 34th day 41th day
C 62.2 ± 0.37 60.4 ± 0.50 57.4 ± 1.93 57.4 ± 0.74
E 59,0 ± 0.70 56.8 ± 0.36*** 54.8 ± 0.37 54.0 ± 0.70*
C 32.8 ± 0.8 33.6 ± 0.50 35.0 ± 1.0 35.0 ± 1.04
E 35.8 ± 0.86* 34.4 ± 0.4 39,0 ± 1.58 35.8 ± 0.86
C 4.8 ± 0.66 5.8 ± 0.37 5.4 ± 0.50 6.4 ± 0.41
E 5.2 ± 0.9 6.6 ± 0.6 5.2 ± 1.01 8.0 ± 0.31*
C 1.0 ± 0.25 0.24 ± 0.2 0.4 ± 0.24 1.2 ± 0.49
E 0 2.2 ± 0.37** 1.0 ± 0.37 1.8 ± 0.37
C 37.8 ± 0.37 39.6 ± 0.50 40.6 ± 0.60 42.6 ± 0.74
E 41.0 ± 0.70** 43.2 ± 0.37*** 45.2 ± 0.6*** 45.6 ± 0.92*
C 9.8 ± 1.15 11.2 ± 0.37 10.4 ± 0.74 9.8 ± 1.01
E 10.2 ± 0.86 11.6 ± 1.07 11.6 ± 0.67 14.0 ± 1.67
C 4.13 ± 0.59 3.54 ± 0.09 4.06 ± 0.36 4.55 ± 0.52
E 4.81 ± 0.31 3.84 ± 0.37 3.95 ± 0.27 3.45 ± 0.43
Changes in the ratio of theophylline-resistant and theo-phylline-sensitive T lymphocytes in the blood of experimental chickens led to an increase in the immunoregulatory index at 16 and 27 days and a decrease at 34 and 41 days of age, but differences in control were unlikely. However, these and previous research results indicate a positive influ-
ence of the synbiotic "Biomagn" in combination with drinking the preparation "Diolide" on the number and functional state of the receptor apparatus of T-lymphocytes.
In the analysis of the data in Table 4, attention is drawn to the probably higher number of B-lymphocytes in the blood of chickens of the experimental group on the 16th and
41th day (Table 4). In all periods of research in the blood of chickens of the experimental group, the number of EAS-RUL with low and medium density of receptors was higher,
and inactive in function - less than in poultry of the control group. At the same time, probable changes in control were fixed at 16 and 34 days of age.
Table 4
The number of B-lymphocytes and their functional activity in the blood of broiler chickens, % (M ± m; n = 5)
Indicators
Research periods
Groups 16th day 27th day 34th day 41th day
C 77.4 ± 0.65 76.60 ± 0.4 75.20 ± 0.37 74.25 ± 0.2
E 75.20 ± 0.58* 74.2 ± 0.37 73.2 ± 0.73* 72.6 ± 0.50*
C 18.8 ± 0.48 19.4 ± 0.4 20.2 ± 0.37 18.40 ± 0.93
E 20.6 ± 0.68* 21.0 ± 0.40 20.6 ± 0.40 19.8 ± 0.37
C 3.60 ± 0.50 4.0 ± 0.31 4.6 ± 0.24 6.4 ± 0.40
E 4.25 ± 0.37 4.6 ± 0.24 5.6 ± 0.24* 6.8 ± 0.37
C 0 0 0.6 ± 0.37 1.0 ± 0.47
E 0 0 0 0.8 ± 0.37
C 22.6 ± 0.65 23.4 ± 0.4 24.8 ± 0.37 25.8 ± 0.2
E 24.8 ± 0.58* 25.8 ± 0.37 28.8 ± 0.73 27.4 ± 0.50*
EAC-RUL, 0
3-5 6-10
M
%
In general, the results of investigations can be stated about the regulatory role of synbiotic preparation in combination with a disinfectant in the functioning of T- and B-cell immune components in broiler chickens. The increase in the number and redistribution of avidity of the receptor apparatus of T and B lymphocytes in the blood of poultry of the experimental group is due to the activating influence of these preparations on the reception of immunocompetent cells, which reduces antigenic load on broilers during critical growth periods. An increase in the number of T-lymphocytes (total, active, theophylline-resistant) and B-lymphocytes was observed under the action of the investigated preparations. At the same time, higher functional activity of the investigated immunocompetent cells was revealed due to the strengthening of the receptor apparatus of T- and B-lymphocytes - increase in the number of cells with low and medium degree of avidity and decrease in inactive functional relation of T- and B-lymphocytes of blood. This activating influence on the specific link of Tand B-cellular immunity of broiler chickens can be explained by the complex additive action of the investigated means, and especially the antioxidant and immunomodulatory properties of the synbiotic preparation. The increase in the number of T and B lymphocytes in the blood of broiler chickens in the experimental group, we associate with the activating influence of the investigated synbiotic preparation on the central and peripheral immune systems of poultry. Thus, many authors note that probiotics not only normalize intestinal microcoenosis and help prevent gastrointestinal diseases in young animals, but also have an influence on other systems of animals (antioxidant, immune, endocrine, etc.) (Sazawal et al., 2006; Kukkonen et al., 2007; Akpolat et al., 2009). Their therapeutic and prophylactic effect is due to the high antagonistic activity of production strains of microorganisms against pathogenic and opportunistic microflora (not even sensitive to many antibiotics), the ability to activate macrophages and interferons by binding to their receptors, causing cytokine cascades. antibody production (Kogan & Kocher, 2007; Kim et al., 2017). In particular, according to Duc Le H. et al. (2004) well-proven preparations based on microorganisms of the genus Bacillus. It is known that vegetative cells and spores, including Bacillus subtilis, which is the basis of the investigated preparation "Biomagn", passing into the lower
intestine, stimulate immunocompetent cells and macrophages. As a result, as mentioned above, the activation of the central and peripheral parts of the poultry's immunity, which ultimately manifests itself in improving the body's immune response, especially during periods of reduced immune potential of poultry in critical periods of development.
At the same time, other components containing the investigated preparation "Biomagn" have a positive influence on immune function. In this context, the milk thistle (Sily-bum marianum) deserves attention, known for its powerful detoxifying, antioxidant and immunomodulatory properties. Milk thistle contains a rare biologically active substance -silymarin. The anti-inflammatory and anti-allergic action of silymarin is associated with its ability to inhibit lipoxygen-ase (inhibits the formation of leukotrienes, inhibits neutro-phil migration, inhibits the release of histamine from baso-philic granulocytes. However, the antioxidant properties of milk thistle are associated with the functional state of the receptor apparatus of immunocompetent cells, by optimizing metabolic homeostasis, which provides high functional properties of T- and B-lymphocytes. The activating influence of Magnesium ions on the indicators of natural and adaptive immunity, in particular the binding of lymphocytes to Ig M, the response of macrophages to lymphokines is also known.
Stable functioning of the immune system in the poultry is possible subject to the connection of all links of specific immune reactions and factors of natural immune reactivity (Starovoitova et al., 2012; Klaenhammer et al., 2012; Lesyk et al., 2022).
The positive influence of complex using of disinfectant on the immune function of chickens was stated. The researches have confirmed that the best quality for drinking water is the disinfection of organic acids, chlorine dioxide, which stabilize pH, destroy many microorganisms, improve digestion, stimulate feed consumption, prevent the deposition of limescale on the walls of water supply pipes (Yu et al., 2014; Sorlini et al., 2014).
Scientists believe that setting safe levels of immunotoxi-cants - disinfectants makes it possible to determine the criteria of effective concentrations for disinfection in the presence of animals and poultry and the possibility of water disinfection (Kovalenko et al., 2005). The authors have identified and
proven that when an animal's body fails to neutralize chemical toxins, the toxic antigen can cause a chronic local delayed-type hypersensitivity reaction. The constant production of lym-phokines by sensitized T-lymphocytes leads to the accumulation of a large number of macrophages, some of which form strands of epithelioid cells, and others merge with each other to form giant cells. Macrophages, on the surface of which toxic antigen is exposed, can die, becoming targets of autolo-gous T-killers (Kovalenko et al., 2005). Additional damage to epithelial or mucous tissues occurs as a result of non-specific cytotoxicity of lymphokine-activated macrophages, as well as the action of lymphotoxins. Local accumulation in the tissue of multiplying lymphocytes and fibroblasts in the tissue form chronic granuloma (Kovalenko et al., 2005).
The analysis of indicators of natural and adaptive immune protection proved the safety and harmlessness of working solutions of the preparation "Diolide".
Therefore, an effective measure to prevent the immuno-suppressive action of disinfectants is to comply with their scientifically sound standards for use in water treatment, followed by poultry watering.
Control and detection of microbial contamination of feed used for poultry feeding is an important factor in biosafety. Some species of Escherichia coli that do not cause disease in healthy poultry can cause severe complications when infected with immunosuppressive agents. Measures to improve the quality of preparations for the prevention of avian diseases play an important role in improving bacteriological safety and ensuring the immune function of poultry.
In this regard, it is necessary to carry out comprehensive disinfection, taking into account the water supply system and to introduce probiotic preparations into the feed (Tam et al., 2003; Orishchuk et al., 2014; Catlin et al., 2018).
4. Conclusions
The use of synbiotic preparation "Biomagn" in the ration of chickens in combination with the disinfectant "Diolide" caused a positive influence on the state of T- and B-cell units of specific protection of the organism. In particular, there was an increase in the number of T-lymphocytes (total, active and theophylline-resistant) and B-lymphocytes and increase their functional activity due to the redistribution of the receptor apparatus of immunocompetent cells, which contributed to the growth of the immune potential of the poultry.
Prospects for further research. Continuation of research to study the effectiveness of new probiotics in poultry breeding of aerosol using, in order to increase the natural resistance and productivity of poultry.
Conflict of interest.
The authors state that there is no conflict of interest.
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