INTRODUCTION:

Despite great success in synthetic medicines, in recent years, the popularity of phytotherapy has been on the rise. Interest in natural healing substances and preparations based on them does not weaken due to the pronounced medicinal properties of medicinal plants and rapidly developing research technologies in biology, medicine, and veterinary pharmaceuticals¹.

Preparations made from medicinal plants have unique therapeutic effects, representing multicomponent complexes of biologically active substances. Due to this, they have a wider range of pharmacotherapeutic effects compared to synthetic preparations. Many plants contain chemicals that act against various pathological processes. One medicinal plant can replace several synthetic medicines and treat diseases of organs and systems, both the main and concomitant diseases. The important advantages of phytotherapy are its versatile action and polyvalence.

Unlike synthetic medicines, plant-based preparations have a mild natural (physiological) effect and a gradual but steady therapeutic effect². They have few or no contraindications. Side effects, intolerance, and drug diseases are relatively rare in plant-based preparations. Adverse reactions from plant-based preparations are 5 times less common than when using other medicines, and they have relatively low toxicity.

Due to these positive qualities, plant-based preparations are highly effective and relatively safe, as they have high biological activity. Multicomponent plant-based preparations have a more pronounced positive clinical effect and allow the user to achieve maximum therapeutic effect. Plant-based preparations are compatible with synthetic medicines and in reasonable combinations significantly increase the therapeutic effect. The advantages of plant-based preparations include their simple preparation at home, cheapness, and availability of renewable natural raw materials annually. Medicinal plants are most effective in functional disorders and mild forms of diseases when used to increase the therapeutic effect of specific therapy and during supportive treatment³.

Modern plant-based preparations meet the standards of quality, effectiveness, and safety. They are absorbed as fully as possible without loading the body with ballast or toxic compounds or causing allergic reactions. They are usually well tolerated in humans and animals and have minimal side effects and contraindications. Modern technologies using highly purified plant-based preparations make it possible to produce preparations containing a strictly determined dose of the active substance, which facilitates the calculation of the physiological and therapeutic dose for the individual consumer.

Medicinal plant preparations succeed in treating many cardiovascular, oncological, infectious, respiratory, digestive, etc. diseases. Unfortunately, they are still not widely used in veterinary medicine^4,5. Plant-based preparations are effective in ophthalmic pathology. K.E. Boranbayeva proves the high therapeutic effectiveness of phyto ointment for bovine keratoconjunctivitis⁶.

Developing new medications with pronounced antimicrobial, anti-inflammatory, and immunomodulatory effects and minimal side effects is an urgent task of modern science and advanced technology. They are environmentally friendly, cost-effective, and easily accessible. The search for new plant-based preparations, a comprehensive study of their pharmacological properties, and introduction into wide veterinary practice are of great scientific and practical importance.

Respiratory diseases in young farm animals are characterized by widespread prevalence, high morbidity, mortality, and significant economic damage. The economic damage from respiratory diseases is due to death, forced slaughter, the costs of therapeutic and preventive measures, and a sharp decrease in the productivity of sick and recovering animals⁷. By prevalence, mortality, forced slaughter, and lack of weight gain, respiratory diseases prevail over others. Up to 80-100% of young animals are exposed to them, and the death of young animals in combination with forced slaughter can reach up to 55% or more, which leads to a decrease in the economic efficiency of the industry to 20-30%⁸.

Studies prove that respiratory diseases in animals occur as a result of their exposure to biogenic and abiogenic factors. Abiogenic factors include unfavorable environmental conditions and failure to comply with the keeping technology: unsatisfactory microclimate parameters, a large concentration of animals in limited production areas, failure to comply with the technology of completing specialized farms (complexes), non-compliance with preventive breaks between technological cycles, transportation, regrouping, abrupt change in feeding and keeping conditions, etc. Biogenic factors include viruses, bacteria, mycoplasmas, and fungi, the virulence of which increases with unfavorable keeping and feeding conditions and immunodeficiency⁹.

Abiogenic factors accumulate in the external environment and enter the animal body with feed, water, and exhaled air. These adverse environmental factors often affect the body in various combinations. However, the main factor in the development of pathological processes is inadequate animal feeding, which hurts the general condition and is accompanied by decreased productivity, natural resistance, and immuno-biological reactivity¹⁰.

There are many schemes and pharmacological agents to treat respiratory diseases of animals, including broad-spectrum antibiotics, groups of sulfonamide preparations, fluoroquinolones, etc. The main disadvantage of antimicrobial agents is that prolonged use of the same medications often leads to resistant strains of microorganisms. The irrational use of antibiotics contributes to immunity blocking, dysbiosis, and increased microorganism resistance. This decreases therapeutic effectiveness, complicating the treatment and increasing its cost¹¹.

For animal husbandry, it is necessary to introduce more effective and safe veterinary medicines, including for respiratory diseases in young animals.

Plant-based preparations from medicinal plants are increasingly used in medical and veterinary practice. Establishing the mechanism of action of biologically active medicinal substances is important from theoretical and practical points of view. A clear understanding of this allows using medicines more efficiently and charting a path to more advanced preparations.

Well-known methods of treatment and prevention of respiratory diseases of lambs are ineffective. This prompted us to improve existing measures and seek more effective means.

To expand the arsenal of medicines available for use in veterinary medicine, we developed new environmentally friendly plant-based preparations with immunomodulatory, therapeutic, and prophylactic activity for the respiratory diseases of lambs¹². We selected medicinal herbs with a pronounced therapeutic and preventive effect in diseases associated with immune system disorders and gastrointestinal and respiratory diseases.

The widespread use of preparations from plant raw materials is important for veterinary medicine since such preparations have a wider range of effects, fewer side effects, and lower risks of interaction with other medicines. They are much cheaper than synthetic medicines, are environmentally safe, and can successfully replace synthetic agents.

The study aimed to analyze the effect of plant-based preparations on the immunomorphological and microbiological parameters of lambs with respiratory diseases and to determine their therapeutic and preventive effectiveness.

MATERIALS AND METHODS:

To manufacture new plant-based preparations, we selected medicinal plants with antibacterial, antiviral, immunostimulating, therapeutic, and prophylactic effects in diseases associated with impaired immune system function and respiratory diseases: St. John's wort leaves (herbae Hypericum), sage flowers and leaves (fol. Salvia), scabwort root (rad. Inula), and marshmallow roots (rad. Althaeae) (Figures 1-4).

Figure 1. St. John's wort leaves (herbae Hypericum)

The leaves are used for medicinal purposes. The chemical composition contains 13% astringents, 1% flavonoids (hyperoside, rutin, quercitrin, myricetin, leucoanthocyanin), saponins, essential oil (0.2-0.3%), resinous substances (17%), carotene, ascorbic acid (140 mg), vitamin PP (nicotinic acid), and phytoncides.

Figure 2. Sage leaves and flowers (fol. Salvia)

Medicinal raw materials for sage are leaves and tops of plants with flowers. Flowers and leaves contain 2.5% essential oil, linalol, acetic acid, aromatic resins, pinene, formic acid, flavonoids, astringents, and phytoncides. The plant has an antiseptic, anti-inflammatory, and astringent effect.

Figure 3. Scabwort root and rhizome (rad. Inula)

The roots and rhizomes contain inulin (up to 44%), polysaccharides, resins, traces of alkaloids, saponins, essential oil (up to 4.3%), ascorbic acid, bitter substances, flavonoids, vitamin E, and zoquercitrin and quercitrin. The plant has antiseptic, anti-inflammatory, and expectorant effects.

Figure 4. Marshmallow roots (rad. Althaeae)

Marshmallow root contains starch (up to 37%), mucous substances (up to 35%), pectin (11-16%), sugar (8%), carotene, lecithin, phytosterol, mineral salts and fatty acids (1-1.5%), amino acids (from 2 to 19.8% asparagine and 4% betaine), essential oil, ascorbic acid, carotene. It has an anti-inflammatory, expectorant, and astringent effect.

Water extracts (infusions, decoctions) and alcohol extracts were prepared from crushed medicinal plants following the requirements of the State Pharmacopoeia and according to methodological recommendations. The crushed plant raw materials were placed in a glass vessel, filled with alcohol, closed, and kept in a dark place at room temperature (15-20°C) for 7 days. Then the tincture was drained, and the medicinal raw materials were squeezed, filtered, and poured into a dark bottle. Before use, the preparations were diluted with boiled water cooled to 37°C until a 10% solution was obtained.

We studied antimicrobial activity against pathogenic microorganisms using serial dilutions in a liquid nutrient medium. Strains of gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria were used as test cultures. To control the experimental preparations, we seeded the same microorganisms on nutrient media that did not contain the tested preparations. The control and experimental cultures were kept for one day in a thermostat at 37°C, after which the results were recorded. The effectiveness of the plant-based preparations was assessed by the presence of colony growth of the original microorganisms visually and by microscopy of gram-stained smears.

We used 40kg of plant raw materials, 40 heads of white mice, 20 heads of rats, 10 heads of guinea pigs, 360 heads of lambs, and Streptococcus pyogenes, S. aureus, and E. coli strains.

The scientific and production experiments were conducted at farms in the Almaty region, Kazakhstan, which usually show unfavorable results concerning respiratory diseases of viral and bacterial etiology in young animals with an incidence rate of 80% and a mortality rate of 17-20%. Preventive effectiveness was tested on clinically healthy lambs, and therapeutic effectiveness was tested on lambs aged 10-30 days with respiratory diseases. The animals were divided into experimental (EG) and control groups (CG), selected according to the principle of analogs (age, body weight, fatness). To determine the preventive effectiveness, healthy EG lambs were given infusions and decoctions of medicinal plants in a dose of 5.0 ml/kg of live weight twice a day for 10 days 20-30 minutes before feeding. To determine the therapeutic effectiveness, sick EG lambs were given alcohol medicinal plant extracts in a dose of 2.5ml/kg of live weight 20-30 minutes before feeding twice a day for 10 days. Before use, the extracts were diluted with boiled water and cooled to 37°C until a 5% solution was obtained in a ratio of 1:10. CG lambs were treated using traditional methods adopted on farms.

During the experimental period, animal health was constantly monitored, and general clinical examination methods were used, including palpation, percussion, auscultation, and thermometry. The timing of clinical recovery and the number of fallen, slaughtered, and recovered animals were considered.

To identify the causes of high morbidity, we studied nonspecific resistance and immunological reactivity in healthy and sick lambs with acute respiratory diseases. For this purpose, two groups were formed: EG1 included clinically healthy lambs, and EG2 included lambs with pronounced signs of acute respiratory diseases.

To identify the preventive and therapeutic effectiveness of the plant-based preparations, we formed six groups: a CG and five EG.

To assess the immune status of healthy and sick lambs, the absolute number of leukocytes, the relative and absolute number of lymphocytes and their main populations (T and B lymphocytes), the functional activity of neutrophils, the content of G and M immunoglobulins (IgG and IgM), lysozyme, and bactericidal activity were determined in the blood. The total blood count was determined using a Sistemex-21 blood analyzer (Japan); the content of immunoglobulins was determined using an automated Immunlight 1000 analyzer (USA); and the phagocytic activity of neutrophils was determined using KarlZeys and Prima Star microscopes (Germany) and M-50 microscopes (Austria).

The obtained digital data were processed using the constant method of variational statistics with the calculation of arithmetic averages and their statistical errors (M±m). The reliability (P) of the compared indicators was determined according to Student's t-test. The Microsoft Exel statistical analysis package was used for calculations.

RESULTS:

Clinical trials were conducted on lambs with catarrhal bronchitis. The lambs were divided into two groups (CG and EG), 10 heads each. EG lambs were given 30-50 ml of the plant-based preparation twice a day mixed with milk using a rubber bottle. CG lambs were injected with the antibiotic cefazolin. Clinical status studies were conducted before and on the 5th, 10th, and 15th day after the use of the preparations.

Before prescribing the preparations, clinical indicators in the CG and EG significantly exceeded physiological norms (Figure 5).

Figure 5. Clinical indicators of lambs with catarrhal bronchitis within 15 days from the start of the treatment in the CG and EG

EG lambs had tachycardia, rapid breathing, and palpitations. On the 5th and 10th days, clinical indicators in the EG began to stabilize better compared to the CG. Thus, the body temperature on the 5th day in the EG was 39.5±0.6°C and in the CG 40.2±0.9°C; on the 10th day, it was 38.6±0.7 and 39.0±0.9°C, respectively. The heart rate on the 5th and 10th days was 109±6.5 and 98±5.2beats/minute in the EG and 113±5.6 and 103±4.8beats/minute in the CG. In the first 5 days of the disease, the lambs had rapid breathing; the respiratory rate exceeded the physiological norm by almost twice. On the 5th and 10th days, the number of respiratory movements in the EG was 42±3.1 and 35±2.6times/minute, and in the CG 46±3.5 and 38±3.8 times/minute.

By the 15th day, in the EG, the cough and wheezing in the lungs and trachea had completely disappeared, and the clinical indicators had recovered to physiological norms, i.e. the lambs completely recovered. In the CG, the recovery was delayed due to incomplete or prolonged recovery of clinical parameters.

We studied antimicrobial activity against pathogenic microorganisms using serial dilutions in a liquid nutrient medium (Table 1).

Our studies established more pronounced bactericidal properties of extracts in various dilutions. For St. John's wort leaves, these properties were evident in dilutions of 1:40-1:80; for marshmallow rhizomes, in dilutions of 1:80-1:160; and for sage flowers and scabwort root and rhizome, in dilutions of 1:40-1:160. Bacteriostatic properties of extracts from St. John's wort leaves were evident in dilutions of 1:80-1:160; in sage flowers and scabwort rhizomes, they were evident in dilutions of 1:80-1:320; and in marshmallow rhizomes, in dilutions of 1:160-1:320.

The results of the comparative study of the antimicrobial activity of 40 and 70% alcohol tinctures are presented in Table 2.

70% alcohol tinctures of the medicinal plants had pronounced antimicrobial activity against the bacterial strains. 40% alcohol tinctures did not show antibacterial activity or showed it only slightly. In the plants, bacteriostatic activity ranged from low (1/10) to promising (1/80).

Table 1. Antimicrobial activity of extracts from various medicinal plants

No.	Medicinal plant extracts	Pathogenic tests of causative agents
		*S. pyogenes*		*S. aureus*		*E. coli*
		B/cid properties	B/stat properties	B/cid properties	B/stat properties	B/cid properties	B/stat properties
1	Folium Salvia (Sage flowers)	1:160	1:320	1:80	1:320	1:40	1:80
2	Radix Inula (Scabwort root)	1:160	1:320	1:80	1:160	1:40	1:80
3	Herbae Hypericum (St. John's wort leaves)	1:80	1:160	1:80	1:160	1:40	1:80
4	Radix Althaeae (Marshmallow root)	1:80	1:320	1:80	1:160	1:80	1:160

Note: B/cid=bactericidal properties; B/stat=bacteriostatic properties

Table 2. Antibacterial activity of tinctures (1:5) made from various medicinal plants

No.	Plant	Concentration of alcohol, %	Minimum suppressive concentration
			*S. aureus*		*E. coli*
			control	experiment	control	experiment
1	Folium Salvia (sage flowers)	40	-//-	-//-	-//-	-//-
1	Folium Salvia (sage flowers)	70	1/10	1/40	1/10	1/20
2	Radix Inula (scabwort root)	40	-//-	-//-	-//-	-//-
2	Radix Inula (scabwort root)	70	1/10	1/80	1/10	1/40
3	Herbae Hypericum (St. John's wort leaves)	40	-//-	-//-	-//-	-//-
3	Herbae Hypericum (St. John's wort leaves)	70	1/10	1/80	1/10	1/40
4	Radix Althaeae (marshmallow root)	40	-//-	-//-	-//-	-//-
4	Radix Althaeae (marshmallow root)	70	1/10	1/40	1/10	1/20

Note: "-//-" means the growth of microorganisms (lack of activity)

The most pronounced antibacterial effect was shown by alcohol extracts from St. John's wort leaves and scabwort rhizome against gram-positive microorganisms (S. aureus) (from 1/10 to 1/80). When used against gram-negative (E. coli) microorganisms, the tinctures showed a lower effect (from 1/10 to 1/40).

A comparative study of the antimicrobial activity of the plants showed that tinctures from St. John's wort leaves and scabwort rhizome had a more pronounced bacterial effect against S. aureus (1/80) and E. coli (1/40). A high level of antibacterial activity was noted in alcoholic extracts from sage flowers and marshmallow rhizome (1/20 and 1/40, respectively). The bacteriostatic activity of 70% alcohol tinctures was due to the action of the extraction solvent.

The next stage was the study of the dynamics of immunological and morphological parameters of blood. Any pathological process is accompanied by deviations in the immunological parameters. The results of studies on the immunological and morphological parameters of healthy and sick lambs are presented in Table 3.

Based on Table 3, in sick lambs with respiratory pathology, we established suppression of the immune status, accompanied by a decrease in the absolute content of leukocytes by 14.1%, lymphocytes by 19.2%, relative and absolute content of T-lymphocytes by 47.6 and 33.3% and B-lymphocytes by 26.4 and 30.2%; inhibition of the functional activity of neutrophils in spontaneous and induced tests increased by 23.2 and 50.9% compared with clinically healthy lambs (^xP<0.05; ^xxP<0.01; ^xxxP<0.001).

The concentrations of IgG and IgM also tended to decrease in CG lambs by 30.1 and 25%. The lysozyme activity was reduced by 38.8%, and the bactericidal activity was reduced by 36.9% (^xP<0.01).

Table 3. Immunological and morphological parameters of healthy and sick lambs with respiratory diseases aged 1 and 2 months (M±m; n=10)

Indicators	Animal groups
Indicators	Healthy, n=5	Sick, n=5
Leukocytes, 10⁹/l	7.44±0.32	6.52±0.45
Lymphocytes, %	52.8±2.21	44.3±2.30
Lymphocytes, 10⁹/l	5.28±0.36^x	4.47±0.29
T-lymphocytes, %	28.2±0.54^xxx	19.1±0.42
T-lymphocytes, 10⁹/l	1.6±0.1^x	1.2±0.1
B-lymphocytes, %	8.19±0.3^xxx	6.48±0.4
B-lymphocytes, 10/l	0.56±0.02^xxx	0.43±0.03
FABN, % - spontaneous test	6.84±0.3^xx	5.55±0.2
- induced test	23.1±0.69^xx	15.3±0.62
IgG, mg/ml	10.2±0.39	7.8±0.32
IgM, mg/ml	1.0±0.08	0.8±0.05
LABS, %	2.11±0.3	1.52±0.4
BABS, %	84.3±2.7^xxx	61.6±2.3

Note: ^xP<0.05; ^xxP<0.01; ^xxxP<0.001 is reliability; FABN is the functional activity of blood neutrophils; LABS is the lysozyme activity of blood serum; BABS is the bactericidal activity of blood serum

Thus, both in healthy and, to a greater extent, sick lambs, we noted a pronounced immunodeficiency condition. It had a decisive influence on the disease occurrence, development, and severity.

To control respiratory diseases, it is important to develop highly effective and accessible immune system correction tools and methods for their use in clinical veterinary practice.

A multicomponent extract from medicinal plants in a dose of 2.5ml/kg of live weight significantly activated immunological and morphological blood parameters in lambs with acute bronchitis. The obtained results are presented in Table 4.

Table 4. The effect of the multicomponent extract on the immunological and morphological blood parameters in lambs with respiratory diseases (M±m; n=10)

Indicators	Groups
	EG			CG
	Days of the study			Days of the study
	1	7	14	1	7	14
Leukocytes, 10⁹/l	6.52±0.45^x	7.69±0.50	8.56±0.52	6.52±0.45	6.59±0.59	6.91±0.52
Lymphocytes, %	44.3±2.30	53.2±2.18	56.9±2.29	44.3±2.30	45.3±1.62	48.2±1.39
Lymphocytes, 10⁹/l	4.47±0.29	5.38±0.35	5.96±0.28	4.47±0.29	4.71±0.49	4.92±0.48
T-lymphocytes, %	19.1±0.42	26.5±0.61	29.5±0.73	19.1 0.42	19.9 0.52	21.5 0.45
T-lymphocytes, 10⁹/l	1.2±0.1	1.5±0.1	1.8±0.1^xxx	1.2±0.1	1.2±0.1	1.2±0.1^xx
B-lymphocytes, %	6.48±0.49	7.89±0.35	8.71±0.51	6.48±0.48	6.59±0.46	6.88±0.43
B-lymphocytes, 10⁹/l	0.43±0.03	0.56±0.03	0.63±0.03	0.43±0.03	0.43±0.01	0.46±0.01
FABN, % - spontaneous test	5.55±0.20	6.95±0.25	7.62±0.22	5.55±0.28	5.69±0.21	5.81±0.23
- induced test	15.3±0.62	20.1±0.54	24.2±0.66	15.3±0.62	15.8±0.52	17.8±0.46
IgG, mg/ml	7.8±0.32^x	10.5±0.29	11.8±0.31	7.8±0.32	7.9±0.31^xx	8.3±0.20
IgM, mg/ml	0.8±0.05^x	1.1±0.05	1.2±0.05^xx	0.8±0.05	0.8±0.05	0.9±0.05
LABS, %	1.52±0.4	2.21±0.4	2.39±0.4	1.52±0.4	1.62±0.4	1.83±0.4
BABS, %	61.6±2.3	79.5±2.1	86.2±2.4	61.6±2.1	62.8±2.5	67.9±2.3

The immunological and morphological parameters in EG lambs significantly exceeded the initial data. Thus, the concentration of leukocytes on the 7th and 14th days increased by 17.9 and 31.3% compared to the baseline indicator. A similar upward trend was noted for lymphocytes (20.1 and 28.4%). The largest increases were recorded for T-lymphocytes (38.7 and 54.5%) and B-lymphocytes (21.8 and 34.4%).

Higher indices were noted in the FABN. The FABN level increased by 25.2 and 37.3% in the spontaneous test and by 31.3 and 58.2% in the induced test compared with the initial data (^xP<0.05; ^xxP<0.01; ^xxxP<0.001).

The results indicate that the multicomponent plant extract activated the immunoglobulin composition. Thus, the concentration of IgG increased by 36.6 and 51.3% and the concentration of IgM by 37.5 and 50.0% compared to the initial data (^xP<0.05; ^xxP<0.01; ^xxxP<0.001).

Humoral immune protection factors were also significantly activated under the influence of the plant extract. Thus, the level of lysozyme activity increased by 45.4 and 57.2% and the level of bactericidal activity increased by 29.1 and 39.9% compared with the baseline indicator (^xP<0.05).

The immunological and morphological parameters compared with the CG also tended to increase. Thus, in the EG compared with the CG on the 7th and 14th days, the number of leukocytes increased by 16.7 and 23.9%; T-lymphocyte concentrations by 19.7 and 37.2%; B-lymphocytes by 19.7 and 26.6% (^xP<0.05; ^xxP<0.01).

Under the influence of the plant-based preparation, the FABN increased; the degree of its increase in the EG in the spontaneous test was 22.1 and 31.2%, and in the induced test 27.2 and 36.0%.

A significant increase in the EG was noted for the quantitative content of immunoglobulins. Thus, the increase in IgG concentration compared with the CG averaged 37.6%. IgM increased on average by 35.4% (^xP<0.05; ^xxP<0.01; ^xxxP<0.001).

The humoral indicators of nonspecific resistance in the EG significantly exceeded those of the CG. Thus, the degree of increase in the level of lysozyme activity in the EG was up to 2.21±0.4 and 2.39±0.42.39±0.4%, while in the CG, the indicators were 1.62±0.4 and 1.83±0.41.83±0.4% (^xP<0.05; ^xxP<0.01; ^xxxP<0.001).

Having observed the beneficial effects of the plant-based preparation from the mixture of medicinal plants on clinical, microbiological, immunological, and morphological blood parameters, we set the task to study their therapeutic and preventive effectiveness in production.

The data indicate that the preventive effectiveness of the alcohol-based extracts significantly exceeded that of the aqueous extracts. The preventive effectiveness of the plant extracts individually amounted to 93.3%. The greatest effectiveness was obtained from the complex plant-based preparation with the survivability of lambs and the effectiveness reaching 100%.

During the experiment, 6 (5%) out of 120 healthy lambs from EG1-4 fell ill with respiratory diseases; four sick lambs (3.33%) died, while 96.7% survived. Absolute preventive effectiveness was obtained from the complex preparation with 100% effectiveness and survivability. Of the 30 CG heads, the number of cases amounted to 7 (23.3%), of which 5 (16.67%) died despite the medical measures. In the CG, the survivability was 83.3%, and the preventive effectiveness was only 76.7%. All the medicinal forms of plant-based preparations provided milder forms of the disease in the EG.

Our results indicate a higher therapeutic effectiveness of medicinal plant extracts used individually and as a complex preparation (Table 5).

The therapeutic effectiveness of St. John's wort leaf extract (EG1) was 93.3%, the number of deaths was 6.67%, relapses were registered in 1 lamb (3.33%), the average daily live weight gain was 209.0±14.5 g, and the absolute live weight gain for 3 months was 19.04±0.79 kg. These indicators were higher than in the CG by 21 g and 1.93 kg. Similar results were obtained using scabwort rhizome extract (EG2), where the therapeutic effectiveness also amounted to 93.3%.

Absolute 100% therapeutic effectiveness was observed with the complex preparation (EG3). Here, relapses and deaths were not recorded. The preparation reduced the duration of the disease by an average of 3-5 days compared to the CG. The average daily live weight gain was greater by 15.2% and the absolute live weight gain for 3 months by 15.9% than in the CG (^xP<0.05).

DISCUSSION:

The pronounced bacteriocidal and bacteriostatic effects of the tested plant extracts are due to the various components in their chemical composition, in particular, essential oils, glycosides, alkaloids, tannins, and ascorbic acid. 70% alcohol tinctures of the medicinal plants had pronounced antimicrobial activity against the bacterial strains. 40% alcohol tinctures did not show antibacterial activity or showed it only slightly. The most pronounced antibacterial effect was shown by alcohol extracts from St. John's wort leaves and scabwort rhizome against gram-positive microorganisms (S. aureus) (from 1/10 to 1/80). When used against gram-negative (E. coli) microorganisms, the tinctures showed a lower effect (from 1/10 to 1/40). Our results are consistent with other research.

Any pathological process is accompanied by deviations in the immunological and morphological parameters. Among the methods to objectively assess the health and the course of the pathological process, a special place is occupied by blood testing. Blood performs an important function providing optimal conditions for the normal functioning of organs and systems. In various conditions, immunological and morphological indicators can decrease, and immunodeficiency can develop, leading to an imbalance in the immune system.

In sick lambs with respiratory pathology, we observed suppression of the immune status, accompanied by a decrease in the absolute content of leukocytes and lymphocytes and inhibition of the functional activity of blood neutrophils, IgG and IgM, lysozyme, and bactericidal activity in the EG and CG, which indicates immunodeficiency.

Developing highly effective and affordable medicinal products from medicinal plants for veterinary practice is important to improve measures against respiratory diseases.

We showed that the immunological and morphological parameters in the EG significantly exceeded those of the CG. Under the influence of the extracts, morphological parameters of blood (leukocytes, lymphocytes), humoral links of immune protection (lysozyme and bactericidal activity, IgG and IgM), and cellular links of nonspecific resistance (T-lymphocytes, B-lymphocytes, FABN) were significantly activated.

An increase in the quantitative content of immunological and morphological indicators also characterizes the activation of the immunological reactivity, since they provide a better supply of oxygen to tissues, increase the intensity of metabolic reactions, and contribute to a faster recovery. Quantitative and qualitative changes in the immunological and morphological parameters of blood are also reported by many researchers¹³.

The immunoreactive state of the body is assessed based on indicators of humoral and cellular immunity. Immunoglobulins are components of humoral immunity. The quantitative composition of immunoglobulins allows us to assess the relationship between the pathological process and the state of resistance of the body. Our data on serum immunoglobulins when using various preparations are consistent with other scientific works.

An increase in the number of immunoglobulins indicates the immune correcting effect of the plant-based preparation, which has a positive effect on the course of the disease.

The state of the immune system is the main indicator of the impact of negative environmental factors and serves as a criterion for the risk of developing diseases. The modern detection of immunodeficiency and its correction to the physiological norm is an important link to increased nonspecific resistance. One of the main criteria for evaluating cellular factors of body protection is the phagocytic activity of blood cells since high resistance to adverse factors is largely provided by the phagocytic activity of neutrophils, which is the dominant link in the system of natural resistance.

To assess the physiological state, the number of leukocytes is of great interest. The level of white blood cells characterizes the activity of protective reactions associated with an inflammatory process aimed at compensating for the deficiency of cellular and humoral protective factors.

A low level of humoral and cellular mechanisms of immune protection serves as a predisposing criterion for respiratory diseases. Using plant-based preparations, the activity of phagocytic blood cells increases, and they play a decisive role in the system of natural resistance and stabilization of immunity.

Our data indicate that the medicinal plant preparations have a pronounced therapeutic and prophylactic effect. They also have an immune corrective effect on the humoral and cellular mechanisms of natural resistance, providing the conditions for rapid recovery. Our research is consistent with those of Sh.B. Turzhigitova¹⁴, who conducted scientific experiments on calves using plant-based preparations.

Our results confirm the higher preventive and therapeutic effectiveness of medicinal plant extracts used individually and as a complex preparation. The preventive effectiveness of medicinal plant extracts used individually amounted to 93.3%. The greatest effectiveness was observed with the complex preparation, where the survivability and effectiveness equaled 100%.

The absolute therapeutic effectiveness was obtained when using the complex preparation; where no deaths or relapses were recorded. The data showed noticeable increases in the daily average and absolute live weight gain. The preparation reduced the duration of the disease by an average of 3-5 days compared to the CG. The average daily live weight gain was 15.2% higher, and the absolute live weight gain for 3 months was 15.9% higher than in the CG.

CONCLUSIONS:

Our data indicate that the lambs had immunodeficiency accompanied by the immune system imbalance. Positive results were obtained when using plant-based preparations for the correction of the immunological parameters, confirmed by a significant increase in the level of immunological and morphological parameters.

The preparation from medicinal plant raw materials significantly reduced the risk of morbidity in lambs with respiratory pathology, with 100% survivability and a higher live weight gain of animals compared with the CG. The results give grounds to recommend new dosage forms of medicinal plants as agents that increase the effectiveness of traditional therapeutic and preventive measures for respiratory diseases in lambs.

We recommend using the complex plant-based preparation as an effective preventive and therapeutic agent. The preventive effectiveness of the complex preparation was 96.7% against 80% in the CG. The therapeutic effectiveness of aqueous extractions was 96.7%; in alcohol extractions, it equaled 100%.

The average daily and absolute live weight gain in the EG was on average 15-16% higher than in the CG. The complex preparation helped to increase the immune status, ensuring high preventive and therapeutic effectiveness and survivability in lambs with massive respiratory diseases.

One limitation of our study is the small sample size, which reduces the generalizability of the results. The study was conducted in Kazakhstan, and the findings may not apply to other regions with different environmental conditions and livestock management practices.

Future research should evaluate the effectiveness of these treatments against a wider range of pathogens and work towards standardizing the preparation and dosage of plant-based reparations for consistent results.

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Received on 18.07.2024 Revised on 16.08.2024

Accepted on 20.09.2024 Published on 20.01.2025

Available online from January 27, 2025

Research J. Pharmacy and Technology. 2025;18(1):295-304.

DOI: 10.52711/0974-360X.2025.00046

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Indicators	Animal groups
Indicators	EG1 (St. John's wort extract)	EG2 (scabwort extract)	EG3 (complex plant-based preparation)	CG
Number of animals, heads	30	30	30	30
Recovered animals, heads	28	28	30	24
Deaths, heads	2	2	-	6
in %	6.67	6.67	-	20
Duration of the disease, days	6-8	6-8	6-7	9-12
Therapeutic effectiveness, %	93.3	93.7	100	80
Relapses, heads	1	1	-	5
in %	3.33	3.33	-	16.7
Average daily live weight gain, g	209.0±15.1	209.5±13.8	215.5±16.3	187.0±15.6
Absolute live weight gain for 3 months, kg	19.23±0.79	19.27±0.91	19.92±0.94	17.19±0.79