Study of the Effect of a Complex of Preparations Containing Aloe Arborescens Mill Extract on the Quality and Quantity of Sperm

INTRODUCTION

In recent decades, there has been an increase in the interest in practical medicine in medicines obtained from raw materials of animal and plant origin – biostimulants and adaptogens [1-4]. 

Biostimulants have been used since the time of Hippocrates, but their thorough empirical verification and scientific justification of their use began only in the twentieth century. In the preparations prepared according to the method of V. P. Filatov, substances produced by living cells in the process of their vital activity in extremely unfavorable conditions have a stimulating effect on the body. As a result, substances are formed in such tissues that stimulate specific biochemical processes in the same tissues, which contribute to the preservation of the vital activity (survivability) of tissues in unfavorable conditions [5-7].

The use of strictly dosed amounts of drugs containing such substances increases the indicators of natural resistance of the body (lysozyme and bactericidal activity of blood serum, phagocytic activity of leukocytes), increases the level of morphological and biochemical parameters of blood (the number of erythrocytes, leukocytes, hematocrit, ESR, hemoglobin, total protein), improves metabolism, increases reproductive function and stimulates growth [8-11].

The liver, spleen, testes, skin, placenta, peat, estuarine mud, plantain leaves, and other components, including Aloe arborescens Milli, are used for the manufacture of such preparations.

For example, when preparing animals for the seasonal season, many specialists intensively use such drugs to stimulate the reproductive qualities of animals. This improves the quality and quantity of semen produced in males and improves the quality of eggs in females. However, there is a fairly small number of studies on exactly how a drug or a complex of drugs affects the metabolic processes in the body, and whether the means used have a hidden negative effect on the body [12, 13].  

The genus Aloe arborescens Milli is represented by perennial tropical and subtropical plants with large thick juicy leaves. Aloe arborescens Milli is a large perennial plant, reaching 10 m in height in its homeland in Africa, and leaves up to 65 cm long. The lower part of the stem turns woody and gradually gets rid of the leaves so that the rosette of leaves remains only at the top of the shoot [14, 15].

The leaves of Aloe arborescens Milli contain oxymethylanthraquinone – Aloe-emodin (about 2%) and other anthra derivatives – aloin, nataloin, hormonal-loin. In addition, anthraglycosides, dicarboxylic acids, fatty acids, aromatic acids, vitamins, enzymes, and phytoncides are isolated from the plant. The chemical composition of the leaf juice is shown in Table 1 [16].

Table 1. Chemical composition of Aloe arborescens Milli juice

The leaves of Aloe arborescens Milli are xiphoid in shape, 15-45 cm long, 2-5.5 cm wide at the base, 0.7-1.5 cm thick; concave on the upper side, convex on the lower side, glabrous, thick, fleshy, covered with a waxy coating, with a serrated edge. The leaves have a faint peculiar smell and a very bitter taste. It is allowed to lose at least 92% of weight during drying, and at least 2% of the dry residue in the juice taken from fresh leaves before canning [17, 18].

Aloe arborescens Milli is an old remedy. It was used to treat purulent wounds, ulcers in ancient Egypt and Greece more than 3000 years ago.

In the last century in Russia, Aloe arborescens Milli preparations were used only as a laxative. When applied externally, Aloe arborescens Milli preparations have a wound-healing, anti-inflammatory, and radioprotective effect [19].

For peptic ulcers of the stomach and duodenum, Aloe arborescens Milli extract is used as a means of nonspecific stimulation in the form of subcutaneous injections, a liquid extract of Aloe arborescens Milli is prescribed inside or Aloe coated tablets [20].

The purpose of the research: to study the effect of the manufactured complex preparation based on Aloe arborescens Mill extract, E-selenium, iodine, and amylodextrin on the quality of the obtained seed and testosterone production during the preparation of rams for the breeding season.

MATERIALS AND METHODS

Taking into account the research methodology of the Institute's research project number 20.80009.5107.20: "Managementul potențialului genetic ai a producțiilor animalelor de rasă reproduce ai exploit în condițiile pedoclimaterice ale Republicii Moldova", we, after consulting with specialists of Stavropol State Medical University, Rostov State Medical University,  Saratov State University of Genetics, Biotechnology, and Engineering,  Stavropol State Agrarian University and North Caucasus Federal University, conducted experiments to determine the effect of a complex of drugs containing Aloe arborescens Milli plant extract on the quality of the seed and the production of testosterone in the preparation of rams for the transition period.

2 groups were formed to conduct the experiments: experimental and control. Each group consisted of 12 male individuals (rams), aged from 18 to 36 months, healthy, with average weight indicators.

Animals of the experimental group were given 5 ml of a drug containing iodine with amylodextrin daily for 50 days with feed, 1.5 ml/ head of E-selenium, and a tissue preparation of 0.5 ml/head mixed with 1 ml of 0.5% novocaine solution were intramuscularly injected.

The tissue preparation was manufactured in the Laboratory of Biotechnologies in Reproduction and Embryo Transfer of Scientific-Practical Institute of Biotechnologies in Animal Technology and Veterinary Medicine. The leaves of the Aloe arborescens Milli plant were used for at least 2 years. The cut leaves of Aloe arborescens Milli were kept for 10-12 days in the dark at a temperature of 4-8℃, the teeth and yellowed ends were removed, cut into small pieces, and ground until a pulp formed. A threefold amount of distilled water was added and boiled for 3-2 minutes (for protein coagulation), after which it was filtered. Sodium chloride was added to the filtrate (7 g per 1 liter), boiled for 2 minutes, and filtered, the pH level was measured (batches of the drug with a pH in the range of 5.0-5.6 were taken into operation), poured into 50 ml vials and sterilized in an autoclave at 120℃ for one hour.

Animals of the control group were not prescribed or given any drugs. Before the start of the research and at the end of the experiment, semen and blood samples were taken. Animals of both groups were kept in the same conditions and on the same diet. Qualitative and quantitative analysis of semen samples was carried out using the computer program «CEROS».  The results obtained were formalized by test certificates.

RESULTS AND DISCUSSION

The statistically processed research results were summarized in Tables 1-3 below. When evaluating the results of the studies, it should be taken into account that the sexual function of rams in both groups increases as they approach the breeding season [21, 22].

Table 2 presents the results of studies of testosterone levels in the blood of rams in their preparation for a random period in a non-random season.

Table 2. Results of studies of testosterone levels in the blood of rams in their preparation for a random period in a non-random season

Having studied the results of the studies, we see that at the beginning of the experiment, the testosterone level in rams was almost the same in both groups and was in the range of 4.57 ± 0.39 ng/ml and 4.55± 0.43 ng/ml. At the end of the experiment (taking into account the fact that the sexual function of rams in both groups increases as they approach the breeding season), it was found that the amount of testosterone in the experimental and control groups increased by 39.6% and 33.81%, respectively. In the rams of the experimental group, this indicator was 7.5±0.578 ng/ml and was 0.69 ng/ml or 0.67% higher than in the animals of the control group, where the same indicator was 6.81±0.23 ng/ml.

Tables 3 and 4 present the results of studies of rams’ semen samples in their preparation for the random period in the non-random season.

Table 3. Characteristics of the volume and concentration of sperm of rams

The sexual activity and sperm production of rams in both groups increases as they approach the breeding season, therefore, when calculating the data in Table 3, this factor was necessarily taken into account for both groups of animals.

The rams of the experimental group gave from 0.9 to 1.3 ml per cage, and in the rams of the control group, the same indicator ranged from 0.7 to 0.9 ml per cage.

Analysis of the data in Table 3 allowed us to find out that the volume of ejaculate received in the experimental group increased from 0.829 ±0.086 to 1.05 ±0.122 ml, and in the control group from 0.609 ±0.067 to 0.725±0.072 ml. Consequently, in comparison with the initial volume, the volume of ejaculate received in the experimental group increased by 21% or by 0.221 ml, and in the control group by 16% or 0.116 ml, which is less by 5% compared to the experimental group. 

The concentration of spermatozoa in rams of the experimental group increased from 1.052±0.028 billion/ml to 1.58±0.126 billion/ml, and in the experimental group, this indicator increased from 1.064±0.0351 billion/ml to 1.217±0.105 billion/ml. In comparison with the initial data, it can be seen that the concentration of sperm in the ejaculate of the experimental group increased by 33.41% or 0.528 billion/ml, and the concentration of sperm in the ejaculate of the control group increased by 12.57% or 0.153 billion/ml, which is 20.84% or 0.375 billion/ml less than in the experimental group.

Table 4. Characteristics of the quality of sperm of rams