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Evaluation of the antibacterial effect of Artemisia lerchiana compared with various medicines

Avaliação do efeito antibacteriano da Artemisia lerchiana comparado a outros medicamentos

Abstract

Many antimicrobial substances, mainly of chemical origin, are not effective and reliable. Many of them tend to accumulate in organs and tissues and have allergic and toxicological effects and side effects. Therefore, the purpose of our research was to conduct a comparative analysis of the antibacterial effect of Artemisia lerchiana against other herbal preparations. A. lerchiana was used in the form of an infusion and essential oil extract to fulfill the set goal and objectives. For comparative analysis, tinctures of A. absinthium, Calendula officinalis L., and Chamomilla recutita L., as well as preparations based on Abies Sibirica, Eucalyptus, Limon, Bergamottae, Melaleuca alternifolia, and A. absinthium essential oils were used. As a test standard for comparing antibacterial properties, we used the Septarius chemical preparation manufactured at a biofactory. The experiment was carried out in vitro using test microbes of gram-positive Staphylococcus aureus and Streptococcus and gram-negative Escherichia coli. The results of the study showed that A. lerchiana had a high antibacterial effect against gram-positive test microbes (Staphylococcus, Streptococcus). The most effective form was A. lerchiana in the form of essential oil, which had an antibacterial effect even in low concentrations. A comparative assessment of A. lerchiana with the Septarius test antiseptic showed that A. lerchiana had a similar effect in general. In the form of essential oil, it even surpassed the effect of Septarius. It was found that A. lerchiana surpassed medicinal plants used in the experiment in terms of antibacterial effect. The obtained results of the study will be useful for scientists researching various medicinal products of plant origin and for veterinary specialists looking for therapeutic and preventive measures.

Keywords:
Artemisia lerchiana; microorganisms; antimicrobial effect; essential oil; infusion

Resumo

Muitas substâncias antimicrobianas, principalmente de origem química, não são eficazes e confiáveis. Muitos deles tendem a se acumular em órgãos e tecidos e apresentam efeitos alérgicos e toxicológicos e efeitos colaterais. Portanto, o objetivo desta pesquisa foi realizar uma análise comparativa do efeito antibacteriano da Artemisia lerchiana contra outras preparações fitoterápicas. A. lerchiana foi utilizada na forma de infusão e extrato de óleo essencial para cumprir a meta e os objetivos traçados. Para análise comparativa, foram utilizadas tinturas de A. absinthium, Calendula officinalis L. e Chamomilla recutita L., bem como preparações à base de óleos essenciais de Abies Sibirica, Eucalyptus, Limon, Bergamottae, Melaleuca alternifolia, e A. absinthium. Como padrão de teste para comparação de propriedades antibacterianas, utilizamos a preparação química Septarius fabricada em uma Biofábrica. O experimento foi realizado in vitro utilizando micróbios de teste de Staphylococcus aureus e Streptococcus gram-positivos e Escherichia coli gram-negativa. Os resultados do estudo mostraram que A. lerchiana teve um alto efeito antibacteriano contra micróbios de teste gram-positivos (Staphylococcus, Streptococcus). A forma mais eficaz foi a A. lerchiana na forma de óleo essencial, que teve efeito antibacteriano mesmo em baixas concentrações. Uma avaliação comparativa de A. lerchiana com o antisséptico Septarius test mostrou que A. lerchiana teve um efeito semelhante, em geral. Na forma de óleo essencial, superou até o efeito do Septário. Verificou-se que A. lerchiana superou as plantas medicinais utilizadas no experimento em relação ao efeito antibacteriano. Os resultados obtidos do estudo serão úteis para cientistas que pesquisam diversos medicamentos de origem vegetal e para especialistas veterinários que buscam medidas terapêuticas e preventivas.

Palavras-chave:
Artemisia lerchiana; microrganismos; efeito antimicrobiano; óleo essencial; infusão

1. Introduction

Many antimicrobial substances, mainly of chemical origin, are not effective and reliable. Some are used as disinfectants or antiseptics for the skin and others for the prevention of infectious diseases in animals. In addition, many of them tend to accumulate in organs and tissues and have allergic and toxicological effects and side effects (Turzhigitova et al., 2022TURZHIGITOVA, S., ZAMANBEKOV, N., KORABAYEV, Y., KAZIYEV, Z. and TUGANBAY, A., 2022. Technology for obtaining dosage forms (tinctures, extracts) from local plant raw materials and studying their toxicity. Research Journal of Pharmacy and Technology, vol. 15, no. 8, pp. 3540-3548. http://dx.doi.org/10.52711/0974-360X.2022.00594.
http://dx.doi.org/10.52711/0974-360X.202...
).

In recent years, scientific studies have confirmed the growth of microorganism strains resistant to all classes of chemical compounds (Temreshev et al., 2022TEMRESHEV, I., TURSYNKULOV, A., DUTBAYEV, Y., MAKEZHANOV, A. and SULEIMANOVA, G., 2022. Evaluation of the efficacy of Entolek K Planteco® biopesticide based on akanthomyces lecanii fungus against pest orthopterans in soybean agroecosystems in Southeast Kazakhstan. Online Journal of Biological Sciences, vol. 22, no. 4, pp. 502-511. http://dx.doi.org/10.3844/ojbsci.2022.502.511.
http://dx.doi.org/10.3844/ojbsci.2022.50...
). The absence of a unified scientific concept of microorganism control based on the laws of biology, physics, and chemistry does not make it possible to achieve any significant success in the sanitary and epidemiological protection of the population and animals (Bakhir et al., 2003BAKHIR, V.M., VTORENKO, V.I., LEONOV, B.I., PANICHEVA, S.A., PRILUTSKII, V.I. and SHOMOVSKAYA, N.Y., 2003. Efficacy and safety of chemicals for disinfection, preventive treatment and sterilization. Dezinfektsionnoe Delo, no. 1, pp. 57-62.; Fraise et al., 2013FRAISE, A.P., MAILLARD, J.-Y. and SATTAR, S., 2013. Russel, Hugo & Ayliffe’s principles and practice of disinfection, presservation and sterilization. 5th ed. Chichester: Blackwell Publishing Ltd. http://dx.doi.org/10.1002/9781118425831.
http://dx.doi.org/10.1002/9781118425831...
; Nosik et al., 2004NOSIK, N.N., NOSIK, D.N., DERYABIN, P.G. and IVANOVA, E.B., 2004. Modern approaches to the study and evaluation of the virucidal activity of disinfectants. Dezinfektsionnoe Delo, no. 1, pp. 54-57.). The allergic tendencies arising in the animal's body to existing antimicrobial substances and the formation of resistance of microorganisms can be called one of the most important problems.

To date, the search for new highly effective herbal preparations without side effects is becoming relevant in solving the main issues of veterinary medicine (Boranbayeva et al., 2023BORANBAYEVA, K.E., ZAMANBEKOV, N.A., SATTAROVA, R.S., SPIRIDONOV, G.N., ZHYLGELDIYEVA, A.A. and TURZHIGITOVA, S.B., 2023. Comparative pharmacotherapeutic effectiveness of therapeutic ointments in infectious keratoconjunctivitis in cattle. Research Journal of Pharmacy and Technology, vol. 16, no. 1, pp. 46-54. http://dx.doi.org/10.52711/0974-360X.2023.00009.
http://dx.doi.org/10.52711/0974-360X.202...
; Gayirbegov and Engurazov, 2023GAYIRBEGOV, D. and ENGURAZOV, G., 2023. Effect of the cresacin feed additive on the digestibility of nutrients, productivity and egg quality in egg laying quails. American Journal of Animal and Veterinary Sciences, vol. 18, no. 1, pp. 52-57. http://dx.doi.org/10.3844/ajavsp.2023.52.57.
http://dx.doi.org/10.3844/ajavsp.2023.52...
; Korotkiy et al., 2023KOROTKIY, V.P., ZAITSEV, V.V., BOGOLYUBOVA, N.V., ZAITSEVA, L.M. and RYZHOV, V.A., 2023. The effect of a pine tree energy supplement on methane release by lactating cows. Research Journal of Pharmacy and Technology, vol. 16, no. 4, pp. 1627-1632. http://dx.doi.org/10.52711/0974-360X.2023.00266.
http://dx.doi.org/10.52711/0974-360X.202...
; Smirnova et al., 2023SMIRNOVA, I., SADANOV, A., BAIMAKHANOVA, G., FAIZULINA, E. and TATARKINA, L., 2023. Using salt-tolerant rhizobia to improve the soybean (Glycine max) Resilience to salinity. SABRAO Journal of Breeding and Genetics, vol. 55, no. 3, pp. 810-824. http://dx.doi.org/10.54910/sabrao2023.55.3.17.
http://dx.doi.org/10.54910/sabrao2023.55...
). To do this, substances with an antimicrobial effect must meet several following requirements:

  • they must have a wide spectrum of antimicrobial action, that is, effectively destroy bacteria, mycobacteria, viruses, fungi, and spores, regardless of the duration and frequency of use, ensuring that these preparations have properties that prevent the development of resistance in microorganisms;

  • the preparations should not harm humans and animals, both during their preparation and application and after the end of their purposeful use, that is, during the period of degenerative and destructive changes caused by environmental factors or biodegradation processes in the human body. In other words, antimicrobial substances and products of their natural or artificial degradation should not contain xenobiotic substances;

  • the antimicrobial substance should have a washing capacity with the least damaging and corrosive activity concerning various materials and be as easy to use and relatively cheap (Gunar et al., 2022GUNAR, O.V., DORENSKAYA, A.V., BULGAKOVA, G.M. and SAKHNO, N.G., 2022. Bactericidal and fungicidal properties of some antiseptic drugs and disinfectants. Pharmaceutical Chemistry Journal, vol. 56, no. 6, pp. 866-871. http://dx.doi.org/10.1007/s11094-022-02720-0.
    http://dx.doi.org/10.1007/s11094-022-027...
    ; Lee et al., 2023LEE, G.-H., PARK, S.-H., SONG, B.-M., KIM, D.M., HAN, H.J., PARK, J.Y., JO, Y.W., HWANG, M.Y., SIM, K.T., KANG, S.-M. and TARK, D., 2023. Comparative efficacy evaluation of disinfectants against severe acute respiratory syndrome coronavirus-2. The Journal of Hospital Infection, vol. 131, pp. 12-22. http://dx.doi.org/10.1016/j.jhin.2022.09.011. PMid:36183929.
    http://dx.doi.org/10.1016/j.jhin.2022.09...
    ).

Given the large number of commercially available herbal and natural medicines, selecting the most effective ones becomes difficult. The medicinal plant Artemisia lerchiana is of the greatest interest. In recent years, studies of antibacterial, viral, anti-inflammatory, nematocidal, and fungicidal properties of essential oils and extracts of medicinal plants have been of undoubted interest (Ćavar et al., 2012ĆAVAR, S., MAKSIMOVIĆ, M., VIDIC, D. and PARIĆ, A., 2012. Chemical composition and antioxidant and antimicrobial activity of essential oil of Artemisia annua L. from Bosnia. Industrial Crops and Products, vol. 37, no. 1, pp. 479-485. http://dx.doi.org/10.1016/j.indcrop.2011.07.024.
http://dx.doi.org/10.1016/j.indcrop.2011...
; Mamatova, 2018MAMATOVA, A.S., 2018. Pharmacognostic and pharmacotechnological treatment of Artemisia gmelinii and creation of phytosubstances based on it. Almaty: Kazakh National Medical University named after S.D. Asfendiyarov. Dissertation for the degree of Doctor of Philosophy.; Nibret and Wink, 2010NIBRET, E. and WINK, M., 2010. Volatile components of four Ethiopian Artemisia species extracts and their in vitro antitrypanosomal and cytotoxic activities. Phytomedicine, vol. 17, no. 5, pp. 369-374. http://dx.doi.org/10.1016/j.phymed.2009.07.016. PMid:19683909.
http://dx.doi.org/10.1016/j.phymed.2009....
; Verma et al., 2011VERMA, R.K., CHAUHAN, A., VERMA, R.S. and GUPTA, A.K., 2011. Influence of planting date on growth, artemisinin yield, seed and oil yield of Artemisia annua L. Under temperate climatic conditions. Industrial Crops and Products, vol. 34, no. 1, pp. 860-864. http://dx.doi.org/10.1016/j.indcrop.2011.02.004.
http://dx.doi.org/10.1016/j.indcrop.2011...
). Among the class of metabolites of the Artemisia family, terpenoids are the most studied substances. A promising field in the treatment of inflammatory diseases is the local use of plant essential oils.

Recently, medicinal plants have been of the greatest interest, as they can be not only a basis for the development of agents with antimicrobial effects but also a source of compounds with the necessary transforming activity concerning the resistance of pathogens (Badea and Delian, 2014BADEA, M.L. and DELIAN, E., 2014. In vitro antifungal activity of the essential oils from Artemisia spp. L. on Sclerotinia sclerotiorum. Romanian Biotechnological Letters, vol. 19, no. 3, pp. 9345-9352.; Ghendov et al., 2018GHENDOV, V., IZVERSCAIA, T., CIOCARLAN, N. and SIMONNET, X., 2018. Medicinal plant resources of southern arid steppes in Republic of Moldova. In: A.A. CHIBILEV, ed. Stepi severnoi Evrazii. Orenburg: Institute of Steppe of the Ural Branch of the Russian Academy of Sciences, pp. 277-280.; Lenardis et al., 2011LENARDIS, A.E., MORVILLO, C.M., GIL, A. and DE LA FUENTE, E.B., 2011. Arthropod communities related to different mixtures of oil (Glycine max L.Merr.) and essential oil (Artemisia annua L.) crops. Industrial Crops and Products, vol. 34, no. 2, pp. 1340-1347. http://dx.doi.org/10.1016/j.indcrop.2010.12.001.
http://dx.doi.org/10.1016/j.indcrop.2010...
). Recent studies have shown that the formation of biofilm by microorganisms for resistance to antibiotics, phagocytosis, and other components of the body's defense system has led to the discovery and isolation of many compounds, such as monoterpenoids, sesquiterpenoids, flavonoids, and coumarins, as well as aliphatic and lipid compounds (Bajpai et al., 2014BAJPAI, V.K., SHARMA, A. and BAEK, K.H., 2014. Antibacterial mode of action of the essential oil obtained from Chamaecyparis obtusa sawdust on the membrane integrity of selected foodborne pathogens. Food Technology and Biotechnology, vol. 52, no. 1, pp. 109-118.; Naili et al., 2010NAILI, B.M., ALGHAZEER, R.O., SALEH, N.A. and AL-NAJJAR, A.Y., 2010. Evaluation of antibacterial and antioxidant activities of Artemisia campestris (Asteraceae) and Ziziphus lotus (Rhamnacea). Arabian Journal of Chemistry, vol. 3, no. 2, pp. 79-84. http://dx.doi.org/10.1016/j.arabjc.2010.02.002.
http://dx.doi.org/10.1016/j.arabjc.2010....
; Utkina et al., 2012UTKINA, T.M., POTEKHINA, L.P., VALYSHEVA, I.V. and KARTASHOVA, O.L., 2012 [viewed 11 October 2023]. The effect of essential oils on the growth and persistence of staphylococci. Sovremennye Problemy Nauki i Obrazovaniya [online], no. 6, online. Available from: https://science-education.ru/ru/article/view?id=7360
https://science-education.ru/ru/article/...
).

A priority field is the study of the basic properties of essential oils and extracts of Artemisia growing in the Republic of Kazakhstan and the development of new medicines based on them. Kazakhstan has unique reserves of medicinal plant raw materials. 81 Artemisia species grow in Kazakhstan, and only a small part of them have been studied: A. armeniaca Lam., A. atomentella, A. annua, A. pontica, A. tournefortiana, A. laciniata, A. semiarida, A. albida, A. marschalliana), etc. (Mamatova, 2018MAMATOVA, A.S., 2018. Pharmacognostic and pharmacotechnological treatment of Artemisia gmelinii and creation of phytosubstances based on it. Almaty: Kazakh National Medical University named after S.D. Asfendiyarov. Dissertation for the degree of Doctor of Philosophy.).

Analyzing these data, we can state the prospect of studying Artemisia essential oils as active and auxiliary substances in the creation of new medicines for the treatment of infectious and inflammatory diseases associated with stable microflora. The antimicrobial effect of various Artemisia types also depends on the place of growth and extraction technology. Among the medicinal plants growing on the territory of Kazakhstan, only some Artemisia species have been studied. The Artemisia species are considered endemic species growing in the West Kazakhstan region. The determination of the antimicrobial action of biologically active substances contained in A. lerchiana emphasizes the importance of the study.

Therefore, the purpose of our research work was to conduct a comparative analysis of the antibacterial effect of A. lerchiana against other herbal preparations.

The objectives of the research included studying the antibacterial effect of A. lerchiana against gram-positive and gram-negative test microbes, as well as conducting a comparative assessment with other medicinal plants with antibacterial properties. The evaluation of the antibacterial effect was carried out in comparison with the Septarius chemical antiseptic.

2. Materials and Methods

2.1. Place and conditions of the study

To reach the set goals and objectives, we carried out experiments in the research laboratory of microbiology of the Zhangir Khan West Kazakhstan Agrarian and Technical University in equal conditions under a certain microclimate (t: 200С, relative humidity: 55%, air velocity: 0.2 m/s).

2.2. Comparative analysis

The test preparation A. lerchiana was used in the form of an infusion and essential oil extract, which had been made by generally accepted methods of pharmacology. For comparative analysis, we used the infusions of A. absinthium, Calendula officinalis L., and Chamomilla recutita L., as well as preparations based on essential oils of Abies Sibirica, Eucalyptus, Limon, Bergamottae, Melaleuca alternifolia, and A. absinthium.

As a test standard for comparing antibacterial properties, we used the Septarius chemical preparation manufactured at a biofactory.

The experiment was carried out in vitro using test microbes of gram-positive Staphylococcus aureus and Streptococcus and gram-negative Escherichia coli.

2.3. Study of antibacterial properties

To study the antibacterial properties of infusions made of various plants, we used the antibacterial effect determination method for the preparation against test microbes.

To do this, we used meat-peptone broth (MPB) in test tubes (5 ml), to which 0.3 ml of the test microbe was added (using doses of 500 million, 1 billion, and 2 billion colony-forming units (CFU)) and the test preparations in doses of 0.5, 1, and 2 ml. Subsequently, the test tubes were placed in a thermostat for 24 hours. The results of the experiments were evaluated by the growth of microbes according to the turbidity standard with calibration in percentages and cross-calibration (Table 1). The control evaluation was carried out by counting microbes under a microscope, where colonies of microorganisms were counted in four fields per 1 cm2. The calculation was carried out as a percentage relative to the turbidity standard of the test microbe.

Table 1
Test microbe growth evaluation scale.

In the control group, 1 ml of distilled water was used instead of the test preparation.

To study the antibacterial properties of essential oils of various plants, we used the disk diffuse method in the MPB. For this purpose, the MPB was used in Petri dishes, where one test microbe (S. aureus, Streptococcus, E. coli) was added. Subsequently, disks soaked in essential oils of various test plants or a Septarius solution were placed on the MPB.

After that, the Petri dishes were placed in a thermostat for 24 hours.

The results of the experiments were evaluated by determining the lysis zone in mm.

2.4. Stages of the study

The experiment was conducted in two stages. At the first stage, the antibacterial effect of A. lerchiana in various dilutions was determined (100%, 50%, 25%, 12.5%, 6.25%). In the second stage, the antibacterial effect of various plant essential oils was determined.

A comparative analysis was carried out concerning the antibacterial effect of preparations from various plants and Septarius. For reliability, all experiments were carried out in a fivefold repetition.

3. Results

Studies of the antibacterial efficacy of A. lerchiana against Staphylococcus (Table 2) showed its high effect at a 500 million CFU dose of microbes, regardless of the volume of the preparation. At a 1 billion CFU dose of microbes, the effect of the preparations remained at high volumes (1 and 2 ml), whereas in 0.5 ml of the preparation, there was a noticeable decrease in effect by 1 point (25%).

Table 2
Antibacterial effect of medicinal plant infusion against S. aureus.

With a billion CFU dose of S. aureus microbes, the effect of the test preparation is defined as 50% at 1 and 2 ml and only 25% at 0.5 ml.

Studies of the antibacterial effect of other plant infusions have shown 50% effectiveness at 500 million and 1 billion CFU, while at 2 billion CFU no antibacterial effect was observed. The worst indicators of antibacterial effect were observed in the calendula infusion.

Studies of the antibacterial effect of Septarius showed an identical result with the preparation of A. lerchiana.

Studies of the antibacterial effect of A. lerchiana against Streptococcus (Table 3) showed a high effect at 500 million and 1 billion CFU doses of microbes in 1 and 2 ml.

Table 3
Antibacterial effect of medicinal plant infusion against Streptococcus.

A decrease in the effect of A. lerchiana (50%) was observed at a 2 billion CFU dose of microbes in 1 and 2 ml. In a volume of 0.5 ml, the preparation had almost no effect at a 2 billion CFU dose of microbes.

The antibacterial effect of A. lerchiana was higher than that of Septarius at a 2 billion CFU dose of microbes.

We noted the low antibacterial effect of A. absinthum and C. recutita L. infusion at 500 million and 1 billion CFU doses of microbes. The antibacterial effect was not observed in the tested preparations at a 2 billion CFU dose of microbes.

The infusion of C. Officinalis L. proved ineffective against microbes at doses of 1 billion and 2 billion CFU and showed a 50% effect at a 500 million CFU dose of microbes.

Studies of the antibacterial effect of A. lerchiana against E. coli (Table 4) showed a high effect at a 500 million CFU dose of microbes in 1 and 2 ml, whereas a 50% effect was observed in 0.5 ml and at a 1 billion CFU dose of microbes. In 1 and 2 ml, the antibacterial effect of A. lerchiana was not observed against E. coli microbes at a dose of 2 billion CFU.

Table 4
Antibacterial effect of medicinal plant infusion against E. coli.

Septarius showed a high antibacterial effect at a dose of microbes of 500 million and 1 billion CFU in 1 and 2 ml and a 50% effect in other studies.

The infusion of C. recutita L. showed a 50% antibacterial effect at a 500 million CFU dose of microbes while in other cases, this preparation had no effect.

Besides, infusions of A. Sibirica and C. Officinalis L. also showed no effect against Escherichia coli.

In all experiments in the control group, an intensive growth of microbes was observed, which proves the high activity of test microbes.

Thus, the results of experiments show the effectiveness of A. lerchiana infusion against S. aureus, Streptococcus, and E. coli at a 500 million CFU dose of microbes. A comparison of this preparation with Septarius, as an antiseptic standard, shows the identity of the result concerning S. aureus and Streptococcus. A. lerchiana was less effective against E. coli compared to Septarius.

Comparative analysis of various herbal infusions with A. lerchiana infusion shows a high antibacterial effect of the test preparation against both gram-positive (S. aureus, Streptococcus) and gram-negative (E. coli) test microbes. The infusion of C. Officinalis L. showed a very low antibacterial effect, while the infusions of A. Sibirica and C. recutita L. were 50% effective.

The results of the study allow us to conclude that there is a direct correlation between the volume of the preparation for an antibacterial effect and an inverse correlation between the dose of microbes for an antibacterial effect.

In the following experiment, we conducted studies to determine the antibacterial effect of the essential oils of the test preparation A. lerchiana and other herbal preparations widely used in veterinary medicine. Septarius was used as a test standard.

To determine the antibacterial effect in vitro, the method of disk diffusion in MPB was used.

Cultures of S. aureus, Streptococcus, and E. coli in doses of 500 million and 2 billion CFU were used as test microbes.

In the first part of the experiment, we studied the antibacterial effect of A. lerchiana in various doses (100%, 50%, 25%, 12.5%, 6.25%) against S. aureus, Streptococcus, and E. coli at a dose of 500 million CFU (Figure 1).

Figure 1
Diagram of the antibacterial effect of A. lerchiana in various doses against the test microbes.

The results of the experiment showed that the high concentration of A. lerchiana had a strong effect on all microbe cultures, especially on S. aureus.

Reducing the concentration to 50 and 25% reduced the antibacterial effect from 20 to 13-17 mm (by 35-50%) for Staphylococcus and Streptococcus. At concentrations of 12.5 and 6.25%, the antibacterial effect was determined as 30-40% against Staphylococcus and Streptococcus.

A decrease in the concentration of A. lerchiana essential oil significantly reduced the antibacterial effect against E. coli (at 50% concentration to 45%, at 25% concentration to 30%), while at a concentration of 12.5% and below, no antibacterial effect was observed.

Thus, the antibacterial effect of A. lerchiana against Staphylococcus and Streptococcus was noticeable even with a decrease in its concentration. Whereas the effect of A. lerchiana on E. coli was lower. We noted a direct correlation between its antibacterial effect against all microbe cultures.

In the second part of the experiment, we conducted a comparative analysis of the antibacterial effect of A. lerchiana compared to other plant-based preparations against the three types of microbial cultures. Septarius was used as a test antiseptic (Figure 2).

Figure 2
Diagram of the antibacterial effect of essential oils obtained from various plants against the test microbes in diffuse agar.

According to the results of the study, we observed a high antibacterial effect of A. lerchiana against Staphylococcus (24 mm at 500 million CFU, 20 mm at 1 billion CFU). Septarius showed an effect against Staphylococcus equaling 17 mm at 500 million CFU and 14 mm at 1 billion CFU. The remaining preparations showed an effect level of less than 13 mm.

Studies of the effect of essential oils on Streptococcus showed that A. lerchiana has the highest effect (19 mm at 500 million CFU and 15 mm at 1 billion CFU). Septarius has a relatively high effect (16 mm at 500 million CFU and 13 mm at 1 billion CFU). The remaining preparations were below the level of 12 mm.

A low antibacterial effect was observed in all preparations against E. coli. A. lerchiana ranged from 20 to 13 mm, and M. alternifolia and Septarius from 15 mm to 11 mm. The remaining preparations were below the level of 3 mm.

Thus, according to the results of experiments, we have noted the high antibacterial effect of A. lerchiana against Staphylococcus and Streptococcus, while the essential oils of other medicinal plants showed a low antibacterial effect. As for E. coli, the antibacterial effect of A. lerchiana, A. Sibirica, M. alternifolia essential oils and Septarius was higher than that of other medicinal plants (above 11 mm).

4. Discussion

The antimicrobial effect of different Artemisia species varies depending on the microorganisms (S. aureus, S. epidermidis, E. coli, C. albicans) and their effect on the ability of microorganisms to form biofilms. According to the study data, a bactericidal effect was detected in 14 Artemisia species used in the studies. Nine Artemisia species demonstrated bactericidal properties against S. epidermidis, six Artemisia species against S. aureus, and 11 Artemisia species against E. coli (Kartashova et al., 2009KARTASHOVA, O.L., UTKINA, T.M., ZHESTKOV, A.V., KURKIN, V.A. and ZOLOTAREV, P.N., 2009. Influence of phytosubstances with antioxidant activity on the persistent properties of microorganisms. Antibiotiki i Khimioterapiya, vol. 54, no. 9-10, pp. 16-18.). The results of our study showed that A. lerchiana had a high antibacterial effect against gram-positive microbes (Staphylococcus and Streptococcus). A. lerchiana was also highly effective against gram-negative E. coli but at a low dose of the test microbe (500 million CFU). With an increase in the dose of microbes, the growth rate of the microbes will increase to a high (76-100%), which is associated with a low antibacterial effect. After evaluating the antibacterial effect of A. lerchiana in comparison with Septarius, we note that it is not less effective than the test antiseptic against gram-positive microbes (Staphylococcus and Streptococcus). The effect is high for small doses of test microbes and average for large doses of test microbes (2 billion CFU).

A comparative analysis of A. lerchiana infusion with other infusions of the studied medicinal plants showed its superiority by 25% concerning Staphylococcus, Streptococcus, and E. coli. Only A. absinthium corresponded to the effect level of A. lerchiana at low doses of the test microbe (500 million CFU). Considering that the concentration of antibacterial active substances is higher in essential oils of plants than in their infusions, we note a more pronounced effect of the studied preparations on test microbes in the form of essential oils. Tsedenova et al. (1999)TSEDENOVA, L.P., ROMANOV, O.E., ROMANOVA, T.O. and LAVRENTIEVA, E.P., 1999. Antimicrobial activity of Artemisia lerchiana Web. ex Stechm., growing in Kalmykia. Rastitel’nye Resursy, vol. 35, no. 4, pp. 58-60. in their studies also show that some components of essential oil (1,8-cineol, borneol, α-thujone, bornyl acetate, α-thujone) obtained from A. lerchiana protect the body from pathogens and have an active fungicidal and bactericidal effect.

Studies of the antibacterial effect of A. lerchiana essential oil, depending on its concentration, have shown its effectiveness against Staphylococcus and Streptococcus even in low concentrations. However, the antibacterial effect against E. coli has been observed only at high concentrations. At low concentrations, there is no lysis zone. The results of our study are confirmed by the studies of other scientists from different countries and different climatic zones (Salimov et al., 2019SALIMOV, A.M., SHAROPOV, F.S., SANGOV, Z.G., MIRZOEV, S.O., SALIMOV, T.M. and KHAIDAROV, K.K., 2019. Antimicrobial properties of the essential oil of Artemisia annua growing in Tajikistan. Doklady Akademii Nauk Respubliki Tadzhikistan, vol. 62, no. 9-10, pp. 572-575.).

Comparative analysis of A. lerchiana with other medicinal plants showed its high antibacterial efficacy against all test microbes. The preparations closest to the effect level of A. lerchiana were A. Sibirica and M. alternifolia against Staphylococcus and Streptococcus, while the rest of the tested preparations were ineffective against all test microbes. In the evaluation of A. lerchiana, its antibacterial effects can be considered high, while the rest of the preparations had an average evaluation result. In other studies, an extract of A. lerchiana based on methanol and acetone also showed a bacteriostatic effect against Russula aeruginosa 1390, as well as against two other strains (E. coli K16, Bacillus subtilis 168) (Sotirova et al., 2022SOTIROVA, A., MUTAFOVA, B., BERKOV, S. and NIKOLOVA, M., 2022. Antibacterial activity of methanol extract and acetone exudates from Bulgarian plants. Acta Microbiologica Bulgarica, vol. 38, no. 1, pp. 48-51.).

We observed a direct dependence of the antibacterial effect of the preparation on its volume and the dose of test microbes. That is, the higher the volume of the preparation, the more effective the action, and the higher the dose of the test microbe, the more the antibacterial effect decreases. Even in small volumes and doses, A. lerchiana has a good antibacterial effect.

The obtained results of the study will be useful for scientists researching various medicinal products of plant origin and for veterinary specialists looking for therapeutic and preventive measures.

5. Conclusion

The results of the study show that A. lerchiana has a high antibacterial effect against gram-positive test microbes (Staphylococcus and Streptococcus). The most effective preparation is A. lerchiana in the form of essential oil when even its low concentration has an antibacterial effect.

A comparative assessment of A. lerchiana with Septarius shows that A. lerchiana has a similar effect in general and in the form of essential oil even surpasses the effect of Septarius. A. lerchiana surpasses medicinal plants used in the experiment in terms of antibacterial effect.

Thus, the results of the study allow us to recommend A. lerchiana to be used in production as an effective antibacterial agent. The study was limited to the comparative analysis of antibacterial effect of Artemisia lerchiana. Further research should be directed to the practical study of the effect of Artemisia lerchiana in the medical and veterinary activities.

Acknowledgments

This research was funded by the Science Committee of the Ministry of Education and Science of the Republic of Kazakhstan (Grant № AP15473422 «Determination of efficacy of Artemisia lerchiana based medication in the treatment of surgical injuries in animals»).

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Publication Dates

  • Publication in this collection
    10 Nov 2023
  • Date of issue
    2023

History

  • Received
    16 Aug 2023
  • Accepted
    11 Oct 2023
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