Antifungal and Antibacterial Activity of Aqueous and Ethanolic Extracts of Different Rosa rugosa Parts

Abstract

With the rising incidence of drug-resistant pathogens, focus should be placed on biologically active compounds derived from plant species used in herbal medicine, as these compounds may provide a new source of antifungal and antibacterial activities. The aim of this study was to evaluate the antifungal and antibacterial activity of ethanol and aqueous extracts from different parts of Rosa rugosa. In order to evaluate the antimicrobial activity of the extracts of R. rugosa rose hips, flowers, petals, leaves, stems, and roots, a laboratory microbiological test was performed using the well diffusion method in agar. A rotary evaporator was used for extract concentration and extractant removal. Antimicrobial activity was tested against one fungus, four Gram-positive, and four Gram-negative bacteria. The leaf extracts exhibited the strongest antimicrobial activity, followed by the extracts from the petals and rose hips, while weaker activity was observed in the root extracts. The extracts from the stems and rose hips showed the weakest effect. Ethanol extracts were more effective than water extracts. Aqueous and ethanolic extracts of R. rugosa parts demonstrated antifungal activity against Candida albicans, with ethanol extracts proving to be more effective. Among all the R. rugosa parts analyzed, the petals exhibited the strongest antifungal activity.

1. Introduction

Research on the antimicrobial properties of plants is important due to their potential applications in medicine, cosmetics, food, and agriculture, as they may help to develop natural alternatives to chemical preservatives, pesticides, and antibiotics. Additionally, plants are being evaluated as a potential new source of antifungal and antibacterial agents [1,2].

The literature increasingly mentions Rugosa rose (Rosa rugosa Thunb., Japanese rose) as a possible source of natural antioxidants, vitamins, flavonoids, and other bioactive substances that can have a positive effect on human health [3]. Rosa rugosa is a perennial shrub belonging to the family Rosaceae, the genus Rosa. It is most common in coastal areas—dunes and rocky coasts—but it is well adapted to grow in sandy, gravelly habitats of the mainland, so it is often found on slopes, roadsides, meadows, and forest edges [4,5,6]. Rosa rugosa is naturally distributed in East Asia, especially in China, Korea, Japan, and the Far East [5,6]. This plant has naturalized in various parts of the world, including North America, Canada, New Zealand, and much of Europe, particularly in northern, western, and central Europe [5,7]. Rosa rugosa was introduced to Europe from East Asia as an ornamental plant in the 19th century and is now considered an invasive species due to its rapid spread [5,8]. Although the plant is invasive, studies have shown that various parts of the R. rugosa contain many biologically active components, such as flavonoids, antioxidants, vitamins (especially vitamin C), essential oils, tannins, and organic acids, which have a positive effect on health and are used in both traditional and modern medicine. Research has demonstrated the antioxidant, anti-inflammatory, anticancer, antidiabetic, antifungal, and antibacterial properties of R. rugosa [3,9,10,11,12,13].

The antimicrobial properties of plants are determined by their phenolic and sulfur-containing compounds, as well as terpenoids and alkaloids [14]. Rosa rugosa is a natural source of phenolic compounds and terpenoids, contributing to its antimicrobial effect [15]. The mechanisms of antimicrobial action, which are characterized by the substances of plant origin, can be different. These include the disruption of bacterial cell wall synthesis or its structural integrity, damage to the cytoplasmic membrane, and the inhibition of DNA, RNA, proteins, or enzyme production essential for microbial survival [14,16]. Phytochemical compounds can have an antimicrobial effect on microbes only in one of the listed ways or as a combination of them, but all of them lead to the destruction of microorganisms [16]. The most widely analyzed part of R. rugosa in terms of antimicrobial properties is the rose fruits (rose hips). Recently, Cendrowski et al. [16] investigated the antimicrobial properties of R. rugosa against ten standard microorganism cultures by examining aqueous and ethanolic extracts concentrated by a rotary evaporator. It was found that the highest sensitivity was against Bacillus cereus, Escherichia coli, and Klebsiella pneumoniae bacteria. Previously, Olech et al. [10] analyzed the different parts of R. rugosa, flowers, leaves, roots, fruits, and seeds. Ethanol and aqueous were used as extractants. The effect was determined against four Gram-positive (Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus) and four Gram-negative (E. coli, K. pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis) bacterial cultures. Leaves showed the strongest antibacterial properties, followed by flowers and roots, while rose fruits and seeds had the weakest activity. The results obtained during this study allowed us to assume the potential benefit of the R. rugosa in the antiseptic effect on the mucous membrane of the upper respiratory tract and the digestive tract [10].

Few studies have investigated the antifungal effects of R. rugosa plant parts. Previously, Nowak et al. [11] evaluated the antifungal activity of R. rugosa petals collected in Poland against Candida albicans, Candida parapsilosis, and various bacterial strains. Rosa rugosa petals were found to possess significant antifungal and antibacterial properties. Additionally, extracts obtained from rose fruits and leaves have demonstrated activity against Candida spp. [17,18].

The R. rugosa rose hips and flowers are already widely used both in traditional medicine and in the food industry, and a number of scientific studies have been conducted on these parts of the plant. However, to better understand the potential of the whole plant, it is necessary to focus more research on its other parts, such as stems and leaves, as these parts also exhibit significant antimicrobial activity. The bioactive properties of these plant parts could widen the range of natural antifungal and antibacterial agents and provide additional opportunities for their application [19]. In Lithuania, no comprehensive and wide-scale studies have been conducted on the antimicrobial activity of different parts of R. rugosa. However, further studies of the parts of R. rugosa may reveal new possibilities of the application of this plant.

The aim of this study was to evaluate the antifungal and antibacterial activity of ethanol and aqueous extracts from different parts of Rosa rugosa.

2. Materials and Methods

2.1. Sample Collection

The different parts of the R. rugosa were collected from natural habitats in the Neringa municipality located in the Curonian Spit in Lithuania (Figure 1). Only visually healthy plants growing in natural habitats were collected. The collection time and drying of plant raw materials were carried out based on the recommendations of Olech et al. [10].

Rosa rugosa rose hips, flowers, petals, leaves, stems, and roots were collected from the same wrinkled-leaf rose plants but at different periods corresponding to the formation of specific plant parts. The flowers, petals, and leaves were collected in May 2023, when the plant began to bloom. The rose hips were collected in August 2023, when they ripened. The stems and roots were collected in the autumn of the same year.

The collected plant material was dried in a thin layer spread on paper at room temperature in a place protected from direct sunlight. The dried plant parts were stored in dark, airtight jars until the time of the study.

2.2. Extraction Preparation

To increase the surface area of the plant’s raw material, the dried plant parts were ground. Ethanol and aqueous extracts were prepared according to Olech et al. [10]. In total, 10 g of dried and ground parts of each R. rugosa were weighed to prepare ethanolic extracts, which were infused with 50 g of 70% ethanol and stored for 7 days in sealed glass vials with ground stoppers, with occasional stirring. After 7 days, the obtained extracts were filtered and filled to 70% ethanol to the original volume before filtration. Aqueous extracts were prepared by pouring 2 g of dried and ground parts of each plant into 100 mL of boiling water, leaving them to cool, and filtering and filling with water to the original volume before filtration.

2.3. Removal of Extractants

A rotary evaporator was used for the removal of the solvents (i.e., ethanol and water) and the concentration of the extracts. The filtered extract was poured into a round-bottomed flask. The water bath temperature was 60 °C, and the temperature of the water flowing through the condenser was 20 °C. The vacuum was selected based on the requirements to boil at 40 °C; a vacuum of 175 millibars was required for ethanol, while a vacuum of 72 millibars was needed for water [16]. The flask rotation speed used was 200 rotations per minute. The process can be most effectively accelerated by increasing the vacuum, the flask’s rotation speed, or raising the water bath temperature; however, the latter should be avoided, as it may cause the degradation of the thermolabile components in the extract [20,21]. The process was stopped when the solvent in the round-bottomed flask evaporated, and no more condensate dripped into the waste flask.

2.4. Concentration Calculation

To determine the mass of the concentrated extract formed on the walls of the round-bottomed flask, the flask was weighed after evaporation, and the mass of the empty flask, which was weighed before the experiment, was subtracted. The resulting concentrated extract was then dissolved in a volume of 10% dimethylsulfoxide (DMSO) to obtain a final test extract concentration of 100 mg/mL. DMSO is the most commonly used solvent in such studies by researchers.

2.5. Determination of Antimicrobial Activity

The antimicrobial activity of the wrinkled-leaf rose parts extracts was determined using the agar well diffusion method. For this, Mueller–Hinton agar was prepared according to the manufacturer’s instructions, boiled seven times to its boiling point, and poured into Petri dishes, with 40 mL of medium in each. Once the agar solidified, 6.00 mm diameter wells were made using glass Pasteur pipettes. The agar plates with the wells were inoculated with 0.5 McFarland turbidity microbial suspensions, which were prepared by emulsifying the colonies of the microorganisms to be tested in 2 mL of physiological saline. Antimicrobial activity was tested against Gram-positive bacteria: Bacillus cereus (ATCC 11778), Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 12228), Enterococcus faecalis (ATCC 29212), Gram-negative bacteria: Proteus mirabilis (ATCC 29906), Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (ATCC 27853), and Escherichia coli (ATCC 25922), and one fungus (Candida albicans (ATCC 10231)).

After inoculating the microorganism suspensions, 100 μL of the test extracts were added to the wells and incubated in a thermostat for 24 h at 35–37 °C. After incubation, the formation of inhibition zones and the nature of the antimicrobial activity (microbicidal/microbiostatic) were assessed. The diameter of the inhibition zones, including the diameter of the well itself, was measured with a ruler and recorded in millimeters (mm). If a clear zone around the well filled with the extract was observed, where no microbial colonies were visible, the antimicrobial activity was considered microbicidal. If only the inhibition of microbial growth was observed in the zone, the activity was classified as microbiostatic. If the inhibition zone around the well did not exceed 6 mm, the test extract was considered to have no antimicrobial activity. To practically verify that the solvent used in the experiment had no antimicrobial properties, a negative solvent control was performed, using a well filled with 100 μL of 10% DMSO. If no inhibition zone formed around the well after incubation, it was concluded that the solvent had no antimicrobial activity. To ensure the accuracy of the results, the experiments were repeated three times, and the arithmetic averages of the obtained results were calculated.

2.6. Statistical Data Analysis

ANOVA and Tukey’s Honest Significant Difference (HSD) test was used to evaluate the reliability of the differences in the mean of the inhibitory zones, and the significance of the pairwise differences between the group means was calculated. The difference between group means was considered statistically significant when p < 0.05.

3. Results

According to the data on the antifungal and antibacterial activity of the various aqueous and ethanolic extracts of the different parts of R. rugosa for microorganisms, it was determined that ethanolic and aqueous extracts of all the tested parts of R. rugosa (rose hips, petals, flowers, leaves, stems, and roots) showed antifungal, antimicrobial activity against Gram-positive and Gram-negative microorganisms and only their spectrum and strength differed.

The antifungal activity of the aqueous and ethanolic extracts of the R. rugosa parts against C. albicans was evaluated by measuring the average diameter of the inhibition zones (mm) across three replicates. The results demonstrated that the aqueous and ethanolic extracts of the R. rugosa parts exhibited antifungal activity against C. albicans. Ethanol extracts were more effective against C. albicans. Of all the analyzed parts of R. rugosa, the petals showed the strongest antifungal activity (Figure 2).

The antimicrobial activity of R. rugosa hips ethanolic and aqueous extracts against Gram-positive B. cereus, S. aureus, and S. epidermidis and Gram-negative E. coli was determined. Neither the aqueous nor the ethanolic extracts of these rose hips had antimicrobial activity against E. faecalis, P. mirabilis, and K. pneumoniae (Figure 3).

The aqueous and ethanolic extracts of R. rugosa flowers also exhibited antimicrobial activity against all tested Gram-positive microorganisms, B. cereus, S. aureus, S. epidermidis, and E. faecalis and Gram-negative P. mirabilis, P. aeruginosa, E. coli, except Gram-negative K. pneumoniae, which was affected by the ethanolic extract, while the aqueous extract had no antimicrobial effect (Figure 4).

The antimicrobial activity of the R. rugosa petals’ ethanolic extracts against all Gram-positive B. cereus, S. aureus, S. epidermidis, and E. faecalis and all Gram-negative P. mirabilis, K. pneumoniae, P. aeruginosa, and E. coli was determined. All the aqueous extracts of the R. rugosa petals acted against Gram-positive and Gram-negative except Gram-positive E. faecalis and Gram-negative K. pneumoniae (Figure 5).

According to the obtained data, the strongest antimicrobial activity was shown by the leaves of R. rugosa. The weakest action was observed against the Gram-negative bacterium E. coli, and the strongest activity was determined against the Gram-positive bacterium S. epidermidis (Figure 6).

The spectrum of the antimicrobial activity of roots was the narrowest, showing antimicrobial activity against the least studied types of microorganisms. The antimicrobial activity of R. rugosa roots’ ethanolic and aqueous extracts against Gram-positive B. cereus, S. aureus, and S. epidermidis and Gram-negative E. coli was determined. Ethanolic extracts of R. rugosa roots acted against Gram-negative P. aeruginosa and E. coli. Neither aqueous nor ethanolic extracts of R. rugosa roots had antimicrobial activity against E. faecalis, P. mirabilis, and K. pneumoniae (Figure 7).

The antimicrobial activity of R. rugosa stems’ ethanolic and aqueous extracts against Gram-positive B. cereus, S. aureus, S. epidermidis, and E. faecalis and Gram-negative E. coli was determined. The ethanolic extracts of R. rugosa stems acted against Gram-negative P. aeruginosa, E. coli. Neither aqueous nor ethanolic extracts of stems had antimicrobial activity against P. mirabilis and K. pneumoniae (Figure 8).

According to the results, ethanolic and aqueous extracts from all tested parts of R.rugosa (rose hips, petals, flowers, leaves, stems, and roots) exhibited antifungal activity. The highest average inhibition zones were observed for the ethanolic extracts of petals (20.00 mm) and flowers (17.00 mm), and their activity was fungistatic. In contrast, the ethanolic and aqueous extracts of leaves demonstrated fungicidal activity, with average inhibition zone diameters of 17.00 mm and 14.00 mm, respectively. The extracts from other plant parts exhibited fungistatic activity only, with mean inhibition zone diameters ranging from 11.33 mm for the aqueous rose hip extract to 20.00 mm for the ethanolic petal extract (

Table 1. Comparison of antimicrobial activity of different parts of Rosa rugosa aqueous and ethanolic extracts’ microorganisms expressed as the three attempts average diameter of inhibition zones, mm.

	Average Diameter of Inhibition Zones, mm
	Plant Rosa rugosa Parts
	Leaves		Petals		Flowers		Roots		Stems		Rose Hips
Type of Extract Microorganism	E	A	E	A	E	A	E	A	E	A	E	A
C. albicans	17.00 ± 1.00	14.00 ± 1.00	20.00 ± 1.00	14.00 ± 1.00	17.00 ± 1.00	14.00 ± 1.00	15.00 ± 1.00 ‘	12.33 ± 0.58	14.67 ± 1.15 ‘	12.00 ± 1.00	14.33 ± 0.58 ‘	11.33 ± 0.58 “
B. cereus	19.67 ± 1.15	15.67 ± 1.15	19.33 ± 0.58	15.67 ± 1.15	17.67 ± 1.15 *	15.33 ± 0.58	13.00 ± 1.00 *	9.00 ± 1.00 **	13.00 ± 1.00 *	12.33 ± 0.58 **	12.67 ± 1.15 *	11.33 ± 0.58 **
S. aureus	19.33 ± 0.58	13.00 ± 1.00	17.33 ± 0.58 *	10.00 ± 1.00 **	16.67 ± 1.15 *	8.00 ± 1.00 **	12.00 ± 1.00 *	10.00 ± 1.00 **	12.00 ± 1.00 *	11.33 ± 0.58	11.00 ± 1.00 *	9.00 ± 1.00 **
S. epidermidis	22.67 ± 1.15	17.00 ± 1.00	24.67 ± 1.15	16.00 ± 1.00	20.00 ± 1.00	10.00 ± 1.00 **	11.67 * ± 1.15	10.00 ± 1.00 **	11.67 * ± 1.15	11.33 ± 0.58	10.67 * ± 1.15	10.00 ± 1.00
E. faecalis	14.00 ± 1.00	-	12.00 ± 1.00	-	12.33 ± 0.58	-	8.00 * ± 1.00	8.00 ± 1.00	-	-	-	-
E. coli	9.00 ± 1.00	19.33 ± 0.58	20.00 ± 1.00	19.00 ± 1.00	19.67 ± 1.15	18.00 ± 1.00	10.67 ± 1.15 ‘*	10.33 ± 0.58 “*	14.33 ± 0.58 ‘*	13.67 ± 1.15 “*	13.67 ± 1.15 ‘*	11.00 ± 1.00 “*
P. aeruginosa	21.33 ± 0.58	15.00 ± 1.00 “*	22.00 ± 1.00	9.00 ± 1.00	20.00 ± 1.00	8.00 ± 1.00	10.00 ± 1.00 ‘*	-	10.33 ± 0.58 ‘*	-	10.00 ± 1.00 ‘*	-
P. mirabilis	16.33 ± 0.58	15.33 ± 0.58	14.33 ± 0.58	13.00 ± 1.00	13.00 ± 1.00	9.33 ± 0.58 “*	-	-	-	-	-	-
K. pneumoniae	9.67 ± 1.15	-	15.00 ± 1.00	-	15.67 ± 1.15	-	-	-	-	-	-	-

* A—aqueous extract; E—ethanolic extract- Fungicidal activity, - fungistatic activity, - bactericidal activity, and - bacteriostatic activity, ‘ p < 0.05, a statistically significant difference in the averages of the inhibitory zones was determined by comparing the ethanolic extracts of various parts of R. rugosa with the ethanolic extract of the petals, which has a fungistatic effect, “ p < 0.05, a statistically significant difference in the averages of the inhibitory zones was determined when comparing the aqueous extracts of various parts of R. rugosa with the aqueous extract of the petals, which has a fungistatic effect. * p < 0.05, a statistically significant difference in the averages of the inhibitory zones was determined by comparing the ethanolic extracts of various parts of R. rugosa with the ethanolic leaf extract, which has a bactericidal effect, ** p < 0.05, a statistically significant difference in the averages of the inhibitory zones was determined when comparing the aqueous extracts of various parts of R. rugosa with the aqueous leaf extract, which has a bactericidal effect. ‘* p < 0.05, determined statistically significant difference in the averages of the inhibitory zones comparing the ethanolic extracts of various parts of R. rugosa with the ethanolic extract of the petals, which has a bacteriostatic effect, “* p < 0.05, a statistically significant difference in the averages of the inhibitory zones was determined when comparing the aqueous extracts of various parts of R. rugosa with the aqueous extract of the petals, which has a bacteriostatic effect.

).

Only the ethanolic extract from R. rugosa leaves had the highest bactericidal activity against all tested Gram-positive and Gram-negative microorganisms. Both the ethanolic and aqueous extracts of R. rugosa parts (rose hips, petals, flowers, leaves, stems and roots) were active against Gram-positive bacteria B. cereus, S. aureus, and S. epidermidis. Also, the ethanolic extract of rose hips, petals, flowers, stems, roots and the aqueous extracts of petals, flowers, and roots had bactericidal activity against Gram-positive E. faecalis. All the Gram-negative bacterium P. mirabilis, K. pneumoniae, P. aeruginosa, and E. coli was sensitive to the ethanolic and aqueous extracts made from all tested parts of R. rugosa: rose hips, petals, flowers, leaves, stems, and roots. But the only ethanol extract of the leaves showed bactericidal activity against these microorganisms. In other cases, only bacteriostatic activity was detected, or the extracts did not work.

4. Discussion

In our study, the antifungal and antimicrobial activities of extracts from different parts of R. rugosa were analyzed in detail for the first time in Lithuania. To evaluate the antifungal and antibacterial effects of the different parts of R. rugosa, the activity of their ethanolic and aqueous extracts was analyzed against C. albicans and eight bacterial species: B. cereus, S. aureus, S. epidermidis, E. faecalis, P. mirabilis, K. pneumoniae, P. aeruginosa, and E. coli. Our study’s results showed that the ethanolic and aqueous extracts of all the studied parts of R. rugosa (rose hips, petals, flowers, leaves, stems, and roots) showed antimicrobial activity. The lowest activity was observed against the Gram-negative bacterium E. coli, and the strongest was found against the Gram-positive bacterium S. epidermidis. The data revealed that the leaves of R. rugosa exhibited the strongest antimicrobial activity. Among all the tested extracts, the ethanolic extracts from the leaves were the only ones to demonstrate a microbiocidal effect against all the tested microorganisms, including C. albicans, K. pneumoniae, P. mirabilis, P. aeruginosa, E. coli, E. faecalis, S. epidermidis, S. aureus, and B. cereus. In a previous study, the methanolic extracts of 37 plants and R. rugosa leaves were analyzed for S. mutans and C. albicans. Rosa rugosa leaves were characterized by moderate antimicrobial activity—the growth inhibition of a liquid medium was about 50%. The other part of the study was also carried out using the disk diffusion method—a concentration of 2 mg of extract/disk led to the formation of an inhibition zone of 12 mm against C. albicans and 10 mm against S. mutans. This suggests that R. rugosa leaves have the potential to combat oral microorganisms [18,22,23]. Wu et al. [24] analyzed waste extracts of Rosa setate and R. rugosa plant parts and found that after purification, the extracts have more effective bacteriostatic effects against bacteria (S. aureus, E. coli, S. epidermidis, P. aeruginosa) and fungus (C. albicans) than crude extracts. Ren et al. [25] analyzed the antimicrobial properties of R. rugosa flowers. This study did not use extracts, but fluid extracted from the central vacuoles of the flower cells. The minimum inhibition concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) were measured, and the obtained results revealed that the flowers had the best effect against Fusobacterium nucleatum, Propionibacterium acnes, and S. aureus bacteria, and the effect against the fungus C. albicans was the weakest. In our study, the ethanolic extracts of petals and flowers also exhibited antimicrobial effects against all tested organisms, but their activity was weaker. Only Gram-positive bacteria had microbiocidal activity, and only microbiostatic effects were detected against Gram-negative bacteria and the C. albicans fungus. Kamijo et al. [26] investigated the effect of dried R. rugosa petal powder against intestinal bacteria—their test showed that R. rugosa petals reduced the growth of pathogenic bacteria (Bacteroides vulgatus, E. coli, S. aureus, B. cereus). In our study, the ethanolic extracts of the R. rugosa roots also had a microbiocidal effect on Gram-positive microorganisms. Their spectrum of action against Gram-negative bacteria was narrower—they were completely inactive on P. mirabilis and K. pneumoniae bacteria, while the action of C. albicans against the root extracts was fungistatic. The antimicrobial properties of R. rugosa roots were investigated by Olech et al. [27], and according to their data, R. rugosa roots have moderate antimicrobial activity against S. epidermidis, S. aureus, B. subtilis, M. luteus, E. coli, K. pneumoniae, P. aeruginosa, P. mirabilis, C. albicans, and C. parapsilosis.

In our study, the spectrum of the antimicrobial activity of the R. rugosa stems and rose hips was the narrowest, showing antimicrobial activity against the least studied types of microorganisms. Both aqueous and ethanolic extracts of these plant parts showed no antimicrobial activity against E. faecalis, P. mirabilis, and K. pneumoniae bacteria. The best antimicrobial activity of the leaves, petals, flowers, and roots of R. rugosa can be attributed to the abundance of phenolic compounds found in these parts of the plant. Olech et al. [10] found a correlation between the number of phenolic compounds found in the parts of R. rugosa and its antimicrobial properties. The parts with a higher number of phenolic compounds had higher antimicrobial activity and it was found that leaves, flowers, and roots had the strongest antibacterial properties, while rose hips and seeds had the weakest activity. Our study also found that the R. rugosa antimicrobial activity of leaf extracts was the strongest, and petal, flower, and root extracts were weaker, and stem and rose hip extracts were the weakest. According to the study results, the ethanolic extracts of all the tested parts of R. rugosa exhibited stronger antimicrobial effects than the aqueous extracts. The only exception was the root extracts against E. faecalis, where both ethanolic and aqueous extracts showed microbiocidal effects of equal strength, with average inhibition zone diameters of 8.00 mm. How different extractants are able to extract the active substances from the plant is determined by the different polarity of the extractants [28]. Therefore, when determining the antimicrobial properties of a plant, the choice of extractant to be used in the study has a significant influence. When comparing the antimicrobial effect of the extracts of parts of R. rugosa on microorganisms of different characteristics, it was found that the most sensitive to the tested extracts were Gram-positive bacteria B. cereus and S. aureus. Data on the strong antibacterial action of R. rugosa against these microorganisms were also found in a recent study. Cendrowski et al. [16] reported that among the tested Gram-positive bacteria, B. cereus exhibited the highest sensitivity to R. rugosa fruit extracts. Among Gram-negative bacteria, E. coli and K. pneumoniae showed the greatest sensitivity, while E. faecalis and P. mirabilis cultures were the most resistant to the extracts. According to the researchers, the aqueous extracts of R. rugosa fruits were more effective than the ethanol extracts. The obtained data allowed us to assume that R. rugosa fruits could be a potential food additive, reducing their bacterial contamination. Olech et al. [17] used methanol for the extraction of the R. rugosa rose hips, and they were treated with the resulting extract of eight cultures of bacteria and two fungi of the genus Candida. The authors found moderate antimicrobial activity against both bacteria (MIC 0.625–2.5 mg/mL) and fungi (MIC 1.25 mg/mL). Although all tested extracts also had an antimicrobial effect against the Gram-positive bacterium S. epidermidis, the nature of the action was not bactericidal in all cases—the aqueous extracts of the stems and rose hips showed only bacteriostatic action. Results on the strong potential of R. rugosa against various staphylococcal bacteria were also reported by Milala et al. [29]. Milala et al. [29] compared the action of the rose hips of three thorn species (R. rugosa, Rosa canina, Rosa pomifera) against staphylococcal bacteria. Reference cultures and pathogens isolated from food were studied. According to the data of this test, it was found that the R. rugosa species had the strongest anti-staphylococcal effect and that the entire rose hip had better antimicrobial properties than only its soft part. Among the Gram-positive bacteria examined in this study, E. faecalis showed the highest resistance to the tested extracts—extracts made from rose hips and stems had no effect against it, while only the ethanolic extracts of leaves, petals, and flowers were effective. Only the roots differed in the equal bactericidal effect of both ethanolic and aqueous extracts against this bacterium. The results of the study showed that only the ethanolic extract prepared from the leaves had a bactericidal effect on all tested Gram-negative bacteria.

Among Gram-negative bacteria, E. coli was the most sensitive to the tested extracts, against which all the analyzed extracts had an antimicrobial effect. The ethanolic extract of the leaves caused bactericidal, and the remaining extracts caused bacteriostatic antimicrobial effects. P. aeruginosa was less sensitive to the extracts—compared to E. coli, it showed resistance to aqueous extracts made from rose hips, stems, and roots. According to our study, P. mirabilis was sensitive only to the ethanolic and aqueous extracts of leaves, petals, and flowers, while extracts from rose hips, stems, and roots did not exhibit antimicrobial activity against this bacterium. The bacterium K. pneumoniae had the lowest antimicrobial activity among the extracts tested in this study. Klebsiella pneumoniae was sensitive only to the ethanolic extracts of leaves, petals, and flowers. Leaf extracts showed microbicidal activity, while petals and flowers showed microbicidal activity. It can be assumed that the resistance of K. pneumoniae bacteria can be determined by its structural features, since it is a bacterium that forms a polysaccharide capsule. Analyzing the antifungal activity of C. albicans, it was found that the fungus is killed by ethanolic and aqueous extracts prepared from the leaves of R. rugosa. The extracts of other parts of this plant tested showed only a growth-inhibiting effect. In the conducted study, there was a tendency that the tested extracts had the strongest antimicrobial properties against Gram-positive bacteria, the antifungal activity was weaker, and the least sensitive to the tested extracts were Gram-negative bacteria. Given that the antimicrobial activity of plant-derived substances is mostly due to the phenolic compounds found in them, the action of which is directed at causing damage to the wall and membrane of the microorganism, such results can be attributed to the structural differences in the studied microorganisms [16,30,31].

5. Conclusions

The data of this study showed that all tested extracts had antimicrobial properties, and they differed only in their potency and spectrum of action. The best efficiency was shown by the leaves of the R. rugosa, followed by the petals and flowers, and the roots showed a weaker effect, and the effects of the stems and thistle extracts were the weakest. A tendency was observed that ethanolic extracts worked more efficiently than aqueous ones. The most sensitive to the tested extracts were Gram-positive bacteria, and the antifungal activity was weaker, and the group of Gram-negative bacteria showed the highest resistance. The tested extracts had the most effective effect on the bacteria B. cereus and S. aureus, and the antifungal activity was weaker. All the tested extracts had a bactericidal effect against them, while the most resistant to the tested extracts were P. mirabilis and K. pneumoniae bacteria.