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Kernou, O.; Azzouz, Z.; Madani, K.; Rijo, P. Antimicrobial and Antibiofilm Activities of Rosmarinic Acid. Encyclopedia. Available online: https://encyclopedia.pub/entry/45557 (accessed on 12 May 2024).
Kernou O, Azzouz Z, Madani K, Rijo P. Antimicrobial and Antibiofilm Activities of Rosmarinic Acid. Encyclopedia. Available at: https://encyclopedia.pub/entry/45557. Accessed May 12, 2024.
Kernou, Ourdia-Nouara, Zahra Azzouz, Khodir Madani, Patrícia Rijo. "Antimicrobial and Antibiofilm Activities of Rosmarinic Acid" Encyclopedia, https://encyclopedia.pub/entry/45557 (accessed May 12, 2024).
Kernou, O., Azzouz, Z., Madani, K., & Rijo, P. (2023, June 14). Antimicrobial and Antibiofilm Activities of Rosmarinic Acid. In Encyclopedia. https://encyclopedia.pub/entry/45557
Kernou, Ourdia-Nouara, et al. "Antimicrobial and Antibiofilm Activities of Rosmarinic Acid." Encyclopedia. Web. 14 June, 2023.
Antimicrobial and Antibiofilm Activities of Rosmarinic Acid
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This entry is adapted from the peer-reviewed paper 10.3390/molecules28104243

Rosmarin is an original plant compound listed among the hydroxycinnamic acids. This substance has been widely used to fight microbial pathology and chronic infections from microorganisms like bacteria, fungi and viruses. Also, various derivatives of rosmarinic acid, such as the propyl ester of rosmarinic acid, rosmarinic acid methyl ester or the hexyl ester of rosmarinic acid, have been synthesized chemically, which have been isolated as natural antimicrobial agents. Rosmarinic acid and its derivatives were combined with antibiotics to obtain a synergistic effect. 

rosmarinic acid antimicrobial resistance synergistic effect antibiofilm

1. Introduction

Antimicrobial resistance (AMR) is a significant public health problem. According to ECDC and WHO estimates, more than 670,000 illnesses per year in the European area are caused by antibiotic-resistant bacteria, and over 33,000 human deaths are directly caused by these infections [1]. Antimicrobial resistance is directly related to the indiscriminate use of antibiotics and is developing at extremely high rates worldwide, including African countries like Cameroon [2]. Despite inadequate laboratory capacity to monitor AMR, the African continent shows evidence of the global trend of increasing drug resistance.
Some germs can not only spread in hospitals, but also in the great outdoors, where a significant resistance has been observed [3]. With increased drug consumption, increased hospital admissions, and increases in deaths, the financial consequences of AMR are financially devastating, including astronomical medical costs [4].
Antibiotic-resistant pathogenic bacteria include P. aeruginosa, S. aureus, Enterobacteriaceae and Enterococcus spp., while infections caused by nonresistant germs are significantly more difficult to treat [5].
In addition, the current overuse of conventional antibiotics and their improper use have reduced their efficacy, creating a serious problem for world health as well as development, and as such, the antimicrobial drug range is rapidly expanding compared to the available therapeutic treatments, making the latter ineffective or even inefficient [6]. The demand for new antimicrobial products is high, and products derived from natural sources are seen as a promising solution, including plant polyphenols, such as rosmarinic acid, which is a natural substance with antimicrobial activity.

2. Antimicrobial Activity

There have been hundreds of research studies conducted on the antimicrobial activity of AR (Table 1). This section will mention the most recent studies conducted, specifically, RA is used as a natural phytogenic additive in animal and poultry nutrition to improve their overall health, performance measures, the digestive system’s structure and function and its potential to modify the intestinal microbiota and decrease the number of disease-causing bacteria such as Salmonella spp, E. coli, and several other species of harmful bacteria [7][8]. Rosemary extracts may contain RA as the primary bioactive antimicrobial agent. However, using the methanol extract containing about 30% carnosic acid, with 16% carnosol and 5% RA, Gram-positive and Gram-negative bacteria were shown to be sensitive to rosemary, making it an excellent antibacterial, in contrast to an aqueous extract containing 15% RA which had more limited effects [9].
Due to their intense antimicrobial properties, medicinal plants, herbs and their oils are attracting great interest as innovative and alternative drugs, such as RA [10].
According to Benedec et al. [11], RA showed better antioxidant activity in vitro (DPPH technique) as well as significant action towards the Gram-positive bacteria. In addition, Rosmarinus officinalis extract showed even greater inhibition of the growth of Gram-positive bacteria than the Gentamicin control (Candida albicans). On the other hand, these researchers noted that this extract was without effect toward Gram-negative bacteria such as S. typhimurium, L. monocytogenes, E. coli, S. aureus, and C. albicans were found to be resistant to RAs derived from: Hyssopus officinalis L., M. officinalis L., O. vulgare L. [11]. RA addition decreases the rate of mortality in Japanese encephalitis virus-infected mice. Compared to animals infected with no RA treatment, the viral load was greatly reduced (p < 0.001) in RA-treated infected rats 8–9 days after infection [12].
The antibacterial properties of tannic acid have long been recognized as effective against both methicillin-resistant Staphylococcus aureus and other microorganisms [13]. Currently, one of the molecules used as a target for antibacterial polymer applications is the tannic acid–polymer metal complex [14]. Hospitalized patients are severely harmed by S. aureus, and tannic acid is known to be an inhibitor of various resistance phenotypes of S. aureus [15]. Furthermore, by reducing cell counts and numbers, RA inhibits the development of S. carnosus LTH1502 and E. coli K-12 [16].
Moreno et al. [9] examined extracts of Rosmarinus officinalis through a combination of biological tests. Antimicrobial activities were analyzed by both disk diffusion and dilution broth techniques. Gram-positive bacteria, including S. aureus, B. megaterium, B. subtilis, and E. faecalis, were more sensitive to the methanolic extract, which contains 30% carnosic acid, 16% carnosol, and 5% rosmarinic acid (minimum inhibition concentration (MIC), 2 to 15 mg/mL). Gram-negative bacteria such as K. pneumoniae, E. coli, X. campestris pv. campestris, and P. mirabilis were also treated with MIC 2 to 60 mg/mL, as well as yeasts such as S. cerevisiae, C. albicans, and P. pastoris (MIC of 4 mg/mL). However, the aqueous extract with a 15% rosmarinic acid content only exhibited a narrow spectrum of activity. The MICs for methanolic and water extracts correlated significantly with the values for pure carnosic acid and rosmarinic acid. So, these results indicated a good performance in relation to the antimicrobial efficacy with rosemary extracts combined with the relevant phenolic extracts. The principal antimicrobial bioactive agents in rosemary extracts were suggested to be carnosic acid or rosmarinic acid. From the point of view of practicality, it could be considered as a good nutritional supplement and herbal pharmaceutical product.
Rosmarinic acid has antibacterial properties against Staphylococcus aureus, E. coli, B. subtilis, and Salmonella. Hayriye [17] tested the effect of natural phenolic compounds extracted from vegetables, fruits, herbs and spices against these pathogens and E. coli had minimum bactericidal concentrations (MBC) of 0.9 mg/mL and minimum inhibitory concentrations (MIC) of 0.8 mg/mL. Salmonella had MIC and MBC of 0.9 and 1.0 mg/mL, respectively. Staphylococcus aureus and B. subtilis had MIC and MBC values of 1.0 and 1.1 mg/mL [17], respectively.
The strains LM1, LM2, and LM3 of L. monocytogenes were analyzed, and the presence of rosmarinic acid was shown to have no antibacterial effect over the incubation period of 60 h [18]. Previously, rosmarinic acid has been shown to exhibit high susceptibility to Gram-negative bacteria when exposed to rosmarinic acid, after 60 h of incubation, Salmonella species showed substantial levels of antimicrobial resistance, and the MICs of rosmarinic acid for S. enteridis, S. choleraesuis subsp., and S. paratyphi were less than 20 ppm [18].
Furthermore, rosmarinic acid had previously been recognized as an anti-HIV drug capable of inhibiting HIV replication [19]. The discovery of nitro and dinitro-rosmarinic acids, which inhibit viral replication by blocking HIV-I integrase, has significantly enhanced the anti-HIV efficacy of rosmarinic acid [20].
Table 1. Rosmarinic acid and its derivatives are used as antibiotics against several pathogenic microorganisms.
Pathogenic Microorganisms Active Concentrations References Pathogenic Microorganisms Active Concentrations References
Staphylococcus epidermidis 5001
Stenotrophomonas maltophilia
Enterococcus faecalis C159-6
Staphylococcus lugdunensis T26A3
Pseudomonas aeruginosa ATCC 27583		 MIC (0.3 mg/mL of RA)
MIC (0.3 mg/mL of RA)
MIC (0.3 mg/mL of RA)
MIC (0.6 mg/mL of RA)
MIC (2.5 mg/mL of RA)		 [21] Escherichia coli MIC 0.8 mg/mL of RA; MBC 0.9 mg/mL of RA [17]
Staphylococcus aureus MIC 1.0 mg/mL of RA; MBC 1.1 mg/mL of RA
Salmonella MIC 0.9 mg/mL of RA; MBC 1.0 mg/mL of RA
Bacillus subtilis MIC 1.0 mg/mL of RA; MBC 1.1 mg/mL of RA
Corynebacterium T25-17
Mycobacterium smegmatis 5003
Staphylococcus warneri T12A12		 MIC (2.5 mg/mL of RA)
MIC (1.2 mg/mL of RA)
MIC (1.2 mg/mL of RA)		 Micrococcus luteus MIC 0.1 mg/mL; MBC 0.2 mg/mL [22]
Rothia mucilagenosa MIC 0.1 mg/mL; MBC 0.2 mg/mL
Klebsiella sp. IZ 28 mm at 1 mg/mL of RA [23] Streptococcus agalactiae MIC 0.05 mg/mL; MBC 0.1 mg/mL
Stenotrophomonas maltophela IZ 19 mm at 1 mg/mL of RA Streptococcus angiosus MIC 0.05 mg/mL; MBC 0.1 mg/mL
Streptomyces sp. IZ 26 mm at 1 mg/mL of RA Streptococcus dysgalactie MIC 0.05 mg/mL; MBC 0.1 mg/mL
Pantoea agglomerans IZ 18 mm at 1 mg/mL of RA Streptococcus oralis MIC 0.05 mg/mL; MBC 0.1 mg/mL
Paenibacillus chibensis IZ < 1 mm at RA-methyl ester
IZ 4.4 mm at tannic acid
IZ > 2 mm at RA-hexyl ester
IZ between 3 mm and 4 mm at RA-propyl ester		 [24] Streptococcus parasanquinis MIC 0.05 mg/mL; MBC 0.1 mg/mL
Streptococcus pyogenes MIC 0.1 mg/mL; MBC 0.2 mg/mL
Streptococcus salivarius MIC 0.002 mg/mL; MBC 0.004 mg/mL
Staphylococcus waeneri IZ < 1 mm at RA-methyl ester
IZ 5 mm at tannic acid
IZ ˃ 2 mm at RA-hexyl ester
IZ between 2 mm and 3 mm at RA-propyl ester		 Staphylococcus aureus MIC ˃ 0.8 mg/mL; MBC ˃ 0.8 mg/mL
Staphylococcus hominis MIC 0.4 mg/mL; MBC 0.8 mg/mL
Bacillus cereus IZ > 3 mm at RA-methyl ester
IZ 6 mm at tannic acid
IZ 7.7 mm at RA-hexyl ester
IZ 9 mm at RA-propyl ester		 Enterobacter cloacae MIC 0.1 mg/mL; MBC 0.2 mg/mL
Strenotrophomonas maltophila MIC0.4 mg/mL; MBC 0.8 mg/mL
Bacillus subtilis MICs 5 ppm of AR [9] Candida albicans 475/15 MIC 0.1 mg/mL of RA
MFC 0.2 mg/mL of RA
Bacillus cereus MICs 10 ppm of AR Candida albicans 13/15 MIC 0.1 mg/mL of RA; MFC 0.2 mg/mL of RA
Bacillus polymyxa MICs 15 ppm of AR Candida albicans 17/15 MIC 0.1 mg/mL of RA; MFC 0.2 mg/mL of RA
C. butyricum: C. sporogenes MICs of <20 ppm of RA [18] Candida albicans 527/14 MIC 0.15 mg/mL of RA; MFC 0.3 mg/mL of RA
SARS-CoV-2 IC50 at 25.47 ng μL−1 of RA [25] Candida albicans 10/15 MIC 0.15 mg/mL of RA; MFC 0.3 mg/mL of RA
Enterovirus A71 (EV-A71) In vivo 100 mg/kg/day of RA [26] Candida albicans 532 MIC 0.1 mg/mL of RA; MFC 0.2 mg/mL of RA
S. aureus IZ 22 ± 1.00 mm at 1.33 ± 0.01 mg/g of RA [11] Candida albicans ATCC 10231 MIC 0.2 mg/mL of RA; MFC 0.4 mg/mL of RA
L. monocytogenes IZ 20 ± 2.00 mm at 1.33 ± 0.01 mg/g of RA Candia krusei H1/16 MIC 0.2 mg/mL of RA; MFC 0.4 mg/mL of RA
E. coli IZ 8 ± 0.50 mm at 1.33 ± 0.01 mg/g of RA Candida glabrata 4/6/15 MIC 0.1 mg/mL of RA; MFC 0.2 mg/mL of RA
S. typhimurium IZ 10 ± 0.00 mm at 1.33 ± 0.01 mg/g of RA Candida tropicalis ATCC 750 MIC at 0.2 mg/mL of RA
MFC at 0.4 mg/mL of RA
C. albicans IZ 28 ± 3.00 mm at 1.33 ± 0.01 mg/g of RA Candida parapsilosis ATCC 22019 MIC at 0.1 mg/mL of RA
MFC at 0.2 mg/mL of RA
IZ: Inhibition Zone, MIC:minimal inhibitory concentration. MFC: minimal fungicidal concentration.

3. Antibiofilm Activity

The production of biofilms is one of the main processes responsible for antibiotic resistance. Recent research has revealed that natural substances based on secondary metabolites from plants can prevent the development of biofilms, which are responsible for about 80% of bacterial diseases [27][28]. Biofilms, which are bacterial colonies adhering to the surface and enveloped in a protective extracellular matrix, make bacteria up to 1000 times less susceptible to antibiotics and represent a real health problem [27]. The most common opportunistic fungal diseases in the world are Candida species, which form highly structured biofilms, which are collections of cells of different natures surrounded by an extracellular matrix. Furthermore, the current standard treatment for these infections is to seek innovative treatments for biofilm-related disorders, as these fungal biofilms are typically resistant to conventional antifungal drugs [28].
The quorum sensing inhibition (QSI) potential of rosmarinic acid (RA) towards Aeromonas hydrophila strains MTCC 1739, AH 1, and AH 12 was examined. The A. hydrophila pathogenic strains were isolated from infectious zebrafish species as well as an RA biofilm inhibitory concentration (BIC) versus A. hydrophila strains that was found as 750 μg mL−1. RA at this concentration decreased QS-induced production of hemolysin, elastase, and lipase from A. hydrophila. However, in FT-IR analysis, AR-treated A. hydrophila cells exhibited a reduction in cellular components, and the analysis of gene expression affirmed the negative regulation of virulence genes such as aerA, ahh1, ahyB, and lip. Zebrafish contaminated with A. hydrophila and given RA showed increased survival. Therefore, a study demonstrated the use of RA as an herbal compound to control biofilm formation by QS as well as virulence factor generation in A. hydrophila [29].
Biofilms of C. krusei H1/16 showed the highest resistance against rosmarinic acid treatment; MBEC > 1.6 mg/mL was the minimum biofilm eradication concentration, and biofilms of both C. albicans 475/15 and C. albicans ATCC 10,231 and eradicated with 0.4 mg/mL of rosmarinic acid. In contrast to cell attachment, biofilm formation was more strongly affected for C. albicans strains than for non-C. albicans [22].
RA consumption affected the formation of biofilms at a concentration- and the time-dependent manner, further implying for RA as an effective antimicrobial agent as well as for destroying the activity of planktonic cells and reducing the formation of biofilms at the earlier time stage to their development [30]. RA also inhibits the growth of E. coli K-12 and S. carnosus LTH1502, reducing the density and number of cells [16]. In acidic medium, RA was found to react chemically to nitrite ions to generate 6,6-nitro and 6-dinitrorosmarinic acids, the latter were active at submolecular levels as HIV-1 integrase inhibitors and inhibited viral replication in MT-4 cells, and antiviral effects [20]. RA nitration significantly increased integrase inhibition and antiviral effects without increasing the levels of cellular toxicity. In addition, RA also possesses antimicrobial effects against lactic acid bacteria, yeasts, molds, Enterobacteriaceae spp, and Pseudomonas spp, as well as against psychotropic drugs and L. monocytogenes from chicken meat [31]. In addition, RA exhibits inhibitory effects on the S. aureus cocktail by intimating morphological changes, decreasing and reducing all viable cells, and inducing morphological alterations into cheese and meat samples, from cell shrinkage to the formation of burr-like structures on the cell surface [32][33][34].
The antibacterial effects of rosmarinic acid (RA) against clinical strains of S. aureus from catheter infections were tested by Slobodníková et al. [30]. The regeneration method detected 24 h biofilm eradication activity on microtiter plates. The microtiter plate approach permitted the quantification for biofilm formation activity following application of RA to bacterial samples at 0, 1, 3, and 6 h post biofilm formation, with RA exhibiting antimicrobial activity at concentrations ranging from 625 to 1250 g·mL−1 (MICs equal to MBCs). In the concentration of the 156 to 5000 g·mL−1 evaluated range, there were no biofilm eradicating actions on the 24 h biofilm. When processed at the beginning of biofilm formation, RA subinhibitory doses inhibited the synthesis of biofilm; in concentrations less than the subinhibitory level, the formation of biofilm mass was increased in a time- and concentration-dependent manner. This evidence indicates the potential for RA to be an effective topical antimicrobial agent for treating catheter-related infections, with activity against both planktonic forms of bacteria and inhibitory activity during the early stages of biofilm development. However, it is not practical to use RA as the only agent to treat catheter-related infections [35].

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  15. Suriyarak, S.; Bayrasy, C.; Schmidt, H.; Villeneuve, P.; Weiss, J. Impact of fatty acid chain length of rosmarinate esters on their antimicrobial activity against Staphylococcus carnosus LTH1502 and Escherichia coli K-12 LTH4263. J. Food Prot. 2013, 76, 1539–1548.
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Subjects: Medicine, Research & Experimental
Contributors MDPI registered users' name will be linked to their SciProfiles pages. To register with us, please refer to https://encyclopedia.pub/register :
Ourdia-Nouara Kernou
,
Zahra Azzouz
,
Khodir Madani
,
Patrícia Rijo
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Update Date: 16 Jun 2023
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