Antibacterial Activity of Some Iranian Plant Essential Oils as Disinfectant Agents on Surfaces Contaminated with Staphylococcus Aureus and Pseudomonas Aeruginosa

Antibacterial Activity of Some Iranian Plant Essential Oils as Disinfectant Agents on Surfaces Contaminated with Staphylococcus Aureus and Pseudomonas Aeruginosa | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Antibacterial Activity of Some Iranian Plant Essential Oils as Disinfectant Agents on Surfaces Contaminated with Staphylococcus Aureus and Pseudomonas Aeruginosa Zahra Bastani Nejad, Laila Nikrouz, Abbas Abdollahi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2467112/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The component and antibacterial effects of six essential oils )EOs(, including Cuminum cyminum (CCEO), Artemisia sieberi (ASEO), Laurus nobilis (LNEO), Ferula Gummosa essential (FGEO), Lippia citriodora (LCEO), Cymbopogon citratus (CIEO) were measured by GC-MS and 96-well micro-plates (IC50), against Staphylococcus aureus and Pseudomonas aeruginosa . Then, the antibacterial effects of FGEO, the most efficient EO, were examined on the trolley surface in hospital for 1, 3, 5 and 10 min intervals. While CCEO, ASEO, and FGEO have the highest effects on the growth of S. aureus , CIEO and LNEO showed the best outcomes on P. aeruginosa . In addition, our finding showed that FGEO relatively decreases the growth of S. aureus and P. aeruginosa on the trolley surface (P < 0.05). In addition, FGEO, one of the best disinfectant agents of the present study, showed relatively antibacterial effects on the surface of the trolley. Biological sciences/Biotechnology Biological sciences/Chemical biology Biological sciences/Microbiology Essential oil Antibacterial activity Microdilution Disinfectant Figures Figure 1 Introduction Nosocomial infections (NIs) are acquired infections that are present during the healthcare procedure and do not appear before the hospitalization. In recent years, the prevalence of various NIs mostly grew in America (3.2%) and Europe (5.7%) 1 . The last reports showed that the mean prevalence of NIs in Iranian hospitals (0.71%) is significantly less than worldwide statistics, but some reports demonstrated that the prevalence of NIs in some Iranian hospitals and other Asian clinics would reach up to 9.1%. On the other hand, NIs became one of the main causes of death in the last decades 2 . Multiple studies reported that different microorganisms would cause NIs; but, among them, some bacterial species, such as Staphylococcus aureus ( S. aureus ) and Pseudomonas aeruginosa ( P. aeruginosa ), as significant part of NIs 3 . S. aureus is one of the most common reasons for surgical infections. Moreover, both S. aureus and P. aeruginosa would play the main role in Central Line-Associated Blood Stream Infections 4 . In recent years, we faced new difficulties in controlling NIs. Studies on resistant microorganisms demonstrated that more than 60% of different pathogenic bacteria would present significant resistance to routine antibiotics 5 . In the last 10 years, the increasing growth of resistant S. aureus and other microbial agents in NIs cases has demanded new valuable agents to protect hospitalized patients 6 . On the other hand, the resistant serotype of P. aeruginosa had increased recently and showed remarkable resistance against gentamicin and ciprofloxacin. Besides, common antimicrobial agents have had low efficiency in controlling the growth of NIs in recent years. The last evaluations revealed that the resistant gram-positive and gram-negative bacterial agents developed in the Iranian hospitals and we require novel agents to prevent NIs morbidity and mortality 7 . In the last years, numerous herbal products with commercial uses as well as sedative and antioxidant properties have become available worldwide. Therefore, we can use these products without worrying about consumer rejection. On the other hand, herbal extracts show more antimicrobial potentials with the least adverse effects 8 . Studies showed that herbal essential oils (EOs) and extracts with antimicrobial properties would prevent the activities of different bacterial agents 9 . Recently, novel studies revealed several herbal extracts with promising antimicrobial effects on different microorganisms and provided new complementary treatments with the least side effects 10 . Moreover, several recent studies provided multiple antimicrobial products from Iranian herbal species 11 . Various studies have shown that the use of plant EOs has encouraging antibacterial effects. Today, a large number of plant essential oils with significant antimicrobial effects, including Myrtle communis , Thymus vulgaris , Lavandula angustifolia , and Mentha pulegium , are known to have good uses in suppressing bacterial growth 12 . On the other hand, although several chemical disinfectant agents have been developed and achieve acceptable level of disinfection, some practices showed that these agents would show multiple bacterial resistance 13 . Using plant EOs in cleansing gels showed similar effectiveness to previous synthetic products with fewer side effects. Therefore, the present study aimed to evaluate the antibacterial activity of different herbal extracts on S. aureus and P. aeruginosa . Then, the most effective EOs were applied on the hospital surfaces contaminated with S. aureus and P. aeruginosa to achieve novel natural agents to restrict NIs in the future. Results Chemical composition Table 1. Shows the results of GC-MS analysis of FGEO (the plant with the highest effect). The analysis showed that FGEO mainly contains β-Pinene. It seems that this combination is the most effective one based on the high amount of β-pinene compound and based on the results of other articles. Table 1. The components of Ferula Gummosa essential oil No RT % Components KI Type 1 11.36 4.08 α-pinene 935 MH 2 13.44 0.44 sabinene 976 MH 3 13.77 58.70 ẞ-pinene 983 MH 4 14.27 2.14 myrcene 993 MH 5 15.26 1.26 3-carene 1012 MH 6 16.40 0.64 limonene 1034 MH 7 16.51 0.40 ẞ-phellandrene 1036 MH 8 16.74 0.67 trans- ẞ-ocimene 1040 MH 9 24.34 0.57 trans-galbanolene 1192 Other 10 37.78 0.82 alloaromadendrene 1492 SH 11 40.52 0.88 elemol 1561 SO 12 42.43 6.52 guaiol 1609 SO 13 44.80 1.10 ẞ-eudesmol 1673 SO 14 45.09 6.78 bulnesol 1680 SO 84.98 Total identified Abbreviations. RT, Retention time; KI, Kovalts index; MH, Monoterpene Hydrocarbons; SH, Sesquiterpene Hydrocarbons; SO, Oxygenated Sesquiterpenes Antibacterial effects Fig. 1a shows the antibacterial effects of various concentrations (0.03-8.00 mg/mL) of Cuminum cyminum essential oil (CCEO). The highest effects were reported against standard S. aureus (SSA, 32%) and clinical P. aeruginosa (CPA, 47%) at 8.00 mg/mL CCEO. In addition, this EO decreased the growth of clinical S. aureus (CSA) and standard P. aeruginosa (SPA) with fewer significant antibacterial effects (55% and 56%). The effect of Artemisia sieberi essential oil (ASEO) at different concentrations on the targeted bacterial growth is reported in Fig. 1b. ASEO mostly restricted the SSA growth (20%), while other results showed lower antibacterial effects (CSA: 42%; SPA: 38%; CPA: 43%). As shown in Fig. 1c, Laurus nobilis essential oils (LNEO) significantly inhibited the growth of SSA (39%), SPA (35%), and CPA (38%). Moreover, LNEO decreased the activity of CSA with fewer effects (51%). The antibacterial effects of FGEO were observed in Fig. 1d. Totally, FGEO showed one of the best results in the present study and remarkably inhibited all spp. While the growth of CSA was decreased to 17%, the growth of SSA, SPA, and CPA was reduced to 39%, 36%, and 41%. Lippia citriodora essential oils (LCEO) showed significant inhibitory effects with increasing the concentration (Fig. 1e). The growth of all targeted bacteria was inhibited. LCEO at a concentration of 8.00 mg/mL reduced the growth of CPA and CSA to 44% and 39%. On the other hand, the growth restriction of standard spp. higher than in clinical specimens (SPA, 33%; SSA, 37%). Results of bacterial inhibitory effects of Cymbopogon citratus essential oil (CIEO) are shown in Fig. 1f. Mostly, CIEO showed antibacterial activity on SPA (27%), but CIEO totally showed significant inhibitory effects (more than 50% bacterial growth inhibition) on all the other spp. (SSA, 30%; CSA, 32%; CPA, 38%). Disinfection activity of FGEO on trolley surface Table 2 shows the antibacterial effects of FGEO on the surface of trolley contaminated with standard and clinical S. aureus and P. aeruginosa . FGEO significantly decreased the growth of bacterial agents in all samples. In addition, the antibacterial effects were positively related to the concentration and the time of exposure. Table 2 . antibacterial effects of FGEO on trolley surface contominated with S. S. aureus, C.S.aureus, S.P.aeroginosa, C.P.aeroginosa. Groups Positive control 4000 mg/mL 8000 mg/mL Negative control Time of exposure to FGEO (minutes) 1 3 5 10 1 3 5 10 1 3 5 10 1 3 5 10 The mean number of S.S. aureus colonies 10 6 10 6 10 6 10 6 10 4 10 3 10 3 10 3 10 3 10 2 10 2 10 2 0 0 0 0 The mean number of C.S. aureus colonies 10 6 10 6 10 6 10 6 10 4 10 3 10 3 10 3 10 3 10 2 10 2 10 2 0 0 0 0 The mean number of S.P. aeruginosa colonies 10 6 10 6 10 6 10 6 10 5 10 4 10 3 10 3 10 4 10 4 10 3 10 2 0 0 0 0 The mean number of C.P. aeruginosa colonies 10 6 10 6 10 6 10 6 10 5 10 4 10 3 10 3 10 4 10 4 10 3 10 2 0 0 0 0 Abbreviations: positive control, washed with normal saline; negative control, washed with alcohol; FGEO, Ferula Gummosa essential oil; S.S. aureus, Standard Staphylococcus aureus ; C.S. aureus, clinical Staphylococcus aureus ; S.P. aeruginosa, Standard pseudomonas aeruginosa ; C.P. aeroginosa, clinical pseudomonas aeruginosa . Discussion The present study aimed to evaluate the antibacterial effects of six EOs at the various concentrations on standard and clinical species of two common nosocomial pathogens ( S. aureus and P. aeruginosa ). In addition, we selected the most efficient antibacterial EO to investigate its antibacterial effects on nosocomial surfaces. Generally, S. aureus is found approximately all over the environment, especially on the skin and nasal cavity. On the other hand, S. aureus is one the most opportunistic pathogens that would result in several nosocomial infections 14 . The results showed that ASEO had the best antibacterial effects on SSA among examined EOs. In the following, CCEO and CIEO represented acceptable effects, but the other EOs had the lower inhibitory activity on SSA. P. aeruginosa is another opportunistic pathogen that belongs to gram-negative strains. The genus Pseudomonas is available to survive in various difficult environments and causes life-threatening nosocomial infections. Totally, the experimented EO in the present study showed fewer antibacterial effects against P. aeruginosa . The best findings were observed in the case of CIEO, but the other EOs showed significant inhibitory effects, except CCEO 15 . The genus Artemisia is a well-known medicinal plant that grows worldwide 16 . Artemisia sieberi is a special species of this large genus usually found in the northern regions of Iran and Saudi Arabia. Chemical analysis revealed that ASEO is mostly made up of Artemisia ketone and 1,8-cineole. Moreover, Vahdani et al. reported that the main compounds of ASEO were piperitone, camphor, and 1, 8-cineole 17 . On the other hand, a recent GC-MS indicated that Camphor, 1,8-cineole, and β-Thujone were the first major components of ASEO 18 . Therefore, we can conclude that 1,8-cineole is an improved component in the chemical composition of ASEO and would be one of the major antibacterial agents of this EO. Different species of genus Artemisia have encouraging antibacterial activities 19 . Also, A. sieberi products indicated great inhibitory effects on both Gram-negative and Gram-positive strains 20 . In 2013, Hedayati rad et al. conducted a novel Pullulan film incorporated with ASEO to evaluate its protective effects against different bacterial pathogens. This study revealed that ASEO at low concentrations (5%) has no inhibitory effects on bacterial growth, but with increasing the concentration (at least to 10%) the antibacterial effects would develop. The highest inhibitory zone (18.52 ± 0.73) was observed at 35% 21 . Pure ASEO made large inhibitory zones against S. aureus and P. aeruginosa in Muller Hinton agar plates. Besides, incorporating basal seed gum films with ASEO resulted in a big inhibitory zone (23.2 ± 0.7 mm) on plates infected with S. aureus and a smaller inhibitory zone (10.2 ± 0.6 mm) in the case of P. aeruginosa 22 . Supporting the results of the present study, others also reported that ASEO represented valuable MIC (10 µL) and minimal bactericidal concentration (MBC) (20 µL) against S. aureus 17 . Moreover, recent studies on synergic antibacterial effects of ASEO combined with different routine antibiotics demonstrated that ASEO would increase the inhibitory effects of ampicillin, oxacillin, cephalothin, piperacillin, and vancomycin on S. aureus . On the other hand, the combination of various antibiotics, such as piperacillin, gentamicin, ampicillin, and erythromycin with ASEO indicated significant synergic effects on the growth of P. aeruginosa 23 . Therefore, using pure ASEO and its combination with different products would provide novel antibacterial materials that represent higher inhibitory effects and decrease the difficulties of previous routine antibiotics. Chemical composition analysis indicated that CCEO mostly contains 1-phenylpropanol and gamma-Terpinene 24 . Besides, Hajlaoui et al. reported that CCEO mainly comprised cumin aldehyde and gamma-terpinene. On the other hand, the first evaluations revealed that Isobutyl isobutyrate, a-Thujene, and a-Pinene are the most common compositions in CCEO. CCEO is believed to have a protective role in food production and storage industries 25 . Further studies demonstrated that the most protective CCEO concentrations will appear at 0.03% that prevented the S. aureus growth in feta cheese and infected hamburgers 24 . CCEO demonstrated promising MBC and MIC on SSA (2.5 to 5 µg/mL, and 1.25 to 5 µg/mL). Moreover, this study showed that CCEO had the same antibacterial effect on SSA as vancomycin, a highly used antibiotic with several adverse effects 26 . On the other hand, some other studies supported our results. Saee et al. reported that the extract and EO from C. cyminum significantly diminished the CPA with higher efficiency than several antibiotics such as gentamicin, amoxicillin, and clotrimazole 27 . Therefore, utilizing CCEO in preventing and treating procedures would decrease the limitations of routine antibiotics in the future, but the controversies of its antibacterial effects on gram-negative strains remained for further studies. C. citratus has a valuable protecting usage in food industries to restrict the activity of common food-borne pathogens. Utilizing CIEO in the preservation of cooked beef patties would save its quality and prevent the growth of pathogenic bacterial strains 28 . The evaluations showed that neral, geranial, limonene, and geranyl acetate are the major components of CIEO. C. citratus products mainly prevent the growth of fungal ( Candida strains) and Gram-positive bacterial species, such as S. aureus , biofilms. Based on the last in vitro trials, the CIEO provides the same antibacterial effects on S. aureus as vancomycin and would be a favorable product to prevent the growth of nosocomial pathogens with fewer adverse effects in the future 29 . In addition, a recent study on EO from the leaves of C. citratus indicated a bigger zone of inhibition in the case of S. aureus (32.0 ± 0.75 mm) and no antibacterial effects on P. aeruginosa . Nevertheless, the other studies showed that C. citratus extracts would represent more antibacterial effects on gram-negative strains such as P. aeruginosa . Subramanian et al. reported that the leaf and root extracts resulted in a very large zone of inhibitions (> 30 mm), and would be encouraging herbal products against these pathogenic gram-negative strains 30 . Moreover, the results of our study represented that CIEO mainly decreased the growth of SPA more than the other targeted strains. Therefore, further studies are required to evaluate these controversial results. L. citriodora and hundreds of other species in the Lippia genus are well-known ancient plants utilized for several decades, especially in Western regions of the world. GC-MS analysis reported that neral and geranial are the major compounds of experimented LCEO 31 . In addition, other studies added terpenic alcohols, monoterpenic hydrocarbons (dl-limonene), and α-curcumene to previously reported chemical components. Several studies reported various medicinal properties, such as sedative, diuretic, and antispasmodic use in recent years. Moreover, the recent papers introduced LCEO as a moderate antibacterial product 32 . Although pure LCEO provided an acceptable zone of inhibition on sterile Whatman paper discs infected with gram-positive ( S. aureus ) and gram-negative ( E. coli ) strains, this EO mainly affects the gram-negative strains more than gram-positive species. On the other hand, some other studies represented controversial results. Evaluating the effects of LCEO on different pathogens showed that this EO has no significant inhibitory or bactericidal effects on P. aeruginosa and the MIC and MBC are higher in the case of E. coli than S. aureus 33 . Therefore, the present study and all other studies introduced LCEO as an encouraging antibacterial product, especially for gram-positive strain, but further studies are required to put an end to the controversial results in the case of P. aeruginosa . Laurus nobilis is another aromatic plant with antimicrobial and flavor properties in food industries. According to the high requirement of enough water and humidity, L. nobilis usually grows in warmer climate places such as the south of the Mediterranean region. Methyl eugenol, 1,8-cineole, and α-terpinyl acetate are the major compounds of LNEO. LNEO has significant inhibitory effects on all gram-positive and -negative strains, like P. aeruginosa , S. aureus , E. coli . The measurements of LNEO inhibition zone showed higher antibacterial effects than tetracycline (8 mg/mL), especially in the case of gram-positive strains. The evaluations revealed that LNEO has high antibacterial effects on gram-positive strains such as S. aureus (inhibition zone = 13 mm), S. intermedius , and K. pneumoniae . Also, LNEO significantly reduces the biofilm activity with acceptable MIC, ranged from 0.3 to 96.4 mg/mL 34 . Further comparisons showed that gram-positive strains are more susceptible to LNEO than P. aeruginosa and E. coli . The recent evaluations demonstrated that minimal inhibitory activities of LNEO would be represented at 0.25 mg/mL. On the other hand, LNEO showed remarkable protective effects against common foodborne pathogens, such as E. coli 35 . The aromatic herbal families, such as Apiaceae , are common plants that grow in different regions of Iran. The genus of Ferula is a medicinal herb mostly found in EMERO and north Africa floras 36 . Several studies revealed different protective effects of F. gummosa against infection, inflammatory and spasmodic processes. EO from F. gummosa seeds mostly showed strong antibacterial effects on gram-positive strains and slight inhibitory activity against gram-negative strains, such as P. aeruginosa . Abbaszadegan et al. reported that β-pinene made up more than 50% of the chemical composition of FGEO. Moreover, FGEO at 50 µg/mL resulted in a big inhibitory zone (22.5 ± 0.82 mm) on plates infected with S. aureus 37 . Considering the antibacterial effects on both S. aureus and P. aeruginosa, our findings showed that FGEO is the most effective agent among the evaluated EOs in the present study. As a result, we selected FGEO to investigate its disinfection activity on the surface of emergency hospital. In the last two decades, several studies have attempted to use EO-based disinfectants instead of conventional disinfectants. Sharma et al. indicated that a novel green disinfectant, named Neutral Biodegradable Disinfectant (BND) would appropriately clear the contaminated surfaces. Their findings confirmed that phenol-based disinfectants and NBD products disinfect environmental microorganisms equally 38 . As a result, given that natural products provide much less resistance and reduce the population of microorganisms evenly, many of these products are expected to replace conventional phenol-based disinfectants in the coming years 39 . Increasing bacterial resistance to conventional disinfectants resulted in a great challenge in different healthcare services, especially in dental procedures 40 . In recent years, several observations indicated that green disinfectants would become one of the main agents to provide streel dental practices. In 2019, Nausheen et al. confirmed that aloe vera gel at high concentrations (90%) is a safe disinfectant and shows less poor outcomes than conventional chemical products 41 . In addition, our findings supported the previous results. FGEO relatively could decrease the growth of both bacterial agents on the trolley surface. Although the increasing concentration of FGEO developed its antibacterial activity, a significant population of bacterial colonies grew on the evaluated samples of our study. Therefore, the application of FGEO-based disinfectants would not be able to completely clear nosocomial surfaces, similar to conventional and novel green disinfectants 42 . The present study has several strengths and limitations. The main strength of our study was the comparison of the antibacterial effects of above-mentioned EOs on clinical and standard samples of two major gram-positive and gram-negative strains. Based on our expanded literature review on previous studies, we mainly hypothesized that the gram-negative strains and clinical samples would represent more resistance against different antibacterial EOs. Generally, the findings supported these hypotheses with little controversy (especially in the case of CIEO and LNEO). Moreover, we faced some conflicts in comparison with previous studies. On the other hand, we just reported the inhibition activity of EOs on bacterial growth in percent. Therefore, further studies with more targeted pathogens and expand measurements would be required to collect more sufficient data and resolve observed controversies. Methods Bacterial species In this study, we obtained standard bacterial species from the Microbiology laboratory of Pasteur Institute of Tehran (Standard species of S. aureus , ATCC 25923; Standard species of P. aeruginosa , ATCC 27853). On the other hand, we used clinical isolated Methicillin-resistant S. aureus (MRSA) and P. aeruginosa Metallo-beta-lactamase (MBL)-producing from wound infection from previous studies of our researches. Also, we utilized molecular method (PCR test) and the routine culture media (the disk-diffusion method and minimal inhibitory concentration (MIC) test) tests to identify phenotypically and genotypically MBL-producing P. aeruginosa and MRSA 43 . We obtained all EOs from Tabib Daroo Company (a pharmaceutical company in Kashan). Herbarium specifications of each plant are shown in the Table 3. Table 3. Herbarium specifications of applied plants No Scientific names (Family) Part used Herbarium code Collector Place of collection 1 Cuminum cyminum (Cuminum Family) Seed IAUM- 211 Dr. A. Ziaeei Bajestan, khorasan, Iran 2 Artemisia sieberi (Asteraceae Family) Leave HUMZ-8629 Dr. A. Taheri Ghaemshahr, Mazandaran, Iran 3 Laurus nobilis (Lauraceae Family) Leave IAUGH-1409 Dr. E. Bahrani Tehran, Tehran, Iran 4 Ferula gummosa (Apiaceae Family) All UQH-1530 Dr. A. Khani Arak, Markazi, Iran 5 Lippia citriodora (Verbenaceae Family) Leave KF-1513 Dr. Mehrabani Sirch, Kerman, Iran 6 Cymbopogon citratus (Poaceae Family) Leave HUI-TB400CI A. Mazaheri Kashan, Isfahan, Iran Bacterial culture We used Muller Hinton broth, containing 0.5% DMSO for bacterial growth (Merck Chemicals, Germany) 44 . Gas Chromatography/Mass Spectrometry analysis GC-MS analysis was deployed to identify the compounds of the selected herbal EOs. We utilized a 7890A Network GC system alongside a 5975A massive detector (Agilent Technologies, Santa Clara, CA, USA). The compounds of EOs were separated via fused columns of HP-5MS silica (length, 30 meters; internal diameters, 250 µM; film thickness, 25 µM). The temperature was set first at 50°C, increased gradually to 200°C (5°C per minute), and remained at this temperature for 1 minute. Then, we elevated the temperature up to 250°C (10°C per minute) and again increased to 300°C with higher rate (25°C per minute). On the other hand, helium was utilized to carry the ingredients. At last, we analyzed the components based on our previously described method 45 . Determination of the antibacterial activity of Eos We utilized a 96-well plate to provide a proper situation for evaluation of antibacterial effects of EOs against purposed bacterial species based on standard protocols 46 . We achieved 1.5 × 10 8 CFU/mL of bacterial colonies to pass the 0.5 McFarland turbidity. Then 20-µL suspension was added to all wells. After preparation of bacterial agents, we dissolved EOs in Muller Hinton Broth to provide a standard dilution series (8.00, 4.00, 2.00, 1.00, 0.50, 0.25, and 0.13 mg. mL-1). After described dilution, we added 80 µL of broth suspension to all wells and incubated in standard set (24h incubation at 37°C). The turbidity evaluation was performed at 630 nm via plate reader (Synergy HTX Multi-Mode Reader, USA) and the bacterial growth was assessed through Equation 1. Measurement of disinfection properties of Ferula Gummosa essential oil (FGEO) FGEO were prepared in two different concentrations (4000 and 8000 µg) to measure its disinfection activity on 72 samples (3 × 3 cm 2 ) from different emergency trolley steel surfaces (three times). First, we cleaned the samples using 70% ethyl alcohol and sterile cotton soaked in sterile water (3 times, each time from the center to the outside). Then, all samples were contaminated with bacterial agents by streel swab and they were allowed to drain at room temperature. The bacterial strains were cultured in Muller Hinton Agar (MHA) and suspension of 0.5 McFarland for 24 to 48 hours (approximately 1.5 × 10 8 bacterial colonies). Finally, all surfaces were divided into four groups (alcohol as negative control, untreated surface as positive control, 4000 FGEO, and 8000 FGEO). Each group divided into four subgroups of bacterial species ( S. aureus and P. aeruginosa ). The final samples were gathered after 1, 3, and 5 minutes. The samples were carried to the laboratory in a freezer to prevent probable growth. Each sample vortexed for 1 minute and 100 µL of the sample was cultured on blood agar. The broths were kept in an incubator for 24 hours at 37℃. Finally, the bacterial colonies of each sample were counted and the mean was reported 13 . Statistical methods We carried out antibacterial tests in triplicates. SPSS software (Version 22, SPSS Inc, USA) was utilized to measure the means ± standard deviations and draw charts. We deployed a regression model (performed in CalcuSyn software, Free version, BIOSOFT, UK) to calculate IC50 of all Eos; then, we compared all IC50 of the Eos together via one-way ANOVA using SPSS software (Version 22, SPSS Inc, USA) independent sample t-test. The confidence interval of this study was set at 95% (CI 95%). Conclusion This study indicated that all experimented EOs has encouraging antibacterial effects against targeted pathogens. Some products, such as CCEO, ASEO, and FGEO represented higher inhibitory effects on S. aureus and the latter, including CIEO and LNEO, had better effects on P. aeruginosa . Therefore, further studies are required to evaluate the antibacterial effects of the combination of these EOs and utilize novel techniques, such as nanoparticles to develop more efficient natural products and prevent life-threatening nosocomial infections. Since essential oils can have synergistic or antagonistic effects on each other, it is suggested to investigate the effects of the combination of essential oils. Nonetheless, it will be considered in our additional studies in the future. Declarations Acknowledgements The authors wish to acknowledge the support of Fasa University of Medical Sciences for facilitating this research. Author contributions All authors contributed to the study conception and design. Funding This work was supported by a grant from the Fasa University of Medical Sciences. Data Availability The authors confirm that the data supporting the findings of this study are available within the article. Conflict of interest statement All other authors have no conflicts of interest to declare. References Suetens, C. et al. 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The evaluation of chemical composition, antimicrobial activity and drug interaction in the essential oil of Artemisia sieberi. Rezai, R., Sadeghi, E., Nateghi, L. & Mohammadi, M. The effect of Cuminum cyminum essential oil on growth and survival of Staphylococcus aureus during storage of hamburger. International Journal of Biosciences 5 , 18–26 (2014). Wongkattiya, N., Sanguansermsri, P., Fraser, I. H. & Sanguansermsri, D. Antibacterial activity of cuminaldehyde on food-borne pathogens, the bioactive component of essential oil from Cuminum cyminum L. collected in Thailand. Journal of Complementary and Integrative Medicine 16 (2019). Sharifi, A., Mohammadzadeh, A., Salehi, T. Z., Mahmoodi, P. & Nourian, A. Cuminum cyminum L. Essential Oil: A Promising Antibacterial and Antivirulence Agent Against Multidrug-Resistant Staphylococcus aureus. Frontiers in Microbiology 12 (2021). Saee, Y. et al. Evaluation of antimicrobial activity of Cuminum cyminum essential oil and extract against bacterial strains isolated from patients with symptomatic urinary tract infection. Novelty in Biomedicine 4 , 147–152 (2016). Siewe, F. B., Mbougueng, P. D., Tatsadjieu, L. N., Noumo, T. N. & Mbofung, C. M. The potential application of syzygium aromaticum and Cymbopogon citratus essential oils as natural preservatives of beef patties. Food and Nutrition Sciences 6 , 374 (2015). Oliveira, J. B. et al. In vitro and in vivo antimicrobial activity of Cymbopogon citratus (DC.) Stapf. against Staphylococcus spp. isolated from newborn babies in an intensive care unit. Microbial Drug Resistance 25 , 1490–1496 (2019). Subramaniam, G., Yew, X. Y. & Sivasamugham, L. A. Antibacterial activity of Cymbopogon citratus against clinically important bacteria. South African Journal of Chemical Engineering 34 , 26–30 (2020). Fitsiou, E. et al. Chemical composition and evaluation of the biological properties of the essential oil of the dietary phytochemical Lippia citriodora. Molecules 23 , 123 (2018). Benoua, F. Z. et al. Antimicrobial activity of the thio-cyclized Lippia citriodora leaf essential oil cultivated in Algeria. Journal of Biologically Active Products from Nature 9 , 250–259 (2019). Bensabah, F., Sbayou, H., Amghar, S., Lamiri, A. & Naja, J. Chemical composition and antibacterial activity of essential oils of two aromatic plants: Mentha spicata and Lippia citriodora irrigated by urban wastewater. Int J Eng 2 , 1560–1569 (2013). Merghni, A., Marzouki, H., Hentati, H., Aouni, M. & Mastouri, M. Antibacterial and antibiofilm activities of Laurus nobilis L. essential oil against Staphylococcus aureus strains associated with oral infections. Current Research in Translational Medicine 64 , 29–34 (2016). da Silveira, S. M. et al. Chemical composition and antibacterial activity of Laurus nobilis essential oil towards foodborne pathogens and its application in fresh Tuscan sausage stored at 7 C. LWT-Food Science and Technology 59 , 86–93 (2014). Sahebkar, A. & Iranshahi, M. Biological activities of essential oils from the genus Ferula (Apiaceae). Asian Biomed 4 , 835–847 (2010). Abbaszadegan, A. et al. Antimicrobial and cytotoxic activity of Ferula gummosa plant essential oil compared to NaOCl and CHX: a preliminary in vitro study. Restorative dentistry & endodontics 40 , 50–57 (2015). Sharma, V. & Agarwal, S. Comparative efficacy of an Essential Oil based (Green) disinfectant with a Conventional Phenol based disinfectant. Dhakad, A. K., Pandey, V. V., Beg, S., Rawat, J. M. & Singh, A. Biological, medicinal and toxicological significance of Eucalyptus leaf essential oil: a review. Journal of the Science of Food and Agriculture 98 , 833–848 (2018). Bagheri, S. M. et al. Evaluation of disinfection quality of Dental Faculty Units of Ahvaz Jundishapur University of Medical Sciences, Southwest of Iran in 2017. J Mol Biol Res 8 , 88–94 (2018). Nausheen, A. et al. Effect of Different Chemical and Herbal Disinfectant Solutions on the Mechanical and Physical Properties of Gutta-percha: An In Vitro Study. Journal of Operative Dentistry and Endodontics 4 , 84–87 (2019). Amini, K., Soltani Gerdfaramarzi, M., Mokhtari, A. & Amini, P. Evaluate of the Effects of three Antiseptic Solutions (Septisurface, DDSH and H2O2) on Contamination Levels of Restorative Dental Unit. Journal of Mashhad Dental School 39 , 303–314 (2015). Qasemi, H. et al. Promising antibacterial effect of impregnated nanofiber mats with a green nanogel against clinical and standard strains of Pseudomonas aeruginosa and Staphylococcus aureus. Journal of Drug Delivery Science and Technology 66 , 102844 (2021). Abdollahi, A., Zarenezhad, E., Osanloo, M., Ghaznavi, G. & Khalili Pour, M. Promising antibacterial activity of a mat of polycaprolactone nanofibers impregnated with a green nanogel. Nanomedicine Research Journal 5 , 192–201 (2020). Osanloo, M. et al. Chitosan nanocapsules of tarragon essential oil with low cytotoxicity and long-lasting activity as a green nano-larvicide. J Nanostruct 9 , 723–735 (2019). Valizadeh, A., Shirzad, M., Esmaeili, F. & Amani, A. Increased antibacterial activity of cinnamon oil microemulsionin comparison with cinnamon oil bulk and nanoemulsion. Nanomedicine Research Journal 3 , 37–43 (2018). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2467112","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":171263165,"identity":"7dacc0a5-5072-44a6-93e2-c5427bdd1fd6","order_by":0,"name":"Zahra Bastani Nejad","email":"","orcid":"","institution":"Fasa University of Medical Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zahra","middleName":"Bastani","lastName":"Nejad","suffix":""},{"id":171263167,"identity":"79f0e6f5-8d30-4628-a14f-22f98f9df764","order_by":1,"name":"Laila Nikrouz","email":"","orcid":"","institution":"Fasa University of Medical Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Laila","middleName":"","lastName":"Nikrouz","suffix":""},{"id":171263168,"identity":"b83d2e13-a90f-43f0-a6dc-8f6fc8831d16","order_by":2,"name":"Abbas Abdollahi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYDACCQYDhgcMB3gYGIDoA1CAjZ0YLQlQLYwzQFqYidTCANLCzAMSIaSFf3bzxgcJNXdkGNjPHv5s82ubPB8zA+OHjzl4LLlzrNgg4dgzHgaevDTp3L7bhm3MDMySM7fhseZGjplEAtthoJNyzJhze24zArWwMfPi0SIP1vIPqIX/jfFny57b9gS1GIC0JLYBtUjkGEgz/LidSFCLIcgviX2Hedgk3qVJ9jbcTm5jZmzG6xe528AQ+/DtsD0/f+7hDz/+3Lad39588MNHfN6HATYQwdgGJhuIUA8Hf0hRPApGwSgYBSMFAABu0U/5H5RyXAAAAABJRU5ErkJggg==","orcid":"","institution":"Fasa University of Medical Sciences","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Abbas","middleName":"","lastName":"Abdollahi","suffix":""}],"badges":[],"createdAt":"2023-01-11 12:14:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2467112/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2467112/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":32276878,"identity":"37c99c06-aefd-4049-8de2-12ac037a597e","added_by":"auto","created_at":"2023-01-31 15:30:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":95201,"visible":true,"origin":"","legend":"\u003cp\u003eThe antibacterial effects off essential oil. S.S. aureus, Standard \u003cem\u003eStaphylococcus aureus\u003c/em\u003e; C.S. aureus, clinical \u003cem\u003eStaphylococcus aureus\u003c/em\u003e; S.P. aeruginosa, Standard \u003cem\u003epseudomonas aeruginosa\u003c/em\u003e; C.P. aeroginosa, clinical \u003cem\u003epseudomonas aeruginosa\u003c/em\u003e. A) \u003cem\u003eCuminum cyminum\u003c/em\u003e essential oil. b)\u003cem\u003e Artemisia sieberi\u003c/em\u003e essential. c) \u003cem\u003eLauruse Nobilis \u003c/em\u003eessential oil. d) \u003cem\u003eFerula Gummosa\u003c/em\u003e essential oil. e) \u003cem\u003eLippia Citriodora\u003c/em\u003e essential oil. f)\u003cem\u003e Cymbopogon Citratus\u003c/em\u003e essential oil\u003c/p\u003e","description":"","filename":"F1.png","url":"https://assets-eu.researchsquare.com/files/rs-2467112/v1/4d2816b830811b24b9957dd0.png"},{"id":33441942,"identity":"eda7a9cb-a6b0-409c-b628-09251d27036c","added_by":"auto","created_at":"2023-02-25 10:00:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":482404,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2467112/v1/185ff7d1-0a5e-4dd4-a0d1-717f0de4b6c8.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antibacterial Activity of Some Iranian Plant Essential Oils as Disinfectant Agents on Surfaces Contaminated with Staphylococcus Aureus and Pseudomonas Aeruginosa","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNosocomial infections (NIs) are acquired infections that are present during the healthcare procedure and do not appear before the hospitalization. In recent years, the prevalence of various NIs mostly grew in America (3.2%) and Europe (5.7%) \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. The last reports showed that the mean prevalence of NIs in Iranian hospitals (0.71%) is significantly less than worldwide statistics, but some reports demonstrated that the prevalence of NIs in some Iranian hospitals and other Asian clinics would reach up to 9.1%. On the other hand, NIs became one of the main causes of death in the last decades \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Multiple studies reported that different microorganisms would cause NIs; but, among them, some bacterial species, such as \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (\u003cem\u003eS. aureus\u003c/em\u003e) and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e (\u003cem\u003eP. aeruginosa\u003c/em\u003e), as significant part of NIs \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. \u003cem\u003eS. aureus\u003c/em\u003e is one of the most common reasons for surgical infections. Moreover, both \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e would play the main role in Central Line-Associated Blood Stream Infections \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn recent years, we faced new difficulties in controlling NIs. Studies on resistant microorganisms demonstrated that more than 60% of different pathogenic bacteria would present significant resistance to routine antibiotics \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. In the last 10 years, the increasing growth of resistant \u003cem\u003eS. aureus\u003c/em\u003e and other microbial agents in NIs cases has demanded new valuable agents to protect hospitalized patients \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. On the other hand, the resistant serotype of \u003cem\u003eP. aeruginosa\u003c/em\u003e had increased recently and showed remarkable resistance against gentamicin and ciprofloxacin. Besides, common antimicrobial agents have had low efficiency in controlling the growth of NIs in recent years. The last evaluations revealed that the resistant gram-positive and gram-negative bacterial agents developed in the Iranian hospitals and we require novel agents to prevent NIs morbidity and mortality \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn the last years, numerous herbal products with commercial uses as well as sedative and antioxidant properties have become available worldwide. Therefore, we can use these products without worrying about consumer rejection. On the other hand, herbal extracts show more antimicrobial potentials with the least adverse effects \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Studies showed that herbal essential oils (EOs) and extracts with antimicrobial properties would prevent the activities of different bacterial agents \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eRecently, novel studies revealed several herbal extracts with promising antimicrobial effects on different microorganisms and provided new complementary treatments with the least side effects \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Moreover, several recent studies provided multiple antimicrobial products from Iranian herbal species \u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Various studies have shown that the use of plant EOs has encouraging antibacterial effects. Today, a large number of plant essential oils with significant antimicrobial effects, including \u003cem\u003eMyrtle communis\u003c/em\u003e, \u003cem\u003eThymus vulgaris\u003c/em\u003e, \u003cem\u003eLavandula angustifolia\u003c/em\u003e, and \u003cem\u003eMentha pulegium\u003c/em\u003e, are known to have good uses in suppressing bacterial growth \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. On the other hand, although several chemical disinfectant agents have been developed and achieve acceptable level of disinfection, some practices showed that these agents would show multiple bacterial resistance \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Using plant EOs in cleansing gels showed similar effectiveness to previous synthetic products with fewer side effects. Therefore, the present study aimed to evaluate the antibacterial activity of different herbal extracts on S. aureus and \u003cem\u003eP. aeruginosa\u003c/em\u003e. Then, the most effective EOs were applied on the hospital surfaces contaminated with \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e to achieve novel natural agents to restrict NIs in the future.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eChemical composition\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 1. Shows the results of GC-MS analysis of FGEO\u0026nbsp;(the plant with the highest effect). The analysis showed that FGEO mainly contains \u0026beta;-Pinene.\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003eIt seems that this combination is the most effective one based on the high amount of \u0026beta;-pinene compound and based on the results of other articles.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e The components of Ferula Gummosa essential oil\u003c/p\u003e\n\u003ctable border=\"0\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003e\u003cstrong\u003eComponents\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e\u003cstrong\u003eKI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e\u003cstrong\u003eType\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e11.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e4.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003e\u0026alpha;-pinene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e935\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e13.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003esabinene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e976\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e13.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e58.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eẞ-pinene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e983\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e14.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e2.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003emyrcene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e993\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e15.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e1.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003e3-carene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e16.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003elimonene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1034\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e16.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eẞ-phellandrene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1036\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e16.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003etrans- ẞ-ocimene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1040\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eMH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e24.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003etrans-galbanolene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1192\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e37.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003ealloaromadendrene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1492\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eSH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e40.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e0.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eelemol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1561\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eSO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e42.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e6.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eguaiol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1609\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eSO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e44.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e1.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eẞ-eudesmol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1673\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eSO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e45.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e6.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003ebulnesol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e1680\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003eSO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e84.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.857142857142854%\"\u003e\n \u003cp\u003eTotal identified\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.224489795918368%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.26530612244898%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations. RT, Retention time; KI, Kovalts index; MH, Monoterpene Hydrocarbons; SH, Sesquiterpene Hydrocarbons; SO, Oxygenated Sesquiterpenes\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial effects\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFig. 1a shows the antibacterial effects of various concentrations (0.03-8.00 mg/mL) of \u003cem\u003eCuminum cyminum\u003c/em\u003e essential oil (CCEO). The highest effects were reported against standard \u003cem\u003eS. aureus\u003c/em\u003e (SSA, 32%) and clinical \u003cem\u003eP. aeruginosa\u003c/em\u003e (CPA, 47%) at 8.00 mg/mL CCEO. In addition, this EO decreased the growth of clinical \u003cem\u003eS. aureus\u003c/em\u003e (CSA) and standard \u003cem\u003eP. aeruginosa\u003c/em\u003e (SPA) with fewer significant antibacterial effects (55% and 56%). The effect of \u003cem\u003eArtemisia sieberi\u003c/em\u003e essential oil (ASEO) at different concentrations on the targeted bacterial growth is reported in Fig. 1b. ASEO mostly restricted the SSA growth (20%), while other results showed lower antibacterial effects (CSA: 42%; SPA: 38%; CPA: 43%). As shown in Fig. 1c, \u003cem\u003eLaurus nobilis\u003c/em\u003e essential oils (LNEO) significantly inhibited the growth of SSA (39%), SPA (35%), and CPA (38%). Moreover, LNEO decreased the activity of CSA with fewer effects (51%).\u003c/p\u003e\n\u003cp\u003eThe antibacterial effects of FGEO were observed in Fig. 1d. Totally, FGEO showed one of the best results in the present study and remarkably inhibited all spp. While the growth of CSA was decreased to 17%, the growth of SSA, SPA, and CPA was reduced to 39%, 36%, and 41%.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLippia citriodora\u003c/em\u003e essential oils (LCEO) showed significant inhibitory effects with increasing the concentration (Fig. 1e). The growth of all targeted bacteria was inhibited. LCEO at a concentration of 8.00 mg/mL reduced the growth of CPA and CSA to 44% and 39%. On the other hand, the growth restriction of standard spp. higher than in clinical specimens (SPA, 33%; SSA, 37%).\u003c/p\u003e\n\u003cp\u003eResults of bacterial inhibitory effects of \u003cem\u003eCymbopogon citratus\u003c/em\u003e essential oil (CIEO) are shown in Fig. 1f. Mostly, CIEO showed antibacterial activity on SPA (27%), but CIEO totally showed significant inhibitory effects (more than 50% bacterial growth inhibition) on all the other spp. (SSA, 30%; CSA, 32%; CPA, 38%).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisinfection activity of FGEO on trolley surface\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTable 2 shows the antibacterial effects of FGEO on the surface of trolley contaminated with standard and clinical \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e. FGEO significantly decreased the growth of bacterial agents in all samples. In addition, the antibacterial effects were positively related to the concentration and the time of exposure.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e\u003cstrong\u003e\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/strong\u003e antibacterial effects of FGEO on trolley surface contominated with S. S. aureus, C.S.aureus, S.P.aeroginosa, C.P.aeroginosa.\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"13.26530612244898%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"22.448979591836736%\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"23.46938775510204%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4000 mg/mL\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"22.448979591836736%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8000 mg/mL\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.606741573033707%\"\u003e\n \u003cp\u003eTime of exposure to FGEO (minutes)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.741573033707865%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.3707865168539324%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.606741573033707%\"\u003e\n \u003cp\u003eThe mean number of S.S. aureus colonies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.741573033707865%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.3707865168539324%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.606741573033707%\"\u003e\n \u003cp\u003eThe mean number of C.S. aureus colonies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.741573033707865%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.3707865168539324%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.606741573033707%\"\u003e\n \u003cp\u003eThe mean number of S.P. aeruginosa colonies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.741573033707865%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.3707865168539324%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.606741573033707%\"\u003e\n \u003cp\u003eThe mean number of C.P. aeruginosa colonies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.741573033707865%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.617977528089888%\"\u003e\n \u003cp\u003e10\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.3707865168539324%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.49438202247191%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: positive control, washed with normal saline; negative control, washed with alcohol; FGEO, \u003cem\u003eFerula Gummosa\u003c/em\u003e essential oil; S.S. aureus, Standard \u003cem\u003eStaphylococcus aureus\u003c/em\u003e; C.S. aureus, clinical \u003cem\u003eStaphylococcus aureus\u003c/em\u003e; S.P. aeruginosa, Standard \u003cem\u003epseudomonas aeruginosa\u003c/em\u003e; C.P. aeroginosa, clinical \u003cem\u003epseudomonas aeruginosa\u003c/em\u003e.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study aimed to evaluate the antibacterial effects of six EOs at the various concentrations on standard and clinical species of two common nosocomial pathogens (\u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e). In addition, we selected the most efficient antibacterial EO to investigate its antibacterial effects on nosocomial surfaces.\u003c/p\u003e\n\u003cp\u003eGenerally, \u003cem\u003eS. aureus\u003c/em\u003e is found approximately all over the environment, especially on the skin and nasal cavity. On the other hand, \u003cem\u003eS. aureus\u003c/em\u003e is one the most opportunistic pathogens that would result in several nosocomial infections\u0026nbsp;\u003csup\u003e14\u003c/sup\u003e. The results showed that ASEO had the best antibacterial effects on SSA among examined EOs. In the following, CCEO and CIEO represented acceptable effects, but the other EOs had the lower inhibitory activity on SSA.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eP. aeruginosa\u003c/em\u003e is another opportunistic pathogen that belongs to gram-negative strains. The genus \u003cem\u003ePseudomonas\u003c/em\u003e is available to survive in various difficult environments and causes life-threatening nosocomial infections. Totally, the experimented EO in the present study showed fewer antibacterial effects against \u003cem\u003eP. aeruginosa\u003c/em\u003e. The best findings were observed in the case of CIEO, but the other EOs showed significant inhibitory effects, except CCEO\u0026nbsp;\u003csup\u003e15\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe genus \u003cem\u003eArtemisia\u003c/em\u003e is a well-known medicinal plant that grows worldwide\u0026nbsp;\u003csup\u003e16\u003c/sup\u003e. \u003cem\u003eArtemisia sieberi\u003c/em\u003e is a special species of this large genus usually found in the northern regions of Iran and Saudi Arabia. Chemical analysis revealed that ASEO is mostly made up of \u003cem\u003eArtemisia\u0026nbsp;\u003c/em\u003eketone and 1,8-cineole. Moreover, Vahdani et al. reported that the main compounds of ASEO were piperitone, camphor, and 1, 8-cineole\u0026nbsp;\u003csup\u003e17\u003c/sup\u003e. On the other hand, a recent GC-MS indicated that Camphor, 1,8-cineole, and \u0026beta;-Thujone were the first major components of ASEO\u0026nbsp;\u003csup\u003e18\u003c/sup\u003e. Therefore, we can conclude that 1,8-cineole is an improved component in the chemical composition of ASEO and would be one of the major antibacterial agents of this EO. Different species of genus \u003cem\u003eArtemisia\u003c/em\u003e have encouraging antibacterial activities\u0026nbsp;\u003csup\u003e19\u003c/sup\u003e. Also, \u003cem\u003eA. sieberi\u003c/em\u003e products indicated great inhibitory effects on both Gram-negative and Gram-positive strains\u0026nbsp;\u003csup\u003e20\u003c/sup\u003e. In 2013, Hedayati rad et al. conducted a novel Pullulan film incorporated with ASEO to evaluate its protective effects against different bacterial pathogens. This study revealed that ASEO at low concentrations (5%) has no inhibitory effects on bacterial growth, but with increasing the concentration (at least to 10%) the antibacterial effects would develop. The highest inhibitory zone (18.52 \u0026plusmn; 0.73) was observed at 35%\u0026nbsp;\u003csup\u003e21\u003c/sup\u003e. Pure ASEO made large inhibitory zones against \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e in Muller Hinton agar plates. Besides, incorporating basal seed gum films with ASEO resulted in a big inhibitory zone (23.2 \u0026plusmn; 0.7 mm) on plates infected with \u003cem\u003eS. aureus\u003c/em\u003e and a smaller inhibitory zone (10.2 \u0026plusmn; 0.6 mm) in the case of \u003cem\u003eP. aeruginosa\u003c/em\u003e \u003csup\u003e22\u003c/sup\u003e. Supporting the results of the present study, others also reported that ASEO represented valuable MIC (10 \u0026micro;L) and minimal bactericidal concentration (MBC) (20 \u0026micro;L) against \u003cem\u003eS. aureus\u003c/em\u003e \u003csup\u003e17\u003c/sup\u003e. Moreover, recent studies on synergic antibacterial effects of ASEO combined with different routine antibiotics demonstrated that ASEO would increase the inhibitory effects of ampicillin, oxacillin, cephalothin, piperacillin, and vancomycin on \u003cem\u003eS. aureus\u003c/em\u003e. On the other hand, the combination of various antibiotics, such as piperacillin, gentamicin, ampicillin, and erythromycin with ASEO indicated significant synergic effects on the growth of \u003cem\u003eP. aeruginosa\u003c/em\u003e \u003csup\u003e23\u003c/sup\u003e. Therefore, using pure ASEO and its combination with different products would provide novel antibacterial materials that represent higher inhibitory effects and decrease the difficulties of previous routine antibiotics.\u003c/p\u003e\n\u003cp\u003eChemical composition analysis indicated that CCEO mostly contains 1-phenylpropanol and gamma-Terpinene\u0026nbsp;\u003csup\u003e24\u003c/sup\u003e. Besides, Hajlaoui et al. reported that CCEO mainly comprised cumin aldehyde and gamma-terpinene. On the other hand, the first evaluations revealed that Isobutyl isobutyrate, a-Thujene, and a-Pinene are the most common compositions in CCEO. CCEO is believed to have a protective role in food production and storage industries\u0026nbsp;\u003csup\u003e25\u003c/sup\u003e. Further studies demonstrated that the most protective CCEO concentrations will appear at 0.03% that prevented the \u003cem\u003eS. aureus\u003c/em\u003e growth in feta cheese and infected hamburgers\u0026nbsp;\u003csup\u003e24\u003c/sup\u003e. CCEO demonstrated promising MBC and MIC on SSA (2.5 to 5 \u0026micro;g/mL, and 1.25 to 5 \u0026micro;g/mL). Moreover, this study showed that CCEO had the same antibacterial effect on SSA as vancomycin, a highly used antibiotic with several adverse effects\u0026nbsp;\u003csup\u003e26\u003c/sup\u003e. On the other hand, some other studies supported our results. Saee et al. reported that the extract and EO from \u003cem\u003eC. cyminum\u003c/em\u003e significantly diminished the CPA with higher efficiency than several antibiotics such as gentamicin, amoxicillin, and clotrimazole\u0026nbsp;\u003csup\u003e27\u003c/sup\u003e. Therefore, utilizing CCEO in preventing and treating procedures would decrease the limitations of routine antibiotics in the future, but the controversies of its antibacterial effects on gram-negative strains remained for further studies.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eC. citratus\u003c/em\u003e has a valuable protecting usage in food industries to restrict the activity of common food-borne pathogens. Utilizing CIEO in the preservation of cooked beef patties would save its quality and prevent the growth of pathogenic bacterial strains\u0026nbsp;\u003csup\u003e28\u003c/sup\u003e. The evaluations showed that neral, geranial, limonene, and geranyl acetate are the major components of CIEO. \u003cem\u003eC. citratus\u003c/em\u003e products mainly prevent the growth of fungal (\u003cem\u003eCandida\u003c/em\u003e strains) and Gram-positive bacterial species, such as \u003cem\u003eS. aureus\u003c/em\u003e, biofilms. Based on the last in vitro trials, the CIEO provides the same antibacterial effects on \u003cem\u003eS. aureus\u003c/em\u003e as vancomycin and would be a favorable product to prevent the growth of nosocomial pathogens with fewer adverse effects in the future\u0026nbsp;\u003csup\u003e29\u003c/sup\u003e. In addition, a recent study on EO from the leaves of \u003cem\u003eC. citratus\u003c/em\u003e indicated a bigger zone of inhibition in the case of \u003cem\u003eS. aureus\u003c/em\u003e (32.0 \u0026plusmn; 0.75 mm) and no antibacterial effects on \u003cem\u003eP. aeruginosa\u003c/em\u003e. Nevertheless, the other studies showed that \u003cem\u003eC. citratus\u003c/em\u003e extracts would represent more antibacterial effects on gram-negative strains such as \u003cem\u003eP. aeruginosa\u003c/em\u003e. Subramanian et al. reported that the leaf and root extracts resulted in a very large zone of inhibitions (\u0026gt; 30 mm), and would be encouraging herbal products against these pathogenic gram-negative strains\u0026nbsp;\u003csup\u003e30\u003c/sup\u003e. Moreover, the results of our study represented that CIEO mainly decreased the growth of SPA more than the other targeted strains. Therefore, further studies are required to evaluate these controversial results.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eL. citriodora\u003c/em\u003e and hundreds of other species in the \u003cem\u003eLippia\u003c/em\u003e genus are well-known ancient plants utilized for several decades, especially in Western regions of the world. GC-MS analysis reported that neral and geranial are the major compounds of experimented LCEO\u0026nbsp;\u003csup\u003e31\u003c/sup\u003e. In addition, other studies added terpenic alcohols, monoterpenic hydrocarbons (dl-limonene), and \u0026alpha;-curcumene to previously reported chemical components. Several studies reported various medicinal properties, such as sedative, diuretic, and antispasmodic use in recent years. Moreover, the recent papers introduced LCEO as a moderate antibacterial product\u0026nbsp;\u003csup\u003e32\u003c/sup\u003e. Although pure LCEO provided an acceptable zone of inhibition on sterile Whatman paper discs infected with gram-positive (\u003cem\u003eS. aureus\u003c/em\u003e) and gram-negative (\u003cem\u003eE. coli\u003c/em\u003e) strains, this EO mainly affects the gram-negative strains more than gram-positive species. On the other hand, some other studies represented controversial results. Evaluating the effects of LCEO on different pathogens showed that this EO has no significant inhibitory or bactericidal effects on \u003cem\u003eP. aeruginosa\u003c/em\u003e and the MIC and MBC are higher in the case of \u003cem\u003eE. coli\u003c/em\u003e than \u003cem\u003eS. aureus\u003c/em\u003e \u003csup\u003e33\u003c/sup\u003e. Therefore, the present study and all other studies introduced LCEO as an encouraging antibacterial product, especially for gram-positive strain, but further studies are required to put an end to the controversial results in the case of \u003cem\u003eP. aeruginosa\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLaurus nobilis\u003c/em\u003e is another aromatic plant with antimicrobial and flavor properties in food industries. According to the high requirement of enough water and humidity, \u003cem\u003eL. nobilis\u003c/em\u003e usually grows in warmer climate places such as the south of the Mediterranean region. Methyl eugenol, 1,8-cineole, and \u0026alpha;-terpinyl acetate are the major compounds of LNEO. LNEO has significant inhibitory effects on all gram-positive and -negative strains, like \u003cem\u003eP. aeruginosa\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eE. coli\u003c/em\u003e. The measurements of LNEO inhibition zone showed higher antibacterial effects than tetracycline (8 mg/mL), especially in the case of gram-positive strains. The evaluations revealed that LNEO has high antibacterial effects on gram-positive strains such as \u003cem\u003eS. aureus\u003c/em\u003e (inhibition zone = 13 mm), \u003cem\u003eS. intermedius\u003c/em\u003e, and \u003cem\u003eK. pneumoniae\u003c/em\u003e. Also, LNEO significantly reduces the biofilm activity with acceptable MIC, ranged from 0.3 to 96.4 mg/mL\u0026nbsp;\u003csup\u003e34\u003c/sup\u003e. Further comparisons showed that gram-positive strains are more susceptible to LNEO than \u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e. The recent evaluations demonstrated that minimal inhibitory activities of LNEO would be represented at 0.25 mg/mL. On the other hand, LNEO showed remarkable protective effects against common foodborne pathogens, such as \u003cem\u003eE. coli\u003c/em\u003e \u003csup\u003e35\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe aromatic herbal families, such as \u003cem\u003eApiaceae\u003c/em\u003e, are common plants that grow in different regions of Iran. The genus of \u003cem\u003eFerula\u003c/em\u003e is a medicinal herb mostly found in EMERO and north Africa floras\u0026nbsp;\u003csup\u003e36\u003c/sup\u003e. Several studies revealed different protective effects of \u003cem\u003eF. gummosa\u003c/em\u003e against infection, inflammatory and spasmodic processes. EO from \u003cem\u003eF. gummosa\u003c/em\u003e seeds mostly showed strong antibacterial effects on gram-positive strains and slight inhibitory activity against gram-negative strains, such as \u003cem\u003eP. aeruginosa\u003c/em\u003e. Abbaszadegan et al. reported that \u0026beta;-pinene made up more than 50% of the chemical composition of FGEO. Moreover, FGEO at 50 \u0026micro;g/mL resulted in a big inhibitory zone (22.5 \u0026plusmn; 0.82 mm) on plates infected with \u003cem\u003eS. aureus\u003c/em\u003e \u003csup\u003e37\u003c/sup\u003e. Considering the antibacterial effects on both S. aureus and P. aeruginosa, our findings showed that FGEO is the most effective agent among the evaluated EOs in the present study. As a result, we selected FGEO to investigate its disinfection activity on the surface of emergency hospital.\u003c/p\u003e\n\u003cp\u003eIn the last two decades, several studies have attempted to use EO-based disinfectants instead of conventional disinfectants. Sharma et al. indicated that a novel green disinfectant, named Neutral Biodegradable Disinfectant (BND) would appropriately clear the contaminated surfaces. Their findings confirmed that phenol-based disinfectants and NBD products disinfect environmental microorganisms equally\u0026nbsp;\u003csup\u003e38\u003c/sup\u003e. As a result, given that natural products provide much less resistance and reduce the population of microorganisms evenly, many of these products are expected to replace conventional phenol-based disinfectants in the coming years\u0026nbsp;\u003csup\u003e39\u003c/sup\u003e. Increasing bacterial resistance to conventional disinfectants resulted in a great challenge in different healthcare services, especially in dental procedures\u0026nbsp;\u003csup\u003e40\u003c/sup\u003e. In recent years, several observations indicated that green disinfectants would become one of the main agents to provide streel dental practices. In 2019, Nausheen et al. confirmed that aloe vera gel at high concentrations (90%) is a safe disinfectant and shows less poor outcomes than conventional chemical products\u0026nbsp;\u003csup\u003e41\u003c/sup\u003e. In addition, our findings supported the previous results. FGEO relatively could decrease the growth of both bacterial agents on the trolley surface. Although the increasing concentration of FGEO developed its antibacterial activity, a significant population of bacterial colonies grew on the evaluated samples of our study. Therefore, the application of FGEO-based disinfectants would not be able to completely clear nosocomial surfaces, similar to conventional and novel green disinfectants\u0026nbsp;\u003csup\u003e42\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe present study has several strengths and limitations. The main strength of our study was the comparison of the antibacterial effects of above-mentioned EOs on clinical and standard samples of two major gram-positive and gram-negative strains. Based on our expanded literature review on previous studies, we mainly hypothesized that the gram-negative strains and clinical samples would represent more resistance against different antibacterial EOs. Generally, the findings supported these hypotheses with little controversy (especially in the case of CIEO and LNEO). Moreover, we faced some conflicts in comparison with previous studies. On the other hand, we just reported the inhibition activity of EOs on bacterial growth in percent. Therefore, further studies with more targeted pathogens and expand measurements would be required to collect more sufficient data and resolve observed controversies.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eBacterial species\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn this study, we obtained standard bacterial species from the Microbiology laboratory of Pasteur Institute of Tehran (Standard species of \u003cem\u003eS. aureus\u003c/em\u003e, ATCC 25923; Standard species of \u003cem\u003eP. aeruginosa\u003c/em\u003e, ATCC 27853). On the other hand, we used clinical isolated Methicillin-resistant \u003cem\u003eS. aureus\u003c/em\u003e (MRSA) and \u003cem\u003eP. aeruginosa\u003c/em\u003e Metallo-beta-lactamase (MBL)-producing from wound infection from previous studies of our researches. Also, we utilized molecular method (PCR test) and the routine culture media (the disk-diffusion method and minimal inhibitory concentration (MIC) test) tests to identify phenotypically and genotypically MBL-producing \u003cem\u003eP. aeruginosa\u003c/em\u003e and MRSA \u003csup\u003e43\u003c/sup\u003e.\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003c/span\u003eWe obtained all EOs from Tabib Daroo Company (a pharmaceutical company in Kashan). Herbarium specifications of each plant are shown in the Table 3.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3.\u003c/strong\u003e Herbarium specifications of applied plants\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003eScientific names (Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003ePart used\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eHerbarium code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eCollector\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003ePlace of collection\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003e\u003cem\u003eCuminum \u0026nbsp; \u0026nbsp; cyminum\u003c/em\u003e (Cuminum Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eSeed\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eIAUM-\u003c/p\u003e\n \u003cp\u003e211\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eDr. A. Ziaeei\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eBajestan, \u0026nbsp;khorasan, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003e\u003cem\u003eArtemisia sieberi\u003c/em\u003e (Asteraceae Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eLeave\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eHUMZ-8629\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eDr. A. Taheri\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eGhaemshahr, Mazandaran, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003e\u003cem\u003eLaurus nobilis\u003c/em\u003e (Lauraceae Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eLeave\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eIAUGH-1409\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eDr.\u0026nbsp;E. Bahrani\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eTehran,\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eTehran, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003e\u003cem\u003eFerula gummosa\u003c/em\u003e (Apiaceae Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eAll\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eUQH-1530\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eDr. A.\u003c/p\u003e\n \u003cp\u003eKhani\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eArak,\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eMarkazi, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003e\u003cem\u003eLippia citriodora\u003c/em\u003e (Verbenaceae Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eLeave\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eKF-1513\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eDr. Mehrabani\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eSirch,\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eKerman, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"7.532051282051282%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.16025641025641%\"\u003e\n \u003cp\u003eCymbopogon citratus (Poaceae\u0026nbsp;Family)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eLeave\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.826923076923077%\"\u003e\n \u003cp\u003eHUI-TB400CI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.26923076923077%\"\u003e\n \u003cp\u003eA. Mazaheri\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.71153846153846%\"\u003e\n \u003cp\u003eKashan,\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eIsfahan, Iran\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eBacterial culture\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe used Muller Hinton broth, containing 0.5% DMSO for bacterial growth (Merck Chemicals, Germany) \u003csup\u003e44\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGas Chromatography/Mass Spectrometry analysis\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eGC-MS analysis was deployed to identify the compounds of \u003cs\u003ethe\u003c/s\u003e selected herbal EOs. We utilized a 7890A Network GC system alongside a 5975A massive detector (Agilent Technologies, Santa Clara, CA, USA). The compounds of EOs were separated via fused columns of HP-5MS silica (length, 30 meters; internal diameters, 250 µM; film thickness, 25 µM). The temperature was set first at 50°C, increased gradually to 200°C (5°C per minute), and remained at this temperature for 1 minute. Then, we elevated the temperature up to 250°C (10°C per minute) and again increased to 300°C with higher rate (25°C per minute). On the other hand, helium was utilized to carry the ingredients. At last, we analyzed the components based on our previously described method \u003csup\u003e45\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetermination of the antibacterial activity of Eos\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe utilized a 96-well plate to provide a proper situation for evaluation of antibacterial effects of EOs against purposed bacterial species based on standard protocols \u003csup\u003e46\u003c/sup\u003e. We achieved 1.5 × 10\u003csup\u003e8\u003c/sup\u003e CFU/mL of bacterial colonies to pass the 0.5 McFarland turbidity. Then 20-µL suspension was added to all wells. After preparation of bacterial agents, we dissolved EOs in Muller Hinton Broth to provide a standard dilution series (8.00, 4.00, 2.00, 1.00, 0.50, 0.25, and 0.13 mg. mL-1). After described dilution, we added 80 µL of broth suspension to all wells and incubated in standard set (24h incubation at 37°C). The turbidity evaluation was performed at 630 nm via plate reader (Synergy HTX Multi-Mode Reader, USA) and the bacterial growth was assessed through Equation 1.\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMeasurement of disinfection properties of \u003cem\u003eFerula Gummosa\u003c/em\u003e essential oil (FGEO)\u0026nbsp;\u003c/strong\u003eFGEO were prepared in two different concentrations (4000 and 8000 µg) to measure its disinfection activity on 72 samples (3 × 3 cm\u003csup\u003e2\u003c/sup\u003e) from different emergency trolley steel surfaces (three times). First, we cleaned the samples using 70% ethyl alcohol and sterile cotton soaked in sterile water (3 times, each time from the center to the outside). Then, all samples were contaminated with bacterial agents by streel swab and they were allowed to drain at room temperature. The bacterial strains were cultured in Muller Hinton Agar (MHA) and suspension of 0.5 McFarland for 24 to 48 hours (approximately 1.5 × 10\u003csup\u003e8\u003c/sup\u003e bacterial colonies). Finally, all surfaces were divided into four groups (alcohol as negative control, untreated surface as positive control, 4000 FGEO, and 8000 FGEO). Each group divided into four subgroups of bacterial species (\u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e). The final samples were gathered after 1, 3, and 5 minutes. The samples were carried to the laboratory in a freezer to prevent probable growth. Each sample vortexed for 1 minute and 100 µL of the sample was cultured on blood agar. The broths were kept in an incubator for 24 hours at 37℃. Finally, the bacterial colonies of each sample were counted and the mean was reported \u003csup\u003e13\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical methods\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe carried out antibacterial tests in triplicates. SPSS software (Version 22, SPSS Inc, USA) was utilized to measure the means ± standard deviations and draw charts. We deployed a regression model (performed in CalcuSyn software, Free version, BIOSOFT, UK) to calculate IC50 of all Eos; then, we compared all IC50 of the Eos together via one-way ANOVA using SPSS software (Version 22, SPSS Inc, USA) independent sample t-test. The confidence interval of this study was set at 95% (CI 95%).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study indicated that all experimented EOs has encouraging antibacterial effects against targeted pathogens. Some products, such as CCEO, ASEO, and FGEO represented higher inhibitory effects on \u003cem\u003eS. aureus\u003c/em\u003e and the latter, including CIEO and LNEO, had better effects on \u003cem\u003eP. aeruginosa\u003c/em\u003e. Therefore, further studies are required to evaluate the antibacterial effects of the combination of these EOs and utilize novel techniques, such as nanoparticles to develop more efficient natural products and prevent life-threatening nosocomial infections. Since essential oils can have synergistic or antagonistic effects on each other, it is suggested to investigate the effects of the combination of essential oils. Nonetheless, it will be considered in our additional studies in the future.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors wish to acknowledge the support of Fasa University of Medical Sciences for facilitating this research.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed to the study conception and design.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by a grant from the\u0026nbsp;Fasa University of Medical Sciences.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors confirm that the data supporting the findings of this study are available within the article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll other authors have no conflicts of interest to declare.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eSuetens, C. \u003cem\u003eet al.\u003c/em\u003e Prevalence of healthcare-associated infections, estimated incidence and composite antimicrobial resistance index in acute care hospitals and long-term care facilities: results from two European point prevalence surveys, 2016 to 2017. 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Nanomedicine Research Journal \u003cb\u003e3\u003c/b\u003e, 37\u0026ndash;43 (2018).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Essential oil, Antibacterial activity, Microdilution, Disinfectant","lastPublishedDoi":"10.21203/rs.3.rs-2467112/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2467112/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe component and antibacterial effects of six essential oils )EOs(, including \u003cem\u003eCuminum cyminum\u003c/em\u003e (CCEO), \u003cem\u003eArtemisia sieberi\u003c/em\u003e (ASEO), \u003cem\u003eLaurus nobilis\u003c/em\u003e (LNEO), \u003cem\u003eFerula Gummosa essential\u003c/em\u003e (FGEO), \u003cem\u003eLippia citriodora\u003c/em\u003e (LCEO), \u003cem\u003eCymbopogon citratus\u003c/em\u003e (CIEO) were measured by GC-MS and 96-well micro-plates (IC50), against \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e. Then, the antibacterial effects of FGEO, the most efficient EO, were examined on the trolley surface in hospital for 1, 3, 5 and 10 min intervals. While CCEO, ASEO, and FGEO have the highest effects on the growth of \u003cem\u003eS. aureus\u003c/em\u003e, CIEO and LNEO showed the best outcomes on \u003cem\u003eP. aeruginosa\u003c/em\u003e. In addition, our finding showed that FGEO relatively decreases the growth of \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eP. aeruginosa\u003c/em\u003e on the trolley surface (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In addition, FGEO, one of the best disinfectant agents of the present study, showed relatively antibacterial effects on the surface of the trolley.\u003c/p\u003e","manuscriptTitle":"Antibacterial Activity of Some Iranian Plant Essential Oils as Disinfectant Agents on Surfaces Contaminated with Staphylococcus Aureus and Pseudomonas Aeruginosa","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-01-31 15:30:44","doi":"10.21203/rs.3.rs-2467112/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"07113207-c44a-4300-a695-da95bca99ee1","owner":[],"postedDate":"January 31st, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":18757339,"name":"Biological sciences/Biotechnology"},{"id":18757340,"name":"Biological sciences/Chemical biology"},{"id":18757341,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2023-02-25T09:59:54+00:00","versionOfRecord":[],"versionCreatedAt":"2023-01-31 15:30:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2467112","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2467112","identity":"rs-2467112","version":["v1"]},"buildId":"-HB7Z8yhvgn0wM9Nzuekk","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}