Effect of Matricaria aurea Essential Oils on Biofilm Development, Virulence Factors and Quorum Sensing-Dependent Genes of Pseudomonas aeruginosa.

The emergence of drug-resistant microorganisms presents a substantial global public health threat. The increase in pathogens resistant to commonly prescribed antibiotics underscores the urgent requirement to explore alternative treatment strategies. This study adopts a novel approach by harnessing natural resources, specifically essential oils (EO), to combat bacterial pathogenicity. The primary aim of this research was to analyze the chemical composition of the aerial part of the Matricaria aurea (M. aureas) EO and evaluate its potential for inhibiting quorum sensing (QS) and disrupting biofilm formation in Pseudomonas aeruginosa (P. aeruginosa). The gas chromatography-mass spectrometry (GCMS) analysis unveiled that α-bisabolol oxide A constituted the predominant portion, comprising 64.8% of the total, with ß-bisabolene at 6.3% and α-farnesene at 4.8% following closely behind. The antibiofilm efficacy was observed at concentrations of 0.3, 0.15, and 0.08 mg/mL, demonstrating negligible effects on cell viability. Furthermore, the EO from M. aurea effectively inhibited the formation of P. aeruginosa biofilms by diminishing aggregation, hydrophobicity, and swarming motility. Significantly, the EO treatment resulted in a conspicuous decrease in the production of pyocyanin, rhamnolipid, and extracellular polymeric substances (EPS), along with a reduction in the enzymatic activity of protease and chitinase. The EO effectively hindered QS by disrupting QS mechanisms, resulting in a marked decline in the secretion of N-Acyl homoserine lactone (AHL) molecules and the expression of phazA1 and aprA genes. This investigation offers compelling evidence supporting the potential of M. aurea EO as a promising therapeutic candidate for addressing infectious diseases induced by biofilm formation.

Memory Enhancing and Neurogenesis Activity of Honey Bee Venom in the Symptoms of Amnesia: Using Rats with Amnesia-like Alzheimer's Disease as a Model.

BACKGROUND/OBJECTIVE: Alzheimer's disease (AD) is mainly characterized by amnesia that affects millions of people worldwide. This study aims to explore the effectiveness capacities of bee venom (BV) for the enhancement of the memory process in a rat model with amnesia-like AD. METHODS: The study protocol contains two successive phases, nootropic and therapeutic, in which two BV doses (D1; 0.25 and D2: 0.5 mg/kg i.p.) were used. In the nootropic phase, treatment groups were compared statistically with a normal group. Meanwhile, in the therapeutic phase, BV was administered to scopolamine (1mg/kg) to induce amnesia-like AD in a rat model in which therapeutic groups were compared with a positive group (donepezil; 1mg/kg i.p.). Behavioral analysis was performed after each phase by Working Memory (WM) and Long-Term Memory (LTM) assessments using radial arm maze (RAM) and passive avoidance tests (PAT). Neurogenic factors; Brain-derived neurotrophic factor (BDNF), and Doublecortin (DCX) were measured in plasma using ELISA and Immunohistochemistry analysis of hippocampal tissues, respectively. RESULTS: During the nootropic phase, treatment groups demonstrated a significant (P < 0.05) reduction in RAM latency times, spatial WM errors, and spatial reference errors compared with the normal group. In addition, the PA test revealed a significant (P < 0.05) enhancement of LTM after 72 hours in both treatment groups; D1 and D2. In the therapeutic phase, treatment groups reflected a significant (P < 0.05) potent enhancement in the memory process compared with the positive group; less spatial WM errors, spatial reference errors, and latency time during the RAM test, and more latency time after 72 hours in the light room. Moreover, results presented a marked increase in the plasma level of BDNF, as well as increased hippocampal DCX-positive data in the sub-granular zone within the D1 and D2 groups compared with the negative group (P < 0.05) in a dose-dependent manner. CONCLUSION: This study revealed that injecting BV enhances and increases the performance of both WM and LTM. Conclusively, BV has a potential nootropic and therapeutic activity that enhances hippocampal growth and plasticity, which in turn improves WM and LTM. Given that this research was conducted using scopolamine-induced amnesia-like AD in rats, it suggests that BV has a potential therapeutic activity for the enhancement of memory in AD patients in a dose-dependent manner but further investigations are needed.

Synergistic Effects of AgNPs and Biochar: A Potential Combination for Combating Lung Cancer and Pathogenic Bacteria.

The synthesis of reliable biological nanomaterials is a crucial area of study in nanotechnology. In this study, Emericella dentata was employed for the biosynthesis of AgNPs, which were then combined with synthesized biochar, a porous structure created through biomass pyrolysis. The synergistic effects of AgNPs and biochar were evaluated through the assessment of pro-inflammatory cytokines, anti-apoptotic gene expression, and antibacterial activity. Solid biosynthesized AgNPs were evaluated by XRD and SEM, with SEM images revealing that most of the AgNPs ranged from 10 to 80 nm, with over 70% being less than 40 nm. FTIR analysis indicated the presence of stabilizing and reducing functional groups in the AgNPs. The nanoemulsion's zeta potential, hydrodynamic diameter, and particle distribution index were found to be -19.6 mV, 37.62 nm, and 0.231, respectively. Biochar, on the other hand, did not have any antibacterial effects on the tested bacterial species. However, when combined with AgNPs, its antibacterial efficacy against all bacterial species was significantly enhanced. Furthermore, the combined material significantly reduced the expression of anti-apoptotic genes and pro-inflammatory cytokines compared to individual treatments. This study suggests that low-dose AgNPs coupled with biochar could be a more effective method to combat lung cancer epithelial cells and pathogenic bacteria compared to either substance alone.

Chemical Composition and Quorum Sensing Inhibitory Effect of Nepeta curviflora Methanolic Extract against ESBL Pseudomonas aeruginosa.

Objectives: Bacterial biofilm is regarded as a significant threat to the production of safe food and the arise of antibiotic-resistant bacteria. The objective of this investigation is to evaluate the quorum sensing inhibitory effect of Nepeta curviflora methanolic extract. Methods: The effectiveness of the leaves at sub-inhibitory concentrations of 2.5, 1.25, and 0.6 mg/mL on the virulence factors and biofilm formation of P. aeruginosa was evaluated. The effect of N. curviflora methanolic extract on the virulence factors of P. aeruginosa, including pyocyanin, rhamnolipid, protease, and chitinase, was evaluated. Other tests including the crystal violet assay, scanning electron microscopy (SEM), swarming motility, aggregation ability, hydrophobicity and exopolysaccharide production were conducted to assess the effect of the extract on the formation of biofilm. Insight into the mode of anti-quorum sensing action was evaluated by examining the effect of the extract on the activity of N-Acyl homoserine lactone (AHL) and the expression of pslA and pelA genes. Results: The results showed a significant attenuation in the production of pyocyanin and rhamnolipid and in the activities of protease and chitinase enzymes at 2.5 and 1.25 mg/mL. In addition, N. curviflora methanolic extract significantly inhibited the formation of P. aeruginosa biofilm by decreasing aggregation, hydrophobicity, and swarming motility as well as the production of exopolysaccharide (EPS). A significant reduction in AHL secretion and pslA gene expression was observed, indicating that the extract inhibited quorum sensing by disrupting the quorum-sensing systems. The quorum-sensing inhibitory effect of N. curviflora extract appears to be attributed to the presence of kaempferol, quercetin, salicylic acid, rutin, and rosmarinic acid, as indicated by LCMS analysis. Conclusion: The results of the present study provide insight into the potential of developing anti-quorum sensing agents using the extract and the identified compounds to treat infections resulting from quorum sensing-mediated bacterial pathogenesis.

Antioxidant and Antihyperglycemic Effects of Ephedra foeminea Aqueous Extract in Streptozotocin-Induced Diabetic Rats.

BACKGROUND: Ephedra foeminea is known in Jordan as Alanda and traditionally. It is used to treat respiratory symptoms such as asthma and skin rashes as an infusion in boiling water. The purpose of this study was to determine the antidiabetic property of Ephedra foeminea aqueous extract in streptozotocin-induced diabetic rats. METHODS: The aqueous extract of Ephedra foeminea plant was used to determine the potential of its efficacy in the treatment of diabetes, and this extract was tested on diabetic rats as a model. The chemical composition of Ephedra foeminea aqueous extract was determined using liquid chromatography-mass spectrometry (LC-MS). Antioxidant activity was assessed using two classical assays (ABTS and DPPH). RESULTS: The most abundant compounds in the Ephedra foeminea extract were limonene (6.3%), kaempferol (6.2%), stearic acid (5.9%), ß-sitosterol (5.5%), thiamine (4.1%), riboflavin (3.1%), naringenin (2.8%), kaempferol-3-rhamnoside (2.3%), quercetin (2.2%), and ferulic acid (2.0%). The antioxidant activity of Ephedra foeminea aqueous extract was remarkable, as evidenced by radical scavenging capacities of 12.28 mg Trolox/g in ABTS and 72.8 mg GAE/g in DPPH. In comparison to control, induced diabetic rats treated with Ephedra foeminea extract showed significant improvement in blood glucose levels, lipid profile, liver, and kidney functions. Interleukin 1 and glutathione peroxidase levels in the spleen, pancreas, kidney, and liver of induced diabetic rats treated with Ephedra foeminea extract were significantly lower than in untreated diabetic rats. CONCLUSIONS: Ephedra foeminea aqueous extract appears to protect diabetic rats against oxidative stress and improve blood parameters. In addition, it has antioxidant properties that might be very beneficial medicinally.

Formononetin Ameliorates Renal Dysfunction, Oxidative Stress, Inflammation, and Apoptosis and Upregulates Nrf2/HO-1 Signaling in a Rat Model of Gentamicin-Induced Nephrotoxicity.

Gentamicin (GEN) is a bactericidal aminoglycoside known to cause nephrotoxicity. Formononetin (FN) is a potent flavonoid that exhibits numerous promising pharmacological activities. In this study, we have assessed the nephroprotective efficacy of FN against GEN-induced renal injury in rats. Rats were orally administered with FN (60 mg/kg/day, for 2 weeks) and were co-treated with intraperitoneal (i.p.) injection of GEN (100 mg/kg/day) during the days 8-14. GEN-treated rats demonstrated increased urea and creatinine levels in serum associated with marked histopathological changes in the kidney. Malondialdehyde (MDA) and protein carbonyl contents were elevated, whereas glutathione concentration and catalase and superoxide dismutase activities were lowered in GEN-administered rats. The FN largely prevented tissue damage, attenuated renal function, reduced MDA and protein carbonyl, and enhanced antioxidant capacity in the kidney of GEN-administrated animals. The kidney of GEN-treated rats demonstrated elevated Bax and caspase-3 protein expression, accompanied by lowered Bcl-2 protein expression, an effect that FN attenuated. Moreover, FN treatment caused upregulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) expression in renal tissue of GEN-intoxicated animals. Collectively, FN protects against GEN-caused renal damage via exhibiting antioxidant, anti-inflammatory, and antiapoptotic activities and augmenting Nrf2 signaling, suggesting FN as a promising agent for preventing drug-induced organ damage.

Ruta graveolens, Peganum harmala, and Citrullus colocynthis methanolic extracts have in vitro protoscolocidal effects and act against bacteria isolated from echinococcal hydatid cyst fluid.

Echinococcosis is a common and endemic disease that affects both humans and animals. In this study, the in vitro activities of methanolic extracts of Ruta graveolens, Peganum harmala aerial parts, and Citrullus colocynthis seeds against protoscolosis and isolated bacterial strains from hydatid cysts were assessed using disc diffusion methods and Minimum Inhibitory Concentration (MIC). The chemical composition of three methanolic extracts was studied using LC-MS. After 3 h of exposure to 40 mg/mL R. graveolens extract, a tenfold protoscolocidal effect was seen when compared to the convintional medication (ABZ) for the same duration (P < 0.05). The bacteria listed below were isolated from hydatid cyst fluid collected from a variety of sick locations, including the lung and liver. Micrococcus spp., E. coli, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter amnigenus, Pseudomonas aeruginosa, Staphylococcus xylosus, and Achromobacter xylosoxidans are among the bacteria that have been identified. The most effective extract was R. graveolens, followed by P. harmala and C. colocynthis, according to the results of antibacterial activity using the disc diffusion method. R. graveolens extract had the lowest MIC values (less than 2 mg/mL) against all microorganisms tested. This shows that the R. graveolens extract has additional properties, such as the ability to be both scolocidal and bactericidal. Because these bacteria are among the most prevalent pathogenic bacteria that increase the risk of secondary infection during hydatid cysts, the results of inhibitory zones and MICs of the R. graveolens methanol extract are considered highly promising.

Effectiveness of Anodal otDCS Following with Anodal tDCS Rather than tDCS Alone for Increasing of Relative Power of Intrinsic Matched EEG Bands in Rat Brains.

BACKGROUND: This study sought to determine whether (1) evidence is available of interactions between anodal tDCS and oscillated tDCS stimulation patterns to increase the power of endogenous brain oscillations and (2) the frequency matching the applied anodal otDCS's frequency and the brain's dominant intrinsic frequency influence power shifting during stimulation pattern sessions by both anodal DCS and anodal oscillated DCS. METHOD: Rats received different anodal tDCS and otDCS stimulation patterns using 8.5 Hz and 13 Hz state-related dominant intrinsic frequencies of anodal otDCS. The rats were divided into groups with specific stimulation patterns: group A: tDCS-otDCS (8.5 Hz)-otDCS (13 Hz); group B: otDCS (8.5 Hz)-tDCS-otDCS (13 Hz); group C: otDCS (13 Hz)-tDCS-otDCS (8.5 Hz). Acute relative power changes (i.e., following 10 min stimulation sessions) in six frequency bands-delta (1.5-4 Hz), theta (4-7 Hz), alpha-1 (7-10 Hz), alpha-2 (10-12 Hz), beta-1 (12-15 Hz) and beta-2 (15-20 Hz)-were compared using three factors and repeated ANOVA measurement. RESULTS: For each stimulation, tDCS increased theta power band and, above bands alpha and beta, a drop in delta power was observed. Anodal otDCS had a mild increasing power effect in both matched intrinsic and delta bands. In group pattern stimulations, increased power of endogenous frequencies matched exogenous otDCS frequencies-8.5 Hz or 13 Hz-with more potent effects in upper bands. The power was markedly more potent with the otDCS-tDCS stimulation pattern than the tDCS-otDCS pattern. SIGNIFICANCE: The findings suggest that the otDCS-tDCS pattern stimulation increased the power in matched intrinsic oscillations and, significantly, in the above bands in an ascending order. We provide evidence for the successful corporation between otDCS (as frequency-matched guidance) and tDCS (as a power generator) rather than tDCS alone when stimulating a desired brain intrinsic band (herein, tES specificity).

Silver nanoparticles biosynthesis using an airborne fungal isolate, Aspergillus flavus: optimization, characterization and antibacterial activity.

Background and Objectives: Nanoscience is one of the most important branches of modern science, which deals with the knowledge, structure, and properties of nanoparticles. This study aimed to investigate the ability of an airborne fungus (Aspergillus flavus) to synthesize silver nanoparticles (AgNPs) and to test the antibacterial activity of the synthesized AgNPs. Materials and Methods: The confirmation of AgNPs synthesis and the characterization of their properties were done using UV-Vis spectrophotometer, Zeta potential, Zeta sizer, FT-IR, and XRD analyses. The antibacterial activity was determined using broth microdilution method. Results: The findings showed that the average diameter of the resultant AgNPs was 474.2 nm with a PDI value of 0.27, and the zeta potential was -33.8 mV. Transmission electron microscopy (TEM) revealed that the AgNPs were regular and spherical in shape. TEM micrographs demonstrated that the AgNPs were smaller than those that were observed by DLS examination because the drying process resulted in particle shrinkage. The average size of AgNPs were less than 35 nm. The AgNPs exhibited a remarkable antibacterial activity against K. pneumoniae, E. coli, E. cloacae, S. aureus, S. epidermidis, and Shigella sp., and the MIC values ranged from 25 to 100 µg/mL. However, an exception was P. aeruginosa in which its MIC was >125 µg/mL. Conclusion: The results suggest that, the biosynthesized AgNPs by A. flavus could be utilized as a source of potent antibacterial agents in medicine and biotechnological applications.

Immunomodulatory effect of Moringa peregrina leaves, ex vivo and in vivo study.

This study was conducted to assess the in vivo and ex vivo immunomodulatory effect of the ethanol leaves extract of Moringa peregrina in Balb/c mice. For this study, five groups of 5 Balb/c mice were given a single acute subtoxic oral dose of the ethanolic extract at 1.13, 11.30, 23.40 and 113.4 mg/kg and the immunomodulatory effect was assessed on the 6th day following the ingestion. In the (non-functional) assessment, the effect of the extract on the body weight, relative lymphoid organ weight, splenic cellularity and peripheral blood hematologic parameters were evaluated. While in the immunomodulation assessment (functional), we investigated the effect of the extract on the proliferative capacity of splenic lymphocytes and peripheral T and B lymphocytes using mitogen blastogenesis, mixed allogeneic MLR and IgM-Plaque forming cells assays. The ingestion of M. peregrina extract caused a significant increase in the body weight, weight and number of cells of spleen and lymph nodes of the treated mice. Furthermore, the count of RBCs, WBCs, platelets, hemoglobin concentration and PCV % were increased by the extract treatment in a dose-dependent manner. M. peregrina enhanced the proliferative responses of splenic lymphocytes for both T cell and B-cell mitogens. Likewise, the mixed lymphocyte reaction MLR assay has revealed a T-cell dependent proliferation enhancement in the extract treated mice. Moreover, the oral administration of M. peregrina leaves extracts significantly increased PFCs/106 splenocytes in a dose-dependent manner. In conclusion, subtoxic acute doses of M. peregrina extract demonstrated significant potential as an immunomodulatory agent even at the lowest dose of 1.13 mg/kg.

The phytochemical content and antimicrobial activities of Malaysian Calophyllum canum (stem bark).

Recently there was huge increase in using of 'herbal products'. These can be defined as plants, parts of plants or extracts from plants that are used for curing disease. However, Calophyllum species is a tropical plant and it has been used in traditional medicine, the limitation in safety and effectiveness information could lead to serious health problems. Providing information for communities by evaluating the phytochemical contents, antioxidant, antimicrobial and cytotoxic activities will improve the therapeutic values. Three main Calophyllum canum fractions (none - high polar) were tested to find out the phenolic, flavonoid, flavonol content, DPPH radical scavenging, reducing power and chelating iron ions. Also were tested against Bacillus cereus, Staphylococcus aureus, Escherichia coli, Psedomonas aeruginosa, Candida albicans, and Cryptococcus neoformans. In addition, cytotoxic activity was assayed against lung cancer A549 cell line. The methanol fraction showed no bioactivity but achieved the highest amount of phenolic, flavonol and flavonoid contents, also it showed a significant result as antioxidant, reducing power and chelating agent. The n-hexane fraction achieved the minimum inhibitory concentration (MIC) value 12.5 µg. mL(-1) against B. cereus while the MIC value for DCM fraction was 25 µg. mL(-1). The DCM fraction was more active against S. aureus where the result was 50 µg. mL(-1) while the n-hexane fraction was 100 µg. mL(-1). The three main fractions have shown no activity against gram negative bacterial and fungal. The n-hexane and DCM fractions have shown cytotoxicity against lung cancer cell line; the 50% inhibition concentration (IC(50)) was 22 ± 2.64 and 32 ± 3.78 µg. mL(-1) respectively. The results were statistically significant (P < 0.05). Among the results, C. canum fractions proved to be effective against gram positive bacterial and anti-proliferation activity. Also it showed antioxidant activity as well. The results provided beneficial information for communities as well as can help to search for alternative drugs, and will contribute to establish safe and effective use of phytomedicines in the treatment of diseases.

Antimicrobial activity and essential oils of Curcuma aeruginosa, Curcuma mangga, and Zingiber cassumunar from Malaysia.

OBJECTIVE: To analyze the chemical composition of the essential oils of Curcuma aeruginosa (C. aeruginosa), Curcuma mangga (C. mangga), and Zingiber cassumunar (Z. cassumunar), and study their antimicrobial activity. METHODS: Essential oils obtained by steam distillation were analyzed by gas chromatography-mass spectrometry (GC-MS). The antimicrobial activity of the essential oils was evaluated against four bacteria: Bacillus cereus (B. cereus), Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa); and two fungi: Candida albicans (C. albicans) and Cyptococcus neoformans (C. neoformans), using disc-diffusion and broth microdilution methods. RESULTS: Cycloisolongifolene, 8,9-dehydro formyl (35.29%) and dihydrocostunolide (22.51%) were the major compounds in C. aeruginosa oil; whereas caryophyllene oxide (18.71%) and caryophyllene (12.69%) were the major compounds in C. mangga oil; and 2,6,9,9-tetramethyl-2,6,10-cycloundecatrien-1-one (60.77%) and α-caryophyllene (23.92%) were abundant in Z. cassumunar oil. The essential oils displayed varying degrees of antimicrobial activity against all tested microorganisms. C. mangga oil had the highest and most broad-spectrum activity by inhibiting all microorganisms tested, with C. neoformans being the most sensitive microorganism by having the lowest minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of 0.1 µL/mL. C. aeruginosa oil showed mild antimicrobial activity, whereas Z. cassumunar had very low or weak activity against the tested microorganisms. CONCLUSIONS: The preliminary results suggest promising antimicrobial properties of C. mangga and C. aeruginosa, which may be useful for food preservation, pharmaceutical treatment and natural therapies.

In vitro antimicrobial activity of mangrove plant Sonneratia alba.

OBJECTIVE: To investigate the antimicrobial property of mangrove plant Sonneratia alba (S. alba). METHODS: The antimicrobial activity was evaluated using disc diffusion and microdilution methods against six microorganisms. Soxhlet apparatus was used for extraction with a series of solvents, n-hexane, ethyl acetate and methanol in sequence of increasing polarity. RESULTS: Methanol extract appeared to be the most effective extract while n-hexane extract showed no activity. The antimicrobial activities were observed against the gram positive bacteria Staphylococcus aureus (S. aureus) and Bacillus cereus (B. cereus), the gram negative Escherichia coli (E. coli) and the yeast Cryptococcus neoformans. Pseudomonas aeruginosa and Candida albicans appeared to be not sensitive to the concentrations tested since no inhibition zone was observed. E. coli (17.5 mm) appeared to be the most sensitive strain followed by S. aureus (12.5 mm) and B. cereus (12.5 mm). CONCLUSIONS: From this study, it can be concluded that S. alba exhibits antimicrobial activities against certain microorganisms.

Antimicrobial activity of mangrove plant (Lumnitzera littorea).

OBJECTIVE: To investigate the antimicrobial activities of n-hexane, ethyl acetate and methanol extracts of the leaves of Lumnitzera littorea (L. littorea) against six human pathogenic microbes. METHODS: The antimicrobial activity was evaluated using disc diffusion and microdilution methods. RESULTS: The antimicrobial activities of the crude extracts were increased with increasing the concentration. It is clear that n-hexane extract was the most effective extract. Additionally, Gram positive Bacillus cereus (B. cereus) appear to be the most sensitive strain while Pseudomonas aeruginosa (P. aeruginosa) and the yeast strains (Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans)) appear to be resistance to the tested concentrations since no inhibition zone was observed. The inhibition of microbial growth at concentration as low as 0.04 mg/mL indicated the potent antimicrobial activity of L. littorea extracts. CONCLUSIONS: The obtained results are considered sufficient for further study to isolate the compounds responsible for the activity and suggesting the possibility of finding potent antibacterial agents from L. littorea extracts.

Antibacterial activity in vitro of Thymus capitatus from Jordan.

This study was carried out to evaluate the antibacterial activity of aqueous and organic extracts of Thymus capitatus L. (Lamiaceae) leaves and stems. Dried ground powder leaves and stems were extracted with water (aqueous extracts), ethanol, dichloromethane and hexane (Soxhlet extracts). The antibacterial activity of these extracts was evaluated against bacteria using disc diffusion method. The result obtained showed that the leaves had stronger antibacterial activity than the stems extracts. The ethanolic extract had the highest yield products and the high antibacterial activity than all other solvents. The results suggest that essential oil as non-polar organic compounds could be the main active compounds in this plant. Therefore the antibacterial activity of leaves ethanol extracts (LEE) was compared with essential oils leaves extracts (LEO) of T. capitatus. The LEO showed greater antibacterial activity than LEE. The LEO showed a broad spectrum of antibacterial activity and the Pseudomonas aeruginosa was the most sensitive bacteria.