Synergistic bactericidal activity of <Emphasis Type="Italic">Eremanthus erythropappus</Emphasis> oil or β-bisabolene with ampicillin against <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>

The activity of Eremanthus erythropappus oil (EO) and some of its compounds and their potential synergistic interaction with ampicillin against different strains of Staphylococcus aureus were investigated. Determination of chemical composition of EO by gas chromatography-mass spectrometry (GC/MS) and bioguided chemical fractionation led to the identification of β-bisabolene as the main active compound. A synergistic bactericidal activity of EO or β-bisabolene with ampicillin against Staphylococcus aureus was observed in a time-kill assay. EO and β- bisabolene have the potential to restore the effectiveness of ampicillin against resistant S. aureus.     

Phytochemical and Biological Profile of Essential Oils of Elionurus muticus (Spreng.) Growing in Northeastern Argentina

The purpose of this study was to investigate essential oils (EOs) from leaves of Elionurus muticus growing in Northeastern Argentina regarding their physicochemical profiles as well as their biological potential. Roots of a selected E. muticus population were investigated too. For this purpose, EOs of fresh materials were obtained by steam distillation and the chemical composition was characterized by gas chromatography GC/MS-FID. Antibacterial, antioxidant and eco-toxicity activities of the essential oils (EOs) were tested by in vitro assays. The EOs showed three E. muticus chemotypes: citral (neral+geranial), acorenone+bisabolone, acorenone+geranial. EO of roots of citral population contains mainly acorenone derivatives. EOs have high antibacterial effect against Staphylococcus aureus, being found minor antibacterial effect against Gram-negative bacteria. The half-maximal inhibitory concentration of EOs against DPPH⋅ were 7.1–30.0 mg/mL and the eco-toxicity was high with LD₅₀ <39 μg/mL. Based on the findings, given the high variability in their chemical composition and biological activity of E. muticus EO and the promising yields, it could be potentially chosen for industrial applications.     

Chemical Composition and Biological Screening of the Essential Oils of Micromeria macrosiphon and M. arganietorum (Lamiaceae)

The chemical composition and in vitro biological activities of the essential oil (EO) of Micromeria macrosiphon Coss. and M. arganietorum (J. Emb.) R. Morales, two Lamiaceae endemic to south Morocco, were investigated. GC/MS analysis resulted in the identification of 36 metabolites from the EO of M. macrosiphon, 45 from M. arganietorum. Borneol was the major metabolite in both oils and together with related derivatives such as camphor, accounted for 2/3 of the EO of M. macrosiphon, 1/3 of those of M. arganietorum. Pinene and terpinene derivatives were also present in high proportions. From a chemotaxonomic point of view, the composition of the examined samples may be related to those of other species endemic to Macaronesia. Both EOs showed significant toxicity towards liver HepG2 and melanoma B16 4A5 tumor cell lines at 100 μg/mL; however, they were also cytotoxic towards S17 normal cell lines, with a selectivity index <1. No antibacterial activity was noticed against 52 strains at 100 μg/mL.     

Anti-biofilm activities of essential oils rich in carvacrol and thymol against Salmonella Enteritidis

The aim of the present study was to determine the bioactive compounds in four essential oils (EO’s) from Origanum heracleoticum, Origanum vulgare, Thymus vulgaris and Thymus serpyllum and to assess their antimicrobial and anti-biofilm activity against Salmonella Enteritidis. Strains were previously characterized depending on the expression of the extracellular matrix components cellulose and curli fimbriae as rdar (red, dry and rough) and bdar morphotype (brown, dry and rough). This study revealed that the EO’s and EOC’s (carvacrol and thymol) investigated showed inhibition of biofilm formation at sub-minimum inhibitory concentration. Comparing the efficacy of EO’s and EOC’s in the inhibition of biofilm formation between the strains with different morphotype (rdar and bdar) did not show a statistically significant difference. Results related to the effectiveness of EO’s and EOC’s (the essential oil components, carvacrol and thymol) on eradication of preformed 48?h old biofilms indicated that biofilm reduction occurred in a dose-dependent manner over time.     

Chemical Composition,Antibacterial Screening and Cytotoxic Activity of Chiliadenus antiatlanticus (Asteraceae) Essential Oil

The chemical composition and in vitro antibacterial and cytotoxic activities of the essential oil (EO) of Chiliadenus antiatlanticus (Emb. & Maire) Gómiz, an asteraceous species endemic to the southwest of Morocco, were investigated. The EO yield was 1.07±0.28 %, twenty-seven metabolites were identified representing more than 96.4 % of the total composition. Camphor (35.7 %) and derivatives, borneol (4.9 %) and camphene (4.2 %) together with intermedeol (19.9 %), α-pinene (15.5 %) and (E)-pinocarveol (4.1 %) were the major constituents. An antibacterial activity was noticed against 24 strains (all Gram-positive) out of 71 at MICs values=100 μg/mL. The EO also showed significant toxicity towards liver HepG2 (55.8 % of cell viability) and melanoma B16 4A5 (41.6 % of cell viability) tumor cell lines at 100 μg/mL.     

Thyme and suico essential oils: promising natural tools for potato common scab control

• Potato common scab is a worldwide disease mainly caused by Streptomyces scabiei. It seriously affects potato crops by decreasing tuber quality. Essential oils (EO) are natural products with recognised antimicrobial properties. In this research, the antibacterial activities of thyme, oregano, suico and mint EO against S. scabiei were analysed.
• Infected tubers and soil samples were used for bacterial isolation; the obtained isolates were genetically identified. The chemical composition of the EO was determined by GC‐MS. The broth microdilution method was used to analyse antibacterial properties of EO.
• Thirty‐one bacterial isolates were obtained. The isolate chosen for antibacterial assays was morpho‐physiologically and genetically identified as S. scabiei. Thyme EO was mainly composed of thymol and o‐cymene; suico EO of dihydrotagetone, trans‐tagetone and verbenone; oregano EO of trans‐sabinene hydrate, thymol and ?‐terpinene; and mint EO of menthone and menthol. All the EO tested were effective against S. scabiei, but thyme and suico EO were the most successful, with a minimum inhibitory concentration of 0.068 g·l^?1 and 0.147 g·l^?1, respectively, and a minimum bactericidal concentration of 0.137 g·l^?1 and 0.147 g·l^?1, respectively. Scanning electron microscopy showed similar damage caused by both thyme and suico EO to the bacterial envelope. Total phenolic content of EO was not related to their antibacterial activity.
• Thyme and suico EO are effective antibacterial agents against S. scabiei, impeding bacterial viability and disturbing the bacterial cell envelope. These EO are promising tools for control of potato common scab.

     

Pseudotsuga menziesii (Pinaceae): Volatile Profiles,Antimicrobial Activity and Toxicological Evaluation of Its Essential Oil

The present article investigates the chemical composition of volatiles of essential oil (EO) and headspace (HS) fraction, as well as biological activities of EO obtained from needles with twigs of Pseudotsuga menziesii var. menziesii cultivated in Serbia. The major class of compounds was monoterpene hydrocarbons with α-terpinolene, sabinene and β-pinene (EO), and sabinene, α-terpinolene and β-pinene (HS) as the dominant volatiles. Tested EO exhibited mostly low antimicrobial potential against investigated strains (ATCC and respiratory isolates), where MICs ranged 1.25–20.00 mg/mL. Nevertheless, based on presented results, where antimicrobial testing was done for the first time on human respiratory system isolates, there is a potential of this EO to be used as an adjuvant in the treatment of human respiratory infections, especially those caused by Pseudomonas aeruginosa or Candida albicans strains. Regarding toxicological evaluation, EO showed moderate toxicity in Artemia salina toxicity bioassay (LC₅₀=347.41, after 24 h) as well as week toxicity against Drosophila melanogaster with the ability only to moderately delay larval and pupal development.     

Synergistic effect on anti-methicillin-resistant Staphylococcus aureus among combinations of α-mangostin-rich extract,lawsone methyl ether and ampicillin

α-Mangostin-rich extract (AME) exhibited satisfactory inhibitory activities against all tested MRSA strains, with minimum inhibitory concentrations (MICs) of 7·8–31·25 µg ml⁻¹, whereas lawsone methyl ether (LME) and ampicillin revealed weak antibacterial activity with MICs of 62·5–125 µg ml⁻¹. However, the combination of AME and LME showed synergistic effects against all tested MRSA strains with fractional inhibitory concentration index (FICI) values of 0·008–0·009, while the combination of AME and ampicillin, as well as LME and ampicillin produced synergistic effects with FICIs of 0·016–0·257. A time-kill assay against MRSA (DMST 20654 strain) revealed a 6-log reduction in CFU per ml, which completely inhibited bacterial growth for the combinations of AME and LME, AME and ampicillin, and LME and ampicillin at a 8-h incubation, while those against MRSA (2468 strain) were at 10-h incubation. The combination of α-mangostin and LME as well as the combinations of each compound with ampicillin synergized the alteration of membrane permeability. In addition, α-mangostin, LME and ampicillin inhibited the biofilm formation of MRSA. These findings indicated that the combinations of AME and LME or each of them in combination with ampicillin had enhanced antibacterial activity against MRSA. Therefore, these compounds might be used as the antibacterial cocktails for treatment of MRSA.     

In Vitro Trypanocidal and Antibacterial Activities of Essential Oils from Four Species of the Family Annonaceae

The objective of this study was to evaluate the chemical composition, and the trypanocidal and antibacterial activities of the essential oils from four species of Annonaceae: Bocageopsis multiflora (Mart .) R.E.Fr ., Duguetia quitarensis Benth ., Fusaea longifolia (Aubl. ) Saff ., and Guatteria punctata (Aubl .) R.A.Howard . The chemical composition of the essential oils from the aerial parts yielded 23, 20, 21 and 23 constituents, respectively, which were identified by GC/MS. The trypanocidal activity was evaluated against the amastigote and trypomastigote forms of T. cruzi. The antibacterial activity was evaluated by the microdilution method against enterohemorrhagic Escherichia coli, Pseudomonas aeruginosa, Streptococcus mutans, Streptococcus pyogenes, and methicillin‐resistant Staphylococcus aureus. The results of trypanocidal activity showed that the essential oils of the four species were active at the tested concentrations, with G. punctata essential oil being the most active, with IC₅₀=0.029 μg/mL, and selectivity index (SI)=32, being 34 times more active than the reference drug benznidazole. All EOs showed strong antibacterial activity (minimum inhibitory concentrations of 4.68–37.5 μg/mL) against strains of S. mutans.     

Chemical Composition and Antioxidant,Antiparasitic, Cytotoxicity and Antimicrobial Potential of the Algerian Limonium Oleifolium Mill. Essential Oil and Organic Extracts

This work aimed to investigate, for the first time, the chemical composition, antioxidant, antiparasitic, cytotoxicity, and antimicrobial activities of the aromatic plant Limonium oleifolium Mill. essential oil (EO) and organic extracts. L. oleifolium aerial parts essential oil was analyzed by GC-FID and GC-MS, and 46 constituents representing 98.25±1.12 % of the oil were identified. γ-Muurolene (10.81±0.07 %), cis-caryophyllene (7.71±0.06 %), o-cymene (7.07±0.01 %) and α-copaene (5.02±0.05 %) were the essential oil main compounds. The antioxidant activity of L. oleifolium EO and organic extracts (MeOH, CHCl₃, AcOEt, BuOH) was explored using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ABTS, β-carotene/linoleic acid, cupric reducing antioxidant capacity (CUPRAC), and ferric reducing power assays. The results showed that L. oleifolium EO exhibit antioxidant capacity (IC₅₀=17.40±1.32 μg/mL for DPPH assay, IC₅₀=29.82±1.08 μg/mL for β-carotene assay, IC₅₀=25.23±1.01 μg/mL for ABTS assay, IC₅₀=9.11±0.08 μg/mL for CUPRAC assay and IC₅₀=19.41±2.06 mg/mL for reducing power assay). Additionally, the EO showed significant activity against trophozoite form of Acanthamoeba castellanii (IC₅₀=7.48±0.41 μg/mL) and promastigote form of Leishmania amazonensis (IC₅₀=19.36±1.06 μg/mL) and low cytotoxicity on murine macrophages (LC₅₀ 90.23±1.09 μg/mL), as well as good antimicrobial activity against Staphylococcus aureus, Escherichia coli, Klebsiella oxytoca, and Pseudomonas aeruginosa. These results suggest that L. oleifolium essential oil is a valuable source of bioactive compounds presenting antioxidant, antiparasitic, and antimicrobial activities. Furthermore, it is considered nontoxic.     

Phytochemical, antimicrobial, antioxidant and antigenotoxic potentials of Cyperus rotundus extracts

The aqueous, ethyl acetate, methanolic and Total Oligomer Flavonoids (TOF) enriched extracts, obtained from the aerial parts of Cyperus rotundus, were investigated for their contents in phenolic compounds. Antioxidative activity using the NBT/riboflavin assay system, antimicrobial activity against Gram positive and Gram negative bacterial reference strains as well as antigenotoxic activity tested with the SOS chromotest assay were also studied. Significant antibacterial activity against reference strains; Staphylococcus aureus, Enterococcus faecalis, Salmonella enteritidis and Salmonella typhimurium, was detected in the presence of ethyl acetate and TOF enriched extracts. In addition to their antimicrobial activity, the same extracts showed a significant ability to inhibit nitroblue tetrazolium reduction by the superoxide radical in a non enzymatic O₂^.− generating system, and were also able to reduce significantly the genotoxicity induced by nifuroxazide and Aflatoxin B1. The antioxidant, antimicrobial and antigenotoxic activities exhibited by C. rotundus depend on the chemical composition of the tested extracts.     

Chemical profile and biological activities of Cedrelopsis grevei H. Baillon bark essential oil

Cedrelopsis grevei H. Baillon bark, endemic plant from Madagascar, is used in folk medicine for the treatment of rheumatism, muscular pain, and for its antifungal and antibiotic activities. In this paper, the phytochemical composition, antioxidant, antimicrobial, and cytotoxic activities of C. grevei bark essential oil (EO), its non-polar (I and II) and polar (III) fractions and its main compounds (ishwarane, β-elemene and α-copaene) were investigated.

The GC–MS analysis pointed out the presence of 36 components, representing about 80% as semi-quantitative characterization of the total. The presence of ishwarane, β-elemene and α-copaene as the main constituents highlighted its peculiar composition as a sesquiterpene-rich phytocomplex. Moreover, the quantification was performed for the first time by means of the experimental and predicted response factors (ERFs and PRFs, respectively).

As regards the biological activity, C. grevei EO and its fractions showed weak antioxidant activity against Trolox. The whole EO demonstrated instead considerable antimicrobial activity against Staphylococcus aureus and Staphylococcus epidermidis, while its polar-fraction evidenced an interesting bioactivity against Pseudomonas aeruginosa and Candida albicans. Finally, C. grevei EO and its fractions exhibited an interesting cytotoxic activity on human lung cancer cells (A549) and human colorectal cancer cells (CaCo-2).     

Chemical composition and antibacterial potential of Artemisia arborescens L. essential oil

This study was undertaken to characterize the essential oil (EO) of Artemisia arborescens growing wild in Sicily. EO, extracted by steam distillation, was examined for its chemical composition and for its capability to inhibit some food-borne pathogen bacteria. A total of 43 compounds (13 monoterpene hydrocarbons, 14 oxygenated monoterpenes, 10 sesquiterpene hydrocarbons, three oxygenated sesquiterpenes and less amount of other three compounds), which account 93.73% of the total oil, were identified by gas chromatography and gas chromatography–mass spectrometry. Oxygenated monoterpenes (57.32%) constituted the main fraction, with β-thujone as the main compound (45.04%), followed by the sesquiterpene hydrocarbon chamazulene (22.71%). Undiluted EO showed a large inhibition spectrum against strains of Listeria monocytogenes (34 out of 44), whilst it was ineffective against enterobacteria and salmonellas. The minimum inhibition concentration (MIC) was evaluated for the two most sensitive strains (L. monocytogenes 186 and 7BO) at two cellular concentrations (10⁶ and 10⁷ CFU ml⁻¹). The lowest MIC (0.625 μl ml⁻¹, dilution of oil with acetone) was found for strain L. monocytogenes 186 at 10⁶ CFU ml⁻¹.     

Characterization of typical and atypical enteroaggregative Escherichia coli in Kagoshima,Japan: biofilm formation and acid resistance

EAEC is increasingly recognized as an emerging enteric pathogen. Typical EAEC expressing the AggR regulon have been proven to be an important cause of childhood diarrhea in industrialized countries as well as in the developing world, while atypical EAEC without this regulon have not been thoroughly investigated. To investigate the bacteriological characteristics of EAEC, including both typical and atypical strains in Kagoshima, Japan, 2417 E. coli strains from Japanese children with diarrhea were screened by a quantitative biofilm assay to detect possible EAEC strains, resulting in the identification of 102 (4.2%) of these strains by the HEp‐2 cell adherence test. Virulence gene patterns, PFGE analysis and O‐serogrouping demonstrated the heterogeneity of the EAEC. The EAEC strains were classified into two groups: typical EAEC with aggR (74.5%, 76/102) and atypical EAEC without aggR (25.5%, 26/102). There was no significant difference between the typical EAEC strains (median OD₅₇₀= 0.73) and the atypical strains (median OD₅₇₀= 0.61) in biofilm formation (P= 0.17). Incidences of resistance against ampicillin, cefotaxime and tetracycline were significantly higher in the typical EAEC strains than the atypical EAEC strains (84.2% vs. 53.8%, 36.8% vs. 7.7% and 93.4% vs. 73.1%, respectively, P < 0.05). The typical EAEC strains showed significantly higher resistance ratios against HCl and lactate than the atypical strains (94.7% vs. 61.5% and 92.1% vs. 57.7%, respectively, P < 0.001). To investigate the pathogenicity of not only typical but also atypical EAEC, further bacteriological and epidemiologic studies including atypical EAEC are needed.     

Phytochemical evaluation and in vitro antifungal activities of Melaleuca styphelioides leaves: comparison between volatile and non-volatile extracts

Abstract

Melaleuca styphelioides is considered as medicinal plant. This study was carried out to evaluate for the first time the phytochemical composition and to compare the antifungal activities of essential oil (EOs), methanol and aqueous extracts of M. styphelioides Sm. leaves against three fungi (Aspergillus niger, Rhizopus nigricans and Penicillium digitatum). A total of 10 components of the EO were identified, with the principal compound being methyl eugenol (87.2%). Results of the phytochemical analysis of leaves extract exhibited the presence of different phytoconstituents (phenolic compounds, flavonoids, tannins and anthocyanins). Volatile and non-volatile extracts were found to express dose-dependent inhibition against all tested fungi. Indeed, the EO oil showed significant inhibition of fungal growth and the IC₅₀ was 2.08?µL/mL for A. niger indicating that M. styphelioides leaf EO was particularly effective against this pathogen. The most susceptible species for the aqueous extract was P. digitatum (IC₅₀= 9.54?mg/mL) whereas R. nigricans was found to be more susceptible to the methanolic extract (IC₅₀= 8.31?mg/mL). Thus, the EO and aqueous as well as methanol extracts of M. styphelioides leaves possess antifungal activity and hence, it can be suggested for use in the food or pharmaceutical industries as an alternative to chemical preservatives.     

Artemisia arborescens Essential Oil Composition,Enantiomeric Distribution,and Antimicrobial Activity from Different Wild Populations from the Mediterranean Area

Aerial parts of Artemisia arborescens were collected from different sites of the Mediterranean area (southwestern Algeria and southern Italy) and the chemical composition of their essential oil (EO) extracted by hydrodistillation was studied by both gas chromatography (GC) equipped with an enantioselective capillary column and GC/mass spectrometry (GC/MS). The EOs obtained were tested against several Listeria monocytogenes strains. Using GC and GC/MS, 41 compounds were identified, accounting for 96.0 – 98.8% of the total EO. All EOs showed a similar terpene profile, which was rich in chamazulene, β‐thujone, and camphor. However, the concentration of such compounds varied among the EOs. A. arborescens EO inhibited up to 83.3% of the L. monocytogenes strains, but the inhibitory spectrum varied among the EOs, with those from Algeria showing a higher inhibition degree than the Italian EOs. Such effect likely depended on the ketone (β‐thujone + camphor) content of the EO. The differences in the EO composition support the hypothesis that A. arborescens has at least two different chemotypes: a β‐thujone and a chamazulene type. The EO inhibitory spectrum indicates the A. arborescens EO as a valuable option in the control of the food‐borne pathogens.     

Thymbra capitata essential oil has a significant antimicrobial activity against methicillin-resistant Staphylococcus aureus pre-formed biofilms

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant opportunistic pathogen with a great ability to form biofilms. Herein, the antimicrobial potential of Thymbra capitata essential oil (EO) against MRSA biofilms was investigated. The determination of the minimum inhibitory concentration (MIC) and the minimum lethal concentration (MLC) of the T. capitata EO was first investigated on a group of clinical isolates from septicaemias, diabetic foot ulcers and osteomyelitis. Biofilms were incubated with the EO at the MLC and its anti-biofilm potential was investigated. A strong antimicrobial activity was observed, with MIC and MLC values between 0·32 and 0·64 mg l⁻¹. However, the concentration of EO necessary for the eradication of planktonic cells was insufficient to significantly reduce the biofilm biomass of some isolates. Nevertheless, cell culturability and overall cellular metabolism was strongly reduced in all biofilms tested, only when the EO was tested. Contrary to the tested antibiotics, T. capitata EO showed a significant antimicrobial activity against MRSA biofilms, by reducing cellular metabolism and cellular culturability.     

Anti-adhesion activity of two biosurfactants produced by <Emphasis Type="Italic">Bacillus</Emphasis> spp. prevents biofilm formation of human bacterial pathogens

In this work, two biosurfactant-producing strains, Bacillus subtilis and Bacillus licheniformis, have been characterized. Both strains were able to grow at high salinity conditions and produce biosurfactants up to 10% NaCl. Both extracted-enriched biosurfactants showed good surface tension reduction of water, from 72 to 26–30 mN/m, low critical micelle concentration, and high resistance to pH and salinity. The potential of the two lipopeptide biosurfactants at inhibiting biofilm adhesion of pathogenic bacteria was demonstrated by using the MBEC device. The two biosurfactants showed interesting specific anti-adhesion activity being able to inhibit selectively biofilm formation of two pathogenic strains. In particular, Escherichia coli CFT073 and Staphylococcus aureus ATCC 29213 biofilm formation was decreased of 97% and 90%, respectively. The V9T14 biosurfactant active on the Gram-negative strain was ineffective against the Gram-positive and the opposite for the V19T21. This activity was observed either by coating the polystyrene surface or by adding the biosurfactant to the inoculum. Two fractions from each purified biosurfactant, obtained by flash chromatography, fractions (I) and (II), showed that fraction (II), belonging to fengycin-like family, was responsible for the anti-adhesion activity against biofilm of both strains.     

Casearia sylvestris Essential Oil Degradation Products Generated by Leaf Processing

Casearia sylvestris is an endemic tree of the Latin America that the essential oil (EO) has anti-inflammatory and gastroprotective actions. This study evaluates the chemical composition of the EO from the volatile fractions of in natura, fresh, and dried C. sylvestris var. sylvestris and var. lingua leaves. For both varieties, the dried leaves presented higher EO yield as compared to fresh leaves. The major EO chemical components were (E)-caryophyllene, α-humulene, germacrene D, bicyclogermacrene, spathulenol, caryophyllene oxide, and humulene epoxide II. In both varieties, the content of sesquiterpene hydrocarbons decreased and oxygenated sesquiterpenes increased on going from in natura to fresh and dried leaves, which indicated that leaf drying and hydrodistillation modified the volatile composition. The results also suggested that bicyclogermacrene and (E)-caryophyllene were oxidized during processing, to generate spathulenol and caryophyllene oxide, respectively. C. sylvestris varieties and in natura, fresh, and dried leaves differed in terms of the chemical composition of volatiles, which could affect the EO biological activities.