Geovany Amorim Gomes

Effects of Myrcia ovata Cambess. essential oil on planktonic growth of gastrointestinal microorganisms and biofilm formation of Enterococcus faecalis

The essential oil from the leaves of Myrcia ovata Cambess., commonly used in Brazil for the treatment of gastric illnesses, was screened for antimicrobial activity and action in the formation of microbial biofilms by Enterococcus faecalis. The oil was obtained by hydrodistillation using a clevenger-type system. Its chemical composition was analyzed using GC and GC-MS. Both MIC and MBC of the essential oil were determined by broth microdilution techniques and agar dilution method. The essential oil showed antimicrobial activity against E. faecalis, Escherichia coli, Pseudomonas aeruginosa, Salmonella choleraesuis, Staphylococcus aureus, Streptococcus pneumoniae and Candida parapsilosis. The results showed that the essential oil of M. ovata Cambess. was effective against the formation of biofilm by E. faecalis when compared with the control. Four volatile compounds, representing 92.1 % of the oil, were identified and geranial was the major component (50.4 %). At the best of our knowledge, this is the first report of the chemical composition and antimicrobial activity of the essential oil from leaves of M. ovata.

Evaluation of the combined effect of thymol, carvacrol and (E)-cinnamaldehyde on Amblyomma sculptum (Acari: Ixodidae) and Dermacentor nitens (Acari: Ixodidae) larvae

This study aimed at assessing the combined effect of thymol, carvacrol and (E)-cinnamaldehyde on Amblyomma sculptum and Dermacentor nitens larvae. The effects resulting from treatments were evaluated by means of the modified larval packet test. In order to determine the LC50, components of essential oils, the monoterpenes thymol, carvacrol and phenylpropanoid (E)-cinnamaldehyde were individually tested at different concentrations. After determining the LC50, each essential oil component was separately evaluated and then combined with another substance at a 1:1 proportion at the LC50 concentration and at 1/2 and 1/4 of the LC50. For A. sculptum, the lowest LC50 value was obtained for (E)-cinnamaldehyde (1.40 mg/ml), followed by thymol (2.04 mg/ml) and carvacrol (3.49 mg/ml). The same order of effectiveness was observed for D. nitens, with values of 1.68, 2.17 and 3.33 mg/ml, respectively. In the evaluation of component associations of essential oils against A. sculptum larvae, only the combinations between carvacrol and thymol (LC50) and carvacrol and (E)-cinnamaldehyde (1/4 LC50) presented a moderate synergetic effect. In turn, for D. nitens larvae, the combinations between thymol and carvacrol (LC50 and 1/2 LC50) presented a synergetic effect, while the others presented an additive or antagonistic effect. Therefore, it can be concluded that the combination of thymol and carvacrol (LC50) has a moderate synergetic effect against A. sculptum larvae, while thymol, combined with carvacrol (LC50 and 1/2 LC50), has a synergetic effect against D. nitens larvae.

Acaricidal activity of essential oil from Lippia sidoides on unengorged larvae and nymphs of Rhipicephalus sanguineus (Acari: Ixodidae) and Amblyomma cajennense (Acari: Ixodidae).

The aims of this work were to identify the compounds and to investigate the acaricidal activity of the essential oil of Lippia sidoides for unengorged larvae and nymphs of Rhipicephalus sanguineus and Amblyomma cajennense. The oil was analyzed by gas chromatography and gas chromatography/mass spectrometry. In total, 22 compounds comprising 98.5% of the total peak area were identified. The major constituent of the essential oil was thymol (69.9%). The acaricidal activity against larvae and nymphs was assessed using a modified larval packet test. In all experiments, oils were tested at concentrations of 2.35, 4.70, 9.40 14.10 and 18.80 mg/mL. The mortalities of larvae and nymphs of R. sanguineus were 20.6, 47.8, 73.6, 99.5 and 99.0% and 12.0, 50.0, 76.3, 96.0 and 96.1%, respectively. For larvae and nymphs of A. cajennense the rates of mortality were 41.9, 63.3, 77.8, 82.5 and 100.0% and 0.0, 32.8, 64.8, 71.1 and 94.0%, respectively. The LC 90 values of the L. sidoides oil were 11.56 and 12.97 mg/mL for larvae and nymphs of R. sanguineus and 15.70 and 18.52 mg/mL for larvae and nymphs of A. cajennense, respectively. The essential oil from L. sidoides has acaricidal activity on unengorged larvae and nymphs of R. sanguineus and A. cajennense.

Constituintes químicos e atividade antioxidante de folhas e galhos de Eugenia copacabanensis Kiaersk (Myrtaceae)

Phytochemical investigation of Eugenia copacabanensis allowed for the isolation and identification of following compounds: β-sitosterol, β-sitosterol-glucoside, eight triterpenes, (mixture of α- and β-amyrins, ursolic acid, 30-hydroxy-ursolic acid, betulin, friedelin, friedelan-3,4-lactone, and taraxerol), a mixture of three sesquiterpenes, (clovandiol, globulol, and viridiflorol), three flavonoids (kaempferol-3-O-β-D-rhamnoside, quercetin-3-O-α-L-arabinoside, and quercetin), and a mixture of four coumaroyl esters (octacosanyl, heptacosanyl, hexacosanyl, and tetracosanyl coumarates). The structures of these compounds were assigned based on comparison with literature data and spectroscopic analysis, including analysis by two-dimensional NMR techniques. Total phenolic content and total flavonoids were evaluated. Antioxidant activities of methanol extracts and fractions were measured by the 1,2-diphenyl-2-picryl-hidrazyl free radical scavenging assay.

In Vitro Antibacterial and Antibiofilm Activity of Lippia alba Essential Oil, Citral, and Carvone against Staphylococcus aureus

In vitro antimicrobial and antibiofilm activities of the Lippia alba essential oil and its major components (citral and carvone) against Staphylococcus aureus were investigated. Essential oils (LA1EO, LA2EO, and LA3EO) were extracted from the aerial parts of three L. alba specimens by hydrodistillation and analyzed by gas chromatography coupled to a mass spectrometer. Minimum Inhibitory Concentrations (MIC) and Minimum Bacterial Concentration (MBC) were determined by the microdilution method. For the antibiofilm assays, the biomass formation in the biofilm was evaluated by the microtiter-plate technique with the crystal violet (CV) assay and the viability of the bacterial cells was analyzed. All oils and their major components presented antibacterial activity, and the lowest MIC and MBC values were 0.5 mg mL−1 when LA1EO and citral were used. Potential inhibition (100%) of S. aureus biofilm formation at the concentration of 0.5 mg mL−1 of all EOs was observed. However, the elimination of biofilm cells was confirmed at concentrations of 1 mg mL−1, 2 mg mL−1, 2 mg mL−1, and 0.5 mg mL−1 for LA1EO, LA2EO, LA3EO, and citral, respectively. The results obtained in the present research point to the promising antibacterial and antibiofilm potential of L. alba EOs against S. aureus, a species of recognized clinical interest.

Lethal and sublethal effects of essential oil of Lippia sidoides (Verbenaceae) and monoterpenes on Chagas’ disease vector Rhodnius prolixus

The aim of this study was to identify the composition of the essential oil from leaves of Lippia sidoides (EOLS), a typical shrub commonly found in the dry northeast of Brazil, popularly known as “alecrim-pimenta”. Additionally, we investigated the nymphicidal, ovicidal, phagoinhibitory and excretion effects of EOLS, its major constituent thymol and its isomer carvacrol, on fourth instar nymphs and eggs of Rhodnius prolixus, the Chagas’ disease vector. The nymphicidal and ovicidal activity of thymol, carvacrol, and EOLS was assessed by tests using impregnated Petri dishes. The lethal concentration values (LC50) for EOLS, carvacrol, and thymol were 54.48, 32.98, and 9.38 mg/cm2, respectively. The ovicidal test showed that both carvacrol and thymol (50 mg/cm2) inhibited hatching (50% and 23.3%, respectively), while treatments with 10 mg/cm2 or 50 mg/cm2 EOLS did not affect the hatching rate at all (80% and 90%, respectively). We observed an anti-feeding effect in insects fed with blood containing natural products at the higher concentrations (100 µg/mL). Finally, excretion rate was affected by EOLS and carvacrol, but not by thymol. These findings offer novel insights into basic physiological processes that make the tested natural compounds interesting candidates for new types of insecticides.

Antibacterial and Antibiofilm Activities of Cinnamomum Sp. Essential Oil and Cinnamaldehyde: Antimicrobial Activities

To assess the activities of essential oils derived from the trunk bark of Cinnamomum zeylanicum (EOCz) and Cinnamomum cassia (EOCc) as well as cinnamaldehyde on bacterial biofilms of clinical interest. Antimicrobial activity was assessed by the broth microdilution method to determine minimum inhibitory concentrations (MICs). Antibiofilm activity was assessed by quantifying the biomass and determining the number of viable cells. The chemical composition of the essential oils was determined. The results showed that the major component of EOCz and EOCc was cinnamaldehyde. For the assayed substances, biofilm biomasses were reduced by up to 99.9%, and Streptococcus pyogenes, Pseudomonas aeruginosa, and Escherichia coli biofilms were sensitive to all of the concentrations and substances analysed. In cell viability tests, 2 mg/ml of cinnamaldehyde reduced the number of viable cells by 5.74 Log CFU/ml. EOCz, EOCc, and cinnamaldehyde exhibited antimicrobial and antibiofilm activities. This work describes substances with potential use against infections caused by bacterial biofilms.

In vitro assessment of the acaricidal activity of carvacrol, thymol, eugenol and their acetylated derivatives on Rhipicephalus microplus (Acari: Ixodidae)

This study reports the comparative evaluation of the activity of carvacrol, thymol, eugenol and their respective acetylated derivatives (carvacrol acetate, thymol acetate and eugenol acetate) on Rhipicephalus microplus, to verify the possible influence of the acetate group. The acetylated derivatives were prepared from reactions of the phenolic compounds with acetic anhydride/pyridine. The formation of the products was confirmed by analysis of hydrogen and carbon-13 nuclear magnetic resonance (1H and 13C NMR) spectra. The larval packet test was used to evaluate the acaricidal activity, with concentrations of 0.312, 0.625, 1.25, 2.5, 5.0, 7.5, 10.0 and 15.0 mg/mL. Thymol and carvacrol resulted in 100% mortality starting at the concentration of 2.5 mg/mL, while the same was observed for carvacrol acetate starting at the concentration of 5.0 mg/mL. For the other treatments, 100% mortality was only achieved in the groups treated with the highest concentration (15.0 mg/mL). The LC50 and LC90 values (mg/mL) of carvacrol acetate (2.49, 4.21), thymol acetate (2.97, 8.52) and eugenol acetate (4.25, 13.10) were higher than those for the corresponding precursor molecules carvacrol (0.83, 2.02), thymol (1.26, 2.21) and eugenol (2.77, 5.35). The acetylation process did not enhance the activity of these substances on unengorged larvae of R. microplus, since the precursor substances (carvacrol, thymol and eugenol) had greater efficacy.