Staphylococcus aureus is a serious human pathogen that is highly adaptive to environmental conditions and rapidly develops antibiotic resistance. The use of efflux pumps to reduce antibiotic concentrations at the intracellular level is one of the main mechanisms by which bacteria develop antibiotic resistance. The management of efflux pumps, specifically NorA, which is expressed by S. aureus strains, is a valuable strategy for restoring susceptibility in strains resistant to antibacterial agents. In recent years, many studies have focused on searching for natural substances that can reverse efflux pump-mediated resistance in S. aureus. Extracts and compounds obtained from plants can be efficient efflux pump inhibitors (EPIs) and represent a potentially patient-friendly strategy for controlling S. aureus. In the present study, we evaluated the ability of essential oils, petroleum ether extracts, dichloromethane extract (DCME) and six compounds isolated from the heartwood of Pilgerodendron uviferum (Cupressaceae) and two synthetic derivatives to inhibit efflux in NorA pumps in the following three S. aureus strains: K2378, which overexpressed the norA gene (norA++), K1902 (norA-deleted, ΔnorA) and the parental strain, NCTC 8325-4. Efflux activity was evaluated using a fluorometric method that measured the accumulation of the universal efflux pump substrate ethidium bromide (EtBr). Only DCME and the compounds 15-copaenol and epi-cubenol inhibited EtBr efflux by K2378. Even the lowest concentration of 15-copaenol exhibited a stronger inhibitory effect than carbonyl cyanide m-chlorophenyl hydrazone on EtBr efflux by K2378. 15-copaenal only showed inhibition of EtBr efflux in K2378 cells at 125 μg/mL, but not superior to the control inhibitor and 15-copaenyl acetate exerted no intrinsic EPI activity against K2378. Fractional inhibitory concentration index (FICI) values obtained in the checkerboard assays, indicated that all combinations between DCME, epi-cubenol and 15-copaenol, and tested antibiotics showed a synergistic effect in wild type, norA⁺⁺ and ΔnorA strains. Moreover, those were not toxic for the HeLa cell line at concentrations in which the synergistic effect and inhibitory activity of efflux pumps was determined. Other extracts and compounds obtained from P. uviferum did not display EtBr efflux-inhibiting activity against the evaluated S. aureus strains.

Conventional asphalt mixtures used for road paving require high manufacturing temperatures and therefore high energy expenditure, which has a negative environmental impact and creates risk in the workplace owing to high emissions of pollutants, greenhouse gases, and toxic fumes. Reducing energy consumption and emissions is a continuous challenge for the asphalt industry. Previous studies have focused on the reduction of emissions without characterizing their composition, and detailed characterization of volatile organic compounds (VOCs) and semi-volatile organic compounds (SVOCs) in asphalt fumes is scarce. This communication describes the characterization and evaluation of VOCs and SVOCs from asphalt mixtures prepared at lower production temperatures using natural zeolite; in some cases, reclaimed asphalt pavement (RAP) was used. Fumes were extracted from different asphalt mix preparations using a gas syringe and then injected into hermetic gas sample bags. The compounds present in the fumes were sampled with a fiber and analyzed by gas-liquid chromatography coupled to mass spectrometry (GC/MS). In general, the preparation of warm mix asphalts (WMA) using RAP and natural zeolite as aggregates showed beneficial effects, reducing VOCs and SVOCs compared to hot mix asphalts (HMA). The fumes captured presented a similar composition to those from HMA, consisting principally of saturated and unsaturated aliphatic hydrocarbons and aromatic compounds but with few halogenated compounds and no polycyclic aromatic hydrocarbons. Thus, the paving mixtures described here are a friendlier alternative for the environment and for the health of road workers, in addition to permitting the re-use of RAP.

The beetle Hylastinus obscurus Marsham (Coleoptera: Curculionidae), endemic to Europe and Northern Africa, is one of the most important red clover pests in Chile. As commercial insecticides are less effective against this pest, plant secondary metabolites have been considered as an alternative for its control. Here, we have investigated the chemical composition of essential oil (EO), petroleum ether extract (PEE), and dichloromethane extract (DCME) from Pilgerodendron uviferum heartwood. Additionally, the effects of EO and extracts on the feeding behavior (% of weight shift) of H. obscurus have been evaluated. The composition of EO, PEE, and DCME were analyzed using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The results showed the presence of a similar mixture of sesquiterpenes in the essential oil and in both of the extracts, which accounted for circa 60% of the total mixture. Sesquiterpenes were further isolated using chromatographic methods and were structurally characterized by optical rotation, GC–MS, FTIR, and 1D and 2D NMR experiments. The physicochemical properties of the isolated sesquiterpenes, including lipophilicity and vapor pressure, were also determined. The sesquiterpenes were identified as the following: (−)-trans-calamenene (1), cadalene (2), (−)-cubenol (3), (−)-epi-cubenol (4), (−)-torreyol (5), and (−)-15-copaenol (6). The antifeedant activity of EO, extracts, and isolated sesquiterpenes were evaluated using artificial diets in a non-choice test. Relative to the control, the EO, DCME extract, and the isolated sesquiterpenes, namely, (−)-trans-calamenene (1), cadalene (2), and (5) torreyol, were found to be the most effective treatments against H. obscurus. Our study showed that the compounds occurring in P. uviferum heartwood were effective in reducing the adult growth of H. obscurus. The physicochemical properties of the isolated sesquiterpenes might have been associated with antifeedant effects.

Adverse effects caused by synthetic pesticides have increased interest in plant-derived insecticidal compounds, in particular essential oils, as a more compatible and ecofriendly alternative for pest control of economic importance. For this reason, the essential oil isolated from leaves and shoots of Drimys winteri (J.R. Forster & G. Forster)—also named canelo (CEO)—was investigated for its chemical profile and insecticidal action against Acanthoscelides obtectus (Say)—one of the most important post-harvest pests of dry beans in the world—and Aegorhinus superciliosus (Guérin)—a significant pest of fruit trees in Chile. The analysis by gas chromatography, paired with mass spectrometry (GC/MS) determined 56 compounds, corresponding to 92.28% of the detected compounds. Elemol (13.54%), γ-eudesmol (11.42%), β-eudesmol (8.49%), α-eudesmol (6.39%), α-pinene (7.92%) and β-pinene (5.17%) were the most abundant. Regarding the bioactivity of the CEO, the results demonstrated toxicological effects against A. obtectus. A concentration of 158.3 µL L⁻¹ had a mortality rate of 94% after 24 h exposure. The LC₅₀ and LC₉₀ values at 24 h were 60.1 and 163.0 µL L⁻¹. Moreover, behavioral bioassays showed a repellent effect against A. superciliosus with a dose of one microliter of CEO. Both sexes of the raspberry weevil stayed for very short times in the treated area with the oil (<0.8 min), showing a homogeneous repellency in the species. The overall data suggest that canelo leaves and shoots essential oil has an insecticide effect and is worth exploring to better understand the synergistic relationship between the compounds present in the essential oil.