Analysis of cell death in Bacillus subtilis caused by sesquiterpenes from Chrysopogon zizanioides (L.) Roberty.

Recently, the antibacterial effects of essential oils have been investigated in addition to their therapeutic purposes. Owing to their hydrophobic nature, they are thought to perturb the integrity of the bacterial cell membrane, leading to cell death. Against such antibiotic challenges, bacteria develop mechanisms for cell envelope stress responses (CESR). In Bacillus subtilis, a gram-positive sporulating soil bacterium, the extracytoplasmic function (ECF) sigma factor-mediated response system plays a pivotal role in CESR. Among them, σM is strongly involved in response to cell envelope stress, including a shortage of available bactoprenol. Vetiver essential oil, a product of Chrysopogon zizanioides (L.) Roberty root, is also known to possess bactericidal activity. σM was exclusively and strongly induced when the cells were exposed to Vetiver extract, and depletion of multi-ECF sigma factors (ΔsigM, ΔsigW, ΔsigX, and ΔsigV) enhanced sensitivity to it. From this quadruple mutant strain, the suppressor strains, which restored resistance to the bactericidal activity of Vetiver extract, emerged, although attempts to obtain resistant strains from the wild type did not succeed. Whole-genome resequencing of the suppressor strains and genetic analysis revealed inactivation of xseB or pnpA, which code for exodeoxyribonuclease or polynucleotide phosphorylase, respectively. This allowed the quadruple mutant strain to escape from cell death caused by Vetiver extract. Composition analysis suggested that the sesquiterpene, khusimol, might contribute to the bactericidal activity of the Vetiver extract.

Evaluation of Vetiver Volatile Compound Production under Aeroponic-Grown Conditions for the Perfume Industry.

Vetiver (Chrysopogon zizanioides (L.) Roberty) is a major tropical perfume crop. Access to its essential oil (EO)-filled roots is nevertheless cumbersome and land-damaging. This study, therefore, evaluated the potential of vetiver cultivation under soilless high-pressure aeroponics (HPA) for volatile organic compound (VOC) production. The VOC accumulation in the roots was investigated by transmission electron microscopy, and the composition of these VOCs was analyzed by gas chromatography coupled with mass spectrometry (GC/MS) after sampling by headspace solid-phase microextraction (HS-SPME). The HPA-grown plants were compared to plants that had been grown in potting soil and under axenic conditions. The HPA-grown plants were stunted, demonstrating less root biomass than the plants that had been grown in potting soil. The roots were slender, thinner, more tapered, and lacked the typical vetiver fragrance. HPA cultivation massively impaired the accumulation of the less-volatile hydrocarbon and oxygenated sesquiterpenes that normally form most of the VOCs. The axenic, tissue-cultured plants followed a similar and more exacerbated trend. Ultrastructural analyses revealed that the HPA conditions altered root ontogeny, whereby the roots contained fewer EO-accumulating cells and hosted fewer and more immature intracellular EO droplets. These preliminary results allowed to conclude that HPA-cultivated vetiver suffers from altered development and root ontology disorders that prevent EO accumulation.

Thermoplastic starch nanocomposites using cellulose-rich Chrysopogon zizanioides nanofibers.

Green thermoplastic starch (TPS) nanocomposite films aided by cellulose nanofibers (CNFs) from Chrysopogon zizanioides roots were developed and characterized. When compared to other lignocellulosic fibers, Chrysopogon zizanioides roots revealed exceptionally high cellulose content (~48%). CNFs were separated using an environmentally friendly acid isolation technique that included three stages: (i) alkali treatment; (ii) bleaching; and (iii) mild acid hydrolysis using oxalic acid in an autoclave. Following that, green nanocomposite films were made from potato starch using the solution casting process, by which we used glycerol (30 wt%) to make thermoplastic starch. Then, cellulose nanofibers in different concentrations (0, 1, 2, 3, 4 wt%) were added to the thermoplastic starch matrix. The isolated CNFs had diameters in the range of 17-27 nm. Besides, these nanostructures presented a very high crystallinity index (~65%), thereby enhanced the thermal stability. TPS/CNF green nanocomposites containing 3 wt% CNFs had exceptional tensile strength (~161%), tensile modulus (~167%), thermal stability, and crystallinity. As a result, nanocomposite films made of starch and cellulose nanofibers (3 wt%) extracted from Chrysopogon zizanioides roots would be alternatives for sustainable packaging. It can be concluded that Chrysopogon zizanioides roots have high potential for polymer industry.

Phytochemical Screening, Antioxidant Assay and Cytotoxic Profile for Different Extracts of Chrysopogon zizanioides Roots.

The Chrysopogon zizanioides plant possesses multiple traditional uses, especially in therapeutics, but only a few articles have reported its biological activity. Hence, the present study was planned to explore the phytochemical constituents, cytotoxic potential, radical scavenging activity, and GC/MS (Gas chromatography & Mass spectrometry) analysis of the vetiver root extracts. The roots extracted with different solvents exhibited more significant phytochemical constituents in polar solvents in comparison to non-polar ones, favoring the extraction of a greater number of components in highly polar solvents. All the extracts were tested for their cytotoxicity using SRB (Sulforhodamine B) assay. They confirmed ethanolic extract as a potent extract with GI50 56±0.5 µg/ml in oral cancer (SCC-29B) along with no cytotoxicity in healthy cells (Vero cells), making it a safer therapeutic option in comparison to standard Adriamycin. This extract was also analyzed for its antioxidant potential by DPPH (1,1-Diphenyl-2-picrylhydrazyl) assay with IC50 value 10.73 µg/ml, which was quite comparable to Ascorbic acid having IC50 value 4.61 µg/ml. The quantitative analysis of ethanolic extract exhibited 107 compounds amongst which Khusenic acid, Ascorbic acid, Junipen, gamma-Himachalene, alpha-Guaiene were the majorly occurring compounds that can be explored further for their cytotoxic activity.

Root essential oil of Chrysopogon zizanioides relaxes rat isolated thoracic aorta - an ex vivo approach.

Root of Chrysopogon zizanioides (L.) Roberty has been used in Siddha system of medicine to treat hypertension. The present study was therefore to investigate the vasorelaxation effect of root essential oil of C. zizanioides using rat isolated thoracic aortic rings. Chemical characterization of root essential oil was carried out using Gas Chromatography-Flame Ionization Detector (GC-FID) and Gas Chromatography-Mass Spectrometry (GC-MS). Essential oil nanoemulsion (EONE) was prepared and characterized. Vasorelaxant effect of EONE in endothelium-intact aortic rings precontracted with phenylephrine (PE) (1 µM) or KCl (80 mM) was investigated. Role of Ca2+, nitric oxide and K+ channels in precontracted aortic rings were investigated to elucidate the mechanism of action of the essential oil. Further, the role of muscarinic and prostacyclin receptors in EONE induced relaxation was studied. The EONE significantly induced relaxation (Emax 77.1 ± 4.87%) in PE precontracted aortic rings. The nitric oxide synthase, and cyclooxygenase inhibitors and potassium channel blockers have not significantly inhibited the vasorelaxation induced by EONE. However, EONE induced relaxation in precontracted endothelium-intact aortic rings was significantly inhibited by muscarinic receptor and calcium channel. The root essential oil of C. zizanioides possesses vasorelaxant effect through muscarinic pathway as well as acts as calcium channel blocker.

Metabolic response of vetiver grass (Chrysopogon zizanioides) to acid mine drainage.

Acid mine drainage (AMD) is a sulfuric discharge containing metals and particulates that can spread to nearby water sources, imposing toxicity and physical stress to living things. We have shown that vetiver grass (Chrysopogon zizanioides) is capable of tolerating and treating AMD-impacted water from the abandoned Tab-Simco mining site from southern Illinois, though little is known about its tolerance mechanisms. We conducted metabolomic analyses of vetiver shoots and roots after relatively short- and long-term periods of exposure to Tab-Simco AMD. The metabolic shift of vetiver shoots was dramatic with longer-term AMD exposure, including upregulation of amino acid and glutathione metabolism, cellular respiration and photosynthesis pathways, with downregulation of phosphorylated metabolites. Meanwhile, the roots demonstrated drastic downregulation of phospholipids and phosphorylated metabolites, cellular respiration, glyoxylate metabolism, and amino acid metabolism. Vetiver accumulated ornithine and oxaloacetate in the shoots, which could function for nitrogen storage and various intracellular functions, respectively. Organic acids and glutathione were secreted from the roots for rhizospheric metal-chelation, whereas phosphorylated metabolites were recycled for phosphorus. These findings reveal AMD-induced metabolic shifts in vetiver grass, which are seemingly unique in comparison to independent abiotic stresses reported previously.

Efficacy and repellency of some essential oils and their blends against larval and adult house flies, Musca domestica L. (Diptera: Muscidae).

House flies are global pests and notoriously difficult to control. Essential oils of vetiver, cinnamon, and lavender and their blends were tested for toxic and repellent effects against larval and adult flies. All of the oils had moderate toxicity for eggs. Mortality of 2nd instar larvae was 57-78% in dipping assays, 38-100% in contact assays, and 94-100% in treated media. Lavender was less effective (38% mortality) than the others (91-100%) in contact bioassays. Oil blends were not more effective against larvae than individual oils. Vetiver and cinnamon oils were strongly repellent (84 and 78%, respectively) for larvae in treated media. None of the oils were repellent for adult house flies in olfactometer assays, but testing of additional products demonstrated significant repellency for neem oil, p-menthane-3,8-diol (PMD), and vanillin. Contact/fumigant toxicity of vetiver, cinnamon, and lavender oils was 100%, significantly higher than mortality from sunflower oil (67%). Blends of oils were not more effective against adults than the individual oils, but blends diluted with sunflower oil were as effective as the individual oils. Essentials oils of vetiver and cinnamon may have potential for fly management in situations where conventional insecticides cannot be used.

Bioguided identification of antimicrobial compounds from Chrysopogon zizaniodes (L.) Roberty root essential oil.

Aim: To determine the group of compounds from Chrysopogon zizaniodes root essential oil that have antimicrobial activity. Materials & methods: Thin-layer chromatography coupled to direct bioautography was used to determinate the fraction(s) having antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREF). Through GC-MS identification, the fractions with the greatest similarity to the active thin-layer chromatography fraction were used to determinate the MIC. Results: The subfraction 8 from column chromatography was responsible for the best MIC for MRSA (62.5 µg/ml) and VREF (125 µg/ml). Five compounds possibly responsible for antimicrobial activity were preliminary identified. Conclusion: We suggest that Cedr-8-en-13-ol, could be the more relevant compound involved in the antimicrobial activity in this study.

Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered E. coli.

The sesquiterpene (+)-zizaene is the direct precursor of khusimol, the main fragrant compound of the vetiver essential oil from Chrysopogon zizanioides and used in nearly 20% of men's fine perfumery. The biotechnological production of such fragrant sesquiterpenes is a promising alternative towards sustainability; nevertheless, product recovery from fermentation is one of the main constraints. In an effort to improve the (+)-zizaene recovery from a metabolically-engineered Escherichia coli, we developed an integrated bioprocess by coupling fermentation and (+)-zizaene recovery using adsorber extractants. Initially, (+)-zizaene volatilization was confirmed from cultivations with no extractants but application of liquid-liquid phase partitioning cultivation (LLPPC) improved (+)-zizaene recovery nearly 4-fold. Furthermore, solid-liquid phase partitioning cultivation (SLPPC) was evaluated by screening polymeric adsorbers, where Diaion HP20 reached the highest recovery. Bioprocess was scaled up to 2 L bioreactors and in situ recovery configurations integrated to fermentation were evaluated. External recovery configuration was performed with an expanded bed adsorption column and improved (+)-zizaene titers 2.5-fold higher than LLPPC. Moreover, internal recovery configuration (IRC) further enhanced the (+)-zizaene titers 2.2-fold, whereas adsorption velocity was determined as critical parameter for recovery efficiency. Consequently, IRC improved the (+)-zizaene titer 8.4-fold and productivity 3-fold from our last report, achieving a (+)-zizaene titer of 211.13 mg L-1 and productivity of 3.2 mg L-1 h-1. This study provides further knowledge for integration of terpene bioprocesses by in situ product recovery, which could be applied for many terpene studies towards the industrialization of fragrant molecules.

Mixed plantation of wheat and accumulators in arsenic contaminated plots: A novel way to reduce the uptake of arsenic in wheat and load on antioxidative defence of plant.

Wheat (W) and accumulators (A) were planted in plots (arsenic amended soil and without arsenic) designed with ecotoxicological concern for arsenic safe-grains. For the study sixteen plots of 2 × 2 × 0.5 m (l × b × h) size were prepared. Arsenic (As) in the form of sodium arsenate was applied at 50 mg/kg in plots. Out of these sixteen plots eight plots had arsenic amended soil and rest 8 without any arsenic (C). Accumulator's viz. Pteris vittata (PV), Phragmites australis (PA) and Vetiveria zizanioides (VZ) were planted along with wheat in combination (W + PV, W + PA and W + VZ) in twelve plots (6 AWAs plots and 6 AWC plots). In the rest 4 plots (2 WAs plots and 2 WC plots), only wheat was planted. The study was conducted for two cropping seasons, where accumulators were left in the plots between the cropping seasons except that before 2nd cropping accumulators were properly pruned and extra tillers were removed. The germination % of wheat in WAs in 1st and the 2nd cropping season was found to be 55 and 57%, while in AWAs and AWC plots it was between 86 and 92% (W + VZ, 56 and 73%). The physiological activity was found to be reduced in WAs plots compared to AWAs (except for vetiver combination) and AWC plots in both cropping seasons. The antioxidant activity was enhanced in WAs compared with AWAs. The arsenic concentration in grains of wheat was within the permissible limit set by WHO and GOI in AWAs plots while it exceeded the limit in W + VZ (in 1st cropping) and WAs in both cropings.

Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli.

The vetiver essential oil from Chrysopogon zizanioides contains fragrant sesquiterpenes used widely in the formulation of nearly 20% of men's cosmetics. The growing demand and issues in the supply have raised interest in the microbial production of the sesquiterpene khusimol, the main compound of the vetiver essential oil due to its woody smell. In this study, we engineered the biosynthetic pathway for the production of (+)-zizaene, the immediate precursor of khusimol. A systematic approach of metabolic engineering in Escherichia coli was applied to modulate the critical bottlenecks of the metabolic flux towards (+)-zizaene. Initially, production of (+)-zizaene was possible with the endogenous methylerythritol phosphate pathway and the codon-optimized zizaene synthase (ZS). Raising the precursor E,E-farnesyl diphosphate supply through the mevalonate pathway improved the (+)-zizaene titers 2.7-fold, although a limitation of the ZS supply was observed. To increase the ZS supply, distinct promoters were tested for the expression of the ZS gene, which augmented 7.2-fold in the (+)-zizaene titers. Final metabolic enhancement for the ZS supply by using a multi-plasmid strain harboring multiple copies of the ZS gene improved the (+)-zizaene titers 1.3-fold. The optimization of the fermentation conditions increased the (+)-zizaene titers 2.2-fold, achieving the highest (+)-zizaene titer of 25.09 mg L-1. This study provides an alternative strategy to enhance the terpene synthase supply for the engineering of isoprenoids. Moreover, it demonstrates the development of a novel microbial platform for the sustainable production of fragrant molecules for the cosmetic industry.

Phytoassessment of Vetiver grass enhanced with EDTA soil amendment grown in single and mixed heavy metal-contaminted soil.

Over the years, ethylene-diamine-tetra-acetate (EDTA) has been widely used for many purposes. However, there are inadequate phytoassessment studies conducted using EDTA in Vetiver grass. Hence, this study evaluates the phytoassessment (growth performance, accumulation trends, and proficiency of metal uptake) of Vetiver grass, Vetiveria zizanioides (Linn.) Nash in both single and mixed heavy metal (Cd, Pb, Cu, and Zn)-disodium EDTA-enhanced contaminated soil. The plant growth, metal accumulation, and overall efficiency of metal uptake by different plant parts (lower root, upper root, lower tiller, and upper tiller) were thoroughly examined. The relative growth performance, metal tolerance, and phytoassessment of heavy metal in roots and tillers of Vetiver grass were examined. Metals in plants were measured using the flame atomic absorption spectrometry (F-AAS) after acid digestion. The root-tiller (R/T) ratio, biological concentration factor (BCF), biological accumulation coefficient (BAC), tolerance index (TI), translocation factor (TF), and metal uptake efficacy were used to estimate the potential of metal accumulation and translocation in Vetiver grass. All accumulation of heavy metals were significantly higher (p < 0.05) in both lower and upper roots and tillers of Vetiver grass for Cd + Pb + Cu + Zn + EDTA treatments as compared with the control. The single Zn + EDTA treatment accumulated the highest overall total amount of Zn (8068 ± 407 mg/kg) while the highest accumulation for Cu (1977 ± 293 mg/kg) and Pb (1096 ± 75 mg/kg) were recorded in the mixed Cd + Pb + Cu + Zn + EDTA treatment, respectively. Generally, the overall heavy metal accumulation trends of Vetiver grass were in the order of Zn >>> Cu > Pb >> Cd for all treatments. Furthermore, both upper roots and tillers of Vetiver grass recorded high tendency of accumulation for appreciably greater amounts of all heavy metals, regardless of single and/or mixed metal treatments. Thus, Vetiver grass can be recommended as a potential phytoextractor for all types of heavy metals, whereby its tillers will act as the sink for heavy metal accumulation in the presence of EDTA for all treatments.

Chemical Composition, Antioxidant, and Antimicrobial Activities of Vetiveria zizanioides (L.) Nash Essential Oil Extracted by Carbon Dioxide Expanded Ethanol.

In the present study, the composition of essential oil isolated from the roots of Vetiveria zizanioides (L.) Nash, harvested in China, was studied, along with the bioactivities. A green novel method using an eco-friendly solvent, CO2-pressurized ethanol, or carbon dioxide expanded ethanol (CXE) was employed to isolate the essential oil from the root of Vetiveria zizanioides (L.) Nash with the purpose of replacing the traditional method and supercritical fluid extraction (SFE). After investigating the major operating factors of CXE, the optimal conditions were obtained as follows: 8.4 MPa, 50 °C, 5 mL/min ethanol, and 0.22 mole fraction of CO2, presenting an extraction oil that ranged from 5.12% to 7.42%, higher than that of hydrodistillation (HD) or indirect vapor distillation (IVD). The Gas Chromatography-Mass Spectrometry (GC-MS) analysis showed that three major components, including valerenol (18.48%), valerenal (10.21%), and ß-Cadinene (6.23%), are found in CXE oil, while a total of 23 components were identified, 48 components less than using conventional hydrodistillation. Furthermore, the antimicrobial activities of root oils were evaluated by the microdilution method, which showed that CXE oil exhibited an ability against Gram-positive bacteria, especially Staphylococcus aureus, approximately equivalent to traditional samples. Additionally, the DPPH free radical scavenging assay demonstrated that the antioxidant abilities of root oils were sorted in the descending order: IVD > HD > CXE > SFE. In conclusion, after a comprehensive comparison with the conventional methods, the CXE-related technique might be a promising green manufacturing pattern for the production of quality vetiver oil, due to the modification of ethanol by the variable addition of non-polar compressible CO2, ultimately resulting in a prominent dissolving capability for the extraction of vetiver solutes.

Arsenic Removal of Contaminated Soils by Phytoremediation of Vetiver Grass, Chara Algae and Water Hyacinth.

This research has been carried out for assessing phytoremediation of contaminated soils with 4 concentrations of arsenic by three plants, namely Vetiver grass (Vetiveria zizanioides), Chara algae (Chara vulgaris) and Water hyacinth (Hyacintus orientalis). The experimental results showed that at least two sampling times were significantly different. In addition, at least two plants were also significantly different in terms of percentages of total arsenic that were removed from the soil of the pots, as well as significant interactions between plant and arsenic concentrations. The results obtained from the thermodynamic studies show that, obtained by zero Gibbs free-energy, the process reached an equilibrium on the 60th day of the experiment, and, in fact, the adsorption of arsenic after the 60th day would be negligible.

Potential of essential oils to prevent fly strike and their effects on the longevity of adult Lucilia sericata.

Lucilia sericata is a facultative ectoparasite causing fly strike or myiasis in warm-blooded vertebrates. It is controlled by traps or insecticides, but both have drawbacks and alternative ways of control are urgently needed. Essential oils (EOs) of vetiver (Chrysopogon zizanioides), cinnamon (Cinnamomum zeylanicum), and lavender (Lavandula angustifolia) and their blends (OBs); OB1 (2 ml of each EO plus 4 ml of sunflower oil as a carrier) and OB2 (2 ml of each EO) were tested. Oils were tested at 5% for deterrence assays, and a dose response assay 0.01-0.6%, was conducted to determine forced-contact toxicity. We evaluated the efficacy of oils as oviposition deterrents, repellents/attractants, and their effects on mortality and longevity of adult L. sericata. Our data indicated that 0.2% EOs killed all flies by 5 min post-treatment and that vetiver oil greatly deterred flies from the oviposition medium and reduced adult longevity. Sunflower oil repelled all flies from ovipositing and greatly reduced the lifespan of treated adults. The blend of the four oils (OB1) had the greatest repellent effect on the flies. EOs have insecticidal, repellent, and oviposition-deterrent activities against L. sericata that could be used for suppression of blow fly populations.

Wound Pathogens: Investigating Antimicrobial Activity of Commercial Essential Oil Combinations against Reference Strains.

This study aimed to determine the antimicrobial activity of 247 essential oil combinations against the reference strains of wound pathogens. Essential oil combinations were investigated for antimicrobial activity against five pathogens. The minimum inhibitory concentration (MIC) assay was used and the fractional inhibitory concentration index (ΣFIC) calculated to determine interactions between selected oils. Twenty-six combinations displayed broad-spectrum antimicrobial activity against all five reference strains and several displayed synergy against more than one pathogen. The combination of Santalum austrocaledonicum (sandalwood) with Commiphora myrrha (myrrh) displayed noteworthy antimicrobial activity against all five reference strains and synergy against four (MIC values 0.03-1.00 mg/ml and ΣFIC values 0.19-1.00 mg/ml) pathogens. No antagonism was observed. Santalum spp. and Vetiveria zizanioides essential oils contributed the most to antimicrobial activity in combination. Essential oil combinations are presented as a viable option in wound therapy.

Vetiver grass (Chrysopogon zizanioides) is capable of removing insensitive high explosives from munition industry wastewater.

Synthetic organic explosive compounds in the wastewater stream of industrial munition facilities are subject to regulatory permits and require pretreatment prior to discharge. Munition industries are currently focused on developing insensitive high explosives (IHEs) such as dinitroanisole (DNAN), nitroguanidine (NQ), and 1,2,4-triazol-3-one (NTO), to replace conventional munitions such as trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). IHEs are typically more soluble than conventional explosives, and their production generates waste streams with high nitrate (N) concentrations. Several chemical remediation studies have attempted to degrade the explosive compounds within waste streams with limited success. Phytoremediation is a relatively new application for the remediation of munition industry wastewater, which is both environmentally and economically sustainable. Vetiver grass (Chrysopogon zizanioides), with its massive and dense root system and ability to grow in harsh environments, has been observed to remove many chemicals from soil and water, including nutrients and TNT. The objective of this study was to evaluate the phytoremediation potential of vetiver in removing explosive compounds and N from wastewater effluents generated in an industrial munition facility. Results show that the removal efficiency of vetiver was a factor of the initial concentration. Successive batches of vetiver removed DNAN, NQ, and RDX by 96, 79 and 100%, respectively. More than 95% of N was removed by four successive batches of vetiver grass. A major portion of NQ and RDX was translocated from root to shoot. LC-MS analysis showed the presence of transformation products of RDX, HMX (1,3,5,7-Tetranitro-1,3,5,7-tetrazocane) and DNAN in vetiver root and shoot.

Comparative metabolic profiling of vetiver (Chrysopogon zizanioides) and maize (Zea mays) under lead stress.

Lead (Pb) contamination of residential soils in United States is attributed to use of Pb based paints prior to 1978 and their deterioration and accumulation in surface soils. Exposure to Pb due to ingestion and inhalation of Pb laden soil and dust causes neurological disorders, renal disorders, developmental and behavioral problems, particularly in children under the age of six. Vetiver grass is one of the leading choices for Pb remediation due to its ability to hyperaccumulate Pb, in addition to high biomass. In order to understand the effect of Pb on vetiver metabolic pathways, we compared the global metabolic changes in vetiver with that of maize, a Pb susceptible plant under Pb stress. Vetiver showed massive increase in levels of key metabolites in response to Pb, including amino acids, organic acids and coenzymes. Maize showed very modest increase in some of the same metabolites, and no change in others. The results provide the first indication of the difference in metabolic response of the hyperaccumulator, vetiver to lead stress as compared to maize.

In vitro comparison of three common essential oils mosquito repellents as inhibitors of the Ross River virus.

BACKGROUND: The essential oils of Cymbopogon citratus (CC), Pelargonium graveolens (PG) and Vetiveria zizanioides (VZ) are commonly used topically to prevent mosquito bites and thus the risk of infection by their vectored pathogens such as arboviruses. However, since mosquito bites are not fully prevented, the effect of these products on the level of viral infection remains unknown. OBJECTIVES: To evaluate in vitro the essentials oils from Reunion Island against one archetypal arbovirus, the Ross River virus (RRV), and investigate the viral cycle step that was impaired by these oils. METHODS: The essential oils were extracted by hydrodistillation and analyzed by a combination of GC-FID and GC×GC-TOF MS techniques. In vitro studies were performed on HEK293T cells to determine their cytotoxicity, their cytoprotective and virucidal capacities on RRV-T48 strain, and the level of their inhibitory effect on the viral replication and residual infectivity prior, during or following viral adsorption using the reporter virus RRV-renLuc. RESULTS: Each essential oil was characterized by an accurate quantification of their terpenoid content. PG yielded the least-toxic extract (CC50 > 1000 µg.mL-1). For the RRV-T48 strain, the monoterpene-rich CC and PG essential oils reduced the cytopathic effect but did not display virucidal activity. The time-of-addition assay using the gene reporter RRV-renLuc showed that the CC and PG essential oils significantly reduced viral replication and infectivity when applied prior, during and early after viral adsorption. Overall, no significant effect was observed for the low monoterpene-containing VZ essential oil. CONCLUSION: The inhibitory profiles of the three essential oils suggest the high value of the monoterpene-rich essential oils from CC and PG against RRV infection. Combined with their repellent activity, the antiviral activity of the essential oils of CC and PG may provide a new option to control arboviral infection.

Phytosynthesized silver nanoparticles as antiquorum sensing and antibiofilm agent against the nosocomial pathogen Serratia marcescens: an in vitro study.

AIM: Serratia marcescens is an important multidrug-resistant human pathogen. The pathogenicity of S. marcescens mainly depends on the quorum sensing (QS) mechanism, which regulates the virulence factors production and biofilm formation. Hence, targeting QS mechanism in S. marcescens will ultimately pave the way to combat its pathogenicity. Thus, the present study is intended to evaluate the efficacy of Vetiveria zizanioides root extract-mediated silver nanoparticles (AgNPs) as a potent anti-QS and antibiofilm agent against S. marcescens. METHODS AND RESULTS: The AgNPs were synthesized using V. zizanioides aqueous root extract and the physiochemical properties of V. zizanioides-based AgNPs (VzAgNPs) were evaluated using analytical techniques such as ultraviolet-visible absorption spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, dynamic light scattering and scanning and transmission electron microscopic techniques. VzAgNPs were found to attenuate the QS-dependent virulence factors, namely prodigiosin, protease, lipase, exopolysaccharide productions and biofilm formation of S. marcescens, without inhibiting its growth. Further, the transcriptomic analysis confirmed the down-regulation of QS-dependent genes, which encode for the production of virulence factors and biofilm formation. CONCLUSION: The current study confirms VzAgNPs as an ideal anti-QS and antibiofilm agent against S. marcescens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first approach that validates the anti-QS and antibiofilm potential of phytosynthesized VzAgNPs against the nosocomial pathogen, S. marcescens. As VzAgNPs exhibits potent antivirulent activities, it could be used to treat hospital-acquired S. marcescens infections.