Insecticidal and acetylcholinesterase inhibitory activities of Achillea biebersteinii essential oil and its nanoemulsion and major monoterpenes against Tribolium castaneum

Essential oil (EO) was hydrodistilled from Achillea biebersteinii and analyzed using gas chromatography–flame ionization detection (GC–FID) and (GC–MS). Three monoterpenes, α-terpinene, p-cymene, and camphor were isolated as the main terpenes in the EO. Oil-in-water nanoemulsion (particle size 52.7 ± 4.4 nm) was prepared and characterized from EO through a hydrothermal approach. The EO products caused insecticidal effects and altered F₁ progeny of the red flour beetle, Tribolium castaneum (Herbst) in a dose and exposure time-dependent manner. The nanoemulsion was superior in bioactivity, followed EO. At 12.5 µl/cm², nanoemulsion and EO caused 100% mortality of the second larvae after 4 days of exposure with reduction of F₁ progeny of 100.0 and 89.2%, respectively. At 50.0 µl/cm², percentage mortality and reduction in progeny of EO products ranged between (61.4–100%) and (53.0–100%), respectively. Upon fumigation, nanoemulsion at 10 µl/L air caused 100% mortality of the second larvae after 4 days of exposure. At 20 µl/L air and 4 days after treatment, percentage mortality with the remainder botanicals ranging between 64.7 and 100. LC₅₀′s of products after 48 h exposure ranging between 6.7 and 58.1 µg/cm² in insecticidal bioassay, and between 3.6 and 26.3 µl/L upon fumigation. Botanicals caused a remarkable inhibition of acetylcholinesterase activity (AChE), where nanoemulsion (IC₅₀ = 14.22 mM), EO (IC₅₀ = 35.12 mM) and camphor (IC₅₀ = 23.93 mM) were superior as AChE inhibitors. There is a potential for using of A. biebersteinii EO, its nanoemulsion and monoterpenes as natural grain protectants after the required toxicological investigations.     

Phenotypic profile of Aedes albopictus (Skuse) exposed to pyrethroid-based mat vaporizers and underlying detoxification mechanisms: A statewide report in Selangor,Malaysia

This study examines the biological efficacy of four mosquito mat vaporizers each containing different active ingredients: prallethrin with PBO, dimefluthrin, prallethrin, and d-allethrin. The glass chamber assay was used to evaluate their efficacy on Aedes albopictus (Skuse) (Diptera: Culicidae) from nine districts in Selangor, Malaysia. Aedes albopictus exhibited different knockdown rates, with 50% knockdown times, KT₅₀, varying from 1.19 to 2.00 min, 1.22 to 2.20 min, 1.39 to 5.85 min, and 1.39 to 1.92 min for prallethrin with PBO, dimefluthrin, prallethrin and d-allethrin, respectively. In general, all populations of Ae. albopictus were completely knocked down after exposure to all active ingredients except Hulu Selangor population, which showed 96.00% knockdown against d-allethrin. On the contrary, mortality rates were observed from 84.00–100.00%, 84.00–100.00%, 90.67–100.00% and 90.67–100.00% in populations tested with prallethrin with PBO, dimefluthrin, prallethrin and d-allethrin, respectively. Moreover, significant correlations between mortality rates of prallethrin with PBO vs dimefluthrin (r = 0.836, P = 0.003), prallethrin with PBO vs prallethrin (r = 0.760, P = 0.011), and prallethrin vs d-allethrin (r = 0.694, P = 0.026) were also observed, suggesting cross-resistance among pyrethroids. d-allethrin was found to be high in insecticidal activity, followed by prallethrin, prallethrin with PBO, and dimefluthrin. In consistent with mortality due to insecticide exposure, elevated levels of enzyme activities were also demonstrated in Sabak Bernam, Hulu Selangor, Gombak, Petaling, Hulu Langat and Klang populations.     

Essential oil of Bassia muricata: Chemical characterization,antioxidant activity,and allelopathic effect on the weed Chenopodium murale

The essential oil (EO) of Bassia muricata shoots was extracted via hydro-distillation and then investigated by gas chromatography-mass spectrometry. Thirty-four compounds were recognized for the first time from this plant, representing 100% of the total mass. Terpenoids represented the major components with 69.17% of the total mass, containing oxygenated sesquiterpenes (53.18%), oxygenated monoterpenes (9.77%), sesquiterpene hydrocarbons (5.03%), and diterpenes (1.19%). Additionaly, 6-methoxy-1-acetonaphthone was the only aromatic compound represented in a high percentage of the total identified compounds with 22.35%. Additionally, a percent of 8.48% of the total mass was hydrocarbons. Only one oxygenated sesquiterpene namely hexahydrofarnesyl acetone representing 47.35% of the total mass was identified. It was followed by methoxy-1-acetonaphthone (19.92%), n-dotriacontane (3.58%), endo-borneol (3.24%), 6-methy-α-ionone (3.04%), and α-gurjunene (2.65%). The EO exhibited moderate antioxidant activity comparable with ascorbic acid as a standard, where it attained IC₅₀ value of 20.70 µL L⁻¹ and 16.32 µL L⁻¹, for DPPH and ABTS. The EO of B. muricata significantly reduces the germination and seedling development of the weed Chenopodium murale. The EO showed an IC₅₀ value of 175.60 µL L⁻¹, 246.65 µL L⁻¹, and 308.33 µL L⁻¹ for root growth, shoot growth, and germination, respectively. Therefore, this EO could be a good green resource for the control of weeds.     

Bioactivity of essential oil of Litsea cubeba from China and its main compounds against two stored product insects

During our screening program for agrochemicals from Chinese medicinal herbs and wild plants, the essential oil of Litsea cubeba fruits was found to possess strong contact toxicity against the cigarette beetle Lasioderma serricorne adults and the booklouse Liposcelis bostrychophila, with LD₅₀ values of 27.33 μg/adult and 71.56 μg/cm², respectively, and also showed strong fumigant toxicity against the two stored product insects with LC₅₀ values of 22.97 and 0.73 mg/L, respectively. The essential oil obtained by hydrodistillation was investigated by GC MS. The main components of the essential oil were identified to be E-citral (geranial) (27.49%), Z-citral (neral) (23.57%) and d-limonene (18.82%) followed by β-thujene (3.34%), β-pinene (2.85%), α-pinene (2.57%), 6-methyl-5-hepten-2-one (2.40%) and linalool (2.36%). Citral (Z/E-citral), d-limonene, β-pinene, α-pinene and linalool were separated and purified by silica gel column chromatography and preparative thin layer chromatography, and further identified by means of physicochemical and spectrometric analysis. Citral and linalool showed strong contact toxicity against L. serricorne and L. bostrychophila (LD₅₀ = 11.76, 12.74 μg/adult and 20.15, 99.97 μg/cm², respectively) and fumigant toxicity against L. serricorne and L. bostrychophila (16.54, 18.04 mg/L air and 0.14, 0.71 mg/L air, respectively). Otherwise, citral, d-limonene and linalool were strongly repellent against the cigarette beetle L. serricorne as the essential oil whereas β-pinene and α-pinene exhibited weaker repellency against the cigarette beetle compared with the positive control, DEET. Moreover, except α-pinene and linalool, the other three compounds as well as the essential oil exhibited comparable repellency against the booklouse relative to DEET.     

Fumigant toxicity of Lavandula spica essential oil and linalool on different life stages of Tribolium confusum (Coleoptera: Tenebrionidae)

The confused flour beetle, Tribolium confusum, is a common and severe pest of stored products. Here, using fumigation tests during four different exposure times, we evaluated the toxicity of different doses of essential oil of spike lavender, Lavandula spica and one of its major constituents, linalool, on different life stages of T. confusum under laboratory conditions. The toxicity of the L. spica oil and linalool varied as a function of the developmental stage and treatment duration. Young larvae (L1) were the most susceptible to toxic effects, with LC₅₀ = 19.535 μl/L of air for L. spica oil and LC₅₀ = 14.198 μl/L of air for linalool after 24 h of exposure, whereas older larvae (L8) were affected only very little by fumigation. Linalool caused higher egg mortality than L. spica oil at equivalent doses, but lower mortality in pupae and adults. Emergence of intact adult insects from surviving eggs, larvae and pupae was further reduced as a function of dose and exposure time to both L. spica oil and linalool compared to control-treated insects. Our results show that L. spica oil and linalool might be suitable for biological control of T. confusum, but tests at a larger scale are necessary to confirm our results.     

Chemical composition and biological activities of Laurus essential oils from different Macaronesian Islands

Essential oils (EO) and eight pure components from the fresh leaves of Laurus novocanariensis (LN) from Madeira and of Laurus azorica (LA) from four Azorean Islands (S. Miguel-SMI, S. Maria-SMA, Pico-PIC and S. Jorge-SJO) were evaluated for fumigant and contact insecticidal effects on adult stage of Mediterranean fruit fly (medfly) and for inhibition of oviposition. The chemical composition of EO were analysed by GC/MS. Oxygen-containing mono- and sesquiterpenes (mainly 1,8-cineole) dominated in LN (50%), LA-SMA (88%) and LA-SMI (57%) and mono- and sesquiterpene hydrocarbons (mainly α-pinene) dominated in LA-PIC (61%) and LA-SJO (44%). Linalool and α-terpinyl acetate were also abundant in LA-SMA and trans-cinnamyl acetate in LA-SJO. These three components and all of the Laurus' EO showed high repulsive activity against medfly oviposition. In contact assays on medfly adults, a moderate degree of mortality was observed, being the most toxic samples, in decreasing order, trans-cinnamyl acetate > α-terpinyl acetate > LA-SMI ∼ LA-SMA > LA-SJO ∼ linalool. In fumigant assays, α-terpinyl acetate was the only compound that showed some toxicity on medfly adults. According to the obtained results, the Laurus EO seem promising to be used against medfly oviposition in integrated pest management strategy.     

Chemical Compositions,Antioxidant and Antimicrobial Activities of Essential Oils of Psidium guajava L. Leaves from Different Geographic Regions in China

Hydrodistilled essential oils (EO) of Psidium guajava L. leaves from different regions in China were analyzed by GC and GC/MS. The samples from Guangdong Province displayed high EO yields (0.61 – 0.75%, v/w). A total of 50 components, representing over 98.00% of the EOs, were identified and semi‐quantitatived. The major constituents of EOs included β‐caryophyllene (17.17 – 31.38%), γ‐gurjunene (9.17 – 15.22%), τ‐cadinol (1.35 – 10.02%) and calamenene (2.13 – 7.80%). The terpenoids in all sample oils were dominated by sesquiterpenes hydrocarbons (70.18 – 84.35%), followed by oxygenated sesquiterpenes (9.89 – 22.19%). The similarities and differences among EOs from different samples were evaluated by hierarchical cluster analysis and principal component analysis methods. The IC₅₀ values of EOs from different regions were between 18.52 – 33.72 mg/ml (DPPH) and 13.12 – 25.15 mg/ml (ABTS⁺). The FRAP value of EO from Guangdong Province was 7.34 – 9.13 mmol Vc/g DM, while the FRAP value of EO from Taiwan Province was 2.29 – 2.36 mmol Vc/g DM. The antimicrobial tests revealed that EO had a higher antimicrobial activity against all Gram‐positive bacteria and two fungi. Moreover, EO from P. guajava leaves of Guangdong Province showed the highest antimicrobial activity. These properties can be considered in the design of industrial products and for further application in the food, pharmaceutical and cosmetic industries.     

Japanese beetle lures used alone or combined with structurally related chemicals to trap NE China scarabs (Coleoptera: Scarabaeidae)

Scarab beetles are agriculturally important worldwide, and as adults or larvae they may cause damage to the leaves, flowers, fruit, and roots of crops. Previous international studies showed that Japanese beetle (Popillia japonica, Newman) lures, and structurally related chemicals, can attract numerous scarabs. Based on those studies, season-long trials in grape, cabbage, corn and soybean fields were conducted in 2012–2013 in NE China. Tests determined the attractiveness of the Japanese beetle floral lure (phenethyl propionate:eugenol:geraniol, 3:7:3) and sex attractant (Japonilure), alone, combined, or in a mixture with either (Z)-3-hexen-1-ol, anethole = 1-methoxy-4-propenyl benzene or benzyl alcohol, to local Coleoptera. Furthermore, control efficacies based on leaf, ear, and silk damage, as well as reductions of adults on plants, and overwintering larvae, were also determined. Eleven scarab species, and four non-scarab species, were captured. The addition of other chemicals increased the attractiveness of the two Japanese beetle lures to scarabs such as Potosia brevitarsis (Lewis), Oxycetonia jecunda Faldermann, Holotrichia diomphalia Bates et al., Popillia quadriguttata (Fabricius), Maladera verticalis (Fairmaire), and Metabolus impressifrons Fairmaire, and the chrysomelid beetle Chrysomela populi L. The floral lure, and floral lure plus Japonilure baited traps resulted in > 80% overwintering larvae and adult reductions in corn and cabbage fields, whereas the Japonilure traps gave similar results in the soybean fields. This indicates that the commercial Japanese beetle lure combination can be recommended for use by the Chinese farmers in the corn or soybean fields, and that the related chemicals can be used to increase the attractiveness of the Japanese beetle lures.     

Growth responses and essential oil profile of Salvia officinalis L. Influenced by water deficit and various nutrient sources in the greenhouse

Salvia officinalis L. is a medicinal plant extensively used in foods, traditional medicine, and the pharmacological industry. In the current study, the effects of different irrigation regimes [irrigation after 70 ± 5 (regular), 105 ± 5 (moderate drought stress), and 140 ± 5 (severe drought stress) mm evaporation] and nutrient sources (control, NPK, farmyard manure, foliar fertilizer, and hydrogel) were investigated on the growth parameters and essential oil (EO) components of S. officinalis in the greenhouse. The plants were harvested two times. The regular irrigation treatment had the most significant effect on plant height (51 cm), fresh and dry herb weight (51.5 and 18.1 g plant⁻¹), and fresh and dry leaf weight (40.1 and 13.1 g plant⁻¹). The highest amount of EO was observed after moderate drought stress (1.48%). The NPK treatment had the greatest effect on plant height (40 cm), branch number (19 per plant), fresh and dry herb weight (53.4 and 18.9 g plant⁻¹), fresh and dry leaf weight (41.2 and 13.6 g plant⁻¹), and EO content (1.67%). The 1^st cutting was superior in EO amount, while the 2^nd cutting had a high agronomic yield. α-Thujone (from 21.6 to 34.2%) was identified as the predominant compound. Additionally, the content of α-thujone in the 2^nd cutting was higher after moderate drought stress, NPK, and hydrogel treatments. Moreover, 1,8-cineole, β-thujone, camphene, α-pinene, α-humulene, viridiflorol, borneol, and bornyl acetate were the other main compounds. As a general result, regular irrigation and NPK treatments improved the agronomic yield of S. officinalis. The plants under drought stress produced high amounts of EO. The farmyard manure also improved plant yield by providing a part of the plant's nutritional needs. Therefore, it could be concluded that it is crucial to determine the effects of limited water availability and various nutrient sources on yield and chemical compositions for medicinal and aromatic plant growth.     

Piperonyl butoxide synergizes the larvicidal activity of Origanum vulgare essential oil and its major constituents against the larvae of Aedes albopictus and Culex pipiens quinquefasciatus

Mosquitoes bite human beings and transmit many diseases, such as malaria, dengue fever, and Zika virus. Vector control of mosquitoes is an effective strategy for reducing the spread of disease. However, extensive use of insecticides (e.g. pyrethroids and organophosphorus) has caused resistance in mosquitoes, which weakens the effectiveness of mosquito control. Phytochemicals have been considered an alternative approach to mosquito control. Essential oil (EO) was obtained from the leaves and flowers of Origanum vulgare, and its synergistic activity with piperonyl butoxide (PBO) was tested against Aedes albopictus and Culex pipiens quinquefasciatus larvae. Thirty-seven compounds were identified, among which carvacrol and thymol were two major constituents (30.73 % and 18.81 %, respectively). O. vulgare EO had a significant toxic effect against fourth-stage larvae of Cx. p. quinquefasciatus and Ae. albopictus, with LC₅₀ values of 17.51 and 75.90 mg/L. Carvacrol and thymol also each appeared to be more effective against Cx. p. quinquefasciatus (LC₅₀ = 19.30 and 11.56 mg/L, respectively) than Ae. albopictus (LC₅₀ = 26.62 and 26.66 mg/L, respectively). PBO interacted synergistically with O. vulgare EO, carvacrol and thymol with 2.60–6.26 times as much of the active compound needed without PBO as against Cx. p. quinquefasciatus and Ae. albopictus larvae. Overall, our results contribute to the development of new natural mosquito insecticides.     

Efficacy of Iranian diatomaceous earth deposits against Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae)

Laboratory bioassays were carried out to evaluate the insecticidal efficacy of two Iranian deposits of diatomaceous earths (DEs) and a commercial formulation, SilicoSec®, against adult confused flour beetle (Tribolium confusum Jacquelin du Val). The Iranian DEs were dried and sieved to get particle sizes of 0–149, 74–149, 0–74 μm, and 0–37 μm. First, DE samples were applied at the four concentration levels of 500, 1000, 1500 and 2000 ppm and each concentration was replicated four times. Tests were carried out at 27 ± 1 °C and 55 ± 5% r.h. in continuous darkness. The beetle mortality was counted at 2, 7, and 14 days after exposure. Moreover, another experiment was conducted to estimate the LC₅₀ values of the DEs. For the first experiment, adult beetle mortality exceeded 51% when exposed for 2 d to 2000 ppm of SilicoSec®. Complete mortality was recorded at each concentration of SilicoSec® at an exposure longer than 7 days except for 500 ppm; while mortalities of T. confusum at 2000 ppm of Maragheh and Mamaghan samples with the 0–149 μm particle size were 40.62% and 85.41%, respectively. Mortality of T. confusum was influenced by concentration and time of exposure to the DEs. SilicoSec® was the most effective DE followed by Mamaghan samples. The Maragheh samples were the least effective. In addition, in most cases fractions with smaller particles were more effective than larger ones. More experiments are necessary to process natural DEs and make them commercially exploitable.     

Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control

Lavandula angustifolia is a well known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of L. angustifolia essential oil (EO) against engorged Rhipicephalus (Boophilus) annulatus (Acari; Ixodidae) females. For this purpose six serial concentrations (0.5, 1.0, 2.0, 4.0, 6.0 and 8.0% w/v) of L. angustifolia EO were used. There was considerable mortality in concentrations more than 4.0% although there were no differences between 6.0 and 8.0% in all measured criteria. The mortality rate 24 h after inoculation was 73.26 and 100% in groups treated with 4.0 and 8.0% EO, respectively. Lavender EO also reduced tick egg weight in a concentration-dependent manner. The amount of eggs produced varied from 0.12 g (at 0.5% EO) to 0.00 g (at 8.0% EO) but did not differ statistically from the control. L. angustifolia EO caused 100% failure in egg laying at 6.0 and 8.0% whereas this value in the control group was zero. A positive correlation between L. angustifolia EO concentration and tick control, assessed by relative mortality rate and egg-laying weight, was observed by the EO LC/EC₅₀, which, when calculated using the Probit test, was 2.76-fold higher than the control. Lavender is a promising acaricidal against R. (B.) annulatusin vitro.     

Effects of variety and growth year on the essential oil properties of lavender (Lavandula angustifolia Mill.) and lavandin (Lavandula x intermedia Emeric ex Loisel.)

Essential oil (EO) properties of six L. angustifolia (LA) and two L. x intermedia (LI) cultivars were evaluated during summer harvest periods of 2017 and 2018, The cultivars were grown in Dörgicse, Hungary. According to our results, the essential oil (EO) content was highly dependent on the varieties in both experimental years (2017: p < 0.0001 and 2018: p < 0.0001). In 2017, the EO content values of LI accessions were all higher than those of the LA cultivars. However, in 2018, the mean EO content value of ‘Budakalászi’ (LA) (8.2 mL/100g) exceeded that of ‘Grosso’ (LI) (7.7 mL/100g). The EO of ‘Mailette’ (LA) and ‘Budakalászi’ (LA) contained the highest linalool proportions (52.9% and 47.0%) while ‘Beate’ (LA) showed the lowest (18.1%); and this latter variety was richest in linalyl-acetate (58.9%). LI varieties produced lower linalyl acetate and total ester ratios in comparison to the LA cultivars. Concerning EO composition, only the UK varieties ‘Hidcote Blue’ (LA) and ‘Munstead’ (LA) met the requirements of the Ph. Eur.This study provided evidence that the effect of growth year on the EO content was significant only in two varieties out of the eight. However, in the more rainy year 2018, the accumulation of linalool and linalyl acetate was significantly affected in several cases, where linalool proportions decreased and linalyl acetate ratios increased.     

In vivo and in vitro ethylene oxide exposure of human lymphocytes assessed by chemical stimulation of unscheduled DNA synthesis

Factory workers exposed to ethylene oxide (EO), 0.5–1.0 ppm in factory air, together with matched controls from the same factory, were examined for evidence of toxic exposure by measurement of unscheduled DNA synthesis (UDS) induced by N-acetoxy-2-acetylaminofluorene (NA-AAF) and of chromosome aberrations in peripheral lymphocytes.The total chromatid gaps plus breaks were significantly elevated and NA-AAF-induced UDS was significantly reduced in the EO-exposed group as compared with the unexposed control group. The NA-AAF-induced UDS values negatively correlated to the duration (yr) of EO exposure (r = ?0.45, p < 0.02) and the number of chromosome breaks (r = ?0.61, p < 0.05), indicating an inhibition in vivo of DNA-repair capacity by EO. These data were verified in vitro by biochemical and autoradiographic studies of EO-induced UDS in human blood cells. Above 2 mM EO, UDS was inhibited in lymphocytes whether they were cultured for 24 or 122 h after alkylation with EO. Even at the subtoxic EO dose of 0.1 mM, lymphocytes were sensitized to additional exposures of NA-AAF, so that cytotoxicity was increased to 40% compared with 5% for the controls even though UDS was unaffected.It is concluded that EO was toxic to lymphocytes, even when they were sensitized at non-toxic EO doses to the cytotoxic action of other mutagens (e.g. NA-AAF), and the cells that did survive above 2 mM EO were inhibited in their DNA-repair capacity as judged by reduced UDS.     

Development of a process for the production of nutrient supplements for fermentations based on fungal autolysis

This study verifies the potential of fungal autolysis as an alternative process for the production of nutrient-rich solutions similar to yeast extracts. Autolytic experiments were carried out on fermentation solids derived from either batch or continuous submerged cultivations of Aspergillus awamori on various wheat flour milling streams. The degree of autolysis was not affected by the pH range used (3–6.5), whereas it was severely affected by temperature (30–55 °C), initial solids concentration (10–45 g/L) and incubation time. The enzymatic disruption of the fungal cell wall was identified by image analysis as well as by the reduction in total dry weight and the gradual release of various components, such as free amino nitrogen and phosphorus. The novel method of autolysate recycling enabled the enrichment of the solution with lytic enzymes leading to increased fungal cell degradation rates. In this way, it was made possible to reduce the initial total dry weight by 47% and produce a nutrient-rich solution containing 1.6 g/L free amino nitrogen, 5.3 g/L total nitrogen and 0.5 g/L phosphorus.     

Antileishmanial activity of essential oil from Chenopodium ambrosioides and its main components against experimental cutaneous leishmaniasis in BALB/c mice

Chenopodium ambrosioides have been used during centuries by native people to treat parasitic diseases.Aims of the studyTo compare the in vivo anti-leishmanial activity of the essential oil (EO) from C. ambrosioides and its major components (ascaridole, carvacrol and caryophyllene oxide).Materials and methodsAnti-leishmanial effect was evaluated in BALB/c mice infected with Leishmania amazonensis and treated with the EO, main compounds and artificial mix of pure components by intralesional route at 30 mg/kg every 4 days during 14 days. Diseases progression and parasite burden in infected tissues were determined.ResultsEO prevented lesion development compared (p < 0.05) with untreated animals and treated with vehicle. In addition, the efficacy of EO was also statistically superior (p < 0.05) compared with the glucantime-treated animals. No potential effects were observed with pure components treatment. Mix of pure compounds cause death of animals after 3 days of treatment.ConclusionsOur results demonstrate the superiority of EO against experimental cutaneous leishmaniasis caused by L. amazonensis.     

Variation in Essential Oil Yield and Composition of Dorema aucheri Boiss., an Endemic Medicinal Plant Collected from Wild Populations in Natural Habitats

In the present research, variability in essential oil (EO) composition of five Dorema aucheri populations collected from natural habitats in different regions of Iran, were investigated. The EO content of populations varied from 0.28 to 0.68%. According to gas chromatography/mass spectrometry analysis, β‐caryophyllene (7.17 – 35.73%), thymol (23.45 – 29.64%), β‐gurjunene (2.58 – 5.89%), carvacrol (1.32 – 2.67%) and cuparene (1.97 – 2.98%) were the major components. Hierarchical cluster, principal component and canonical correspondence analyses classified the studied populations into three groups based on major EO components. The environmental parameters of the collected sites were also evaluated. According to the results, it might be suggested that sandy soils with high mean annual precipitation were major environmental factors influencing the amount of β‐caryophyllene, while thymol, cuparene and caryophyllen oxide increased in silty and clay soils. Finally, the population collected in high altitudes and clay soils had higher amount of β‐gurjunene.     

Toxicity and repellency of origanum essential oil and its components against Tribolium castaneum (Coleoptera: Tenebrionidae) adults

The components of Origanum vulgare L. essential oil showing insecticidal activity and repellency against red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), adults were analysed by GC-MS. All constituents were identified, and the main components were carvacrol (67.2%), p-cymene (16.2%), γ-terpinene (5.5%), thymol (4.9%), and linalool (2.1%). In a vapor phase fumigant assay, the origanum oil was more effective in closed conditions (LD₅₀ = 0.055 mg/cm³) than in open conditions (LD₅₀ > 0.353 mg/cm³). This suggests that toxicity is exerted largely in the vapor phase. Based on 24-h LD₅₀ values, the toxicity of caryophyllene oxide (0.00018 mg/cm³) was comparable with that of dichlorvos (0.00007 mg/cm³). In addition, thymol, camphene, α-pinene, p-cymene, and γ-terpinene showed good insecticidal activity (LD₅₀ = 0.012–0.195 mg/cm³). In repellency tests using 9 constituents of origanum oil, caryophyllene oxide showed complete repellency at 0.03 mg/cm². Hydrogenated monoterpenoids, such as thymol, α-pinene, carvacrol, and myrcene, elicited strong repellency at 0.03 and 0.006 mg/cm². Repellency depended on both time and concentration. These results indicate that origanum oil and its components could be potential candidates as a fumigant and repellent for managing T. castaneum adults.     

Simultaneous determination of nicotinic acid and its four metabolites in rat plasma using high performance liquid chromatography with tandem mass spectrometric detection (LC/MS/MS)

A sensitive and specific liquid chromatography electrospray ionization–tandem mass spectrometry method for the simultaneous quantitation of nicotinic acid (NicA) and its metabolites nicotinamide (NA), 1-methylnicotinamide (MNA), 1-methyl-2-pyridone-5-carboxamide (M2PY) and 1-methyl-4-pyridone-5-carboxamide (M4PY) in rat plasma has been developed and validated. As an internal standard, 6-chloronicotinamide was used. The samples (100 μL) were subjected to deproteinization with acetonitrile (200 μL) and then, after centrifugation, 150 μL of the supernatant was transferred into conical vial and evaporated. Dry residue was reconstituted in 100 μL of the ACN/water (10:90, v/v) mixture. Chromatography was performed on a Waters Spherisorb^® 5 μm CNRP 4.6 × 150 mm analytical column with gradient elution using a mobile phase containing acetonitrile and water with 0.1% of formic acid. The full separation of all compounds was achieved within 15 min of analysis. Detection was performed by an Applied Biosystems MDS Sciex API 2000 triple quadrupole mass spectrometer set at unit resolution. The mass spectrometer was operated in the selected reactions monitoring mode (SRM), monitoring the transition of the protonated molecular ions m/z 153–110 for M2PY, 153–136 for M4PY, 124–80 for NicA, 123–80 for NA and 137–94 for MNA. The mass spectrometric conditions were optimized for each compound by continuously infusing the standard solution at the rate of 5 μL/min using a Harvard infusion pump. Electrospray ionization (ESI) was used for ion production. The instrument was coupled to an Agilent 1100 LC system. The precision and accuracy for both intra- and inter-day determination of all analytes ranged from 1.3% to 13.3% and from 94.43% to 110.88%. No significant matrix effect (ME) was observed. Stability of compounds was established in a battery of stability studies, i.e. bench-top, autosampler and long-term storage stability as well as freeze/thaw cycles. The method proved to be suitable for various applications. In particular using this method we detected increased concentration of MNA and its metabolites in rat plasma after treatment with exogenous MNA (100 mg/kg), as well as increased concentration of endogenous NA and MNA in rat plasma in the early phase of hypertriglyceridemia development in rats fed high-fructose diet.     

Biomarkers of iron status and trace elements in welders

Iron status was studied in 137 welders exposed to a geometric mean (GM) air concentration of 214 μg/m³ (range 1–3230) of manganese (Mn), in 137 referents and in 34 former welders. The GM concentrations of S-ferritin were 119 (3–1498), 112 (9–1277) and 98 (12–989) μg/L (p = 0.24) in the three groups, respectively. Also the GM concentrations of S-hepcidin were not significantly different between the groups (8.4 μg/L (2.8–117); 6.6 μg/L (1.8–100); 6.5 μg/L (1.2–22)) (p = 0.22). Multiple linear regression analysis including all welders and referents showed an increase in the concentration of S-ferritin associated with having serum carbohydrate deficient transferrin (S-CDT) above the upper reference limit of ≥1.7%, indicating high alcohol consumption. Serum C-reactive protein was not associated with exposure as welders, but an association with S-ferritin was shown. The GM S-ferritin concentrations among all welders and referents with S-CDT ≥ 1.7% were 157 μg/L (95% CI 113–218) as compared to 104 μg/L (95% CI 94–116) (p = 0.02) in those with S-CDT < 1.7%. The GM concentrations of Mn in biological fluids were higher in the welders as compared to the referents, while S-Fe, S-Co and B-Co were statistically significantly lower. This could suggest a competitive inhibition from Mn on the uptake of Fe and Co. Increasing concentrations of S-CDT was associated with higher S-Mn, S-Fe and B-Co in the multiple linear regression analysis. The association between S-CDT and S-Fe remained when all subjects with high S-CDT (≥1.7%) were excluded, suggesting increased uptake of Fe even at lower alcohol consumption.