Aloysiatriphylla is an aromatic plant used in several industrial sectors, owing to thechemical properties of its essentialoil. Biosynthesis of organic compounds is influenced by the growth conditions. As such, temporal and spatial variation affect thechemicalcompositionofessential oils. We hypothesized that: i) thechemicalcompositionof A. triphyllaessentialoil is influenced by differences in irrigationandseason; and ii) the major components of A. triphyllaessentialoil do not vary between treatment conditions. To test these hypotheses, we determined thechemicalcompositionof A. triphyllaessentialoil as a function of four seasons crossed with four irrigation levels. A completely randomized experimental design with a randomized block in a 4 × 4 factorial scheme, representing the four seasons ofthe year (summer, autumn, winter, and spring) and four irrigation levels (50%, 75%, 100%, and 125% ofthe reference evapotranspiration), was used with four replicates. Our results show that thechemicalcompositionof A. triphyllaessentialoil varied with the two study factors, of which season was the major factor. The highest concentrations of constituents classified as monoterpene and sesquiterpene were observed in the summer season, andthe highest constituents ofthe other group of compounds were observed in winter. In addition, the major components oftheessentialoil were α -citral, limonene, and β -citral, and their levels were the highest during winter.
The present study demonstrated that A. polystachya EO has an anesthetic effecton grouper juveniles. This EO induced sedation at all concentrations tested and anesthetized animals within 3 and 1.5 min (300 and 400 µL L −1 , respectively). Recovery time for both concentrations was about 8 min and 5 min, respectively, and no mortality was observed as a result of anesthesia induction. These findings are in accordance with literature criteria (Marking and Meyer 1985, Gilderhus and Marking 1987, Keene et al. 1998, Park et al. 2009). Parodi et al. (2014) tested the anesthetic effectofthe EO obtained from another species ofAloysia, namely A. triphylla, in concentrations ranging between 20 and 800 µL L −1 on two strains (albino and gray) of silver catfish (Rhamdia quelen), and 200 µL L −1 EO was the best concentration to induce anesthesia in the albino strain, while for the gray strain it was 400 µL L −1 EO. An emulsified mixture composed of Mentha spicata EO and methyl salicylate oil (containing 28.4% L-carvone) anesthetized common carp (Cyprinus carpio) within the recommended time at 395 µL L −1 (Roohi and Imanpoor 2014) and Atlantic salmon (Salmo salar) at 257 µL L −1 (Danner et al. 2011), but M. spicata EO alone induced anesthesia in less than 3 min only at 5000 µL L −1 (Roohi and Imanpoor 2015). Interestingly, increased EO concentration promoted recovery time decrease. Other EOs produced higher recovery times as the EO concentration increased (Cunha et al. 2010, Heldwein et al. 2012, Silva et al. 2012, 2013, Parodi et al. 2014). Since EOs are complex mixtures of compounds, a particular biological activity such as induction and recovery times from anesthesia depends onthe specific chemical characteristics of each EO, including the qualitative compositionand
The control of tick species that affect animal production is vital for the economic welfare ofthe cattle industry. This study focused on testing the acaricidal activity oftheessentialoil from the leaves and stems of Tagetes minuta against several Brazilian tick species, including Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, Amblyomma cajennense and Argas miniatus. Thechemicalcompositionoftheessentialoil was determined by chromatography and spectroscopy analyses, which revealed the presence of monoterpenes. The adult immersion test (AIT) andthe larval packet test (LPT) were used to evaluate the efficacy of T. minuta essentialoil in tick management at concentrations of 2.5, 5, 10, 20 and 40%. The results demonstrated that the T. minuta essentialoil had over 95% efficacy against four species of ticks at a concentration of 20%. These results suggest that theessentialoilof T. minuta could be used as an environmentally friendly acaricide.
Ocimum sanctum L. (Lamiaceae) sin. Ocimum tenuiflorum L. or Tulsi basil is a plant originating from the tropical and subtropical areas of India. It is used in both the traditional and official medicine in India. Tulsi is a type of basil that is insuffi- ciently explored and studied in Europe. The goal of this paper is to determine thechemicalcomposition, antioxidative, and antimicrobial activity oftheessentialoil Ocimum sanctum L. grown in Serbia. The quantity ofessentialoil in 100 g of herb (v/w) is 0.68%, with 41 components identified in the tested essentialoil. The most represented chemical group were sesquiturpene hydrocarbonates with 80.47%. Other groups were much less represented. Sesquiturpene hydrocarbonate β -car- iophyllene is a predominant component in theessentialoil with 63.80%. The quantity of tested essentialoil needed to achieve 50% of inhibition of DPPH radi- cals is 0.35 μg/ml, and it has high potential to neutralize free radicals. The essen- tial oil exhibited antibacterial activity to all tested strains of bacteria, both Gram- positive and Gram-negative. It affected all strains in an inhibitory way in the inter- val 0.34–41.50 μl/ml, and in a bactericide way within the range 22.50–124.5 μl/ml. The most sensitive strains of bacteria were Salmonella typhimurium and Escherichia coli, while Listeria monocytogenes and Enterococus faecalis showed greatest resistance. Theessentialoil exhibited antifungal activity on all tested fungi. It affected all tested fungi in an inhibitory way in the interval 4.42–8.83 μl/ml, and in a microbicide way within the range 10.00–50.00 μl/ml. The most sensitive fungi are: Aspergillus ochraceus, Penicillium ochrochloron and Penicilium funiculosum, while the most resistant one is Aspergillus niger. The tested basil essentialoil Oci- mum sanctum demonstrated significant antioxidative and antimicrobial effectand may be used as a raw material in food, pharmaceutical andchemical industries.
related to the antioxidant capacity and inhibition of lipid peroxidation shown by EOAT when used as an anesthetic in vivo in silver catfish (GRESSLER et al., 2012). Exposure in vivo to EOAT immediately before slaughter reduced stress in the catfish by suppressing cortisol release and increasing the activity of antioxidant enzymes, catalase and glutathione S-transferase (GRESSLER et al., 2012). In addition, the antioxidant effectof EOAT may also be related to the removal of free radicals by antioxidant compounds present in theoil, since, in vitro, theoil presented moderate capacity to remove the DPPH radical when compared to BHA (ALI et al., 2011). Despite presenting a promising antioxidant effectonthe frozen fillets and even though EOAT is a natural compound extracted from a plant that is widely used in tea and for spice in the human diet, it is still necessary to evaluate its toxicological safety before it can be used in foods.
As expected, the few studies on mint irrigation report that plant growth andessentialoil yield andcomposition were influenced by different levels ofirrigation regimes, following various water deficit stress levels . Okwany et al.  reported unchanged yield across apparently different irrigation treatments at the first cutting but a decrease at the driest plots during the second harvest due to the loss of leaf area index, as the biomass (hay) production is highly sensitive to water stress. Our data confirm their results on an increasing effectofirrigationon hay andoil yield per hectare, while our findings are different for the harvest dates, lower in our case for the second harvest. Our data also confirm the results of Nakawuka et al.  at Washington State University’s Irrigated Agriculture Research and Extension Center (IAREC), although hay andoil produced are lower here than in the IAREC study. Our results are also parallel to the results obtained by Khorasaninejad et al. , who indicated that drought stress motivated a significant reduction in all ofthe growth parameters andessentialoil yield and percentage. The highest values of growth parameters andessentialoil percent and yield were observed under full irrigation. In contrast, Hassanpour et al.  claims that the seedlings at moderate stress (treatment named 75 % FC) showed maximum growth and it was the threshold of drought-initiated negative effects on seedling growth. Oil yields of different M. longifolia L. genotypes were higher when mint was fully irrigated, in comparison with the stress treatment, named 60% FC.
Factors related to medicinal plant growing like plant spacing, cutting height, seson, age and harvest time may influence directly in theessentialoil production and, therefore, in its pharmacological properties. Some studies state the influence of these factors in aromatic species like Cymbopogon citratus (Nascimento et al., 2003), Lippia alba (Santos et al., 2004), Cymbopogon winterianus (Blank et al., 2007) andAloysiatriphylla (Brant et al., 2009). However, studies on plants of Zingiberaceae family are still rare. Phytochemical studies have advanced considerably, but the same is not true for agronomic studies. That is why the researches related to the agronomic aspects became essential (Innecco et al., 2003) to guarantee a regular supply of a great amount and quality of raw material for industries.
The decrease in the absorption at 560 nm suggested superoxide radicals’ scavenging, being most significant when theessentialoil was added to the reaction mixture – 95.93 %. Superoxyde dismutase 100 U/mL with inhibiting effect 77.8 % was used as a standard. The results achieved in the study revealed two ways of influence ofessentialoiland eugenol on xanthine-xanthine oxidase function – they acted as xanthine oxidase inhibitors and superoxide scavengers, thus confirming the interpretation of (Jubert et al., 2004).The evaluation of antioxidant activity in a linoleic acid model system was as follows: For the task of evaluating the inhibitory effectofessentialoilon lipid peroxidation, a model system of linoleic acid emulsion was applied. The antioxidant capacity was estimated both at the early stages of linoleic acid autoxidation and later, after the emergence of secondary oxidation products, expressed as malonic aldehyde content. Two indicators were referred to, corresponding to a different degree of lipid peroxidation – conjugated diene formation and TBARS. Theeffectoftheessential Murraya koenigii leaf oilon lipid peroxidation was assayed at body temperature, 37°C.
Aloysiatriphylla (12) and Cordia verbenacea DC (5), have been obtained and studied, this study had not yet been carried out on Lithraea molleoides (Vell.) Engler. The aims ofthe present study were: (i) the extraction oftheessentialoilofthe mature fruits, leaves and aerial parts ofthe plant and quantification ofthe yield thereof; (ii) the identification and quantification ofthe principal components oftheessentialoiland (iii) the determination of its antimicrobial activity against bacteria and yeast.
This study evaluated theeffectof salt stress onthe growth of Lippia gracilis Schauer, a species native to the caatinga (shrublands) of Brazil and rich in essential oils, as well as onthe quality of its oil. We exposed individuals of L. gracilis to NaCl, in the following concentrations, for a period of 40 days: 25 mM; 50 mM; 75 mM; and 100 mM. An additional group of plants was not exposed to NaCl (controls). Data were collected on days 20 and 40. We evaluated relative growth rate; shoot and root dry weight; relative water content; proline concentration in leaves; andchemicalcompositionoftheessentialoil. At all concentrations, NaCl reduced the relative growth rate in comparison with that observed for the controls. No significant difference in relative water content was observed among treatments. In all treatments, the proline concentration in leaves was highest on day 40. Salt stress did not affect the yield or the concentrations ofthe constituents oftheessentialoilof L. gracilis, carvacrol and thymol showing the highest concentrations in all treatments.
Antimicrobial susceptibility tests with plant extracts using the microdilution plate may have drawbacks such as precipitation of some components in the extract and adherence of some microorganisms to the bottom ofthe hole making it difficult to analyze (Ostrosky et al. 2008). However, liquid medium was considered the best medium for the evaluation of antimicrobial activity using the natural compounds (Skandamis et al. 2000). Methanol was used to facilitate the dissolution ofthe extracts in a liquid culture medium and assess its minimum bactericidal activity (data not shown). In the present work, methanol showed no antimicrobial activity against Aeromonas. The lack of antimicrobial activity of methanol had also been found in Gram-negative bacteria (Pinto et al. 2001). Celiktas et al (2007) found low antimicrobial activity of methanol for the genus Staphylococcus and this activity was considered insignificant when compared to theessentialoil tested.
TheessentialoilofAloysiatriphylla (L’Herit) Britton (EOA) presented anesthetic efficacy in the freshwater silver catfish Rhamdia quelen (PARODI et al., 2014), but no studies in other fish species have been performed. The fat snook, Centropomus parallelus, is an inshore tropical euryhaline species with high flesh quality and market value, as well as significant aquaculture potential (BOUCHEREAU et al., 2000). Therefore the aim of this study was to investigate the anesthetic induction and recovery times ofthe EOA, its efficacy as a stress-reducing agent during the transport of fat snook andthe influence of salinity ontheeffectof EOA.
EOVAC showed a rapid analgesic effect in the tail immersion test. The analgesic effectof EOVAC was started from 15 min after administration. This result suggested that EOVAC had a rapid analgesic effect. This rapid effectof EOVAC may be due to its lipophilic nature. Because of this lipophilic character, the EOVAC allowed to rapidly absorb from injection site and also rapidly penetrated the blood brain barrier and reached the central nervous system (Buchbauer et al., 1993). Some of constituents ofessential oils such as linalool, eugenol, menthol, and thymol can act onthe central and peripheral nervous system as ion channel modulators (De Araújo et al., 2011).
EFFECTOF DRYING-AIR TEMPERATURE ON CONTENT ANDCHEMICALCOMPOSITIONOFTHEESSENTIALOILOF Pectis brevipedunculata. Leaves and lower heads of P. brevipedunculata were submitted to four drying-air temperatures (room temperature, 40, 50 and 60 °C). Room temperature (approximately 30 °C) and higher temperature drying (50 and 60 °C) had a deleterious effectontheessentialoil content. The recommended drying-air temperature for the species is 40 °C for it results in the same amount ofessentialoil observed in fresh cut plants. Overall, 13 components accounting for more than 92% ofthe total composition were identiied. Citral was the major component, followed by α-pinene and limonene. Theessentialoil showed high toxicity against Artemia salina larvae.
Alpinia zerumbet is a medicinal plant from Asian origin used in folk medicine for the treatment of hypertension, which effect is attributed to terpinen-4-ol, the major component oftheessentialoil. The objective of this work was to identify theessentialoil secretory structures in the leaf, fl ower, root and rhizome of this plant, and analyze the content andthechemicalcompositionoftheoil in the different organs ofthe plant. Sections were subjected to histochemical test with Nadi reagent for in situ localization of secretion. Theessentialoil extraction was performed by hydrodistillation in a Clevenger apparatus andthe compounds were identifi ed in CG-EM/FID. The histochemical test was positive for terpenoids, confi rming the presence ofessentialoil stored in secretory structures named oils cells present in all analyzed organs. The higher essentialoil content was found onthe leaf (0.30%), while the petal andthe rhizome presented content of 0.10% and 0.06%, respectively. It was not possible to determine essentialoil content ofthe root due to the low amount of biomass produced. There were qualitative and quantitative differences in thechemicalcompositionoftheessentialoil in the different plant organs, but the major constituent in all of them was the terpinen-4- ol, followed by 1,8 cineol in the leaf and by the -terpineol in the fl ower and rhizome.
To sum up, results of this study showed that theessentialoil from Protium ovatum leaves found in the Brazilian Cerrado, which is located in the country’s central-west region, has promising antiparasitic potential with moderate cytotoxicity towards LLCMK 2 adherent epithelial cells. Thechemicalcompositionoftheessentialoil from P. ovatum leaves had a mixture of mono and sesquiterpenes and its major constituents were spathulenol, caryophyllene oxide, β-caryophyllene and myrcene. Since diseases caused by the parasites under investigation affect millions of people worldwide, results of this study are relevant because thechemicalcompositionoftheessentialoil from P. ovatum leaves was described for the first time. It is noteworthy that, in the face of medical advances, plants are still considered promising sources and bioactive compounds found in essential oils from plants ofthe genus Protium may serve as prototypes for the development of new antiparasitic drugs.
CONTENT ANDCOMPOSITIONOFTHEESSENTIALOILOF FIVE ACCESSES OF MENTRASTO. This study aimed to analyse the content andthecompositionoftheessentialoilof five accesses of mentrasto (Ageratum conyzoides). Five accesses of mentrasto with three repetitions were used. The accesses were obtained in Mariana-MG (AMA), Piranga-MG (API), Visconde do Rio Branco (ARB) and Viçosa-MG (AVB and AVP). Theessentialoil was obtained by hydrodistillation andthe identification oftheoil components by CG and GC/MS. The ARB access presented the higher essentialoil content, that is 0.70% (P < 0.05). Eleven chemical compounds were identified. The precocene I was the main constituent in the access API, andthe precocene II was the main constituent in the accesses AMA, ARB, AVB and AVP.
GC analysis oftheessentialoil was carried out on Agilent 5975 GC-MSD system and Agilent 6890N GC system with flame ionization detector (FID) and a HP-Innowax FSC column (60 m x 0.25 mm, 0.25 m film thickness). Helium was used as a carrier gas at 0.8 mL/min. The injector and detector temperatures were 250°C and 300°C, respectively. GC oven temperature was at 60°C for 10 min, then programmed to 220°C at a rate of 4°C/min and kept in this temperature for 10 min and finally programmed to 240°C at a rate of 1°C/min with a final hold time of 80 min. The split ratio was 40:1. Mass spectrums were taken at 70 eV and mass range was from m/z 35 to 450. In order to obtain same elution order with GC-MS, simultaneous injection was done by using the same column and appropriate operational conditions.
total oil from leaves. The identified compounds are listed in Table 1, according to their elution order on a VF-5 capillary column. Theoil contains a complex mixture consisting of mainly oxygenated mono- and sesquiterpenes, and mono- and sesquiterpene hydrocarbons. The major compounds detected in the leaves oil were phytol (27.25%), a- humulene (14.21%), spathulenol (12.12%), 1- octen-3-ol (8.21%), eugenol (6.69%), phenylethyl alcohol (5.81%) and caryophyllene oxide (2.6%), as shown in Table 1. Several papers reported that all these compounds possess significant antioxidant activity in several model systems [ 16,17 ]. It is also
T h e a n t i o x i d a n t a c t i v i t y t h r o u g h t h e phosphomolybdenum complex reduction method was performed using the standard solutions of ascorbic acid and rutin, that were prepared at the concentration of 200 μg / mL in methanol and 0.5% DMSO, as well as the samples (Prieto, Pineda, Aguilar, 1999). Aliquots of 0.3 mL were added to 3 mL ofthe phosphomolybdenum reagent (0.1 M tribasic sodium phosphate (28 mL), 0.03 M ammonium tetrahydrate molybdate solution (12 mL), 3 M sulfuric acid (20 mL) and water until complete 100 mL). The tubes were closed and brought to the thermostated bath at 95 ºC for 90 min. After cooling, the absorbances were obtained in 96-well microplates with round bottoms by reading in the Multiskan FC spectrophotometer, Thermo Scientific® at wavelength 695 nm. The antioxidant capacity ofthe samples was expressed in relative antioxidant activity (AAR%), in relation to the standards, using the equation: AAR% = [(Sample Absorbance - Absorbance of White)/ (Absorbance of Standard - Absorbance of White)] X 100 The variance ofthe obtained results was evaluated by the ANOVA test andthe difference between the means verified by the test (t) by Scott and Knott (p <0.05).