Molecular and chemical characterization of vetiver, Chrysopogon zizanioides (L.) Roberty, germplasm.

Due to the economic interests in vetiver, Chrysopogon zizanioides (L.) Roberty, molecular and chemical studies are essential to generate information for its sustainable exploitation. The aim of this study was to undertake a molecular and chemical characterization of vetiver accessions of the active germplasm bank of the Universidade Federal de Sergipe. The molecular characteristics of the accessions were studied using amplified fragment length polymorphism markers, with a total of 14 primer combinations that generated 442 loci, allowing us to observe that these accessions have similar genomes. The vetiver accessions were divided into three distinct groups, where accession UFS-VET005 was the most differentiated and accession UFS-VET004 had the lowest essential oil content (0.70%). The content of the chemical constituents of the essential oils was observed to vary, with a predominance of khusimol, which ranged from 18.97 to 25.02%. It was possible to divide the vetiver accessions into two groups based on chemical composition, and these groups do not correlate with the molecular grouping. Therefore, it is necessary to perform molecular and chemical analyses to characterize vetiver accessions.

Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts.

This study aimed to evaluate the chemical composition of the essential oils from three genotypes of Lippia gracilis Schauer (Verbenaceae) and investigate the cytotoxic activities of these oils. Essential oils were extracted from the leaves using a Clevenger-type apparatus, and chemical analysis was performed using a gas chromatograph coupled to a mass spectrometer and flame ionization detector. 3T3, MRC5, B16, HeLa, and MCF-7 cell lines were used to study the in vitro cytotoxicity of the essential oils, and the level of cell death was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test with three replicates. The cytotoxic activity was expressed as the concentration that inhibited 50% of cell growth. The main compound in the essential oil of LGRA-106 was thymol (40.52%), while LGRA-109 and LGRA-201 contained 45.84 and 32.60% carvacrol, respectively, as their major compound. The essential oils of L. gracilis showed cytotoxic activity against both normal and tumor cells at concentrations below 100 µg/mL; this demonstrated the antitumor potential of these essential oils, which should be further investigated.

Effects of essential oils on Aedes aegypti larvae: alternatives to environmentally safe insecticides.

The essential oils from leaves of Hyptis fruticosa (Lamiaceae) Salzm., H. pectinata (Lamiaceae) Poit., and Lippia gracilis (Verbenaceae) HBK were investigated for their larvicidal activity against Aedes aegypti and analyzed by GC/MS. Fifty-nine compounds, representing 91.28-98.39% of the essential oils, have been identified. A standard solution was used to make 20 mL solutions ranging from 30 to 2000 ppm. Twenty larvae between third and fourth stages were added to the essential oil solution. A mortality count was conducted 24 h after treatment. Essential oils LC50 and their confidence limits at 95% probability were calculated by the methods of Reed-Muench and Pizzi, respectively. The essential oil of Lippia gracilis showed potent insecticidal effect against Aedes aegypti larvae, the vector of dengue fever. Carvacrol and caryophyllene oxide were the main responsible for the activity of L. gracilis and H. pectinata. Minor compounds are probably acting synergistically to achieve H. fruticosa activity.

Prevalence and screening of active tuberculosis in a prison in the South of Brazil.

SETTING: Tuberculosis (TB) remains a challenge in Brazil, particularly among prison inmates. OBJECTIVE: To assess TB prevalence by active case finding in a public prison in southern Brazil. DESIGN: Prison inmates were screened for TB using the presence of cough and chest X-ray (CXR) from October 2014 to August 2016. Presence of cough, irrespective of duration, and abnormal CXRs were further investigated using laboratory tests. RESULTS: Of 10 326 inmates screened, 196 had confirmed TB (1898/100 000 inmates screened). At the first screening, 1759 inmates presented with cough only, 16 of whom had TB; among those with only abnormal CXR (n = 1273), 92 had TB. Xpert was positive in 155 patients, and negative in 15; these results were confirmed using culture. The remaining 26 patients did not undergo Xpert testing and were confirmed using microscopy (27%), culture (42%) or both (31%). CONCLUSION: The combined use of symptom screening (cough) and CXR was much more effective in maximising TB yield than using either method alone. If patients presenting with cough alone had not been investigated, 10% of TB patients would have been missed; if those with abnormal CXR but no cough had not been investigated, 51% of TB patients would have been missed. We detected high TB prevalence in this prison by using active case finding.

Cardiovascular effects of Hyptis fruticosa essential oil in rats.

In non-anesthetized normotensive rats, Hyptis fruticosa essential oil (HFEO, 5, 10, 20 and 40 mg/kg; i.v.) induced hypotension associated with tachycardia. In intact and isolated rings of rat superior mesenteric artery (control), HFEO (1-1000 microg/ml, n=6, cumulatively) induced concentration-dependent relaxations of tonus induced by 10 microM phenylephrine (Phe) (pD(2)=2.6+/-0.27; E(max)=64+/-8.3%). In denuded endothelium pre-contracted rings with Phe or K(+)-depolarizing solution (80 mM), the concentration-response curves to HFEO were not shifted (pD(2)=2.3+/-0.25 and 2.3+/-0.28, respectively), but their maximal responses were significantly (P<0.05 vs control) increased (E(max)=122.3+/-18.2% and 92+/-3.6%, respectively). HFEO was also capable of antagonizing the concentration-response curves to CaCl(2) (3 microM-30 mM) in a dose-dependent manner.

Central nervous system effects of the crude extract of Erythrina velutina on rodents.

The aqueous extract (AE) of Erythrina velutina prolonged the sleep duration induced by sodium pentobarbital (control: 6.4 +/- 1.2 min; extract 10 mg/kg, 47.1 +/- 3.9 min; extract 100 mg/kg, 109.4 +/- 7.2 min; F = 243, P < 0.001). In the open field, the extract at the doses of 10 and 50 mg/kg did not changed the number of crossings, rearings nor groomings. On the other hand, at the dose of 200 mg/kg it reduced the number of crossings (q = 6.25, P < 0.05) and groomings (q = 3.91, P < 0.05). When exposed during three consecutive days to the open field, the control animals showed habituation for crossings (F = 17.56, P < 0.001) and rearings (F = 14.01, P < 0.001). The same was not true for animals treated with 10 mg/kg of the extract (crossings: F = 3.59, P > 0.05; rearings: F = 3.62, P > 0.05). At the same dose, the extract blocked the acquisition of foot shock memory (P = 0.9219) when compared to the control values (P = 0.0078). Our data showed that the crude extract of Erythrina velutina at lower doses interferes with mnemonic process for different tasks, while at higher doses, the sedative and neuromuscular blocking actions are the main effects.

Influence of storage time and temperature on the chemical composition of the essential oil of Hyptis pectinata L. Poit / Influência do tempo de armazenamento e da temperatura na composição química do óleo essencial de Hyptis pectinata L. Poit

ABSTRACT Hyptis pectinata, popularly known as “sambacaitá”, is a native medicinal herb used by local people for treating disease. Its pharmacological, antimicrobial, and medicinal properties have been reported in several studies. The essential oil of the leaves is rich in terpene compounds responsible for its properties. Because there are factors that can influence the chemical composition of essential oils, the objective of this study was to evaluate the influence of storage on Hyptis pectinata essential oil for 360 days at two different temperatures. The essential oil was distilled from dried leaves of seven-month-old plants using the hydrodistillation method, and the chemical composition was determined using GC-MS and GC/FID. The effects of storage on the essential oil Hyptis pectinata were tested over the course of a year (0, 15, 30, 60, 90, 120, 150, 180, 240, 300, and 360 days), using two temperatures: room (±32ºC) and freezer (-20ºC). The predominant compounds in the essential oil are β–caryophyllene, caryophyllene oxide, and germancrene D. The concentration of the chemical constituents of the essential oil varied depending on the storage temperature, and over the 360 days. The storage of the essential oil at room temperature over one year resulted in higher concentrations of β-elemene, α-copaene, germacrene D, caryophyllene oxide, and (E,E)-α-farnesene and lower content of α-humulene and β-caryophyllene, compared to the results obtained from storing the essential oil in a freezer. These results indicate that the essential oil should be stored in a freezer, which provides greater stability to the concentration of the chemical constituents.

RESUMO Hyptis pectinata, conhecida popularmente como Sambacaitá é uma erva medicinal nativa utilizada pela população local para o tratamento de enfermidades, cujas propriedades farmacológicas, antimicrobianas e medicinais foram relatadas em diversos estudos. O óleo essencial das folhas é rico em compostos terpênicos responsáveis pelas suas propriedades. Devido à existência de fatores que podem influenciar a composição química dos óleos essenciais, o objetivo deste trabalho foi avaliar a influência do armazenamento do óleo essencial Hyptis pectinata durante 360 dias em duas temperaturas diferentes. O óleo essencial foi destilado a partir de folhas secas de plantas de sete meses de idade, utilizando o método de hidrodestilação e a composição química foi determinada usando CG-EM e CG-DIC. Os efeitos do armazenamento no óleo essencial Hyptis pectinata foram testados ao longo de um ano (0, 15, 30, 60, 90, 120, 150, 180, 240, 300 e 360 dias), utilizando duas temperaturas: ambiente (±32ºC) e freezer (-20ºC). Os compostos predominantes no óleo essencial foram o β-cariofileno, óxido de cariofileno e germancreno D e houve variação na concentração dos constituintes químicos do óleo essencial, dependendo da temperatura de armazenamento, bem como ao longo dos 360 dias. O armazenamento do óleo essencial, à temperatura ambiente ao longo de um ano resultou em maiores concentrações de β-elemeno, α-copaeno, germacreno D, óxido de cariofileno e (E, E) -α-farneseno e menores teores de α-humuleno e β-cariofileno, quando comparado com os resultados obtidos a partir de armazenamento do óleo essencial no freezer. Estes resultados indicam que o óleo essencial deve ser estocado no freezer, o que propicia maior estabilidade na concentração dos constituintes químicos.

Composition and acaricidal activity of Lippia sidoides essential oil against two-spotted spider mite (Tetranychus urticae Koch).

The essential oils from accessions of Lippia sidoides Cham. (Verbenaceae) were characterized by GC and GC/MS and investigated for their acaricidal activity against the two-spotted spider mite (Tetranychus urticae Koch). Twenty-nine compounds were identified with potential acaricidal activity. Glass receptacles were used as test chambers. For each dose and exposure time combination, three replicates were used. Each replicate consisted of 30 adult females of T. urticae, 10 mites in each leaf disk of Canavalia ensiformis placed in a Petri dish. Increasing amounts of oil or terpene were applied on a blotting paper strip, fixed on the inner surface of the glass recipient cover, corresponding to 2, 4, 6, 8, and 10 microL/L of air, respectively. Exposure periods were 24, 48, and 72 h. Data obtained in these experiments were submitted to probit analysis. The essential oil of L. sidoides, thymol and carvacrol exhibited potent acaricidal activity against T. urticae.

Evaluation of the analgesic and anti-inflammatory effects of the essential oil of Lippia gracilis leaves.

AIM OF THE STUDY: The aim of the present study is to investigate the antinociceptive, anti-inflammatory, and antioxidant activities of essential oil (EO) of Lippia gracilis Schauer (Verbenaceae) leaves to support the medicinal uses claimed by folklore practitioners in the caatinga region (semi-arid) of Northeastern Brazil. MATERIALS AND METHODS: The chemical composition and antinociceptive and anti-inflammatory activities of the EO of Lippia gracilis leaves (50-200 mg/kg) were investigated. Antinociceptive activity of the EO was evaluated by writhing test. Anti-inflammatory activity of the EO was evaluated using paw oedema and peritonitis methods. RESULTS: Oral treatment with the EO of Lippia gracilis leaves elicited inhibitory activity on acetic acid effect at 50, 100, and 200 mg/kg (30.33+/-2.36, 25.20+/-1.48, and 21.00+/-1.54 abdominal writhes, respectively, P<0.05), as compared with the control group (36.73+/-1.92 writhes). The compound acetylsalicylic acid (ASA, 300 mg/kg) inhibited the acetic acid-induced writhing (12.67+/-0.50 abdominal writhes, P<0.001). Carrageenan-induced oedema formation was reduced with the EO of Lippia gracilis leaves at 200 mg/kg (0.72+/-0.06 mL h, P<0.001) and by the reference compound ASA (300 mg/kg, 0.85+/-0.04 mL h, P<0.001), as compared with the control group (1.76+/-0.06 mL h). Leukocyte migration into the peritoneal cavity induced by carrageenan was reduced with the EO of Lippia gracilis leaves at 50, 100, and 200 mg/kg (13.81+/-0.61, 11.77+/-0.91, and 10.30+/-0.60 leukocytes x 10(6)/mL, respectively, P<0.01), and by the compound dexamethasone (2 mg/kg, 5.34+/-0.33 leukocytes x 10(6)/mL, P<0.001), as compared with the control group (16.71+/-0.54 leukocytes x 10(6)/mL). The analyses of the essential oil allowed the identification of Lippia gracilis as a thymol-p-cymene chemotype (32.68% and 17.82%, respectively). CONCLUSIONS: The EO of Lippia gracilis leaves shows antinociceptive and anti-inflammatory activities.

Antinociceptive activity of the volatile oils of Hyptis pectinata L. Poit. (Lamiaceae) genotypes.

Hyptis pectinata L. Poit (Lamiaceae) is known popularly in Brazil as "sambacaita" or "canudinho" and is used in the treatment of inflammations, bacterial infections and ache. The antinociceptive activity of the volatile oils of six genotypes, at doses of 100, 200 and 400mg/kg body wt., were investigated using abdominal writhe models induced by acetic acid and hot plate tests. The volatile oils of all the genotypes are composed mainly of sesquiterpenoids. All the genotypes showed antinociceptive effects in both models used; the SAM002 genotype showed the major inhibitory effect at dose of 100mg/kg body wt. These results suggest that the volatile oil of H. pectinata has peripheral (writhe reduction) and central (time delay of thermal reaction) effects. These observations indicate that H. pectinata may be useful as an analgesic drug.

Phythochemical screening and anticonvulsant activity of Cymbopogon winterianus Jowitt (Poaceae) leaf essential oil in rodents.

Cymbopogon winterianus (Poaceae) is used for its analgesic, anxiolytic and anticonvulsant properties in Brazilian folk medicine. This report aimed to perform phythochemical screening and to investigate the possible anticonvulsant effects of the essential oil (EO) from fresh leaves of C. winterianus in different models of epilepsy. The phytochemical analysis of EO showed presence of geraniol (40.06%), citronellal (27.44%) and citronellol (10.45%) as the main compounds. A behavioral screening demonstrated that EO (100, 200 and 400mg/kg; ip) caused depressant activity on CNS. When administered concurrently, EO (200 and 400mg/kg, ip) significantly reduced the number of animals that exhibited PTZ- and PIC-induced seizures in 50% of the experimental animals (p<0.05). Additionally, EO (100, 200 and 400mg/kg, ip) significantly increased (p<0.05) the latencies of clonic seizures induced by STR. Our results demonstrated a possible activity anticonvulsant of the EO.

Adubação orgânica na produção de biomassa de plantas, teor e qualidade de óleo essencial de orégano (Origanum vulgare L. ) em cultivo protegido / Organic fertilization on phytomass production and essential oil content and quality of oregano (Origanum vulgare L. ) under protected cultivation

Origanum vulgare L., conhecida como orégano, é considerada planta condimentar largamente usada na culinária. No Brasil poucas são as pesquisas com esta espécie visando maximização das técnicas de cultivo. Objetivou-se avaliar os efeitos de diferentes doses dos adubos orgânicos bovino e aves no crescimento de plantas, teor de clorofila, teor, rendimento e qualidade do óleo essencial de orégano cultivado sob estufa. Foram conduzidos dois ensaios, sendo um com esterco bovino e o outro com esterco de aves (Poedeira). Plantas de orégano (Origanum vulgare L.) foram cultivadas em vasos de 10 L submetidas aos seguintes tratamentos: Ensaio I: Esterco bovino: 1) Solo sem adubação (controle); 2) solo + 3,0 kg m-2 de esterco bovino; 3) solo + 6,0 kg m-2 de esterco bovino; 4) solo + 9,0 kg m-2 de esterco bovino; 5) solo + 12,0 kg m-2 de esterco bovino; Ensaio II - Esterco de aves: 1) solo sem adubação (Controle); 2) solo + 1,5 kg m-2 de esterco de galinha; 3) solo + 3,0 kg m-2 de esterco de galinha; 4) solo + 4,5 kg m-2 de esterco de galinha e 5) solo + 6,0 kg m-2 de esterco de galinha. Ambos os ensaios foram conduzidos em delineamento blocos ao acaso com 4 repetições e a parcela experimental composta de 4 vasos. Foi observado que as doses de adubos bovino e avícola influenciaram significativamente no crescimento das plantas, rendimento e composição química do óleo essencial de orégano.

Origanum vulgare L., known as oregano or wild marjoram, is an aromatic plant widely used in cookery. In Brazil, there are few studies with this species aimed at improving the cultivation techniques. The present study was carried out to evaluate the effect of different levels of cattle and chicken organic manure on plant growth, chlorophyll content, besides essential oil content, yield and quality of oregano grown in a greenhouse. Two assays were carried out, one of them used cattle manure and the other, chicken manure (Hen). Oregano (Origanum vulgare L.) plants were grown in 10L-pots and subjected to the following treatments: Assay I: Cattle manure: 1) Soil without fertilization (control); 2) Soil + 3.0 kg m-2 cattle manure; 3) Soil + 6.0 kg m-2 cattle manure; 4) Soil + 9.0 kg m-2 cattle manure; and 5) Soil + 12.0 kg m-2 cattle manure; Assay II: Chicken manure: 1) Soil without fertilization (control); 2) Soil + 1.5 kg m-2 chicken manure; 3) Soil + 3.0 kg m-2 chicken manure; 4) Soil + 4.5 kg m-2 chicken manure; and 5) Soil + 6.0 kg m-2 chicken manure. The experimental design for both assays was in randomized blocks with four replicates and four pots per plot. Cattle and chicken manure levels significantly influenced oregano plant growth besides essential oil yield and chemical composition.