Essential oils biological activity of the shrub Lippia alba (Verbenaceae)

Introduction: Lippia alba is an aromatic species belonging to the Verbenaceae family. Its essential oils have been used in different industries, because of its biological properties. Objective: Identify the perspectives of the biotechnological applications of Lippia alba essential oils. Methodology: A scoping review was conducted on the biological activity of Lippia alba essential oils registered until October, 2018 in EBSCO, Embase, Pubmed, Scopus, SciELO, and Lilacs databases. Results: Chemotypes I and III have been reported for different biological activities from the evaluations performed on microorganisms, fish, arthropods, small mammals, and cell lines; fundamentally associated with antibacterial, antifungal, cytotoxic, antioxidant, and sedative effects, among others. Records focused mainly on the health, fishing industry, and agrifood sectors. Conclusion: Studies on the effect of essential oil are promising, but do not reflect a continuity of the research toward prototypes or finished commercial products. Research groups must unify evaluation methodologies and include in all studies the relationship between phytochemical and biological activity for the meta-analyses to be possible. Likewise, they must join efforts through the National System for Agricultural Innovation (SNIA, for the term in Spanish) to generate finished products that impact upon society and facilitate progress in the country's bio-economy.

Introducción: Lippia alba es una especie aromática perteneciente a la familia Verbenaceae, cuyos aceites esenciales han sido empleados en diferentes industrias dada sus propiedades biológicas. Objetivo: Identificar las perspectivas de aplicación biotecnológica de los aceites esenciales de Lippia alba. Métodos: Se realizó una revisión sistemática exploratoria de la literatura sobre la actividad biológica de aceites esenciales de Lippia alba registrada hasta octubre 2018 en las bases de datos EBSCO, Embase, Pubmed, Scopus, SciELO, y Lilacs. Resultados: Los quimiotipos I y III han sido reportados para diferentes actividades biológicas a partir de evaluaciones realizadas en microorganismos, peces, artrópodos, pequeños mamíferos, y líneas celulares; fundamentalmente asociadas con efectos antibacterial, antifúngico, citotóxico, antioxidante y sedante, entre otros. Los registros se enfocaron principalmente a los sectores salud, industria pesquera y agroalimentaria. Conclusión: Los estudios sobre el efecto del aceite esencial son promisorios, pero no reflejan una continuidad de la investigación hacia prototipos o productos comerciales acabados. Los grupos de investigación deben unificar metodologías de evaluación e incluir en todos los estudios la relación entre fitoquímica-actividad biológica, para que los metaanálisis sean posibles. De igual manera, deben aunar esfuerzos por medio del Sistema Nacional de Innovación Agropecuaria (SNIA), para generar productos acabados que impacten en la sociedad y faciliten el avance de la bioeconomía del país.

Fungicidal potential of essential oils in control of Fusarium spp. and Sclerotinia sclerotiorum / Potencial fungicida de óleos essenciais no controle de Fusarium spp. e Sclerotinia sclerotiorum

The use of highly toxic pesticides to control soil pathogens, such as Fusarium spp. and Sclerotinia sclerotiorum has generated concern, due to the irreversible impacts caused on the environment, in addition to selecting resistant isolates. In this way, essential oils appear as an efficient alternative in control of diseases. Facing the problem of soil pathogens control and high antimicrobial fungicide that essential oils present, this work aimed to evaluate the in vitro fungicidal potential of essential oils in control of Fusarium spp. and S. sclerotiorum. A completely randomized design, factorial scheme 2×4×8 was used, with two isolates (Fusarium spp. and S. sclerotiorum), four essential oils (Aloysia citriodora, Cymbopogon winterianus, Lippia alba and Ocimum americanum), eight essential oil concentrations (0.0; 0.2; 0.4; 0.6; 0.8; 1.0; 1.2 and 1.4 ?L·mL-1), and ten replicates. The essential oils inhibited mycelial growth of the fungi in different concentrations, being their potential justified by the presence of antifungal chemical compounds. Essential oils of A. citriodora, C. winterianus, L. alba and O. americanum present high fungicidal potential, being viable alternatives for formulation of commercial products, boosting the pesticides industry.(AU)

O uso de pesticidas com alta toxicidade para controlar patógenos do solo, como Fusarium spp. e Sclerotinia sclerotiorum, tem gerado preocupação, devido aos impactos irreversíveis causados no meio ambiente, além de selecionar isolados resistentes. Dessa forma, os óleos essenciais surgem como uma alternativa eficiente no controle de doenças. Diante da problemática de controle de patógenos do solo e alto potencial antimicrobiano que os óleos essenciais possuem, este trabalho teve como objetivo avaliar o potencial fungicida de óleos essenciais no controle de Fusarium spp. e S. sclerotiorum, in vitro. Utilizou-se um delineamento inteiramente randomizado, esquema fatorial 2×4×8, com dois isolados (Fusarium spp. e S. sclerotiorum), quatro óleos essenciais (Aloysia citriodora, Cymbopogon winterianus, Lippia alba e Ocimum americanum) e oito concentrações de óleo essencial (0,0; 0,2; 0,4; 0,6; 0,8; 1,0; 1,2 e 1,4 ?L·mL-1), com dez repetições. Os óleos essenciais inibiram o crescimento micelial dos fungos em diferentes concentrações, sendo seu potencial justificado pela presença de compostos químicos antifúngicos. Os óleos essenciais de A. citriodora, C. winterianus, L. alba e O. americanum apresentam alto potencial fungicida, sendo alternativas viáveis para formulação de produtos comerciais, impulsionando a indústria de agrotóxicos.(AU)

Citral chemotype of the Lippia alba essential oil as an additive in simulated transport with different loading densities of tambaqui juveniles / Óleo essencial de Lippia alba quimiotipo citral como aditivo em transporte simulado de juvenis de Tambaqui com diferentes densidades

ABSTRACT: The objective of this study was to evaluate the efficacy of the citral chemotype of Lippia alba essential oil (EOLA) as an additive in a simulated transport with different loading densities of tambaqui (Colossoma macropomum) juveniles. Simulated transport of tambaqui juveniles at three loading densities (30, 60 and 90 fish/L) was carried out either with only water (WC) or 20 μL.L-1 EOLA. Dissolved oxygen levels, pH, conductivity and temperature of the water were measured at the beginning and end of the simulated transport. Treatment with EOLA did not change survival or dissolved oxygen levels at the end of simulation. The pH of the water at the end of simulation with EOLA was lower than in that of WC but did not change with loading density. Water conductivity increased with loading density; however, it was lower in the groups with EOLA. The addition of EOLA can be useful in long-term transportation at high loading densities, but further studies related to tambaqui physiology are required.

RESUMO: O objetivo deste estudo foi avaliar a eficácia do óleo essencial de Lippia alba quimiotipo citral (EOLA) como aditivo em transporte simulado com diferentes densidades de carga de juvenis de tambaqui (Colossoma macropomum). O transporte simulado de juvenis de tambaqui em três densidades de carga (30, 60 e 90 peixes / L) foi realizado apenas com água (WC) ou 20 μL.L-1 EOLA. Níveis de oxigênio dissolvido, pH, condutividade e temperatura da água foram medidos no início e no final do transporte simulado. O tratamento com EOLA não alterou a sobrevivência ou os níveis de oxigênio dissolvido no final da simulação. O pH da água no final da simulação com o EOLA foi menor que o do WC, mas não mudou com a densidade de carga. A condutividade da água aumentou com a densidade de carga, no entanto foi menor nos grupos com EOLA. A adição de EOLA pode ser útil no transporte de longo prazo em altas densidades de carga, mas são necessários mais estudos relacionados à fisiologia do tambaqui.

Adulterantes presentes en plantas medicinales de mercados municipales en Costa Rica / Presence of adulterants in medicinal plants of municipal markets in Costa Rica

RESUMEN La carencia de una regulación sobre las plantas medicinales y el escaso control de calidad dado en Costa Rica ponen en riesgo la salud de la población. El objetivo del presente trabajo fue determinar la presencia de adulterantes en manzanilla (Matricaria chamomilla) y juanilama (Lippia alba) de venta a granel en mercados de San José, Heredia y Cartago, Costa Rica. Se recolectaron muestras de manzanilla y juanilama en diferentes puntos de venta dentro de los mercados de Cartago, Heredia y San José. Las muestras se analizaron en fresco y a través de los criterios establecidos por la OMS, para el control de calidad de productos herbarios, se estableció la cuantificación de materiales ajenos al producto comerciado como: insectos, mezclas de minerales del suelo, plástico, vidrio y moho. Los resultados fueron analizados con Excel 2010 y el programa estadístico IBM SPSS 22. En mercados de Heredia y San José se venden productos que no corresponden a la especie terapéutica ofrecida. La manzanilla es vendida principalmente en condiciones frescas, mientras que la juanilama varía su condición dependiendo del mercado. Hubo presencia de adulterantes, como insectos, tierra, plásticos y mohos, en las plantas de manzanilla y juanilama de venta en las provincias de Cartago, Heredia y San José.

SUMMARY The lack of regulation and quality control in Costa Rica on the medicinal plants has risks in public health. The aim of these work was to determinate presence of contaminants in chamomile (Matricaria chamomilla) and juanilama (Lippia alba) commercialized in the municipal markets in San José, Heredia and Cartago, Costa Rica. They were collected samples of chamomile and juanilama in different sale points within the Cartago, Heredia and San José markets. The samples were analyzed fresh and through the criteria established by WHO; for the quality control of herbal products, the quantification of materials foreign to the traded product was established: insects, mixtures of soil minerals, plastic, glass and mold. The results were analyzed with the statistical program SPSS 22. In some markets of Heredia and San José products do not correspond to the therapeutic species offered, and they were sold. Chamomile is sold mainly in fresh conditions, while the juanilama varies its condition depending on the market. There was presence of adulterants, such as insects, soil, plastics and molds, in plants chamomile and juanilama sold in the provinces of Cartago, Heredia and San José.

Effect of Lippia alba essential oil administration on obesity and T2DM markers in Wistar rats / Efecto de la administración del aceite esencial de Lippia alba sobre la obesidad y los marcadores T2DM en ratas wistar

SUMMARY Introduction: Lippia alba (Mill) N.E. Brown (Verbenaceae) is an aromatic plant from Central America, South America, and the Caribbean, it is traditionally used by the Colombian population to treat various diseases such as diabetes and hypertension. The purpose of this research was to evaluate the metabolic effects of Lippia alba essential oil (EO) oral administration on obesity and diabetes markers in Wistar rats. Methods: control and Streptozotocin (STZ) diabetes induced rats were used to evaluate the EO metabolic effects. Glucose and triglycerides were measured using commercial colorimetric kits, the animals' weight was followed for 21 days treatment and TNF-α and adiponectin concentration was determined with ELISA technique. Results: The consumption of EO shows body weight gain regulation, lower glucose and cholesterol levels in normal rats and lower TNF- α in comparison with the Glibenclamide treated rats between the STZ diabetic groups. No toxic effects were founded. Conclusions: The EO exerts a benefical metabolic effect in rats, therefore it is interesting to be evaluate a future in human beings with T2DM or overweight.

RESUMEN Introducción: Lippia alba (Mill) N.E. Brown (Verbenaceae) más conocida como pronto alivio, es una planta aromática de Centro, Sur América y el Caribe que se utiliza tradicionalmente en Colombia para el tratamiento de diversas enfermedades, como la diabetes e hipertensión. El objetivo del trabajo fue evaluar el efecto metabólico del aceite esencial de Lippia alba (Mill), administrado oralmente, sobre moléculas relacionadas con obesidad y diabetes en ratas Wistar. Métodos: Se pesaron los animales diariamente. Después de 21 días de tratamiento con el AE se determinó en plasma la glucosa, triglicéridos con kits comerciales y las adipocitoquinas (adiponectina y factor de necrosis tumoral alfa (TNFα) marcadores de resistencia a la acción de la insulina) por la técnica de ELISA. Resultados: El consumo de AE mostró una regulación en la ganancia de peso corporal y disminución en los niveles de glucosa y triglicéridos en los animales normales que recibieron el AE. Dentro de los grupos con diabetes inducida, el grupo tratado con AE mostró menores valores de TNF-α comparado con el grupo tratado con glibenclamida. Conclusiones: El AE ejerce un efecto benéfico en el metabolismo de los animales, por lo tanto, es interesante para ser evaluado en seres humanos con diabetes o sobrepeso.

Pharmacological effects of different chemotypes of Lippia alba (Mill.) N.E. Brown / Efectos farmacológicos de diferentes quimiotipos de Lippia alba (Mill.) N.E. Brown

The Lippia alba species consists of an aromatic plant used in Brazilian traditional medical practice and in the medical practice of several countries as well. Presenting a wide variability in its essential oil chemical composition, the Lippia alba is classified in chemotypes, or chemical races, according to the major constituents contained in its essential oil. Considering the quali and quantitative distribution of the components in the essential oil affect directly its pharmacological properties, which are presented in the medicinal species, this paper proposes a scientific literature review to correlate both biological and pharmacological properties presented by L. alba according to its chemical constitution.

Lippia alba es una planta aromática utilizada en la medicina tradicional de Brasil y de varios países. Con una gran variabilidad en la composición química de su aceite esencial, se clasifica en quimiotipos, o razas químicas, de acuerdo con los constituyentes mayoritarios presentes en el aceite esencial. Dado que la distribución cualitativa y cuantitativa de los componentes del aceite esencial afecta directamente a las propiedades farmacológicas presentadas por la especie medicinal, este trabajo propone realizar una revisión en la literatura científica para correlacionar las propiedades biológicas y farmacológicas de los quimiotipos presentes en el aceite essencial de la L. alba.

Antimicrobial activity of essential oils from Lippia alba, Lippia sidoides, Cymbopogon citrates, Plectranthus amboinicus, and Cinnamomum zeylanicum against Mycobacterium tuberculosis / Atividade antimicrobiana de leos essenciais de Lippia alba, Lippia sidoides, Cymbopogon citrates, Plectranthus amboinicus, e Cinnamomum zeylanicum em Mycobacterium tuberculosis

ABSTRACT: The rise in cases of antibiotic-resistant Mycobacterium tuberculosis has become a major obstacle to the effective control of tuberculosis (TB) worldwide. Essential oils (EO) are complex mixtures that may contain between 20 and 60 components, with two or three major compounds at relatively high concentrations (20-70%) that are responsible for their pharmacological properties. The objective of this study was to assess the antimicrobial activity of the EOs, bushy lippia (Lippia alba), rosemary pepper (Lippia sidoides), lemon grass (Cymbopogon citratus), Mexican mint or Indian borage (Plectranthus amboinicus), and true cinnamon (Cinnamomum zeylanicum), against Mycobacterium tuberculosis H37Rv. Chemical characterization of the EOs was performed by gas chromatography coupled to mass spectrometry. The minimum inhibitory concentration (MIC) was determined by the microdilution-based resazurin microtiter assay. Four EOs were able to inhibit the growth of M. tuberculosis, with MICs of 286.5±130.2μg/mL (C. zeylanicum), 299.5±117.2μg/mL (L. sidoides), 351.6±39.06μg/mL (P. amboinicus), and 1,250μg/mL (C. citratus). Only the EO of L. alba showed no antimycobacterial activity at the tested concentrations, with an MIC greater than 1,250µg/mL. Results of this study suggested that C. zeylanicum, L. sidoides, and P. amboinicus could be important sources of bactericidal compounds against M. tuberculosis and require further investigation. The activity against M. tuberculosis of these three EOs has not been reported previously. The results show the high potential of the tested antimycobacterial EOs, making them a promising alternative for TB treatment. This data also confirms the importance of bioprospecting studies for active substances with antimycobacterial activity, which are still scarce.

RESUMO: O aumento no número de casos de Mycobacterium tuberculosis resistentes tem se tornado um grande obstáculo no controle efetivo da tuberculose (TB) mundialmente. Os óleos essenciais (OE), que são misturas complexas que podem conter entre 20 a 60 componentes, apresentam dois ou três compostos principais, em concentrações relativamente elevadas, 20 a 70%, que são responsáveis pelas suas propriedades farmacológicas. O objetivo deste estudo foi avaliar a atividade antimicobacteriana dos seguintes óleos essenciais (OEs) em Mycobacterium tuberculosis H37Rv: erva-cidreira (Lippia alba), alecrim-pimenta (Lippia sidoides), capim-limão (Cymbopogon citratus), orégano (Plectranthus amboinicus) e canela (Cinnamomum zeylanicum). A caracterização química dos OEs foi realizada por cromatografia gasosa acoplada a espectrometria de massa. A Concentração Inibitória Mínima (CIM) foi determinada pela técnica de microdiluição da resazurina. Quatro OEs foram capazes de inibir o crescimento de M. tuberculosis, com CIM de 286,5±130,2μg/mL (C. zeylanicum), 299,5±117,2μg/mL (L. sidoides), 351,6±39,06μg/mL (P. amboinicus) e 1250μg/mL (C. citratus). Somente o OE de L. alba não mostrou atividade antimicobacteriana nas conscentrações testadas, considerando CIM maiores que 1250µg/mL. Os resultados deste estudo sugerem que L. sidoides, C. zeylanicum e P. amboinicus podem ser fontes importantes de compostos bactericidas contra M. tuberculosis e prováveis candidatos a serem investigados. A atividade contra M. tuberculosis desses três OEs não foi relatada em estudos anteriores. Os resultados mostram o elevado potencial antimicobacteriano dos OEs analisados, fazendo deles uma alternativa promissora para o tratamento da TB. Os resultados obtidos demonstraram a importância de pesquisas para bioprospecção de substâncias ativas com ação antimicobacteriana, que ainda são escassas.

Caracterización química y evaluación de la actividad antifúngica del aceite esencial foliar de Lippia alba contra Colletotrichum gloeosporioides

El objetivo de este estudio fue caracterizar y evaluar in vitro la eficiencia del aceite esencial foliar de Lippia alba contra Colletotrichum gloeosporioides. Se extrajo el aceite esencial a partir de las hojas mediante el método de hidrodestilación asistida por microondas (MWHD) y se identificaron sus metabolitos mediante una cromatografía de gases acoplada a espectrometría de masas (GC/MS). La evaluación in vitro se realizó empleando 4 concentraciones del aceite esencial (500, 1000, 3000 y 10000 ppm) diluidas en acetona; se utilizó un control positivo con Benomil 1 g/L, un control negativo con acetona y un testigo absoluto; la eficiencia del aceite se midió mediante el porcentaje de índice antifúngico (%I.A). El componente mayoritario en el aceite esencial fue el citral (34.62 a 40.03%) y que el mayor %I.A se encontró a la concentración de 10000 ppm (97.8%), muy similar a la del Benomil (100%). Lo anterior demostraría la eficacia del aceite esencial de L. alba para controlar a C. gloeosporioides, y su posible uso como fungicida biológico.

The aim of this study was to characterize and evaluate the in vitro efficiency of leaf essential oil of Lippia alba against Colletotrichum gloeosporioides. Essential oil from the leaves was extracted by the method of assisted microwave hydrodistillation (MWHD) and metabolites were identified by gas chromatography coupled to mass spectrometry (GC/ MS). The in vitro evaluation was performed using 4 concentrations of essential oil (500, 1000, 3000 and 10000 ppm) diluted in acetone; positive control with Benomyl 1 g /L, a negative control with acetone and absolute control was used; oil efficiency was measured by the percentage of antifungal index (% IA). It was found that the major component was in the essential oil citral (34.62 to 40.03%) and the highest %I.A was found to 10000 ppm concentration (97.8%) very similar to Benomyl (100%). This would demonstrate the efficacy of the L. alba essential oil to control C. gloeosporioides, and its possible use as a biological fungicide.

Composición química y toxicidad aguda oral del aceite esencial de Lippia alba en ratones / Chemical composition and oral acute toxicity of Lippia alba essential oil in mice

La Lippia alba (L. alba), de la familia Verbenaceae, conocida popularmente como “Cidrón” o “poleo”; dependiendo de su ubicación geográfica, el suelo donde se desarrolla y tejido o órgano, presenta aceites esenciales en su composición, que son empleados en medicina natural y tradicional para el tratamiento de trastornos digestivos, respiratorios, enfermedades de la piel y sedante, a través de infusión o decocción de hojas, partes aéreas o flores. Por lo que posee un elevado potencial terapéutico, siendo importante detectar posibles efectos tóxicos post administración. Por lo cual, el objetivo del estudio fue evaluar la composición química y la toxicidad aguda oral del aceite esencial de L. alba a dosis única y dosis repetida. Se extrajo el aceite esencial de las hojas y partes aéreas, por destilación con arrastre de vapor utilizando una Trampa de Clevenger y se analizó por cromatografía de gases acoplada a detector de masas. Se evaluó la toxicidad, utilizando ratones NRMI de ambos sexos y las Guías OECD 423 y 407. Se obtuvieron 34 constituyentes del aceite esencial, los mayoritarios fueron: geranial (45,30%), neral (30,23%), beta-farnesol (7,78%), cariofileno (2,85%), azuleno (2,52%) y beta- irceno (2,38%). Se evidenciaron (Guía OECD 423) signos y síntomas de daño neurológico y motor moderado (900 mg/Kg del aceite esencial) con 100% de supervivencia y severo (1500 mg/ Kg) letales para los machos y, con 2000 mg/Kg todos los animales fallecieron. Se concluye que en el aceite esencial predominaron los constituyentes geranial y neral; clasificados como quimiotipo “citral” y que de acuerdo a la Clasificación de Toxicidad del Sistema Globalmente Armonizado de Clasificación y Etiquetado de Productos Químicos (ONU-SGA, 2011), corresponde a la categoría 4 (DL50 se ubica entre 300-2000 mg/Kg).

The Lippia alba (alba L.) of the Verbenaceae family, popularly known as “Cidrón” or “pennyroyal”; depending on your geographic location, soil which develops and tissue or organ, presents essential oils in its composition, which are used in natural and traditional medicine for the treatment of digestive, respiratory, skin diseases and sedative disorders through infusion or decoction of leaves, aerial parts or flowers. So it has a high therapeutic potential, being important to detect possible toxic effects post administration. Therefore, the aim of the study was to evaluate the chemical composition and oral acute toxicity of essential oil of L. alba single dose and repeat dose. the essential oil of the leaves and aerial parts extracted by steam stripping using a Clevenger trap and analyzed by gas chromatography coupled to mass detector. Toxicity was evaluated using NRMI mice of both sexes and OECD Guidelines 423 and 407. 34 constituents of the essential oil, the majority were were obtained: geranialdehyde (45.30%), neral (30.23%), beta-farnesol (7.78%), caryophyllene (2.85%), azulene (2.52%) and myrcene beta (2.38%). (OECD Guidelines 423) were evident signs and symptoms of neurological damage and moderate engine (900 mg / kg of essential oil) with 100% survival and severe (1500 mg / kg) lethal for males and with 2000 mg / kg all the animals died. It is concluded that in the essential oil predominated geranialdehyde and neral constituents; classified as “citral” chemotypes and according to the Classification of toxicity Globally Harmonized System of Classification and Labelling of Chemicals (UN GHS, 2011), corresponds to category 4 (LD50 is between 300-2000 mg / kg).

Óleo essencial das folhas do quimiotipo II de Lippia alba (Mill.) N. E. Brown: mecanismo deação antimicrobiano em staphylococcus aureus e atividade cicatrizante

A Lippia alba é uma planta medicinal muito utilizada pela população brasileira para o tratamento de diversas doenças. A composição química de seu óleo essencial apresent avariação quantitativa e qualitativa, classificando em quimiotipos. No Ceará, o quimiotipo II é reconhecido pelo óleo essencial de L. alba (OELaII) conter elevados teores de limoneno e citral. A cicatrização de feridas envolve eventos celulares e moleculares que interagem para que ocorra a reconstituição do tecido. A contaminação bacteriana, principalmente por S.aureus, é um dos principais fatores que afeta a cicatrização. O objetivo desse estudo foi investigar o mecanismo de ação do OELaII sobre S. aureus e sua atividade cicatrizante. O mecanismo de ação foi avaliado através de alterações da membrana, na morfologia bacterianae na inibição de fatores de virulência. A ação cicatrizante foi determinada in vivo em modelo experimental de feridas cutâneas através da área da ferida, da contração, da análise clínica,histopatológica e microbiológica. O mecanismo de ação do OELaII sobre S. aureus ocorreu por ação direta no envoltório celular comprovada pela alteração irreversível na permeabilidade da membrana citoplasmática, pois os resultados mostraram aumento dacaptação de cristal violeta, liberação de ácidos nucleicos e efluxo de potássio, bem comodanos na parede bacteriana visualizado por Microscopia de Força Atômica......

Lippia alba is a medicinal plant widely used by the population for the treatment of variousdiseases. The chemical composition of the essential oil presents quantitative and qualitativevariation, ranking chemotypes. In Ceará, the chemotype II is recognized by the essential oil ofL. alba (OELaII) contain high levels of limonene and citral. Wound healing involves cellularand molecular events that occur interact to tissue reconstitution. Bacterial contamination,mainly by S. aureus is one of the main factors that affect healing. The aim of this study was toinvestigate the mechanism of action of OELaII on S. aureus and its healing activity. Themechanism of action was evaluated through the membrane changes, in bacterial morphologyand inhibition of virulence factors. The healing action was determined in vivo in experimentalskin wounds through the area of the wound contraction, clinical analysis, histological andmicrobiological. The OELaII the mechanism of action on S. aureus occurred by direct actionin the cell envelope proven by irreversible changes in the permeability of the cytoplasmicmembrane, as the results showed an increase of crystal violet uptake, release of nucleic acidsand potassium efflux and damage the bacterial wall visualized by Atomic Force Microscopy...

Quimiotipos, Extracción, Composición y Aplicaciones del Aceite Esencial de Lippia alba / Quimiotipos, extração, composição e uso do óleo essencial de Lippia alba / Chemotypes, extraction, chemical composition and use of lippia alba essential oil

RESUMO Lippia alba é uma planta amplamente distribuída nas zonas tropicais, subtropicais e temperadas das Américas, África e Ásia. O óleo essencial de L. alba tem sido amplamente estudado, entretanto apresenta variações de produção. Portanto este estudo teve como objetivo realizar uma revisão dos principais quimiotipos, métodos de extração, composição e aplicação do óleo essencial de L. alba. Neste estudo são discutidos os principais quimiotipos e sua relação com fatores genéticos e características morfológicas. Também são discutidos os fatores que afetam o rendimento de produção, composição química, métodos de extração e do uso e da atividade biológica do óleo essencial de L. alba. Apesar da vasta literatura sobre os óleos essenciais de L. alba, ainda desenvolvimento de aplicações para a produção de cosméticos, fármacos e alimentos, bem como faltam definições agronomicas sobre o cultivo e melhoramento desta planta.

ABSTRACT Lippia alba is a plant widely distributed in tropical, subtropical and temperate zones of the Americas, Africa and Asia. The essential oil of L. alba has been widely studied and there are many variations in the production process. Therefore, this study is aimed at conducting a review of the main chemotypes, extraction methods, composition and application of the essential oil of L. alba. In this study, the main chemotypes and its relation to genetic and morphological characteristics are discussed. It also discusses the factors that affect the yield, chemical composition, extraction methods and the use and the biological activity of the essential oil of L. alba. Despite the vast literature on the essential oils of L. alba, there is still a lack of development in its application for the production of cosmetics, pharmaceuticals and food, as well as a lack of agronomic definitions for its cultivation and genetic improvement.

Atividade antagonista do óleo essencial de Lippia alba (Mill. ) N. E. Brown (Verbenaceae) sobre Meloidogyne incognita (Kofoid & White) Chitwood / Antagonist activity of the essential oil Lippia alba (Mill. ) N. E. Brown (Verbenaceae) on Meloidogyne incognita (Kofoid & White) Chitwood

RESUMO O controle de fitonematóides é uma tarefa difícil. A alta infestação de nematoides no solo obriga os produtores a usarem doses mais elevadas de nematicidas, ou ainda, a aumentarem a freqüência das aplicações, geralmente no solo, ocasionando maior potencial de dano ao homem e ao ambiente. Além disso, nematicidas convencionais vêm sofrendo grandes restrições de uso em muitos países. Desse modo, a busca de novas medidas de controle de fitonematóides é uma prioridade da agricultura sustentável. Desta forma, objetivou-se estudar o efeito de óleos essenciais de Lippia Alba, na mortalidade de juvenis pré-parasitas do segundo estádio de Meloidogyne incognita. O ensaio foi montado em delineamento inteiramente casualizado, com 4 repetições, em esquema fatorial 3×4 (3 quimiotipos x 4 concentrações) empregando-se três diferentes quimiotipos de L. Alba (I, II, III) nas concentrações de 0, 100, 500 e 1000 ppm, coletados em diferentes horas do dia (7, 9, 13, 16 e 19h). Os dados coletados (% de mortalidade) foram submetidos à análise de variância e os valores médios comparados por meio do teste de Tukey. Foi possível concluir que Lippia alba contém, em seu óleo essencial, compostos com efeitos significativos na mortalidade de juvenis de segundo estádio (J2) de M. incognita. Dessa forma, é importante destacar que o acentuado efeito nematicida do óleo essencial de L. alba sugere a possibilidade de seu uso no controle de M. incognita.

ABSTRACT The control of nematodes is difficult. The high infestation of nematodes in the soil requires the producers use higher doses of nematicides, increasing the frequency of applications, usually in soil, causing negative effects to humans and the environment. Furthermore, conventional nematicides have suffered great use restrictions in many countries. Thus, the search for new measures to control nematodes is a priority of sustainable agriculture. Thus, the objective of this study was available the effect of essential oils from Lippia Alba against Meloidogyne incognita. The experiment was conducted in a completely randomized design with four replications in a factorial 3x4 (3 chemotypes and 4 concentrations) using three different chemotypes of L. Alba (I, II, III) at concentrations of 0, 100, 500 and 1000 ppm, collected in different times of day (7 AM, 9 AM, 1 PM, 4 PM e 7 PM). The data collected (% mortality) were subjected to analysis of variance and mean values were compared using the Tukey test. It was able to conclude that L. alba contains in its essential oil, compounds with significant effects on mortality of M. incognita. Thus, it is important to note that the greatest nematicide effect of essential oil of L. alba suggests the possibility of its use to control M. incognita.

Essential oil of Lippia alba and its main constituent citral block the excitability of rat sciatic nerves

Lippia alba is empirically used for infusions, teas, macerates, and hydroalcoholic extracts because of its antispasmodic, analgesic, sedative, and anxiolytic effects. Citral is a mixture of trans-geranial and cis-neral and is the main constituent of L. alba essential oil and possesses analgesic, anxiolytic, anticonvulsant, and sedative effects. The present study evaluated the effects of the essential oil of L. alba (EOLa) and citral on compound action potentials (CAPs) in Wistar rat sciatic nerves. Both drugs inhibited CAP in a concentration-dependent manner. The calculated half-maximal inhibitory concentrations (IC50) of peak-to-peak amplitude were 53.2 µg/mL and 35.00 µg/mL (or 230 µM) for EOLa and citral, respectively. Peak-to-peak amplitude of the CAP was significantly reduced by 30 µg/mL EOLa and 10 µg/mL citral. EOLa and citral (at 60 and 30 µg/mL, values close to their respective IC50 for CAP blockade) significantly increased chronaxy and rheobase. The conduction velocity of the first and second CAP components was statistically reduced to ∼86% of control with 10 µg/mL EOLa and ∼90% of control with 3 µg/mL citral. This study showed that EOLa inhibited nerve excitability and this effect can be explained by the presence of citral in its composition. Both EOLa and citral showed inhibitory actions at lower concentrations compared with other essential oils and constituents with local anesthetic activity. In conclusion, these data demonstrate that EOLa and citral are promising agents in the development of new drugs with local anesthetic activity.

Estabilidade lipídica de filés de carpa húngara congelados tratados com extratos de Lippia Alba / Lipid stability of frozen common carp fillets treated with Lippia alba extract

O objetivo do presente estudo foi avaliar o efeito da aspersão de extratos de Lippia alba na estabilidade lipídica de filés de carpa húngara armazenados a -18±2°C. Filés não tratados (controle sem aspersão) ou aspergidos (1mL 10g-1 de filé) com água destilada (controle água destilada) ou com extratos de L. alba (0,10g mL-1) hidrometanólico ou aquoso foram analisados durante o armazenamento nos dias zero, 90 e 180. Independente do tempo de congelamento, o extrato hidrometanólico reduziu os valores de dienos conjugados (DC) dos filés em relação aos demais tratamentos, além de reduzir os valores de ácidos graxos livres aos 90 dias (P<0,05). O extrato aquoso resultou em maior teor de peróxidos após 180 dias de congelamento comparado aos demais tratamentos (P<0,05). Os extratos hidrometanólico e aquoso reduziram os valores de substâncias reativas ao ácido tiobarbitúrico (TBARS) após 180 dias, comparados aos filés tratados com água destilada e sem aspersão (P<0,05). Ambos os extratos de L. alba retardaram a oxidação lipídica, sendo que o extrato aquoso retardou a degradação de produtos primários da oxidação lipídica (peróxidos) em produtos secundários (TBARS), enquanto o extrato hidrometanólico parece ser mais eficiente, pois inibiu de forma similar tanto a formação de DC e peróxidos, quanto a sua degradação em produtos secundários (TBARS).

The effect of sprinkling with Lippia alba extracts was evaluated on the lipid stability of common carp fillets stored at -18±2°C. Fillets that received no treatment (no spray control) or that were sprayed (1mL 10g-1 fillet) with distilled water (water control) or with hydro-methanolic or aqueous extract of L. alba (0.10g mL-1) were evaluated immediately (time zero) and after 90 and 180 days. Regardless of the storage time, the hydro-methanolic extract reduced the conjugated dienes (CD) values of fillets compared to the other treatments, and reduced the free fatty acid levels at 90 days (P<0.05). The aqueous extract caused higher peroxide value after 180 days of frozen storage compared to the other treatments (P<0.05). The hydro-methanolic and aqueous extracts reduced thiobarbituric acid reactive substances (TBARS) values after 180 days compared to the non-treated fillets or to the water-sprayed fillets (P<0.05). Both extracts have delayed lipid oxidation. While the aqueous L. alba extract delayed the degradation of primary oxidation products (peroxides) into secondary products (TBARS), the hydro-methanolic extract was more efficient as it inhibited both the CD and peroxide formation and its degradation into secondary products (TBARS).

Bioconcentraciones foliares de elementos minerales en Lippia alba (salvia morada) / Foliar bioconcentrations of mineral elements in Lippia alba (purple salvia)

Introducción: la salvia morada es un subarbusto aromático, que crece de modo espontáneo en América Central y del Sur. Frecuentemente es cultivada en jardines como ornamental, por su intenso aroma y sus propiedades medicinales y culinarias. No se encontraron estudios sobre parámetros nutricionales en esta especie. Objetivos: determinar las bioconcentraciones foliares de elementos minerales en Lippia alba (Mill.) N. E. Br. ex Britton & P. Wilson que crece naturalmente en el bioambiente del norte de la provincia de Corrientes. Métodos: se evaluaron muestras foliares de plantas de Lippia alba de más de 2 años de implantación. Las variables analizadas fueron las concentraciones foliares de nitrógeno, fósforo, potasio, azufre, cinc, cobre, hierro y manganeso. La determinación de nitrógeno se realizó mediante el método de Kjeldhal; el fósforo y azufre por espectrometría de absorción molecular, y el resto de los elementos por espectrometría de absorción atómica. Para el análisis de los resultados se aplicaron herramientas de estadística descriptiva, mediante el programa Infostat. Resultados: las bioconcentraciones foliares de esta especie variaron entre 2,39 y 2,33 % de N; 0,2-0,4 % de P; 1,0-3,0 % de K; 0,12-0,49 % de S; 150-300 ppm de Fe, 150-300 ppm de Mn, hasta 50 ppm de Zn; y hasta 70 ppm de Cu. Las concentraciones foliares medias de nitrógeno, manganeso y cobre resultaron mayores en los meses de primavera y verano decreciendo en los meses de otoño e invierno. Las concentraciones foliares medias de fósforo y potasio se presentaron constantes durante los 3 años. El azufre, el hierro y el cinc, se acumularon en las hojas, especialmente en los meses de invierno. Conclusiones: se observaron variaciones estacionales de las concentraciones foliares medias estudiadas.

Introduction: purple salvia is an aromatic shrub that grows wild in Central and South America. It is often found in gardens for ornamental purposes, due to its strong fragrance and medicinal and culinary properties. No studies were found about nutritional parameters for this species. Objectives: determine the foliar bioconcentrations of mineral elements in Lippia alba (Mill.) N. E. Br. ex Britton & P. Wilson, naturally growing in the bio-environment of the north of Corrientes province. Methods: an evaluation was conducted of foliar samples of Lippia alba of more than 2 years after planting. The variables analyzed were foliar concentrations of nitrogen, phosphorus, potassium, sulfur, zinc, copper, iron and manganese. Nitrogen was determined by the Kjeldahl method, phosphorus and sulfur by molecular absorption spectrometry, and the rest of the elements by atomic absorption spectrometry. Descriptive statistics tools were used for the analysis of results, which was based on the InfoStat program. Results: foliar bioconcentrations ranged between 2.39 and 2.33 % N, 0.2-0.4 % P, 1.0-3.0 % K, 0.12-0.49 % S, 150-300 ppm Fe, 150-300 ppm Mn, up to 50 ppm Zn, and up to 70 ppm Cu. Mean foliar concentrations of nitrogen, manganese and copper were higher in spring and summer months, and lower in autumn and winter months. Mean foliar concentrations of phosphorus and potassium remained stable throughout the 3 years. Concentration of sulfur, iron and zinc in leaves was higher in winter months. Conclusions: seasonal variations were found in the mean foliar concentrations studied.

Lippia alba morphotypes cidreira and melissa exhibit significant differences in leaf characteristics and essential oil profile

Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson, Verbenaceae, is widely used in traditional Brazilian medicine for the treatment of abdominal distress. The species exhibits considerable chemical and morphological diversity, and various chemotypes have been characterized. A comparative study of L. alba, has been carried out of the morphoanatomical characteristics of the leaves and the profiles of the essential oils of the morphotypes cidreira and melissa grown in the Medicinal Plant Garden of the Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil. The mean plant height of cidreira was 1.80 m and the stems and branches were fairly erect, while melissa plants were smaller (1.60 m) and presented prostrate stems and branches. Although the leaf of the morphotypes look were similar, the mean values of length, width and area of the leaves of cidreira (respectively, 7.42 cm, 3.32 cm and 17.31 cm²) differed significantly from those of melissa (4.68 cm, 2.35 cm and 7.32 cm2). The morphotypes presented amphistomatic leaves with uniseriate epidermis on both surfaces. The mesophyll was dorsiventral, but in cidreira the palisade parenchyma was biseriate while in melissa it was uniseriate. Simple tector and capitate glandular trichomes were present on the adaxial and abaxial surfaces of the leaf blades of both morphotypes. Six distinct types of glandular trichomes could be distinguished: types I and II were present in both morphotypes, while type III was detected only in cidreira, and types IV to VI were present only in melissa. The two morphotypes also differed with respect to the composition of the essential oil, cidreira produced oil composed mainly of citral, while the oil from melissa was rich in citral, limonene and carvone.

Influência do horário de colheita sobre o rendimento e composição do óleo essencial de erva-cidreira brasileira [Lippia alba (Mill. ) N. E. Br. ] / Influence of harvest time on the yield and composition of essential oil from the Brazilian "erva-cidreira" [Lippia alba (Mill. ) N. E. Br. ]

Este trabalho teve como objetivo verificar se o horário de colheita da erva-cidreira brasileira [Lippia alba (Mill.) N. E. Br.], fenotipo carvona-limoneno, tem influência sobre a produção de massa foliar, rendimento e composição do óleo essencial. Foram avaliados cinco horários de colheita quando a cultura estava com 145 dias desde o transplante: 8:00, 10:00, 12:00, 14:00 e 16:00 h com cinco repetições, distribuídos em blocos casualizados. O experimento foi conduzido na Fazenda Experimental Lageado da FCA-UNESP/Botucatu no Setor de Horticultura do Departamento de Produção Vegetal. A colheita foi realizada a 15 cm da superfície do solo e o óleo essencial obtido através de hidrodestilação, em aparelho tipo Clevenger. Levou-se em consideração os fatores agronômicos e o rendimento obtido por hidrodestilação. O óleo essencial foi analisado em cromatógrafo gasoso acoplado a espectrômetro de massas (CG/EM). Os resultados foram submetidos à análise de variância (Teste F) e as médias comparadas pelo teste de Tukey. Não houve diferença estatística para produção de massa foliar, teor de óleo essencial e produtividade de óleo essencial em massa fresca e seca. No entanto, entre os compostos majoritários do óleo essencial das folhas, carvona e limoneno, a melhor produtividade de carvona foi obtida às 10:00 h, em matéria fresca (2,050 L ha-1) e em matéria seca (2,068 L ha-1), e para o limoneno às 16:00 h, em matéria fresca (1,068 L ha-1) e em matéria seca (1,060 L ha-1).

This study aimed to verify whether the harvest time of the "Brazilian erva-cidreira" [Lippia alba (Mill.) N. E. Br.], limonene-carvone chemotype, influences leaf mass production and essential oil yield and composition. Five different harvest times were evaluated at 145 days after transplanting: 8:00, 10:00, 12:00, 14:00 and 16:00, with five replicates, distributed in randomized blocks. The experiment was carried out at Lageado Experimental Farm of the School of Agronomical Sciences FCA-UNESP/Botucatu, Horticulture Sector, Department of Plant Production. Harvest was conducted from 15 cm above the soil surface and essential oil was obtained through steam distillation in a Clevenger-type apparatus. Both the agronomical factors and the yield obtained through steam distillation were considered. The essential oil was analyzed in a gas chromatographer coupled to a mass spectrometer (GC/MS). Results were subjected to analysis of variance (F Test) and means were compared according to Tukey's test. There was no statistical difference for leaf mass production, essential oil content and essential oil yield in fresh and dry matter. However, considering the major essential oil compounds, carvone and limonene, the best yield was obtained at 10:00 for carvone in fresh (2050 L ha-1) and dry matter (2068 L ha-1), and at 16:00 for limonene in fresh (1068 L ha-1) and dry matter (1060 L ha-1).

Oil production at different stages of leaf development in Lippia alba

The aim of this work was to analyze terpene oil production and terpene synthases (TPS) gene expression from leaves at different developmental stages of different chemotypes of Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson, Verbenaceae. Hydro-distilled essential oil were used for chemical analysis and gene expression of three monoterpene synthase genes called LaTPS12, LaTPS23 and LaTPS25 were used for analyses of gene expression associated to oil production. The putative genes were associated to TPS-b gene class. Semi-quantitative PCR and quantitative PCR (qPCR) analysis were used to investigate the expression profile of those three putative genes in different leaf stages and different chemotypes. Additionally, total oil production and gene expression of putative TPS genes cloned from L. alba chemotype linalool were evaluated at different stages of leaf development. The expression level of those three genes was higher when the highest oil production was observed, mainly in young leaves at the fourth nodal segment for all evaluated chemotypes. Total oil production was higher at leaves that had unopened trichomes. We also observed that the 1mM of MeJA treatment increased the gene expression in all chemotypes after 24 h elicitation.

Anxiolytic effects of repeated treatment with an essential oil from Lippia alba and (R)-(-)-carvone in the elevated T-maze

Lippia alba (Mill.) N.E. Brown (Verbenaceae) is widely used in different regions of Central and South America as a tranquilizer. The plant’s anxiolytic properties, however, merit investigation. The present study evaluated the effects of repeated daily (14 days) intraperitoneal (ip) treatment with an essential oil (EO) from a chemotype of L. alba (LA, chemotype II, 12.5 and 25 mg/kg; N = 6-8) and (R)-(-)-carvone (25 mg/kg; N = 8-12), the main constituent of this chemotype, on male Wistar rats (weighing 250 g at the beginning of the experiments) submitted to the elevated T-maze (ETM). The ETM allows the measurement of two defensive responses: inhibitory avoidance and one-way escape. In terms of psychopathology, these responses have been related to generalized anxiety and panic disorder, respectively. Treatment with the EO impaired ETM avoidance latencies, without altering escape, in a way similar to the reference drug diazepam (P < 0.05) (avoidance 2: control = 84.6 ± 35.2; EO 12.5 mg/kg = 11.8 ± 3.8; EO 25 mg/kg = 14.6 ± 2.7; diazepam = 7 ± 2.1). (R)-(-)-carvone also significantly altered this same response (P < 0.05; avoidance 1: control = 91.9 ± 31.5; carvone = 11.6 ± 1.8; diazepam = 8.1 ± 3.3). These results were not due to motor changes since no significant effects were detected in an open field. These observations suggest that LA exerts anxiolytic-like effects on a specific subset of defensive behaviors that have been implicated in generalized anxiety disorder, and suggest that carvone is one of the constituents of LA responsible for its action as a tranquilizer.

Tipos de estacas e diferentes substratos na propagação vegetativa da erva cidreira (quimiotipos I, II e III) / Types of cuttings and different substrates in the vegetative propagation of the Lippia alba (chemotypes I, II and III)

A erva-cidreira [Lippia alba (Mill.) N. E. Brown] é comumente utilizada no Brasil como planta medicinal por suas propriedades analgésica, antiespasmódica, calmante e sedativa. O objetivo do trabalho foi avaliar o efeito do tipo de estaca e de diferentes substratos sobre a propagação vegetativa da erva-cidreira quimiotipos I, II e III. O material botânico utilizado foi estacas de erva-cidreira quimiotipos I, II, e III, com aproximadamente 12 cm de comprimento. No primeiro experimento, foram utilizadas estacas (apicais e basais) dos três quimiotipos, em um esquema fatorial 2 x 3. No segundo experimento, as estacas dos três quimiotipos foram propagadas em três substratos: substrato comercial + esterco bovino curtido (na proporção de 1:1); substrato comercial + palha de arroz carbonizada (na proporção de 1:1); e substrato comercial, em um esquema fatorial de 3 x 3 com 4 repetições, onde se determinou a massa fresca e seca da raiz, caule e folhas. Estacas apicais e basais podem ser utilizadas na propagação vegetativa da erva-cidreira quimiotipos I, II e III. As misturas substrato comercial + palha de arroz carbonizada e substrato comercial + esterco bovino curtido são recomendadas para a propagação vegetativa da erva-cidreira quimiotipos I, II e III.

The Lippia alba is commonly used in Brazil as a medicinal plant for its analgesic properties, antispasmodic, calming and sedative. The objective of this study was to evaluate the effect of cutting type and different substrates on the propagation of the Lippia alba chemotypes I, II and III. The botanical material used was stakes Lippia alba chemotypes I, II and III, with approximately 12 cm in length. In the first experiment, we used cuttings (apical and basal) of the three chemotypes in a 2 x 3 factorial arrangement. In the second experiment, cuttings of three chemotypes were propagated in three substrates: commercial substrate + cattle manure (1:1), commercial substrate + carbonized rice hulls (1:1) and commercial substrate, in a factorial 3 x 3 with 4 replicates, was determined fresh and dry weight of root, stem and leaves. Apical and basal cuttings can be used in the propagation of the Lippia alba chemotypes I, II and III. The commercial substrate mixtures + carbonized rice hulls and commercial substrate + cattle manure are recommended for the propagation of lemongrass chemotypes I, II and III.