Screening of essential oils against oxacillin - resistant Staphylococcus aureus strains isolated from bovine mastitis / Screening de aceites esenciales frente a Staphylococcus aureus oxacilino - resistentes extraidos de aislados de mastitis bovina

Bovine mastitis is a disease wi th far - reaching consequences for the dairy industry. Staphylococcus aureus is a pathogen that is especially resistant to antibiotics. The objective of this study was to evaluate the antimicrobial activity of the essential oils Lippia citriodora (Lam.), Thy mus vulgaris (L), and a mixture of the essential oils Lippia citriodora and Thymus vulgaris (50/50 v/v), against isolates of oxacillin - resistant Staphylococcus aureus (n=15) of positive cases of bovine mastitis. For the statistical analysis, the IBM SPSS s tatistical package was used. The mixture of essential oils ( Lippia citriodora and Thymus vulgaris (50/50 v/v)) obtained the most significant antimicrobial activity in relation to pure essential oils. It is therefore concluded that the mixture of these oils boosts their antimicrobial activity ( p <0.05). The minimum inhibitory and bactericidal concentration of this mixture for the total isolations was 12 µL/L and 25 µL/mL, respectively.

La mastitis bovina es una enfermedad de gran impacto para la industria lechera. El Staphylococcus aureus es uno de los principales patógenos, especialmente aquellos resistentes a los antibióticos. El objetivo de este estudio fue evaluar la actividad antimicrobiana de los aceites esenciales de Lippia citriodora (Lam.), Thymus vulgaris (L), y una mezcla de aceites esenciales de Lippia citriodora y Thymus vulgaris (50/50 v/v), frente a aislamientos clínicos de Staph ylococcus aureus oxacilino - resistentes (n=15) de mastitis bovina. Se utilizó p rograma estadístico IBM SPSS y se concluyó la diferencia significativa a un p <0.05. La mezcla de aceites esenciales ( Lippia citriodora y Thymus vulgaris (50/50 v/v)), obtuvo la m ayor actividad antimicrobiana en relación a los aceites esenciales puros, se concluye que la mezcla de estos aceites potencia su actividad antimicrobiana ( p <0.019). La concentración mínima inhibitoria y bactericida de esta mezcla fue del 12 µL/mL y 25 µL/m L, respectivamente, y puede ser una alternativa terapéutica.

Efecto ecotóxico y preventivo del uso de aceites esenciales en la mastitis bovina / Ecotoxic and preventive effect of the use of essential oils in bovine mastitis

The essential oils of Lippia citriodora (Ort.) and Lippia origanoides (Kunth) have shown antimicrobial activity associated with mastitis. The objective of this study was to evaluate its ecotoxic effect with the Artemia salina bioassay and the prevention of mastitis through an in vivo test in cattle (n=20) with a product based on these oils using a conventional product as a control. Contact hypersensitivity, the effect on somatic cells, and residuality in mil k samples were evaluated. The results of the Artemia salina bioassay were 10.05 and 19.36 (µg/mL) respectively. No negative effects or contact hypersensitivity were observed, and no residual metabolites were found in post - test milk. The somatic cell count showed 75% effectiveness in the prevention of mastitis with essential oils compared to 62.5% with the conventional product. The evaluated formulation could be used in the prevention of bovine mastitis safely, further investigation is required.

Los aceites esenciales de Lippia citriodora (Ort.) y Lippia origanoides (Kunth), han mostrado acti vidad antimicrobiana asociada a la mastitis. El objetivo de este estudio fue evaluar su efecto ecotóxico con el bioensayo Artemia salina y la prevención de mastitis mediante un ensayo in vivo en bovinos (n=20) con un producto a base de estos aceites utiliz ando como control un producto convencional. Se evaluó la hipersensibilidad de contacto, efecto en células somáticas y residualidad en muestra de leche. Los resultados del bioensayo de Artemia salina fueron 10,05 y 19,36 (µg/mL) respectivamente. No se obser varon efectos negativos, ni hipersensibilidad de contacto, y no se encontraron metabolitos residuales en leche posterior al ensayo. El conteo de células somáticas mostró efectividad en la prevención de mastitis del 75% con aceites esenciales frente al 62.5 % del producto convencional. La formulación evaluada podría ser utilizada en la prevención de la mastitis bovina de forma segura, se requiere profundizar en la investigación.

Biotechnological potential of essential oils from different chemotypes of Lippia alba(Mill.) N.E.Br. ex Britton & P. Wilson / Potencial biotecnológico de los aceites esenciales de diferentes quimiotipos de Lippia alba(Mill.) N.E.Br. ex Britton y P. Wilson

This study reports the biotechnological importance of seven Lippia albaspecimens collected in different places in Brazil, and evaluation of some activities as larvicidal against Aedes spp., antifungal against dermatophytes; cytotoxicity against SNB-19 (astrocytoma), HCT-116 (human colon) and PC-3 (human prostate) cancer cell lines, and inhibition of the enzyme acetylcholinesterase (AChE). The essential oils, whose extraction yield was about 1.24 w/w ± 0.9%, showed differences in their chemical composition and considered as chemotypes. The essential oils containing neral and geranial as main constituents showed better action against HCT-116 cell lines (IC50 value was 9.22 µg/mL), larvicidal activity against arbovirus vectors (LC50 value against A. aegypti was 1.59 µg/mL) and inhibition of AChE (halo inhibition zone was 1 cm). The essential oils containing mainly monoterpenoids showed better antifungal action with MIC values range from 0.15 to 1.25 mg/mL. This chemical and biological characterization may be useful for biotechnological applications.

Este estudio reporta la importancia biotecnológica de siete especímenes de Lippia alba recolectados en diferentes lugares de Brasil, y evaluación de algunas actividades como larvicida contra Aedes spp., Antifúngico contra dermatofitos; citotoxicidad contra líneas celulares de cáncer SNB-19 (astrocitoma), HCT-116 (colon humano) y PC-3 (próstata humana), e inhibición de la enzima acetilcolinesterasa (AChE). Los aceites esenciales, cuyo rendimiento de extracción fue de aproximadamente 1,24 p/p ± 0,9%, mostraron diferencias en su composición química y se consideraron quimiotipos. Los aceites esenciales que contienen neral y geranial como constituyentes principales mostraron una mejor acción contra las líneas celulares HCT-116 (el valor de CI50 fue de 9.22 µg/mL), actividad larvicida contra los vectores de arbovirus (el valor de LC50 contra A. aegypti fue de 1.59 µg/mL) e inhibición de AChE (la zona de inhibición del halo era de 1 cm). Los aceites esenciales que contienen principalmente monoterpenoides mostraron una mejor acción antifúngica con valores de CMI en el rango de 0.15 a 1.25 mg/mL. Esta caracterización química y biológica puede ser útil para aplicaciones biotecnológicas.

Seasonal variation of essential oil of germplasm of Lippia origanoidesKunth. (Verbenaceae) / Variación estacional de aceite esencial de germoplasma de Lippia origanoidesKunth. (Verbenaceae)

The biological activities attributed to Lippia origanoides Kunth. vary according to the chemical composition of its essential oil, which can be related to weather factors. The aim of this research was the seasonal study of the essential oil chemical composition taken from L. origanoides accessions maintained in germplasm bank in vivo. The essential oil was extracted by hydrodistillation in Clevenger apparatus and identification of the components was done by GC-MS. We have identified 15 compounds of which carvacrol, p-cymene, thymol and methyl-ether-thymol were majority throughout the year for most accessions. In three accessions showed fluctuations in the production of the major compound during the year like the α-pinene, limonene and borneol. The essential oil showed variation in chemical composition throughout the year, the major compound in most of the accessions during the year is carvacrol. In some accessions, there are alterations of the majority component.

Las actividades biológicas atribuidas a Lippia origanoides Kunth. varían según la composición química de su aceite esencial, lo que puede estar relacionado con factores climáticos. El objetivo de esta investigación fue el estudio estacional de la composición química del aceite esencial extraído de accesiones de L. origanoides mantenidas en banco de germoplasma in vivo. El aceite esencial se extrajo mediante hidrodestilación en un aparato Clevenger y la identificación de los componentes se realizó mediante GC-MS. Hemos identificado 15 compuestos de los cuales carvacrol, p-cimeno, timol y metil-éter-timol fueron mayoritarios durante todo el año para la mayoría de las accesiones. En tres accesiones se observaron fluctuaciones en la producción del compuesto principal durante el año como el α-pineno, limoneno y borneol. El aceite esencial mostró variación en la composición química a lo largo del año, el compuesto principal en la mayoría de las accesiones durante el año es el carvacrol. En algunas accesiones, hay alteraciones del componente mayoritario.

Plantas com ação no sistema nervoso central que constam na relação nacional de plantas medicinais de interesse ao sus (RENISUS) / Plants acting on the central nervous system: review plantas com ação no sistema nervoso central que constam na relação nacional de plantas medicinais de interesse ao sus (RENISUS) / Plantas con acción sobre el sistema nervioso central incluidas en la lista nacional de plantas medicinales de interés para sus (RENISUS)

A depressão é uma doença grave que atinge a população em geral, estudos epidemiológicos estimam que a prevalência da depressão ao longo da vida no Brasil está em torno de 15,5%. Os fatores que desencadeiam o aparecimento da depressão incluem fatores sociais, psicológicos, biológicos e também fatores externos específicos como eventos estressantes, solidão, consumo de álcool e drogas, doenças crônicas e dar á luz (depressão pós-parto). O objetivo da presente pesquisa consistiu em realizar uma revisão bibliográfica sobre as principais plantas medicinais com ação antidepressiva. A ansiedade vem se tornando um dos principais problemas da atualidade, sendo intensificada pela pandemia causada pelo coronavírus, onde constatou-se que durante o pico da pandemia onde os casos confirmados de COVID-19 no Brasil ascenderam de 45.757 para 330.890, e as mortes, de 2.906 para 21.048, o sentimento de tristeza/depressão atingiu 40% dos adultos brasileiros. Os sintomas de depressão podem ser amenizados quando a disponibilidade sináptica de monoaminas são aumentadas, e esse aumento pode ocorrer através da diminuição da metabolização desses neurotransmissores. Neste sentido, busca-se através da farmacoterapia a utilização de antidepressivos que disponibilizem as monoaminas na fenda sináptica. A escolha do fármaco é feita com base nos sintomas da depressão e na boa resposta a uma determinada classe de antidepressivos. Em fevereiro de 2009 o Ministério da saúde lançou a Relação Nacional de Plantas Medicinais de Interesse ao SUS (RENISUS), contendo 71 espécies vegetais que são distribuídas de forma in natura nas unidades básicas de saúde (UBS). Destas, somente três espécies apresentam efeito antidepressivo e ansiolítico comprovados na literatura sendo Matricharia chamomilla, Erytrinum mulungu e a Passiflora incarnata que também fazem parte da RENISUS. Além destas, outras espécies como a Melissa officinalis, Lippia alba, Valeriana officinalis e Piper methysticum são utilizadas pela população para tratar ansiedade, insônia e depressão, sugerindo desta forma que estas espécies sejam incluídas na RENISUS.

Depression is a serious disease that affects the general population, epidemiological studies estimate that the prevalence of depression throughout life in Brazil is around 15.5%. The factors that trigger the onset of depression include social, psychological, biological and also specific external factors such as stressful events, loneliness, alcohol and drug consumption, chronic diseases and giving birth (postpartum depression). The objective of the present research was to carry out a literature review on the main medicinal plants with antidepressant action. Anxiety has become one of the main problems of today, being intensified by the pandemic caused by the coronavirus, where it was found that during the peak of the pandemic where confirmed cases of COVID-19 in Brazil rose from 45,757 to 330,890, and deaths, from 2,906 to 21,048, the feeling of sadness/depression reached 40% of Brazilian adults. Symptoms of depression can be alleviated when synaptic availability of monoamines is increased, and this increase can occur through decreased metabolization of these neurotransmitters. In this sense, the use of antidepressants that make monoamines available in the synaptic cleft is sought through pharmacotherapy. The choice of drug is based on symptoms of depression and good response to a particular class of antidepressants. In February 2009, the Ministry of Health launched the National List of Medicinal Plants of Interest to the SUS (RENISUS), containing 71 plant species that are distributed in natura form in basic health units (UBS). Of these, only three species have antidepressant and anxiolytic effects proven in the literature, being Matricharia chamomilla, Erytrinum mulungu and Passiflora incarnata, which are also part of RENISUS. In addition to these, other species such as Melissa officinalis, Lippia alba, Valeriana officinalis and Piper methysticum are used by the population to treat anxiety, insomnia and depression, thus suggesting that these species are included in RENISUS.

Los estudios epidemiológicos estiman que la prevalencia de la depresión a lo largo de la vida en Brasil es de alrededor del 15,5%. Los factores que desencadenan la aparición de la depresión son sociales, psicológicos, biológicos y también factores externos específicos, como los acontecimientos estresantes, la soledad, el consumo de alcohol y drogas, las enfermedades crónicas y el parto (depresión posparto). El objetivo de esta investigación fue realizar una revisión bibliográfica sobre las principales plantas medicinales con acción antidepresiva. La ansiedad se ha convertido en uno de los principales problemas de la actualidad, intensificándose por la pandemia causada por el coronavirus, donde se encontró que durante el pico de la pandemia donde los casos confirmados de COVID-19 en Brasil aumentaron de 45.757 a 330.890, y las muertes, de 2.906 a 21.048, el sentimiento de tristeza/depresión alcanzó el 40% de los adultos brasileños. Los síntomas de la depresión pueden aliviarse cuando se aumenta la disponibilidad sináptica de las monoaminas, y este aumento puede producirse mediante una disminución de la metabolización de estos neurotransmisores. En este sentido, se busca a través de la farmacoterapia el uso de antidepresivos que hagan disponibles las monoaminas en la hendidura sináptica. La elección del fármaco se hace en función de los síntomas de la depresión y de la buena respuesta a una clase concreta de antidepresivos. En febrero de 2009, el Ministerio de Salud lanzó la Lista Nacional de Plantas Medicinales de Interés para el SUS (RENISUS), que contiene 71 especies de plantas que se distribuyen in natura en unidades básicas de salud (UBS). De ellas, sólo tres especies tienen efectos antidepresivos y ansiolíticos probados en la literatura: Matricharia chamomilla, Erytrinum mulungu y Passiflora incarnata, que también forman parte del RENISUS. Además de éstas, otras especies como Melissa officinalis, Lippia alba, Valeriana officinalis y Piper methysticum son utilizadas por la población para tratar la ansiedad, el insomnio y la depresión, lo que sugiere que estas especies se incluyan en el RENISUS.

Cytotoxic activity of ethanolic extracts of Lippia graveolens HBK leaves and stem against lung cancer cell line SK-LU-1 / Actividad citotóxica de extractos etanólicos de hoja y tallos de Lippia graveolens HBK contra la línea celular de cáncer de pulmón SK-LU-1

Lippia graveolens HBK (Verbenaceae) is an aromatic herb of economic importance in Mexico, known as oregano. The leaves are used as condiments, and people use this species for respiratory and digestive disorders. The aim of this work was to evaluate the cytotoxic effect of ethanolic extracts obtained from free-oil leaves (L9) and stem (S15), against lung cancer cell line SK-LU-1, through tetrazolium salt (MTT) assay. Extracts concentrations of 0.3 to 300 µg/mL were used and HFF-1 as normal control cells. Both L9 and S15 extracts, showed cytotoxic effect, although stem was stronger than leaves and without damage to normal cell control. The phenolic compounds caffeic acid and acacetin werein higher concentration in L9, whereas naringenin, taxifolin, eriodictyol, luteolin, and apigenin had higher concentrations in S15. The ethanolic extracts of L. graveolens have excellent cytotoxic activity, and have a wide possibility of use in lung cancer treatment.

Lippia graveolens HBK. (Verbenaceae) es una hierba aromática de importancia económica en México, conocida como orégano. Las hojas se usan como condimento y en medicina tradicional se utiliza para aliviar malestares respiratorios y digestivos. El objetivo de este trabajo fue evaluar el efecto citotóxico de extractos etanólicos obtenidos de hojas sin aceite (L9) y tallo (S15), sobre la línea celular de cáncer de pulmón SK-LU-1, mediante el ensayo de la sal de tetrazolium (MTT). Los extractos se aplicaron a concentraciones de 0,3 a 300 µg/mL y se utilizaron células HFF-1 como control normal. Tanto los extractos L9 como S15 mostraron efecto citotóxico, aunque el efecto del tallo fue mayor al de las hojas y sin daño al control celular normal. Los compuestos fenólicos ácido cafeico y acacetina se encontraron en mayor concentración en L9, mientras que naringenina, taxifolina, eriodictyol, luteolina y apigenina tuvieron mayor concentración en S15. Los extractos etanólicos de L. graveolens tienen una excelente actividad citotóxica, con amplia posibilidad de utilizar en el tratamiento de cáncer de pulmón.

Comportamiento fenológico ex situ de Lippia integrifolia "incayuyo" (Verbenaceae) de una población de base genética amplia / Ex situ phenological behavior of Lippia integrifolia "incayuyo" (Verbenaceae) from a population with a broad genetic base

Abstract: Lippia integrifolia "incayuyo" is an aromatic, sub - woody shrub used in popular medicine, aperitit drinks and compound herb s. Its choleretic, antispasmodic, biocidal, antibacterial and larvicidal activity has been proven. The objective of the work was to register the phenology of a sample of 70 genotypes from a population with a broad genetic base. The phenophases studied were : vegetative growth, flower bud, flowering and fruiting fortnightly for two years. The initiation, intensity and prolongation of the phenophases were evaluated. The moment of full bloom occurs during the second half of December. Taking this date as a refer ence, a differentiated beginning of flowering was evidenced. The results of two campaigns were compared, observing that 70% of the specimens had a similar behavor, standing out some genotypes for presenting an early flowering and longer duration. The recor ded variability suggests that much of it could be due to intrinsic factors of the plant, and therefore, feasible to be selected .

Resumen: Lippia integrifolia "incayuyo" es un arbusto aromático, subleñoso empleado en la medicina popular, bebidas aperitivas y yerbas compuestas. Se ha comprobado su actividad colerética, antiespasmódica, biocida, antibacteriana y larvicida. El objetivo del trabajo fue registrar la fenología d e una muestra de 70 genotipos de una población de base genética amplia. Las fenofases estudiadas fueron: crecimiento vegetativo, botón floral, floración y fructificación quincenalmente durante dos años. Se evaluó inicio, intensidad y prolongación de las fe nofases. El momento de plena floración ocurre durante la segunda quincena de diciembre. Tomando esta fecha como referencia, se evidenció un inicio de floración diferenciada. Se compararon los resultados de dos campañas, observando que el 70% de los ejempla res tuvo un compartimiento semejante, destacándose algunos genotipos por presentar una floración temprana y de prolongación superior. La variabilidad registrada sugiere que gran parte de ésta podría deberse a factores intrínsecos de la planta, y por ello, factible de ser seleccionados.

Inter and intra population phenotypic variability of Lippia integrifolia (Verbenaceae) and its natural situation in the west-center of Argentina / Variabilidad fenotípica inter e intra poblacional de Lippia integrifolia (Verbenaceae) y su situación natural en el centro-oeste de Argentina

The species Lippia integrifolia is an aromatic, sub-woody shrub, distributed from Northwest and Central Argentina to Bolivia. It is among the most important native aromatic species. It presents medicinal properties, some of which have been scientifically proven. As an alternative to harvesting, the process of domestication of the species was initiated. The objective of this work was to assess the phenotypic variability based on botanical-taxonomic and morpho-agronomic descriptors and the use of indicators to evaluate its natural situation in the west-center of Argentina. We worked in five wild populations, registering morphological variables, phenology, natural regeneration, herbivory and presence of diseases. L. integrifoliapresents a wide morphological variability, which can be explained with three morphological descriptors. Natural regeneration ́s rate is very low to zero, in four of the sites. The results obtained are of importance to make decisions related to the sustainable use in situ, and to begin a program of domestication of the species.

La especie Lippia integrifolia es un arbusto aromático, sub-leñoso, distribuido desde el noroeste y centro de Argentina hasta Bolivia. Es una de las especies aromáticas nativas más importantes. Presenta propiedades medicinales, algunas de las cuales han sido científicamente probadas. Como alternativa a la cosecha, se inició el proceso de domesticación de la especie. El objetivo de este trabajo fue evaluar la variabilidad fenotípica basada en descriptores botánico-taxonómicos y morfo-agronómicos y el uso de indicadores para evaluar su situación natural en el centro-oeste de Argentina. Trabajamos en cinco poblaciones silvestres, registrando variables morfológicas, fenología, regeneración natural, herviboría y presencia de enfermedades. L. integrifolia presenta una amplia variabilidad morfológica, que puede explicarse con tres descriptores morfológicos. La tasa de regeneración natural es muy baja a cero, en cuatro de los sitios. Los resultados obtenidos son importantes para tomar decisiones relacionadas con el uso sostenible in situ y para comenzar un programa de domesticación de la especie.

Effects of the healing activity of rosemary-of-Chapada (Lippia gracilis Schauer) on cutaneous lesions in rats

Purpose: To evaluate the effects of rosemary leaf essential oil-based ointments on the healing of rat skin lesions. Methods: Sixty adult male rats, with dorsal excisional skin wounds made surgically under anesthesia, were divided into three groups (n = 20): Sham group (untreated wounds); control group (CG, wounds treated with vehicle); and essential oil (EO) treated group (wounds treated with essential oil-based ointments), administered topically once daily. Skin wounds were evaluated at 4, 7, 14, and 21 days after EO or vehicle treatments. Lesions were analyzed macroscopically for the contraction degree. Formalin-fixed paraffin-embedded sections of skin wounds were used for histopathological evaluation. Results: Macroscopic evaluation showed wounds edges with thin crust without firmness and yellowish color, along with an improvement in wound contraction in EO group when compared to the other groups. A reduced inflammatory reaction, along with newly formed small diameter capillaries and more organized and elongated collagen fibers, were more frequently observed in EO group than in the other groups. Moreover, blood vessel number and collagen fibers density were significantly higher in EO group. Conclusions: Skin lesion treatment with rosemary leaf essential oil-based ointments accelerates the initial stages of healing, reduces inflammation, and increases angiogenesis, collagen fibers density, and wound contraction in rats.

Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract The composition and pharmacological properties of Lippia alba (Mill.) (L. alba) (Verbenaceae) flower and leaf essential oils (EO) were determined in this study. The major constituents in the flower EO were geranial (49.83%) and neral (32.75%), and in the leaf EO were geranial (38.06%), neral (31.02%), and limonene (18.03%). Flower EO inhibited thrombolysis induced by Bothrops moojeni (B. moojeni) and Lachesis muta muta (L. muta muta) venoms (0.05-1.2 µL mL-1). When tested against L. muta muta venom, the protective effect was smaller in both EO. The EOs prolonged the clotting time induced by L. muta muta venom and a procoagulant effect was observed on B. moojeni. In the comet assay, the flower EO presented anti-genotoxic action (damage frequency of only 11.6 - 34.9%) against the L. muta muta venom. The positive control (Doxorubicin) and the venom alone presented a damage frequency of 80.3% and 70.7%, respectively. The flower EO protected DNA from damage induced by L. muta muta venom. L. alba leaf and flower EOs presented anti-genotoxic action

Chemical composition, antimicrobial and antifungal activity of Lippia Thymoide essential oil in oral pathogens

Aim: This study evaluated the chemical composition of Lippia thymoides (Lt) essential oil and its antimicrobial activity against fungal strains of Candida albicans (Ca) and Gram-negative bacteria Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn). Methods: Lt essential oil was obtained by hydrodistillation apparatus with a modified Clevenger extension. The chemical analysis was analyzed by gas phase chromatography and mass spectrometry on Shimadzu QP 2010 plus. Sample sensitivity evaluation was performed by ABHb-inoculum and culture plates were developed with triphenyltetrazolium chloride, also Fn and Pi samples analysis were in anaerobic environment and Ca sample analysis was performed in aerobic environment. The minimum inhibitory concentration (CIM) was determinated by microdilution in eppendorfs tubes. Results: The chemical analysis showed that Thymol (59,91%) is the main compound found in Lt essential oil, also other antifungal and antimicrobial agents were present γ-terpinene (8.16%), p-cymene (7.29%) and ß-caryophyllene (4.49%), Thymol is a central ingredient of many medicinal plants and has a potent fungicidal, bactericidal and antioxidant activity, it has been previously shown to have anti-inflammatory activity against Periodontal Disease (PD) cause can reduces prostanoids, interleukins, leukotrienes levels in periodontium. CIM result Pi was 6.5 µg/mL, Fn was 1.5 µg/mL and Ca was 0.19 µg/mL. Conclusion: The antimicrobial activity of L. thymoides, through the compound Thymol, has been shown promising potential against gram-negative periodontopathogenic bacteria and fungi whose therapeutic arsenal is still very restricted

Morphometry of bovine blastocysts produced in vitro in culture media with antioxidants cysteamine or oily extract of Lippia origanoides / [Morfometria de blastocistos bovinos produzidos in vitro em meio de cultura com antioxidantes cisteamina ou extrato oleoso de Lippia origanoides]

This study aimed to evaluate the ultrastructural morphometry of bovine embryos produced in vitro grown at different concentrations of antioxidants. After in vitro maturation and fertilization, the presumptive zygotes were assigned into five treatments. T1) without the addition of any antioxidants (negative control); T2) addition of 50µM/mL cysteamine; and T3, T4 and T5) adding 2.5µg/mL, 5.0µg/mL or 10.0µg/mL of the antioxidants derived from the oily extract from Lippia origanoides, respectively. On D7 of culture, the embryos in the blastocyst stage were fixed and prepared for electron transmission microscopy. These were evaluated for the proportion of cytoplasm-to-nucleus, cytoplasm-to-mitochondria, cytoplasm-to-vacuoles, cytoplasm-to-autophagic vacuoles and cytoplasm-to-lipid droplets. Blastocysts cultured in media containing oily extract of Lippia origanoides presented morphological characteristics such as high cell:mitochondria ratio and low cell:vacuoles and cell:autophagic vacuole ratio, possibly been morphological indicators of embryonic quality. Inner cell mass (ICM) from blastocysts cultured in media without any antioxidants had the highest cell:vacuole ratio. Similar results were found in the trophectoderm (TE) cells of blastocysts from treatment 2. Embryo culture media supplemented with antioxidants derived from Lippia origanoides oil produced embryos with a higher cytoplasmic proportion of organelles, such as mitochondria. Also, treatments without any antioxidants or with the addition of cysteamine presented cytoplasmic vacuolization, a characteristic related to production of poor-quality embryos.(AU)

Este estudo teve como objetivo avaliar a morfometria ultraestrutural de embriões bovinos produzidos in vitro e cultivados em diferentes concentrações de antioxidantes. Após a maturação e a fertilização in vitro, os possíveis zigotos foram divididos em cinco tratamentos: T1) sem adição de antioxidantes (controle negativo); T2) adição de 50µM/mL de cisteamina; e T3, T4 e T5) adição de 2,5µg/mL, 5,0µg/mL ou 10,0µg/mL dos antioxidantes derivados do extrato oleoso de Lippia origanoides, respectivamente. No D7 de cultivo, os embriões em estágio de blastocisto foram fixados e preparados para microscopia eletrônica de transmissão. Estes foram avaliados para a proporção entre citoplasma e núcleo, citoplasma e mitocôndria, citoplasma e vacúolos, citoplasma e vacúolos autofágicos e citoplasma e gotículas lipídicas. Blastocistos cultivados em meio contendo extrato oleoso de Lippia origanoides apresentaram características morfológicas como alta relação célula:mitocôndria e baixa relação célula:vacúolos e célula:vacúolo autofágico, possíveis indicadores morfológicos de qualidade embrionária. A massa celular interna (MCI) de blastocistos cultivados em meio sem quaisquer antioxidantes teve a maior razão célula:vacúolo. Resultados semelhantes foram encontrados nas células do trofectoderma (TE) de blastocistos do tratamento 2. Portanto, o meio de cultivo embrionário suplementado com antioxidantes derivados do óleo de Lippia origanoides produziu embriões com maior proporção citoplasmática de organelas, como mitocôndrias. Além disso, tratamentos sem antioxidantes ou com adição de cisteamina apresentaram vacuolização citoplasmática, característica relacionada à produção de embriões de baixa qualidade.(AU)

Antibacterial activity of Lippia alba, Myrcia lundiana and Ocimum basilicum essential oils against six food-spoiling pathogenic microorganisms / Actividad antibacteriana de los aceites esenciales de Lippia alba, Myrcia lundiana y Ocimum basilicum contra seis microorganismos patógenos que estropean los alimentos

The aim of this study was to undertake a screening experiment on essential oils (EO) of Myrcia lundiana, Ocimum basilicum and Lippia alba against six food-spoiling pathogenic bacteria. Seventy-two (72) samples were initially analyzed fo antimicrobial activity based on the agar diffusion test. The minimum inhibitory (MIC) and bactericidal (MBC) concentrations were determined for the 12 samples which showed greatest antimicrobial potential in this stage. Two samples of L. alba, three samples of M. lundiana and seven samples of O. basilicum showed a MIC of 0.12-125 µL/mL for the six tested bacteria. Of these, the EO of O. basilicum cultivar Maria Bonita stood out with the lowest MIC and MBC. Thus, a mixture simulating this essential oil was prepared from commercial standards of the compounds (±)-linalool, geraniol and 1,8-cineole. Significantly higher MIC and MBC were detected in the simulation compared to the respective EO, suggesting a synergistic effect between compounds.

El objetivo de este estudio fue realizar un experimento de detección en aceites esenciales (AE) de Myrcia lundiana, Ocimum basilicum y Lippia alba contra seis bacterias patógenas que estropean los alimentos. Setenta y dos (72) muestras fueron analizadas inicialmente para la actividad antimicrobiana basada en la prueba de difusión en agar. Se determinaron las concentraciones mínimas inhibitoria (CMI) y bactericida (CMB) para las 12 muestras que mostraron el mayor potencial antimicrobiano en esta etapa. Dos muestras de L. alba, tres muestras de M. lundiana y siete muestras de O. basilicum mostraron un CMI de 0.12-125 µL/mL para las seis bacterias analizadas. De estos, el AE de O. basilicum cultivar Maria Bonita se destacó con el CMI y CMB más bajos. Por lo tanto, se preparó una mezcla que simula este aceite esencial a partir de los estándares comerciales de los compuestos de (±)-linalol, geraniol y 1,8-cineol. Se detectaron CMI y CMB significativamente más altos en la simulación en comparación con el AE respectivo, lo que sugiere un efecto sinérgico entre los compuestos.

Ação inseticida in vitro do óleo essencial de Lippia gracilis Schauer sobre Damalinia Bovicola ovis (Schrank, 1781) / In vitro action of Lippia gracilis Schauer essential oil on Damalinia Bovicola ovis (Schrank, 1781)

O objetivo deste trabalho é avaliar a eficácia inseticida in vitro do óleo essencial de Lippia gracilis sobre Bovicolaovis. Os espécimes (n = 900) de B.ovis foram coletados de ovinos naturalmente infestados, distribuídos aleatoriamente em grupos, e submetidos aos diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml óleo essencial de Lippia gracilis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). Realizou-se 4 réplicas (25 piolhos por réplica) para cada tratamento. A viabilidade dos piolhos foi avaliada nos tempos de 1, 3, 6, 24, 48 e 72 h. Os dados foram expressos em eficácia média ± desvio padrão (SPSS versão 23.0) e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo essencial de L. gracilis nas concentrações de 100 a 62,5 mg/mL apresentaram eficácia de 100%, 1 h pós-exposição. Os valores de CI50 e CI90 foram 18,1 mg/mL e 44,9 mg/mL, respectivamente. Dessa forma, concluímos que o óleo essencial de L. gracilis apresentou eficácia sobre B.ovis in vitro.

The objective of this work is to evaluate the in vitro insecticidal efficacy of the essential oil of Lippia gracilis against Bovicolaovis. The specimens (n = 900) of B.ovis were collected from naturally infested sheep, randomly distributed in groups, and subjected to different treatments: (G1) 1,000 to 15.6 mg/ml essential oil of Lippia gracilis; (G2) 3% Tween 80 (negative control), and: (G3) Amitraz 1.25 g/mL (positive control). 4 replicates (25 lice per replica) were performed for each treatment. Louse viability was assessed at 1, 3, 6, 24, 48 and 72 h. The data were expressed as mean efficacy ± standard deviation (SPSS version 23.0) and statistical differences in the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit regression was performed to obtain the IC50 and CI90, with a significance level of p <0.05. The essential oil of L. gracilis at concentrations of 100 to 62.5 mg/mL showed 100% efficacy, 1 h post-exposure. The IC50 and CI90 values were 18.1 mg/mL and 44.9 mg/mL, respectively. Thus, we conclude that L. gracilis essential oil is effective on B. ovis.

Ação inseticida in vitro do óleo essencial de Lippia gracilisSchauer sobre Damalinia Bovicola ovis (Schrank, 1781) / In vitro action of Lippia gracilis Schauer essential oil on Damalinia Bovicola ovis (Schrank, 1781)

O objetivo deste trabalho é avaliar a eficácia inseticida in vitro do óleo essencial de Lippia gracilis sobre Bovicolaovis. Os espécimes (n = 900) de B.ovis foram coletados de ovinos naturalmente infestados, distribuídos aleatoriamente em grupos, e submetidos aos diferentes tratamentos: (G1) 1.000 a 15,6 mg/ml óleo essencial de Lippia gracilis; (G2) Tween 80 a 3% (controle negativo), e: (G3) Amitraz 1,25 g/mL (controle positivo). Realizou-se 4 réplicas (25 piolhos por réplica) para cada tratamento. A viabilidade dos piolhos foi avaliada nos tempos de 1, 3, 6, 24, 48 e 72 h. Os dados foram expressos em eficácia média ± desvio padrão (SPSS versão 23.0) e diferenças estatísticas dos grupos experimentais foram obtidas por Kruskal-Wallis e Friedman. Para obtenção da CI50e CI90 foi realizada Regressão de Probit, com nível de significância de p< 0,05. O óleo essencial de L. gracilis nas concentrações de 100 a 62,5 mg/mL apresentaram eficácia de 100%, 1 h pós-exposição. Os valores de CI50 e CI90 foram 18,1 mg/mL e 44,9 mg/mL, respectivamente. Dessa forma, concluímos que o óleo essencial de L. gracilis apresentou eficácia sobre B.ovis in vitro.

The objective of this work is to evaluate the in vitro insecticidal efficacy of the essential oil of Lippia gracilis against Bovicolaovis. The specimens (n = 900) of B.ovis were collected from naturally infested sheep, randomly distributed in groups, and subjected to different treatments: (G1) 1,000 to 15.6 mg/ml essential oil of Lippia gracilis; (G2) 3% Tween 80 (negative control), and: (G3) Amitraz 1.25 g/mL (positive control). 4 replicates (25 lice per replica) were performed for each treatment. Louse viability was assessed at 1, 3, 6, 24, 48 and 72 h. The data were expressed as mean efficacy ± standard deviation (SPSS version 23.0) and statistical differences in the experimental groups were obtained by Kruskal-Wallis and Friedman. Probit regression was performed to obtain the IC50 and CI90, with a significance level of p <0.05. The essential oil of L. gracilis at concentrations of 100 to 62.5 mg/mL showed 100% efficacy, 1 h post-exposure. The IC50 and CI90 values were 18.1 mg/mL and 44.9 mg/mL, respectively. Thus, we conclude that L. gracilis essential oil is effective on B. ovis.

Study of the scientific production of the antibacterial activity of the chemical compounds of the essential oil of Lippia sidoides / Estudo da produção científica da atividade antibacteriana dos componentes químicos do óleo essencial de Lippia sidoides

Lippia sidoides Cham. has widespread use in folk medicine because its essential oil (EO) presents great antibacterial properties. In Brazil, this plant has been introduced in governmental programs for herbal medicine, due to its recognized therapeutic activities. This promoted greater interest in the search for new molecules with antimicrobial activity in this EO which have been described by several authors. Thus, this work aimed to present a study of the scientific production of antibacterial activity of the chemical compounds from L. sidoides essential oil (LSEO). The inclusion criteria were articles which assessed the chemical components and antibacterial activity through the inhibition diameter, Minimum Inhibitory Concentration (MIC), and Minimal Bactericidal Concentration (MBC), published between 2000 and 2020. Of the 996 studies identified, 55 met the inclusion criteria. In descending order, the most frequently detected chemical components found in LSEO were: thymol, p-cymene, and caryophillene. Regarding to the origin, the results showed that the extraction of EO were carried out mainly in the Northeast (55.5%) and Southeast (19%) regions of Brazil; and the part of the plant most used for the production were the leaves. In addition, the essential oil showed strong antibacterial activity against most of the bacteria tested.(AU)

Lippia sidoides Cham. é amplamente utilizada na medicina popular devido às propriedades antibacterianas atribuídas ao seu óleo essencial (OE). No Brasil, esta planta tem sido introduzida em programas governamentais de fitoterapia, por causa de suas atividades terapêuticas. Isso promoveu um maior interesse na busca por novas moléculas com atividade antimicrobiana presentes neste óleo, as quais foram relatadas por diversos autores. Assim, este trabalho objetivou apresentar um estudo da produção científica da atividade antibacteriana dos compostos químicos do OE da L. sidoide. Para tanto, foram utilizados artigos que abordaram os componentes químicos e atividade antibacteriana por meio de dados de diâmetro de inibição, Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM), publicados entre 2000 e 2020. De um total de 996 estudos identificados, 55 preencheram os critérios de inclusão. Em ordem decrescente, os componentes químicos mais comumente encontrados no OE da L. sidoide foram: timol, p-cimeno e cariofileno. Com relação à origem, os resultados mostraram que a extração do OE foi realizada principalmente nas regiões Nordeste (55,5%) e Sudeste (19%) do Brasil; e a parte da planta mais utilizada para a produção foram as folhas. Além disso, o OE apresentou uma forte atividade antibacteriana contra a maioria das bactérias testadas. (AU)

Efecto antinociceptivo y antiinflamatorio del aceite esencial de Lippia pedunculosa Hayek y su complejo de inclusión de ß-ciclodextrina / Antinociceptive and anti-inflammatory effect of Lippia pedunculosa Hayek essential oil and its ß-cyclodextrin inclusion complex

Lippia pedunculosa Hayek (EOLp) presenta efectos tripanocidas y amebicidas. En este trabajo se estudia su aceite esencial en modelos experimentales de analgesia e inflamación una vez que la prevalencia del dolor en la población genera un gran sufrimiento y discapacidad, y los medicamentos que se usan con mayor frecuencia tienen efectos secundarios indeseables. También se evalúa si la formulación del complejo de inclusión EOLp/ß-ciclodextrina (ß-CD) fue capaz de mejorar la actividad antinociceptiva de la EOLp sola. Los datos se evaluaron mediante análisis de varianza (ANOVA), seguido de la prueba de Tukey. Las diferencias se consideraron significativas si p<0,05. EOLp presentó un mejor efecto antinociceptivo en comparación con el complejo de inclusión EOLp/ß-CD. De esta manera, las ciclodextrinas parecen no ser eficientes para aceites esenciales con sustancias de peróxido. Sin embargo, en peritonitis, EOLp redujo la migración total de leucocitos y los niveles de IL-1ß en el líquido peritoneal, lo que confirma su efecto antiinflamatorio. Los efectos observados sugieren que EOLp es una buena y prometedora opción para el tratamiento de la inflamación y los trastornos relacionados con el dolor.

Lippia pedunculosa Hayek (EOLp) presents tripanocid and amebicid effects. However essential oil needs to be further studied in experimental models of analgesia and inflammation once the prevalence of pain in the population generates great suffering and disability and the drugs most often used have undesirable side effects. We also evaluated whether the inclusion complex formulation EOLp/ß-cyclodextrin (ß-CD) was able to improve the antinociceptive activity of the EOLp alone. Data were evaluated by analysis of variance (ANOVA), followed by Tukey's test. Differences were considered significant if p<0.05. EOLp presented better antinociceptive effect when compared to the EOLp/ß-CD inclusion complex. Thus, cyclodextrins appear not to be efficient for essential oils with peroxide substances. However, in peritonitis, EOLp reduced total leucocyte migration and IL-1ß levels in the peritoneal fluid, which confirmed its anti-inflammatory effect. The observed effects suggest that EOLp is the best promising option for the treatment of inflammation and pain-related disorders.

Larvicidal activity of Caesalpinia ferrea Mart. and Lippia origanoides Cham. Against Aedes aegypti (LINNAEUS, 1762) (Diptera: Culicidae)

The control of Aedes aegypti has been considered one of the most important public health challenges worldwide. Chemical compounds have long been used for this purpose, but resistance to these molecules has also increased. Therefore, over the last few years several studies have focused on the development of alternative tools, particularly those based on plant metabolites. The purpose of this study was to assess the larvicidal activity of Caesalpinia ferrea and Lippia origanoides against Ae. aegypti. Larvae (L3) of Ae. aegypti Liverpool and Rockefeller strains, as well as of the Recife population were exposed to different concentrations of C. ferrea (ranging from 13.1 to 105 mg/mL) and L. origanoides (ranging from 16.3 to 130 mg/mL), and the mortality rate was evaluated up to 48 hours after the beginning of the experiment. All tested groups and ontrol group were quadruplicated. For C. ferrea, mortality ranged from 42.5% to 100% for Ae. aegypti Liverpool strain, from 67% to 100% for Ae. aegypti Rockfeller strain, and 57% to 100% for Ae. aegypti Recife population after 48 hours of larval exposure. For L. origanoides, the larvicidal activity ranged from 75% to 100% for Ae. aegypti Liverpool strain, from 61.5% to 100% for Ae. aegypti Rockfeller strain, and from 60.5% to 100% for Ae. aegypti Recife population. The hydro ethanol extract of C. ferrea and L. origanoides presented larvicidal activity against Ae. aegypti.

In vitro anthelmintic activity of Lippia gracilis Schauer essential oil against egg-hatching of goat gastrointestinal nematodes / Atividade anti-helmíntica in vitro do óleo essencial de Lippia gracilis Schauer contra a eclosão de nematódeos gastrintestinais de caprinos

Since drug-resistant nematodes became a common problem in sheep and goat industries, alternative methods using natural products have emerged as a viable and sustainable anthelmintic treatment option. Here, the in vitro effect of essential oil extracted from Lippia gracilis Schauer was assessed on the hatching process of nematodes recovered from naturally infected goats. Essential oil at concentrations of 0.08% (0.008 µL/mL), 0.12% (0.012 µL/mL), and 0.16% (0.016 µL/mL) was able to induce an average inhibition of 74.7, 84 and 93%, respectively. The effective concentration required to inhibit egg hatching in 50% of eggs (EC50) was 0.03452%. Therefore, essential oil of L. gracilis showed promisor in vitro anthelmintic results against egg-hatching of goat gastrointestinal nematodes.(AU)

Como os nematoides resistentes a drogas se tornaram um problema comum nas indústrias de ovinos e caprinos, métodos alternativos que utilizam produtos naturais surgiram como uma opção de tratamento anti-helmíntico viável e sustentável. Aqui, o efeito in vitro do óleo essencial extraído de Lippia gracilis Schauer foi avaliado no processo de eclosão de nematoides recuperados de caprinos naturalmente infectadas. O óleo essencial nas concentrações de 0,08% (0,008 µL/mL), 0,12% (0,012 µL/mL), e 0,16% (0,016 µL/mL)foi capaz de induzir uma inibição média de 74,7, 84 e 93%, respectivamente. A concentração efetiva necessária para inibir a eclosão de ovos em 50% dos ovos (CE50) foi de 0,03452%. Portanto, o óleo essencial de L. gracilis apresentou resultados anti-helmínticos in vitro promissores contra a eclosão de nematódeos gastrintestinais de caprinos.(AU)

Content and chemical composition of the essential oil of Lippia gracilis Schauer accessions in different drying times / Teor e composição química do óleo essencial de acessos de Lippia gracilis Schauer em diferentes tempos de secagem

Lippia gracilis, popularly known in Brazil as 'alecrim-de-tabuleiro', is used for many purposes, especially as antimicrobial and antiseptic. The drying process of aromatic and medicinal plants aims to minimize the loss of active principles and slow their deterioration, which may greatly influence the yield and chemical composition of some species. The objective of this study was to evaluate the effect of drying times (0, 2, 4, and 8 days) on the content and chemical composition of the essential oil of L. gracilis accessions LGRA-106, LGRA-109, and LGRA-201. The leaves were dried at 40 oC, and essential oil was extracted by hydrodistillation. Chemical analysis was performed by GC/MS. The experiment was carried out in a completely randomized design with three replications. The accessions of L. gracilis LGRA-106, LGRA-109, and LGRA-201 presented higher essential oil at four days of drying time. The accession LGRA-201 showed the highest essential oil yields at four and eight days of drying, with mean values of 0.038 and 0.029 mL g-1, respectively. The drying time did not influence the contents of thymol, methyl-thymol, γ-terpinene, and carvacrol in the essential oils of L. gracilis, but affected the contents of ß-caryophyllene, p-cymene, and carvacrol acetate. The essential oils of the three accessions analyzed in this study revealed different chemical profiles.

Lippia gracilis, conhecida popularmente como alecrim-de-tabuleiro é usada para muitos efeitos, especialmente como antimicrobiano e antisséptico. O processo de secagem das plantas aromáticas e medicinais visa minimizar a perda de princípios ativos e retardar a sua deterioração os mesmos podem afetar sobremaneira o rendimento e a composição química de algumas plantas. O objetivo deste trabalho foiavaliar o efeito de diferentes tempos de secagem (0, 2, 4 e 8 dias) no teor e na composição química do óleo essencial dos acessos LGRA-106, LGRA-109 e LGRA-201 de L. gracilis.As folhas foram secas a 40 oC e a extração do óleo essencial foi por hidrodestilação. A análise química foi feita através CG/EM. O ensaio foi implantado em delineamento inteiramente casualizado com três repetições. Os acessos de L. gracilis LGRA-106, LGRA- 109 e LGRA-201 apresentaram maiores rendimentos de óleo essencial no tempo de secagem de quatro dias. O acesso LGRA-201 apresentou os maiores teores de óleo essencial aos quatro e oito dias de secagem, com valores médios de 0,038 e 0,029 mL g-1, respectivamente. O tempo de secagem não influenciou os teores de timol, metil-timol, terpineno e carvacrol nos óleos essenciais de L. gracilis, porém afetou os teores de ß-cariofileno, ρ-cimeno, and acetato de carvacrol. Os óleos essenciais dos três acessos analisados apresentaram perfis químicos diferentes entre si.