Tagetes erecta L.: A traditional medicine effective in inflammatory process treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Tagetes erecta L. (Asteraceae), popularly known as Aztec Marigold, is used in folk medicine to treat several ailments including inflammatory processes. Despite its historical use, the specific mechanisms through which it may modulate inflammation, particularly its effects on neutrophils and macrophages activation, have not yet been completely investigated. AIM OF THE STUDY: This study aimed to elucidate the anti-inflammatory mechanism of the hydroalcoholic extract obtained from T. erecta flowers, focusing on its role in the regulation of neutrophil and macrophage functions. MATERIAL AND METHODS: The production of TNF, IL-6, CXCL-1, IL-1ß, IL-10 (ELISA) and NO (Griess reaction), adhesion molecule expression (CD62L, CD49d and CD18, flow cytometry), and chemotaxis were analyzed in vitro using oyster glycogen-recruited peritoneal neutrophils or macrophages (RAW 264.7) stimulated with lipopolysaccharide (LPS) and treated with the extract (1, 10 or 100 µg/mL). The resolution of inflammation was accessed by efferocytosis assay. The in vivo anti-inflammatory activity was investigated using carrageenan-induced inflammation in the subcutaneous tissue of male Swiss mice orally treated with the T. erecta extract (30, 100 or 300 mg/kg). The leukocyte influx (optical microscopy), secretion of chemical mediators (TNF, IL-6 and IL-1ß, ELISA) and protein exudation (Bradford reaction) were quantified in the inflamed exudate. RESULTS: In vitro studies demonstrated that the extract inhibited neutrophil chemotaxis and reduced the production and/or release of cytokines (TNF, IL-1ß, CXCL1, and IL-6) as well as nitric oxide (NO) by neutrophils and macrophages when stimulated with LPS. Neutrophils treated with LPS and incubated with the extract showed an increase in CD62L expression, which leads to the impairment of neutrophil adhesion. The extract also enhanced efferocytosis of apoptotic neutrophils by macrophages, which was accompanied by increased IL-10 secretion and decreased TNF levels. In vivo studies yielded similar results, showing reduction in neutrophil migration, protein exudation, and cytokine release (TNF, IL-6, and IL-1ß). CONCLUSIONS: Together, the data herein obtained shows that T. erecta flower extract has anti-inflammatory effects by regulating inflammatory mediators, limiting neutrophil migration, and promoting efferocytosis. The in vivo results suggest that an herbal medicine made with T. erecta could represent an interesting pharmacological tool for the treatment of acute inflammatory condition.

Isolation of flavonoids from Tagetes patula (French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells / Aislación de flavonoides de las flores de Tagetes patula (cempasúchil francés): Actividad citotóxica y oxidante en células de carcinoma cervical humano

T agetes patula , known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessed the cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patula flowers as anti - cancer cervical agents. Fraction F6 of n - butanol extract was subjected to column chromatography and HPLC - ESI - MS. The isolated compo unds of T. patula were used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patula have great treatment p otential against cervical cancer.

Tagetes patula , conocida como cempasúchil francés, pertenece a la familia Asteraceae. La infección por el virus del papiloma humano se considera una de las causas del cáncer cervical. En este estudio, se evaluó la actividad citotóxica y la capacidad oxidativa intracelular de los compuestos aislados del extracto de las flores de T. patula como agentes anticancerígenos cervicales. La fracción F6 del ext racto de n - butanol se sometió a cromatografía en columna y HPLC - ESI - MS. Los compuestos aislados de T. patula se utilizaron para examinar la actividad citotóxica y la producción total de especies reactivas de oxígeno en las células SiHa y HeLa; las células también se caracterizaron mediante microscopía electrónica de barrido. Patulitrina resultó citotóxica para las células SiHa y HeLa. Se observó un aumento en la producción de ROS en diferentes momentos del tratamiento de las células con patuletina y patulit rina. La microscopía electrónica de barrido mostró cambios morfológicos en las células SiHa y HeLa. Por lo tanto, los compuestos aislados de T. patula tienen un gran potencial de tratamiento contra el cáncer cervical.

Effect of Tagetes minuta essential oil on the central nervous system of unfed Rhipicephalus sanguineus sensu lato 'tropical lineage' ticks.

Rhipicephalus sanguineus sensu lato 'tropical lineage' (Acari: Ixodidae) is considered a sanitary concern due to its role as a disease vector. Tick strains resistant to synthetic acaricides have caused difficulties in their control, besides synthetic acaricides are harmful to the environment and to the health of non-target animals. The research of plants with acaricidal and repellent properties has proved to be an efficient alternative in tick control. The genus Tagetes spp. excels for its use as traditional pest control in households and plantations and also for its potential as an acaricide against R. sanguineus under laboratory conditions. The first aim of the present study was to evaluate the effect of different doses of Tagetes minuta essential oil (TMEO) on the central nervous system (synganglion) in unfed R. sanguineus adults. The histological analysis of synganglion exposed to the different concentrations of TMEO and amitraz 12.5% (50% of the recommended dose in the package insert) showed a significant effect with signs of cell damage including volume increase, loss of shape, and vacuolization, in addition to chromatin alterations such as condensation, margination, and fragmentation. TMEO were analyzed by gas chromatography coupled with mass spectrometry showing the presence of 21 compounds that according to their chemical structure are classified as terpenoids. Among them (Z)-ß-ocimene, ocimene, (Z)-tagetone, and verbenone were found in major quantities.

Cytotoxic and Antitumor Effects of the Hydroalcoholic Extract of Tagetes erecta in Lung Cancer Cells.

Among all cancers, lung cancer is the one with the highest mortality rate, and it also has limited therapeutics. Antitumor agents based on medicinal plants have gained importance as a source of bioactive substances. Tagetes erecta is a plant of great cultural value, and recent reports have suggested its cytotoxic effects in tumor cells. Our objective was to evaluate the antitumor activity of Tagetes erecta extract in a lung carcinoma model. Hydroalcoholic extracts were obtained from fresh flowers and leaves of T. erecta; both extracts did not exert toxicity on Artemia salina. We observed cytotoxic effects induced by the floral extract in Lewis lung carcinoma (LLC) and breast tumor cell line (MCF7), but not by the leaf extract. In vivo, a xenograft lung carcinoma model was performed with LLC cells implanted on C57BL/6 mice, which showed that the floral extract reduced tumor growth and improved the effect of etoposide. Microscopic analysis of tumors showed a reduction in mitoses and an increase in necrotic areas with the extract and the etoposide. The main phytochemical compounds found are 2,3-dihydro-benzofuran, octadecanoic acid, benzenacetic acid, oleic acid, linoleic acid, and acetic acid. We conclude that the hydroalcoholic extract of T. erecta flowers has cytotoxic effects in lung carcinoma cells and enhances the effect of etoposide.

Extract of Tagetes erecta L., a medicinal plant rich in lutein, promotes gastric healing and reduces ulcer recurrence in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Tagetes erecta L. (Asteraceae), popularly known as Aztec Marigold, is used in South America to treat several ailments. Despite reports that T. erecta flowers are used in folk medicine to treat gastrointestinal diseases, there is no study regarding its gastric healing effects. AIM OF THE STUDY: The effect of dry extract of T. erecta L. (DETe) in gastric healing and gastric ulcer recurrence was evaluated, contributing to the validation of the antiulcer potential of this medicinal plant. METHODS: Rats were treated orally with vehicle (1 ml/kg), omeprazole (20 mg/kg), or DETe (3, 30 or 300 mg/kg) for 7 days, twice a day. The lesion area was evaluated, and the levels of reduced glutathione (GSH) and lipoperoxides (LOOH) and the activity of the superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and myeloperoxidase (MPO) were measured. The ulcer recurrence was evaluated in mice and induced by interleukin (IL)-1ß (1 µg/kg, i.p). The recurred area, gastric wall thickness, GSH and cytokines levels, MPO and N-acetylglucosaminidase (NAG) activities were measured. RESULTS: DETe accelerated the healing of gastric ulcers only at 300 mg/kg, reducing the ulcerated area by 66%. In parallel, DETe reduced LOOH levels, SOD, CAT and MPO activities, while increasing GST activity and mucin amount. In the recurrence model, DETe reduced the lesion area by 94%, and in parallel decreased the gastric wall thickness and TNF levels, while increasing IL-10 amount. CONCLUSIONS: Corroborating the popular use of T. erecta, DETe favors the antioxidant system and reduce gastric inflammation, accelerating the gastric healing process and reducing the ulcer recurrence.

Tagetes erecta L. flowers, a medicinal plant traditionally used to promote diuresis, induced diuretic and natriuretic effects in normotensive and hypertensive rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Tagetes erecta L., known as marigold, belongs to the Asteraceae family and is mainly found in South America. Despite reports that T. erecta flowers are used in folk medicine to treat cardiovascular and renal diseases, there is no study regarding its diuretic effect. AIM: This study aimed to evaluate the chemical composition and the diuretic efficacy of the hydroethanolic extract from T. erecta (HETE) in normotensive (NTR) and hypertensive (SHR) rats. MATERIAL AND METHODS: The HETE was analyzed by liquid chromatography coupled to diode array detector and mass spectrometry (LC-DAD-MS). Female and male NTR and SHR received the treatment with vehicle, HETE (0.01 mg/kg, 0.1 mg/kg, and 1 mg/kg) or hydrochlorothiazide (HCTZ; 5 mg/kg) orally. The urinary parameters were measured at the end of the 8-h experiment. RESULTS: From HETE, saccharides and triterpenes were the main annotated compounds, such as erythrodiol and ß-amyrin. The urine volume was significantly increased in the groups treated with HETE, in both male and female NTR and SHR rats, compared to the respective vehicle-treated groups. Regarding electrolytes elimination, the treatment with HETE did not reveal significant changes in the urine levels of K+ or Cl-, but it showed a natriuretic and Ca2+-sparing effects. The HETE beneficial result in reducing Ca2+ excretion was confirmed through the protective effect found in front of the urinary calcium oxalate precipitation and crystallization. The combination with HCTZ, a classic diuretic and saluretic medicine, significantly enhanced HETE-induced diuresis, natriuresis, and the Ca2+-sparing effect. On the other hand, the K+-sparing action was improved in the combination of HETE with amiloride, a standard K+-sparing diuretic. In contrast, the combination of HETE with atropine (a non-selective muscarinic receptor antagonist) and indomethacin (an inhibitor of the cyclooxygenase enzyme), promoted an important reduction in urinary volume, but interestingly the natriuretic effect was maintained. CONCLUSION: This study contributed to the preclinical validation of the diuretic efficacy of T. erecta, highlighting this species as promising for the development of new pharmacological strategies for the management of kidney disorders.

Antimicrobial and physical properties of experimental endodontic sealers containing vegetable extracts.

To assess the antimicrobial activity and the physical properties of resin-based experimental endodontic sealers with the incorporation of vegetable extracts obtained from Bixa orellana, Mentha piperita, and Tagetes minuta species. The extracts were obtained and characterized by gas chromatography-mass spectrometry (GC-MS), and minimum inhibitory concentration (MIC) against Streptococcus mutans, Enterococcus faecalis, and Candida albicans. The extracts were individually incorporated into a dual-cure experimental sealer at a mass concentration of 0.5%. A commercial reference RealSeal was used. The sealers were evaluated by measuring the setting time, degree of conversion, dimensional stability, radiopacity, flow, and film thickness of these materials, also and its antimicrobial effect was evaluated using the direct contact test. Data were statistically analyzed by analysis of variance and Tukey's post-hoc test at α = 0.05 significance level. The physical properties were not influenced by the addition of the vegetable extracts (p > 0.05). For S. mutans, only T. minuta and B. orellana groups presented antibacterial activity after 24 h of contact (p < 0.05). All extracts evidenced an antibacterial effect against E. faecalis (p < 0.05). The experimental sealers hold promise as a novel vegetable sealer with great antimicrobial activity and also great physical-mechanical properties. Nonetheless, more studies are needed.

Sunlight Photocatalytic Performance of ZnO Nanoparticles Synthesized by Green Chemistry Using Different Botanical Extracts and Zinc Acetate as a Precursor.

In this work, the assessment of Azadirachta indica, Tagetes erecta, Chrysanthemum morifolium, and Lentinula edodes extracts as catalysts for the green synthesis of zinc oxide nanoparticles (ZnO NPs) was performed. The photocatalytic properties of ZnO NPs were investigated by the photodegradation of methylene blue (MB) dye under sunlight irradiation. UV-visible (UV-Vis) spectroscopy, Fourier Transform Infrared (FTIR) spectroscopy, Transmission Electron Microscopy (TEM), X-ray Diffraction (XRD), Thermogravimetric (TGA), and Brunauer-Emmett-Teller analysis (BET) were used for the characterization of samples. The XRD results indicate that all synthesized nanoparticles have a hexagonal wurtzite crystalline structure, which was confirmed by TEM. Further, TEM analysis proved the formation of spherical and hemispherical nanoparticles of ZnO with a size in the range of 14-32 nm, which were found in aggregate shape; such a size was well below the size of the particles synthesized with no extract (~43 nm). ZnO NPs produced with Tagetes erecta and Lentinula edodes showed the best photocatalytic activity, matching with the maximum adsorbed MB molecules (45.41 and 58.73%, respectively). MB was completely degraded in 45 min using Tagetes erecta and 120 min using Lentinula edodes when subjected to solar irradiation.

Role of ß-Caryophyllene in the Antinociceptive and Anti-Inflammatory Effects of Tagetes lucida Cav. Essential Oil.

Tagetes lucida Cav. (Asteraceae) is an ancient medicinal plant commonly used to alleviate pain. Nevertheless, scientific studies validating this property are lacking in the literature. Animal models of pain were used to evaluate the antinociceptive and anti-inflammatory activities of T. lucida essential oil (TLEO) and a bioactive metabolite. The chemical constitution and possible toxicity of the extract and the mechanism of action of ß-caryophyllene were also explored. Temporal course curves and dose-response graphics were generated using TLEO (0.1-10 mg/kg or 3.16-31.62 mg/kg) and ß-caryophyllene (3.16-10 mg/kg). Metamizole (80 mg/kg) and indomethacin (20 mg/kg) were used as reference drugs in the formalin assay and writhing test in rats and mice, respectively. The ß-caryophyllene mechanism of action was explored in the presence of naloxone (1 mg/kg), flumazenil (10 mg/kg), WAY100635 (0.16 mg/kg), or nitro-l-arginine methyl ester (L-NAME) (20 mg/kg) in the formalin test in rats. GC/MS analysis demonstrated the presence of geranyl acetate (49.89%), geraniol (7.92%), and ß-caryophyllene (6.27%). Significant and dose-dependent antinociceptive response was produced by TLEO and ß-caryophyllene without the presence of gastric damage. In conclusion, ß-caryophyllene was confirmed as a bioactive compound in the T. lucida analgesic properties by involving the participation of receptors like opioids, benzodiazepines, and Serotonin 1A receptor (5-HT1A), as well as nitric oxide.

Photodynamic activity of Tagetes minuta extracts against superficial fungal infections.

Candida and dermatophyte species are the most common causes of superficial mycoses because their treatment can be difficult due to limitations of current antifungal drugs in terms of toxicity, bioavailability, interactions, narrow-spectrum activity, and development of resistance. Photodynamic therapy (PDT) involves the topical administration of a photosensitizer in combination with light of an appropriate wavelength and molecular oxygen that produces reactive oxygen species (ROS), which promote damage to several vital components of the microorganism. Tagetes species are known as a source of thiophenes, biologically active compounds whose antifungal activity is enhanced by irradiation with UVA. The present investigation evaluated Tagetes minuta extracts as a photosensitizer on growth of Candida and dermatophytes and their effect on Candida virulence factors. T. minuta root hexane and dichloromethane extracts demonstrated high photodynamic antifungal activity. Bioautographic assays and chromatographic analysis revealed the presence of five thiophenes with reported photodynamic antifungal activities under UVA. Analysis of ROS production indicated that both type I and II reactions were involved in the activity of the extracts. In addition, the extracts inhibited virulence factors of Candida, such as adherence to epithelial surfaces and germ tube formation and showed efficacy against different Candida morphologies: budding cells, cells with germ tube and biofilms. Results suggested that PDT with T. minuta extracts might become a valuable alternative to the already established antifungal drugs for the treatment of superficial fungal infections.

Chemical composition and antifungal activity of essential oils from Senecio nutans, Senecio viridis, Tagetes terniflora and Aloysia gratissima against toxigenic Aspergillus and Fusarium species.

Essential oils from aerial parts of Senecio nutans, Senecio viridis, Tagetes terniflora and Aloysia gratissima were analysed by GC-MS and their antifungal activities were assayed on toxigenic Fusarium and Aspergillus species. Sabinene (27.6 ± 0.1%), α-phellandrene (15.7 ± 0.3%), o-cymene (9.6 ± 0.2%) and ß-pinene (6.1 ± 0.2%) in S. nutans, 9,10-dehydrofukinone (92.7 ± 0.2%) in S. viridis, ß-thujone (36.1 ± 0.1%), α-thujone (32.2 ± 0.2%), 1,8-cineol (10.7 ± 0.1%) and sabinene (6.2 ± 0.2%) in A. gratissima, and cis-tagetone (33.6 ± 0.2%), cis-ß-ocimene (17.1 ± 0.2%), trans-tagetone (17.0 ± 0.1%), cis-ocimenone (8.0 ± 0.2%) and trans-ocimenone (8.2 ± 0.1%) in T. terniflora. The oils showed moderate antifungal activity (1.2 mg/mL > MIC >0.6 mg/mL) on the Fusarium species and a weak effect on Aspergillus species. The antifungal activity was associated on F. verticillioides to the high content of cis-tagetone, trans-tagetone, cis-ß-ocimene, cis-ocimenone, trans-ocimenone and on F. graminearum due to the total content of oxygenated sesquiterpenes and 9,10- dehydrofukinone. The oil of S. viridis synergized the effect of fungicides and food preservatives on F. verticillioides.[Formula: see text].

Compuestos fenólicos de Tagetes lucida Cav. con efecto antibacteriano por daños a la membrana / Phenolic compounds of Tagetes lucida Cav. with antibacterial effect due to membrane damage

Tagetes lucida Cav. (Asteraceae = Compositae) se usa para tratar infecciones estomacales. El estudio se centró en evaluar la composición y el efecto antimicrobiano de un extracto de T. lucida Cav. La planta se extrajo con etanol al 10% p/v, y la composición del extracto se analizó por Rp-HPLC-MS. El efecto antibacteriano se evaluó contra Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa y Salmonella choleraesuis utilizando métodos de difusión por disco, microdilución y bioautografía. Los ensayos de sytox y cometa fueron utilizados para evaluar el mecanismo de acción. De esta forma, se detectaron nueve compuestos fenólicos en el extracto de T. lucida. El extracto exhibió actividad solo en S. aureus (MIC de 4.000 mg/ml). La bioautografía reveló que los compuestos fenólicos podrían actuar sinérgicamente. Las pruebas de sytox y cometa mostraron una acción antibacteriana del extracto sobre la membrana bacteriana y el ADN de esta cepa bacteriana.

Tagetes lucida Cav. (Asteraceae=Compositae) is used for treating stomach infections. The study focused on evaluating the composition and antimicrobial effect of an extract of T. lucida Cav. The plant extracted with ethanol at 10% w/v, and the extract composition analyzed by Rp-HPLC-MS. The antibacterial effect was evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Salmonella choleraesuis using disk diffusion, microdilution and bioautography methods. The sytox and comet assays were used to evaluate the mechanism of action. In this way, nine phenolic compounds were detected in the extract of T. lucida. The extract exhibited activity only on S. aureus (MIC of 4.000 mg/ml). The bioautography revealed that the phenolic compounds could act synergistically. The sytox and comet tests showed an antibacterial action of the extract on the bacterial membrane and DNA of this bacterial strain.

Evaluation of Tagetes patula (Asteraceae) as an ecological alternative in the search for natural control of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The cattle tick Rhipicephalus microplus is a great sanitary problem and causes huge losses to livestock, being a vector of important diseases. The aim of this work was to verify the action of plant-derived material obtained from Tagetes patula on the life cycle of R. microplus, as well as to analyze the action of these extracts on ovary cells of engorged females. In the Adult Immersion Test, the crude ethanolic extract (TpEtOH) showed an efficacy of 99.2% (LC50 = 18.60 mg mL-1). From the repellency test, it was found that both the TpEtOH and essential oil were 100% efficient on the larval stage. In addition, in the Larval Package Test, a mortality of 98.37% was obtained with TpEtOH (LC50 = 3.798 mg mL-1). From the microscopic analysis of ovary cells, morphological changes in the chorion and cytoplasm of oocytes were observed, with vacuolization around the germinal vesicle and disorganization of the pedicular cells, suggesting an interference in the normal embryogenic formation of the larvae. These results demonstrate that T. patula extracts interfere at all stages of development of the ixodid, from eggs to adults forms, and have a pronounced repellent effect. In addition, the results of the cytotoxicity assays performed on keratinocytes, as well as previous information on oral and dermal acute toxicity (LD50 > 4000 mg kg-1), attest that T. patula can be a safe alternative for ectoparasitic control, representing an alternative for development of a formulation used to help control populations of ticks in the field.

Chemical Composition and In vitro Anthelmintic Activity of Extracts of Tagetes patula Against a Multidrug-Resistant Isolate of Haemonchus contortus.

Sheep breeding has suffered economic losses due to parasitism by gastrointestinal nematodes, particularly Haemonchus contortus. The use of natural products, specifically Tagetes patula, has been suggested as an alternative method of combatting this issue. Chemical analyses of the extracts of this species described in the literature report the presence of important classes of secondary metabolites such as thiophenes, flavonoids, alkaloids, and benzofurans, some of which were identified and isolated in this study. The aim of this work was to test the effect of the essential oil (EO) and the ethanolic extract of the aerial parts (TpEtOH ) of T. patula on eggs and larvae of H. contortus, through an egg hatch test (EHT) and a larval development test (LDT). In the EHT, the EO showed 100% inhibition at 0.75 mg mL-1 (LC50 = 0.0780 mg mL-1 ), and the TpEtOH showed 100% inhibition at 100 mg mL-1 (LC50 = 12.8 mg mL-1 ). In the LDT, the EO showed 100% inhibition at 0.375 mg mL-1 (LC50 = 0.0400 mg mL-1 ), and the TpEtOH showed 100% inhibition at 1.56 mg mL-1 (LC50 = 0.340 mg mL-1 ). Compared to available literature data, the results presented here suggest that the crude extracts of T. patula have substantial potential for controlling this nematode by interrupting its life cycle and/or preventing it from reaching the infective stage.

Extracts and semi-purified fractions of Tagetes patula flowers in the control of root-knot nematodes / Extratos e frações semipurificadas de flores de Tagetes patula no controle do nematoide-das-galhas

The genus Tagetes constitutes a group of antagonistic plant species that are cytotoxic against plant pathogenic nematodes, with T. patula being particularly efficient. The aim of this study was to evaluate the in vitro effect of extracts and semi-purified fractions of T. patula flowers on eggs and second-stage juveniles (J2) of Meloidogyne incognita, M. javanica, and M. paranaensis, as well as to verify the nematicidal effect of the flavonoids present in T. patula flowers. Extracts and semi-purified fractions were obtained from dried T. patula flowers after maceration and liquidification, yielding crude aqueous (CAE) and crude ethanol-water (CEWE) extracts. Dried flowers were also treated with n-hexane to obtain a crude defatted aqueous extract (CDAE) and a crude ethanol-water defatted extract (CEWDE). Then, the CEWE was fractionated, and the ethyl acetate (EAF), ethanol (EF), methanol (MF), and ethanol:water (EWF) fractions were obtained. CAE, CEWE, CDAE, and CEWDE were tested to evaluate their effects on hatching, mobility, and mortality of J2 of M. incognita, M. javanica, and M. paranaensis. EAF, EF, MF, and EWF fractions were tested on the same variables of M. incognita. All extracts significantly reduced J2 hatching of M. incognita, M. javanica, and M. paranaensis when compared to water and water + DMSO. CEWE had nematicidal effects on the three evaluated species, whereas CEWDE demonstrated nematicidal effects against M. incognita and M. javanica, and nematostatic effects on M. paranaensis. This toxic effect showed by CEWE may be related to the high content of quercetin, a major substance present in this sample. It was also observed that EAF accentuated the nematicidal response on Meloidogyne spp., suggesting that other medium polarity (methoxylated) flavonoids act as nematotoxic substances. Thus, these results suggest that quercetin contributes significantly to the nematicidal activity of CEWE and EAF.

Entre as espécies de plantas antagonistas, as do gênero Tagetes apresentam citotoxicidade contra fitonematoides, sendo que T. patula mostra-se eficiente. Assim, objetivou-se avaliar o efeito in vitro de extratos e frações semipurificadas de flores de T. patula sobre ovos e juvenis de segundo estádio (J2) de Meloidogyne incognita, M. javanica e M. paranaensis, e a capacidade nematicida dos flavonoides presentes nas flores de T. patula. Os extratos e frações semipurificadas foram preparados a partir de flores secas de T. patula, que após maceração foram submetidas à turbólise, obtendo-se os extratos bruto aquoso (EBA) e bruto etanol-água (EBEA). Também houve tratamento de flores secas com n-hexano para a obtenção do extrato bruto aquoso desengordurado (EBAD) e do extrato bruto etanol-água desengordurado (EBEAD). Em seguida, o EBEA foi fracionado, sendo obtidas as frações acetato de etila (FAE), etanólica (FE), metanólica (FM) e etanol:água (FEA). O efeito dos extratos EBA, EBEA, EBAD e EBEAD foram avaliados sobre a eclosão, mobilidade e mortalidade de J2 de M. incognita, M. javanica e M. paranaensis. As frações FAE, FE, FM e FEA foram avaliadas frente a M. incognita usando as mesmas variáveis. Os extratos reduziram significativamente a taxa de eclosão de J2 de M. incognita, M. javanica e M. paranaensis, quando comparados com água e água + DMSO. Quanto ao efeito dos extratos, observou-se que EBEA apresentou efeito nematicida para as três espécies avaliadas, e EBEAD demonstrou capacidade nematicida contra M. incognita e M. javanica, e sendo nematostático para M. paranaensis. Tal capacidade pode estar relacionada com o alto teor de quercetina (substância majoritária no EBEA). Observou-se, também, que o fracionamento químico e a obtenção da FAE acentuaram a resposta nematicida sobre Meloidogyne spp., sugerindo que outros flavonoides de média polaridade (metoxilados) atuam como substâncias nematotóxicas.

Extracts and semi-purified fractions of Tagetes patula flowers in the control of root-knot nematodes / Extratos e frações semipurificadas de flores de Tagetes patula no controle do nematoide-das-galhas

The genus Tagetes constitutes a group of antagonistic plant species that are cytotoxic against plant pathogenic nematodes, with T. patula being particularly efficient. The aim of this study was to evaluate the in vitro effect of extracts and semi-purified fractions of T. patula flowers on eggs and second-stage juveniles (J2) of Meloidogyne incognita, M. javanica, and M. paranaensis, as well as to verify the nematicidal effect of the flavonoids present in T. patula flowers. Extracts and semi-purified fractions were obtained from dried T. patula flowers after maceration and liquidification, yielding crude aqueous (CAE) and crude ethanol-water (CEWE) extracts. Dried flowers were also treated with n-hexane to obtain a crude defatted aqueous extract (CDAE) and a crude ethanol-water defatted extract (CEWDE). Then, the CEWE was fractionated, and the ethyl acetate (EAF), ethanol (EF), methanol (MF), and ethanol:water (EWF) fractions were obtained. CAE, CEWE, CDAE, and CEWDE were tested to evaluate their effects on hatching, mobility, and mortality of J2 of M. incognita, M. javanica, and M. paranaensis. EAF, EF, MF, and EWF fractions were tested on the same variables of M. incognita. All extracts significantly reduced J2 hatching of M. incognita, M. javanica, and M. paranaensis when compared to water and water + DMSO. CEWE had nematicidal effects on the three evaluated species, whereas CEWDE demonstrated nematicidal effects against M. incognita and M. javanica, and nematostatic effects on M. paranaensis. This toxic effect showed by CEWE may be related to the high content of quercetin, a major substance present in this sample. It was also observed that EAF accentuated the nematicidal response on Meloidogyne spp., suggesting that other medium polarity (methoxylated) flavonoids act as nematotoxic substances. Thus, these results suggest that quercetin contributes significantly to the nematicidal activity of CEWE and EAF.(AU)

Entre as espécies de plantas antagonistas, as do gênero Tagetes apresentam citotoxicidade contra fitonematoides, sendo que T. patula mostra-se eficiente. Assim, objetivou-se avaliar o efeito in vitro de extratos e frações semipurificadas de flores de T. patula sobre ovos e juvenis de segundo estádio (J2) de Meloidogyne incognita, M. javanica e M. paranaensis, e a capacidade nematicida dos flavonoides presentes nas flores de T. patula. Os extratos e frações semipurificadas foram preparados a partir de flores secas de T. patula, que após maceração foram submetidas à turbólise, obtendo-se os extratos bruto aquoso (EBA) e bruto etanol-água (EBEA). Também houve tratamento de flores secas com n-hexano para a obtenção do extrato bruto aquoso desengordurado (EBAD) e do extrato bruto etanol-água desengordurado (EBEAD). Em seguida, o EBEA foi fracionado, sendo obtidas as frações acetato de etila (FAE), etanólica (FE), metanólica (FM) e etanol:água (FEA). O efeito dos extratos EBA, EBEA, EBAD e EBEAD foram avaliados sobre a eclosão, mobilidade e mortalidade de J2 de M. incognita, M. javanica e M. paranaensis. As frações FAE, FE, FM e FEA foram avaliadas frente a M. incognita usando as mesmas variáveis. Os extratos reduziram significativamente a taxa de eclosão de J2 de M. incognita, M. javanica e M. paranaensis, quando comparados com água e água + DMSO. Quanto ao efeito dos extratos, observou-se que EBEA apresentou efeito nematicida para as três espécies avaliadas, e EBEAD demonstrou capacidade nematicida contra M. incognita e M. javanica, e sendo nematostático para M. paranaensis. Tal capacidade pode estar relacionada com o alto teor de quercetina (substância majoritária no EBEA). Observou-se, também, que o fracionamento químico e a obtenção da FAE acentuaram a resposta nematicida sobre Meloidogyne spp., sugerindo que outros flavonoides de média polaridade (metoxilados) atuam como substâncias nematotóxicas.(AU)

In vitro assessment of Argemone mexicana, Taraxacum officinale, Ruta chalepensis and Tagetes filifolia against Haemonchus contortus nematode eggs and infective (L3) larvae.

Argemone mexicana, Taraxacum officinale, Ruta chalepensis and Tagetes filifolia are plants with deworming potential. The purpose of this study was to evaluate methanolic extracts of aerial parts of these plants against Haemonchus contortus eggs and infective larvae (L3) and identify compounds responsible for the anthelmintic activity. In vitro probes were performed to identify the anthelmintic activity of plant extracts: egg hatching inhibition (EHI) and larvae mortality. Open column Chromatography was used to bio-guided fractionation of the extract, which shows the best anthelmintic effect. The lethal concentration to inhibit 50% of H. contortus egg hatching or larvae mortality (LC50) was calculated using a Probit analysis. Bio-guided procedure led to the recognition of an active fraction (TF11) mainly composed by 1) quercetagitrin, 2) methyl chlorogenate and chlorogenic acid. Quercetagitrin (1) and methyl chlorogenate (2) did not show an important EHI activity (3-14%) (p < 0.05); however, chlorogenic acid (3) showed 100% of EHI (LC50 248 µg/mL) (p < 0.05). Chlorogenic acid is responsible of the ovicidal activity and it seems that, this compound is reported for the first time with anthelmintic activity against a parasite of importance in sheep industry.

Anthelmintic effect of Psidium guajava and Tagetes erecta on wild-type and Levamisole-resistant Caenorhabditis elegans strains.

ETHNOPHARMACOLOGICAL RELEVANCE: Psidium guajava and Tagetes erecta have been used traditionally to treat gastrointestinal parasites, but their active metabolites and mechanisms of action remain largely unknown. AIM OF THE STUDY: To evaluate the anthelmintic potential of Psidium guajava and Tagetes erecta extracts on Levamisole-sensitive and Levamisole-resistant strains of the model nematode Caenorhabditis elegans. MATERIALS AND METHODS: Aqueous extracts of Psidium guajava (PGE) and Tagetes erecta (TEE) were assayed on locomotion and egg-laying behaviors of the wild-type (N2) and Levamisole-resistant (CB193) strains of Caenorhabditis elegans. RESULTS: Both extracts paralyzed wild-type and Levamisole-resistant nematodes in a dose-dependent manner. In wild-type worms, TEE 25mg/mL induced a 75% paralysis after 8h of treatment and PGE 25mg/mL induced a 100% paralysis after 4h of treatment. PGE exerted a similar paralyzing effect on N2 wild-type and CB193 Levamisole-resistant worms, while TEE only partially paralyzed CB193 worms. TEE 25mg/mL decreased N2 egg-laying by 65% with respect to the untreated control, while PGE did it by 40%. CONCLUSIONS: Psidium guajava leaves and Tagetes erecta flower-heads possess hydrosoluble compounds that block the motility of Caenorhabditis elegans by a mechanism different to that of the anthelmintic drug Levamisole. Effects are also observable on oviposition, which was diminished in the wild-type worms. The strong anthelmintic effects in crude extracts of these plants warrants future work to identify their active compounds and to elucidate their molecular mechanisms of action.

Insecticidal activity of an essential oil of Tagetes patula L. (Asteraceae) on common bed bug Cimex lectularius L. and molecular docking of major compounds at the catalytic site of ClAChE1.

Emerging resistance to insecticides has influenced pharmaceutical research and the search for alternatives to control the common bed bug Cimex lectularius. In this sense, natural products can play a major role. Tagetes patula, popularly known as dwarf marigold, is a plant native to North America with biocide potential. The aim of this work was to evaluate the biological activity of T. patula essential oil (EO) against adult common bed bugs via exposure to dry residues by the Impregnated Paper Disk Test (IPDT) using cypermethrin as a positive control. We selected the enzyme acetylcholinesterase as a target for modeling studies, with the intent of investigating the molecular basis of any biological activity of the EO. Chemical analysis of the EO was performed using gas chromatography coupled to mass spectrometry (GC-MS). Additionally, oral and dermal acute toxicity tests were performed according to Organization for Economic Cooperation and Development (OECD) guidelines. The sulforhodamine B assay (SRB) was performed to verify the cytotoxicity of EO to HaCaT cells. The EO eliminated 100 % of the bed bugs at 100 mg mL-1 with an LC50 value of 15.85 mg mL-1. GC-MS analysis identified α-terpinolene, limonene, piperitenone, and piperitone as major components of the mixture. Molecular modeling studies of these major compounds suggested that they are acetylcholinesterase inhibitors with good steric and electronic complementarity. The in vitro cytotoxicity evaluation revealed a LC50 = 37.06 µg mL-1 and in vivo acute toxicity showed an LC50 >4000 mg kg-1, indicating that the EO presents low risk of toxic side effects in humans. The T. patula essential oil components provide a promising strategy for controlling bed bug populations with low mammalian toxicity. These findings pave the way for further in vivo studies aimed at developing a safe and effective insecticide.

Nematicidal Activity of the Essential Oil of Three Varieties of Tagetes minuta from Argentina.

Essential oils composition of three Tagetes minuta varieties and a wild population (WP) from Argentina and their in vitro and in vivo nematicidal activity against root-knot nematode, Meloidogynejavanica, are described. All T. minuta EOs tested were very active against nematode juveniles (J2), but the strongest nematicidal effects were exhibited by the TmV3 variety oil, characterized by a high content of (E)-ocimenone. High nematode egg hatching suppression (> 90%) was induced by TmV3 EO after five days of incubation. In vivo tests on tomato seedlings showed a significant reduction of infection rate of M javanica J2 treated with TmV3 and WP oils at sub lethal dose. Therefore, EOs from chemically stable new varieties of T. minuta could be environmentally friendly nematicidal agents.