Two new nimbolinin- and trichilin-class limonoids isolated from the fruits of Melia azedarach / 中国天然药物

Two new furan fragment isomerized limonoids, meliazedalides A and B (compounds 1 and 2), were isolated from the fruits of Melia azedarach Linn.. Their chemical structures were elucidated on the basis of HR-ESI-MS and 1D and 2D NMR data, which belonged to nimbolinin- and trichilin-class, respectively. Compound 2 exhibited weak inhibitory effect on NO production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages with IC being 37.41 μmol·L.

Ovary histology and quantification of hemolymph proteins of Rhipicephalus (Boophilus) microplus treated with Melia azedarach / Histologia do ovário e quantificação de proteínas na hemolinfa de Rhipicephalus (Boophilus) microplus tratado com Melia azedarach

This study aimed to analyze ovary histology and quantify total protein in the hemolymph of Rhipicephalus (Boophilus) microplus females treated with hexane extracts from green fruits of Melia azedarach. Eight engorged females were immersed in the extract at 0.25% concentration, and eight in water containing 5% acetone (control). The females were dissected 72 hours after treatment, and the ovaries were weighed and subjected to standard histological techniques. The total protein concentration was measured in the hemolymph of 200 females, of which 100 were treated as described above and 100 served as a control. In the treated group, ovary weight reduction and predominance of immature oocytes were observed. In addition, there were decreases in the diameters of the cytoplasm and germ vesicle of the oocytes in the treated group, compared with the controls. The protein concentration in the hemolymph was higher in the treated group than in the controls. The morphological changes observed in the treated ovaries included: presence of vacuolization; alteration of oocyte morphology, which changed from rounded to elongated; deformation of the chorion; and disorganization of the yolk granules. These results demonstrate the action of M. azedarach fruit extracts on R. (B.) microplus oogenesis.

Este estudo foi desenvolvido, visando analisar a histologia do ovário e quantificar as proteínas totais na hemolinfa de fêmeas de Rhipicephalus (Boophilus) microplus tratadas com extrato hexânico de frutos verdes de Melia azedarach. Oito fêmeas ingurgitadas foram tratadas por imersão com o extrato na concentração de 0,25%, e oito com água contendo 5% de acetona (controle). As fêmeas foram dissecadas 72 horas após o tratamento e os ovários foram pesados e submetidos a técnicas histológicas padrões. A concentração total de proteína foi mensurada na hemolinfa de 200 fêmeas, sendo 100 tratadas como descrito anteriormente e 100 como controle. Foi observada redução do peso dos ovários, predomínio de ovócitos imaturos e houve diminuição nos diâmetros do citoplasma e da vesícula germinal dos ovócitos do grupo tratado em comparação ao controle. A concentração de proteína na hemolinfa foi mais alta no grupo tratado que no controle. As alterações morfológicas observadas nos ovários tratados foram a presença de vacuolizações, alteração da morfologia dos ovócitos que mudaram de redondos para alongados, deformação do córion e desorganização dos grânulos de vitelo. Estes resultados demonstram a ação do extrato de M. azedarach na ovogênese de R. (B.) microplus.

Atividade inseticida de extratos diclorometano e metanólico de Azadirachta indica (A. JUSS), Melia azedarach (L.) e Carapa guianenses (AUBL.) (Meliaceae) sobre cupim subterrâneo Coptotermes gestroi (WASMANN) (Isoptera, Rhinotermitidae) / Insecticidal activity of dichloromethane and methanolic extracts of Azadirachta indica (A. JUSS), Melia azedarach (L.) AND Carapa guianenses (AUBL.) (Meliaceae) on the subterranean termite Coptotermes gestroi (WASMANN) (Isoptera, Rhinotermitidae)

Cupins de hábito subterrâneo e os agroquímicos usados para controlá-los causam significativos prejuízos no meio ambiente urbano. Em virtude disso elaborou-se o presente trabalho, cujo o objetivo foi avaliar o efeito de extratos diclorometano e metanólico de três espécies da família Meliaceae, Azadirachta indica (A. JUSS), Melia azedarach (L.) e Carapa guianenses (AUBL.) sobre Coptotermes gestroi (WASMANN). Para tanto foram realizados dois experimentos em condições laboratoriais. O primeiro avaliou a sobrevivência dos insetos frente a condições ótimas (presença de umidade e alimento) e adversas (ausência de umidade ou alimento, ou de ambos os elementos). A umidade demonstrou ser o fator limitante, reduzindo de forma significativa a sobrevivência dos insetos (P < 0,05), seguida da ausência de alimento. O segundo experimento avaliou o efeito de extratos orgânicos das três plantas sobre C. gestroi. O experimento contou com seis tratamentos, e três grupos testemunhas (diclorometano, metanol e água). Os extratos que mais afetaram os insetos foram os de A. indica. AIDic que reduziu em 17 dias o tempo de sobrevivência, apresentando mortalidade superior a 70% a partir do sétimo dia, AIMet apresentou resultados semelhantes. Os extratos de M. azedarach (MADic) também reduziram significativamente a sobrevivência dos insetos (P < 0,05). Observou-se um claro efeito residual do solvente extrator, principalmente do metanol.

Subterranean termites and pesticides used to control them cause significant losses in the urban environment.Because of this the present work was elaborated; whose the objective was to evaluate the dichloromethane and methanol extracts effects of three species of the Meliaceae family, Azadirachta indica (A. JUSS), Melia azedarach (L.) and Carapa guianenses (AUBL.) on Coptotermes gestroi (WASMANN). For this purpose two experiments were carried out under laboratory conditions. The first one evaluated the survival of the insects front the excellent conditions (presence of humidity and food) and adverse (absence of humidity or food, or the both elements). The humidity demonstrated to be the limitant factor, by significant reduction of insects survival (P < 0,05), followed by the food lack. The second experiment evaluated the effect of organic extracts of three plants on C. gestroi. The experiment had six treatments, most controls: dichloromethane, methanol and water, with 10 repetitions each. The extracts that had more affected on the insects survival time was A. indica. AIDic (A. indica dichloro extract) reduced in 17 days the survival time, with 70% superior mortality from the seventh day, AIMet (A. indica methanolic extract) presented similar result. The dichloromethane extract of M. azedarach (MADic) also reduced significantly the insects survival (P < 0,05). A residual effect of the solvent extractor, mainly of methanol was observed clearly.

Actividad antibacteriana de extracto de hojas de Melia azedarach L / Antibacterial activity of leaf extract Melia azedarach L

El trabajo tuvo como objetivo evaluar la actividad antibacteriana de extractos de hojas de Melia azedarach L. sobre seis bacterias patógenas. Inicialmente mediante extracción por el método Soxhlet se obtuvo extracto total en etanol y a partir de éste se prepararon fracciones líquido-líquido con éter de petróleo y acetato de etilo. El extracto total y las dos fracciones fueron diluidos a diferentes concentraciones (ppm) para evaluar in vitro su actividad antibacteriana. Las bacterias de mayor susceptibilidad fueron las patógenas de humanos Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa y Klebsiella oxytoca, en relación a la fitopatógena Burkholderia glumae quien mostró resistencia a todos los tratamientos. Las bacterias patógenas fueron más susceptibles a la fracción éter de petróleo a concentración mínima de 25 ppm. El screen fitoquímico de la planta indicó presencia de metabolitos secundarios tipo alcaloides, terpenos/esteroles, saponinas, taninos y antocianinas. Estos resultados evidencian el posible uso de M. azederach como alternativa de control biológico sobre las bacterias analizadas.

The work was objective to evaluate the antibacterial activity of extracts from leaves of Melia azedarach (L) on six pathogenic bacteria. Total ethanol extract was obtained initially by extraction with method Soxhlet and from this prepared liquid-liquid fractions with petroleum ether and ethyl acetate. The total extract and the two fractions were diluted at different concentrations (ppm) to evaluate its antibacterial activity in vitro. More susceptible bacteria were the pathogenic human Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Klebsiella oxytoca, in relation to the plant pathogen Burkholderia glumae who showed resistance to all treatments. Pathogenic bacteria were more susceptible to the fraction of petroleum ether to the minimum concentration of 25 ppm. Screen plant phytochemical indicated presence of secondary metabolites type alkaloids, terpenes/sterols, saponins, tannins and anthocyanins. These results demonstrate the potential use of M. azederach as biological control alternatively on analysed bacterial.

The Toxicity and Anti-cancer Activity of the Hexane Layer of Melia azedarach L. var. japonica Makino's Bark Extract

In this study, the 4-week oral toxicity and anti-cancer activity of the hexane layer of Melia azedarach L. var. japonica Makino's bark extract were investigated. We carried out a hollow fiber (HF) assay and 28-day repeated toxicity study to confirm the anti-cancer effect and safety of the hexane layer. The HF assay was carried out using an A549 human adenocarcinoma cell via intraperitoneal (IP) site with or without cisplatin. In the result, the 200 mg/kg b.w of hexane layer with 4 mg/kg b.w of cisplatin treated group, showed the highest cytotoxicity aginst A549 carcinoma cells. For the 28-day repeated toxicity study, 6 groups of 10 male and female mice were given by gavage 200, 100, or 50 mg/kg b.w hexane layer with or without 4 mg/kg b.w of cisplatin against body weight, and were then sacrificed for blood and tissue sampling. The subacute oral toxicity study in mice with doses of 200, 100, and 50 mg/kg b.w hexane layer showed no significant changes in body weight gain and general behavior. The cisplatin-treated group significantly decreased in body weight compared to the control group but regained weight with 100 and 200 mg/kg b.w of hexane layer. The biochemical analysis showed significant increase in several parameters (ALT, total billirubin, AST, creatinine, and BUN) in cisplatin-treated groups. However, in the group given a co-treatment of hexane layer (200 mg/kg b.w), levels of these parameters decreased. In hematological analysis, cisplatin induced the reduction of WBCs and neutrophils but co-treatment with hexane layer (100 and 200 mg/kg b.w) improved these toxicities caused by cisplatin. The histological profile of the livers showed eosinophilic cell foci in central vein and portal triad in cisplatin treated mice. These results show that hexane layer might have an anti-cancer activity and could improve the toxicity of cisplatin.

Antibacterial efficacy of the seed extracts of Melia azedarach against some hospital isolated human pathogenic bacterial strains

<p><b>OBJECTIVE</b>To investigate the antibacterial potential of the polar and non-polar extracts of the seeds of Melia azedarach (M. azedarach) L. (Meliaceae) against eighteen hospital isolated human pathogenic bacterial strains.</p><p><b>METHODS</b>Petrol, benzene, ethyl acetate, methanol, and aqueous extracts at five different concentrations (1, 2, 5, 10 and 15 mg/mL) were evaluated. Disk diffusion method was followed to evaluate the antibacterial efficacy.</p><p><b>RESULTS</b>All extracts of the seeds demonstrated significant antibacterial activity against tested pathogens. Among all extracts, ethyl acetate extract revealed the highest inhibition comparatively. The present study also favored the traditional uses reported earlier.</p><p><b>CONCLUSIONS</b>Results of this study strongly confirm that the seed extracts of M. azedarach could be effective antibiotics, both in controlling gram-positive and gram-negative human pathogenic infections.</p>

Efeito de genótipos de tomateiro e de extratos aquosos de folhas de Melia azedarach e de sementes de Azadirachta indica sobre Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) / Effects of tomato genotypes and aqueous extracts of Melia azedarach leaves and Azadirachta indica seeds on Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

Insecticide plants are an important tool among the new alternatives for pest control in IPM systems because they reduce the use of synthetic insecticides, preserving human health and the environment. We investigated the effects of aqueous extracts of Melia azedarach leaves and Azadirachta indica seeds and three tomato genotypes, 'Santa Clara', 'IPA-5' - Solanum lycopersicum (= Lycopersicon esculentum Mill), and LA444-1 - S. peruvianum (= L. peruvianum), on the development, reproduction and longevity of the tomato pinworm Tuta absoluta (Meyrick), under laboratory conditions. The trials were set up in a completely randomized design, with nine treatments [three genotypes x two extracts (M. azedarach and A. indica) and control]. The replication consisted on five tubes, each with three newly hatched larvae, totalizing 90 individuals per treatment. The larvae were fed with tomato leaves treated with aqueous extracts at 0.1 percent concentration or distilled water (control) and daily observed until adults' emergence. Larval and pupal development and mortality, pupal weight, longevity and fecundity were evaluated. The accession LA444-1 negatively affected the development and reproduction of T. absoluta; the tomato pinworm had similar development and reproduction on 'IPA-5' and 'Santa Clara' (the susceptible control). The association of resistant tomato genotypes and extracts of M. azedarach leaves and neem seeds did not result in synergistic or antagonistic effects on T. absoluta.

Effect of morphological heterogeneity of somatic embryos of Melia azedarach on conversion into plants

Embryogenic cultures were initiated from immature Melia azedarach (Meliaceae) zigotic embryos. Explants were induced on Murashige and Skoog (1962) medium with 4.54 microM thidiazuron or 0.45 microM dichlorophenoxyacetic acid. After 6 weeks of culture on induction medium, somatic embryos were categorized in four morphological classes based on the presence of single or fused embryos and if they remained united or not to the original explant; that were evaluated histologically. The somatic embryos of every category were transferred, in groups or individually, on a 1/4 MS medium. Bipolar embryos, the more typically normal ones, had well defined shoot and root apical meristems and produced single plants; subcultured individually their conversion was 28%, and subcultured in groups the conversion declined to 6.8%. Fused embryos subcultured in groups had only a 2.1% conversion and produced plants with fused stems. None conversion rate in the others classes was associated to poorly developed shoot and root meristematic areas or with their absence. The converted plants were acclimatized and transferred, in a mist, to soil, with an independent of the class 95% survival rate.

Avaliação da eficácia de extratos oleosos de frutos verdes e maduros de cinamomo (Melia azedarach) sobre Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) / Evaluation of efficacy of ripe and unripe fruit oil extracts of Melia azedarach against Rhipicephalus (Boophilus) microplus (Acari: ixodidae)

Extratos hexânicos obtidos dos frutos de Melia azedarach foram testados sobre fêmeas ingurgitadas e larvas de Rhipicephalus (Boophilus) microplus. Foram produzidos quatro extratos: um de frutos verdes e três com frutos maduros, sendo um com os frutos imediatamente processados, outro com frutos armazenados por cinco meses e um testado após armazenamento do extrato por 2 (dois) anos a 4ºC. Os testes foram feitos em triplicata, em diluições de 0,25 por cento a 0,0156 por cento. A eficácia dos extratos foi avaliada através da comparação da Reprodução Estimada dos lotes "controle e tratado" para fêmeas, da mortalidade para as larvas, e do cálculo das DL50 e da potência relativa para larvas e fêmeas. O extrato de frutos verdes apresentou uma eficácia variando de 3,6 por cento a 100 por cento para fêmeas, e o de frutos maduros variou de 5,2 por cento a 99,7 por cento. Os dois extratos causaram uma mortalidade de 100 por cento das larvas nas maiores concentrações. O extrato de frutos armazenados apresentou eficácia de 14 por cento a 83 por cento sobre fêmeas, enquanto o extrato armazenado em geladeira apresentou eficácia de 8,4 por cento a 100 por cento e ambos não apresentaram efeito larvicida. Os resultados obtidos demonstram uma superioridade do extrato verde, com uma menor DL50 e potência 1,497 vezes superior ao extrato de frutos maduros.

Hexanic extracts gotten of the fruits of cinamomo (Melia azedarach) were tested on engourged females and larvae of Rhipicephalus (Boophilus) microplus. Four extracts were produced: one of unripe fruits and three of ripe fruits, being one with the fruits immediately processed, another one with fruits stored for five months and one tested after storage of the extract for 2 (two) years at 4ºC. The tests were made in triplicate, in 0.25 percent- 0.0156 percent dilutions. The effectiveness of extracts was evaluated through the comparison of the Reproduction Estimate of the groups "controlled and treated" for females, of mortality for the larvae, and of the calculation of the DL50 and the relative power for larvae and females. The extract of unripe fruits presented the effectiveness varying the 3.6 percent - 100 percent for females, and of ripe fruits it varied of 5.2 percent - 99.7 percent. The two extracts caused a mortality of 100 percent of the larvae in the highest concentrations. The extract of stored fruits presented effectiveness of 14 percent - 83 percent on females, while the extract stored in refrigerator presented effectiveness of 8.4 percent 100 percent and both did not present larvicidal effect. The results demonstrate a superiority of the unripe extract, with a minor DL50 and superior power 1.497 times to the extract of ripe fruits.

Antifertility effect of Melia azedarach Linn. (dharek) seed extract in female albino rats.

In the present study, the effect of oral administration of Melia azedarach Linn. (dharek) seed extract on fertility index, uterine weight and various histological and biochemical parameters of uterus were studied in the adult cyclic Wistar rats. Average number of embryos and implantation losses in the pregnant animals treated with dharek seed extract was also studied. The extract was prepared using a flash evaporator at 35 degrees C and dissolved in olive oil to prepare doses on per kg body weight basis. The results indicated a reduction in fertility index and average number of embryos in mated rats treated with the dharek extract. Pre-implantation, post-implantation and total prenatal mortalities were increased in rats treated with dharek seed extract during early (D1-D7) and late (D7-D18) stages of gestation period at doses of 5, 10 and 20 mg kg(-1) body wt day(-1). Histological studies showed a significant reduction in myometrial thickness, uterine gland diameter, luminal diameter of uterine glands and luminal epithelial cell height in rats treated with dharek seed extract at 1mg kg(-1) body wt day(-1) for 18 days. Pits and folds in luminal epithelial, mitotic activity in luminal and glandular epithelial cells of uterus were observed to be absent. Biochemically, a significant increase in protein and glycogen contents was observed. Thus, in conclusion, the application of this plant extract in rodent control programme may help to elevate the socioeconomic status of the society.

Intoxicação experimental com frutos de Melia azedarach (Meliaceae) em suínos / Experimental poisoning by fruits of Melia azedarach (Meliaceae) in pigs

Frutos maduros, triturados, de Melia azedarach foram administrados misturados à ração a 8 suínos em doses únicas de 5-20g/kg. Diarréia passageira foi o único sinal nos 2 suínos que consumiram 5g/kg. Os sinais clínicos observados nos animais que ingeriram doses únicas de 10, 15 e 20g/kg caracterizaram-se por incoordenação, tremores musculares, dificuldade para manter-se de pé, relutância para levantar-se, decúbito esternal e hipotermia. Os 2 suínos que ingeriram 20g/kg morreram. Macroscopicamente observou-se congestão do intestino delgado e da mucosa do estômago. As alterações histológicas caracterizaram-se por discreta a moderada necrose do tecido linfático, necrose dos músculos esqueléticos, congestão e necrose da mucosa gástrica e do intestino delgado e alterações degenerativas moderadas no fígado e rim. Os níveis séricos de AST e CPK aumentaram após a ingestão do tóxico. Observaram-se valores altos de CPK nos animais que morreram, evidenciando a mionecrose. Os resultados obtidos demonstram que, para suínos, a ingestão de altas doses de frutos de Melia azedarach induz ao aparecimento de sinais clínicos da intoxicação e causa lesões em diversos órgãos, inclusive a morte.

Extracts of Azadirachta indica and Melia azedarach seeds inhibit folliculogenesis in albino rats

The seed oil of Azadirachta indica A. Juss (neem) is used in traditional medicine for its antidiabetic, spermicidal, antifertility, antibacterial, and wound healing properties. The present study was undertaken to investigate the quantitative aspects of follicular development in cyclic female albino rats (135 ± 10 g; 8 groups with 6 animals in each group) after oral administration of polar (PF) and non-polar (NPF) fractions of A. indica seed extract at 3 and 6 mg kg body weight-1 day-1 and Melia azedarach Linn. (dharek) seed extract at 24 mg kg body weight-1 day-1 for 18 days. The extracts were prepared using a flash evaporator at 35°C and then dissolved in olive oil to prepare doses on a per kg body weight basis. There was a significant reduction (P = 0.05) in the number of normal single layered follicles (A. indica: 0.67 ± 0.33 and 4.67 ± 2.03 after 3 and 6 mg/kg NPF, and 3.33 ± 1.67 and 1.00 ± 1.00 after 3 and 6 mg/kg PF vs control: 72.67 ± 9.14 and M. azedarach: 0.60 ± 0.40 and 1.80 ± 1.2 after 24 mg/kg PF and NPF, respectively, vs control: 73.40 ± 7.02) and follicles in various stages (I-VII) of follicular development in all treatment groups. These extracts also significantly reduced (P = 0.05) the total number of normal follicles in the neem (14.67 ± 5.93 and 1.00 ± 1.00 after 3 and 6 mg/kg PF and 3.67 ± 0.88 and 5.33 ± 2.03 after 3 and 6 mg/kg NPF) and dharek (13.00 ± 3.58 and 14.60 ± 2.25 after 24 mg/kg NPF and PF) treatments compared to control (216.00 ± 15.72 and 222.20 ± 19.52, respectively). Currently, indiscriminate use of persistent and toxic rodenticides to control rodent populations has created serious problems such as resistance and environmental contamination. Therefore, it becomes necessary to use ecologically safe and biologically active botanical substances that are metabolized and are not passed on to the next trophic level, and that interfere with the reproductive potential particularly growth and differentiation of follicles. This may help elevate the socio-economic status of the country. Thus, the present study is an attempt to investigate the effects of A. indica and M. azedarach seed extracts on reproduction of albino rats.

Ação do extrato hexânico de frutos maduros de Melia azedarach (Meliaceae) sobre Boophilus microplus (Acari: Ixodidae) em bezerros infestados artificialmente / Efficacy of Melia azedarach (Meliaceae) hexanic ripe fruit extract against Boophilus microplus (Acari: Ixodidae) on artificially infested calves

A eficácia de um extrato hexânico dos frutos de Melia azedarach foi avaliada em bezerros infestados artificialmente com Boophilus microplus. Os frutos foram submetidos ao processo de secagem e, depois de moídos, o extrato foi produzido em Soxhlet utilizando-se o hexano como extrator. Oito bezerros infestados artificialmente, cinco vezes a cada cinco dias com 2.500 larvas de B. microplus, foram divididos em dois grupos (tratado e controle) com base na infestação por carrapatos. No dia zero os animais foram banhados com 3 litros de uma solução a 0,25por cento do extrato em água e acetona (1,2 litro). Os animais do grupo controle foram banhados com o mesmo volume de água e acetona. Diariamente, as fêmeas ingurgitadas desprendidas naturalmente eram contadas e 40 eram incubadas para acompanhamento dos parâmetros reprodutivos. O número médio de fêmeas, o índice de conversão em ovos (peso da massa de ovos/ peso do grupo de fêmeas x 100) e o percentual de eclosão, obtidos nos dois grupos, foram comparados mediante o teste T de Student. Após 21 dias de contagens, o número médio de fêmeas ingurgitadas foi significativamente menor (P<0,05) no grupo tratado (188) do que no controle (247). Porém, a conversão e a eclodibilidade das larvas não foram afetadas pelo tratamento. Estes resultados indicam que houve interferência da planta no desenvolvimento do carrapato sobre os animais, mas não na sua reprodução. Comparados com a literatura, os resultados se revelaram promissores. No entanto, novos estudos nos quais se empregue uma formulação adequada em uma concentração mais elevada são necessários para se propor o uso desta planta no controle deste carrapato.

Influence of genotype and explant source on indirect organogenesis by in vitro culture of leaves of Melia azedarach L

In vitro regeneration of shoots from leaf explants of the Paradise tree (Melia azedarach L.) was studied. Three different portions (proximal portion, distal portion and rachis of the leaflets) of three developmental stages (folded, young still expanding and completely expanded) of leaves of 10-15 year old plants of seven genotypes were cultured on Murashige and Skoog (1962) medium (MS) supplemented with 1 mg x l(-1) benzylaminopurine (BAP) + 0.1 mg x l(-1) kinetin (KIN) + 3 mg x l(-1) adenine sulphate (ADS). The rachis of the leaflets of the completely expanded leaves was found to be the most responsive tissue, in most of the genotypes employed. Shoot regeneration occurred in leaf explants of all the genotypes tested. The best genotype for shoot regeneration was clone 4. Rooting was induced on MS medium supplemented with 2.5 mg x l(-1) 3-indolebutyric acid, IBA, (4 days) followed by subculture on MS lacking growth regulators (26 days). Complete plants were transferred to soil.