The effects of neem [Azadirachta indica A. Juss (meliaceae)] oil on Fusarium oxysporum f. sp. medicagenis and Fusarium subglutinans and the production of fusaric acid toxin

Fungi of the genus Fusarium are well known plant pathogens, cause several vascular diseases and are producers of toxins. In vitro assays evaluated the effects of Neem (Azadirachta indica) oil on the diameter of colonies, dry weight, spore production, spore viability and production of Fusaric Acid toxin on Fusarium oxysporum f. sp. medicagenis and Fusarium subglutinans isolates. Effects of Neem oil were analyzed at concentrations 0.25%, 0.5% and 1% in Czapek Yeast Agar medium. The production of Fu- saric acid was determined by Thin Layer Chromatography and quantified by UV spectrophotometry. Neem oil showed inhibitory effects on the isolates tested, although they varied according to type of isolate and oil concentration. Neem oil was efficient in reducing the colonies’ diameter and dry weight in concentration-dependent manner. Neem oil was efficacious at higher concentration in the decrease of sporulation. Spore germination was affected by Neem oil when the spore was grown in Neem-contained medium as when the spore emerged from a culture in a Neem medium. Neem oil decreased and even inhibited the production of Fusaric acid by the assayed isolates. Since these isolates are plant pathogens and producers of Fusaric acid, Neem oil may be introduced as an integral item in the management of host plants.

In vitro activity of neem oil [Azadirachta indica A. Juss (Meliaceae)] on Aspergillus flavus growth, sporulation, viability of spores, morphology and Aflatoxins B1 and B2 production

The effectiveness of neem (Azadiracta indica) oil on the growth, morphology, sporulation, viability of spores, aflatoxin B1 and B2 production by A. flavus on Yeast Extract-Sucrose medium was determined. Neem oil inhibited the fungal growth (i.e. mycelia dry weight, diameter of colony and growth rate) on solid media at concentrations from 0.5 to 5.0% v/v, although it significantly increased sporulation in the same conditions. Spores obtained from cultures grown without neem oil reduced germination when incubated in a neem oil supplemented medium. Colonies grown on solid media and in submerged cultures in the presence of neem oil exhibited morphological alterations, including granular cytoplasm, atypical hyphae branching pattern, abnormal and undifferentiated conidiophores. High Performance Liquid Chromatography was used to measure aflatoxins. In submerged cultures, neem oil at concentrations from 0.5 to 4.0% v/v caused approximately 95% inhibition in Aflatoxin B1 and B2. On other hand, these conditions failed to suppress fungal growth. Current research emphasized that neem oil was not fungistatic or fungicidal, but exhibited anti-aflatoxigenic activity.

Chemical constituents and biological activities of Azadirachta indica A. Juss

Azadirachta indica A. Juss is a tropical evergreen plant, belonging to the Meliaceae family. The plant is used as folk medicines for the treatment of fever, neuromuscular pains, skin infections, diabetes and heart disease. Previous researches show that the plant possesses several types of chemical constituents and diverse biological activities. The paper concludes chemical constituents, biological activities and pesticide products of A. indica.

Synthesis of plant-based biogenic jarosite nanoparticles using Azadirachta indica and Eucalyptus gunni leaf extracts and its application in Fenton degradation of dicamba

Bio-jarosite,an iron mineral synthesized biologically using bacteria,is a substitute for iron catalysts in the Fenton oxidation of organic pollutants.Iron nanocatalysts have been widely used as Fenton catalysts because they have a larger surface area than ordinary catalysts,are highly recyclable,and can be treated efficiently.This study aimed to explore the catalytic properties of bio-jarosite iron nanoparticles syn-thesized with green methods using two distinct plant species:Azadirachta indica and Eucalyptus gunni.The focus was on the degradation of dicamba via Fenton oxidation.The synthesized nanoparticles exhibited different particle size,shape,surface area,and chemical composition characteristics.Both particles were effective in removing dicamba,with removal efficiencies of 96.8%for A.indica bio-jarosite iron nano-particles(ABFeNPs)and 93.0%for E.gunni bio-jarosite iron nanoparticles(EBFeNPs)within 120 min of treatment.Increasing the catalyst dosage by 0.1 g/L resulted in 7.6%and 43.0%increases in the dicamba removal efficiency for EBFeNPs and ABFeNPs with rate constants of 0.025 min^(-1) and 0.023 min^(-1),respectively,confrming their catalytic roles.Additionally,the high efficiency of both catalysts was demonstrated through five consecutive cycles of linear pseudo-first-order Fenton oxidation reactions.

Molluscicidal effect of green synthesized silver nanoparticles using Azadirachta indica on Biomphalaria alexandrina snails and Schistosoma mansoni cercariae

Objective:To assess the molluscicidal effect of the eco-friendly green synthesized neem silver nanoparticles(neem-Ag NPs)against Biomphalaria alexandrina,the snail intermediate host for Schistosoma mansoni,and their cercaricidal potential.Methods:Methanol extracts from neem fruits were used for green synthesis of neem-Ag NPs.The neem-Ag NPs were characterized using UV-visible absorption spectra,dynamic laser light scattering technique,and transmission electron microscopy.The potential molluscicidal effect against adult and juvenile Biomphalaria alexandrina and the effect of the sub-lethal concentration on hatching of snail eggs and Schistosoma mansoni cercariae were evaluated.Results:The surface plasmon resonance of neem-Ag NPs showed a sharp absorption peak atλ_(max)=518 nm together with multiple peaks.The hydrodynamic diameter was(77.15±34.53)nm,the polydispersity index(0.338±0.000)and the zeta-potential-14.07 mV.Moreover,transmission electron microscopy showed that the average size of the nanoparticles was(27±2)nm.Agglomeration was evident and a light-colored capping layer could be seen coating the nanoparticles.Juvenile snails(LC_(50):0.83 ppm)were more susceptible to neem-Ag NPs than adults(LC_(50):1.07 ppm).In addition,neem-Ag NPs and neem at LC_(50)concentrations inhibited the egg-hatching of snails and showed cercaricidal activity in a time-dependent manner.Conclusions:Neem-Ag NPs have lethal activities against Biomphalaria alexandrina snails and their eggs,as well as Schistosoma mansoni cercariae.Hence,neem-Ag NPs could be a potential agent to control schistosomiasis.

A new cytotoxic salannin-class limonoid alkaloid from seeds of Azadirachta indica A. Juss

A new cytotoxic salannin-class limonoid alkaloid, azadiramide A（1）,was isolated from seeds of Azadirachta indica A. Juss. Its structure was elucidated by extensive analysis of spectroscopic data and quantum chemical calculation. Compound 1 is a rare salannin-class limonoid alkaloid. To our best knowledge, only two compounds belonging to this type were so far found. It showed inhibitory activity against human breast cancer MDA-MB-231 cell line with IC_（50） value of 2.70±0.63 μmol/L.

印楝Azadirachta Indica A.Juss的冷驯化与抗冻蛋白的研究

采用木本植物材料——印楝,通过组织培养建立快繁体系,然后对其进行冷驯化处理,并分析检测印楝植物体内抗冻蛋白.主要结果如下:①冷驯化处理后印楝的总蛋白一些表现为量的增加同时会有新的蛋白产生.但脱驯化或处理时间过长时,抗冻蛋白在量的表达上会有逐渐减少或消失的现象.②在对印楝的冷驯化中,发现不同的温度处理后蛋白稳定存在的时间不同.抗冻蛋白出现的最早时期为5℃处理2周左右,印楝能耐受的稳定最低温为5℃,所持续的最长时间约为20d.在0℃低温处理后,虽然在处理初期(0～15d)也有抗冻蛋白的产生,但随处理时间的延长,这种差异逐渐减少,在处理30d时完全消失.③得到了分离纯化的抗冻蛋白,其相对分子质量约为3.6×104.

印楝属(Azadirachta A.Juss.)植物分类及分布的研究现状

印楝属(Azadirachta A.Juss.)属楝科(Meliaceae)。印楝属植物由印度-马来亚(Indo-Malesian)特有成分组成,目前,世界分布(天然分布和栽培分布)约80个国家。现在得到公认并广泛使用的印楝属,传统上被归入楝属(Melia L.),由于与楝科的Antelaea、Azedarach和Trichilia的亲缘关系相近、形态特征相似,在属的归属上和种的确定上有较大的争议,被正式描述的"属"和"种"分别为5属、11种。从传统分类学水平,分析回顾了印楝属的系统位置、分类的现状与问题。并根据其地理分布,综述了印楝属植物的原产地及当今世界分布情况。

印楝(Azadirachta indica A.)高频植株再生系统的建立

[目的]建立印楝高频植株再生系统。[方法]以印楝带芽茎段为外植体,以MS为基本培养基,分别附加6-BA0.1、0.3、0.5、0.8、1.0mg/L和NAA0、0.1、0.3、0.5mg/L组配14个培养基配方处理进行芽分化的诱导培养试验,生根培养采用MS+IBA0.2mg/L、1/2MS和MS的3种培养基,研究外植体最适宜的消毒时间和取材部位,筛选诱导印楝芽分化和生根的最佳培养基配方。[结果]初代培养中,印楝外植体最适宜的消毒时间为10～12min,最适宜的取材部位为茎尖及植株的上部。14个培养基处理中,诱导芽分化的最佳培养基为:MS+6-BA0.5mg/L+NAA0.1mg/L+0.65%琼脂+3%蔗糖(pH5.8)。诱导生根的最佳培养基为:1/2MS+0.65%琼脂+3%蔗糖(pH5.8),生根率85.13%,小苗移栽成活率80%。[结论]该研究为印楝规模化快繁和遗传转化体系的建立奠定了基础。

不同林龄杂交楝(Azadirachta indica×Melia azedarach)人工林土壤水土保持功能研究

为了明确林龄对杂交楝(印楝×苦楝)人工林土壤水土保持功能的影响,通过测定土壤持水能力、渗透能力和抗蚀能力来综合评价不同林龄杂交楝人工林保持水土功能。结果表明,随着杂交楝林龄增加,林地土壤孔隙结构得到改善、持水性能增加、渗透能力增大,提高了土壤水稳性指数,增加了土壤抗蚀性,土壤水土保持功能得到增强。3年生杂交楝林分的土壤持水性能、渗透能力和抗蚀性与1年生林分之间无显著差异,5年生杂交楝林地各指标均比1年生和3年生林分有显著提高。5年生杂交楝林地的土壤最大持水量、总孔隙度、非毛管孔隙度、平均渗透速率和水稳性指数分别比1年生林地增加18.6%、11.4%、19.8%、45%、30.1%。因此,种植杂交楝可以在短期内提高林分的土壤水土保持功能,在山地丘陵地区值得大力发展杂交楝。

Neem(Azadirachta indica): Prehistory to contemporary medicinal uses to humankind

The divine tree neem(Azadirachta indica)is mainly cultivated in the Indian subcontinent.Neemhas been used extensively by humankind to treat various ailments before the availability ofwritten records which recorded the beginning of history.The world health organization estimatesthat 80%of the population living in the developing countries relies exclusively on traditionalmedicine for their primary health care.More than half of the world’s population still relies entirely on plants for medicines,and plants supply the active ingredients of most traditionalmedical products.The review shows the neem has been used by humankind to treat variousailments from prehistory to contemporary.

Toxicity profile of ethanolic extract of Azadirachta indica stem bark in male Wistar rats

Objective:To investigate the loxic implications of ethanolic stem bark extract of Azadirachta indica(A-indica)at 50,100,200 and 300 mg/kg body weight in Wistar rats.Methods:Fifty male rats of Wistar strains were randomly grouped into five(A-F.)of ten animals each.Animals in Group A(control)were orally administered 1 mL of distilled water on daily basis for 21 days while those in Groups B-E received same volume of the extract corresponding to 50,100,200 and 300mg/kg body weight.Results:The extract did not significantly(P>0.05)alter the levels of albumin,total protein,red blood cells and factors relating to it whereas the white blood cell,platelets,serum triacylglycerol and high-density lipoprotein cholesterol decreased significantly(P<0.05).In contrast,the final body weights,absolute weights of the liver,kidney,lungs and heart as well as their organ-body weight ratios,serum globulins,total and conjugated bilirubin,serum cholesterol,low-density lipoprotein cholesterol and computed atherogenic index increased significantly.The spleen-body weight ratio,alkaline phosphatase,alanine and aspartate transaminases,sodium,potassium,calcium,feed and water intake were altered at specific doses.Conclusions:Overall,the alterations in the biochemical parameters of toxicity have consequential effects on the normal functioning of the organs of the animals.Therefore,the ethanolic extract of A.indica stem bark at the doses of 50,100,200 and 300 mg/kg body weight may not be completely safe as an oral remedy and should be taken with caution if absolutely necessary.

Variability in permeability and integrity of cell membrane and depletion of food reserves in neem(Azadirachta indica) seeds from trees of different age classes

We quantified cell membrane permeability （electrical conduc-tivity-EC, water soluble sugar-WSS, and amino acids-AA） and integrity （phospholipids,α-tocopherol and lipid peroxidation） along with food reserve deterioration （total proteins, total sugar, and total starch） of neem seeds collected from various mother tree age classes and stored for 65 days in airtight plastic containers at ambient room temperature （35±5°C）. Results show that the activities were higher in fresh seeds （EC 267.56-2950.01 μS/g, WSS 19.96-19.48 mg/g and AA 5.40-5.35 mg/g） and declined with increasing duration of storage period （EC 153.37-195.17 μS/g, WSS 3.13-4.17 mg/g and AA 4.29-4.49 mg/g after 35 days and EC 144.02-161.56 μS/g, WSS 2.06-2.40 mg/g and AA 3.98-4.27 mg/g after 65 days of storage）. Phospholipids andα-tocopherol were higher in fresh seed （0.073-0.093 OD at 710 nm and 0.080-0.105 OD, respectively） and declined as storage duration in-creased （0.033-0.042 OD at 710 nm and 00.0010-0.0020 OD, respec-tively）. Dead seeds showed reduced amounts of phospholipids and minimum activity ofα- tocopherol （antioxidants）. The level of MDA was lower in fresh seeds （0.0066-0.0087 OD at 600-535 nm） and increased as storage duration increased （0.0248-0.0268 OD after 65 days of stor-age）. The higher amount of MDA indicated that seeds died due to rancid-ity of the oil inside the seed. Neem seed cake was assessed for deteriora-tion of food reserves （total proteins, total sugar, and total starch）, concen-trations of which were higher in fresh seed and declined as storage dura-tion increased. Germination was higher in fresh seeds and after 65 days, no germination was received perhaps due to deterioration of biochemi-cals in seeds. Patterns of seed deterioration were similar across all seed lots.

Acaricidal activity of Cymbopogon citratus and Azadirachta indica against house dust mites

Objective:To examine the acaricidal effects of the essential oil of Cymbopogon citratits leaf extract(lemongrass)and ethanolic Azadirachta indica leaf extract(neem)against house dust mites Dermatophagoides farinae(D.farinae)and Dermalophagoides pteronyssinus(D.pteronyssinus).Methods:Twenty-five adults mites were placed onto treated filter paper that is soaked with plant extract and been tested at different concentrations(50.00%,25.00%,12.50%,6.25%and 3.13%)and exposure times(24hrs,48hrs,72hrs and 96 hrs).All treatments were replicated 7 times,and the experiment repeated once.The topical and contact activities of the two herbs were investigated.Results:Mortalities from lemongrass extract were higher than neem for both topical and contact activities.At 50%concentration,both 24 hrs topical and contact exposures to lemongrass resulted in more than 91%mortalities for both species of inites.At the same concentration and exposure time,neem resulted in topical mortalities of 40.3%and 15.7%against D.pteronyssinus and D.farinae respectively;contact mortalities were 8.0%and 8.9%against the 2 mites,respectively.There was no difference in topical mortalities of D.pteronyssinus from exposure to concentrations oflemongrass and neem up to 12.50%;lemongrass was more effective than neem at the higher concentrations.Conclusions:Generally,topical mortalities of D.farinae due to lemongrass are higher than that due to neem.Contact mortalities of lemongrass are always higher that neem against both species of mites.

Antifertility effects of Azadirachta indica methanol seed extract on canine spermatozoa in vitro

Objective:To investigate the effects of Azadirachta(A.)indica methanol seed extract on canine male fertility in vitro.Methods:The effect of the A.indica methanol seed extract(2.5,5.0,10.0 and 20.0 mg/mL,respectively)on canine spermatozoa was evaluated in vitro at 1.0,2.5,5.0 and 10.0 min post-treatment.The two controls were comprised of sperm sample in phosphate-buffered saline,and sperm sample in phosphate-buffered saline and 1%dimethyl sulfoxide,respectively.The experiment was repeated(n=3)in duplicates with semen collected from 3 adult Basenjis.The effect of A.indica methanol seed extract treatments on canine spermatozoal total and progressive motility(phase-contrast microscopy),sperm vitality(eosin-nigrosin vital staining method)and total abnormalities(phase contrast microscopy,eosin-nigrosin staining and Papanicolaou staining methods)were determined.Results:A.indica methanol seed extract treatment caused a concentration and time-dependent decrease in sperm percentage total motility,progressive motility and vitality.The absence of sperm revival post-treatment provided evidence of a permanent or irreversible incapacitation of sperm by A.indica methanol seed extract.Conclusions:The inhibition of sperm motility and significant spermicidal effect observed post-treatment suggest that A.indica methanol seed extract may produce a potent antifertility or contraceptive effect in dogs if applied as an intra-vaginal cream pre-coitus.

New triterpenoids from the kernels of Azadirachta indica

Three new limonoids(1-3)and a new intact triterpenoid(4),along with three known constituents(5-7),were isolated from the dried kernels(after extracting azadirachtin)of Azadirachta indica.The structures of the new compounds 1-benzoyl-3-deacetyl-1-detigloyl salannin(1),7-tigloyl-12-oxo vilasini(2),azadiralactone(3)and azadirahemiacetal(4)were elucidated by means of spectroscopic analysis.The cytotoxities of these isolated constituents were assayed.

In vitro antifilarial activity of Azadirachta indica aqueous extract through reactive oxygen species enhancement

Objective:To evaluate an aqueous preparation from the Azadirachta indica leaves(AEA) against Setaria cervi(S.cervi),a model filarial parasite.Methods:In vitro efficacy of AEA was evaluated against S.cervi through estimation of relative motility value,dye exclusion test and MTT assay.Visible morphological alterations were monitored using conventional microscopic techniques in microfilariae and haematoxylin-eosin stained sections of AEA-treated adults.Results:Enhancement of reactive oxygen species in S.cervi treated with AEA was established through alteration in the activity of glutathione S-transferase,superoxide dismutase,catalase,peroxidase and level of superoxide anion and reduced glutathione.Conclusions:In vitro filaricidal activity of AEA is possibly through disturbing redox homeostasis by down-regulating and altering the level of some key antioxidants and regulatory enzymes like reduced glutathione,glutathione S-transferase,superoxide dismutase,catalase and glutathione peroxidase of S.cervi.

Pharmacodynamic evaluation for antiplasmodial activity of Holarrhena antidysentrica(Kutaja) and Azadirachta indica(Neemb) in Plasmodium berghei infected mice model

Objective:To investigate in-vivo anti-plasmodial activity of aqueous extracts of plants selected based on the symptomology mentioned in Ayurveda.Methods:The aqueous extracts of Holarrhena antidysentrica(H.antidysentrica)(Kutaja) and Azadirachta indica(A.indica)(Neemb) for their antiplasmodial potential in Plasmodium berghei(P.berghei) infected mice was assessed using Peters four day suppressive test.Both the extracts were administered at 2 dose levels,full dose(1 000 mg/d) and minimized dose(200 mg/d).10~6 P.berghei infected RBCs were injected on day ’0’ and treated from day ’0’ till day ’3’ post-infection,Tail blood smears were collected, giemsa stained and analyzed.The mice were observed for survival and parasitemia was assessed till 50%of mice in control survived.Results:It was observed that the percentage of parasitemia increased gradually in all the groups,with maximum in control group(Day 3-35,Day 9-46.98) and minimum in Chloroquine arm(Day 3-14.06.Day 9-19.92).The percentage of parasitemia was compared using Mann-Whitney U test depicting that all test groups exhibited reduction in parasitemia as compared to control(P-value【0.002 for all groups).These groups showed similar percentage of survival as Chloroquine.Conclusions:The present investigation demonstrated the anti-plasmodial effects of H.antidysentrica and A.indica.which are two most commonly used medicinal plants in Ayurved for treatment of fever.

Phytochemical Analysis and Antibacterial Activity of <i>Azadirachta indica</i>Leaf Extracts against <i>Escherichia coli</i>

Background: Traditional medicine has employed Azadirachta indica to treat a variety of ailments. However, there is little information on the use of this plant in Zambia. Objective: To evaluate the phytochemicals and antibacterial activity of Azadirachta indica leaf extracts against Escherichia coli. Materials and Methods: This was a laboratory-based experimental study in which ethanol and water were used for extraction by maceration. Phytochemical analysis was then done on the leaf extract. Using the disc diffusion method, varying concentrations of A. indica aqueous and ethanolic extracts were used to test the antibacterial activity of A. indica against Escherichia coli. The sensitivity of the tested microorganisms to aqueous and ethanolic leaf extracts was shown by zones of inhibition after incubation. Results: The phytochemical screening of the A. indica leaves revealed the presence of phenolics and tannins in both the ethanol and aqueous extract. Saponins, flavonoids and alkaloids were only present in the aqueous extract. While steroids were only present in the ethanol extract. The antibacterial activity of A. indica leaves extract was determined by zones of inhibition which showed that both aqueous and ethanol extracts had activity against E. coli. The Minimum Inhibitory Concentration was determined at 10 mg/mL for the aqueous extract and 20 mg/mL for the ethanol extract. The zones of inhibition increased with concentration. Conclusion: The extracts of A. indica displayed antibacterial activity against E. coli in a dose-dependent manner. Comparatively, the aqueous extract produced better antibacterial properties against E. coli than the ethanolic extract.

Ethanolic extract of Azadirachta indica ameliorates ovarian defects through phosphoinositide-3 kinase inhibition in a rat model of polycystic ovary syndrome

Objective:To assess the therapeutic potential of ethanolic extract of Azadirachta(A.)indica in rats with polycystic ovary syndrome(PCOS).Methods:Thirty-five prepubertal female Sprague Dawley rats were randomly divided into five groups with 7 animals in each group.Group 1 received 0.5%carboxy methyl cellulose orally.Groups 2 to 5 received testosterone propionate(0.2 mg/kg,s.c.)dissolved in olive oil daily for 42 days to induce PCOS.In addition,group 3 was administered with A.indica extract(100 mg/kg,0.5%carboxy methyl cellulose orally)from the 7th to 12th week,group 4 received quercetin(100 mg/kg,0.5%carboxy methyl cellulose orally)and group 5 received wartmannin(100 mg/kg,0.5%carboxy methyl cellulose orally).At the end of treatment,blood was collected for biochemical evaluation.Total follicular count and uterus corpus luteum count followed by PI3K gene expression in the ovary and uterus were evaluated.Results:The ethanolic extracts of A.indica significantly reduced body weight,ovary weight and uterus weight of rats.Extracts of A.indica also significantly increased the levels of serum glucose,total cholesterol,triglyceride,low-density lipoprotein,very low-density lipoprotein,insulin,testosterone,and luteinizing hormone.Treatment also reduced lipid peroxidation and increased antioxidant parameters in the liver homogenates of PCOS-induced rats.Histological examination of the ovary and uterus confirmed PCOS occurrence and remission state in the PCOS-induced and treated groups,respectively.Moreover,A.indica and quercetin significantly downregulated PI3K gene expression.Histopathological results of the ovary and uterus also proved the protective role of A.indica.Conclusions:A.indica leaf extract has beneficial effects in the treatment of PCOS by downregulation of PI3K gene expression.