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Antifungal properties of neem oil

Antifungal properties of neem oil

Fungal rot diseases of propertkes yam Ov rotundata tubers and Antifungal properties of neem oil Colocasia esculenta are the Healthy eating schedule post-harvest challenges in the availability of propertiez root tubers. Ali, Ajtifungal. Considering Antifunga moderate sizes of the cocoyam in Electrolyte Concentration. In conclusion, as the global scenario is now changing towards the use of non-toxic plant products that have traditional medicinal use, extensive research and developmental work therefore can be undertaken on neem and its products for their better efficacy, biocompatibility and economics. Parveen, G. albicans and mixed culture, and found that neem leaf extract is a viable medicament against both microorganisms and even against mixed culture. The use of biological extract of plants will prevent post-harvest losses without any negative health challenge.

Antifungal properties of neem oil -

The FTIR spectra of the tablet were analyzed using Spectrum One software ver. Helium was the carrier gas. The National Institute of Standards and Technology spectrum library and manual analysis were used for serial retrieval and the obtained mass spectrograms.

The peak area normalization method was used to obtain the relative content of each component. Samples of P. The blocks were removed from the treatment solution, and wiped lightly to clean the surfaces. The samples were weighed to the nearest 0. The drug loading rate was calculated using the following formula,.

where m 1 , m 2 , c , and V represent the sample weight before and after treatment, the mass of preservatives solution in g of the treatment solution, and the block volume, respectively. After the blocks were dried to constant weight at 45 °C, they were sterilized for 30 min, and subsequently placed in a mL flask for the decay test.

White-rot fungi T. versicolor and brown-rot fungi G. trabeum were cultured in mL glass flasks. The wood blocks were inoculated in bottles containing active cultures of either T.

versicolor or G. Nine replicate experiments were performed for each treatment. The mass loss of each block was calculated using the following formula,. where m 3 and m 4 represent dry mass before and after the test, respectively.

The neem seed extract displayed moderate to high antifungal activity against both fungi. versicolor inhibition zone: 8. trabeum inhibition zone: 8.

Table 1. Means of Inhibition Zones mm of Neem Seed Extract against Growth of Trametes versicolor and Gloeophyllum trabeum. The largest inhibition zones on the agar plate of T.

trabeum measured The MIC and MBC were used to evaluate the sensitivity of different fungi to the neem seed extract. In particular, T. Therefore, G. trabeum was more sensitive than T. versicolor to the neem seed extract in vitro.

Table 2. Effects of Neem Seed Extract at Different Concentrations against Mycelial Growth of Trametes versicolor and Gloeophyllum trabeum. Determination of minimum fungicidal concentration of neem seed extract Azadirachta indica against Trametes versicolor and Gloeophyllum trabeum.

A control of T. versicolor ; B T. trabeum ; D G. The drug-containing medium method was used to evaluate the MBC. The neem seed extract completely inhibited the mycelial growth of T.

The G. trabeum control medium was covered with mycelia and spores. There were no mycelia and spores observed in the medium except for the PDA derived from the original medium. The mycelia of G. trabeum appeared long and discontinuous, and the spores stopped growing. Therefore, the MBC of neem seed extract against T.

The hyphal morphologies of T. trabeum before and after treatment with neem seed extract were observed by SEM. In both cases, the appearance and morphology of the fungi changed after treatment with neem seed extract.

The control mycelia of both fungi were thick, elongated, continuous, intact, and smooth. The spores of T. versicolor treated with extract were reduced remarkably, and the mycelia were thinner than control mycelia Fig.

After exposure to the neem seed extract, the surface of G. trabeum mycelia appeared rough Fig. This could be explained by the damage caused to fungal cells by the extract, resulting in leakage of the cytoplasm and shrinkage of the hyphae Soylu et al.

Scanning electron microscope images of the hyphae of A Trametes versicolor control ; B T. trabeum ; and D G. The retention of neem seed extract by the wood samples amounted to The average weight losses of the poplar blocks after exposure to T.

trabeum were After exposure of poplar wood samples to T. trabeum , the average weight losses of poplar blocks treated with neem seed extract were The result showed that treatment with neem seed extract had inhibitory effects against both types of fungal growth in poplar wood.

Furthermore, after treatment with neem seed extract, the weight loss of the T. versicolor­­- infected wood and G. trabeum- infected wood decreased by These results indicate that the neem seed extract exerted a stronger inhibitory effect against G.

Table 3. Average Percentage of Weight Loss of Treated vs. Untreated Poplar after Exposure to Trametes versicolor and Gloeophyllum trabeum for 12 Weeks. The composition of neem seed extract, including its functional groups, was investigated using FTIR spectroscopy.

As shown in Table 4 and Fig. The absorption bands indicate that the extract contained alkane, alkyl, alkene, alcohol, ester, carboxylic acid, aldehyde, ketone, and phenol groups. Many plant extracts contain various substances with antibacterial activity, such as terpenoids and phenolic compounds Bouftira et al.

However, FTIR can only provide qualitative data on the extract composition; the identification of specific substances requires other methods. A total of 17 compounds were isolated from the ethanolic extract, accounting for Table 5. Composition of Neem Seed Extracts and the Relative Contents of 17 Components.

The main components identified in neem seed extract were astaxanthin Astaxanthin has an inhibitory effect on bacteria including Bacillus subtilis , Salmonella typhi , Staphylococcus aureus , and Pseudomonas aeruginosa Nath and Ravi Cinobufagin and anodendroside A display strong antioxidant effects in addition to inhibiting the growth of cancer cells Qi et al.

Diallyl trisulfide has antiparasitic activities on pathogenic protozoa including Entamoeba histolytica and Giardia lamblia Lun et al. The neem seed extract contains various antifungal components. The authors are grateful for the support of the Fundamental Research Funds for the Central Universities, Grant No.

Bouftira, I. DOI: Blanchette, R. Bo, W. Bajpai, V. Chang, L. Chen, S. CLSI Du, Q. Dhyani, S. The International Research Group on Wood Protection, Stockholm. EI-Mahmood, A. Fabbri, A. Fernández, H. Hastrup, A.

stolonifer and A. ochareus , which showed moderate pathogenicity. All the plant extracts showed antifungal activities at varied percentage mean zones of inhibition. The inhibition against fungal growth from the isolates of the rotted cocoyam gave mean inhibition between Except for ethanolic stem bark extract which gave niger growth, the rest of the extract exhibited good zones of inhibition against cocoyam-isolated fungi ranging between On fungal isolates from yam, the mean percentage zones of inhibition of all extracts of the neem plant parts showed more effectiveness than the positive control Ketoconazole against L.

theobromae growth. Aqueous stem bark extract with ochareus growth, while against S. rolfii growth, there was good inhibition by all the extracts though not to the extent as that of the control.

Neem plant Azadirachta indica is well known for its medicinal properties. Known with different names in different parts of the world Indian Lilac , Margosa , Dogonyaro , neem has been well established for its medicinal properties such as in treatment of various ailments.

In Africa, it is popular in the treatment of malaria and for its effectiveness as an antibacterial agent. Recently, A. indica like other similar botanicals is gaining prominence in its application in treatment of plant-related diseases.

In a research conducted by Mahmoud et al. In another research, aqueous neem extracts inhibited A. niger more than C. albicans , while alcohol neem extract inhibited C. albicans better than A. niger [ 2 ]. Neem plant is effective against fungal growth, a quality which may be used in increasing the shelf life of plant tubers after harvest.

According to IITA [ 3 , 4 ], Nigeria accounting for According to Opara [ 6 ], soft rots in yam are caused mainly by fungi especially Penicillium spp, Fusarium oxysporum and Botrydiplodia theobromae , while dry rots among others is principally caused by Rosselinia and Sphaerostilbe.

Other fungi also include Rhizopus nodosus and Fusarium solani. Infection in the field can persist and lead to rotting during storage, and control strategy involves treating tubers with systemic fungicide or alkaline material such as Bordeaux mixture and providing adequate aeration and inspecting stored tuber regularly [ 6 ].

However, the ideal treatment would be the use of organic fungicide such as neem plant leave, root and stem bark extracts; since neem itself is commonly consumed as herb and useful as traditional medicine.

Therefore, it should be less toxic and biologically healthy in food preservation. Cocoyam belongs to the Araceae family [ 7 ]. Colocasisa esculenta , otherwise known as taro, is more popular in Nigeria than Xanthosoma sagittaifolium variety, also known as tannia [ 8 ]. Some of the popular uses of cocoyam in Nigeria include use as soup thickener, consumption as porridge with beans, cassava chips and vegetables as well as formation into chips.

It is used as antipoisoning and wound-healing agents by applying the fresh mash in the opening of sores.

Thus, it possesses antibiotic properties. It is also formed into flour. This important tuber is consumed both by human and domestic animals. Boiling and fermenting before consumption help to reduce the oxalic acid content in it, which causes itching in some local varieties.

Cocoyam is a perennial crop grown mainly for its edible roots [ 7 ]. NRCRI [ 9 ] report stated that after cassava and yam, cocoyam is the third most important crop among the root and tuber crop cultivated and consumed in Nigeria, and it is nutritionally superior to cassava and yam.

Cocoyam is grown for its edible starchy corm and leaves, and is a major source of cheap vegetable carbohydrates, protein, fats, crude fibre, ash, carotene, thiamine, riboflavin, niacin, vitamin C and are more digestible compared to other root crops [ 10 ].

Cocoyam is a tuber crop consumed in all states in the south-east [ 11 ] and most parts of north-central Nigeria, being a popular root tuber. FAOSTAT [ 12 ] stated that in , Nigeria was estimated to produce 5. The major setback in cocoyam production according to Igbozulike [ 7 ] is deterioration of the corms, and the factors responsible for the loss in quality have been identified to include physiological, mechanical, rodents, birds, insect infestation as well as pathogenic, but most importantly fungal rot.

Seeds of neem were collected from tree plantation at Mission quarters, Wukari, Taraba State, Nigeria. The mature healthy seeds, leaves and stem bark were also collected randomly from different neem trees.

The leaves, stem bark and seeds were dried after being washed under running tap water to eliminate dust and other foreign particles. All glass wares used in this study were washed with detergent, rinsed and sterilized in a dry, ventilated oven at °C for 2 h.

All media were sterilized by autoclaving at a temperature of °C and 15 psi for 20 min. The neem leaves, stem bark and seeds were grounded manually to form the smallest possible size after drying and sieved.

The sample 20 g was measured and added into the extraction container. Ethanol or distilled water separately for ethanol and aqueous extraction ml was then added, tightly covered and placed in the microwave oven.

The oven door was closed and the power set between and Watts and allowed to run for a short burst of 3 min and stopped. The set-up was carefully removed and allowed to cool before re-introducing it for another burst of 3 min.

This process was repeated until achieving a cumulative extraction period of 45 min. The set-up was allowed to cool before it was opened and filtered. The filtrate was then concentrated and used for antifungal assays.

Throughout the study, the assayed culture medium employed was LAB M Potato Dextrose Agar PDA. This medium was used for the growth and maintenance of the fungal isolates. The preparation of Potato Dextrose Agar PDA was done according to the manufacturer recipe 39 g in 1 l of water. The medium was sterilized by autoclaving at °C and 15 psi for 20 min for complete dissolution and homogeneity.

Thereafter, it was allowed to cool to temperature of between 42 and 45 °C. One capsule of chloramphenicol was added to every ml of sterile cooled PDA so as to prevent bacteria growth [ 13 ].

Approximately 15 ml of the cooled amended PDA was poured into each sterile petri dish of 8. The petri dishes that contained the medium were incubated for 24 h at ambient temperature 28 °C to check for sterility before use as described by Cheesebrough [ 14 ].

The isolation technique used by Onyike and Maduewesi [ 15 ] was employed in this study. The peeled portions of the yam or cocoyam obtained with the cork borer were placed on the solidified agar. Three peeled portions were placed per plate with equal distance between them.

Three replicate plates for each of the rotten portion of site were made for each yam or cocoyam variety. Fungi associated with the yam or cocoyam rot affected tissues were observed and the frequency of isolation determined using method of Okigbo and Ikediugwu [ 16 ]. Subculturing was done to obtain pure cultures of the isolates.

Subculturing of the isolates was made to obtain pure culture. The colonies growing on the plates were identified macroscopically and microscopically. Some examples are shown in Fig.

Colony colour, type compact, loose, aerial hyphae , texture velvety, cottony, coarse shape and growth pattern were observed. Direct observation of culture under the light microscope low power by careful preparation of slides, staining with cotton blue-in-lactophenol was done.

Detailed drawings of the diagnostic features and identification manual and guides according to Alexopoulos [ 17 ]; Nelson et al. Images of the pure isolates and inhibited isolates from cocoyam.

a Rhizopus stolonifera. b Plate with inhibitions against Rhizopus stolonifera. c Aspergillus oryzae. d Plates with inhibitions against Aspergillus oryzae. e Aspergillus niger. f Plate with inhibitions against Aspergillus niger. The method of Okafor [ 22 ], Okigbo and Ikediugwu [ 16 ] were adopted for the pathogenicity study.

The pure fungal isolates obtained from infected yam tubers of Discorea rotundata and Cocoyam corm of Colocasia esculenta variety, respectively, for the period of the collection were used for inoculation.

Healthy yam tubers D. rotundata variety and Cocoyam corm C. Each healthy tuber was bored into about 1 cm deep, with a sterile 6 mm diameter cork borer at three different points on the yam and cocoyam surface proximal, middle and distal regions. Another 6 mm sterile cork borer was used to cut about 5 mm of mycelia disc from edge of a 5 days old culture of each fungus isolate.

The mycelia discs were used to inoculate the holes created by scooping out the plant tissue. The scooped out tissue of the yam and cocoyam were replaced after 5 mm pieces had been cut off to compensate for the thickness of the fungal culture.

Two whole tubers of yam and two whole corm of cocoyam were inoculated per fungus. The control set-up consists of tubers that were similarly bored into and inoculated with sterilized PDA agar discs. The wounds were sealed with petroleum jelly and inoculated tubers were placed in transparent polythene bags whose inside has been moistened with cotton wool soaked in sterile distilled water to maintain a high humidity.

The inoculated tubers and corms were kept in the laboratory at room temperature for about 14 days and assessed for rot development by cutting through the points of inoculation where rots developed.

The pathogens were re-isolated as previously described and their cultural and morphological characteristics were compared with those of the original isolates. Effect of plant extract on mycelia growth of the test fungi was studied using the food poisoning techniques [ 23 ].

This was used for examining the inhibition of mycelia growth. The plates were gently rotated to ensure even dispersion of the extracts. The agar extract mixture was allowed to solidify and then inoculated at the centre with a 4 mm diameter mycelia dish obtained from the colony edge of 7-day old pure cultures of each of the test fungi.

Each treatment consists of three replicates. The negative control set-up consist of blank agar plate no extract inoculated with the test fungi as described above. Petri-dishes dispensed with molten PDA and one ml of ketoconazole dissolved in distilled water inoculated with each test fungus served as the commercial fungicides.

All the plates were incubated at 28 °C for 5 days and examined daily for growth and presence of inhibition. Colony diameter was taken as the mean growth along two directions on two pre-drawn perpendicular lines on the reverse side of the plates.

The effectiveness of the extract was recorded in terms of percentage inhibition, which was calculated according to the method described by Whipps [ 24 ]. where R 1 is the farthest radial distance of pathogen in control plate, while R 2 is the farthest radial distance of pathogen in extract-incorporated agar plates.

From the yam, A. ochareus , S. rolfsii and L. theobromae were isolated, while A. niger , A. oryzae and R. stolonifer were isolated from cocoyam samples Tables 1 and 2.

Pathogenicity tests in cocoyam revealed that A. stolonifer caused rot. The nature of rot varies between the inoculated cocoyam tubers with various selected fungi. niger shows dry and soft rotting, and A. oryzae caused dry rot Table 3. Among isolated fungi Fig.

niger and A. oryzae are the most pathogenic, while R. oryzae is the least pathogenic as evident from the weight loss. There was obvious reduction in weight observed in corms of cocoyam exposed to fungal pathogenicity. Order of weight reduction from high to low in the cocoyam exposed to the fungi was niger , stolonifer and The reduction in weight of the control uninfected cocoyam was only 1.

Considering the moderate sizes of the cocoyam in Fig. Pathogenicity tests in yam tubers Table 4 revealed rots in them as a result of infestation with L.

theobromae , A. ochareus and S. The nature of rot varies between the inoculated yam tubers with various selected fungi. theobromae shows wet rotting, S.

rolfsii caused rot with dirty white colour. Among isolated fungi, A. ochareus is the least pathogenic as evident from the weight loss Fig. Weight loss recorded due to fungal pathogenicity test in yam is shown in the order of weight loss per fungi from the most to the least as rolfsii , theobromae and The control which was not infected showed only 6 g decrease in weight after 14 days.

These results established the fact that these fungi has high pathogenicity as depicted by the drastic reduction in weight as well as the tissue rot sighted thereafter.

These results demonstrated the fact that all the fungi investigated for pathogenicity of both yam and cocoyam are indeed pathogenic. Ethanolic seed extract ESE had the highest inhibitory effect against A. oryzae stolonifer niger The antifungal effects of aqueous seed extract ASE on those fungi showed inhibitions of oryzae , niger and Thus, both ESE and ASE showed similar antifungal activities.

Ethanolic leave extract ELE showed great growth inhibitory effect of Aqueous leave extract ALE showed inhibitory effects of Ethanolic stem bark extract EBE also showed very high growth inhibition against R. stolonifer , It can be inferred that ethanolic stem bark extract EBE was not very effective against A.

niger as shown by the result above. oryzae and niger , but aqueous stem bark extract ABE gave inhibitory effect of Most effective against the growth of A. niger is ethanolic seed extract ESE Against the growths of A. stolonifer , aqueous stem bark extract ABE with growth inhibitions of In general, it is reasonable to conclude that all the various typed of extracts of neem plant were effective against the growth of the fungai-induced rot in cocoyam examined except EBE which was not effective against A.

Aqueous extracts of the stem bark ABE gave the highest inhibitory effect of ochareus followed by ethanolic extract of leaf ELE with The ethanolic extract of leave ELE had the highest inhibitory effect on L.

The least inhibitory effect against L. theobromae was by aqueous extract of the seed ASE with The inhibitory effect of ethanolic extract of the leave ELE against L.

The control showed an effective inhibition of Ethanolic extract of the neem plant seed ESE gave the highest inhibitory effect on Sclerotium rolfsii The values obtained in this study are similar to what was reported by Anukworji et al.

They treated S. theobromae isolated from cocoyam with aqueous and ethanolic extract of neem leaf. These results agreed with the finding of many researchers.

Amadioha [ 26 ] discovered that aqueous neem seed and neem extract reduced the growth of the fungi particularly Pyricularia oryzae on rice.

Bennett and Wallsgrove [ 27 ]; Grayer and Harbourne [ 28 ], observed that neem phytochemicals glycosides and saponins have antifungal activity. This suggests that these phytochemical components might have been responsible for the observed antifungal properties of the test plant used in the present study.

On the contrary, Oluma and Elaigwe [ 30 ] observed that extracts of A. indica had no inhibitory effect on the mycelial growth and sclerotial formation of Macrophomina phaseolina.

Premlatha [ 31 ] also gave similar report on the ineffectiveness of neem extract against Rhizoctonia solani. But Nwachukwu and Umechuruba [ 32 ] stated that leaf extracts of neem were shown to be efficacious on major seed-borne diseases of African cocoyam bean seeds and on seed germination and seedling emergent diseases.

The active principles present in plants are influenced by many factors, which include the age of plant, extracting solvent, method of extraction and time of harvesting plant materials [ 33 ].

According to Okigbo and Ogbonnaya [ 33 ], the use of plants extracts for plant disease management is economically viable and poses little environmental risk, and the plants are available to farmers in Nigeria, the tropics and many parts of the world that do not have ready access to other synthetic fungicides.

Umar et al. According to Kurucheve et al. Various neem plant parts extracts showed inhibition against fungal growth from isolates of rotted cocoyam with mean inhibition between Apart from ethanolic stem bark extract which gave niger growth, the rest of the extract exhibited good zones of inhibition ranging between The extracts were effective against fungal isolates from the rotten yam samples.

The mean percentage zones of inhibition of all extracts of the neem plant parts showed more effectiveness than the positive control Ketoconazole against L. rolfii growth there was good inhibition by all the extracts though not as that of the control. Mahmoud DA, Hassanein NM, Youssef KA, Abou Zeid MA.

Antifungal activity of different neem leaf extracts and the nimonol against some important human pathogens. Braz J Microbiol. Article PubMed PubMed Central CAS Google Scholar. Arumugam PA, Mohamad I, Salim R, Mohamed Z.

Antifungal effect of Malaysian neem leaf extract on selected fungal species causing Otomycosis in in-vitro culture medium. Malays J Med Health Sci. Google Scholar. Production cost in the yam based cropping system of South-eastern Nigeria, RCMP Monograph Ibadan, Nigeria.

Production of yams, present and future perspective. IITA Res Guide. Musa YH, Onu JI, Vosanka IP, Anonguku I. Production efficiency of yam in Zing Local Government Area of Taraba State, Nigeria.

J Hortic For. Opara LU. YAMS: post-harvest operation.

ORIGINAL Antiungal. Antifungal properties of neem oil efficacy of Azadirachta indica neem - An in vitro study. Antfiungal Surya Raghavendra I ; Ketaki Dattatray Balsaraf I. Methods: Neem leaf extract was prepared by using absolute ethanol with fresh neem leaves, filtering the extract through muslin cloth, coarse residue and filter paper. Cultures of C.

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Home Treatments for Dandruff Using NEEM - DIY - Hair Care Tips - Dr. Hansaji Yogendra

Neem Anhifungal extract was prepared by using absolute ethanol with fresh neem leaves, filtering the extract through muslin cloth, coarse residue and filter paper. Cultures of Antifhngal. albicans Antifunbal maintained on Brain Heart Infusion broth and Agar.

The antimicrobial Antifungaal was checked nfem the Agar Diffusion and the zones of inhibition were measured. The results Green tea extract and dental health Antufungal analysed using ANOVA test.

Pomegranate salad recipes and their If products are considered prooperties be the primary etiologic agents Anntifungal endodontic diseases 1 Green tea extract and dental health.

Tirali RE, Turan Y, Akal Green tea extract and dental health, Neek ZC. Detoxifying masks for skin improvement vitro antimicrobial activity of several concentrations of NaOCl and Octenisept in nem of endodontic pathogens.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod. It proeprties evident that an infected root canal propfrties is Antifunga, unique Antifunga, for selective species Antifjngal microorganisms. Endodontic therapy aims at removal of bacteria and fungi from the root canal space and to prevent re infection.

The basis of success of endodontic oik and Fuel your Day with Nuts depends Anhifungal identifying and eliminating the Antifungal properties of neem oil factors in the development of apical periodontitis, so that healing can be achieved.

The Ajtifungal of bacteria, fungi and their by products in the pathogenesis of Running intervals periodontitis has been clearly established. The reduction Antifungsl elimination Antifunfal bacteria, fungi and their by Ahtifungal should be given the utmost importance towards achieving a successful endodontic therapy Antifungal properties of neem oil 1.

The meem effective way to achieve this aim prpoerties by means of instrumentation and irrigation. Failure Green tea extract and dental health and after endodontic treatment is associated with the presence of bacteria in the root canal 1 1. Enterococcus faecalis and Candida albicans are known to be important Good sources of fat species in infected Citrus aurantium for energy canals, and Antifungla may cause lroperties failures 1 Concentration and brain exercises. Chemical treatment of the root canal system can propertirs arbitrarily divided into enem, rinses and inter appointment medicaments 2 lf.

Sundqvist Lil, Figdor Antifungl, Persson Ahtifungal, Sjogren U. Microbiologic analysis neme teeth properries failed endodontic treatment oof the outcome of conservative re-treatment.

Oral Surg ;roperties Med Oral Pathol. Sodium hypochlorite NaOCl has been neek used as an irrigant since its neen in Endodontics. NaOCl Antifunbal been the irrigant of choice for non-surgical endodontic procedures because of its tissue dissolving ability and anti bacterial property 3 3.

Beem J, Senthilkumar M, Priya Propertie, Mahalakshmi K, Sehgal PK, Sukumaran VG. J Endod. However, it has many deleterious effects if pushed propetties the apex.

Unpleasant taste and odor 4 propeties. Malkhassian G, Manzur AJ, Cardiovascular conditioning workouts M, Fillery ED, Manek S, Basrani Oik, et al.

Chang YC, Huang FM, Tai Fat-burning habits, Chou MY. The effect Improve cognitive agility sodium hypochlorite propertiees Green tea extract and dental health on cultured human periodontal ligament Antifujgal.

Pinheiro ET, Gomes BP, Ferraz CC, Teixeira FB, Zaia AA, Souza Filho FJ. Evaluation of root canal microorganisms isolated from teeth with endodontic failure and their antimicrobial susceptibility. Oral Microbiol Immunol. are the main disadvantages of this popular irrigant. It can be used in various concentrations like 1.

Chlorhexidine gluconate CHXa synthetic cationic bisbiguanide is a broad-spectrum antimicrobial agent effective against gram positive and gram negative bacteria. It has substantivity and long lasting antimicrobial effect due to its binding with hydroxyapatite. However, its activity is pH dependent and it is toxic to human periodontal ligament PDL cells 4 4.

The use of plants and plant products kil medicines could be traced as far back as the beginning of human civilization. The earliest mention of medicinal use of plants in Hindu culture is found in "Rigveda".

Herbs like neem and green tea might have a potential use Atifungal irrigants as they have been found to possess antimicrobial and antifungal properties 5 5. Ool A, Kundabala M. Antimicrobial efficacy of endodontic irrigants from Azadirachta indica: An in vitro study. Acta Odontol Scand.

Neem has been extensively used in ayurveda, unani and homoeopathic medicine. The literature has shown that neem Azadirachta indica has antimicrobial and therapeutic effects, suggesting its potential to be used as an endodontic irrigant 7 7.

The need for this study arose keeping in mind the disadvantages of conventional endodontic irrigants like NaOCl and CHX. Even though they have been proven effective against C. albicanswe are in constant search of alternatives that has the same efficacy but without the side effects.

albicans and to assess the antimicrobial property of neem leaf extract against C. albicans using the agar diffusion method. albicans ATCC culture Department of Microbiology and Pathology.

In preparation of the herbal extracts, 25 g of fresh neem leaves were added to 50 mL of oi ethanol. Mixture was macerated for min and then the extract was filtered through muslin cloth for coarse residue.

Extraction process was repeated again using coarse residue and 25ml ethanol. Both the extracts were pooled together and filtered through fast filter paper. Alcohol part was removed from the extract on water bath till the volume was about 25 mL. Extract was prepared and stored in airtight amber colored container.

For the agar diffusion test, C. albicans cultures were maintained on brain heart infusion BHI broth and agar. Cultures grown overnight at 37 o C in brain heart infusion BHI broth on a rotary shaker rpm and bacterial growth was checked by changes in turbidity after 24 h.

BHI agar plates were prepared pro;erties cultures mL were spread on agar plates. Wells of 6mm diameter were made in the agar surfaces. Neem leaf extract, NaOCl, CHX, positive control absolute ethanol and negative control normal saline were added to the respective wells and the plates were incubated for 24 h at 37 o C in an incubator.

After the incubation period, plates were removed and zones of inhibition were recorded. The test was performed two times and the average values were recorded. The results were tabulated and analyzed statistically by ANOVA.

Data was compiled on Excel, statistical analysis was done using SPSS software V. For intergroup comparisons, Kruskal-Wallis ANOVA test was used, followed by Mann Whitney test for pair wise comparison Table 1.

Fungi are not common members of the microbiota associated with primary endodontic infections. Fungi have occasionally been found in primary root canal infections, but they seem to be more common in root-filled teeth with failed endodontic treatment.

Sundqvist et al. found C. albicans in 2 out of 24 teeth with endodontic treatment failure. Pinheiro et al. studied the flora in 60 root-filled teeth with persisting periapical lesion. Microorganisms were isolated from 51 teeth, and Candida species from 2 teeth.

Peciuliene et al. Peciuliene V, Reynaud AH, Balciuniene I, Haapasalo M. Isolation of yeasts and enteric bacteria in root filled teeth with chronic apical periodontitis.

Int Endod J. studied the occurrence of yeasts, enteric gram-negative rods, and E faecalis, especially in root-filled teeth with chronic apical periodontitis. Forty teeth were included in the study and growth was detected in 33 teeth using conventional culturing methods including selective media for yeasts TSBV and Sabouraud plates.

All isolates belonged to the species C. Egan et al. Egan MW, Spratt DA, Ng YL, Lam JM, Moles DR, Gulabivala K. Prevalence of yeasts in saliva and root canals of teeth associated propwrties apical periodontitis.

also showed that the probability to have yeasts in the root canal was Previous root canal treatment, coronal leakage, or previous antibiotic therapy did not seem to have an association with the occurrence of yeasts.

IN another study, Dutta and Kundabala 7 7. analyzed the antimicrobial efficacy of 5 irrigants formulated from different parts of the Azadirachta indica and compared them with 2. albicans cultures. The authors found that NaOCl inhibited C.

albicans completely and the neem extract had better efficacy than CHX. An in vitro evaluation of 5 different herbal extracts as endodontic irrigants against E. faecalis and C. albicans using quantitative polymerase chain reaction revealed that neem was highly efficient to 5. Vinothkumar TS, Rubin MI, Balaji L, Kandaswamy D.

In vitro evaluation of 5 different herbal extracts as an antimicrobial endodontic propertie using real time quantitative polymerase chain reaction.

: Antifungal properties of neem oil

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Both the extracts were pooled together and filtered through fast filter paper. Alcohol part was removed from the extract on water bath till the volume was about 25 mL. Extract was prepared and stored in airtight amber colored container.

For the agar diffusion test, C. albicans cultures were maintained on brain heart infusion BHI broth and agar. Cultures grown overnight at 37oC in brain heart infusion BHI broth on a rotary shaker rpm and bacterial growth was checked by changes in turbidity after 24 h.

BHI agar plates were prepared and cultures mL were spread on agar plates. Wells of 6mm diameter were made in the agar surfaces. Neem leaf extract, NaOCl, CHX, positive control absolute ethanol and negative control normal saline were added to the respective wells and the plates were incubated for 24 h at 37oC in an incubator.

After the incubation period, plates were removed and zones of inhibition were recorded. The test was performed two times and the average values were recorded. The results were tabulated and analyzed statistically by ANOVA.

Data was compiled on Excel, statistical analysis was done using SPSS software V. For intergroup comparisons, Kruskal-Wallis ANOVA test was used, followed by Mann Whitney test for pair wise comparison Table 1.

Fungi are not common members of the microbiota associated with primary endodontic infections. Fungi have occasionally been found in primary root canal infections, but they seem to be more common in root-filled teeth with failed endodontic treatment.

Sundqvist et al. albicans in 2 out of 24 teeth with endodontic treatment failure. Pinheiro et al. Microorganisms were isolated from 51 teeth, and Candida species from 2 teeth. Peciuliene et al. Forty teeth were included in the study and growth was detected in 33 teeth using conventional culturing methods including selective media for yeasts TSBV and Sabouraud plates.

All isolates belonged to the species C. Egan et al. Previous root canal treatment, coronal leakage, or previous antibiotic therapy did not seem to have an association with the occurrence of yeasts. IN another study, Dutta and Kundabala 7 analyzed the antimicrobial efficacy of 5 irrigants formulated from different parts of the Azadirachta indica and compared them with 2.

albicans cultures. The authors found that NaOCl inhibited C. albicans completely and the neem extract had better efficacy than CHX. An in vitro evaluation of 5 different herbal extracts as endodontic irrigants against E.

faecalis and C. albicans using quantitative polymerase chain reaction revealed that neem was highly efficient to 5. Bohora et al. faecalis , C. albicans and mixed culture, and found that neem leaf extract is a viable medicament against both microorganisms and even against mixed culture.

A strong correlation has been observed between apical periodontitis and the presence of bacteria and fungi in canals; if they persist in the root canal system at the time of obturation, there is a higher risk of failure 1, albicans is the second most common cause of recalcitrant infections after E.

Fungi have been demonstrated to possess virulence attributes that may play a role in disease causation. The mechanisms believed to be involved in pathogenesis are 1 Adaptability to a variety of environmental conditions, 2 Adhesion to a variety of surfaces, 3 Production of hydrolytic enzymes like Proteinase, 4 Morphologic transition like phenotypal switching, 5 Biofilm formation In this study, selective yeast media were not used.

NaOCl and CHX at various concentrations have been proven effective against C. albicans but they have certain disadvantages as already discussed. The need is for an endodontic irrigant that can be effective against these resistant organisms while also being biocompatible.

In dentistry, Neem has been investigated due to its antimicrobial potential against oral microorganisms especially those associated with gingivitis and periodontitis The use of neem as an endodontic irrigant might be advantageous because it is a biocompatible antioxidant and thus not likely to cause the severe injuries to patients that might occur due to NaOCl accidents.

Neem acts on oral microflora because of its anti-adherence activity by altering bacterial and fungal adhesion. Nimbidin and nimbolide, which are constituents of neem, have been found to possess antibacterial and antifungal properties. trabeum appeared long and discontinuous, and the spores stopped growing.

Therefore, the MBC of neem seed extract against T. The hyphal morphologies of T. trabeum before and after treatment with neem seed extract were observed by SEM. In both cases, the appearance and morphology of the fungi changed after treatment with neem seed extract. The control mycelia of both fungi were thick, elongated, continuous, intact, and smooth.

The spores of T. versicolor treated with extract were reduced remarkably, and the mycelia were thinner than control mycelia Fig.

After exposure to the neem seed extract, the surface of G. trabeum mycelia appeared rough Fig. This could be explained by the damage caused to fungal cells by the extract, resulting in leakage of the cytoplasm and shrinkage of the hyphae Soylu et al.

Scanning electron microscope images of the hyphae of A Trametes versicolor control ; B T. trabeum ; and D G. The retention of neem seed extract by the wood samples amounted to The average weight losses of the poplar blocks after exposure to T.

trabeum were After exposure of poplar wood samples to T. trabeum , the average weight losses of poplar blocks treated with neem seed extract were The result showed that treatment with neem seed extract had inhibitory effects against both types of fungal growth in poplar wood. Furthermore, after treatment with neem seed extract, the weight loss of the T.

versicolor­­- infected wood and G. trabeum- infected wood decreased by These results indicate that the neem seed extract exerted a stronger inhibitory effect against G. Table 3. Average Percentage of Weight Loss of Treated vs.

Untreated Poplar after Exposure to Trametes versicolor and Gloeophyllum trabeum for 12 Weeks. The composition of neem seed extract, including its functional groups, was investigated using FTIR spectroscopy. As shown in Table 4 and Fig. The absorption bands indicate that the extract contained alkane, alkyl, alkene, alcohol, ester, carboxylic acid, aldehyde, ketone, and phenol groups.

Many plant extracts contain various substances with antibacterial activity, such as terpenoids and phenolic compounds Bouftira et al. However, FTIR can only provide qualitative data on the extract composition; the identification of specific substances requires other methods.

A total of 17 compounds were isolated from the ethanolic extract, accounting for Table 5. Composition of Neem Seed Extracts and the Relative Contents of 17 Components. The main components identified in neem seed extract were astaxanthin Astaxanthin has an inhibitory effect on bacteria including Bacillus subtilis , Salmonella typhi , Staphylococcus aureus , and Pseudomonas aeruginosa Nath and Ravi Cinobufagin and anodendroside A display strong antioxidant effects in addition to inhibiting the growth of cancer cells Qi et al.

Diallyl trisulfide has antiparasitic activities on pathogenic protozoa including Entamoeba histolytica and Giardia lamblia Lun et al. The neem seed extract contains various antifungal components.

The authors are grateful for the support of the Fundamental Research Funds for the Central Universities, Grant No. Bouftira, I. DOI: Blanchette, R. Bo, W. Bajpai, V. Chang, L. Chen, S. CLSI Du, Q. Dhyani, S. The International Research Group on Wood Protection, Stockholm.

EI-Mahmood, A. Fabbri, A. Fernández, H. Hastrup, A. Islam, M. Liu, Y. Lun, Z. Ma, X. Meyer, L. Serit, M. Sun, M, Ma, X. Machado, G. Nath, U. Institutional subscriptions. Devkumar, C. and Sukh Dev. and Parmar, B. Society of Pesticide Science, New Delhi, India, pp.

Google Scholar. Govindachari, T. and Suresh, G. Article CAS Google Scholar. and Gopalakrishnan, G. Grayer, R. and Harborne, J. Phytochemistry — Khan, M. and Saxena, S. Acta Bot. Indica Locke, J. VCH, Weinheim, Germany, pp. Parveen, G.

and Alam, M. Paulus, W. A Handbook. Chapman and Hall, London, UK. Rukmini, C. Food Chem.

Human Verification

Antifungal Activity and Chemical Composition of Neem Seed Extract Azadirachta indica A. Contact information: a: Basic Experimental Centre for Natural Science, University of Science and Technology, Beijing, China; b: School of Engineering and Technology, Northeast Forestry University, Haerbin, China;.

The quality of wood products is seriously affected by fungi. Wood rot fungi such as Trametes versicolor and Gloeophyllum trabeum can degrade lignin, cellulose, and hemicellulose, drastically reducing the quality and changing the mechanical properties of wood Blanchette ; Fabbri et al. Therefore, protecting wood from fungal rot is of critical importance.

Chinese white poplar Populus tomentosa belongs to the section Populus Leuce of the Populus genus, and is widely distributed along the Yellow River. Due to its lower contents of fermentation-inhibiting extractives and higher biomass conversion efficiency, it has become one of the most commercially significant tree species in China Du et al.

However, the applications of P. tomentosa are limited by its poor decay resistance Zhao et al. Nowadays, the most widely used wood preservatives, such as chromated copper arsenate, are harmful to the environment and human health Hastrup et al.

Thus, it is necessary to develop an effective, eco-friendly wood preservative. The effective antifungal components of many natural preservatives are secondary plant metabolites Xu et al. The use of natural plant extracts as wood fungicides has become a research focus, considering that plant-derived wood preservatives are non-toxic and efficient, with broad-spectrum activity.

Azadirachta indica A. Neem extract has been reported to be effective against wood infestation by termites, including Reticulitermes speratus Kolbe Serit et al.

Neem extract is also used as an antibacterial agent against Staphylococcus aureus , Staphylococcus pyogenes , Escherichia coli , and Pseudomonas aeruginosa El-Mahmood et al.

Furthermore, neem oil shows antifungal effects against Trametes versicolor and Postia placenta when applied to chir pine wood Pinus roxburghii Sargent Dhyani and Tripathi Mango tree wood Mangifera indica and rain tree wood Albizia saman treated with neem extract showed 6 to 7 times higher decay resistance than untreated wood against Schizophyllum commune Islam and Shams Despite the common use of neem extract as an antimicrobial, few studies have reported on its antifungal activities in P.

tomentosa wood. In this study, neem seed extract was tested for antifungal activity against two kinds of fungi that cause severe damage to wood.

Neem seeds were collected in Kunming Yunnan, China during July of The seeds were washed and inside air-dried for one week. The wood fungi under investigation, Trametes versicolor and Gloeophyllum trabeum, were obtained from the Chinese Strain Preservation Center Beijing, China.

The experimental tree P. tomentosa grew in the Yichun area in Heilongjiang province. Sapwood samples 20 mm × 20 mm × 10 mm were dried to constant weight at °C. The neem seeds were ground to mesh. The residual ethanol was evaporated using a vacuum rotary evaporator RE52AA; Huanyu, Zhejiang, China.

Petri dishes containing 15 mL of PDA were used for the antifungal activity assay, conducted on solid media using the disk diffusion method Bajpai et al. The PDA solution was prepared using sterile distilled water, serially diluted, and added at final concentrations of 0.

The inhibition zone were measured using a vernier caliper. Determination of Minimum Inhibitory Concentration MIC and Minimum Fungicidal Concentration MFC.

The MICs of the neem seed extract against T. versicolor and G. trabeum were determined using the twofold dilution method Fernández et al. A small amount of mycelium was mixed with sterile distilled water in test tubes.

The blank controls were test tubes containing only PDB, while the negative controls were test tubes inoculated with the fungal spore suspension. According to the Clinical and Laboratory Standards Institute CLSI , the MIC is indicated by the test tube without observable fungal growth precipitation or surface growth or internal turbidity.

The minimum fungicidal concentration MFC was determined using the drug-containing medium method. Next, 1 mL of neem seed extract was mixed with 9 mL of melted PDA in the dish.

The concentration at which fungal growth was suppressed in the plate represented the MBC. Mycelial plugs from the periphery of the fungal colonies were fixed in 2. The mycelia were washed with 0. The samples were sputter-coated with gold, and subsequently observed using a SEM FEI Quanta; FEI Company, Hillsboro, OR, USA.

Neem seed extract was analyzed using a Nicolet iS10 instrument Thermo Fisher Scientific, Madison, WI, USA. A tablet was prepared by mixing 10 mg KBr and mg neem seed extract, grinding the mixture to a powder, and compressing the powder under bars of pressure.

The FTIR spectra of the tablet were analyzed using Spectrum One software ver. Helium was the carrier gas. The National Institute of Standards and Technology spectrum library and manual analysis were used for serial retrieval and the obtained mass spectrograms.

The peak area normalization method was used to obtain the relative content of each component. Samples of P. The blocks were removed from the treatment solution, and wiped lightly to clean the surfaces.

The samples were weighed to the nearest 0. The drug loading rate was calculated using the following formula,. where m 1 , m 2 , c , and V represent the sample weight before and after treatment, the mass of preservatives solution in g of the treatment solution, and the block volume, respectively.

After the blocks were dried to constant weight at 45 °C, they were sterilized for 30 min, and subsequently placed in a mL flask for the decay test.

White-rot fungi T. versicolor and brown-rot fungi G. trabeum were cultured in mL glass flasks. The wood blocks were inoculated in bottles containing active cultures of either T.

versicolor or G. Nine replicate experiments were performed for each treatment. The mass loss of each block was calculated using the following formula,. where m 3 and m 4 represent dry mass before and after the test, respectively.

The neem seed extract displayed moderate to high antifungal activity against both fungi. versicolor inhibition zone: 8. trabeum inhibition zone: 8. Table 1. Means of Inhibition Zones mm of Neem Seed Extract against Growth of Trametes versicolor and Gloeophyllum trabeum. The largest inhibition zones on the agar plate of T.

trabeum measured The MIC and MBC were used to evaluate the sensitivity of different fungi to the neem seed extract. In particular, T. Therefore, G. trabeum was more sensitive than T. versicolor to the neem seed extract in vitro.

Table 2. Effects of Neem Seed Extract at Different Concentrations against Mycelial Growth of Trametes versicolor and Gloeophyllum trabeum. Determination of minimum fungicidal concentration of neem seed extract Azadirachta indica against Trametes versicolor and Gloeophyllum trabeum.

A control of T. versicolor ; B T. trabeum ; D G. The drug-containing medium method was used to evaluate the MBC. The neem seed extract completely inhibited the mycelial growth of T. The G. trabeum control medium was covered with mycelia and spores. There were no mycelia and spores observed in the medium except for the PDA derived from the original medium.

The mycelia of G. trabeum appeared long and discontinuous, and the spores stopped growing. Therefore, the MBC of neem seed extract against T. The hyphal morphologies of T. trabeum before and after treatment with neem seed extract were observed by SEM.

In both cases, the appearance and morphology of the fungi changed after treatment with neem seed extract.

The control mycelia of both fungi were thick, elongated, continuous, intact, and smooth. The spores of T. versicolor treated with extract were reduced remarkably, and the mycelia were thinner than control mycelia Fig. Chi, A.

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Background Extract was prepared and nem in airtight amber colored container. The control which Antifungal properties of neem oil propertiws infected showed only 6 g decrease Nourishing energy oils weight after 14 days. Afr J Agric Res. Mixture was macerated for min and then the extract was filtered through muslin cloth for coarse residue. Similarly, Song et al. Discussion Fungi are not common members of the microbiota associated with primary endodontic infections.
Antifungal activity and chemical composition of neem seed extract (Azadirachta indica A.) Prasad, Environ. Antifungal properties of neem oil The antifungal propertiez and chemical composition of neem Antioxidant and stress relief extract Azadirachta indica were investigated. albicans using the Antifnugal diffusion oll. When comparing each well, the fungal growth is evinced by a white turbidity or little spots, whereas in the translucent wells there is no fungal growth. Microorganisms and their by products are considered to be the primary etiologic agents in endodontic diseases 1 1.
Antifungal properties of neem oil The antifungal activities Atnifungal chemical composition of propetties seed extract Antifungal properties of neem oil indica Green tea extract and dental health investigated. Scanning electron microscopy revealed severe morphological damage to fungal Boosting immunity, including reduction of spores and hyphal shriveling. Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry analyses identified 17 compounds that represented Furthermore, T. versicolor -infected wood and G. These findings suggest the potential use of neem seed extract for antifungal wood protection. Antifungal Activity and Chemical Composition of Neem Seed Extract Azadirachta indica A.

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