Avaliação hematológica e hepática de ovinos sob tratamento com torta de neem (Azadirachta indica) / Hematological and liver evaluation of sheep under treatment with neem pie (Azadirachta indica)

Devido aos altos índices de resistência dos parasitas aos diferentes princípios ativos comerciais, novas alternativas de controle vêm sendo estudadas, entre elas a fitoterapia. Essas medidas visam a busca de métodos auxiliares no controle das parasitoses, entretanto, muitos produtos estão disponíveis no mercado e não têm comprovação científica de sua eficácia ou de possíveis efeitos colaterais. O objetivo deste estudo foi avaliar os efeitos hematológicos e hepáticos após a administração de torta de Neem (Azadirachta indica) em ovinos. Foram testadas três dosagens da torta de Neem adicionada ao sal mineral (1, 2 e 4%), administradas por 126 dias para 32 ovinos da raça Lacaune, divididos em quatro grupos sendo três grupos para os diferentes tratamentos e um controle, o qual recebeu somente sal mineral. Amostras de sangue foram colhidas a cada 21 dias para realização do hemograma completo, dosagem de proteína plasmática total e fibrinogênio e avaliação da bioquímica clínica hepática. Nestas mesmas ocasiões, amostras de fezes foram coletadas para a quantificação de ovos por grama de fezes (OPG). Foram observadas diferenças estatísticas entre momentos e grupos para diversas variáveis, porém sem estarem relacionadas à administração de torta de Neem. Os resultados obtidos de hemograma completo, dosagem de proteína plasmática total, fibrinogênio e de bioquímica clínica hepática indica

Due to high levels of parasite resistance to different commercial active ingredients, new control alternatives are being studied, including the phytotherapy. These measures aim to search for auxiliary methods in the control of parasitic diseases. However, there are many products available in the market and there are no scientific proof of its efficacy. The objective of this study was to evaluate the hematological and hepatic effects following administration of Neem pie (Azadirachta indica) in sheep. Three concentrations of Neem cake was added to the mineral salt (1, 2 and 4 %) and administered during 126 days to 32 Lacaune breed sheeps, divided into four groups: three groups for different treatments and a and a control were tested, the ladder receiving only mineral salt. Blood samples were taken every 21 days to perform the complete blood count, serum total plasma protein and fibrinogen and liver biochemical evaluation. In those same times, fecal samples were collected for quantification of eggs per gram of feces (EPG). Statistical differences between times and groups for several variables were observed, but without being related to the administration of Neem pie. The results of complete blood count, measurement of total plasma protein, fibrinogen and hepatic clinical biochemistry indicated that administration of Neem pie at concentrations of 1, 2 and 4%, does not interfere in hematological values, or on the integrity and liver function Lacaune sheep breed.

Insecticidal activities and mechanism of extracts from neem leaves against Oxya chinensis / Atividade e mecanismos inseticidas de extratos de folhas "neem" contra Oxya chinesis

In this study, neem leaves were successively extracted with petroleum ether, 95% ethanol and water and the insecticidal activities of these extracts against Oxya chinensis larvae were measured. The results showed that 95% ethanol extract gave the highest extraction yield and insecticidal activity, and it was further extracted with five different solvents. The petroleum ether extract from the 95% ethanol extract possessed the highest insecticidal activity with median lethal concentration values ranging from 14.93 to 55.66mg/mL. The gas chromatography-mass spectrometer analysis showed that the petroleum ether extract mainly composed of alkanes, olefin, esters and amide. The pathological examination revealed that the prominent lesions, including reduced regenerative cells in midgut and swelled and degenerated cylindrical cells, were observed in the 5th instar Oxya chinensis after treatment. The ultrastructural features showed that the cylindrical cells, microvilli and mitochondria were seriously damaged. These results suggested that the petroleum ether extract from neem leaves had potent insecticidal activity and could be a candidate insecticide.(AU)

Nesse estudo, folhas "neem" foram extraídas com sucesso com éter de petróleo, 95% de etanol e água, e as atividades inseticidas desses extratos foram medidas contra larvas de Oxya chinesis. Os resultados mostram que extrato com 95% de etanol deram o maior resultado de extração e atividade inseticida e foi então extraído utilizando mais cinco diferentes solventes. O éter de petróleo do extrato de 95% etanol apresentou maior atividade inseticida com concentração letal média variando de 14.93 a 55.66mg/mL. A análise por cromatografia de massa mostrou que o extrato de éter de petróleo está composto principalmente de alcanos, alcenos, ésteres e amidas. A avaliação patológica revelou que as lesões proeminentes, inclusive células regenerativas reduzidas no intestino e células cilíndricas edemaciadas e degeneradas foram observadas no quinto estágio de desenvolvimento da Oxya chinesis após tratamento. As características ultraestruturais mostraram que as células cilíndricas, microvilos e mitocôndrias apresentavam lesões graves. Esses resultados sugerem que o extrato de éter de petróleo de folhas de "neem" tem atividade inseticida potente e pode ser um candidato a inseticida.(AU)

Insights on germinability and desiccation tolerance in developing neem seeds (Azadirachta indica): Role of AOS, antioxidative enzymes and dehydrin-like protein.

The germinability and desiccation tolerance (DT) in developing seed are regulated by cellular metabolism involving active oxygen species (AOS) and protective proteins during maturation drying. The aim of the present investigation was to unravel the functions of AOS (superoxide, H2O2 and OH-radical), antioxidative enzymes (SOD, CAT and APX) and dehydrin-like proteins in regulating the germinability and DT in undried and artificially desiccated developing neem seeds. Germination was first observed in seeds of 8 weeks after anthesis (waa) whereas DT was noticed from 9 waa. High levels of superoxide in undried and artificially desiccated seeds of 9 waa were rapidly declined up to 15 waa with simultaneous increase in levels of SOD (quantitative and isoenzymes) that dismutates superoxide with corresponding formation and accumulation of H2O2. Activities and isoenzymes of APX and CAT were promoted in seeds from 9 to 12 waa. Intensity of dehydrin-like proteins increased as development progressed in seeds with higher intensities in slow dried (SD) seeds. Desiccation modulated the metabolism for the acquisition of germinability and DT in the developing neem seeds from 8 to 15 waa by altering the levels of superoxide, H2O2 and OH-radical those possibly act as signalling molecules for reprogramming protective proteins. Desiccation mediated the expression of new bands of SOD and APX in undried as well as SD seeds during 9-12 waa but the bands were more intense in SD seeds. The superoxide and H2O2-regulated intensity of dehydrin-like protein in SD seeds further validated our conclusion.

Morphological and biochemical changes in Azadirachta indica from coal combustion fly ash dumping site from a thermal power plant in Delhi, India.

The foliar and biochemical traits of Azadirachta indica A. Juss from fly ash (FA) dumping site in Badarpur thermal power plant (BTPP) New Delhi, India was studied. Three different experimental sites were selected at different distances from the thermal power plant. Ambient suspended particulate matter (SPM) and plant responses such as leaf pigments (chlorophyll a, chlorophyll b, and carotenoids), total chlorophyll, net photosynthetic rate, stomatal index (SI), stomatal conductance (SC), intercellular carbon dioxide concentration [CO2]i, net photosynthetic rate (NPR), nitrogen, nitrate, nitrate reductase activity, proline, protein, reducing sugar and sulphur content were measured. Considerable reduction in pigments (chlorophyll a, chlorophyll b and carotenoids), and total chlorophyll was observed at fly ash dumping site. Fly ash stress revealed the inhibitory effect on Nitrate reductase activity (NRA), Nitrate, soluble protein, and reducing sugar content, whereas stimulatory effect was found for the stomatal index, nitrogen, proline, antioxidants and sulphur content in the leaves. Under fly ash stress, stomatal conductance was low, leading to declining in photosynthetic rate and increase in the internal CO2 concentration of leaf. Single leaf area (SLA), leaf length and leaf width also showed a declining trend from control to the polluted site. Antioxidant enzymes increased in leaves reflecting stress and extenuation of reactive oxygen species (ROS).

Antimicrobial effect of ethanol extract of leaf neem (Azadirachta indica A. Juss) on Listeria monocytogenes

Listeria monocytogenes es un patógeno causante de enfermedades alimentarias. En la búsqueda de controlar su propagación utilizando sustancias naturales se planteó el objetivo de mostrar si el extracto etanólico foliar de neem (Azadirachta Indica A. Juss.) tiene efecto antimicrobiano sobre L. monocytogenes ICTA-12446. El extracto se obtuvo a partir de hojas de neem sometidas a secado por 8 días, se redujeron de tamaño mecánicamente, se sometieron a maceración en frío por 3 días usando etanol 96% en recipientes ámbar, se filtró y concentró en rota evaporador. Se estandarizó el concentrado con dimetilsulfóxido (DMSO) a una concentración de 60 mg/L. Listeria monocytogenes ICTA-12446, fue inoculado en caldo nutriente junto con soluciones del extracto a diferentes concentraciones (20, 30, 40, 50 y 60 mg/L), se emplearon tiempos de contacto (2.5, 5, 10 y 15 minutos). Cumplido cada tiempo se realizaron diluciones seriadas e inocularon en agar nutritivo por extensión durante 24 h a 37ºC. Se efectuó el recuento en Unidades Formadoras de Colonias UFC. Al comparar las concentraciones del extracto se evidencia entre 20 y 60 mg/mL diferencia significativa, mientras que en 30, 40 y 50 mg/mL un comportamiento similar. Al contrastar tiempos de contacto, se observa que entre el tiempo 2.5 min y los restantes un p=0,03. El tiempo mínimo donde existió inhibición fue 2.5 minutos, y concentración mínima inhibitoria de 20 mg/mL. Los cuatro tiempos de contacto arrojan porcentajes de inhibición microbiana de 100% al emplear 60mg/mL. Se concluye que el extracto etanólico foliar de neem posee un efecto inhibitorio sobre Listeria monocytogenes(AU)

Listeria monocytogenes is a pathogen causing foodborne illness. In seeking to control its spread using natural substances in order to show if the leaf ethanol extract of neem (Azadirachta indica A. Juss) has antimicrobial effect on L. monocytogenes ICTA-12446, was proposed. The extract was obtained from neem leaves, which was subjected to drying for 8 days. It was reduced in size mechanically, and subjected to cold soak for 3 days, using 96% ethanol in amber vessels, filtered and concentrated in rot evaporator. Concentrated was solubilized with dimethylsulfoxide (DMSO) and standarized to achieve a concentration of 60 mg/mL Listeria monocytogenes was inoculated in nutrient broth with extract solutions at different concentrations (20, 30, 40, 50 and 60mg/mL), four contact times (2.5, 5, 10 and 15 minutes) were used. Completed each time it was diluted and inoculated on nutrient agar by extension for 24h at 37ºC. The count of Colony Forming Units UFC was taking. Comparing the concentrations of the extract between 20 and 60mg /mL significant difference was appreciate, while 30, 40 and 50 mg/mL show a similar behavior. Contrasting contact times observed between time 2.5 min and the remaining p = 0.03. The minimum time where there was some kind of inhibition was 2.5 minutes, and minima inhibitory concentration of 20mg/mL. The four contact times yield microbial inhibition percentages of 100% by using 60mg/L. It is concluded that ethanol extract of neem leaf has an inhibitory effect on L. monocytogenes(AU)

Role of chloroplastidial proteases in leaf senescence.

In this report the effect of hydrogen peroxide (H2O2) on peroxidase (POD) activity during leaf senescence was studied with and without phenylmethylsulfonyl fluoride (PMSF) pre-treatment in detached neem (Azadirachta indica A. juss) leaf chloroplasts. Increased POD activity was detected in natural and H2O2-promoted senescent leaf chloroplasts compared to untreated control mature green leaf chloroplasts. However, under H2O2 POD activity markedly increased at 1 day, and then significantly decreased until 4 days. In the presence of H2O2, PMSF, the induction of POD activity was alleviated at 1 day, whereas reduced after 4 days. In contrast, in the presence of H2O2, cycloheximide (CX), the induction of POD activity was reduced at 1 day, whereas alleviated after 4 days. The was a partial reduction in H2O2-induced POD activity with PMSF and CX, indicating the presence of pre-existing inactive PODs in chloroplasts. We also propose a new role for chloroplastidial proteases as activators of pre-existing inactive PODs during leaf senescence.

Melanogenesis inhibitory, anti-inflammatory, and chemopreventive effects of limonoids from the seeds of Azadirachta indicia A. Juss. (neem).

Thirty-one nortriterpenoids, including 28 limonoids (1-28) and 3 degraded limonoids (29-31), and one diterpenoid (32), were isolated from the seed extract of Azadirachta indica (neem). Among these, six were new compounds and their structures were established to be 15-hydroxyazadiradione (3), 7-benzoyl-17-hydroxynimbocinol (5), 23-deoxyazadironolide (12), limocin E (13), 23-epilimocin E (14), and 7alpha-acetoxy-3-oxoisocopala-1,13-dien-15-oic acid (32). Upon evaluation of compounds 1-32 on the melanogenesis in the B16 melanoma cells, five compounds, 20, 26, 27, 29, and 31, exhibited marked inhibitory effect (74-91% reduction of melanin content at 25 microg/mL) with no or almost no toxicity to the cells. Seven compounds, 1, 6, 9, 10, 18, 20, and 26, on evaluation for their inhibitory effect against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 microg/ear) in mice, exhibited, except for compound 26, marked anti-inflammatory activity (ID(50) values 0.09-0.26 mg/ear). In addition, all of the 32 compounds exhibited moderate or potent inhibitory effects (IC(50) values of 230-501 mol ratio/32 pmol TPA) against the Epstein-Barr virus early antigen (EBV-EA) activation induced by TPA. Furthermore, on evaluation of azadirachtin B (21) for its anti-tumor-initiating activity on the two-stage carcinogenesis of mouse skin tumor induced by peroxynitrite (ONOO-; PN) as an initiator and TPA as a promoter, this exhibited marked inhibitory activity.

Javanicin, an antibacterial naphthaquinone from an endophytic fungus of neem, Chloridium sp.

The endophytic fungus Chloridium sp. produces javanicin under liquid and solid media culture conditions. This highly functionalized naphthaquinone exhibits strong antibacterial activity against Pseudomonas spp., representing pathogens to both humans and plants. The compound was crystallized and the structure was elucidated by X-ray crystallography. The X-ray structure confirms the previously elucidated structure of the compound that was done under standard spectroscopic methods. The importance of javanicin in establishing symbiosis between Chloridium sp. and its host plant, Azadirachta indica, is briefly discussed.

Interspecific variation in vessel size, growth and drought tolerance of broad-leaved trees in semi-arid regions of Kenya.

In semi-arid regions, trees often wither during the dry season. Withering is sometimes manifest as die-back, whereby whithering results in shoot death, which progresses downward from the uppermost part of the crown. In this study, we measured the relationships between height growth and diameter at breast height, die-back frequency and severity, vessel size and specific hydraulic conductivity of four evergreen (Senna siamea (Lamk) H.S. Irwin & Barneby, Jacaranda mimosifolia D. Don, Azadirachta indica A.H.L. Juss and Acacia gerrardii Benth.) and one deciduous (Melia volkensii Gürke) plantation tree species in Kenya, which has a conspicuous dry season. Die-back occurred readily in some species, but not in others. Senna siamea showed the highest specific hydraulic conductivity and the highest growth rate among the five species and was quite susceptible to die-back. Among species, height growth and specific hydraulic conductivity were positively correlated with vessel size and negatively correlated with die-back frequency, suggesting a trade-off between growth rate and drought tolerance. This implies that an adaptation to rapid growth under humid conditions leads to low drought tolerance. However, the deciduous tree Melia volkensii showed high specific hydraulic conductivity and growth, with no symptoms of die-back, implying that a mechanism associated with the deciduous habit results in drought avoidance by reducing the requirement for water during the dry season.

Production of haploids of neem (Azadirachta indica A. Juss.) by anther culture.

Androgenic haploids of the neem tree (Azadirachta indica A. Juss.) were produced by anther culture at the early- to late-uninucleate stage of pollen. Haploid formation occurred via callusing. The best medium for inducing callusing in the anther cultures was Murashige and Skoog's basal medium (MS) (9% sucrose) supplemented with 1 microM 2,4-D, 1 microM NAA and 5 microM BAP, while anther callus multiplied best on MS medium supplemented with 1 microM 2,4-D and 10 microM Kn. These calli differentiated shoots when transferred to a medium containing BAP; 5 microM BAP was optimum for young calli (75% cultures differentiated shoots), but older calli showed the best regeneration with 7.5 microM BAP. Shoots elongated at a lower concentration of BAP-0.5 microM. These shoots were multiplied by forced axillary branching and rooted in vitro. The plants were subsequently established in soil. Of the plants that regenerated from anther callus 60% were haploid, 20% were diploid and 20% were aneuploid.

Studies on the use of neem products for removal of ammonia from brackishwater.

Laboratory experiments were carried out to investigate the ammonia removal from brackishwater (Salinity 16 +/- 1 ppt) using natural plant products such as seed powder and seed oil from neem (Azadirachta indica) and commercially available neem products-neemazal and neemgold. The experimental results showed that ammonia removal was effective with 90 mg/l of neem oil, whereas, neem seed powder at 90 mg/l registered an increase in ammonia levels throughout the course of the experiment. Neem oil, neemazal, and neemgold at 90 mg/l were effective in decreasing the total ammonia nitrogen (TAN) level of 0.40-0.45 mg/l in 96 h. The effect of initial ammonia concentrations on the ammonia removal using neem oil revealed that percentage ammonia removal decreased with an increase in initial ammonia concentration.