Effect of emulsion size and shelf life of azadirachtin A on the bioefficacy of neem (Azadirachta indica A. Juss) emulsifiable concentrates

In a study of 33 recipes of neem oil based emulsifiable concentrates, the specific surface area of the emulsions and cream plus oil layer separation in emulsions at 24 h revealed a correlation of -0.6874 between them and correlations of -0.8940 and 0.6972, respectively, with bioefficacy (LC(50)) against the 3-day-old second-instar larvae of the Bihar hairy caterpillar, Spilosoma obliqua Walker. Nearly 96-99% of azadirachtin A in emulsifiable concentrates (aza-A content = 617.93-1149.65 ppm) degraded during the heat stability test at 54 +/- 1 degrees C for 14 days with half-lives ranging between 1.84 and 4.53 days. The LC(50) values against S. obliqua were, however, statistically at par in both the pre- and the post-heat-treated samples, suggesting a similar effect of azadirachtin A and its degradation products on the bioactivity. The half-life of azadirachtin A could be enhanced by storing the concentrates at lower temperatures. A low pH of the formulation solvent did not check the degradation of azadirachtin A, as reported with aqueous solutions in the literature.     

Response of neem (Azadirachta indica A. Juss) to indigenous arbuscular mycorrhizal fungi, phosphate-solubilizing and asymbiotic nitrogen-fixing bacteria under tropical nursery conditions

Neem (Azadirachta indica A. Juss) seedlings were inoculated with arbuscular mycorrhizal (AM) fungi, Glomus intraradices Schenck and Smith and G. geosporum (Nicol. and Gerd.) Walker, Azospirillum brasilense, and phosphate-solubilizing bacteria (PSB) individually or in various combinations in unsterile soil under nursery conditions. Seedlings were harvested at 60 and 120 days after transplantation. Microbial inoculation resulted in increased mycorrhizal colonization, greater plant height, leaf area and number, root collar diameter, biomass, phosphorus, nitrogen and potassium content, and seedling quality. Inoculated seedlings also had low root/shoot ratios and low nutrient utilization efficiencies. Populations of PSB declined with seedling growth; contrarily populations of A. brasilense increased. A. brasilense and PSB populations were related to each other and influenced root colonization by AM fungi. Microbial inoculation effects were greatest when seedlings were inoculated with a combination of microbes rather than individually. This clearly indicates that these microorganisms act synergistically when inoculated simultaneously, with maximum response being when both AM fungi were coinoculated with A. brasilense and PSB. The results emphasize the importance of microbial inoculations for the production of robust, rapidly growing seedlings in nurseries and illustrate the advantage of inoculating soils of a low microbial population with indigenous microbes.     

Influence of operating parameters on the use of the microwave-assisted process (MAP) for the extraction of azadirachtin-related limonoids from neem (Azadirachta indica) under atmospheric pressure conditions

The use of the microwave-assisted process (MAP) for the extraction of azadirachtin-related limonoids (AZRL) from various parts of the neem tree was investigated under different operating conditions. The influence of microwave power, solvent, and irradiation time on the recovery of AZRL was studied. The efficiency of the microwave-assisted extraction (MAE) of the seed kernel, the seed shell, the leaf, and the leaf stem was compared to that of conventional extraction methods. The content of AZRL in the extracts was estimated with a vanillin-based colorimetric assay and a multivariate calibration technique. The results showed that the MAE technique can enhance the extraction of AZRL from different parts of neem possessing microstructures. Investigation of the influence of the solvent also indicted that the solvent used not only influences the efficiency but also affects the selectivity of the MAE.     

Comparative analysis of seed morphometric and allozyme data among four populations of neem (Azadirachta indica)

The objectives of this study were to determine the patterns of variability and to compare genetic distance between seed morphometric and allozyme data of neem (Azadirachta indica). Phenotypic variation from open pollinated seed materials at 12 isozyme loci was examined using starch gel electrophoresis for four populations from Thailand, Bangladesh and Kenya. All four populations exhibited high levels of variation for seed parameters and isozymes. UPGMA cluster analysis of Nei's genetic distance when based on isozymes revealed two genetic groups. The population from Kenya showed a close genetic relationship with the Bangladeshi populations. The population from Thailand exhibited a distant relationship with the other three populations. This result suggests that the Kenyan population was introduced from the Indian subcontinent. In contrast, cluster analysis of seed morphometric data indicated three different groups of populations. The population from Kenya showed a distant relationship with all other populations. The study demonstrates that allozyme data may not correspond with morphometric traits when measuring genetic distance. It is suggested that this controversy may depend on environmental influence on morphometric traits, possibly also on the enzyme patterns.     

Prenylated flavanones isolated from flowers of Azadirachta indica (the neem tree) as antimutagenic constituents against heterocyclic amines

Four prenylated flavanones were isolated from the methanol extract of the flowers of Azadirachta indica (the neem tree) as potent antimutagens against Trp-P-1 (3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole) in the Salmonella typhimurium TA98 assay by activity-guided fractionation. Spectroscopic properties revealed that those compounds were 5,7,4'-trihydroxy-8-prenylflavanone (1), 5,4'-dihydroxy-7-methoxy-8-prenylflavanone (2), 5,7,4'-trihydroxy-3',8-diprenylflavanone (3), and 5,7,4'-trihydroxy-3',5'-diprenylflavanone (4). All isolated compounds were found for the first time in this plant. The antimutagenic IC(50) values of compounds 1-4 were 2.7 +/- 0.1, 3.7 +/- 0.1, 11.1 +/- 0.1, and 18.6 +/- 0.1 microM in the preincubation mixture, respectively. These compounds also similarly inhibited the mutagenicity of Trp-P-2 (3-amino-1-methyl-5H-pyrido[4,3-b]indole) and PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine). All of the compounds 1-4 strongly inhibited ethoxyresorufin O-dealkylation activity of cytochrome P450 1A isoforms, which catalyze N-hydroxylation of heterocyclic amines. However, compounds 1-4 did not show significant inhibition against the direct-acting mutagen NaN(3). Thus, the antimutagenic effect of compounds 1-4 would be mainly based on the inhibition of the enzymatic activation of heterocyclic amines.     

Variability in Neem (Azadirachta indica) with respect to azadirachtin content

There is a controversy over variations in azadirachtin content in neem (Azadirachta indica) seeds among various provenances and countries. Also, variations in azadirachtins are usually attributed to climatic conditions such as temperature and humidity. The present study was undertaken to evaluate qualitative and quantitative variability in azadirachtins A and B among various neem provenances or individual neem trees. Forty-three provenances of India were examined for intraprovenance variability in azadirachtin A and B content and oil percentage. Twenty-eight individual neem trees from five provenances of different agroclimatic regions were also examined for interprovenance variability. The azadirachtins were quantified using reversed phase analytical HPLC. There were wide variations in oil and azadirachtin contents among different provenances. Azadirachtin A ranged from 556.9 to 3030.8 mg kg(-)(1) of kernels, whereas azadirachtin B was in the range 43.1-590.6 mg kg(-)(1) of kernel among the provenances investigated. Analysis of variance among various neem provenances showed significant differences in oil content, azadirachtin A, total azadirachtin (A + B), and A:B ratio. There were individuals with high and low azadirachtins within a single provenance, and this trend was observed in all of the provenances selected from five agroclimatic regions of the country. Variations among individual trees of a particular provenance indicated that climatic factors such as rainfall, humidity, or temperature did not influence azadirachtin content in the neem trees. The present study shows that there are individual genetic differences among neem trees. A systematic study for tree improvement with a population of mother trees with desired traits should be undertaken by performing half-sib progeny trials and further selections by clonal propagations. The role of genetic makeup needs further research.     

Influence of soil parameters on the effect of 3,4-dimethylpyrazole-phosphate as a nitrification inhibitor

Nitrification inhibitors specifically retard the oxidation of NH₄⁺ to NO₂^- during the nitrification process in soil. In this study, the influence of soil properties on the nitrification-inhibiting effect of 3,4-dimethylpyrazole-phosphate (DMPP), a newly developed nitrification inhibitor, has been investigated. Based on short-term incubation experiments, where the degradation of DMPP could be largely disregarded, the oxidation of the applied NH₄⁺ was more inhibited in sandy soils compared with loamy soils. The influence of soil parameters on the relative NO₂^- formation could be described by a multiple regression model including the sand fraction, soil H⁺ concentration and soil catalase activity (R²=0.62). Adsorption studies showed that the binding behaviour of DMPP was influenced markedly by soil textural properties, viz. the clay fraction (r²=0.61). The adsorption of DMPP was found to be an important factor for the inhibitory effect on NH₄⁺ oxidation in a short-term incubation (r²=0.57). It is concluded that the evaluated soil properties can be used to predict the short-term inhibitory effect of DMPP in different soils. The significance of these results for long-term experiments under laboratory and field conditions needs further investigation.     

Influence of some physicochemical and biological parameters on soil structure formation in alluvial soils

This study examines the role of abiotic (texture, calcium carbonates or iron) and biotic parameters (earthworm and enchytraeid activities) on the initial phases of soil aggregation. Our research focused on humus forms in alluvial soils, which are considered as young and heterogeneous environments. We hypothesized that the soil structure formation is determined by both the nature of the recent alluvial deposits and the soil fauna. For this purpose, six sites were chosen throughout two types of softwood forests (willow and alder forest) representing two stages of vegetation succession. Evidence of soil texture influence on aggregate stability was observed. A dominance of a coarse sand fraction caused a quick colonization of enchytraeids and epigeic earthworms while a silty texture favoured the presence of anecic earthworms, thus increasing the aggregate stabilisation. Iron forms, acting as cementing agents, were observed in the coarse silt, while calcium carbonates were equally distributed among the textural fractions. Active calcium carbonate fraction, binding organic matter with mineral components, was not found in the coarse sand fraction. In conclusion, the tree age cannot alone be used as an indicator of the humus form evolution but biological and physicochemical parameters also influence the initial steps of soil structuration.     

Coating of essential oils onto prilled urea retards its nitrification in soil

Increased use of nitrogenous fertilizers in agriculture has led to the increased pollution of ground water and atmosphere. Certain plant products can be used as coating materials onto urea to reduce the N losses. We evaluated the effectiveness of citronella and palmarosa grass oils as nitrification inhibitors in a soil incubation study. The treatments (14) were combinations of 4 N sources (neem, citronella and palmarosa oil coated prilled ureas, and uncoated prilled urea), 2 coating thicknesses of oils (500 and 1000 mg kg^?1) and 2 N levels (75 and 150 kg N ha^?1), replicated thrice in a randomized block design. N levels at 75 and 150 kg ha^?1 were equivalent to 34 and 68 mg N kg^?1 soil, respectively. Results showed that N sources citronella (CCPU₁₀₀₀) and neem oil (NCPU₁₀₀₀) coated prilled ureas at 1000 mg kg^?1 coating thickness with 75 kg ha^?1 released similar amount of ammonical-N to uncoated prilled urea at 150 kg N ha^?1, suggesting the beneficial effect of coated ureas. The highest nitrification inhibition (%) was recorded with NCPU₁₀₀₀, the reference nitrification inhibitor, which was significantly greater to all the other N sources at 7 days after incubation (DAI), and at par to CCPU₁₀₀₀ at 14 and 21 DAI.     

Influence of moisture regime on inhibition of nitrification by DCD

A laboratory study at the Indian Agricultural Research Institute, New Delhi showed that DCD was an efficient nitrification inhibitor under field capacity moisture (???30 kPa) but not so under submerged soil conditions. Nitrification inhibition by DCD was 79.6% after 1 week of incubation and decreased to 59.3% after 3 weeks of incubation under field capacity moisture, while it was only 19.3% after 1 week of incubation and 10.6% and 18.4% after 2 and 3 weeks of incubation under submergence, respectively. DCD can play an important role in reducing N losses from agricultural fields.     

Influence of smoke exposure on soil enzyme activities and nitrification

Summary Soil was exposed to red phosphorous/butyl rubber (RP/BR) aerosols at various relative humidities in a recirculating environmental wind tunnel. Soil microbial and enzymatic activities were measured immediately after exposure and periodically thereafter for 56 days. The nitrification potential was significantly reduced in soil amended with ammonium sulfate and exposed to RP/BR smoke, and could be related to a decline in soil pH. The rate of nitrate formation in unamended soil with time was also reduced, but by 57 days postexposure, concentrations were similat to those of unexposed controls in all but the thinnest soil lense. Soil dehydrogenase and phosphatase enzyme activities were sensitive to RP/BR smoke and in some treatments no activity was detected. The measured activities did not recover within the 56-day postexposure period and in some cases declined. Soil lense thickness was the greatest factor controlling the degree of RP/BR effects, indicating that injury to soil microbial and enzymatic activities may be surficial. Deposition of smoke particles increased with increasing relative humidity, which had a significant impact on the activities measured.     

Influence of nitrification inhibitors on nitrification and nitrous oxide (N2O) emission from a clay loam soil fertilized with urea

Laboratory incubation experiments were conducted to compare the effects of the nitrification inhibitors 3,4-dimethylpyrazole phosphate (DMPP) and 2-Chloro-6-(trichloromethyl)-pyridine (N-serve) on nitrification and nitrous oxide (N₂O) emission from a Vertosol from southern Australia, under controlled moisture and temperature. Nitrification rates in the control soil were strongly influenced by the temperature and moisture, increasing by a factor of 3.6 for each 10 °C increase between 5 and 25 °C. DMPP inhibited nitrification effectively for 42 days at 5-15 °C and 40-60% water filled pore space (WFPS). DMPP also slowed nitrification appreciably at 25 °C when the soil was at 40% WFPS, but was less effective at 60% water filled pore space. N-serve inhibited nitrification effectively for 42 days under all test conditions. Emissions of N₂O from the urea treatment (no inhibitors) significantly increased with increasing temperature and moisture. The ratio of total N₂O emission to total nitrification was not constant and varied from around 0.03% at 5 °C and 40% WFPS to 0.12% at 25 °C and 60% WFPS. DMPP and N-serve reduced cumulative N₂O emission over 42 days by more than 65% under all the imposed conditions.     

Influence of biocides on tomato nitrogen uptake and soil nitrification and denitirification

As a result of repeated applications, some fungicides may accumulate in the soil to levels high enough to have adverse effects on the activity of soil microorganisms and plant growth. Comparison of the effects of 10 mg kg^‐1 soil of the benlate, captan, and lime‐sulfur fungicides with the nitrification inhibitors (NI) nitrapyrin and terrazole on oxidation of NH₄ ⁺ in Tifton loamy sand (siliceous, thermic plinthic Typic Kandiudults) incubated at 30° C showed that benlate had no significant effects whereas captan inhibited nitrification 21% more than lime‐sulfur, but about 20% less than NI. Application of benlate enhanced NO₃ ^‐ reduction to N₂O and N₂ in liquid medium inoculated with soil whereas 50 and 100 mg L"¹ medium of captan and lime‐sulfur compared favorably with the NI in suppressing NO₃ ^‐ and NO₂ ^‐ reductions, but were less effective than the inhibitors when applied at the low rate of 10 mg L^‐1 medium. In a greenhouse study with tomato (Lycopersicon esculentum Mill. cv. ‘Better Boy'), weekly drench applications of 0.25 mg kg^‐1 soil of the test biocides for four weeks with three NH₄ ⁺‐N: NO₃ ^‐‐N ratios showed that benlate applied with 1: 0 N ratio and lime‐sulfur applied with 0: 1 N ratio restricted significantly the plant growth and N uptake. The largest root: shoot ratios, total plant dry weight, and N uptake were obtained with plants fertilized with 1: 1 N ratio in combination with the biocides.     

Influence of soil factors on the soil enzyme inhibition by Cd

A comparative study was conducted on the toxicity of Cd to alkaline phosphatase activity (ALP) and dehydrogenase activity (DHA) in 18 top soils with contrasting soil properties representative of 14 major soil types in China. Soil pH and carbonate content, soil organic matter, and cation exchange capacity (CEC) largely affected the Cd toxicity on two enzyme activities; with the soil pH having only minor effect on the median ecological dose values based on total Cd concentrations (ED_{50 T}). The values of ED_{50 T}/ED_{50 W} (based on water-soluble Cd content) of alkaline phosphatase and dehydrogenase were strongly influenced by pH and CEC contents, which explained up to 71% of the variation for alkaline phosphatase, 82% of the variation for dehydrogenase, and also were significantly correlated with the parameter K_F derived from Freundlich adsorption isotherms. This study suggests that the values of ED_{50 T}/ED_{50 W} could be useful to evaluate the buffer capacity of soils which protects soil enzymes from harmful effects of heavy metal.     

Influence of diterpenes (colophony and abietic acid) and a triterpene (beta-sitosterol) on net N mineralization,net nitrification,soil respiration,and microbial biomass in birch soil

The aim of this study was to examine the effect of common diterpenes (colophony, abietic acid) and triterpene (beta-sitosterol) on carbon (C) and nitrogen (N) transformations in soil under birch (Betula pendula L.). Samples were taken from the organic layer at two study sites, Kivalo (N-poor soil) and Kerimäki (N-rich soil), and incubated with the above-mentioned terpenes in laboratory conditions. Carbon dioxide evolution (C mineralization), net N mineralization, nitrification, and N and C in microbial biomass were measured. All these terpenes increased C mineralization, but decreased net N mineralization. The potential to decrease net N mineralization depended on amount of terpenes, with a stronger effect at a higher amount. Net nitrification in Kerimäki soil (N-rich soil) decreased but was not completely inhibited by terpenes. Effect of terpenes on soil microbial biomass C and N was not so clear, but they tended to increase both. Our study suggests that higher terpenes can act as a carbon source for soil microbial communities.     

A method of selective inhibition to distinguish between nitrification and denitrification as sources of nitrous oxide in soil

Summary Nitrapyrin and C₂H₂ were evaluated as nitrification inhibitors in soil to determine the relative contributions of denitrification and nitrification to total N₂O production. In laboratory experiments nitrapyrin, or its solvent xylene, stimulated denitrification directly or indirectly and was therefore considered unsuitable. Low partial pressures of C₂H₂ (2.5–5.0 Pa) inhibited nitrification and had only a small effect on denitrification, which made it possible to estimate the contribution of denitrification. The contribution of nitrification was estimated by subtracting the denitrification value from total N₂O production (samples without C₂H₂). The critical C₂H₂ concentrations needed to achieve inhibition of nitrification, without affecting the N₂O reductase in denitrifiers, must be individually determined for each set of experimental conditions.     

Stimulation of heterotrophic nitrification and N2O production,inhibition of autotrophic nitrification in soil by adding readily degradable carbon

Journal of Soils and Sediments - This study aimed to test the hypothesis that readily degradable Carbon (C) has contrasting effect on soil N autotrophic and heterotrophic nitrification, can...     

茶多酚对土壤硝化作用的影响研究

研究了茶多酚对土壤硝化作用和土壤中氨氧化菌与亚硝酸氧化菌数量的影响。结果表明茶多酚能抑制硝化作用,抑制率在8 d内达到31.2%~92.6%。它同时也能抑制氨氧化菌和亚硝酸氧化菌的生长,当加入0.05g茶多酚时,氨氧化菌的数量从650个/g(干土)减少至400个/g(干土),亚硝酸氧化菌的数量从920个/g(干土)减少至661个/g(干土)。当加入茶多酚越多,氨氧化菌和亚硝酸氧化菌的数量减少得越快。因此,茶多酚有可能抑制这两类微生物的生长,从而抑制硝化作用。     

Effect of urease inhibitors on nitrification in soil

Abstract. This paper describes experiments on the inhibitory effects of hydroquinone (HQ), phenylphos-phorodiamidate (PPDA) and N-butyl phosphorothioic triamide (NBPT) on the nitrification in soil. Incubations were carried out at 2/3 field capacity at 25°C of soil samples to which either ammonium or nitrite was added together with inhibitors. Addition of PPDA or NBPT did not influence the oxidation of ammonium. HQ. however, retarded the process significantly, and also the accumulation of nitrite. This was confirmed in experiments whereby added nitrite was followed. Some of the differences could be explained by changes in the soil pH. During incubation the evolution of the total mineral nitrogen was not importantly altered by addition of the inhibitors.