Fungitoxicity of lyophilized and spray-dried garlic extracts

Among the compounds discussed for anti-microbial and anti-fungal use allicin (allylthiosulfinate, diallyl disulfide-S-monoxide), an active ingredient of garlic, has attracted considerable attention. The objective of this study is to determine the antifungal activity of a local garlic ecotype (Voghiera) extracts against different pathogens. Primary screening was carried out by the agar plates technique using ethanol garlic extract at four final concentrations against the following organisms: Alternaria alternata, Aspergillus spp., Colletotrichum acutatum, Didymella bryoniae, Fusarium culmorum, Fusarium avenaceum, Fusarium gramineareum, Gliocladium roseum 47, Pythium splendens, Rhizoctonia solani, Sclerotium rolfsii, Stemphylium vesicarium, Trichoderma longibranchiatum, and Botrytis cinerea. Secondary screening was carried out using a lyophilized and a spray-dried preparation at different concentrations against the organisms selected for the high inhibition garlic effect in the primary screening and compared with the commercial fungicides mancozeb and iprodione. The best results were observed for the spray-dried garlic compound that showed a good fungicidal activity at the concentration of 1.5 g/10 mL while lyophilized garlic at the same concentration exhibithed less inhibition activity against the four fungi analyzed in the second screening.     

Insecticidal activity and fungitoxicity of plant extracts and components of horseradish (Armoracia rusticana) and garlic (Allium sativum)

To avoid environmental pollution and health problems caused by the use of traditional synthetic pesticides, there is a trend to search for naturally occurring toxicants from plants. Among the compounds discussed for anti-fungal and insecticidal activity, the natural extracts from garlic and horseradish have attracted considerable attention. The objective of this study is to determine the insecticidal and anti-fungal activity of Armoracia rusticana and Allium sativum L. extracts against larvae of Aedes albopictus (Skuse) and some pathogenic fungi. For the insecticidal test, horseradish and garlic extracts were prepared from fresh plants (cultivated in Emilia Romagna region) in a solution of ethanol 80 % and the two different solutions were used at different concentrations (for the determination of the lethal dose) against the fourth instar mosquito's larvae. The fungicidal test was carried out by the agar plates technique using garlic and horseradish extracts in a 10 % ethanol solution against the following organisms: Sclerotium rolfsii Sacc., Trichoderma longibrachiatum, Botrytis cinerea Pers., Fusarium oxysporum Schlecht. and Fusarium culmorum (Wm. G. Sm.) Sacc. The first results demonstrated that the horseradish ethanol extracts present only a fungistatic activity against Sclerotium rolfsii Sacc., Fusarium oxysporum Schlecht. and F. culmorum (Wm.G. Sm) Sacc. while garlic extracts at the same concentration provided a good fungicidal activity above all against Botrytis cinerea Pers. and S. rolfsii. A. rusticana and A. sativum preparations showed also an interesting and significant insecticidal activity against larvae of A. albopictus, even if horseradish presented a higher efficacy (LC?? value of 2.34 g/L), approximately two times higher than garlic one (LC?? value of 4.48 g/L).     

Comparison of in vitro activities of amphotericin, clotrimazole, econazole, miconazole, and nystatin against Fusarium oxysporum.

The inhibitory activity of amphotericin B, clotrimazole, econazole, miconazole and nystatin was compared against Fusarium oxysporum f.sp. radicis-cucumerinum. The most efficient antifungal agent against the growth of Fusarium oxysporum was econazole, followed by clotrimazole, miconazole, amphotericin and nystatin. The ED50 and ED90 values were 0.053 and 1.002 ppm for econazole, 0.088 and 1.100 ppm for clotrimazole, 0.173 and 3.210 ppm for miconazole, 0.713 and greater than 48 ppm for amphotericin and 3.860 and 16.702 ppm for nystatin. The ED50 values of nystatin and amphotericin against spore germination of Fusarium oxysporum were determined at 3.1427 ppm and 8.3990 ppm respectively, nystatin was 2.76 times more effective than amphotericin, while no effect was observed after the addition of econazole, clotrimazole and miconazole. The tested azoles were more effective than amphotericin and nystatin on growth inhibition of Fusarium oxysporum but amphotericin and nystatin acted significantly better on spore germination of Fusarium.     

The effect of sodium chloride on the two-step kinetics of the nitrifying process.

Sodium chloride affects the transformation rate of several compounds in bioreactors. Most authors report a decrease in microorganism activity at increasing salt concentrations. In this work, a kinetic model that relates sodium chloride concentration with the rates of each step of the nitrification process is proposed; thus, the effect of sodium chloride concentration (0 to 60 g/L) on the nitritation and nitratation rates was separately studied. To carry out the independent study of each step, a combination of the respirometric method with sodium azide, an inhibitor of the nitratation step, was performed. The dot-blot hybridization technique with 16S rRNA-targeted probes was used to determine the ammonia-oxidizing and nitrite-oxidizing bacterial fraction, then it was possible to relate the culture's function with its biological composition. Rates of both steps were linearly reduced at increasing salt concentrations: the nitratation rate was more affected than the nitritation rate. Simulations carried out in a nitrifying sequencing batch reactor indicate that nitrite might accumulate at high salt concentrations. Sodium chloride exerts a reversible inhibition on ammonia oxidation and nitrite oxidation.     

Limestone Dissolution in Flue Gas Scrubbing: Effect of Sulfite

Batch limestone dissolution experiments were carried out in a pH stat apparatus at 55°C with CO2 sparging and dissolved sulfite. Particle size distribution, utilization, sulfite in solution, limestone type, and the approach to calcite equilibrium were all found to contribute to the limestone reactivity. In the absence of sulfite, limestone dissolution was controlled solely by mass transfer. For a given stone under mass transfer control, film thickness was found to be independent of pH. The dissolution rate in the presence of sulfite was controlled by a combined surface kinetics/mass transfer regime. SEM micrographs supported this conclusion. A surface rate correlation was developed which accounted for observed inhibition by an inverse dependence on calcium sulfite concentration at the limestone surface. While the form of the rate expression was applicable to all stones, the surface rate constant was stone dependent. A computer code which accounted for mass transfer with surface kinetics was tested against experimental observations of four limestone types. Changes in pH and the concentrations of calcium, carbonate, sulfite, sulfate, and adipic acid were accurately modeled.     

Tolerance to agricultural pesticides of strains belonging to four genera of Rhizobiaceae

In order to determine their tolerance to pesticides, 122 strains of rhizobia isolated from different geographical regions, and belonging to the genera Rhizobium, Mesorhizobium, Sinorhizobium and Bradyrhizobium were tested against eight herbicides, four fungicides and five insecticides. Sensitivity to the pesticides was measured by using the filter paper disk method at four concentrations, 0.45, 4.5, 45 and 450 μg per disk. When the pesticides were used at 0.45 μg per disk, a concentration similar to that found when pesticides are applied under field conditions, no inhibition was observed. Strains growth was affected at concentrations of 45 and 450 μg pesticide per disk. These higher concentrations can be encountered when seeds are treated with pesticides. Pesticides tolerance level was correlated to pesticide function, i.e rhizobial strains were more tolerant to insecticides, followed by herbicides and then fungicides. Two fungicides, captan and mancozeb, inhibited the highest number of strains. Only one insecticide, carbaryl, affected the growth of some rhizobial strains. Strains isolated from the arctic (Mesorhizobium spp. and R. leguminosarum bv. viciae), a putative pesticides-free environment, were either less or equally affected by pesticides compared to strains isolated from agricultural regions.     

Assessment of Long-Term Radiological Effects on Plants and Animals from a Deep Geological Repository: No Discernible Impact Detected

This study investigates whether non-human biota are protected against harmful effects of ionizing radiation after a possible future release of radioactive matter from a planned repository for spent nuclear fuel. Radiation dose rates to a broad spectrum of organisms were calculated based on data from sampled organisms and modeled activity concentrations. Calculations were performed with the ERICA Tool, a software program which applies a screening dose-rate value of 10 microgray per hour (μGy h^?1) for all types of organisms. Dose rates below this value are thought to result in minimal risk to the individual or population. All calculated dose rates were below the screening value and below the lowest relevant band of “derived consideration levels” proposed by the International Commission on Radiological Protection. This provides a sound basis for arguing that no individuals or populations of examined species would be harmfully affected by a possible radioactive release from the repository.     

Halofenate and clofibrate inhibition of pyruvate dehydrogenase from Fusarium culmorum

Pyruvate dehydrogenase (E1, E.C. 1.2.4.1) was obtained from Fusarium culmorum by ammonium sulfate precipitation. An eight-fold purification was obtained with a specific activity of 13 K units/mg protein. Both halofenate and clofibrate inhibited the enzyme complex non-competitively. The inhibitory effect of halofenate was greater than that of clofibrate being 42% higher at 20 mM concentration compared to the inhibition by clofibrate at 40 mM concentration. Both compounds disorganized the normal cytoplasmic lipids including the emptying of cells in the mycelium suggesting membrane disruption.     

Rhizoremediation of pentachlorophenol by Sphingobium chlorophenolicum ATCC 39723

Sphingobium chlorophenolicum is well known as a pentachlorophenol (PCP) degrader. The objective of this study was to evaluate PCP degradation in a loamy sandy soil artificially contaminated with PCP using phytoremediation and bioaugmentation. Measurements of PCP concentrations were carried out using high performance liquid chromatography analyses (HPLC). The toxic effect of PCP on plants was studied through the monitoring of weight plant and root length. The biodegradation of PCP by S. chlorophenolicum in soil was assessed with a bioluminescence assay of Escherichia coli HB101 pUCD607. Bacterial analyses were carried out by plating on Mineral Salt Medium (MSM) for S. chlorophenolicum, MSM for PCP-degrading/tolerant organisms and Trypticase Soy Broth Agar (TSBA) for heterotrophic organisms. The introduction of S. chlorophenolicum into soil with plants showed a faster degradation when compared to the non-inoculated soil. The monitoring of the plant growth showed a protective role of S. chlorophenolicum against the toxicity of PCP. The bioassay confirmed that initial toxicity was lowered while degradation progressed. There was a significant increase of organisms tested in the roots in comparison to those in the soil. This study showed that the presence of S. chlorophenolicum enhanced the PCP degradation in a loamy soil and also it had a protective role to prevent phytotoxic effects of PCP on plant growth. The combined use of bioaugmentation and plants suggests that the rhizosphere of certain plant species may be important for facilitating microbial degradation of pesticides in soil with important implications for using vegetation to stabilize and remediate surface soils.     

COMPARISON OF IN VITRO ACTIVITIES OF AMPHOTERICIN,CLOTRIMAZOLE, ECONAZOLE,MICONAZOLE, AND NYSTATIN AGAINST FUSARIUM OXYSPORUM

The inhibitory activity of amphotericin B, clotrimazole, econazole, miconazole and nystatin was compared against Fusarium oxysporum f.sp. radiciscucumerinum. The most efficient antifungal agent against the growth of Fusarium oxysporum was econazole, followed by clotrimazole, miconazole, amphotericin and nystatin. The ED_{5 0} and ED_{9 0} values were 0.053 and 1.002 ppm for econazole, 0.088 and 1.100 ppm for clotrimazole, 0.173 and 3.210 ppm for miconazole, 0.713 and greater than 48 ppm for amphotericin and 3.860 and 16.702 ppm for nystatin. The ED_{5 0} values of nystatin and amphotericin against spore germination of Fusarium oxysporum were determined at 3.1427 ppm and 8.3990 ppm respectively, nystatin was 2.76 times more effective than amphotericin, while no effect was observed after the addition of econazole, clotrimazole and miconazole. The tested azoles were more effective than amphotericin and nystatin on growth inhibition of Fusarium oxysporum but amphotericin and nystatin acted significantly better on spore germination of Fusarium.     

Effect of mycosynthesized silver nanoparticles from filtrate of Trichoderma harzianum against larvae and pupa of dengue vector Aedes aegypti L

Mosquitoes transmit dreadful diseases, causing millions of deaths every year. Therefore, screening for larvicidal and pupicidal activity of microbial extracts attributes could lead to development of new and improved mosquito control methods that are economical and safe for nontarget organisms and are ecofriendly. Synthetic chemical insecticides occupy predominant position in control strategies. These hazardous chemicals exert unwarranted toxicity and lethal effects on nontarget organisms, develop physiological resistance in target, and cause adverse environmental effect. For vector control, fungal-mediated natural products have been a priority in this area at present. In the current study, effective larvicidal and pupicidal effect of mycosynthesized silver nanoparticles (Ag NPs) using an entomopathogenic fungi Trichoderma harzianum against developmental stages of the dengue vector Aedes aegypti was investigated. An attractive possibility of green nanotechnology is to use microorganisms in the synthesis of nanosilver especially Ag NPs. The mycosynthesized Ag NPs were characterized to find their unique properties through UV-visible spectrophotometer, X-ray diffraction analysis, Fourier transform infrared, and surface characteristics by scanning electron microscopy. To analyze the bioefficacy, different test concentrations for extracellular filtrate (0.2, 0.4, 0.6, 0.8, and 1.0 %) and Ag NPs (0.05, 0.10, 0.15, 0.20, and 0.25 %) were prepared to a final volume of 200 mL using deionized water; 20 larvae of each instars (I–IV) and pupa were exposed to each test concentration separately which included a set of control (distilled water) group with five replicates. Characterization of the synthesized Ag NPs were about 10–20 nm without aggregation. Susceptibility of larval instars to synthesized Ag NPs was higher than the extracellular filtrate of T. harzianum alone after 24-h exposure, where the highest mortality was recorded as 92 and 96 % for first and second instars and 100 % for third, fourth instars, and pupa. Lethal concentration 50 values of 0.079, 0.084, 0.087, 0.068, and 0.026 % were recorded for I–IV instars and pupa, respectively, when exposed to Ag NPs at 0.25 % concentration. Toxicity was exhibited against first (1.076 %), second (0.912 %), third (0.770 %), fourth (0.914 %) instars larvae, and pupa (0.387 %) with extracellular filtrate at a concentration of 1 % that was three- to fourfold higher compared to Ag NPs; no mortality was observed in the control. The present study is the first report on effective larvicidal and pupicidal activity of Ag NPs synthesized from an entomopathogenic fungi T. harzianum extracellular filtrate and could be an ideal ecofriendly, single-step, and inexpensive approach for the control of A. aegypti.     

Potentiation of antifungal effect of a mixture of two antifungal fractions obtained from Baccharis glutinosa and Jacquinia macrocarpa plants

The aim of the present work was to evaluate the effect of mixtures of antifungal fractions extracted from Baccharis glutinosa and Jacquinia macrocarpa plants on the development of the filamentous fungi Aspergillus flavus and Fusarium verticillioides. The minimal inhibitory concentration that inhibited 50% of growth (MIC₅₀) of each plant antifungal fraction was determined from the percentage radial growth inhibition of both fungi. Binomial mixtures made with both plant fractions were used at their MIC₅₀ to determine the Fractional Inhibitory Concentration index (FIC index) for each fungus in order to evaluate their synergistic effect. Each synergistic mixture was analyzed in their effect on spore germination, spore size, spore viability, mitotic divisions, hyphal diameter and length, and number of septa per hypha. Some antifungal mixtures, even at low concentrations, showed higher antifungal effect than those of the individual antifungal fraction. The FIC indices of mixtures that showed the highest antifungal activity against A. flavus and F. verticillioides were 0.5272 and 0.4577, respectively, indicating a synergistic effect against both fungi. Only 12% and 8% of the spores of A. flavus and F. verticillioides, respectively, treated with the synergistic mixtures, were able to germinate, although their viability was not affected. An increase in the number of septa per hypha of both fungi was observed. The results indicated that the synergistic mixtures strongly affected the fungal growth even at lower concentrations than those of the individual plant fractions.     

镉胁迫对万寿菊生长及生理生态特征的影响

通过水培实验,研究了不同Cd浓度(0、0.1、0.5、1、2和5 mg/L)胁迫对万寿菊生长及生理生化指标的影响.研究表明,低浓度Cd(＜0.5 mg/L)胁迫下,万寿菊的生长末受到显著影响,说明万寿菊对低浓度Cd有一定的耐性.而Cd浓度超过0.5 mg/L时,万寿菊的相对生长速率较对照明显降低(P＜0.05),最高可...     

Changes in microbiological metabolism under chemical stress

Chemical stress may alter microbiological metabolism and this, in turn, may affect the natural and engineered systems where these organisms function. The impact of chemical stress on microbiological metabolism was investigated using model chemicals 2,4-dinitrophenol (DNP), pentachlorophenol (PCP), and N-ethylmaleimide (NEM). Biological activity of Pseudomonas aeruginosa was measured in batch systems, with and without stressors at sub-lethal concentrations. Stressor DNP, between 49 and 140 mg l(-1), and PCP, at 15 and 38 mg l(-1), caused decreases in biomass growth yields, but did not inhibit substrate utilization rates. These effects increased with stressor concentrations, showing as much as a 10% yield reduction at the highest DNP concentration. This suggests that a portion of carbon and energy resources are diverted from growth and used in stress management and protection. Stressor DNP, between 300 and 700 mg l(-1), and PCP at 85 mg l(-1) caused decreases in growth yields and substrate utilization rates. This suggests an inhibition of both anabolism and catabolism. Stressor NEM was the most potent, inhibiting biological activity at concentrations as low as 2.7 mg l(-1). These findings will ultimately be useful in better monitoring and management of biological treatment operations and contaminated natural systems.     

Acute, chronic and sublethal effects of the herbicide propanil on Daphnia magna

Acute and chronic toxicity tests with propanil were conducted on Daphnia magna. The 24 and 48 h LC50 were 43.74 and 5.01 mg/l respectively. Chronic toxicity tests were carried out using sublethal propanil concentrations (0.07, 0.10, 0.21 and 0.55 mg/l) during 21 days. The effect of propanil on survival, reproduction and growth of D. magna organisms was monitored. The parameters used to evaluate herbicide effect on reproduction were: mean total young ones per female, mean brood size, time to first reproduction, mean number broods per female and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to the herbicide. Reproduction was significantly reduced when propanil concentration increased in the medium. The intrinsic rate of natural increase (r) decreased with increasing concentrations of propanil especially in those animals exposed to 0.55 mg/l. However, growth as well as survival of the exposed organisms only decreased in daphnids exposed to the highest propanil concentration tested. The maximum acceptable toxicant concentration (MATC) was calculated for D. magna exposed to the herbicide using as parameter of evaluation the intrinsic rate of natural increase (r). The interpolation of these results gave MATC values of 0.08 mg/l herbicide. We have derived the EC50 values for some selected parameters on D. magna exposed to propanil. EC50 values indicated that reproductive parameters were very sensitive of the effect of propanil on daphnids. Finally, the daphnids were exposed to the same sublethal herbicide concentrations as in the chronic study and the effect of the toxicant on filtration and ingestion rates was determined. Feeding rates of D. magna declined with increasing propanil concentrations. The effective propanil concentrations at which feeding rates were reduced to 50% of that in controls (EC50) were also calculated.     

Equilibrium partitioning theory to predict the sediment toxicity of the anionic surfactant C(12)-2-LAS to Corophium volutator

The study of the effect of the sorption of linear alkylbenzene sulfonates (LAS) on the bioavailability to marine benthic organisms is essential to refine the environmental risk assessment of these compounds. According to the equilibrium partitioning theory (EqP), the effect concentration in water-only exposure will be similar to the effect concentration in the sediment pore water. In this work, sorption and desorption experiments with two marine sediments were carried out using the compound C₁₂-2-LAS. The effect of the sediment sorption on the toxicity of benthic organisms was studied in water-only and in sediment bioassays with the marine mud shrimp Corophium volutator. In addition, three common spiking methods were tested for its application in the toxicity tests, as well as the stability of the surfactant during the water-only and sediment-water test duration. LC50 values obtained from water-only exposure showed a good correspondence with the pore water concentrations calculated from the sorption and desorption isotherms in the spiked sediments.     

高氯离子味精尾母液废水厌氧处理研究

味精尾母液废水COD浓度极高同时含有大最氯离子.采用UASB反应器对味精尾母液废水进行处理,其中接种污泥来自啤酒厂UASB反应器.实验表明:当氯离子浓度在4 500 mg/L以下时,对厌氧微生物没有明礁的抑制作用;5 000 mg/L的氯离子浓度可以看作一个抑制限值,但经过驯化后,仍可获得较好的COD去除效果;当氯离子浓度达到8 000mg/L左右时,COD平均去除率在80%以上.     

Inhibition of laccase activity from Trametes versicolor by heavy metals and organic compounds

Due to the numerous biotechnological applications of laccase enzyme, it is essential to know the influence of different agents usually present in the natural environment on its enzymatic action, especially for in situ treatment technologies. In the present work, a simple and rapid method to determine the inhibitory or inducer effect of different compounds on laccase activity was developed. The compounds tested were copper-chelating agents and heavy metals. It was found that using syringaldazine as a substrate, all copper-chelating agents (except EDTA) highly inhibited laccase activity (around 100%) at an inhibitor concentration lower than 20 mM. Moreover, 40% of inhibition, which was detected at a concentration of 20 mM for both Cd(2+) and Cu(2+) increased with concentration until nearly complete inhibition at 80 mM.