PERFORMANCE AND PERSISTENCE OF GREEN FLUORESCENT PROTEIN （gfp） MARKED AZOTOBACTER CHROOCOCCUM IN STERILIZED AND UNSTERILIZED WHEAT RHIZOSPHERIC SOIL

ThepracticaluseofAzotobacterchroococcumasbiofertilizer hasbeenincreasinglyidentifiedinrecentyears.Oftentheefficacy ofthesebacteriaisassessedonthebasisofcropresponseswithout knowingthesurvival,persistenceandcompetitiveabilityofthein troducedstrain.Inoculat…     

1H NMR-based metabolomics of time-dependent responses of Eisenia fetida to sub-lethal phenanthrene exposure

¹H NMR-based metabolomics was used to examine the response of the earthworm Eisenia fetida after exposure to sub-lethal concentrations of phenanthrene over time. Earthworms were exposed to 0.025 mg/cm² of phenanthrene (1/64th of the LC₅₀) via contact tests over four days. Earthworm tissues were extracted using a mixture of chloroform, methanol and water, resulting in polar and non-polar fractions that were analyzed by ¹H NMR after one, two, three and four days. NMR-based metabolomic analyses revealed heightened E. fetida responses with longer phenanthrene exposure times. Amino acids alanine and glutamate, the sugar maltose, the lipids cholesterol and phosphatidylcholine emerged as potential indicators of phenanthrene exposure. The conversion of succinate to fumarate in the Krebs cycle was also interrupted by phenanthrene. Therefore, this study shows that NMR-based metabolomics is a powerful tool for elucidating time-dependent relationships in addition to the mode of toxicity of phenanthrene in earthworm exposure studies.     

Natural variability and correlations in the metabolic profile of healthy Eisenia fetida earthworms observed using ¹H NMR metabolomics

¹H NMR metabolomics can be used to assess the sub-lethal toxicity of contaminants to earthworms by identifying alterations in the metabolic profiles of contaminant- exposed earthworms in contrast to those of healthy (control) individuals. In support of this method this study sought to better characterize the baseline metabolic profile of healthy, mature earthworms of the species, Eisenia fetida, which is recommended for both acute and sub-lethal toxicity testing for soil contaminants. Profiles of D₂O-buffer extracted metabolites were determined using ¹H NMR spectroscopy and both inter-individual metabolic variability and pair-wise metabolic correlations were assessed. The control earthworm extracts exhibited low overall inter-individual metabolic variability, with a spectrum-wide median relative standard deviation (%RSD = standard deviation/mean × 100) of 14%, which suggests that the metabolic profile of E. fetida earthworms is well controlled in laboratory conditions and supports further use of this organism in environmental metabolomics research. In addition, strong positive correlations were detected between the levels of maltose, betaine, glycine, and glutamate as well as between the levels of lactate, valine, leucine, alanine, lysine, tyrosine, and phenylalanine which had not previously been reported. Since comparison of pair-wise metabolic correlations between control and treated organisms can reveal changes in the underlying pattern of biochemical relationships between the metabolites, identification of these significant metabolic correlations in control earthworms provides an additional characteristic that may be applied to delineate between control and treated earthworms in future NMR-based metabolomic studies.     

H NMR-based metabolomics investigation of Daphnia magna responses to sub-lethal exposure to arsenic,copper and lithium

Metal and metalloid contamination constitutes a major concern in aquatic ecosystems. Thus it is important to find rapid and reliable indicators of metal stress to aquatic organisms. In this study, we tested the use of ¹H nuclear magnetic resonance (NMR) – based metabolomics to examine the response of Daphnia magna neonates after a 48 h exposure to sub-lethal concentrations of arsenic (49 μg L⁻¹), copper (12.4 μg L⁻¹) or lithium (1150 μg L⁻¹). Metabolomic responses for all conditions were compared to a control using principal component analysis (PCA) and metabolites that contributed to the variation between the exposures and the control condition were identified and quantified. The PCA showed that copper and lithium exposures result in statistically significant metabolite variations from the control. Contributing to this variation was a number of amino acids such as: phenylalanine, leucine, lysine, glutamine, glycine, alanine, methionine and glutamine as well as the nucleobase uracil and osmolyte glycerophosphocholine. The similarities in metabolome changes suggest that lithium has an analogous mode of toxicity to that of copper, and may be impairing energy production and ionoregulation. The PCA also showed that arsenic exposure resulted in a metabolic shift in comparison to the control population but this change was not statistically significant. However, significant changes in specific metabolites such as alanine and lysine were observed, suggesting that energy metabolism is indeed disrupted. This research demonstrates that ¹H NMR-based metabolomics is a viable platform for discerning metabolomic changes and mode of toxicity of D. magna in response to metal stressors in the environment.     

Fly ash bricks development using concrete waste debris and self-healing bacteria

Journal of Material Cycles and Waste Management - In this study, an effort has been made to use concrete waste debris for the manufacturing of fly ash bricks as a sustainable material. Though in...