Isolation and Identification of Efficient Malathion-Degrading Bacteria from Deep-Sea Hydrothermal Sediment.

The genetic and metabolic diversity of deep-sea microorganisms play important roles in phosphorus and sulfur cycles in the ocean, distinguishing them from terrestrial counterparts. Malathion is a representative organophosphorus component in herbicides, pesticides, and insecticides and is analogues of neurotoxic agent. Malathion has been one of the best-selling generic organophosphate insecticides from 1980 to 2012. Most of the sprayed malathion has migrated by surface runoff to ocean sinks, and it is highly toxic to aquatic organisms. Hitherto, there is no report on bacterial cultures capable of degrading malathion isolated from deep-sea sediment. In this study, eight bacterial strains, isolated from sediments from deep-sea hydrothermal regions, were identified as malathion degradators. Two of the tested strains, Pseudidiomarina homiensis strain FG2 and Pseudidiomarina sp. strain CB1, can completely degrade an initial concentration of 500 mg/L malathion within 36 h. Since the two strains have abundant carboxylesterases (CEs) genes, malathion monocarboxylic acid (MMC α and MMC ß) and dibasic carboxylic acid were detected as key intermediate metabolites of malathion degradation, and the pathway of malathion degradation between the two strains was identified as a passage from malathion monocarboxylic acid to malathion dicarboxylic acid.

The Dynamics of NO3- and NH4+ Uptake in Duckweed Are Coordinated with the Expression of Major Nitrogen Assimilation Genes.

Duckweed plants play important roles in aquatic ecosystems worldwide. They rapidly accumulate biomass and have potential uses in bioremediation of water polluted by fertilizer runoff or other chemicals. Here we studied the assimilation of two major sources of inorganic nitrogen, nitrate (NO3- ) and ammonium (NH4+), in six duckweed species: Spirodela polyrhiza, Landoltia punctata, Lemna aequinoctialis, Lemna turionifera, Lemna minor, and Wolffia globosa. All six duckweed species preferred NH4+ over NO3- and started using NO3- only when NH4+ was depleted. Using the available genome sequence, we analyzed the molecular structure and expression of eight key nitrogen assimilation genes in S. polyrhiza. The expression of genes encoding nitrate reductase and nitrite reductase increased about 10-fold when NO3- was supplied and decreased when NH4+ was supplied. NO3- and NH4+ induced the glutamine synthetase (GS) genes GS1;2 and the GS2 by 2- to 5-fold, respectively, but repressed GS1;1 and GS1;3. NH4+ and NO3- upregulated the genes encoding ferredoxin- and NADH-dependent glutamate synthases (Fd-GOGAT and NADH-GOGAT). A survey of nitrogen assimilation gene promoters suggested complex regulation, with major roles for NRE-like and GAATC/GATTC cis-elements, TATA-based enhancers, GA/CTn repeats, and G-quadruplex structures. These results will inform efforts to improve bioremediation and nitrogen use efficiency.

Insights into thermal hydrolyzed sludge liquor - Identification of plant-growth-promoting compounds.

This study proposes a new path to utilize thermal hydrolyzed sludge (TH sludge) as fertilizer given high value chemical compounds that can promote plant growth were identified in the liquid fraction of TH sludge (TH liquor). Together with micro- and macro-nutrients released/synthesized during thermal hydrolysis, the feasibility of using TH liquor as organic fertilizer was evaluated. Besides high contents of N, P and K, total free amino acids (FAAs) and plant-growth-promoting FAAs (including glutamic acid, leucine and cystine) also presented in high concentration (4.98-6.48 and 1.12-2.73 g/100 g) in the TH liquor. For the first time, phytohormone compound, indole-3-acetic acid, was observed and the content was the highest in TH liquor with 165 °C treatment (165 °C TH liquor). Meantime, 165 °C TH liquor did not have negative impact on the growth of soil microbes, and this product, instead, demonstrated stimulating effect on the plant growth. These results suggest that 165 °C TH liquor has a great potential to be an organic fertilizer. The remaining solids of TH sludge could be converted to valuable biochar. The holistic approach of using TH liquor as organic fertilizer and producing biochar could realize nearly zero-waste discharge in sludge management.

Sodium azide inhibition of microbial activities and impact on sludge floc destabilization.

Absence of sludge deflocculation under prolonged (24 h or longer) conditions with dissolved oxygen (DO) less than 0.5 mg L⁻1 was recently reported. The reduced aerobic microbial activity, was speculated, had been compensated by the activity of other bacterial (i.e. facultative) communities. To assess such a compensation mechanism and to better evaluate impact of overall microbial activity on the flocculation process, SBR sludge samples were inhibited by using sodium azide under various DO conditions. Sludge deflocculated only in the presence of sodium azide, regardless of DO conditions. This was linked to sodium azide's inhibitory effects on the microbes as indicated by the reduced ammonium and DOC removals. Extracellular potassium level in the mixed liquor of azide spiked samples also indicated simultaneous cell lysis. Fluorescence excitation emission matrix (FEEM) analysis of the extracted bound EPS and fluorescence quenching based interaction studies indicated sodium azide had interacted with the EPS components, and especially with the bound EPS proteins. The impact of such interactions on reduced floc stability needs consideration. This study confirmed the importance of overall microbial activity in the biological flocculation process and the role of bacterial communities, other than the aerobes, in mitigating deflocculation under low DO conditions.

Two-stage microbial conversion of crude glycerol to 1,3-propanediol and polyhydroxyalkanoates after pretreatment.

With increasing demand for biodiesel, crude glycerol as a by-product in biodiesel production has been generated and oversupplied. This study, therefore, explored the pretreatment and a subsequent two-stage microbial system to convert crude glycerol into high value-added products: 1,3-propanediol (1,3-PDO) and polyhydroxyalkanoates (PHAs). After pretreatment, long chain fatty acids (LCFAs) could be effectively removed from crude glycerol to eliminate the inhibition effects on subsequent microbial process. In the anaerobic fermentation, when fed treated crude glycerol increased from 20 g/L to 100 g/L, 1,3-PDO yield decreased from 0.438 g/g to 0.345 g/g and accompanied carboxylic acids shifted from acetate and lactate dominant to lactate overwhelmingly dominant. Meanwhile, the relative abundance of Clostridiales sustained around 50% but Enterobacteriales increased from 19% to 53%. Further fed glycerol increase to 140 g/L resulted in severe substrate inhibition, which could be relieved by intermittent feeding. In aerobic process, glycerol anaerobic digestion effluent (ADE) was fed to the consortium of Bacillus megaterium and Corynebacterium hydrocarbooxydans for selectively consumption of carboxylic acids and residual glycerol from 1,3-PDO to produce PHAs as a secondary high value-added product. The consortium accumulated maximum 8.0 g/L poly (3-hydroxybutyrate) (PHB), and 1,3-PDO purity increased from initial 27.7% to almost 100% when fed with 100 g/L glycerol ADE. Overall, this study provided comprehensive and insightful information on microbial conversion of crude glycerol to high value-added products after pretreatment.

High-strength N-methyl-2-pyrrolidone-containing process wastewater treatment using sequencing batch reactor and membrane bioreactor: A feasibility study.

N-methyl-2-pyrrolidone (NMP) is widely used as a solvent in polymeric membrane fabrication process, its elimination from the process wastewater (normally at a high concentration > 1000 mg/L) prior to discharge is essential because of environmental concern. This study investigated the feasibility of treating high-strength NMP-containing process wastewater in a sequencing batch reactor (SBR; i.e., batch feeding and intermittent aerobic/anoxic condition) and a membrane bioreactor (MBR; i.e., continuous feeding and aeration), respectively. The results showed that the SBR with the acclimated sludge was capable of removing >90% of dissolved organic carbon (DOC) and almost 98% of NMP within 2 h. In contrast, the MBR with the acclimated sludge showed a decreasing NMP removal efficiency from 100% to 40% over 15-day operation. The HPLC and LC-MS/MS analytical results showed that NMP degradation in SBR and MBR could undergo different pathways. This may be attributed to the dissimilar bacterial community compositions in the SBR and MBR as identified by 16s rRNA gene sequencing analysis. Interestingly, the NMP-degrading capability of the activated sludge derived from MBR could be recovered to >98% after they were operated at the SBR mode (batch feeding mode with intermittent aerobic/anoxic condition). This study reveals that SBR is probably a more feasible process to treat high-strength NMP-containing wastewater, but residual NMP metabolites in the SBR effluent need to be post-treated by an oxidation or adsorption process in order to achieve zero-discharge of toxic chemicals.

Progress and development of analytical methods for gibberellins.

Gibberellins, as a group of phytohormones, exhibit a wide variety of bio-functions within plant growth and development, which have been used to increase crop yields. Many analytical procedures, therefore, have been developed for the determination of the types and levels of endogenous and exogenous gibberellins. As plant tissues contain gibberellins in trace amounts (usually at the level of nanogram per gram fresh weight or even lower), the sample pre-treatment steps (extraction, pre-concentration, and purification) for gibberellins are reviewed in details. The primary focus of this comprehensive review is on the various analytical methods designed to meet the requirements for gibberellins analyses in complex matrices with particular emphasis on high-throughput analytical methods, such as gas chromatography, liquid chromatography, and capillary electrophoresis, mostly combined with mass spectrometry. The advantages and drawbacks of the each described analytical method are discussed. The overall aim of this review is to provide a comprehensive and critical view on the different analytical methods nowadays employed to analyze gibberellins in complex sample matrices and their foreseeable trends.