Microbial Necromass, Lignin, and Glycoproteins for Determining and Optimizing Blue Carbon Formation.

Coastal wetlands contribute to the mitigation of climate change through the sequestration of "blue carbon". Microbial necromass, lignin, and glycoproteins (i.e., glomalin-related soil proteins (GRSP)), as important components of soil organic carbon (SOC), are sensitive to environmental change. However, their contributions to blue carbon formation and the underlying factors remain largely unresolved. To address this paucity of knowledge, we investigated their contributions to blue carbon formation along a salinity gradient in coastal marshes. Our results revealed decreasing contributions of microbial necromass and lignin to blue carbon as the salinity increased, while GRSP showed an opposite trend. Using random forest models, we showed that their contributions to SOC were dependent on microbial biomass and resource stoichiometry. In N-limited saline soils, contributions of microbial necromass to SOC decreased due to increased N-acquisition enzyme activity. Decreases in lignin contributions were linked to reduced mineral protection offered by short-range-ordered Fe (FeSRO). Partial least-squares path modeling (PLS-PM) further indicated that GRSP could increase microbial necromass and lignin formation by enhancing mineral protection. Our findings have implications for improving the accumulation of refractory and mineral-bound organic matter in coastal wetlands, considering the current scenario of heightened nutrient discharge and sea-level rise.

In Vitro Anti-Inflammatory Activity of Essential Oil and ß-Bisabolol Derived from Cotton Gin Trash.

Natural α-bisabolol has been widely used in cosmetics and is sourced mainly from the stems of Candeia trees that have become endangered due to over exploitation. The in vitro anti-inflammatory activity of cotton gin trash (CGT) essential oil and the major terpenoid (ß-bisabolol) purified from the oil were investigated against lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages as well as the 3t3 and HS27 fibroblast cell lines. Nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and interleukin 8 (IL-8) were measured using Greiss reagent, enzyme-linked immunosorbent assay (ELISA), and cytokine bead array (CBA)-flow cytometry. Non-toxic concentrations of CGT oil and ß-bisabolol (1.6-50.0 µg/mL) significantly inhibited the production of the inflammatory mediators in a dose-dependent manner. Maximal inhibition by ß-bisabolol was 55.5% for NO, 62.3% for PGE2, and 45.3% for TNF-α production in RAW cells. ß-Bisabolol induced a level of inhibition similar to an equal concentration of α-bisabolol (50.0 µg/mL), a known anti-inflammatory agent. These results suggest ß-bisabolol exerts similar in vitro effects to known topical anti-inflammatory agents and could therefore be exploited for cosmetic and therapeutic uses. This is the first study to report the in vitro anti-inflammatory activity of ß-bisabolol in CGT essential oil.

Isolation and Characterization of Endomycorrhizal Fungi Associated with Growth Promotion of Blueberry Plants.

Despite their notable root mutualism with blueberries (Vaccinium spp.), studies related to Ericoid mycorrhizal (ERM) are relatively limited. In this study, we report the isolation of 14 endomycorrhizal fungi and their identification by fungal colony morphology characterization combined with PCR-amplified fungal internal transcribed spacer (ITS) sequence analyses. Six of the isolated strains were confirmed as beneficial mycorrhizal fungi for blueberry plants following inoculation. We observed the formation of typical ERM hyphae coil structures-which promote and nutritionally support growth-in blueberry seedlings and significant nitrogen and phosphorous content increases in diverse tissues. QRT-PCRs confirmed changes in VcPHT1s expression patterns. After the formation of ERM, PHT1-1 transcription in roots was upregulated by 1.4- to threefold, whilst expression of PHT1-3 and PHT1-4 in roots were downregulated 72% and 60%, respectively. Amino acid sequence analysis of all four VcPHT1s genes from the blueberry variety "Sharpblue" revealed an overall structural similarity of 67% and predicted transmembrane domains. Cloning and overexpression of PHT1-1 and PHT1-3 genes in transgenic Arabidopsis thaliana plants significantly enriched total phosphorus and chlorophyll content, confirming that PHT1-1 and PHT1-3 gene functions are associated with the transport and absorption of phosphorus.

Chemical volatiles present in cotton gin trash: A by-product of cotton processing.

Cotton gin trash (CGT), a waste product of cotton gins, make up about 10% of each bale of cotton bolls ginned. The current study investigates high value volatile compounds in CGT to add value to this by-product. The volatile compounds in CGT and different parts of the cotton plant were extracted using various methods, identified by gas chromatography-mass spectrometry (GC-MS) or nuclear magnetic resonance (NMR) spectroscopy, and then quantified by gas chromatography-flame ionisation detection (GC-FID) against available standards. Terpenoids including monoterpenoids and sesquiterpenoids were found to be the most abundant, making up 64.66% (area under peak) of total volatiles extracted by hydro-distillation. The major extractable terpenoids in CGT were α-pinene (13.69-23.05 µg/g), ß-caryophyllene (3.99-74.32 µg/g), α-humulene (2.00-25.71 µg/g), caryophyllene oxide (41.50-102.08 µg/g) and ß-bisabolol (40.05-137.32 µg/g). Recoveries varied between different extraction methods. The terpenoids were found to be more abundant in the calyx (659.12 µg/g) and leaves (627.72 µg/g) than in stalks (112.97 µg/g) and stems (24.24 µg/g) of the cotton plant, indicating the possible biological origin of CGT volatiles. This study is the first to identify and quantify the different terpenoids present in CGT and significantly, ß-bisabolol, an abundant compound (sesquiterpene alcohol) which may have valuable biological prospects. These findings therefore contribute to identifying alternative management strategies and uses of CGT.

Bioethanol potential of Eucalyptus obliqua sawdust using gamma-valerolactone fractionation.

Optimisation of conditions for gamma-valerolactone (GVL) pretreatment of Australian eucalyptus sawdust for high cellulose biomass and bioethanol production was demonstrated. Pretreatment parameters investigated included GVL concentrations of 35-50% w/w, temperatures of 120-180 °C and reaction durations of 0.5-2.0 h. Optimum conditions were determined using the response surface method (RSM) and central composite face-centred design. Cellulose content increased from 39.9% to a maximum of 89.3% w/w using treatments with 50% GVL at 156 °C for 0.5 h. Temperature had the most significant effect (RSM p < .05) on cellulose content of residual biomass and reducing operational duration of < 0.5 h may be viable according to RSM. PSSF fermentations of optimised pretreated eucalyptus sawdust produced up to 94% theoretical ethanol yield, which corresponded to approximately 181 kg of ethanol per dry ton of eucalyptus sawdust. The compositions of both the residual biomass and pretreatment liquors show that GVL pretreatment is a promising solvent for lignocellulosic biorefining.

Biological Importance of Cotton By-Products Relative to Chemical Constituents of the Cotton Plant.

Although cultivated for over 7000 years, mainly for production of cotton fibre, the cotton plant has not been fully explored for potential uses of its other parts. Despite cotton containing many important chemical compounds, limited understanding of its phytochemical composition still exists. In order to add value to waste products of the cotton industry, such as cotton gin trash, this review focuses on phytochemicals associated with different parts of cotton plants and their biological activities. Three major classes of compounds and some primary metabolites have been previously identified in the plant. Among these compounds, most terpenoids and their derivatives (51), fatty acids (four), and phenolics (six), were found in the leaves, bolls, stalks, and stems. Biological activities, such as anti-microbial and anti-inflammatory activities, are associated with some of these phytochemicals. For example, ß-bisabolol, a sesquiterpenoid enriched in the flowers of cotton plants, may have anti-inflammatory product application. Considering the abundance of biologically active compounds in the cotton plant, there is scope to develop a novel process within the current cotton fibre production system to separate these valuable phytochemicals, developing them into potentially high-value products. This scenario may present the cotton processing industry with an innovative pathway towards a waste-to-profit solution.

Novel Applications for Oxalate-Phosphate-Amine Metal-Organic-Frameworks (OPA-MOFs): Can an Iron-Based OPA-MOF Be Used as Slow-Release Fertilizer?

A porous iron-based oxalate-phosphate-amine metal-organic framework material (OPA-MOF) was investigated as a microbially-induced slow-release nitrogen (N) and phosphorus (P) fertilizer. Seedling growth, grain yields, nutrient uptake of wheat plants, and soil dynamics in incubated soil, were investigated using OPA-MOF vs standard P (triple-superphosphate) and N (urea) fertilizers in an acidic Ferralsol at two application rates (equivalent 120 and 40 kg N ha(-1)). While urea hydrolysis in the OPA-MOF treatment was rapid, conversion of ammonium to nitrate was significantly inhibited compared to urea treatment. Reduced wheat growth in OPA-MOF treatments was not caused by N-deficiency, but by limited P-bioavailability. Two likely reasons were slow P-mobilisation from the OPA-MOF or rapid P-binding in the acid soil. P-uptake and yield in OPA-MOF treatments were significantly higher than in nil-P controls, but significantly lower than in conventionally-fertilised plants. OPA-MOF showed potential as enhanced efficiency N fertilizer. However, as P-bioavailability was insufficient to meet plant demands, further work should determine if P-availability may be enhanced in alkaline soils, or whether central ions other than Fe, forming the inorganic metal-P framework in the MOF, may act as a more effective P-source in acid soils.

Nutrient removal and microbial communities' development in a young unplanted constructed wetland using Bauxsol™ pellets to treat wastewater.

Municipal wastewater was treated over a six month period in an unplanted constructed wetland with a lower soil layer and an upper Bauxsol™ pellet layer. The interactions between Bauxsol™ pellets, soil, effluent and microbial communities demonstrated a positive influence on contaminant removal. Bauxsol™ treated effluent showed >95% phosphate removal and ~26% nitrogen removal during the trial. Substantial quantities of nitrate, trace-metals and Colwell P were bound to the pellets, whereas only ammonium was bound to the soil. The structure of microbial communities analysed by denaturing gradient gel electrophoresis (DGGE) showed distinct bacterial communities attached to Bauxsol™ pellets and soil owing to differences in geochemistry and micro-environmental conditions. Polymerase chain reaction (PCR) amplification of specific marker genes (i.e. bacterial and archaeal amoA genes, nosZ gene, and hzo gene) was used to evaluate the presence of microbial communities associated with nitrogen transformation. Data revealed the co-existence of aerobic ammonia-oxidising bacteria, anaerobic ammonia-oxidising bacteria (anammox) and denitrifiers attached to Bauxsol™ pellets and ammonia-oxidising bacteria and archaea attached to soil. This study successfully demonstrates that Bauxsol™ pellets are a suited alternative media for constructed wetland to treat wastewater effectively removing phosphate and serving as biomass support particles for bacterial communities associated with nitrogen-cycling.

Diversity of microbial communities in an attached-growth system using Bauxsol™ pellets for wastewater treatment.

Columns of Bauxsol™ pellets were used in a field experiment as biomass support particle for wastewater microbial communities. The attached microbial community structure was analysed using denaturing gradient gel electrophoresis (DGGE), targeting the 16S rDNA gene's V3 region. DGGE profiles showed that the type and composition of support particles used (i.e. Bauxsol™ pellets or gravel) had a significant impact on the attached bacterial communities (64% dissimilarity). In addition, ecological indices revealed a more heterogeneous bacterial community structure on the Bauxsol™ pellets. TOC/TN ratios post-experiment (6.5-9.3) suggested a good level of biological activity (i.e. active biofilm) in the Bauxsol™ columns. Moreover, Bauxsol™ pellets were mostly made of inorganic carbon, suggesting insoluble carbonate biomineralisation. Polymerase chain reaction (PCR) amplification of specific marker genes (i.e. bacterial and archaeal amoA genes, nosZ gene, and hzo gene) were used to identify the presence of attached bacterial communities associated with nitrogen transformation. The results along with geochemical data (i.e. up to 50% nitrogen removal) revealed co-existence of ammonia-oxidising bacteria, denitrifiers, and anammox organisms. This study conclusively demonstrates that microbial communities are well-adapted to Bauxsol™ pellets and bacterial communities involved in the nitrogen cycle are present.

Nutrient and trace-metal removal by Bauxsol pellets in wastewater treatment.

In this study, Bauxsol pellets packed in PVC columns were used to remove nutrients and trace-metals from municipal wastewater during a 6 months field trial. Bauxsol pellet columns showed a high phosphate removal rate via precipitation of PO(4)(3-) with Ca(2+) and Mg(2+) ions: at 90% in the 1st month; at 80% from the second to fifth months; and at 60% in the sixth month. Pellet bound total phosphorus and Colwell phosphorus were 7.3 g/kg and 2 g/kg and are about 20 times the concentrations found in most fertile soils. Trace-metals in effluents were bound, probably irreversibly under the columns' environmental conditions, to the Bauxsol minerals that have high surface area to volume ratios and high charge to mass ratios. Experimental results showed a complex nitrogen cycle operating within the Bauxsol pellet columns including anoxic nitrification, denitrification, and anammox processes. Although a transient pH spike, associated with the release of unreacted CaO from the cement binder used in the pellets, was observed, this may be readily corrected through post-treatment pH adjustment. Hence, the geochemistry of Bauxsol pellets can effectively remove and bind nutrients and trace-metals during wastewater treatment, and further research may show that saturated spent pellets can be used as fertilizer.

Minimising alkalinity and pH spikes from Portland cement-bound Bauxsol (seawater-neutralized red mud) pellets for pH circum-neutral waters.

Bauxsol reagents (powder, slurry, or pellet forms) are powerful tools in environmental remediation and water and sewage treatment However, when used in circum-neutral water treatments, cement-bound Bauxsol pellets produce a sustained pH and alkalinity spike due to the presence of unreacted CaO in the cement binder. This study developed a pellet treatment system to minimize the alkalinity/pH spike. The recipe for pelletization consisted of Bauxsol powder, ordinary Portland cement (OPC), hydrophilic fumed silica, aluminum powder, a viscosity modifier, and water. Several batches (including different ratios and sizes) were run using modified makeup waters (H(2)0 + CO(2) or NaHCO(3)) or curing brines (CO(2), NaHCO(3), or Mg/CaCl(2)). Alkalinity, pH stability, and slake durability tests were performed on pellets before and/or after curing. The best result for reducing the alkalinity/pH spike was obtained from a MgCl(2), CaCl(2) bath treatment using a Bauxsol:cement ratio of 2.8:1 (pH 8.28; alkalinity 75.1 mg/L) for a 100 g batch or 245:1 (pH 8.05; alkalinity 35.4 mg/L) for a 1 kg batch. Although brine curing does provide a control on pH/alkalinity release, the pellets may still contain unreacted CaO. Therefore, a freshwater rinse of pellets before treating circum-neutral waters is recommended as is the continued investigation of alternative pellet binders.

Amplification of soil fungal community DNA using the ITS86F and ITS4 primers.

Internal transcribed spacer (ITS) 86F and ITS4 and the ITS1-F and ITS86R primer pairs were tested to specifically amplify fungal community DNA extracted from soil. Libraries were constructed from PCR-amplified fragments, sequenced and compared against sequences deposited in GenBank. The results confirmed that the ITS86F and ITS4 primer pair was selectively specific for the Ascomycetes, Basidiomycetes and Zygomycetes fungal clades. Amplified products generated by the ITS1F and ITS86R primer pair also aligned with sequences from a range of species within the Ascomycete and Basidiomycete clades but not from the Zygomycete. Both primer sets demonstrated fungal specificity and appear to be well suited for rapid PCR-based (fingerprinting) analysis of environmental fungal community DNA. This is the first reported use and assessment of the ITS86F and ITS4 and the ITS1-F and ITS86R primer pairs in amplifying fungal community DNA from soil.

Microbial degradation of the organophosphate pesticide, Ethion.

The organophosphate pesticide, Ethion, remains a major environmental contaminant in rural Australia and poses a significant threat to environmental and public health. The aerobic degradation of Ethion by mesophilic bacteria isolated from contaminated soils surrounding disused cattle dip sites was investigated. Two isolates, identified as Pseudomonas and Azospirillum species, were capable of biodegrading Ethion when cultivated in minimal salts medium. The abiotic hydrolytic degradation products of Ethion such as Ethion Dioxon and O,O-diethylthiosphosphate were not detected. The data suggest the rapid degradation of Ethion to support microbial growth. The results have implications for the development of a bioremediation strategy.

Cloning vectors for Streptococcus thermophilus derived from a native plasmid.

A 3.5-kb native plasmid (pND103) was identified in Streptococcus thermophilus ST2-1. Preliminary sequence analysis indicated that pND103 belongs to group I S. thermophilus plasmids. A region of approximately 2 kb appears to contain three components: a plus origin of replication (ori) typical of plasmids that replicate via rolling circle replication; a gene encoding a replication protein (rep); and a gene encoding a small heat shock protein (hsp). pND103 was then used to construct S. thermophilus/Escherichia coli hybrid cloning vectors by ligating different portions of pND103 to an origin-probe vector (pND330) composed of pUC19 and a Gram-positive erythromycin resistance gene. The shuttle vectors (pND913, pND914 and pND915) were successfully introduced back into plasmid-free S. thermophilus ST3-1 as well as to Lactococcus lactis LM0230 and E. coli JM109. Segregational and structural stability study indicated that these vectors can be maintained in these hosts. The results indicated that pND913, pND914 and pND915 are potential shuttle cloning vectors for S. thermophilus.