Antioxidant and Antimicrobial Properties of Fruiting Body and Submerged Mycelium of Medicinal Mushroom Phellinus robiniae (Agaricomycetes).

This study was conducted to evaluate extraction yield, antioxidant content, antioxidant capacity and antibacterial activity of extracts obtained from submerged mycelium (ME) and fruiting body (FBE) of Phellinus robiniae NTH-PR1. The results showed that yields of ME and FBE reached 14.84 ± 0.63 and 18.89 ± 0.86%, respectively. TPSC, TPC, and TFC were present in both mycelium and fruiting body, and the more contents of them were found in fruiting body. The concentrations of TPSC, TPC and TFC in ME and FBE were 17.61 ± 0.67 and 21.56 ± 0.89 mg GE g-1, 9.31 ± 0.45 and 12.14 ± 0.56 mg QAE g-1, and 8.91 ± 0.53 and 9.04 ± 0.74 mg QE g-1, respectively. EC50 values for DPPH radical scavenging revealed FBE (260.62 ± 3.33 µg mL-1) was more effective than ME (298.21 ± 3.61 µg mL-1). EC50 values for ferrous ion chelating in ME and FBE were 411.87 ± 7.27 and 432.39 ± 2.23 µg mL-1, respectively. Thus, both extracts were able to inhibit Gram-positive and Gram-negative pathogenic bacterial strains, at concentrations ranging in 25-100 mg mL-1 of ME and 18.75-75 mg mL-1 of FBE for Gram-positive bacteria; ranging in 75-100 mg mL-1 of ME and 50-75 of FBE for Gram-negative bacteria. Overall submerged mycelial biomass and fruiting bodies of Ph. robiniae NTH-PR1 can be considered as useful natural sources for development of functional food, pharmaceuticals and cosmetic products or cosmeceuticals.

Insights into the biodegradation of diesel oil and changes in bacterial communities in diesel-contaminated soil as a consequence of various soil amendments.

Soil amendment is a promising strategy to enhance biodegradation capacity of indigenous bacteria. To assess the consequences of various soil amendments before large-scale implementation, a microcosm study was employed to investigate the effects of nutrients (TN), surfactants (TS), oxidants (TO), biochar (TB), and zero-valent iron nanoparticles (nZVI; TNP) on diesel degradation, bacterial communities, and community-level physiological profiles (CLPPs) of legacy field contaminated soil. The results showed that the TN, TB, TNP, TS, and TO, reduced 75.8%, 63.9%, 62.8%, 49.3%, and 40.1% of total petroleum hydrocarbons (TPH), respectively, within 120 days, while control (TW) reduced only 33.8%. In all soil amendments, TPH reduction was positively correlated with oxidation-reduction potential and heterotrophic and TPH-degrading bacteria, while negatively correlated with total nitrogen and available phosphate. Furthermore, in TW, TB, and TNP microcosms, TPH reduction showed positive association with pH, whereas in TN, TS, and TO, TPH reduction was negatively associated with pH. The bacterial diversity was reduced in all treatments as a function of the soil amendment and remediation time: the enriched potential TPH-degrading bacteria were Dyella, Paraburkholderia, Clavibacter, Arthrobacter, Rhodanobacter, Methylobacterium, and Pandoraea. The average well colour development (AWCD) values in CLPPs were higher in TB, sustained and improved in TN, and markedly lower in TNP, TS, and TO microcosms. Overall, these data demonstrate that nutrients and biochar amendments may be helpful in boosting biodegradation, increasing diesel-degrading bacteria, and improving soil physiological functions. In conclusion, diesel degradation efficiency and bacterial communities are widely affected by both type and duration of soil amendments.

Effect of consortium bioaugmentation and biostimulation on remediation efficiency and bacterial diversity of diesel-contaminated aged soil.

This study aimed to evaluate the effects of consortium bioaugmentation (CB) and various biostimulation options on the remediation efficiency and bacterial diversity of diesel-contaminated aged soil. The bacterial consortium was prepared using strains D-46, D-99, D134-1, MSM-2-10-13, and Oil-4, isolated from oil-contaminated soil. The effects of CB and biostimulation were evaluated in various soil microcosms: CT (water), T1 (CB only), T2 (CB + NH4NO3 and KH2PO4, nutrients), T3 (CB + activated charcoal, AC), T4 (CB + nutrients + AC), T5 (AC + water), T6 (CB + nutrients + zero-valent iron nanoparticles, nZVI), T7 (CB + nutrients + AC + nZVI), T8 (CB + activated peroxidase, oxidant), T9 (AC + nZVI), and T10 (CB + nZVI + AC + oxidant). Preliminary evaluation of the bacterial consortium revealed 81.9% diesel degradation in liquid media. After 60 days of treatment, T6 demonstrated the highest total petroleum hydrocarbon (TPH) degradation (99.0%), followed by T1 (97.4%), T2 (97.9%), T4 (96.0%), T7 (96.0%), T8 (94.8%), T3 (93.6%), and T10 (86.2%). The lowest TPH degradation was found in T5 (24.2%), T9 (17.2%), and CT (11.7%). Application of CB and biostimulation to the soil microcosms decreased bacterial diversity, leading to selective enrichment of bacterial communities. T2, T6, and T10 contained Firmicutes (50.06%), Proteobacteria (64.69%), and Actinobacteria (54.36%) as the predominant phyla, respectively. The initial soil exhibited the lowest metabolic activity, which improved after treatment. The study results indicated that biostimulation alone is inadequate for remediation of contaminated soil that lacks indigenous oil degraders, suggesting the need for a holistic approach that includes both CB and biostimulation. Graphical Abstract.

Oil-degrading properties of a psychrotolerant bacterial strain, Rhodococcus sp. Y2-2, in liquid and soil media.

The aim of this study was to investigate oil-degrading ability of newly isolated strain Rhodococcus Y2-2 at low temperature. Rhodococcus sp. Y2-2 was isolated from oil-contaminated soil sampled at the end of winter using a newly developed transwell plate method. In the liquid phase, the oil-degradation efficiency of strain Rhodococcus sp. Y2-2 was about 84% with an initial concentration of 1500 ppm TPH (500 ppm each of kerosene, gasoline, and diesel) when incubated for 2 weeks under optimal conditions: 10 °C, pH 7, and 0.5 g L- 1 inoculum. In the soil phase, the isolate showed 80% oil degradation efficiency using glucose as a carbon source, with an initial concentration of 4000 ppm TPH and the addition of water during 14 days of incubation at 10 °C. Additionally, the degradation efficiency of the isolate was increased by the addition of mixture of surfactant alpha olefin sulfonate and gelatin, although strain Y2-2 also produced many biosurfactant components. This study shows Rhodococcus sp. Y2-2 can degrade oil components both in liquid and soil media by consuming kerosene, gasoline, and diesel as a carbon and energy source. Therefore, the crude oil-degrading ability of Rhodococcus sp. Y2-2 at low temperature provides proper bioremediation tool to clean up oil-contaminated sites especially in cold area or during winter season.

Mesorhizobium soli sp. nov., a novel species isolated from the rhizosphere of Robinia pseudoacacia L. in South Korea by using a modified culture method.

Strain NHI-8(T) was isolated from a forest soil sample, collected in South Korea, by using a modified culture method. Comparative analysis of its nearly full-length 16S rRNA gene sequence showed that strain NHI-8(T) belongs to the genus Mesorhizobium and to be closely related to Mesorhizobium chacoense PR5(T) (97.32 %). The levels of DNA-DNA relatedness between strain NHI-8(T) and reference type strains of the genus Mesorhizobium were 32.28-53.65 %. SDS-PAGE of total soluble proteins and the sequences of the housekeeping genes recA, glnII, and atpD were also used to support the clade grouping in rhizobia. The new strain contained summed feature 8 (57.0 %), cyclo-C19:0ω8c (17.3 %), and C18:0 (11.0 %) as the major fatty acids, as in genus Mesorhizobium. The strain contained cardiolipin, phosphatidylglycerol, ornithine-containing lipid, phosphatidylethanolamine, phosphatidyl-N-dimethylethanolamine, and phosphatidylcholine. Morphological and physiological analyses were performed to compare the characteristics of our strain with those of the reference type strains. Based on the results, strain NHI-8(T) was determined to represent a novel member of the genus Mesorhizobium, and the name Mesorhizobium soli is proposed. The type strain is NHI-8(T) (=KEMB 9005-153(T) = KACC 17916(T) = JCM 19897(T)).

Simple surface foam application enhances bioremediation of oil-contaminated soil in cold conditions.

Landfarming of oil-contaminated soil is ineffective at low temperatures, because the number and activity of micro-organisms declines. This study presents a simple and versatile technique for bioremediation of diesel-contaminated soil, which involves spraying foam on the soil surface without additional works such as tilling, or supply of water and air. Surfactant foam containing psychrophilic oil-degrading microbes and nutrients was sprayed twice daily over diesel-contaminated soil at 6 °C. Removal efficiencies in total petroleum hydrocarbon (TPH) at 30 days were 46.3% for landfarming and 73.7% for foam-spraying. The first-order kinetic biodegradation rates for landfarming and foam-spraying were calculated as 0.019 d(-1) and 0.044 d(-1), respectively. Foam acted as an insulating medium, keeping the soil 2 °C warmer than ambient air. Sprayed foam was slowly converted to aqueous solution within 10-12h and infiltrated the soil, providing microbes, nutrients, water, and air for bioaugmentation. Furthermore, surfactant present in the aqueous solution accelerated the dissolution of oil from the soil, resulting in readily biodegradable aqueous form. Significant reductions in hydrocarbon concentration were simultaneously observed in both semi-volatile and non-volatile fractions. As the initial soil TPH concentration increased, the TPH removal rate of the foam-spraying method also increased.

Enhanced isolation and culture of highly efficient psychrophilic oil-degrading bacteria from oil-contaminated soils in South Korea.

It is known that isolation of oil-degrading bacterial strains is difficult at low temperatures, and the biodegradation efficiency of oil-contaminated soil is significantly reduced in cold weather. In this study, 14 strains were isolated from oil-contaminated soil that grew well at 10°C by using a newly developed culture method. 11 of the 14 isolates were successfully cultured in mineral salts medium containing 1,500 ppm of oil components, 500 ppm each kerosene, gasoline, and diesel as carbon sources, at 10°C for 2 weeks. The oil degradation efficiencies of these 11 isolates ranged from 36% to 100%, as measured by total petroleum hydrocarbon (TPH) degradation analyses. Three strains (Pseudomonas simiae G1-10O, P. taiwanensis Y1-4, and P. koreensis Gwa2) displayed complete degradation (100%), and six others (R frederiksbergensis G2-2, P arsenicoxydans Y2-1, R umsongensis Gwa3, P. migulae Gwa5, RhodococcusjialingiaeY 1-l , and R. qingshengii Y2-2) showed relatively high degradation efficiencies (> 70%). This study suggests that these isolates can be effectively utilised in thetreatment of oil-contaminated soil in landfarming, especially during winter.

Comparative study of rhizobacterial community structure of plant species in oil-contaminated soil.

In this study, the identity and distribution of plants and the structure of their associated rhizobacterial communities were examined in an oil-contaminated site. The number of plant species that formed a community or were scattered was 24. The species living in soil highly contaminated with total petroleum hydrocarbon (TPH) (9,000-4,5000 mg/g-soil) were Cynodon dactylon, Persicaria lapathifolia, and Calystegia soldanella (a halophytic species). Among the 24 plant species, the following have been known to be effective for oil removal: C. dactylon, Digitaria sanguinalis, and Cyperus orthostachyus. Denaturing gradient gel electrophoresis (DGGE) profile analysis showed that the following pairs of plant species had highly similar (above 70%) rhizobacterial community structures: Artemisia princeps and Hemistepta lyrata; C. dactylon and P. lapathifolia; Carex kobomugi and Cardamine flexuosa; and Equisetum arvense and D. sanguinalis. The major groups of rhizobacteria were Betaproteobacteria, Gamma-proteobacteria, Chloroflexi, Actinobacteria, and unknown. Based on DGGE analysis, P. lapathifolia, found for the first time in this study growing in the presence of high TPH, may be a good species for phytoremediation of oil-contaminated soils and in particular, C. soldanella may be useful for soils with high TPH and salt concentrations. Overall, this study suggests that the plant roots, regardless of plant species, may have a similar influence on the bacterial community structure in oil-contaminated soil.

Effects of crude oil, oil components, and bioremediation on plant growth.

The phytotoxic effects of crude oil and oil components on the growth of red beans (Phaseolus nipponesis OWH1) and corn (Zea mays) was investigated. In addition, the beneficial effects of bioremediation with the oil-degrading microorganism, Nocardia sp. H17-1, on corn and red bean growth in oil-contaminated soil was also determined. It was found that crude oil-contaminated soil (10,000mg/kg) was phytotoxic to corn and red beans. In contrast, obvious phytotoxicity was not observed in soils contaminated with 0-1000 mg/kg of aliphatic hydrocarbons such as decane (C10) and eicosane (C20). Phytotoxicity was observed in soils contaminated with 10-1000mg/kg of the poly aromatic hydrocarbons (PAHs) naphthalene, phenanthrene, and pyrene. It was observed that phytotoxicity increased with the number of aromatic rings, and that corn was more sensitive than red beans to PAH-contaminated soil. Bioremediation with Nocardia sp. H17-1 reduced phytotoxicity more in corn than in red bean, suggesting that this microbial species might degrade PAHs to some degree.

Uptake and phytotoxicity of TNT in onion plant.

The uptake of 14C-2,4,6-trinitrotoluene (TNT) in hydroponics was studied using onion plants. Of the total TNT mass (5 microM concentration), 75% was in the roots, 4.4% in the leaves, and 21% in the external solution at 2 days. The percent distribution in roots was lower with higher concentration in the external solution, but in leaves it was comparable at all concentrations (5-500 microM). Root concentration factor (RCF) in hydroponics was more than 85 in constant hydroponic experiment (CHE) at 5 microM and 150 in nonconstant hydroponic experiment (NHE) at 5 microM. The maximum RCF values in the hydroponic system were greater with lower solution concentration. Transpiration stream concentration factor (TSCF) values in the present study (NHE only: 0.31-0.56) were relatively similar to the values with predicted values (0.43-0.78), increasing with higher external TNT concentration. All predicted values for RCF and TSCF were derived in the literature from equations using logKow (log10 octanol-water partition coefficient). For phytotoxicity tested in hydroponics and wet paper method, 500 microM was toxic to onion plant, 50 microM was nontoxic for plant growth but limited the transpiration rate, and 5 microM was nontoxic as control.