Phytases are a group of digestive enzymes which are commonly used as feed enzymes. These enzymes are used exogenously in the feeds of monogastric animals thereby it improves the digestibility of phosphorous and thus reduces the negative impact of inorganic P excretion on the environment. Even though these enzymes are widely distributed in many life forms, microorganisms are the most preferred and potential source of phytase. Despite the extensive availability of the phytase-producing microbial consortia, only a few microorganisms have been known to be exploited at industrial level. The high costs of the enzyme along with the incapability to survive high temperatures followed by the poor storage stability are noted to be the bottleneck in the commercialization of enzymes. For this reason, besides the conventional fermentation approaches, the applicability of cloning, expression studies and genetic engineering has been implemented for the past few years to accomplish the abovesaid benefits. The site-directed mutagenesis as well as knocking out have also validated their prominent role in microbe-based phytase production with enhanced levels. The present review provides detailed information on recent insights on the modification of phytases through heterologous expression and protein engineering to make thermostable and protease-resistant phytases.

Microplastics (MPs) represent a matter of growing concern for the marine environment. Their ingestion has been documented in several species worldwide, but the impact of specific anthropogenic activities remains largely unexplored. In this study, MPs were characterized in different benthic fish sampled after 2.5 years of huge engineering operations for the parbuckling project on the Costa Concordia wreck at Giglio Island. Fish collected in proximity of the wreck showed a high ingestion of microplastics compared to both fish from a control area and values reported worldwide. Also the elevated percentage of nylon, polypropylene lines and the presence of polystyrene are quite unusual for marine organisms sampled in natural field conditions, thus supporting the possible relationship of ingested microplastics with maritime operations during wreck removal. On the other hand, the use of transplanted mussels revealed a lower frequency of ingested MPs, and did not discriminate differences between the wreck and the control area. Some variations were observed in terms of typology and size of particles between surface- and bottom-caged mussels highlighting the influence of a different distribution of MPs along the water column. In conclusion, this study demonstrated that MPs pollution in the area of Costa Concordia was more evident on benthonic environment than on seawater column, providing novel insights on the possibility of using appropriate sentinel organisms for monitoring specific anthropogenic sources of MPs pollution in the marine environment.

The use of zero-valent iron nanoparticles (nZVI) has been advocated for the remediation of both soils and groundwater. A key parameter affecting nZVI remediation efficacy is the mobility of the particles as this influences the reaction zone where remediation can occur. However, by engineering nZVI particles with increased stability and mobility we may also inadvertently facilitate nZVI-mediated contaminant transport away from the zone of treatment. Previous nZVI mobility studies have often been limited to model systems as the presence of background Fe makes detection and tracking of nZVI in real systems difficult. We overcame this problem by synthesising Fe-59 radiolabelled nZVI. This enabled us to detect and quantify the leaching of nZVI-derived Fe-59 in intact soil cores, including a soil contaminated by Chromated-Copper-Arsenate. Mobility of a commercially available nZVI was also tested. The results showed limited mobility of both nanomaterials; <1% of the injected mass was eluted from the columns and most of the radiolabelled nZVI remained in the surface soil layers (the primary treatment zone in this contaminated soil). Nevertheless, the observed breakthrough of contaminants and nZVI occurred simultaneously, indicating that although the quantity transported was low in this case, nZVI does have the potential to co-transport contaminants. These results show that direct injection of nZVI into the surface layers of contaminated soils may be a viable remediation option for soils such as this one, in which the mobility of nZVI below the injection/remediation zone was very limited. This Fe-59 experimental approach can be further extended to test nZVI transport in a wider range of contaminated soil types and textures and using different application methods and rates. The resulting database could then be used to develop and validate modelling of nZVI-facilitated contaminant transport on an individual soil basis suitable for site specific risk assessment prior to nZVI remediation.

Understanding Pb removal from the translocation stream is vital to engineering Pb hyperaccumulation in above ground organs, which would enhance the economic feasibility of Pb phytoextraction technologies. We investigated Cu, Pb, Sb and Zn distributions in Hay-scented fern (Dennstaedtia punctilobula) rhizomes on shooting range soils by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS), analyzing digested rhizomes, stems, and fronds using ICP-MS. Nutrients Cu and Zn concentrated in fronds while toxic elements Pb and Sb did not, showing potential Pb and Sb sequestration in the rhizome. Frond and rhizome concentration of Pb was 0.17 ± 0.10% and 0.32 ± 0.21% of dry biomass, respectively. The ²⁰⁸Pb/¹³C and ¹²¹Sb/¹³C determined by LA-ICP-MS increased from inner sclerotic cortex to the epidermis, while Pb concentrated in the starchy cortex only in contaminated sites. These results suggest that concentration dependent bioaccumulation in the rhizome outer cortex removes Pb from the vascular transport stream.

Thousands of hectares of wetlands are created annually because wetlands provide beneficial ecosystem services. Wetlands are also key sites for production of the bioaccumulative neurotoxin methylmercury (MeHg), but little is known about MeHg production in created systems. Here, we studied methylmercury in sediment, water, and invertebrates in created wetlands of various ages. Sediment MeHg reached 8 ng g⁻¹ in the newest wetland, which was significantly greater than in natural, control wetlands. This trend was mirrored in several invertebrate taxa, whose concentrations reached as high as 1.6 μg g⁻¹ in the newest wetland, above levels thought to affect reproduction in birds. The MeHg concentrations in created wetland invertebrate taxa generally decreased with increasing wetland age, possibly due to a combination of deeper anoxia and less organic matter accumulation in younger wetlands. A short-term management intervention and/or improved engineering design may be necessary to reduce the mercury-associated risk in newly created wetlands.

Heavy metal pollution is a serious environmental problem globally, particularly in mines and tailings ponds. In this study, based on laboratory and field tests, the migration of heavy metal contaminants in a tailings pond and the retention behavior of a compacted bentonite engineered barrier system on the heavy metal contaminants were analyzed by a numerical simulation. The results demonstrate that the hydraulic conductivity of compacted bentonite is lower than that of the tailings from the laboratory tests. The hydraulic conductivity of the tailings sand decreased with an increase in the dry density and increased with an increase in the concentration of the chemical solution, which could be attributed to the large amounts of fine-grained soil contained in the tailings, according to the grain size distribution test. The hydraulic conductivity of the tailings from the engineering geological survey was between 2.0 × 10−6 and 9.0 × 10−5 m/s, and followed the order: tail coarse sand ＞ tail silty sand ＞ tail medium sand ＞ tail fine silt. The numerical simulation of the seepage could satisfactorily describe the actual working condition of the tailings dam. With the groundwater seepage, the migration range of the heavy metal contaminant in the researched tailings pond reached a maximum of 45 m for 5 years. The retention efficiencies of the 0.2 m engineered barrier against the heavy metal contaminant for 15 and 30 years were 45.4% and 57.2%, respectively. Moreover, the retention efficiency would exceed 87% when the engineered barrier thickness is increased to 0.5 m. The results of model validation show that the calculated results are in good agreement with the measured ones. These findings can provide effective ideas for the prevention and control of environmental pollution in mines and tailings ponds.

In this study, a proposed integrated high-gravity technology for air pollution control, CO2 capture, and alkaline waste utilization was comprehensively evaluated from engineering, environmental, and economic perspectives. After high-gravity technology and coal fly ash (CFA) leaching processes were integrated, flue gas air emissions removal (e.g., sulfate dioxide (SO2), nitrogen oxides (NOx), total suspended particulates (TSP)) and CO2 capture were studied. The CFA, which contains calcium oxide and thus, had high alkalinity, was used as an absorbent in removing air pollution residues. To elucidate the availability of technology for pilot-scale high-gravity processes, the engineering performance, environmental impact, and economic cost were simultaneously investigated. The results indicated that the maximal CO2, SO2, NOx, and TSP removal efficiencies of 96.3 ± 2.1%, 99.4 ± 0.3%, 95.9 ± 2.1%, and 83.4 ± 2.6% were respectively achieved. Moreover, a 112 kWh/t-CO2 energy consumption for a high-gravity process was evaluated, with capture capacities of 510 kg CO2 and 0.468 kg NOx per day. In addition, the fresh, water-treated, acid-treated, and carbonated CFA was utilized as supplementary cementitious materials in the blended cement mortar. The workability, durability, and compressive strength of 5% carbonated CFA blended into cement mortar showed superior performance, i.e., 53 MPa ±2.5 MPa at 56 days. Furthermore, a higher engineering performance with a lower environmental impact and lower economic cost could potentially be evaluated to determine the best available operating condition of the high-gravity process for air pollution reduction, CO2 capture, and waste utilization.

In this report, we establish proof-of-principle demonstrating for the first time genetic engineering of a photoautotrophic microorganism for bioremediation of naturally occurring cyanotoxins. In model cyanobacterium Synechocystis sp. PCC 6803 we have heterologously expressed Sphingopyxis sp. USTB-05 microcystinase (MlrA) bearing a 23 amino acid N-terminus secretion peptide from native Synechocystis sp. PCC 6803 PilA (sll1694). The resultant whole cell biocatalyst displayed about 3 times higher activity against microcystin-LR compared to a native MlrA host (Sphingomonas sp. ACM 3962), normalized for optical density. In addition, MlrA activity was found to be almost entirely located in the cyanobacterial cytosolic fraction, despite the presence of the secretion tag, with crude cellular extracts showing MlrA activity comparable to extracts from MlrA expressing E. coli. Furthermore, despite approximately 9.4-fold higher initial MlrA activity of a whole cell E. coli biocatalyst, utilization of a photoautotrophic chassis resulted in prolonged stability of MlrA activity when cultured under semi-natural conditions (using lake water), with the heterologous MlrA biocatalytic activity of the E. coli culture disappearing after 4 days, while the cyanobacterial host displayed activity (3% of initial activity) after 9 days. In addition, the cyanobacterial cell density was maintained over the duration of this experiment while the cell density of the E. coli culture rapidly declined. Lastly, failure to establish a stable cyanobacterial isolate expressing native MlrA (without the N-terminus tag) via the strong cpcB560 promoter draws attention to the use of peptide tags to positively modulate expression of potentially toxic proteins.

Disinfection byproducts (DBPs) generated by ballast water treatment have become a concern worldwide because of their potential threat to the marine environment. Predicting the relative DBP concentrations after disinfection could enable better control of DBP formation. However, there is no appropriate method of evaluating DBP formation in a full-scale ballast water treatment system (BWTS). In this study, multiple regression models were developed for predicting the dibromochloromethane (DBCM) and bromoform (TBM) concentrations produced by an emergency BWTS using field experimental data from ballast water treatments conducted at Dalian Port, China. Six combinations of independent variables [including several water parameters and/or the total residual oxidant (TRO) concentration] were evaluated to construct mathematical prediction formulas based on a polynomial linear model and logarithmic regression model. Further, statistical analyses were performed to verify and determine the appropriate mathematical models for DBCM and TBM formation, which were ultimately validated using additional field experimental data. The polynomial linear model with four variables (temperature, salinity, chlorophyll, and TRO) and the logarithmic regression model with seven variables (temperature, salinity, dissolved oxygen, pH, turbidity, chlorophyll, and TRO) exhibited good reproducibility and could be used to predict the DBCM and TBM concentrations, respectively. The validation results indicated that the developed models could accurately predict DBP concentrations, with no significant statistical difference from the measured values. The results of this work could provide a theoretical basis and data reference for ballast water treatment control in engineering applications of emergency BWTSs.

Introducing of earthworms to constructed wetlands (CWs) has been considered as a new approach to solve the clogging problems in the long-established systems. Despite its potential advantage, the correlational researches are still in the stage of preliminary observation and speculation. This paper presents a comprehensive and in-depth research about the positive effects of earthworms (Eisenia foetida) on clog matter (CM) reduction through different pathways, including in vivo metabolism and uptake, conversion, transport, and promotion of microorganism quantities. The results showed that the metabolism and uptake by Eisenia foetida could effectively reduce the CM content at an average removal rate of 0.155 mg g−1 d−1, which was obviously higher than the rate of CM decomposition by microorganisms alone. Through the metabolism of earthworms, the amounts of proteins and polysaccharides in CM were decreased, while the amounts of humin and nucleic acids were increased. Simultaneously, the viscosity of CM was reduced by 0.0082 mPa s g−1 d−1, and the quantity of microorganisms was increased by 0.0109 mg g−1 d−1, which finally made the treated CM can be easily washed away and decomposed. Furthermore, earthworms could reduce the CM content in the clogging layer by transporting the metabolic products out. A regression model was further performed for describing the interaction between earthworm and CM. The simulated value of porosity fitted well with the measured one, suggesting that the earthworms can increase the substrate porosity at a rate of 0.33 mL g−1 d−1. This study quantitively depicted the mechanisms of earthworms on the decrement of CM content in CWs, which is of great benefit for the engineering management of constructed wetlands in the future. We also proposed that the density of introduced earthworms should exceed a certain threshold for effectively increasing the substrate porosity and solving the clogging problems.