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Antibiotic resistance genes (ARGs) in soil pose a major challenge to global environment and health. The development of effective technologies to reduce their negative effects has implications for maintaining soil health and human health. Biochar would be a suitable control material due to its characteristics of high carbon content, large surface area, excellent adsorption capacity, and economic advantages. There are three mechanisms underlying its negative effects on the abundance of ARGs: 1) adsorption of certain pollutants (e.g., antibiotics and heavy metals) to reduce the co-selective pressure of ARGs; 2) alteration of microbial composition through altering soil physico-chemical properties, and thereby limiting the ability of bacteria to undergo horizontal transfer of ARGs; 3) direct impairment of horizontal gene transfer by the adsorption of horizontal transfer vectors such as plasmids, transposons, and integrons. However, the negative effect of biochar depends on the source of material, pyrolysis process, and its amount added. Furthermore, field aging of biochar may reduce its ability to block ARGs. Endogenous contaminants of biochar, such as polycyclic aromatic hydrocarbons and heavy metals, may cause the enrichment of specific antibiotic-resistant bacteria in the environment or induce horizontal gene transfer. In further studies, suitable biochar should be selected according to soil environments, and biochar aging control measures should be taken to improve its retarding effect on ARGs.

Freezing method assists calcium hypochlorite for synergistically promoting methane production from sludge anaerobic digestion.

Anaerobic digestion is widely considered to be a promising technology for waste activated sludge (WAS) treatment, by which sludge stabilization and resource recovery are simultaneously achieved. The poor reaction efficiency however hinders the large-scale applications of WAS anaerobic digestion technology. This study reported an efficient sludge pretreatment method by combining freezing with calcium hypochlorite (Ca(ClO)2) for enhancing the anaerobic digestion efficiency. Experimental data showed that the optimal combination was freezing at -20 °C coupled with 0.075 g/g VSS (volatile suspended solids) Ca(ClO)2, by which the maximum biomethane production of 274.4 ± 8.2 mL/g VSS was realized, 1.62 times higher than that of the control. Model-based analysis demonstrated that higher potential and rate for methane production were attained by the combined pretreatment. Mechanism analysis revealed that the extracellular polymeric substances (EPS) and microbial cells were both effectively destructed when treated by combined freezing and Ca(ClO)2, and more dissolved organics were generated in consequence. Microbial analysis demonstrated that the co-treated reactor enriched more functional microbes (such as Methanosaeta, Methanosarcina and Candidatus_Methanofastidiosum) responsible for biomethane generation than that of the control. Furthermore, the number of fecal coliform was largely reduced in co-treated reactor. As the correlation between sludge anaerobic digestion performance and numerous pretreatment parameters was systematically revealed, this study can provide important references for engineers when applying the combined freezing and Ca(ClO)2 technology in practical engineering.

Effects of enriched planting of native tree species on surface water flow, sediment, and nutrient losses in a Eucalyptus plantation forest in southern China.

Enriched planting of native tree species in monoculture plantation forests is a commonly recommended forest practice. However, its effect on various ecological processes is generally lacking. Here, we carried out an experiment in a 16-year-old Eucalyptus plantation in South China to assess the effects of enriched planting of native tree species on surface water, soil erosion and nutrient losses. Two treatments were conducted in 2008: (1) enriched planting of native broadleaved tree species with uniform thinning of 60% of Eucalyptus trees (TEP); and (2) enriched planting of native broadleaved tree species without thinning (NEP). The original Eucalyptus plantation stands was used as the control (CK). Runoff plots (total n=9, 3 for each treatment or CK) were established in 2009, and surface water flow, sediment, nitrogen (N) and phosphorus (P) losses were monitored from major rainfall events in 2010-2012. Results showed that enriched planting in Eucalyptus plantation significantly reduced surface water flow and soil erosion. Compared with CK, TEP and NEP reduced annual surface water flow by 29-43% and 11-16%, and reduced annual soil erosion by 38-54% and 20-33% throughout the study period, respectively. TEP and NEP had significantly lower annual mean concentrations of N and P in surface water. Compared with CK, TEP reduced annual N and P losses through surface water by 42-60% and 44-64%, respectively, while NEP reduced them by 25-28% and 24-34%, respectively. N and P losses were significantly related to surface water flow. Between the two treatments, TEP was better for retaining water and soil, and for preventing nutrient loss. These results clearly demonstrated that the enriched planting of native tree species effectively retained surface water and nutrients.

Integrated application of sewage sludge, earthworms and Jatropha curcas on abandoned rare-earth mine land soil.

Improving soil fertility is a critical component of abandoned rare-earth mine land (ARL) revegetation. To study the effects of sewage sludge (SS), earthworms, and Jatropha curcas in ARL revegetation, SS (40% in mass ratio) and earthworms (0, 40, 60, and 80 individual adult Eisenia fetida kg-1) were applied to abandoned rare-earth mine land soil (ARLS) and then J. curcas was grown in a potting experiment. The organic carbon, nutrients (N, P, K) and heavy metals (HMs; Cd, Cu, Zn) contents in ARLS and the biomass and nutrients uptake in J. curcas were significantly increased by SS amendment. Application of 80 individual E. fetida kg-1 significantly increased availability of P and K in SS-amended ARLS relative to other treatments. Earthworms increased the height, ground diameter and biomass of J. curcas, but the promotion of biomass became weaker as earthworm density increased. After J. curcas harvest, the contents of Cd, Cu and Zn in SS-amended ARLS were reduced by 15%, 23%, and 19%, respectively. With the joint application of J. curcas and earthworms, a much larger decrease in contents of Cd (34-40%), Cu (31-44%) and Zn (24-29%) in SS-amended ARLS were observed, and the HMs ecological risks were reduced from "moderate potential" to "low potential". Moreover, J. curcas and earthworms together exerted more reduction in the exchangeable fraction HMs in SS-amended ARLS than J. curcas alone. Our results suggest that the integrated application of SS, earthworms and J. curcas is an effective approach for ARL revegetation.

Effects of landscape plant species and concentration of sewage sludge compost on plant growth, nutrient uptake, and heavy metal removal.

Landscape plants have great potentials in heavy metals (HMs) removal as sewage sludge compost (SSC) is increasingly used in urban forestry. We hypothesize that woody plants might perform better in HMs phytoremediation because they have greater biomass and deeper roots than herbaceous plants. We tested the differences in growth responses and HMs phytoremediation among several herbaceous and woody species growing under different SSC concentrations through pot experiments. The mixing percentage of SSC with soil at 0%, 15%, 30%, 60, and 100% were used as growth substrate for three woody (Ficus altissima Bl., Neolamarckia cadamba (Roxb.) Bosser, and Bischofia javanica Bl.) and two herbaceous (Alocasia macrorrhiza (L.) G. Don and Dianella ensifolia (L.) DC) plants. Results showed that the biomass, relative growth rate, and nutrient uptake for all plants increased significantly at each SSC concentration compared to the control; woody plants had higher biomass and nutrient use efficiency than herbaceous plants. All plants growing in SSC-amended soils accumulated appreciable amounts of HMs and reduced the contents of HMs present in the substrates. The woody plants were generally more effective than herbaceous plants in potentials of HMs phytoextraction, but A. macrorrhiza showed higher bioconcentration and translocation of Cu and Zn and D. ensifolia had higher bioconcentration and translocation of Cd than woody plants. The optimal application concentrations were 30% or less for woody plants and 15% for herbaceous plants for plant growth and ecological risk control, respectively. Intercropping suitable woody and herbaceous landscape plants in urban forestry might have promising potentials to minimize the ecological risks in the phytoremediation of SSC.

Alleviation of heavy metal phytotoxicity in sewage sludge by vermicomposting with additive urban plant litter.

The handling of sewage sludge (SS) and urban plant litter (UPL) has become an important concern. Immobilizing heavy metals (HMs) is regarded as a necessary process for recycling SS in agriculture and forestry. Here, HM removal and HM phytotoxicity in SS during vermicomposting with different additive UPLs was investigated. The results show that vermicomposting with additive UPL significantly reduced the content of HMs, and increased organic carbon content and the proportion of macroaggregates in SS. This process also significantly immobilized HMs by mainly transforming extractable and reducible HMs into residual products. The litters of Dracontomelon duperreanum and Bauhinia purpurea increased oxidizable HMs in SS and the accumulation capacity of HMs of earthworms during vermicomposting. The Cd content in vermicomposts with the B. purpurea litter addition was decreased by 31% relative to the initial SS. Maize in vermicomposts with UPL additions, especially with B. purpurea litter, exhibited significan5tly higher seed germination rates, seedling biomass, root activity, and a lower accumulation of HMs than in SS compost without UPL additions. These results suggest that vermicomposting with additive UPL can alleviate the phytotoxicity of HMs in SS and provides a new method for simultaneously recycling SS and UPL.

Municipal sewage sludge compost promotes Mangifera persiciforma tree growth with no risk of heavy metal contamination of soil.

Application of sewage sludge compost (SSC) as a fertilizer on landscaping provides a potential way for the effective disposal of sludge. However, the response of landscape trees to SSC application and the impacts of heavy metals from SSC on soil are poorly understood. We conducted a pot experiment to investigate the effects of SSC addition on Mangifera persiciforma growth and quantified its uptake of heavy metals from SSC by setting five treatments with mass ratios of SSC to lateritic soil as 0%:100% (CK), 15%:85% (S15), 30%:70% (S30), 60%:40% (S60), and 100%:0% (S100). As expected, the fertility and heavy metal concentrations (Cu, Zn, Pb and Cd) in substrate significantly increased with SSC addition. The best performance in terms of plant height, ground diameter, biomass and N, P, K uptake were found in S30, implying a reasonable amount of SSC could benefit the growth of M. persiciforma. The concentrations of Cu, Pb and Cd in S30 were insignificantly different from CK after harvest, indicating that M. persiciforma reduced the risk of heavy metal contamination of soil arising from SSC application. This study suggests that a reasonable rate of SSC addition can enhance M. persiciforma growth without causing the contamination of landscaping soil by heavy metals.

Screening and structural and functional investigation of a novel ferritin from Phascolosoma esculenta.

Ferritins are primary iron storage proteins and play a crucial role in iron storage and detoxification. Yeast two-hybrid method was employed to screen the cDNA library of Phascolosoma esculenta. Sequence of positive colony FER147 was analyzed. The higher similarity and conserved motifs for ferritin indicated that it belonged to a new member of ferritin family. The interaction between Ferritin and Fer147 was further confirmed through co-immunoprecipitation. The pET-28a-FER147 prokaryotic expression vector was constructed. The expressed recombinant Fer147 was then isolated, purified, and refolded. When ferritins were treated by different heavy metals, several detection methods, including scanning electron microscopy (SEM), circular dichroism (CD), and inductively coupled plasma-mass spectrometry (ICP-MS) were applied to examine the structures and functions of the new protein Fer147, recombinant P. esculenta ferritin (Rferritin), and natural horse-spleen ferritin (Hferritin). SEM revealed that the three ferritin aggregates changed obviously after different heavy metals treatment, meanwhile, a little different in aggregates were detected when the ferritins were trapped by the same heavy metal. Hence, changes in aggregation structure of the three proteins are related to the nature of the different heavy metals and the interaction between the heavy metals and the three ferritins. CD data suggested that the secondary structure of the three ferritins hardly changed after different heavy metals were trapped. ICP-MS revealed that the ferritins exhibit different enrichment capacities for various heavy metals. In particular, the enrichment capacity of the recombinant Fer147 and Rferritin is much higher than that of hferritin.

Preparation and representation of recombinant Mn-ferritin flower-like spherical aggregates from marine invertebrates.

Ferritin has important functions in the transition and storage of toxic metal ions, but its regulation and function in many invertebrate species are still largely unknown. In our previous work, the cDNA sequence of Sinonovacula constricta, Apostichopus japonicas and Acaudina leucoprocta were constructed and efficiently expressed in E. Coli BL21 under IPTG induction. In this follow-up study, the recombinant ferritins were exposed to heavy metal manganese. The manganese concentration levels in three recombinant ferritins were greater than horse spleen ferritin (HSF). Compared with HSF, the amount of manganese enrichment in the three recombinant ferritins was 1.75-fold, 3.25-fold and 2.42-fold increases in ScFER, AjFER, and AlFER, respectively. After phosphate stimulation, the concentration of manganese increased and was higher than the ordinary dialysis control groups. The ScFER was four times its baseline value. The AjFER and AlFER were 1.4- and 8-fold higher, respectively. The AlFER sample stimulated by phosphate was 22-fold that of HSF. The morphologies of the resulting Mn-Ferritin from different marine invertebrates were characterized with scanning electron microscopy. Surface morphologies were lamella flower-like and are consistent with changes in surface morphologies of the standard Mn-HSF. Invertebrate recombinant ferritin and HSF both can uptake manganese. We found that the structure of A. leucoproctarecombinant Mn-Ferritin aggregate changed over time. The surface formed lamella flower-like aggregate, but gradually merged to create a relatively uniform plate-like phase of aggregate spherically and fused without clear boundaries.