Temperature-phased anaerobic sludge digestion effectively removes antibiotic resistance genes in a full-scale wastewater treatment plant.

Sludge is a major by-product and the final reservoir of antibiotic resistance genes (ARGs) in wastewater treatment plants (WWTPs). Temperature-phased anaerobic digestion (TPAD), consisting of thermophilic anaerobic digestion (AD) (55 °C) and mesophilic AD processes (37 °C), has been implemented in WWTPs for sludge reduction while improving the biomethane production. However, the impact of TPAD on the ARGs' fate is still undiscovered in lab-scale experiments and full-scale WWTPs. This study, for the first time, investigated the fate of ARGs during the TPAD process across three seasons in a full-size WWTP. Ten typical ARGs and one integrase gene of class 1 integron (intI1) involving ARGs horizontal gene transfer were examined in sludge before and after each step of the TPAD process. TPAD reduced aac(6')-Ib-cr, blaTEM, drfA1, sul1, sul2, ermb, mefA, tetA, tetB and tetX by 87.3-100.0 %. TPAD reduced the overall average absolute abundance of targeted ARGs and intI1 by 92.39 % and 92.50 %, respectively. The abundance of targeted ARGs in sludge was higher in winter than in summer and autumn before and after TPAD. During the TPAD processes, thermophilic AD played a major role in the removal of ARGs, contributing to >60 % removal of ARGs, while the subsequent mesophilic AD contributed to a further 31 % removal of ARGs. The microbial community analysis revealed that thermophilic AD reduced the absolute abundance of ARGs hosts, antibiotic resistant bacteria. In addition, thermophilic AD reduced the abundance of the intI1, while the intI1 did not reproduce during the mesophilic AD, also contributing to a decline in the absolute abundance of ARGs in TPAD. This study demonstrates that TPAD can effectively reduce the abundance of ARGs in sludge, which will suppress the transmission of ARGs from sludge into the natural environment and deliver environmental and health benefits to our society.

Occurrence, fate, and remediation for per-and polyfluoroalkyl substances (PFAS) in sewage sludge: A comprehensive review.

Addressing per-and polyfluoroalkyl substances (PFAS) contamination is an urgent environmental concern. While most research has focused on PFAS contamination in water matrices, comparatively little attention has been given to sludge, a significant by-product of wastewater treatment. This critical review presents the latest information on emission sources, global distribution, international regulations, analytical methods, and remediation technologies for PFAS in sludge and biosolids from wastewater treatment plants. PFAS concentrations in sludge matrices are typically in hundreds of ng/g dry weight (dw) in developed countries but are rarely reported in developing and least-developed countries due to the limited analytical capability. In comparison to water samples, efficient extraction and cleaning procedures are crucial for PFAS detection in sludge samples. While regulations on PFAS have mainly focused on soil due to biosolids reuse, only two countries have set limits on PFAS in sludge or biosolids with a maximum of 100 ng/g dw for major PFAS. Biological technologies using microbes and enzymes present in sludge are considered as having high potential for PFAS remediation, as they are eco-friendly, low-cost, and promising. By contrast, physical/chemical methods are either energy-intensive or linked to further challenges with PFAS contamination and disposal. The findings of this review deepen our comprehension of PFAS in sludge and have guided future research recommendations.

Enhancing methane production from anaerobic digestion of secondary sludge through lignosulfonate addition: Feasibility, mechanisms, and implications.

This study explores the feasibility of using lignosulfonate, a byproduct of the pulp and paper industry, to facilitate sludge anaerobic digestion. Biochemical methane potential assays revealed that the maximum methane production was achieved at 60 mg/g volatile solids (VS) lignosulfonate, 22.18 % higher than the control. One substrate model demonstrated that 60 mg/g VS lignosulfonate boosted the hydrolysis rate, biochemical methane potential, and degradation extent of secondary sludge by 19.12 %, 21.87 %, and 21.11 %, respectively, compared to the control. Mechanisms unveiled that lignosulfonate destroyed sludge stability, promoted organic matter release, and enhanced subsequent hydrolysis, acidification, and methanogenesis by up to 31.30 %, 74.42 % and 28.16 %, respectively. Phytotoxicity assays confirmed that lignosulfonate promoted seed germination and root development of lettuce and Chinese cabbage, with seed germination index reaching 170 ± 10 % and 220 ± 22 %, respectively. The findings suggest that lignosulfonate addition offers a sustainable approach to sludge treatment, guiding effective management practices.

Climate change impacts on wastewater infrastructure: A systematic review and typological adaptation strategy.

Wastewater infrastructures play an indispensable role in society's functioning, human production activities, and sanitation safety. However, climate change has posed a serious threat to wastewater infrastructures. To date, a comprehensive summary with rigorous evidence evaluation for the impact of climate change on wastewater infrastructure is lacking. We conducted a systematic review for scientific literature, grey literature, and news. In total, 61,649 documents were retrieved, and 96 of them were deemed relevant and subjected to detailed analysis. We developed a typological adaptation strategy for city-level decision-making for cities in all-income contexts to cope with climate change for wastewater structures. 84% and 60% of present studies focused on the higher-income countries and sewer systems, respectively. Overflow, breakage, and corrosion were the primary challenge for sewer systems, while inundation and fluctuation of treatment performance were the major issues for wastewater treatment plants. In order to adapt to the climate change impact, typological adaptation strategy was developed to provide a simple guideline to rapidly select the adaptation measures for vulnerable wastewater facilities for cities with various income levels. Future studies are encouraged to focus more on the model-related improvement/prediction, the impact of climate change on other wastewater facilities besides sewers, and countries with low or lower-middle incomes. This review provided insight to comprehensively understand the climate change impact on wastewater facilities and facilitate the policymaking in coping with climate change.

An auxin-like supermolecule to simultaneously enhance growth and cumulative eicosapentaenoic acid production in Phaeodactylum tricornutum.

Phaeodactylum tricornutum, a model alga, is well known for its ability to accumulate intracellular omega-3 eicosapentaenoic acid (EPA). However, P.tricornutum cells need to have a higher EPA content if they are to be used for industrial applications. In this study, an auxin-like supermolecule (SM) was synthesised and used for the cultivation of P. tricornutum. Results show that the addition of 1 ppm of SM significantly increased the P. tricornutum cell density and boosted the P. tricornutum biomass. The experimental group treated with 5 ppm of SM, had an EPA content of 31.7%, which was a 2.09-fold increase over the EPA content in the untreated group. Overall, our results demonstrated that SM can significantly improve the microalgal growth and EPA accumulation in P. tricornutum, providing a feasible strategy to achieve efficient and cost-effective EPA production.

Enhancing the recombinant protein productivity of Yarrowia lipolytica using insitu fibrous bed bioreactor.

In this study, the ability of Yarrowia lipolytica to produce the recombinant lipase CalB from Candida antarctica, used as a model protein has been compared across different bioreactor processes using glycerol, a byproduct from the biodiesel industry as the main carbon source. Batch, pulsed fed-batch (PFB), and continuous fed-batch (CFB) strategies were first compared using classical stirred tank (STR) bioreactors in terms of biomass production, carbon source uptake, and lipase production. Additionally, an in situ fibrous bed bioreactor (isFBB) was developed using sugarcane bagasse as a cell immobilization support. The maximum lipase titer achieved using the isFBB culture mode was 38%, 33%, and 49% higher than those obtained using the batch, PFB, and CFB cultures, respectively. The lipase productivity in isFBB mode (142U/mL/h) was 1.4-fold higher than that obtained using batch free cell cultures. These results highlight that isFBB is an efficient technology for the production of recombinant enzymes.

Synergistic effect of alternating current and sulfate-reducing bacteria on corrosion behavior of X80 steel in coastal saline soil.

With the development of electrified railways and high-voltage transmission lines, it is often inevitable that buried metal structures are subjected to interference from the alternating current (AC) induced by the neighboring power facilities. Commonly found in soil, sulfate-reducing bacteria (SRB) have the capability to accelerate metal corrosion. In this paper, with electrochemical methods, surface analysis techniques, and weight-loss test, the influence of AC and SRB on the X80 steel corrosion behavior was explored in coastal saline soil. The results revealed that the 100 A m-2 AC inhibited the growth of the sessile and planktonic SRB cell. Under the action of 100 A m-2 AC, the metabolic activity of viable bacteria was enhanced, and the process of extracellular electron transfer was accelerated. When both AC and SRB were introduced, the maximum pit depth (76.2 µm) increased significantly to be 15 times higher than in the control condition (4.9 µm). Both SRB and AC played a role in enhancing corrosion. The corrosion rate of the AC-influenced specimen was far higher than that of the SRB-influenced specimen, while SRB and AC produced a synergistic effect on the enhanced corrosion of the specimen.

Effect of 'hand and foot acupuncture with twelve needles' on hemiplegia patients with 'qi deficiency and blood stasis' syndrome in the convalescent stage of Ischaemic stroke: study protocol for a randomised controlled trial.

BACKGROUND: Hemiplegia is a common sequela after stroke, and acupuncture is one of the most common physical therapies used to treat hemiplegia during the recovery stage after ischaemic stroke. 'Hand and foot acupuncture with twelve needles' is an acupuncture treatment performed after stroke. The principal objective of this study is to assess the efficacy and safety of 'hand and foot acupuncture with twelve needles' for hemiplegia in the convalescent stage of ischaemic stroke. METHODS: This is the protocol for a randomised, controlled clinical trial with two groups: a 'hand and foot acupuncture with twelve needles' group and a routine acupuncture group. A total of 208 participants will be randomly assigned to two different groups in a 1:1 ratio and will undergo conventional rehabilitation. Limb function will be evaluated by the simplified Fugl-Meyer assessment scale, Barthel Index, modified Ashworth scale and National Institute of Health stroke scale. The participants will be evaluated at baseline (on the day of enrolment) and followed up at 2 weeks, 1 month, 2 months and 3 months after enrolment. DISCUSSION: The results of this study will provide evidence on the effectiveness of 'hand and foot acupuncture with twelve needles' in the treatment of limb dysfunction that can be used for future evaluations. TRIAL REGISTRATION: Chictr.org.cn ChiCTR1900021774 . Registered on 8 March 2019.

Remaining Li-Content Dependent Structural Evolution during High Temperature Re-Heat Treatment of Quantitatively Delithiated Li-Rich Cathode Materials with Surface Defect-Spinel Phase.

Pre-extracting Li+ from Li-rich layered oxides by chemical method is considered to be a targeted strategy for improving this class of cathode material. Understanding the structural evolution of the delithiated material is very important because this is directly related to the preparation of electrochemical performance enhanced Li-rich material. Herein, we perform a high temperature reheat treatment on the quantitatively delithiated Li-rich materials with different amounts of surface defect-spinel phase and carefully investigate the structural evolution of these delithiated materials. It is found that the high temperature reheat treatment could cause the decomposition of the unstable surface defect-spinel structure, followed by the rearrangement of transition metal ions to form the thermodynamically stable phases, More importantly, we find that this process has high correlation with the remaining Li-content in the delithiated material. When the amount of extracted Li+ is relatively small (corresponding to the higher remaining Li-content), the surface defect-spinel phase could be dominantly decomposed into the LiMO2 (M = Ni, Co, and Mn) layered phase along with the significant improvement of electrochemical performance, and continuing to decrease remaining Li-content could lead to the emergence of M3O4-type spinel impurity embedding in the final product. However, when the extracted Li+ further achieves a certain amount, after the high temperature heat-treatment the Mn-rich Li2MnO3 phase (C2/m) could be separated from Ni-rich phases (including R3m, Fd3m, and Fm3m), thus resulting in a sharp deterioration of initial capacity and voltage. These findings suggest that reheating the delithiated Li-rich material to high temperature may be a simple and effective way to improve the predelithiation modification method, but first the amount of extracted Li+ should be carefully optimized during the delithiation process.

The growth and lutein accumulation in heterotrophic Chlorella protothecoides provoked by waste Monascus fermentation broth feeding.

Although the potential of heterotrophic microalgae served as a sustainable source for lutein, it was still crucial to formulate a suitable medium to offset the cost involved in algal biomass cultivation while improve inherent lutein productivity. The objective of this study was to investigate the feasibilities of waste Monascus fermentation broth medium (MFBM) toward heterotrophic Chlorella protothecoides-enriched lutein. The results indicated that C. protothecoides subjected to MFBM batch feeding achieved 7.1 g/L biomass and 7.27 mg/g lutein. The resulting lutein productivity (7.34 mg/L/day) represented 1.54-fold more than that of frequently used Basal medium. Concurrently, the effective metabolism and absorption of carbon, nitrogen, and phosphorus in MFBM by C. subellipsoidea cultivation make it easily complied with the permissible dischargeable limits for fermentation broth. When response to fed-batch culture mode, the biomass and lutein productivity peaked 20.4 g/L and 9.11 mg/L/day with concentrated MFBM feeding. Transcriptomics data hinted that MFBM feeding manipulated lutein biosynthesis key checkpoints (e.g., lycopene ß-cyclase and lycopene ε-cyclase) while accelerated energy pathways (e.g., glycolysis and TCA cycle) to contribute such high lutein productivity in C. protothecoides. These encouraging findings not only provided indications in applying nutrient-rich fermentation broth for affordable microalgae cultivation but also presented possibilities in linking algal high value-added products like lutein with high-efficient biological nutrition removal from industrial fermentation processing.

The potential growth and lipid accumulation in Coccomyxa subellipsoidea triggered by glucose combining with sodium acetate.

Carbon sources whether types or magnitudes were fateful in terms of stimulating growth and lipids accumulation in microalgae applied for biodiesel production. The set scenario of this work was to investigate the feasibilities of glucose (G) combining with sodium acetate (SA) carbon sources in enhancing biomass and lipid accumulation in Coccomyxa subellipsoidea. The results demonstrated that C. subellipsoidea subjected to the combination feeding of G (20 g/L) and SA (12 g/L) achieved the favorable biomass (5.22 g/L) and lipid content (52.16%). The resulting lipid productivity (388.96 mg/L/day) was 1.33- to 7.60-fold more than those of sole G or SA as well as other combinations of G and SA. Even though the total fatty acids of C. subellipsoidea cells treated with the optimal combination of G and SA showed no noticeable increment in comparison with sole G or SA, the proportion of monounsaturated C18:1 (over 48.69%) and the content of C18:3 (< 12%) were commendable in high-quality algal biodiesel production. Further, such fascinating lipid accumulation in C. subellipsoidea cells treated with G combining with SA might be attributed to that G promoted glycolysis as well as SA activated glyoxylate shunt and TCA cycle to synergistically provide sufficient acetyl-CoA precursors for lipid accumulation. These findings hinted the potential of the combination of carbon sources in enhancing the overall lipid productivity to offset alga-based biodiesel production cost and would guide other alga strains cultivation.

The enhanced biomass and lipid accumulation in Coccomyxa subellipsoidea with an integrated treatment strategy initiated by brewery effluent and phytohormones.

Brewery effluent (BE) as an appreciable and sustainable resource presented new possibilities in low-cost algal biomass production, whereas the relatively low essential macronutrients hindered extensive applications as growth medium for microalgae cultivation. The objective of this study was to investigate the feasibility of an integrated treatment strategy initiated by BE coupling phytohormones in augmenting biomass and lipid accumulation in Coccomyxa subellipsoidea. Results revealed that BE coupling synthetic 1-naphthaleneacetic acid (NAA) accomplished the favorable lipid productivity of 481.76 mg/L/days, representing 6.80- to 9.71-fold more than that of single BE as well as standard Basal media. BE coupling NAA feeding also heightened the proportions of C16-C18 fatty acids (over 96%) and mono-unsaturated C18:1 (approximate 45%) which were prone to high-quality biofuels-making. Such profound lipids accumulation might be attributable to that BE coupling NAA treatment drove most of metabolic flux (i.e. acetyl-CoA) derived from TCA cycle and glycolysis flowing into lipid accumulation pathway. Concurrently, the complete removal of total nitrogen and total phosphorus by C. subellipsoidea with assistance of NAA were easily complied with the permissible dischargeable limits for BE. These present results strongly demonstrated that BE coupling NAA was a potential feeding strategy in boosting algal lipid productivity and further provided great possibilities in linking affordable algal biomass production with high-efficient biological contaminants removal.

The augmented lipid productivity in an emerging oleaginous model alga Coccomyxa subellipsoidea by nitrogen manipulation strategy.

The lipid productivity controlled by both of biomass and lipid content was really crucial for economic-feasibility of microalgae-based biofuels production. This study attempted at augmenting lipid productivity in an emerging oleaginous model alga Coccomyxa subellipsoidea by different nitrogen manipulation including one-stage continuous N-sufficiency (OCNS), N-deprivation (OCND), N-limitation (OCNL), and also two-stage batch N-starvation (TBNS). Amongst four tested nitrogen manipulation strategies, OCNS performed remarkable promoting effect on cell metabolic growth and the maximum biomass was achieved by 7.39 g/L. Whereas TBNS regime induced the highest lipid content (over 50.5%). Only OCNL treatment augmented the lipid productivity by 232.37 mg/L/day, representing 1.25-fold more than TBNS and even as much as 5.06-fold more than that of OCND strategy. OCNL also strengthened the proportions of saturated (C16:0 and C18:0) and monounsaturated fatty acid (C18:1) which were inclined to high-quality biofuels-making. This might be due to that most part of energy and metabolic flux (e.g. acetyl-CoA) derived from TCA cycle and glycolysis flowed into fatty acids biosynthesis pathway (especially C18:1) response to OCNL manipulation. This study represented a pioneering work of utilizing OCNL for lipids production by C. subellipsoidea and clearly implied that OCNL might be a feasible way for algal lipid production on a commercial scale and also promoted the potential of C. subellipsoidea as an ideal biodiesel feedstock.

The boosted biomass and lipid accumulation in Chlorella vulgaris by supplementation of synthetic phytohormone analogs.

This study attempted at maximizing biomass and lipid accumulation in Chlorella vulgaris by supplementation of natural abscisic acid (ABA) or synthetic 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthaleneacetic acid (NAA) hormone analogs. Amongst three tested additives, NAA-treatment performed remarkable promoting effect on cell growth and lipid biosynthesis. The favorable lipid productivity (418.6mg/L/d) of NAA-treated cells showed 1.48 and 2.24 times more than that of 2,4-D and ABA. NAA-treatment also positively modified the proportions of saturated (C16:0 and C18:0) and monounsaturated fatty acids (C18:1) which were prone to high-quality biofuels-making. Further, NAA-treatment manipulated endogenous phytohormones metabolism leading to the elevated levels of indole-3-acetic acid, jasmonic acid, and salicylic acid and such hormone accumulation might be indispensable for signal transduction in regulating cell growth and lipid biosynthesis in microalgae. In addition, the economic-feasibility and eco-friendly estimation of NAA additive indicated the higher possibilities in developing affordable and scalable microalgal lipids for biofuels.

The boosted lipid accumulation in microalga Chlorella vulgaris by a heterotrophy and nutrition-limitation transition cultivation regime.

A model of heterotrophy and nutrition-limitation transition cultivation for efficient algal biomass and lipid production was proposed in this study, wherein sufficient robust heterotrophic-seed cells submitted into nitrogen-starvation induction for boosted lipid accumulation. The results demonstrated that heterotrophic-seed (HS) achieved specific growth rate of 1.35 day-1 and biomass productivity of 1.93 mg/L/d, representing 6.42- and 32.16-fold, 2.01- and 2.75-fold more than that of photoautotrophic-seed (PS) and mixtrophic-seed (MS). Even though subsequent nutrition-limitation cultivation repressed the growth of HS, the overall lipid productivity caused by nitrogen-starvation was not offset by biomass loss. The most favorable lipid productivity (465.61 mg/L/d) of HS was 3.25 and 52.31 times higher than that of MS and PS. The high content of monounsaturated fatty acids (50.13%) over saturated and polyunsaturated fatty acids (totally 47.39%) in HS cells could provide superior oxidation stability and lower viscosity for biofuels generated from algal biomass feedstock. These findings suggested the feasibility of using heterotrophy and nutrition-limitation transition cultivation for enhancing the overall lipid productivity. Further, several critical enzymes (i.e. G3PDH, ME, and ACAD) were highly related to lipid accumulation and showed especially pronounced up-regulation or down-regulation expression in HS, which provide indications for shedding light on the molecular mechanisms of lipid accumulation and a prospective metabolic engineering for lipid production.

Dendritic cells enhance the activity of human MUC1-stimulated mononuclear cells against breast cancer.

Dendritic cells (DCs) are among the most potent antigen-presenting cells (APCs), stimulating peripheral blood mononuclear cells (PBMCs) to generate antigen-specific cytotoxic T lymphocytes (CTLs). The objectives of this study were to determine if interleukin (IL)-4 is beneficial or detrimental for the generation of human DCs in vitro and to understand whether DCs generated in vitro in the presence or absence of IL-4 stimulate the killing of adenocarcinoma cells by CTLs in vivo. Mucin 1 (MUC1), a glycoprotein found on the surface of adenocarcinoma cells was used to load DCs. MUC1-loaded DCs generated in the absence of IL-4 were superior to their counterparts produced with IL-4 in stimulating PBMCs to kill human breast cancer MCF-7 cells in vitro. A corollary in vivo protection experiment was performed by injecting immunodeficient NOD-SCID mice with MCF-7 cells s.c. and MUC1-loaded CTLs, PBMCs, or DCs generated in the absence of IL-4, i.p. Mice that received CTLs and MUC1-loaded DCs on days 0, 2, 4, 9, 14 and 19 were completely protected against the development of MCF-7-derived tumors, while other schedules conferred lower protection. Therefore, tumor antigen-loaded DCs enhance the efficacy of adoptive CTL transfer, and should thus be considered for combinatorial immunotherapeutic regimens.

Electrocatalytic hydrogenation of 4-chlorophenol on the glassy carbon electrode modified by composite polypyrrole/palladium film.

A polypyrrole/palladium composite film was prepared on a glassy carbon electrode by the electrochemical deposition method. The palladium particles were uniformly dispersed on a polypyrrole film that was previously electrodeposited on a glassy carbon electrode. By controlling the polymerization process of pyrrole, a highly porous polypyrrole film was obtained; this kind of structure provided more surface areas for depositing palladium particles. The sizes of Pd particles deposited on the porous polypyrrole film are about 15-30 nm. The X-ray photoelectron spectroscopy results showed there was strong interaction between polypyrrole film and palladium particles. This modified electrode showed excellent current efficiency (49.5%) for electrochemical hydrogenation of 4-chlorophenol and the phenol was the sole product. The potential effect on the dechlorination process was also investigated.