Insight into the role of chitosan in rapid recovery and re-stabilization of disintegrated aerobic granular sludge.

The disintegration and instability of aerobic granular sludge (AGS) systems during long-term operation pose significant challenges to its practical implementation, and rapid recovery strategies for disintegrated AGS are gaining more attention. In this study, the recovery and re-stabilization of disintegrated AGS was investigated by adding chitosan to a sequencing batch reactor and simultaneously adjusting the pH to slightly acidic condition. Within 7 days, chitosan addition under slight acidity led to the re-aggregation of disintegrated granules, increasing the average particle size from 166.4 µm to 485.9 µm. Notably, sludge volume indexes at 5 min (SVI5) and 30 min (SVI30) decreased remarkably from 404.6 mL/g and 215.1 mL/g (SVI30/SVI5 = 0.53) to 49.1 mL/g and 47.6 mL/g (SVI30/SVI5 = 0.97), respectively. Subsequent operation for 43 days successfully re-stabilized previous collapsed AGS system, resulting in an average particle size of 750.2 µm. These mature and re-stabilized granules exhibited characteristics of large particle size, excellent settleability, compact structure, and high biomass retention. Furthermore, chitosan facilitated the recovery of COD and nitrogen removal performances within 17-23 days of operation. It effectively facilitated the rapid aggregation of disintegrated granules by charge neutralization and bridging effects under a slightly acidic environment. Moreover, the precipitated chitosan acted as carriers, promoting the adhesion of microorganisms once pH control was discontinued. The results of batch tests and microbial community analysis confirmed that chitosan addition increased sludge retention time, enriching slow-growing microorganisms and enhancing the stability and pollutant removal efficiency of the AGS system.

Rapid start-up of an aerobic granular sludge system for nitrogen and phosphorus removal through seeding chitosan-based sludge aggregates.

This study presents a novel strategy to accelerate the start-up of aerobic granular sludge (AGS) system and ensure the nutrient removal during cultivation. This new method consists of preparing the chitosan-based sludge aggregates outside the reactor and then seeding the reactor with such sludge aggregates. To prepare chitosan-based sludge aggregates, chitosan was dissolved in acetic acid solution acting as a cationic flocculant to bind negatively charged sludge together, and then the dissolved chitosan was in situ precipitated by readjusting pH to form stable sludge aggregates. The chitosan-induced charge neutralization and water-insolubility of chitosan were the two main reasons for the super-rapid formation of chitosan-based sludge aggregates. The as-prepared chitosan-based sludge aggregates had a much lower sludge volume index at 30 min (SVI30) (90.1 mL/g) than the original sludge (SVI30 = 328.0 mL/g). They also had some AGS-like characteristics such as large particle size (1300 µm) and fast settling velocity (23.8 m/h). Consequently, short settling time can be achieved and excessive biomass wash-out can be avoided in the rapid start-up of AGS system with chitosan-based sludge aggregates as inoculant, which was beneficial to accelerating sludge granulation while maintaining nutrient removal. Additionally, the abundances of filamentous bacteria and Candidatus Accumulibacter and the content of extracellular polymeric substances increased during cultivation, which could also contribute to the AGS formation. By seeding chitosan-based sludge aggregates in the anaerobic/oxic sequencing batch reactor, complete granulation was rapidly achieved in 10 days, and good removals of nitrogen and phosphorus was obtained after 14-18 days of cultivation.

Oxygenic denitrification for nitrogen removal with less greenhouse gas emissions: Microbiology and potential applications.

Nitrogen pollution is a worldwide problem and has been extensively treated by canonical denitrification (CDN) process. However, the CDN process generates several issues such as intensive greenhouse gas (GHG) emissions. In the past years, a novel biological nitrogen removal (BNR) process of oxygenic denitrification (O2DN) has been proposed as a promising alternative to the CDN process. The classic denitrification four steps are simplified to three steps by O2DN bacteria without producing and releasing the intermediate nitrous oxide (N2O), a potent GHG. In this article, we summarized the findings in previous literatures as well as our results, including involved microorganisms and metabolic mechanisms, functional genes and microbial detection, kinetics and influencing factors and their potential applications in wastewater treatment. Based on our knowledge and experience, the benefits and limitations of the current O2DN process were analyzed. Since O2DN is a new field in wastewater treatment, more research and application is required, especially the development of integrated processes and the quantitative assessment of the contribution of O2DN process in natural habitats and engineered systems.

Folic acid-functionalized polyethylenimine superparamagnetic iron oxide nanoparticles as theranostic agents for magnetic resonance imaging and PD-L1 siRNA delivery for gastric cancer.

Programmed death ligand-1 (PD-L1), which is highly expressed in gastric cancers, interacts with programmed death-1 (PD-1) on T cells and is involved in T-cell immune resistance. To increase the therapeutic safety and accuracy of PD-1/PD-L1 blockade, RNA interference through targeted gene delivery was performed in our study. We developed folic acid (FA)- and disulfide (SS)-polyethylene glycol (PEG)-conjugated polyethylenimine (PEI) complexed with superparamagnetic iron oxide Fe3O4 nanoparticles (SPIONs) as a siRNA-delivery system for PD-L1 knockdown. The characterization, binding ability, cytotoxicity, transfection efficiency, and cellular internalization of the polyplex were determined. At nitrogen:phosphate (N:P) ratios of 10 or above, the FA-PEG-SS-PEI-SPIONs bound to PD-L1 siRNA to form a polyplex with a diameter of approximately 120 nm. Cell-viability assays showed that the polyplex had minimal cytotoxicity at low N:P ratios. The FA-conjugated polyplex showed higher transfection efficiency and cellular internalization in the folate receptor-overexpressing gastric cancer cell line SGC-7901 than a non-FA-conjugated polyplex. Subsequently, we adopted the targeted FA-PEG-SS-PEI-SPION/siRNA polyplexes at an N:P ratio of 10 for function studies. Cellular magnetic resonance imaging (MRI) showed that the polyplex could also act as a T2-weighted contrast agent for cancer MRI. Furthermore, one of four PD-L1 siRNAs exhibited effective PD-L1 knockdown in PD-L1-overexpressing SGC-7901. To determine the effects of the functionalized polyplex on T-cell function, we established a coculture model of activated T cells and SGC-7901 cells and demonstrated changes in secreted cytokines. Our findings highlight the potential of this class of multifunctional theranostic nanoparticles for effective targeted PD-L1-knockdown therapy and MRI diagnosis in gastric cancers.

Adenoviral vector mediates high expression levels of human lactoferrin in the milk of rabbits.

limitations in current technology for generating transgenic animals, such as the time and the expense, hampered its extensive use in recombinant protein production for therapeutic purpose. In this report, we present a simple and less expensive alternative by directly infusing a recombinant adenovirus vector carrying human lactoferrin cDNA into rabbit mammary glands. The milk serum was collected from the infected mammary gland 48 h post-infection and subjected to a 10% SDS-PAGE and Western blotting. An 80-kDa protein was visualized after viral vector infection. With this method, we obtained a high level of expressed human lactoferrin of up to 2.3 mg/ml in the milk. Taken together, the method is useful for the transient high-level expression recombinant proteins, and the approach established here is probably one of the most economical and efficient ways for large-scale production of recombinant proteins of biopharmaceutical interest.

High-level expression of human lactoferrin in the milk of goats by using replication-defective adenoviral vectors.

The expression of human lactoferrin in the mammary gland is an attractive approach to diminish its current production cost. Previous attempts to produce lactorferrin in the milk of transgenic animals resulted in very high cost and uncertain results. In this paper, we have directly infused replication-defective adenovirus encoding human lactoferrin cDNA into the mammary gland of goats via the teat canal. In this way, we obtained a high level of expressed human lactoferrin up to 2g/L in the milk of goats. The milk serum was collected from the infected mammary gland 48 h post-infection and subjected to a 10% SDS-PAGE and Western blotting. A approximately 80-kDa protein was visualized after viral vector infection. Our results demonstrate that intraductal injection of recombinant replication-defective adenovirus vectors may provide a very useful tool for large-scale production of recombinant proteins of biopharmaceutical interest.