Iron particle-integrated anammox granules in baffled reactor: Enhanced settling property and nitrogen removal performance.

This study evaluates iron particle-integrated anammox granules (IP-IAGs) to enhance wastewater treatment efficiency. The IP-IAGs resulted in notable improvements in settleability and nitrogen removal. The settling velocity of IP-IAGs increased by 17.91 % to 2.92 ± 0.20 cm/s, and the total nitrogen removal efficiency in batch mode improved by 6.82 %. These changes indicate enhanced biological activity for effective treatment. In continuous operation, the IP-IAGs reactor showed no accumulation of nitrite until 40 d, reaching a peak nitrogen removal rate (NRR) of 1.54 kg-N/m3·d and a nitrogen removal efficiency of 82.61 %. Furthermore, a partial nitritation-anammox reactor that treated anaerobic digestion effluent achieved a NRR of 1.41 ± 0.09 kg-N/m3·d, proving the applicability of IP-IAGs in real wastewater conditions. These results underscore the potential of IP-IAGs to enhance the efficiency and stability of anammox-based processes, marking a significant advancement in environmental engineering for wastewater treatment.

Diindolylmethane Inhibits Cadmium-Induced Autophagic Cell Death via Regulation of Oxidative Stress in HEL299 Human Lung Fibroblasts.

Cadmium (Cd), a harmful heavy metal, can lead to various pulmonary diseases, including chronic obstructive pulmonary disease (COPD), by inducing cytotoxicity and disturbing redox homeostasis. The aim of the present study was to investigate Cd-mediated cytotoxicity using human lung fibroblasts and the therapeutic potential of 3,3'-diindolylmethane (DIM). Cadmium significantly reduced the cell viability of human embryonic lung (HEL299) cells accompanied by enhanced oxidative stress as evidenced by the increased expression of autophagy-related proteins such as LC3B and p62. However, treatment with DIM significantly suppressed autophagic cell death in Cd-induced HEL299 fibroblasts. In addition, DIM induced antioxidant enzyme activity and decreased intracellular reactive oxygen species (ROS) levels in Cd-damaged HEL299 cells. This study suggests that DIM effectively suppressed Cd-induced lung fibroblast cell death through the upregulation of antioxidant systems and represents a potential agent for the prevention of various diseases related to Cd exposure.

Achieving stable nitrogen removal performance of mainstream PN-ANAMMOX by combining high-temperature shock for selective recovery of AOB activity.

This paper describes the new concept of the mainstream partial nitritation (PN)-anaerobic ammonium oxidation (ANAMMOX) combined with a high-temperature shock strategy for the selective recovery of ammonia-oxidizing bacteria (AOB) activity. In the preliminary test, the temperature shock condition for PN was optimized (60 °C and > 20 min). Based on this, the implementation strategy in a continuous stirred tank reactor (CSTR) system was studied further, and the polyvinyl alcohol (PVA)/sodium alginate carrier exposure ratio (ER) and dissolved oxygen (DO) concentration were considered as primary variables. The AOB activity was recovered selectively when the ER of the carrier ranged from 20 to 40%, and the DO was higher than 2.3 mg O2/L. This was not the case for nitrite-oxidizing bacteria (NOB) (AOB: 1.17±0.1 gNH4+-N/LCarrier/d, NOB: 0.34±0.1 gNO3--N/LCarrier/d). As a result, the activity of AOB was recovered selectively with a decrease in Nitrospira spp., which was verified by kinetic and microbial analyses for the AOB (KS, DO = 3.89 mgO2/L) and NOB (KS, DO = 1.14 mgO2/L). Eventually, the mainstream PN-ANAMMOX was achieved with a nitrogen removal efficiency of 81.5±3.3% for 95 days. The findings provide insight to establishing a stable mainstream PN-ANAMMOX process using a high-temperature shock strategy.

A comparative study of single- and two-phase anaerobic digestion of food waste under uncontrolled pH conditions.

This study compared single- versus two-phase systems for semi-continuous anaerobic digestion of food waste without pH control at varying organic loading rates (OLRs). The methanogenic reactors of both systems required trace element supplementation for stable operation at 3.0 g VS (volatile solids)/L∙d or higher OLRs. Under trace-element supplemented conditions, both systems achieved stable and efficient performance at OLRs up to 4.0 g VS/L∙d. The two-phase system outperformed the single-phase system at 1.0-4.0 g VS/L∙d OLRs, but it failed at an OLR of 5.0 g VS/L∙d. Meanwhile, the single-phase system maintained the stable performance and reached its maximum methane production at this OLR. These results suggest that a single-phase configuration is more advantageous for robust treatment of food waste without pH control at high organic and hydraulic loads. Hydrogenotrophic methanogens dominated the methanogen community throughout the experiment in both systems. Microbial community structure shifts correlated with reactor operation and performance characteristics.

Sequential Combination of Electro-Fenton and Electrochemical Chlorination Processes for the Treatment of Anaerobically-Digested Food Wastewater.

A two-stage sequential electro-Fenton (E-Fenton) oxidation followed by electrochemical chlorination (EC) was demonstrated to concomitantly treat high concentrations of organic carbon and ammonium nitrogen (NH4+-N) in real anaerobically digested food wastewater (ADFW). The anodic Fenton process caused the rapid mineralization of phenol as a model substrate through the production of hydroxyl radical as the main oxidant. The electrochemical oxidation of NH4+ by a dimensionally stable anode (DSA) resulted in temporal concentration profiles of combined and free chlorine species that were analogous to those during the conventional breakpoint chlorination of NH4+. Together with the minimal production of nitrate, this confirmed that the conversion of NH4+ to nitrogen gas was electrochemically achievable. The monitoring of treatment performance with varying key parameters (e.g., current density, H2O2 feeding rate, pH, NaCl loading, and DSA type) led to the optimization of two component systems. The comparative evaluation of two sequentially combined systems (i.e., the E-Fenton-EC system versus the EC-E-Fenton system) using the mixture of phenol and NH4+ under the predetermined optimal conditions suggested the superiority of the E-Fenton-EC system in terms of treatment efficiency and energy consumption. Finally, the sequential E-Fenton-EC process effectively mineralized organic carbon and decomposed NH4+-N in the real ADFW without external supply of NaCl.

Use of Swine Wastewater as Alternative Substrate for Mycelial Bioconversion of White Rot Fungi.

Seven white rot fungal species were tested for growth as mycelia using swine wastewater (SW), an agro-waste with tremendous environmental footprint, as the sole nutrient source. The SW contained high concentrations of carbon and nitrogen components, which could support nutritional requirements for mycelial growth. Out of the seven species, Pleurotus ostreatus and Hericium erinaceus were successfully cultivated on the SW medium using solid-state fermentation. Response surface methodology was employed to determine the combination of pH, temperature (T), and substrate concentration (C) that maximizes mycelial growth rate (Kr) for the two species. The optimum condition was estimated as pH = 5.8, T = 28.8 °C, and C = 11.2 g chemical oxygen demand (COD)/L for P. ostreatus to yield Kr of 11.0 mm/day, whereas the greatest Kr (3.1 mm/day) was anticipated at pH = 4.6, T = 25.5 °C, and C = 11.9 g COD/L for H. erinaceus. These Kr values were comparable to growth rates obtained using other substrates in the literature. These results demonstrate that SW can be used as an effective substrate for mycelial cultivation of the two white rot fungal species, suggesting an alternative method to manage SW with the production of potentially valuable biomass.

Abietic acid has an anti-obesity effect in mice fed a high-fat diet.

We investigated the anti-obesity effect of abietic acid in mice fed a high-fat diet with emphasis on changes in adipogenesis in epididymal adipose tissues. Male C57BL/6J mice were divided into four groups and fed a normal diet, a high-fat diet (HFD), or HFD plus oral administration of abietic acid (20 mg/kg of body weight/day [LA] or 40 mg/kg of body weight/day [HA]) for 8 weeks. Compared with the HFD group, mice orally administered 40 mg of abietic acid/kg of body weight/day exhibited significantly decreased body weight and adipose tissue weights. Serum triglyceride concentrations in the HA group were significantly lower than those in the HFD group, as were the levels of serum insulin and leptin. Hematoxylin and eosin staining revealed that epididymal adipose tissue mass was decreased by abietic acid administration. Abietic acid also inhibited the protein expression of sterol regulatory element-binding protein-1c, CCAAT/enhancer-binding protein α, and CD36 in epididymal adipose tissues, which are up-regulated by HFDs. These data demonstrate that abietic acid has an anti-obesity effect in mice mediated by the regulation of adipogenesis.

Methanogenic community shift in anaerobic batch digesters treating swine wastewater.

Qualitative and quantitative molecular analysis techniques were used to determine associations between differences in methanogenic microbial communities and the efficiency of batch anaerobic digesters. Two bioreactors were initially seeded with anaerobic sludge originating from a local municipal wastewater treatment plant and then supplemented with swine wastewater. Differences were observed in the total amount of methane produced in the two bioreactors (7.9L/L, and 4.5L/L, respectively). To explain these differences, efforts were taken to characterize the microbial populations present using a PCR-based DGGE analysis with methanogenic primer and probe sets. The groups Methanomicrobiales (MMB), Methanobacteriales (MBT), and Methanosarcinales (MSL) were detected, but Methanococcales (MCC) was not detected. Following this qualitative assay, real-time PCR was used to investigate quantitative differences in the populations of these methanogenic orders. MMB was found to be the dominant order present and its abundance patterns were different in the two digesters. The population profiles of the other methanogenic groups also differed. Through redundancy analysis, correlations between the concentrations of the different microbes and chemical properties such as volatile fatty acids were calculated. Correlations between MBT and MSL populations and chemical properties were found to be consistent in both digesters, however, differences were observed in the correlations between MMB and propionate. These results suggest that interactions between populations of MMB and other methanogens affected the final methane yield, despite MMB remaining the dominant group overall. The exact details of why changes in the MMB community caused different profiles of methane production could not be ascertained. However, this research provides evidence that microbial behavior is important for regulating the performance of anaerobic processes.

Qualitative and quantitative assessment of microbial community in batch anaerobic digestion of secondary sludge.

Microbial community shifts were determined by denaturing gradient gel electrophoresis (DGGE) and real-time PCR for an anaerobic batch digester treating secondary sludge. The batch process was successfully operated with an organic removal efficiency of 35% associated with a 91% decrease in the bacterial 16S rRNA gene concentration. The microbial community structures showed continuous shifts within four bacterial phyla and three archaeal orders. Several bacterial species, such as Fusibacter-related, Clostridium-like, and Syntrophus-like organisms, appeared to be responsible for acidogenesis or syntrophic acid degradation. Both hydrogenotrophic and aceticlastic methanogens appear to have been involved in the methanogenesis with the acidogenic products. The quantitative structure of the methanogenic populations varied continuously, with the growth of Methanomicrobiales and Methanosarcinales in series, to result in a Methanomicrobiales-dominant population. The ordination of microbial community structures demonstrated that the quantitative methanogenic structure converged to the seed inoculum while the bacterial and archaeal DGGE band patterns diverged. These results provide an insight into the microbial behavior in the transitional phase (e.g., a start-up period) of anaerobic sludge digestion.

Effect of microwave irradiation on cellular disintegration of Gram positive and negative cells.

This research investigated the effect of microwave irradiation (MWI) on cell disintegration in municipal secondary sludge (MSS). A representative MSS Gram-positive bacterium (Bacillus subtilis) and Gram-negative bacteria (Acinetobacter calcoaceticus and Pseudomonas aeruginosa) were pure cultured separately and treated using MWI. Compared to untreated controls, MWI significantly increased the soluble chemical oxygen demand (COD) (1.8-4.0-fold), soluble protein concentration (1.1-1.8-fold), and soluble carbohydrate concentration (3.2-14.1-fold), with greater increase in the Gram-negative bacteria. After MSS was MWI-treated with different irradiation times, from 0 to 9 min, soluble COD increased gradually from 0.14 to 2.38 g/L (i.e., 72-fold). Effective disintegration of Gram-negative cell walls and of MSS by MWI was confirmed by scanning electron microscopy. These findings suggest that MWI could be an effective pretreatment method for MSS that is dominated by Gram-negative microorganisms.

Effects of prolonged starvation on methanogenic population dynamics in anaerobic digestion of swine wastewater.

This study investigated the relationship between the processes and microbial populations induced by long-term starvation. To demonstrate the effects of starvation, a laboratory-scale anaerobic reactor was operated in three phases (first reaction, starvation, second reaction) for 316 days. During the first reaction, the chemical oxygen demand (COD) concentration decreased by about 70% of the input swine wastewater and 64L of methane gas was produced; during the second reaction, there was a 63% COD reduction and 36L of methane was produced. The methanogenic diversity, qualitatively monitored with denaturing gradient gel electrophoresis using archaeal 16S rRNA gene primers, was not different between two reactions. However, DNA copy numbers of Methanosarcinales, quantitatively monitored with quantitative real-time polymerase chain reaction using order-level 16S rRNA gene primers, showed the changed results. Cell numbers of Methanosarcinales and methanogenic activity were important factors determining the different efficiencies of the process.

Effect of high temperature on bacterial community dynamics in anaerobic acidogenesis using mesophilic sludge inoculum.

In this study, we investigated the microbial community dynamics in thermal acidogenesis using mesophilic sludge. From the result of optimization with a response surface methodology, the acidogenic optimum conditions predicted were a hydraulic retention time of 2.0 days and 51 degrees C. Denaturing gradient gel electrophoresis (DGGE) profiles shows that the monitored bacterial community present consists of Pseudomonas mendocina, Bacillus halodurans, Clostridium hastiforme, Gracilibacter thermotolerans, and Thermomonas haemolytica. Among these, B. halodurans, G. thermotolerans, and T. haemolytica are reported to ferment carbohydrates thermotolerantly. In contrast, P. mendocina disappeared in the acidogenesis process because of its mesophilicity. In addition, C. hastiforme, G. thermotolerans originating from mesophilic anaerobic sludge were detected in the thermal acidogenesis. Based on this finding, we inferred that most thermophiles detected as DGGE bands could grow catalyzing carbohydrates metabolism in swine wastewater to produce volatile fatty acids thermotolerantly.

Fermentation and growth kinetic study of Aeromonas caviae under anaerobic conditions.

Although Aeromonas caviae is pathogenic to a broad range of invertebrates including human, frequent in aquatic environments, and potentially vital for acidogenesis in anaerobic digestion, virtually no biokinetic information on its anaerobic growth is at hand. Therefore, this study focused on evaluating its anaerobic growth kinetics on glucose. To provide a set of relevant biokinetic coefficients for modeling, a combination of curve fitting and numerical modeling was used. Microcultivations were performed at eight different initial glucose concentrations of 0.1 to 2.5 g l(-1) to establish a function of specific growth rate versus substrate concentration. A batch anaerobic bioreactor was then operated to collect a data set for the numerical analysis. Kinetic coefficients were estimated from three different biomass growth profiles monitored by optical density, volatile suspended solids (VSS), or DNA measurement, and applied for simulating continuous operations at various hydraulic retention times (HRTs). Assuming the influent glucose concentration is 5,000 mg l(-1), the substrate utilization efficiency predicted to be 77.2% to 92.0% at 17 to 36 h HRTs. For the VSS-model-based simulation, the washout HRT was estimated to be 16.6 h, and similar for the other models. Overall, the anaerobic biokinetic coefficients of A. caviae grown on glucose were successfully estimated and found to follow a substrate inhibition model.

Quantitative analysis of methanogenic community dynamics in three anaerobic batch digesters treating different wastewaters.

Quantitative changes in methanogenic community structures, associated with performance data, were investigated in three anaerobic batch digesters treating synthetic glucose medium, whey permeate, and liquefied sewage sludge. All digesters were initially seeded with anaerobic sludge obtained from a local municipal wastewater treatment plant. Dynamics of methanogenic populations were monitored, at order and family levels, using real-time PCR based on the 16S rRNA gene. The molecular monitoring revealed that, in each digester, the quantitative structure of methanogenic community varied continuously over treatment time and the variation corresponded well to the changes in chemical profiles. Biphasic production of methane, associated with successive increases in aceticlastic (mainly Methanosarcinaceae) and hydrogenotrophic (mainly Methanomicrobiales) methanogenic groups, was observed in each digester. This corresponded to the diauxic utilization of acetate and longer-chain volatile fatty acids (C(3)-C(6)), mainly propionate. Additionally, the non-metric multidimensional scaling (NMDS) analysis of the quantification results demonstrated that the community shift patterns in three digesters were totally different from each other. Considering that the operating conditions in all trials were identical except substrates, the differences in quantitative shift profiles were suggested to be due to the different substrate compositions. This implied that the composition of wastewater could affect the evolution of quantitative methanogenic community structure in an anaerobic process. Overall, our results suggested that more attention to quantitative as well as qualitative approaches on microbial communities is needed for fundamental understanding of anaerobic processes, particularly under dynamic or transitional conditions.

Use of order-specific primers to investigate the methanogenic diversity in acetate enrichment system.

The applicability of order-specific primers in minimizing the possible underestimation of microbial diversity was evaluated via denaturing gradient gel electrophoresis (DGGE) analysis of a lab-scale anaerobic digester. Initially, a population analysis with real-time quantitative PCR demonstrated the existence of three methanogenic orders--Methanobacteriales, Methanomicrobiales, and Methanosarcinales--throughout the reaction period. DGGE analyses with three pairs of order-specific primers yielded eight operational taxonomic units (OTUs), whereas DGGE analysis with two independent Archaea-specific primers identified only five. Moreover, the order-specific primers amplified at least one OTU affiliated with each order, whereas no members of Methanobacteriales or Methanomicrobiales were identified with Archaea-specific primers in most samples. These findings provide evidence that order-specific analysis can detect the diversity of methanogens in greater detail than conventional Archaea-specific analysis.

Methanogenic profiles by denaturing gradient gel electrophoresis using order-specific primers in anaerobic sludge digestion.

In the present study, the diversity of methanogenic populations was monitored for 25 days, together with the process data for an anaerobic batch reactor treating waste-activated sludge. To understand this microbial diversity and dynamics, 16S rRNA-gene-targeted denaturing gradient gel electrophoresis (DGGE) fingerprinting was conducted at two different taxonomic levels: the domain and order levels. The DGGE profiles of the domain Archaea and the three orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales were comparatively analyzed after each DGGE band was sequenced to enable identification. The DGGE profiles of the three orders showed methanogens belonging to each order that were not detected in the DGGE profile of the Archaea. This discrepancy may have resulted from PCR bias or differences in the abundances of the three microbial orders in the anaerobic bioreactor. In conclusion, to fully understand the detailed methanogenic diversity and dynamics in an anaerobic bioreactor, it is necessary to conduct DGGE analysis with 16S rRNA gene primers that target lower taxonomic groups.