Variations in antibiotic resistance genes and microbial community in sludges passing through biological nutrient removal and anaerobic digestion processes in municipal wastewater treatment plants.

Antimicrobial resistance (AMR) represents a relentless, silent pandemic. Contributing to this are wastewater treatment plants (WWTPs), a potential source of antibiotic resistance genes' (ARGs) transmission to the environment, threatening public health. The presence of ARGs in pathogenic bacteria and their release into the environment by WWTPs threatens the public health. The current study investigated changes in ARGs' abundance in biological nutrient removal (BNR) processes and anaerobic digestion (AD) reactors of two WWTPs. Also, microbial community structure, which is known to shape the distribution and abundance of ARGs, was also analyzed. The relative abundance of eight ARGs (tetX, tetA, tetM, TEM, sul1, sul2, ermB and qnrD) was quantified as ARGs' copies/16 S rRNA gene copies using quantitative polymerase chain reaction (qPCR). Microbial community composition was assessed by 16 S rRNA microbiome sequencing analysis. TetX was prevalent among the eight ARGs, followed by TEM and sul1. However, its abundance was decreased in the AD sludges compared to BNR sludges. Proteobacteria was the major bacterial phylum found in all the sludge samples, while Arcobacter, 12up and Acidovorax were the predominant genera. Acinetobacter and Flavobacterium were significantly more abundant in the BNR sludges, while 12up and Aeromonas were predominant in AD sludges. Principal component analysis (PCA) revealed a clear difference in dominant ARGs and bacteria between the sludges in the processes of BNR and AD of the two WWTPs. Clinically relevant bacterial genera, Klebsiella and Enterococcus, found in both the BNR and AD sludges, were significantly correlated with the tetX gene. Throughout this study, the relationship between microbial communities and specific ARGs was revealed, illustrating that the composition of the microbial community could play a vital role in the abundance of ARGs. These results will better inform future studies aimed at controlling the spread of ARGs and their potential hosts from WWTPs.

Selective strontium adsorption using synthesized sodium titanate in aqueous solution.

Amorphous sodium titanates were synthesized using a mid-temperature sol-gel method for evaluation as selective adsorbents of strontium in the presence of cesium or metal cations (Al3+, Mg2+, Ca2+, and Mn2+) from aqueous solution. Synthesized sodium titanate showed high adsorption capacity and selectivity for strontium. The maximum adsorption capacity of strontium by sodium titanate was 193.93 mg g-1 in aqueous solution containing an initial concentration of 5 mM (438.60 mg L-1) strontium and 5 mM (666.67 mg L-1) cesium, and this sodium titanate removed 99.9% of the strontium and 40.67% of cesium from an aqueous solution that had an initial concentration of 1.14 mM (100 mg L-1) strontium and 0.75 mM (100 mg L-1) cesium. Strontium adsorption by synthesized sodium titanate followed pseudo-second-order kinetics and a generalized Langmuir isotherm model, and reached an adsorption equilibrium within 1 h with high adsorption capacity at equilibrium. Adsorbed strontium onto synthesized sodium titanate showed the behavior of forming a strontium titanate structure with a titanate frame via surface precipitation.

Effects of biological pretreatments of microalgae on hydrolysis, biomethane potential and microbial community.

Parameters of temperature-phased anaerobic digestion (TPAD) were varied to study their effects on hydrolysis, biomethane potential (BMP), and microbial diversity of microalgae biodegradation. Anaerobic pretreatments at 85 °C demonstrated the release of soluble carbohydrate and protein molecules under low microbial metabolic activity. However, at 55 °C, anaerobic pretreatments showed superior performance in methane yield, nutrient release, and volatile fatty acids (VFAs) production due to dominant Clostridium. Furthermore, the highest destruction of volatile solids (VS) was observed during aerobic pretreatments at 55 °C under the influence of various quantities of these genera - Luteimonas, Symbiobacterium, Soehngenia, Thermobacillus, and Ureibacillus. Statistical analysis revealed that hydrolysis and BMP were not correlated. However, soluble nitrogen and phosphorous showed strong correlation with methane (r = 0.623 and 0.948, respectively) under thermo-anaerobic pretreatment, while VS removal and concentrations of acetic and butyric acids and lipids were positively correlated with each other under thermo-aerobic pretreatment.

Changes in microbial communities during volatile fatty acid production from cyanobacterial biomass harvested from a cyanobacterial bloom in a river.

Volatile fatty acid (VFA) production, utilization of soluble organic compounds, and associated microbial consortia were investigated after different pretreatments (untreated, alkaline, and thermal-alkaline) using cyanobacterial biomass as a substrate. Compared to the untreated control, soluble carbohydrate concentrations were almost the same after alkaline and thermal-alkaline pretreatments, but soluble protein concentration was 1.58 times higher after alkaline pretreatment and 1.81 times higher after thermal-alkaline pretreatment. However, the highest degree of acidification was obtained after alkaline pretreatment (55.36 ±â€¯3.00%). Microbial communities in the untreated control differed only slightly from those after thermal-alkaline pretreatment, but were clearly distinct from those after alkaline pretreatment. After alkaline pretreatment, protein-utilizing bacteria became relatively predominant. These results revealed the relationships between efficiency of VFA production and the shift in microbial community.

Biodiesel production by various oleaginous microorganisms from organic wastes.

Biodiesel is a biodegradable and renewable fuel. A large amount of research has considered microbial oil production using oleaginous microorganisms, but the commercialization of microbial lipids produced in this way remains uncertain due to the high cost of feedstock or low lipid yield. Microbial lipids can be typically produced by microalgae, yeasts, and bacteria; the lipid yields of these microorganisms can be improved by using sufficient concentrations of organic carbon sources. Therefore, combining low-cost organic compounds contained in organic wastes with cultivation of oleaginous microorganisms can be a promising approach to obtain commercial viability. However, to achieve effective bioconversion of low-cost substrates to microbial lipids, the characteristics of each microorganism and each substrate should be considered simultaneously. This article discusses recent approaches to developing cost-effective microbial lipid production processes that use various oleaginous microorganisms and organic wastes.

Volatile fatty acid recovery by anaerobic fermentation from blue-green algae: Effect of pretreatment.

The aims of this study were to quantify how pretreatment affects production of volatile fatty acids (VFAs) from cyanobacterial biomass and production of subsequent microbial lipid by an oleaginous microorganism that uses the VFAs as carbon sources. The highest biomass solubilization was obtained using thermal-alkaline (th-alkaline) pretreatment (33.1%), followed by alkaline pretreatment (29.1%), and thermal pretreatment (7.2%), but the highest VFA yield was obtained using alkaline pretreatment (0.54±0.02g/gVS), followed by the untreated condition (0.47±0.03g/gVS), and th-alkaline pretreatment (0.44±0.02g/gVS). Although VFA yield was higher using alkaline pretreatment condition than in the untreated condition, the difference was not great. However, lipid productivity by Cryptococcus curvatus after the alkaline pretreatment condition was 2.0-fold higher than that under the untreated condition. This study confirmed the feasibility of using biologically produced VFAs from cyanobacterial biomass for microbial lipid production by the oleaginous microorganism.

Enhanced microalgal biomass and lipid production from a consortium of indigenous microalgae and bacteria present in municipal wastewater under gradually mixotrophic culture conditions.

The goal of this study was to investigate the influences of gradually mixotrophic culture conditions on microalgal biomass and lipid production by a consortium of indigenous microalgae and bacteria present in raw municipal wastewater. Lab-scale photobioreactors containing the consortium were operated in repeated batch mode. Initial cultivation (phase I) was performed using only the municipal wastewater, then 10% and 25% of the reactor volumes were replaced with the effluent from a sewage sludge fermentation system producing volatile fatty acids (SSFV) at the beginnings of phase II and phase III, respectively. The highest biomass productivity (117.1±2.7mg/L/d) was attained during phase II, but the lipid productivity (17.2±0.2mg/L/d) was attained during phase III. The increase in the effluent from the SSFV influenced microalgal diversity with a preference for Chlorella sp., but bacterial diversity increased significantly during phase III.

Influence of temperature on volatile fatty acid production and microbial community structure during anaerobic fermentation of microalgae.

The objectives of this study were to investigate the effect of operating temperature on volatile fatty acids (VFAs) production from microalgal biomass, and to clarify the relationships between VFAs accumulation at different temperatures and the corresponding bacterial communities. The VFA yields were 0.10±0.017, 0.12±0.008, and 0.34±0.009 g/g VS at 35, 45, and 55 °C, respectively. The proportion of acetic acid decreased from 85.6% to 65.8% as operating temperature increased, whereas that of propionic acid increased from near 0% to 15.5% and that of iso-valeric acid remained relatively stable (10.2-11.2%). Bacterial communities at different operating temperatures consisted mostly of the phyla Proteobacteria, Bacteroidetes, and Firmicutes, which can degrade organic compounds effectively. Bacillus sp. was more predominant at 55 °C than at mesophilic temperatures, suggesting that this microorganism contributed significantly to the higher hydrolysis rate and VFA yield at this operating temperature.

Effects of pH control and concentration on microbial oil production from Chlorella vulgaris cultivated in the effluent of a low-cost organic waste fermentation system producing volatile fatty acids.

The objective of this study was to investigate the feasibility of applying volatile fatty acids (VFAs) produced from low-cost organic waste to the major carbon sources of microalgae cultivation for highly efficient biofuel production. An integrated process that consists of a sewage sludge fermentation system producing VFAs (SSFV) and mixotrophic cultivation of Chlorella vulgaris (C. vulgaris) was operated to produce microbial lipids economically. The effluents from the SSFV diluted to different concentrations at the level of 100%, 50%, and 15% were prepared for the C. vulgaris cultivation and the highest biomass productivity (433±11.9 mg/L/d) was achieved in the 100% culture controlling pH at 7.0. The harvested biomass included lipid contents ranging from 12.87% to 20.01% under the three different effluent concentrations with and without pH control. The composition of fatty acids from C. vulgaris grown on the effluents from the SSFV complied with the requirements of high-quality biodiesel. These results demonstrated that VFAs produced from the SSFV are favorable carbon sources for cultivating C. vulgaris.

Biodiesel production from Scenedesmus bijuga grown in anaerobically digested food wastewater effluent.

Microalgae, Scenedesmus bijuga, was cultivated in anaerobically digested food wastewater effluent (FWE) to treat the wastewater and produce biodiesel simultaneously. Three different mixing ratios with municipal wastewater were compared for finding out proper dilution ratio in biodiesel production. Of these, 1/20 diluted FWE showed the highest biomass production (1.49 g/L). Lipid content was highest in 1/10 diluted FWE (35.06%), and the lipid productivity showed maximum value in 1/20 diluted FWE (15.59 mg/L/d). Nutrient removal was also measured in the cultivation. FAME compositions were mainly composed of C16-C18 (Over 98.94%) in S. bijuga. In addition, quality of FAMEs was evaluated by Cetane Number (CN) and Bis-allylic Position Equivalent (BAPE).

Characterization of membrane foulants in a pilot-scale tunnel construction wastewater treatment process.

A pilot-scale combination of a microfiltration (MF) and a reverse osmosis (RO) membrane system was applied on-site to treat tunnel construction wastewater. The MF membrane system initially removed contaminants (turbidity of less than 0.3 NTU) in the form of particulate materials in the feed water, thereby allowing the combined MF-RO system to efficiently remove more than 99% of known organic and inorganic contaminants and qualify the reclaimed water for reuse. The MF membrane autopsy analysis using X-ray fluorescence (XRF) and inductively coupled plasma-mass spectrometry (ICP-MS) revealed that the dominant foulants were inorganic deposits involving Si, Al and Fe, comprising the main components of cement materials, as well as deposits involving heavy metals such as Mn, Cu and Zn in the form of particles. Thus, thick cake contaminants shown by field emission scanning electron microscope (FE-SEM) images might be induced via suspended solids consisting of cement and clay materials and metals.

Influence of thermophilic aerobic digestion as a sludge pre-treatment and solids retention time of mesophilic anaerobic digestion on the methane production, sludge digestion and microbial communities in a sequential digestion process.

In this study, the changes in sludge reduction, methane production and microbial community structures in a process involving two-stage thermophilic aerobic digestion (TAD) and mesophilic anaerobic digestion (MAD) under different solid retention times (SRTs) between 10 and 40 days were investigated. The TAD reactor (RTAD) was operated with a 1-day SRT and the MAD reactor (RMAD) was operated at three different SRTs: 39, 19 and 9 days. For a comparison, control MAD (RCONTROL) was operated at three different SRTs of 40, 20 and 10 days. Our results reveal that the sequential TAD-MAD process has about 42% higher methane production rate (MPR) and 15% higher TCOD removal than those of RCONTROL when the SRT decreased from 40 to 20 days. Denaturing gradient gel electrophoresis (DGGE) and real-time PCR results indicate that RMAD maintained a more diverse bacteria and archaea population compared to RCONTROL, due to the application of the biological TAD pre-treatment process. In RTAD, Ureibacillus thermophiles and Bacterium thermus were the major contributors to the increase in soluble organic matter. In contrast, Methanosaeta concilii, a strictly aceticlastic methanogen, showed the highest population during the operation of overall SRTs in RMAD. Interestingly, as the SRT decreased to 20 days, syntrophic VFA oxidizing bacteria, Clostridium ultunense sp., and a hydrogenotrophic methanogen, Methanobacterium beijingense were detected in RMAD and RCONTROL. Meanwhile, the proportion of archaea to total microbe in RMAD and RCONTROL shows highest values of 10.5 and 6.5% at 20-d SRT operation, respectively. Collectively, these results demonstrate that the increased COD removal and methane production at different SRTs in RMAD might be attributed to the increased synergism among microbial species by improving the hydrolysis of the rate limiting step in sludge with the help of the biological TAD pre-treatment.

An innovative sewage sludge reduction by using a combined mesophilic anaerobic and thermophilic aerobic process with thermal-alkaline treatment and sludge recirculation.

Lab-scale High Efficiency Digestion (HED) systems containing a Mesophilic Anaerobic Reactor (MAR), Thermophilic Aerobic Reactor (TAR), liquid/solid separation unit, and thermal-alkaline treatment were developed to evaluate the efficiencies of sludge reduction and methane production. The HED process was divided into three phases to examine the influence of sludge pretreatment and pretreated sludge recirculation using TCOD and VSS reduction, COD solubilization, and methane production. The VSS removal with a solid/liquid separation unit, sludge recirculation, and thermal-alkaline treatment drastically increased up to 95% compared to the feed concentration. In addition, the results of COD solubilization and VSS/TSS showed that the solubilization of cells and organic matters by the thermal-alkaline treatment was highly increased, which was also consistent with the SEM images. In particular, the methane production rate increased 24-fold when the feed sludge and recirculated sludge were pretreated together. Collectively, the HED experiments performed with sludge recirculation and thermal-alkaline treatment demonstrated that the HED systems can be successfully employed for highly efficient sewage sludge reduction and methane gas production.

Influence of operational parameters on nitrogen removal efficiency and microbial communities in a full-scale activated sludge process.

To improve the efficiency of total nitrogen (TN) removal, solid retention time (SRT) and internal recycling ratio controls were selected as operating parameters in a full-scale activated sludge process treating high strength industrial wastewater. Increased biomass concentration via SRT control enhanced TN removal. Also, decreasing the internal recycling ratio restored the nitrification process, which had been inhibited by phenol shock loading. Therefore, physiological alteration of the bacterial populations by application of specific operational strategies may stabilize the activated sludge process. Additionally, two dominant ammonia oxidizing bacteria (AOB) populations, Nitrosomonas europaea and Nitrosomonas nitrosa, were observed in all samples with no change in the community composition of AOB. In a nitrification tank, it was observed that the Nitrobacter populations consistently exceeded those of the Nitrospira within the nitrite oxidizing bacteria (NOB) community. Through using quantitative real-time PCR (qPCR), nirS, the nitrite reducing functional gene, was observed to predominate in the activated sludge of an anoxic tank, whereas there was the least amount of the narG gene, the nitrate reducing functional gene.

Response of nitrifying bacterial communities to the increased thiocyanate concentration in pre-denitrification process.

Changes in process performance and the nitrifying bacterial community associated with an increase of thiocyanate (SCN-) loading were investigated in a pre-denitrification process treating industrial wastewater. The increased SCN- loading led to the concentration of total nitrogen (TN) in the final effluent, but increasing the internal recycling ratio as an operation parameter from 2 to 5 resulted in a 21% increase in TN removal efficiency. In the aerobic reactor, we found that the Nitrosomonas europaea lineage was the predominant ammonia oxidizing bacteria (AOB) and the percentages of the AOB population within the total bacteria increased from about 4.0% to 17% with increased SCN- concentration. The increase of nitrite loading seemed to change the balance between Nitrospira and Nitrobacter, resulting in the high dominance of Nitrospira over Nitrobacter. Meanwhile, a Thiobacillus thioparus was suggested to be the main microorganism responsible for the SCN- biodegradation observed in the system.