Antibiotics and antibiotic resistance genes removal in biological aerated filter.

Two laboratory-level biological aerated filters (BAF) were constructed to explore their treatment capacity for simulated antibiotic wastewater at high (1 - 16 mg/L) and low (0 - 0.5 mg/L) concentrations. Results showed that BAF was capable of removing both sulfonamides and tetracyclines with an efficiency of over 90 % at 16 mg/L. The main mechanism for removing antibiotics was found to be biodegradation followed by adsorption. Paenarthrobacter was identified as the key genus in sulfonamides degradation, while Hydrogenophaga played a crucial role in tetracyclines degradation. Antibiotics resistant genes such as intI1, sul1, sul2, tetA, tetW and tetX were frequently detected in the effluent, with interception rates ranging from 105 - 106 copies/mL. The dominated microorganisms obtained in the study could potentially be utilized to enhance the capacity of biological processes for treating antibiotics contaminated wastewater. These findings contribute to a better understanding of BAF treating wastewater containing antibiotics and resistant genes.

Nitrogen removal of thermal hydrolysis-anaerobic digestion liquid: A review.

Thermal hydrolysis (TH) pretreatment, as an anaerobic digestion (AD) pretreatment, has not only been verified in the laboratory but also frequently employed in actual engineering. However, the properties of anaerobic digestion liquid (ADL), such as high organic matter concentration, high ammonia nitrogen (NH4+-N) concentration, and low carbon-nitrogen ratio (C/N), have posed some difficulties in the follow-up treatment. To address the above issues, the autotrophic nitrogen removal (ANR) process is developed to treat ADL. Due to the NH4+-N, organic materials, toxic and harmful substances in the ADL that might directly impact the activity of functional bacteria, the ADL should be treated before being fed into the ANR process. This paper provided a focused review of the thermal hydrolysis-anaerobic digestion process (TH-ADP) mechanism and the ANR mechanism, summarized the existing difficulties in the treatment of thermal hydrolysis-anaerobic digestion liquid (TH-ADL), assessed the research status thoroughly, and offered the potential solutions to the problems.

Deciphering the fate of osmotic stress priming on enhanced microorganism acclimation for purified terephthalic acid wastewater treatment with high salinity and organic load.

Osmotic stress priming (OSP) was an effective management strategy for improving microbial acclimation to salt stress. In this study, the interaction between pollutants and microbiota, and microbial osmoregulation were investigated triggered by OSP (alternately increasing salinity and organic loading). Results showed that OSP significantly improved COD removal from 31.53 % to 67.99 % and mitigated the terephthalate inhibition produced by toluate, decreasing from 1908.08 mg/L to 837.16 mg/L compared with direct priming. Due to an increase in salinity, Pelotomaculum and Mesotoga were enriched to facilitate terephthalate degradation and syntrophic acetate oxidation (SAO). And organic load promoted acetate formation through syntrophic metabolism of Syntrophorhabdus/Pelotomaculum and SAO-dependent hydrogenotrophic methanogenesis. K+ absorbing, proline and trehalose synthesis participated in osmoregulation at 0.5 % salinity, while only ectoine alleviated intracellular osmolarity under 1.0 % salinity with OLR of 0.44 kg COD /m3. This study provided in-depth insight for microbial acclimation process of anaerobic priming of saline wastewater.

Temperature-regulated and starvation-induced refractory para-toluic acid anaerobic biotransformation.

Little research was focused on the anerobic degradation of refractory para-toluic acid at present. Thus, temperature-regulated anaerobic system of para-toluic acid fed as sole substrate was built and investigated via microbiota, metabolism intermediates, and function prediction in this study. Results showed that low methane yield was produced in para-toluic acid anaerobic system at alkaline condition. And the causes were owing to anaerobic methane oxidation and potentially H2S production at 37 °C, N2 production by denitrification before starvation and propionic acid occurrence after starvation at 27 °C, and production of N2 and free ammonia, and accumulation of acetic acid at 52 °C. Simultaneously, hydrogenotrophic methanogenesis dependent on syntrophic acetate oxidation (SAO) was predominant, facilitating the removal of para-toluic acid at 52 °C. Moreover, the key intermediate changed from phthalic acid of 37 °C and 27 °C before starvation to terephthalic acid of 52 °C. Starvation promoted removal of para-toluic acid through benzoyl-CoA pathway by Syntrophorhabdus, enrichment of syntrophic propionate degraders of Bacteroidetes and Ignavibacteriaceae, and increase of methylotrophic methanogens.

Simultaneous nitrification and denitrification in membrane bioreactor: Effect of dissolved oxygen.

Membrane bioreactor with the floc activated sludge (mixed liquor suspended solids (MLSS) = 7500 mg/L) was constructed in this work for simultaneously nitrification and denitrification (SND). The effect of dissolved oxygen (DO) on SND process and the nitrogen pathways were investigated. The average TN removal efficiencies were 63.05%, 91.17%, 87.04% and 70.02% for DO 0.5, 1, 2 and 3 mg/L systems, respectively. The effluent ammonia concentration was continuously lower than 5.0 mg/L when the DO was higher than 1 mg/L. Nitrogen in DO 1 and DO 2 mg/L systems was mainly removed via the SND process. The rise of DO concentration increased the abundance of nitrite oxidizing bacteria (NOB) and Nitrospira was the predominant NOB in all the four MBRs. Dechloromonas and Azoarcus were the dominant denitrifying bacteria (DNB) in DO 1 systems responsible for nitrite denitrification. The dominant aerobic DNB Pseudomonas also contributed SND via nitrate denitrification and was little affected by DO changes. Nitrate reductase was the main enzyme for the reduction of NO3--N to NO2--N, and narG was the main responsible gene. Nitrite oxidoreductase was the main enzyme for the oxidation of NO2--N to NO3--N, and nxrA was the main responsible gene in all the four MBR systems.

Enhanced pollutants removal and high-value cell inclusions accumulation with Fe2+ in heavy oil refinery treatment system using Rhodopseudomonas and Pseudomonas.

Metal ions has been widely used as a method of improving pollutant removal efficiency in wastewater biological treatment system. In order to enhance pollutants removal and high-value cell inclusions accumulation in heavy oil refinery wastewater treatment systems using PSB, different reactors were built feeding with different Fe2+ concentrations respectively, and run with enriching Rhodopseudomonas and Pseudomonas in the reactors. Solute chemical oxygen demand (SCOD), ammonia (NH4+-N), nitrate nitrogen (NO3--N), nitrous nitrogen (NO2--N), Fe2+, and related cell inclusions were all detected, moreover, microbial community structure and the quantity of Rhodopseudomonas and Pseudomonas were also detected. The results showed that at the optimal dosage of Fe2+ with 20 mg/L, the corresponding removal ratios of solute chemical oxygen demand and ammonia were 73.51% and 92.26%, respectively. The yields of carotenoid, bacteriochlorophyll, and coenzyme Q10 were 11.18, 6.75, and 9.84 mg/g-DCW respectively. Furthermore, with 20 mg/L Fe2+ dosage, the relative abundance and gene number of Rhodopseudomonas were the highest in the system, which were 91.57% and 1.843 × 106 gene copies/µL, while Fe2+ had no obvious effect on the growth of Pseudomonas. The results showed that adding Fe2+ has improved the removal of pollutants and accumulation of high-value cells inclusions, also provided theoretical guidance for the treatment of heavy oil refinery wastewater using PSB.

The fate and behavior mechanism of antibiotic resistance genes and microbial communities in anaerobic reactors treating oxytetracycline manufacturing wastewater.

In this study, two parallel-operated expanded granular sludge bed (EGSB) reactors, one used to treat oxytetracycline (OTC) manufacturing wastewater with gradual increase of OTC concentration as experimental reactor and the other fed with the same wastewater without OTC as control reactor, were operated to investigate the behavior of antibiotics resistance genes (ARGs) and mobile genetic elements (MGEs) and their possible relationships with bacterial community among influent, sludge and effluent environments. Though the average absolute abundance of ARGs slightly decreased (0.26 - log), the ARGs' relative abundance normalized to 16S-rRNA gene copy numbers showed a significant upward trend in effluent (2 multiples - increase) and the absolute and relative abundances both extremely increased in anaerobic sludge, indicating that anaerobic treatment process cannot reduce ARGs efficiently, inversely can increase the risk of ARGs through the proliferation of antibiotics resistance bacteria (ARB) under the suppression of OTC. MGEs, bacterial communities and OTC concentration mainly impacted the ARGs profiles, which contributed 88.4% to the variation of ARGs. The differences and correlations of hosts in influent, effluent and sludge were further confirmed by network analysis. Overall, this study enhanced the understanding of the prevalence and transfer of ARGs in OTC production effluents during anaerobic treatment.

Optimizing temperature for enhancing waste activated sludge decomposition in lysozyme and rhamnolipid pretreatment system.

This study investigated the influence of temperature on the hydrolysis and decomposition of waste activated sludge (WAS) during the enhanced pretreatment system with lysozyme and rhamnolipid (Ly + RL). Results showed that temperature increasing from 15℃ to 55℃ could obviously enhance the release of soluble organic matters and WAS decomposition degree within the Ly + RL pretreatment system. Compared to the sum of sole Ly and sole RL pretreatment, Ly + RL combined pretreatment system at 45℃ presented best synergistic hydrolysis performance. The decomposition degree of bacteria and archaea reached 47.6% and 88.1%, respectively. Meanwhile, increasing temperature could recede the diversity of microbial community in the system. Gammaproteobacteria, with the relative abundance of 90.7%, occupied the absolute dominant position at 45℃. Furthermore, with the rise of temperature, more volatile fatty acids were harvested after anaerobic fermentation.

Characteristics of microbial community in EGSB system treating with oxytetracycline production wastewater.

In order to realize the efficient and stable operation of anaerobic digestion for oxytetracycline (OTC) production wastewater which contains high concentration refractory organic matters and antibiotic residues, two laboratory-scale EGSB reactors (the experimental reactor and the control reactor) were constructed for pre-treating OTC production wastewater and the complex characteristics and connections among anaerobic fermentative bacteria, methanogens and fungi were analyzed. The experimental reactor gradually increased OTC doses of 0-200 mg/L by four phases compared with the control reactor which was fed without OTC addition during 280 days' operation. The average COD removal efficiency of 91.44% with the average OTC removal efficiency of 27.90% was achieved at OTC concentration of 200 mg/L. The addition of OTC did not affect the preponderant methanogen type, and Methanosaeta, a strict aceticlastic methanogen genus, was dominant both in working and controlling reactors on day 280. Redundancy analysis revealed that OTC and VFAs were the main environmental factors affecting the microbial communities and molecular ecological networks analysis indicated that the key genera principally belonged to Methanosaeta, Proteobacteria and Apiotrichum. Additionally, the fungi genus Apiotrichum might be related to the degradation of complex organic contaminants in OTC production wastewater treatment system.

Impact of salinity on anaerobic microbial community structure in high organic loading purified terephthalic acid wastewater treatment system.

To investigate the effect of salinity (1% sodium chloride) on anaerobic microbial community structure in high strength telephthalic wastewater treatment system, the performances of anaerobic-aerobic process and the shifts of microbial community in anaerobic tank were studied and determined. Results showed that the chemical oxygen demand (COD) removal in the whole process remained above 90%. And the effluent concentrations of targeted pollutants were lower than 10 mg/L, other than para-toluic acid (PT, 38.09 mg/L). However, methane production significantly decreased compared to no salinity situation. This might be due to the inhibition of salinity on methanogens, which hindered the conversion of acetate to methane. Furthermore, the dominant genus in bacterial level changed from Tepidisphaera to Syntrophus, which facilitated the syntrophic association with hydrogenotrophic methanogens. The prevailed archaea remained acetoclastic Methanothrix above 90%. Therefore, the salinity on anaerobic microbial community structure mainly reflects in the methanogen process, remarkably decreasing methane production.

Hydrolysis and decomposition of waste activated sludge with combined lysozyme and rhamnolipid treatment: Effect of pH.

Effect of pH on waste activated sludge (WAS) hydrolysis and decomposition treating with lysozyme and rhamnolipid combined (Ly + RL) was investigated in this study. Results showed that Ly + RL system could significantly improve the release of soluble organic matters at the optimal RL dosage of 0.3 g/gSS and lysozyme dosage of 0.15 g/gSS. Alkali conditions showed better effect than that of acid on the release of soluble organics, improvement of WAS biodegradability and reduction of big floc size within Ly + RL treatment system and the optimal pH was 10. And 9591.6 mg/L soluble chemical oxygen demand (SCOD), 1612.0 mg/L protein and 1211.6 mg/L polysaccharide were released at pH10 after 12 h co-digestion. 83.7% bacteria and 92.2% archaea were decomposed at pH10. Class Gammaproteobacteria (82.4%) was the predominant bacteria after treated by Ly + RL system, and the treated WAS was beneficial for the subsequent organics bio-degradation and volatile fatty acids accumulation.

Enhancement of excess sludge hydrolysis and decomposition by combined lysozyme and rhamnolipid pretreatment.

Feasibility of combined lysozyme and rhamolipid (RL) pretreatment on the enhancement of excess sludge (ES) hydrolysis and decomposition was assessed in this study. Results showed lysozyme and RL combined treatment could significantly promote ES hydrolysis and decomposition, an additional 1196.9 mg/L soluble chemical oxygen demand (SCOD), 792.5 mg/L protein and 133.5 mg/L polysaccharide were released compared with the sum of sole RL and sole lysozyme treatment at the optimal RL dosage of 0.3 g/gSS and lysozyme dosage of 0.15 g/gSS after 8 h co-digestion. 45.3% bacteria and 84.5% archaea decomposition degree were gained under the combined treatment at the optimal RL dosage. Class Gammaproteobacteria and genus Methanothrix were the predominant bacteria and archaea with the relative abundance of 72.4% and 60.8%, respectively. After the combined pretreatment, ES was beneficial for volatile fatty acids accumulation and acetic acid dependent methane generating inferred from the results of microbial community composition.

Enhancement of excess sludge hydrolysis and decomposition with different lysozyme dosage.

The performance of the lysozyme catalysis on excess sludge (ES) hydrolysis and decomposition was investigated in this study. For this purpose, the release of soluble organic matters from sludge flocs, extracellular polymeric substances (EPS) changes in composition and distribution and the quantity variations of microorganisms were monitored. Results indicated that lysozyme boosted the ES hydrolysis significantly with approximately 236.5 mg/L soluble chemical oxygen demand (SCOD), 58.6 mg/L polysaccharide and 662.7 mg/L protein release within 240 min at the lysozyme dosage of 150 mg/gSS. Arising lysozyme dosages (from 0 to 150 mg/gSS step by step) could dramatically enhance the efficiency of the enzyme on ES with the concentration of polysaccharide increased from 84.6 mg/L to 143.2 mg/L and protein increased from 325.0 mg/L to 987.7 mg/L in total EPS. The decomposition effect of lysozyme on microorganisms improved with dosage, about 15.4%, 17.5% and 20.2% bacteria and 56.3%, 57.2% and 65.0% archaea were disintegrated at the lysozyme dosages of 50, 100 and 150 mg/gSS, respectively. However, fungi were barely influenced by the enzymatic catalysis. Tryptophan-protein like substances and aromatic protein were the dominant ES lysis compositions in EPS.

The relationship between volatile fatty acids accumulation and microbial community succession triggered by excess sludge alkaline fermentation.

The volatile fatty acids (VFAs) accumulation pattern and microbial community succession were studied during excess sludge (ES) alkaline fermentation at pH of 10.0 with expanded granular sludge blanket reactor over 5 cyclers. Microbial community shifted conspicuously as ES suffered alkaline fermentation. Both VFAs and acid-producing bacteria increased rapidly during the first 8 days fermentation time, and they showed a quite positive correlation relationship. In addition, soluble chemical oxygen demand (SCOD) also dramatically increased during the first 8 days, which implied 8 day was the optimum sludge retention time (SRT) for ES alkaline fermentation and VFAs accumulation time. Illumina Miseq Sequencing analysis indicated that Clostridium, Bacillus, Amphibacillus and Peptostreptococcaceae were the dominant bacteria genus to produce VFAs. Acetic acid took about 84% in total VFAs because among the total acid-producing bacteria most bacteria could produce acetic acid.

Amoxicillin effects on functional microbial community and spread of antibiotic resistance genes in amoxicillin manufacture wastewater treatment system.

This study aimed to reveal how amoxicillin (AMX) affected the microbial community and the spread mechanism of antibiotic resistance genes (ARGs) in the AMX manufacture wastewater treatment system. For this purpose, a 1.47 L expanded granular sludge bed (EGSB) reactor was designed and run for 241days treating artificial AMX manufacture wastewater. 454 pyrosequencing was applied to analyze functional microorganisms in the system. The antibiotic genes OXA-1, OXA-2, OXA-10, TEM-1, CTX-M-1, class I integrons (intI1) and 16S rRNA genes were also examined in sludge samples. The results showed that the genera Ignavibacterium, Phocoenobacter, Spirochaeta, Aminobacterium and Cloacibacillus contributed to the degradation of different organic compounds (such as various sugars and amines). And the relative quantification of each ß-lactam resistance gene in the study was changed with the increasing of AMX concentration. Furthermore the vertical gene transfer was the main driver for the spread of ARGs rather than horizontal transfer pathways in the system.

Establishment of thermophilic anaerobic terephthalic acid degradation system through one-step temperature increase startup strategy - Revealed by Illumina Miseq Sequencing.

Over recent years, thermophilic digestion was constantly focused owing to its various advantage over mesophilic digestion. Notably, the startup approach of thermophilic digester needs to be seriously considered as unsuitable startup ways may result in system inefficiency. In this study, one-step temperature increase startup strategy from 37 °C to 55 °C was applied to establish a thermophilic anaerobic system treating terephthalic acid (TA) contained wastewater, meanwhile, the archaeal and bacterial community compositions at steady periods of 37 °C and 55 °C during the experimental process was also compared using Illumina Miseq Sequencing. The process operation demonstrated that the thermophilic TA degradation system was successfully established at 55 °C with over 95% COD reduction. For archaea community, the elevation of operational temperature from 37 °C to 55 °C accordingly increase the enrichment of hydrogenotrophic methanogens but decrease the abundance of the acetotrophic ones. While for bacterial community, the taxonomic analysis suggested that Syntrophorhabdus (27.40%) was the dominant genus promoting the efficient TA degradation under mesophilic condition, whereas OPB95 (24.99%) and TA06 (14.01%) related populations were largely observed and probably take some crucial role in TA degradation under thermophilic condition.

[Isolation and purification of recombinant human lactoferrin (rhLF) from transgenic rice and its antibacterial activities].

OBJECTIVE: To study the isolation and purification of recombinant human lactoferrin (rhLF) from transgenic rice, and to check its antibacterial activities. METHODS: After isolated rhLF from transgenic rice via saturated ammonium sulfate precipitation, then purified it through CM Sepharose FF-exchange chromatography and molecular sieve chromatography Sephadex G25. The inhibition effects under different concentrations of rhLF (0.25, 0.5, 1.0, 2.0, 4.0, and 5.0 mg/ml) against Salmonella typhimurium, Staphyloccocus aureus, Bacillus cereus, Listeria monocytogenes were observed, using broth microdilution method. RESULTS: The rhLF was obtained at a higher purity (about 90%) through successful isolation and purification. After Coomassie blue staining, Westernblot and mass spectrometer analysis, it was identified as the purpose protein with the molecular weight of approximately 79 kDa. The antibacterial experiments showed that 5 mg/ml and 4 mg/ml rhLF could inhibite Salmonella typhimurium and Staphylococcus aureus persistently, 2 mg/ml and 1 mg/ml rhLF showed a significant inhibitory effects in the later period; while 0.5 mg/ml or lower concentration, showed no inhibitory effects. As to Bacillus cereus, only 5 mg/ml and 4 mg/ml rhLF exhibited certain inhibitory effects within 18 hours. Listeria monocytogenes was inhibited within 18 hours just at 5 mg/ml rhLF. CONCLUSION: The rhLF could be successfully separated and purified from transgenic rice, and the purified protein still has significant antibacterial activities.