Influences of extracellular polymeric substances (EPS) recovered from waste sludge on the ability of Jiaozhou Bay to self-remediate of diesel-polluted seawater.

The introduction of EPS recovered from waste sludge may have an impact on the process of microbial remediation of oil-contaminated seawater. This study investigated the effect of EPS on the self-remediation capacity of diesel-polluted seawater in Jiaozhou Bay. Hydrocarbon attenuation and microbial activity were monitored in seawater collected from five islands after diesel and N, P addition, with and without EPS, incubated under aerobic conditions. Compared to seawater without EPS, degradation of TPH (total petroleum hydrocarbon) doubled and improved degradation of non-volatile (C16-C24) hydrocarbons to some extent in EPS-added seawater. The introduction of EPS led to changes in microbiota richness and diversity, significantly stimulating the growth of Proteobacteria and Firmicutes phyla or Bacillus and Pseudomonas genera. RT-qPCR analysis indicated EPS caused higher increases in cytochrome P450 gene copies than alkB. Prediction of alkane decay genes from 16S rRNA sequencing data revealed that EPS addition obviously promoted genes related to ethanol dehydrogenation function in the microbial community. Additionally, EPS enhanced the enzymatic activities of alkane hydroxylase, ethanol dehydrogenase, phosphatase and lipase, but increased protease and catalase inconspicuously. The above outlook that environmental sustainability of EPS from waste sludge for diesel-contaminated seawater remediation may provide new perspectives for oil spill bioremediation.

Glutamicibacter nicotianae AT6: A new strain for the efficient biodegradation of tilmicosin.

The degradation of tilmicosin (TLM), a semi-synthetic 16-membered macrolide antibiotic, has been receiving increasing attention. Conventionally, there are three tilmicosin degradation methods, and among them microbial degradation is considered the best due to its high efficiency, eco-friendliness, and low cost. Coincidently, we found a new strain, Glutamicibacter nicotianae sp. AT6, capable of degrading high-concentration TLM at 100 mg/L with a 97% removal efficiency. The role of tryptone was as well investigated, and the results revealed that the loading of tryptone had a significant influence on TLM removals. The toxicity assessment indicated that strain AT6 could efficiently convert TLM into less-toxic substances. Based on the identified intermediates, the degradation of TLM by AT6 processing through two distinct pathways was then proposed.

Difference in calcium ion precipitation between free and immobilized Halovibrio mesolongii HMY2.

Biomineralization has become a research focus in wastewater treatment due to its much lower costs compared to traditional methods. However, the low sodium chloride (NaCl)-tolerance of bacteria limits applications to only water with low NaCl concentrations. Here, calcium ions in hypersaline wastewater (10% NaCl) were precipitated by free and immobilized Halovibrio mesolongii HMY2 bacteria and the differences between them were determined. The results show that calcium ions can be transformed into several types of calcium carbonate with a range of morphologies, abundant organic functional groups (C-H, C-O-C, C=O, etc), protein secondary structures (ß-sheet, α-helix, 310 helix, and ß-turn), P=O and S-H indicated by P2p and S2p, and more negative δ13CPDB (‰) values (-16.8‰ to -18.4‰). The optimal conditions for the immobilized bacteria were determined by doing experiments with six factors and five levels and using response surface method. Under the action of two groups of immobilized bacteria prepared under the optimal conditions, by the 10th day, Ca2+ ion precipitation ratios had increased to 79%-89% and 80%-88% with changes in magnesium ion cencentrations. Magnesium ions can significantly inhibit the calcium ion precipitation, and this inhibitory effect can be decreased under the action of immobilized bacteria. Minerals induced by immobilized bacteria always aggregated together, had higher contents of Mg, P, and S, lower stable carbon isotope values and less well-developed protein secondary structures. This study demonstrates an economic and eco-friendly method for recycling calcium ions in hypersaline wastewater, providing an easy step in the process of desalination.

Simultaneous nitrification and denitrification in an aerobic biofilm biosystem with loofah sponges as carriers for biodegrading hydrolyzed polyacrylamide-containing wastewater.

Simultaneous nitrification and denitrification (SND) during treating hydrolyzed polyacrylamide (HPAM) containing wastewater were explored in an aerobic biofilm reactor biosystem. Here, loofah sponges as the environment-friendly and low-cost material were applied as the carriers in this biosystem. The removal efficiencies of HPAM and total nitrogen (TN) reached 43.6% and 54.3%, respectively, after 120 days stabilized running periods. Moreover, the structure of loofah sponges affected anaerobic microenvironment significantly which was indispensable for realizing a high-performance of SND. Key microorganisms in this biosystem included nitrobacteria, denitrobacteria and HPAM-biodegrading bacteria. The abundance of nitrobacteria and denitrobacteria on the biofilm was increased by 17.2% and 15.3%, respectively, through cultivation. Meanwhile, the biotransformation mechanisms of HPAM and diverse valence of nitrogen under different chemical oxygen demand (COD)/N and dissolved oxygen (DO) conditions were investigated. When COD/N and DO were 8:1 and 2 mg/L, HPAM biodegradation, SND efficiency and TN removal achieved their maximum, and the values were 54.3%, 92.3% and 60.1%, respectively. Key enzyme activities also reached their maximum in this condition. The optimal COD/N and DO was pivotal to achieve the high-performance of SND, and it was closely correlated with HPAM biodegradation. Meanwhile, SND could facilitate the biotransformation of HPAM.

Hydrolyzed polyacrylamide biotransformation in an up-flow anaerobic sludge blanket reactor system: key enzymes, functional microorganisms, and biodegradation mechanisms.

Hydrolyzed polyacrylamide (HPAM) biotransformation in an up-flow anaerobic sludge blanket reactor including biodegradation performances, biodegradation mechanisms, key enzymes, and functional microorganisms was explored. Response surface methodology was applied to further improve HPAM degradation. The predicted degradation ratios of HPAM and CODCr were 46.2% and 83.4% under the optimal conditions. HPAM biodegradation ratio and total organic carbon removal ratio reached 40.5% and 38.9%. Total nitrogen concentration was dramatically decreased with the increasing fermentation time during the fermentation, while low ammonia nitrogen (NH4+-N) and nitrite nitrogen (NO2--N) were generated. NH4+-N and NO2--N increased slightly on the whole. Enzyme activity change was correlated with HPAM biodegradation. Dehydrogenase activity had a decline of 21.3-41.0%, and the minimum value occurred at 300 mg/L of HPAM. Urease activity was varied from 28.7 to 78.7% and the maximal inhibition ratio occurred at 200 mg/L of HPAM. Mechanisms for the biodegradation of HPAM were also explored by FT-IR, HPLC, and SEM. The results indicated that long-chain HPAM was broken into micromolecule compounds and the amide groups of HPAM were transformed into carboxyl groups. Based on the sequencing results on an Illumina MiSeq platform, Proteobacterias, Bacteroidetes, and Chloroflexi were turned out to be the critical microorganisms involved in HPAM degradation. This work lays a basis for HPAM-containing wastewater treatment and offers a support for water saving and emission reduction. It is of great significance to the sustainable development of oilfield.

Regulation of different electron acceptors on petroleum hydrocarbon biotransformation to final products in activated sludge biosystems.

The types and concentrations of electron acceptor are the significant factors influencing the oxidation and biotransformation of organic matter in the process of pollutant biodegradation. Regulation of O2, SO42- and NO3- as electron acceptors on petroleum hydrocarbon biotransformation to final products was studied using the multiple methods including mesoscale biodegradation experiments, thermodynamic theoretical calculations and stoichiometric analyses. Petroleum hydrocarbon biodegradation ratio (PHBR) rose from 64.7 to 82.4% with dissolved oxygen (DO) (3-5 mg L- 1). PHBR increased from 57.4 to 66.1% in SO42--reducing biosystems and rose from 65.0 to 77.9% in NO3--reducing biosystems. Carbon balance was verified in different cultures. The shared functional microorganisms in different biosystems included Candida, Rhodococcus, Pseudomonas, Ochrobactrum, Marinobacter, Bacillus, Azoarcus, Alcanivorax, Acinetobacter. Pandoraea, Enterobacter and Burkholderia in anaerobic biosystems preferred to use NO3- and SO42- as electron acceptors for metabolism, and order of availability followed: NO3- > SO42-. Thermodynamic constraint showed that potentials of alkanes biotransformation to methane through hydrogenotrophic and acetoclastic methanogenesis in NO3--reducing biosystems were 7.27-7.73 and 7.25-7.70 times larger than those of SO42--reducing biosystems, respectively. Metabolism equations of microorganisms proved that anabolism and catabolism on alkanes were feasible. This work provides a support for studying the biochemical process of petroleum hydrocarbon biotransformation and lays a foundation for the realization of oil-containing wastewater bioremediation.

Calcium ion removal at different sodium chloride concentrations by free and immobilized halophilic bacteria.

Much attention has been paid to Ca2+ ion removal by biomineralization due to the dangers of Ca2+ on industrial processes and human health. However, Ca2+ removal from hypersaline water by biomineralization is quite difficult due to there being few halophilic bacteria tolerating higher salinities. In this study, free and immobilized Virgibacillus massiliensis C halophilic bacteria exhibiting carbonic anhydrase activity were used to remove Ca2+ ions from water at different NaCl concentrations. With increasing NaCl concentrations (10, 50, 100, 150 and 200 g/L), Ca2+ ion concentrations in the presence of free bacteria and in two groups of immobilized bacteria for a period of 6 days sharply decreased from 1200 mg/L to 219-562 mg/L, 71-214 mg/L and 21-159 mg/L, respectively; Ca2+ precipitation ratios were 55%-81%, 82%-94% and 87%-98%, respectively. The humic acid-like substances, protein, DNA and polysaccharide, released by the bacteria, promoted the Ca2+ ion removal. The immobilized bacteria were able to be recycled and precultured, which would save industry costs and increase Ca2+ ion removal efficiency. Biological processes for Ca2+ ion removal include cell surface, intracellular and extracellular biomineralization. The biogenesis of calcium carbonate was proved by SEM-EDS, FTIR, XPS and stable carbon isotope values. This study provides insights into the effective removal of Ca2+ ions by biomineralization in hypersaline water.

Hydrolyzed polyacrylamide biotransformation during the formation of anode biofilm in microbial fuel cell biosystem: Bioelectricity, metabolites and functional microorganisms.

The anode biofilm serves as the core dominating the performance of microbial fuel cell (MFC) biosystem. This research provides new insights into hydrolyzed polyacrylamide (HPAM) biotransformation during the formation of anode biofilm. The current density, coulombic efficiency, voltage, power density, volatile fatty acid (VFA) production and total nitrogen (TN) removal enhanced with the thickening of biofilm (1-6 cm), and the maximums achieved 146 mA·m-2, 47.3%, 8.76 V, 1.28 W·m-2, 184 mg·L-1 and 84.6%, respectively. HPAM concentration descended from 508 mg·L-1 to 83.3 mg·L-1 after 60 days. HPAM was metabolized into VFAs, N2, NO2--N and NO3--N, thereby releasing electrons. Laccase and tyrosine/tryptophan protein induced HPAM metabolism and bioelectricity production. The microbial functions involving HPAM biotransformation and bioelectricity generation were clarified. The alternative resource recovery, techno-economic comparison and development direction of MFC biosystem were discussed to achieve the synchronization of HPAM-containing wastewater treatment and bioelectricity generation based on MFC biosystem.

The proliferation and colonization of functional bacteria on amorphous polyethylene terephthalate: Key role of ultraviolet irradiation and nonionic surfactant polysorbate 80 addition.

Environmental pollution with plastics including polyethylene terephthalate (PET) has become a severe global problem, especially microplastic pollution, which is acknowledged as an emerging global pollutant. Biodegradation as a feasible and promising method has been studied, while colonization as the initiating step of the degradation process has seldom been studied. Here in this paper, we explored for the first time the key role of ultraviolet (UV) irradiation and nonionic surfactant polysorbate 80 (Tween-80, 0.2% V/V) in the proliferation and colonization of three functional bacteria (Pseudomonas putida, Pseudomonas sp. and Paracoccus sp.) on amorphous PET (APET). We found that 25 days of UV irradiation can trigger photolytic degradation process (appear the stretching vibration of associating carboxyl end group and the in-plane bending vibration of -OH) and introduce oxygen-containing functional groups on the surface of APET, even though the hydrophobicity of APET was scarcely changed. With regard to Tween-80, it can be utilized by these bacteria strains as carbon source to promote the proliferation, and it can also improve the cell surface hydrophobicity to stimulate the bacterial colonization during the first ten days of the experiment. When UV-irradiation and Tween-80 were provided together, the former factor can provide the target sites for functional bacteria to colonize, and the later factor can provide more candidates waiting to colonize by stimulating proliferation. As a result, an even better proliferation and colonization result can be achieved through the synergistic effect between the two factors. To some extent, the exposure between potential degrading bacteria and substrates to be degraded can be increased, which will create conditions for degrading. Generally, this research can provide certain theoretical basis and technical guidance for the remediation of plastic-polluted soil and the ocean.

Key role of different levels of dissolved oxygen in hydrolyzed polyacrylamide bioconversion: Focusing on metabolic products, key enzymes and functional microorganisms.

Dissolved oxygen (DO) played a short board effect on nitrogen biotransformation and pollutant metabolism. This study for the first time explored the key role of different levels of DO (covering anaerobic, anoxic and aerobic) on hydrolyzed polyacrylamide (HPAM) bioconversion. HPAM was metabolized to intermediates with different chain length. Volatile fatty acid (VFA) production rose first and then descended with DO concentration (0-2 mg·L-1), and the maximum reached 92.5 mg·L-1 when DO was 0.5 mg·L-1. Total nitrogen (TN) removal increased first and then dropped with DO concentration, and the maximum (61.4%) occurred at 0.5 mg·L-1 DO. NH4+-N dipped from 42.8 to 0 mg·L-1 and NO3--N rose from 0 to 32.8 mg·L-1 with DO concentration. The changes of enzyme activities were consistent with those of VFA production and TN removal, which were related to HPAM metabolism and N bioconversion. Microbial function was correlated to HPAM metabolism, N bioconversion and key enzyme.

Biohydrogen and polyhydroxyalkanoate production from original hydrolyzed polyacrylamide-containing wastewater.

This work aimed to study biohydrogen (H2) and polyhydroxyalkanoate (PHA) production from original hydrolyzed polyacrylamide (HPAM)-containing wastewater. NH4+-N from HPAM hydrolysis was removed efficiently through short-cut nitrification and anoxic ammonia oxidation (anammox). Carbon/Nitrogen (C/N) ratios of effluent reached 51-97, and TOC decreased only 2%-4%, providing potential for subsequent H2 and PHA production. The maximum yields of H2 (0.833 mL·mg-1substrate) and Volatile Fatty Acid (VFA) (465 mg·L-1) occurred at influent C/N ratio of 51. Substrate removal increased linearly with the activities of dehydrogenase and hydrogenase (R2 ≥ 0.990), and H2 yield rose exponentially with enzyme activities (R2 ≥ 0.989). The maximum PHA yield (54.2% VSS) occurred at the 42nd hour and influent C/N ratio of 97. PHA yield was positively correlated with substrate uptake. The change of H2-producing, PHA-accumulating and HPAM-degradating bacteria indicated that those functional microorganisms had synergistic effects on H2 production and substrate uptake, as well as PHA accumulation and substrate uptake.

Insights into the effect of different levels of crude oil on hydrolyzed polyacrylamide biotransformation in aerobic and anoxic biosystems: Bioresource production, enzymatic activity, and microbial function.

The differences of crude oil recovery ratio resulted in different levels of crude oil in actual hydrolyzed polyacrylamide (HPAM)-containing wastewater. The effect of crude oil on HPAM biotransformation was explored from bioresource production, enzymatic activity and microbial function. In aerobic biosystems, the highest polyhydroxyalkanoate (PHA) yield (19.6%-40.2%) and dehydrogenase (DH) activity (4.06-8.32 mg·g-1 VSS) occurred in the 48th hour, and increased with crude oil concentration (0-400 mg·L-1). In anoxic biosystems, the highest PHA yield (24.5%-50.5%) and DH activity (3.24-6.69 mg·g-1 VSS) occurred in the 72nd hour, and increased with crude oil concentration. The higher substrate removal (38.5%-65.7%) occurred in aerobic biosystems, while the higher PHA accumulation occurred in anoxic biosystems. PHA yield, DH activity and HPAM removal were related. Microbial function related to HPAM biodegradation and PHA synthesis was discussed. The main function of Pseudomonas and Bacillus in aerobic biosystems was to degrade HPAM, and in anoxic biosystems was to synthesize PHA.

Potential of hydrolyzed polyacrylamide biodegradation to final products through regulating its own nitrogen transformation in different dissolved oxygen systems.

Potential of hydrolyzed polyacrylamide (HPAM) biodegradation to final products was studied through regulating its own nitrogen transformation. Under the conditions of 2, 3 and 4 mg/L of DO, HPAM removal ratio reached 16.92%, 24.51% and 30.78% and the corresponding removal ratio reached 49.15%, 60.25% and 76.44% after anaerobic biodegradation. NO3--N concentration was 9.43, 14.10 and 17.99 mg/L in aerobic stages and the corresponding concentration was 0.17, 0.07 and 0.008 mg/L after anaerobic biodegradation. Oxygen as electron acceptors stimulated the activities of nitrification bacteria and other functional bacteria, thus further enhanced nitrification and HPAM biodegradation. NO3- (from HPAM oxidation) as electron acceptors stimulated the activities of nitrate-reducing, acetate-producing and methanogenic microorganisms and they could form a synergistic effect on denitrification and methanogenesis. Thermodynamic opportunity window revealed that NOx- could accelerate anaerobic HPAM bioconversion to methane. Aerobic and anaerobic growth-process equations of cells verified that the metabolism on HPAM was feasible.

Effects of different electron acceptors on the methanogenesis of hydrolyzed polyacrylamide biodegradation in anaerobic activated sludge systems.

The type of electron acceptor was a crucial factor in regulating the methanogenic process of anaerobic hydrolyzed polyacrylamide (HPAM) degradation. The combined methods of biodegradation experiments and thermodynamic calculations were applied to explore the effects of different electron acceptors on methanogenic HPAM degradation. Under the conditions of without electron acceptor, SO42-, Fe3+, SO42- and Fe3+ as electron acceptors, HPAM biodegradation ratio reached 31.56%, 41.48%, 49.4% and 61.1%, acetate production reached 0.0532, 28.28, 112.7 and 141.95mg·L-1, CH4 production reached 0.024, 0.3015, 9.446 and 11.78mg·L-1, respectively. The synergistic effect of SO42- and Fe3+ further promoted methanogenic HPAM biotransformation. Archaeal community analysis revealed that Methanobacteriales, Methanomicrobiales and Methanosarcinales were dominant. Thermodynamic opportunity windows of methanogenesis with Fe3+ as electron acceptor are 35 times larger than that with SO42- as electron acceptor. It indicated that acetoclastic methanogenesis was dominant and hydrogenotrophic methanogenesis was inhibited in the methane-producing process of anaerobic HPAM degradation.

Hydrolyzed polyacrylamide biodegradation and mechanism in sequencing batch biofilm reactor.

An investigation was performed to study the performance of a sequencing batch biofilm reactor (SBBR) to treat hydrolyzed polyacrylamides (HPAMs) and to determine the mechanisms of HPAM biodegradation. The mechanisms for the optimized parameters that significantly improved the degradation efficiency of the HPAMs were investigated by a synergistic effect of the co-metabolism in the sludge and the enzyme activities. The HPAM and TOC removal ratio reached 54.69% and 70.14%. A significant decrease in the total nitrogen concentration was measured. The carbon backbone of the HPAMs could be degraded after the separation of the amide group according to the data analysis. The HPLC results indicated that the HPAMs could be converted to polymer fragments without the generation of the acrylamide monomer intermediate. The results from high-throughput sequencing analysis revealed proteobacterias, bacteroidetes and planctomycetes were the key microorganisms involved in the degradation.

Kinetics and thermodynamics of biodegradation of hydrolyzed polyacrylamide under anaerobic and aerobic conditions.

Kinetics and thermodynamics of hydrolyzed polyacrylamide (HPAM) biodegradation in anaerobic and aerobic activated sludge biochemical treatment systems were explored to determine the maximum rate and feasibility of HPAM biodegradation. The optimal nutrient proportions for HPAM biodegradation were determined to be 0.08g·L(-1) C6H12O6, 1.00g·L(-1) NH4Cl, 0.36g·L(-1) NaH2PO4 and 3.00g·L(-1) K2HPO4 using response surface methodology (RSM). Based on the kinetics, the maximum HPAM biodegradation rates were 16.43385mg·L(-1)·d(-1) and 2.463mg·L(-1)·d(-1) in aerobic and anaerobic conditions, respectively. The activation energy (Ea) of the aerobic biodegradation was 48.9897kJ·mol(-1). Entropy changes (ΔS) of biochemical treatment system decreased from 216.21J·K(-1) to 2.39J·K(-1). Thermodynamic windows of opportunity for HPAM biodegradation were drawn. And it demonstrated HPAM was biodegraded into acetic acid and CO2 under laboratory conditions. Growth-process equations for functional bacteria anaerobically grown on polyacrylic acid were constructed and it confirmed electron equivalence between substrate and product.

[Epidemic situation of malaria in Taixing City, 2005-2013].

OBJECTIVE: To analyze the malaria situation and control measures in recent years in Taixing City, so as to provide the evidence for formulating the reasonable countermeasures in the future. METHODS: The information of malaria prevalence, the reported data of blood examinations for fever persons and the epidemiological data were collected and the malaria incidence, population distribution, and the imported cases were analyzed. RESULTS: A total of 37 malaria cases were reported from 2005 to 2013 and the incidences were from 0.0078/10,000 to 0.0669/10,000 with the large ascensional range. There were 34 imported malaria patients (91.89% of the total malaria cases) including 2 patients infected outside Taixing City, 2 outside Jiangsu Province, and 30 outside China. Among the 30 patients overseas infected, there was 1 vivax malaria case, there were 2 ovale malaria cases, and 27 falciparum malaria cases. There were incidences throughout the year and no obvious seasonal characteristics. The cases were mainly distributed from 30 to 49 years old (83.78%). CONCLUSIONS: The malaria cases in Taixing City are mainly overseas imported, and the incidence presents a rising trend. Therefore, the relevant authority should enhance the malaria supervision and management, especially for floating population.

[Investigation on infections of human intestinal parasites in Taixing City].

OBJECTIVE: To understand the current status of infections of intestinal parasites of population in Taixing City, Jiangsu Province. METHODS: The infection rates and densities of human intestinal parasites were investigated according to the methods of the National Investigation Scheme on Human Principal Parasites, and the data of society, economy and disease control were collected and analyzed. RESULTS: Among 2 556 people investigated in five villages, 16 persons were found with intestinal parasites, with an infection rate of 0.63%. The infection rate was higher in residents with a low education level than in others and it was higher in the age group over 50 years than in the group under 50 years. The infection density was mild and the most was the single parasite infection. CONCLUSIONS: The current status of intestinal parasite infections of population in Taixing City has reached the county-level control standard of Jiangsu Province. Therefore, the preventive strategy and measures should be adjusted and the monitoring work should be strengthened.