Identification of surfactants emerged in aerobic granulation

In this study, aerobic granules were cultivated in sequencing batch reactors with activated sludge as the seed. The reactors were operated for 12 h per cycle with the organic loading rate (OLR) increasing in double stepwise from 0.5 to 4.0 g COD L⁻¹ d⁻¹. Within the 40 d running, black granules with regular and smooth morphology were cultivated, which had high wet density and high settling velocity. During the granulation process, foams emerged and disappeared in the reactor, coinciding with the proliferation of filamentous microorganisms in the granules, implying that surfactants might exist and play an important role in the granulation. Using ultra performance liquid chromatography-tandem mass spectrometry, the surfactants were identified as homologous compounds of polyethylene glycol (PEG) with molecular weights ranging mainly from 100 to 500 Da. Their general formulas were proposed as HO-[CH₂-CH₂-O]_n-H. The source of PEG still needs further investigation.     

The role of SBR mixed liquor volume exchange ratio in aerobic granulation

Four sequencing batch reactors (SBR) were operated at different volume exchange ratios of 20-80%. Results showed that a rapid aerobic granulation could be achieved in SBR run at high volume exchange ratios, and the characteristics of mature aerobic granules in terms of mean size, SVI, granule fraction, EPS and calcium content were all closely related to the volume exchange ratio applied to SBR. To interpret the role of volume exchange ratio in aerobic granulation, a concept of minimum settling velocity of bioparticles was proposed, which is the function of both settling time and volume exchange ratio. It was further demonstrated that the effect of volume exchange ratio on aerobic granulation in SBR would be realized through selecting the minimum settling velocity of bioparticles. In fact, the minimum settling velocity would serve as a major selection pressure exerted on bioparticles. It appeared from this study that the production of EPS was stimulated significantly by high volume exchange ratio, which was associated with marketable accumulation of calcium ion in aerobic granulation. It is expected that this study may offer insights into the mechanism of aerobic granulation.     

Detection of DDT and its metabolites in two estuaries of South China using a SPME-based device: First report of p,p′-DDMU in water column

A solid-phase microextration-based sampling method was employed to determine the concentrations of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) and its metabolites, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD), 1,1-dichloro-2,2-bis(p-chlorophenyl)ethene (DDE) and 1-chloro-2,2-bis(p-chlorophenyl)ethene (DDMU), in two estuarine bays, Daya Bay and Hailing Bay, of South China. Six DDT components including p,p′-DDT, o,p′-DDD, p,p′-DDD, o,p′-DDE, p,p′-DDE, and p,p′-DDMU were detected in Hailing Bay, while only p,p′-DDD was found in Daya Bay. p,p′-DDD was the most abundant DDT component in both bays, sharply different from the previous finding in the water column of the Palos Verdes Shelf, California, USA that p,p′-DDE was prevalent. In addition, the occurrence of p,p′-DDMU (with a range of 0.047-0.21 ng/L in Hailing Bay) has not been reported around the globe, and its presence in our study region appeared to stem from dehydrochlorination of p,p′-DDD, favored under aerobic conditions, but further investigations are clearly needed to confirm the mechanism for generation of DDMU in estuarine environments.     

The influence of storage on the morphology and physiology of phthalic acid-degrading aerobic granules

The precultured aerobic granules with special degradabilities could be used as a feasible bioseed for enhancement of aerobic granulation systems. In practice, the storage stability, physicochemical characteristics, and recovering efficiency of granules are crucial for a long-distance transportation and successful application. In this study, phthalic acid (PA)-degrading aerobic granules were cultivated and stored for 8 wk at 4 °C. The granular size, settling ability as well as structure integrity was found stable during the storage period. It was observed that the upper 1/3 part of granules stored in the reagent bottle turned to black color, while the lower 2/3 part granules did not significantly change color (brown–yellow) after the 8-wk storage. The black and brown–yellow color PA-degrading granules were manually separated and re-inoculated into two identical sequencing batch reactors for reviving the PA degradation capability. After a 7 d operation, both black and yellow granules restored their activities to the levels before storage, in terms of total organic carbon removal efficiency (100%), specific oxygen uptake rate (59 mg g VSS⁻¹ h⁻¹), and adenosine triphosphate content (0.016 mg g VSS⁻¹). This study demonstrated that aerobic granules grown on a complex substrate could tolerate storage conditions and rapidly restored their bioactivities toward the target pollutant. The results also shed the light on the future application of precultured aerobic granules with unique functions for biodegradation and bioremediation purpose.     

The effects of different aeration patterns on aerobic granulation

In this study, the effects of aeration patterns in terms of intra- (S1) and extra- (S2) aeration on aerobic granulation were investigated. Granules developed in S1 were retarded due to the high shear force of highly mixed air bubbles, and they were more solid while the S2 granules were porous with many distinct pores and channels. Mature granules in two series had comparable specific oxygen uptake rate and ammonium-nitrogen uptake rate, which were 37.57 and 39.64 mg O₂/(gMLSS ⋅ h), 6.44 and 4.03 mg NH₃-N/ (gMLSS ⋅ h) respectively. Moreover, S1 granules contained more extracellular polymeric substances and higher exopolysaccharide content, which resulted higher negative charges.     

Understanding the granulation process of activated sludge in a biological phosphorus removal sequencing batch reactor

The granulation of activated sludge was investigated using two parallel sequencing batch reactors (SBRs) operated in biological nitrogen and phosphorus removal conditions though the reactor configuration and operating parameters did not favor the granulation. Granules were not observed when the SBR was operated in biological nitrogen removal period for 30 d. However, aerobic granules were formed naturally without the increase of aeration intensity when enhanced biological phosphorus removal (EBPR) was achieved. It can be detected that plenty of positive charged particles were formed with the release of phosphorus during the anaerobic period of EBPR. The size of the particles was about 5-20 μm and their highest positive ζ potential was about 73 mV. These positive charged particles can stimulate the granulation. Based on the experimental results, a hypothesis was proposed to interpret the granulation process of activated sludge in the EBPR process in SBR. Dense and compact subgranules were formed stimulated by the positive charged particles. The subgranules grew gradually by collision, adhesion and attached growth of bacteria. Finally, the extrusion and shear of hydrodynamic shear force would help the maturation of granules. Aerobic granular SBR showed excellent biological phosphorus removal ability. The average phosphorus removal efficiency was over 95% and the phosphorus in the effluent was below 0.50 mg L⁻¹ during the operation.     

Persistence of organochlorine chemical residues in fish from the Tombigbee River (Alabama, USA): Continuing risk to wildlife from a former DDT manufacturing facility

Organochlorine pesticide and total polychlorinated biphenyl (PCB) concentrations were measured in largemouth bass from the Tombigbee River near a former DDT manufacturing facility at McIntosh, Alabama. Evaluation of mean p,p′- and o,p′-DDT isomer concentrations and o,p′- versus p,p′-isomer proportions in McIntosh bass indicated that DDT is moving off site from the facility and into the Tombigbee River. Concentrations of p,p′-DDT isomers in McIntosh bass remained unchanged from 1974 to 2004 and were four times greater than contemporary concentrations from a national program. Total DDT in McIntosh bass exceeded dietary effect concentrations developed for bald eagle and osprey. Hexachlorobenzene, PCBs, and toxaphene concentrations in bass from McIntosh also exceeded thresholds to protect fish and piscivorous wildlife. Whereas concentrations of DDT and most other organochlorine chemicals in fish have generally declined in the U.S. since their ban, concentrations of DDT in fish from McIntosh remain elevated and represent a threat to wildlife.     

Effects of extracellular polymeric substances on aerobic granulation in sequencing batch reactors

The effects of extracellular polymeric substances on aerobic granulation in sequencing batch reactors were investigated by evaluating the content and compositions of extracellular polymeric substances, and the relationship between extracellular polymeric substances composition and surface properties of glucose-fed aerobic granules. The results showed that extracellular polymeric substances could affect surface properties of cells, such as surface charge and hydrophobicity, enhance polymeric interaction and promote aerobic granulation. Moreover, extracellular polymeric substances were produced mainly in the exponential phase, and served as carbon and energy source in starvation phase during granulation process, thus regulating the growth of bacteria in the interior and exterior of granules, and maintaining the integrality of granules.     

Inorganic nanoparticles enhance the production of reactive oxygen species (ROS) during the autoxidation of L-3,4-dihydroxyphenylalanine (L-dopa)

Public concerns over the toxicity of nanoparticles (NPs) are growing due to the rapid development of nanotechnology. An important mechanism of nanotoxicity is oxidative stress resulting from reactive oxygen species (ROS). In this study, the chemical production of ROS by inorganic NPs oxidizing the mammalian phenolic compound, l-3,4-dihydroxyphenylalanine (l-dopa) was evaluated using a ROS sensitive dye, 2′,7′-diclorodihydrofluorescin (DCFH). CeO₂, Fe₂O₃ and Fe⁰ NPs enhanced ROS production during the autoxidation of l-dopa by more than four-fold in reactions that were dependent on O₂. This is the first report of chemical ROS production due to interaction of phenolic compounds with NPs. Mn₂O₃ oxidized DCFH in a reaction that did not require O₂ or l-dopa, suggesting a direct redox reaction between the Mn₂O₃ and the dye. CeO₂, Mn₂O₃ and to a lesser extent Fe⁰ formed clear electron paramagnetic resonance (EPR) signature for hydroxyl radicals when incubated in aerobic aqueous suspensions with spin traps. The results indicate that NPs can generate ROS via chemical reactions with medium components and biomolecules susceptible to oxidation, such as l-dopa. NPs were reactive whereas micron-sized particles were not. The combined assay with l-dopa and DCFH is a method proposed to screen for chemical ROS production by NPs.     

Effect of ultrasonication and Fenton oxidation on biodegradation of bis(2-ethylhexyl) phthalate (DEHP) in wastewater sludge

The presence of bis(2-ethylhexyl) phthalate (DEHP) and its metabolites, i.e. 2-ethylhexanol, 2-ethylhexanal, and 2-ethylhexanoic acid in wastewater sludge (WWS) were investigated during aerobic digestion and Bacillus thuringiensis (Bt)-based fermentation of WWS. Ultrasonication and Fenton oxidation pre-treatment was applied to improve biodegradability of WWS and bioavailability of the target compounds for digestion and fermentation. DEHP and 2-ethylhexanoic acid were observed at higher concentration, meanwhile 2-ethylhexanol and 2-ethylhexanal were observed at lower concentration in WWS. After 20-day aerobic digestion, DEHP removal was 72%, 89%, and 85%, and 2-ethylhexanoic acid removal was 71%, 84%, 79%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges. Bt was found to degrade DEHP, leading to DEHP removal of 21%, 40%, and 30%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges in the fermentation. The results suggested that aerobic stabilization and Bt-based fermentation can remove the phthalates, and pre-treatment of WWS was also effective in improvement of DEHP biodegradation. Hence, Bt-based biopesticide production from WWS can be applied safely when taking into consideration the phthalate contaminants.     

Cytogenetic status of human lymphocytes after exposure to low concentrations of p,p'-DDT, and its metabolites (p,p'-DDE, and p,p'-DDD) in vitro

Despite that the use of DDT has been restricted for more than 40 years to malaria affected areas, low doses of this pesticide and its metabolites DDE and DDD can be found in the environment around the world. Although it has been shown that these pollutants induce cell and DNA damage, the mechanisms of their cytogenotoxic activity remains largely unknown. This study looks into their possible genotoxic effects, at doses that can be found in body fluids, on human lymphocytes using the cytokinesis-block micronucleus assay and the comet assay. After exposure for 1, 6, and 24 h compounds p,p′-DDT (0.1 μg mL⁻¹), p,p′-DDE (4.1 μg mL⁻¹), and p,p′-DDD (3.9 μg mL⁻¹) showed increase in DNA damage. The most significant results were observed at exposure period of 24 h where number of micronucleated cells increased from control 2.5 ± 0.71 to 23.5 ± 3.54, 13.5 ± 0.71, and 16.5 ± 6.36 for DDT, DDE, and DDD, respectively. Similar effect was observed using comet test where the percentage of DNA in comets tail increased from control 1.81 ± 0.16 to 17.24 ± 0.55, 11.21 ± 0.56 and 9.28 ± 0.50 for each compound, respectively. At the same time Fpg-comet assay failed to report induction of oxidative DNA damage of these pollutants. Additionally, the type of cell death was determined using diffusion assay and necrosis dominated. Our findings suggest that even at low concentrations, these pesticides could induce cytogenetic damage to human peripheral blood lymphocytes and in that manner have the impact on human health as well.     

厌氧序批式反应器中丝状颗粒污泥的形成

以葡萄糖为基质,研究浮动盖式ASBR中颗粒污泥的形成。实验结果表明,污泥在120 d时完全颗粒化,颗粒污泥具有良好的沉降性能(25 m/h)及良好的产甲烷活性,甲酸、乙酸、丙酸及丁酸最大代谢活性分别达到0.216、0.825、0.237和0.796 g COD/(g VSS.d)。电镜扫描(SEM)发现,整个污泥结构均一,均由丝状菌构成。     

Characterization of bacterial communities at heavy-metal-contaminated sites

The microbial community in soil samples from two long-term contaminated sites was characterized by using culture-dependent and culture-independent methods. The two sites investigated contained high amounts of heavy metals and were located in the upper Silesia Industrial Region in southern Poland. The evaluation of the aerobic soil microbial population clearly demonstrated the presence of considerable numbers of viable, culturable bacteria at both sites. A high fraction of the bacterial population was able to grow in the presence of high amounts of metals, i.e. up to 10 mM Zn²⁺, 3 mM Pb²⁺ or 1 mM Cu²⁺. Site 1 contained significantly (P < 0.05) lower bacterial numbers growing in the presence of 10 mM Zn²⁺ than site 2, while the opposite was observed for bacteria tolerating 1 mM Cu²⁺. This coincided with the contents of these two metals at the two sites. Ecophysiological (EP) indices for copiotrophs (r-strategists) and oligotrophs (K-strategists) pointed to high bacterial diversity at both sites. Fluorescence in situ hybridization (FISH) analysis indicated that Actinobacteria and Proteobacteria represent the physiologically active fraction of bacteria at the two sites. Shannon diversity (H′) indices for FISH-detected bacterial phylogenetic groups were not significantly different at the two sites.     

New DDT inputs after 30 years of prohibition in Spain. A case study in agricultural soils from south-western Spain

This study provides information on the current status of contamination by DDT in agricultural soils in south-western Spain. A recent use of technical DDT in at least 17% of the soils was found based on the values (<1) of the ratio Rp,p^′/p,p^′=[p,p^′-DDE+p,p^′-DDD]/[p,p^′-DDT]. According to the ratio Ro,p^′/p,p^′=[o,p^′-DDT]/[p,p^′-DDT], a dicofol type contamination was detected in about 27% of the soils. A wide range of concentrations was observed (0.08-11.1 ng/g d.w.) regardless of the type of crop soil. Enantiomeric fractions (EFs), based on the chiral analysis of o,p′-DDT residues differed from the racemic value (0.500) in most soils but they were not correlated with the study variables [DDTs], SOM, Rp,p^′/p,p^′ and Ro,p^′/p,p^′. Given the health risks posed by DDT, our findings support how the environmental control of legacy pollutants such as DDT cannot be neglected.     

Effect of coplanar PCB concentration on dechlorinating microbial communities and dechlorination in estuarine sediments

The effect of concentration of coplanar PCB on the dechlorinating microbial community and dechlorination were investigated in anoxic estuarine sediment collected from Er-Jen River and enriched with 10 and 50 mg L⁻¹ of 3,4,4′,5-tetrachlorobiphenyl, 3,3′,4,4′,5-pentachlorobiphenyl, and 3,3′,4,4′,5,5′-hexachlorobipheny. Dechlorination rates were similar in the cultures enriched with 10 and 50 mg L⁻¹ of 3,4,4′,5-tetrachlorobiphenyl, whereas significantly higher dechlorination rates were observed in cultures enriched with 10 mg L⁻¹ of 3,3′,4,4′,5-pentachlorobiphenyl. No dechlorination was observed in sediment slurries enriched with 3,3′,4,4′,5,5′-hexachlorobipheny. Para dechlorination occurred prior to meta dechlorination during reductive dechlorination of 3,4,4′,5-tetrachlorobiphenyl and 3,3′,4,4′,5-pentachlorobiphenyl. GC-MS and denaturing gradient gel electrophoresis (DGGE) were used to detect dechlorination products and dechlorinating microorganisms in the enriched sediment cultures during the process of degradation. Two Chloroflexi phylotypes observed in DGGE were responsible for para and meta dechlorination respectively. Phylotype Cp-1 has 98% similarity to uncultured bacterium N5-12. Phylotype Cm-1 has 99% similarity to uncultured dechlorinating bacterium m1 or SF1 belonging to the ο-17/DF-1 group of PCB-dechlorinating bacteria.     

Enhanced sludge granulation in upflow anaerobic sludge blanket (UASB) reactors by aluminum chloride.

Two upflow anaerobic sludge blanket (UASB) reactors were concurrently operated for 146 days to examine the effects of aluminum chloride (AlCl3) on the sludge granulation process during start-up. Sludge granulation (defined as that over 10% of granules were larger than 2.0 mm) was achieved in the control reactor (R1) in approximate three months. Introduction of Al3+ at a concentration of 300 mg/l reduced the sludge granulation time by approximate one month. Throughout the experiment the AlCl3-added reactor (R2) had a higher biomass concentration, e.g., 13.8 g-MLVSS/l versus 12.8 g-MLVSS/l on Day 146. Granules became visible earlier in R2 compared with R1 (35 days versus 65 days). The average size of granules from R2 was larger than that from R1. The results demonstrated that AlCl3 enhanced the sludge granulation process in the UASB reactors.