Aerobic granulation of pure bacterial strain Bacillus thuringiensis

The objective of this study is to cultivate aerobic granules by pure bacterial strain, Bacillus thuringiensis, in a sequencing batch reactor. Stable granules sized 2.0–2.2 mm were formed in the reactor after a five-week cultivation. These granules exhibited excellent settling attributes, and degraded phenol at rates of 1.49 and 1.19 g phenol/(g VSS?d) at 250 and 1500 mg/L of phenol concentration, respectively. Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates, and the outer edge of the granule was away from the core regime in the following stage.     

Nitrification and denitrification in biological activated carbon filter for treating high ammonia source water

Since the ammonia in the effluent of the traditional water purification process could not meet the supply demand, the advanced treatment of a high concentration of NH₄ ⁺-N micro-polluted source water by biological activated carbon filter (BACF) was tested. The filter was operated in the downflow manner and the results showed that the removing rate of NH₄ ⁺-N was related to the influent concentration of NH₄ ⁺-N. Its removing rate could be higher than 95% when influent concentration was under 1.0 mg/L. It could also decrease with the increasing influent concentration when the NH₄ ⁺-N concentration was in the range from 1.5 to 4.9 mg/L and the dissolved oxygen (DO) in the influent was under 10 mg/L, and the minimum removing rate could be 30%. The key factor of restricting nitrification in BACF was the influent DO. When the influent NH₄ ⁺-N concentration was high, the DO in water was almost depleted entirely by the nitrifying and hetetrophic bacteria in the depth of 0.4 m filter and the filter layer was divided into aerobic and anoxic zones. The nitrification and degradation of organic matters existed in the aerobic zone, while the denitrification occurred in the anoxic zone. Due to the limited carbon source, the denitrification could not be carried out properly, which led to the accumulation of the denitrification intermediates such as NO₂ ^?. In addition to the denitrification bacteria, the nitrification and the heterotrophic bacteria existed in the anoxic zone.     

Cultivation of aerobic granular sludge in a conventional,continuous flow,completely mixed activated sludge system

Aerobic granules were formed in a conventional, continuous flow, completely mixed activated sludge system (CMAS). The reactor was inoculated with seed sludge containing few filaments and fed with synthetic municipal wastewater. The settling time of the sludge and the average dissolved oxygen (DO) of the reactor were 2 h and 4.2 mg·L^-1, respectively. The reactor was agitated by a stirrer, with a speed of 250 r·min^-1, to ensure good mixing．The granular sludge had good settleability, and the sludge volume index (SVI) was between 50 and 90 mL·g^-1. The laser particle analyzer showed the diameter of the granules to be between 0.18 and 1.25 mm. A scanning electron microscope (SEM) investigation revealed the predominance of sphere-like and rod-like bacteria, and only few filaments grew in the granules. The microbial community structure of the granules was also analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Sequencing analysis indicated the dominant species were α, β, and γ-Proteobacteria, Bacteroidetes, and Firmicutes. The data from the study suggested that aerobic granules could form, if provided with sufficient number of filaments and high shear force. It was also observed that a high height-to-diameter ratio of the reactor and short settling time were not essential for the formation of aerobic granular sludge.     

New denitrifying bacteria isolated from Red Sea sediments

Sediments of various newly discovered deeps in the Red Sea were analyzed for the occurrence of denitrifying bacteria. The samples were collected in 1972 during the Valdivia cruises. Among the 27 different samples investigated, 17 revealed both coccoid and rod-shaped bacteria when enriched in complex nutrient broth (with 10% NaCl). Denitrifiers were recorded abundantly in the sediments, their population decreasing from some 10⁶/g in the surface material to only a few in the subsediment. A total of 16 pure cultures of denitrifying bacteria were isolated from the Suakin-and Thetis-deeps and studied morphologically, physiologically and biochemically. Genetics (molar percentage of guanine plus cytosine, % GC) and numerical taxonomy were included to reveal relationships and improve taxonomic classification. Fifteen isolates were described as Gram-negative, aerobic and facultative anaerobic (with NO ₃ ^- as H⁺-acceptor), polarly flagellated rods (Pseudomonas spp.); one was an inmotile, Gram-positive, facultative anaerobic coccus. None of the 15 Pseudomonas-isolates could be identified with one of the denitrifying species so far described and recognized. The strains should be regarded as hitherto undescribed denitrifying marine bacteria.Supported by the Bundesministerium für Forschung und Technologie, Bonn, Germany (FRG).     

Deponiegas

Investigations were performed concerning whether or not the composition of volatile halogenated carbons in landfill gas (LFG) change when they uncontrolledly leave the aerobic surface of landfill. For this reason, the aerobic biological degradation of chlorofluorocarbons (CFCs), chlorofluorohydrocarbons (HCFCs), and vinyl chloride (VC), which were often measured in LFG of old household-landfills, was investigated. In 1-liter and 126-liter laboratory test reactors, the aerobic milieu conditions of the surface of landfills were simulated. Air with methane and trichlorofluoromethane (R11), dichlorodifluoromethane (R12), and their reductive metabolites dichlorofluoromethane (R21), chlorodifluoromethane (R22), and vinyl chloride (VC) were continuosly pumped through marl and a mixture of biowaste and compost. R21, R22, and VC were degraded, probably cometabolically by methanotrophic bacteria. They have inhibited the methane oxidation. The degradation rates of R21 (only in biowaste and compost) and R22 were up to 2.8 and 10 mg/m³ _material/h, respectively. The maximum degradation rate of VC was 1,3000 mg/m³ _material/h in marl. Under aerobic milieu conditions, a biodegradation of R11 and R12 was not detected.     

Removal of nitrogen from wastewaters by anaerobic ammonium oxidation (ANAMMOX) using granules in upflow reactors

Nitrogen pollution of waters has sometimes caused severe eutrophication, leading to the death of fishes and most aquatic life. There is therefore a need for efficient and cost-effective methods to remove nitrogen from ammonium-rich wastewaters. Anaerobic ammonium oxidation (ANAMMOX) is a promising process to remove nitrogen because this process directly oxidizes ammonium (NH₄ ⁺) to dinitrogen gas (N₂) under anoxic condition. Nonetheless, a challenge of this process is that chemolithoautotrophic Anammox bacteria grow slowly at the beginning, thus resulting in low Anammox biomass and instability of reactors. Such issues can be overcome by granulation of the Anammox sludge. Here, we review the characteristics of the Anammox bacteria, and the formation, structure and flotation of Anammox granules under high hydraulic loadings. We also evaluate the performances of full-scale granular Anammox processes. The major points are: 1) Anammox bacteria secrete a large amount of extracellular polymeric substances (EPS), up to 415 mg g^?1 of volatile suspended solids (VSS), containing many hydrophobic functional groups that facilitate biomass granulation. 2) Granulation enhances the sludge settling property and retention time, which contributes to the extremely high nitrogen removal rate of 77 kg m^?3 d^?1 of Anammox upflow reactors. 3) Flotation of Anammox granules frequently occurs under nitrogen removal rate higher than 10 kg m^?3 d^?1, which is mainly due to the overproduction of EPS under high hydraulic conditions.     

Effects of nitrite on phosphate uptake in anaerobic-oxic process

An anaerobic-oxic (A/O) biological phosphorus removal reactor was operated to study the effect of nitrite on phosphate uptake. The phosphorus uptake profile was determined under different operating conditions. The results indicated that in addition to oxygen and nitrate (DPB_Na, nitrate denitrifying phosphorus removal), to some extent, nitrite could also serve as an electron acceptor to achieve nitrite denitrifying phosphorus removal (DPB_Ni). The quantity and rate of phosphorus uptake of DPB_Ni, however, were evidently lower than that of DPB_Na. The experiment results revealed that nitrite would bring toxic action to phosphate-accumulating organisms (PAOs) when NO₂ ^?-N ? 93.7 mg/L. The nitrite existing in the anoxic reactor made no difference to the quantity and rate of denitrifying phosphorus removal, but it could reduce the consumption of nitrate. Moreover, the data showed that the aerobic phosphate uptake of DPB_Ni was lower than that of anaerobic phosphorus-released sludge in a traditional A/O process. However, there was not much difference between these two kinds of sludge in terms of the total phosphorus uptake quantity and the effluent quality.     

Oxygen transfer in a high compacting multiphasic reactor (H.C.M.R.)

H.C.M.R. is a fluidised multiphasic reactor. In this reactor the rate of oxygen transfer was studied in order to characterise its efficiency as a system of aerobic treatment.

The studied parameters are: The size of the reactor, the volume of the particles, the oxygen transfer rate.

Several experiments were carried out with a variation of the above parameters and the results of the experiments are summarised as follows: The overall mass transfer rate coefficient k ₁ a increases with increasing of upflow gas rate (U_g ); The overall mass transfer rate coefficient k ₁ a decreases with increasing of the inert support volume (V_s); The overall mass transfer rate coefficient values from cylinder shaped particles were higher than those for disc shaped particles.     

Aerobic granulation of pure bacterial strain

The objective of this study is to cultivate aerobic granules by pure bacterial strain, Bacillus thuringiensis, in a sequencing batch reactor. Stable granules sized 2.0–2.2 mm were formed in the reactor after a five-week cultivation. These granules exhibited excellent settling attributes, and degraded phenol at rates of 1.49 and 1.19 g phenol/(g VSS·d) at 250 and 1500 mg/L of phenol concentration, respectively. Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates, and the outer edge of the granule was away from the core regime in the following stage.     

The importance of methane and thiosulfate in the metabolism of the bacterial symbionts of two deep-sea mussels

Undescribed hydrocarbon-seep mussels were collected from the Louisiana Slope, Gulf of Mexico, during March 1986, and the ultrastructure of their gills was examined and compared to Bathymodiolus thermophilus, a mussel collected from the deep-sea hydrothermal vents on the Galápagos Rift in March 1985. These closely related mytilids both contain abundant symbiotic bacteria in their gills. However, the bacteria from the two species are distinctly different in both morphology and biochemistry, and are housed differently within the gills of the two mussels. The symbionts from the seep mussel are larger than the symbionts from B. thermophilus and, unlike the latter, contain stacked intracytoplasmic membranes. In the seep mussel three or fewer symbionts appear to be contained in each host-cell vacuole, while in B. thermophilus there are often more than twenty bacteria visible in a single section through a vacuole. The methanotrophic nature of the seep-mussel symbionts was confirmed in ¹⁴C-methane uptake experiments by the appearance of label in both CO₂ and acid-stable, non-volatile, organic compounds after a 3 h incubation of isolated gill tissue. Furthermore, methane consumption was correlated with methanol dehydrogenase activity in isolated gill tissue. Activity of ribulose-1,5-biphosphate (RuBP) carboxylase and ¹⁴CO₂ assimilation studies indicate the presence of either a second type of symbiont or contaminating bacteria on the gills of freshly captured seep mussels. A reevaluation of the nutrition of the symbionts in B. thermophilus indicates that while the major symbiont is not a methanotroph, its status as a sulfur-oxidizing chemoautotroph, as has been suggested previously, is far from proven.     

Coupled aerobic and anoxic biodegradation for quinoline and nitrogen removals

Quinoline (C₉H₇N) commonly occurs in wastewaters from the chemical, pharmaceutical, and dyeing industries. As quinoline is biodegraded, nitrogen is released as ammonium. Total-N removal requires that the ammonium-N be nitrified and then denitrified. The objective of this study was to couple quinoline biodegradation with total-N removal. In a proof-of-concept step, activated sludge was sequenced from aerobic to anoxic stages. The ammonium nitrogen released from quinoline biodegradation in the aerobic stage was nitrified to nitrate in parallel. Anoxic biodegradation of the aerobic effluent then brought about nitrogen and COD removals through denitrification. Then, simultaneous quinoline biodegradation and total-N removal were demonstrated in a novel airlift internal loop biofilm reactor (AILBR) having aerobic and anoxic zones. Experimental results showed that the AILBR could achieve complete removal of quinoline, 91% COD removal, and 85% total-N removal when glucose added as a supplemental electron donor once nitrate was formed.     

Nitrogen fixation (acetylene reduction) associated with macroalgae in a coral-reef community in the Bahamas

N₂ fixation (C₂H₂ reduction) was associated with several species of macroalgae on a coral reef near Grand Bahama Island. The highest rates were associated with Microdictyon sp. (Chlorophyceae) and Dictyota sp. (Phaeophyceae). Extensive mats of filamentous blue-green algae, not heterotrophic bacteria, were the N₂ fixing agents: in experiments with samples of Microdictyon sp., the activity was lightdependent and not stimulated by organic compounds under either aerobic or anaerobic conditions. Assays in situ, at 20 m depth, and on shipboard, gave similar rates of N₂ fixation; the cyanophytes presumably have pigment adaptations to function in blue light. The maximum rate of N₂ fixation, associated with Microdictyon sp., was 3.8 g N fixed g dry weight^-1 h^-1. Coral-reef communities flourish in nutrientimpoverished waters, and therefore any input of nitrogen is probably important in stabilizing such ecosystems.     

Removal of sulfamethoxazole and trimethoprim from reclaimed water and the biodegradation mechanism

Sulfamethoxazole (SMX) and trimethoprim (TMP) are two critical sulfonamide antibiotics with enhanced persistency that are commonly found in wastewater treatment plants. Recently, more scholars have showed interests in how SMX and TMP antibiotics are biodegraded, which is seldom reported previously. Novel artificial composite soil treatment systems were designed to allow biodegradation to effectively remove adsorbed SMX and TMP from the surface of clay ceramsites. A synergy between sorption and biodegradation improves the removal of SMX and TMP. One highly efficient SMX and TMP degrading bacteria strain, Bacillus subtilis, was isolated from column reactors. In the removal process, this bacteria degrade SMX and TMP to NH ₄ ⁺ , and then further convert NH ₄ ⁺ to NO ₃ ^– in a continuous process. Microbial adaptation time was longer for SMX degradation than for TMP, and SMX was also able to be degraded in aerobic conditions. Importantly, the artificial composite soil treatment system is suitable for application in practical engineering.

Open image in new window

     

Effect of pH on biologic degradation of Microcystis aeruginosa by alga-lysing bacteria in sequencing batch biofilm reactors

In this paper, the effect of pH on biological degradation of Microcystis aeruginosa by alga-lysing bacteria in laboratory-scale sequencing batch biofilm reactors (SBBRs) was investigated. After 10 d filming with waste activated sludge, the biological film could be formed, and the bioreactors in which laid polyolefin resin filler were used to treat algal culture. By comparing the removal efficiency of chlorophyll a at different aerobic time, the optimum time was determined as 5 h. Under pH 6.5, 7.5, and 8.5 conditions, the removal rates of Microcystis aeruginosa were respectively 75.9%, 83.6%, and 78.3% (in term of chlorophyll a), and that of Chemical Oxygen Demand (COD_Mn) were 30.6%, 35.8%, and 33.5%. While the removal efficiencies of ammonia nitrogen (NH ₄ ⁺ - N) were all 100%. It was observed that the sequence of the removal efficiencies of algae, NH ₄ ⁺ - N and organic matter were pH 7.5>pH 8.5>pH 6.5. The results showed that the dominant alga-lysing bacteria in the SBBRs was strain HM-01, which was identified as Bacillus sp. by Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, Basic Local Alignment Search Tool (BLAST) analysis, and comparison with sequences in the GenBank nucleotide database. The algicidal activated substance which HM-01 strain excreted could withstand high temperature and pressure, also had better hydrophily and stronger polarity.     

Advanced nitrogen and phosphorus removal in A2O-BAF system treating low carbon-to-nitrogen ratio domestic wastewater

A laboratory-scale anaerobic-anoxic-aerobic process (A²O) with a small aerobic zone and a bigger anoxic zone and biologic aerated filter (A²O-BAF) system was operated to treat low carbon-to-nitrogen ratio domestic wastewater. The A²O process was employed mainly for organic matter and phosphorus removal, and for denitrification. The BAF was only used for nitrification which coupled with a settling tank Compared with a conventional A²O process, the suspended activated sludge in this A²O-BAF process contained small quantities of nitrifier, but nitrification overwhelmingly conducted in BAF. So the system successfully avoided the contradiction in sludge retention time (SRT) between nitrifying bacteria and phosphorus accumulating organisms (PAOs). Denitrifying phosphorus accumulating organisms (DPAOs) played an important role in removing up to 91% of phosphorus along with nitrogen, which indicated that the suspended activated sludge process presented a good denitrifying phosphorus removal performance. The average removal efficiency of chemical oxygen demand (COD), total nitrogen (TN), total phosphorus (TP), and NH ₄ ⁺ -N were 85.56%, 92.07%, 81.24% and 98.7% respectively. The effluent quality consistently satisfied the national first level A effluent discharge standard of China. The average sludge volume index (SVI) was 85.4 mL·g^?1 additionally, the volume ratio of anaerobic, anoxic and aerobic zone in A²O process was also investigated, and the results demonstrated that the optimum value was 1:6:2.     

Treatment of swine wastewater in aerobic granular reactors: comparison of different seed granules as factors

The granulation process, physic-chemical properties, pollution removal ability and bacterial communities of aerobic granules with different feed-wastewater (synthetic wastewater, R1; swine wastewater, R2), and the change trend of some parameters of two types of granules in long-term operated reactors treating swine wastewater were investigated in this experiment. The result indicated that aerobic granulation with the synthetic wastewater had a faster rate compared with swine wastewater and that full granulation in R1 and R2 was reached on the 30th day and 39th day, respectively. However, although the feed wastewater also had an obvious effect on the biomass fraction and extracellular polymeric substances of the aerobic granules during the granulation process, these properties remained at a similar level after long-term operation. Moreover, a similar increasing trend could also be observed in terms of the nitrogen removal efficiencies of the aerobic granules in both reactors, and the average specific removal rates of the organics and ammonia nitrogen at the steady-state stage were 35.33 mg·g⁻¹ VSS and 51.46 mg·g⁻¹ VSS for R1, and 35.47 mg·g⁻¹ VSS and 51.72 mg·g⁻¹ VSS for R2, respectively. In addition, a shift in the bacterial diversity occurred in the granulation process, whereas bacterial communities in the aerobic granular reactor were not affected by the seed granules after long-term operation.     

Structure and formation of anoxic granular sludge—A string-bag hypothesis

Anoxic granular sludge was developed in a laboratory-scale sequencing batch reactor which was fed with sodium acetate and sodium nitrate as electron donor and accepter. The sludge in the reactor was almost granulated after approximately 90 days of cultivation. In the present study, a detailed examination of surface morphology and internal structure of anoxic granular sludge was conducted using scanning electron microscope. It showed that the bacteria inside the granules had a uniform, coccus-like shape. By contrast, filamentous bacteria were predominant outside the granules. These bacteria were woven and had wrapped the coccus bacteria together to form granules. The small amounts of DO in the liquid bulk promoted the growth of filamentous bacteria on the surface of the granules. A string-bag hypothesis was proposed to elucidate the structure and formation of the anoxic granular sludge. It suggested that micro-aeration could be a method to promote granulation in practical anoxic treatment systems.     

Sediment bacteria in the deep Norwegian Sea

During a joint French-Swedish expedition (NORBI) in July-August, 1975, in the deep Norwegian Sea, the density of aerobic, heterotrophic sediment bacteria was determined by viable counts at 9 stations. The numbers rarely exceeded 10³ ml^-1 wet sediment. ATP measurements, with a limit of detectability of 5 ng ml^-1 wet sediment, gave constantly negative results. One third of the strains isolated at 5°C were psychrophiles. The ability to degrade lipids, proteins, starch, and chitin, probably due to maximum biosynthesis of the exoenzymes, was correlated with the optimum temperature for growth. There was a strong positive correlation (P<0.001) between optimum temperature for the isolates and the number of different macromolecules attacked. Lipolytic activity was found to predominate. Differences in the bacterial composition between the Norwegian and Lofoten, Spitsbergen, and Greenland deep basins are discussed in relation to known differences in their faunal density and hydrography.     

Anaerobic treatment of fresh leachate from a municipal solid waste incinerator by upflow blanket filter reactor

This paper describes the feasibility of fresh leachate treatment by an upflow blanket filter (UBF). Through dilution and partial effluent recycling, the organic loading rates increased from 0.51 to 14.56 kg COD/(m³·d), meanwhile the corresponding hydraulic retention time decreased from 9.0 to 3.6 d. The reactor was able to achieve steady-state within 80 d. Based on the distribution of COD fluxes in the process, it was concluded that anabolism was the main pathway of COD removal in the initial phase (1–33 d), accounting for 57%–85% of total COD removed. As the anaerobic consortium of bacteria reached steady-state (after 70–86 d), the majority of COD removed was transformed into methane, because the specific methane yield was close to the theoretical value (0.36 L CH₄/(g COD_deg)).     

Effects of anaerobiosis on root metabolism of Zostera marina (eelgrass): implications for survival in reducing sediments

The temperate seagrass Zostera marina L. typically grows in highly reducing sediments. Photosynthesis-mediated O₂ supplied to below-ground tissues sustains aerobic respiration during photosynthetic periods. Roots, however, experience daily periods of anoxia and/or hypoxia at night and under conditions that reduce photosynthesis. Rhizosphere cores of Z. marina were collected in August 1984 from Great Harbor, Massachusetts, USA. We examined short-term anaerobic metabolism of [U-¹⁴C]sucrose in excised roots and roots of intact plants. Under anaerobic conditions roots showed appreciable labeling of CO₂, ethanol and lactate, and slight labeling of alanine and other metabolites. Over 95% of the ¹⁴C-ethanol was recovered in the root exudate. Release of other metabolites from the roots was minimal. Ethanol was also released from hypoxic/anoxic roots of intact plants and none of this ethanol was transported to the shoot under any experimental conditions. Loss of ethanol from roots prevented tissue levels of this phytotoxin from increasing during anaerobiosis despite increased synthesis of ethanol. Anaerobic metabolism of [U-¹⁴C]glutamate in excised roots led to appreciable labelling of -aminobutyrate, which was known to accumulate in eelgrass roots. Roots recovered to fully aerobic metabolism within 4 h after re-establishment of aerobic conditions. The contributions of these root metabolic responses to the ability of Z. marina to grow in reducing marine sediments are related to light-regulated interactions of shoots and roots that likely dictate depth penetration, distribution and ecological success of eelgrass.