Quorum sensing responses of activated sludge to free nitrous acid: Zoogloea deformation, AHL redistribution, and microbiota acclimatization.

Free nitrous acid (FNA) has been widely employed for improvement of wastewater management by altering sludge characteristic and function based on its polymer lysing and biocidal capacity. Sludge characteristic and function are commonly considered as the joint consequence of microbial individual behaviors and quorum sensing (QS) involved collective behaviours, but the role of the latter in FNA treatment was still as-yet-unidentified and addressed in this research. The results of sludge morphology and component characterized FNA-induced zoogloea deformation, including inner cell exposure, half of extracellular polymeric substances (EPS) reduction and adsorption site depletion. During zoogloea deformation, four acyl-homoserine lactones (AHLs), including C4-HSL, C8-HSL, C10-HSL and C12-HSL, transferred inward of microbiota, and their total contents reduced by 66% because of depressed signal production, augmented decomposer and recognition. Transcriptome analysis revealed that differentially expressed QS driven by AHL redistribution facilitated microbiota acclimatization including cellular motility and hydrolase synthesis for EPS consumption. Boosted motility may favor escaping from stress spot and moderating intercellular acidity based on cell motility test. Feasible EPS consumption provided nutrition for heterotrophic metabolisms testified by pure culture with EPS as sole nutrition. Our work thus comprehensively revealed QS behaviours responding to FNA and deepened the understanding to FNA treatment performance in wastewater management.

Production efficiency and properties of poly(3hydroxybutyrate-co-3hydroxyvalerate) generated via a robust bacterial consortium dominated by Zoogloea sp. using acidified discarded fruit juices as carbon source.

In the current study, a mixed microbial culture (MMC) of polyhydroxyalkanoates (PHAs) producers was developed under nutrient stress and was assessed as biocatalyst for the production of high-yielding PHAs from fermented (acidified) discarded fruit juices (DFJ). The structure of the MMC was analyzed periodically to determine its microbial dynamics, revealing that Zoogloae sp. dominated throughout the operation of the system. The efficiency of PHAs production from the MMC was further optimized in batch mode by altering the ratio of C to N, the ratio of carbon sources (propionate and butyrate), and the initial pH, and subsequently different fermentation mixtures of acidified DFJ were assessed as substrates at optimal conditions. Upon solvent extraction, the properties of recovered PHAs were analyzed, showing that in all cases P(3HB-co-3HV) was produced, with Tm ranging from 90.5 to 168.8 °C, and maximum obtained yields 54.61 ± 4.31 % and 43.27 ± 2.13 %, from synthetic substrates and DFJ, respectively. Overall, it was shown that the developed MMC can be efficiently applied as biocatalyst for the exploitation of sugary wastewaters, such as DFJ, towards bio-based and biodegradable plastics bearing the required properties to substitute fossil plastics, into the concept of a circular economy.

Stability of pure oxygen aeration-activated sludge system under non-steady food-to-microorganism ratio conditions during petrochemical wastewater treatment.

This study investigates the stability of a pure oxygen aeration-activated sludge system for petrochemical wastewater treatment under high organic concentration and non-steady food-to-microorganism (F/M) ratio conditions. Sludge settling characteristics maintained relatively stable conditions with an F/M ratio variation from 0.15 ± 0.04 to 0.33 ± 0.07 kg COD/kg MLSS⋅d, while the excess F/M ratio (0.44 ± 0.16 kg COD/kg MLSS⋅d) resulted in deterioration of the organic removal and sludge-water separation performances. Loosely bound extracellular polymeric substances (EPS) showed more significant effect on sludge settleability than the tightly bound EPS. The genus Hydrogenophaga was related to organic removal performance, while Zoogloea and Chitinophaga were related to the effluent quality of suspended solids. The excess F/M ratio also caused an increase in Zoogloea and Chitinophaga, whereas the toxicity of petrochemical wastewater resulted in decreased abundance of Hydrogenophaga. These changes caused deterioration of the organic removal and sludge-water separation performances.

Genome analysis provides insights into microaerobic toluene-degradation pathway of Zoogloea oleivorans BucT.

Zoogloea oleivorans, capable of using toluene as a sole source of carbon and energy, was earlier found to be an active degrader under microaerobic conditions in aquifer samples. To uncover the genetic background of the ability of microaerobic toluene degradation in Z. oleivorans, the whole-genome sequence of the type strain BucT was revealed. Metatranscriptomic sequence reads, originated from a previous SIP study on microaerobic toluene degradation, were mapped on the genome. The genome (5.68 Mb) had a mean G + C content of 62.5%, 5005 protein coding gene sequences and 80 RNA genes. Annotation predicted that 66 genes were involved in the metabolism of aromatic compounds. Genome analysis revealed the presence of a cluster with genes coding for a multicomponent phenol-hydroxylase system and a complete catechol meta-cleavage pathway. Another cluster flanked by mobile-element protein coding genes coded a partial catechol meta-cleavage pathway including a subfamily I.2.C-type extradiol dioxygenase. Analysis of metatranscriptomic data of a microaerobic toluene-degrading enrichment, containing Z . oleivorans as an active-toluene degrader revealed that a toluene dioxygenase-like enzyme was responsible for the ring-hydroxylation, while enzymes of the partial catechol meta-cleavage pathway coding cluster were responsible for further degradation of the aromatic ring under microaerobic conditions. This further advances our understanding of aromatic hydrocarbon degradation between fully oxic and strictly anoxic conditions.

Effects of DO levels on surface force, cell membrane properties and microbial community dynamics of activated sludge.

In this paper, we employ atomic force microscopy (AFM), fluorescence recovery after photobleaching (FRAP) technique, phospholipid fatty acids (PLFA) and MiSeq analysis to study the effects of traditional dissolved oxygen (DO) levels (0.71-1.32mg/L, 2.13-3.02mg/L and 4.31-5.16mg/L) on surface force, cell membrane properties and microbial community dynamics of activated sludge. Results showed that low DO level enhanced the surface force and roughness of activated sludge; the medium DO level decreased cell membrane fluidity by reducing the synthesis of branched fatty acids in the cell membrane; high DO level resulted in the highest protein content in the effluent by EEM scanning. Abundance of Micropruina, Zoogloea and Nakamurella increased and Paracoccus and Rudaea decreased with the increase of DO levels. RDA analysis suggested that saturated fatty acids (SFA), anteiso-fatty acids (AFA) and iso-fatty acids (IFA) were closely related to effluent quality as well as some genera.

Nitrogen Removal Characteristics of a Newly Isolated Indigenous Aerobic Denitrifier from Oligotrophic Drinking Water Reservoir, Zoogloea sp. N299.

Nitrogen is considered to be one of the most widespread pollutants leading to eutrophication of freshwater ecosystems, especially in drinking water reservoirs. In this study, an oligotrophic aerobic denitrifier was isolated from drinking water reservoir sediment. Nitrogen removal performance was explored. The strain was identified by 16S rRNA gene sequence analysis as Zoogloea sp. N299. This species exhibits a periplasmic nitrate reductase gene (napA). Its specific growth rate was 0.22 h-1. Obvious denitrification and perfect nitrogen removal performances occurred when cultured in nitrate and nitrite mediums, at rates of 75.53%±1.69% and 58.65%±0.61%, respectively. The ammonia removal rate reached 44.12%±1.61% in ammonia medium. Zoogloea sp. N299 was inoculated into sterilized and unsterilized reservoir source waters with a dissolved oxygen level of 5-9 mg/L, pH 8-9, and C/N 1.14:1. The total nitrogen removal rate reached 46.41%±3.17% (sterilized) and 44.88%±4.31% (unsterilized). The cell optical density suggested the strain could survive in oligotrophic drinking water reservoir water conditions and perform nitrogen removal. Sodium acetate was the most favorable carbon source for nitrogen removal by strain N299 (p<0.05). High C/N was beneficial for nitrate reduction (p<0.05). The nitrate removal efficiencies showed no significant differences among the tested inoculums dosage (p>0.05). Furthermore, strain N299 could efficiently remove nitrate at neutral and slightly alkaline and low temperature conditions. These results, therefore, demonstrate that Zoogloea sp. N299 has high removal characteristics, and can be used as a nitrogen removal microbial inoculum with simultaneous aerobic nitrification and denitrification in a micro-polluted reservoir water ecosystem.

In situ profiling of microbial communities in full-scale aerobic sequencing batch reactors treating winery waste in australia.

On-site aerobic sequencing batch reactor (SBR) treatment plants are implemented in many Australian wineries to treat the large volumes of associated wastewater they generate. Yet very little is known about their microbiology. This paper represents the first attempt to analyze the communities of three such systems sampled during both vintage and nonvintage operational periods using molecular methods. Alphaproteobacterial tetrad forming organisms (TFO) related to members of the genus Defluviicoccus and Amaricoccus dominated all three systems in both operational periods. Candidatus 'Alysiosphaera europaea' and Zoogloea were codominant in two communities. Production of high levels of exocellular capsular material by Zoogloea and Amaricoccus is thought to explain the poor settleability of solids in one of these plants. The dominance of these organisms is thought to result from the high COD to N/P ratios that characterize winery wastes, and it is suggested that manipulating this ratio with nutrient dosing may help control the problems they cause.

Effect of temperature and cycle length on microbial competition in PHB-producing sequencing batch reactor.

The impact of temperature and cycle length on microbial competition between polyhydroxybutyrate (PHB)-producing populations enriched in feast-famine sequencing batch reactors (SBRs) was investigated at temperatures of 20 °C and 30 °C, and in a cycle length range of 1-18 h. In this study, the microbial community structure of the PHB-producing enrichments was found to be strongly dependent on temperature, but not on cycle length. Zoogloea and Plasticicumulans acidivorans dominated the SBRs operated at 20 °C and 30 °C, respectively. Both enrichments accumulated PHB more than 75% of cell dry weight. Short-term temperature change experiments revealed that P. acidivorans was more temperature sensitive as compared with Zoogloea. This is particularly true for the PHB degradation, resulting in incomplete PHB degradation in P. acidivorans at 20 °C. Incomplete PHB degradation limited biomass growth and allowed Zoogloea to outcompete P. acidivorans. The PHB content at the end of the feast phase correlated well with the cycle length at a constant solid retention time (SRT). These results suggest that to establish enrichment with the capacity to store a high fraction of PHB, the number of cycles per SRT should be minimized independent of the temperature.

[Selection and kinetic mechanism of psychrotrophs in low temperature wastewater treatment].

A study was carried out taking low temperature domestic wastewater as target in laboratory, the composition of bacteria in activated sludge was analyzed and the degradation kinetics of organics was investigated. Six preponderant psychrotrophs were selected out, belonging to Zoogloea, Aeromonas, Flavobacterium, Micrococcus, Bacillus and Pseudomonaus, respectively. Results show that TTC-dehydrogenase activity of the psychrotroph is 25.44 mg/(L x h) which is 20.5 times more than that of ordinary activated sludge at 4 degrees C. The efficiency of COD biodegradation by psychrotrophs at low and normal temperature is 80.9% and 73.4%, respectively. Mesophilic bacteria almost lost their activity at low temperature. Kinetic analysis shows that biodegradation of organics by psychrotrophs at low and mesothermal temperature as well as mesophilic bacteria at mesothermal temperature are in accordance with the model of first-order reaction. Psychrotrophs, which assure the removal efficiency of organic pollutants at low and normal temperature, could keep the uniform reaction velocity as the mesophilic bacteria and also adapt wide ecological amplitude of temperature. Efficient psychrotrophs were immobilized on soft polyurethane foams which acted as carriers in the experiment at 4 degrees C, as a result of that, the removal efficiency of COD was increased about 18% higher than that of mobilized ones at low temperature, the biodegradation kinetics of COD by immobilized psychrotrophs also followed the first-order reaction model. With glucose in water as the source of nutriment, the reaction velocity of immobilized bacteria is 2.35 times higher than that of the mobilized ones. By the immobilized psychrotrophs biodegradation of varied nutriment, the effluent could achieve the first-degree B of pollutants discharge standard for municipal wastewater treatment plant.

[Biodegradation of pyridine by Shinella zoogloeoides BC026].

A bacterial strain BC026 capable of utilizing pyridine as its sole source of carbon and nitrogen was isolated from the activated sludge in a coking wastewater treatment plant. The bacterium featured flocculability and antibiotic resistance to kanamycin, ampicillin and spectinomycine. It could grow well in Ashby nitrogen free culture medium. The strain was identified as Shinella zoogloeoides according to the results of 16S rRNA sequence analysis and Biolog microbial identification system. The experiments of pyridine biodegradation by the pure culture showed that pyridine of 400 mg/L could be degraded completely in 17 h under the condition of inoculum 0.1 g/L, 30 degrees C, 180 r/min and pH 7. BC026 could keep high degradative activity in mineral salt medium containing pyridine with a concentration ranging from 99 mg/L to 1 806 mg/L. Higher initial concentration of pyridine caused repression on BC026 to a certain extent, however, the degradation rate became faster after the strain had been accommodated. The optimal conditions for the degradation were 30-35 degrees C and pH 8. The research on metabolic pathway of pyridine by BC026 indicated that the first step of pyridine degradation was C-N bonds cleavage, generating NH4+ and glutaraldehyde. Then glutaraldehyde was oxidized into glutaric acid, and finally into CO2 and H2O. 59.5% nitrogen from pyridine was transferred into ammonium in the whole degradation.

Physicochemical properties of the exopolysaccharides produced by marine bacterium Zoogloea sp. KCCM10036.

The physicochemical properties of the exopolysaccharide (EPS) produced by marine bacterium Zoogloea sp. KCCM10036 were investigated. Two types of isolated EPSs were shown to have average relative molecular masses (Mr) of 4.07 x 10(6) of CBP (cell-bound polysaccharide) and 3.43 x 10(6) of WSP (water-soluble polysaccharide), respectively. When the CBP was utilized as an emulsifier, it stabilized the emulsion for up to 148 h. Compared with other commercially available hydrocolloids such as xanthan gum, the Tween series, and Triton, the CBP showed much better emulsifying capability on a water-in-oil system. Phase separation occurred in the Tween series after 24 h, whereas the emulsion was better stabilized by the CBP. The CBP thus has potential as an emulsifying agent in commercial emulsions. The flocculating activity was also greatest at 0.01% (w/v) and decreased at higher concentrations than the optimized concentration of the WSP and CBP. The results also showed that both types of exopolysaccharides from Zoogloea sp. had excellent flocculating activity.

[Degradation and kinetics of pyrene and benzo [a] pyrene by three bacteria in contaminated soil].

Degradation kinetics of pyrene and benzo [a] pyrene (BaP) in soil by three individual strains of bacteria were examined using the Michaelis-Menton and Monod dynamic models. There were significant differences occurred in the degradation rates of pyrene and BaP by the three individual strains. When the initial concentrations of pyrene or BaP was 50 mg/kg in soil, the degradation rates of pyrene by Bacillus sp. SB02, Zoogloea sp.SB09, and Flavobacterium sp.SB10 in 42 days were 42.69%, 32.88% and 25.07% respectively for pyrene. Similarly, degradation rates of BaP by Bacillus sp. SB02, Zoogloea sp. SB09, and Flavobacterium sp. SB10 in 42 days were 33.04%, 25.39% and 22.02% respectively for BaP. In addition, there also were significant differences occurred in the degradation rates per day of pyrene and BaP by the three individual strains. The most rapid degradation of pyrene and BaP by Bacillus sp. SB02 occurred in one week, and 20.88% of pyrene and 12.6% of BaP were degraded. However, Zoogloea sp. SB09 had a slower degradation, while Flavobacterium sp. SB10 had the slowest degradation.

[Non-filamentous sludge bulking caused by low concentration of dissolved oxygen and phosphorus deficiency and its control].

Non-filamentous sludge bulking occurred during sludge acclimation in lab-scale bio-reactors and the characteristics, properties and degrading pollutants capability of non-filamentous sludge were analyzed. Both low DO concentration (0 - 0.7 mg/L) and low ratio of P/BOD5 (0.78/100) resulted in excessive proliferation of viscous zoogloea and sludge bulking. SVI was directly proportional to extracellular polysaccharides and was inversely proportional to hydrophobicity of sludge. The sludge bulking could be effectively controlled by increasing DO concentration and P/BOD5 ratio. Besides, adding some porous padding into viscous bulking sludge was a useful method which might avoid the deterioration of sludge settlement and maintain good performance in the system. The viscous bulking sludge could not be controlled by addition of NaC1O.

Impact of septic compounds and operational conditions on the microbiology of an activated sludge system.

In activated sludge (AS) biotreatment, septic compounds such as volatile organic acids and reduced sulphur compounds have been frequently cited as a major cause of Thiothrix and Type 021N filamentous bulking. These filaments are common in Canadian pulp and paper biotreatment systems, where they cause settling problems in secondary clarifiers. We conducted a 14-week study of a TMP/newsprint mill effluent to characterize the septic compounds entering the biotreatment, and to determine correlations with AS biomass characteristics and biotreatment operating parameters. A significant correlation was found between the sludge volume index, the abundance of Type 021N, and the propionic acid (PA) concentration in the primary clarified effluent. PA also induced a significant change in the flocculating bacteria size distribution determined by digital imaging. Consequently, the correlation observed between PA and Type 021N bulking is an indirect effect of inhibition of floc-forming microorganisms, giving a competitive advantage to filaments.

Degradation mechanisms of benzo[a]pyrene and its accumulated metabolites by biodegradation combined with chemical oxidation.

A high degradation extent of benzo[a]pyrene (BaP) should not be considered as the sole desirable criterion for the bioremediation of BaP-contaminated soils because some of its accumulated metabolites still have severe health risks to human. Two main metabolites of BaP, benzo[a]pyrene-1,6-quinone (BP1,6-quinone) and 3-hydroxybenzo[a]pyrene (3-OHBP) were identified by high performance liquid chromatography (HPLC) with standards. This study was the first time that degradation of both BaP and the two metabolites was carried out by chemical oxidation and biodegradation. Three main phases during the whole degradation process were proposed. Hydrogen peroxide-zinc (H(2)O(2)-Zn), the fungus - Aspergillus niger and the bacteria - Zoogloea sp. played an important role in the different phases. The degradation parameters of the system were also optimized, and the results showed that the effect of degradation was the best when fungus-bacteria combined with H(2)O(2)-Zn, the concentration range of BaP in the cultures was 30-120mg/l, the initial pH of the cultures was 6.0. However, as co-metabolites, phenanthrene significant inhibited the degradation of BaP. This combined degradation system compared with the conventional method of degradation by domestic fungus only, enhanced the degradation extent of BaP by more than 20% on the 12d. The highest accumulation of BP1,6-quinone and 3-OHBP were reduced by nearly 10% in the degradation experiments, which further proved that the combined degradation system was more effective as far as joint toxicity of BaP and its metabolites are concerned.

Isolation and characterization of heterotrophic bacteria able to grow aerobically with quaternary ammonium alcohols as sole source of carbon and nitrogen.

The quaternary ammonium alcohols (QAAs) 2,3-dihydroxypropyl-trimethyl-ammonium (TM), dimethyl-diethanol-ammonium (DM) and methyl-triethanol-ammonium (MM) are hydrolysis products of their parent esterquat surfactants, which are widely used as softeners in fabric care. We isolated several bacteria growing with QAAs as the sole source of carbon and nitrogen. The strains were compared with a previously isolated TM-degrading bacterium, which was identified as a representative of the species Pseudomonas putida (Syst. Appl. Microbiol. 24 (2001) 252). Two bacteria were isolated with DM, referred to as strains DM 1 and DM 2, respectively. Based on 16S-rDNA analysis, they provided 97% (DM 1) and 98% (DM 2) identities to the closest related strain Zoogloea ramigera Itzigsohn 1868AL. Both strains were long, slim, motile rods but only DM 1 showed the floc forming activity, which is typical for representatives of the genus Zoogloea. Using MM we isolated a Gram-negative, non-motile rod referred to as strain MM 1. The 16S-rDNA sequence of the isolated bacterium revealed 94% identities (best match) to Rhodobacter sphaeroides only. The strains MM 1 and DM 1 exclusively grew with the QAA which was used for their isolation. DM 2 was also utilizing TM as sole source of carbon and nitrogen. However, all of the isolated bacteria were growing with the natural and structurally related compound choline.

Soft gel medium solidified with gellan gum for preliminary screening for root-associating, free-living nitrogen-fixing bacteria inhabiting the rhizoplane of plants.

For preliminary screening for and characterization of free-living nitrogen-fixing bacteria from rhizoplane microflora, we used Winogradsky's mineral mixture-based nitrogen-free medium solidified with 0.3% gellan gum. The soft gel medium enabled some reference and wild free-living nitrogen-fixing bacteria to grow in characteristic colonies, including their reaction to oxygen and their motility change. Gellan gum is thus likely to be a better gel matrix than agarose for the investigation of root-associating, free-living nitrogen-fixing bacteria to identify their characteristic behaviors.

[The degradation of phenanthrene and pyrene contaminated soil with immobilized technique].

The phenanthrene and pyrene contaminated soil was degradaded by immobilized-imbedding the selected Zoogloea sp. The degradation rate was determined under various amounts of inoculation and concentration, and the result showed that there was the best degradation rate at 5%. After 168 h, the degradation rate of immobilized Zoogloea sp. to phenanthrene and pyrene was 84.89% and 76.94% respectively. While, the degradation rate of the native bacterium was only 27.85% and 19.65% at the same condition. So the Zoogloea sp. had the better degradation ability. And, Zoogloea sp. distributing shape in immobilized carrier was studied with electric scan-photo, it inferred that immobilized bacterium had the better degradation rate.

Identification and characterization of poly-3-hydroxybutyrate granule-associated protein, PGA12 and PGA16 in Zoogloea ramigera I-16-M.

Poly-3-hydroxybutyrate (PHB) granules of Zoogloea ramigera I-16-M contained two major PHB granule-associated proteins (PGA12 and PGA16) as revealed by sodium dodecyl sulfate-polyacrylamide gel elecrophoresis. N-terminal amino acid sequences of these proteins were determined. The genes encoding these proteins were cloned and sequenced. The structural genes of PGA12 and PGA16 were 351 and 447 bp long, which encode polypeptides with deduced molecular masses of 12.3 and 16.0 kDa, respectively. PGA12 and PGA16 were expressed in Escherichia coli. PHB granules were isolated from cells of recombinant strains of E. coli JM109, which harbored and expressed the PHB-synthetic genes of Ralstonia eutropha H16 and PGA12 or PGA16. These PHB granules contained PGA12 or PGA16 as a major protein. The presence of pga12 or pga16 did not affect the amount of PHB synthesized in E. coli. PGA12 and PGA16 bound to crystalline and amorphous PHB granules.

[Degradation of phenanthrene and pyrene in contaminated soil by immobilized Zoogloea sp. and Fusarium sp].

Immobilized with PVA, sodium alginate and activated carbon, both Zoogloea sp. and Fusarium sp. strains could degrade phenanthrene and pyrene efficiently. The optimal carrier was made of 100 rho.g-1 L PVA, 5 sodium alginate rho.g-1 L and 50 activated carbon rho.g-1 L. The degradation rates of phenanthrene and pyrene in 10 days were 87.48% and 75.34% by the immobilized bacterium, 37.04% and 20.85% higher than those by the free bacterium, and the rates in 15 days were 84.36% and 74.87% by the immobilized fungus, 5.35% and 11.23% higher than those by the free fungus.