Characterization and genome analysis of Acinetobacter oleivorans S4 as an efficient hydrocarbon-degrading and plant-growth-promoting rhizobacterium.

Plant-growth-promoting rhizobacteria (PGPR) have received increasing attention for assisting phytoremediation. However, the effect of PGPR on total petroleum hydrocarbon (TPH) degradation and plant growth promotion and its underlying mechanism is not well understood. In this study, phenotypic analysis and whole genome sequencing were conducted to comprehensively characterize a newly isolated rhizobacterium strain S4, which was identified as Acinetobacter oleivorans, from a TPH-contaminated soil. The strain degraded 62.5% of initially spiked diesel (1%) in minimal media within six days and utilized n-alkanes with a wide range of chain length (i.e., C12 to C40). In addition, the strain showed phenotypic traits beneficial to plant growth, including siderophore production, indole-3-acetic acid synthesis and phosphate solubilization. Potential metabolic pathways and genes encoding proteins responsible for the phenotypic traits were identified. In a real TPH-contaminated soil, inoculation of Acinetobacter oleivorans S4 significantly enhanced the growth of tall fescue relative to the soil without inoculation. In contrast, inoculation of Bacillus sp. Z7, a hydrocarbon-degrading strain, showed a negligible effect on the growth of tall fescue. The removal efficiency of TPH with inoculation of Acinetobacter oleivorans S4 was significantly higher than those without inoculation or inoculation of Bacillus sp. Z7. These results suggested that traits of PGPR beneficial to plant growth are critical to assist phytoremediation. Furthermore, heavy metal resistance genes and benzoate and phenol degradation genes were found in the genome of Acinetobacter oleivorans S4, suggesting its application potential in broad scenarios.

A sustainable approach of turning potato waste towards bioethanol production using indigenous microbes of Himachal Pradesh, India.

Potato peel waste is one of the zero-value wastes with the potential of bioethanol production through the Waste to Energy (WtE) approach. The newly isolated, phenotypically characterized, and molecular identified high-altitude strain, B. amyloliquefaciens, shown promising starch hydrolysis (12.06 g/L reducing sugars) over acid hydrolysis and is capable of working at 30-50 °C and pH 6.0-8.0. The ethanol production by Acinetobacter sp. (a newly isolated, phenotypically characterized, molecular identified) has been modelled and optimized through the central composite design of response surface methodology by taking the fermentation variables as input variables and ethanol yield as the output variable. The ethanol production by Acinetobacter sp. showcased a non-linear relationship of fermentation variables with the ethanol yield (5.83 g/L) with a 99.11% desirability function (R2) and 97.50 adj. R2 values. Optimal fermentation variables of 38.8% substrate concentration, 7% inoculum, pH 5.45 have been utilized for bioethanol production in 55.27 h at 27 °C. Overall, the present study evaluated the efficiency of newly isolated, indigenous extremophilic microbes of The Himalayan region in sustainable bioethanol production from zero-value waste "Potato peel waste" through the WtE approach. Moreover, the present study introduces the promising, unexplored extremophilic microbial strains with the starch-hydrolyzing and fermentation capabilities to bioethanol biorefinery.

Bioremediation of gaseous methyl tert-butyl ether by combination of sulfuric acid modified bagasse activated carbon-bone biochar beads and Acinetobacter indicus screened from petroleum contaminated soil.

Combination of sulfuric acid modified bagasse activated carbon-bone biochar beads and Acinetobacter indicus screened from petroleum contaminated soil was the best condition for gaseous methyl tert-butyl ether (MTBE) removal. It was found that H2SO4 modified bagasse AC in powder form had higher adsorption capacity (989.33 mg g-1) than that in bead form (1.94 mg g-1). In addition, bone biochar in powder form (3.51 mg g-1) also had higher adsorption capacity than that in bead form (1.63 mg g-1). This was the fact that material beads contained high moisture content that inhibited the penetration of gaseous MTBE into the material. And a mixed material of H2SO4 modified bagasse AC-bone biochar beads had the highest adsorption capacity (2.22 mg g-1) compared to individual H2SO4 modified bagasse AC beads (1.94 mg g-1) and bone biochar beads (1.63 mg g-1) due to a mixed material had more rough surface and high surface area on its material. So, gaseous MTBE can penetrate through this material more easily. Although the maximum adsorption capacity of H2SO4 modified bagasse AC in powder form was the highest but microorganism cannot sustain and survive in this form for a long time. Therefore, the material beads were more suitable for microorganism to grow and degrade gaseous MTBE. Microorganism can degrade MTBE and caused no secondary wastes. Moreover, A. indicus was a novel strain for MTBE removal that has not been previously reported. Therefore, a combination of A. indicus-mixed material beads was a good choice for MTBE removal in a biofilter system.

[Simultaneous Nitrogen and Phosphorus Removal and Kinetics by the Heterotrophic Nitrifying Bacterium Acinetobacter junii NP1].

Due to the problems of traditional biological nitrogen and phosphorus removal, including long process duration and high infrastructural and operational costs, the simultaneous nitrogen and phosphorus removal capabilities, influencing factors and kinetic characteristics were systematically studied using the heterotrophic nitrifier Acinetobacter junii NP1 which possesses efficient simultaneous nitrogen and phosphorus removal ability. The results showed that strain NP1 exhibited efficient heterotrophic nitrification ability with a maximum ammonia removal rate of 99.12%. Furthermore, only small amounts of nitrification intermediates were accumulated during the reaction process. Strain NP1 also adapted well to higher ammonia nitrogen loading. In addition, strain NP1 had efficient aerobic denitrification characteristics, and could utilize nitrite and nitrate for growth and metabolism, achieving a maximum removal rate of 91.40% and 95.10%, respectively. The heterotrophic nitrification process of strain NP1 was accompanied by simultaneous phosphorus accumulation, and the appropriate ratio of nitrogen to phosphorus was beneficial for the simultaneous removal of nitrogen and phosphorus. When the ratio of nitrogen to phosphorus was 5:1, the maximum ammonia nitrogen and phosphate removal rates reached 99.21% and 88.35%, respectively. The bacterial growth process of stain NP1 matched the Logistic model (R2>0.99), and the nitrogen and phosphate degradation conformed to the Compertz model (R2>0.99). The maximum conversion rates of nitrogen and phosphate (Rm) obtained by model fitting were in the order ammonia>nitrate>nitrite, and lag time (t0) was in the order nitrate>nitrite>ammonia. According to the analysis of the degradation kinetics of the matrix and the removal rate of nitrogen and phosphorus, the optimal conditions were found to be sodium succinate, C/N=10, T=30℃, and r=160 r·min-1.

Environmental fungi and bacteria facilitate lecithin decomposition and the transformation of phosphorus to apatite.

Organophosphorus compounds (OP) are stable P source in nature, and can increase eutrophication risk in waterbodies. Lecithin was the most difficult OP to be broken down. In this study, two typical phosphate-solubilizing microorganisms, Aspergillus niger and Acinetobacter sp., were applied to evaluate their ability to decompose both inorganic phosphates and lecithin. A. niger and Acinetobacter sp. could solubilize calcium phosphates by secreting various organic acids, e.g., oxalic and formic acids. The fungus, A. niger, shows significantly higher ability of solubilizing these inorganic phosphates than Acinetobacter sp., primarily due to its secretion of abundant oxalic acid. However, the bacterium, Acinetobacter sp., could secrete more acid phosphatase than A. niger for lecithin decomposition, i.e., 9300 vs. 8500 µmol L-1 h-1. Moreover, after addition of CaCl2, the released P from lecithin was transformed to stable chlorapatite in the medium. To the contrast, Ca cations inclined to form calcium oxalate (rather than stable phosphate mineral) after the incubation of A. niger, as it induced relatively acidic environment after breaking down lecithin. Therefore, this work sheds light on the bright future of applying bacteria and Ca cations in OP pollutant management.

Bioaugmentation coupled with phytoremediation for the removal of phenolic compounds and color from treated palm oil mill effluent.

The potential for coupling bioaugmentation with phytoremediation to simultaneously treat and utilize treated palm oil mill effluent (TPOME) in animal feed production was determined from a reduction in phenolic compounds and color in soil leachates, as well as from an increased yield of pasture grass. Two phenol-degrading bacteria-Methylobacterium sp. NP3 and Acinetobacter sp. PK1-were inoculated into the Brachiaria humidicola rhizosphere before the application of TPOME. A pot study showed that the soil with both grass and inoculated bacteria had the highest dephenolization and decolorization efficiencies, with a maximum capability of removing 70% from 587 mg total phenolic compounds added and 73% from 4438 color units during ten TPOME application cycles. The results corresponded to increases in the number of phenol-degrading bacteria and the grass yield. In a field study, this treatment was able to remove 46% from 21,453 mg total phenolic compounds added, with a maximum color removal efficiency of 52% from 5105 color units, while the uninoculated plots removed about 24-39% and 29-46% of phenolic compounds and color, respectively. The lower treatment performance was probably due to the increased TPOME concentrations. Based on the amounts of phenolic compounds, protein, and crude fiber in the grass biomass, the inoculated TPOME-treated grass had a satisfactory nutritional quality and digestibility for use as animal feed.

Biodegradation of crude oil using self-immobilized hydrocarbonoclastic deep sea bacterial consortium.

Hydrocarbonoclastic bacterial consortium that utilizes crude oil as carbon and energy source was isolated from marine sediment collected at a depth of 2100 m. Molecular characterization by 16S rRNA gene sequences confirmed that these isolates as Oceanobacillus sp., Nesiotobacter sp., Ruegeria sp., Photobacterium sp., Enterobacter sp., Haererehalobacter sp., Exiguobacterium sp., Acinetobacter sp. and Pseudoalteromonas sp. Self-immobilized consortium degraded more than 85% of total hydrocarbons after 10 days of incubation with 1% (v/v) of crude oil and 0.05% (v/v) of Tween 80 (non-ionic surfactant) at 28 ±â€¯2 °C. The addition of nitrogen and phosphorus sources separately i.e. 0.1% (v/v) of CO (NH2)2 or K2HPO4 enhanced the hydrocarbon utilization percentage. The pathways of microbial degradation of hydrocarbons were confirmed by FTIR, GC-MS, 1H and 13C NMR spectroscopy analyses. These results demonstrated a novel approach using hydrocarbonoclastic self-immobilized deep sea bacterial consortium for eco-friendly bioremediation.

Hydrocarbon degradation potential and competitive persistence of hydrocarbonoclastic bacterium Acinetobacter pittii strain ABC.

Acinetobacter pittii strain ABC was isolated from oily sludge sediments and characterized with regard to utilization/degradation of hydrocarbons and competitive persistence in hydrocarbon-amended media. The isolate grew in both aliphatic- and aromatic hydrocarbon-amended Bushnell-Haas medium (BHM). When incubated in 1% (v/v) Assam crude oil-amended BHM for 5 and 10 days, this strain was able to degrade 88% and 99.8% of the n-hexane extractable crude oil components, respectively. The isolate showed appreciable emulsification index (E24 65.26 ± 1.2%), hydrophobicity (60.88 ± 3.5%) and produced lipopeptide biosurfactant (0.57 g L-1). The isolate was able to tolerate heavy metal salts at concentrations reported in crude oil-polluted sediments from Assam. A 16S rDNA DGGE-based screening showed the persistence of A. pittii strain ABC in hydrocarbon-amended microcosms co-inoculated with other hydrocarbonoclastic bacterial strains (Pseudomonas aeruginosa AKS1, Bacillus sp. AKS2, Arthrobacter sp. BC1, and Novosphingobium panipatense P5:ABC), each isolated from the same oily sludge sediment. These findings indicate A. pittii strain ABC as a potential agent for the bioremediation of crude oil-polluted environment.

Crude Oil Degrading Fingerprint and the Overexpression of Oxidase and Invasive Genes for n-hexadecane and Crude Oil Degradation in the Acinetobacter pittii H9-3 Strain.

A crude oil-degrading bacterium named strain H9-3 was isolated from crude oil contaminated soil in the Northeastern area of China. Based on its morphological characteristics and 16S rDNA sequence analysis, strain H9-3 is affiliated to Acinetobacter pittii in the group of Gammaproteobacteria. The strain was efficient in removing 36.8% of the initial 10 g·L - 1 of crude oil within 21 days. GC-MS was performed and a preference was shown for n-C10, n-C11, i-C14, i-C17, i-C34, n-C12, n-C13, n-C14, n-C27, n-C32 and i-C13, over n-C16, n-C18⁻C22, n-C24⁻n-C31, and n-C36. This can be regarded as the specific fingerprint for crude oil degradation by strain H9-3 of Acinetobacter pittii. In addition to crude oil, it was shown that soybean oil and phenols can be utilized as carbon sources by strain H9-3. It was also shown that aniline and α -naphthol cannot be utilized for growth, but they can be tolerated by strain H9-3. Methylbenzene was neither utilized nor tolerated by strain H9-3. Although n-hexadecane was not preferentially consumed by strain H9-3, during culture with crude oil, it could be utilized for growth when it is the sole carbon source. The degradation of some branched alkanes (i-C14, i-C17 and i-C34) and the preferential degradation of crude oil over phenols could be used as a reference for distinguishing A. pittii from A. calcoaceticus. The difference in gene expression was very significant and was induced by diverse carbon sources, as shown in the qRT-PCR results. The oxidation and adhesion events occurred at high frequency during alkane degration by Acinetobacter pittii strain H9-3 cells.

Production, characterization, and applications of bioemulsifiers (BE) and biosurfactants (BS) produced by Acinetobacter spp.: A review.

Bioemulsifiers (BE) and biosurfactants (BS) are considered as multifunctional biomolecules of 21st century because of their functional abilities and eco-friendly properties. They are produced by various microorganisms under versatile and extreme environmental conditions. They have tremendous applications in various industries such as petroleum, food, medicine, pharmaceutical, chemical, paper & pulp, textile, and cosmetics. Currently, they are also considered as "green molecules" because of their wide applications in bioremediation of soil. Their importance has been increasing day by day in the global market as they are the natural resources with high-aggregate value. Although, there are numerous reports on BE and BS production by different bacteria, Acinetobacter spp. acquired special attention among all. This is because it is the earliest member known for the production of bioemulsifier. Emulsan and Alasan are the best examples of the commercially used BE produced by Acinetobacter spp. These BE are mainly used in microbial enhanced oil recovery and biodegradation of toxic compounds. This review is focused on BE and BS produced by Acinetobacter spp., their characterization and applications in different fields. This is the first review on genus Acinetobacter which defines independently about different types of BE and BS produced by it. It will also address the need of exploration of these molecules from various sources and their applications for the benefit of mankind and sustainable environment.

Characterization of Hydrocarbon-Degrading Bacteria in Constructed Wetland Microcosms Used to Treat Crude Oil Polluted Water.

Ten plant species were grown in constructed wetlands (CWs) to remediate water containing 2% (w/v) crude oil. The plant species with better growth and biomass production were Typha latifolia and Cyperus laevigatus, and they were significantly correlated (R2 = 0.91) with hydrocarbon degradation. From T. latifolia and C. laevigatus, 33 hydrocarbon-degrading bacterial strains were isolated from the rhizosphere, and root and shoot interiors. More diversified bacteria were found in the rhizosphere and endosphere of C. laevigatus than those of T. latifolia. The predominant cultural hydrocarbon-degrading bacteria were shown to belong to the genera Pseudomonas, Acinetobacter and Bacillus. In addition to genes involved in hydrocarbon degradation, most of the bacteria displayed multiple plant growth promoting (PGP) activities. This study suggests the importance of selecting suitable bacterial strains with hydrocarbon degradation and PGP activities for improving the efficacy of CWs used in remediating water contaminated with crude oil.

Draft genome Sequence of Phosphate-Accumulating Bacterium Acinetobacter tandoii SC36 from a Mangrove Wetland Ecosystem Provides Insights into Elements of Phosphorus Removal.

Acinetobacter tandoii SC36 was isolated from a mangrove wetland ecosystem in the Dongzhaigang Nature Reserve in Haikou, China. This bacterium was found to have a capacity for polyphosphate accumulation. To provide insight into its phosphorus metabolism and facilitate its application in phosphorus removal, we developed a draft genome of this strain. KEGG (Kyoto Encyclopedia of Genes and Genomes) annotation revealed three ppk genes and several phosphate metabolic related pathways in the genome of SC36. These genome data of Acinetobacter tandoii SC36 will facilitate elucidation of the mechanism of polyphosphate accumulation.

Overexpression of AdeABC efflux pump associated with tigecycline resistance in clinical Acinetobacter nosocomialis isolates.

OBJECTIVES: Tigecycline non-susceptible Acinetobacter nosocomialis (TNAN) has been discovered in clinical isolates. The resistance-nodulation-cell division (RND)-type efflux system plays a major role in tigecycline non-susceptible Acinetobacter baumannii, but the mechanism in A. nosocomialis remains unknown. Our aim was to analyse the contribution of efflux-based tigecycline resistance in clinical A. nosocomialis isolates collected from multiple medical centres in Taiwan. METHODS: A total of 57 A. nosocomialis isolates, including 46 TNAN and 11 tigecycline-susceptible A. nosocomialis (TSAN) isolates, were analysed. Of these, 46 TNAN isolates were clustered to ST410 (43 isolates) and ST68 (three isolates) by multi-locus sequence typing. RESULTS: The relationship between the RND efflux pump and tigecycline resistance was indirectly verified by successfully reducing tigecycline resistance with NMP, an efflux pump inhibitor. The three RND efflux systems (AdeABC, AdeIJK and AdeFGH) were detected in all clinical isolates. The transcript level of adeB gene increased significantly and was correlated with tigecycline resistance. Moreover, the AdeRS two-component system was further classified into four different types of AdeRS patterns considering the amino acid sequence. Further analysis showed that tigecycline resistance was related to the transcript level of adeB gene and the AdeRS pattern. CONCLUSION: This study showed that the dissemination of TNAN isolates in Taiwan is attributable mainly to the spread of ST410. The AdeABC efflux pump appeared to play an important role in the tigecycline resistance of A. nosocomialis.

Aerobic granular sludge and naphthenic acids treatment by varying initial concentrations and supplemental carbon concentrations.

Aerobic granular sludge (AGS) has previously been utilized in the treatment of toxic compounds due to its diverse and dense microbial structure. The present study subjected mature AGS to model naphthenic acids (NAs) representative of the Canadian oil sands. To this effect, three NA concentrations (10, 50 and 100 mg/L) and three supplemental carbon source concentrations (600, 1200 and 2500 mg/L) were studied in batch reactors for 5 days. The responding variables were chemical oxygen demand (COD), NA concentrations and nutrients. Cyclohexane carboxylic acid (CHCA), cyclohexane acetic acid (CHAA) and 1-adamantane carboxylic acid (ACA) were chosen to study structure-based degradation kinetics. The optimal COD according to the runs was 1200 mg/L. CHCA was removed completely with biodegradation rate constants increasing with lower NA concentrations and lower COD concentrations. CHAA was also removed completely, however, an optimal rate constant of 1.9 d-1 was achieved at NA and COD concentrations of 50 mg/L and 1200 mg/L, respectively. ACA removal trends did not follow statistically significant regressions; however, degradation and sorption helped remove ACA up to 19.9%. Pseudomonas, Acinetobacter, Hyphomonas and Brevundimonas spp. increased over time, indicating increased AGS adaptability to NAs.

Improved fatty aldehyde and wax ester production by overexpression of fatty acyl-CoA reductases.

BACKGROUND: Fatty aldehydes are industrially relevant compounds, which also represent a common metabolic intermediate in the microbial synthesis of various oleochemicals, including alkanes, fatty alcohols and wax esters. The key enzymes in biological fatty aldehyde production are the fatty acyl-CoA/ACP reductases (FARs) which reduce the activated acyl molecules to fatty aldehydes. Due to the disparity of FARs, identification and in vivo characterization of reductases with different properties are needed for the construction of tailored synthetic pathways for the production of various compounds. RESULTS: Fatty aldehyde production in Acinetobacter baylyi ADP1 was increased by the overexpression of three different FARs: a native A. baylyi FAR Acr1, a cyanobacterial Aar, and a putative, previously uncharacterized dehydrogenase (Ramo) from Nevskia ramosa. The fatty aldehyde production was followed in real-time inside the cells with a luminescence-based tool, and the highest aldehyde production was achieved with Aar. The fate of the overproduced fatty aldehydes was studied by measuring the production of wax esters by a native downstream pathway of A. baylyi, for which fatty aldehyde is a specific intermediate. The wax ester production was improved with the overexpression of Acr1 or Ramo compared to the wild type A. baylyi by more than two-fold, whereas the expression of Aar led to only subtle wax ester production. The overexpression of FARs did not affect the length of the acyl chains of the wax esters. CONCLUSIONS: The fatty aldehyde production, as well as the wax ester production of A. baylyi, was improved with the overexpression of a key enzyme in the pathway. The wax ester titer (0.45 g/l) achieved with the overexpression of Acr1 is the highest reported without hydrocarbon supplementation to the culture. The contrasting behavior of the different reductases highlight the significance of in vivo characterization of enzymes and emphasizes the possibilities provided by the diversity of FARs for pathway and product modulation.

Effect of a static magnetic field on the microscopic characteristics of highly efficient oil-removing bacteria.

To better understand the microbial oil removal enhancement process by a magnetic field, the effect of a static magnetic field (SMF) on the microscopic characteristics of highly efficient biodegradation oil-removing bacteria was studied. The Acinetobacter sp. B11 strain with a 53.6% oil removal rate was selected as the reference bacteria. The changes in the microscopic characteristics of Acinetobacter sp. B11 such as the cell surface morphology, cell permeability and cell activity of the bacteria were investigated. The results showed that low-intensity magnetic fields (15-35 mT) improved the ability of Acinetobacter sp. B11 to remove oil by 11.9% at 25 mT compared with that of bacteria with no magnetic field. Without destroying the cell membrane, the low-intensity magnetic fields increased the cell membrane permeability and improved the activity of superoxide dismutase (SOD), which effectively enhanced the oil degradation performance of the bacteria.

Piggery wastewater treatment by Acinetobacter sp. TX5 immobilized with spent mushroom substrate in a fixed-bed reactor.

Acinetobacter sp. TX5 immobilized with spent Hypsizygus marmoreus substrate (SHMS) was used to treat the raw piggery wastewater (RPW). In batch experiments, NH4+-N in the diluted RPW decreased from initial 34.95 mg/L to 3.83 mg/L at 8 h with the removal efficiency (RE) being 89%, and the beads immobilized with SHMS were comparable to those immobilized with activated carbon. In continuous experiments, the RE ranged from 74% to 95% for NH4+-N, from 73% to 93% for TN and from 54% to 82% for COD when the RPW was treated in a fixed-bed reactor packed with SHMS-immobilized TX5. The isotope analysis and enzyme purification indicated simultaneous nitrification and denitrification existing in TX5. This is the first time that spent mushroom substrates have been used to immobilize Acinetobacter species to treat the real RPW and a denitrifying nitrite reductase (dNiR) has been purified to make the nitrogen removal pathway in this species clearer.

Ni(II) and Cu(II) removal from aqueous solution by a heavy metal-resistance bacterium: kinetic, isotherm and mechanism studies.

The potentiality of a heavy metal-resistance bacterium Acinetobacter sp. HK-1 for removing Ni(II) and Cu(II) ions from aqueous solution and the biosorption mechanism were investigated in this study. The effects of pH, contact time and Ni(II)/Cu(II) concentration on the adsorption process were evaluated and the maximum biosorption capacity of strain HK-1 was found to be 56.65 mg/g for Ni(II) and 157.2 mg/g for Cu(II), respectively. The experimental kinetic data fit well with the pseudo-second-order model (R2 > 0.98) and the biosorption process was best explained by the Langmuir-Freundlich dual model (R2 > 0.97). The morphologies of HK-1 before and after adsorption in a Ni(II)/Cu(II) supplemented system were compared using a scanning electron microscope. After adsorption, the valence state of Ni(II)/Cu(II) was not changed and the formation of nickel/copper phosphate was observed using X-ray photoelectron spectroscopy (XPS) and X-ray diffraction. The results of Fourier transform infrared spectroscopy and XPS further indicated that amine, phosphate and carboxyl groups were involved in the biosorption process. Cu(II) biosorption by Acinetobacter sp. was firstly reported. Based on the above results, it can be concluded that Acinetobacter sp. HK-1 has a promising application in Ni(II) and Cu(II) ion removal from industrial wastewater.

Phosphorus removal performance and population structure of phosphorus-accumulating organisms in HA-A/A-MCO sludge reduction process.

We developed a new sludge reduction HA-A/A-MCO (Hydrolysis-Acidogenosis-Anaerobic/Anoxic -Multistep Continuous Oxic tank) process, which has improved phosphate (P) and nitrogen (N) removal. Its biological treatment unit uses an A2/O P & N removal process with hydrolysis acidification, multistep continuous aeration, and continuous flow, coupled with sidestream P removal by draining out anaerobic P-bearing wastewater. The process has advanced synchronization of P and N removal and sludge reduction. The improved performance is closely associated with the population structure of P-accumulating organisms (PAOs). This study investigated the relationship between P removal performance and the population structure of PAOs. The results show that the average effluent P content of HA-A/A-MCO process was only 0.44 mg/L, when the influent P concentration was 8∼12 mg/L. The effluent met the A standard set by GB18918-2002. PAOs were able to effectively release 1 mg of P and absorb 2.8 mg of P. The system removed P by draining out anaerobic P-rich wastewater, as P had been reduced in the aerobic absorption process. This reduced the need for excess P uptake ability of the PAOs. The bacterial pure culture method was applied to isolate 5 PAOs with typical P absorption and removel features. 16SrDNA amplification and sequence analysis revealed that Acinetobacter sp. and Lampropedia sp played dominant roles in anaerobic P-releasing process. Moreover, Devosia sp. and Bdellovibrio sp were the primary strains in the aerobic tank, and, they were the major stains for P absorption. Uncultured Bacterium and other uncultured strains were detected in the anoxic tank.

Genomic and phenotypic characterization of the species Acinetobacter venetianus.

Crude oil is a complex mixture of hydrocarbons and other organic compounds that can produce serious environmental problems and whose removal is highly demanding in terms of human and technological resources. The potential use of microbes as bioremediation agents is one of the most promising fields in this area. Members of the species Acinetobacter venetianus have been previously characterized for their capability to degrade n-alkanes and thus may represent interesting model systems to implement this process. Although a preliminary experimental characterization of the overall hydrocarbon degradation capability has been performed for five of them, to date, the genetic/genomic features underlying such molecular processes have not been identified. Here we have integrated genomic and phenotypic information for six A. venetianus strains, i.e. VE-C3, RAG-1(T), LUH 13518, LUH 7437, LUH 5627 and LUH 8758. Besides providing a thorough description of the A. venetianus species, these data were exploited to infer the genetic features (presence/absence patterns of genes) and the short-term evolutionary events possibly responsible for the variability in n-alkane degradation efficiency of these strains, including the mechanisms of interaction with the fuel droplet and the subsequent catabolism of this pollutant.