Denitrification using PBS as carbon source and biofilm support in a packed-bed bioreactor

Biodegradable polymer was used as carbon source and biofilm support for nitrate removal from aqueous solution as an attractive alternative for biological denitrification. The objective of this paper was to investigate the denitrification performance and microbial community of a packed-bed bioreactor using poly (butanediol succinate) (PBS), a biodegradable polymer, as carbon source and biofilm support. NO₃–N concentration was determined by UV spectrophotometer. NO₂–N concentration was assayed by hydrochloric acid naphthyl ethylenediamine spectrophotometry method. Total organic carbon (TOC) was measured using a TOC analyzer. The morphology of the samples was observed using an environmental scanning electron microscope (ESEM). The microbial community was analyzed by pyrosequencing method. The experimental results showed that an average removal efficiency of nitrate was 95 %. ESEM observation and FTIR analysis indicated the changes of PBS granules before and after microbial utilization. Pyrosequencing results showed that Betaproteobacteria predominated, and most of PBS-degrading denitrifying bacteria were assigned to the family Comamonadaceae. Denitrifying bacteria accounted for 13.02 % in total population. The PBS granules were suitable support and carbon source for denitrifying bacteria.     

Biofilm comprising phototrophic, diazotrophic, and hydrocarbon-utilizing bacteria: a promising consortium in the bioremediation of aquatic hydrocarbon pollutants

Biofilms harboring simultaneously anoxygenic and oxygenic phototrophic bacteria, diazotrophic bacteria, and hydrocarbon-utilizing bacteria were established on glass slides suspended in pristine and oily seawater. Via denaturing gradient gel electrophoresis analysis on PCR-amplified rRNA gene sequence fragments from the extracted DNA from biofilms, followed by band amplification, biofilm composition was determined. The biofilms contained anoxygenic phototrophs belonging to alphaproteobacteria; pico- and filamentous cyanobacteria (oxygenic phototrophs); two species of the diazotroph Azospirillum; and two hydrocarbon-utilizing gammaproteobacterial genera, Cycloclasticus and Oleibacter. The coexistence of all these microbial taxa with different physiologies in the biofilm makes the whole community nutritionally self-sufficient and adequately aerated, a condition quite suitable for the microbial biodegradation of aquatic pollutant hydrocarbons.     

混合型污水处理厂原位生物强化脱氮中试及微生物群落结构分析研究

针对混合型污水处理厂进水水质波动大、C/N低、总氮稳定达标困难等共性问题,开发出适宜于低C/N、贫营养等苛刻环境条件的高效脱氮菌剂.现场中试结果表明:经过脱氮菌强化后,生化系统脱氮效率提升8.5百分点,出水平均总氮较强化前下降3 mg/L,出水水质稳定性明显提升,生物强化效果显著.在不补加碳源条件下,深度处理出水总氮完...     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

Investigation of solid-phase buffers for sulfur-oxidizing autotrophic denitrification.

This paper investigates biological denitrification using autotrophic microorganisms that use elemental sulfur as an electron donor. In this process, for each gram of nitrate-nitrogen removed, approximately 4.5 g of alkalinity (as calcium carbonate) are consumed. Because denitrification is severely inhibited below pH 5.5, and alkalinity present in the influent wastewaters is less than the alkalinity consumed, an external buffer was needed to arrest any drop in pH from alkalinity consumption. A packed-bed bioreactor configuration is ideally suited to handle variations in flow and nitrate loading from decentralized wastewater treatment systems, as it is a passive system and thus requires minimal maintenance; therefore, a solid-phase buffer packed with the elemental sulfur in the bioreactor is most suitable. In this research, marble chips, limestone, and crushed oyster shells were tested as solid-phase buffers. Bench- and field-scale studies indicated that crushed oyster shell was the most suitable buffer based on (1) the rate of dissolution of buffer and the buffering agent released (carbonate, bicarbonate, or hydroxide), (2) the ability of the buffer surface to act as host for microbial attachment, (3) turbidity of the solution upon release of the buffering agent, and (4) economics.     

Microbiological analysis of multi-level borehole samples from a contaminated groundwater system

A range of bacteriological, geochemical process-related and molecular techniques have been used to assess the microbial biodegradative potential in groundwater contaminated with phenol and other tar acids. The contaminant plume has travelled 500 m from the pollutant source over several decades. Samples were obtained from the plume using a multi-level sampler (MLS) positioned in two boreholes (boreholes 59 and 60) which vertically transected two areas of the plume. Activity of the microbial community, as represented by phenol degradation potential and ability to utilise a range of substrates, was found to be influenced by the plume. Phenol degradation potential appeared to be influenced more by the concentration of the contaminants than the total bacterial cell numbers. However, in the areas of highest phenol concentration, the depression of cell numbers clearly had an effect. The types of bacteria present were assessed by culture and DNA amplification by polymerase chain reaction (PCR). Bacterial groups or processes associated with major geochemical processes, such as methanogenesis, sulphate reduction and denitrification, that have the potential to drive contaminant degradation, were detected at various borehole levels. A comparative molecular analysis of the microbial community between samples obtained from the MLS revealed the microbial community was diverse. The examination of microbial activity complemented those results obtained through chemical analysis, and when combined with hydrological data, showed that MLS samples provided a realistic profile of plume effects and could be related to the potential for natural attenuation of the site.     

光能自养生物膜形成机制及脱氮影响因素的研究进展

光能自养生物膜是一个由复杂微生物群落构成的相对稳定的微生态系统,微生物细胞及非生物物质镶嵌在微生物分泌的有机聚合物基质上,其显著特征是膜表面寄居着光能自养生长的微生物,并能在碳源缺乏条件下完成生物脱氮.总结了光能自养生物膜的形成机制,概述了光照、温度、微生物种类、营养物浓度对光能自养生物膜脱氮的影响,指出光能自养生物膜...     

Nitrate removal from groundwater using biodegradable polymers as carbon source and biofilm support

A new kind of biodegradable polymer, PBS, was used as both carbon source and biofilm support in a fixed-bed reactor to remove nitrate from groundwater. The experimental results showed that the denitrifying bacteria can easily attach to the surface of PBS granules and adapt to use PBS as carbon source. The SEM observation indicated that the fine biofilm can develop on the surface of PBS granules within 15 days. The denitrification rate increased with temperature increase in the range of 10-35°C, the maximum denitrification rate reached 1.00 mg/g.d at 35°C. Continuous experiment results showed that nitrate (50 mg/l) can be removed effectively and nitrite did not accumulate in the effluent.     

生物流化床反应器生物膜特性研究进展

生物流化床技术应用于废水处理领域具有广阔的发展前景。生物流化床反应器对污染物的降解主要依赖于活性生物膜 ,研究生物膜的特性是提高反应器效率的重要手段。本文对描述生物膜特性的主要参数进行了综述 ,其中 ,生物膜种群构成的差异是影响反应器效率的最重要因素 ,本文重点探讨了生物膜中微生物的种群分布特征、种群的生态演替过程与反应器性能的关系。在此基础上 ,提出了对生物流化床生物膜特性进行强化的几种手段     

Biofilm formation and microbial community analysis of the simulated river bioreactor for contaminated source water remediation

Background, aim, and scope

The start-up pattern of biofilm remediation system affects the biofilm characteristics and operating performances. The objective of this study was to evaluate the performances of the contaminated source water remediation systems with different start-up patterns in view of the pollutants removal performances and microbial community succession.

Methods

The operating performances of four lab-scale simulated river biofilm reactors were examined which employed different start-up methods (natural enrichment and artificial enhancement viadischarging sediment with influent velocity gradient increase) and different bio-fillers (Elastic filler and AquaMats? ecobase). At the same time, the microbial communities of the bioreactors in different phases were analyzed by polymerase chain reaction, denaturing gradient gel electrophoresis, and sequencing.

Results and discussion

The pollutants removal performances became stable in the four reactors after 2 months?? operation, with ammonia nitrogen and permanganate index (COD_Mn) removal efficiencies of 84.41?C94.21% and 69.66?C76.60%, respectively. The biomass of mature biofilm was higher in the bioreactors by artificial enhancement than that by natural enrichment. Microbial community analysis indicated that elastic filler could enrich mature biofilm faster than AquaMats?. The heterotrophic bacteria diversity of biofilm decreased by artificial enhancement, which favored the ammonia-oxidizing bacteria (AOB) developing on the bio-fillers. Furthermore, Nitrosomonas- and Nitrosospira-like AOB coexisted in the biofilm, and Pseudomonas sp., Sphaerotilus sp., Janthinobacterium sp., Corynebacterium aurimucosum were dominant in the oligotrophic niche.

Conclusion

Artificial enhancement via the combination of sediment discharging and influent velocity gradient increasing could enhance the biofilm formation and autotrophic AOB enrichment in oligotrophic niche.     

硫自养反硝化去除地下水中硝酸盐氮的研究

研究实际地下水硫自养反硝化动力学过程，考察季节因素(温度)对动力学的影响，实验结果表明，地下水升流式硫自养滤柱反硝化动力学符合1/2级动力学模型，其反应速率常数受温度的影响很大，用阿仑尼乌斯方程计算硫自养反硝化活化能为80.38 kJ/mol。硫自养反硝化产生的硫酸根与反硝化掉的硝酸根离子呈线性相关。在地下水不经任何预处理的条件下，硫自养反硝化仍能有效地脱除地下水中的硝酸盐，反应器出水的pH值仍维持在中性范围。     

Antimicrobial potential of the ionophore monensin on freshwater biofilm bacteria

Microorganisms play key roles in stream ecosystems, but comparatively little is known about the resilience of freshwater bacterial communities and their susceptibility to the chemical by-products of agricultural land use. Antibiotics used in the agricultural sector are of particular concern and have been detected in waterways associated with agricultural land. Despite widespread agricultural intensification globally and the sector's high antibiotic use, the effects of agricultural antibiotic by-products on stream microbial communities have yet to be characterised. We investigated the impacts of the antibiotic monensin on microbial biofilm communities in a simulated contamination event using streamside-replicated channels. A 24-h pulse experiment in flow channels precolonised by stream biofilm microbial communities contrasted the effects of monensin concentrations ranging from realistic to extreme toxicity levels (1–550 ug L^?1). Biofilm community composition was characterised immediately before and after the pulse for several weeks using automated ribosomal intergenic spacer analysis. Despite applying acutely toxic levels of monensin, only limited effects to biofilm community composition were detected immediately after antibiotic application, and these disappeared within 4 days. Rather, temporal factors drove biofilm differences, highlighting the overriding importance of wider, catchment-level, physiochemical hydrological influences on structuring freshwater biofilm communities, as opposed to localised and sporadic agricultural surface runoff contamination events containing antibiotics.     

同步硝化反硝化好氧颗粒污泥微生物相和氮去除能力的研究

对颗粒污泥中的微生物菌群进行了分离和鉴定，并确定纯种微生物的脱氮能力。颗粒污泥中含假单胞菌属5株。亚硝化单胞菌属7株，硝化杆菌属4株，气单胞菌属3株，黄单胞菌属2株，孢杆菌属4株，黄色杆菌属2株和产碱菌属2株。这些菌属中包含了参与硝化反应的亚硝化单胞菌属和硝化杆菌属微生物，以及有好氧反硝化能力的Pseudomonas和Alcaligenes菌。29株微生物摇瓶混合培养的脱氮试验结果表明，30h内有机物去除率为75％，氮去除率近50％。     

降解SO_2功能菌的16S rRAN基因测序及降解特性

用低浓度SO2诱导驯化方法获得高效脱硫菌群,并用分离培养与16S rRNA基因测序技术相结合的方法鉴定菌群种属,分析驯化过程中种群结构的动态变化,同时研究分离纯菌种的脱硫性能。结果表明,从诱导驯化7 d和14 d菌液中分别分离出23株菌和22株菌,16S rRNA序列分析发现这些菌归属于13个种,其中有6个种（Rhodococcus erythropolis、Pseudomonas putida、Microbacterium oxydans、Sphingomonas koreensis、Acinetobacter junii、Acinetobacter johnsonii）对SO2-3有较强的降解能力,并在持续驯化过程中稳定的生长传代,降解产物以硫酸根为主,还有极少量的单质硫。与含混合菌的驯化菌液降解SO2-3的能力相比,单一脱硫菌的脱硫性能较弱。脱硫功能菌株及其基本特性的研究为微生物处理SO2烟气提供了丰富的菌源信息和理论基础。     

Salt marsh plants as key mediators on the level of cadmium impact on microbial denitrification

The fate of excess nitrogen in estuaries is determined by the microbial-driven nitrogen cycle, being denitrification a key process since it definitely removes fixed nitrogen as N₂. However, estuaries receive and retain metals, which may negatively affect this process efficiency. In this study, we evaluated the role of salt marsh plants in mediating cadmium (Cd) impact on microbial denitrification process. Juncus maritimus and Phragmites australis from an estuary were collected together with the sediment involving their roots, each placed in vessels and maintained in a greenhouse, exposed to natural light, with tides simulation. Similar non-vegetated sediment vessels were prepared. After 3 weeks of accommodation, nine vessels (three per plant species plus three non-vegetated) were doped with 20 mg/L Cd²⁺ saline solution, nine vessels were doped with 2 mg/L Cd²⁺ saline solution and nine vessels were left undoped. After 10 weeks, vessels were dissembled and denitrification potential was measured in sediment slurries. Results revealed that the addition of Cd did not cause an effect on the denitrification process in non-vegetated sediment but had a clear stimulation in colonized ones (39 % for P. australis and 36 % for J. maritimus). In addition, this increase on denitrification rates was followed by a decrease on N₂O emissions and on N₂O/N₂ ratios in both J. maritimus and P. australis sediments, increasing the efficiency of the N₂O step of denitrification pathway. Therefore, our results suggested that the presence of salt marsh plants functioned as key mediators on the degree of Cd impact on microbial denitrification.     

Biological denitrification of high strength nitrate waste using preadapted denitrifying sludge

Denitrification of synthetic high nitrate waste containing 9032 ppm NO(3)-N (40,000 ppm NO(3)) in a time period of only 6h has been achieved in our previous study using activated sludge. The activated sludge culture was acclimatized by a stepwise increase in the nitrate concentration of synthetic waste. In the present work, studies were carried out on the changing microbial population of the sludge and the physiology of nitrate metabolism during the various stages of adaptation process to high strength synthetic nitrate waste. During the course of adaptation, with an increase in the nitrate concentration, a sharp increase in the number of denitrifiers was found with an equally rapid decrease in the nitrifying community. Two key enzymes involved in the first two steps of the denitrification process were also studied during this period. The results of the study suggest that specific enzyme levels increase as the activated sludge adapts itself to higher nitrate concentrations. Biological denitrification of high nitrate waste is a slow process and to increase the rate of denitrification, parameters such as pH, temperature, C:N and biomass concentration of the process were optimized using orthogonal array method. Optimized conditions increased the specific nitrate reduction rate by 54% and specific nitrite reduction rate by 45%.     

Biodegradation rates of 2-methylisoborneol (MIB) and geosmin through sand filters and in bioreactors

Taste and odour (T&O) causing compounds, in particular, 2-methylisoborneol (MIB) and geosmin, are a problem for water authorities as they are recalcitrant to conventional water treatment. In this study, biological sand filtration was shown to be an effective process for the complete removal of MIB and geosmin, with removal shown to be predominantly through biodegradation. In addition, MIB and geosmin were also effectively degraded in batch bioreactor experiments using biofilm sourced from one of the sand filters as the microbial inoculum. The biodegradation of MIB and geosmin was determined to be a pseudo-first-order reaction with rate constants ranging between 0.10 and 0.58 d⁻¹ in the bioreactor experiments. Rate constants were shown to be dependent upon the initial concentration of the microbial inoculum but not the initial concentration of MIB and geosmin when target concentrations of 200 and 50 ng l⁻¹ were used. Furthermore, rate constants were shown to increase upon re-exposure of the biofilm to both T&O compounds. Enrichment cultures with subsequent community profile analysis using 16S rRNA-directed PCR-DGGE identified four bacteria most likely involved in the biodegradation of geosmin within the sand filters and bioreactors. These included a Pseudomonas sp., Alphaproteobacterium, Sphingomonas sp. and an Acidobacteriaceae member.     

The performance of an anaerobic sequencing batch biofilm reactor treating domestic sewage colonized by anoxygenic phototrophic bacteria

There are few reports on morphological characterization of microbial population colonizing anaerobic bioreactors and the aim of this work was to access such variable in an anaerobic sequencing batch biofilm reactor treating the University of Sao Paulo (Sao Carlos city, Brazil) domestic sewage. This pilot-scale reactor (1.2m3) has been treating 0.65 m3 of liquid waste under cycles of 8h. The ASBBR has the distinct characteristics of being filled with support material for biomass attachment with the aim of skipping the sedimentation phase during the operational cycles, as it is commonly observed in anaerobic sequencing batch reactors (ASBR). Physical, chemical and physico-chemical variables were accessed in the influent and in the effluent for performance evaluation. Microbial characterization was made by means of direct microscopy and samples were taken over 150 d with a 25 d period interval. The ASBBR attained approximately 60% of COD removal efficiency. Microscopic analysis of biomass showed the presence of anoxygenic phototrophic bacteria probably influencing the ASBBR performance in the domestic sewage treatment. It is very likely that the exclusion of phototrophic sulfur bacteria by efficiently restraining the light would enhance the bioreactor efficiency.     

Nitrifying community analysis in a single submerged attached-growth bioreactor for treatment of high-ammonia waste stream.

This study investigated the nitrifying community structure in a single-stage submerged attached-growth bioreactor (SAGB) that successfully achieved stable nitrogen removal over nitrite of a high-strength ammonia wastewater. The reactor was operated with intermittent aeration and external carbon addition (methanol). With influent ammonia and total Kjeldahl nitrogen ranging from 537 to 968 mg/L and 643 to 1510 mg/L, respectively, 85% nitrogen removal was obtained, and effluent was dominated by nitrite (NO2-/NOx > 0.95). Nitrifying community analysis using fluorescence in situ hybridization (FISH), with a hierarchical set of probes targeting known ammonia-oxidizing bacteria (AOB) within beta-proteobacteria, showed that the AOB community of the biofilter consists almost entirely of members of the Nitrosomonas europaea/eutropha and the Nitrosococcus mobilis lineages. Image analysis of FISH pictures was used to quantify the identified AOB, and it was estimated that Nitrosomonas europaea/eutropha-like AOB accounted for 4.3% of the total volume of the biofilm, while Nitrosococcus mobilis-like AOB made up 1.2%; these numbers summed up to a total AOB fraction of 5.5% of the total volume on the biofilm. Nitrite-oxidizing bacteria (NOB) were not detectable in the biofilm samples with probes for either Nitrospira sp. or Nitrobacter sp., which indicated that NOB were either absent from the biofilters or present in numbers below the detection limit of FISH (< 0.1% of the total biofilm). Nitrite oxidizers were likely outcompeted from the system because of the free ammonia inhibition and the possibility that the aeration period (from intermittent aeration) was not sufficiently long for the NOB to be released from the competition for oxygen with heterotrophs and AOB. The nitrogen removal via nitrite in a SAGB reactor described in this study is applicable for high-ammonia-strength wastewater treatment, such as centrate or industrial wastes.