Microbial fouling community analysis of the cooling water system of a nuclear test reactor with emphasis on sulphate reducing bacteria

Culture and molecular-based techniques were used to characterize bacterial diversity in the cooling water system of a fast breeder test reactor (FBTR). Techniques were selected for special emphasis on sulphate-reducing bacteria (SRB). Water samples from different locations of the FBTR cooling water system, in addition to biofilm scrapings from carbon steel coupons and a control SRB sample were characterized. Whole genome extraction of the water samples and SRB diversity by group specific primers were analysed using nested PCR and denaturing gradient gel electrophoresis (DGGE). The results of the bacterial assay in the cooling water showed that the total culturable bacteria (TCB) ranged from 10³ to 10⁵ cfu ml^?1; iron-reducing bacteria, 10³ to 10⁵ cfu ml^?1; iron oxidizing bacteria, 10² to 10³ cfu ml^?1 and SRB, 2–29 cfu ml^?1. However, the counts of the various bacterial types in the biofilm sample were 2–3 orders of magnitude higher. SRB diversity by the nested PCR-DGGE approach showed the presence of groups 1, 5 and 6 in the FBTR cooling water system; however, groups 2, 3 and 4 were not detected. The study demonstrated that the PCR protocol influenced the results of the diversity analysis. The paper further discusses the microbiota of the cooling water system and its relevance in biofouling.     

Biofouling and microbial corrosion problem in the thermo-fluid heat exchanger and cooling water system of a nuclear test reactor

This article discusses aspects of biofouling and corrosion in the thermo-fluid heat exchanger (TFHX) and in the cooling water system of a nuclear test reactor. During inspection, it was observed that >90% of the TFHX tube bundle was clogged with thick fouling deposits. Both X-ray diffraction and Mössbauer analyses of the fouling deposit demonstrated iron corrosion products. The exterior of the tubercle showed the presence of a calcium and magnesium carbonate mixture along with iron oxides. Raman spectroscopy analysis confirmed the presence of calcium carbonate scale in the calcite phase. The interior of the tubercle contained significant iron sulphide, magnetite and iron-oxy-hydroxide. A microbiological assay showed a considerable population of iron oxidizing bacteria and sulphate reducing bacteria (10⁵ to 10⁶ cfu g^?1 of deposit). As the temperature of the TFHX is in the range of 45–50°C, the microbiota isolated/assayed from the fouling deposit are designated as thermo-tolerant bacteria. The mean corrosion rate of the CS coupons exposed online was ～2.0 mpy and the microbial counts of various corrosion causing bacteria were in the range 10³ to 10⁵ cfu ml^?1 in the cooling water and 10⁶ to 10⁸ cfu ml^?1 in the biofilm.     

Microbial community analysis of rectal methanogens and sulfate reducing bacteria in two non-human primate species

Background  Methanogenesis by methanogenic Archaea and sulfate reduction by sulfate reducing bacteria (SRB) are the major hydrogenotrophic pathways in the human colon. Methanogenic status of mammals is suggested to be under evolutionary rather than dietary control. However, information is lacking regarding the dynamics of hydrogenotrophic microbial communities among different primate species.
Methods  Rectal swabs were collected from 10 sooty mangabeys ( Cercocebus atys ) and 10 baboons ( Papio hamadryas ). The diversity and abundance of methanogens and SRB were examined using PCR-denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR (qPCR).
Results  The DGGE results revealed that intestinal Archaea and SRB communities differ between mangabeys and baboons. Phylogenetic analyses of Archaea DGGE bands revealed two distinct clusters with one representing a putative novel order of methanogenic Archaea. The qPCR detected a similar abundance of methanogens and SRB.
Conclusions  Intestinal Archaea and SRB coexist in these primates, and the community patterns are host species-specific.     

利用光合细菌进行微生物修复：一种降低辛硫磷在养殖水中积累的低成本方法

【背景】中国是农业生产大国,渔林农牧占比庞大。有机农药无论在畜牧业还是水产养殖业都有广泛的应用。有机磷农药(Organophosphorus Pesticide,OP)是应用最广泛的有机农药,具有低毒和不易残留的优点。OP在水体中大量积累可通过生物富集作用间接影响人体健康,由此产生的生殖毒性不容忽视。光合细菌作为环境友好型水体有益菌,部分菌种具有降解有机农药的功能。【目的】自上海海洋大学明湖中分离纯化得到一株光合细菌(编号SPZ)。探究其对辛硫磷的耐受程度及降解效果,为养殖水体中有机磷农药的生物降解提供目的菌株。【方法】利用16S rRNA基因序列分析方法对目标菌株进行种属鉴定;利用紫外分光光度法测定分离菌株SPZ和标准菌株ST在不同接种量下的OD660并测定实验周期内光合细菌在不同浓度辛硫磷中OD660的变化趋势,以示辛硫磷对光合细菌的毒性作用;利用高效液相色谱法(High Performance Liquid Chromatography,HPLC)测定菌株对水体辛硫磷的降解能力;通过HPLC测定加热致死菌与活菌对水体辛硫磷的降解能力,确定菌株对辛硫磷的降解方式。【结果】16S r...     

Microbial biomass and community structure of a phase-separated methanogenic reactor determined by lipid analysis

Summary An anaerobic phase-separation biomass reactor was established on cellulose with the hydrolysis and fermentation steps occurring in the first stage, and acetogenesis and methanogenesis in the second stage. Based upon lipid biomarker analysis, eubacterial and eukaryotic cells accounted for approximately 6% of the volatile solids of the first stage and 17% of the second, while methanogens were approximately 1% of the volatile solids in the first stage and 9% of the second. Clustering the polar lipid fatty acids into groups based upon their distributions between the two stages of the reactor clarified the differences in community structure caused by phase-separated operation. Although inoculated from the same source, the two stages maintained very different microbial communities. Signature fatty acids known as indicators of unbalanced growth in eubacteria were significantly higher in the first stage of the reactor.     

Microbial community analysis of a biogas-producing completely stirred tank reactor fed continuously with fodder beet silage as mono-substrate

The bioconversion of renewable raw material to biogas by anaerobic microbial fermentation processes in completely stirred tank reactors (CSTR) is a valuable alternative resource of energy especially for rural areas. However, knowledge about the microorganisms involved in the degradation of plant biomass is still poor. In this study, a first analysis of the biogas-forming process within a CSTR fed continuously with fodder beet silage as mono-substrate is presented in the context of molecular data on the microbial community composition. As indicated by the conventional process parameters like pH value, content of volatile fatty acids, N:P ratio and the biogas yield, the biogas-forming process within the CSTR occurred with a stable and efficient performance. The average biogas yield based on volatile solids was 0.87m(3)kg(-1) at an organic loading rate of 1.2-2.3kgm(-3)d(-1). This amounts to 94% of the theoretical maximum. In order to identify microorganisms within the CSTR, a 16S rDNA clone library was constructed by PCR amplification applying a prokaryote-specific primer set. One hundred and forty seven clones were obtained and subsequently characterized by amplified rDNA restriction analysis (ARDRA). The sequences of 60 unique ARDRA patterns were estimated in a length of approximately 800-900bp each. Four of them were assigned to the domain Archaea and 56 to the domain Bacteria. Within the domain Archaea, all clones showed a close relationship to methanogenic species. Major bacterial groups represented in the clone library were the class Clostridia of the phylum Firmicutes (22% of all 16S rDNA clones), the class Deltaproteobacteria of the phylum Proteobacteria (24%), the class Bacilli of the phylum Firmicutes (22%) and members of the phylum Bacteroidetes (21%). Within these major groups, the highest biodiversity was found within the class Clostridia (35% of all operational taxonomic units). Members of the phyla Actinobacteria and Spirochaetes were represented only by 5 and 2 clonal sequences, respectively.     

Microbial community structure and operational performance of a fluidized bed biofilm reactor treating oil sands process-affected water

This study evaluated the treatment of oil sands process-affected water (OSPW) using a fluidized bed biofilm reactor (FBBR) with granular activated carbon (GAC) as support media. The bioreactor was operated for 120 days at different organic and hydraulic loading rates. The combined GAC adsorption and biodegradation process removed 51% of chemical oxygen demand (COD), 56% of acid-extractable fraction (AEF) and 96% of classical naphthenic acids (NAs) under optimized operational conditions. Bioreactor treatment efficiencies were dependent on the organic loading rate (OLR), and to a lower degree, on the hydraulic loading rate (HLR). Further ultra performance liquid chromatography/high resolution mass spectroscopy (UPLC/HRMS) analysis showed that the removal of classical NAs increased as the carbon number increased. Compared with planktonic bacterial community in OSPW, more diverse microbial structures were found in biofilms colonized on the surface of GAC after 120-day treatment, with various carbon degraders namely Polaromonas jejuensis, Algoriphagus sp., Chelatococcus sp. and Methylobacterium fujisawaense in the GAC-biofilm reactor. The results of this study, therefore, showed that the GAC-biofilm seems to be a promising biological treatment method for OSPW remediation.     

Microbial community structure analysis of produced water from a high-temperature North Sea oil-field

Molecular and culture-based methods were used to investigate the microbial diversity in produced water obtained from the high-temperature Troll oil formation in the North Sea. 16S rRNA gene libraries were generated from total community DNA, using universal archaeal or bacterial oligonucleotide primer sets. Sequence analysis of 88 clones in the bacterial library indicated that they originated from members of Firmicutes (48 sequences), Bacteroidetes (17 sequences), δ-Proteobacteria (15 sequences), Spirochaetes (5 sequences), Thermotogales (2 sequences) and γ-Proteobacteria (1 sequence). Twenty-two sequences in the archaeal library were close relatives to members of the genera Methanococcus (18 sequences), Methanolobus (3 sequences) and Thermococcus (1 sequence). Most of the bacterial sequences shared less than 95% identity with their closest match in GenBank, indicating that the produced water harbours a unique community of novel bacterial species or genera. Members of the thermophilic genera Thermosipho, Thermotoga, Anaerophaga and Thermovirga were isolated. The Troll formations are not injected with sea water. Thus, dramatic changes of the in situ conditions have been avoided, and a common source of continuous contamination from injection water can be excluded. However, the majority of the organisms detected in the gene libraries were most closely related to cultivated organisms with optimum temperatures for growth well below the in situ reservoir temperature (70°C), indicating that produced water from the Troll platform harbours a substantial amount of non-indigenous organisms. This was confirmed by the isolation of a number of mesophilic and moderately thermophilic organisms that were unable to grow at reservoir temperature.     

城市自然水体微生物种群分析及溶藻蛋白酶初步研究

采用PCR-DGGE技术,研究了城市自然水体中微生物菌群结构及投加不同浓度的铜绿微囊藻对水体微生物种群结构变化的影响,同时探讨了藻类诱导水体中微生物产生溶藻蛋白酶的作用;结果表明自然水体中微生物多样性十分丰富,加藻冲击后水体微生物种群结构没有明显变化,个别菌种的优势发生演替;微生物溶藻蛋白酶被诱导,通过片段序列分析,可以基本确定为微生物Clostridium botulinum体内溶藻酶.     

Microbial community profile of a lead service line removed from a drinking water distribution system

A corroded lead service line was removed from a drinking water distribution system, and the microbial community was profiled using 16S rRNA gene techniques. This is the first report of the characterization of a biofilm on the surface of a corroded lead drinking water service line. The majority of phylotypes have been linked to heavy-metal-contaminated environments.     

Optimization of sulphide production in an anaerobic continuous biofilm process with sulphate reducing bacteria

Different biofilm reactors for sulphide production by sulphate reducing bacteria were compared in a packed bed reactor and in two suspended carrier biofilm reactors. Lactate was used as carbon source in the experiments. The process was reversibly inhibited by free sulphide at 14 mM. The packed bed reactor was more efficient and less sensitive to changes. The maximum load in the system was 5.3 g sulphate/l.d.     

Microbial community analysis of a full-scale DEMON bioreactor

Full-scale applications of autotrophic nitrogen removal technologies for the treatment of digested sludge liquor have proliferated during the last decade. Among these technologies, the aerobic/anoxic deammonification process (DEMON) is one of the major applied processes. This technology achieves nitrogen removal from wastewater through anammox metabolism inside a single bioreactor due to alternating cycles of aeration. To date, microbial community composition of full-scale DEMON bioreactors have never been reported. In this study, bacterial community structure of a full-scale DEMON bioreactor located at the Apeldoorn wastewater treatment plant was analyzed using pyrosequencing. This technique provided a higher-resolution study of the bacterial assemblage of the system compared to other techniques used in lab-scale DEMON bioreactors. Results showed that the DEMON bioreactor was a complex ecosystem where ammonium oxidizing bacteria, anammox bacteria and many other bacterial phylotypes coexist. The potential ecological role of all phylotypes found was discussed. Thus, metagenomic analysis through pyrosequencing offered new perspectives over the functioning of the DEMON bioreactor by exhaustive identification of microorganisms, which play a key role in the performance of bioreactors. In this way, pyrosequencing has been proven as a helpful tool for the in-depth investigation of the functioning of bioreactors at microbiological scale.     

The response of a bacterial biofilm community in a simulated industrial cooling water system to treatment with an anionic dispersant

The effect of a dispersant on the microbial community in a simulated open recirculating cooling water system was determined by continuous operation of the system over two consecutive periods of 196 and 252 d, respectively. An open recirculating cooling water system feeding a modified Robbin's Device with synthetic cooling water to simulate the environment of an industrial cooling water system was set up. Planktonic and biofilm (mild steel and Nylon(R)) samples were taken weekly in 1997 (196-d period) and fortnightly in 1998 (252-d period). Each biofilm was scraped off and diluted in 10-ml 1 x phosphate-buffered saline (PBS). Serial dilutions were performed and plated onto R2A agar (pH 8.0) to obtain the predominant culturable bacteria. The diversity was determined by allocating groups according to colony morphology, diameter and colour. Diversity was calculated according to the Shannon-Weaver Index. During the first run (1997), dispersant was added on day 57 to a final concentration of 15 mg l-1 for 49 d, stopped for 49 d and dosed at 30 mg l-1 for 41 d. The second run entailed adding dispersant to a final concentration of 30 mg l-1 on day 98 for 70 d, stopping dosing for 56 d and resuming dosing at 30 mg l-1 for another 28 d. The 2-year evaluation period demonstrated that the biofilm-removing action of the dispersant decreased to a point where it was not effective at all. Our results showed that the synthetic dispersant evaluated was only effective initially, but was ineffective in controlling biofouling on Nylon, and to a lesser degree on mild steel at the recommended (15 mg l-1) as well as at double the recommended concentration in the long term. The release of cells from biofilms observed when dispersant dosing was terminated, supports the notion that a community attaching in the presence of the surface active agent was selected for. The decreased efficacy may therefore be due to a selection of strains able to remain attached and/or attach in the presence of the dispersant as demonstrated by shifts in the biofilm communities on both Nylon and mild steel.     

Growth of legionella and other heterotrophic bacteria in a circulating cooling water system exposed to ultraviolet irradiation

The effect of ultraviolet irradiation on the growth and occurrence of legionella and other heterotrophic bacteria in a circulating cooling water system was studied. Water of the reservoir was circulated once in 28 h through a side-stream open channel u.v. radiator consisting of two lamps. Viable counts of legionellas and heterotrophic bacteria in water immediately after the u.v. treatment were 0—12 and 0·7—1·2% of those in the reservoir, respectively. U.v. irradiation increased the concentration of easily assimilable organic carbon. In the u.v. irradiated water samples incubated in the laboratory the viable counts of heterotrophic bacteria reached the counts in reservoir water within 5 d. The increase in viable counts was mainly due to reactivation of bacterialcells damaged by u.v. light, not because of bacterial multiplication. Despite u.v. irradiation the bacterial numbers in the reservoir water, including legionellas, did not decrease during the experimental period of 33 d. The main growth of bacteria in the reservoir occurred in biofilm and sediment, which were never exposed to u.v. irradiation.     

Microbial community and biomass characteristics associated severe membrane fouling during start-up of a hybrid anoxic-oxic membrane bioreactor

In MBR, severe membrane fouling is often observed in the initial phase in which biomass is yet fully acclimated and stabilized in terms of microbial community structure and biomass characteristics. The focus of this study was to investigate the microbial community development and its influence on biomass characteristics and membrane fouling during start-up of a hybrid anoxic-oxic MBR. PCR-DGGE analysis indicated that the microbial community shifted in start-up period when a severe membrane fouling was observed. Small particle size, high fractal dimension (DF) and high EPS production, which were closely associated with microbial community, were found to be the major contributors to the severe fouling. Microbial community development was most likely the ultimate factor responsible for the severe membrane fouling.     

全程硝化菌微生物学特性及在水处理领域的应用潜力

全程硝化菌是近期微生物氮循环领域的重大发现之一,引发了对其全球分布、系统发育特征和生理生化特性的广泛关注。本文综述了全程硝化菌在土壤、地表水、废水处理系统等生境的分布规律及影响因子;并从底物亲和力、代谢多样性等方面阐述了其与传统硝化微生物间的竞争互作和生态位分离机制;基于上述特征提出全程硝化菌在水处理领域中的应用前景,可能与其他脱氮微生物如反硝化菌、厌氧氨氧化菌和厌氧甲烷氧化菌等耦合实现在低氨氮、低溶解氧条件下的污水深度脱氮,从而节省能耗并降低温室气体排放。未来研究应继续深入研究全程硝化菌的生理生化特性,评价其生态功能和对氮素地球化学循环的贡献,并探索其在生物水处理等领域的应用潜力。