好氧反硝化细菌及其在微污染水源水修复中的应用研究进展

相比于氨氮,天然水体中的硝酸盐氮通常更稳定,导致更难将其从水中去除。由于好氧反硝化可以在有氧环境下进行反硝化作用去除硝酸盐氮,该过程对含有较高溶解氧的天然水体中硝酸盐氮处理有重要作用。本文综述了好氧反硝化菌的分离纯化现状、微生物代谢机制和环境影响因子,并介绍了功能菌群在微污染饮用水源水生物修复的应用研究进展。与一般的厌氧反硝化类似,好氧反硝化菌的种属分布较广,常见的如假单胞菌属(Pseudomoas)、产碱杆菌属(Alcaligenes)、副球菌属(Paracoccus)和芽孢杆菌属(Bacillus)等所属部分微生物均有好氧反硝化能力。大部分好氧反硝化菌株在最佳生长条件下(25–37℃、溶解氧浓度为3–5mg/L、pH为7–8、碳氮比为5–10)具有高效的脱氮效率。但目前好氧反硝化作用在微污染饮用水源水的生物修复方面的应用仍有着脱氮性能不稳定、菌剂流失等不足。此外,目前较少相关中试及实际工程应用的研究,需要进一步的深入探究。     

Metabolic diversity of aromatic hydrocarbon-degrading bacteria from a petroleum-contaminated aquifer

We characterized bacteria from contaminated aquifers for their ability to utilize aromatic hydrocarbons under hypoxic (oxygen-limiting) conditions (initial dissolved oxygen concentration about 2 mg/l) with nitrate as an alternate electron acceptor. This is relevant to current intense efforts to establish favorable conditions forin situ bioremediation. Using samples of granular activated carbon slurries from an operating groundwater treatment system, we isolated bacteria that are able to use benzene, toluene, ethylbenzene, orp-xylene as their sole source of carbon under aerobic or hypoxic-denitrifying conditions. Direct isolation on solid medium incubated aerobically or hypoxically with the substrate supplied as vapor yielded 10³ to 10⁵ bacteria ml^–1 of slurry supernatant, with numbers varying little with respect to isolation substrate or conditions. More than sixty bacterial isolates that varied in colony morphology were purified and characterized according to substrate utilization profiles and growth condition (i.e., aerobic vs. hypoxic) specificity. Strains with distinct characteristics were obtained using benzene compared with those isolated on toluene or ethylbenzene. In general, isolates obtained from direct selection on benzene minimal medium grew well under aerobic conditions but poorly under hypoxic conditions, whereas many ethylbenzene isolates grew well under both incubation conditions. We conclude that the conditions of isolation, rather than the substrate used, will influence the apparent characteristic substrate utilization range of the isolates obtained. Also, using an enrichment culture technique, we isolated a strain ofPseudomonas fluorescens, designated CFS215, which exhibited nitrate dependent degradation of aromatic hydrocarbons under hypoxic conditions.Abbreviations BTEX benzene, toluene, ethylbenzene, andp-xylene - HPLC high performance liquid chromatography - GAC granular activated carbon     

Effect of oxygen on denitrification in continuous chemostat culture withComamonas sp SGLY2

Continuous cultures ofComamonas sp SGLY2 were grown anaerobically prior to establishing steady states at different oxygen flow rates. At a low oxygen transfer rate, no dissolved oxygen accumulated in the medium and all nitrate was reduced to dinitrogen. Concurrently with the increase of dissolved oxygen concentration in the liquid phase, the rate of denitrification decreased. However, at a dissolved oxygen concentration near saturation (33 mg L^–1), a part of the electron flow always diverted to nitrate with production of dinitrogen: the aerobic denitrification rate was equivalent to 35% of that calculated under anaerobic conditions. These experiments reflected the co-utilization of oxygen and N-oxides and the production of dinitrogen, up to saturated conditions, which implied synthesis and activity of the four denitrifying enzymes under various aeration conditions.     

Soil nitrogen transformations and nitrate utilization by Deschampsia flexuosa (L.) Trin. at two contrasting heathland sites

Two Dutch heathland sites Hoorneboeg (HB) and Ede, dominated by Deschampsia flexuosa and differing in nitrate production, were sampled for an entire growing season. A large number of soil and plant parameters were monitored in an attempt to assess the contribution of nitrate in the N supply and its assimilation by Deschampsia.Average NO₃ ^– and NH₄ ⁺ concentrations (mg kg^–1) in the top 10-cm depth were 0.03 and 2.2, respectively, for HB, and 2.1 and 6.7, respectively, for Ede. Laboratory incubations of intact cores and experiments with FH-layer suspensions showed significantly higher mineralization and nitrification rates for the Ede site during most of the season. Nitrification was largely controlled by the rate of net N-mineralization, which in turn was highly affected by soil moisture. Nitrate production was virtually zero at HB and accounted for 25% of the net N-mineralization at Ede.Shoot chemical composition showed no essential differences for the two sites, but mean in vivo (current) foliar NRA was almost 2-fold higher at Ede than at HB, indicating some utilization of nitrate at the former location. At the HB site with essentially no nitrate production, however, enzyme activities were clearly higher than basal constitutive levels in NH₄ ⁺-fed plants. Apparently, shoot NRA at the HB site became positively affected by factors other than nitrate availability and/or showed disproportional increases in response to atmospheric nitrate inputs. Root NRA displayed the same low basal level at the two sites. Nitrate fertilization (100 kg N ha^–1) yielded maximally induced foliar NRAs similar to levels found in hydroponic nitrate plants. Although no accumulation of free NO₃ ^– was observed in shoots from fertilized plots, increases in foliar concentrations of both organic N and carboxylates clearly indicated nitrate assimilation. Root NRA showed no response to nitrate addition.It is concluded that current NRA measurements in Deschampsia at heathland sites are of limited value only, especially when interpreted in isolation. A combined approach, using concurrently conducted soil and plant analyses, will allow the extent of nitrate utilization in the field to be best characterized.Publication 2013 of the Netherlands Institute of Ecology.FAX no corresponding author: +31 8306 23227     

Seasonal flooding,nitrogen mineralization and nitrogen utilization in a prairie marsh

Flooding can be an important control of nitrogen (N) biogeochemistry in wetland ecosystems. In North American prairie marshes, spring flooding is a dominant feature of the physical environment that increases emergent plant production and could influence N cycling. I investigated how spring flooding affects N availability and plant N utilization in whitetop (Scolochloa festucacea) marshes in Manitoba, Canada by comparing experimentally spring-flooded marsh inside an impoundment with adjacent nonflooded marsh. The spring-flooded marsh had net N mineralization rates up to 4 times greater than nonflooded marsh. Total growing season net N mineralization was 124 kg N ha^–1 in the spring-flooded marsh compared with 62 kg N ha^–1 in the nonflooded marsh. Summer water level drawdown in the spring-flooded marsh decreased net N mineralization rates. Net nitrification rates increased in the nonflooded marsh following a lowering of the water table during mid summer. Growing season net nitrification was 33 kg N ha^–1 in the nonflooded marsh but < 1 kg N ha^–1 in the spring-flooded marsh. Added NO₃ ^–1 induced nitrate reductase (NRA) activity in whitetop grown in pot culture. Field-collected plants showed higher NRA in the nonflooded marsh. Nitrate comprised 40% of total plant N uptake in the nonflooded marsh but <1% of total N uptake in the spring-flooded marsh. Higher plant N demand caused by higher whitetop production in the spring-flooded marsh approximately balanced greater net N mineralization. A close association between the presence of spring flooding and net N mineralization and net nitrification rates indicated that modifications to prairie marshes that change the pattern of spring inundation will lead to rapid and significant changes in marsh N cycling patterns.     

Changes in heated and autoclaved forest soils of S.E. Australia. I. Carbon and nitrogen

The effect of heating and autoclaving on extractable nitrogen, N mineralisation and C metabolism was studied by heating five forest soils in the laboratory, simulating the range of effects of heat due to bushfire. Top soil (0–5 cm) was heated to 60 °C, 120 °C and 250 °C for 30 minutes; unheated soil was taken as a control. Samples of the soil heated to 250 °C were also inoculated with fresh soil to accelerate the recovery of the microbial population. Soil autoclaving was carried out as another heat treatment (moist heat). Soils were analysed immediately after heating and 3 times during seven months of incubation to assess immediate and longer-term effects of heating.Extractable N (organic and mineral forms) increased after heating to 120 °C, but decreased with further heating to 250 °C suggesting the volatilisation of N. N associated with microbial biomass diminished with heating and was barely detectable after the 250 °C treatment. Microbial biomass was an important source of soluble N in heated soils, and only partly recovered during subsequent long incubation. The amount of N mineralised during incubation depended on both soil and temperature. Nitrification did not occur when soils were heated to 250 °C (with or without inoculum), or after autoclaving, demonstrating the high sensitivity of nitrifiers to heat. At the beginning of soil incubation, respiration was enhanced in heated soils (250 °C, 250 °C inoculated) and autoclaved soils, but after 30 days of incubation respiration decreased to values either similar to or lower than those in control. This respiration pattern indicated that a fraction of labile C was released by heating, which was quickly mineralised within 30 days of incubation. These results demonstrate some effects of soil heating on C and N dynamics in forest soils.     

Effects of diffusion limitation on immobilized nitrifying microorganisms at low temperatures

Activation energies of suspended and immobilized nitrifying bacteria were determined and compared to determine if diffusion limitation results in decreased sensitivity for temperature. The activation energy for the respiration activity of suspended Nitrosomonas europaea and Nitrobacter agilis was found to be 86.4 and 58.4 kJ mol(-1), respectively. The activation energy for oxygen diffusion in the support material, kappa-carrageenan, determined from the effect of temperature on the effective diffusion coefficient (D), was 17.2 kJ mol(-1). Consequently, the apparent actvation energy of diffusion limited cells should be lower. It was indeed shown that due to the effect of diffusion limitation and to temperature effects on the Monod constant K(s), the immobilized-cell activity was less sensitive to temperature. The apparent activation energy for immobilized Ns. europaea was between 28.6 and 94.2 kJ mol(-1) and for immobilized Nb. agilis between 1.4 and 72.9 kJ mol(-1), depending on the oxygen concentration and temperature. (c) 1995 John Wiley & Sons, Inc.     

Flatworms flatten to size up each other

When copulations are costly, fertilization is reciprocal and fecundity is positively related to size, hermaphrodites are expected to favour large partners, leading to size-assortative mating. Size-related mate choice has, however, never been observed in hermaphrodites. In the flatworm Dugesia gonocephala copulations cost time and are reciprocal, and size is a positive predictor of female fecundity. Every copulation is preceded by a phase in which one partner glides on top of the other and both spread out and flatten, suggesting that partners assess each other''s size. A total of 124 copulating pairs collected on four different dates, proved that mating is size-assorted in the field. In experiments with groups, more copulations took place between equally sized individuals than were expected when matings were random. In experiments with pairs, partners of different size exhibited twice as many mating attempts for the first copulation than did partners of the same size. We conclude that D. gonocephala employs a unique kind of pre-copulatory ''flattening'' behaviour as a mechanism to signal as well as to assess relative size. This does not only confirm that hermaphrodites can mate assortatively when certain assumptions are met, it also proves that even lower invertebrates can show active mate choice.