Dynamics of specific ammonia-oxidizing bacterial populations and nitrification in response to controlled shifts of ammonium concentrations in wastewater.

Ammonia-oxidizing bacteria (AOB) are essential for the nitrification process in wastewater treatment. To retain these slow-growing bacteria in wastewater treatment plants (WWTPs), they are often grown as biofilms, e.g., on nitrifying trickling filters (NTFs) or on carriers in moving bed biofilm reactors (MBBRs). On NTFs, a decreasing ammonium gradient is formed because of the AOB activity, resulting in low ammonium concentrations at the bottom and reduced biomass with depth. To optimize the NTF process, different ammonium feed strategies may be designed. This, however, requires knowledge about AOB population dynamics. Using fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy, we followed biomass changes during 6 months, of three AOB populations on biofilm carriers. These were immersed in aerated MBBR tanks in a pilot plant receiving full-scale wastewater. Tanks were arranged in series, forming a wastewater ammonium gradient mimicking an NTF ammonium gradient. The biomass of one of the dominating Nitrosomonas oligotropha-like populations increased after an ammonium upshift, reaching levels comparable to the high ammonium control in 28 days, whereas a Nitrosomonas europaea-like population increased relatively slowly. The MBBR results, together with competition studies in NTF systems fed with wastewater under controlled ammonium regimes, suggest a differentiation between the two N. oligotropha populations, which may be important for WWTP nitrification.

Nitrification potential and population dynamics of nitrifying bacterial biofilms in response to controlled shifts of ammonium concentrations in wastewater trickling filters.

Nitrogen removal in wastewater treatment is energy consuming and often carried out in biofilm nitrifying trickling filters (NTFs). We investigated nitrification potential and population dynamics of nitrifying bacteria in pilot-plant NTFs fed with full-scale plant wastewater with high (8-9 mg NH(4)(+)l(-1)) or low (<0.5mg NH(4)(+)l(-1)) ammonium concentrations. After ammonium shifts, nitrification potentials stabilized after 10-43 days depending on feed regime. An NTF fed with 3 days of high, and 4 days of low load per week reached a high nitrification potential, whereas a high load for 1 day a week gave a low potential. Nitrosomonas oligotropha dominated the AOB and changes in nitrification potentials were not explained by large population shifts to other AOBs. Although nitrification potentials were generally correlated with the relative amounts of AOB and NOB, this was not always the case. Ammonium feed strategies can be used to optimize wastewater treatment performance.

Community survey of ammonia-oxidizing bacteria in full-scale activated sludge processes with different solids retention time.

AIMS: To study the effects of different solids retention time (SRT) on the nitrification activity and community composition of ammonia-oxidizing bacteria (AOB) in two full-scale activated sludge processes during a 5-month period. METHODS AND RESULTS: The AOB community composition was analysed using fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE), and the identified populations were enumerated by quantitative FISH. Potential nitrification rates were determined in batch tests and the in situ rates were calculated from mass balances of nitrogen in the plants. Increased SRT reduced the nitrification activity, but neither the number per mixed liquor suspended solids nor community composition of AOB were affected. Two dominant AOB populations related to Nitrosomonas europaea and Nitrosomonas oligotropha were identified by FISH, whereas only the latter could be detected by DGGE. CONCLUSIONS: The effect of a longer SRT on the activity was probably because of physiological changes in the AOB community rather than a change in community composition. SIGNIFICANCE AND IMPACT OF THE STUDY: Physiological alterations of a stable AOB community are possible and may stabilize activated sludge processes. The commonly used FISH probes designed to target all beta-proteobacterial AOB does not detect certain Nitrosomonas oligotropha populations, leading to an underestimation of AOB if a wider set of probes is not used.

Inhibition of bacterial alpha-glucosidases by castanospermine in pure cultures and activated sludge.

Castanospermine (CAST) is a known and potent inhibitor of various alpha-glucosidases in eukaryotes. In this work, we elucidated whether CAST could also be used for determining bacterial alpha-glucosidase activity, when measured with 4-methylumbelliferyl-alpha- D-glucoside as a substrate, both in a complex bacterial community, in activated sludge and in pure cultures of bacterial isolates. We found that 140 microM CAST inhibited alpha-glucosidase activity by 30% in a pure culture of Pseudomonas stutzeri. The alpha-glucosidase activity in Chryseobacterium gleum was inhibited by 90% at a concentration of 150 microM CAST, whereas the alpha-glucosidase in Paracoccus denitrificans was resistant to the inhibitor. CAST (140 microM) reduced alpha-glucosidase activity in activated sludge by 40%, the respiration rate being reduced by only 12%. No significant inhibition of the respiration rate was observed in Ps. stutzeri or Pa. denitrificans, whereas the respiration rate in C. gleum grown in a medium containing starch was inhibited by 50% with 140 microM CAST. No effect of CAST was observed in C. gleum grown in a complex medium. This indicated that CAST, at the concentration used, did not cause a general negative effect on bacterial activity. The results suggest that the CAST assay may potentially be useful in determining whether alpha-glucosidase activity, starch, poly- and disaccharides contribute appreciably to the overall activity of a bacterial community. However, the assay should not be used for quantitative measurements of such activity.

Assay for determination of alpha-glucosidase and peptidase activity and location in a nitrifying trickling filter.

Enzymatic alpha-glucosidase and peptidase activity in a nitrifying trickling filter (NTF) at the Rya wastewater treatment plant, Göteborg, Sweden, was investigated to evaluate whether these activities can be used as indicators of heterotrophic activity and polymer degradation. Samples of the biofilm were taken from the NTF and incubated in sterile filtered effluent water from the NTF with the addition of soluble starch, peptone, and ammonium chloride. In order to determine the distribution of enzyme activities, the alpha-glucosidase and peptidase activities were measured in the biofilm samples, in the filtered effluent water from the NTF and in the water phase in which the biofilm was incubated. Activities of both enzymes were found both in the effluent water from the NTF and in the biofilm. The enzyme activities were elevated in the samples when starch and peptone were present. In addition, there was a significant inhibition of ammonium oxidation in samples incubated with starch and peptone. Thus, the presence of starch, peptone and ammonium resulted in increased activity of heterotrophs, which lead to an inhibition of the nitrifiers, probably via competition for available oxygen.

New procedure for extraction of ammonium from natural waters for N isotopic ratio determinations.

A new method for extracting ammonium from natural waters for N isotopic ratio determination is described. The method employs the conversion of the ammonium nitrogen into indophenol, which is then concentrated onto an octadecylsilane column. The method shows accuracy and precision comparable to those of other methods described in the literature. Some results from field experiments on the Swedish west coast are presented.

Denitrification rates in the low-oxygen waters of the stratified baltic proper.

Denitrification activity was shown in the deep, low-oxygen waters of the Baltic proper by both in vitro and in situ methods. The vertical distribution of NO(3) in the water column showed nitrate consumption and NO(2) and N(2)O maxima in the deep waters when O(2) was below 0.2 ml liter, which is suggestive evidence for denitrification. Direct in situ evidence for denitrification was obtained by finding an N(2) saturation of up to 108% in the deep waters. When these waters were incubated with NO(3), N(2) was produced. Quantification of the denitrification rate done by the addition of C(2)H(2) to water samples from the active depths showed a rate of about 0.10 mumol liter day.