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Nitrification at Low pH by Aggregated Chemolithotrophic Bacteria.

A study was performed to gain insight into the mechanism of acid-tolerant, chemolithotrophic nitrification. Microorganisms that nitrified at pH 4 were enriched from two Dutch acid soils. Nitrate production in the enrichment cultures was indicated to be of a chemolithoautotrophic nature as it was (i) completely inhibited by acetylene at a concentration as low as 1 mumol/liter and (ii) strongly retarded under conditions of carbon dioxide limitation. Electron microscopy of the enrichment cultures showed the presence of bacteria that were morphologically similar to strains of known chemolithotrophic nitrifying genera. Many of the enriched bacteria, in particular those that were identified as ammonium oxidizers, were aggregated. Filtration experiments indicated that aggregated cells were able to nitrify at low pH, whereas single cells were not. It is hypothesized that cells inside the aggregates are protected against the toxicity of nitrous acid. Nitrification by aggregated chemolithoautotrophic bacteria may be the dominating process of nitrate formation in many acid soils as it does not appear to depend on the existence of microsites of high pH (acid-sensitive autotrophic nitrification) or on the availability of organic carbon (heterotrophic nitrification).

Journal Article↗

Pathways of inorganic nitrogen assimilation in chemoautotrophic bacteria-marine invertebrate symbioses: expression of host and symbiont glutamine synthetase

Symbioses between chemoautotrophic bacteria and marine invertebrates living at deep-sea hydrothermal vents and other sulfide-rich environments function autotrophically by oxidizing hydrogen sulfide as an energy source and fixing carbon dioxide into organic compounds. For chemoautotrophy to support growth, these symbioses must be capable of inorganic nitrogen assimilation, a process that is not well understood in these or other aquatic symbioses. Pathways of inorganic nitrogen assimilation were investigated in several of these symbioses: the vent tubeworms Riftia pachyptila and Tevnia jerichonana, the vent bivalves Calyptogena magnifica and Bathymodiolus thermophilus, and the coastal bivalve Solemya velum. Nitrate reductase activity was detected in R. pachyptila, T. jerichonana and B. thermophilus, but not in C. magnifica and S. velum. This is evidence for nitrate utilization, either assimilation or respiration, by some vent species and is consistent with the high levels of nitrate availability at vents. The ammonia assimilation enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were detected in all symbioses tested, indicating that ammonia resulting from nitrate reduction or from environmental uptake can be incorporated into amino acids. A complicating factor is that GS and GDH are potentially of both host and symbiont origin, making it unclear which partner is involved in assimilation. GS, which is considered to be the primary ammonia-assimilating enzyme of autotrophs, was investigated further. Using a combination of molecular and biochemical approaches, host and symbiont GS were distinguished in the intact association. On the basis of Southern hybridizations, immunoreactivity, subunit size and thermal stability, symbiont GS was found to be a prokaryote GS. Host GS was distinct from prokaryote GS. The activities of host and symbiont GS were separated by anion-exchange chromatography and quantified. Virtually all activity in symbiont-containing tissue was due to symbiont GS in R. pachyptila, C. magnifica and B. thermophilus. In contrast, no symbiont GS activity was detected in the gill of S. velum, the predominant activity in this species appearing to be host GS. These findings suggest that ammonia is primarily assimilated by the symbionts in vent symbioses, whereas in S. velum ammonia is first assimilated by the host. The relationship between varying patterns of GS expression and host-symbiont nutritional exchange is discussed.

Journal Article↗

The expanded bed biofilter: combined nitrification, solids destruction, and removal of bacteria.

Developed for tertiary nitrification, this biofilter also removed carbonaceous BOD (cBOD) and (SS). Because the biofilter is expanded, it cannot clog, and therefore does not require backflushing; yet, it removed a significant proportion of the influent SS. This unanticipated capability was due to the activities of heterotrophic bacteria, protozoa, and metazoa (nematode and oligochaete worms). The expanded bed is an intensified process, which is based on natural immobilization of microbes to small support particles. Using glassy coke as the support material, an attached layer of microbes develops, forming particulate biofilms having a superficial surface area of 1 800 m2 m(-3)(expandedbed). Autotrophic nitritifiers (Nitrosomonas spp.) were detected in the biofilm using rRNA-based molecular methods and were likely responsible, at least in part, for reducing the ammonia concentration by up to 99% (to 0.1 mg L(-1)), while the other organisms reduced cBOD and SS by up to 56% and 62%, respectively. Furthermore, the influent concentrations of Escherichia coli, coliform and heterotrophic bacteria were reduced by over 80%. It thereby provides a single process solution for combined tertiary nitrification and solids removal. Operating the process to consistently achieve < 0.5mg NH3N L(-1) and at the same time removing a significant fraction of cBOD and SS, it can replace processes such as SAFs or NTFs followed by a sandfilter.

Ammonia↗

Heavy metal toxicity to microbe-mediated ecologic processes: a review and potential application to regulatory policies.

Microorganisms are sensitive to heavy metal pollution as are other components of the biota. However, most studies on the interactions between microbes and heavy metals have been conducted in synthetic media or in altered (e.g., sterilized) environmental samples and usually have used only single species. Few studies have evaluated the effects of heavy metals on the activities of natural heterogeneous microbial populations, both autotrophic and heterotrophic, in terrestrial and aquatic environments. These latter studies have shown that heavy metals inhibit primary productivity, nitrogen fixation, the mineralization of carbon, nitrogen, sulfur, and phosphorus, litter decomposition, and enzyme synthesis and activity in soils, sediments, and surface waters. The potential adverse effects of heavy metals on such microbe-mediated ecologic processes need to be incorporated into the methodologies used by regulatory agencies, such as the U.S. Environmental Protection Agency, to prepare environmental risk assessments which, in turn, are used to formulate environmental criteria, such as the Water Quality Criteria, and to evaluate the safety to the environment of exposure to "new chemical substances," as mandated by the U.S. Toxic Substances Control Act of 1976. To provide appropriate data that can be assimilated into regulatory policy, it is essential that microbial ecotoxicity tests be standardized, are neither costly nor difficult to train personnel to conduct, and produce data that can be quantitated.

Bacteria↗

Tree-girdling to separate root and heterotrophic respiration in two Eucalyptus stands in Brazil.

The release of carbon as CO2 from belowground processes accounts for about 70% of total ecosystem respiration. Insights about factors controlling soil CO2 efflux are constrained by the challenge of apportioning sources of CO2 between autotrophic tree roots (and mycorrhizal fungi) and heterotrophic microorganisms. In some temperate conifer forests, the reduction in soil CO2 efflux after girdling (phloem removal) has been used to separate these sources. Girdling stops the flow of carbohydrates to the belowground portion of the ecosystem, which should slow respiration by roots and mycorrhizae while heterotrophic respiration should remain constant or be enhanced by the decomposition of newly dead roots. Therefore, the reduction in CO2 efflux after girdling should be a conservative estimate of the belowground flux of C from trees. We tested this approach in two tropical Eucalyptus plantations. Tree canopies remained intact for more than 3 months after girdling, showing no reduction in light interception. The reduction in soil CO2 efflux averaged 16-24% for the 3-month period after girdling. The reduction in CO2 efflux was similar for plots with one half of the trees girdled and those with all of the trees girdled. Girdling did not reduce live fine root biomass for at least 5 months after treatment, indicating that large reserves of carbohydrates in the root systems of Eucalyptus trees maintained the roots and root respiration. Our results suggest that the girdling approach is unlikely to provide useful insights into the contribution of tree roots and heterotrophs to soil CO2 efflux in this type of forest ecosystem.

Brazil↗

Whole-lake carbon-13 additions reveal terrestrial support of aquatic food webs.

Ecosystems are supported by organic carbon from two distinct sources. Endogenous carbon is produced by photosynthesis within an ecosystem by autotrophic organisms. Exogenous carbon is produced elsewhere and transported into ecosystems. Consumers may use exogenous carbon with consequent influences on population dynamics, predator-prey relationships and ecosystem processes. For example, exogenous inputs provide resources that may enhance consumer abundance beyond levels supported by within-system primary production. Exogenous fluxes of organic carbon to ecosystems are often large, but this material is recalcitrant and difficult to assimilate, in contrast to endogenously produced organic matter, which is used more easily. Here we show, by the experimental manipulation of dissolved inorganic (13)C in two lakes, that internal primary production is insufficient to support the food webs of these ecosystems. Additions of NaH(13)CO(3) enriched the (13)C content of dissolved inorganic carbon, particulate organic carbon, zooplankton and fish. Dynamics of (13)C indicate that 40-55% of particulate organic carbon and 22-50% of zooplankton carbon are derived from terrestrial sources, showing that there is significant subsidy of these ecosystems by organic carbon produced outside their boundaries.

Animals↗

[Role of exogenous carbon dioxide in the metabolism of methane-oxidizing bacteria].

The object of this work was to study the ability of methane oxidizing bacteria to use CO2 as an acceptor of electrons liberated in methane oxidation and the role of CO2 fixation in the constructive metabolism of the bacteria. All of the studied methane oxidizing bacterial cultures were found to be capable of fixing the 14C of hydrocarbonate. The activity of the process was shown to be similar in different strains. Up to 30% of the carbon in the biomass composition could originate from the carbon of HCO3-. Methane oxidizing bacteria that assimilated C1-compounds via the hexulose phosphate and serine pathways had the same level of HCO3- fixation. No differences were found among strains of the same species, among species, or among genera. The assimilation of HCO3- was catalyzed by PEP-carboxylase (i. e. in a heterotrophous way) or, in some cultures, by ribulose-1,5-diphosphate carboxylase, the key enzyme in the autotrophous pathway of CO2 assimilation. The enzymological mechanisms of HCO3- assimilation are discussed. The biological role of CO2 fixation in the metabolism of methane oxidizing bacteria that use the hexulose phosphate and serine pathways of methane assimilation may be different. The process can either play the role of anapleurotic reactions in the tricarboxylic acid cycle, or be an element of the serine pathway of methane assimilation. Calculations have shown that the extent to which a substrate to be metabolized is reduced seems to determine the activity of exogenous CO2 fixation. The contribution made by HCO3- fixation into the carbon metabolism of methane oxidizing bacteria confirms that they are related to lithotrophous organisms.

Bicarbonates↗

Kinetic model of a granular sludge SBR: influences on nutrient removal.

A mathematical model was developed that can be used to describe an aerobic granular sludge reactor, fed with a defined influent, capable of simultaneously removing COD, nitrogen and phosphate in one sequencing batch reactor (SBR). The model described the experimental data from this complex system sufficiently. The effect of process parameters on the nutrient removal rates could therefore be reliably evaluated. The influence of oxygen concentration, temperature, granule diameter, sludge loading rate, and cycle configuration were analyzed. Oxygen penetration depth in combination with the position of the autotrophic biomass played a crucial role in the conversion rates of the different components and thus on overall nutrient removal efficiencies. The ratio between aerobic and anoxic volume in the granule strongly determines the N-removal efficiency as it was shown by model simulations with varying oxygen concentration, temperature, and granule size. The optimum granule diameter for maximum N- and P-removal in the standard case operating conditions (DO 2 mg L(-1), 20 degrees C) was found between 1.2 and 1.4 mm and the optimum COD loading rate was 1.9 kg COD m(-3) day(-1). When all ammonia is oxidized, oxygen diffuses to the core of the granule inhibiting the denitrification process. In order to optimize the process, anoxic phases can be implemented in the SBR-cycle configuration, leading to a more efficient overall N-removal. Phosphate removal efficiency mainly depends on the sludge age; if the SRT exceeds 30 days not enough biomass is removed from the system to keep effluent phosphate concentrations low.

Ammonia↗

Factors influencing stable isotope ratios in CH4 and CO2 within subenvironments of freshwater wetlands: implications for delta-signatures of emissions.

Much uncertainty still exists regarding spatial and temporal variability of stable isotope ratios (13C/12C and D/H) in different CH4-emission sources. Such variability is especially prevalent in freshwater wetlands where a range of processes can influence stable isotope compositions, resulting in variations of up to approximately 50% for delta13C-CH4 and approximately 50% for deltaD-CH4 values. Within a temperate-zone bog and marsh situated in southwestern Ontario, Canada, gas bubbles in pond sediments exhibit only minor seasonal and spatial variation in delta13C-CH4, deltaD-CH4 and delta13C-CO2 values. In pond sediments, CO2 appears to be the main source of carbon during methanogenesis either directly via CO2 reduction or indirectly through dissimilation of autotrophic acetate. In contrast, CH4 production occurs primarily via acetate fermentation at shallow depths in peat soils adjacent to ponds at each wetland. At greater depths within soils, sigmaCO2 and H2O increasingly exert an influence on delta13C- and deltaD-CH4 values. Secondary alteration processes (e.g., methanotrophy or diffusive transport) are unlikely to be responsible for depth-related changes in stable isotope values of CH4. Recent models that attempt to predict deltaD-CH4 values in freshwater environments from D/H ratios in local precipitation do not adequately account for such changes with depth. Subenvironments (i.e., soil-forming and open water areas) in wetlands should be considered separately with respect to stable isotope signatures in CH4 emission models.

Atmosphere↗

Microbial succession and assembly shaped by sulfur, spatial partitioning, and water flow in a volcanic acidic river of northern Patagonia.

Extreme acidic environments represent natural laboratories for investigating the mechanisms of microbial community assembly, yet the ecological processes structuring these communities remain incompletely understood. Here, we investigate how spatial partitioning, hydrodynamics, and colonization history shape microbial succession in a unique sulfur-rich, acidic river of volcanic origin in northern Patagonia. We combined 16S rRNA gene profiling and shotgun metagenomics with a multi-scale experimental framework encompassing water column fractionation and colonization assays under native and controlled conditions. Microbial diversity was strongly influenced by spatial fractionation, with free-living communities exhibiting higher richness and temporal variability than particle-associated assemblages. Water flow modulated community structure, increasing evenness in free-living fractions under high-flow conditions, but had limited impact on particle-attached communities. Colonization of sulfur-beads followed a structured successional trajectory, with autotrophic sulfur oxidizers dominating early stages and heterotrophs adapted to biofilm lifestyles increasing over time. Ex situ recolonization assays revealed strong priority effects, with initial colonizers determining successional trajectories. Turnover analyses revealed that the balance among stochastic and deterministic assembly processes shifted across communities with pronounced stochasticity in the water column and flow-dependent effects in free-living communities, while biofilm associated communities on sulfur-beads exhibited stronger contribution of deterministic selection. These ecological patterns were mirrored by functional differentiation, with gene enrichment analyses revealing adaptive signatures of substrate attachment and resource acquisition. By integrating fine-scale environmental variation with colonization dynamics, this study reveals how microscale habitat structure and temporal fluxes jointly modulate microbial community assembly rules, offering a nuanced framework to dissect ecological processes in extreme systems.

Sulfur↗

Perchlorate reduction by autotrophic bacteria in the presence of zero-valent iron.

A series of batch experiments were performed to study the combination of zero-valent iron (ZVI) with perchlorate-reducing microorganisms (PRMs) to remove perchlorate from groundwater. In this method, H2 produced during the process of iron corrosion by water is used by PRMs as an electron donor to reduce perchlorate to chloride. Perchlorate degradation rates followed Monod kinetics, with a normalized maximum utilization rate (rmax) of 9200 microg g(-1) (dry wt) h(-1) and a half-velocity constant (Ks) of 8900 microg L(-1). The overall rate of perchlorate reduction was affected by the biomass density within the system. An increase in the OD600 from 0.025 to 0.08 led to a corresponding 4-fold increase of perchlorate reduction rate. PRM adaptation to the local environment and initiation of perchlorate reduction was rapid under neutral pH conditions. At the initial OD600 of 0.015, perchlorate reduction followed pseudo-first-order reaction rates with constants of 0.059 and 0.033 h(-1) at initial pH 7 and 8, respectively. Once perchlorate reduction was established, the bioreductive process was insensitive to the increases of pH from near neutral to 9.0. In the presence of nitrate, perchlorate reduction rate was reduced, but not inhibited completely.

Hydrogen↗

Flow cytometric analysis of fine particles in a eutrophic lake.

Fine particles play an important role, not only in aquatic biogeochemical processing but also in the distribution, transfer and transformation of pollutants in the aquatic environment. Flow cytometry, widely used in biomedical research, allows fast counting and optical analysis of individual particles. Organic autotrophic particles contain naturally fluorescing pigments, such as chlorophyll and phycoerythrin. Different populations have different sizes and pigments. They also have different ratios of pigments. In general, side angle scatter (SSC) is related to the size, shape and refractive index of particles. When a 488 nm wavelength was used to excite chlorophyll and phycoerythrin fluorescence, the pigments of organic autotrophic particles emitted red and orange light. Fine particles were detected by flow cytometry (FCM) in the southern part of a eutrophic lake in winter. We found that organic autotrophic particles belonged to three populations, which represented only 15.89% of total fine particles. Organic non-living particles and inorganic particles represented the greater part (84.11%) of total fine particles. This study also demonstrated that flow cytometry is well suited to the dynamic monitoring and analysis of natural water aquatic particles that were difficult to study with traditional methods.

Chlorophyll↗

Adaptation and acclimation of photosynthetic microorganisms to permanently cold environments.

Persistently cold environments constitute one of our world's largest ecosystems, and microorganisms dominate the biomass and metabolic activity in these extreme environments. The stress of low temperatures on life is exacerbated in organisms that rely on photoautrophic production of organic carbon and energy sources. Phototrophic organisms must coordinate temperature-independent reactions of light absorption and photochemistry with temperature-dependent processes of electron transport and utilization of energy sources through growth and metabolism. Despite this conundrum, phototrophic microorganisms thrive in all cold ecosystems described and (together with chemoautrophs) provide the base of autotrophic production in low-temperature food webs. Psychrophilic (organisms with a requirement for low growth temperatures) and psychrotolerant (organisms tolerant of low growth temperatures) photoautotrophs rely on low-temperature acclimative and adaptive strategies that have been described for other low-temperature-adapted heterotrophic organisms, such as cold-active proteins and maintenance of membrane fluidity. In addition, photoautrophic organisms possess other strategies to balance the absorption of light and the transduction of light energy to stored chemical energy products (NADPH and ATP) with downstream consumption of photosynthetically derived energy products at low temperatures. Lastly, differential adaptive and acclimative mechanisms exist in phototrophic microorganisms residing in low-temperature environments that are exposed to constant low-light environments versus high-light- and high-UV-exposed phototrophic assemblages.

Acclimatization↗

Carboxyl-terminal processing of the cytoplasmic NAD-reducing hydrogenase of Alcaligenes eutrophus requires the hoxW gene product.

Two open reading frames (ORFs) were identified immediately downstream of the four structural genes for the soluble hydrogenase (SH) of Alcaligenes eutrophus H16. While a mutation in ORF2 had no obvious effect on hydrogen metabolism, an in-frame deletion in ORF1, subsequently designated hoxW, led to a complete loss of SH activity and hence a significant retardation of autotrophic growth on hydrogen. Hydrogen oxidation in the hoxW mutant was catalyzed by the second hydrogenase, a membrane-bound enzyme. Assembly of the four subunits of the SH was blocked in mutant cells, and HoxH, the hydrogen-activating subunit, accumulated as a precursor which was still capable of binding nickel. Protein sequencing revealed that HoxH isolated from the wild type terminates at His-464, whereas the C-terminal amino acid sequence of HoxH from the hoxW mutant is colinear with the deduced sequence. Processing of the HoxH precursor was restored in vitro by a cell extract containing HoxW. These results indicate that HoxW is a highly specific carboxyl-terminal protease which releases a 24-amino-acid peptide from HoxH prior to progression of subunit assembly.

Alcaligenes↗

Harmful algal blooms: causes, impacts and detection.

Blooms of autotrophic algae and some heterotrophic protists are increasingly frequent in coastal waters around the world and are collectively grouped as harmful algal blooms (HABs). Blooms of these organisms are attributed to two primary factors: natural processes such as circulation, upwelling relaxation, and river flow; and, anthropogenic loadings leading to eutrophication. Unfortunately, the latter is commonly assumed to be the primary cause of all blooms, which is not the case in many instances. Moreover, although it is generally acknowledged that occurrences of these phenomena are increasing throughout the world's oceans, the reasons for this apparent increase remain debated and include not only eutrophication but increased observation efforts in coastal zones of the world. There is a rapidly advancing monitoring effort resulting from the perception of increased impacts from these HABs, manifested as expanding routine coastal monitoring programs, rapid development and deployment of new detection methods for individual species, toxins, and toxicities, and expansion of coastal modeling activities towards observational forecasts of bloom landfall and eventually bloom prediction. Together, these many efforts will provide resource managers with the tools needed to develop effective strategies for the management and mitigation of HABs and their frequently devastating impacts on the coastal environment.

Environment↗

Three-dimensional biofilm model with individual cells and continuum EPS matrix.

An innovative type of biofilm model is derived by combining an individual description of microbial particles with a continuum representation of the biofilm matrix. This hybrid model retains the advantages of each approach, while providing a more realistic description of the temporal development of biofilm structure in two or three spatial dimensions. The general model derivation takes into account any possible number of soluble components. These are substrates and metabolic products, which diffuse and react in the biofilm within individual microbial cells. The cells grow, divide, and produce extracellular polymeric substances (EPS) in a multispecies model setting. The EPS matrix is described by a continuum representation as incompressible viscous fluid, which can expand and retract due to generation and consumption processes. The cells move due to a pushing mechanism between cells in colonies and by an advective mechanism supported by the EPS dynamics. Detachment of both cells and EPS follows a continuum approach, whereas cells attach in discrete events. Two case studies are presented for model illustration. Biofilm consolidation is explained by shrinking due to EPS and cell degradation processes. This mechanism describes formation of a denser layer of cells in the biofilm depth and occurrence of an irregularly shaped biofilm surface under nutrient limiting conditions. Micro-colony formation is investigated by growth of autotrophic microbial colonies in an EPS matrix produced by heterotrophic cells. Size and shape of colonies of ammonia and nitrite-oxidizing bacteria (NOB) are comparatively studied in a standard biofilm and in biofilms aerated from a membrane side.

Biofilms↗

Simultaneous carbon and nitrogen removal from high strength domestic wastewater in an aerobic RBC biofilm.

High strength domestic wastewater discharges after no/partial treatment through sewage treatment plants or septic tank seepage field systems have resulted in a large build-up of groundwater nitrates in Rajasthan, India. The groundwater table is very deep and nitrate concentrations of 500-750 mg/l (113-169 as NO3(-)-N) are commonly found. A novel biofilm in a 3-stage lab-scale rotating biological contactor (RBC) was developed by the incorporation of a sulphur oxidising bacterium Thiosphaera pantotropha which exhibited high simultaneous removal of carbon and nitrogen in fully aerobic conditions. T. pantotropha has been shown to be capable of simultaneous heterotrophic nitrification and aerobic denitrification thereby helping the steps of carbon oxidation, nitrification and denitrification to be carried out concurrently. The first stage having T. pantotropha dominated biofilm showed high carbon and NH4(+)-N removal rates of 8.7-25.9 g COD/m2 d and 0.81-1.85 g N/m2 d for the corresponding loadings of 10.0-32.0 g COD/m2 d and 1.0-3.35 g N/m2 d. The ratio of carbon removed to nitrogen removed was close to 12.0. The nitrification rate increased from 0.81 to 1.8 g N/m2 d with the increasing nitrogen loading rates despite a high simultaneous organic loading rate. However, it fell to 1.53 g N/m2 d at a high load of 3.35 g N/m2 d and 32 g COD/m2 d showing a possible inhibition of the process. A simultaneous 44-63% removal of nitrogen was also achieved without any significant NO2(-)-N or NO3(-)-N build-up. The second and third stages, almost devoid of any organic carbon, acted only as autotrophic nitrification units, converting the NH4(+)-N from stage 1 to nitrite and nitrate. Such a system would not need a separate carbon oxidation step to increase nitrification rates and no external carbon source for denitrification. The alkalinity compensation during denitrification for that destroyed in nitrification may also result in a high economy.

Biofilms↗

Deammonification in biofilm systems: population structure and function.

For the development of alternative concepts for the cost effective treatment of wastewaters with high ammonium content and low C/N-ratio, autotrophic consortia of micro-organisms with the ability to convert ammonium directly into N2 are of particular interest. Several full-scale industrial biofilm plants eliminating nitrogen without carbon source for years in a stable process, are suspected for some time to harbor active anaerobic ammonium oxidizers in deeper, oxygen-limited biofilm layers. In order to identify the processes of the single-stage nitrogen elimination (deammonification) in biofilm systems and to allocate them to the responsible micro-organisms, a deammonifying moving-bed pilot plant was investigated in detail. 15N-labelled tracer compounds were used as well as 16S rDNA libraries and in situ identification of dominant organisms. The usage of rRNA-targeted oligonucleotide probes (FISH) was particularly emphasized on the ammonium oxidizers of the beta-subclass of Proteobacteria and on the members of the order Planctomycetales. The combined application of these methods led to a deeper insight into the population structure and function of a deammonifying biofilm.

Biofilms↗