Use of microautoradiography and fluorescent in situ hybridization for characterization of microbial activity in activated sludge

Nielsen, Per Halkjær; Andreasen, Kjær; Lee, N.; Wagner, Michael

doi:10.2166/wst.1999.0001

Cited by 30 publications

(28 citation statements)

References 26 publications

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“…For instance, the small filamentous organisms are morphologically similar to both the photoautotrophic LPP seen to dominate in Salt Pond after Hurricane Frances (Yannarell et al , 2007) and to the photoorganotrophic Chloroflexus shown to dominate in a similar hypersaline microbial mat (Nübel et al , 2001; Ley et al , 2006). Further molecular characterization of the Salt Pond microbial community will be useful in the development of molecular probes with high taxonomic resolution, so that techniques such as combined microautoradiography and fluorescent in situ hybridization (Nielsen et al , 1999) may help clarify the role of specific organisms involved in DOM utilization in this system.…”

Section: Discussionmentioning

confidence: 99%

Effects of salinity and light on organic carbon and nitrogen uptake in a hypersaline microbial mat

Yannarell

Paerl

2007

FEMS Microbiology Ecology

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Utilization of dissolved organic matter (DOM) is thought to be the purview of heterotrophic microorganisms, but photoautotrophs can take up dissolved organic nitrogen (DON) and dissolved organic carbon (DOC). This study investigated DOC and DON uptake in a laminated cyanobacterial mat community from hypersaline Salt Pond (San Salvador, Bahamas). The total community uptake of (3)H-labeled substrates was measured in the light and in the dark and under conditions of high and low salinity. Salinity was the primary control of DOM uptake, with increased uptake occurring under low-salinity, 'freshened' conditions. DOC uptake was also enhanced in the light as compared with the dark and in samples incubated with the photosystem II inhibitor 3(3,4-dichlorophenyl)-1, 1-dimethylurea, suggesting a positive association between photosynthetic activity and DOC uptake. Microautoradiography revealed that some DOM uptake was attributed to cyanobacteria. Cyanobacteria DOM uptake was negatively correlated with that of smaller filamentous microorganisms, and DOM uptake by individual coccoid cells was negatively correlated with uptake by colonial coccoids. These patterns of activity suggest that Salt Pond microorganisms are engaged in resource partitioning, and DOM utilization may provide a metabolic boost to both heterotrophs and photoautrophs during periods of lowered salinity.

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Section: Discussionmentioning

confidence: 99%

Effects of salinity and light on organic carbon and nitrogen uptake in a hypersaline microbial mat

Yannarell

Paerl

2007

FEMS Microbiology Ecology

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“…Substrate uptake patterns for S. piniformis were determined under in situ conditions by MAR as described by Nielsen and colleagues (1999). Two ml samples of diluted activated sludge foam adjusted to a final concentration of 1gSS l −1 were incubated with the following substrates: acetate, glucose, glycine, glycerol, oleic acid, palmitic acid, benzoic acid, cholesterol and estradiol.…”

Section: Methodsmentioning

confidence: 99%

The in situ physiology of Skermania piniformis in foams in Australian activated sludge plants

Eales

Nielsen

Seviour

et al. 2006

Environmental Microbiology

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The in situ physiology of the filamentous bacterium Skermania piniformis frequently seen in activated sludge foams in Australia was investigated. An oligonucleotide probe, Spin1449, targeting the 16S rRNA of S. piniformis was designed for its identification by fluorescence in situ hybridization (FISH), validated with pure cultures and applied successfully to foam samples from two geographically distant Australian plants. While filaments of this bacterium appeared to be comparatively hydrophobic, the organism had no clear preference for hydrophobic or hydrophilic substrates. In both foams examined using microautoradiography (MAR), filaments selectively took up substrates under aerobic and anoxic (NO(3) (-)) but not anaerobic or anoxic (NO(2) (-)) conditions. Skermania piniformis assimilated oleic acid, palmitic acid, glycerol and glycine. Ectoenzyme activities detected suggest that S. piniformis has an ability to assimilate a greater range of substrates than might be concluded from the MAR data obtained here. Based on the substrate uptake data presented here, an anaerobic selector may work for controlling S. piniformis in activated sludge systems.

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“…This experimental technique links FISH, used to identify individual species within a MMC through fluorescently labeled ribosomal RNA‐targeted oligonucleotide probes (DeLong et al, 1989), with MAR to detect substrate specific consumption using radioactively labeled substrates. MAR‐FISH is becoming a widely used tool to study the ecophysiology of microbial communities (Albuquerque et al, 2013; Hagman et al, 2008; Nielsen et al, 1999). The concurrent application of MAR and FISH techniques to polyhydroxyalkanoates (PHA) producing MMCs was reported, for the first time, by Albuquerque et al (2013).…”

Section: Introductionmentioning

confidence: 99%

Segregated flux balance analysis constrained by population structure/function data: The case of PHA production by mixed microbial cultures

Pardelha

Albuquerque

Carvalho

et al. 2013

Biotech & Bioengineering

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In this study we developed a segregated flux balance analysis (FBA) method to calculate metabolic flux distributions of the individual populations present in a mixed microbial culture (MMC). Population specific flux data constraints were derived from the raw data typically obtained by the fluorescence in situ hybridization (FISH) and microautoradiography (MAR)-FISH techniques. This method was applied to study the metabolic heterogeneity of a MMC that produces polyhydroxyalkanoates (PHA) from fermented sugar cane molasses. Three populations were identified by FISH, namely Paracoccus sp., Thauera sp., and Azoarcus sp. The segregated FBA method predicts a flux distribution for each of the identified populations. The method is shown to predict with high accuracy the average PHA storage flux and the respective monomeric composition for 16 independent experiments. Moreover, flux predictions by segregated FBA were slightly better than those obtained by nonsegregated FBA, and also highly concordant with metabolic flux analysis (MFA) estimated fluxes. The segregated FBA method can be of high value to assess metabolic heterogeneity in MMC systems and to derive more efficient eco-engineering strategies. For the case of PHA-producing MMC considered in this work, it becomes apparent that the PHA average monomeric composition might be controlled not only by the volatile fatty acids (VFA) feeding profile but also by the population composition present in the MMC.

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Use of microautoradiography and fluorescent in situ hybridization for characterization of microbial activity in activated sludge

Cited by 30 publications

References 26 publications

Effects of salinity and light on organic carbon and nitrogen uptake in a hypersaline microbial mat

Effects of salinity and light on organic carbon and nitrogen uptake in a hypersaline microbial mat

The in situ physiology of Skermania piniformis in foams in Australian activated sludge plants

Segregated flux balance analysis constrained by population structure/function data: The case of PHA production by mixed microbial cultures

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