scholarly journals Monitoring Methanotrophic Bacteria in Hybrid Anaerobic-Aerobic Reactors with PCR and a Catabolic Gene Probe

1999 ◽  
Vol 65 (2) ◽  
pp. 381-388 ◽  
Author(s):  
Carlos B. Miguez ◽  
Chun F. Shen ◽  
Denis Bourque ◽  
Serge R. Guiot ◽  
Denis Groleau
Keyword(s):  
Pcr Amplification ◽  
Granular Sludge ◽  
Anaerobic Sludge ◽  
Initial Population ◽  
Gene Probe ◽  
Soluble Methane Monooxygenase ◽  
Chlorinated Pollutants ◽  
Pure Cultures ◽  
Amplification Assay ◽  
Aerobic Bioreactors

ABSTRACT We attempted to mimic in small upflow anaerobic sludge bed (UASB) bioreactors the metabolic association found in nature between methanogens and methanotrophs. UASB bioreactors were inoculated with pure cultures of methanotrophs, and the bioreactors were operated by using continuous low-level oxygenation in order to favor growth and/or survival of methanotrophs. Unlike the reactors in other similar studies, the hybrid anaerobic-aerobic bioreactors which we used were operated synchronously, not sequentially. Here, emphasis was placed on monitoring various methanotrophic populations by using classical methods and also a PCR amplification assay based on themmoX gene fragment of the soluble methane monooxygenase (sMMO). The following results were obtained: (i) under the conditions used, Methylosinus sporium appeared to survive better than Methylosinus trichosporium; (ii) the PCR method which we used could detect as few as about 2,000 sMMO gene-containing methanotrophs per g (wet weight) of granular sludge; (iii) inoculation of the bioreactors with pure cultures of methanotrophs contributed greatly to increases in the sMMO-containing population (although the sMMO-containing population decreased gradually with time, at the end of an experiment it was always at least 2 logs larger than the initial population before inoculation); (iv) in general, there was a good correlation between populations with the sMMO gene and populations that exhibited sMMO activity; and (v) inoculation with sMMO-positive cultures helped increase significantly the proportion of sMMO-positive methanotrophs in reactors, even after several weeks of operation under various regimes. At some point, anaerobic-aerobic bioreactors like those described here might be used for biodegradation of various chlorinated pollutants.

Study on performance of granular ANAMMOX process and characterization of the microbial community in sludge

2006 ◽  
Vol 54 (8) ◽  
pp. 197-207 ◽  
Author(s):  
Y. Yang ◽  
J. Zuo ◽  
Z. Quan ◽  
S. Lee ◽  
P. Shen ◽  
...  
Keyword(s):  
River Sediments ◽  
Pcr Amplification ◽  
Granular Sludge ◽  
Anaerobic Sludge ◽  
Anammox Bacteria ◽  
Formation Mechanisms ◽  
Ammonium Oxidation ◽  
Phylogenetic Characterization ◽  
Volumetric Loading

Anaerobic Ammonium Oxidation (ANAMMOX) is a novel biological nitrogen removal process, which is regarded as the most economical process at present. In this paper, two lab-scale UASB reactors, one of which was inoculated with the mixture of anaerobic sludge and aerobic sludge, the other with river sediments, were started up, using the inorganic synthetic water containing ammonium and nitrite as influent. After 421 days' and 356 days operation respectively, the ammonium removal efficiencies in two reactors reached 94% and 86% respectively, the total nitrogen volumetric loading rates were 2.5 and 1.6 kgN/m3.d. ANAMMOX granules were obtained in both reactors; the color of most granules was brown, but some of them were red. Based on the observation and studies on the microstructure of the granules, three kinds of ANAMMOX granular sludge formation mechanisms were proposed: adhering biofilm and disintegrated granular core mechanism, adhering biofilm and inorganic core mechanism and the self-coherence mechanism. For phylogenetic characterization of anaerobic ammonium oxidizers,16S rDNA approach was performed using Planctomycetales-specific PCR amplification. The dominant anammox bacteria occupied more than 90% of Planctomycetales-specific bacteria, and 27% of all bacteria in reactors. The dominant anammox bacteria distantly related to all currently reported candidate anammox genera. Functional gene of amoA was analyzed to investigate the ‘aerobic’ ammonium-oxidizing bacteria in β-Proteobacteria. The ‘aerobic’ ammonium-oxidizing bacteria were more diverse than anammox bacteria, but most of them clustered in anoxic ammonium-oxidizing Nitrosomonas eutropha/europaea groups. The composition of ‘aerobic’ ammonium-oxidizing bacteria is only 2% of all of bacteria in reactors.

Toxicity of Sulfite and Cadmium to Anaerobic Granular Sludge

1994 ◽  
Vol 29 (4) ◽  
pp. 581-598
Author(s):  
C.F. Shew ◽  
N. Kosaric
Keyword(s):  
Granular Sludge ◽  
Upflow Anaerobic Sludge Blanket ◽  
Shock Load ◽  
Anaerobic Sludge ◽  
Anaerobic Granular Sludge ◽  
Scanning Electron Micrographs ◽  
Bacterial Growth Rate ◽  
Digestion Rate ◽  
Sulfate Reducing ◽  
Load Conditions

Abstract Toxicity of sulfite (Na2SO3) and cadmium (CdCl2) ions to anaerobic granular sludge was investigated in 1.2 litre bench-scale upflow anaerobic sludge blanket (UASB) reactors during process acclimation and shock load conditions. Minimal sulfite toxicity was observed under gradual and shock load conditions at sulfite concentrations of up to 1000 mg S/L if proper acclimation was allowed to occur. No long-term toxic effects were observed although the COD digestion rate was temporarily inhibited by shock load of sulfite. Scanning electron micrographs indicated that more sulfate-reducing bacteria were present in the granules developed in the reactors with sulfite supplement although rod-shaped Methanosaeta-like bacteria were still dominant. High bacterial growth rate was observed in the reactors which were supplied with the feed containing sulfite. The COD digestion rate was inhibited at a cadmium loading rate of 2.4 g Cd per day under both acclimation and shock load conditions. Acclimation did not seem to improve the bacteria to tolerate the toxicity of cadmium. The concentration of free cadmium was very low in the reactors under normal conditions, but increased rapidly when the COD digestion in the reactors ceased. The bacteria could not be reactivated after inhibited by cadmium. When reactors were operated at low specific COD loading rates, more inorganic precipitates were formed inside the granules which consequently settled faster.

Anaerobic Thermophilic (55°C) Treatment of TMP Whitewater in Reactors Based on Biomass Attachment and Entrapment

1999 ◽  
Vol 40 (11-12) ◽  
pp. 67-75 ◽  
Author(s):  
Sigrun J. Jahren ◽  
Jukka A. Rintala ◽  
Hallvard Ødegaard
Keyword(s):  
Granular Sludge ◽  
Biofilm Reactor ◽  
Upflow Anaerobic Sludge Blanket ◽  
Hybrid Reactor ◽  
Anaerobic Sludge ◽  
Loading Rates ◽  
Anaerobic Reactors ◽  
Multi Stage ◽  
Degradation Rates ◽  
Thermomechanical Pulping

Thermomechanical pulping (TMP) whitewater was treated in thermophilic (55°C) anaerobic laboratory-scale reactors using three different reactor configurations. In all reactors up to 70% COD removals were achieved. The anaerobic hybrid reactor, composed of an upflow anaerobic sludge blanket (UASB) and a filter, gave degradation rates up to 10 kg COD/m3d at loading rates of 15 kg COD/m3d and hydraulic retention time (HRT) of 3.1 hours. The anaerobic multi-stage reactor, consisting of three compartments, each packed with granular sludge and carrier elements, gave degradation rates up to 9 kg COD/m3d at loading rates of 15-16 kg COD/m3d, and HRT down to 2.6 hours. Clogging and short circuiting eventually became a problem in the multi-stage reactor, probably caused by too high packing of the carriers. The anaerobic moving bed biofilm reactor performed similar to the other reactors at loading rates below 1.4 kg COD/m3d, which was the highest loading rate applied. The use of carriers in the anaerobic reactors allowed short HRT with good treatment efficiencies for TMP whitewater.

The effect of extreme temperatures (70–80°C) on the effluent quality and sludge characteristics of UASB reactors

1997 ◽  
Vol 36 (6-7) ◽  
pp. 325-332 ◽  
Author(s):  
Raghida Lepistö ◽  
Jukka Rintala
Keyword(s):  
Volatile Fatty Acids ◽  
Granular Sludge ◽  
Anaerobic Sludge ◽  
Methanogenic Activity ◽  
Sludge Characteristics ◽  
Effluent Quality ◽  
Start Up ◽  
Thermomechanical Pulping ◽  
Increasing Temperature ◽  
Uasb Reactors

The study focused on the effluent quality and sludge characteristics during the start-up and operation of extreme thermophilic (70 to 80°C) upflow anaerobic sludge bed (UASB) reactors, inoculated with mesophilic and thermophilic granular sludge and fed with acetate, volatile fatty acids (VFA), and thermomechanical pulping (TMP) whitewater. Low effluent quality and long start-up periods were observed during the start-up of the 70 to 76°C, VFA-fed UASB reactors inoculated with mesophilic granulae, while better effluent quality and considerably shorter start-up periods were observed when thermophilic (55/70°C) inocula were used. With VFA feed, a significant amount of acetate was removed at 70°C and even at 80°C, while propionate removal was negligible. With TMP whitewater feed, low VFA effluent concentration was obtained at 70°C. The volatile solids (VS) and the VS/total solids (TS) content of the sludge decreased significantly during the first 2–3 months of operation when mesophilic inocula were used. The initial specific methanogenic activity (ISMA) of the extreme thermophilic sludge decreased with increasing temperature and was slightly higher on glucose than on acetate. At 70 to 80°C, various rod-like bacteria were dispersed through the granulae in either individual or in low density micro colonies surrounded with a varying degree of precipitates.

scholarly journals Microbial Diversity Dynamics in a Methanogenic-Sulfidogenic UASB Reactor

2021 ◽  
Vol 18 (3) ◽  
pp. 1305
Author(s):  
E. Fernández-Palacios ◽  
Xudong Zhou ◽  
Mabel Mora ◽  
David Gabriel
Keyword(s):  
Paper Industry ◽  
Chemical Species ◽  
Methanogenic Archaea ◽  
Granular Sludge ◽  
Upflow Anaerobic Sludge Blanket ◽  
Uasb Reactor ◽  
Anaerobic Sludge ◽  
Rrna Gene ◽  
Sulfate Reducing ◽  
Long Run

In this study, the long-term performance and microbial dynamics of an Upflow Anaerobic Sludge Blanket (UASB) reactor targeting sulfate reduction in a SOx emissions treatment system were assessed using crude glycerol as organic carbon source and electron donor under constant S and C loading rates. The reactor was inoculated with granular sludge obtained from a pulp and paper industry and fed at a constant inlet sulfate concentration of 250 mg S-SO42−L−1 and a constant C/S ratio of 1.5 ± 0.3 g Cg−1 S for over 500 days. Apart from the regular analysis of chemical species, Illumina analyses of the 16S rRNA gene were used to study the dynamics of the bacterial community along with the whole operation. The reactor was sampled along the operation to monitor its diversity and the changes in targeted species to gain insight into the performance of the sulfidogenic UASB. Moreover, studies on the stratification of the sludge bed were performed by sampling at different reactor heights. Shifts in the UASB performance correlated well with the main shifts in microbial communities of interest. A progressive loss of the methanogenic capacity towards a fully sulfidogenic UASB was explained by a progressive wash-out of methanogenic Archaea, which were outcompeted by sulfate-reducing bacteria. Desulfovibrio was found as the main sulfate-reducing genus in the reactor along time. A progressive reduction in the sulfidogenic capacity of the UASB was found in the long run due to the accumulation of a slime-like substance in the UASB.

Intercistronic heterogeneity of the 16S–23S rRNA spacer region among Pseudomonas strains isolated from subterranean seeds of hog peanut (Amphicarpa bracteata)

Microbiology ◽  
2009 ◽  
Vol 155 (8) ◽  
pp. 2630-2640 ◽  
Author(s):  
J. T. Tambong ◽  
R. Xu ◽  
E. S. P. Bromfield
Keyword(s):  
Root Zone ◽  
Phylogenetic Analyses ◽  
Melting Curve ◽  
Pcr Amplification ◽  
23S Rrna ◽  
Melting Curve Analysis ◽  
Its1 Region ◽  
A Genome ◽  
Pure Cultures ◽  
Intercalating Dye

Intercistronic heterogeneity of the 16S–23S rRNA internal transcribed spacer regions (ITS1) was investigated in 29 strains of fluorescent pseudomonads isolated from subterranean seeds of Amphicarpa bracteata (hog peanut). PCR amplification of the ITS1 region generated one or two products from the strains. Sequence analysis of the amplified fragments revealed an ITS1 fragment of about 517 bp that contained genes for tRNAIle and tRNAAla in all 29 strains; an additional smaller ITS1 of 279 bp without tRNA features was detected in 15 of the strains. The length difference appeared to be due to deletions of several nucleotide blocks between the 70 bp and 359 bp positions of the alignment. The end of the deletions in the variant ITS1 type coincided with the start of antiterminator box A, which is homologous to box A of other bacteria. Phylogenetic analyses using the neighbour-joining algorithm revealed two major phylogenetic clusters, one for each of the ITS1 types. Using a single specific primer set and the DNA-intercalating dye SYBR Green I for real-time PCR and melting-curve analysis produced highly informative curves with one or two recognizable melting peaks that readily distinguished between the two ITS1 types in pure cultures. The assay was used to confirm the presence of the variant ITS1 type in the Pseudomonas community in total DNA from root-zone soil and seed coats of hog peanut. Heterogeneity of the ITS1 region between species has potential for studying molecular systematics and population genetics of the genus Pseudomonas, but the presence of non-identical rRNA operons within a genome may pose problems.

Improved nitrogen removal in upflow anaerobic sludge blanket (UASB) reactors by incorporation of Anammox bacteria into the granular sludge

2004 ◽  
Vol 49 (11-12) ◽  
pp. 69-76 ◽  
Author(s):  
J.E. Schmidt ◽  
D.J. Batstone ◽  
I. Angelidaki
Keyword(s):  
In Situ Hybridisation ◽  
Granular Sludge ◽  
Ammonia Removal ◽  
Appropriate Technology ◽  
Upflow Anaerobic Sludge Blanket ◽  
Anaerobic Sludge ◽  
The Third ◽  
Anammox Activity ◽  
Anaerobic Sludge Blanket ◽  
Uasb Reactors

Upflow anaerobic sludge blanket reactors may offer a number of advantages over conventional mixed-tank, SBR, and biofilm reactors, including high space-loading, low footprint, and resistance to shocks and toxins. In this study, we assessed the use of upflow anaerobic sludge blanket (UASB) reactor technology as applied to anaerobic ammonia removal, or Anammox. Four 200 ml UASB reactors were inoculated with 50% (by volume) anaerobic granular sludge and 50% flocular sludge from different sources (all with the potential for containing Anammox organisms). Tools used to assess the reactors included basic analyses, fluorescent in-situ hybridisation, and mathematical modelling, with statistical non-linear parameter estimation. Two of the reactors showed statistically identical Anammox activity (i.e., identical kinetic parameters), with good ammonia and nitrite removal (0.14 kgNHx m-3 reactor day-1, with 99% ammonia removal). The third reactor also demonstrated significant Anammox activity, but with poor identifiability of parameters. The fourth reactor had no statistical Anammox activity. Modelling indicated that poor identifiability and performance in the third and fourth reactors were related to an excess of reduced carbon, probably originating in the inoculum. Accumulation of Anammox organisms was confirmed both by a volume loading much lower than the growth rate, and response to a probe specific for organisms previously reported to mediate Anammox processes. Overall, the UASB reactors were effective as Anammox systems, and identifiability of the systems was good, and repeatable (even compared to a previous study in a rotating biological contactor). This indicates that operation, design, and analysis of Anammox UASB reactors specifically, and Anammox systems in general, are reliable and portable, and that UASB systems are an appropriate technology for this process.

The biobed® EGSB (expanded granular sludge bed) system covers shortcomings of the upflow anaerobic sludge blanket reactor in the chemical industry

1997 ◽  
Vol 35 (10) ◽  
pp. 183-188 ◽  
Author(s):  
George R. Zoutberg ◽  
Peter de Been
Keyword(s):  
Waste Water ◽  
Chemical Industry ◽  
Granular Sludge ◽  
Upflow Anaerobic Sludge Blanket ◽  
Anaerobic Sludge ◽  
Special Configuration ◽  
Anaerobic Reactor ◽  
High Concentrations ◽  
Anaerobic Sludge Blanket ◽  
Brewery Yeast

In this paper a new type of anaerobic reactor is presented. The system has been developed by Biothane Systems and is marketed under the name Biobed® EGSB reactor (Expanded Granular Sludge Bed). In this reactor it is possible to grow and maintain a granular sludge under high liquid (10 m/h) and gas velocities (7 m/h). The most striking feature is the growth of biomass in a granular form, similar to the UASB granules: no carrier material is used. The process is specially suitable to treat waste water that contains compounds that are toxic in high concentrations and that only can be degraded in low concentrations (chemical industry). An example is given for a waste water originating from a chemical factory (Caldic Europoort) in the Netherlands. In this factory formaldehyde is produced from methanol. The waste water is characterised by high concentrations of these compounds (formaldehyde to 10 g/l and methanol to 20 g/l). Due to the special configuration of the anaerobic reactor it is possible to realise a removal efficiency for both compounds of more than 98%. It is also possible to operate the reactor as an ultra high loaded anaerobic reactor (to 30 kg COD/m3.day) for applications in other sectors of industry (e.g. brewery, yeast, sugar, corn ethanol production etc).

Sign in / Sign up

Export Citation Format

Share Document