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Diversity and enrichment of nitrite-dependent anaerobic methane oxidizing bacteria from wastewater sludge.

Luesken FA, van Alen TA, van der Biezen E, Frijters C, Toonen G, Kampman C, Hendrickx TL, Zeeman G, Temmink H, Strous M, Op den Camp HJ, Jetten MS - Appl. Microbiol. Biotechnol. (2011)

Bottom Line: This enrichment was monitored using specific pmoA primers and M. oxyfera cells were visualized with fluorescence oligonucleotide probes.After 112 days, the enrichment consumed up to 0.4 mM NO(2)(-) per day.The results of this study show that appropriate sources of biomass, enrichment strategies, and diagnostic tools existed to start and monitor pilot scale tests for the implementation of nitrite-dependent methane oxidation in wastewater treatment at ambient temperature.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Institute for Water and Wetland Research, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.

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Increasing population of M. oxyfera-type bacteria in Lieshout enrichment culture, visualized with FISH. The cells were hybridized with the probes S-*-DBACT-1027-a-A-18 specific for the NC10 phylum (red) and S-D-Bact-0338-a-A-18 that target most, but not all bacteria (dark blue). a There were no M. oxyfera cells detected in the inoculum, using FISH. b After an incubation period of 64 days in a 3-l reactor, M. oxyfera-type bacteria were enriched for ~2–3% (represented in pink, caused by double hybridization of DBACT1027 and EUB338). c Clusters and some detached cells of M. oxyfera-type bacteria are present in the culture after 308 days (represented in pink). M. oxyfera-type bacteria were enriched for ~60–70%. Probe S-*-DBACT-0193-a-A-18 specific for M. oxyfera bacteria showed the same result. Scale bar is 20 μm
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Fig3: Increasing population of M. oxyfera-type bacteria in Lieshout enrichment culture, visualized with FISH. The cells were hybridized with the probes S-*-DBACT-1027-a-A-18 specific for the NC10 phylum (red) and S-D-Bact-0338-a-A-18 that target most, but not all bacteria (dark blue). a There were no M. oxyfera cells detected in the inoculum, using FISH. b After an incubation period of 64 days in a 3-l reactor, M. oxyfera-type bacteria were enriched for ~2–3% (represented in pink, caused by double hybridization of DBACT1027 and EUB338). c Clusters and some detached cells of M. oxyfera-type bacteria are present in the culture after 308 days (represented in pink). M. oxyfera-type bacteria were enriched for ~60–70%. Probe S-*-DBACT-0193-a-A-18 specific for M. oxyfera bacteria showed the same result. Scale bar is 20 μm

Mentions: In order to get an impression of the relative abundance of M. oxyfera cells compared to the other community members, fluorescence in situ hybridization (FISH) was performed using general and specific probes (Fig. 3). No M. oxyfera-type bacteria could be detected in the inoculum (Fig. 3a), probably because the number of cells was below detection level (Amann et al. 1995). This is consistent with the 16S rRNA and pmoA phylogenetic analysis (see above). Although PCR is a more sensitive technique compared to FISH, a nested PCR approach was necessary to detect M. oxyfera-type bacteria in the inoculum. After 64 days of cultivation in the 3-l bioreactor, approximately 2–3% of the microbial community hybridized with specific probes for M. oxyfera (Fig. 3b). After 308 days, the M. oxyfera population was enriched for approximately 60–70% (Fig. 3c). The enriched bacteria appeared to grow mainly in clusters, but some single cells were also visible as observed previously (Ettwig et al. 2009).Fig. 3


Diversity and enrichment of nitrite-dependent anaerobic methane oxidizing bacteria from wastewater sludge.

Luesken FA, van Alen TA, van der Biezen E, Frijters C, Toonen G, Kampman C, Hendrickx TL, Zeeman G, Temmink H, Strous M, Op den Camp HJ, Jetten MS - Appl. Microbiol. Biotechnol. (2011)

Increasing population of M. oxyfera-type bacteria in Lieshout enrichment culture, visualized with FISH. The cells were hybridized with the probes S-*-DBACT-1027-a-A-18 specific for the NC10 phylum (red) and S-D-Bact-0338-a-A-18 that target most, but not all bacteria (dark blue). a There were no M. oxyfera cells detected in the inoculum, using FISH. b After an incubation period of 64 days in a 3-l reactor, M. oxyfera-type bacteria were enriched for ~2–3% (represented in pink, caused by double hybridization of DBACT1027 and EUB338). c Clusters and some detached cells of M. oxyfera-type bacteria are present in the culture after 308 days (represented in pink). M. oxyfera-type bacteria were enriched for ~60–70%. Probe S-*-DBACT-0193-a-A-18 specific for M. oxyfera bacteria showed the same result. Scale bar is 20 μm
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3198195&req=5

Fig3: Increasing population of M. oxyfera-type bacteria in Lieshout enrichment culture, visualized with FISH. The cells were hybridized with the probes S-*-DBACT-1027-a-A-18 specific for the NC10 phylum (red) and S-D-Bact-0338-a-A-18 that target most, but not all bacteria (dark blue). a There were no M. oxyfera cells detected in the inoculum, using FISH. b After an incubation period of 64 days in a 3-l reactor, M. oxyfera-type bacteria were enriched for ~2–3% (represented in pink, caused by double hybridization of DBACT1027 and EUB338). c Clusters and some detached cells of M. oxyfera-type bacteria are present in the culture after 308 days (represented in pink). M. oxyfera-type bacteria were enriched for ~60–70%. Probe S-*-DBACT-0193-a-A-18 specific for M. oxyfera bacteria showed the same result. Scale bar is 20 μm
Mentions: In order to get an impression of the relative abundance of M. oxyfera cells compared to the other community members, fluorescence in situ hybridization (FISH) was performed using general and specific probes (Fig. 3). No M. oxyfera-type bacteria could be detected in the inoculum (Fig. 3a), probably because the number of cells was below detection level (Amann et al. 1995). This is consistent with the 16S rRNA and pmoA phylogenetic analysis (see above). Although PCR is a more sensitive technique compared to FISH, a nested PCR approach was necessary to detect M. oxyfera-type bacteria in the inoculum. After 64 days of cultivation in the 3-l bioreactor, approximately 2–3% of the microbial community hybridized with specific probes for M. oxyfera (Fig. 3b). After 308 days, the M. oxyfera population was enriched for approximately 60–70% (Fig. 3c). The enriched bacteria appeared to grow mainly in clusters, but some single cells were also visible as observed previously (Ettwig et al. 2009).Fig. 3

Bottom Line: This enrichment was monitored using specific pmoA primers and M. oxyfera cells were visualized with fluorescence oligonucleotide probes.After 112 days, the enrichment consumed up to 0.4 mM NO(2)(-) per day.The results of this study show that appropriate sources of biomass, enrichment strategies, and diagnostic tools existed to start and monitor pilot scale tests for the implementation of nitrite-dependent methane oxidation in wastewater treatment at ambient temperature.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Institute for Water and Wetland Research, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.

Show MeSH