Water 2016, 8, 172
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Supplementary Materials: Effects of Wash-Out
Dynamics on Nitrifying Bacteria in Aerobic Granular
Sludge During Start-Up at Gradually Decreased
Settling Time
Enikö Szabó, Malte Hermansson, Oskar Modin, Frank Persson and Britt-Marie Wilén
Figure S1. Micrographs of the sludge in R1 (A) and R2 (B) taken 10 days after start-up.
Water 2016, 8, 172; doi:10.3390/w8050172
www.mdpi.com/journal/water
Water 2016, 8, 172
Figure S2. Micrographs of the sludge in R1 (A) and R2 (B) taken 28 days after start-up.
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Figure S3. Results of the qPCR analyses. Gene copy numbers of AOB amoA (a); and Universal 16S
rRNA (b).
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Figure S4. Relative abundance of TRFs. Samples are marked with the identifier of the reactor (R1 or R2) and the number of days since start-up. SS = seed sludge.
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Figure S5. Effluent TSS concentration in the reactors.
Figure S6. The impact of settling time on the SRT during start-up. The changes in settling time (min)
are indicated with arrows and Arabic numerals.
Figure S7. Variation of the pH in the reactors (measured at the end of the cycles).
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Figure S8. Typical pattern of pH variation during a cycle in R1 (a) and R2 (b). The gray line marks
the five minute anaerobic filling, i.e., the beginning of the cycle.
Table S1. Primer pairs and annealing temperatures used for PCR and qPCR analysis.
Primer Pairs
amoA1F
amoA2R
CTO189A/B/C
CTO654
FGPS872
FGPS1269
Nsr1113F
Nsr1264R
Uni1055F
Uni1392R
Target
Organism
AOB (Betaproteobacteria)
AOB (Betaproteobacteria)
Application
Annealing
Temp. (°C)
Detection
PCR and qPCR
54
+
PCR and qPCR
57
+
Degrange and Bardin
1995
Nitrobacter sp.
PCR
50
–
Dionisi et al. 2002
Nitrospira sp.
PCR and qPCR
65
+
Ferris et al. 1996
Most bacteria
PCR and qPCR
54
+
Reference
Rotthauwe et al. 1997
Kowalchuk et al. 1997
References
1.
2.
Degrange, V; Bardin, R. Detection and counting of Nitrobacter populations in soil by PCR. Appl.
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Dionisi, H.M.; Layton, A.C.; Harms, G.; Igrid, R.; Robinson, K.G.; Sayler, G.S. Quantification of
Nitrosomonas oligotropha -Like Ammonia-Oxidizing Bacteria and Nitrospira spp. from Full-Scale
Wastewater Treatment Plants by Competitive PCR. Appl. Environ. Microbiol. 2002, 68, 245–253.
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Ferris, M.J.; Muyzer, G.; Ward, D.M. Denaturing gradient gel electrophoresis profiles of 16S rRNAdefined populations inhabiting a hot spring microbial mat community. Appl. Environ. Microbiol. 1996,
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