Table 1.

Average acceptor and donor fluxes for each stage of the MBfR experiments.

NO3--N
Electron donor
consumed
Flux
2
b
(g N/m -d)
(mmol
CH4/m2-d)
NA
0
NA
0
0.08±0.007
6.4±0.6
0.55±0.09
47.4±7.2
0.30±0.003
26.0±0.3
NA
0
NA
0

stagesa
1
2
3
4
5
6
7

NO2--N
Electron donor
Flux
consumed
2
(g N/m -d)
(mmol
CH4/m2-d)
0.10±0.001
5.97±0.06
NA
0
NA
0
NA
0
NA
0
NA
0
0.39±0.01
23.2±0.6

ClO4Electron donor
Flux
consumed
2
(g/m -d)
(mmol
CH4/m2-d)
0.02±0.002
0.29±0.03
0.05±0.006
0.82±0.09
0.06±0.001
0.96±0.01
0.002±0.0004
0.03±0.006
0.07±0.006
1.03±0.09
0.39±0.006
5.91±0.09
0.19±0.03
2.87±0.44

Electron donor (CH4)
Maximum CH4
Actual CH4 flux
flux
(mmol
(mmol
2
b
CH
4/m -d)
CH4/m2-d)b, c
57.9
6.26±0.06
57.9
0.82±0.09
57.9
7.35±0.60
57.9
47.5±7.20
86.8
27.0±0.26
86.8
5.91±0.09
86.8
26.0±0.59

NA = not applicable
a:

Gas pressure was 10 psi for stage 1-4, 15 psi for stage 5-7.

b:

Calculated from equations 2 through 4.

c:

Maximum CH4

flux was calculated by Tang et al (2012), considering the Pm-if (CH4 pressure at the interface of membrane and liquid film) was 0, and
a theoretical maximum CH4 flux can be obtained in the liquid phase.

c: 1 bar= 14.5 psi

Figure 1.

Headspace CH4 pressure during the CH4-permeation experiment.

Steady state Phs

was achieved at ~15 hours.

2

Figure 2.

(A) NO2-, NO3-, and ClO4- concentrations in the MBfR influent and effluent.

(B)

NO2-, NO3-, and ClO4- removal percentages.

3

Figure 3.

The average fluxes of NO3-, NO2-, and ClO4- along with the qPCR-based abundances

of functional genes through all stages.

The narG gene copy number in Stage 4 is plotted as

open black triangle so that it can been seen within the green bar, while narG gene copy number
in other stages are plotted as solid green triangles. Absence of bars indicates that the error is
smaller than the symbol size.

4

Figure 4.

Linear regression of mcrA, pMMO, and mcrA+pMMO functional genes against the

sum of narG, nirS, and pcrA genes.

5

Figure 5.

The potential ClO4- reduction pathways by the ANMO-D culture.

A:

ANMO-PR,

Anaerobic methane oxidation coupled to perchlorate reduction, carried out by one bacterium
that dismutates ClO2- to form Cl- and O2 intracellularly, with the O2 used as a co-substrate for
methane mono-oxygenation by itself (Intra-Aerobic Type);

B:

mAMO-PR:

micro-Aerobic

methane oxidation coupled to perchlorate reduction, carried out by two microorganisms:
bacteria reduce ClO4- to Cl- and produce O2 extracellularly, and the O2 is utilized by
methanotrophs to oxidize methane (Extracellular micro-Aerobic Type).

6