Table S1.PCR amplifying conditions and sequencing data
Sample ID
PCR amplifying
Target microbe
hypervariable regions of 16s
rDNA
forward primer
reverse primer
PCR conditions
FS
DS
AD1
AD2
AD2
AD2
bacteria
V3V4
bacteria
V3V4
bacteria
V3V4
bacteria
V3V4
archaea
V5V6
fungi
ITS
338F
338F
338F
338F
787F
ITS1F
806R
806R
806R
806R
1059R
ITS2R
one pre-denaturation cycle at 98℃ for 5 min, 25 or 27
cycles of denaturation at 98℃ for 30 s, annealing at 50℃
for 30 s, and elongation at 72℃for 30 s, and one postelongation cycle at 72℃ for 5 min
Sequence of target microorganism
Valid Sequence number
20263
21462
23397
40372
29477
47775
High quality Sequence number
15172
16660
21697
28220
22992
45797
High quality sequence length
424454 bp
424454 bp
424454 bp
424454 bp
246253 bp
160293 bp
Phylum
2993
2065
1762
4394
42
323
Class
2992
2060
1728
4315
42
323
Order
2955
2027
181
3987
40
323
Family
2781
1870
1183
2969
38
323
Genus
2251
1388
597
1487
34
323
Species
1193
691
58
94
0
323
151
1
10
0
0
OTU of target microorganism
Unclassified
131
Species numbers of target microorganism
Phylum
42
42
44
51
2
5
Class
94
94
98
117
5
19
Order
142
142
136
179
6
42
Family
157
157
163
203
12
67
Genus
177
177
200
245
20
82
338F： 5-ACTCCTACGGGAGGCAGCAG-3 806R ： 5-GGACTACVSGGGTATCTAAT-3
787F：5- ATTAGATACCCSBGTAGTCC-3 1059R：5- GCCATGCACCWCCTCT-3
ITS1F：5- CTTGGTCATTTAGAGGAAGTAA-3 ITS2R： 5-GCTGCGTTCTTCATCGATGC-3
Table S2 Heavy metal contents of digested sludge
Heavy contents of digested
sludge
Total mercury(Hg)
Total cadmium(Cd)
Total chromium(Cr)
Total copper(Cu)
Total lead(Pb)
Units
mg/kg dry
sludge
mg/kg dry
sludge
mg/kg dry
sludge
mg/kg dry
sludge
mg/kg dry
Numbe
r
10
Min
Max
mean
0.22
9.27
5.20
IC50
(mg/L)
-
10
0.48
7.79
3.28
36-330
10
30.1
211.0
97.8
27-250
10
3.23
160.00
114.31
130
10
12.2
1156.0
56.7
27.7
sludge
mg/kg dry
sludge
mg/kg dry
sludge
mg/kg dry
sludge
Total zinc(Zn)
Total nickel(Ni)
Total arsenic(As)
10
1660
1010
624
7.5-270
10
7.46
361.00
134.38
-
10
4.05
31.30
17.16
IC50：half maximal inhibitory concentration(Chen et al.,2014)
Table S3 Biogas contents[5]
Biogas contents
CH4
CO2
O2
N2
H2
CO
H2S
Units
Number
Min
Max
mean
Vol.%
Vol.%
Vol.%
Vol.%
Vol.%
Vol.%
Vol.%
24
24
24
24
24
24
24
62.7
12.1
0.1
0.18
≤0.0075
≤0.007
0.23
69.1
31.2
4.57
7.75
0.14
≤0.007
0.61
65.3
26.2
0.96
1.73
≤0.0075
≤0.007
0.40
Optimum
values
65-70
30-35-
Extreme
values
60-75
25-40
-
Table S4. Summary of dominant bacteria identified in this study and previous studies
Phylogenetic
group
Average proportion of bacterial groups at the phylum level of ADS
Lee et al.,
2012
Sundberg et
al.,2013
Guo et al.,
2015
Yang, et
al.,2014
Proteobacteria
7.2-33.8%
4-6%
41.2%
24.3-25.7%
Bacteroidetes
10.2-29.2%
14-15%
9.6%
Firmicutes
8.0-27.6%
25-69%
12.5%
9.52–
13.50 %
7.18–
10.65 %
7.53–9.46 %
Chloroflexi
0.3-9.3%
0.5-2%
N.A.
1.75–4.54 %
7.2-9.8%
WWE1
N.A.
N.A.
N.A.
N.A.
0-1.3%
Actinobacteria
0.3-3.5%
4-6%
5.2%
5.05–7.89 %
3.0-3.2%
Synergistetes
0.6-3.2%
N.A.
N.A.
N.A.
2.6-3.1%
Spirochaetes
0.1-2.3%
4-6%
N.A.
1.27–3.42 %
2.5-6.5%
Thermotogae
0.1-4.1%
N.A.
N.A.
2.42–6.18 %
0.8-2.6%
Pyrosequencing
Platform
454
454
Illumina
Illumina
Illumina
V1,V2,V3
V3,V4
All regions
All regions
V3,V4
Hypervariable
regions of 16S
rDNA
amplification and
sequencing
This study
23.4-23.6%
16.6-17.0%
Classification
Database
RDP
RDP
Feedstock of
anaerobic
digesters
SS, SS
+food waste
recycling
wastewater
SS,
codigesting
various
combinations
Entire
available
source
databases in
MG-RAST
SILVA SSU
database
Greengenes
SS
SS
SS
Fig. S5 MEGAN profile showing the bacterial diversity for AD1 andAD2 at the genus level
Fig. S6 Bar charts showing the major genes in the dominant bacteria of AD1 and AD2 sludge for
“amino acid metabolism” , “carbohydrate metabolism” and “energy metabolism pathways”
Fig. S7 Bar charts showing the major genes in the dominant bacteria of AD1 and FS sludge for
“carbohydrate metabolism”, “xenobiotics biodegradation and metabolism” and “metabolism of other
amino acids” pathways
Fig. S8 Heat map comparing the bacterial diversities of DS, AD1 and FS at genus level
Table S9 Pathogen genera of DS and FS
Taxon
Aeromonas
Bacillus
Brevibacterium
Abundance of DS(%)
0
0.01200
0
Abundance of FS(%)
0.01977
0.01977
0
Ochrobactrum
Burkholderia
0.00659
0.00659
Clostridium
0
0
0.04802
Corynebacterium
0.01801
0.00659
Enterobacter
0
0.00659
Enterococcus
0.01801
0.21751
Haemophilus
0
0.00659
Klebsiella
0
0.09228
Legionella
0
0
Moraxella
0
0.01318
Mycobacterium
0.012
0.01318
Pseudomonas
0.006
0
Serratia
0
0.01318
Staphylococcus
0.006
0.00659
Stenotrophomonas
0.006
0.01977
Streptococcus
0.006
0.17796
0.5932