Chemosphere 92 (2013) 1396–

1401

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Chemosphere
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Technical Note

Elimination of high concentration hydrogen sulfide and biogas

purification by chemical–biological process
Kuo-Ling Ho a,1,2
, Wei-Chih Lin a,1
, Ying-Chien Chung b, Yu-Pei Chen b, Ching-Ping Tseng a,⇑
a
Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 300, Taiwan
b
Department of Biological Science and Technology, China University of Science and Technology, Taipei 115, Taiwan

h i G h l i G h t s

● We constructed the chemical–biological H2S removal system in lab and pilot scale.
● The pilot system had operated consecutive 311 d for livestock biogas purification.
● Stable cell density and iron ratio contributed to high H2S removal performance.
● The change of microbial populations was analyzed in pilot scale study.
● This process is feasible for high concentration H2S elimination from biogas.

A R t i c l E I N f O
A B s t R ACt
Article history:
A chemical–biological process was performed to remove a high concentration of H 2S in biogas.
Received 26 November 2012
Received in revised form 3 May
The high iron concentration tolerance (20 g L—1) of Acidithiobacillus ferrooxidans CP9 provided
2013 Accepted 5 May 2013
sufficient ferric iron level for stable and efficient H2S elimination. A laboratory-scale apparatus
Available online 19 June 2013
was setup for a 45 d operation to analyze the optimal conditions. The results reveal that the
H2S removal efficiency reached 98% for
1500 ppm H2S. The optimal ferric iron concentration was kept between 9 and 11 g L—1 with a
Keywords: cell density of 108 CFU g—1 granular activated carbon and a loading of 15 g S m—3 h—1. In pilot-
Bioga scale studies for biogas purification, the average inlet H2S concentration was 1645 ppm with a
s H 2S removal efficiency of up to 97% for a 311 d operation and an inlet loading 40.8 g S m—3 h—1.
Pilot scale When 0.1% glucose was added, the cell density increased twofold under the loading of 65.1
Acidithiobacillus ferrooxidans g S m—3 h—1 with an H2S removal efficiency still above 96%. The analysis results of the
DGGE distribution of microorganisms in the biological reactor by DGGE show
that microorganism populations of 96.7% and 62.7% were identical to the original strain at
day 200 and day 311, respectively. These results clearly demonstrate that ferric iron reduction
by H2S and ferrous iron oxidation by A. ferrooxidans CP9 are feasible processes for the removal of
H2S from biogas.
© 2013 Elsevier Ltd. All rights reserved.

1. Introduction
and efficient, their high cost and production of secondary
pollu- tants are unfavorable.
Corrosive H2S associated with biogas is produced via the
Biological processes that directly metabolize H 2S into
anaer- obic digestion of biodegradable materials, which
sulfate in an efficient and inexpensive way have been
hinders the utili- zation of biogas. Several gas purification
reported. However, the drop in pH caused by sulfate
processes have been applied to eliminate H 2S, such as
accumulation has negative effects (Lee et al., 2006;
chemical scrubbing, physical adsorption (Belmabkhout et
González-Sánchez et al., 2008; Jiang et al., 2009).
al., 2009), electrochemical treatment (Chang and Tseng,
Therefore, studies on biological processes have focused on
1996), and biofiltration (Duan et al., 2007; Ho et al., 2008).
removing low H2S concentrations (10–50 ppm) to prevent
Although physical and chemical treatments are rapid
rapid pH drop (Gabriel, 2003; Kim et al., 2008; Goncalves and
Govind, 2009; Ryu et al., 2009; Ramírez et al., 2011).
Other studies that combined chemical and biological
* Corresponding author. Address: College of Biological Science and processes for both H2S elimination and ferric iron
Technology, National Chiao Tung University, 75 Bo-Ai Street, Hsin-Chu,
regeneration by Acidithiobacillus ferrooxidans have been
Taiwan. Tel.: +886 3 5731596; fax: +886 3 5729288.
E-mail address: [email protected] (C.-P. Tseng). reported (Giro et al., 2006; Alemzadeh et al., 2009). These
1
These authors equally contributed to this paper. processes are based on two reactions as follows: the inlet
2
Current address: Department of Chemical Engineering, National Taiwan H2S is first oxidized with a ferric iron solution and yields
Univer- sity, Taipei 106, Taiwan. elemental sulfur, and the reduced ferrous iron is then
reoxidized by A. ferrooxidans in the biological process.

0045-6535/$ - see front matter © 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.chemosphere.2013.05.054
 K.-L. Ho et al. / Chemosphere 92 (2013) 1396–1401 1397
height is 150 cm. The biogas produced from the anaerobic
Separate studies that focused on chemical absorption (Asai wastewater treatment system was pumped into the
et al., 1990; Ebrahimi et al., 2003) and biological oxidation chemical reactor at specific flow
(Mousavi et al., 2008; Alemzadeh et al., 2009) were
proposed. For instance, when the inlet H2S concentration is
lower than 300 ppm, the rate-limiting step for the chemical
absorption process is the mass-transfer limitation, as
revealed by model validation (Chung et al., 2003). When
the system has a high inlet H2S concentration (e.g., above
1500 ppm), the H2S removal efficiency is significantly
maintained via long GRT (gas retention time) and stable ferric
iron concentration in the chemical reactor (Chung et al.,
2006). In the biological reaction, Mesa et al. (2004)
investigated the continuous oxidation efficiency of ferrous
iron by using immobilized A. ferroox- idans. However, studies
that focused on the combined and contin- uous operation
are limited.
In the present study, the chemical absorption reactor
and the biological oxidation reactor with immobilized A.
ferrooxidans CP9 were connected and then examined in both
laboratory and pilot scales to evaluate the performance of
H2S elimination. In the laboratory-scale study, optimal
operating parameters such as GRT, temperature, and H 2S
inlet loading were examined. In the pilot-scale study, the
biogas with an average H 2S concentration of 1645 ppm
was introduced into the chemical–biological system. The
long-term performance was examined, and the results
demon- strate that the chemical–biological process
effectively removed H2S from the biogas.

2. Material and methods

2.1. Microorganism and cultivation

A. ferrooxidans CP9 was isolated from an acid mine

drainage, and then identified by the 16S rRNA gene
(GenBank accession number EF605251). This strain was
grown in the M16 broth med- ium (Kim et al., 2002). A solid
KBU medium was prepared for cell number counting of A.
ferrooxidans CP9 (Khalid et al., 1993).
In the immobilization process, granular activated carbon
(GAC) with a diameter of 4.5 mm and a surface area of
1250 m2 g—1 was used (China Activated Carbon Industries,
Taiwan) to immobilize the A. ferrooxidans CP9 in the
bioreactor. The details of the proce- dure were described
in a previous report (Chung et al., 2006).
The cell-laden GAC was transferred to the bioreactor when
the cell density reached 108 CFU g—1 GAC.

2.2. Combined chemical–biological reactor for H2S elimination

A scheme of the combined chemical–biological system

on a lab- oratory scale is shown in Fig. 1. The 1% H2S gas,
which was supplied from a gas cylinder, was initially diluted
with compressed air, and then pumped into the bottom of
the chemical reactor. The glass chemical reactor (12 cm
id × 50 cm H) with a height of 40 cm
was filled with the M16 medium, which contained glass
beads
(id: 6 mm). A perforated sieve plate at the bottom of the
reactor al- lowed the H2S gas to flow in. The bioreactor (12
cm id × 50 cm H) with a height of 40 cm was packed
with GAC to immobilize
A. ferrooxidans CP9 in the M16 medium. The bioreactor was
fitted with a thermostatic jacket to control the temperature. A
funnel- shaped sulfur separation tank was set between the
chemical and biological reactors to collect the elemental
sulfur. The circulation of the medium between each column
was driven by a peristaltic pump at specific circulating rates.
The composition of the pilot scale chemical–biological
reactor was similar to that shown in Fig. 1, except the
rates. The maintenance of liquid level and glucose
concentration in this system was checked weekly during the
operation.

2.3. DGGE and phylogenetic analysis

The PCR primers of 968f GC and 1401r were used to

amplify the segment of the eubacterial 16S rDNA (Brosius
et al., 1981). The process of manipulating the extracted
genomic DNA and the amplified PCR is shown in a
previous study (Ho et al., 2008). The samples were run on
8% acrylamide gel with a denaturant gradient between 45%
and 60% by using a Bio-Rad DGGE apparatus. The
electrophoresis procedure was run, stained, and rinsed,
as de- scribed in a previous report (Torsvik et al., 2000).
The interesting fragments were excised from the DGGE gels
for DNA amplification and sequencing. Phylogenetic trees
were constructed from the evolutionary distance by using
the Tree View software.

2.4. Analytical methods

H2S concentrations were measured using gas detector

tubes (Kitagawa). Ferrous, ferric, and total iron
concentrations were determined using a previously
described method (Chung et al., 2006). For cell density
analyses, 0.5 g of GAC was collected from the bioreactor
and vortexed with 5 mL of sterile water for 5 min, and
sprayed on the solid KBU medium. The dark-brown colony
in the medium was indicated as A. ferrooxidans CP9 after 7–
14 d of cultivation.

3. Results and discussion

3.1. Effects of inlet H2S concentration on removal efficiency and Fe2+/

Fe3+ ratio in the laboratory scale

The influence of H2S concentration on chemical absorption

behavior was examined in the laboratory-scale combined
reactor. Fig. 2a shows the effect of inlet H2S concentration on
the efficiency of the system during the 45 d operation with a
GRT of 8 min. The results indicate that the removal efficiency
increased with decreas- ing inlet H 2S concentration during
the first 15 d. A removal effi- ciency of 98% was achieved
when the inlet H2S concentration was less than 1500 ppm.
Fig. 2b and c show the variation in Fe 2+/ Fe3+ ratio and pH
during the 45 d operation. The ferric iron con- sumption rate
and the ferrous iron accumulation rate were propor- tional to
the inlet H2S concentration. In the high loading stage

Fig. 1. A schematic of the laboratory scale combined chemical–biological

reactor for H2S treatment: (1) biological column, (2) chemical column, (3)
liquid peristaltic pump, (4) sulfur storage tank, (5) air pump, (6) gas flow
meter, (7) H2S gas cylinder,
(8) inlet of H2S, and (9) inlet of air.
1398 K.-L. Ho et al. / Chemosphere 92 (2013) 1396–1401

(2000 ppm), the H2S removal efficiency decreased when

removal efficiency. The ferric iron regeneration rate in the
the ferric iron decreased to 6 g L—1 in the chemical reactor.
On the other hand, when H2S was fed at 1000 ppm, the biolog- ical reactor was insufficient to support H 2S oxidization
ferrous and ferric iron concentrations remained stable in the chemical reactor during day 65–90 (Fig. 3c). In the first
between 9 and 11 g L—1 (Fe2+/ Fe3+ = 1 ± 0.2) in both 30 d, the H2S removal efficiency reached 100% when the
reactors and then reached a removal efficiency of over ferric iron concen-
99%. Therefore, when GRT was kept at 8 min, the system tration was about 10 g L—1, but continuously dropped to
maintained the Fe2+/Fe3+ ratio and achieved high removal less than 90% when the ferric iron concentration was less
than 6 g L—1. This result is consistent with those of Mesa et
efficiency when the inlet H2S concentration was between al. (2004), who concluded
1000 and 1500 ppm. Giro et al. (2006) treated a gas with a
that the maximum H2S absorption efficiency is reached
high H2S when the ferric iron concentration is between 10 and 13 g
concentration (20000 ppm) by using a combined chemical– L—1, but remark- ably decreased in the lower concentration
biological system; however, they used a lower H2S region.
loading (10.9 g S m—3 h—1) and a shorter operation time In this laboratory-scale combined system, an H2S
(600 min) com- pared with this study. loading over 40 g S m—3 h—1 decreased the H2S removal
The effect of inlet loading change on biological behavior efficiency to below 90% because of ferric iron consumption.
was also examined. The variations in pH (pH 1.7–1.9) and A previous report (Pagella and De Faveri, 2000) applied a
cell density (6 × 107 to 7 × 107 CFU g—1 GAC) were stable loading of 66 g S m—3 h—1 in a com- bined system.
despite the H2S inlet concentrations. After the shock However, the inlet H2S concentration was low
loading test (from day 10 to 15), (100 ppm), and the removal efficiency dropped to 30%
the system showed a remarkable ability for recovery. after the 12 d operation due to the ferric iron concentration
Although the ferrous iron concentration was continuously dropped. This result was attributed to the
continuously kept above 14 g L—1 during the shock loading insufficient biological oxidative ability of the bioreactor and
test, cell growth was not nega- tively affected.
the failure to remove sulfur precipitates from the chemical
absorber; hence, both factors re- duced the mass transfer
3.2. Effects of GRT on H2S removal efficiency and Fe2+/Fe3+ efficiency of the liquid–H2S interface (Pagella and De
concentration in the laboratory scale Faveri, 2000).

In the 90 d continuous experiment, the GRT was

gradually decreased to increase the H2S loading value. Fig. 3
shows the effect of GRT on the H2S removal efficiency and
Fe2+/Fe3+ concentration at an inlet H2S concentration of 1000
ppm. The results indicate that the H2S removal efficiency
dropped to 82% when the GRT was 4 min (Fig. 3a), and
further dropped to 15% when the GRT was 1 min. The
ferric iron concentration in the chemical reactor decreased
with decreasing GRT (Fig. 3b) and caused a drop in the

Fig. 2. Laboratory scale H2S elimination in various inlet concentrations of

the 45 d operation. (a) H2S inlet/outlet concentration and removal Fig. 3. Laboratory scale H 2S elimination in various GRT of the 90 d
efficiency. The iron concentration plot in (b) chemical reactor and (c) operation. (a) Loading and removal efficiency plot. The iron concentration
bioreactor. profile in (b) chemical reactor and (c) bioreactor.
 K.-L. Ho et al. / Chemosphere 92 (2013) 1396–1401 1399

3.3. Effect of temperature on system performance in the laboratory

scale

Gomez et al. (2000) found that the ferrous iron

oxidation effi- ciency of A. ferrooxidans dramatically
decreases when the temper- ature was increased from 30
to 40 °C. In addition, the ferrous iron oxidation efficiency of
A. ferrooxidans was weak at 40 °C in the bio- reactor (Chung
et al., 2003). We conducted a 30 d test to analyze further
the effects of temperature on the performance of the com-
bined system. The six stages were 35, 26, 18, 26, 42, and 26
°C, and
each stage lasted for five days. The experimental
conditions were kept at 8 min GRT, 1,500 ppm H2S
concentration (15 g S m—3 h—1 loading), and 3 L min—1
medium circulation rate. The results show that the removal
efficiency reached 99% in all stages except at
42 °C, which was slightly lower at 97%. The high efficiency
was likely due to the buffering ability of chemical adsorption.
Although the ferric iron concentration declined at 42 °C, it
remained greater than 8 g L—1 and was enough to oxidize
H2S efficiently. Neverthe- less, the number of A. ferrooxidans
CP9 cells decreased to 1/2 and
1/8 at 18 and 42 °C, respectively, compared with that at
35 °C. Apparently, the relatively low cell density in the
bioreactor at 42 °C slowed the ferrous iron oxidation
efficiency and slightly de- creased the H2S removal
efficiency. The oxidation efficiencies of 20.1, 33.4, 29.2,
and 12.6 mg L—1 h—1 were determined at condi-
tions of 18, 26, 35, and 42 °C, respectively. The different oxidation
efficiencies explain the minor variations in H 2S removal
efficiency. Moreover, the results show that the combined
chemical–biological system could be applied within a broad
temperature range.

3.4. Effect of loading and carbon source on biogas purification in the

pilot-scale study

The biogas composition (%) in this study was 61 ± 9 CH4, consumption rate in the
26 ± 4 CO2,4 ± 1 O2, and 1645 ± 345 ppm H 2S in average chemical process. However, the cell density increased to 9 ×
during the 311 d continuous operation. The GRT was kept at 3 108 - CFU g—1 GAC with 0.1% glucose and then neutralized
the high
min in the first 150 d and then reduced to 2 min during day
H2S loading effect. Therefore, the addition of glucose
151–311. The liquid circula- tion rate was maintained at 2
caused the variation in ferric iron concentration in the
mL min—1, and the initial Fe2+/Fe3+
chemical reactor, which
concentrations were both at 10 g L—1. Fig. 4a shows the
H2S re-
moval efficiency at different H 2S concentrations ranging
from 890 to 2250 ppm in the inlet biogas. The average H2S
removal effi- ciencies for the stages without (day 1–90) and
with (day 91–311) glucose were both about 98%.
The GRT decreased to 2 min during day 151–311,
although the increased H2S loading led to the accumulation
of ferrous iron, the ferric iron concentration remained
above 6 g L—1, and the average H2S removal efficiency was
sustained at 97%. In previous studies, biofilters with
shorter GRTs (1.6–30 s) were applied in the field
to treat waste gases containing H2S (Gabriel, 2003; Duan
et al., 2005, 2007). However, the inlet H2S concentrations
were operated below 100 ppm in these studies.
In this pilot-scale operation, the ferric iron concentration in
the chemical reactor was initially kept stable in the first 50 d
and then slightly decreased from 10 to 8.4 g L—1 during day
50–90. However,
it recovered to 9.2 g L—1 in the next 60 d after the addition
of 0.1%
glucose (Fig. 4b and c). The average cell density with
glucose was 8 × 108 CFU g—1 GAC, which was twofold higher
than that without glucose. Therefore, the elevated ferric iron
concentration after the addition of 0.1% glucose (day 50–90)
was attributed to the increase in cell density in this
operation.
During day 151–311 of high H2S loading (65.1 g S m—3 h
—1
on the average), the formation rate of ferric iron in the
bioreactor was not fast enough compared with the
Fig. 4. Biogas purification in the pilot scale of 311 d operation. (a) H2S
inlet/outlet concentration and removal efficiency. (b) The iron
concentration plot in chemical reactor. (c) The iron concentration and cell
density plot in bioreactor.

merely decreased from 9.2 to 7.7 g L—1. This result was in

agree- ment with a previous laboratory-scale study,
where the addition of glucose improved the iron
oxidation ability of A. ferrooxidans
(Chung et al., 2006).

3.5. Analysis of the microorganism community during pilot-scale

operation

PCR-DGGE was used to determine the variations in

bacterial communities. Fig. 5 shows the profile of the DGGE
bands during the 311 d operation. The complexity of the
DGGE bands increased with increasing operating time. These
bands show that exotic bac- terial populations appeared
after glucose was added. However, band B (inoculated A.
ferrooxidans CP9) was a consistently domi- nant population in
the bacterial consortium throughout the opera- tion time.
The intensity of band B on day 90 was 100%, which indicates
that only A. ferrooxidans was detected in this system. Three
minor microbes that comprise bands C (0.9%), J (0.2%), and
K (2.2%) appeared on day 200, but the intensity of band B
remained as high as 96.7%.
On the day 311, ten discernible bands were observed,
namely, bands A (4.3%), B (62.7%), C (5.4%), D (1.7%), E
(7.3%), F (2.4%), G
(6.8%), H (6.7%), I (1.7%), and J (1%). Band B was the most
intense and predominant band, which was identified as A.
ferrooxidans (99%). Bands C and H belonged to phylum
proteobacteria. Their close relatives shared close
similarities to Acidithiobacillus sp.
1400 K.-L. Ho et al. / Chemosphere 92 (2013) 1396–1401

Fig. 5. Analysis of the microorganism community during pilot scale operation. (a) DGGE analysis for day 90, 200, and 311. (b) Plot of the phylogenetic
trees.

ZJJN-3 (98%) and Halothiobacillus neapolitanus strain CIP 104769 increased the cell density of
(99%). Three bands (E, G, and I) were clustered to the phylum
Acidobacteria, namely, Terriglobus sp. TAA 43 (97%), uncultured
bacterium mle1-25 (98%), and uncultured bacterium clone
BS69 (96%). Two bands (A and D) were grouped to the
phylum Firmicutes, namely, Sulfobacillus acidophilus strain
GG6/1 (99%) and Alicyclobacillus acidocaldarius strains 1–4 (98%).
Bands F and J were too weak and could not be identified.
According to these results, all sequences were identified as
aer- obic and acidophilic bacteria with an optimal growth
pH ranging from 2 to 4. Most microbes were autotrophic,
except for S. acidoph- ilus (band A), A. acidocaldarius (band D),
and Terriglobus sp. (band E), which were mixotrophic,
heterotrophic, and oligotrophic, respec- tively (Eichorst et
al., 2007). This result suggests that the addition of glucose
increased the complexity of the microbial communities.
Johnson et al. (2005) showed that S. acidophilus (band A), which
was similar to the inoculated A. ferrooxidans, possesses
reducing power from oxidizing ferrous iron or sulfur.
Besides, the H. neapolitanus (band H) was designated as
a sulfur-oxidizing bacterium (García-de-la-Fuente et al.,
2011). Minor elemental sulfur formed by the H2S oxidation
reaction in the chemical reactor was expected to migrate to
the bioreactor through medium circulation and could be an
energy source to some exclusively sulfide-oxidizing
bacteria. Although the extra carbon source gave rise to the
appearance of exotic microbes in the system, it also
the inoculated strain by twofold compared with the cell
density in the earlier 90 d of the operation. On day 311, A.
ferrooxidans was still dominant at 62.7%, and the H 2S
removal efficiency was maintained at 96%. This result
showed that the ferrous oxidation ability of this system did
not decrease with increasing microbial complexity.

4. Conclusions

We demonstrated that the chemical–biological

process immo- bilized with iron-tolerant A. ferrooxidans
CP9 maintained the balance of the Fe2+/Fe3+ ratio and
reached an H2S removal efficiency of 98%. In the pilot-
scale operations, the addition of glu- cose improved the
biogas purification efficiency by increasing the cell
density and ferrous oxidation efficiency. The H 2S loading
reached 65.1 g S m—3 h—1 (3.3-fold higher than the
laboratory-scale
condition) with a removal efficiency of 96%. In addition, the
factors
of high tolerance for iron ions at 20 g L—1 and the rapid
ferrous iron oxidation ability of A. ferrooxidans CP9 were
important in main-
taining the balance of Fe2+/Fe3+ concentrations. Although
the exotic microbes appeared during the 311 d
operation, the A. ferrooxidans CP9 cell density remained
more than 108 CFU g—1 GAC and offered a stable ferrous
iron oxidation ability. These results clearly demon-
strate that the chemical–biological process is a feasible
method for eliminating high H2S concentration from
biogas.
 K.-L. Ho et al. / Chemosphere 92 (2013) 1396–1401 1401

Acknowledgements Giro, M.E.A., Garcia Jr., O., Zaiat, M., 2006. Immobilized cells of Acidithiobacillus
ferrooxidans in PVC strands and sulfite removal in a pilot-scale bioreactor.
Chem. Eng. J. 28, 201–207.
This work was financially supported by grants NSC 100-
Gomez, J., Cantero, D., Webb, C., 2000. Immobilisation of Thiobacillus
3114-B- 009-001 from the National Science Council and by ferrooxidans cells on nickel alloy fibre for ferrous sulfate oxidation. Appl.
the Aim for the Top University Plan of the National Chiao Tung Microbiol. Biotechnol. 54, 335–340.
University and Ministry of Education, Taiwan. Goncalves, J.J., Govind, R., 2009. Enhanced biofiltration using cell attachment
promotors. Environ. Sci. Technol. 43, 1049–1054.
González-Sánchez, A., Revah, S., Deshusses, M.A., 2008. Alkaline biofiltration
References of H2S odors. Environ. Sci. Technol. 42, 7398–7404.
Ho, K.L., Chung, Y.C., Lin, Y.H., Tseng, C.P., 2008. Microbial populations
Alemzadeh, I., Kahrizi, E., Vossoughi, M., 2009. Bio-oxidation of ferrous ions by analysis and field application of biofilter for the removal of volatile-sulfur
Acidithioobacillus ferrooxidans in a monolithic bioreactor. J. Chem. Technol. compounds from swine wastewater treatment system. J. Hazard. Mater.
Biotechnol. 84, 504–510. 152, 580–588.
Asai, S., Konishi, Y., Yabu, T., 1990. Kinetics of absorption of hydrogen sulfide Jiang, X., Yan, R., Tay, J.H., 2009. Simultaneous autotrophic biodegradation of
into aqueous ferric sulfate solutions. AIChE J. 36, 1331–1338. H2S and NH3 in a biotrickling filter. Chemosphere 75, 1350–1355.
Belmabkhout, Y., De Weireld, G., Sayari, A., 2009. Amine-bearing mesoporous Johnson, D.B., Okibe, N., Hallberg, K.B., 2005. Differentiation and identification
silica for CO2 and H2S removal from natural gas and biogas. Langmuir 25, of iron-oxidizing acidophilic bacteria using cultivation techniques and
13275– 13278. amplified ribosomal DNA restriction enzyme analysis. J. Microbiol. Methods 60,
Brosius, J., Dull, T.J., Sleeter, D.D., Noller, H.F., 1981. Gene organization and 299–313. Khalid, A., Bhatti, T., Umar, M., 1993. An improved solid
primary structure of a ribosomal RNA operon from Escherichia coli. J. Mol. medium for isolation, enumeration and genetic investigations of
Biol. 148, 107– 127. autotrophic iron-and sulphur-
Chang, M., Tseng, T., 1996. Gas-phase removal of H 2S and NH3 with dielectric oxidizing bacteria. Appl. Microbiol. Biotechnol. 39, 259–263.
barrier discharges. J. Environ. Eng. 122, 41–46. Kim, J.H., Rene, E.R., Park, H.S., 2008. Biological oxidation of hydrogen sulfide
Chung, Y.C., Ho, K.L., Tseng, C.P., 2003. Hydrogen sulfide gas treatment by a under steady and transient state conditions in an immobilized cell biofilter.
chemical–biological process: chemical absorption and biological Bioresour. Technol. 99, 583–588.
oxidation steps. J. Environ. Sci. Health, Part B 38, 663–679. Kim, T.W., Kim, C.J., Chang, Y.K., Ryu, H.W., Cho, K.S., 2002. Development of
Chung, Y.C., Ho, K.L., Tseng, C.P., 2006. Treatment of high H 2S concentrations an optimal medium for continuous ferrous iron oxidation by immobilized
by chemical absorption and biological oxidation process. Environ. Eng. Sci. Acidothiobacillus ferrooxidans cells. Biotechnol. Prog. 18, 752–759.
23, 942–953. Lee, E.Y., Lee, N.Y., Cho, K.-S., Ryu, H.W., 2006. Removal of hydrogen
Duan, H., Koe, L.C.C., Yan, R., 2005. Treatment of H2S using a horizontal sulfide by sulfate-resistant Acidithiobacillus thiooxidans AZ11. J. Biosci.
biotrickling filter based on biological activated carbon: reactor setup and Bioeng. 101, 309– 314.
performance evaluation. Appl. Microbiol. Biotechnol. 67, 143–149. Mesa, M., Andrades, J., Macias, M., Cantero, D., 2004. Biological oxidation of
Duan, H., Yan, R., Koe, L.C.C., Wang, X., 2007. Combined effect of adsorption ferrous iron: study of bioreactor efficiency. J. Chem. Technol. Biotechnol.
and biodegradation of biological activated carbon on H 2S biotrickling 79, 163–170. Mousavi, S.M., Yaghmaei, S., Vossoughi, M., Roostaazad, R.,
filtration. Chemosphere 66, 1684–1691. Jafari, A., Ebrahimi, M., Chabok, O.H., Turunen, I., 2008. The effects of Fe(II)
Ebrahimi, S., Kleerebezem, R., van Loosdrecht, M., Heijnen, J., 2003. Kinetics of and Fe(III) concentration and initial pH on microbial leaching of low-grade
the reactive absorption of hydrogen sulfide into aqueous ferric sulfate sphalerite ore in a column
solutions. Chem. Eng. Sci. 58, 417–427. reactor. Bioresour. Technol. 99, 2840–2845.
Eichorst, S.A., Breznak, J.A., Schmidt, T.M., 2007. Isolation and characterization Pagella, C., De Faveri, D., 2000. H2S gas treatment by iron bioprocess. Chem.
of soil bacteria that define Terriglobus gen. nov., in the phylum Acidobacteria. Eng. Sci.
Appl. Environ. Microbiol. 73, 2708–2717. 55, 2185–2194.
Gabriel, D., 2003. Retrofitting existing chemical scrubbers to biotrickling Ramírez, M., Fernández, M., Granada, C., Le Borgne, S., Gómez, J.M., Cantero,
filters for H2S emission control. Proc. Natl. Acad. Sci. 100, 6308–6312. D., 2011. Biofiltration of reduced sulphur compounds and community
García-de-la-Fuente, R., Cuesta, G., Sanchís-Jiménez, E., Botella, S., Abad, M., analysis of sulphur-oxidizing bacteria. Bioresour. Technol. 102, 4047–
Fornes, F., 2011. Bacteria involved in sulfur amendment oxidation and 4053.
acidification processes of alkaline ‘‘alperujo’’ compost. Bioresour. Ryu, H.W., Yoo, S.-K., Choi, J.M., Cho, K.-S., Cha, D.K., 2009. Thermophilic
Technol. 102, 1481–1488. biofiltration of H2S and isolation of a thermophilic and heterotrophic H 2S-
degrading bacterium, Bacillus sp. TSO3. J. Hazard. Mater. 168, 501–506.
Torsvik, V., Øvreås, L., Nakatsu, C.H., 2000. Soil community analysis using
DGGE of 16S rDNA polymerase chain reaction products. Soil Sci. Soc. Am.
J. 64, 1382– 1388.