Fuel 82 (2003) 13591365

www.fuelfirst.com

Empirical model for methane oxidation using a composted

pine bark biofilterq
Chris A. du Plessisa,*, Johannes M. Straussb, Elijah M.T. Sebapalob, Karl-Heinz J. Riedelb
b

a
BHP Billiton, Private Bag X10014, Randburg 2125, South Africa
School of Environmental Sciences and Development: Microbiology, Potchefstroom University for Christian Higher Education,
Private Bag X6001, Potchefstroom 2520, South Africa

Received 27 August 2002; revised 4 February 2003; accepted 5 February 2003; available online 10 March 2003

Abstract
Methane concentration in the explosive range is of particular concern in the confined space of underground mining environments, where
the hazards of accidental ignition of methane may be further compounded by coal dust explosions. The aim of this study was to determine
whether composted pine bark, could be used as biofiltration support media for methane oxidation and to determine the degradation rates of
methane concentrations approaching the explosive range (i.e. 5.5% v/v methane). Although of organic origin, composted pine bark is mainly
composed of non-labile and recalcitrant large molecular weight molecules due to the fact that the labile organic compounds in the bark are
oxidised during the composting process. Methane removal efficiencies in a composted pine bark biofilter were determined at methane
concentrations ranging from 0.1 to 2.5% (v/v) and retention times of 20 400 min. Methane removal efficiencies exceeding 70% were
obtained when the biofilter was subjected to gas retention times in excess of 30 min and methane concentrations up to 0.5% (v/v). The data
obtained were used to develop an empirical model that successfully described the overall removal efficiency with an R2 value of 0.97. It was
concluded that composted pine bark could indeed be successfully utilised in passive gas-flow biofiltration for the attenuation of methane in
worked underground coal mining chambers.
q 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Biofiltration; Composted pine bark; Empirical model; Methane

1. Introduction
Methane (CH4), as the simplest hydrocarbon, is often
encountered in both natural and man-made environments.
It is the major constituent of natural gas and is also present
in many coal formations. Although stable in anaerobic
environments, methane poses a distinct explosion hazard
when in the range 5.53 14% (v/v) in the presence of oxygen
[1,2]. Methane concentration, in the explosive range, is of
particular concern in the confined space of underground
mining environments where the hazards of accidental
ignition of methane may be further compounded by coal
dust explosions [3]. Current practice in such coal mining
environments is to utilise methods of degasification ahead of
the mining face operation together with ventilation to
dilute the remaining methane in worked underground
* Corresponding author. Tel.: 27-11792-7090; fax: 27-11792-7097.
E-mail address: chris.duplessis@bhpbilliton.com (C.A. du Plessis).
q
Published first on the web via Fuelfirst.comhttp://www.fuelfirst.com

chambers [2,4]. The requirement for continued long-term

ventilation of underground chambers poses a considerable
operating cost for, particularly, deeper coal mining
operations.
An alternative approach to methane ventilation in
worked underground mine chambers, is the bacterial
oxidation of methane to carbon dioxide. Such biofiltration
treatment systems are commonly applied to a wide range
of volatile compounds where destruction of targeted
compounds are achieved by passing through a porous
support media onto which pollutant-degrading microorganisms are immobilised [5]. Despite the vast body of literature
detailing the oxidation of methane in natural environments
[6 8] relatively little research has been conducted
into the application of methane biofiltration [3,4].
Methane oxidation in biofiltration systems is less suitable
than for many other volatile organic compounds (VOC)
mainly due to its relative recalcitrance, low level of water
solubility [9] and the fact that the bacterial strains capable of

0016-2361/03/$ - see front matter q 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0016-2361(03)00040-1

1360

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

its oxidation are less abundant that those capable of

degrading other VOCs such as toluene [8,10].
The aim of this study was to determine whether
composted pine bark could be used as biofiltration support
media for methane oxidation and to determine the
degradation rates of methane concentrations approaching
the explosive range. The known relative recalcitrance of
methane to microbial oxidation, would mean that any
microbial treatment process would be more effective
at lower concentrations. The methane concentrations
in underground mined chamber atmospheres typically
gradually increase with time. Based on potential future
implementation of methane biofiltration treatment systems,
the aim of this investigation was to treat methane at
concentrations in the range 0.1 2.5%, thereby preventing
increased methane concentration that would approach the
explosive range. Because of the known recalcitrance of
methane the potential impact of a methane biofiltration
system was believed to be greater in the 0.1 2.5% rather
than in the 2.5 5.5% range. Composted pine bark was
specifically selected as a potential biofilter support media
for several reasons. The composting process typically takes
4 8 months during which aeration, moisture content and
nutrient additions are optimally maintained for microbial
growth. The presence of diverse organic compounds
contained in pine bark [11,12] results in the proliferation
of bacteria and fungi capable of oxidising a large number
of these diverse compounds and is, therefore, often
successfully utilised as biofiltration support media [13].
From available literature, it was evident that biofiltration of
methane concentrations approaching the lower explosive
limit, would not be achieved within the relatively short
reactor residence times (, 1 min) possible when treating
other carbon emission [14]. The requirement for lengthy
reactor residence times, and the vast reactor size requirements that would apply to underground mining environments, necessitated the use of a relatively inexpensive and
readily available biofiltration support medium. In addition,
the biofiltration support media would need to be composed
mainly of non-labile carbon compounds in order to ensure
long-term durability and integrity. Composted pine bark
was selected based on these criteria and was specifically
not inoculated with bacterial species known for methane
oxidation as this would be impractical on a larger scale and
in order to determine whether the natural population
contained in the composted pine bark would exhibit
methane oxidation. The use of this media was also
considered based on reports that natural environments
may contain bacterial strains with lower half-saturation
values for methane oxidation than pure colonies cultured
under methane-rich conditions [15]. During the composting process the labile components in the pine bark are
oxidised and polymerised, rendering a composted pine
bark fraction composed mainly of lignin-type molecules
that are not readily further decomposed [16,17], thus,
making the composted pine bark a suitable biofilter support

medium. This study was also aimed at the development of

an empirical model for the biodegradation of methane
suitable for practical implementation in coal mining
environments.

2. Materials and methods

2.1. Reactor description
The experimental reactor used during this study
consisted of five PVC sections (Fig. 1), each with an
inner diameter of 17 cm and a height of 30 cm. Four of the
five sections were packed with support media, with the
lower section serving as a liquid reservoir. Each section,
separated by a stainless steel sieve located at the bottom,
was packed to 5 cm from the top, forming a plenum to
allow gas flow redistribution. Gas sampling points were
located in each of the plenums, as well as at each of the
inlet and outlet ports of the reactor column. Two separate
air streams entered a mixing chamber with similar
dimensions to one of the reactor sections. One air stream,
set to a specific flow rate and connected to a three-way
solenoid valve and timer, periodically supplied a nebuliser
(Hospitak Upmist, Lindenhurt, NY) with ambient air to
produce a mineral salt medium aerosol. The mineral salt
medium also served to maintain a neutral pH in the reactor,
as this pH has been shown to be most conducive to
methane oxidation [10]. The other air stream containing
the methane gas (Afrox, South Africa) was premixed
outside the reactor to obtain specific methane gas
concentrations and retention times inside the reactor.
This air stream was humidified by bubbling through
water. Both air streams converged in the mixing chamber
prior to entering the reactor. The reactor was operated in a
directional-switching mode, switched every 3.5 days, to
overcome non-uniform bacterial distribution on the support
media [14].
2.2. Mineral salt medium composition
The mineral salt medium used in the study had the
following composition: Na 2 HPO 4, 4 g l 21; K2 HPO 4,
1.5 g l21; NH4Cl, 1 g l21; MgSO47H2O, 0.2 g l21; CaCl2,
0.01 g l21 and FeNH4-citrate, 0.005 g l21.
2.3. Biofilter support media
The reactor was packed with composted pine bark
(fraction sizes 5 12 mm) (Culterra (Pty) Ltd, South
Africa) mixed with one-third (volume-to-volume) Perlite
(Genulite) (fraction sizes 3 5 mm) (Chemserve Perlite,
Parklands, South Africa). This resulted in a packing
volume of 20.26 l and air fill porosity of 13.12 l (35.2%
air fill porosity).

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

1361

Fig. 1. Schematic representation of the methane biofilter set-up.

2.4. Microbial cultivation and identification

The reactor was acclimatised to methane oxidation over a
period of 3 months during which it was continuously
exposed to an air stream supplemented with 30% methane at
a gas retention time of 100 min. After this acclimation
period the media was sampled for culturable methane
oxidising bacteria by suspending in sterile saline solution
(0.85% NaCl) to extract a portion of the attached biofilm
bacteria. The solution was streaked onto mineral salt
medium agar plates and incubated in a 30% methane
atmosphere at 25 8C. Colonies were purified by continuous
sub-culturing under similar conditions. Pure colonies were
identified using PCR amplification, cloning and 16S rDNA
sequencing techniques [18].
2.5. Analytical methods
Gas samples were collected from the biofilter in 5 ml
gas-tight sampling syringes (Hamilton Series #1005)
equipped with Teflon Mininert w fittings. Samples
were analysed using a HP 6850 gas chromatograph
(Hewlett-Packard Co.) equipped with a 1 ml heated
(160 8C) gas sample loop and a flame ionisation detector

(FID). A HP-5 (Hewlett-Packard Co.) column

(30 m 0.25 mm) was used with N2 as the carrier gas
at a flow rate of 1 ml min21. The column temperature
was maintained at 70 8C, the injector at 160 8C and the
detector at 280 8C. All samples were injected in split
mode with a split ratio of 5:1.
2.6. Methane concentrations, retention times
and removal efficiencies
The effects of methane concentrations, in the range
0.1 2.5% (v/v), on biofilter removal efficiencies were
evaluated at gas retention times ranging from 20 to
400 min. Actual gas retention times were calculated from
the bed porosity of the support medium multiplied by the
total bed volume divided by the gas flow rate.
This parameter, rather than the frequently used empty
bed residence time (EBRT), was used in this study.
The outlet removal efficiencies were determined at steady
state conditions by comparing inlet with outlet concentrations. Steady state, or pseudo-steady state, conditions
were assumed when such conditions were reflected by
consistent methane oxidation efficiencies over a period of
several days, at each test condition.

1362

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

Parameters a and b could be related to prevailing methane

concentrations (% v/v) using Eqs. (2) and (3) for
parameters a and b:

3. Results and discussion

3.1. Methane oxidizing bacteria
The culturable methane oxidising strains in the
reactor were identified as Flexibacter scanti, Pseudomonas fluorescens and Pseudomonas aeruginosa. F. scanti,
a member of the Cytophaga Flexibacter Bacteroides
phylum, has previously been shown to be ubiquitously
associated with methane-containing environments [19,20]
while members of the Pseudomonas genus have also
frequently been associated with methylotrophic activity
[21,22]. The fact that well-known methane oxidizing
bacteria were not detected could be ascribed to the
non-selective growth environment provided for by
the presence of composted pine bark. This medium
contributes numerous bark-constituent carbon sources,
thus resulting in the proliferation of bacteria not
exclusively dependent upon methane oxidation as a
carbon source.
The sub-culturing purification technique in a methane
atmosphere used in this study did introduce an element of
selectivity, thereby limiting the identified species to those
that are culturable in pure colonies. Although direct
molecular biology identification techniques may have
overcome the bias introduced by the culturing step,
such techniques suffer from the inability to determine the
identity of bacteria specifically responsible for methane
oxidation. While the culturing step used in this study did
not facilitate comprehensive elucidation of the methane
oxidizer population dynamics, it did serve to confirm the
ubiquitous presence of some methane oxidizing bacteria in
composted pine bark. This finding confirms the suitability of
composted pine bark as a biofiltration support medium for
use in large-scale reactors in underground mine
chambers without the need for specific inoculation with
methanotrophic bacteria. The microbial, as apposed to
abiotic, elimination of methane within the biofilter was
confirmed by the replacement of air with nitrogen in the
reactor feed gas mixture, and observing the immediate
elimination of methane oxidation.
3.2. Model development
The methane removal efficiencies (RE) for the outlet
(bed height of 1.0 m) were determined at five different
methane concentrations C and retention times t ranging
from 20 to 400 min. The removal efficiencies as a
function of retention time, at various methane concentrations, are illustrated in Fig. 2(a e). The curve fitting
parameters a and b (Eq. (1)) used to describe the
relationships between the methane removal efficiencies
and retention times, at different methane concentrations,
are given in Table 1.
RE a1 2 e2bt

a 26:3637 lnC 67:568

b 20:0157 lnC 0:0338

Substitution of Eqs. (2) and (3) into Eq. (1), provided an

empirical model (Eq. (4)) that described the removal
efficiency of methane as a function of gas retention time
and methane concentration.
RE 26:3637 lnC 67:568
1 2 e2t20:0157 lnC0:0338

This empirical model had an overall R2 of 0.97 when

fitted to the entire data set. The reactor performance as a
function of methane concentration (% v/v) and retention
time (min), as described by the empirical model, is
illustrated in a contour plot (Fig. 3). It should be noted
that this empirical model is based on the experimental
results and conditions prevailing in this study. Despite this
limitation, the model does provide a useful performance
estimation of similar biofilter systems. The focus of this
investigation, and subsequent model development, reflect
the extended residence times that would pertain to a
passive biofiltration system, rather than the relatively short
retention times typical of active biofiltration systems.
3.3. Removal efficiency
Methane removal efficiencies exceeding 70% were
obtained when the biofilter was subjected to a gas retention
time in excess of 50 min and methane concentrations up to
0.5% (v/v). From the contour plot it is evident that
the retention time had a major influence on the methane
removal efficiency. At retention times less than 30 min the
removal efficiency was significantly reduced. Similarly,
methane concentrations in excess of 0.5% (v/v) resulted in
reduced removal efficiencies regardless of retention time.
These results could possibly be attributed to a combination of
mass transfer limitations, due to the low solubility of
methane, and rate-limiting biodegradation capability within
the biofilm.
In this study methane oxidation capacity was allowed to
develop in a natural support medium, composted pine bark,
after prolonged exposure to methane. By comparison two
previous methane biofiltration investigations made use of
sterile biofilter support media, onto which specifically
cultured methane oxidizing bacteria, Methylomonas fodinarum and Methylomonas methanica, were immobilized [3,
4]. Sly et al., considered methane concentrations in the
range 0.25 1.0% at reactor residence times of 5 29 min in
a flow-through system, while Apel et al. considered
concentrations in the range 0 50.9% and residence times
of 0 7 h in a recirculating reactor system [3,4].
A comparison of the limited range of experimental

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

1363

Fig. 2. Effect of retention time on methane removal efficiency at methane concentrations of (a) 0.1%, (b) 0.3%, (c) 0.5%, (d) 1.0%, and (e) 2.5%.

condition overlap between this study and those of previous

investigations indicates an, expected, lower removal
efficiency of the composted pine bark biofilter compared
to specifically inoculated reactors. The most applicable
direct comparison is with that of Sly et al., at a residence
time of 20 min and 1% methane, where a 90% removal
efficiency was achieved compared to the 40% removal
efficiency obtained at similar conditions in this
investigation.

Table 1
Value parameters a and b (Eq. (1)) for various methane concentrations
Methane concentration (% v/v)

0.1
0.3
0.5
1.0
2.5

80.97
76.00
75.50
64.00
62.27

0.0688
0.0553
0.0450
0.0315
0.0200

1364

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

Fig. 3. Contour plot generated using the described empirical model (Data represents methane removal efficiencies for the outlet).

4. Conclusions

Acknowledgements

It can be concluded that composted pine bark may be

successfully utilised as a biofiltration support medium in
worked
underground
coal
mining
chambers.
Certain limitations would, however, apply to the potential
application of such biofiltration systems. Due to the
lengthy gas residence times required for effective
methane oxidation, such biofilters would best be operated
as passive air flow reactors rather than conventional
active blowing or drawing of gas flow through the
biofiltration bed. Although such passive biofiltration
systems would exhibit more efficient removal efficiencies
in the low range of methane concentrations, i.e. , 0.5%
(v/v), significant removal efficiencies (up to 50%) may be
achieved for methane concentrations approaching the
lower limit of the explosive range. This effect may,
therefore, prevent the escalation of methane concentrations in underground mining chambers, thus preventing
such concentration from reaching the lower explosive
limit. Further investigations would require pilot testing
and in situ monitoring in order to assess the practical
application potential of such biofiltration systems over a
prolonged period of time.

This research was sponsored by BHP Billiton and a

Technology for Human Resources in Industry Programme
(THRIP) research grant, South Africa. The authors thank
Prof Eric Senior, Dr Richard Daneel (University of Natal)
and Peet Jansen van Rensburg (Potchefstroom University for
Christian Higher Education), for their valuable assistance.

Appendix A
The following symbols were used:
a; b
equation fitting parameters
C
methane concentration (% v/v)
RE
removal efficiency (% of inlet concentration)
T
retention time (min)

References
[1] Windholz M, Budavari S, Blumetti RF, Ottenbein ES. The Merck
Index, 10th ed. ; 1983. Merck, Rahway, NJ, pp. 852853.

C.A. du Plessis et al. / Fuel 82 (2003) 13591365

[2] Johnson PW, Noval T, White DJ, Stevenson JW, Mills RA, Lasseter
EL, Boyer CM. IEEE Trans Ind Appl 1998;34(2):399 405.
[3] Apel WA, Dugan PR, Wiebe MR. Use of methanotrophic bacteria in
gas phase bioreactors to abate methane in coal mine atmospheres. Fuel
1991;70:1001 3.
[4] Sly LI, Bryant LJ, Cox JM, Anderson JM. Development of a biofilter
for the removal of methane from coal mine ventilation atmospheres.
Appl Microbiol Biotechnol 1993;39:4004.
[5] Devinny JS, Deshusses MA, Webster TS. Biofiltration for air
pollution control. Florida: CRC Press; 1999.
[6] Zehnder AJB, Brock TD. Methane formation and methane
oxidation by methanogenic bacteria. J Bacteriol 1979;137(1):
420 32.
[7] Anthony C. Bacterial oxidation of methane and methanol. In: Rose
AH, Tempest DW, editors. Advances in microbial physiology, Vol.
27. London: Academic Press; 1986. p. 113210.
[8] Hanson RS, Hanson TE. Methanotrophic bacteria. Microbiol Rev
1996;60(2):43971.
[9] Metcalf and Eddy Inc, In: Tchobanoglous G, Burton FL, editors.
Wastewater Engineering: Treatment, Disposal and Reuse, 3rd ed.
New York: McGraw-Hill; 1991. p. 1256.
[10] Patel RN, Hou CT, Laskin AI, Felix A, Derelanko P. Microbial
oxidation of gaseous hydrocarbons. II Hydroxylation of alkanes
and epoxidation of alkenes by cell-free particulate fractions
of methane-utilizing bacteria. J Bacteriol 1979;139(2):6759.
[11] Packer L, Rimbach G, Virgili F. Antioxidant activity and
biologic properties of a procyanidin-rich extract from pine (Pinus
maritima) bark, pycnogenol. Free Radic Biol Med 1999;27(5-6):
70424.
[12] Virgili F, Pagana G, Bourne L, Rimbach G, Natella F, Rice-Evans
C, Packer L. Ferulic acid excretion as a marker of consumption of

[13]
[14]

[15]

[16]

[17]
[18]

[19]

[20]

[21]

[22]

1365

a French maritime pine (Pinus maritima) bark extract. Free Radic

Biol Med 2000;28(8):124956.
Strauss JM, du Plessis CA, Riedel K-HJ. Empirical model for
biofiltration of toluene. J Environ Engng 2000;126(7):644 8.
du Plessis CA, Strauss JM, Riedel K-HJ. BTEX catabolism
interactions in a toluene-acclimatized biofilter. Appl Microbiol
Biotechnol 2001;55:1228.
Dunfield PF, Conrad R. Starvation alters the apparent half-saturation
constant for methane in the Type II methanotroph Methylocystis strain
LR1. Appl Environ Microbiol 2000;66(9):41368.
Wallace L, Paterson A, McCarthy A, Raeder U, Ramsey L,
MacDonald M, Haylock R, Broda P. The problem of lignin
biodegradation. Biochem Soc Symp 1983;48:87 95.
Vicuna R. Ligninolysis. A very peculiar microbial process. Mol
Biotechnol 2000;4(2):1736.
Jensen S, Ovreas L, Daae FL, Torsvik V. Diversity in methane
enrichments from agricultural soil revealed by DGGE separation of
PCR amplified 16S rDNA fragments. FEMS Microbiol Ecol 1998;26:
1726.
Madrid VM, Taylor GT, Scranton MI, Chistoserdov AY. Phylogenetic
diversity of bacterial and archaeal communities in the anoxic zone of
the Cariaco Basin. Appl Environ Microbiol 2001;67(4):1663 74.
Plumb JJ, Bell J, Stuckey DC. Microbial populations associated with
treatment of an industrial dye effluent in an anaerobic baffled reactor.
Appl Environ Microbiol 2001;67(7):322635.
Hrsak D, Grbic-Galic D. Biodegradation of linear alkylbenzenesulphonates (LAS) by mixed methanotrophic-heterotrophic cultures.
J Appl Bacteriol 1995;78(5):48794.
van Bodegom P, Stams F, Mellema L, Boeke S, Lefelaar P. Methane
oxidation and the competition for oxygen in the rice rhizosphere. Appl
Environ Microbiol 2001;67(8):3586 97.