International Journal of Coal Geology 76 (2008) 8697

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International Journal of Coal Geology

j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / i j c o a l g e o

Biodegradation of sedimentary organic matter associated with coalbed methane in the Powder River and San Juan Basins, U.S.A.
Michael Formolo a,, Anna Martini b, Steven Petsch a
a b

Department of Geosciences, University of Massachusetts, Amherst, MA, 01003-9297, USA Department of Geology, Amherst College, Amherst, MA, 01002, USA

A R T I C L E

I N F O

A B S T R A C T
The Powder River Basin and San Juan Basin, U.S.A., are two of the most productive coalbed methane reserves in the world. Of particular interest is the microbial biodegradation of coal beds associated with this natural gas production. Biogenic methane production is indicated as a signicant component to the total gas resources in the San Juan Basin, and as the nearly sole source for the shallow coals of the Powder River Basin. Molecular and isotopic signatures indicate a microbial origin for the gas. Geochemical characteristics of formation waters, such as elevated alkalinity and 13C-enriched dissolved inorganic carbon (DIC), further support extensive microbial degradation of coal organic matter associated with methanogenesis. Extractable organic matter isolated from coals in both basins point to patterns of hydrocarbon biodegradation in coals restricted to specic depths. To some extent, biodegradation patterns are similar to those observed in methanogenic, biodegraded black shales of the mid-continent of the United States. Specically, both coals and shales exhibit near-quantitative removal of straight-chain and acyclic isoprenoid hydrocarbons. However, loss of aromatic hydrocarbons in the coals proceeds prior to the extensive removal of the saturated hydrocarbons, in contrast to what is conventionally observed in biodegraded petroleum systems or in black shales. In addition, previous thermal maturation histories in both the Fruitland and Fort Union coalbed methane systems have little impact on more recent hydrocarbon biodegradation. Instead, localized hydrologic conditions and subsurface geology likely play important roles in controlling the extents of biodegradation and methanogenesis. These results suggest that biodegradation of hydrocarbons coupled with methanogenesis may develop regardless of organic matter source across a range of inherited thermal maturities. 2008 Elsevier B.V. All rights reserved.

Article history: Received 1 August 2007 Received in revised form 29 February 2008 Accepted 12 March 2008 Available online 29 March 2008 Keywords: San Juan Basin Powder River Basin Biodegradation Sedimentary organic matter Coalbed methane

1. Introduction Economic production of natural gas from coal beds began in the United States in the 1930s, but in recent decades coalbed gas development has increased dramatically and now accounts for approximately 9.3% of the annual production of dry gas in the United States (Ayers, 2002; Energy Information Agency, 20061). Two prolic coalbed gas reservoirs in the United States are the Cretaceous Fruitland Formation of the San Juan Basin and the Paleocene Fort Union Formation of the Powder River Basin. Understanding the subsurface processes leading to the formation of coalbed methane is important for the present production and future exploration of these deposits. Two mechanisms form coalbed gas: biological processes (i.e. methanogenesis) or geologic processes (i.e. thermogenic gas production). In

Corresponding author. Current address: Department of Biogeochemistry, MaxPlanck-Institute for Marine Microbiology, Celsiusstr. 1, D-28359 Bremen, Germany. Tel.: +49 421 2028 655. E-mail address: mformolo@mpi-bremen.de (M. Formolo). 1 http://www.eia.doe.gov/. 0166-5162/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.coal.2008.03.005

this study, we examine coalbed gas reserves that are biogenic or mixed biogenic/thermogenic in origin. Microbial methanogenesis is the main source for gas in the Powder River Basin (Ayers, 1986; Gorody, 1999; Flores et al., 2008-this volume) and a signicant addition to the thermogenic gas found in the San Juan Basin (Rice et al., 1989; Clayton et al., 1991; Kaiser et al., 1991; Scott et al., 1991; Michael et al., 1993; Rice, 1993; Scott et al., 1994; Zhou et al., 2005). The dominant coalbed methane producing formation in the Powder River Basin is the Paleocene Fort Union Formation. The Fort Union Formation consists of the Tullock, Lebo Shale, and Tongue River Members (Fig. 1A). Lithologically the Tullock Member contains evidence of deposition within uvial environments; the Member is dominated by sandstone, mudstone, shale, and coal (Flores and Etheridge, 1985; Flores et al., 2008-this volume). Overlying the Tullock Member is the Lebo Shale Member, which contains abundant shale, mudstones, siltstones, and sandstones, while containing minor amounts of coal. The dominant coal-bearing Member is the Tongue River Member. The Tongue River contains 32 coals seams that range in thickness from a meter to tens of meters (Ayers, 1986; Montgomery, 1999). High-resolution lithological and stratigraphic information for

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87

Fig. 1. Generalized stratigraphic column for the Powder River Basin (A) and San Juan Basin (B).

the Powder River Basin is available in Ayers and Kaiser (1994), Tyler et al. (1995), and Stricker et al. (2007). Recent data from closely spaced CBM wells show Tongue River coal beds to be very variable laterally from a few meters thick to more than 60-m thick within several kilometers (Flores, 2004). The principal methane producing coal formations in the San Juan Basin are contained in the Upper Cretaceous Fruitland Formation (Fig. 1B). The Fruitland Formation consists of interbedded coal, sandstone, and shales, with some coalbeds reaching 30 m in thickness (Fassett, 1988; Ayers and Kaiser, 1994; Pashin, 1998; Craigg, 2001, Brister and Hoffman, 2002; Lucas et al., 2006). The coal-bearing Fruitland Formation formed in coastal plain settings associated with the Late Cretaceous Western Interior seaway (Fassett and Hinds, 1971; Ayers et al., 1991). Ayers and Ambrose (1990), Ayers et al. (1991), Robinson Roberts and McCabe (1992), and Pashin (1998) provide highresolution stratigraphic cross-sections of the San Juan Basin and the Fruitland Formation. In both Powder River and San Juan Basins, stimulation of biogenic gas generation has been linked to incursion of meteoric water (Kaiser et al., 1991; Scott et al., 1991; Rice, 1993; Scott et al., 1994; Montgomery, 1999; Zhou et al., 2005). Regional hydrology may be an important mechanism stimulating biogenic gas production in the San Juan and Powder Rivers Basins (Flores et al., 2008-this volume), similar to the shale-gas reserves in the Michigan Basin (Martini et al., 1996, 1998; McIntosh et al., 2004). While geochemical and isotopic indicators establish that gas in these basins contains microbial methane (Rice et al., 1989; Rice, 1993; Scott et al., 1994; Gorody, 1999), little is understood regarding consumption of coal organic matter leading to methanogenesis or the signatures of hydrocarbon biodegradation associated with biological methane generation. In this study, we examine solvent-extractable organic matter isolated from samples of the Fruitland coal (San Juan Basin) and Fort Union coal (Powder River Basin) to determine the relative abundance of saturated and aromatic hydrocarbons in these coals, and to evaluate the source, maturity and biodegradation of coals associated with coalbed methanogenesis in the Powder River and San Juan Basins. Calculation of biodegradation indices for hydrocarbon compound classes allows us to describe the molecular signatures of hydrocarbon biodegradation in coalbed methane systems. By establishing similar thermal histories and organic matter sources any

differences in our biodegradation ratios are the result of biodegradation and not source or thermal inuences. 2. Methods Coal samples from the Powder River and San Juan Basin (Fig. 2) were collected from the USGS Core Repository in Denver, Colorado. Sample locations and USGS library reference numbers are listed in Table 1. The Fort Union Formation and the Fruitland Formation were targeted in the Powder River and San Juan Basins, respectively, as these are the two dominant coal coal-bearing formations associated with gas production. During sub-sampling of the cores, particular attention was given to trying to select the least weathered coals by sampling from the center of the core when possible. Samples were chosen to reect a range of burial depths and thermal maturities across each basin. Total organic carbon (TOC) concentrations were determined using a Costech ECS140 Elemental Analyzer. Bulk organic matter 13C compositions were determined by online combustion using a Costech ECS140 EA interfaced to a Thermo Delta 5 isotope-ratio mass spectrometer. Analyses were run in triplicate and are reported relative to the Vienna PDB in standard per mil () notation with a precision of 0.3. Vitrinite reectance (Ro) values in percent (%) were obtained on polished, vacuum epoxy xed, chips of coal from seven samples representing six different well locations. The surface was examined for vitrinite using a petrographic microscope and ten counts were made for 1219 sites per chip and values are reported as averages s.d. (1). Though the maturity rank of the coals are low, vitrinite reectance data can provide adequate determination of maturity (Cannico et al., 2004; Skorov et al., 2005. To isolate extractable organic matter, 6 to 10 g of crushed Fort Union and Fruitland coals were solvent extracted in 3:1 dichloromethane (DCM): methanol (MeOH) for 72 h in a Soxhlet apparatus. Total extractable organic matter (EOM) was determined by gravimetric measurements following solvent extraction. The extracts were separated on silica gel into aliphatic (F1), aromatic (F2), and polar fractions (F3), by the successive elution with hexane, hexane: dichloromethane (1:1), and dichloromethane:methanol (1:1), respectively (Brocks et al., 2005). Fractions were dried under a puried N2 stream at room temperature.

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Fig. 2. Map of the sample locations. Numbers refer to depth (m) of sample. (A) The Powder River Basin; (B) The San Juan Basin.

Fractions F1 and F2 were redissolved in hexane and analyzed on a HP 6890 gas chromatograph (GC) interfaced to a HP 5973 massselective detector, using helium as a carrier gas. The GC was equipped with a split/splitless injector and a 30 m Hewlett-Packard HP-5MS fused silica capillary column (0.25 mm i.d., 0.25 m lm thickness). For F1, the GC temperature program was held isothermal at 60 C for 1.5 min followed by an increase of 20 C/min until 130 C and then a 2 C/min increase to 300 C, then held isothermal for 10.0 min. For F2, the GC temperature program was initially set at 60 C for 2.0 min and followed by an increase of 20 C/min until 150 C and then followed by a 2 C/min increase until 320 C, then held

isothermal for 10 min. For both analyses, the mass-spectrometer source was operated at 250 C in EI-mode at 70 eV ionization energy. Identication of compounds was based on interpretation of gas chromatograph-mass spectra and comparison with published mass spectra and retention times (Figs. 3 and 4). Quantication of individual compounds was achieved using extracted ion chromatograms and peak area integration, using m / z = 57 (alkanes and acyclic isoprenoids), m / z = 191 (hopanes), m / z = 128, 156, 170, 184 (alkylated naphthalenes), and m / z = 178, 192, 206, 220, 234 (alkylated phenanthrenes) (Peters and Moldowan, 1993; Fisher et al., 1996, 1998; Ahmed et al. 1999; Ahmed and Smith, 2001; Peters et al., 2005).

Table 1 Sample depth, organic geochemistry, source indicators, and 13C()V-PDB of the Fort Union and Fruitland coals Formation Latitude (N)/Longitude (W) USGS reference # S057 D395 D395 C590 Depth (m) 132 750 760 1190 Average TOCa wt.% 33.94 63.03 60.15 70.86 57.00 EOMb (mg)/g TOC 101.3 111.19 109.18 101.98 13C ()V-PDB 26.2 26.6 26.2 26.7 Average CPIc 2.20 1.14 1.15 1.05 OEP(17)d 1.65 1.98 6.47 2.14 OEP(29)d 3.68 1.85 1.62 1.72 Pr/Phe 9.20 7.72 6.64

San Juan Basin Fruitland 362018.24/1073817.16 Fruitland 365034.80/1074355.20 Fruitland 365034.80/1074355.20 Fruitland 364054.48/1071223.40

26.8 26.53 0.3

26.8

Powder River Basin Fort Union 453036.72/1065239.72 Fort Union 453036.72/1065239.72 Fort Union 446015.48/1053320.88 Fort Union 446015.48/1053320.88 Fort Union 43516.12/1052943.80 Fort Union 444541.04/106518.96
a b c d e

S939 S939 S739 S739 S352 S794

49 146 223 224 266 429 Average

57.49 59.22 58.08 63.36 57.84 59.06 59.18

60.54 202.74 37.3 74.45 104.6 57.64

25.8 24.9 24.8 24.1 24.1 22.7 Average

2.46 2.17 2.83 0.94 2.44 2.82 24.4 1.1

2.15 1.21 1.21 1.29 1.35 1.06

3.87 3.12 3.67 3.64 3.84

4.17 4.95 4.26 2.33 5.98 5.45

TOC = total organic carbon. EOM = extractable organic matter. CPI = 2(C23 + C25 + C27 + C29/[C22 + 2(C24 + C26 + C28) + C30]. OEPx = [C(x 2) + 6Cx + C(x + 2)]/[4C(X 1) + 4C(x + 1)]. Pr/Ph = pristane/phytane.

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Fig. 3. Representative total ion current [TIC] chromatograms obtained from GC-MS analysis of saturated and aromatic hydrocarbon fractions isolated from selected depths of the Fruitland Coal, San Juan Basin. UCM = unresolved complex mixture.

Source and maturity indices were calculated following Peters et al. (2005) and references therein. Source indices include carbonpreference index (CPI) and odd-over-even predominance (OEP). Terrestrial land plants contribute leaf waxes comprising odd carbon number n-alkanes, and organic matter sources dominated by terrestrial land plants will result in elevated (N 1) CPI and OEP ratios (Tissot and Welte, 1984). CPI was integrated over the entire range of available n-alkanes, while OEP ratios were separated into short (OEP17) and long (OEP29) chain n-alkanes to address the oddover-even preference in specic n-alkane chain lengths. Pristane/ Phytane (Pr/Ph) ratios were also measured to address depositional conditions. In addition to the vitrinite reectance analyses, molecular maturity indices include hopane stereoisomer ratios, specically the ratio of C27 17(H)-trisnorhopane (Tm) to C27 18(H)-trisnorhopane II (Ts) (Siefert and Moldowan, 1978) and the isomerization of C31C34 17(H)21(H)-homohopanes. Generally, these ratios increase with increasing thermal maturity, (Ensminger et al., 1974; Siefert and Moldowan, 1978; Peters and Moldowan, 1993; Peters et al., 2005). The isomerization of C31-17(H)21(H)-homohopanes is governed by the rates of generation and thermal degradation of these hopanes (Farrimond et al., 1998), however, in these relatively immature

coals it is likely that minimal thermal degradation has occurred therefore maintaining the integrity of the molecular signatures. In addition, due to the inuences of source facies on Ts and Tm ratios it was necessary to integrate multiple maturity indicators to adequately determine the thermal maturity of the coals. Utilizing the biodegradation scheme of Peters and Moldowan (1993) and Peters et al. (2005) it was determined that the most bioresistant compounds present in our samples are the C31-homohopanes. Biodegradation indices were calculated based on loss of the more susceptible compounds relative to the sum of the more resistant C31-17(H)-21(H) 22S and 22R hopanes as follows: Biodegradation Index = X X C31 hopanes= compounds of interest: 1

For our calculations, greater values of the biodegradation index indicate increased relative removal of the compound(s) of interest, i.e. greater biodegradation. Biodegradation was evaluated for the following: nC15nC19 alkanes, nC20nC24 alkanes, nC25nC30 alkanes, three acyclic isoprenoids (norpristane, pristane, phytane), nC18 nC27

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Fig. 4. Representative total ion current [TIC] chromatograms obtained from GC-MS analysis of saturated and aromatic hydrocarbon fractions isolated from selected depths of the Fort Union Coal, Powder River Basin. UCM = unresolved complex mixture.

alkylcyclohexanes, methylnaphthalene (MN), dimethylnaphthalene (DMN), trimethylnaphthalene (TrMN), tetramethylnaphthalene (TeMN), phenanthrene (P), methylphenanthrene (MP), dimethylphenanthrene (DMP), trimethylphenanthrene (TrMP), and tetramethylphenanthrene (TeMP). Since the possibility of comparing different compound classes, e.g. saturated versus aromatic hydrocarbons, may be subjected to biases from potential differences in the response factors of mass fragment ions we also assessed the biodegradation of aromatic hydrocarbons by taking the ratios of less-susceptible aromatic compound classes over more-susceptible compound classes. This approach is similar to the approach by Ahmed et al. (1999) where the authors compared ratios of compounds that had varying susceptibility to biodegradation. The additional aromatic biodegradation index is: Aromatic X Index X less susceptible compounds= more susceptible compoundss 2 And similar to the Biodegradation Index, increased values indicate the preferential removal of the more easily biodegraded compound.

3. Results 3.1. Source parameters Results for the Fort Union coal and Fruitland coal are tabulated in Table 1. Fort Union TOC values range from 57.49 to 63.36 wt.% (average = 59.18 wt.%, n = 7). Fruitland TOC values range from 33.94 to 70.86 wt.% (average = 57.00 wt.%, n = 4), with the lowest concentration measured at 132-m depth. Total EOM (mg)/g TOC values in the Fort Union coal range are highly variable and range from 37.3 to 202.74 mg/g TOC. Total EOM (mg)/g TOC values in the Fruitland coal exhibit little variability and range from 101.3 to 111.19 mg/g TOC. Bulk organic matter 13C values for the Fort Union coal range from 22.7 to 25.8 (average = 24.4 1.1). Bulk organic matter 13C values for the Fruitland have an average value of 26.5 0.3, and a range of 26.2 to 26.7. Biomarker indices recording the source input include CPI, OEP17, and OEP29. CPI values for the Fort Union coal range between 2.17 and 2.83, with an anomalous value of 0.94 at 224 m. Fort Union coal OEP17 values range between 1.06 and 1.35, except for the shallowest depth (49 m), which has a value of 2.15. Fort Union coal OEP29 values range

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from 3.12 to 3.87. Pr/Ph ratios throughout the core and range from 4.17 to 5.98, with one anomalous value of 2.33 at 224 m. A CPI value of 2.20 was calculated for the shallowest Fruitland coal sample (132 m), while the other three depths have values between 1.05 and 1.15. Fruitland coal OEP17 values are similar to those from the Fort Union coal, and range between 1.65 to 2.14; however, at a depth of 760 m the value increases to 6.47. The Fruitland coal OEP29 range from 1.62 to 3.68 and the Fruitland Pr/Ph ratios range from 6.64 to 9.20. 3.2. Maturity indices Vitrinite reectance and molecular maturity indices for the Fort Union and Fruitland coals are tabulated in Table 2. Vitrinite reectance (Ro) data from the Fort Union coals range from 0.45 to 0.53% (average = 0.49%). Ts/(Ts + Tm) ratios in the Fort Union coal are 0.20 to 0.25 in the shallowest four samples and decrease to 0.13 and 0.16 in the deepest two samples. Fort Union coal C31 22S/22R ratios are extremely variable and range from 0.03 to 0.82. Fort Union coal C32 maturity ratios are between 0.56 and 0.74 with an outlying value of 0.02 at 224 m. The C33 ratios determined in the Fort Union coal are 0.43, 0.02, 0.03, and 0.01, with increasing depth. Fort Union coal C34 hopane ratios ranged from 0.29 to 0.68. Ro data was calculated for two of the Fruitland Formation coals: Rovalues are 0.55 and 0.82%. Ts/(Ts + Tm) ratios in the Fruitland coal range from 0.07 to 0.94. The Fruitland coal C31 22S/22R ratio is 0.26 at 132 m and range between 0.580.59 in deeper samples, similar to results reported by Michael et al. (1993). Fruitland coal C32 ratios are similar to the C31 ratios, equaling 0.13 at 132 m and ranging from 0.58 to 0.59 in deeper samples. The Fruitland coal C33 ratio is 0.52 at 132 m, then ranges from 0.580.60 at depth. The C34 hopane ratio could not be determined for the shallowest sample of Fruitland coal but ranged from 0.60 to 0.61 in the samples from greater depth. 3.3. Biodegradation indices To assess biodegradation, the abundance of compound(s) of interest relative to the summed abundance of the C3117(H)21(H)hopanes 22S and 22R isomers was calculated. C3117(H)21(H)hopanes were chosen as non-degraded reference compounds as these are known to be highly resistant to biodegradation (Peters and Moldowan, 1993; Peters et al., 2005). Similar source inputs and thermal histories suggest that the relative inputs and generation or thermal degradation of the hopane isomers should be consistent in the samples from their respective basins. The pristane/nC17 and phytane/nC18 ratios were also calculated as parameters for degrada-

Fig. 5. Comparison of source and maturity parameters versus biodegradation indices for the Fruitland and Fort Union Formations coals. (A) Comparison of organic matter source revealed by carbon preference index values (CPI) versus hydrocarbon biodegradation indices for Fruitland and Fort Union Coals. R2 values from linear regression analysis are provided in gure legend. (B) Comparison of Ts/(Ts + Tm) maturity index values versus hydrocarbon biodegradation indices for Fruitland and Fort Union Coals. R2 values from linear regression analysis are provided in gure legend.

Table 2 Vitrinite reectance and hopane maturity biomarker indices Maturity indices Formation Depth (m) Ro (ave. s.d.) Ts/(Ts + Tm)a S/(S + R)b C31 Fruitland Fruitland Fruitland Fruitland Fort Union Fort Union Fort Union Fort Union Fort Union Fort Union 132 750 760 1190 49 146 223 224 266 429 0.55 0.04 0.82 0.06 0.37 0.07 0.94 0.26 0.25 0.25 0.25 0.20 0.13 0.16 0.26 0.59 0.59 0.58 0.82 0.05 0.06 0.03 0.79 0.08 C32 0.13 0.59 0.59 0.58 0.58 0.74 0.50 0.02 0.56 C33 0.52 0.60 0.58 0.58 0.43 0.03 0.02 0.03 0.01 C34 0.61 0.60 0.61 0.29 0.68 0.46 0.47 0.45

0.53 0.04 0.45 0.05 0.48 0.04 0.47 0.04 0.52 0.04

tion of methyl-branched versus straight-chain saturated hydrocarbons. These ratios can be used as biodegradation indicators if samples have similar sources and experienced similar thermal histories. Importantly, regression analysis revealed no discernible relationships between source or maturity parameters and biodegradation indices in either the Fort Union or Fruitland coals (Fig. 5). In addition to the C31-hopane biodegradation index, the relative biodegradation of aromatic hydrocarbons was determined by comparing aromatic compounds more susceptible to biodegradation to aromatic compounds that are less susceptible from the same compound classes. This calculation was performed to assure that biases from variations in the mass fragment responses between saturated and aromatic hydrocarbons would be not be a factor. 3.3.1. Saturated hydrocarbons Biodegradation indices for saturated hydrocarbons are tabulated in Table 3. Elevated biodegradation indices of n-alkanes and acyclic isoprenoids, specically norpristane, pristane, and phytane, in the Fruitland coal was calculated at 132 and 760 m depths (Fig. 6A), representing biodegradation at these depths. Values in the Fort Union

a Ts/(Ts + Tm) = C27 18(H)-trisnorhopane II (Ts)/(C27 18(H)-trisnorhopane II (Ts) + C27 17(H)-trisnorhopane(Tm)). b 22S/(22S + 22R) = 17 (H),21 (H)-22S homohopane/[17 (H),21 (H)-22S homohopane + 17(H),21(H)-22R homohopane].

92 Table 3 Biodegradation indices for saturated hydrocarbons

M. Formolo et al. / International Journal of Coal Geology 76 (2008) 8697

n-alkanes Formation Fruitland Fruitland Fruitland Fruitland Fort Union Fort Union Fort Union Fort Union Fort Union Fort Union
a b c d e f g

Depth (m) 132 750 760 1190 49 146 223 224 266 429

Pristane/nC17 6.84 2.39 10.83 0.71 2.62 7.46 0.94 0.95 1.69 2.86

Phytane/nC18 0.77 0.55 15.30 1.03 1.62 0.33 0.62 0.46 0.62

(nC15nC19)c 3.19 0.27 6.35 0.06 2.37 15.62 1.00 6.61 0.51 1.29

(nC20nC24)d 1.45 0.22 8.52 0.04 1.34 9.79 1.29 8.16 0.66 0.93

(nC25nC30)e 0.50 0.20 1.01 0.07 0.66 6.70 1.18 9.01 0.69 0.41

(acyclic isoprenoids)f 2.20 0.28 1.01 0.06 1.64 8.05 2.17 8.60 0.87 1.26

(n-alkyl-(CH))g 26.68 0.53 0.08

Pr/n-C17 = pristane/C17 n-alkane. Ph/n-C18 = phytane/C18 n-alkane. (nC15-nC19) = (C31-hopanes) / ((nC15nC19). (nC20nC24) = (C31-hopanes) / (nC20nC24). (nC25nC30) = (C31-hopanes) / (nC25nC30). (acyclic isoprenoids) = (C31-hopanes) / (norpristane + pristane + phytane). (n-alkyl-(CH)) = (C31-hopanes) / (n-alkylcyclohexanes).

coal are highest at depths of 146 and 224 m, indicating biodegradation located at these depths (Fig. 6B). The Fruitland coals exhibit a series of n-alkylcyclohexanes, specically nC18nC27 cyclohexanes. This series of n-alkylcyclohexanes were not observed in the Fort Union coal. In the Fruitland coals,

similar to both the n-alkanes and acyclic isoprenoids, the nalkylcyclohexanes show variations in the biodegradation index with depth (Table 3). At 132 m the value is 26.68, at 750 m the value is 0.53, and at 1302 m the value is 0.08. Biodegradation indices for saturated hydrocarbons exhibit minima for the deepest samples of both the Fort Union and Fruitland coals (429 m and 1190 m, respectively). 3.3.2. Aromatic hydrocarbons Variations in the distribution of the aromatic hydrocarbons also highlight the occurrence of biodegradation in these coals. Calculated biodegradation and aromatic indices for aromatic hydrocarbons are provided in Tables 4 and 5. In the Fruitland coal of the San Juan Basin, biodegradation indices calculated for the naphthalene and phenanthrene series show elevated biodegradation at 132, 750, and 760 m (Fig. 7A and B). The results and depth distributions from the aromatic index are similar to the results from the biodegradation index (Tables 4 and 5). It is apparent from these two calculated indices that there has been preferential removal of the more susceptible compounds. Interestingly, at a depth of 1190 m in the Fruitland there are extremely elevated TMN/DMN ratios, this could be an artifact of an increased thermal maturity. Values for naphthalenes suggest biodegradation throughout most of the Fort Union coal samples (Fig. 8A). There were no identiable MNs in the Fort Union coal and only minimal DMNs. Only three measurements could be made for the DMNs; however the biodegradation indices are elevated at 223 m compared to the deeper samples. The TrMN and TeMN show elevated biodegradation at 146 and 224 m, similar to the saturated hydrocarbons. Biodegradation indices calculated for Fort Union coal phenanthrenes show patterns similar to the naphthalenes (Fig. 8B). Phenanthrene biodegradation indices in the Fort Union are high for most of the sample locations, with the most elevated biodegradation indices at 146 and 224 m. As observed for saturated hydrocarbons, the biodegradation index for aromatic hydrocarbons exhibit minima for the deepest samples of both the Fort Union and Fruitland coals (429 m and 1190 m, respectively). The aromatic index, similar to the biodegradation index, indicates the preferential removal of the more susceptible compounds in both the naphthalene and phenanthrene compound classes (Table 5). However, at a depth of 429 m there appears to be elevated TMN/DMN and MP/P ratios, these may be the result of local heterogeneities or a variation in thermal maturity, though the latter is unlikely based upon other results. Though the two separate indices compare different compound classes they both suggest there is the preferential removal of the more

Fig. 6. Calculated biodegradation indices for the saturated hydrocarbons for the Fruitland Coal and Fort Union Coal. Short-chain n-alkanes = nC15nC19. Medium-chain n-alkanes = nC20nC24. Long-chain n-alkanes = nC25nC30. Acyclic isoprenoids include norpristane, pristane, and phytane.

M. Formolo et al. / International Journal of Coal Geology 76 (2008) 8697 Table 4 Biodegradation indices for aromatic hydrocarbons Naphthalenes Formation Fruitland Fruitland Fruitland Fruitland Fort Union Fort Union Fort Union Fort Union Fort Union Fort Union
a b c d e f g h i

93

Phenanthrenes DMN 1.34 13.73 2.76 1.51

b

Depth (m) 132 750 760 1190 49 146 223 224 266 429

MN 9.36

TrMN 43.59 10.29 1.26 0.52 7.48 8.75 0.57 14.31 0.22 0.10

TeMN 38.90 0.21 1.40 0.47 4.20 5.60 0.73 5.20 0.35 0.43

MPf 14.81 0.10 0.62 0.09 0.98 1.44 0.68 3.54 0.12

DMPg 9.62 0.07 0.41 0.06 2.23 1.08 0.38 1.34 0.09

TrMPh 17.35 0.11 0.59 0.10 1.75 0.83 3.08 0.25

TeMPi 41.19 0.45 4.09 0.39 15.18

63.92 0.22 46.18 0.34 11.53 15.41 2.37 13.08 0.90

MN = (C31-hopanes) / (methylnaphthalene). DMN = (C31-hopanes) / (dimethylnaphthalenes). TrMN = (C31-hopanes) / (trimethylnaphthalenes). TeMN = (C31-hopanes) / (tetramethylnaphthalenes). P = (C31-hopanes) / (phenanthrene). MP = (C31-hopanes) / (methylphenanthrene). DMP = (C31-hopanes) / (dimethylphenanthrenes). TrMP = (C31-hopanes) / (trimethylphenanthrenes). TeMP = (C31-hopanes) / (tetramethylphenanthrenes).

susceptible compounds when compared to a more bio-resistant compound from the same class. 4. Discussion 4.1. Biodegradation of coal associated with coalbed methane Microbial methanogenesis is the main source for gas in the Powder River Basin (Ayers, 1986; Gorody, 1999; Flores et al., 2008-this volume) and a signicant addition to the thermogenic gas found in the San Juan Basin (Clayton et al., 1991; Kaiser et al., 1991; Michael et al., 1993; Scott et al., 1994; Zhou et al., 2005). Though the evidence and mechanisms involved in the production of coalbed methane have been intensively studied, few studies have focused on the signatures of organic matter biodegradation preserved in the coals (Michael et al., 1993; Curry et al., 1994; Ahmed et al., 1999; Ahmed and Smith, 2001). Qualitative observations, specically the generation of the unresolved complex mixture, of the total ion chromatograph of the saturated hydrocarbons indicate that both the Fruitland coal in the San Juan Basin and the Fort Union coal in the Powder River Basin have experienced some degree of biodegradation (Figs. 3 and 4). These chromatographs are dominated by a bimodal distribution and an increase in the UCM, qualitative indications of biodegradation. Unimodal distributions in

coals would be more indicative of increased thermal maturity (Ahmed et al., 1999). These characteristics are further indication that these observations are not the result of thermal maturity variations and suggest biodegradation.

Table 5 Aromatic indices for naphthalene and phenanthrene compound classes Formation Fruitland Fruitland Fruitland Fruitland Fort Union Fort Union Fort Union Fort Union Fort Union Fort Union
a b c d e f

Depth (m) 132 750 760 1190 49 146 223 224 266 429

TMN/ DMNa 7.00 58.89 24.10 12.41 14.80

TeMN/ TMNb 1.12 0.47 0.90 1.13 1.78 1.56 0.78 2.75 0.63 0.24

MP/ Pc 4.32 2.30 74.13 3.69 11.78 10.68 3.50 3.69 1.36 7.23

DMP/ MPd 1.54 1.30 1.50 1.54 0.44 1.34 1.77 2.64 0.83 1.40

TrMP/ DMPe 0.55 0.69 0.71 0.63 0.61 0.46 0.44 0.35

TeMP/ TrMPf 0.42 0.24 0.14 0.25 0.20 Fig. 7. Calculated biodegradation indices for the aromatic hydrocarbons from the Fruitland Coal, San Juan Basin. MN = methylnaphthalenes. DMN = dimethylnaphthalenes. TrMN = trimethylnaphthalenes. TeMN = tetramethylnaphthalenes.P = pheneanthrenes. MP = methylphenanthrenes. DMP = dimethylphenanthrenes. TrMP = trimethylphenanthrenes. TeMP = tetramethylphenanthrenes.

TMN/DMN = Trimethylnaphthalene/Dimethylnaphthalene. TeMN/TMN = Tetramethylnaphthalene/Trimethylnaphthalene. MP/P = Methylphenanthrene/Phenanthrene. DMP/MP = Dimethylphenanthrene/Methylphenanthrene. TrMP/DMP = Trimethyylnaphthalene/Dimethylnaphthalene. TeMP/TrMP = Tetramethylnaphthalene/Trimethylnaphthalene.

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Inputs from the waxes of terrestrial land plants result in CPI and OEP values greater than 1.0, or an odd-over-even predominance (Bray and Evans, 1961; Eglington and Hamilton, 1967; Scalan and Smith, 1970; Tissot and Welte, 1984). The CPI and OEP(29) for both the Fort Union and Fruitland coals suggest a similar input of long chain nalkanes derived from terrestrial higher plants, with modication due to post-depositional maturation and biodegradation. 4.3. Thermal maturity The rank of the Fort Union coals fall into the lignite to subbituminous A over the entire basin (Law et al., 1991; Rice, 1993; Stricker et al., 2007, Flores et al., 2008-this volume), making these the lowest grade coals in the United States to actively be exploited as a natural gas reserve. Stricker et al. (2007) shows an increase in coal rank (subbituminous C to A) from the margins to the central part of the Powder River Basin. The coals in the San Juan Basin are higher in rank than those of the Powder River Basin. The Fruitland coals increase in rank from the southwest to northeast portion of the basin, from subbituminous C to medium- and low-volatile bituminous, respectively (Rice, 1983; Rice et al., 1989; Law, 1992; Rice, 1993; Scott et al., 1994). Although these ranks of coals are well established, determining the individual thermal maturities of the coals analyzed in this study is important prior to accessing the level of biodegradation. Extremely high and anomalous maturities could inuence the abundance of aliphatic and aromatic hydrocarbons and could lead to false determinations of biodegradation levels. All maturity indicators are listed in Table 2. Fort Union and Fruitland coals Ro values differ as expected. Fort Union coal Ro values are indicative of immature to transitional maturities and the Fruitland Ro values suggest transitional to mature levels of thermal maturity. These vitrinite reectance data are consistent with the thermal histories of the Powder River and San Juan Basins (Nuccio, 1990; Michael et al., 1993; Rice, 1993; Stricker et al., 2007; Flores et al., 2008-this volume). As expected, Ts/(Ts + Tm) maturity ratios also differ. During increasing thermal maturation, Tm is less stable and will decrease in relative concentration compared to Ts (Waples and Machihara, 1991), thereby increasing the ratio. Because organic matter source inputs are similar within the respective basins, these ratios record maturity or other changes, such as biodegradation. The San Juan Basin Fruitland coals show elevated Ts/(Ts + Tm) ratios for three of the four coals. However, there is no apparent relationship between depth and increasing maturity, potentially indicating that different portions of the basin have experienced variable localized thermal histories. In contrast to the Fruitland coals, Ts/(Ts + Tm) ratios in the Powder River Basin Fort Union coal are uniform and low in all the samples. These indicate the low-grade nature of the coal, consistent with the inferred geologic history of the Powder River Basin (Nuccio, 1990; Rice, 1993; Stricker et al., 2007; Flores et al., 2008-this volume). To more accurately constrain the thermal history of these coals the C31C34 17 (H) 21(H)homohopane 22S/(22S + 22R) ratios were determined. These apparent isomerization ratios are controlled by relative rates of generation and destruction during thermal maturation (Bishop and Abbot, 1993; Farrimond et al., 1998), typically reaching equilibrium around values of 0.570.62 (Seifert and Moldowan, 1986). Therefore, assuming that the relative rates of generation and destruction are of these hopane isomers are similar, these ratios provide an indication of the thermal history of the sedimentary organic matter. For samples from within basins subjected to similar thermal histories and sources of organic matter, the relative degrees of isomerization should be comparable. These ratios may differ slightly but should be in relative agreement if an external process, such as source variation or biodegradation, does not alter the respective hopanes. C31C34 homohopane ratios for the Fruitland coal are all approximately at equilibrium except for the shallowest coal. The shallowest Fruitland coal (132 m) has a C31 and C32 hopane ratio

Fig. 8. Biodegradation indices for the aromatic hydrocarbons for the Fort Union Coal, Powder River Basin. DMN=dimethylnaphthalenes. TrMN=trimethylnaphthalenes. TeMN=tetramethylnaphthalenes. P =pheneanthrenes. MP=methylphenanthrenes. DMP=dimethylphenanthrenes. TrMP=trimethylphenanthrenes. TeMP=tetramethylphenanthrenes.

Biodegradation in petroleum reservoirs (Head et al., 2003; references therein) has been well established; however, the anaerobic biodegradation of sedimentary organic matter associated with methanogenesis and coalbed methane are less well known. Recently, a study by Formolo et al. (2008) demonstrated that biodegradation of shale hosted organic matter, leading to the production of biogenic gas, in the Antrim Formation is similar to what is observed for petroleum reservoirs, though few studies have addressed the biodegradation of coals associated with coalbed methane (Michael et al., 1993; Ahmed et al., 1999; Ahmed and Smith, 2001). To address biodegradation signatures, it is necessary to assess the source inputs and thermal histories of these basins so we can determine that any measurable differences are due to external processes such as biodegradation. 4.2. Source of organic matter The 13C isotopic composition of the Fruitland coals fall within a relatively narrow range, 25.9 to 26.8 (Table 1), indicating similar organic matter sources throughout the Fruitland coal. These results are similar to 13C reported by Michael et al. (1993) for the Fruitland coals. The isotopic compositions of the Fort Union coals are also relatively constant, 24.4 to 25.8, except for a 13C-enriched value ( 22.7) measured in the deepest portion of the Formation (Table 1). Overall, the isotopic compositions of the bulk organic matter in both basins indicate a consistent input of similar organic matter within each specic basin and our samples.

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that suggests a much lower maturity than deeper in the core. These trends and values are similar to those observed by Michael et al. (1993) for Fruitland coals. For an unknown reason, the C33 hopane ratio in the shallowest Fruitland coal has surpassed typical equilibrium values. The thermal maturity within the Fruitland coal is lower in the shallow coals and increases to equilibrium values with depth. The C31C34 hopane ratios in the Fort Union coal are inconsistent, possibly the consequence of the highly variable, low-grade ranking of the coals. Having previously calculated consistent Ts/(Ts + Tm) ratios that represent lower thermal maturities it is not apparent why these maturity indicators should be so variable. The C31 ratios are low in four samples, ranging from 0.05 to 0.08; however, two anomalous values at depths of 49 and 266 m were observed. Similar observations were made for the other indices; they are generally variable, greatly exceeding equilibrium values or are extremely low, ranging between 0.010.02. If large scale, regional processes raised the thermal maturation in portions of the Fort Union Formation it seems logical that those same processes would have uniformly altered all the Fort Union coals. These enigmatic values may be the result of localized heterogeneities in the organic-rich facies or secondary biodegradation inuences. Even with these inconsistencies, it appears the Fort Union coals are relatively thermally immature, particularly with respect to the Fruitland coals. Overall, the consistent source inputs and thermal maturities measured in these coals provide a foundation to assess the level of biodegradation of the saturated and aromatic hydrocarbons and to address whether this biodegradation is related to the active microbial methanogenesis. 4.4. Effects of biodegradation on saturated hydrocarbons Conventional hierarchies of petroleum or crude oil biodegradation begin with the loss of straight chain alkanes, followed sequentially by acyclic isoprenoids, highly-branched and cyclic saturated hydrocarbons (Peters and Moldowan, 1993; Wegner et al., 2001; Head et al., 2003; Peters et al., 2005). The loss of these GC-resolvable components contributes to an increase in the unresolved complex mixture (UCM), a common qualitative characteristic of biodegraded bitumen (Connan, 1984; Peters and Moldowan, 1993; Peters et al., 2005; Rabus, 2005). Overall, biodegradation in petroleum or crude oil generally proceeds with the removal of saturated hydrocarbons followed by the loss of aromatic hydrocarbons. Fort Union coals show elevated biodegradation indices at several depths (49, 146, 224 m) for the short-, medium-, and long-chain nalkanes and acyclic isoprenoids, typical of conventional biodegraded petroleum. In spite of much higher values for maturity indicators, the Fruitland coals follow a pattern similar to the Fort Union coals for the removal of n-alkanes and acyclic isoprenoids at particular depths (132 and 760 m). The series of nC18nC27-alkylcyclohexanes in the Fruitland coals may be a product of the thermal alteration of the Fruitland coals, as alkylcyclohexanes can be generated during thermal alteration (Kenig, 2000; Brocks and Summons, 2003). The biodegradation indices for these compounds (Table 3) also show preferential removal in the shallowest coals. The biodegradation of these n-alkylcyclohexanes is additional evidence that biodegradation can develop and proceed in more thermally-mature coals in a manner similar to thermally immature rocks. The deepest coal in the Fruitland (1190 m) has equilibrated maturity ratios and the lowest biodegradation indices, suggesting that this is the nonbiodegraded parent coal material. A similar unaltered coal is not found in the Fort Union, implying that coals even in the deepest settings have experienced some level of biodegradation. Overall, the biodegradation in the Fort Union and Fruitland coals has resulted in the removal of n-alkanes and acyclic isoprenoids, which would correspond to a Level of Biodegradation equal to 23 on the scale of Peters and Moldowan (1993), or a level of Slight to

Moderate on the scale of Wegner et al. (2001). The generation and subsequent biodegradation of the alkylcyclohexanes in the Fruitland coal suggest an even higher level of biodegradation than what is indicated by the removal of n-alkanes and acyclic isoprenoids. According to the biodegradation hierarchy of Peters and Moldowan (1993) and Wegner et al. (2001) the removal of alkylcyclohexanes does not occur until Level 34, or Moderate to Heavy. The highest degrees of biodegradation are conned to particular coal seams (Table 3, Fig. 6), suggesting that zones of biodegradation may ultimately be controlled by stratigraphic variation in the subsurface. These heterogeneities in the stratigraphy, including fractures and cleat systems in coals, allowing localized groundwater ow thru particular strata, may stimulate the microbial biodegradation of hydrocarbons. 4.5. Effects of biodegradation on aromatic hydrocarbons In conventional biodegraded petroleum, the biodegradation of aromatic hydrocarbons, such as naphthalenes and phenanthrenes, only occurs after there has been major alteration, and sometimes complete removal, of the n-alkanes (Peters and Moldowan, 1993; Wegner et al., 2001; Head et al., 2003; Peters et al., 2005). However, studies by Ahmed et al. (1999) and Ahmed and Smith (2001) proposed that the biodegradation of aromatic compounds in coals occurs prior to and at a faster rate than the removal of the n-alkanes. For the Fruitland and Fort Union coals only the trimethyl- and tetramethylnaphthalenes (TrMN and TeMN, respectively) could provide useful biodegradation indices because the other substituted naphthalenes were not consistently present throughout the coals. Though, when present, dimethylnaphthalene (DMN) biodegradation indices are similar to TrMN and TeMN. It is likely the naphthalene, mono- and dimethylnaphthalenes were at one point present, but have subsequently been removed by biodegradation in the Fort Union coals. The Fruitland coals indicate extensive removal of naphthalenes in the shallowest depths, and a general decrease in the aromatic biodegradation indices with depth (Table 4, Fig. 7). The Fort Union also has elevated biodegradation indices for the naphthalenes, though the depth relationships are not as apparent as those observed in the Fruitland (Table 4, Fig. 8). The trimethylnaphthalenes appear more biodegraded than the tetramethylnaphthalenes in both coals, which is in accordance with the increasing resistance to biodegradation with increasing alkylation. The biodegradation indices in the phenanthrene series are almost identical to the naphthalene series. Both the Fort Union and Fruitland coals exhibit biodegradation of all the alkyl-substituted phenanthrenes, except for the tetramethylphenanthrenes in the Fort Union coal which could not be calculated due to the low abundances (Table 4, Figs. 7 and 8). In both systems, the same depths exhibit the highest degrees of biodegradation among saturated and aromatic compounds. Unlike the naphthalene series there is no apparent increasing resistance to biodegradation that corresponds to the level of substitution for the phenanthrene series for the Fruitland coal. The Fort Union coals do show progressively less biodegradation with increasing numbers of methyl side groups. According to the aromatic biodegradation hierarchy established by Fisher et al. (1998), the Fort Union and Fruitland coals, which show the removal and alteration of all the alkyl-substituted naphthalenes and phenanthrenes, would suggest a relatively major level of alteration, ranging from moderate to extensive. The scale of Peters and Moldowan (1993) would suggest similarly moderate to heavy levels of biodegradation and the extent of biodegradation would be moderate, level 4 or 5, according to Volkman et al. (1984). These classication schemes of biodegradation hierarchy are based upon petroleum and crude oil degradation and when utilized to assess biodegradation of coal may inaccurately represent the actual level of biodegradation. Aromatic hydrocarbon biodegradation in petroleum or crude oils indicates extensive biodegradation, resulting in a classication of moderate to heavy levels, in coals the biodegradation

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of aromatic hydrocarbons may not be evidence of such extensive biodegradation but may be indicative of moderate to low levels of biodegradation. Based upon our results and the results of Ahmed et al. (1999) and Ahmed and Smith (2001), it may be possible that comparing coal and sedimentary organic matter biodegradation to petroleum and crude oils is insufcient and it may be necessary to develop a specic biodegradation scheme for coals and sedimentary hosted organic matter. Though these biodegradation indices suggest the occurrence of biodegradation in these coals, the inuence or effects of water washing cannot be completely excluded. According to Volkman et al. (1984), benzene, toluene, and C2- and C3-alkyl-substituted benzenes are very soluble in water, and napththalene, methylnaphthalene, and phenanthrene are less soluble. In addition, C2-napthalanes or higher molecular weight aromatic hydrocarbons, e.g. trimethylphenanthrene, are even less water soluble and therefore more resistant to water washing. The presence of an extensive suite of alkyl-substituted naphthalenes and phenanthrenes in both the Fort Union and Fruitland coals would suggest minor, if any, water washing has occurred. Relationships between ne-scale stratigraphy and localized groundwater movement may be controlling the observed distribution of biodegraded coals. Biodegradation of both saturated and aromatic hydrocarbon appears focused in particular, perhaps shallower, coal seams. The biodegradation of aromatic and saturated hydrocarbons, localized in particular depths, suggest that ne-scale groundwater movement may ultimately control the stimulation of microbial methanogenesis. While these observations do not exclude the potential for water-washing of the coals, pristane/nC17 and phytane/ nC18 ratios in both the Fruitland and Fort Union coals suggest an increasing predominance of isoprenoids as other biodegradation indices increase (Table 3) which is more likely the result of biodegradation than water-washing. The occurrence of the more polar aromatic compounds in both coals suggests limited, if any, removal of compounds by water-washing. Also, the presence of thermogenically produced gas in the San Juan Basin does not seem to inuence the biodegradation indices in the shallow Fruitland coals, but the thermal imprints may have masked biodegradation in the deep coals. Overall, it appears these differences in biodegradation indices reect hydrocarbon decomposition directly associated with methanogenesis similar to other eld and laboratory based observations (Ahmed et al. 1999; Zengler et al., 1999; Anderson and Lovely, 2000; Ahmed and Smith, 2001) in two of the world's largest coalbed methane reservoirs. The biodegradation signatures preserved in the coals do not specically follow the observed biodegradation trends in petroleum, crude oil, or in shale-hosted gas reservoirs, but instead show elevated aromatic compound biodegradation prior to complete or extensive removal of more aliphatic compounds in both thermally mature and thermally immature coal seams. 5. Conclusions Microbial degradation of organic matter in the subsurface is well documented in marine sediments and hydrocarbon reservoirs. However, minimal research been focused on linking subsurface coal biodegradation to active microbial methanogenesis. Traditionally, gas compositions and formation water geochemistry have been used to determine the presence of biogenic gas, but the microbial utilization of sedimentary organic matter involved with these processes remains poorly understood. Results of this study reveal the following: (1) biodegradation has occurred at depths up to several hundred meters in both the Fort Union coal of the Powder River Basin and the Fruitland coal of the San Juan Basin, (2) similar to petroleum reservoirs, signatures of biodegradation in the Fort Union and Fruitland coals include extensive removal of n-alkanes and isoprenoids, (3) in contrast to petroleum and crude oil, polycyclic aromatic hydrocarbons in the Fort Union and Fruitland coals are altered prior to the extensive,

if not complete, removal of n-alkanes and acyclic isoprenoids, (4) biodegradation of aliphatic and aromatic hydrocarbons can occur, and in a similar fashion, across a range of inherited thermal maturities, and (5) biodegradation in coalbed methane reservoirs produces similar and recognizable biodegradation signatures that differ from biodegraded petroleum, crude oil, and sedimentary organic matter. Overall, these preliminary results suggest that biodegradation in both thermally immature and mature coals associated with coalbed methane results in similar molecular signatures. However, in order to fully understand the mechanisms responsible for the biodegradation of coal and the biomarker signatures that are indicative of these processes more research is required. Specically, more research is necessary to link the signatures of organic matter biodegradation to active microbial methanogenesis. Acknowledgements We thank Jeanine Honey at the USGS Core Repository in Denver, Colorado, for her assistance in acquiring samples. We also would like to thank James H. Rufn at National Petrographic Service Inc. for the vitrinite reectance data. Additional thanks goes to Elizabeth Gordon of the University of Massachusetts Amherst Biogeochemistry and Stable Isotope Laboratories for analyzing the bulk 13C values of the coals. Helpful comments were provided by Dr. Romeo Flores and two anonymous reviewers. This research was funded by grants from the National Science Foundation Biogeosciences Program to STP (EAR0433766) and AMM (EAR-0433801). References
Ahmed, M., Smith, J.W., 2001. Biogenic methane generation in the degradation of eastern Australian Permian coals. Organic Geochemistry 32, 809816. Ahmed, M., Smith, J.W., George, S.C., 1999. Effects of biodegradation on Australian Permian coals. Organic Geochemistry 30, 13111322. Anderson, R.T., Lovely, D.R., 2000. Hexadecane decay by methanogenesis. Nature 404, 722723. Ayers Jr., W.B., 1986. Coal occurrences of the Tongue River Member, Fort Union Formation, Powder River Basin, Wyoming. Geological Survey of Wyoming Report of Investigations 35, 121. Ayers Jr., W.B., 2002. Coalbed gas systems, resources and production and a review of contrasting cases from the San Juan and Powder River Basins. American Association of Petroleum Geologists Bulletin 86, 18531890. Ayers Jr., W.B., Ambrose, W.A., 1990. Geologic controls on the occurrence of coalbed methane, Fruitland Formation, San Juan basin. In: Ayers, W.B., Kaiser, W.R., Swartz, T.E., Laubach, S.E., Tremain, C.N., WhiteheadIII III, N.H. (Eds.), Geologic Evaluation of Critical Production Parameters for Coalbed Methane Resources, Part 1: San Juan Basin: Chicago, Gas Research Institute Annual Report, GRI 90/0014.1, pp. 972. Ayers Jr., W.B., Kaiser, W.R., 1994. Lacustrine interdeltaic coal in the Fort Union Formation (Paleocene) Powder River basin, Wyoming and Montana, U.S.A. In: Rahmani, R.A., Flores, R.M. (Eds.), Sedimentology of Coal and Coal-Bearing Sequences. International Association of Sedimentologists Special Publication, vol. 7, pp. 6184. Ayers Jr., W.B., Kaiser, W.R., Laubach, S.E., Ambrose, W.A., Baumgardner Jr., R.W., Scott, A.R., Tyler, R., Yeh, J., Hawkins, G.J., Swartz, T.E., Schultz-Ela, D.D., Zellers, S.D., 1991. Geologic and Hydrologic Controls on the Occurrence and Producibility of Coalbed Methane, Fruitland Formation. Gas Research Institute Topical Report, GRI 91/0072. 314 pp. Bishop, A.N., Abbot, G.D., 1993. The interrelationship of biological marker maturity parameters and molecular yields during contact metamorphism. Geochimica Cosmochimica Acta 57, 36613668. Bray, E.E., Evans, E.D., 1961. Distribution of n-parafns as a clue to recognition of sources beds. Geochimica,et Cosmochimica Acta 22, 215. Brocks, J.J., Summons, R.E., 2003. Sedimentary hydrocarbons, biomarkers for early life. In: Schlesinger, W.H. (Ed.), Treatise on Geochemistry, vol. 8. Elsevier, Oxford, pp. 63115. Brocks, J.J., Love, G.D., Summons, R.E., Knoll, A.H., Logan, G.A., 2005. Biomarker evidence for green and purple sulphur bacteria in a stratied Palaeoproterozoic sea. Nature 437, 866870. Brister, B.S., Hoffman, G.K., 2002. Fundamental geology of San Juan Basin energy resources. In: Brister, B.S., Greer Price, L. (Eds.), New Mexico's Energy, Present and Future. Decision-Makers Field Conference 2002, pp. 2138. Cannico, U., Tocco, R., Ruggiero, A., Surez, H., 2004. Organic geochemistry and petrology of coals and carbonaceous shales from western Venezuela. International Journal of Coal Geology 57, 151165. Clayton, J.L., Rice, D.D., Michael, G.E., 1991. Oil-generating coals of the San Juan Basin, New Mexico and Colorado, U.S.A. Organic Geochemistry 17, 732745.

M. Formolo et al. / International Journal of Coal Geology 76 (2008) 8697 Connan, J., 1984. Biodegradation of crude oils in reservoirs. In: Brooks, J., Welte, D.H. (Eds.), Advances in Petroleum Geochemistry. Academic Press, London, pp. 299335. Craigg, S.D., 2001. Geologic framework of the San Juan structural basin of New Mexico, Colorado, Arizona, and Utah, with emphasis on Triassic through Tertiary rocks. U.S. Geological Survey Professional Paper 1420 70 pp.. Curry, D.J., Emmett, J.K., Hunt, J.W., 1994. Geochemistry of aliphatic-rich coals in the Cooper Basin, Australia and Taranaki Basin, New Zealand: implications for the occurrence of potentially oil-generative coals. In: Scott, A.C., Fleet, A.J. (Eds.), Coal and Coal-Bearing Strata as Oil-Prone Source Rocks. Geological Society Special Publication, London, vol. 77, pp. 149182. Eglington, G., Hamilton, R.J., 1967. Leaf epicuticular waxes. Science 156, 13221335. Ensminger, A., van Dorsselaer, A., Spyckerelle, C., Albrecht, P., Ourisson, G., 1974. Pentacyclic triterpenes of the hopane type as ubiquitous geochemical markers: origin and signicance. In: Tissot, B., Bienner, F. (Eds.), Advances in Organic Geochemistry. Editions Technip. Paris, pp. 245260. Farrimond, P., Taylor, A., Talns, N., 1998. Biomarker maturity parameters: the role of generation and thermal degradation. Organic Geochemistry 29, 11811197. Fassett, J.E., 1988. Geometry and depositional environment of Fruitland Formation coal beds, San Juan basin, New Mexico and Colorado: anatomy of a giant coal-bed methane deposit. In: Fassett, J.E. (Ed.), Geology and Coalbed Methane Resources of the Northern San Juan Basin, Colorado and New Mexico. Rocky Mountain Association of Geologists, Denver, pp. 2338. Fassett, J.E., Hinds, J.S., 1971. Geology and Fuel Resources of the Fruitland Formation and Kirtland Shale of the San Juan Basin, New Mexico and Colorado, vol. 676. United States Geological Survey Professional Paper. 76 pp. Fisher, S.J., Alexander, R., Kagi, R.I., 1996. Biodegradation of alkylnaphthalenes in sediments adjacent to an off-shore petroleum production platform. Polycyclic Aromatic Compounds 11, 3542. Fisher, S.J., Alexander, R., Kagi, R.I., Oliver, G.A., 1998. Aromatic hydrocarbons as indicators of biodegradation in North Western Australian reservoirs. In: Purcell, P.G., Purcell, R.R. (Eds.), The Sedimentary Basins of Western Australia 2. Proceedings of Petroleum Exploration Society of Australian Symposium, Perth, WA, pp. 185194. Flores, R.M., 2004. Coalbed Methane in the Powder River Basin, Wyoming and Montana: An Assessment of the Tertiary-Upper Cretaceous Coalbed Methane Total Petroleum System. U.S. Geological Survey Digital Data Series DDS-69-C. 56 pp. Flores, R.M., Etheridge, F.G., 1985. Evolution of intermontane uvial systems of Tertiary Powder River basin, Montana and Wyoming. In: Flores, R.M., Kaplan, S.S. (Eds.), Cenozoic Paleogeography of WestCentral United States. Society of Economic Paleontologists and Mineralogists Rock Mountain Section, Rocky Mountain Paleogeography Symposium, vol. 3, pp. 107126. Flores, R.M., Rice, C.A., Stricker, G.D., Warden, A., Ellis, M.S., 2008. Methanogenic pathways of coalbed gas in the Powder River Basin, United States: the geologic factor. International Journal of Coal Geology 76, 5275 (this volume). doi:10.1016/j. coal.2008.02.005. Formolo, M.J., Salacup, J.M., Petsch, S.T., Martini, A.M., Nsslein, K., 2008. A new model linking atmospheric methane sources to Pleistocene glaciation via methanogenesis in sedimentary basin. Geology 36, 139142. Gorody, A.W., 1999. The origin of natural gas in the Tertiary coal seams on the eastern margin of the Powder River basin. Wyoming Geological Association Guidebook 50, 89101. Head, I.M., Jones, D.M., Larter, S.R., 2003. Biological activity in the deep subsurface and the origin of heavy oil. Nature 426, 344352. Kaiser, W.R., Swartz, T.E., Hawkins, G.J., 1991. Hydrology of the Fruitland Formation, San Juan Basin. In: AyersJr. Jr., W.B., et al. (Ed.), Geologic and Hydrologic Controls on the Occurrence and Producibility of Coalbed Methane, Fruitland Formation, San Juan Basin. Bureau of Economic Geology. University of Texas Report, Contract No. 50872124-1544, pp. 159241. Kenig, F., 2000. C16C29 homologous series of monomehthylalkanes in the pryrolysis products of a Holocene microbial mat. Organic Geochemistry 31, 237241. Law, B.E., 1992. Thermal maturity patterns of Cretaceous and Tertiary rocks, San Juan basin, New Mexico and Colorado. U.S. Geological Survey Open-File Report 90-659, p. 18. Law, B.E., Rice, D.D., Flores, R.M., 1991. Coalbed gas accumulations in the Paleocene Fort Union Formation, Powder River basin, Wyoming. In: Schwochow, S.D., Murray, D.K., Fahy, M.F. (Eds.), Coalbed Methane of Western North America. Rocky Mountain Association of Geologists, pp. 179190. Lucas, S.P., Hunt, A.P., Sullivan, R.M., 2006. Stratigraphy and age of the Upper Cretaceous Fruitland Formation, west-central San Juan Basin, New Mexico. In: Lucas, S.G., Sullivan, R.M. (Eds.), Later Cretaceous Vertebrates From the Western Interior. New Mexico Museum of Natural History and Science Bulletin, vol. 35, pp. 16. Martini, A.M., Budai, J.M., Walter, L.M., Schoell, M., 1996. Microbial generation of economic accumulations of methane within a shallow organic-rich shale. Nature 383, 155158. Martini, A.M., Walter, L.M., Budai, J.M., Ku, T.C.W., Kaiser, C.J., Schoell, M., 1998. Genetic and temporal relations between formation waters and biogenic methane. Geochimica,et Cosmochimica Acta 62, 16991720.

97

McIntosh, J.C., Walter, L.M., Martini, A.M., 2004. Extensive microbial modication of formation water geochemistry: case study from a midcontinent sedimentary basin, United States. Geological Society of America Bulletin 116, 743759. Michael, G.E., Anders, D.E., Law, B.E., 1993. Geochemical evaluation of Upper Cretaceous Fruitland Formation coals, San Juan Basin, New Mexico and Colorado. Organic Geochemistry 20, 457498. Montgomery, S.L., 1999. Powder River basin, Wyoming: an expanding coalbed methane (CBM) play. American Association of Petroleum Geologists Bulletin 83, 12071222. Nuccio, V.F., 1990. Burial, thermal, and petroleum generation history of the Upper Cretaceous Steele Member of the Cody Shale (Shannon Sandstone bed horizon), Powder River Basin, Wyoming. U.S. Geological Survey Bulletin 1917-A 17 pp. Pashin, J.C., 1998. Stratigraphy and structure of coalbed methane reservoirs in the United States: an overview. International Journal of Coal Geology 35, 209240. Peters, K.E., Moldowan, J.M., 1993. The Biomarker Guide: Interpreting Molecular Fossils in Petroleum and Ancient Sediments. Prentice-Hall, New Jersey. 363 pp. Peters, K.E., Walters, C.C., Moldowan, J.M., 2005. The Biomarker Guide: Volume II Biomarkers and Isotopes in Petroleum Exploration and Earth History. Cambridge University Press, United Kingdom. 680 pp. Rabus, R., 2005. Biodegradation of hydrocarbons under anoxic conditions. In: Ollivier, B., Magot, M. (Eds.), Petroleum Microbiology. American Society for Microbiology ASM -Press, Washington D.C., pp. 277299. Rice, D.D., 1983. Relation of natural gas composition to thermal maturity and source rock type in San Juan Basin, northwestern New Mexico and southwestern Colorado. American Association of Petroleum Geologists Bulletin 67, 11191218. Rice, D.D., 1993. Composition and origins of coalbed gas. Hydrocarbons from coal. American Association of Petroleum Geologists Studies in Geology 38, 159184. Rice, D.D., Clayton, J.L., Pawlewicz, M.J., 1989. Characterization of coal-derived hydrocarbons and source-rock potential of coal beds, San Juan Basin, New Mexico and Colorado, U.S.A. International Journal of Coal Geology 13, 597626. Robinson Roberts, L.N., McCabe, P.J., 1992. Peat accumulation in coastal-plain mires: a model for coals of the Fruitland Formation (Upper Cretaceous) of southern Colorado, U.S.A. International Journal of Coal Geology 21, 115138. Scalan, R.S., Smith, J.E., 1970. An improved measure of the oddeven predominance in the normal alkanes of sediment extracts and petroleum. Geochimica et Cosmochimica Acta 34, 611620. Scott, A.R., Kaiser, W.R., Ayers Jr, W.B., 1991. Composition, distribution, and origin of Fruitland Formation and Pictured Cliffs Sandstone gases, San Juan Basin, Colorado and New Mexico. In: Schwochow, S.D., Murray, D.K., Fahy, M.F. (Eds.), Coalbed Methane of Western North America. Rocky Mountain Association of Geologists Field Conference Guidebook, pp. 93108. Scott, A.R., Kaiser, W.R., Ayers Jr., W.B., 1994. Thermogenic and secondary biogenic gases, San Juan basin, Colorado and New Mexico implications for coalbed gas producibility. American Association of Petroleum Geologists Bulletin 76, 11861209. Siefert, W.K., Moldowan, J.M., 1978. Application of steranes, terpanes, and monoaromatics to the maturation, migration and source of crude oils. Geochimica et Cosmochimica Acta 42, 7795. Seifert, W.K., Moldowan, J.M., 1986. Use of biological markers in petroleum exploration. In: Jones, R.B. (Ed.), Biological Markers in the Sedimentary Record, Methods in Geochemistry and Geophysics. Elsevier Publishing, Amsterdam, pp. 261290. Stricker, G.D., Flores, R.M., Trippi, M.H., Ellis, M.S., Olson, C.M., Sullivan, J.E., Takahashi, K. I., 2007. Coal quality and major, minor, and trace elements in the Powder River, Green River, and Williston Basins, Wyoming and North Dakota. U.S. Geological Survey Open-File Report 20071117. 31 pp. Skorov, I., Pickel, W., Christanis, K., Wolf, M., Taylor, G.H., Flores, D., 2005. Classication of huminite ICCP System 1994. International Journal of Coal Geology 62, 85106. Tissot, B.P., Welte, D.H., 1984. Petroleum Formation and Occurrence, 2nd edition. Springer-Verlag, Berlin. 699 pp. Tyler, R., Kaiser, W.R., Scott, A.R., Hamilton, D.S., Ambrose, W.A., 1995. Geologic and hydrologic assessment of natural gas from coal: Greater Green River, Piceance, Powder River, and Raton basins, western United States. University of Texas at Austin, Bureau of Economic Geology Report of Investigations, vol. 222. 219 pp. Volkman, J.K., Alexander, R., Kagi, R.I., Rowland, S.J., Sheppard, P.N., 1984. Biodegradation of aromatic hydrocarbons in crude oils from the Barrow Sub-basin of Western Australia. Organic Geochemistry 6, 619632. Waples, D.W., Machihara, T., 1991. Biomarkers for Geologists: a practical guide to the application of steranes and triterpanes in petroleum geology. American Association of Petroleum Geologists Methods in Exploration Series No. 9. Tulsa. 91 pp. Wegner, L.M., Davis, C.L., Isaksen, G.H., 2001. Multiple controls on petroleum biodegradation and impact on oil quality. Society of Petroleum Engineers Reservoir Evaluation and Engineering 5, 375383. Zengler, K., Richnow, H.H., Rosello-Mora, R., Michaelis, W., Widdel, F., 1999. Methane formation from long-chain alkanes by anaerobic microorganisms. Nature 401, 266269. Zhou, Z., Ballentine, C.J., Kipfer, R., Schoell, M., Thibodeaux, S., 2005. Noble gas tracing of groundwater/coalbed methane interaction in the San Juan Basin, USA. Geochimica, et Cosmochimica Acta 69, 54135428.