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Methane production from horse manure and

stall waste with softwood bedding

Article in Bioresource Technology · February 2012

DOI: 10.1016/j.biortech.2012.02.012 · Source: PubMed

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Contents lists available at SciVerse ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Methane production from horse manure and stall waste with softwood bedding
Brian A. Wartell, Valdis Krumins, Jeffrey Alt, Kathleen Kang, Bryan J. Schwab, Donna E. Fennell ⇑
Department of Environmental Sciences, 14 College Farm Road, Rutgers University, New Brunswick, NJ 08901, USA

a r t i c l e i n f o a b s t r a c t

Article history: Substantial stall waste is generated from horses on softwood bedding. The methane potential (Gpot) of
Received 21 November 2011 horse manure and constructed mixtures of stall waste with softwood bedding was determined at
Received in revised form 31 January 2012 35 °C. Gpot of 68, 191 and 273 mL/g volatile solids (VS) were estimated for three separate batches of horse
Accepted 2 February 2012
manure, indicating variability in the material. Cumulative energy production over 20–40 days ranged
Available online xxxx
from 3.11 ± 0.92 to 8.45 ± 5.42  105 kJ/metric ton wet weight horse manure alone, and from
1.69 ± 0.39 to 3.91 ± 0.47  105 kJ/metric ton wet weight horse manure plus softwood stall bedding
Keywords:
(mixed at a 1:1 ratio on a VS basis). Softwood bedding was barely degradable and diluted the energy pro-
Stall waste
Horse manure
duction of the stall waste; however, it did not cause inhibition of methane production from manure. Man-
Softwood bedding ually separated used softwood bedding contained substantial methane potential.
Anaerobic digestion Ó 2012 Elsevier Ltd. All rights reserved.
Methane potential

1. Introduction softwood, which contains compounds with known toxic properties

The equine industry faces a substantial waste disposal problem
whose cost could be partially offset by energy recovery through
anaerobic digestion. One horse (a 454 kg animal) produces roughly
17 kg feces and 9 L of urine per day, for a total of 27 kg of waste
(Romano et al., 2006; Westendorf and Krogmann, 2004; Wheeler
and Zajaczkowski, 2002). Stalled horses require up to 9 kg of bed-
ding per day (Westendorf and Krogmann, 2004; Wheeler and
Zajaczkowski, 2002). Under these circumstances, combined bed-
ding and manure accounts for up to 12 metric tons (12,000 kg) of
wet waste per horse per year.
Most equine waste that could be recovered and considered for
anaerobic digestion consists of manure plus some type of stall bed-
ding (Westendorf and Krogmann, 2006; Wheeler and Zajaczkow-
ski, 2002). The waste characteristics vary substantially depending
upon the type of bedding utilized and the frequency and nature
of stall cleaning, e.g. spot cleaning versus complete removal of bed-
ding and manure (Airaksinen, 2006; Chamberlain et al., 2004;
Westendorf and Krogmann, 2006). Softwood (e.g., pine or fir) shav-
ings or chips are often used as bedding because of high absorbency,
low palatability and low cost (Airaksinen, 2006; Chamberlain et al.,
2004). Straw is preferentially used as bedding for mares with foals
because of its low toxicity (Airaksinen, 2006) when compared with
⇑ Corresponding author. Address: 14 College Farm Road, Department of Environ-
mental Sciences, Room 231, Environmental and Natural Resources Sciences
Building, Rutgers University, New Brunswick, NJ 08901, USA. Tel.: +1 732 932
9800x6204; fax: +1 732 932 8644.
E-mail address: fennell@envsci.rutgers.edu (D.E. Fennell).
(Belmonte et al., 2006; Savluchinske-Feio et al., 2006). There is a
market for stall waste from horses bedded on straw via the mush-
room production industry (Poppe, 2000) and depending upon geo-
graphic location, this may also affect the choice of bedding for a
specific facility.
A 2007 assessment of biomass in New Jersey (NJ) showed that
approximately 102,400 dry tons equine stall waste per year is
recoverable; an amount greater than all other agricultural livestock
waste combined (Brennan et al., 2007). Approximately 75% of
horse farms utilize or store manure on-site and for those farms
not utilizing off-site hauling, the application of manure and stall
waste on fields and pastures is the primary means of disposal
(Warren, 2003; Westendorf et al., 2010). Equine waste produces
odors and could contaminate water sources via runoff during stor-
age or after land application (Airaksinen et al., 2006; McFarland,
2008; Romano et al., 2006). Because many facilities need to store
or haul stall waste (Westendorf et al., 2010), an increasing cost is
borne by the farmer to handle this material.
Only a few groups have investigated anaerobic digestion of
horse waste (Kalia and Singh, 1998; Mandal and Mandal, 1998;
Zuru et al., 2004; Kusch et al., 2008; Cui et al., 2011). This may
be because it is a less abundant waste overall than cattle and swine
manures and because the higher solids content of horse waste
makes this material highly suitable for composting (Chamberlain
et al., 2004; Krogmann et al., 2006; Westendorf and Krogmann,
2004; Wheeler and Zajaczkowski, 2002).
We previously investigated semi-continuous laboratory-scale
anaerobic digestion of horse manure and observed a substantial
drop in methane production after the feedstock was changed from
manure to softwood chip-based stall waste (Wartell et al., 2008).

0960-8524/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2012.02.012

Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
2 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx
To our knowledge there is no other information available on the
methane production potential of stall waste with softwood-based
bedding. Here we report the methane production potential of horse
manure, mixtures of horse manure and fresh or used softwood
(pine) chips, softwood pellets or straw, and softwood pellet-based
stall waste from a working equine facility. We further investigate
whether increasing ratios of softwood bedding have an inhibitory
effect on methane production.
2. Methods
2.1. Feed stock and inoculum
2.1.1. Feedstocks for methane potential and toxicity tests
Fresh horse manure without bedding was collected from accu-
mulated waste in loafing sheds or stalls where horses urinated
and defecated on a daily basis. Fresh softwood chips, Woody
PetÒ, a softwood pellet product (Woody Pet, Surrey, BC, Canada)
and straw bedding were provided by the New Jersey Agricultural
Experiment Station (NJAES) Animal Care Program at Rutgers Uni-
versity, New Brunswick, NJ and staff from the Rutgers University
Equine Science Center. Used softwood chip bedding previously ex-
posed to urine and manure was manually separated from actual
stall waste using tweezers. Stall waste with softwood bedding
was collected from stalls at the Animal Care Program at Rutgers
University.
2.1.2. Feedstock for solid state batch test in 125 L reactors
A solid state batch digester study was performed using 125 L
reactors loaded with stall waste obtained from an equine facility
operating in northern NJ. The waste was generated from stalls bed-
ded with Condensed Pine Wood Bedding Pellets (Guardian Horse
Bedding, Equistock, LLC, Rockford, IL). Stalls were spot cleaned
twice per day. Based on the number of horses on site (60) and
the amount of bedding purchased per year, it was estimated that
the waste consisted of 1:1–2:1 softwood bedding volatile solids
(VS) to manure VS. The waste had been stored on site in static piles
under winter conditions for approximately two weeks prior to use.
Elevated temperatures in the piles suggested some aerobic degra-
dation was occurring.
2.1.3. Inoculum
Sludge from a mesophilic anaerobic digester at the Joint Meet-
ing of Essex and Union Counties Edward P. Decker 32,200 m3/d
Secondary Wastewater Treatment Facility located in Elizabeth, NJ
Table 1
Methane potential batch testsa for horse manure and stall bedding.
Exp. Test
Exp. 1 Effect of fresh softwood bedding
Exp. 2 Effect of fresh softwood bedding
Exp. 3 Effect of bedding Methane potential of bedding alone
Exp. 4 Effect of fresh and used softwood bedding
Exp. 5 Effect of fresh softwood bedding
Methane potential of bedding alone
a
Experimental bottles were inoculated with 10 mL of inoculum, filled with anaerobic minimal medium to 100 mL and operated at 35 °C.
Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
was used as methanogenic inoculum. All materials were stored
at 4 °C prior to use.
2.2. Methane production potential tests
Five separate batch reactor experiments were performed as
shown in Table 1. Horse manure alone, horse manure mixed with
increasing ratios of different types of stall beddings, and bedding
alone, were tested. The methane production potential of horse
manure alone was examined at bottle loadings of 0.5 and 2.38 g
VS/bottle and spanning 33–79 days (Table 1). The beddings tested
were softwood bedding (fresh and used (viz., bedding manually
separated from stall waste as described in Section 2.1.1)), Woody
PetÒ, a softwood pellet bedding that disintegrates into small wood
particles under the influence of moisture, and straw. Each bedding
type was also tested alone (without manure) to determine the rel-
ative contribution of bedding degradation to total methane
production.
Note that based on the TS and VS of manure and bedding listed
in Table 2 and assuming a typical amount of bedding (9 kg) and
manure (17 kg) per horse per day (Romano et al., 2006; Westen-
dorf and Krogmann, 2004; Wheeler and Zajaczkowski, 2002), a
typical bedding VS to manure VS ratio would be approximately
1.4:1 for typical uses of softwood chip bedding. We investigated
methane production from mixtures containing bedding VS to horse
manure VS ratios of 0.01:1, 0.05:1, 0.10:1, 0.25:1, 0.5:1, 1:1, 2:1
and 4:1. The increasing ratios of bedding to manure were used to
evaluate the impact of bedding on the energy content of the mate-
rial and also to determine whether there was any inhibitory effect
on methanogenesis by the presence of the bedding. The possibility
that exposure to urine and the elements might enhance or dimin-
ish any inhibitory effect of the softwood bedding was also tested
using admixtures of horse manure with the manually separated,
used softwood bedding, in addition to the tests with the fresh, un-
used softwood chip bedding (Table 1).
Methane production was determined via batch tests in 160 mL
serum bottles established under anaerobic conditions. Waste
materials were added to serum bottles under an oxygen-free nitro-
gen purge. Municipal anaerobic digester sludge (10 mL) was added
as inoculum, then anaerobic minimal medium (Fennell et al., 1997)
was added to bring the total operating volume to 100 mL. The reac-
tors were vigorously shaken by hand, and then incubated at 35 °C
with occasional manual shaking, for the durations indicated in Ta-
ble 1. Each experiment included inoculum-only control treatments
to determine the methane production from decay of the inoculum.
Substrates Duration Loadings
(d)
Manure (g VS) Stall bedding ratio (g stall bedding
VS: g manure VS)
Manure 59 0.5 0.01, 0.05, 0.10, or 0.25
Fresh softwood chip bedding
Manure 40 0.5 0.05, 0.10, 0.25 or 0.5
Fresh softwood chip bedding
Manure 46 0.5 0.25, 0.5, 1 or 2
Fresh softwood chip bedding
Fresh pelleted softwood bedding
Fresh straw
Manure 79 2.38 0.25, 0.5, 1, 2 or 4
Fresh softwood chip bedding
Used softwood chip bedding
Manure 33 2.38 0.25, 0.5, 1 or 2
Fresh softwood chip bedding
 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx 3

Table 2
Average total and volatile solids content of the feedstocks (ranges are shown in parentheses).
a
Material Total solids (% wet weight) Volatile solids (% TS)
b
Horse Manure 37.0 (20–42) 83.7 (76–92)
n = 21
c
Stall Waste 32.0 (22–40) 79.8 (79–91)
(manure plus softwood chip bedding)
n = 18
d
Stall waste from active NJ equine facility using Guardian Horse Bedding 41.3 (38.5–42.9) 82.0 (77.9–85.3)
n=3
Softwood Bedding 92.1 (91–93) 90.1 (89–99)
(fresh)
n=6
Softwood Bedding 31.2 (30–32) 92.8 (91–94)
(used, manually separated)
n=6
Woody PetÒ 93.8 (93–94) 90.8 (90–92)
Softwood pellets
n=3
Guardian Horse Bedding 92.2 (91.7–93) 84.5 (83–85.3)
Softwood pellets
n=3
Straw 93.3 (92–94) 97.9 (97–98)
n=3
a
n = total number of replicates analyzed.
b
collected from outdoor loafing sheds.
c
collected from stalls.
d
collected from outdoor piles after 2 weeks storage.

Pressure and data were synced to presentation from Day 0. On Days 50 (Di-
Gauge gester #1) and 43 (Digester #2), respectively, each reactor was
Rubber
Biogas
Gasket
Temp. Probe opened and 10 L additional inoculum (10% volume:volume amend-
Line Connected to ment) was added.
Controller for
Heated Water The biogas flow from each reactor was continually measured by
a wet test meter (Precision Scientific, Chicago, IL). Leachate that
Outlet for
Wet drained from the waste pile to the bottom of the reactor was
Heated Water
Test
Meter pumped to the top of the reactor every 2–5 days. The reactor tem-
perature was maintained by heated tap water supplied by a 75 L
Double Wall electric water heater (Reliance, Ashland City, TN) and recirculated
with Heated
Water through the water jacket of the reactor via a UP15–42 F pump
Stall Waste
(Grundfos, Olathe, KS). Pump operation was controlled by a TA-3
Insulation
controller (SUPCO, Allenwood, NJ) that monitored temperature
Leachate probes inside the reactor and the water jacket. The reactors were
Height : 100 cm
Diameter: 40 cm
Recirculation
Volume: 125 L Line maintained between 34.0 and 36.0 °C (with exception of a distur-
bance in Digester #2 as described in Section 3.4).
Inlet for
Heated
Water
Vessel with 2.4. Analyses
Caster Leachate
Recirculation
Screen (5 mm) Pump 2.4.1. Solids analysis
Leachate Total and volatile solids analyses were performed according to
Sampling standard methods (Clesceri et al., 1998). Total solids (TS) and vol-
atile solids (VS) content of the materials are shown in Table 2. Sam-
Fig. 1. Schematic of 125 L batch reactors for high-solids anaerobic digestion
(Krogmann et al., 2003). ples were tested in at least triplicate.

2.3. Solid state batch tests in 125 L reactors
Anaerobic digestion of stall waste from an operational equine
facility in northern NJ was performed in high solids, batch stain-
less-steel water-jacketed reactors covered with foam-insulation
(Fig. 1, Krogmann et al., 2003). The reactors were equipped with
stainless-steel screens near the bottom so that a waste pile could
be held in place while leachate could be collected at the bottom
of the reactor, then recirculated to the top of the pile. Prior to
use, reactors were tested for ability to hold pressure at approxi-
mately 14 kPa. At start-up, each reactor (denoted Digester #1
and Digester #2) was filled to approximately 100 L with 29 kg
wet weight (9.8 kg VS) stall waste plus 2 L of inoculum (2% vol-
ume:volume amendment). Reactors were initiated 7 days apart,
2.4.2. Biogas and methane measurements
Biogas was released from the 160 mL batch serum bottle reac-
tors every 3 to 4 days and the volume was measured at atmo-
spheric pressure using a gas-tight plastic syringe or a water
displacement system constructed from a 100 or 500 mL burette.
For the solid state 125 L batch reactors, biogas production was
determined by noting the reading on the wet test meter every 1
to 4 days and computing an average daily biogas production rate.
Every 2 to 3 days, an evacuated 3 L TedlarÒ gas bag (CEL Scientific,
Santa Fe Springs, CA) was connected to the outlet of the wet test
meter to obtain a biogas sample from which to determine the
methane content.
Methane content of the biogas was determined using 0.5 mL
biogas samples taken from the headspace of each serum bottle

Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
4 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx

after biogas volume measurement/venting, when the bottle methane (1:1 ratio relative to the manure only control) if no inhi-
headspace was at atmospheric pressure, or from the TedlarÒ gas bition or toxicity occurred.
bag for the 125 L digesters. The gas sample was removed using a
Series A-2 glass-TeflonÒ-stainless-steel gas-tight syringe equipped
3. Results and discussion
with a side port needle (ValcoÒ Precision Sampling, Baton Rouge,
LA) and injected into an AgilentÒ 6890 N gas chromatograph (Agi-
3.1. Methane production during batch tests
lent Technologies, Santa Clara, CA) equipped with a GS-GasPro cap-
illary column (30 m  0.32 mm I.D.; J&W Scientific, Folsom, CA)
The cumulative methane production from horse manure alone
and a flame ionization detector. Helium was the carrier gas at a
was determined in five separate batch tests over incubation times
constant pressure of 131 kPa. The oven temperature was 150 °C.
ranging from 33 to 79 days (Table 1). The total cumulative meth-
The resulting chromatographic peak area was compared to a five-
ane production for horse manure alone was variable depending
point calibration curve prepared using mixtures of 0–99% methane
upon the duration of the run and also the specific batch of manure.
created by mixing volumes of methane (99% purity; Matheson Tri-
Methane production of 56 ± 14, 122 ± 78, 53 ± 15, 231 ± 18 and
Gas, Inc., Montgomeryville, PA) and air in a 0.5 mL gas-tight syr-
133 ± 6 mL/g VS was observed for batch runs over 33, 40, 46, 59
inge (ValcoÒ Precision Sampling, Baton Rouge, LA). Other compo-
and 79 days, respectively. The upper observed values were compa-
nents of the biogas were not analyzed.
rable to our earlier observation from semi-continuous complete

2.5. Data analyses

160
Horse Manure
Used Softwood Bedding-Horse Manure (0.25:1)

Cummulative Methane (mL/g VS)

The methane production for each serum bottle was determined 140 Used Softwood Bedding-Horse Manure (0.5:1)
from the biogas volume and methane content and reported at 25 °C Used Softwood Bedding-Horse Manure (1:1)
Fresh Softwood Bedding-Horse Manure (1:1)
and 1 atm. Cumulative methane production from inoculum alone 120 Used Softwood Bedding-Horse Manure (2:1)
accounted for <1 percent of total cumulative methane production Used Softwood Bedding-Horse Manure (4:1)
Used Separated Softwood Bedding
from manure/bedding mixtures, straw bedding alone and used 100 Fresh Softwood Bedding Alone

bedding; and <10 percent from softwood-based bedding alone,

and were not subtracted from the methane production values re- 80

ported for the different treatments.

60
The potential energy production from the waste expressed as kJ
per metric ton wet waste was determined by dividing the total 40
cumulative volume of methane produced by the total wet weight
of waste (manure and/or bedding) added to each bottle or reactor 20
(converted to metric tons). This amount was then converted to mol
of methane per metric ton and multiplied by the energy potential 0
0 10 20 30 40 50 60 70 80
of the methane, 802 kJ/mol methane (Schwarzenbach et al., 2003). Time (days)
The maximum methane production potential, Gpot (mL/g VS
added) was determined by a non-linear curve fit of batch data as Fig. 2. Cumulative methane production from horse manure and mixtures of fresh
described by Kusch et al. (2008) or used softwood chip bedding plus horse manure based on the total amount of
volatile solids (VS) loaded (mL methane/g VS). The ratios of waste loaded (bedding:
manure) were: 0.25:1 (0.595 g softwood chip bedding VS, 2.38 g manure VS), 0.5:1
GðtÞ ¼ a þ b1 expðk1 tÞ þ b2 expðk2 tÞ ð1Þ (1.19 g softwood chip bedding VS, 2.38 g manure VS); 1:1 (2.38 g softwood chip
bedding VS, 2.38 g manure VS); 2:1 (4.76 g softwood chip bedding VS, 2.38 g
Where G(t) is the methane production at time t and a, bi, and ki
manure VS); and 4:1 (9.52 g softwood chip bedding VS, 2.38 g manure VS). [h,
are constants describing conversion of the material to methane. Horse manure only; }, used softwood bedding – horse manure 0.25:1; s, used
Gpot is the methane production potential as t ? 1. Batch test data softwood bedding - horse manure 0.5:1; N, used softwood bedding – horse manure
from horse manure only (for Exp 1, 3, and 4 operated over 59, 46 1:1; j, fresh softwood bedding - horse manure 1:1; 4, used softwood bedding -
and 79 days, respectively, Table 1) were averaged and the average horse manure 2:1; , used softwood bedding – horse manure 4:1; , used softwood
bedding; and d, fresh softwood bedding.].
data were fit with Equation 1 using the solver function in Microsoft
Excel and minimizing the sum of squares error between the model
and the data. Gpot from mixtures of the horse manure with various
beddings were then determined by using the rate parameters (ki) 70
Cumulative Methane Production

from the corresponding manure batch and fitting the degradable Fresh Straw
fractions (bi) to achieve a best fit. Results are presented as an aver- 60
Used Softwood Chips
age ± one standard deviation. For this analysis, the total VS includ-
50
ing horse manure and/or bedding loaded into the bottles was used Fresh Softwood Chips
(mL/g VS)

for the basis of comparison.

40 Fresh Softwood Pellets
To assess whether increasing relative amounts of bedding were
inhibitory to the digestion process, analysis of variance (ANOVA) 30
was conducted using Microsoft ExcelÒ to determine the statistical
significance of differences between measured methane production 20
potential from manure and bedding mixtures, relative to controls
receiving horse manure only. A significance level of 0.05 was used. 10
In this case, methane production was based on the horse manure
VS loading alone. Because the softwood bedding was determined 0
0 20 40 60 80
to be scarcely degradable, it should have contributed only a small
Batch Incubation Time (days)
portion of the total methane production. Thus, bottles receiving the
same amount of horse manure VS but increasing amounts of soft- Fig. 3. Methane production observed from different bedding types. [N, Straw; s,
wood bedding VS should have produced equivalent amounts of used softwood bedding; }, fresh softwood chips; j, fresh softwood pellets.].

Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx
mix reactors with horse manure alone (Wartell et al., 2008) where
methane production was 183 ± 67 mL CH4/g VS, at a solids reten-
tion time of 24 days.
As an example of the batch results, cumulative methane pro-
duction from horse manure alone and mixtures of horse manure
and used bedding during the 79 day batch test (Exp. 4) is shown
in Fig. 2. The fresh softwood bedding was only minimally degrad-
able under anaerobic conditions (Figs. 2 and 3), thus, as the ratio of
softwood-based bedding to horse manure was increased, the
methane production on a total volatile solids loading basis de-
creased (Fig. 2). Horse manure mixed with fresh or used softwood
bedding at a 1:1 bedding VS to horse manure VS ratio produced
half or less of the methane that was produced by horse manure
alone (Fig. 2). The methane potential observed for both softwood
bedding chips and pellets (<10 mL/g VS regardless of incubation
time) (Fig. 3) reflect the low values typically observed for softwood
biomass. For example, Tong et al. (1990) reported a methane po-
tential of 42 mL/g VS for white fir and (Chynoweth et al., 1985) re-
ported a methane potential of 63 mL/g VS for loblolly pine;
however Kim and Townsend (2007) reported a methane potential
<20 mL/g VS, a value indistinguishable from inoculum alone in that
study, for yellow pine. Used softwood bedding separated from stall
waste generated substantial methane (Figs. 2 and 3) compared to
fresh softwood bedding. This was likely from urine and small par-
ticles of manure that were included with this material.
Straw was tested for methane production potential in the 46 d
batch test (Exp. 3). Straw alone produced similar amounts of meth-
ane, 54 ± 12 L/kg VS (Fig. 3), as horse manure alone, 53 ± 15 L/kg VS
(data not shown), but this value was less than half that reported by
Trioloa et al., 2011 for straw alone (289.5 ± 17.5 mL/g VS). Never-
theless, use of straw as bedding would result in higher production
of methane from stall waste than use of softwood-based bedding,
whose presence dilutes the methane potential.
3.2. Determination of maximum methane production potential, Gpot
The Gpot estimated from a fit of Eq. 1 to data from three batch
horse manure experiments are shown in Fig. 4 and Table 3. The Gpot
of 68, 191 and 273 mL methane/g VS indicated heterogeneity be-
tween different batches of manure or perhaps differences in the
effectiveness of inoculation. All other batch test data for the 46,
59 and 79 day runs were fit with Eq. 1 as described in Section
300
Cummulative Methane Production (mL/g VS)
250
200
150
100
50
0
0 50 100
Time (Days)
Fig. 4. Estimates of Gpot for three different methane production potential tests from
horse manure alone. Symbols are averages of replicate bottles and the errors bars
are one standard deviation. The lines indicate the model derived values by a fit to
the average of the replicate data. [s, 46 day batch run, ——————— model fit; h,
59 day batch run, ——— model fit; and }, 79 day batch run, — — — model fit].
Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
5
2.5, to estimate Gpot for different mixtures of waste materials (Ta-
ble 3). As expected, Gpot decreased as an increasing ratio of soft-
wood bedding was added to the mixture. For example, a 1:1 (on
a VS basis) mixture of horse manure and fresh softwood chip bed-
ding yielded a Gpot of 91 ± 17 mL/g VS for the 79 d batch run, or 48%
of the Gpot of 191 mL/g VS estimated for manure alone. In contrast,
Kusch et al. (2008) reported a Gpot of 277 mL/g VS for straw-based
stable waste, with the exact proportion of straw to manure not re-
ported, but reflecting real conditions in a working facility.
3.3. Effect of increasing bedding to manure ratios on methane
production
As seen in Fig. 5a, fresh softwood bedding did not inhibit meth-
ane production relative to controls over a wide range of loadings
from 0.01:1 to 4:1 g softwood bedding VS to g horse manure VS.
The average of the ratios of methane production in treatments
receiving softwood bedding relative to the controls with horse
manure only was 0.84 ± 0.24 (average ± one standard deviation),
i.e. less than the 1:1 ratio expected if no inhibition occurred, since
the bedding itself was scarcely biodegradable (Fig. 3). However,
there was no indication of a dose response wherein higher ratios
of softwood bedding resulted in successively less methane produc-
tion relative to controls. Further, ANOVA of data as a whole from all
treatments indicated that there was no statistically significant dif-
ference between the methane production relative to the control
(p = 0.36), nor was there a statistically significant difference
between groups of treatments receiving softwood bedding
(p = 0.19). Taken together, these results indicated that regardless
of the amount of fresh softwood bedding present in the waste mix-
ture, no inhibition was observed. We earlier observed a decrease in
methane production when complete mix reactors were switched
from horse manure alone to stall waste with softwood bedding
(Wartell et al., 2008). It is possible that that decrease was caused
by sudden exposure of the microbial community to the wood.
Results of similar comparative analysis for softwood pellets and
straw are shown in Fig. 5b and c. No apparent effect on methane
production from horse manure was noted regardless of the relative
amount of softwood pellets added. The average of the ratios of
methane production in treatments receiving softwood pellets rela-
tive to the controls with horse manure only was 1.2 ± 0.2 (aver-
age ± one standard deviation), with no decreasing trend in
methane production at higher ratios. Indeed, some increase in
methane production was seen at the higher ratio of 4:1 softwood
pellet VS: horse manure VS.
Used softwood bedding that was manually separated from stall
waste was also tested for inhibition of methanogenesis at ratios of
0.25:1, 0.5:1, 1:1, 2:1, and 4:1 g bedding VS to g horse manure VS.
Results from this test also indicated no inhibition of methanogen-
esis, regardless of the amount added (data not shown). Moreover,
this material exhibited substantial methane production (Figs. 2
and 3) with an estimated Gpot of 71 ± 22 L methane per kg VS (Ta-
ble 3).
Straw bedding contributed substantially to methane production
(Fig. 5c) as expected based on its high biodegradability, increasing
methane production nearly linearly up to a 4:1 ratio of bedding to
horse manure. It is possible that the highest ratio tested (4:1) did
not produce as much methane as expected because of need for fur-
ther incubation time, or perhaps acidification of the system at this
150 higher loading. Indeed, as mentioned previously, the methane pro-
duction we observed from the 46 day batch study (Exp. 3) for straw
alone (53 ± 11 mL/g VS) or 1:1 straw – horse manure mixtures
(68.5 ± 29 mL/g VS) were less than half that reported by Trioloa
et al., 2011 for straw alone (289.5 ± 17.5 mL/g VS), by Kusch et al.
(2008) for straw – manure stall waste (170 mL/g VS) and by Cui
et al. (2011) for spent straw from horse stalls (150 mL/g VS). This
6 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx

Table 3
Gpot, methane production potential, as t ? 1 for horse manure, stall waste mixtures and bedding types expressed on the basis of the total (horse manure plus bedding) volatile
solids added.

Waste Bedding VS: Manure VS (g:g) Gpot (mL CH4/g VS added) Reference
Horse manure
a
79 d na (bn = 3) 191 This study
59 d na (n = 3) 273
46 d na (n = 2) 68
Horse manure mixed with used softwood chip bedding (79 d) 0.25 (n = 3) 148 ± 28 This study
0.5 (n = 3) 126 ± 22
1 (n = 3) 124 ± 1
2 (n = 3) 91 ± 6
4 (n = 3) 74 ± 2
Horse manure mixed with fresh softwood bedding (59 d) 0.01 (n = 3) 291 ± 108 This study
0.025 (n = 3) 237 ± 20
0.05 (n = 3) 197 ± 32
0.1 (n = 3) 243 ± 16
0.5 (n = 2) 138 ± 10
Horse Manure Mixed with Fresh Softwood Bedding (79 d) 1 (n = 3) 91 ± 17
c
Softwood Fresh Chips nd This study
Bedding only Fresh Pelleted nd
Manually Separated (Used) Chips (n = 9) 71 ± 22
Horse Manure – Straw Bedding Not reported 277 ± 5 Kusch et al. (2008)
a
na, not applicable
b
n is the number of replicates analyzed
c
nd – not determined.

(a) (b)
1.5 Softwood chips 59 d 1.5
Methane Production Relative to Control

Methane Production Relative to Control

Softwood chips 40 d
Softwood chips 46 d

1 1
(mL/mL)

(mL/mL)

0.5 0.5

0
0
0 0.5 1 2 4
0 0.01 0.05 0.1 0.25 0.5 1 2 4
Ratio of Bedding to Horse Manure
Ratio of Bedding to Horse Manure
(g bedding VS / g horse manure VS) (g bedding VS / g horse manure VS)

(c) 5
Methane Production Relative to Control

3
(mL/mL)

0
0 0.5 1 2 4
Ratio of Bedding to Horse Manure
(g bedding VS / g horse manure VS)

Fig. 5. Methane production relative to manure only controls (mL/mL) from batch anaerobic reactors amended with (a) horse manure and different ratios of fresh softwood
chip bedding during three separate experiments; (b) horse manure and different ratios of fresh pelleted softwood bedding; and (c) horse manure and different ratios of fresh
straw bedding. Ratios are expressed on bedding volatile solids (VS) to horse manure VS basis. Values are averages of triplicate bottles and error bars are one standard
deviation.

may have been caused by inadequate inoculation, acidification or indicating that use of straw as bedding would add to energy pro-
inadequate incubation time. However the methane production duction from stall waste as suggested by Møller et al. (2003), rather
from horse manure alone (53 ± 15 mL/g VS) during Exp. 3 was than dilute it, as softwood bedding would. Thus, in considering
similar to that of straw alone and manure-straw mixtures, anaerobic digestion of equine waste, choice of stall bedding is a

Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx 7

60 57 days and Digester #2 56 days). No mass balance on VS loss

Digester #1 Re-inoculated was performed. Operation of Digester #2 was initially less stable
Digester #2 than Digester #1 because of a problem with the temperature sen-
Cumulative Methane (L/kg VS)

50
sor. The reactor temperature reached approximately 46 °C several
times prior to switching to a new sensor on Day 7. On Days 50 (Di-
40
gester #1) and 43 (Digester #2), each digester was reinoculated
and methane production consequently increased in both reactors.
30 The maximum methane content of the biogas approached 50%
for Digester #1 between Days 21 and 37, but then a slow but grad-
20 ual decline in methane content occurred between Days 37 and 47
Re-inoculated
(data not shown). For Digester #2, methane concentrations ap-
10 proached only 45% between Days 12–21, but then a slow but grad-
ual decline in methane content occurred between Days 21 and 37
(data not shown). Re-inoculation increased the methane content of
0
0 5 10 15 20 25 30 35 40 45 50 55 60 65 the biogas in both reactors, but these did not exceed 50%. The rel-
Time (days) atively low methane content could be a result of the low moisture
content and/or inadequate mixing of the inoculum and leachate in
Fig. 6. Cumulative methane production (L) from replicate 125 L high solids batch the pile (Martin, 1999). Further, compared to Kusch et al. (2008)
digesters loaded with stall waste from horses bedded on softwood pellets. [j,
Digester #1; h, Digester #2].
and Cui et al. (2011) proportionally less inoculum was used in
our study and may not have been sufficient for effective digestion.

critical factor in the overall energy potential and thus economics of 3.5. Implications
the digestion process, in addition to existing considerations for
horse health, safety and comfort. Kusch et al. (2008) examined pretreatment and inoculation
methods using a variety of solid state reactor configurations for
3.4. Solid state digestion of stall waste in 125 L reactors digestion of stall waste consisting of manure and straw bedding,
which has an equal or greater methane yield per unit weight than
The cumulative biogas produced from actual softwood pellet- the manure itself (Møller et al. (2003); Trioloa et al., 2011). The Gpot
based stall waste was 1320 L for Digester #1 and 1097 L for Diges- of the manure-straw waste was reported to be 277 mL methane/g
ter #2 after 63 and 56 days, respectively (data not shown). The cor- VS (Kusch et al., 2008) with a methane yield of 170 mL/g VS after
responding cumulative methane production from these static piles 40 days of digestion. Cui et al. (2011) examined methane produc-
of waste that were originally loaded with 9.8 kg VS is shown in tion from spent straw removed from horse stalls and reported
Fig. 6. The average total methane produced was 29 ± 5.9 L/kg VS methane production of 150 mL methane/g VS after approximately
after 1 month and 44.8 ± 3 L/kg VS after 2 months (Digester #1 30 days. The methane production values reported by Kusch et al.

Table 4
Energy (1  105 kJ/ metric ton wet wt) generated during 20 to 40 d incubation for selected wastes.

Material/experiment/incubation time used for analysis Bedding g VS: horse manure g VS

a
na 0.25 0.5 1 2 4
Horse manure
Exp 1, 21 d 8.09 ± 2.1 na na na na na
Exp 2, 40 d 8.45 ± 5.42 na na na na na
Exp 3, 34 d 3.11 ± 0.92 na na na na na
Exp 4, 32 d 5.54 ± 0.89 na na na na na
Exp 5, 33 d 3.44 ± 0.89 na na na na na
Horse Manure + Fresh Softwood Chips
Exp 1, 21 d na 9.97 ± 0.68 5.63 ± 2.2 na na na
Exp 2, 40 d na 6.25 ± 1.8 6.57 ± 1.14 na na na
Exp 3, 34 d na na 1.87 ± 0.69 1.69 ± 0.39 1.43 ± 0.96 1.34 ± 0.05
Exp 4, 32 d na na na 3.91 ± 0.47 na na
Exp 5, 33 d na 2.08 ± 0.45 1.67 ± 0.76 1.71 ± 0.4 1.79 ± 0.17 na
Fresh Softwood Chip Bedding
Exp 1, 21 d 0.97 ± 0.28 na na na na na
Exp 2, 40 d 0.86 ± 0.28 na na na na na
Exp 3, 34 d 0.32 ± 0.16 na na na na
Exp 4, 32 d 0.11 ± 0.03 na na na na
Exp 5, 33 d 0.73 ± 0.11 na na na na na
Horse Manure + Used Softwood Bedding
Exp 4, 32 d na 3.73 ± 0.63 3.36 ± 0.18 3.76 ± 0.41 3.11 ± 0.72 3.03 ± 0.45
Used Softwood Bedding
Exp 4, 32 d 2.34 ± 0.46 na na na na na
Softwood Pellets + Manure
Exp 3, 34 d na na 2.54 ± 0.49 2.22 ± 0.02 2.10 ± 0.51 2.2 ± 0.15
Straw + Manure
Exp 3, 34 d na na 3.87 ± 1.25 5.83 ± 2.45 8.44 ± 0.13 5.22 ± 0.37
Farm stall waste with softwood pellet bedding
b
125 L digesters, 33 d na na na 3.21 ± 0.66 na na
a
na, not applicable.
b
The estimated bedding VS: horse manure VS was 1:1–2:1 for this material.

Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
8 B.A. Wartell et al. / Bioresource Technology xxx (2012) xxx–xxx
(2008) and Cui et al. (2011) are similar to the upper values deter-
mined in this study for horse manure alone and for horse manure
with low proportions of softwood bedding. The highest methane
potentials we estimated for horse manure, 191 and 273 mL/g VS
(Table 3), were lower than those experimental or modeled meth-
ane production potentials reported for swine manure:
356 ± 28 mL/g VS (Møller et al., 2003), 280 to 360 mL/g VS (Liu
et al., 2009) and 213.8 ± 9.2–417.2 ± 11.8 mL/g VS (Trioloa et al.,
2011). The horse manure values were similar to those reported
for cattle manure: 148 ± 41 mL/g VS (Møller et al., 2003), 150–
240 mL/g VS (Liu et al., 2009) and 197.0 ± 3.3–223.6 ± 3.7 mL/g
VS (Trioloa et al., 2011). Further, these values compare favorably
to methane potentials reported for many types of crop wastes
(for an extensive review see Nallathambi Gunaseelan, 1997) indi-
cating that horse waste is a favorable feedstock for anaerobic
digestion.
The softwood based bedding was barely degradable under
anaerobic conditions (Fig. 3). Wood particles could potentially
cause mechanical clogging of digesters where piping or pumping
is utilized, and require larger reactor volumes to accommodate this
relatively non-digestable material. Thus, one consideration for
anaerobically digesting equine stall waste that contains softwood
chip bedding would be to mechanically separate the bedding from
the waste and digest only the manure. The separated bedding
could be used in composting or other applications. Our data sug-
gest that separation of the bedding from the manure prior to recov-
ery of bioenergy would result in a substantial loss of recoverable
energy since the Gpot for manually separated bedding was
71 ± 22 L methane per kg VS and that for horse manure alone in
that specific batch was 191 L methane per kg VS (Table 3). Notably,
Cui et al. (2011) reported that used straw bedding produced up to
150 mL methane/g VS over 30 days, more than twice what we esti-
mated as the potential for the used wood chips. The methane pro-
duced from the manually separated, used softwood bedding was
likely contributed by digestion of small manure remnants that ad-
hered to the wood particles or urine that had soaked into the wood.
It is also possible that some breakdown of the bedding itself oc-
curred, after weathering in stalls.
The energy potential expressed as kJ/metric ton wet weight of
horse manure or various horse manure – bedding mixtures were
estimated (Table 4) using cumulative methane production over
20–40 days of batch digestion to reflect a time-scale that might
be used in full scale operations. There was substantial variation
in the methane produced. However, if one considers the upper
and lower energy production values for a 1:1 (VS basis) mixture
of softwood chip bedding to manure (i.e., 1.7–3.9 kJ/metric ton)
and that one horse produces approximately 12 metric tons wet
weight of stall waste per year (Krogmann et al., 2006; Wheeler
and Zajaczkowski, 2002), this amounts to an energy production
of 2–4.7 GJ (106 kJ) generated per horse per year. Certainly, some
of this energy would be lost, depending upon the conversion sys-
tem used to recover energy from the methane. In regions with high
horse densities, stall waste digestion could supply a portion of
renewable energy needs. Further investigation of the economics
of this process and the need for transport and collective digestion
to include small equine operators should be further investigated.
Based on the results from the 125 L static pile reactors, more work
on optimization of reactor operation is needed to increase the effi-
ciency of bioenergy recovery from this material.
4. Conclusions
The maximum methane production potential, Gpot, estimated
for horse manure alone was 273, 191 and 68 L methane per kg
VS in separate batch tests. While straw bedding contributed as
Please cite this article in press as: Wartell, B.A., et al. Methane production from horse manure and stall waste with softwood bedding. Bioresour. Technol.
(2012), doi:10.1016/j.biortech.2012.02.012
much or more methane potential as the manure, softwood-based
bedding had a substantial dilution effect on potential energy
production from equine stall waste. There was no inhibition of
methanogenesis by softwood bedding. The methane production
potential, Gpot, for manually separated bedding was 71 ± 22 L
methane per kg VS, indicating that substantial loss of energy could
occur as a result of removing the softwood bedding prior to
digestion.
Acknowledgements
Funding was provided by the Rutgers Equine Science Center’s
State Equine Initiative, the New Jersey Agricultural Experiment
Station, and by a USDA CSREES Region 2 - NJ Water Resources
Research Institute Undergraduate Student Stipend Program grant.
Stall waste, softwood bedding and horse manure was obtained
from Michael Fennell and Joanna Powell of the New Jersey Agricul-
tural Experiment Station (NJAES) Animal Care Program on the Cook
Campus of Rutgers University, New Brunswick, NJ. We thank
Professor Uta Krogmann for providing the 125 L reactors and Ross
Hull for technical assistance. We thank Tara Bowles and Oxbow
Stables, Inc. for providing real stall waste and information about
stall bedding and cleaning. Finally, we gratefully acknowledge
the assistance and support of Professor Karyn Malinowski,
Director, and the staff of the Rutgers Equine Science Center.
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