Agricultural Wastes 6 (1983) 175-191

Thermophilic and Mesophilic Anaerobic Fermentation of

Swine Manure*

Andrew G. Hashimoto

Roman L. Hruska US Meat Animal Research Center,

Agricultural Research Service, US Department of Agriculture,
Clay Center, Nebraska 68933, USA

A BSTRA CT

The effects of temperature (35 ° and 55°C), influent Volatile Solids (VS)
concentration (S O = 50.4 and 62.5 kg VS m- 3) and hydraulic retention
time (HRT = 5, 10, 15 and 25 days) on methane (CH4) production from
swine manure were evaluated using 3-dm 3 laboratory scale fermentors.
The highest CH 4 production rate achieved was 3.12m 3 CH 4 per m 3
fermentor per day at 55°C, 5-day H R T and S O = 50.4 kg VS m- 3. Batch
fermentations showed an ultimate CH 4 yieM (Bo) of 0.49 m 3 C H 4 kg VS
fed. Extreme fermentation instability was experienced by the thermo-
philic ( 55°C) fermentors at 25-day H R T and S O = 62.5 kg VS m - 3. This
instability was attributed to free-ammonia and influent-substrate
inhibitions. After a significant period for adaptation (over 170 days), the
thermophilic fermentors adapted to the high free-ammonia concentra-
tions and operated satisfactorily.

INTRODUCTION

T h e digestibility o f s w i n e m a n u r e , as m e a s u r e d b y the u l t i m a t e m e t h a n e
( C H 4 ) yield (B 0 e x p r e s s e d as m 3 C H 4 p e r k g VS fed at infinite r e t e n t i o n

* Mention of commercial or proprietary products in this paper does not constitute

recommendation or endorsement of these products by the US Department of Agriculture.
175
176 Andrew G. Hashimoto

time), is significantly higher (0.50 + 0.05 m a CH 4 per kg VS fed) than the

B o for beef cattle manure (0.35 + 0.05m a CH 4 per kg VS fed) or dairy
cattle manure (0-20 + 0.05 m 3 CH 4 per kg VS fed) (Hashimoto et al.,
1981a). However, about four times higher CH 4 production rates have
been achieved by fermenting cattle manure compared with swine manure
(Hashimoto et al., 1981a). The primary reason for the higher CH 4
production rates from cattle manure was the higher loading rates used for
the cattle manure fermentations (16 to 20kg VS m - 3 at thermophilic
conditions). Design, or optimum, swine fermentor loading rates were
reported to be: 4 k g Total Solids (TS) m - 3 day -1 at 30 to 35°C (van
Velsen et al., 1979); 4 k g VS m -3 day -1 at 35°C (Fischer et al., 1979);
3.84kg VS m - 3 day -1 at 35°C (Gramms et al., 1971); 2.24kg VS m - 3
d a y - 1 at 35 °C (Sievers & Brune, 1978) and 2.5 kg VS m - 3 d a y - 1 at 35 °C
(Huang et al., 1978). These loading rates were for mesophilic
fermentations and were a quarter to a fifth the loading rate achieved for
cattle manure fermented at thermophilic temperatures. Hashimoto
(1981) showed that, for beef-cattle manure, much higher loading rates
could be used at thermophilic temperatures (about 20 kg VS m - 3 day - 1)
compared with mesophilic temperatures (about 7kg VS m - 3 d a y - l ) .
However, van Velsen et al. (1979) showed that, for swine manure at the
same loading rate and hydraulic retention time (HRT), mesophilic
fermentations produced about 259/0 more CH 4 than thermophilic
fermentations.
A m m o n i a inhibition is considered to be the primary factor limiting the
loading rate to swine manure fermentors. McCarty & McKinney (1961)
showed that ammonia toxicity occurs at a free-ammonia concentration of
0"15 kg m-3. McCarty (1964) later reported general guidelines stating
that ammonia inhibition occurred at ammonia (free ammonia +
ammonium) concentrations between 1.5 and 3.0kg N m -a at pH above
7.4 to 7.6, and ammonia toxicity occurred at concentrations above 3.0 kg
N m - 3 at all pH levels. Hobson & Shaw (1976) confirmed these
guidelines, showing that Methanobacterium f o r m i c i c u m was partially
inhibited in producing CH 4 at an ammonia concentration of 2.5 kg N
m - a and pH of 7.1, and was completely inhibited at an ammonia
concentration of 3.3 kg N m - a. Similarly, Kroeker et al. (1979) using
urea and acetic acid, concluded that CH 4 production was progressively
inhibited as the ammonia concentration increased above 2 kg N m - 3 ,
although complete cessation of CH 4 production did not occur even at an
ammonia concentration of 7 kg N m - a . However, several researchers
 Anaerobic fermentation of swine manure 177

have recently reported stable fermentor operation at ammonia concentra-

tions in excess of 2 k g N m -3 (Gramms et al., 1971; Lapp et al., 1975;
Fischer et al., 1979; Stevens & Schulte, 1979; Converse et al., 1981).
The stable operation of mesophilic, anaerobic fermentors at ammonia
concentrations greater than 2 kg N m - 3 was attributed to the acclimation
of methanogens to high a m m o n i a concentrations. Inhibition of CH4
production occurred at about 1.7 kg N m - 3, but this inhibition was only
temporary when methanogenic organisms were allowed to acclimate, and
organisms acclimated to 1.7 kg N m - a were also acclimated to ammonia
concentrations up to 2-7kg N m -3 (van Velsen, 1979a). Further,
organisms acclimated to 2.4 kg N m - 3 were also acclimated to ammonia
concentrations up to 3.1 kg N m - 3, and methanogens present in sewage
sludge acclimated to ammonia concentrations as high as 5 kg N m - 3 (van
Velsen, 1979b). However, van Velsen et al. (1979) reported that ammonia
acclimation was not apparent for thermophilic (55°C) fermentation of
swine manure.
Swine manure fermentations at 55 °C (van Velsen et al., 1979) and 40 °C
(Stevens & Schulte, 1979) resulted in lower gas production than
fermentations at 35°C. Both references attributed the lower gas
production to free-ammonia inhibition. Since the free-ammonia con-
centration increases as temperature and pH increase, it is expected that
more free-ammonia would be present at higher temperatures. For
example, at a total ammonia concentration of 3.5 kg N m - 3 and a pH of
8, the free-ammonia concentration would be 0.97kg N m -3 at 55°C
compared with 0.26kg N m -3 at 35°C (van Velsen et al., 1979).
This study was initiated to study the effects of temperature (35 ° and
55 °C), influent VS concentration and hydraulic retention time on CH4
production from swine manure.

METHODS

Batch and daily fed (once a day) fermentors were used in this study. The
batch fermentors were used to determine B 0 of the swine manure and the
daily fed fermentors were used to evaluate the effects of temperature,
influent concentration and H R T on CH 4 production. The batch and daily
fed fermentors were 4-dm 3 aspirator bottles with working volumes of
3 dm 3, as described previously (Hashimoto et al., 1981 b). The fermentors
were mixed using a platform shaker rotating at 140 revolutions per
178 Andrew G. Hashimoto

minute. The fermentors were housed in a constant temperature chamber

and temperatures were controlled within 1 °C using heating tapes
wrapped around the fermentors.
Swine manure (faeces and urine) was collected from pigs (weighing 80
to 90 kg) confined in metabolism stalls and fed a ration consisting of
82" 1 ~o corn (No. 2 yellow dent), 14.0 ~o soybean meal, 2.4 ~o dicalcium
phosphate, 0"5~o ground limestone, 0 . 4 ~ iodised salt, 0"4~o vitamin
premix and 0.4 ~o trace minerals. This ration contained approximately
14 ~o protein, 0.8 ~o calcium, 0.8 ~o phosphorus and metabolisable energy
of 13 MJ k g - 1. Antibiotics were not used in the ration.
One-day old manure was diluted to about 14 ~o dry matter, placed in
plastic bags (about 5 kg per bag) and frozen at - 20 °C. Periodically, a bag
of manure was removed from the freezer, allowed to thaw overnight in a
regrigerator, and appropriate amounts of slurry were weighed into
polyethylene bottles. The bottles were then refrozen until used. Before
use, the bottles were placed in a refrigerator to thaw overnight, and the
slurry was diluted with hot tap water to the desired VS concentration and
volume.
Eight batch fermentors were started by placing 1 dm 3 ofinoculum from
stable fermentors operating at 35 ° and 55°C, and l dm 3 of mineral
solution (Hashimoto et al., 1981 b) in each fermentor. Four fermentors
were maintained at 55°C and four at 35°C for 21 days to allow
temperature equilibration and utilisation of substrate contained in the
inoculum. Two of the fermentors at each temperature were each fed
manure containing 25 g VS in dm 3 over a period of 7 days. The other two
fermentors at each temperature were each fed I dm 3 of the mineral
solution and served as controls. These batch fermentors were operated for
146 days, during which gas volume and composition (CH 4 and CO2) were
measured periodically. At the end of the study, the total volume of CH 4
produced, including the CH 4 present in the fermentor headspace, was
calculated. B 0 was calculated by subtracting the total CH 4 produced in
the control from that produced in the manure fermentors and dividing by
25 g VS.
The daily fed fermentors (two fermentors at 35 °C and 2 at 55 °C) were
started by placing 3 dm 3 of acclimated inoculum in each fermentor. The
fermentors were fed manure slurries containing 62.5 kg VS m - 3 at 25-day
HRT. Steady state was assumed after four volume turnovers, and C H 4
production and effluent characteristics were analysed for five consecutive
days. The influent concentration was then reduced to 50.4 kg VS m - 3 and
 Anaerobic fermentation of swine manure 179

the HRT to 15 days. Table 1 shows the composition of the manure fed to
the fermentors. The same procedure as outlined above was continued
until steady-state data were obtained at 10 and 5 days HRT. However,
only one fermentor was operated at 35°C and 5-day HRT because
insufficient manure was available near the end of the study.
Slurries fed and withdrawn from the fermentors were analysed for
Total Solids (TS), Volatile Solids (VS), Fixed Solids (FS), ammonia

TABLE 1
C o m p o s i t i o n of Swine M a n u r e Fed to F e r m e n t o r s

Constituent" Approximate VS concentration ( k g m - 3)

60 50

T o t a l Solids 79.6 + 5.8 63'6 + 5.5

Volatile Solids 62-5 ___5.2 50-4 ___4.6
Chemical Oxygen D e m a n d 77.4 ___7.1 52.1 + 9.2
Volatile Acids 7.25 _ 0.50 2.95 _ 0.69
pH 6.48 + 0.17 6.57 + 0.26
Alkalinity 11.2 -I- 0.8 7.92 _ 0.56
Ammonia-N 1-75 + 0.08 1-15 + 0.11
Kjeldahl-N 4.58 + 0.30 3-37 + 0.37
Cellulose 6.4 5.2
Hemicellulose 9.8 7.9
Lignin 1.4 1.1
Calcium 2.4 1-9
Phosphorus 2.2 1.8
Potassium 1.9 1.5
G r o s s Energy b 1.6 1.2

" Expressed as kg m - 3 except for p H a n d gross energy.

b Expressed as G J m - 3

(distillation method), chemical oxygen demand (COD), alkalinity (to

pH 3.7), pH and Total Volatile Acids (TVA as acetic acid, silicic acid
method) using standard methods for wastewater analyses (APHA, 1975).
Kjeldahl nitrogen was determined as described by Wael & Gehrke (1975).
Cellulose, hemicellulose, lignin, calcium, phosphorus, potassium and
gross energy were determined by published procedures (AOAC, 1975).
Individual volatile fatty acids (VFA, including acetic, propionic, butyric,
iso-butyric, valeric, iso-valeric, caproic and iso-caproic acids) were
180 Andrew G. Hashimoto

measured by gas chromatography as described previously (Hashimoto et

al., 1981b).
The free-ammonia concentration was calculated using the following
equation:
[NH3] = [T-NH3]/(1 + [H +]/Ka) (1)
Where [NH3] = free-ammonia concentration (kg m - 3),
[T-NH3] = total-ammonia concentration (kg m - 3),
[H ÷ ]~= hydrogen ion concentration (moles litre-1),
KR = acid ionisation constant of ammonia (moles litre- 1).
Values for K a at 35 ° and 55 °C were calculated using the values for the
dissociation constants (Kb) of aqueous ammonia and the ionisation
constants (Kw) for water given in the Handbook o f Chemistry and Phys&s
(CRC, 1968). K a was calculated to be 11.295 moles litre-1 at 35 °C and
approximately 38.29 moles litre-1 at 55 °C by extrapolation.
Biogas produced in the fermentors was collected in gas-impermeable
bags and analysed for gas volume and CH 4 concentrations. The gas
volumes were meausred by a solution-displacement method, and the CH 4
content was measured by gas chromatography as described previously
(Hashimoto et al., 1981b). Data were analysed by least-squares
procedures outlined by Harvey (1975). The main effects were temperature
and HRT.

RESULTS

B o for the 35 °C batch fermentors were 0.490 and 0-503 m 3 CH 4 per kg VS

fed and 0.464m 3 per kg VS fed for the 55 °C fermentor. The other batch
fermentor at 55 °C developed a leak in the gas collection bag; therefore, a
replicate at 55 °C was not obtained. Since fermentation temperature does
not affect the B o of a given substrate (Hashimoto et al., 1981b), the mean
B o was calculated to be 0.49m 3 CH 4 per kg VS fed, with a standard
deviation of 0.02 m 3 CH4 per kg VS fed. This B o value for swine manure is
within the range of 0.50 _ 0.05 m 3 CH 4 per kg VS fed reported previously
(Hashimoto et al., 1981a).
The original experimental design called for the fermentors to receive ca.
60kg VS m -3 at 25-, 15-, 10- and 5-day HRT. However, stable
fermentation could not be achieved by the 55 °C fermentors at 25-day
HRT. Therefore, the influent concentration was reduced to ca. 50 kg VS
m - 3 for the trials at 15-, 10- and 5-day HRT.
 Anaerobic fermentation o f swine manure 181

Figures 1 and 2 show the changes in TVA and methane production rate
(? v, m 3 CH 4 per m 3 fermentor per day), respectively, during the operation
of the fermentors at an influent concentration of 62-5 kg VS m - 3 and 25-
day HRT. The TVA for the 35 °C fermentors remained fairly constant at
about 1 kg m-3 throughout the trial. In contrast, the TVA in the 55 °C
fermentors continued to increase throughout the trial. The decrease in
TVA after day 38 resulted from the addition of 1 dm 3 of active,

tD

I I
g, s, ,o6
TIME (DAYS)
Fig. 1. Change in Total Volatile Acids (TVA) during fermentation of swine manure
at an influent Volatile Solids content of 62.5 k g m - 3 and at 35 ° ( I ) and 55°C ( 0 ) .

thermophilic inoculum. This was effective in temporarily reducing the

TVA from c a . 4 k g m -3 to ca. 2 k g m -3. However, within 25 days
(1 HRT), the TVA was above 4 kg m - 3 and continued to increase to
ca. 9 kg m -3 between days 104 and 108.
Individual VFA profiles are shown for the 55 o (Fig. 2) and 35 °C (Fig. 3)
fermentors. Figure 2 shows that on day 28, acetic and propionic acids
were above 1 kg m - 3 and significant levels of butyric (ca. 0.3 kg m-3) and
valeric (ca. 0 . 4 k g m -3) acids were present. After the 55°C fermentors
were reinoculated on day 28, acetic, butyric and valeric acids decreased
while propionic acid continued to gradually increase. Acetic, butyric and
valeric acids remained relatively constant until day 55, after which all
three acids began to increase. After day 69, caproic acid began to
gradually increase and, after day 85, acetic, propionic, butyric and valeric
182 Andrew G. Hashimoto

t3
Z
0
2
<~
IJ.
>

Cl j
w
20 40 60 80 I00
TIME (DAYS]

Fig. 2. Change in individual Volatile Fatty Acids ~(VFA) during fermentation of swine
manure at an influent Volatile Solids concentration of 62.5 kg m - ~ and at 55 °C
(C), acetic; 0 , propionic; I , butyric; A , valeric and O, caproic).

>

(.9

d
Z
O
t.3

20 4O 60 80 I00
TIME (DAYS)

Fig. 3. Change in individual Volatile Fatty Acids (VFA) during fermentation of

swine manure at an influent Volatile Solids concentration of 62.5 kg m - 3 and at 35 °C
( O , acetic; 0 , propionic).
 1.2

.8,
%
v

I1.

ii
IOe
TIME (DAYS)
Fig. 4. Change in m e t h a n e p r o d u c t i o n rate during fermentation o f swine m a n u r e at
an influent Volatile Solids c o n c e n t r a t i o n o f 62.5kg VS per m 3 and at 35 ° ( I ) and
55 °C (O).

TABLE 2
Effluent Characteristics and M e t h a n e P r o d u c t i o n o f Swine M a n u r e F e r m e n t o r s Operated
at 25-Day Hydraulic Retention Time and Influent C o n c e n t r a t i o n o f 62.5 kgVS per m 3

Parameter Temperature

55°C 35°C

Total Solids (kg m - 3) 42'9 _ 4"5 a 29"2 + 0' 1"

Volatile Solids (kg m - 3) 30"4 + 4"M 18'9 + 0'3 a
Chemical Oxygen D e m a n d (kg m - 3) 55"1 + 1"7b 30-3 + 1'2 b
Kjeldahl-N (kg m - 3) 4"26 _ 0"21 b 4'20 ___0"06 b
A m m o n i a - N (kg m - 3) 2"94 ___0"05 b 2"56 + 0'01 b
Total Volatile Acids (kg m - 3) 8"74 + 0"92 b 1"15 + 0"01 b
Alkalinity (kg m - 3) 16'2 + 1"4b 13"9 + 0'1 b
pH 7"63 +0"01 b 7'79 ___0'03 b
Methane-(%) 41 "7 + 10"7 b 60"9 ___0'8 b
Methane production
m 3 per m 3 per day 0'26 ___0"12 b 1'04 ___0'02 b
m 3 per kg VS fed 0"10 + 0"05 b 0'42 + 0'01 b
m 3 per kg VS used 0"20 + 0"10 b 0"59 + 0"01 b

a D a t a presented as m e a n _+ one s t a n d a r d deviation o f two fermentors per treatment (one

observation per fermentor).
b D a t a presented as mean _+ one s t a n d a r d deviation o f two fermentors per treatment (five
observations per fermentor).
 184 Andrew G. Hashimoto

acids increased sharply. In contrast, acetic acid was the only predominant
fatty acid in the 35 °C fermentors (Fig. 3), indicating stable anaerobic
fermentation.
The CH4 production rate (Fig. 4) from the 35 °C fermentors increased
until day 85, then remained fairly constant at ca. 1 m 3 c n 4 per m 3
fermentor per day. In contrast, the CH 4 production rate from the 55 °C
fermentors remained fairly constant (ca. 0-9 m 3 per m 3 fermentor per day)
between days 28 and 76, then decreased sharply to ca. 0.3 m 3 CH 4 per m 3
fermentor per day between 94 and 108. The sharp drop in CH 4
production rate began when the TVA exceeded 4 kg m - 3. The increase in
TVA with time was indicative of fermentation instability. The free-
ammonia concentration was 0.41 and 0.17 kg N m - 3 for the 55 ° and 35 °C
fermentors, respectively. Table 2 summarises the effluent characteristics
TABLE 3
Effluent Characteristics a n d M e t h a n e P r o d u c t i o n o f Swine M a n u r e F e r m e n t o r s Operated
at 55°C a n d Influent C o n c e n t r a t i o n of 50.4kg VS per m 3

Parameter Hydraulic retention time a

15 days 10 days 5 days

T o t a l Solids ( k g m -3) 33.2+2.1 29.3+0.1 36.1 +2.3

Volatile Solids ( k g m -3) 21.7 ___0.8 19.2 + 0.7 25.1 + 1.7
Chemical Oxygen Demandl ( k g m -3) 31.5 _ 0.4 28.4 _ 2'6 33.8 _ 0'5
Kjeldahl-N (kg m - 3) 3.78 +__0'05 3.29 + 0.16 3-34 + 0.05
A m m o n i a - N (kg m - 3) 2.06 _ 0'08 1-86 + 0.25 1-77 + 0' 10
Total Volatile Acids (kg m - 3) 1.90 + 0.11 1.22 + 0.14 1.88 ___0.52
Volatile F a t t y Acids (g m - 3)
Acetic 271 +__17 534 -I- 95 439 _ 148
Propionic 1 240 + 47 502 + 56 1 198 ___257
Butyric 33 + 1 6 _ 8 42 + 1
Valeric 18 + 0 42 _ 1 178 _ 88
Caproic 0 0 6+ 8
Alkalinity ( k g m -3) 12.1 ___0.4 11.1 + 0 . 2 10.5 -I-0-7
pH 8-01 _ 0 . 0 0 7.80+0.10 7.72+0.03
M e t h a n e (~o) 61.0 _ 0 . 1 61.3 _+0.4 61.1 + 0 . 1
Methane production
m3 per m3 per day 1.45+0.09 1.80+__0.12 3.12+0.15
m 3 per kg VS fed 0.43 + 0.03 0.36 + 0.02 0.31 + 0.10
m 3 per kg VS used 0.74 _ 0.05 0-57 _ 0.03 0.62 _ 0.03

a Datal presented as m e a n + one s t a n d a r d deviation o f two fermentors per t r e a t m e n t (five

observations per fermentor).
 Anaerobic fermentation o f swine manure 185

a n d C H 4 p r o d u c t i o n f r o m the 35 o a n d 55 °C f e r m e n t o r s . T h e s e results
s h o w the instability o f t h e r m o p h i l i c , a n a e r o b i c f e r m e n t a t i o n o f swine
m a n u r e at a n influent c o n c e n t r a t i o n o f 62.5 k g VS m - a a n d 2 5 - d a y H R T ,
w h e r e a s m e s o p h i l i c f e r m e n t a t i o n at the s a m e influent c o n c e n t r a t i o n a n d
H R T was stable.
Because the t h e r m o p h i l i c f e r m e n t o r s were u n s t a b l e , the influent
c o n c e n t r a t i o n was r e d u c e d f r o m 62.5 to 5 0 . 4 k g VS m -3. T h e 5 5 ° C
f e r m e n t o r s were restarted b y r e p l a c i n g 2 d m 3 o f slurry with I d m a o f
i n o c u l u m f r o m the 3 5 ° C f e r m e n t o r s a n d 1 d m a o f i n o c u l u m f r o m a
thermophilic, beef-manure fermentor.
T a b l e s 3 a n d 4 s u m m a r i s e the effluent characteristics a n d C H ,
p r o d u c t i o n f r o m the 55 ° a n d 35 °C f e r m e n t o r s , respectively. T h e results

TABLE 4
Effluent Characteristics and Methane Production from Swine Manure Fermentors
Operated at 35 °C and Influent Concentration of 50.4kg VS per m 3

Parameter Hydraulic retention time

1.5 days a 10 days ~ 5 days b

Total Solids (kg m- 3) 37-0 _ 1.3 36-4 _ 1.3 39-4

Volatile Solids (kgm-3) 25.4 +__1-6 25.1 + 1-3 28.4
Chemical Oxygen Demand (kg m- 3) 31.9 _ 0-7 30.9 _ 0.4 32.1
Kjeldahl-N (kg m- 3) 3.52 + 0.07 3.96 + 0.55 3.29
Ammonia-N (kgm-3) 1.64 _ 0.07 1.70 + 0.02 1.66
Total Volatile Acids (kg m- 3) 0.22 _ 0.01 0.28 + 0.04 0.41
Volatile Fatty Acids (g m- a)
Acetic 52 + 5 113 + 31 186
Propionic 0 0 41
Butyric 0 0 0
Valeric 0 5+ 7 47
Caproic 0 0 0
Alkalinity (kgm -3) 10.9 -t- 0-2 10.3 +0.1 10.2
pH 7-49 _ 0.01 7.43 + 0.02 7.54
Methane (~o) 58.9 _ 0.7 61.8 + 0.4 64.2
Methane production
m s per m a per day 1.22 _ 0.04 1'91 + 0-06 2"69
m 3 per kg VS fed 0"36 + 0-01 0.38 + 0-01 0.26
m s per kg VS used 0.72 + 0.03 0.76 + 0-03 0-60

° Data presented as mean _+one standard deviation of two fermentors per treatment (five
observations per fermentor).
b Data presented as mean of one fermentor (five observations).
186 Andrew G. Hashimoto

shown in Tables 3 and 4 indicate stable fermentation at 35 ° and 55 °C.

Least-squares analysis of the data showed that the 55°C fermentors
produced significantly (P _< 0.05) higher CH 4 production rates, had
higher concentration of ammonia, TVA and pH and lower effluent VS
concentrations than the 35 °C fermentors. The individual VFA from the
55°C fermentors were also significantly higher than from the 35°C
fermentors. The predominant fatty acid from the 35 °C fermentors was
acetic acid, while the predominant acids from the 55 °C fermentors were
propionic and acetic acids.
The free-ammonia concentrations were calculated using eqn. (1) and
data in Tables 3 and 4. At 35 °C, free ammonia concentrations were 0.06,
0-05, and 0-06 kg N m - 3 at 15-, 10-, and 5-day HRT, respectively. At
55 °C, free-ammonia concentrations were 0.58, 0.36 and 0-30 kg N m - 3 at
15-, 10-, and 5-day HRT, respectively.

DISCUSSION

The results of this study have shown fermentation instability at 55 °C, 25-
day HRT and influent concentration of 62.5 kg VS m - 3. No instability
was noted at the same influent concentration and HRT but a fermentation
temperature of 35 °C. However, at an influent concentration of 50.4 kg VS
m -3 and an HRT of 15, 10 and 5 days, instability was not observed at
fermentation temperatures of 55 ° and 35 °C. It was unlikely that high
loading rate caused the unstable fermentation because the loading rates at
50.4 kg VS per m 3 per day were higher than at 62.5 kg per m 3 (3.4, 5.0 and
10.1 kg VS m 3 per day at 50.4 kg VS per m 3 as against 2.5 kg VS per m 3 per
day at 62-5 kg VS per m3).
One probable cause for the instability at 62.5 kg VS per m 3 was the
higher free-ammonia concentration at 55 °C compared with 35 °C (0-41
compared with 0-17 kg N m-3). However, it is not likely that the flee-
ammonia concentration was the only cause of instability since
comparable levels of free-ammonia were experienced at 55 °C and 50.4 kg
VS m -3 (0.58 to 0.30 kg N m -3) with no apparent instability. It is probable
that the high free-ammonia concentration at 55 °C contributed to the
instability at 62.5 kg VS m - 3, and that the thermophilic microorganisms
adapted to the higher flee-ammonia when the influent concentration was
reduced to 50.4kg VS m -3 and the HRT to 15 days (note that the
thermophilic microorganism had been exposed to the high flee-ammonia
 Anaerobic fermentation of swine manure 187

concentrations for over 170 days when steady state at 15-day HRT
occurred).
Another factor contributing to the instability at 55 °C and 62.5 kg VS
per m 3 was simply the higher influent concentration. We (Hashimoto et
al., 198 la) have reported that CH4 production rates become increasingly
inhibited as the influent VS concentration increases above certain
threshold concentrations. This relationship can be mathematically
described as follows (Chen & Hashimoto, 1978):

7v = ( B o S o / H R T ) ( 1 - K / ( H R T I t m - 1 + K)) (2)

Where Vv= CH4 production rate (m 3 CH 4 per m 3 fermentor per day).

So = tnfluent VS concentration (kg m - 3).
B o = U l t i m a t e CH 4 yield (maCH4 per kg VS fed as HRT
approaches infinity).
HRT = Hydraulic retention time (Days).
/~,~= Maximum specific growth rate (Day-1).
K = Kinetic parameter (Dimensionless).
Equation (2) shows that when S o, B o, HRT and #m are constant, ~'v
decreases when K increases. Figure 5 shows how K increases when the S O
of swine manure increases. The data of Summers & Bousfield (1980) were

,¢

=___r --I

i 210 i i i i ;0
0 I0 30 40 50 60
INFL. V$ CONC. ( K G / m 3 )

Fig. 5. Effect of influent Volatile Solids concentrationon kinetic parameter (K).

188 Andrew G. Hashimoto

used to calculate K, assuming a B o of 0.36 m 3 CH 4 per kg VS fed and a #,,

related to temperature (T in °C) as follows (Hashimoto et al., 1981a):
/~,, = 0.013(T) - 0" 129 (3)
Figure 5 shows that Kis related to S Oby the following empirical equation:
K = 0.50 + 0.0043 exp (0.091So) (4)
Figure 5 and eqn. (4) show that K increases sharply when S O increases
above 50 kg VS per m 3. This may partially explain why the 55 °C fer-
mentors were unstable at 62.5 kg VS per m 3 but stable at 50.4 kg VS per
m 3. The fact that similar instability did not occur at 35 °C, even with the
same K, may be because the free ammonia concentration was significantly
lower at 35°C. Thus, the instability of the fermentation at 55°C and
62-5 kg VS per m 3 was probably due to the combined effects of high free-
ammonia and influent VS concentrations.
The usefulness of eqns (2), (3) and (4) to predict the CH 4 production
rates from swine manure was evaluated by comparing the predicted rates
with the experimental rates. Table 5 shows that Yv for the unstable
fermentation (55°C, 62.5kg VS per m 3, 25-day HRT) was poorly
predicted. This was expected since eqn. (2) was derived assuming stable
steady-state conditions (Chen& Hashimoto, 1978) and, therefore, cannot
be expected to predict unstable fermentations. When the results of the
unstable fermentation were not used, the mean ratio of ~v(predicted) to ~v
(experimental) was 0.99 with a standard deviation of 0.14. The standard
deviation was increased because of the unexpectedly high ~v at 35 °C and
5-day HRT. The reason for this high rate was not known since the
mesophilic fermentor was expected to fail at 5-day HRT. If the result from
that fermentor was not used (since it is inadvisable to operate mesophilic
fermentors at 5-day HRT), the mean ratio of predicted to experimental Yv
was 1.03 with a standard deviation of 0.09. This level of variation is more
than adequate for most fermentor design applications.
The highest ),vachieved in this study was 3.12 m 3 CH 4 per m 3 fermentor
per day at 55 °C, 5-day HRT, S O = 50.4kg VS per m a and loading rate of
10 kg VS per m 3 per day. The CH 4 production and loading rates reported
in this study are ca. 2-5 times higher than the rates reported in the
literature and this shows that significantly higher CH 4 production and
loading rates are possible for swine manure fermentations than previously
thought. With more research and development effort even higher rates
may be possible.
 Anaerobic fermentation of swine manure 189

TABLE 5
Comparison of Experimental and Predicted Methane Production Rates from Swine
Manure

Temperature HRT SO CH 4 production rates Ratio

(°C) (days) (kg VSm -3) (m 3 per m a per day) Pred.~Exp.

Experimental Predicted°

55 25 62-5 0'26 1-08 4.15

55 15 50"4 1'45 1-47 1'01
55 10 50-4 1'80 2-08 1-16
55 5 50"4 3' 12 3-34 1-07
35 25 62'5 1-04 0"98 0"94
35 15 50'4 1-22 1"33 1"09
35 10 50'4 1'91 1-76 0"92
35 5 50'4 2"69 2"01 0-75

a From eqn. (2):

~V= (BoSo/HRT)(1- K/(#mHRT- 1 + K))
where Bo = 0.49m 3 CH4 m 3 fermentor per day; K = 1.77 at So = 62.5kg VS m -3 and
K=0.92 at So=50-4kg VS m -3 (from eqn.(4)) and #m=0.326 per day at 35°C
and #,,= 0'586 per day at 55°C (from eqn.(3)).

ACKNOWLEDGEMENTS

T h e technical a s s i s t a n c e o f L y n n N i e m a n n , Steve S p e l l m a n a n d F r a n
S t o e g e r - W a l l a c e is a p p r e c i a t e d a n d g r a t e f u l l y a c k n o w l e d g e d . R e v i e w a n d
comments by Yud-Ren Chen, Vincent Varel and Michael MacNeil are
also a p p r e c i a t e d .

REFERENCES

American Public Health Association (APHA) (1975). Standard methods f o r the

examination o f water and wastewater (14th edn). American Public Health
Association, Inc., New York, NY.
A O A C (1975). Official methods o f analyses (12th edn). Association of Official
Analytical Chemists, Washington, DC.
Chen, Y. R. & Hashimoto, A. G. (1978). Kinetics of methane fermentation.
Biotechnol. Bioengr. Symp. No. 8, 269-82.
190 Andrew G. Hashimoto

Converse, J. C., Evans, G. W., Robinson, K. L., Gibbons, W. & Gibbons, M.

(1981). Methane production from a large-size on-farm digester for poultry
manure. In: Livestock waste: A renewable resource. Am. Soc. of Agric.
Engr., St.Joseph, MI, 122-5.
CRC (1968). Handbook of chemistry and physics (48th edn). Chemical Rubber
Co., Cleveland, OH.
Fischer, J. R., iannotti, E. L., Porter, J. H. & Garcia, A. (1979). Producing
methane gas from swine manure in a pilot-size digester. Transactions of the
ASAE, 22(2), 370-4.
Fischer, J. R., Sievers, D. M. & Fulhage, C. D. (1979). Design criteria and
operational guidelines for a pilot-scale anaerobic digester. Resource
Recovery and Conservation, 4, l - 11.
Gramms, L. C., Polkowski, L. B. & Witzel, S. A. (1971). Anaerobic digestion of
farm animal wastes. Transactions of thk ASAE, 14, 7-13.
Harvey, W. R. (1975). Least squares analysis of data with unequal subclass
numbers. ARS H-4, US Department I of Agriculture.
Hashimoto, A. G. (198 l). Methane production from beef cattle manure: Effects of
temperature, hydraulic retention time and influent substrate concentration.
Quarterly Progress Report submitted to the Solar Energy Research
Institute, Golden, CO.
Hashimoto, A. G., Chen, Y. R. & Varel, V. H. (1981a). Theoretical aspects of
methane production: State-of-the-art. In: Livestock waste: A renewable
resource. Am. Soc. of Agric. Engr., St. Joseph, MI, 86-91, 95.
Hashimoto, A. G., Varel, V. H. & Chen, Y. R. (1981b). Ultimate methane yield
from beef cattle manure: Effect of temperature, ration constituents,
antibiotics and manure age. Agric. Wastes, 3, 241-56.
Hobson, P. N. & Shaw, B. G. (1976). Inhibition of methane production by
Methanobacterium formicicum. Water Res., 10, 849-52.
Huang, S. Y., Won, K. T. & Wang, H. H. 0978). Optimum design of anaerobic
digester for swine waste. Proc. Nat'l. Sci. Counc. Rep. of China, 2(4), 407-15.
Kroeker, E. J., Schulte, D. D., Sparling, A. B. & Lapp, H. M. (1979). Anaerobic
treatment process stability. J. Water Pollut. Control. Fed., 51, 718-27.
Lapp, H. M., Schulte, D. D., Kroeker, E. J., Sparling, A. B. & Topnik, B. H.
(1975). Start-up of pilot scale swine manure digesters for methane
production. In: Managing livestock wastes. Am. Soc. of Agric. Engr.,
St. Joseph, MI, 234-7, 243.
McCarty, P. L. (1964). Anaerobic waste treatment fundamentals. III. Toxic
materials and their control. Public Works, 95, 91-4.
McCarty, P. L. & McKinney, R. E. (1961). Salt toxicity in anaerobic digestion. J.
Water Pollut. Control Fed., 33, 399-415.
Sievers, D. M. & Brune, D. E. 0978). Carbon/nitrogen ratio and anaerobic
digestion of swine waste. Transactions of the ASAE, 21, 537-41,549.
Stevens, M. A. & Schulte, D. D. (1979). Low temperature anaerobic digestion of
swine manure. J. Env. Engr. Div., ASCE, 105, 33-42.
Summers, R. & Bousfield, S. (1980). A detailed study of piggery-waste anaerobic
digestion. Agric. Wastes, 2, 61-78.
 Anaerobic fermentation of swine manure 191

van Velsen, A. F. M. (1979a). Anaerobic digestion of piggery waste. 2. Start-up

procedure. Neth. J. Agric. Sci., 27, 142-52.
van Velsen, A. F. M. (1979b). Adaptation of methanogenic sludge to high
ammonia-nitrogen concentrations. Water Res., 13, 995-9.
van Velsen, A. F. M., Lettinga, G. & den Ottelander, D. (1979). Anaerobic
digestion of piggery waste. 3. Influence of temperature. Neth. J. Agric. Sci.,
27, 255-67.
Wael, L. L. & Gehrke, C. W. (1975). An automated total protein nitrogen
method. J. Assoc. Official Analytical Chemists, 48, 1221-6.