METHANOGENESIS IN MONOGASTRIC ANIMALS

BENT BORG JENSEN

Danish Institute of Animal Science, Department of Nutrition, Research Centre Foulum, P O. Box 39,
DK-8830 Tjele, Denmark

Abstract. Studies of methanogenic bacteria present in monogastric animals are still scarce. Methanogens have been isolated from faeces of rat, horse, pig, monkey, baboon, rhinoceros, hippopotamus,
giant panda, goose, turkey and chicken. The predominant methanogen in all except the chicken and
turkey is species of Methanobrevibacterium. The chicken and turkey harbour species of Methanogenium. In pig the population of methanogenic bacteria is more than 30 times as dense in the distal colon
as in the caecum. This finding is in agreement with the finding that the rate of methane production
is much higher in the colon than in the ceacum. The amount of methane excreted clearly seems to
depend on the amount of non-starch polysaccharide intake.
The directly measured methane production rate in pigs is from 3.3 to 3.8 times lower than the
amount expected from stoichiometric estimates. These data, together with data showing that only
small net amounts of hydrogen and small amounts of methane are produced in the ceacum and
proximal colon where the microbial activity is high, clearly indicate that hydrogen sinks other than
methane production are involved in hydrogen removal in the hindgut of pigs and probably also in
other monogastric animals.
Methane production by monogastric animals is lower than methane production by ruminants.
However, methane production by large herbivorous monogastric animals such as horses, mules and
asses is substantial (up to 80 1 per animal per day). Methane production by rodents and avians is
low. In general, methane production by wild animals is lower than methane production by domestic
animals. It is concluded that the contribution of monogastric animals to the global methane emission
is negligible, as it only represent about 5% of the total methane emission by domestic and wild
animals of 80 Tg per year.

1. Introduction

Two different types of anaerobic microbial ecosystems produce significant amounts

of methane. One type is the intestinal tract ecosystem such as the complex forestomach of ruminants and the large intestine of monogastric animals. This type of
ecosystem does not completely convert substrates to CH4 and CO2. They accumulate significant quantities of acetate, propionate, and butyrate (Table I). The
predominant substrates for methanogens in these ecosystems are H2 and CO2. The
other type of ecosystem, e.g., swamps, rice paddies, terrestrial and marine aquatic
sediments, and anaerobic digestion systems, are complete bioconversion systems.
Acetate, propionate, butyrate, and H2 and CO2 are formed in the initial stages of
the process. However, propionate and butyrate are further converted to acetate and
H2; acetate, together with Hz and CO2 are substrates for methanogens resulting in
complete bioconversion of organic matter to CH4 and CO2 (Table I).
The principal natural substrates in both types of ecosystem are polysaccharides,
proteins and lipids. Production of acetate, propionate, butyrate, H2 and CO2 requires
Environmental Monitoring and Assessment 42:99-112, 1996.
(~) 1996 Kluwer Academic Publishers. Printed in the Netherlands.

100

BENT BORG JENSEN

Table I
Incomplete and complete anaerobic bioconversion of carbohydrate to methane

Incomplete (gastrointestinal tract)

57.5 C6H1206 --+ 65 CH3COOH + 20 CH3CH2COOH + 15 CH3CH2CH2COOH + 35 CH4 +
60 CO2 + 25 H20
Complete (swamps, ricepaddies, anaerobics sewage digestion systems)
57.5 C6H1206 ~ 172.5 CH4 + 172.5 CO2
Adapted from Miller (1991).

complex integrated activities between several different microbial species. Further,

the removal of H2 by the methanogens has a pivotal influence on the activities of
the non-methanogenic microorganisms in these ecosystems.
A major difference between the two types of ecosystems is turnover time.
Intestinal bioconversion systems have turnover times of approximately 1 to 2 days.
In contrast, the turnover times of complete bioconversion systems are weeks to
months, because microbial conversion of acetate, propionate, and butyrate to CH4
and CO2 involves bacteria that have relative long generation times. These types
of bacteria cannot be sustained in the testinal ecosystem due to their slow rates of
multiplication.
The aim of the present review is to describe the characteristics of the animal intestinal tract ecosystem of monogastric animals with special reference to
methanogenesis.

2. The Digestive Tract of Various Monogastric Animals

There is a widespread tendency to overlook or underestimate the significance of
microbial fermentation in monogastric animals. However, the digestive tract of
some monogastric animals is quite complex and provide many environments for
fermentative bacteria. In general the gastrointestinal tract is a tube extending from
the mouth to the anus. It is divided into various well defined anatomic regions
whose structure and function reflect the diet and the life style of the particular
animal species; as a consequence animal species can be classified into several
groups based on the characteristics of their gastrointestinal tract (Figure 1).
Ruminants represent the most developed and specialized group, from the point
of view of microbial fermentation and the ability to use non-starch polysaccarides
unavailable to animal digestion. However, pregastric fermentation is not restricted
to ruminants, and has been reported in primates, rodents, ungulates and marsupials
(Van Soest, 1984).

METHANOGENESIS1N MONOGASTRICANIMALS

A,) Pony

B) Pig

C) Mink

[3) Goose

101

Figure 1. The gastrointestinal tract of various monogastric animals. (A) Herbivore (horse); (B)
omnivore (pig); (C) carnivore (mink); (D) avian (goose).

102

BENT BORG JENSEN

Monogastric herbivorous show a variety of adaptations. One group includes

large non-ruminant herbivores such as the perissodactyls (horse, rhinoceros, etc.)
and other large herbivores (elephant, etc.). In all of these, microbial fermentation
is more important in the sacculated colon than in the caecum. Herbivorous rodents
and other small herbivorous mammals characteristically exhibit large caeca and
unsacculated colons. Adaptations employing the caecum as the main fermentative
site are often associated with coprophagy as in the rabbit, hare and lemming.
In omnivores such as man and pig the caecum is much reduced, but the colon is
sacculated (Figure 1). The sacculation of the colon and the relative small caecum
may represent a special adaptation of the lower bowl to fermentation.
Carnivores (mink, cat, dog, etc.) comprise another class of mammals. Their
gastrointestinal tract is simple. The intestine is relatively short with little or no
caecal capacity and an unsacculated colon (Figure 1).
The anatomy of the avian gastrointestinal tract is most notably different from
that of mammals in the mouth area, in the presence of a crop in the oesophagus, in
the presence of a muscular stomach or gizzard and in most cases in the presence of
two caeca. The large intestines of birds are short and are not sharply demarcated
from the rectum and small intestine (Figure 1).

3. Microbial Acitvities in Various Regions of the Digestive Tract in Different

Monogastric Animals
The distribution of the microbiota within the gastrointestinal tract differs between
animal species. Investigations of the intestinal bacteria have concentrated on studies
of the human intestine and the rumen. However, our knowledge of the components
of the gut microbiota of the pig and the chicken is slowly but continuously increasing. Information for other species of monogastric animals is much less detailed.
The population level of the microbiota in various parts of the gastrointestinal
tract of monogastric animals depends on the doubling time of the microorganism
under the physico-chemical conditions in the part of the gastrointestinal tract under
investigation, and the emptying rhythm of the particulate part (reservoir). The
stomach and the crop is the first reservoir where the ingesta spend some time,
and where the microflora may multiply. In contrast to humans, pig, rodents and
fowl harbour a permanent microflora in the proximal regions of the digestive tract,
consisting of lactobacilli and streptococci. These permanent populations can be
achieved in the digestive tract of pigs, rodents and fowl because the lactobacilli
and streptococci associate with the stratified squamous epithelial surface lining
that part of the proximal digestive tract of these animal species (Tannock, 1992).
In contrast to the stomach, most of the small intestine is unsuitable for bacterial
proliferation in healthy animals, simply because the intestinal transit is too rapid to
allow time for microbial division. Proliferation occurs in the upper part of the small
intestine only when there is adhesion to the gut wall, or mechanical obstruction

METHANOGENESISIN MONOGASTRICANIMALS

103

Table II
Microbial activityin intestinesfrom variousanimals
Species

Mink
Goose
Duck
Rata
Ratb
Chinchilla
Pig
Pig
Pig
Cow

Body weight
(kg)
ATP
(mg)
1.0
4.0
4.0
0.1
0.1
0.5
10
40
90
500

0.1
0.5
0.6
0.1
0.3
0.5
4.1
27.2
75.9
5000

Intestinal contents
ATP
ATP
(#g (kg BW)-1) (#g (kg BW~
120
130
140
800
2800
1000
420
1030
840
8000

-1)

120
180
200
450
1600
910
750
1800
2600
41000

a Low fiberdiet, b high fiberdiet.

leads to stasis of digesta, with pathological consequences. Alimentary stasis is

the rule of the lower part of the small intestine (ileum) and the large intestine
(caecum, colon and rectum) and all gut bacteria have sufficient time to multiply
there, resulting in a large microbial population. The hydrolytic digestive function
of the large intestine of pigs and other monogastric species is carried out by this
rich and diverse population of anaerobic bacteria (Moore et al., 1987). As shown
in Figure 2, the density of the microbial population in the caecum and colon of pigs
amounts to 101~
bacteria per g of digesta, comprising more than 400 different
species, of which only few have been described in full (Moore et al., 1987).
The microbial activity (measured as the concentration of ATP) in various segments of the digestive tract of several monogastric animals is shown in Figure 3. In
all animal species highest microbial activity was found in caecum and the proximal
segments of the colon; however, it is obvious that microbial activity differs between
the animal species.
Table II shows the total microbial activity found in the entire gut per kg of
metabolic weight of the animal. These data clearly indicate that the microbial
activity is much more important to ruminants than to monogastric animals. Further,
the data illustrate that the microbial activity is higher in pigs than in fowl, and that
the microbial activity depends on the age of the animal and on the diet. Probably the
single most important factor influencing the microbial activity in the gastrointestinal
tract of monogastric animals is the amount and type of substrate available to
the microbiota. In particular, non-starch polysaccarides are the principal energy
substrate for large intestinal microbial fermentation, and the amount as well as the
chemical and structural composition of the carbohydrate are important factors for

104

BENT BORGJENSEN

11~ x ~ X anaerobe
X

10

bacteroides
x ~lactobacilli
~ X

"T
A

~cteria

:oliforme

c~

enterococci

o-1

7
i1
C)
C~

..9.o
6
~ y e a s t
_

Z,
J
;tomach

small intestine

[,..J colon
caecum

Figure 2. The density of selected populations of the microbiota in various regions of the digestive
tract of pigs.

the microbial activity in the digestive tract (Bach Knudsen et al., 1991, 1993a, b;
Jensen, 1988; Jensen and J~rgensen, 1994; Macfarlane and Cummings, 1991).

4. Methanogenic Bacteria in Monogastric Animals

Studies of methanogenic bacteria present in monogastric animals apart from man

is still scarce and mainly qualitative. Methanogens have been isolated from fae-

METHANOGENESIS IN MONOGASTRIC ANIMALS

105

120
rabbit
100

80
c(9

ceecotrofe

C
0

rat
(high fiber)

80
C7~

40
chicken

::1,

20
~ pig
goose

J7, J smelt intestine

stomach
gizzard

]~ colon
caecum

Figure 3. Microbial activity in various regions of the gastrointestinal tract in different monogastric
animals.

ces of rat, horse, pig, monkey, baboon, rhinoceros, hippopotamus, giant panda,
goose, turkey, and chicken (Miller and Wolin, 1986; Miller, 1991). The predominant methanogen in all except the chicken and turkey is species of Methanobrevibacterium (Miller and Wolin, 1986). The chicken and turkey harbour species
of Methanogenium (Miller and Wolin, 1986). Methanobrevibacterium is also the
most common methanogen in faeces of man and insects and in the forestomach of
the bovine rumen (Miller and Wolin, 1986).
Quantification of the number of methanogenic bacteria in the faeces of different
animal species including man has been done by Sorlini etal. (1988). Highest values
were found in pig faeces, followed by human, cattle and rabbit faeces (Table III).
Butine and Leedle (1989) showed that in pigs the population of methanogenic

106

BENT BORG JENSEN

Table III
Numbers of anaerobic bacteria and
methanogenic bacteria present in faeces
from various mammals
Species

Anaerobes a

Methanogens a

Mink
Cattle
Pig

4 x 10 I1
3 x 1011
3 x 1011

1 x 107
1 x 106
1 x 108

Rabbit

7 x 109

4 104

a Bacteria per gram dry weight (MPN).

Data adapted from Sorlini et al. (1988).

bacteria was more than 30 times as dense in the colon as in the caecum; a finding
that is in agreement with the results of Jensen and JCrgensen (1994) who found that
the rate of methane production and the concentration of methane in the caecum
and the proximal colon were low, followed by a steady increase in the successive
segments of the hindgut. This finding is further supported by the results of Robinson
et al. (1989), who found the rates of methane production of colonic samples to be
ninefold higher than those of caecal samples.

5. Methane Production in the Gastrointestinal Tract of Pigs

Apart from man and pig very little information exists about methane production
in the digestive tract of monogastric animals. The main products of non-starch
polysaccharide fermentation in the gastrointestinal tract of pigs are short chain fatty
acids, lactate, and various gases (Macfarlane and Cummings, 1991). The short chain
fatty acids produced are physiologically important especially in the large intestine,
where butyrate in particular is required to maintain the health of the epithelial cells
lining the gut. Three gases, H2, CH4 and CO2, are produced in appreciable volumes
by the intestinal microbiota. The main part is excreted in flatus, while only a smaller
part is absorbed into the bloodstream and excreted in expired air. It has been
indirectly shown by using respirometry that methanogenesis occurs in the lower
gastrointestinal tract of pigs. In contrast to ruminants, methanogenesis accounts
for minimal loss of digestible energy in pigs (Christensen and Thorbek, 1987). A
number of investigators have used breath and flatus H2 and C H 4 measurements to
quantitate fermentation in the large intestine. However, very little information exists
about the relative rates of gas production in different regions of the gastrointestinal
tract of monogastric animals.
In a recent study by Jensen and J~rgensen (1994) the microbial activity, the
composition of the gas phase and the relative rate of gas production were investigated in various regions of the gastrointestinal tract of pigs (120 kg LW) fed either
a low (5% NSP) or a high fibre diet (27% NSP). A dense population of culturable

METHANOGENESIS IN MONOGASTRIC ANIMALS

107

anaerobic bacteria, a high ATP concentration and high adenylate energy charge
(AEC) values were found in the last third of the small intestine, indicating that a
substantial microbial activity takes place in that portion of the gut.
The highest microbial activity (highest bacterial counts, highest ATP concentration, high adenylate energy charge, and low pH) was found in the caecum and
proximal colon. Higher microbial activity was found in the stomach and all segments of the hindgut in the pigs fed the high fibre diet than in the pigs fed the
low fibre diet. Very good correlations were found between the concentration of
gases in specific portions of the gut and the in vitro production rates at the same
site. The highest concentrations and highest production rates for H2 were found
in the last third of the small intestine, while only small concentrations and low
production rates of H2 were detected in caecum and colon. No methane in the
gas phase and no methane production could be detected in the stomach or small
intestine. The production rate and concentration of methane was low in the caecum
and the proximal colon, followed by a steady increase in the successive segments
of the hindgut. A very good correlation was found between in vivo and in vitro
measurements of methane production. The amount of CH4 produced by pigs fed
the low fibre diet was estimated to be 1.4 litres per day per animal (120 kg LW)
and the corresponding amount in vivo to be 1.0 litre per day. Substantially higher
amounts of CH4 were produced by pigs fed the high fibre diet (12.5 litres per day
in vivo and 12.2 litres per day in vitro).
Jensen and JOrgensen (1994) concluded from their results that although the
highest microbial activity was found in the caecum and proximal colon and although
hydrogen production is an obligate part of anaerobic fermentation in the hindgut,
only small net amounts of hydrogen were produced in these segments of the gut.
In contrast to the rumen, where all microbially produced H2 is metabolized to
methane, their results show that only small amounts of methane were produced in
the caecum and proximal colon in pigs. This strongly indicates that hydrogen sinks
other than methane production are involved in hydrogen removal in the caecum
and proximal colon of pigs.
In contrast to man where about half the population are methane-producers, all
pigs seems to produce methane in the hindgut (Christensen and Thorbek, 1987;
Jensen and JCrgensen, 1994; Jcrgensen, unpublished; Zhu etal., 1993). As shown by
Christensen and Thorbek (1987), the daily CH2 excretion by pigs increases linearly
with increasing LW, probably as a result of increasing feed intake. The amount of
CH4 excreted per kg dry matter feed intake varied from 2.5 to 5.2 litres depending
on the composition of the diet. Zhu et al. (1993) and Jensen and J~rgensen (1994)
found that the amount of methane excretion increase with increasing fibre content
in the diet. Calculating the daily amount of non-starch polysaccharide intake in
the experiments of Christensen and Thorbek (1987), Zhu et al. (1993), Jensen and
JOrgensen (1994), and JCrgensen (unpublished results) and plotting this daily NSP
intake versus the daily amount of methane excreted, clearly shows that the amount
of methane excretion by pigs is dependent on the NSP intake (Figure 4).

108

BENT BORGJENSEN

14
0

12
A

"0
C

e ~

0
X

10
O
8

A A

,o

t-

e-

e.6
9
~

2
O
0

100 200 300 400 500 600 700

800 900 1000

NSP consumed (g/day)

Figure 4. Relation between daily amount of non-starch polysaccharides (NSP) consumed and daily

methane excretion in pigs. Symbols correspond to different studies; A, Christensen and Thorbek
(1987); A, Jensen (unpublished); l , Jensen and Jcirgensen (1994); O, Jr
(unpublished); O,
Zhu et al. (1993).

6. Stoichiometric Estimates of Methane Production in Pigs

Methane and propionate are the two major recognized hydrogen sinks in microbial
fermentation in the rumen (Hungate, 1966; Orskov et al., 1968). A stoichiometric
carbon-hydrogen balance equation based on measurements of VFA molar proportions and the amount of carbohydrate fermented can be used to predict the amount
of methane produced (Van Soest, 1982). This technique has been widely used in
ruminant studies and appears to give sensible results (Orskov et al., 1968; Whitelaw
et al., 1970). However, this is not the case for pigs. As shown by Zhu et al. (1993),
the stoichiometric estimates of methane was from 3.3 times (cereal-based diet) to
3.6 times (cereal-based diet supplemented with 30% unmolassed sugar-beet pulp)
higher than those directly measured. The same type of calculations with the data
of Jensen and JCrgensen (1994) and J~rgensen and Jensen (1994) showed that the
directly measured methane production rates were 3.8 times lower than the amount
expected from the stoiochiometric estimate. These data together with the data of

METHANOGENESIS IN MONOGASTRIC ANIMALS

109

Jensen and J~rgensen (1994) showing that only small net amounts of hydrogen
and small amounts of methane were produced in the caecum and proximal colon,
clearly indicate that hydrogen sinks other than methane production are involved in
hydrogen removal in the hindgut of pigs and probably also in other monogastric
animals.

7. Hydrogen Sinks other than Methane Production

As pointed out by Zhu et al. (1993) and Jensen and JCrgensen (1994) a number
of possible pathways for disposal of H2 other than methane production exist in
the gut. These include saturation of unsaturated fatty acids, reduction of nitrate to
ammonia, reduction of CO2 to acetate, reduction of sulphate to sulphide, reduction
of oxygen that diffuses from the blood into the gut lumen and microbial synthesis
of lipid and amino acids.
Unsatured fatty acids are saturated distal to the ileal-caecal junction (J~rgensen
and Just, 1988) and Christensen and Thorbek (1987) have shown that inclusion
of soy-bean oil in a basal diet reduced the amount of methane excreted by pigs.
Further, the pig intestinal microbiota contains acetogenic bacteria (DeGraeve et
al., 1990) and sulphate-reducing bacteria (Butine and Leedle, 1989). It has been
shown that when sulphate is available, sulphate-reducing bacteria have a higher
substrate affinity for hydrogen than methanogenic bacteria and that methane excretion only occurs when sulphate is absent or limiting (Lovley et al., 1982; Lupton
and Zeikus, 1985). On the other hand, it is known that acetogenic bacteria are displaced by methanogenic bacteria in competition for available hydrogen (Prins and
Lankhorst, 1977). Thus acetogenic bacteria will only become active when there
is little hydrogen uptake by sulphate-reducing bacteria or methanogenic bacteria.
However, acetogenesis may be important in the caecum and proximal colon of
pigs, since it has been shown that pH may be an important factor in controlling
the rate of hydrogen uptake (Gibson et al., 1990). In vitro studies have shown
that sulphate-reducing and methanogenic bacteria in human faeces are relatively
pH-sensitive, preferring an environment that is neutral or slightly alkaline, whereas
highest rates of acetogenesis occurred at acidic pH. Since the pH in the caecum
and proximal colon of pigs is low (pH 5.0-5.5) whereas the pH in the distal colon
approaches neutrality (pH 6.5-7.0) acetogenesis may be the dominating hydrogen
sink in the caecum and proximal colon while methanogenesis may dominate in the
distal colon.

8. Methane Production by Various Monogastric Animals

The daily methane production by various domestic and wild monogastric animals
together with the daily methane production by ruminants such as cow and sheep are

110

BENTBORGJENSEN
Table IV
Daily intestinal methaneproductionby variousdomesticand wild animals
Species

Body weight CH4 produced CH4 produced

(kg)
(litre day- t)
(litre day- l
(kg body weight)- 1)

Domestic animals

Cow
Sheep
Horse
Mules/assess
Pig
Man
Goose
Rat

500
40
500
250
100
50
0.4
0.3

230
30
80
40
10
2
0.05
0.001

0.46
0.75
0.16
0.16
0.10
0.05
0.01
0.03

4
20
60
70
110

0.09
0.10
0.07
0.07
0.06

Wild animals

Warthog
45
Zebra
200
Rhinoceros
800
Hippopotamus 1000
Elephant
1700

Data from Champ et al. (1990); Christensenand Thorbek (1987); Crutzen et

(1986); Jensen(unpublished)and Miller (1990).

al.

shown in Table II. It is obvious that methane production by monogastric animals

is lower than methane production by ruminants. However, methane production by
large herbivorous monogastric animals such as horses, mules and asses is also
substantial. Methane production by rodents such as the rat and by avians such as
the goose is low. In general, methane production by wild animals is lower than
methane production by domestic animals, probably due to a lower feed intake.

9. Overall Methane Production by Monogastric Animals

Crutzen et al. (1986) have estimated the global production of methane by several
monogastric domestic animals and have also tried to estimate the contribution of
wild monogastric animals to the global production of CH4. The yearly production
by pigs was estimated as 1.5 kg per pig per year in the developed countries and to
1.0 kg per pig per year in developing countries. Multiplying those values by the
pig population of 500 million in the developing countries and of 300 million in the
developed countries, this yields about 1 Tg CH4 per year.
The mean yearly methane production by horses was estimated to 18 kg per
animal, and the world population of 65 million horses therefore to produce a total

METHANOGENESISIN MONOGASTRICANIMALS

1 11

of 1.2 Tg CH4 per year. A mean production rate of 10 kg per animal per year was
estimated for the global mule and donkey population of 54 million, leading to a
total emission of 0.5 Tg CH4 per year by these animals.
Methane production by wild monogastric animals, of which zebras, elephants
and other large herbivore are the most important contributors, was estimated to be
less than 0.6 Tg per year.
Therefore, as pointed out by Crutzen et al. (1986), it must be concluded that the
contribution of monogastric animals to the global methane emission is negligible,
as it only represents about 5% of the total methane emission by domestic and wild
animals of 80 Tg per year.

References
Bach Knudsen, K. E., Jensen, B. B., Andersen, J. O. and Hansen, I.: 1991, 'Gastrointestinal Implications in Pigs of Wheat and Oat Fractions. 2. Microbial Activity in the Gastrointestinal Tract, Br.
J. Nutr. 65, 233-248.
Bach Knudsen, K. E., Jensen, B. B. and Hansen, I.: 1993a, 'Digestion of Polysaccarides and Other
Major Components in the Small and Large Intestine of Pig Fed Diets Consisting of Oat Fractions
Rich in fl-D-Glucan', Br. J. Nutr. 70, 537-556.
Bach Knudsen, K. E., Jensen, B. B. and Hansen, I.: 1993b, 'Oat Bran but not/3-Glucan-Enriched Oat
Fraction Enhanced Butyrate Production in the Large Intestine of Pigs', J. Nutr. 123, 1235-1247.
Butine, T. J. and Leedle, J. A. Z.: 1989, 'Enumeration of Selected Anaerobic Bacterial Groups in
Caecal and Colonic Contents of Growing-Finishing Pigs', Appl. Environ. Microbiol. 55, 111211116.
Champ, M., Barry, J.-L., Bonnet, C., Berot, S. and Delort-Laval, J.: 1990, 'The Role of Cell Wall
Polysaccharides and c~-Galactosides in the Flatus Induced by the Consumption of a Legume Seed
(Lupin) in the Rat', Sci. Aliments. 10, 317-323.
Christensen, K. and Thorbek, G.: 1987, 'Methane Excretion in the Growing Pig', Br. J. Nutr. 57,
355-361.
Crutzen, P. J., Aselmann, I. and Seiler, W.: 1986, 'Methane Production by Domestic Animals, Wild
Ruminants, other Herbivorous Fauna, and Humans', Tellus 33B, 271-284.
DeGraeve, K. G., Grivet, J. P., Durand, M., Baumatin, P. and Demeyer, D.: 1990, 'NMR Study
of 13CO2 Incorporation into Short-Chain Fatty Acids by Pig Large-Intestinal Flora', J. Gen.
Microbiol. 36, 579-582.
Gibson, G. R., Cummings, J. H., Macfarlane, G. T., Allison, A., Segal, I., Vorster, H. H. and Walker,
A. R. P.: 1990, 'Alternative Pathways for Hydrogen Disposal During Fermentation in the Human
Colon', Gut 31, 679-682.
Hungate, R. E.: 1966, The Rumen and its Microbes, Academic Press, London.
Jensen, B. B.: 1988, 'Effect of Diet Composition and Virginiamycin on Microbial Activity in the
Digestive Tract of Pig', in: Buraczewska, L., Buraczewski, S., Pasturewska, B. and Zebrowska,
T. (eds.), Digestive Physiology of the Pig, Polish Academy of Science, Jablonna, pp. 392--400.
Jensen, B. B. and Jc~rgensen, H.: 1994, 'Effect of Dietary Fibre on Microbial Activity and Microbial
Gas Production in Various Regions of the Gastrointestinal Tract of Pigs', AppL Environ. Microbiol.
60, 1897-1904.
Jr
H. and Jensen, B. B.: 1994, 'The Effect of Dietary Fibre on Digestibility, Microbial
Activity, and Microbial Gas Production in Various Regions of the Gastrointestinal Tract of Pigs',
in: Souffrant, W.-B. and Hagemeister, H. (eds.), Vlth International Symposium on Digestive
Physiology in the Pig, Bad Doberan, Germany, 4-6 October, pp. 273-276.
J~rgensen, H. and Just, A.: 1988, 'Effect of Different Dietary Components on Site of Absorption
Site of Disappearance of Nutrients', in: Buraczewska, L., Buraczewski, S., Pasturewska, B. and

1 12

BENT BORGJENSEN

Zebrowska, T. (eds.), Digestive Physiology of the Pig, Polish Academy of Science, Jablonna, pp.
230-239.
Lovley, D. R., Dwyter, D. E and Klug, M. J.: 1982, 'Kinetic Analysis of Competition Between
Sulphate Reducers and Methanogens for Hydrogen in Sediments', Appl. Environ. Microbiol. 43,
1373-1379.
Lupton, E S. and Zeikus, J. G.: 1985, 'Physiological Basis for Sulphate Dependent H2 Competition
Between Sulfidogens and Methanogens', Curr. MicrobioL 11, 7-12.
Macfarlane, G. T. and Cummings, J. H.: 1991, 'The Colonic Flora, Fermentation, and Large Bowel
Digestive Function', in: Phillips, S. E, Pemberton, J. H. and Shorter, R. G. (eds.), The Large
Intestine: Physiology, Pathophysiology, and Disease, Raven Press, New York, pp. 51-92.
Miller, T. L.: 1991, 'Biogenic Sources of Methane', in: Rogers, J. E. and Whitman, W. B. (eds.),
Microbial Production and Consumption of Greenhouse Gases, Methane, Nitrogen Oxide and
Halomethanes, American Society for Microbiology, Washington D.C., pp. 175-188.
Miller, T. L. and Wolin, M. J.: 1986, 'Methanogens in Human and Animal Intestinal Tracts', System.
Appl. Microbiol. 7, 223-229.
Miller, T. L., Wolin, M. J. and Kusel, E. A.: 1986, 'Isolation and Characterization of Methanogens
from Animal Faeces', System. Appl. Microbiol. 8, 234--238.
Moore, W. E. C., Moore, L. V. H., Cato, E. P., Wilkins, T. D. and Kornegay, E. T.: 1987, 'Effect
of High-Fibre and High-Oil Diets on the Faecal Flora of Swine', Appl. Environ. Microbiol. 53,
1638-1644.
~rskow, E. R., Flatt, W. P. and Moe, P. W.: 1968, 'Fermentation Balance Approach to Estimate Extent
of Fermentation and Efficiency of Volatile Fatty Acid Formation in Ruminants', J. Dairy Sci. 51,
1429-1435.
Prins, R. A. and Lankhorst, A.: 1977, 'Synthesis of Acetate from CO2 in the Caecum of Some
Rodents', FEMS Microbiol. Lett. 1, 255-258.
Robinson, J. A., Smolenski, W. J., Ogilvie, M. L. and Peters, J. P.: 1989, 'In vitro Total-Gas, CH2, H2,
Volatile Fatty Acid and Lactate Kinetics Studies on Luminal Contents from the Small Intestine,
Caecum, and Colon of the Pig', Appl. Environ. Microbiol. 55, 2460-2467.
Sorlini, C., Brusa, T., Ranalli, G. and Ferrai, A.: 1988, 'Quantitative Determination of Methanogenic
Bacteria in Faeces of Different Mammals', Curr. MicrobioL 17, 33-36.
Tannock, G. W.: 1992, 'Genetic Manipulation of Gut Microorganisms', in: Fuller, R. (ed.), Probiotics.
The Scientific Basis, Chapman and Hall, London, pp. 181-207.
Van Soest, P. J.: 1982, Nutritional Ecology of the Ruminant, O & B Books, Inc., Corvallis, Oregon.
Whitelaw, E G., Hyldgaard-Jensen, J., Reid, R. S. and Kay, M. G.: 1970, 'Volatile Fatty Acid
Production in the Rumen of Cattle given an All-Concentrate Diet', Br. J. Nutr. 24, 179-195.
Zhu, J. Q., Fowler, V. R. and Fuller, M. E: 1993, 'Assessment of Fermentation in Growing Pigs Given
Unmolassed Sugar-beet Pulp: a Stoichiometic Approach', Br. J. Nutr. 69, 511-525.