2

The Rumen Fermentation: A Model

for Microbial Interactions in
Anaerobic Ecosystems

MEYER J. WOLIN

1. Introduction

A ruminant can be thought of as a fermentation factory (Fig. 1). The animal

ingests plant polymers (in grasses, hay, corn, silage, etc.) which are the raw
material for its fermentation. Preliminary processing occurs in the oral cavity
and consists mainly of comminution of food by mastication. The plant material
is then swallowed and transported to the ruminant's complex stomach. The
stomach, called the rumen reticulum or, more simply, rumen, is the site of
fermentation. A massive community of microorganisms, bacteria and protozoa,
ferments the plant material to short-chain volatile fatty acids, methane, and
carbon dioxide. The acids are removed from the rumen by absorption into
the bloodstream and are subsequently used as the animal's primary sources of
energy and carbon. Gases are waste products and are removed by belching.
The fermentation provides nutrients and energy for the growth and division
of the microbial populations participating in the fermentation. Microorganisms
and undigested food are semicontinuously removed from the rumen by passage
to the lower part of the ruminant's digestive tract. Digestive processes then
occur that are similar to those of monogastric animals and include gastric diges-
tion in a terminal compartment of the complex stomach called the abomasum

MEYER J. WOLIN. Division of Laboratories and Research, New York State Department
of Health, Albany, New York 12201, U.S.A.

49
so Meyer J. Wolin

Oral Cavity

Grinding of nutrients
(mastication)
Supply of HC0 3 - Venting of CH4 + CO 2
and H20 (salivation (belching)
and drinking)

Rumen

Fermentation
Eh -.4V
pH 6.5
T 39°C
Buffer = HC03- + CO 2

60 mM Acetate Bacteria (10 10 -lO ll /g)

20 mM Propionate Protozoa (10 5 -10 6 /g)
10 mM Butyrate

Energy and biosynthesis Amino acids, fatty

acids and vitamins

Figure 1. Schematic representation of the ruminant as a factory for the conversion of food
to animal products.
The Rumen Fermentation 51

and digestion in the small intestine. Digestion of the microbial mass exiting
from the rumen provides the animal with its major source of amino acids and
water-soluble vitamins. Undigested microbial and feed residues and microor-
ganisms that inhabit the large intestine leave the animal in the feces.
This review will be concerned with the central feature of the factory, the
fermentation in the rumen and the characteristics of the rumen microbial
ecosystem. The environment of the rumen will be described to emphasize the
parameters that are important in its function as a fmely controlled, semicon-
tinuous fermentation apparatus. Major characteristics of the microbial com-
munity will be outlined. Food-chain relations between the rumen microbial
community and the animal and between different microbial populations in the
rumen will be reviewed. A major subject that will be emphasized is how the
populations in the rumen interact to produce its characteristic fermentation
products, the volatile fatty acids, methane, and carbon dioxide. Since the
general characteristics of the rumen are similar to those of other important
microbial ecosystems-e.g., the large intestine of monogastric animals, anaerobic
sediments, and waste fermentation systems-similarities and differences be-
tween these systems will be discussed. Many important and interesting details
of ruminant anatomy, physiology, and nutrition and characteristics of the
rumen microbial community will not be reviewed because the major goal is to
present the general principles that govern the characteristics of the rumen
ecosystem. The reader is referred to several excellent reviews which present
detailed information about the omitted subjects (Bryant, 1959; Hungate et al.,
1964; Hungate, 1966; Janis, 1976; Bryant, 1977). Figure 1 is a flow diagram of
the ruminant factory that can be used as a guide to the following discussion of
the ecosystem.

2. The Rumen Environment

The rumen, as a semicontinuous fermentation apparatus, has several features

that distinguish it from conventional man-made fermentation equipment. The
inner epithelial wall of the rumen is semipermeable and selectively transports
small molecules to and from the blood of the animal. For example, volatile
fatty acids are transported from the rumen to the blood, and bicarbonate ions
are transported in the reverse direction. Neural responses to specific stimuli
activate physiological mechanisms for eructation (to release fermentation
gases), activate contractions to mix and move rumen contents, and activate
the mechanism for regurgitation of solids. The regurgitated solids are reground
in the oral cavity ("chewing the cud") to permit continued comminution of
solid substrates even after they enter the fermentation chamber and fermenta-
tion is initiated. The biological features of the rumen provide elegant mecha-
52 Meyer J. Wolin

nisms for fermentation of solid substrates, product removal, maintenance of

pH, and disposal of fermentation gases.
Although the rumen is a biological fermentation unit, it, like artificial
fermentation units, operates under well· defined environmental conditions
that are extremely important regulators of the kinds, amounts, and physiolog-
ical activities of the microorganisms in the ecosystem. The rumen environment
is anaerobic. The gas phase is essentially all carbon dioxide and methane, al-
though trace amounts of other gases (H2 , N2 , and O2 ) have been reported. The
gas composition is approximately 65% carbon dioxide and 35% methane. Meth-
ane arises strictly as a fermentation product. Carbon dioxide is produced both
from fermentation and by neutralization of acids by bicarbonate ions entering
the rumen from the saliva and blood. The amounts of gases produced from the
fermentation are very large and are sufficient to significantly dilute ingested
gases. A 500-kg bovine eructates approximately 200 liters of methane per
day. Small amounts of air that enter the rumen during swallowing do not, there-
fore, significantly alter the composition of the gas phase. Any small amounts
of O2 entering the system are quickly used up by oxidative metabolism by
rumen microorganisms. This oxidative activity represents only a minute portion
of the metabolism of the tremendous amount of organic matter entering the
ecosystem. In this sense, the rumen is like a highly eutrophic aquatic environ-
ment where limiting O2 can support the oxidation of only an insignificantly
small amount of the organic matter in the system. The oxidation-reduction
potential of the rumen is low and is approximately -350 mV (E~ at pH 6.5),
which is close to the oxidation-reduction potential of the hydrogen electrode
at the same pH.
The pH of the rumen is approximately 6.5. Neutral plant polymers are
fermented to predominantly acid products, and the pH would drop significantly
during fermentation if it were not for the strong bicarbonate-carbon dioxide
buffering system of the rumen. Salivary bicarbonate is the major neutralizing
agent for the acids produced during fermentation. The temperature of the
rumen is maintained at approximately 39°C through physiological regulation
by the animal.
It is important to emphasize the semicontinuous nature of the fermentation
system. A 500·kg bovine has approximately 70 liters of contents in its rumen.
During a single day, approximately 100 liters of liqUid enters the fermentation
vat (about 60 liters of saliva and about 40 liters of drinking water). In order to
maintain a volume of 70 liters, 100 liters has to be removed from the rumen
each day. This would be equivalent to a dilution rate of 1.4 per day (or a turn·
over time of 0.7 days) for a truly continuous culture. The system, however, is
not truly continuous. Feeding of nutrients is sporadic, as is the inflow of liqUid,
and the nutrient suspension is not homogeneous. In addition, mixing in the
The Rwnen Fermentation 53

rumen is imperfect, and the regurgitation of ingested material adds to the dis-
continuity of the system. Solids have a turnover time that is approximately three
times longer than liquids.
The environmental parameters set some obvious constraints on the popula-
tions of microorganisms one might expect to fmd in the ecosystem. Obligate
aerobes would not be expected to occur in significant concentrations. Dominant
species have to be either facultative anaerobes or anaerobes. The dilution rate
sets some rough upper limits on the growth rate constants or generation times of
organisms that can be maintained in the ecosystem. Some of the microorganisms
are associated with solids, and it can be roughly estimated that associated orga-
nisms with generation times of about two days or less would not be washed out
of the ecosystem. The protozoa of the rumen have generation times of the order
of a day or two and probably would not be able to remain in the rumen if it was
a homogeneously mixed reactor with a turnover time of 0.7 days. Most of the
bacteria in the system have much shorter generation times than 0.7 days. The
methane-forming bacteria do not, and they remain in the system because the
dilution rate can accommodate their long generation times. On the other hand,
much longer turnover times, i.e., five to ten days, would probably encourage the
development of bacteria that can participate in the complete bioconversion of
organic carbon to methane and carbon dioxide as occurs in waste disposal sys-
tems. This would be detrimental to the animal because it would no longer have
the volatile fatty acids available that it needs as sources of carbon and energy in
its metabolism. The elemental carbon, hydrogen, and oxygen of the acids would
end up instead as carbon, hydrogen, and oxygen of methane and carbon dioxide.
Temperature, oxidation-reduction potential, and pH are constraints on the
types of microorganisms that can exist in the rumen only in the sense that
microorganisms would not be present that have obligatory requirements for
conditions that are significantly different from those found in the rumen. It is
not unreasonable to argue, however, that evolution of the system has selected
against organisms that have ranges of activity that include but go far beyond the
normal environmental ranges of the ecosystem. For example, O2 -using facultative
anaerobes have genetic information and enzymes for metabolic activities they
would never use in the rumen environment. Their presence in significant concen-
trations would have been selected against in favor of organisms that do not have
any significant ability to use O2 • Similarly, it would be expected that evolution
has selected for organisms with optimal growth temperatures close to that of the
environment and with maximal and minimal temperatures not far from the
temperature of the rumen. In general, the physiological characteristics of micro-
organisms reflect the evolution of the most efficient physiological mechanisms
for growth and survival in the natural habitat.
The rumen environment is described in detail in Hungate (l966).
S4 Meyer J. Wolin

3. Rumen Bacteria and Protozoa

Almost all the significant rumen bacteria are nonsporing anaerobes. A few
species of facultative anaerobes are present that have no significant ability to use
O2 • Occasionally, significant concentrations of spore-forming anaerobes have
been detected in the rumen.
A few words are necessary to clarify what constitutes a significant bacterial
species (Bryant, 1959). The rumen is an open system. Any microorganisms that
enters or exists in the oral cavity can enter the rumen. Most will not grow or will
grow very poorly in the rumen, but as long as they are not killed they can be
isolated from the rumen by selective procedures. Criteria have to be established
to separate transients and those that barely manage to eke out an existence in
the community from the dominant, important species. Since total counts of
bacteria are about 1 X 1010 to 5 X 1010 per gram of rumen contents, it is com-
mon practice to consider species whose concentrations are 10 8 _10 10 per gram as
members of the dominant community. It has always been the case that these
species demonstrate physiological and nutritional characteristics that are consis-
tent with the characteristics of the ecosystem. There is always the possiblity of
neglecting large sized, less numerous species which may significantly contribute
to bacterial mass and activity, or organisms in low concentration which may have
high activity for some critical ecosystem function. Practically speaking, however,
it has been possible to associate most of the important rumen activities with
species that are present in high concentrations. Genera and species of protozoa
can be identified and enumerated by microscopy. This provides a more direct
method for evaluating the significance of specific taxa than is possible with most
of the bacteria.
Table I shows approximations of the mass of bacteria and protozoa in the
rumen and estimates of the amount of protein they contribute to rumen con-
tents. Rumen contents represent approximately 15% of the live weight of a
bovine and contain about 65% of the weight of food ingested by the animal.
Table II shows the distribution of material in the intestinal tract of a cow as
percentages of body weight and daily intake of food. Rumen contents are ap-
proximately 10-18% dry matter (Hungate, 1975). Microbial cells constitute
about 5% of the dry matter, and they represent about 1% of the dry weight of
the entire animal (Hungate, 1966; Bryant, 1977). The microbial mass is usually
equally distributed between bacteria and protozoa (Table I). Although rumi-
nants can be reared without protozoa by the use of various methods to defaunate
or to prevent the development of the fauna (Becker, 1932; Eadie, 1962), the
typical rumen is a faunated system. Protozoa are important to, although not es-
sential for, the function of the rumen. Because metabolic activity is significantly
greater per cell as cell volume decreases, the contribution of bacteria to the over-
all fermentation in the rumen is Significantly greater than that of the protozoa.
The Rumen Fermentation ss

Table I. Dry Weight (g) of Bacteria and Protozoa and

Microbial Protein in the Rumena

Ruminant

Microorganism Bovine b Ovinec

Protozoa
Cells 315 22.5
Cell protein 172 12.3
Bacteria
Cells 399 28.5
Cell protein 217 15.5

aValues are based on data in Hungate et al. (1971). Total bacteria were 0.57 g (dry weight)/
100 ml (3.1 X 10 10 cells/mI). Total protozoa were 0.45 g (dry weight)/ 100 ml (2.4 X lOs
cells/mI). Protein is estimated as 54.5% of the dry weight (Reichl and Baldwin, 1975).
bBased on 70 liters of rumen contents.
C Based on 5 liters of rumen contents.

Most of the important species of rumen bacteria are listed in Table III with
their major fermentation products and their major energy sources. The impor-
tant cellulolytic species are Ruminococcus albus, R. fiavefaciens, Bacteroides
succinogenes, and possibly Butyrivibrio fibrisolvens. Important starch-digesting
species include B. fibrisolvens, Selenomonas ruminantium, Bacteroides amylo-
philus, Streptococcus bovis, and Succinomonas amylolytica.
The protozoa of the rumen are anaerobic ciliates. There are two major
groups, the holotrichs and the entodiniomorphs (also called oligotrichs). The
holotrichs have cilia all over the cell surface. They are represented by the genera
Dasytricha and Isotricha. Both ferment soluble sugars, and Isotricha ferments
starch. Entodiniomorphs have their cilia arranged as complex organelles. One
genus, Diplodinium, ferments cellulose and starch, and other genera ferment
starch, hemicellulose, and pectin. The entodiniomorphs have only a very limited

Table II. Contents of Digestive Organs of a Milk Cowa

Wet wt. Dry wt.

Organ kg % wt. of animal· kg % Daily food intake

Rumen reticulum 81.8 15.4 8.56 64.8
Omasum 11.0 2.1 0.70 5.3
Abomasum 3.6 0.7 0.27 2.0
Small intestine 9.4 1.8 0.27 2.0
Large intestine 8.6 1.6 0.38 2.9

a From Hungate (1966). The weight of the cow was 533 kg, and the daily food intake was
13.2 kg (dry wt.) of hay.
 \I>
0\

Table III. Fermentation Products and Energy Sources of Important Rumen Bacteriaa

Energy sourcesc
Major fermentation
products b Species Cellulose Starch Xylan Glucose Lactate Glycerol

A, S Bacteroides succinogenes + :;: +

A,S,F B. amylophilus +
A,S,F B. ruminicola ± ± +
A,S,F,H Ruminococcus flavefaciens ± ± ±
A,S Succinivibrio dextrinosolvens +
S Succinomonas amylolytica + +
A,S Treponema sp. +
A,E,F,H Ruminococcus albus ± ± :;:
A,E,F,L,H Lachnospira mUltiparus :;: +
B,F,H Butyrivibrio fibrisolvens :;: ± ± +
B,F,L Eubacterium ruminantium ± +
A,P,B,H Megasphaera elsdenii + + ±
A,P,L Selenomonas ruminantium ± + :;: :;:
A,P,H Veillonella alcalescens +
L Streptococcus bovis + +
L Lactobacillus vitulinus +
CH 4 Methanobacterium
ruminantium H2 and CO 2, formate

a Adapted from Bryant (1977). a::

b Abbreviations: A, acetate; S, succinate; F, formate; H, hydrogen; E, ethanol; B, butyrate; P, propionate; L, lactate. CO 2 is produced by many and ~
used by a few species during fermentation. 2
c+, Used by all strains; ±, used by most strains; :;:, used by few strains; -, not used by any strains. !-'
~
~
The Rumen Fennentation 57

ability to use soluble carbohydrates. Major products of the protozoan fermenta-

tion are acetate, butyrate, lactate, H2 , and carbon dioxide. Propionate may be a
minor product. Formation of ethanol and succinate has not been reported.
Detailed information about rumen bacteria and protozoa can be found in
Bryant (1959), Hungate et al. (1964), and Hungate (1966).

4. Interactions between Microorganisms and the Animal

The ruminant provides nutrients that support the growth of rumen micro-
organisms. All of the necessary carbon, nitrogen, phosphorus, sulfur, and trace
elements are present in the animal's food. The ruminant also contributes sub-
stantially to the maintenance of appropriate physical and chemical conditions
for its fermentation factory; e.g., it contributes to temperature and pH control,
and it controls the dynamics of turnover of the contents of the rumen.
In return for the provision of this excellent habitat, rumen microorganisms
provide activities and products that are essential for the animal. First and fore-
most of these are the microbial cellulases. Cellulose is the most important source
of carbon and energy in the ruminant's diet, but the animal itself does not
produce enzymes for hydrolyzing cellulose. As the microorganisms use cellulose
and other plant carbohydrates as their sources of carbon and energy, they
produce large amounts of acetic, propionic, and butyric acids which the animal
uses as its sources of carbon and energy. Table IV shows estimates of the amounts
of these acid products produced per day in the ovine and bovine. Very large

Table IV. Daily Production of Volatile Fatty Acids

in the Rumena

Ruminant

Volatile acid Bovine b Ovinec

Acetic
mol per rumen 62 4.4
kg per rumen 3.7 0.26
Propionic
mol per rumen 15 1.1
kg per rumen 1.1 0.08
Butyric
mol per rumen 7 0.5
kg per rumen 0.6 0.04

aValues are estimates based on rates of production of 3.7,

0.9, and 0.4 mmol/100 g per hr for acetic, propionic, and
butyric acid, respectively (Hungate et al., 1971).
bBased on 70 kg of rumen contents.
C Based on 5 kg of rumen contents.
58 Meyer J. Wolin

amounts become available to the animal from the microbial digestion of plant
matter.
It is important to recognize that almost all organic plant constituents that
enter the rumen will undergo microbial fermentation whether or not the animal
itself has the enzymatic ability to digest them. For example, the ruminant does
produce pancreatic amylase which is secreted into the small intestine, but the
only starch available to it is that which escapes microbial digestion and the
starch·like polysaccharides stored within microbial cells. Proteins entering the
rumen are also rapidly hydrolyzed and fermented. The animal's proteases are
used mainly to hydrolyze the microbial protein of the rumen after it enters the
stomach·like portion of the rumen, the abomasum, and the small intestine.
Discussions of the interrelationships between the microbial community and
the animal can be confusing if the ruminants being considered are beef cattle fed
using modern methods for the rapid production of tender beef. These animals
are often fed large amounts of starch rather than cellulose. If starch and high·
quality proteins are available for feeding, the husbandryman is often frustrated
by the fact that whatever is fed ends up as whatever the microorganisms produce.
There are certain economic scenarios where it is easy to imagine that it would be
more beneficial to bypass the microbial community with feeds the animal can
directly metabolize because the feed ingredients contain more energy and higher-
quality protein than the microbial products. These are, however, very recent and
very practical scenarios. One only has to consider a cow grazing in a pasture or a
mountain goat nibbling on brush to realize that the rumen system evolved to
permit the utilization of plants that many animals cannot digest. Celluloses and
hemicelluloses are the main sources of carbon and energy for the ruminants
except for the specialized feeding now used to prepare beef cattle for market.
The animal generally benefits from the upgrading of protein quality that occurs
when poor-quality plant proteins are fermented in the rumen and microbial
protein is synthesized and is the major protein subjected to intestinal digestion
(Smith, 1975). This upgrading of quality is actually taken advantage of in modern
feeding where it is sometimes economically advantageous to feed chemically
synthesized urea. The urea is rapidly broken down to ammonia and carbon
dioxide by rumen microorganisms, and the ammonia is used as a source of nitro-
gen for the synthesis of microbial protein (Smith, 1975). A ruminant, therefore,
can be maintained on a diet that contains a significant amount of the animal's
nitrogen requirement in the form of urea.
In addition to providing protein and a method for using cellulose and hemi-
cellulose, the microbial community synthesizes all of the B vitamins required by
the animal. These are released when the microorganisms are digested by the
animal. The vitamins are also in high concentration in the fluid of the rumen.
Ruminants can suffer indirect B-vitamin deficiencies if vitamin synthesis does
not take place in the rumen. When animals are reared on crops raised on cobalt-
The Rumen Fermentation 59

deficient soils, there is insufficient cobalt for microbial synthesis of vitamin B12,
and the animal suffers a vitamin-B12-deficiency disease (Hungate, 1966).
Long-chain fatty acids of microbial lipids are also used by the animal intesti-
nal digestion of microbial cells. The fatty acids are directly incorporated into the
lipids of the animal. The origin of the fatty acids of the ruminant's lipid can be
identified because of the unique structures of the microbial fatty acids. Some
microbial populations also reduce dietary unsaturated to saturated fatty acids,
which are then incorporated into ruminant fats.
The interrelationships discussed above probably apply to many animals other
than domesticated ruminants that have digestive systems that include a fore-
stomach fermentation of plant matter. Some examples are the camel, the kan-
garoo, the hippopotamus, and a few species of herbivorous monkeys (Bauchop,
1971; Janis, 1976). The microbial communities of these animals have not been
studied in detail, but there is good reason to believe that the general principles
governing the operation of these ecosystems are similar to those of domestic
ruminants. There are also many herbivores which use a typical monogastric
digestion system prior to the digestion of celluloses and hemicelluloses in a por-
tion of the large intestine (e.g., rodents and horses). These herbivores use primar-
ily microbial fermentation for digestion of cellulose and hemicellulose in the
posterior portion of the digestive system. Short-chain volatile fatty acids and
methane are formed. Although it would be expected that there will be some
similarities between the large-intestine fermentation systems and the rumen, it
would also be expected that there will be many differences because of the dif-
ference in the dynamics of nutrient passage, the more solid nature of contents
of the large intestine, and the lack of salivary ingredients. Unfortunately, the
large-intestine ecosystems have not been studied to an extent that permits exten-
sive comparisons with the rumen system.
Additional information about the interactions between rumen microorga-
nisms and the animal can be found in Hungate (1966).

5. Nutritional Interactions between Microbial Populations

Nutritional interrelationships between animal and microorganims are of

fundamental importance to the maintenance of the ecosystem. There are also
important interrelationships between the microbial populations of the rumen
which are essential for maintaining the indigenous flora and fauna. There are
classical food-chain relationships where the product of digestion by one species
serves as a nutrient for another species. Food chains involving carbon and energy
sources will be discussed below when fermentation interactions are considered.
Nitrogen utilization in the rumen is to some extent related to fermentation but
will be discussed here because fermentation of nitrogen compounds produces
60 Meyer J. Wolin

only a small contribution to products as compared to the fermentation of

carbohydrates.
The metabolism of protein in the rumen is summarized in Fig. 2. Proteins
entering the rumen are digested and fermented with the release of nitrogen as
ammonia (Smith, 1975). Fermentation is rapid and complete. Amino acids do
not accumulate in significant concentrations in the environment (Wright and
Hungate, 1967). Consequently, the most available form of soluble nitrogen for
microbial biosynthesis is ammonia rather than any form of organic nitrogen.
Almost all of the major species of rumen bacteria can use ammonia as the major
precursor of the cellular nitrogen compounds (Bryant and Robinson, 1962;
Bryant, 1974). In fact, several important species use only ammonia and are in-
capable of growth with amino acids, presumably because they are impermeable
to amino acids (Allison, 1969).
Bacteroides ruminicola uses either peptides or ammonia but not amino acids
as a source of nitrogen (pittman et al., 1967). The peptides have to contain
between 4 and 20 amino acids. Experiments with rumen contents indicate that
proteins are hydrolyzed to peptides that are small enough to enter bacterial
cells, where they are used as nitrogen sources and also catabolized to yield am-
monia without accumulation of consequential amounts of extracellular amino
acids (Wright, 1967). The nitrogen and carbon of amino acids added to rumen
contents are catabolized and are inefficiently incorporated into bacterial protein
(Wright, 1967). The ability of some of the major species of rumen bacteria to
use ammonia and amino acids as major sources of nitrogen for growth is sum-
marized in Table V.

ProtozoaI protein
--+
Bocterial protein

j stro ight - cha in

Other C sources

VFA. aromatic ~C02

aCidj.
. . ._............---..... Urea
acids

....... Major impor ta nce

~ Moderate irTIportonce
c=:::::::::> Minor importance

Figure 2. Metabolism of protein in the rumen. VF A, VoIatile fatty acids.

The Rumen Fermentation 61

Table V. Use of Ammonia and Amino Acids as Major Sonrces of

Nitrogen by Rumen Bacteria

Nitrogen sourceQ

Species Ammonia Amino acids

Bacteroides succinogenes +
B. amylophilus +
B. ruminicola +
Ruminococcus flavefaciens +
R. albus +
Butyrivibrio ftbrisolvens + +
Megasphaera elsdenii +
Selenomonas ruminantium + +
Streptococcus bovis + +
Methanobacterium ruminantium +
Q+, Used by all or most strains; -, not used.

Another habitat-oriented nutritional requirement of at least some rumen

bacteria is associated with the catabolism of proteins and amino acids. The car-
bon skeletons of the amino acids are fermented to derivative acids, fatty acids
from aliphatic amino acids and aromatic acids from aromatic amino acids. (el-
Shazly, 1952; Scott et al., 1964). The products of fermentation of branched-
chain amino acids are branched-chain fatty acids with one carbon less than the
parent amino acids. Isobutyric acid is derived from Val, isovaleric acid from TIe,
and 2-methylbutyric acid from Leu. Several bacterial populations have nutri-
tional requirements for the branched-chain fatty acids (Bryant, 1974). In some
cases, they are used by the organisms for the synthesis of branched-chain amino
acids (Allison, 1969). This is accomplished by reductive carboxylation to pro-
duce the keto-acid homologue of the amino acid followed by introduction of the
amino group. Thus, isobutyric acid is reductively carboxylated to a-ketoiso-
valeric acid with subsequent amination to produce Val. Leu is produced from
2-methylbutyric acid and TIe from isovaleric acid by similar reactions. Some
populations use the branched-chain fatty acids as precursors for the synthesis of
branched long-chain fatty acids which are incorporated into cellular lipids
(Bryant, 1974).
Rumen protozoa do not have the biosynthetic capacity to synthesize amino
acids. The bacteria of the rumen are very important to the protozoa as sources
of nitrogen. Many of the protozoa, particularly the entodiniomorphs, ingest
bacteria, digest their proteins to amino acids, and resynthesize protozoan protein
from the amino acids (Coleman, 1975). Some of the protozoa are able to use
free amino acids as sources of nitrogen (Coleman and Laurie, 1977).
Although the ruminant does not require B vitamins in its diet, most of the
62 Meyer J. Wolin

major bacterial species in the rumen require a few B vitamins when grown in
defmed media. This indicates that some populations of rumen bacteria produce
the B vitamins that others require. Two unusual growth factor requirements that
apply only to bacterial interactions are requirements for heme and for coenzyme
M. Heme is required for growth by some populations and is produced by others
(Caldwell et ai., 1965; Reddy and Bryant, 1977). Requirements for heme for
growth of human large-intestine anaerobes are also common (Sperry et ai.,
1977). Coenzyme M appears to be unique to methanngens and to their metab-
olism. It is produced by some methanogens and used by others (Taylor et ai.,
1974).
The nutritional requirements of rumen microorganisms are, therefore, re-
lated to the nutrients available to them in the more or less steady-state environ-
ment of the rumen. The composition of the food eaten by the animal gives very
little indication of what individual species of rumen microorganisms require for
growth. Many of the nutritional requirements are unique to the rumen or at least
to it and similar habitats. In addition to the requirements mentioned previously,
several rumen species have absolute requirements for sodium (Caldwell and
Hudson, 1974), which is in high concentration in the rumen. Large amounts of
sodium are present in ruminant saliva. Many populations in the rumen require
carbon dioxide for growth (Dehority, 1971). Its prominence in the rumen has
already been mentioned. It is of little use to cultivate rumen microorganisms
on so-called rich media, e.g., blood, milk, or many of the complex commercial
media that contain a variety of protein digests and yeast, liver, or soybean ex-
tracts. Although these media may contain some of the ingredients required by
some of the major rumen populations, they are generally deficient in some of
the important ingredients these organisms have available to them in the rumen.
There is really nothing novel about the relationship between an organism's nutri-
tional requirements and the composition of its natural habitat. It has long been a
practice in microbiology to attempt to use habitat-simulating media to isolate
organisms from a habitat, e.g., milk for lactic acid bacteria and blood for blood
pathogens. The special feature of habitats that contain a complex microbial
community is that the nutrient composition of the steady-state habitat greatly
differs from the composition of the nutrients used to feed the system.

6. Fermentation futeractions

6.1. Food Chains

There are a few examples where products of the energy metabolism of one
rumen microbial population serve as the sources of energy for other populations.
The most prominent example is the chain that leads to the formation of meth-
The Rumen Fermentation 63

ane. Rumen methanogenic bacteria use either hydrogen and carbon dioxide or
formate as substrates for the production of methane (Hungate, 1967; Hungate
et al., 1970). The reduction of carbon dioxide to methane and the essentially
equivalent production of methane from formate provide energy for the growth
of the rumen methanogens. Methanogens from other environments produce
methane from acetate or methanol (Mah et al., 1977; Zeikus, 1977), but these
are not substrates for ruman methanogenesis. There are no methanogens known
that use carbohydrates, amino acids, purines or pyrimidines, or any substrates
other than those cited above as sources of energy. Carbohydrate-fermenting
bacteria in the rumen produce the hydrogen and carbon dioxide and formate
that the rumen methanogens use for the production of methane. Hydrogen and
carbon dioxide are by far the major precursors of methane in the rumen (Hun-
gate et al., 1970).
Another food chain that is of occasional importance is the production of
lactic acid by some rumen bacteria and its use by others. Ordinarily, lactate is
not an intermediate in the overall fermentation. It is produced and used when
animals are fed considerable quantities of easily digestible, starch-containing
diets. Starch-fermenting species such as S. bovis and S. ruminantium are capable
of producing considerable amounts of lactate. Lactate-using species, such as
Megasphaera elsdenii and Veillonella alcalescens, are able to use lactate as an
energy source. There are also lactate-fermenting strains of S. ruminantium
present in the ecosystem.

6.2. Products of Polymer Hydrolysis

It is reasonable to emphasize the ability of organisms to directly use nutri-

ents that enter the ecosystem as sources of energy. Most of the rumen bacteria
and protozoa can use one or another of the plant carbohydrates ingested by the
animal (Table III). There are some carbohydrate-fermenting species and strains
of species that do not attack cellulose, starch, or hemicellulose (Table III).
Since these organisms are present in high concentrations in the environment, it
is likely that they feed on soluble carbohydrates produced by starch- and cellulose-
fermenting populations. This of course assumes that they are not using other
organisms' fermentation products or unidentified energy sources. Cross-feeding
at the level of the soluble products of polymer hydrolysis is a poorly understood
facet of the interaction between populations in the rumen and in other complex
microbial ecosystems. A few general points will be discussed here to indicate the
possible significance of these interactions and some directions to be taken in
future research to clarify their nature and significance.
An obvious, fundamentally important, but often glossed-over feature of the
rumen and other complex microbial ecosystems is that most of the important
carbon and nitrogen sources are introduced as insoluble polymers. Extracellular
64 Meyer J. Wolin

enzymes have to be used to digest these polymers. Products are formed that
enter the cells of the organisms that digest the polymers, but the products are
also available to other species that do not participate in the primary events of
hydrolysis. Competition can take place for the penneating monomeric or oligo-
meric prodUcts. Competition is not necessarily restricted to the organisms that
digest polymers and those that do not. Competition for products can occur
between polymer-attacking species. Obviously, competition cannot operate to
the detriment of populations that have the primary tasks of initiating the diges-
tion of high-rnolecular-weight nutrients. The community would not be able to
survive such extreme competition. Sufficient cross-feeding can occur, however,
to maintain species that cannot ferment polymers per se and to influence the
dynamics of the digestion process. Important considerations in this competition
are the nature of the soluble products of hydrolysis and the permeability of
organisms in the ecosystem to these products.
An example of this cross-feeding is provided by the demonstration of the
growth of cocu1tures of S. ruminantium and B. succinogenes on cellulose. B.
succinogenes is cellulolytic, and S. ruminatium is not. When the two specieS
are grown together with cellulose as the sole energy source, the selenomonad
grows as well as the cellulolytic species {Scheifmger and Wolin, 1973}. The
nature of the soluble products of cellulose hydrolysis preferred by each organism
is not kn()wn. Both species grow well together when cocultured on cellobiose or
Glc, which both can use independently.
Cross-feeding of products from nonpermeating substrates occurs with sub-
stances other than carbohydrates. Lipids are hydrolyzed in the rumen to yield
glycerol, which is fermented by glycerol-using organisms. The relationships
between products of protein hydrolysis and nitrogen requirements were discusseq
previously. Cross-feeding of products of protein hydrolysis is no doubt impor-
tant in protein metabolism, but specific intermediates have not been identified.
Further investigations are necessary to identify the permeating products of the
breakdown of all substrates. It is the permeability of dominant species to specific
products of hydrolysis that is important in many microbial ecosystems.

6.3. Decarboxylation of Succinate

Figure 3 depicts a scheme for the overall fermentation of plant polysac-

charides by the rumen microbial community. Portions of the scheme have
already been discussed. Cellulose and starch are hydrolyzed to permeable
products. These are fermented by the hydrolyzing populations and populations
that feed on the hydrolysis prodUCts. Fermentation produces the end products
of the rumen fermentation, and major and minor extracellular intermediates.
The minor intermediates include lactate and formate; lactate is only occasionally
formed on specific diets, and formate does not appear to be formed in large
The Rumen Fermentation 6S

ACETATE BUTYRATE PROPIONATE C02 CH4

t-
SUC\,NATE
C02
r-
tMethanogens

j2
C02

~C02
I
Lactate fermenters ----- - - - -- _FORMATE
,----~-----~--
'-LACTATE----- - ---

Polymer hydrolyzers; other sugar

fermenters
SOLUBLE SUGARS

tPolymer hydrolyzers
CELLULOSE. STARCH

Figure 3. Intermediates and products of the rumen fermentation of plant polysaccharides;

the dashed line represents minor pathways.

amounts. Lactate, when formed, is fermented to the same products formed from
carbohydrates. Formate is converted to hydrogen and carbon dioxide, which
are then converted to methane. Hydrogen and carbon dioxide from formate
and from other, more important intracellular intermediates are important
precursors of methane. Succinate is an important extracellular intermediate in
the overall rumen fermentation. It is decarboxylated essentially as fast as it is
formed (Blackburn and Hungate, 1963). The extracellular pool of succinate is
extremely small.
Monocultures of many important bacterial species of the ecosystem produce
succinate as a major product of carbohydrate fermentation. Very few species
produce propionate. The major propionate producers in the bovine rumen are
S. rnminantium and M elsdenii. The former species forms propionate from
carbohydrate by a pathway similar to that used by the propionibacteria, the
randomizing pathway. Succinate is an interme;diate (intracellular) of the ran-
domizing pathway. It can be decarboxylated by a series of enzymes to propionate
and carbon dioxide. S. rnminantium can decarboxylate succinate to propionate
and carbon dioxide. Experiments with cocultures of B. succinogenes or R.
flavefaciens and S. rnminantium demonstrate that the selenomonad is able to
grow on carbohydrates produced from cellulose by either cellulolytic species.
The selenomonad produces propionate from carbohydrate and decarboxylates
the succinate produced by the other species to propionate. The net result is
a coculture fermentation that produces propionate instead of succinate. The
interaction is depicted in Fig. 4. The selenomonad obtains no apparent benefit
from carrying out the decarboxylation of succinate produced by the succinate-
66 Meyer J. Wolin

j
CEllULOSE

Cellulolytic species

CELLULOSE FRAGMENTS

Succinate- Propionate-
producers producers
Succinate
pathway

OTHER PRODUCTS Acetate

+ + Figure 4. Interaction between species in-
~S~U~C~C:!:IN~A~T~E~------I"-P r 0 p ~nat e volved in the production of succinate and its
decarboxylation to propionate in the rumen.
CO 2 From Wolin (1974); reprinted by permission.

producing species of the rumen. It is not clear why this two-species system has
developed for the production of propionate in the rumen.
M. elsdenii does not use the randomizing pathway for production of pro-
pionate and presumably does not decarboxylate succinate to propionate. V.
alcalescens uses the randomizing pathway and does decarboxylate succinate.
Although found in the rumen of sheep, Veillonella strains have not been shown
to be important in cattle. Because Veillonella strains do not use carbohydrates
or glycerol but are able to ferment lactate, they are probably important in sheep
only when the diet leads to conditions for the production of significant amounts
of extracellular lactate.
Recent experiments suggest that the interactions necessary for propionate
formation in the rumen may not occur in the human intestinal tract. Many of
the dominant nonsporing anaerobes, particularly Bacteroides, produce succinate
as a major product of carbohydrate fermentation in monocultures. When supple-
mented with vitamin B12 , however, several of these same species produce
propionate instead of succinate, or at least less succinate and more propionate
(Table VI). One of the important enzymes in the formation of propionate from
succinate is methylmalonyl-coenzyme A isomerase, which requires a coenzyme
form of vitamin B12 for activity. Human intestinal Bacteroides apparently can
synthesize the apoenzyme but cannot synthesize the vitamin B12 necessary for
synthesis of the coenzyme. Conversion of succinate to propionate in the large
intestine is probably a single-organism, intracellular process that depends on the
presence of vitamin B12 • The reason for the apparent difference between the
methods of propionate formation in the rumen and the human large intestine
is not clear. It is of some interest to note that propionate formation is absolutely
essential to the ruminant because propionate is the only gluconeogenic volatile
acid in the rumen; i.e., it is the only one the animal can use for the synthesis of
The Rumen Fermentation 67

Table VI. Requirement for Vitamin B12 for Propionate Formation

by Human Intestinal Bacteroidesa
b
+ Vitamin B12 - Vitamin B 12

Source and organism Succinate Propionate Succinate Propionate

mol/IOO mol hexose c

Human intestine
Bacteroides eggerthii 79 27 115 0
B. /ragilis 33 49 92 9
B. splanchnicus 33 79 80 0
Rumen
B. succinogenes 77 0 n.d. d n.d.
B. ruminicola 130 0 n.d. n.d.
Ruminococcus flavefaciens 68 0 n.d. n.d.

a Miller and Wolin (1979); reprinted by permission.

b20 ng/ml.
cGlucose except for R. j7ave/aciens, which received ceUobiose.
dn.d., Not determined.

glucose. Glucose is as important in a ruminant's metabolism as it is in the

metabolism of monogastric animals. There is no evidence that propionate pro-
duction in the large intestine is of any direct significance to humans. Perhaps
the two-organism system is more effective as a propionate-producing system and
has evolved because of the essential role of propionate in ruminant metabolism.

6.4. Methanogenesis

Various aspects of methanogenesis have been reviewed recently, including

microbiological, biochemical, and ecological perspectives (Mah et al., 1977;
Zeikus, 1977). A few comments will be added here to emphasize some differ-
ences between the rumen and other ecosystems and to emphasize the role of
methanogenesis in maintaining low partial pressures of hydrogen in the rumen
and other systems.
Two major pathways are used in nature for the formation of methane. One
involves the reduction of carbon dioxide by hydrogen, and the other involves
the decarboxylation of acetate to methane and bicarbonate. Methane is pro-
duced from the methyl group of acetate by the latter pathway, and carbon
dioxide is not reduced. Not only is the carbon of methane derived from the
methyl group of acetate, but the three hydrogens of the methyl group are trans-
ferred intact to the final product. The contributions of carbon dioxide reduction
by hydrogen and acetate decarboxylation to methanogenesis in an ecosystem
68 Meyer J. Wolin

can be determined by measuring the amount of methane formed from radio-

active carbon dioxide and the amount formed from radioactive, methyl-labeled
acetate. Labeling studies in the rumen show that all the methane is produced
by reduction of carbon dioxide. Decarboxylation of acetate does not occur. In
waste decomposition and anaerobic sediments, about 70% of the methane
produced is derived from the methyl group of acetate and 30% is from reduction
of carbon dioxide (Mah et aZ., 1977). The probable reason for the lack of pro-
duction from acetate in the rumen is the slow growth of the bacteria that pro-
duce methane from acetate. Their generation times appear to be too long to
permit their establishment in light of the dilution rate of the rumen system. In
the other environments, however, turnover is much slower, and the acetate-
using organisms are able to grow and produce methane.
Production and loss of methane is usually considered as a loss of energy for
the ruminant. Approximately 10 to 15% of the energy of ingested food is lost
by eructation of methane. It is possible to imagine a fermentation that would
permit the accumulation of acetate, propionate, and carbon dioxide (like the
glucose fermentation of the propionibacteria) that would be more beneficial
to the animal. No energy would be lost as eructated gas, and the volatile acids
are those the animal already uses. Any interference with the production of
methane that leads to the accumulation of hydrogen, however, would not be
beneficial because there would be an even greater loss of energy. The reduction
of carbon dioxide by hydrogen does provide energy for the synthesis of the
protein and other cellular constituents of the methanogens which are of benefit
to the animal. Nevertheless, considerable attention has been given by animal
nutritionists to the possibility of using inhibitors of methanogenesis with the
hope of capturing the lost gaseous energy in nongaseous fermentation products
the animal can use advantageously (Demeyer and Van Nevel, 1975).
The use of hydrogen by methanogens is extremely important in all methano-
genic fermentations. The capacity of methanogens to use hydrogen is immense.
Methane is produced by rumen contents at a rate of approximately 5.5llmol/g
per hour (Carroll and Hungate, 1955). Since 4 mol of hydrogen are used to form
1 mol of methane, the rate of hydrogen utilization is 4 times the rate of produc-
tion of methane. The hydrogen-production rate is the same as the hydrogen-
utilization rate. (The amount of methane produced and hydrogen used per day
in a 500-kg cow is approximately 200 and 800 liters, respectively.) No hydrogen
accumulation occurs in the rumen; the partial pressure of hydrogen is approxi-
mately 10-4 atm (Hungate, 1967). (The partial pressure of methane is about
0.35 atm.) Very low partial pressures of hydrogen are also found in other
environments where methane is a major product of fermentation. The partial
pressure of hydrogen has an important influence on the course of the rumen
fermentation and other methanogenic fermentations.
The Rumen Fermentation 69

6.5. Hydrogen Gas and Fermentation

Some bacteria are able to form hydrogen from reduced pyridine nucleotides
(pNH), but the accumulation of hydrogen inhibits the reaction. The equilibrium
constant strongly favors the reverse reaction, the production of PNH from oxi-
dized pyridine nucleotides and hydrogen. Removal of hydrogen from the en-
vironment, however, permits the expression of hydrogen formation from PNH.
Other major mechanisms for microbial formation of hydrogen include formation
from formate and pyruvate. These routes are not susceptible to reversal by
hydrogen. The importance of the hydrogen-inhibited and -uninhibited routes on
the fermentation of glucose can be illustrated by the fermentation pathway of
R. aZbus (Glass et aZ., 1977) (Fig. 5). It ferments glucose by the Embden-
Meyerhof-Pamas pathway to pyruvate and NADH. Acetyl-CoA, hydrogen, and
carbon dioxide are formed from pyruvate in a ferredoxin-dependent reaction.
The catalytic amounts of NADH produced in the cell must be reoxidized. They
can be reoxidized by reduction of acetyl-CoA to acetaldehyde, which, in tum,
is reduced to ethanol by NADH. The acetyl-CoA that is not reduced is converted
to acetate. These reactions lead to a balanced fermentation of glucose to ethanol,
acetate, hydrogen, and carbon dioxide. R. aZbus can also reoxidize NADH with
the production of hydrogen with another ferredoxin-dependent system but only
when hydrogen is removed from its surroundings. This happens in the rumen
when the organism is growing in the presence of methanogens or in artificial
systems when the organism is cocultured with a hydrogen-using species (Iannotti

---NAD
=l Ethanol + GoA

ADP. Pi ATP, GoA

>-_~:~,~.~oA
A""~c:_........... A=-t~~AGETATE ..;;;::::o_ _

Glucose
Fd
+
NAD~Fdr:Xd H

- - . NADH Fd H2

Figure s. Pathway of fennentation of glucose by Ruminococcus albus. Fd, Ferredoxin.

From Glass et al. (1977); reprinted by permission.
70 Meyer J. Wolin

et al., 1973). If NADH is used to produce hydrogen, it cannot be used to pro-

duce ethanol. All of the acetyl-CoA fonned from pyruvate becomes available
for the production of acetate. When hydrogen is removed, the balanced fermen-
tation products are acetate, carbon dioxide, and the removed hydrogen, and
ethanol is no longer a product. Ethanol is not produced from carbohydrate in
the rumen (Moomaw and Hungate, 1963), despite the fact that important species
such as R. albus produce ethanol as a major fermentation product in pure
cultures (where hydrogen is usually allowed to accumulate).
Similar effects of hydrogen utilization on the fonnation of succinate by R.
flavefaciens (Latham and Wolin, 1977) and propionate and lactate by S. rumi-
nantium (Chen and Wolin, 1977) have been demonstrated. The general principle
is the same as for the R. albus fennentation. Succinate, propionate, and lactate
are electron-sink products of the respective carbohydrate fennentations of
R. flavefaciens and S. ruminantium, as is ethanol for R. albus. They can reoxi-
dize NADH fonned during glycolysis by fonning the electron-sink products,
succinate, lactate, propionate, or ethanol, from pyruvate. When the partial
pressure of hydrogen is kept very low by its removal by a methanogen, produc-
tion of any of the electron-sink products greatly diminishes, and the production
of acetate from pyruvate increases.
More detailed discussions of these interactions are available in other publica-
tions (Wolin, 1974, 1975, 1976), and only a few highlights will be discussed
here. The organisms mentioned above have alternative mechanisms for reoxidiz-
ing NADH, and their fermentation products will differ depending upon the par-
tial pressure of hydrogen in the environment. Some organisms are known that do
not use carbohydrates and do not have alternatives. They can only reoxidize
NADH by fonning hydrogen and can only do so when the partial pressure of
hydrogen is kept low. These organisms include S organism (Reddy et al.,
1972a) and desulfovibrios (Bryant et al., 1977). They can obtain energy for
growth by oxidizing ethanol to acetate and hydrogen with PNH-dependent
enzyme systems, and desulfovibrios can similarly use lactate. Significant growth
occurs only when the organisms are cocultured with a hydrogen-using organism
(a methanogen). Alternatively, desulfovibrios can grow by themselves if they are
provided with sulfate in addition to ethanol and lactate. Organisms that depend
on interactions with methanogens for growth do not appear to be important in
the rumen ecosystem. They appear to be more important in the complete bio-
conversion systems for producing methane found in anaerobic sediments,
swamps, and in anaerobic waste decomposition. It is possible that the dynamic
balance between hydrogen fonnation and utilization in the rumen normally
avoids the fonnation of pulses of products such as ethanol and lactate, which are
energy sources for the growth of organisms like S organism and desulfovibrios.
Closer juxtaposition between interacting organisms, better mixing, and the
The Rumen Fermentation 71

generally greater constancy of nutrients and the environment of the rumen may
mitigate against the more complex food chains of the other ecosystems.
In addition to the interactions between S organism or desulfovibrios and
methanogens, interactions such as those described for the rumen organisms
probably occur in concert with the dependent interactions in other ecosystems.
Carbohydrate-fermenting clostridia can form hydrogen from NADH (Junger-
mann et al., 1973), and interactions between a thermophilic, cellulolytic Oostri-
dium and a thermophilic methanogen (Weimer and Zeikus, 1977) are similar
to those described for R. albus. Interaction between hydrogen formation and
utilization is not necessarily restricted to hydrogen utilization by methanogens.
The reduction of carbon dioxide to acetate is an important mechanism for
using hydrogen in the ceca of rodents (prins and Lankhorst, 1977), and sulfate
reduction by hydrogen by sulfate-reducing bacteria (Badziong et al., 1978) is
another potentially important mechanism for the depletion of hydrogen in
anaerobic ecosystems. Production of hydrogen from NADH is also not restricted
to carbohydrate-, ethanol-, and lactate-using microorganisms. The products of
Gly fermentation by Diplococcus glycinophilus are altered by the partial pres-
sure of hydrogen in a manner that is consistent with an effect on alternative
modes of oxidizing NADH (Cardon and Barker, 1947; Wolin, 1976). The use of
formate and probably pyruvate by S organism involves reoxidation of NADH to
hydrogen (Reddy et al., 1972b; Wolin, 1976).
Additional research is necessary to obtain a clearer perspective of the role of
hydrogen formation and utilization on the metabolism of individual micro-
organisms and on the flow of carbon in anaerobic ecosystems. It is clear, how-
ever, that these interactions can profoundly influence the types of products
formed in the environment and, therefore, the actual substrates available for the
establishment of specific food chains and the relevant microorganisms. For
organisms that do not possess alternative routes of oxidation of NADH, inter-
action is essential for the production of energy and for their survival. Those
organisms that do have alternatives may obtain more energy for themselves
when they produce acetate rather than electron-sink fermentation products
from pyruvate. For example, more ATP is produced from glucose when acetate
rather than ethanol or lactate is the fermentation product. The increased ATP
yield may be advantageous to an organism if the amount of its energy supply
in its environment limits its growth.

6.6. Growth Rate and Fermentation

It is difficult to analyze the kinetics of growth of individual populations in

the rumen ecosystem. Cognizance of the general subject of growth kinetics
indicates, however, that there are many factors in a complex ecosystem that can
72 Meyer 1. Wolin

control the rate of growth of microogranisms. These factors include the rates of
hydrolysis of polymeric carbon and nitrogen sources, concentrations of the
permeable macronutrients (e.g., carbon, energy, and nitrogen sources) actually
used by the organisms, concentrations of any potentially growth-limiting micro-
nutrients (e.g., vitamins and trace metals), inhibitory agents, and pH. Since
protozoa depend to a significant extent on ingestion of intact bacteria for nutri-
ents, rates of ingestion and intracellular digestion are important in determining
the rates of growth of protozoa. Various aspects of the effects of growth rate,
substrate concentration, and pH on fermentation have been reviewed (Hobson
et al., 1974). One example of the influence of growth rate on fermentation is
the demonstration that S. rnminantium produces mainly propionate, acetate,
and carbon dioxide from glucose when grown at slow growth rates in a glucose-
limited chemostat but switches to the production of increasing amounts of
lactate as the growth rate increases (Hobson and Summers, 1967; Scheifmger
et al., 1975). These pure culture experiments suggest that the fermentation of
S. rnminantium in the ecosystem may vary depending on the rate it is supplied
with a source of fermentable carbohydrate. This example is used to illustrate
the point that many factors other than the partial pressure of hydrogen can alter
the course of fermentation of individual species. Since it is difficult to directly
measure growth rate and the environmental parameters that influence it in the
ecosystem, studies of factors that regulate pure cultures can at least help define
their potential for carrying out different kinds of fermentations and the condi-
tions that might regulate the fermentations in the ecosystem.

6.7. The Overall Fermentation

The fermentations of individual populations and the already described inter-

actions between populations produce acetate, propionate, butyrate, methane,
and carbon dioxide from plant carbohydrates. The stoichiometry of the fermen-
tation of glucose, the monosaccharide unit of cellulose and starch, by the
community has been deduced by independent methods (Wolin, 1960; Hungate,
1966) and is approximately

57.5 (C 6 H12 0 6 ) ---+ 65 acetate + 20 propionate + 15 butyrate + 60 CO2

+ 35 CH4 + 25 H20
The major pathway for hexose fermentation in the rumen is the Embden-
Meyerhof-Parnas pathway, the major pathway of most of the individual species
of carbohydrate-fermenting rumen microorganisms. This subject and the path-
ways of pyruvate metabolism and product formation of carbohydrate-fermenting
rumen bacterial species have been reviewed recently (Miller and Wolin, 1979).
As previously discussed, hydrogen utilization would tend to decrease the amount
The Rumen Fermentation 73

of propionate and eliminate the lactate and ethanol formed by isolated single
species with corresponding increases in acetate in the ecosystem. There is no
evidence that hydrogen utilization influences the formation of butyrate in the
rumen or by pure cultures of one of the major butyrate-forming species, B.
fibrisolvens (Wolin, unpublished data). It should be emphasized that the con-
centrations of individual species in the rumen may also be a major factor in
determining the proportions of products formed from carbohydrate. For exam-
ple, B. succinogenes produces succinate and acetate from cellulose but does
not produce hydrogeti:. ·Conditions that favor B. succinogenes over the celluloly-
tic R. albus would favor the production of propionate (by decarboxylation of
succinate) rather than production of methane and acetate (by R. albus; see Fig.
5). The relationship between fluctuations in the concentrations of individual
species and the relative proportions of acid products has not been explored.

7. Summary and Conclusions

This review has focused on the rumen fermentation. In order to discuss the
fermentation, it was necessary to provide an overview of the ruminant as a
fermentation factory. This was possible because of the vast amount of available
information about ruminant physiology, metabolism, and nutrition. Analogous
information about other intestinal-tract systems is scarce, and relationships
between microbial activities in the intestinal tract and host physiology, metabo-
lism, and nutrition are not always clear. It was also possible to discuss the in vivo
rumen microbial community and its activities, gross composition, environment,
and dynamics because of the studies of many investigators. Again, there is a
scarcity of similar studies of other intestinal-tract and other anaerobic eco-
systems. Finally, it was possible to attempt to dissect the activities of the rumen
community into activities of individual populations of bacteria and protozoa
because many of them have been isolated and well characterized. Again, this is
not always true for other intestinal-tract or other complex anaerobic ecosystems.
The rumen fermentation is of special importance to man. Domestic rumi-
nants are factories for the conversion of solar energy, through the medium of
plants, into very high-quality food, fiber, and leather. Ruminants can thrive on
plants that are otherwise difficult to harvest and to process into something
useful for man. Microorganisms in the rumen convert the cellulose and other
carbohydrates, the nitrogen constituents, and trace elements of plants to short-
chain volatile fatty acids and microbial cells. The animal uses the microbial
products as sources of carbon, energy, protein, and vitamins. The ruminant and
its microbial community interact to provide the proper environment for the
necessary conversions. The microbial populations sustain each other by provid-
ing each other with essential nutrients.
74 Meyer J. Wolin

The rumen fennentation itself is the result of interactions between many

different microbial species. These interactions are not always apparent from
conventional pure-culture studies-e.g., the ability of noncellulolytic organisms
to use soluble oligosaccharides formed from cellulose by cellulolytic species,
the decarboxylation of succinate produced by one species by another species,
and the changes in fennentation products caused by the use of hydrogen by
methanogenic bacteria.
The rumen system warrants further study not only because of its practical
importance but also because it is an excellent system for studying phenomena
that are of importance in other microbial ecosystems. Because of the existing
data base and the remarkable constancy of the environment, the rumen
microbial community is more amenable than most to studies of microbial
interactions in complex ecosystems. We should, however, increase our efforts
to analyze other systems using the strategies that have been successful for studies
of the rumen. As with all complex ecosystems, it is clear that it is necessary to
understand how the whole system operates in order to understand the contribu-
tion of its parts and that it is necessary to understand the contribution of the
parts in order to understand the whole. Other systems may not be as well
controlled as the rumen system and, therefore, less easily studied. It would
probably be useful, however, to recognize that integration of studies of the
community as a whole with studies of the populations of the community is not
only a necessary but also a synergistic way of obtaining an understanding of the
activities of an ecosystem.

ACKNOWLEDGMENT

The author's research is supported by research grant number AI-12461,

awarded by the National Institute of Allergy and Infectious Diseases, U.S. Public
Health Service, Department of Health, Education, and Welfare.

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