PRODUCTIVE LIMITS TO RUMEN F E R M E N T A T I O N 1,2

Werner G. Bergen 3 and Melvin T. Yokoyama 4

Michigan State University, East Lansing 48824

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SUMMARY polysaccharides into usable substrates and the
concomitant fixation of non-protein nitrogen
This paper is a review of the current knowl- into microbial cells places the ruminant in a
edge on the productive limits to rumen fermen- unique niche in animal production systems. The
tation. The following topics are discussed: trend has been toward ever-increasing output of
Ruminal carbohydrate fermentation and ATP milk and meat by ruminants. The question that
generation; heat and gas losses during ruminal must be considered is to what extent can the
fermentation; ruminal microbial growth yields rumen provide nutrients to the host for high
and efficiency; YATP and the maintenance performance or should the rumen be regulated
coefficient; manipulation of fermentation prod- or bypassed to get more efficient utilization of
ucts and growth efficiency; digesta passage and more refined feedstuffs. The anerobic nature
substrate disappearance; the role of ciliate
and other characteristics of the ruminal fermen-
protozoa in the rumen. This review reempha-
tation place an upper limit on the potential
sizes the need for even further study to enhance
nutrient yield for the host from this fermenta-
the utilization of feedstuffs for optimal animal
tion. In this paper, we will describe the physio-
production.
logical processes that limit ruminal output of
(Key Words.- Rumen, Fermentation, ATP Yield, nutrients and discuss how these processes may
YATP, Cell Yield, Maintenance Coefficient.) be regulated to maximize ruminal output.
INTRODUCTION
Ruminal Carbohydrate Fermentation and
The feedstuffs consumed by ruminants are ATP Generation. The occurrence of an aerobic
all initially exposed to the fermentative activity fermentation in the digestive tract of the
in the rumen. Dietary polysaccharides and ruminant dictates that besides significant inher-
protein are generally degraded by the ruminal ent energy losses (i.e., heat of fermentation and
microorganisms into characteristic end products methane), the amount of free energy (ATP)
which in turn provide nutrients for the host's derived from the dietary constituents consumed
metabolism. The extent and type of transfor- will only represent a fraction of its aerobic
mations of feedstuffs in the rumen will influ- potential. Stoichiometric estimates indicate
ence the host's productive performance. Hun- that only 4 to 5 ATP are generated per hexose
gate (1966) has described the rumen as a molecule fermented in the rumen representing
common denominator and also as a limiting about 10 to 12% of its potential aerobic yield
process in agricultural production systems with (Hungate, 1966). This generated ATP, however,
ruminant animals. is not entirely lost to the animal, since it is put
Clearly, the ruminal degradation of 3-1inked to use for de novo synthesis and maintenance
of microbial cells, which along with the volatile
fatty acid end products of fermentation can be
1Michigan Agricultural Experiment Station Journal utilized by the animal to meet its nutritional
Article No. 7906. requirements. The resultant effect of this ar-
2Presented at the Symposium on Turning Rumen
Fermentation On and Off to Maximize Animal Pro- rangement, however, is that productive effi-
ductivity, Northeast Sections of the American Society ciency of the animal is compromised and
of Animal Science and American Dairy Science Associ- probably limited, in exchange for sustaining a
ation, July 12, 1976, Beltsville Agriculture Research rumen microbial capacity to convert fibrous
Center, MD.
feeds and non-protein nitrogen into products of
3Ruminant Nutrition Laboratory, Department of
Animal Husbandry. nutritional value. The key question, therefore,
4Agricultural Fermentation and Nutrition Labora- appears to be how to manipulate ruminal
tory, Department of Animal Husbandry. fermentation such that the optimal production
573
JOURNAL OF ANIMAL SCIENCE, Vol. 46, No. 3, 1977
574 BERGEN AND YOKOYAMA
of end products of nutritional value is coupled
with an optimal response in net microbial cell
growth.
The chief source of energy (ATP) in the
rumen is derived from the fermentation of
carbohydrates (starch, cellulose, pectins and
hemicellulose) to volatile fatty acids. While the
basic ruminal microbial saccharoclastic path-
ways have been elucidated (Baldwin, 1965),
estimates of the theoretical yield of ATP from
specific reactions are still arbitrary, especially
for propionate and methane production. In the
rumen, propionate is produced via the dicar-
boxylic acid (randomizing) pathway and by the
direct reductive (non-randomizing) pathway.
The contribution of each pathway is dependent
on the diet. While the dicarb9xylic acid path-
way is the predominant one, increasing carbo-
hydrate availability will increase the role of the
direct reductive pathway, presumably because
of an increase in lactate as a precursor (Baldwin
et al., 1963). While the theoretical ATP yield
(.67 mol ATP/mole lactate) for the conversion
of lactate to propionate appears to be in
agreement for various propionate-producing
microorganisms, the theoretical ATP yield for
the conversion of glucose to propionate is still
in doubt. It has been suggested (Hobson and
Summers, 1972) that Selenomonas rumina-
ntium in the conversion of pyruvate to propio-
nate, with glucose as substrate, derives 3 ATP in
the flavoprotein-linked electron transport from
NADH to fumarate, pyruvate to malate, and
methylmalonyl CoA to propionate. This is not
consistent in propionibacterium, which forms
propionate from pyruvate via malate dehydro-
genase and methylmalonyl CoA transcarboxyl-
ase. For propionibacterium it is postulated that
2 ATP are generated in the cytochromeqinked
electron transport from NADH to fumarate
(de Vries et al., 1973). Selenomonas ruminant-
ium, however, could account for most, if
not all, of the rumen production of propio-
nate via succinate. (Wolin, 1975), and its
decarboxylating activity is 87 times as great as
that of propionibacteria (Sijpesteijn and Elsden,
1952).
The energetics of methane production are
still not clearly understood (Stadtman, 1967).
Calculations of the free energy change for the
reduction of CO2 with molecular hydrogen to
methane is about - 3 2 Kcal per mole of
methane formed. Based on this value, a theoret-
ical yield of 4 ATP per mole CH4 produced can
be calculated. However, based on a YATP value
of 10, as determined for Metbanosarcina bakeri
and "Metbanobacterium omelianskii" (S-
organism and MOH strain) values between .16
and .44 mole of ATP per CH4 are obtained
(Stadtman, 1967). ATP is required for initial
activation of the reaction, with the terminal
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methyl reduction probably being the site of
free energy exchange. A current assignment of
moles of ATP generated per mole of end
product produced is: acetate, 2; propionate, 3;
butyrate, 3; and methane, 1; (Is~acson et al.,
1975).
The most important factor which influences
ruminal microbial fermentation is the diet.
Alteration of the rate and type of fermentation
which ensues by manipulation of dietary carbo-
hydrates and their physical form is a well
known phenomenon. However, the nature of
the change in terms of the microbial population
and its energetic consequences is not clearly
understood. Furthermore, our knowledge of
the modifying effects of factors such as dilution
rate (rate of passage) and intermicrobial interac-
tions, especially on YATP yields, is incomplete.
While shifts in ecological niches constantly
occur in the rumen, resulting in transit changes
in the microbial population (adaptation), some
pure culture studies (Wolin, 1975) suggest that
fermentation end products could be quite
different for specific ruminal microorganisms
depending on the effect of the carbohydrate
substrate on dilution rate. Other studies (Hob-
son, 1965; Stouthamer and Bettenhaussen,
1973) have demonstrated that microorganisms
will shift their fermentation to less efficient
pathways as dilution rate increases, but because
of lower maintenance requirements, show a net
growth response. There are instances however,
of microorganisms which do not adhere to this
mechanism of action (Wolin, 1975). Some
recent continuous culture studies show that
different dilution rates do not affect total VFA
or acetate production, but as the dilution rate
increases, propionate tends to increase and
butyrate tends to decrease (Isaacson et al.,
1975). In this regard, an in vivo increase of
dilution rate by incorporation of mixed mineral
salts into a diet resulted in the increased
dilution rate being negatively related to the
molar proportion of propionate and positively
related to the molar proportion of acetate, with
no difference in total VFA production (Thomp-
son et aL, 1975).
While the rumen is the major site of fermen-
tation, and our understanding of its function
 RUMEN FERMENTATIONTO MAXIMIZEANIMALPRODUCTIVITY
(albeit incomplete) permits us to discuss its
manipulation, much remains to be learned
about post-ruminal fermentation and the micro-
biology of the ruminant large intestine. A
recent review of the structure and function of
the ruminant large intestine has been published
(Ulyatt et al., 1975). Providing large amounts
of fermentable carbohydrates has demonstrated
that while most of it will be fermented in the
rumen, a significant quantity will escape to the
lower gut and undergo secondary fermentation
(Orskov et al., 1971; Karr et al., 1966).
Considerable amylolytic, cellulolytic, proteo-
lytic, deaminase and urease activity has been
detected in the large intestine (Orskov et al.,
1969, 1970; Hecker, 1971, Mann and Orskov,
1973). Commensurate with this fermentation
are changes in the microbial population, and
the production of additional volatile fatty acids
and microbial cells. Overloading concentrates
into the rumen of cattly and sheep has been
shown to increase caecal concentrations of
lactobacilli, streptococci, coliforms and Clos-
tridium perfringens (Allison et aL, 1975). While
caecal and large intestine fermentation and
microbial cell synthesis would appear to be a
disadvantage to the ruminant, there is evidence
for volatile fatty acid and ammonia absorption
Ulyatt et al., 1975), the significance of which
has not been fully ascertained. In view of recent
interest in rumen by-pass of nutrients and
increasing dilution rates, however, it would be
an important consideration to be taken into
account. The nutritional implications of the
autochthonous microflora of the lower intesti-
nal tract of the ruminant has yet to be studied.
Heat and Gas Losses During Ruminal Fer-
mentation. Actual energy losses as a result of
ruminal fermentation far exceed the energy
losses associated with the hydrolytic reactions
of nonruminant digestion. While short of at-
tempting to completely remove this ruminal
fermentation in the process of digestion, the
question arises as to how, and to what extent
these energy losses can be reduced to improve
productive efficiency of the animal, and yet
retain the advantage of the arrangement. The
ruminal energy losses which bear the most
scrutiny in this regard are (1) the heat of
fermentation, and (2) methane production.
The heat of fermentation is the free energy
which is dissipated as a result of inefficiencies
in microbial metabolic activity (anabolic and
catabolic reactions) in the rumen. By defini-
tion, it is not associated with the energy loss in
575
maintenance of the microbial population. Only
a few direct calorimetric estimates have been
made of this energy loss. Heat of fermentation
(HF) values for two cows immediately after
consuming a concentrate feed were .99 and .77
Kcal/kg ingesta/hr. After being fed a mixed
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grass-legume hay ration, the HF values were .44
and .38 Kcal. Fasting for 24 hr, resulted in
decreased HF values of .10 and .11 Kcal
(Houp~, 1968). Indirect estimates (CO2/CHa
ratio, in vitro substrate fermentation and stoi-
chiometric calculations of end products) indi-
cate that the heat of fermentation ranges from
3% to 12% of the gross energy of the feed
(Blaxter, 1962), with a probable modes for
cellulose of about 6% (Marston, 1948). This is
in contrast to a value of about 1% for digestive
heat loss in nonruminants. It is still open to
question as to how effectively this energy loss
can be recovered, and how significant a contri-
bution it would make to productive efficiency.
Factors such as feed quality and quantity will
have a marked influence on its variability, by
affecting the type of fermentation developed
(Blaxter, 1962). Accommodations can also be
made in reducing fermentative heat loss by
altering the physical form of dietary constitu-
ents (grinding, flaking, chemical predigestion,
etc.), but overall conservation must be assessed
against the rate of fermentation and microbial
growth response. Furthermore, fermentative
heat can be and usually is credited to body heat
maintenance (Houpt, 1968). To the extent that
it would reduce fermentative heat loss, selective
protection of dietary components (partial
avoidance of fermentation) could also be a
manipulative approach. This would however,
circumvent rather than alleviate the problem.
Thus, the problem appears to be that since the
heat of fermentation is an exponential expres-
sion of total microbial energetic efficiency
(Walker, 1965), it is difficult to take into
account the numerous variables and their inter-
actions, some of which have yet to be even
identified in the rumen. In this regard, the
utilization of rumen simulation modeling sys-
tems, as proposed by Baldwin et al. (1970), and
by Koong and Baldwin (1976) would be of
benefit in rapidly evaluating microbial interac-
tions and the energetic efficiency of fermenta-
tion types.
The production of methane in the rumen is
subject to fewer variables than fermentative
heat, and as such would appear to be more
amenable to direct manipulative control. The
576 BERGEN AND YOKOYAMA
identification of these variables has been largely
due to the intensive research interest which has
proceeded in this area, and several excellent
reviews (Bryant, 1965; Stadtman, 1967; De-
meyer and Van Nevel, 1975) are available.
Methane will usually constitute between 15% to
30% of total ruminal gases, and its production
is influenced by such factors as the physical and
chemical nature of the feed, as well as the level
of intake (Hungate, 1966). In contrast to
fermentative heat, however, its production
(e.g., energy loss) expressed as a percentage of
the gross energy of a particular feed is relatively
constant. A reasonably accurate estimate of its
energy loss from most feeds is about 8% of the
gross energy (Blaxter, 1962). Another distinct
characteristic of ruminal methane production is
that it is not directly proportional to the
digestibility of the feed consumed (Blaxter,
1962). Highly digestible feeds result in less
methane production per unit of caloric energy
consumed. However, above a maintenance level
of intake, this decrease is less for these feeds
than for feeds of a lower digestibility (Blaxter
and Clapperton, 1965). Ratios for moles of
methane produced to moles of hexose fer-
mented for various carbohydrate substrates
range from .3 to .5 with a mean of .45
calculated from a number of in vivo determina-
tions (Hungate, 1966).
It is now well established that the major
precursors of rumen methane are H 2 and CO2.
Formate, which is degraded to CO2 and H2 via
formate hydrogenlyase, is estimated to contrib-
ute about 18% of rumen methane (Hungate et
al., 1970). Other precursors include methanol
and acetate, other alcohols, and volatile fatty
acids. While these other precursors are not
thought to be significant in the rumen, acetate
may be important under certain conditions.
Earlier studies showed that rumen enrichment
cultures produced methane from acetate and
suggested that its contribution to methanogene-
sis may increase in vivo with time after feeding
(Opperman et al., 1961). Data from in vitro
continuous culture studies with ruminal micro-
organisms suggest that acetate may be a signifi-
cant precursor of methane, especially at low
dilution rates (Isaacson et al., 1975). Under
conditions of low partial pressures of hydrogen
and high acetate concentrations, acetate may be
the preferred precursor for methane produc-
tion, since less reducing equivalents would be
required for reduction of the methyl group
than for carbon dioxide (Zeikus et al., 1975).
Actually very little is known about the role o f
acetate in rumen methanogenesis, and rumen
methanogens which utilize it as a precursor.
Nutritional studies with Methanobacterium
ruminantium, have identified an essential
growth factor, 2-mercaptoethanesulfonic acid
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(coenzyme M), for the organism (Taylor et al.,
1974). Other work (Prins, 1974) has demon-
strated an N-acetylglucosamine requirement for
Mb. ruminantium. These findings suggest an
opportunity for controlling rumen methano-
genesis by this specie.
Ruminal Microbial Protein Synthesis. The
Role o f Dilution Rate and Maintenance Coeffi-
cient on the Efficiency o f Cellular Growth. The
ruminal fermentation can be likened to a
continuous, anaerobic microbial culture
(chemostat) system (Hungate, 1966). The
salient components of a continuous system,
substrate supply, end product removal, buffer
capacity, temperature control and dilution rate
(growth rate) regulation are all present in the
unique symbiotic relationship between the
reticulo-rumen and the host.
Two classes of microorganisms are usually
found under natural ecological conditions. The
aerobic organisms use excess oxygen as an
electron aceptor and thereby have a high
potential of ATP generation from a given
substrate. The anaerobic organisms must use a
variety of limiting metabolites as electron ac-
ceptors and hence have a much lower potential
for ATP generation from a given substrate
(Gunsalus and Shuster, 1961). The limiting
factors for microbial growth under aerobic and
anaerobic conditions are carbon and energy,
respectively (Gunsalus and Shuster, 1961).
Hungate (1966) proposed that energy availabil-
ity for microbial growth was the limiting factor
and hence there must be an upper limit of
microbial cell production in the rumen. Since
anerobic rumen microorganisms are a major
source of protein for the host and provide the
only mechanism for non-protein nitrogen
(NPN) utilization, and since high productivity
by the host is desirable, quantitative aspects of
microbial cell synthesis are important to animal
production.
Extensive research was conducted in batch,
energy limiting cultures to relate.cell growth to
substrate disappearance. Bauchop and Elsden
(1960) concluded that adenosine triphosphate
(ATP) was the common denominator between
growth and substrate disappearance and that
microbial growth was directly proportional to
 RUMEN FERMENTATION TO MAXIMIZE ANIMAL PRODUCTIVITY
the amount of ATP generated from the catabo-
lism of energy substrates. Bauchop and Elsden
(1960) defined the relationship between ATP
and cell growth as YATP (molar growth yield
per mole ATP or g organisms (dry weight) per
mole ATP). From the work of Bauchop and
Elsden (1960) and others (Forrest and Walker,
1971, Payne, 1970; Stouthamer, 1969), it was
further concluded that the YATP was a con-
stant value for different microorganisms. The
generally accepted value for YATP is 10.5
(Stouthamer and Bettenhaussen, 1973).
From the formulation of the YATP concept
and from knowledge of ATP yields for the
various catabolic pathways in the rumen, Hun-
gate (1966) calculated that the rumen fermen-
tation can produce about 10 g of microbial
protein for each 100 g carbohydrate fermented.
This value would represent an upper limit of
synthetic capacity in the anaerobic ruminal
fermentation. Purser (1970) calculated that this
level of protein synthesis was equivalent to 18.3
g digestible protein per Mcal digestible energy, a
protein-calorie ratio below that required to
meet the protein requirements of young grow-
ing ruminants and less than such ratios estab-
lished from empirical balance studies with
growing ruminants (Purser, 1970). In vitro
isotope incorporation studies with rumen con-
tents (Bucholtz and Bergen, 1973), energy
balance calculations from ruminal fermenta-
tions (Baldwin, 1970), and ingesta passage
studies with sheep using NPN-eontaining diets
or microbial cell markers (Hogan and Weston,
1971; Hume et al., 1970; Lindsay and Hogan,
1972) all showed that the microbial protein
synthesis rate was from 15 to 22 g per 100 g
organic matter fermented.
Growth yield studies with pure cultures of
rumen bacteria (Hobson, 1965, Hobson and
Summers, 1967; Hungate, 1963), certain Strep-
tococcus strains (Moustaffa and Collins, 1968)
and A c t i n o m y c e s israelii (Buchanan and Pine,
1967) produced YATP values from 15 t o 2 0 .
These high values were at that time considered
anomalous growth yields since the pathways for
ATP production were not adequately known
for the above organisms (Forrest and Walker,
1971).
Assuming a constant YATP (Payne, 1970),
the anomalous growth yields of pure cultures of
organisms and microbial protein yields in excess
of the upper limit proposed by Hungate (1966)
may be attributed to higher ATP production
than previously proposed. Extra ATP could be
577
generated from cytochrome-linked electron
transfer or substrate phosphorylation reactions
in anaerobes. (Hobson and Summers, 1 9 7 2 ;
Hungate et al., 1971 ; deVries et al., 1974).
Recently a major breakthrough was achieved
in this area when Stouthamer and Bettenhaus-
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sen (1973) proposed that YATpwas not con-
stant and that the molar growth efficiency of
microorganisms depends on their specific
growth rate and maintenance requirements.
These workers pointed out that all previous
studies on YATP were done in batch cultures or
continuous cultures with excess energy. Stou-
thamer and Bettenhaussen (1973) determined
the YATP for A. aerogenes grown at various
growth rates (dilution rates; D) in an energy
limiting chemostat. They were able to show
that as growth rate increased, YATP also
increased. The yA Max
TP was determined to be
around 25. From these results, Stouthamer and
Bettenhaussen (1973) were also able to esti-
mate the maintenance coefficients at various
growth rates. It was shown that slow growing
organisms have a much higher maintenance
coefficient and it was concluded that any given
YATP value is determined by the specific
growth rate (or more generally dilution rate; D)
and maintenance coefficient (Me) in any given
physiological situation. Thus YATP is low
(<10) in slow growing cultures but may ap-
= Max
proach the YATP of 25 at high growth rates.
Major maintenance energy needs by microorga-
nisms would be for motility, replacement of
lysed cells, intracellular ionic concentrations,
active transport and turnover of intracellular
components (Marr et al., 1963). It is only after
these requirements have been met that net
microbial growth from which the animal can
benefit can occur.
Isaacson et al. (1975)extended the approach
of Stouthamer and Bettenhaussen (1973) to
mixed ruminal cultures grown at various D in a
continuous culture system using glucose as an
energy substrate. These workers found that the
YAT P was also not constant for ruminal bacte-
ria and observed that YATP rose from a value
of 7--8 at a D of .02 h-m to 1 6 - 1 7 at a D of .12
h q . lsaaeson et al. (1975) found that at a low
D, approximately 55% of the energy derived
from glucose was used for maintenance, where-
as at high D only 15% of the available energy
was used for maintenance. These workers con-
cluded that over the physiological range of
ruminal conditions, YATP values vary broadly
 578 BERGEN AND YOKOYAMA
and are dependent on the maintenance coeffi-
cient of the bacteria, which is a function of the
dilution rate.
Cole e t al. (1976) and Kropp e t al. (1976)
studied abomasal passage of microbial protein
and ruminal organic matter disappearance.
They used various combinations of roughage,
grain, and protein supplements in their studies.
Based on lignin passage to the abomasum, Cole
e t al. (1976) and Kropp e t al. (1976) also
computed ruminal dilution rates. In these stud-
ies, dilution rates varied from .02 to .06 h -1.
Microbial protein synthesis was higher at the
higher dilution rates and the overall range was
from 7 to 10 g per 100 g DOM to 17 to 20 g
per 100 g DOM. These results are a direct
example of the principles outlined by Stouth-
amer and Bettenhausen (1973) and Isacson e t
al. (1975) that efficiency of microbial growth is
dependent on maintenance which in turn is a
function of dilution rate. The extra VFA
produced by the microorganisms during periods
of high maintenance needs are not totally lost
as they can be used by the host; however this
use will have a negative effect on the protein-
energy ratio as more energy is available con-
comitantly with less microbial protein produc-
tion (i.e., less microbial cells mass produced per
unit of energy and per unit of substrate). As
previously indicated, Me is not a constant and
will vary with different microorganisms. The
Me coefficient is also not a fixed entity for a
specific microorganism, since conditions of
growth such as excess or limited substrate
availability, osmolarity, and redox state will
affect its value (Stouthamer and Bettenhaussen,
1973).
Other factors (e.g., other than D) may also
influence the maintenance coefficient of micro-
bial cells and hence modulate growth efficiency
and the YATP. Stouthamer and Bettenhaussen
(1973) reported that a shift in media NH4 CI
levels (from .2 to .4%) increased the mainte-
nance coefficient in A . a e r o g e n e s but the V~ATP
ax
was not affected. Other factors that may affect
microbial maintenance requirements are the
presence of growth inhibiting substances and
the unfavorable shifts in the ionic composition
in the culture media (Stouthamer and Betten-
haussen, 1973). Since there are wide fluctua-
tions in ruminal osmolality from hypotonic
(250 m osmolar) to hypertonic (400 m os-
molar) after feeding (Bergen, 1972), the effi-
ciency of microbial growth could be impaired.
This whole area needs further study in ruminal
systems.
Under ruminal conditions, the diurnal varia-
tions in microbial numbers and fermentation
activity in relation to time after feeding (War-
ner, 1965) suggest that fluctuations in the
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microbial Me coefficient could be quite large.
Increasing the frequency of feeding should
stabilize these fluctuations. Increasing feed in-
take, on the other hand, generally means a
higher rate of passage (increased dilution rate)
(Van Soest, 1975) which will increase YATP
and Y substrate yields by decreasing the micro-
bial maintenance. Serious attempts to manipu-
late rate of passage should take into account:
(1) a decreased feed digestibility, ( 2 ) a decrease
in ruminal protozoa numbers, (3) possible
changes in macromolecular composition of mi-
crobial cells, and (4) possible changes in fer-
mentation end products. Optimal microbial cell
yield must be assessed and counterbalanced
against the digestibility of the feed, as an
increase in cell yield (especially with highly
fermentable carbohydrates) would not be ex-
pected to nutritionally offset a decrease in
digestibility (Isaacson e t al., 1975). This factor
again reemphasizes our need to thoroughly
understand the function of the lower intestinal
tract of the ruminant in regard to nutrient
absorption and energetic efficiency. With an
increased rate of passage, ruminal protozoa
numbers will also decrease (Coleman, 1975)
and considerations must be made for the
lowered amount of protein from this source.
There is, however, some question as to the
significance of ruminal protozoa contribution
to net microbial protein passage, and its de-
crease may not be a serious problem. This will
be discussed in another section of this paper.
Of considerable interest is the effect of
increased dilution rate on changes in the macro-
molecular composition of the microbial cells
produced. As dilution rate increases, the rela-
tive content of RNA increases greatly, whereas
the relative content of DNA and protein de-
clines (Kjeldegaard, 1967). Should this be the
situation in the rumen, improvement in cell
yield may be at the expense of microbial
protein quality and quantity, which would be
an obvious disadvantage to the animal. Lastly,
as dilution rate or microbial growth rate in-
creases, microorganisms will shift their fermen-
tation to less efficient pathways and yet may
show a net growth because of lower mainte-
nance requirements. (Hobson, 1965). Regula-
 RUMEN FERMENTATIONTO MAXIMIZEANIMALPRODUCTIVITY
tion of carbon flow into fermentation end
products as a function of growth rate is not
completely understood. In regard to the latter
two considerations, it has been demonstrated
that increasing dilution rate will markedly
influence bacterial enzyme activities (Matin et
al., 1976).
Manipulation o f Fermentation Products and
Growth Efficiency (Cell Yield). Productive effi-
ciency can be improved in the rumen by
exploiting what is known about the microbial
processes and advantageously manipulating the
end products of fermentation (i.e., volatile
fatty acids and microbial cells). Current re-
search interest has centered on attempting to
inhibit rumen methanogenesis by shifting the
reducing equivalents away from CO2 to end
products of more nutritional value, thereby
conserving the estimated 8% loss in feed energy.
That this is a real possibility has been demon-
strated by in vitro and in vivo studies (Van
Nevel et al., 1969; Rufener and Wolin, 1968)
which show H2 accumulation, as a result of
methane inhibition, with a concomitant shift in
fermentation end-products, primarily to propio-
nate. There is, however, some controversy as to
whether the inhibition of methanogenesis
would be an advantageous manipulation with
regard to ruminal fermentation efficiency. This
controversy concerns the hypothesis that meth-
ane formation serves as an energy sink into
which the electrons from all ruminal microorga-
nisms drain, in turn allowing them a higher
yield of ATP, and generating additional ATP
(Hungate, 1966). In effect, this hypothesis
suggests that methanogenesis modulates micro-
bial cell yield by its effect on net ATP yield,
and its inhibition (conservation of feed energy)
will be offset by a decrease in efficiency of net
microbial growth.
In vitro studies concerned with the interac-
tions of ruminal microorganisms, specifically in
combined cultures, have conceptually con-
firmed this hypothesis (Iannotti et al., 1973).
In these studies, co-culturing Ruminococcus
albus and Vibrio succinogenes demonstrated
that an H2-utilizer (V. succinogenes) can alter
the fermentation end products of an H2-pro-
ducer (R. albus) such that more ATP is made
available for growth. The symbiotic association
of Metbanobacterium ruminantium and S-
organism is another example where the removal
of H2 by the methanogen can stimulate the
growth of the S-organism by almost twofold
(Bryant, 1972). The probable interactions of
579
other ruminal microorganisms under methano-
gen dependent and independent conditions
have been described (Wolin, 1975). In essence,
these model systems (no tri-culture studies have
yet been conducted) suggest that successful
competition for electrons by rumen methano-
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gens shifts the fermentation away from ineffi-
cient pathways (i.e., from ethanol, lactate,
formate and perhaps, succinate) to acetate
production, which should make more ATP
available for growth. Related to this discussion
is the observation that some strains of Seleno-
monas ruminantium, although producing only
trace amounts of H2 in pure culture, will
substantially increase H2 production in the
presence of methanogenic bacteria (Scheifinger
et al., 1973).
If Hungate's hypothesis is accepted, there-
fore, shifting ruminal fermentation away from
methane to propionate production should re-
sult in a decrease in molar cell yield because of
less ATP availability. Demeyer and Van Nevel
(1975) varied ruminal fermentation to change
the proportions of methane and propionate
produced, and measured in vitro N1 s and p32
incorporation to estimate microbial cell yield.
Although they were able to effect a wide
variation in the proportions of these end
products, based on their incorporation data,
they concluded that in batch cultures a shift to
a low methane and high propionate type of
fermentation was not associated with a decrease
in microbial growth efficiency. This would be
contrary to Hungate's hypothesis. On the other
hand, some in vivo results (Jackson et al., 1971)
have demonstrated an increased yields of micro-
bial cells with an increase in propionate produc-
tion. These researchers, however, altered the
physical form of the ration by grinding and
pelleting which may have increased rate of
passage (dilution rate) and thereby the specific
growth rate of the ruminal microorganisms. It is
conceivable that under conditions of decreased
methane production (batch culture or constant
dilution rate in a continuous system) increased
availability of succinate and its conversion to
propionate (perhaps providing additional ATP
via anaerobic cytochromeqinked electron trans-
port) could be making up for or complementing
the ATP generated via acetate formation in the
presence of a competitive methanogen. Seleno-
monas ruminantium, an important propionate
producing bacterium in the rumen, is known to
possess a cytochrome b linked electron trans-
port system, and this property is believed to be
580 BERGEN AND YOKOYAMA
related to its higher molar growth yield (62 g
cells/mole glucose) (Hobson and Summers,
1967).
The Role o f Digesta Passage and Degradative
Rates on Substrate Utilization in the Rumen.
Rate of substrate disappearance from the
rumen is a combined function of ruminal
outflow (passage) and the rate of degradation.
Particle size and specific gravity are the two
primary factors that control the potential exit
of a feed particle from the reticulo-rumen
(Balch and Campling, 1965). Coarse, poorly
digestible roughage will be retained in the
rumen for a long period to achieve breakdown
and will be poorly consumed. Grinding of this
roughage will increase the rate of disappearance
from the rumen and hence intake. An increased
rate of disappearance from the rumen (e.g.,
passage) will actually decrease the extent of
roughage digestion (Van Soest, 1975). The
same phenomenon is not readily observed with
more digestible feedstuffs such as processed
grains as the rate of digestion will generally
exceed the rate of outflow (or dilution rate)
from the rumen (Hungate, 1966).
Hungate (1966) detailed the expected extent
of digestion of either soluble, soluble polymer
and insoluble carbohydrate substrates. The rate
of microbial degradation of soluble carbohy-
drates (simple sugars) vastly exceeds the rate of
ruminal outflow (dilution rate) so that very few
simple sugars escape ruminal fermentation (Hun-
gate, 1966). Medium size, polymerized, soluble
carbohydrates are degraded somewhat more
slowly than are simple sugars and more of these
materials will escape ruminal fermentation.
Regardless of rate of degradation, the outflow
of the insoluble structural carbohydrates is
dependent on particle size, and hence ruminal
outflow will not affect ruminal disappearance
of these materials until the proper particle size
is reached (Hungate, 1966).
A treatment similar to the one for ruminal
carbohydrate disappearance used by Hungate
(1966) is depicted for preformed dietary pro-
tein sources (figure 1). The major deviation
from the above considerations for carbohy-
drates is that proteins resistant to degradation
are not usually physically restrained from leav-
ing the rumen. Figure 1 shows that the extent
of ruminal breakdown (or bypass) of proteins is
a function of both outflow and catabolism.
Proteins that are rapidly catabolized do not
escape the rumen, whereas slowly catabolized
proteins such as zein (McDonald, 1954)disap-
pear from the rumen more or less at the
prevailing outflow rate. Although the optimal
NH3-N (and possibly amino acid) levels (Maeng
and Baldwin, 1976a,b; Maeng et al., 1976) that
are necessary for microbial growth are not
precisely known (estimates vary from 5 mg
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NH3-N to 23 mg NH3-N/100 m. rumen liquor
(Satter and Slyter, 1974; Mehrez and Orskov,
1976)) it is clear that if preformed proteins are
the sole N source in diets of ruminants,
excessive bypass will limit microbial growth and
VFA production. This point was amply demon-
strated in steer feeding trials by Schmidt et al.
(1973) with formaldehyde treated soybean
meal.
The Role o f Ciliate Protozoa in Rumen
Fermentation and Host Metabolism. The extent
of the contribution of ciliate protozoa to the
ruminal carbohydrate and nitrogen metabolism
has long been debated. Oxford (1955) argued
that protozoa provide the host with a higher
quality, "animal type" protein source. The
protein quality (NPU), protein digestibility and
lysine content of ruminal protozoa exceeds
values found in ruminal bacteria (Bergen et al.,
1968a,b). Abou Akkada and el Shazely (1964)
found that the average daily gain of faunated
feeder lambs was significantly higher than for
defaunated lambs, although overall the daily
growth rates were quite low (80 to 100 g/day).
Klopfenstein et al. (1966) reported that defau-
nated sheep fed restricted high grain rations had
a lower apparent dry matter and protein diges-
tion and N balance than did faunated lambs.
Concomitantly, the faunated sheep had consist-
ently higher rumen ammonia-N concentrations
(Klopfenstein et al., 1966) a phenomenon also
noted by other workers (Eadie and Hobson,
1962; Males and Purser, 1970).
Rumen ciliate protozoa engulf bacteria and
use these as a source of amino acids and
possible other growth factors (Coleman, 1975).
Large species of protozoa may also engulf and
use plant proteins directly. The overall rate of
bacterial (S. boris) breakdown by mixed proto-
zoa in the rumen was estimated to be 5.2% per
hour (Jarvis, 1968). The high rate of protozoal
engulfment of bacteria may be responsible for
the much lower ruminal bacterial concentration
generally found in faunated animals (Coleman,
1975; Eadie and Hobson, 1962; Males and
Purser, 1970).
Only about one half of the engulfed bacte-
rial N is utilized by the protozoa and the
remaining portion is released back into the
 RUMEN FERMENTATION TO MAXIMIZE ANIMAL PRODUCTIVITY
1.0
i.~. . Spillover
E x~o.kt
c
0 always been commonly assumed that most
Time
Figure 1. Hypothetical d~pp~rance rate of pre-
formed proteins and digesta spillover (outflow) rate
from the rumen. The disappearance from the rumen of
the fast, medium and slow degrading proteins is a
combined function of the specific degradation rates [x
(a-t) where t < a; x = fraction of protein remaining
after time t; the degradation rate is linear during time
0 ~ aI and the ruminant spillover rate (x ---e-kt, where
x is fraction of outflow marker remaining in the
rumen after time t). The three protein disappearance
curves can be described by the combined general
formula x = e -kt (a-t).
rumen (Coleman, 1975). The higher ruminal
NH3-N levels in faunated animals may be a
direct consequence of the poor protozoal utili-
zation of bacterial N and thus represents a
sizeable inefficiency (or an increase in mainte-
nance energy costs) for overall ruminal micro-
bial protein synthesis. This extensive loss of
bacterial amino acid-N and partial conversion
into protozoal protein has not been interpreted
as a major detriment to the productivity of
ruminants as it was felt that the higher protein
quality and digestibility of protozoa would
counterbalance any net N losses (Coleman,
1975). This view may be somewhat misleading
as the Biological Values of mixed bacterial and
protozoal preparations (at least with rats) are
the same (85 vs 82) (Bergen et al., 1968a) and
only the protein digestibilities differ (75 vs 87)
improving the NPU for protozoa by eight units
(Bergen et al., 1968a). In animals fed high
roughage (low net energy (NE)) rations this loss
of microbial N may not be critical as the
animal's protein requirement depends on the
available energy for growth, b u t in high produc-
ing ruminants fed high NE rations this loss of
microbial N may be critical. It should be
emphasized however that protozoa are seldom
found in high numbers in the rumen of animals
fed high grain rations (Coleman, 1975).
From a variety of experimental approaches
bacterial and protozoal protein synthesis (as a
percent of total microbial protein synthesis)
have been estimated to be 80% and 20%
5 81
(Pilgrim et al,, 1970), 70% and 30% (Hungate et
al., 1971) and 50% and 50% (Bucholtz and
Bergen, 1973), respectively. In all the above
studies roughage based rations were used. It had
always been assumed that the protozoal protein
synthesized in the rumen would quantitatively
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pass to the omasum and then be digested in the
abomasum and small intestine. Further it had
bacteria and protozoa leave the rumen with the
fluid-smaU particle phase. If the rumen fluid
concentration of bacteria and protozoa as well
as the rumen fluid dilution rate were known,
the passage of microbial protein to the lower
gut could be determined. Weller and Pilgrim
(1974) compared the actual appearance of
protozoa in the omasum with an estimated
outflow based on dilution rate data. The work
indicated a passage of protozoa to the omasum
only 6 to 29% of that which was estimated
from the dilution rate data. It would appear
that the protozoa undergo some sort of seques-
tration in the rumen and that protozoal turn-
over is therefore slower than the rumen liquor
dilution rate. (Weller and Pilgrim, 1974).
The above data raise the possibility of a
cycling protozoa pool within the rumen with
little net protozoal protein passage to the lower
gastro-intestinal tract. If this were indeed the
case, ruminal protozoa would actually limit
ruminal protein production and depress the
supply of amino acids to the host.
Coda. The two major reactions of the
ruminal fermentation are the degradation of
carbohydrates to V F A as a method of energy
(ATP) generation and the concomitant produc-
tion of microbial cells. The overall scheme of
ruminal activity is depicted in figure 2.
The capacity for microbial growth depends
primarily on the extent of ATP generation.
Other factors such as the dilution rate (growth
rate) can modulate the maintenance needs for
microbial cells. At high microbial growth or
ruminal dilution rates, the ATP needs for
maintenance are quite low, and much of the
ATP is available for cell synthesis, while at low
dilution rates the major portion of available
ATP is needed for maintenance of the microbial
cells.
For optimal ceils synthesis, other precursors
are needed. These include an adequate supply
of NH3-N, carbon skeletons, sulfur, possibly
free amino acids (Maeng et al., 1976) and other
not well identified cofactors. The process of
V F A production from carbohydrates and cellu-
582 BERGEN AND YOKOYAMA
CHO VFA
NPN t /.Gos
\ M
m, xl~ll-13 /"/' / ! "-Cells
! Is=,,.,/
Protein: sCe~176"mA~~ 7
Other
(co) foetors
Figure 2. A flow diagram of the interrelationship
between carbohydrate fermentation (ATP generation)
and microbial cell growth in the rumen.
lar g r o w t h t o a high degree is a c o u p l e d process.
A d e f i c i e n c y o f a n y one o f t h e c o m p o n e n t s in
t h e overall s c h e m e (figure 2) will i m p o s e limits
a n d d e p r e s s t h e o u t p u t ( V F A and m i c r o b i a l
cells) o f t h e r u m i n a l f e r m e n t a t i o n .
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