Kelompok 4 VFA-1

Animal Feed Science and Technology, 43 ( 1993 ) 189-225 189
0377-8401/93/$06.00 © 1993 - Elsevier Science Publishers B.V. All rights reserved
The production and metabolism of volatile fatty

acids by ruminants fed roughages: A review
M.F.J. van Houtert ~

Department of Biochemistry, Microbiology and Nutrition, The University of New England, Armidale,
N.S. IV.. 2351, Australia
(Received 11 August 1992; accepted 8 April 1993)
Abstract
The production, absorption and pathways of utilisation of volatile fatty acids (VFAs) by ruminants
are reviewed. Acetate fulfils a central role in ruminant metabolism, which is interdependent upon
other key nutrients such as other main VFAs, amino acids and glucose. The interaction of these key
nutrients has received little attention, particularly in roughage-fed ruminants. From theoretical con-
siderations, the ratio of acetate to propionate absorbed into circulation appears to affect the energetic
efficiency of utilisation of VFAs. While there is a multitude of conflicting experimental results, it
seems that under practical circumstances, the ratio of acetate to propionate absorbed has little impact
on the energetic efficiency of ruminant production.
Introduction
The energy associated with the phosphoanhydride bonds of adenosine tri-

phosphate (ATP), or similar nucleoside triphosphates, is the major energy
source for microbial cell metabolism. Since complete degradation of substrate
(feed) to carbon dioxide and water is prevented because of the anaerobic
conditions in the rumen, fermentation of substrate is the mechanism by which
ATP is generated. Fermentation is the process of energy-yielding oxidation-
reduction reactions in which both the initial electron donor(s) and final elec-
tron acceptor(s) are organic compounds (Prins, 1977 ). Only a small part of
the energy in plant carbohydrates is transferred to ATP during fermentation,
however. Most of the energy is retained in a number of by-products of fer-
mentation, while some is lost as heat. The volatile fatty acids (VFAs) are the
most important by-products. A range of other by-products (e.g. hydrogen,
ethanol, formate, lactate and succinate) are formed by some microbial spe-
cies, but these are rapidly utilised by other bacterial species with the produc-
tion of VFAs, methane and carbon dioxide. Methane cannot be utilised any
Present address: CSIRO Division of Animal Health, Private Bag, P.O. Armidale, N.S.W. 2350,
Australia.
190 M.F.J. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225
further within the anaerobic rumen and is lost, together with carbon dioxide,
through absorption and respiration and eructation.
Acetate, propionate and butyrate are the key VFAs formed in the rumen
from fermentation. As long ago as 1884, Tappeiner speculated that acetate
and other VFAs could be of some energetic value to the animal. Mall~vre
(1891), with his infusion studies with fasting rabbits, proved that acetate
spared tissue (fat) from oxidation and as such had a nutritive value, albeit
less than that of butyric acid or lactic acid. He did not exclude the possibility,
however, that the net value of the VFAs formed from fermentation of cellu-
lose was nil, owing to increases in the energy cost of digestion of fibrous diets.
Despite this early recognition of the nutritive value of VFAs for herbivores,
the view persisted well into this century that glucose molecules split off from
cellulose, together with microbial cells, constituted the major nutrient source
for the ruminant (Baker, 1942). One school of thought advocated that ru-
minants relied solely on the microbial cells to meet their nutrient require-
ments (Hastings, 1944). However, work at Cambridge around 1940 demon-
strated unequivocally that VFAs constituted the major energy source for the
ruminant (Phillipson and McAnally, 1942; Barcroft et al., 1944). It has since
been estimated that VFAs provide the ruminant with up to 0.7-0.8 of all its
energy requirements (Warner, 1964; Bergman et al., 1965; Annison and Arm-
strong, 1970). In addition to acetate, propionate and butyrate, higher VFAs
were also shown to be produced in the rumen in minor quantities (Elsden,
1946; Elsden and Phillipson, 1948; Annison, 1954).
Selected aspects of the production and metabolism of VFAs and their inter-
action with other major nutrients, which together determine the nutrient
economy of the growing roughage-fed ruminant, are reviewed here. Given the
broad area of relevant research, review of certain aspects had to be omitted.
This includes techniques for measuring VFA production and absorption and
effects of drugs such as antibiotics and ionophores on VFA production and
metabolism.
Fermentation of carbohydrates to volatile fatty acids
Although ruminal fermentation of dietary proteins contributes to the pro-

duction of VFAs, in roughage-fed ruminants acetate, propionate and butyrate
originate mainly from the fermentation of dietary carbohydrates. Plant car-
bohydrates can be classified into three major categories (Van Soest, 1982).
There are the simple sugars which are active in plant intermediary metabo-
lism, the storage reserve carbohydrates such as sucrose, fructans and starch
and the complex structural carbohydrates such as the pectins and hemicellu-
loses and cellulose. The biochemical pathways involved in the fermentation
of dietary carbohydrates to VFAs have been reviewed by Baldwin ( 1965 ),
Walker (1965), Hungate (1966), Leng (1970a, 1973), Prins (1977), De-
M.F.J. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 191
meyer ( 1981 ), Wolin and Miller (1983), Czerkawski (1986) and Bergman
(1990), amongst others. A simplified summary of these pathways is pre-
sented in Fig. 1.
Yields of ATP from substrate fermentation cannot be measured directly,
but must be deduced from known pathways of carbohydrate catabolism. It
has been shown that ATP synthesis in anaerobic microorganisms does not
only proceed via substrate-level phosphorylation reactions, but also involves
electron transfer-linked phosphorylation, at least in some microbial species
(Erfle et al., 1986). Part of this ATP may be used for transport processes of
molecules across cell membranes (Hespell and Bryant, 1979). The yield of
ATP from fermentation of a particular substrate thus varies with microbial
S ch I I C'l"'oso I I H°mlc°"u'°s°I I P °t n U
4, 4, 4, 4,
~l,o~o I I c01'o~io~o I X~,ob,o~e I ~oX~d 1
Glucose I 1 [ Glucose I Xylose[ I~ ~v GalacturoniCacid
Glucose- 6 - P XyluloseP RiboseP

~ ---
~ 6 ~ ~ [ ~ F \ t ..... P [ <~1 Eryth.... P

IDihydroxyaceton~k---'-]
P I "1 Glyceraldehyde
-3~ - ]I~x ~ -~ I FructoseP II~"--I [ TrioseP
I ]
3PGly...... 1~----11,3di-P-Glycerat~ I Aoo'o~oe','~oAF--~[ ~O~o~'r~' ]

2P Glycerate [ ~l Phosphoenolpyruvate
] I Malo.ylCoA j I Cro,onylCoAJ
--I 4' ~'
-'1 AcctylCoA ~- [ ButyrylCoA]
~a~'~'CoA I I Oxa,o
~,,ylCoX,151 s0
~o~ioo,l CoAI I M.lon,lCoA r
--+1~
~1 A~,,IP II
]1
4'
C== . . . . . .il. . . .
Fig. 1. Schematic and simplified diagram showing the pathways of fermentation of carbohy-
drate via pyruvate to the main volatile fatty acids and methane. Adapted from Leng (1973 )
and Czerkawski (1986).
192 M.F.J. van Houtert /Animal Feed Science and Technology 43 (1993) 189-225
species. Although there is general agreement on the yield of ATP per mole of
hexose fermented to acetate, this does not appear to be the case for the yields
of ATP in the fermentation of hexose units to propionate or butyrate (Hard-
son and McAllan, 1980). Tamminga ( 1979 ) suggested yields of ATP per mole
of hexose fermented of between 3.5 and 5.3, with the final ratios of VFAs
produced having relatively little effect on ATP yield. Estimates of the ATP
yield of fermentation of feed proteins and lipids are considerably lower than
for carbohydrates (Tamminga, 1978). Equations ( 1 ) - (5) represent simpli-
fied net stoichiometries of the major reactions which are believed to approach
in vivo carbohydrate catabolism (adapted from Wolin, 1960, 1975; Hungate,
1966; Leng, 1973; Tamminga, 1979; Czerkawski, 1986 ). As pointed out above,
the actual ATP yield for reaction (3) in the rumen is equivocal.
C6 H 1206 + 2NAD + + 2ADP
=2CH3COCOOH+2[H] +2NADH+2ATP (1)
and subsequently
2CH3 COCOOH + 2H2 O + 2ADP
= 2CH3 COOH + 2CO2 -I-2H2 + 2ATP (2)
2CH3 COCOOH + 4 [H ] + 4NADH + 2ADP

= 2CH3CH2 COOH + 2 H 2 0 + 4NAD+ + 2ATP (3)
2CH3 COCOOH + 2 [ H ] + 2NADH + ADP

=CH3CH2CH2COOH+2H2-t-2CO2 + 2 N A D + + A T P (4)
CO2 + 4H2 + ADP = CH 4 + 2H20 + ATP (5)
Fermentation of complex carbohydrates to pyruvate
Cellulose and hemicelluloses

These are the most abundant of the plant carbohydrates. While hemicellu-
loses are slightly more resistant to microbial degradation than cellulose
(Tamminga, 1979 ), both are fermented slowly in the rumen. Breakdown of
cellulose commences under the influence of cellulase complexes, which are
produced as extracellular enzymes by ruminal bacteria such as Fibrobacter
succinogenes (previously known as Bacteroides succinogenes ), Ruminococcus
flavefaciens and Ruminococcus albus and to a lesser extent, Butyrivibriofibri-
solvens and Eubacterium cellulosolvens (Hungate, 1966; Schwartz and
Gilchrist, 1975). Certain fungi (e.g. Micromonospora ruminantium) and
protozoa (e.g. species of the genera Diplodinium, Entodinium, Eudiplodi-
M.F.,L van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 193
nium and Polyplastron) also appear to possess ceUulolytic activity (Prins,

1977; Wolin and Miller, 1983). Cellulose is degraded via a number of steps
to oligosaccharides, then to cellobiose and finally glucose. Hemicelluloses are
broken down enzymatically by ruminal bacteria, such as Butyrivibriofibrisol-
vens, to xylose and other pentoses and via a further complex series of conver-
sions to fructoses and trioses (Baldwin, 1965; Leng, 1970a). Cell-free ex-
tracts of a number of protozoal species have been shown to degrade
hemicelluloses (Hungate, 1966).
Pectins
Pectins are an important group of structural carbohydrates found in the
plant cell wall. They are fermented much more rapidly than cellulose and
hemicelluloses (Tamminga, 1979), with the involvement of extracellular
pectolytic enzymes excreted by bacterial species such as Fibrobacter succino-
genes, Prevotella ruminocola (previously known as Bacteroides ruminocola ),
Butyrivibrio fibrisolvens, Eubacterium cellulosolvens, Lachnospira multiparus
and Succinovibrio dextrinosolvens (Prins, 1977; Harfoot, 1978). Pectins are
broken down to pectic acid and galacturonic acid and subsequently via xylose
to fructoses and trioses (Prins, 1977 ).
Storage carbohydrates
Starches (amylose and amylopectin) and other storage carbohydrates do
not abound in fibrous roughages. Those ingested by the ruminant are in gen-
eral broken down rapidly under the influence of amylases and other carboh-
ydrases to maltose and subsequently to glucose units. Starches derived from
grains and other concentrated feed sources may differ significantly in their
rate of microbial degradation. Corn starch and sorghum starch are fermented
less rapidly than barley starch (Waldo, 1973 ). The most important ruminal
starch degraders are Bacteroides amylophilus, Streptococcus bovis and Succi-
nomonas amylolytica (Prins, 1977 ). The protozoa also play a prominent role
in storage carbohydrate catabolism by ingesting starch and taking up soluble
sugars (Schwartz and Gilchrist, 1975 ). These carbohydrates are stored within
the protozoa and slowly metabolised, thereby reducing their availability to
other microbial species.
Fermentation to pyruvate
The simple stoichiometries outlined in Eqs. (1)-(5 ) predict the actual
yields of ATP and VFAs and other breakdown products with only a limited
accuracy. This is because hexose monomers are the dominant, but not the
only intermediate monomers produced as a result of fermentation of carbo-
hydrates and proteins, and a range of other unknown factors (Tamminga,
1978 ). All the carbohydrate monomers which are formed are catabolised to
pyruvate. The glycolytic Embden-Meyerhof-Parnas pathway has been shown
194 M.F.£ van Houtert /Animal Feed Science and Technology 43 (1993) 189-225
to constitute the major route of further hexose catabolism by the microorga-

nisms in the rumen (Baldwin, 1965 ). Although alternative pathways for the
conversion ofhexoses to pyruvate may occur, such as those involving 6-phos-
phogluconate, these result in only 1 mol ATP being generated per mole of
glucose fermented and do not seem to be of significance within the rumen
(Prins, 1977). For entry into the Embden-Meyerhof-Parnas pathway, glu-
cose and fructose monomers are phosphorylated at the initial expense of 2
mol ATP mol-~ hexose. Involvement of phosphorylases in the cleavage of
disaccharides into monomers requires only 1 mol ATP (Tamminga, 1979).
Phosphorylated hexoses are then split over a number of steps to yield glycer-
aldehyde-3-phosphate, whereas triose-phosphate monomers are directly con-
verted to glyceraldehyde-3-phosphate. The glyceraldehyde-3-phosphate is then
converted, again via a number of steps, to phosphoenolpyruvate and finally
pyruvate. The latter process yields 4 mol ATP per mole of phosphorylated
hexose, resulting in a net yield of 2 mol ATP per mole ofhexose fermented to
pyruvate or 2.5 mol if phosphorylases are involved (Tamminga, 1979).
Fermentation of pyruvate to volatile fatty acids
The central role of pyruvate in rumen fermentation continues, with the

conversion of pyruvate to acetate, propionate and butyrate via a number of
major pathways (see Fig. 1 ), again under the influence of enzymes secreted
by a multitude of microbial species. It should be emphasised, however, that
pyruvate itself is not a major end product of metabolism of any of the impor-
tant rumen bacteria. The three important cellulose degraders (see earlier)
produce end products such as succinate, lactate, hydrogen and formate as well
as VFAs, in which pyruvate is merely a common intermediate. The non-VFA
end products are then used as substrates for other bacterial species which con-
vert these to VFAs a n d / o r methane as their own end products.
Fermentation of feed to the various VFAs is determined to a large extent
by conditions favouring or diminishing the activities of particular groups of
ruminal microorganisms. The availability and nature of the substrate, the
presence of electron donors and acceptors and microbial interactions are some
of the many factors which influence the pathways of conversion of pyruvate
to VFAs (Sutherland, 1977; Tamminga, 1979). For instance, the methano-
genic species Methanobacterium ruminantium and Methanobacterium mo-
bile play key roles in rumen fermentation by maintaining low concentrations
of hydrogen in the rumen milieu. This facilitates 'normal' metabolism of the
other bacterial species (Schwartz and Gilchrist, 1975; Wolin and Miller, 1983;
Erfle et al., 1986 ). Thus the highly interactive ecology ofrumen fermentation
becomes apparent (Wolin, 1975; Schwartz and Gilchrist, 1975; Prins, 1977 ).
There are many aspects of these interactions which have not been studied in
detail. Nevertheless, an outline of all known factors influencing pathways of
M.F.J. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 19 5
VFA production by the various rumen microbes is beyond the scope of this
review. Only some of the more important theories related to the regulation of
the production of acetate, propionate and butyrate from carbohydrates will
be discussed below.
Fermentation to acetate
The production of acetate from pyruvate proceeds under the influence of a
number of different enzyme sequences, each occurring in different bacterial
species. A prominent pathway for the conversion of pyruvate to acetate in the
rumen is the cleavage of pyruvate to acetyl-phosphate and formate under the
influence of pyruvate-formate lyase, a reaction previously known as the phos-
phoroclastic reaction (Leng, 1970a, 1973).
Important factors in the regulation of fermentation of carbohydrates to the
different VFAs are ruminal pH (Peters et al., 1989), the requirement for an
electron acceptor (NAD + ), the generation of reduced nicotinamide adenine
dinucleotide (NADH) in the breakdown of carbohydrate to pyruvate (see
Eq. (1)) and also substrate pressure and the related ruminal dilution rate
(Wolin, 1975 ). Low rates of feed intake and feed fermentation with a pH in
the rumen of 6.5-7 are common in ruminants fed roughage-based diets. This
is reflected in a low rate of carbon flow from carbohydrate to pyruvate which
assists in maintaining a relatively high ratio of NAD +/NADH in the rumen
ecosystem. This reduces the flow of carbon through electron-accepting path-
ways such as propionate production. As a consequence of the nature of the
substrate (mainly structural carbohydrates), the low rate of rumen turnover
and the ratio of electron donors and acceptors, the bacterial species convert-
ing carbohydrate into acetate proliferate at the expense of propionate fermen-
ters in ruminants fed roughages (Sutherland, 1977 ).
Fermentation to propionate
The formation of propionate from carbohydrate proceeds via two major
pathways in the rumen (Baldwin, 1965; Leng, 1973; Wolin, 1975; Bergman,
1990). After the catabolism of complex carbohydrates to pyruvate, pyruvate
is condensed with carbon dioxide to form oxaloacetate or malate and then
converted via fumarate to succinate (Baldwin, 1965), a major end product
of a range of bacterial species as outlined earlier. Succinate is then used as a
substrate by other microbial species, which convert it to propionate over a
number of steps involving decarboxylation (Baldwin, 1965; Wolin, 1975 ).
This pathway, known as the succinate, dicarboxylic acid or randomising
pathway, appears to dominate propionate production in the rumen of ani-
mals fed roughages. A second pathway of production of propionate involves
the conversion ofpyruvate to propionate via lactate and acrylate, the so-called
acrylate, glyoxalate, direct reductive or non-randomising pathway. This path-
way takes prominence only in the rumen receiving high levels of concentrates
(mainly grains but also beans and oil-seed by-products), where lactate may
be produced in appreciable or even deleterious amounts (Schwartz and
Gilchrist, 1975; Tamminga, 1979).
As outlined in Eq. (3), production of propionate from carbohydrate re-
quires the donation of electrons from NADH. The feeding of ruminants on
forage- or concentrate-based diets results in rates of feed intake and feed fer-
mentation higher than those achieved when roughages are fed. A high rate of
flow of carbohydrate carbon through the glycolytic pathway to pyruvate leads
to a higher ratio of NADH relative to NAD ÷ and to high concentrations of H
ions (lowered pH). Both of these factors and the higher rates of rumen dilu-
tion favour bacterial species with enzymes converting carbohydrate to pro-
pionate via the succinate pathway (Sutherland, 1977 ).
Fermentation to butyrate
Butyrate is quantitatively the least important of the three major VFAs in
rumen fluid of animals fed roughages, and usually contributes only 0.05-0.10
on a molar basis to the total VFA pool. Bergman et al. (1965), Leng and
Leonard (1965) and Leng and Brett (1966) showed that there is extensive
interconversion of carbon between the acetate and butyrate pools in the ru-
men. At least half the butyrate was derived from acetate, while proportion-
ately 0.1-0.2 of the acetate was derived from butyrate in the sheep used in
these two studies. The conversion ofbutyrate to acetate results in the net gen-
eration of ATP.
Two main mechanisms for the synthesis of butyrate have been proposed
(Baldwin, 1965 ). Reversal of the classical fl-oxidation of fatty acids (Lehn-
inger, 1982) appears to be the main mechanism for condensation of 2 mol
acetate into 1 mol butyrate. Two moles of acetate are activated to acetyl-CoA,
which is then condensed to 1 mol acetoacetyl-CoA. This is then converted in
a number of steps to butyrate which requires electron donation from NADH
(Leng, 1973 ). This pathway of butyrate formation has been shown to occur
in at least one strain of Butyrivibriofibrisolvens (Miller and Jenesel, 1979)
and generates 1 mol ATP. Carboxylation of acetyl-CoA to malonyl-CoA and
the subsequent reaction ofmalonyl-CoA with acetyl-CoA to yield acetoacetyl-
CoA (analogous to the initial steps of long chain fatty acid synthesis; Bald-
win, 1965; Lehninger, 1982) is another potential pathway leading to butyrate
synthesis, without the net gain ofATP (Leng, 1973 ).
Methanogenesis
As indicated earlier, methanogenesis by M. ruminantium and M. mobile
plays an important role in rumen fermentation. Carbon dioxide, hydrogen
gas and formate are the three major precursors in the production of methane.
Methane itself cannot be utilised any further in the reduced environment of
the rumen and is disposed of through eructation and respiration. This consti-
tutes an energy loss to the animal which has been estimated to range from
0.06 to 0.17 of the digestible energy available to the animal (Orskov, 1975 ).
As outlined in Eq. (3), formation of propionate also requires hydrogen.
Hence, production of methane and propionate both compete for hydrogen
with other reducing reactions such as production ofbutyrate and synthesis of
some amino acids. Methane-producing bacteria are slow growing and thus
proliferate especially under conditions of slow rumen particle dilution rate as
found in ruminants fed roughages or molasses as the basal diet (Preston,
1972).
Considerable research efforts have gone into attempts to reduce the energy
loss to the animal in the form of methane (e.g. Czerkawski et al., 1966, 1975;
Demeyer and Henderickx, 1967; Van Nevel et al., 1969, 1971; Henderson,
1973; Wainman et al., 1982). Because of the generally inverse relationship
between methane production and propionate production in the rumen
(Orskov et al., 1968; Van Nevel et al., 1969; Peters et al., 1989), suppression
of methane production is accompanied by a reduction in the non-gluco-
genic:glucogenic ratio (molar proportion of (acetic acid+2 butyric
acid + valeric acid ) / (propionic acid + valeric acid ); IDrskov, 1975 ) of VFAs
produced in the rumen. Also, inhibition of methane production may result in
a reduction in the net microbial cell yield associated with fermentation of
feed. These two factors, and also possible adaptations of microbial species to
methane inhibitors, in part explain why they often do not improve animal
productivity in the long term (IDrskov, 1975 ).
Efficiency of microbial growth
Factors determining the efficiency of microbial growth are of great impor-

tance for the nutrient economy of the host ruminant. Microbial cell DM mass
(g) produced per mole of ATP resulting from substrate fermentation ( yATP;
Bauchop and Elsden, 1960) is one way of expressing the efficiency of micro-
bial growth. Because of technical difficulties in estimating yAXP,efficiency of
microbial protein synthesis is also expressed commonly as yield of microbial
crude protein or microbial organic matter per kilogram of organic matter fer-
mented (Tamminga, 1978). While yAa'P may vary from 4 to as much as 21
(Harrison and McAllan, 1980), depending on a large number of factors, val-
ues in the range of 10-12 seem to prevail. Values for microbial protein syn-
thesis per kilogram of organic matter apparently fermented in the rumen
ranged from 63 to 307 g with a mean of approximately 170-190 g (Smith,
1979; Stern and Hoover, 1979).
Some of the factors affecting efficiency of microbial growth and its inter-
relationships with VFA production are summarised briefly below. The pro-
posed relationship between microbial growth efficiency and the proportion of
198 M.FJ. van Houtert /Animal Feed Science and Technology 43 (1993) 189-225
% of rumen DE
80
70
60
VFA
50
40
Cells DM 1 / _~I
30
J
/
20 CH4 ~
10
J
j~ l C) 8 ............... e ' " ~ .... " .I
/ Heat
0 I I I I I F I I I I i
0 2 4 6 8 10 12 1~- 16 18 20 22 24
Yatp
Fig. 2. The relationship between yATPand the proportion of rumen digestible energy (DE)
entering microbialcells,VFA,methane and heat (after Leng, 1982a).
fermentable organic matter entering microbial cells, VFA, methane and heat
is shown in Fig. 2 (after Leng, 1982a).
Microbialfactors
For maximal efficiency of microbial cell synthesis, it is desirable to stimu-

late steady and vigorous growth (Smith, 1979 ). The specific growth rate and
maintenance requirements for ATP of the microbial population are a major
influence on yATP (Stouthamer and Bettenhausen, 1973; Hespell and Bryant,
1979; Erfle et al., 1986). At low specific growth rates (equivalent to low di-
lution rates under steady state conditions), a large proportion of ATP which
is generated is required for cell maintenance. Two factors are of major influ-
ence on maintenance requirements for microbes (Hespell and Bryant, 1979):
the efficiency of phosphorylation, i.e. the actual synthesis of ATP from sub-
strate catabolism, and the degree of energetic uncoupling, when the potential
for substrate catabolism and ATP generation is higher than the ability to use
ATP for anabolic purposes.
The nature of the precursor molecules which are used for macromolecule
synthesis, the mechanisms of transport of precursors into microbial cells and
the final chemical composition of the microbial cells all appear to play a role
in determining yATP. The ATP requirements for microbial macromolecule
synthesis are higher when glucose and inorganic salts are used as precursors
than when glucose and other preformed monomers such as amino acids, nu-
cleic acid bases and fatty acids are used (Hespell and Bryant, 1979 ).
The extent of predation by protozoa on bacteria and the subsequent pref-

erential retention of protozoa in the rumen are factors known to reduce net
efficiency of microbial protein synthesis. These areas, as well as the effects of
protozoa on VFA production in the tureen, have been the subject of much
research work, a survey of which is beyond the scope of this review (see Becker
and Everett, 1930; Baker, 1942; Hungate, 1966, 1975; Weller and Pilgrim,
1974; Coleman, 1979, 1988; Leng, 1982a,b; Bird, 1985, 1991; Veira, 1986;
Ryle and Orskov, 1987; Jouany et al., 1988; Bonhomme, 1990; Ushida et al.,
1991).
Fermentable substrate
The nature, quantity and regularity in supply of fermentable substrate are

major determinants of yAXP (Smith, 1979; Stern and Hoover, 1979; Harrison
and McAllan, 1980). The major nitrogen (N) source for microbial protein
synthesis is ammonia. A concentration of between 3 and 5 mmol l-~ (raM)
has been proposed as being sufficient to maximise microbial cell growth (Sat-
ter and Slyter, 1974; Okorie et al., 1977 ). Others however, have reported that
concentrations of 10-20 mM were required to optimise microbial cell yield,
fermentative digestion and/or feed intake (Allan and Miller, 1972; Miller,
1973; Mehrez et al., 1977; Leng and Nolan, 1984; Boniface et al., 1986; Per-
dok and Leng, 1989). It may be that increased rates of fermentation which
were associated with concentrations of rumen ammonia well above 3-5 mM
are due to indirect effects of ammonia, such as increases in rumen pH via the
formation of ammonium bicarbonate (Hespell, 1979; Hespell and Bryant,
1979).
Synthesis of bacterial amino acids from ammonia proceeds via two major
pathways: the glutamate dehydrogenase pathway and the glutamine synthe-
tase/glutamate synthase system. The latter pathway requires the input of ATP
and operates under low concentrations of ammonia in the rumen ( 1 mM or
less). The glutamate dehydrogenase pathway does not require ATP and pre-
dominates when ammonia concentrations are much higher than 1 mM (Hes-
pell, 1984). Increases in yATPhave been reported under high ammonia con-
centrations in vitro, as opposed to more limiting ammonia concentrations
(Hespell, 1984; Leng and Nolan, 1984; Mackie and White, 1990 ). At any one
time it seems likely, however, that both pathways operate in addition to a
number of other ammonia-assimilating pathways. The various microorga-
nisms appear to have specific preferences for these separate pathways (Hes-
pell, 1984). Various transaminases ensure the synthesis of a range of amino
acids from glutamate and glutamine and other products of initial ammonia
assimilation.
Although ammonia provides proportionately 0.4-1.0 of the total N re-
quirements for rumen microbes (Stern and Hoover, 1979), peptides consti-
tute another potential source of N for microbial protein synthesis (Nolan and
Leng, 1972; Argyle and Baldwin, 1989 ). While requirements for peptides ap-
pear to be significant for starch degrading bacterial species, cellulolytic spe-
cies appear to have a minimal requirement in this respect (Maeng et al., 1989;
Leng, 1990). The incorporation of preformed amino acids plays a limited
role in microbial protein synthesis, perhaps owing to the low activity of trans-
port mechanisms for free amino acids into the microbial cell (Russell et al.,
1990). Significant increases in ygrP have been reported, however, for rumen
microbes grown in media containing a proportion of protein rather than non-
protein N only (Maeng et al., 1976; Stern and Hoover, 1979; Maeng et al.,
1989 ). It has been suggested that under practical feeding conditions, supply
of methionine, cysteine and phenylalanine may limit the growth of some mi-
crobial species (Smith, 1979; Hespell, 1984). Higher and branched chain fatty
acids are growth requirements or stimulants for rumen microbes which are
derived from protein catabolism in the rumen (Mackie and White, 1990).
Hence, diets low in available true protein may limit the efficiency of micro-
bial growth (Russell et al., 1990).
The nature and degradability of the carbohydrate source is also of major
influence on yATP. Different microbial species have distinct preferences for
their carbohydrate source. Maximum efficiency of incorporation of dietary N
into microbial protein will not be obtained unless the availability of ferment-
able carbohydrate matches the synthetic rate of rumen microbes. Similarly,
for maximum fermentation from a given energy supply, the rate of incorpo-
ration of nitrogenous sources into microbial cells must keep pace with the rate
of fermentation (Smith, 1979 ). A balance between N and energy supply must
be found in order to optimise microbial cell production. If either does not
keep pace, microbial cell synthesis will be depressed (lower yA+P), which in
turn may reduce feed digestibility and therefore potentially feed intake, as
well as VFA production.
Inadequate availabilities ofmacronutrient mineral elements (calcium (Ca),
phosphorus (P), sulphur (S), potassium (K), sodium (Na), chlorine (C1)
and magnesium (Mg)), and a range of micronutrient elements, may limit the
rate of microbial protein synthesis (Durand and Kawashima, 1980; Mackie
and Therion, 1984). Of particular importance here is the availability of P and
S. Microbial protein synthesis requires a continuous supply of P for nucleic
acid formation and S for synthesis of sulphur amino acids. To ensure that
availability of S does not limit microbial protein synthesis, an optimum die-
tary ratio of 1 g S : 10-15 g N has been suggested (Kandylis, 1984). Differ-
ences in the relative availabilities of N and S in the rumen must also be taken
into account (Harrison and McAllan, 1980).
Physical-chemicalfactors
The major physical-chemical factors affecting the efficiency of microbial
protein synthesis are rumen dilution rate, osmotic pressure, pH, buffering ca-
M.FJ. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 201
pacity and oxidation-reduction potential. Increases in liquid dilution rate

generally result in an increased efficiency of microbial protein synthesis
(Harrison et al., 1975 ), and are associated with a reduction in the proportion
of nutrients required for maintenance functions (HespeU and Bryant, 1979).
Liquid dilution rate is determined by the level of feed intake and the compo-
sition of the diet (Bull et al., 1979). Other important regulatory factors in-
clude hormonal and neural mechanisms and the genetic make-up and cli-
matic environment of the animal (Owens and Goetsch, 1986). High liquid
dilution rates are usually obtained with forage-based diets, whereas low liquid
dilution rates are found in grain-fed ruminants (Harrison and McAllan, 1980).
The opposite may be the case for particle dilution rate and the particle-asso-
ciated microbial mass (Bull et al., 1979). In vitro studies have demonstrated
that increases in liquid dilution rate are associated with increases in yAVP
when glucose or starch was used as the energy source. When cellulose was the
substrate, however, yAVP was reduced with increasing liquid dilution rate
(Maeng et al., 1989). This effect is more pronounced when N is supplied
proportionately as 0.75 urea and 0.25 casein, as opposed to the total amount
of N being supplied as urea.
A high osmotic pressure (400 mOsmol kg -~ ) has been demonstrated to
impair cellulolysis and reduce feed intake (Ternouth and Beattie, 1971; Ber-
gen, 1972; Durand and Kawashima, 1980; Mackie and Therion, 1984). The
nature of the diet (in particular mineral intake) and water intake are some of
the factors determining osmotic pressure in the rumen. Also, pH and buffer-
ing capacity are important determinants of microbial growth efficiency (Dur-
and and Kawashima, 1980). A ruminal pH of less than 6.1 has been shown
to inhibit cellulolysis (Mould and Orskov, 1983; Mould et al., 1983), while
the optimum pH range for cellulolysis is between 6.7 and 7.0 and for micro-
bial protein synthesis above 5.7 (Stewart, 1977; Durand and Kawashima,
1980).
The presence or absence of electron donors and electron acceptors (reduc-
tion-oxidation potential) is important in the regulation of the composition
and amount of substrate which is fermented in the rumen via different cata-
bolic pathways. Methanogenesis is an important hydrogen (electron) sink and
therefore determines the reduction-oxidation potential of the rumen milieu.
Methanogenesis thus appears to be a significant factor in the rate and effi-
ciency of substrate fermentation and microbial cell synthesis (Russell et al.,
1990).
Absorption of volatile fatty acids from the rumen
Early in this century, the rumen epithelium was believed to be impermea-

ble, allowing no direct exchange of metabolites between the rumen ecosystem
and the host animal. The research by Phillipson and McAnally ( 1942 ), Bar-
croft et al. (1944) and McAnally (1944), however, showed unequivocally

that VFAs were absorbed directly from the rumen into the blood. As a result,
the histology of rumen epithelium was studied in detail. The rumen wall con-
sists of smooth muscle tissue lined on the lumen side with stratified squamous
and non-glandular epithelium, which has a highly increased surface area due
to many folds and dense papillae (Dobson et al., 1956). Numerous mito-
chondria in epithelial cells testify to their relatively high metabolic rate.
The negative logarithm of the dissociation constant (PKa) of acetic acid is
4.76 and of propionic acid 4.87. At a pH of between 6 and 7 (the pH of the
rumen contents most commonly found in roughage-fed ruminants), propor-
tionately about 0.87-0.97 of acetic, propionic and also butyric acid present
in the rumen is in the dissociated form (Lehninger, 1982). Since the rate of
absorption of VFAs has been shown to increase with a decrease in rumen pH,
this indicates greater absorption of the undissociated than the dissociated acid
(Danielli et al., 1946 ). These workers and others (e.g. Barcroft et al., 1944)
reported that the rate of absorption of undissociated VFAs increased with
increasing chain length, whereas the reverse was the case for dissociated acids.
It has since been shown that a range of factors influences the relative rates of
absorption of the three major VFAs. These include the total concentration
and the ratios of the VFAs which are present, the pH in the rumen and the
rate of blood flow through the rumen epithelium (Annison et al., 1957; An-
nison and Lewis, 1959; Stevens, 1970; Weigand et al., 1972b).
Stevens (1970) proposed a model for the absorption of VFAs in which the
cell membrane separating the rumen wall from the lumen is permeable to
both forms of the VFAs, whereas the membrane separating the rumen wall
from the blood vessels is permeable to undissociated VFAs only. It is hypoth-
esised that dissociated VFAs which are absorbed into the rumen wall are hy-
drogenated, with carbonic acid acting as the intracellular hydrogen donor.
The excess of intracellular bicarbonate formed as a result of hydrogen dona-
tion by carbonic acid is then excreted back into the lumen (Stevens, 1970;
Stevens et al., 1980). These authors concluded that passive diffusion is the
major mechanism by which VFAs are absorbed from the rumen into the blood.
As most of the produced VFAs are absorbed into the blood stream through
the walls of the rumen and omasum, only a small proportion of the VFAs
enters the abomasum with the digesta flow (Phillipson and McAnally, 1942;
Leng, 1973; Bergman, 1990). Proportional outflow of propionate from the
rumen with the digesta flow at levels as high as 0.3-0.6 of the total ruminal
propionate production have been documented, however, for steers fed diets
rich in corn (Peters et al., 1990a,b). Under these conditions, the removal of
propionate from the digestive tract prior to the duodenum was proportion-
ately 0.97-0.99 of the total ruminal propionate production, indicating sub-
stantial absorption of VFAs into the bloodstream from the omasum and
abomasum.
Metabolism of volatile fatty acids in the rumen wall
The concentrations of the three major VFAs in blood are low in comparison
with their concentrations in the rumen. When taking into consideration their
continuous production in and absorption from the rumen, the low blood con-
centration is indicative of the rapid utilisation of VFAs by the animal (Bar-
croft et al., 1944). Under normal feeding conditions, acetic acid has been
found to constitute the major VFA in blood. The concentration ofbutyrate in
blood in particular is much lower than what would be expected on the basis
of its contribution to the total pool of VFAs in the rumen (Annison et al.,
1957). Subsequent studies of this phenomenon have demonstrated an active
role of rumen epithelium in metabolising VFAs during their transport from
the rumen to the blood (see below). Typically, about 0.9 of the butyrate which
is produced in the rumen of sheep is metabolised by the epithelium of the
rumen and omasum. These figures are estimated at typically 0.5 for propion-
ate and 0.3 for acetate (Bergman and Wolff, 1971; Bergman, 1975, 1990).
Metabolism of acetate
Relatively little acetate is metabolised in the rumen epithelium (Penning-

ton and Pfander, 1957; Cook and Miller, 1965 ), mainly because of a low ac-
tivity of the enzyme acetyl-CoA synthetase in this tissue (Ash and Baird,
1973 ). Leng and West (1969) estimated that proportionately 0.01-0.04 of
the acetate is directly converted to ketone bodies. A small proportion of ace-
tate is oxidised to carbon dioxide, but the major part passes through the epi-
thelium unaltered and is absorbed into the portal blood stream.
Metabolism of propionate
On the basis of early in vitro and in vivo research, it was suggested that at
least half of the propionate was metabolised to lactate in the rumen epithe-
lium, prior to conversion into glucose in the liver (Pennington and Suther-
land, 1956b; Leng et al., 1967; Bergman and Wolff, 1971 ). Subsequent re-
search with cattle (Weigand et al., 1972a) has demonstrated that
proportionately less than 0.05 of all propionate produced in the rumen is con-
verted to lactate in rumen epithelium. Butyrate has been shown to strongly
inhibit the metabolism ofpropionate by rumen epithelium in vitro, as a result
of the inhibition in activity of propionyl-CoA synthetase (Ash and Baird,
1973 ). Nevertheless, in vivo studies by Weekes and Webster (1975) found
that 0.3-0.5 of all propionate is metabolised, but without significant produc-
tion of lactate and therefore presumably as a result of oxidation to carbon
dioxide. The formation of other metabolites, such as glutamate, cannot be
ruled out, however. Weekes and Webster (1975 ) concluded that the forma-
tion of lactate by the gastrointestinal tract probably occurs mostly from glu-
cose in glycolysis. It appears that the extent of metabolism of propionate in
rumen epithelium is less in cattle than in sheep. This may be due to differ-
ences in activity ofpropionyl-CoA synthetase in the rumen epithelium of these
two species (Elliot, 1980). In both species the liver appears to be the more
important site for propionate metabolism than the rumen epithelium.
Metabolism of butyrate
Butyrate is metabolised by the epithelium of the rumen and omasum to the
ketone bodies acetoacetate and fl-hydroxybutyrate. Although acetone is in-
cluded in the group of ketone bodies, it is most probably formed from non-
enzymatic decarboxylation of acetoacetate and contributes only a minor frac-
tion to the total flux of ketone bodies (McGarry and Foster, 1980). The ru-
men epithelium is considered to be the most active ketogenic organ in fed and
non-pregnant sheep (Pennington, 1952; Pennington and Sutherland, 1956a;
Katz and Bergman, 1969; Leng and West, 1969; Varnam et al., 1978 ). This is
in contrast to monogastric animals, where the liver is the only important ke-
togenic organ. Acetoacetate is the main ketone body formed from butyrate by
rumen epithelium of sheep in in vitro incubation trials (Pennington, 1952 ).
Since fl-hydroxybutyrate is the main circulating ketone body in fed ruminants
(Leng, 1965; Baird et al., 1968; Katz and Bergman, 1969; Varnam et al.,
1978), Leng and West (1969) speculated that acetoacetate is reduced to fl-
hydroxybutyrate upon reaching the liver. Subsequent studies on the activity
offl-hydroxybutyrate dehydrogenase in rumen epithelium and liver by Koun-
dakjian and Snoswell (1970) refuted this. They suggested that the capacity
of the liver to convert acetoacetate to fl-hydroxybutyrate is very limited and
that the mitochondria in rumen epithelium are the likely site for this conver-
sion. Subsequent research has confirmed this conclusion (Weigand et al.,
1972b; Emmanuel et al., 1982; Giesecke et al., 1985 ). Two major pathways
have been shown to be active in the conversion of butyrate to fl-hydroxybu-
tyrate. These both proceed with the activation of butyrate to butyryl-CoA.
Activation of VFAs under the influence of acyl-CoA synthetases is an impor-
tant regulating mechanism determining the rate of uptake and utilisation of
VFAs by tissues (Stangassinger and Giesecke, 1986 ). The activated butyryl-
CoA is converted to crotonyl-CoA and L-fl-hydroxybutyryl-CoA (Emmanuel
et al., 1982 ). This is then converted to acetoacetyl-CoA, acetoacetate and fi-
nally fl-hydroxybutyrate (pathway 1 ) or directly converted to D-fl-hydroxy-
butyryl-CoA which is then converted to fl-hydroxybutyrate (pathway 2 ).
Leng and West (1969) showed that in fed sheep, between 0.78 and 0.94 of
all circulating fl-hydroxybutyrate arose from rumen butyrate, with only small
contributions from acetate or non-esterified long-chain fatty acids (LCFAs).
Only 0.15 of all butyrate which was produced in the rumen was converted
into ketone bodies in their studies. As much as 0.34-0.64 of all butyrate pro-
duced in the rumen appeared to be oxidised to carbon dioxide in fed sheep
(Annison et al., 1967). The estimates of oxidation are inflated, however, ow-
ing to the extensive interconversion of carbon between acetate and butyrate
within the rumen (cf. Pennington and Pfander, 1957; Leng and Leonard, 1965;
Bergman et al., 1965; Leng and Brett, 1966 ). Weigand et al. (1972b) reported
that 0.33-0.78 of all butyrate was converted to ketone bodies, mostly fl-hy-
droxybutyrate. Work by these authors and more recent studies by Beck et al.
(1984) indicate that the proportion of butyrate which is either oxidised to
carbon dioxide or converted to ketone bodies depends on the concentration
of the substrate and the pH in the rumen. The metabolism of butyrate to ke-
tone bodies has been proposed as a mechanism to provide the rumen epithe-
lium with most of its energy requirements (Hird and Weidemann, 1964; Bush
and Milligan, 1971 ). Metabolism of butyrate to ketone bodies in rumen epi-
thelium may be beneficial on other grounds also (Giesecke et al., 1985 ). Bu-
tyrate can inhibit cell division and conversion to fl-hydroxybutyrate can be
seen as detoxification. Ketone bodies are not utilised to any extent by the
ruminant liver and therefore they provide extra-hepatic tissues with an addi-
tional energy source. Also, p-hydroxybutyrate may play an important role in
metabolic regulation and control of feed intake in ruminants.
Gluconeogenesis and other pathways of metabolism of volatile fatty acids in

the liver
All metabolites which are absorbed into the blood from the digestive tract
are first transported via the portal vein to the liver (Annison et al., 1957;
Bergman, 1975 ). The liver is involved in a wide range of metabolic processes
which are crucial to the maintenance and production of the animal. Since the
liver and the gut are considered to function as an integrated unit, they are
often referred to as the splanchnic bed (Stangassinger and Giesecke, 1986).
A large proportion of most nutrients passing through the liver is subjected to
further metabolism.
Metabolism of acetate and butyrate
Of the three major VFAs, only acetate is not metabolised by the liver to an
appreciable extent (Leng and Annison, 1963; Cook and Miller, 1965; Berg-
man and Wolff, 1971 ). This appears in part to be due to a low activity of
acetyl-CoA synthetase in this organ under physiological conditions (Smith,
1971; Ash and Baird, 1973). The activity of acetyl-CoA hydrolase in sheep
liver, which is higher than that of acetyl-CoA synthetase, may be relevant also
in this respect (Knowles et al., 1974; Bell, 1981; Annison, 1984). On the basis
of studies with liver slices incubated with acetate as the only VFA in the me-
206 M.F.~. van Houtert /Animal Feed Science and Technology 43 (1993) 189-225
dium, Hanson and Ballard (1967) reported on the high activity of acetyl-
CoA synthetase. Subsequently, both propionate and butyrate have been shown
to depress the activating capacity of acetate in the liver by at least half (Ash
and Baird, 1973 ). Proportionately only 0.02 of all acetate passing through the
liver was metabolised in maintenance-fed sheep (Bergman and Wolff, 1971 ).
However, on a molar basis, more acetate than any other VFA is taken up by
the liver of ruminants fed roughages (Cook and Miller, 1965 ). A significant
proportion of acetate was taken up by the liver in the study by Pethick et al.
( 1981 ), using forage-fed sheep. The acetate which is taken up by the liver is
used mainly for anabolic purposes such as de novo synthesis of LCFAs, syn-
thesis of cholesterol and porphyrins (Bloch, 1947; Bell, 1981 ) and carotenes
(Van der Walt, 1984). Only minor amounts of acetate are oxidised directly
to CO2 in the liver (Pethick et al., 1981 ).
Generally, little butyrate escapes metabolism in rumen epithelium and as a
result only small amounts are absorbed into the portal blood. Butyrate reach-
ing the liver is almost completely metabolised (Annison et al., 1957; Bergman
and Wolff, 1971 ), owing to the high activity of butyryl-CoA synthetase and
other relevant enzymes in this tissue (Ash and Baird, 1973 ). The end prod-
ucts of this metabolism are acetyl-CoA, longer chain fatty acids and ketone
bodies, mainly p-hydroxybutyrate (Katz and Bergman, 1969; Bell, 1981;
Bergman, 1990).
Propionate and gluconeogenesis
In roughage-fed ruminants, a large proportion of all the propionate which

is produced in the rumen reaches the portal circulation. In the liver, propion-
ate is metabolised almost completely (Bergman and Wolff, 1971 ), with little
reaching peripheral tissues (tissues other than the splanchnic bed). Concen-
trations of propionate in portal blood of hay-fed sheep are in the range of 0.1-
0.4 mM, while that in carotid and jugular blood is only 0.01-0.02 mM (An-
nison et al., 1957; Bergman, 1975). Gluconeogenesis appears to be the main
pathway of propionate metabolism in the liver and propionate is the major
glucose precursor in roughage-fed ruminants (e.g. Cridland, 1983 ).
Glucose is essential in cellular metabolism, in particular for metabolism in
the central nervous system, blood cells, spermatozoa, the mammary gland,
the uterus and foetus, in muscle metabolism and in fat synthesis (Leng, 1970b;
Bergman, 1973 ). Glucose synthesis in the liver has been shown to account for
about 0.87 of all glucose synthesised in fed sheep, whereas the kidneys ac-
counted for 0.09 of the total glucose production (Bergman, 1973; Bergman et
al., 1974).
Gluconeogenesis from propionate commences with its activation to pro-
pionyl-CoA and its conversion via methylmalonyl-CoA into succinyl-CoA,
which enters the citric acid cycle to yield oxaloacetate. This is then translo-
M.EJ. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 207
cated from the mitochondrion into the cytoplasm in the form of malate and
then decarboxylated and phosphorylated to triose phosphate. Two molecules
of triose phosphate are then condensed to hexose phosphate which is then
converted to glucose via a number of steps (Ballard et al., 1969; Madsen,
1983c).
Estimation of the precise contribution of propionate and other glucogenic
precursors to glucose synthesis is technically difficult. Dilution of tracers
(metabolites labelled with stable or radioactive isotopes) has been the method
used most commonly to estimate the dynamics and transfer quotients of spe-
cific metabolites, both in vitro and in vivo. The interpretation of results ob-
tained by this method is complicated by a number of phenomena, in particu-
lar in studies measuring the contribution of propionate to glucose synthesis.
These include varying degrees of equilibration, recycling and interaction with
citric acid cycle intermediates (metabolic cross-over) of labelled atoms from
the tracer. The uptake of precursors for glucose synthesis by the liver can be
estimated using arterial-venous concentration differences and rates of blood
flow. This can be used to estimate the contribution of glucogenic precursors
to glucose synthesis. A combination of the two methods has also been used.
Despite the difficulties outlined above, a great number of workers have made
estimates of gluconeogenesis from various precursors in the whole body and
specific organs under a range of nutritional and physiological conditions (for
a review see Leng, 1970b; Bergman, 1973, 1983; Elliot, 1980). As much as
0.88 of all the glucose synthesised has been reported to be derived from pro-
pionate, in sheep with a low rate of gluconeogenesis as a result of infusion of
glucose (Judson and Leng, 1973). Rowe et al. (1978) estimated the parti-
tioning of propionate carbon in sheep fed a molasses-based diet using a four-
pool model. Proportionately 0.53 of all rumen propionate was oxidised to
carbon dioxide, which included intraruminal oxidation (i.e. pre-absorption)
and oxidation by the rumen epithelium and liver, while 0.46 of the propion-
ate was incorporated into blood glucose. Bergman (1983), reviewing re-
search using estimates obtained with isotope-dilution methods and also with
the arterial-venous method, stated that typically 0.27-0.40 of glucose synthe-
sis is derived from propionate. However, equilibration, recycling and meta-
bolic cross-over of labelled atoms in tracer dilution studies can seriously un-
derestimate the true contribution of various precursors to glucose synthesis
(Hetenyi, 1982; Cridland, 1983 ).
Little is known of the fate of the propionate which is not used for hepatic
gluconeogenesis. Presumably this is in part oxidised in the citric acid cycle
and in part incorporated into amino acids and other intermediates (Elliot,
1980). Very high rates of absorption of propionate from the digestive tract
may exceed the capacity of the liver to metabolise propionate (McClymont,
1951 a; Cook and Miller, 1965; Garton et al., 1972; Orskov, 1977; Jenkins and
Thonney, 1988 ). In that case, concentrations of propionate in the peripheral
circulation can increase significantly and propionate becomes available for

metabolism in other tissues (see below).
Other glucogenic precursors
Of the other glucogenic precursors, circulating amino acids, lactate and

glycerol are the three most important. Minor contributions to gluconeogene-
sis may come from other precursors such as isobutyrate, valerate and ribose
(Bergman, 1983; Giesecke, 1983 ). Acetate, butyrate and longer chain fatty
acids do not contribute to glucose synthesis, because these substrates enter
the citric acid cycle as acetyl-CoA. No net increase in oxaloacetate occurs ow-
ing to the two decarboxylation reactions in the initial stage of the citric acid
cycle (Bergman, 1983 ).
For detailed discussions of the importance of other glucogenic precursors
in gluconeogenesis in ruminants, the reader is referred to Krebs (1964),
Lindsay ( 1980 ), Bergman ( 1983 ) and Madsen (1983c), amongst others.
Metabolism of volatile fatty acids in peripheral tissues and portal-drained

viscera
Metabolism of acetate
Of the three major VFAs which are produced in the rumen, only acetate
reaches the peripheral tissues in significant quantities. Metabolism of acetate
by peripheral tissue is rapid, with a half-life of 1-3 min, hence the low con-
centrations of acetate in plasma (Annison and Lindsay, 1961; Giesecke,
1983). Bergman and Wolff ( 1971 ) estimated from studies with sheep fed 800
g day-t alfalfa pellets, that as much as 0.75 of all acetate entering the blood is
utilised by peripheral tissues. Muscle and adipose tissue were proposed to be
the two major tissues for utilisation of acetate in non-lactating ruminants.
McClymont ( 195 la,b, 1952) suggested that acetate reaching the peripheral
circulation is partitioned mainly between oxidation and fat synthesis.
Metabolism of acetate in muscle tissue

Acetate taken up by muscle cells is largely oxidised to C O 2 (Bell, 1981;
Pethick et al., 1981 ). Transport of acetate into muscle cell cytoplasm and
mitochondria is by passive diffusion, the rate of which is determined by the
concentration of acetate in blood plasma (Madsen, 1983a). For the genera-
tion ofATP, intramitochondrial acetate is then activated to acetyl-CoA under
the influence of acetyl-CoA synthetase. This process requires two high-energy
phosphate bonds per mole of acetate (Madsen, 1983b). Subsequent genera-
tion of most of the ATP commences with the oxidation of acetyl-CoA and
other intermediary metabolites in the citric acid cycle in the mitochondrial
matrix. This process generates reduced coenzymes, which in turn enter the
inner mitochondrial membrane and the electron transport chain of reactions
which is coupled to oxidative phosphorylation to generate ATP. Substrate-
level phosphorylation contributes little to the total rate of generation of ATP.
Regulation of the acetate-carbon flow into the citric acid cycle is complex.
Oxaloacetate is required for the entry of acetyl-CoA into the citric acid cycle.
The availability of oxaloacetate relative to acetyl-CoA has been proposed as
an important factor determining the rate of oxidation of acetyl-CoA (Bell,
1981 ). This has long been disputed (e.g. Lindsay, 1961 ) however, and the
balance between coenzymes and reduced coenzymes in the mitochondrial
matrix may play a more important role in this respect. For example, a high
intramitochondrial concentration of succinyl-CoA and molecules such as re-
duced nicotinamide adenine dinucleotide phosphate (NADPH) and NADH
play a role in moderating the rate of carbon flux through the citric acid cycle
(Lehninger, 1982).
The net yield of ATP from oxidative phosphorylation per mole of acetate
is 10 mol (Madsen, 1983a). Most of the ATP generated in resting muscle cells
is presumably used for protein synthesis. This requirement for ATP is low,
compared with that required during exercise.
Although acetate is the major substrate taken up by resting muscle tissue
(moles per unit time), rates of oxygen consumption for catabolism of acetate,
non-esterified LCFAs and ketone bodies are similar (Madsen, 1983a). Dur-
ing exercise, non-esterified LCFAs and glucose become the major substrates
for catabolism (Judson et al., 1976; Bird et al., 1981 ).
Metabolism of acetate in adipose tissue

Transport of acetate into adipose cells and its subsequent activation are as
described for muscle cells (Madsen, 1983b). The use of acetate as a carbon
source in the synthesis of LCFAs is the prevailing pathway of acetate metab-
olism in adipocytes. However, the rate of fatty acid synthesis in adipose tissue
of ruminants is relatively low (McClymont, 1952; Ballard et al., 1969; Lind-
say, 1983 ). Lindsay (1970) suggested that in sheep fed a maintenance ration,
synthesis of LCFAs from acetate was only 5-7 g day- ~. Sheep fed lucerne hay
and infused with ~4C-acetate intravenously for 5 days, appeared to have in-
corporated only around 0.01 of all ~4C into body lipids, regardless of their
level of feed intake (Broad and Ulyatt, 1980). Sheep injected with laC-ace-
rate into the jugular vein and slaughtered after 15 or 30 min had incorporated
about 0.08 of all injected ~4C in adipose tissues and the gut (Ingle et al.,
1972b). Similarly, Prior (1978) infused sheep intravenously with laC-ace-
tate for 180 min and was able to account for the incorporation of 0.05-0.12
of all ~4C into body fat. Presumably a large proportion of the acetate was used
for oxidation in the citric acid cycle in tissues other than adipose tissue in
these sheep.
2 10 M.F.J. van Houtert/Animal Feed Science and Technology 43 (1993) 189-225
The biochemical reactions in fatty acid synthesis in ruminants have been

reviewed in detail (Bauman, 1976; Vernon, 1981; Madsen, 1983b). In short,
adipocyte cytoplasm is the major site and acetyl-CoA is the major carbon
source for fatty acid synthesis in non-lactating ruminants (e.g. Hanson and
Ballard, 1967; Hood et al., 1972; Ingle et al., 1972a,b). As much as 0.9 of all
fatty acid synthesis in non-lactating ruminants takes place in the adipocytes
(Bauman and Davis, 1975; Annison, 1984). Lactate may, under some con-
ditions, become an important carbon source in ruminants fed grain-based diets
(Prior, 1978; Smith and Crouse, 1984; Smith and Prior, 1986). Under par-
ticular dietary conditions, propionate and butyrate (see below) and glucose
may reach the peripheral circulation in elevated amounts and then become
significant carbon sources in fatty acid synthesis (Ballard et al., 1972; Ver-
non, 1981 ). Prior to fatty acid synthesis, most of the cytoplasmic acetyl-CoA
is carboxylated to malonyl-CoA, a process which requires the hydrolysis of
ATP into adenosine diphosphate (ADP). Synthesis of fatty acids then com-
mences with the elongation of a primer molecule (generally acetyl-CoA but
at times fl-hydroxybutyryl-CoA; Bauman et al., 1970) with two-carbon units,
which are derived from the decarboxylation of malonyl-CoA. This elongation
is invoked by the fatty acid synthetase enzyme complex. Palmitate, the nor-
mal end product of this enzyme complex, can then be elongated and/or un-
saturated to yield other LCFAs. The rate-limiting step in fatty acid synthesis
in ovine adipose tissues is believed to be the carboxylation of acetyl-CoA into
malonyl-CoA (Smith and Prior, 1981 ). Fatty acid synthetase may be the rate-
limiting enzyme system in bovine adipose tissue (Smith and Prior, 1986).
The synthesis of 1 mol palmitate from 8 mol of acetate requires energy in
the form of 23 mol ATP and a source of hydrogen in the form of 14 mol of
NADPH (Vernon, 1981; Madsen, 1983b). Most of the ATP will be generated
through the oxidation of acetyl-CoA in the citric acid cycle of adipocyte mi-
tochondria. The ATP in the mitochondria is translocated into the cytoplasm
by exchange with cytoplasmic ADP through the action of ADP-ATP translo-
case in the mitochondrial membrane (Madsen, 1983a). As indicated earlier,
1 tool of acetate generates l0 mol of ATP, hence some 2.3 tool acetyl-CoA
will have to be oxidised in the citric acid cycle for every 8 mol acetate incor-
porated into palmitate.
In ruminants fed roughage-based diets, most of the required NADPH is
generated from the oxidation of glucose in the pentose phosphate cycle (Arm-
strong and Blaxter, 1957; Hanson and Ballard, 1967; Ballard et al., 1969).
Although NADPH may also be produced from the flow of acetate-carbon
through the isocitrate cycle (Bauman et al., 1970, 1973; Ingle et al., 1972a),
it appears that this pathway is of little importance in adipocytes. Generation
of NADPH from acetate via the isocitrate pathway produces ATP for which
there appears to be limited use in the adipocyte (Bauman and Davis, 1975 ).
Generation of NADPH via the pentose phosphate pathway, however, has been
M.F.J. van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 21 1
suggested to have a net requirement for ATP owing to the intensive recycling
of triose phosphate-carbon (Yang and Baldwin, 1973; Bauman and Davis,
1975 ). Given the requirements for ATP in fatty acid synthesis, however, it is
not clear whether ATP utilisation ever does become rate-limiting for this pro-
cess (Vernon, 1981 ). Preston and Leng ( 1987 ), assuming that 7 mol of ATP
are generated from 1 mol of acetate entering the isocitrate cycle, calculated
that if 0.7 of all NADPH is generated from glucose in the pentose phosphate
cycle and 0.3 from acetate in the isocitrate cycle, then the ATP generated in
the latter pathway is equivalent to that required for fatty acid synthesis. The
outcome of such calculations depends on the rate of generation of ATP from
the isocitrate pathway which has not been established unequivocally, but may
be lower. Nevertheless, present data suggest that glucose oxidation in the pen-
tose phosphate cycle provides the bulk of the reducing equivalents for fatty
acid synthesis from acetate. Glucose also furnishes virtually all the carbon for
glycerol 3-phosphate, which is required for the esterification of three LCFA
molecules into fat (Hood et al., 1972; Vernon, 1981 ).
In rat liver the N A D P H / N A D P ratio is an important regulator of carbon
flow through the pentose phosphate cycle. Whenever increased demand for
NADPH (e.g. during fat synthesis) reduces the NADPH/NADP ratio, an
increased flux of carbon through the pentose phosphate cycle is automatically
induced (Oddy and Lindsay, 1986 ). This mechanism may also be functional
in ruminants, and Thornton and Tume ( 1984, 1987 ) suggested that NADPH
synthesis may be consequential rather than causal for lipogenesis. It can be
concluded that the importance of NADPH supply in the control of the rate of
fat synthesis and acetate utilisation still appears to be unresolved.
Metabolism of acetate in the portal-drained viscera

Pethick et al. ( 1981 ) reported that in sheep fed with grass hay, the net util-
isation of acetate by the muscle, portal-drained viscera and liver accounted
for 0.27, 0.25 and 0.17, respectively, of the total net utilisation of acetate by
the sheep (see corrections published by Lindsay et al., 1986). From these
data it is clear that utilisation of acetate by the tissues of the splanchnic bed
can be considerable. Presumably, adipose tissue would utilise most of the re-
maining acetate (about 0.3 of the total net utilisation ). Net utilisation of ace-
tate by the portal-drained viscera of the sheep used by Bergman and Wolff
( 1971 ) was 0.22 of the acetate utilisation by the total body, and the authors
speculated that most of this acetate may have been used for fat synthesis.
Other uses of acetate by the portal-drained viscera may include oxidation in
smooth muscle, conversion into other compounds or use in simple exchange
reactions. In the study by Pethick et al. ( 1981 ), most of the acetate which was
taken up by the portal-drained viscera was directly oxidised to CO2, which is
in line with its high metabolic requirements (Webster, 1980; Huntington,
1990; Johnson et al., 1990).
Utilisation of propionate and butyrate
While under normal conditions little propionate escapes metabolism in the

liver in roughage-fed ruminants, the small amounts that do escape are meta-
bolised by various peripheral tissues (McClymont, 1951a). The mammary
gland of lactating ruminants is the main tissue for the use of circulating pro-
pionate. Very high levels of absorption of propionate appear to exceed the
capacity of the liver to metabolise it and may result in significant amounts
reaching peripheral tissues. Oxidation of propionate in the citric acid cycle of
adipocyte mitochondria (Hood et al., 1972; Vernon, 1981 ) and the incorpo-
ration of propionate into tissue fatty acids has been reported (Garton et al.,
1972; Duncan and Garton, 1978; Scaife et al., 1978 ), but very little propion-
ate was shown to be used as a carbon source for fatty acid synthesis in in vitro
studies with adipose tissue from cattle (Hood et al., 1972).
The amount of butyrate reaching peripheral tissues is negligible in normal
roughage-fed ruminants. The butyrate which is taken up by the peripheral
tissues and portal-drained viscera can be oxidised to carbon dioxide or can be
used as a carbon source for lipogenesis (Annison et al., 1963 ).
Energetic efficiency of utilisation of roughage based and concentrate-based

diets
Roughage-based diets have been observed to be used with a lower energetic

efficiency than concentrate-based diets for growth in ruminants (see Blaxter
and Boyne, 1978). An early hypothesis put forward to explain this was that
the energy needed for digestive processes was higher for roughage-based diets
than for concentrate-based diets. The idea appears to have been derived from
work by Von Mering and Zuntz (1877) who found that metabolism of ab-
sorbed nutrients alone could not explain the magnitude of increases in heat
production in fed rabbits as compared with starved rabbits. These workers
suggested that the increased heat production could be caused by an increase
in the physical and glandular activity of the gastrointestinal tract associated
with absorption and digestion.
With the discovery of the key role of VFAs in oxidative and synthetic me-
tabolism in ruminants, an alternative hypothesis involving metabolism of
VFAs has evolved. The proportion of acetate in the VFAs which are produced
in the rumen has been found to be negatively correlated with the efficiency
with which the energy is retained by the ruminant (Elliot and Loosli, 1959;
and Allen, 1966c; Blaxter, 1967 ). Conversely, a positive correlation has been
reported between the proportion of propionate in the VFAs and the efficiency
of energy retention. However, mixtures of the three main VFAs with molar
proportions of acetic acid varying from 0.25 to 0.9 were retained with similar
high energetic efficiencies (0.85-0.90) when infused intraruminally over a
period of 1 week in fasting sheep (Armstrong et al., 1957). Similar trials with
fattening sheep showed that a mixture of acetic, propionic and butyric acids
of 75:15:10 was used with only half the energetic efficiency of a mixture of
25:45:30 (Armstrong et al., 1958).
As discussed earlier, lipogenesis from acetate requires oxidation of glucose
in the pentose phosphate pathway to generate most of the required reduced
coenzyme (NADPH). Glucose also provides glycerol for esterification of
LCFAs. This interdependence of the availability of acetate and glucose has
been used to formulate the theory that a high intracellular ratio of acetate to
glucose in adipocytes would not provide sufficient NADPH to synthesise all
acetate into LCFAs. This would thereby limit a potentially important sink for
acetate, which would either limit potential feed intake or could result in en-
ergetically wasteful use of acetate in heat-generating substrate cycles (Arm-
strong and Blaxter, 1957; Armstrong, 1965; Bauman and Davis, 1975; Agri-
cultural Research Council, 1980; MacRae and Lobley, 1982 ).
Propionate is the main glucogenic precursor in ruminants. The above hy-
pothesis therefore implies a minimum requirement of propionate relative to
acetate in nutrients which are available above the requirements for tissue
maintenance and protein deposition, for efficient utilisation of acetate in syn-
thesis of LCFAs. This requirement has been calculated, from theoretical con-
siderations, as 1 mol propionate per 4.1 mol acetate, if all reduced coenzyme
is generated in the pentose phosphate cycle (see Orskov et al., 1979). The
influence of different ratios of acetate to propionate on the energetic effi-
ciency of growth, has been the subject of a large number of studies. Calori-
metric or serial-slaughtering techniques have been used to estimate the effi-
ciency with which VFAs, either infused into the rumen or added to the basal
diet, were retained as tissue energy. There is now an abundance of apparently
conflicting published research on this issue. Published results demonstrate
that under physiological conditions, acetate may be used with a lower ener-
getic efficiency when propionate is in short supply (Poole and Allen, 1970;
Hovell et al., 1976; Hovell and Greenhalgh, 1978; Tyrrell et al., 1979; Lobley
and MacRae, 1987; Jenkins and Thonney, 1988; Abdul-Razzaq et al., 1988 ).
Others reported that the efficiency of energy utilisation from VFAs did not
vary with the ruminal ratios of acetate and propionate, over a range of VFA
ratios occurring under practical conditions (Essig et al., 1958; Rook et al.,
1963; Elliot et al., 1965; ¢0rskov and Allen, 1966a,b,c; Orskov et al., 1966; Bull
et al., 1970; Lawrence and Thomas, 1973; Orskov et al., 1979).
Several workers (Hovell et al., 1976; Reid et al., 1980; MacRae and Lobley,
1982, 1986) have suggested that the provision of high levels of dietary glu-
cogenic precursors other than propionate (e.g. non-essential amino acids and/
or glycerol) may account for most of those reports where the utilisation of
acetate did not depend upon the presence of propionate within physiological
ranges. Sheep given diets with a high protein content, or sheep infused with
casein per abomasum, may have absorbed enough protein to allow sufficient
gluconeogenesis from amino acids to ensure efficient utilisation of acetate.
Egan ( 1965 ) reported that the infusion of casein into the duodenum of sheep
offered a roughage-based diet increased their ability to cope with an acetate
load (5 mmol kg- 1liveweight). Feeding acetate as triacetin (Bull et al., 1970;
Johnson et al., 1973 ) provides the animal with glucogenic glycerol, which is
sufficient to ensure efficient utilisation of acetate. MacRae and Lobley (1986)
have suggested that the efficiency of utilisation of energy from roughage-based
diets for growth may be influenced by the availability of amino acids (to pro-
vide NADPH) if the ratio of absorbed acetate to propionate is greater than
3.5. In some of the experiments where acetate was used less efficiently than
propionate, however, a basal diet with a high protein content was offered in
the form of dried grass or a grain-based diet supplemented with additional
protein meal.
After more than a century of research, the reasons for the generally lower
efficiency of utilisation of dietary energy derived from roughage-based diets,
as compared with concentrate-based diets, are still equivocal and an issue of
debate. There are indications that the increased time spent eating and rumi-
nating with roughage-based diets is of minor consequence in the energy me-
tabolism of the ruminant (Webster et al., 1976; Webster, 1980). Neverthe-
less, time spent eating and ruminating and the lower rates of degradation and
outflow from the rumen have been re-emphasised 03rskov and MacLeod,
1990) as the main causes of the low efficiency of utilisation of dietary energy
from roughage-based diets compared with grain-based diets. Increases in the
nutrient requirements of the gastrointestinal tract due to increases in its rela-
tive mass in roughage-fed ruminants may play an important role also (see
Southon et al., 1985; Burrin et al., 1990). Recent work by Reynolds et al.
( 1991 ) suggests that all of these factors may, to a large extent, explain the
lower efficiency of utilisation of dietary energy derived from roughage-based
diets, compared with concentrate-based diets. Important factors in that study
were increased metabolic activity per unit of tissue of the gastrointestinal tract
and increased total tissue weight, as well as physical work of digestion and
absorption.
Acknowledgements
I would like to thank R.A. Leng and J.V. Nolan for valuable suggestions.
References
Abdul-Razzaq, H.A., Bickerstaffe, R. and Savage, G.P., 1988. The influence of rumen volatile
fatty acids on blood metabolites and body composition of growing lambs. Aust. J. Agric.
Res., 39:505-515.
M.F..L van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 215
Agricultural Research Council, 1980. The Nutrient Requirements of Ruminant Livestock.

Commonwealth Agricultural Bureaux, Farnham Royal, 351 pp.
Allan, S.A. and Miller, E.L., 1972. The effect of urea supplementation on the nitrogen reaching
the abomasum of lambs and on the extent of nitrogen recycling. Proc. Nutr. Soc., 31: 26a.
(Abstr.)
Annison, E.F., 1954. Some observations on volatile fatty acids in the sheep's rumen. Biochem.
J., 57: 400-405.
Annison, E.F., 1984. The metabolism of neutral and acidic lipids by tissues of the ruminant. In:
F.M.C. Gilchrist and R.I. Mackie (Editors), Herbivore Nutrition in the Subtropics and
Tropics. The Science Press, Craighall, South Africa, pp. 549-570.
Annison, E.F. and Armstrong, D.G., 1970. Volatile fatty acid metabolism and energy supply.
In: A.T. Phillipson (Editor), Physiology of Digestion and Metabolism in the Ruminant. Or-
iel Press, Newcastle-upon-Tyne, UK, pp. 422-437.
Annison, E.F. and Lewis, D., 1959. Metabolism in the Rumen. Methuen and Co., London, 185
PP.
Annison, E.F. and Lindsay, D.B., 1961. Acetate utilization in sheep. Biochem. J., 78: 777-785.
Annison, E.F., Hill, K.J. and Lewis, D., 1957. Studies on the portal blood of sheep. 2. Absorp-
tion of volatile fatty acids from the rumen of the sheep. Biochem. J., 66: 592-599.
Annison, E.F., Leng, R.A., Lindsay, D.B. and White, R.R., 1963. The metabolism of acetic acid,
propionic acid and butyric acid in sheep. Biochem. J., 88: 248-252.
Annison, E.F., Brown, R.E., Leng, R.A., Lindsay, D.B. and West, C.E., 1967. Rates of entry and
oxidation of acetate, glucose, D (-)-fl-hydroxybutyrate, palmitate, oleate and stearate, and
rates of production and oxidation of propionate and butyrate in fed and starved sheep.
Biochem. J., 104: 135-147.
Argyle, J.L. and Baldwin, R.L., 1989. Effects of amino acids and peptides on rumen microbial
growth yields. J. Dairy Sci., 72:2017-2027.
Armstrong, D.G., 1965. Carbohydrate metabolism in ruminants and energy supply. In: R.W.
Dougherty (Editor), Physiology of Digestion in the Ruminant. Butterworths, Washington
DC, pp. 272-288.
Armstrong, D.G. and Blaxter, K.L., 1957. The heat increment of steam-volatile fatty acids in
fasting sheep. Br. J. Nutr., 11: 247-272.
Armstrong, D.G., Blaxter, K.L. and Graham, N.McC., 1957. The heat increments of mixtures
of steam-volatile fatty acids in fasting sheep. Br. J. Nutr., 11: 392-408.
Armstrong, D.G., Blaxter, K.L., Graham, N.McC. and Wainman, F.W., 1958. The utilization
of the energy of two mixtures of steam volatile fatty acids by fattening sheep. Br. J. Nutr.,
12: 177-188.
Ash, R. and Baird, G.D., 1973. Activation of volatile fatty acids in bovine liver and rumen
epithelium. Biochem. J., 136:311-319.
Baird, G.D., Hibbitt, K.G., Hunter, G.D., Lund, P., Stubbs, M. and Krebs, H.A., 1968. Bio-
chemical aspects of bovine ketosis. Biochem. J., 107: 683-689.
Baker, F., 1942. Microbial factors in the digestive assimilation of starch and cellulose in herbi-
vora. Nature, 150: 479-480.
Baldwin, R.L., 1965. Pathways of carbohydrate metabolism in the rumen. In: R.W. Dougherty
(Editor), Physiology of Digestion in the Ruminant. Butterworths, Washington DC, pp. 379-
389.
Ballard, F.J., Hanson, R.W. and Kronfeld, D.S., 1969. Gluconeogenesis and lipogenesis in tis-
sue from ruminant and non-ruminant animals. Fed. Proc., 28:218-231.
Ballard, F.J., Filsell, O.H. and Jarrett, I.G., 1972. Effects of carbohydrate availability on lipo-
genesis in sheep. Biochem. J., 126: 193-200.
Barcroft, J., McAnally, R.A. and Phillipson, A.T., 1944. Absorption of volatile fatty acids from
the alimentary tract of the sheep and other animals. J. Exp. Biol., 20:120-129.
216 M.~:Z van Houtert /Animal FeedScienceand Technology43 (1993.) 189-225
Bauchop, T. and Elsden, S.R., 1960. The growth of micro-organisms in relation to their energy
supply. J. Gen. Microbiol., 23: 457-469.
Bauman, D.E., 1976. Intermediary metabolism of adipose tissue. Fed. Proc., 35:2308-2313.
Bauman, D.E. and Davis, C.L., 1975. Regulation of lipid metabolism. In: I.W. McDonald and
A.C.I. Warner (Editors), Digestion and Metabolism in the Ruminant. The University of
New England, Armidale, N.S.W., pp. 496-509.
Bauman, D.E., Brown, R.E. and Davis, C.L., 1970. Pathways of fatty acid synthesis and reduc-
ing equivalent generation in mammary gland of rat, sow and cow. Arch. Biochem. Biophys.,
140: 237-244.
Bauman, D.E., Mellenberger, R.W. and Derrig, R.G., 1973. Fatty acid synthesis in sheep mam-
mary tissue. J. Dairy Sci., 56: 1312-1318.
Beck, U., Emmanuel, B. and Giesecke, D., 1984. The ketogenic effect of glucose in rumen epi-
thelium of ovine (Ovis aries) and bovine (Bos taurus) origin. Comp. Biochem. Physiol.,
77B: 517-521.
Becker, E.R. and Everett, R.C., 1930. Comparative growths of normal and infusoria-free lambs.
Am. J. Hyg., 11: 362-370.
Bell, A.W., 1981. Lipid metabolism in liver and selected tissues and in the whole body of rumi-
nant animals. In: W.W. Christie (Editor), Lipid Metabolism in Ruminant Animals. Perga-
mon Press, Oxford, pp. 363-410.
Bergen, W.G., 1972. Rumen osmolality as a factor in feed intake control of sheep. J. Anita. Sci.,
34: 1054-1060.
Bergman, E.N., 1973. Glucose metabolism in ruminants as related to hypoglycemia and ketosis.
Cornell Vet., 63: 341-382.
Bergman, E.N., 1975. Production and utilization of metabolites by the alimentary tract as mea-
sured in portal and hepatic blood. In: I.W. McDonald and A.C.1. Warner (Editors), Diges-
tion and Metabolism in the Ruminant. The University of New England, Armidale, N.S.W.,
pp. 292-305.
Bergman, E.N., 1983. The pools of cellular nutrients: glucose. In: P.M. Riis (Editor), Dynamic
Biochemistry of Animal Production. Elsevier, Amsterdam, pp. 173-196.
Bergman, E.N., 1990. Energy contributions of volatile fatty acids from the gastrointestinal tract
in various species. Physiol. Rev., 70: 567-590.
Bergman, E.N. and Wolff, J.E., 1971. Metabolism of volatile fatty acids by liver and portal-
drained viscera in sheep. Am. J. Physiol., 221: 586-592.
Bergman, E.N., Reid, R.S., Murray, M.G., Brockway, J.M. and Whitelaw, F.G., 1965. Intercon-
versions and production of volatile fatty acids in the sheep rumen. Biochem. J., 97: 53-58.
Bergman, E.N., Brockman, R.P. and Kaufman, C.F., 1974. Glucose metabolism in ruminants:
comparison of whole-body turnover with production by gut, liver and kidneys. Fed. Proc.,
33: 1849-1854.
Bird, A.R., Chandler, K.D. and Bell, A.W., 1981. Effects of exercise and plane of nutrition on
nutrient utilisation by the hind limb of the sheep. Aust. J. Biol. Sci., 34: 541-550.
Bird, S.H., 1985. Rumen protozoa--potential for manipulation. In: R.B. Cumming (Editor),
Recent Advances in Animal Nutrition in Australia. Paper 34, The University of New Eng-
land, Armidale, N.S.W.
Bird, S.H., 1991. The influence of the presence of protozoa on ruminant production: A review.
In: D.J. Farrell (Editor), Recent Advances in Animal Nutrition in Australia. The University
of New England, Armidale, N.S.W., pp. 15-27.
Blaxter, K.L., 1967. The Energy Metabolism of Ruminants, 2nd edn. Hutchinson Scientific and
Technical, London, 332 pp.
Blaxter, K.L. and Boyne, A.W., 1978. The estimation of the nutritive value of feeds as energy
sources for ruminants and the derivation of feeding systems. J. Agric. Sci., 90: 47-68.
Bloch, K., 1947. The metabolism of acetic acid in animal tissues. Physiol. Rev., 27: 574-620.
Bonhomme, A., 1990. Rumen ciliates: their metabolism and relationships with bacteria and
their hosts. Anim. Feed Sci. Technol., 30:203-266.
Boniface, A.N., Murray, R.M. and Hogan, J.P., 1986. Optimum level of ammonia in the rumen
liquor of cattle fed tropical pasture hay. Proc. Aust. Soc. Anim. Prod., 16:151-154.
Broad, T.E. and Ulyatt, M.J., 1980. The effect of level of food intake on the incorporation of
acetate into lipids and its distribution among various tissues in sheep. Br. J. Nutr., 44:71-
79.
Bull, L.S., Reid, J.T. and Johnson, D.E., 1970. Energetics of sheep concerned with the utiliza-
tion of acetic acid. J. Nutr., 100: 262-276.
Bull, L.S., Rumpler, W.V., Sweeney, T.F. and Zinn, R.A., 1979. Influence of ruminal turnover
on site and extent of digestion. Fed. Proc., 38:2713-2719.
Burrin, D.G., Ferrell, C.L., Britton, R.A. and Bauer, M., 1990. Level of nutrition and visceral
organ size and metabolic activity in sheep. Br. J. Nutr., 64: 439-448.
Bush, R.S. and Milligan, L.P., 1971. Enzymes of ketogenesis in bovine rumen epithelium. Can.
J. Anim. Sci., 51: 129-133.
Coleman, G.S., 1979. The role ofrumen protozoa in the metabolism of ruminants given tropical
feeds. Trop. Anim. Prod., 4: 199-213.
Coleman, G.S., 1988. The importance of rumen ciliate protozoa in the growth and metabolism
of the host ruminant. Int. J. Anim. Sci., 3: 75-95.
Cook, R.M. and Miller, L.D., 1965. Utilization of volatile fatty acids in ruminants. I. Removal
of them from portal blood by the liver. J. Dairy Sci., 48:1339-1345.
Cridland, S.W., 1983. Studies on the flows of propionate carbon to glucose in sheep. Ph.D.
Thesis, The University of New England, Armidale, N.S.W., 171 pp.
Czerkawski, J.W., 1986. An Introduction to Rumen Studies. Pergamon Press, Oxford, 236 pp.
Czerkawski, J.W., Blaxter, K.L. and Wainman, F.W., 1966. The metabolism of oleic, linoleic
and linolenic acids by sheep with reference to their effects on methane production. Br. J.
Nutr., 20: 349-362.
Czerkawski, J.W., Christie, W.W., Breckenridge, G. and Hunter, M.L., 1975. Changes in the
rumen metabolism of sheep given increasing amounts of linseed oil in their diet. Br. J. N utr.,
24: 25-44.
DanieUi, J.F., Hitchcock, M.W.S., Marshall, R.A. and Phillipson, A.T., 1946. The mechanism
of absorption from the rumen as exemplified by the behaviour of acetic, propionic and bu-
tyric acids. J. Exp. Biol., 22: 75-84.
Demeyer, D.I., 1981. Rumen microbes and digestion of plant cell walls. Agric. Environ., 6: 295-
337.
Demeyer, D.I. and Henderickx, H.K., 1967. The effect of C~8 unsaturated fatty acids on meth-
ane production in vitro by mixed rumen bacteria. Biochim. Biophys. Acta, 137: 484-497.
Dobson, M.J., Brown, W.C.B., Dobson, A. and Phillipson, A.T., 1956. A histological study of
the organization of the rumen epithelium of sheep. Q. J. Exp. Physiol., 41: 247-252.
Duncan, W.R.H. and Carton, G.A., 1978. Differences in the proportions of branched-chain
fatty acids in subcutaneous triacylglycerols of barley-fed ruminants. Br. J. Nutr., 40: 29-33.
Durand, M. and Kawashima, R., 1980. Influence of minerals in rumen microbial digestion. In:
Y. Ruckebusch and P. Thivend (Editors), Digestive Physiology and Metabolism in Rumi-
nants. MTP Press, Lancaster, UK, pp. 375-408.
Egan, A.R., 1965. Nutritional status and intake regulation in sheep. IV. The influence of protein
supplements upon acetate and propionate tolerance of sheep fed on low quality chaffed oa-
ten hay. Aust. J. Agric. Res., 16: 473-483.
Elliot, J.M., 1980. Propionate metabolism and vitamin B12. In: Y. Ruckebusch and P. Thivend
(Editors), Digestive Physiology and Metabolism in Ruminants. MTP Press, Lancaster, UK,
pp. 485-503.
218 M.F.J. van Houtert /Animal FeedScienceand Technology43 (1993) 189-225
Elliot, J.M. and Loosli, J.K., 1959. Relationship of milk production efficiency to the relative
proportions of the rumen volatile fatty acids. J. Dairy Sci., 42: 843-848.
Elliot, J.M., Hogue, D.E., Myers, G.S. and Loosli, J.K., 1965. Effect of acetate and propionate
on the utilization of energy by growing-fattening lambs. J. Nutr., 87: 233-238.
Elsden, S.R., 1946. The application of the silica gel partition chromatogram to the estimation
of volatile fatty acids. Biochem. J., 40: 252-256.
Elsden, S.R. and Phillipson, A.T., 1948. Ruminant digestion. Annu. Rev. Biochem., 17: 705-
726.
Emmanuel, B., Stangassinger, M. and Giesecke, D., 1982. Production of D ( - )-3-hydroxybu-
tyrate by an alternate mechanism in rumen epithelium of ovine (Ovis aries) and bovine
(Bos taurus). Comp. Biochem. Physiol., 72B: 415-419.
Erfle, J.D., Sauer, F.D. and Mahadevan, S., 1986. Energy metabolism in rumen microbes. In:
L.P. Milligan, W.L. Grovum and A. Dobson (Editors), Control of Digestion and Metabo-
lism in Ruminants. Prentice-Hall, Englewood Cliffs, N J, pp. 81-99.
Essig, H.W., Hatfield, E.E. and Connor, B., 1958. Levels and ratios of volatile fatty acids as
roughage substitutes for growing-fattening lambs. J. Anita. Sci., 17:1198-1199.
Garton, G.A., Hovell, F.D.DeB. and Duncan, W.R.H., 1972. Influence of dietary volatile fatty
acids on the fatty-acid composition of lamb triglycerides, with special reference to the effect
of propionate on the presence of branched-chain components. Br. J. Nutr., 28:409-416.
Giesecke, D., 1983. The pools of cellular nutrients: plasma free fatty acids. In: P.M. Riis (Edi-
tor), Dynamic Biochemistry of Animal Production. Elsevier, Amsterdam, pp. 197-214.
Giesecke, D., Beck, U. and Emmanuel, B., 1985. Ketogenic regulation by certain metabolites in
rumen epithelium. Comp. Biochem. Physiol., 81 B: 863-867.
Hanson, R.W. and Ballard, F.J., 1967. The relative significance of acetate and glucose as pre-
cursors for lipid synthesis in liver and adipose tissue from ruminants. Biochem. J., 105: 529-
536.
Harfoot, C.G., 1978. Anatomy, physiology and microbiology of the ruminant digestive tract.
Prog. Lipid Res., 17: 1-19.
Harrison, D.G. and McAllan, A.B., 1980. Factors affecting microbial growth yields in the reti-
culo-rumen. In: Y. Ruckebusch and P. Thivend (Editors), Digestive Physiology and Metab-
olism in Ruminants. MTP Press, Lancaster, UK, pp. 205-226.
Harrison, D.G., Beever, D.E., Thomson, D.J. and Osbourn, D.F., 1975. Manipulation ofrumen
fermentation in sheep by increasing the rate of flow of water from the rumen. J. Agric. Sci.,
85: 93-101.
Hastings, E.G., 1944. The significance of the bacteria and the protozoa of the rumen of the
bovine. Bacteriol. Rev., 8: 235-254.
Henderson, C., 1973. The effects of fatty acids on pure cultures of rumen bacteria. J. Agric. Sci.,
81: 107-112.
Hespell, R.B., 1979. Efficiency of growth by ruminal bacteria. Fed. Proc., 38:2707-2712.
Hespell, R.B., 1984. Influence of ammonia assimilation pathways and survival strategy on rum-
inal microbial growth. In: F.M.C. Gilchrist and R.I. Mackie (Editors), Herbivore Nutrition
in the Subtropics and Tropics. The Science Press, Craighall, South Africa, pp. 346-358.
Hespell, R.B. and Bryant, M.P., 1979. Efficiency of rumen microbial growth: influence of some
theoretical and experimental factors of YATP. J. Anim. Sci., 49: 1640-1659.
Hetenyi, G., 1982. Correction for the metabolic exchange of '4C for 12C atoms in the pathway
of gluconeogenesis in vivo. Fed. Proc., 41: 104-109.
Hird, F.J.R. and Weidemann, M.J., 1964. Ketone-body synthesis in relation to age of lambs.
Biochem. J., 93: 423-430.
Hood, R.L., Thompson, E.H. and Allen, C.E., 1972. The role of acetate, propionate, and glucose
as substrates for lipogenesis in bovine tissues. Int. J. Biochem., 3: 598-606.
Hovell, F.D.DeB. and Greenhalgh, J.F.D., 1978. The utilization of diets containing acetate,
propionate or butyrate salts by growing lambs. Br. J. Nutr., 40:171-183.
Hovell, F.D.DeB., Greenhalgh, J.F.D. and Wainman, F.W., 1976. The utilization of diets con-
taining acetate salts by growing lambs as measured by comparative slaughter and respiration
calorimetry, together with rumen fermentation. Br. J. Nutr., 35: 343-363.
Hungate, R.E., 1966. The Rumen and its Microbes. Academic Press, London, 533 pp.
Hungate, R.E., 1975. The rumen microbial ecosystem. Annu. Rev. Ecol. Syst., 6: 39-66.
Huntington, G.B., 1990. Energy metabolism in the digestive tract and liver of cattle: influence
of physiological state and nutrition. Reprod. Nutr. Dev., 30: 35-47.
Ingle, D.L., Bauman, D.E. and Garrigus, U.S., 1972a. Lipogenesis in the ruminant: in vitro
study of tissue sites, carbon source and reducing equivalent generation for fatty acid synthe-
sis. J. Nutr., 102: 609-616.
Ingle, D.L., Bauman, D.E. and Garrigus, U.S., 1972b. Lipogenesis in the ruminant: in vivo site
of fatty acid synthesis in sheep. J. Nutr., 102:617-624.
Jenkins, T.C. and Thonney, M.L., 1988. Effect of propionate level in a volatile fatty acid salt
mixture fed to lambs on weight gain, body composition and plasma metabolites. J. Anim.
Sci., 66: 1028-1035.
Johnson, D.E., Reid, J.T., Bull, L.S. and Robb, J., 1973. Observations on the efficient utiliza-
tion of acetic acid above maintenance by sheep. In: K.H. Menke, H.J. Lantzsch and J.R.
Reichl (Editors), Energy Metabolism of Farm Animals. Dokumentationsstelle, Universit~it
Hohenheim, Germany, pp. 43-46.
Johnson, D.E., Johnson, H.A. and Baldwin, R.L., 1990. Changes in liver and gastrointestinal
tract energy demands in response to physiological workload in ruminants. J. Nutr., 120: 649-
655.
Jouany, J.P., Demeyer, D.I. and Grain, J., 1988. Effect of defaunating the rumen. Anita. Feed
Sci. Technol., 21: 229-265.
Judson, G.J. and Leng, R.A., 1973. Studies on the control of gluconeogenesis in sheep: effect of
propionate, casein and butyrate infusions. Br. J. Nutr., 29:175-195.
Judson, G.J., Fissell, O.H. and Garrett, I.G., 1976. Glucose and acetate metabolism in sheep at
rest and during exercise. Aust. J. Biol. Sci., 29:215-222.
Kandylis, K., 1984. The role of sulphur in ruminant nutrition. A review. Livest. Prod. Sci., 1 l:
611-624.
Katz, M.L. and Bergman, E.N., 1969. Hepatic and portal metabolism of glucose, free fatty acids,
and ketone bodies in the sheep. Am. J. Physiol., 216: 953-960.
Knowles, S.E., Jarrett, I.G., Filsell, O.H. and Ballard, F.J., 1974. Production and utilisation of
acetate in mammals. Biochem. J., 142:401-411.
Koundakjian, P.P. and Snoswell, A.M., 1976. Ketone body and fatty acid metabolism in sheep
tissues. 3-Hydroxybutyrate dehydrogenase, a cytoplasmic enzyme in sheep liver and kidney.
Biochem. J., 119: 49-57.
Krebs, H.A., 1964. The metabolic fate of amino acids. In: H.N. Munro and J.B. Allison (Edi-
tors), Mammalian Protein Metabolism. Vol. 1. Academic Press, New York, pp. 125-176.
Lawrence, P.R. and Thomas, P.C., 1973. A note on the utilization of the energy of the short-
chain fatty acids in the fattening sheep. Anim. Prod., 17:209-212.
Lehninger, A.L., 1982. Principles of Biochemistry. Worth Publishers, New York, 1011 pp.
Leng, R.A., 1965. Ketone body metabolism in normal and underfed pregnant sheep and in preg-
nancy toxaemia. Res. Vet. Sci., 6: 433-441.
Leng, R.A., 1970a. Formation and production of volatile fatty acids in the rumen. In: A.T.
Phillipson (Editor), Physiology of Digestion and Metabolism in the Ruminant. Oriel Press,
Newcastle-upon-Tyne, UK, pp. 489-503.
Leng, R.A., 1970b. Glucose synthesis in ruminants. Adv. Vet. Sci., 14: 209-260.
Leng, R.A., 1973. Salient features of the digestion of pastures by ruminants and other herbi-
vores. In: G.W. Butler and R.W. Bailey (Editors), Chemistry and Biochemistry of Herbage.
Voi. 3. Academic Press, London, pp. 81-129.
Leng, R.A., 1982a. Modification of rumen fermentation. In: J.B. Hacker (Editor), Nutritional
Limits to Animal Production from Pastures. Commonwealth Agricultural Bureaux, Farn-
ham Royal, pp. 427-453.
Leng, R.A., 1982b. Dynamics of protozoa in the rumen of sheep. Br. J. Nutr., 48:399-415.
Leng, R.A., 1990. Factors affecting the utilization of'poor-quality' forages by ruminants partic-
ularly under tropical conditions. Nutr. Res. Rev., 3: 277-303.
Leng, R.A. and Annison, E.F., 1963. Metabolism of acetate, propionate and butyrate by sheep-
liver slices. Biochem. J., 86:319-327.
Leng, R.A. and Brett, D.J., 1966. Simultaneous measurements of the rates of production of
acetic, propionic and butyric acids in the rumen of sheep on different diets and the correla-
tion between production rates and concentrations of these acids in the rumen. Br. J. Nutr.,
20: 541-552.
Leng, R.A. and Leonard, G.J., 1965. Measurement of the rates of production of acetic, pro-
pionic and butyric acids in the rumen of sheep. Br. J. Nutr., 19: 469-484.
Leng, R.A. and Nolan, J.V., 1984. Nitrogen metabolism in the rumen. J. Dairy Sci., 67: 1072-
1089.
Leng, R.A. and West, C.E., 1969. Contribution of acetate, butyrate, palmitate, stearate and oleate
to ketone body synthesis in sheep. Res. Vet. Sci., 10: 57-63.
Leng, R.A., Steel, J.W. and Luick, J.R., 1967. Contribution of propionate to glucose synthesis
in sheep. Biochem. J., 103: 785-790.
Lindsay, D.B., 1961. Endocrine control of metabolism in ruminants. In: D. Lewis (Editor),
Digestive Physiology and Nutrition of the Ruminant. Butterworths, London, pp. 235-241.
Lindsay, D.B., 1970. Carbohydrate metabolism in ruminants. In: A.T. Phillipson (Editor),
Physiology of Digestion and Metabolism in the Ruminant. Oriel Press, Newcastle-upon-Tyne,
UK, pp. 438-451.
Lindsay, D.B., 1980. Amino acids as energy sources. Proc. Nutr. Soc., 39: 53-59.
Lindsay, D.B., 1983. Growth and fattening. In: J.A.F. Rook and P.C. Thomas (Editors), Nutri-
tional Physiology of Farm Animals. Longman, London, pp. 261-313.
Lindsay, D.B., Pethick, D.W., Barker, P.J. and Northrop, A.J., 1986. Errors in blood flow mea-
surements (Letters to the Editors). Br. J. Nutr., 56: 313.
Lobley, G.E. and MacRae, J.C., 1987. Acetate utilization in sheep. In: P.W. Moe, H.F. Tyrrell
and P.J. Reynolds (Editors), Energy Metabolism of Farm Animals. Rowman and Littlefield,
Totowa, N J, pp. 38-41.
Mackie, R.I. and Therion, J.J., 1984. Influence of mineral interactions on growth efficiency of
rumen bacteria. In: F.M.C. Gilchrist and R.I. Mackie (Editors), Herbivore Nutrition in the
Subtropics and Tropics. The Science Press, Craighall, South Africa, pp. 455-477.
Mackie, R.I. and White, B.A., 1990. Recent advances in rumen microbial ecology and metabo-
lism: potential impact on nutrient output. J. Dairy Sci., 73: 2971-2995.
MacRae, J.C. and Lobley, G.E., 1982. Some factors which influence thermal energy losses dur-
ing the metabolism of ruminants. Livest. Prod. Sci., 9: 447-456.
MacRae, J.C. and Lobley, G.E., 1986. Interactions between energy and protein. In: L.P. Milli-
gan, W.L. Grovum and A. Dobson (Editors), Control of Digestion and Metabolism in Ru-
minants. Prentice-Hall, Englewood Cliffs, NJ, pp. 367-385.
Madsen, A., 1983a. The molecular basis of animal production: metabolism in skeletal muscle
cells. In: P.M. Riis (Editor), Dynamic Biochemistry of Animal Production. Elsevier, Am-
sterdam, pp. 9-28.
Madsen, A., 1983b. The molecular basis of animal production: metabolism in adipose cells. In:
P.M. Riis (Editor), Dynamic Biochemistry of Animal Production. Elsevier, Amsterdam, pp.
29-52.
M.F.£ van Houtert /Animal Feed Science and Technology 43 (1993) 189-225 221
Madsen, A., 1983c. The molecular basis of animal production: metabolism in liver cells. In:
P.M. Riis (Editor), Dynamic Biochemistry of Animal Production. Elsevier, Amsterdam, pp.
53-74.
Maeng, W.J., van Nevel, C.J., Baldwin, R.L. and Morris, J.G., 1976. Rumen microbial growth
rates and yields: effect of amino acids and protein. J. Dairy Sci., 59: 68-79.
Maeng, W.J., Chang, M.B. and Yun, H.S., 1989. Dilution rates on the efficiency of rumen mi-
crobial growth in continuous culture. Asian-Australas. J. Anim. Sci., 2: 477-480.
Mall6vre, A., 1891. Der Einfluss der als G~ihrungsprodukt der Cellulose gebildeten essigs~iure
auf den Gaswechsel. Pfluegers Archiv., 49: 460-477.
McAnally, R.A., 1944. The determination of total volatile fatty acids in blood. J. Exp. Biol., 20:
130-131.
McClymont, G.L., 195 l a. Identification of the volatile fatty acid in the peripheral blood and
rumen of cattle and the blood of other species. Aust. J. Agric. Res., 2: 92-103.
McClymont, G.L., 195 lb. Volatile fatty acid metabolism of ruminants, with particular refer-
ence to the lactating bovine mammary gland and the composition of milk fat. Aust. J. Agric.
Res., 2: 158-180.
McClymont, G.L., 1952. Specific dynamic action of acetic acid and heat increment of feeding
in ruminants. Aust. J. Sci. Res. Ser. B, 5: 374-383.
McGarry, J.D. and Foster, D.W., 1980. Regulation of hepatic fatty acid oxidation and ketone
body production. Annu. Rev. Biochem., 49: 395-420.
Mehrez, A.Z., Orskov, E.R. and McDonald, I., 1977. Rates of rumen fermentation in relation
to ammonia concentration. Br. J. Nutr., 38: 437-443.
Miller, E.L., 1973. Evaluation of foods as sources of nitrogen and amino acids. Proc. Nutr. Soc.,
32: 79-84.
Miller, T.L. and Jenesel, S.E., 1979. Enzymology of butyrate formation by Butyrivibriofibrisol-
vens. J. Bacteriol., 138: 99-104.
Mould, F.L. and Orskov, E.R., 1983. Manipulation of rumen fluid pH and its influence on
cellulolysis, in sacco dry matter degradation and the rumen microflora of sheep offered either
hay or concentrate. Anim. Feed Sci. Technol., 10: 1-14.
Mould, F.L., Orskov, E.R. and Mann, S.O., 1983. Associative effects of mixed feeds. I. Effects
of type and level of supplementation and the influence of rumen fluid pH on cellulolysis in
vivo and dry matter digestion of various roughages. Anim. Feed Sci. Technol., 10:15-30.
Nolan, J.V. and Leng, R.A., 1972. Dynamic aspects of ammonia and urea metabolism in sheep.
Br. J. Nutr., 27: 177-194.
Oddy, V.H. and Lindsay, D.B., 1986. Metabolic and hormonal interactions and their potential
effects on growth. In: P.J. Buttery, D.B. Lindsay and N.B. Haynes (Editors), Control and
Manipulation of Animal Growth. Butterworths, London, pp. 231-248.
Okorie, A.U., Buttery, P.J. and Lewis, D., 1977. Ammonia concentration and protein synthesis
in the rumen. Proc. Nutr. Soc., 36: 38a. (Abstr.)
Orskov, E.R., 1975. Manipulation ofrumen fermentation for maximum food utilization. World
Rev. Nutr. Dietetics, 22:152-182.
Orskov, E.R., 1977. Capacity for digestion and effects of composition of absorbed nutrients on
animal metabolism. J. Anim. Sci., 46: 600-608.
Orskov, E.R. and Allen, D.M., 1966a. Utilization of salts of volatile fatty acids by growing sheep.
1. Acetate, propionate and butyrate as sources of energy for young growing lambs. Br. J.
Nutr., 20: 295-305.
Orskov, E.R. and Allen, D.M., 1966b. Utilization of volatile fatty acids by growing sheep. 3.
Effect of frequency of feeding on the utilization of acetate and propionate by young growing
lambs. Br. J. Nutr., 20:509-517.
Orskov, E.R. and Allen, D.M., 1966c. Utilization of salts of volatile fatty acids by growing sheep.
4. Effects of type of fermentation of the basal diet on the utilization of salts of volatile fatty
acids for nitrogen retention and body gains. Br. J. Nutr., 20:519-532.
Orskov, E.R. and MacLeod, N.A., 1990. Dietary-induced thermogenesis and feed evaluation in
ruminants. Proc. Nutr. Soc., 49: 227-237.
Orskov, E.R., Hovell, F.D.DeB. and Allen, D.M., 1966. Utilization of salts of volatile fatty acids
by growing sheep. 2. Effect of stage of maturity and hormone implantation on the utilization
of volatile fatty acid salts as sources of energy for growth and fattening. Br. J. Nutr., 20: 307-
315.
Orskov, E.R., Flatt, W.P. and Moe, P.W., 1968. Fermentation balance approach to estimate
extent of fermentation and efficiency of volatile fatty acid formation in ruminants. J. Dairy
Sci., 51: 1429-1435.
(3rskov, E.R., Grubb, D.A., Smith, J.S., Webster, A.J.F. and Corrigal, W., 1979. Efficiency of
utilisation of volatile fatty acids for maintenance and energy retention by sheep. Br. J. Nutr.,
41: 541-551.
Owens, F.N. and Goetsch, A.L., 1986. Digesta passage and microbial protein synthesis. In: L.P.
Milligan, W.L. Grovum and A. Dobson (Editors), Control of Digestion and Metabolism in
Ruminants. Prentice-Hall, Englewood Cliffs, N J, pp. 196-226.
Pennington, R.J., 1952. The metabolism of short-chain fatty acids in the sheep. 1. Fatty acid
utilization and ketone body production by rumen epithelium and other tissues. Biochem. J.,
51: 251-258.
Pennington, R.J. and Pfander, W.H., 1957. The metabolism of short-chain fatty acids in sheep.
5. Some interrelationships in the metabolism of fatty acids and glucose by sheep-rumen epi-
thelial tissue. Biochem. J., 65:109-111.
Pennington, R.J. and Sutherland, T.M., 1956a. Ketone-body production from various sub-
strates by sheep-rumen epithelium. Biochem. J., 63: 353-361.
Pennington, R.J. and Sutherland, T.M., 1956b. The metabolism of short-chain fatty acids in the
sheep. 4. The pathway ofpropionate metabolism in rumen epithelial tissue. Biochem. J., 63:
618-628.
Perdok, H.B. and Leng, R.A., 1989. Rumen ammonia requirements for efficient digestion and
intake of straw by cattle. In: J.V. Nolan, R.A. Leng and D.I. Demeyer (Editors), The Roles
of Protozoa and Fungi in Ruminant Digestion. Penambul Books, Armidale, N.S.W., pp. 291-
293.
Peters, J.P., Leedle, J.A.Z. and Paulissen, J.B., 1989. Factors affecting the in vitro production
of volatile fatty acids by mixed bacterial populations from the bovine rumen. J. Anim. Sci.,
67: 1593-1602.
Peters, J.P., Shen, R.Y.W., Robinson, J.A. and Chester, S.T., 1990a. Disappearance and passage
of propionic acid from the rumen of the beef steer. J. Anim. Sci., 68: 3337-3349.
Peters, J.P., Shen, R.Y.W. and Chester, S.T., 1990b. Propionic acid disappearance from the
foregut and small intestine of the beef steer. J. Anim. Sci., 68:3905-3913.
Pethick, D.W., Lindsay, D.B., Barker, P.J. and Northrop, A.J., 1981. Acetate supply and utili-
zation by the tissue of sheep in vivo. Br. J. Nutr., 46:97-110.
Phillipson, A.T. and McAnally, R.A., 1942. Studies on the fate of carbohydrates in the rumen
of the sheep. J. Exp. Biol., 19: 199-214.
Poole, D.A. and Allen, D.M., 1970. Utilization of salts of volatile fatty acids by growing sheep.
5. Effects of type of fermentation of the basal diet on the utilization of salts of acetic acid for
body gains. Br. J. Nutr., 24: 695-704.
Preston, T.R., 1972. Molasses as an energy source for cattle. World Rev. Nutr. Dietetics, 17:
250-3 ! 1.
Preston, T.R. and Leng, R.A., 1987. Matching Ruminant Production Systems with Available
Resources in the Tropics and Subtropics. Penambul Books, Armidale, N.S.W., 245 pp.
M.F.Z van Houtert / Animal Feed Science and Technology 43 (1993) 189-225 223
Prins, R.A., 1977. Biochemical activities of gut micro-organisms. In: R.T.J. Clarke and T.
Bauchop (Editors), Microbial Ecology of the Gut. Academic Press, London, pp. 73-183.
Prior, R.L., 1978. Effect of level of feed intake on lactate and acetate metabolism and lipoge-
nesis in vivo in sheep. J. Nutr., 108: 926-935.
Reid, J.T., Ottilie, D., White, O.D., Anrique, R. and Fortin, A., 1980. Nutritional energetics of
livestock: some present boundaries of knowledge and future research needs. J. Anim. Sci.,
51: 1393-1415.
Reynolds, C.K., Tyrrell, H.F. and Reynolds, P.J., 1991. Effects of diet forage-to-concentrate
ratio and intake on energy metabolism in growing beef heifers: whole body energy and nitro-
gen balance and visceral heat production. J. Nutr., 121: 994-1003.
Rook, J.A.F., Balch, C.C., Campling, R.C. and Fisher, L.J., 1963. The utilization of acetic, pro-
pionic and butyric acids by growing heifers. Br. J. Nutr., 17: 399-406.
Rowe, J.B., Nolan, J.V. and Leng, R.A., 1978. Measurement of propionic acid and glucose me-
tabolism using a modelling approach. Proc. Aust. Soc. Anim. Prod., 12:136.
Russell, J.B., Strobel, H.J. and Martin, S.A., 1990. Strategies of nutrient transport by ruminal
bacteria. J. Dairy Sci., 73: 2996-3012.
Ryle, M. and Orskov, E.R., 1987. Rumen ciliates and tropical feeds. World Anim. Rev., 64: 21-
30.
Satter, L.D. and Slyter, L.L., 1974. Effect of ammonia concentration on rumen microbial pro-
tein production in vitro. Br. J. Nutr., 32: 199-208.
Scaife, J.R., Wahle, K.W.J. and Garton, G.A., 1978. Utilization of methylmalonate for the syn-
thesis of branched-chain fatty acids by preparations of chicken liver and sheep adipose tis-
sue. Biochem. J., 176: 799-804.
Schwartz, H.M. and Gilchrist, F.M.C., 1975. Microbial interactions with the diet and the host
animal. In: I.W. McDonald and A.C.I. Warner (Editors), Digestion and Metabolism in the
Ruminant. The University of New England, Armidale, N.S.W., pp. 165-179.
Smith, R.H., 1979. Synthesis of microbial nitrogen compounds in the rumen and their subse-
quent digestion. J. Anita. Sci., 49: 1604-1614.
Smith, R.M., 1971. Interactions of acetate, propionate and butyrate in sheep liver mitochon-
dria. Biochem. J., 124: 877-881.
Smith, S.B. and Crouse, J.D., 1984. Relative contributions of acetate, lactate and glucose to
lipogenesis in bovine intramuscular and subcutaneous adipose tissue. J. Nutr., 114: 792-
800.
Smith, S.B. and Prior, R.L., 1981. Evidence for a functional ATP-citrate lyase: NADP-malate
dehydrogenase pathway in bovine adipose tissue: Enzyme and metabolite levels. Arch.
Biochem. Biophys., 211: 192-201.
Smith, S.B. and Prior, R.L., 1986. Comparisons oflipogenesis and glucose metabolism between
ovine and bovine adipose tissues. J. Nutr., 116: 1279-1286.
Southon, S., Livesey, G., Gee, J.M. and Johnson, I.T., 1985. Differences in intestinal protein
synthesis and cellular proliferation in well-nourished rats consuming conventional labora-
tory diets. Br. J. Nutr., 53: 87-95.
Stangassinger, M. and Giesecke, D., 1986. Splanchnic metabolism of glucose and related energy
substrates. In: L.P. Milligan, W.L. Grovum and A. Dobson (Editors), Control of Digestion
and Metabolism in Ruminants. Prentice-Hall, Englewood Cliffs, N J, pp. 347-365.
Stern, M.D. and Hoover, W.H., 1979. Methods for determining and factors affecting rumen
microbial protein synthesis, a review. J. Anim. Sci., 49:1590-1603.
Stevens, C.E., 1970. Fatty acid transport through rumen epithelium. In: A.T. Phillipson (Edi-
tor), Physiology of Digestion and Metabolism in the Ruminant. Oriel Press, Newcastle-upon-
Tyne, UK, pp. 101-112.
Stevens, C.E., Argenzio, R.A. and Clemens, E.T., 1980. Microbial digestion: rumen versus large
intestine. In: Y. Ruckebusch and P. Thivend (Editors), Digestive Physiology and Metabo-
lism in Ruminants. MTP Press, Lancaster, UK, pp. 685-706.
Stewart, C.S., 1977. Factors affecting the cellulolytic activity ofrumen contents. Appl. Environ.
Microbiol., 33: 497-502.
Stouthamer, A.H. and Bettenhausen, C., 1973. Utilization of energy for growth and mainte-
nance in continuous and batch cultures of microorganisms. Biochim. Biophys. Acta, 391:
53-70.
Sutherland, T.M., 1977. The control and manipulation of rumen fermentation. In: D.J. Farrell
(Editor), Recent Advances in Animal Nutrition in Australia. The University of New Eng-
land, Armidale, N.S.W., pp. 110-129.
Tamminga, S., 1978. Measurement of microbial protein synthesis in the rumen. In: D.F. Os-
bourne, D.E. Beever and D.J. Thomson (Editors), Rumen Fermentation and Feed Evalua-
tion. Agricultural Research Council, London, pp. 5.1-5.11.
Tamminga, S., 1979. Relation between different carbohydrates and microbial synthesis of pro-
tein. Rep. No. 130, Institute for Livestock Feeding and Nutrition, Lelystad, 31 pp.
(Unpublished.)
Tappeiner, H., 1884. Untersuchungen fiber die G~irung der Cellulose insbesondere fiber deren
L~Ssungim Darmkanale. Zeitschr. Biol., 20: 52-134.
Ternouth, J.H. and Beattie, A.W., 1971. Studies of the food intake of sheep at a single meal. Br.
J. Nutr., 25: 153-164.
Thornton, R.F. and Tume, R.K., 1984. Fat deposition in ruminants. In: S.K. Baker, J.M. Gaw-
thorne, J.B. Mackintosh and D.B. Purser (Editors), Ruminant Physiology; Concepts and
Consequences. The University of Western Australia, Perth, pp. 289-298.
Thornton, R.F. and Tume, R.K., 1987. Factors controlling fat deposition in ruminants. In: D.J.
Farrell (Editor), Recent Advances in Animal Nutrition in Australia. The University of New
England, Armidale, N.S.W., pp. 28-49.
Tyrrell, H.F., Reynolds, P.J. and Moe, P.W., 1979. Effect of diet on partial efficiency of acetate
use for body tissue synthesis by mature cattle. J. Anita. Sci., 48: 598-606.
Ushida, K., Jouany, J.P. and Demeyer, D., 1991. Effects of presence or absence of rumen pro-
tozoa on the efficiency of utilization of concentrate and fibrous feeds. In: T. Tsuda, Y. Sasaki
and R. Kawashima (Editors), Physiological Aspects of Digestion and Metabolism in Ru-
minants. Academic Press, San Diego, CA, pp. 625-654.
Van der Walt, J.G., 1984. Metabolic interactions of lipogenic precursors in the ruminant. In:
F.M.C. Gilchrist and R.I. Mackie (Editors), Herbivore Nutrition in the Subtropics and
Tropics. The Science Press, Craighall, South Africa, pp. 571-593.
Van Nevel, C.J., Henderickx, H.K., Demeyer, D.I. and Martin, J., 1969. Effect of chloral hy-
drate on methane and propionic acid in the rumen. Appl. Microbiol., 17: 695-700.
Van Nevel, C.J., Demeyer, D.I. and Henderickx, H.K., 1971. Effect of fatty acid derivates on
rumen methane and propionate in vitro. Appl. Microbiol., 21: 365-366.
Van Soest, P.J., 1982. Nutritional Ecology of the Ruminant. O&B Books, Corvallis, OR, 374
PP.
Varnam, G.C., Jeacock, M.K. and Shepherd, D.A.L., 1978. Hepatic ketone-body metabolism in
developing sheep and pregnant ewes. Br. J. Nutr., 40: 359-367.
Veira, D.M., 1986. The role of ciliate protozoa in nutrition of the ruminant. J. Anim. Sci., 63:
1547-1560.
Vernon, R.G., 1981. Lipid metabolism in the adipose tissue of ruminant animals. In: W.W.
Christie (Editor), Lipid Metabolism in Ruminant Animals. Pergamon Press, Oxford, pp.
279-362.
Von Meting and Zuntz, N., 1877. In wiefern beeinflusst nahrungszufuhr die thierischen oxyda-
tionsprocesse. Pfluegers Archiv., 15: 634-636.
Wainman, F.W., Dewey, P.J.S. and Smith, J.S., 1982. The energy value to sheep of tallow and
of palm acid oil when added to a basal diet of barley and dried grass. In: A. Ekern and F.
Sundstol (Editors), Energy Metabolism in Farm Animals. The Agricultural University of
Norway, Aas-NLH, pp. 116-119.
Waldo, D.R., 1973. Extent of partition of cereal grain starch digestion in ruminants. J. Anim.
Sci., 37: 1062-1074.
Walker, D.J., 1965. Energy metabolism and rumen microorganisms. In: R.W. Dougherty (Edi-
tor), Physiology of Digestion in the Ruminant. Butterworths, Washington DC, pp. 296-3 t 0.
Warner, A.C.I., 1964. Production of volatile fatty acids in the rumen: methods of measurement.
Nutr. Abstr. Rev., 34: 339-352.
Webster, A.J.F., 1980. Energy costs of digestion and metabolism in the gut. In: Y. Ruckebusch
and P. Thivend (Editors), Digestive Physiology and Metabolism in Ruminants. MTP Press,
Lancaster, UK, pp. 469-484.
Webster, A.J.F., Osuji, P.O. and Weekes, T.E.C., 1976. Origins of the heat increment of feeding
in sheep. In: M. Vermorel (Editor), Energy Metabolism of Farm Animals. G. de Bussac,
Clermont-Ferrand, France, pp. 45-48.
Weekes, T.E.C. and Webster, A.J.F., 1975. Metabolism of propionate in the tissues of the sheep
gut. Br. J. Nutr., 33: 425-438.
Weigand, E., Young, J.W. and McGilliard, A.D., 1972a. Extent ofpropionate metabolism dur-
ing absorption from the ruminoreticulum. Biochem. J., 126:201-209.
Weigand, E., Young, J.W. and McGilliard, A.D., 1972b. Extent ofbutyrate metabolism by bo-
vine ruminoreticulum epithelium and the relationship to absorption rate. J. Dairy Sci., 55:
589-597.
Weller, R.A. and Pilgrim, A.F., 1974. Passage of protozoa and volatile fatty acids from the ru-
men of the sheep and from a continuous in vitro fermentation system. Br. J. Nutr., 32:341-
351.
Wolin, M.J., 1960. A theoretical rumen fermentation balance. J. Dairy Sci., 43: 1452-1459.
Wolin, M.J., 1975. Interactions between the bacterial species of the rumen. In: I.W. McDonald
and A.C.I. Warner (Editors), Digestion and Metabolism in the Ruminant. The University
of New England, Armidale, N.S.W., pp. 134-148.
Wolin, M.J. and Miller, T.L., 1983. Interactions of microbial populations in cellulose fermen-
tation. Fed. Proc., 42:109-113.
Yang, Y.T. and Baldwin, R.L., 1973. Lipolysis in isolated cow adipose cells. J. Dairy Sci., 56:
366-374.

Kelompok 4 VFA-1

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Kelompok 4 VFA-1

Uploaded by

Copyright:

Available Formats

Animal Feed Science and Technology, 43 ( 1993 ) 189-225 189

0377-8401/93/$06.00 © 1993 - Elsevier Science Publishers B.V. All rights reserved

The production and metabolism of volatile fatty

M.F.J. van Houtert ~

The energy associated with the phosphoanhydride bonds of adenosine tri-

Fermentation of carbohydrates to volatile fatty acids

Although ruminal fermentation of dietary proteins contributes to the pro-

Glucose- 6 - P XyluloseP RiboseP

~ 6 ~ ~ [ ~ F \ t ..... P [ <~1 Eryth.... P

3PGly...... 1~----11,3di-P-Glycerat~ I Aoo'o~oe','~oAF--~[ ~O~o~'r~' ]

2CH3 COCOOH + 4 [H ] + 4NADH + 2ADP

2CH3 COCOOH + 2 [ H ] + 2NADH + ADP

CO2 + 4H2 + ADP = CH 4 + 2H20 + ATP (5)

Fermentation of complex carbohydrates to pyruvate

Cellulose and hemicelluloses

nium and Polyplastron) also appear to possess ceUulolytic activity (Prins,

to constitute the major route of further hexose catabolism by the microorga-

Fermentation of pyruvate to volatile fatty acids

The central role of pyruvate in rumen fermentation continues, with the

Efficiency of microbial growth

Factors determining the efficiency of microbial growth are of great impor-

For maximal efficiency of microbial cell synthesis, it is desirable to stimu-

The extent of predation by protozoa on bacteria and the subsequent pref-

The nature, quantity and regularity in supply of fermentable substrate are

pacity and oxidation-reduction potential. Increases in liquid dilution rate

Absorption of volatile fatty acids from the rumen

Early in this century, the rumen epithelium was believed to be impermea-

croft et al. (1944) and McAnally (1944), however, showed unequivocally

Metabolism of volatile fatty acids in the rumen wall

Relatively little acetate is metabolised in the rumen epithelium (Penning-

Gluconeogenesis and other pathways of metabolism of volatile fatty acids in

Metabolism of acetate and butyrate

Propionate and gluconeogenesis

In roughage-fed ruminants, a large proportion of all the propionate which

circulation can increase significantly and propionate becomes available for

Other glucogenic precursors

Of the other glucogenic precursors, circulating amino acids, lactate and

Metabolism of volatile fatty acids in peripheral tissues and portal-drained

Metabolism of acetate in muscle tissue

Metabolism of acetate in adipose tissue

The biochemical reactions in fatty acid synthesis in ruminants have been

Metabolism of acetate in the portal-drained viscera

Utilisation of propionate and butyrate

While under normal conditions little propionate escapes metabolism in the

Energetic efficiency of utilisation of roughage based and concentrate-based

Roughage-based diets have been observed to be used with a lower energetic

Agricultural Research Council, 1980. The Nutrient Requirements of Ruminant Livestock.

You might also like