THE DIGESTION OF CELLULOSE BY SHEEP
THE EXTENT OF CELLULOSE DIGESTION AT SUCCESSIVE
LEVELS OF THE ALIMENTARY TRACT
BY F. V. GRAY
From the Division of Biochemistry and General Nutrition of the Council for
Scientific and Industrial Research, University of Adelaide, South Australia
{Received 4 October 1946)
Until recently, information of the extent to which cellulose is digested in the
alimentary tract of the ruminant was available only indirectly through studies of the
fate of that indefinite and variable complex mixture of cellulose, hemicellulose and
encrusting substances which is conventionally referred to as 'crude fibre'. This,
in the past, has been taken as a measure of the less readily digested part of the
carbohydrates of plants, in contrast to the ' N-free extractives' which represent the
more soluble and therefore more readily digestible portion.
Popoff (1875) observed the formation of methane in the rumen as a product of the
fermentation of crude fibre; Zuntz described similar fermentative changes in the
caecum, and Hofmeister (1880) showed that they also occurred in the colon.
Tappeiner (1882) attempted to assess the extent of digestion of crude fibre of the
food at different levels as it passed along the alimentary tract of the ruminant. He
took samples of the contents of the rumen, small intestine, and caecum from a
freshly killed sheep, and estimated the crudefibrewhich disappeared after' complete'
fermentation at body temperature. From this he concluded that the greater part of
the crude fibre was digested in the rumen, a smaller amount in the caecum, and
none at all in the small intestine.
A number of workers (Williams & Olmstead, 1935; Norman, 1935; Crampton &
Maynard, 1938) have stressed the serious limitations of conventional analyses which
divide the carbohydrates of plant foodstuffs into crude fibre and N-free extractives,
and have suggested that the analysis of such foods would be more logical and
informative if the carbohydrates were divided into better defined groups, such as
cellulose, lignin, and other carbohydrates. This proposal has been adopted and
extended by Hale, Duncan & Huffman (1940) in their study of digestion in the
bovine rumen. These authors developed a method for estimating the extent of
digestion in the rumen by the use of ' lignin ratios' and concluded that 85 % of the
digestible cellulose in alfalfa hay was broken down there.
The following experiments were carried out in this laboratory during 1936. They
were mentioned in the Annual Report of this laboratory (Commonwealth of
Australia, Council for Scientific and Industrial Research, 1937-38).
16
F. V. GRAY
METHOD
The lignin ratio procedure was used to determine the extent of cellulose digestion at
successive levels of the alimentary tract of the sheep. The method is based essentially
on a procedure suggested by Bergeim (1926) for the determination of digestion
coefficients. After adding Fe3O4 to the food Bergeim determined the ratio of the
amount of a given nutrient to the amount of iron in the food and in the faeces, and
from this the percentage utilization was calculated. Later Heller, Breedlove &
Likely (1928) used the natural iron content of the food as a reference substance, to
ensure a more uniform distribution; Gallup (1929) and Gallup & Kuhlman (1931)
made use of silica for the same purpose.
The validity of such procedures depends fundamentally on the complete indigestibility of the reference substance, and the exact concurrence of its passage
through the alimentary tract with that of the nutrient in question. Knott, Murer &
Hodgsen (1936) came to the conclusion that added materials, such as iron oxide,
were unsuitable for the calculation of digestion coefficients in ruminants because
they were unlikely either to remain evenly distributed in the fodder, or pass along
the tract at the same rate as the rest of the food. For a similar reason silica was
considered also to be unsuitable; although it is a natural and intimate constituent of
plant structures, sand is nearly always present in fodders and may gather in certain
parts of the tract rather than move along it at an identical rate with the rest of the
food. The absorption of part of the natural iron of the fodder debars its use as
a critical reference marker.
Hale et al. (1940) have employed lignin as a reference substance for the calculation
of digestion coefficients in the rumen. They claimed that it was digestible to a
variable extent (up to 24%) in the lower levels of the tract. There have been other
claims that lignin is not undigested in various animals including the ruminant (cf.
Konig, 1907; Csonka, Phillips & Jones, 1929; McAnally, 1942), and there are others
to the contrary (Paloheimo, 1925; Rogozinski & Starewska, 1927; Crampton &
Maynard, 1938). As lignin is not a chemical entity the term, of necessity, has often
been used loosely to signify a mixture of similar complexes, some of which may be
prone to change in the alimentary tract, and thus may not be expected to react
identically towards the various reagents and procedures which different authors
have used for its estimation. Since the different methods employed are all empirical,
the origin of the controversy is clear enough. Norman (1935) directed attention to
this and pointed out that experiments which have depended on the fate of isolated
lignin added to the diet are not conclusive since the biological availability of lignin
is affected by the processes of isolation.
Lignin was used as a reference marker in the following experiments after it was
proved that, as determined in a mixture of wheat straw and lucerne hay by the
method of Norman & Jenkins (1934), it was not digested to any appreciable extent
by sheep.
The digestion of cellulose by sheep
17
EXPERIMENTAL
(1) The digestion of lignin by sheep
The evidence that lignin is not digested was drawn from the study of the intake and
output of lignin by three sheep fed on a constant diet of wheat straw and lucerne
hay for several months while confined to metabolism cages described by Marston
(1935). During the fourteen day collection periods the fodder eaten and the faeces
voided were measured with meticulous care. Lignin was determined by the method
of Norman & Jenkins (1934) in samples from each and the balance drawn up. The
data are summarized in Table 1.
Table 1. Digestion of lignin by sheep
Sheep
no.
Dry wt.
fodder
eaten
(g-)
Dry wt.
faeces
voided
(g.)
Fodder
lignin
(%)
Faeces
lignin
(%)
Lignin
eaten
(g-)
Lignin
voided
(g.)
WaA3
W 3 A 43
W 0 22O
W 6 22O
W 8 22O
7616
753O
7620
7480
8130
4308
4343
4325
4184
4813
13-84
1204
12-40
12-43
1270
24-72
2091
20-92
2200
2101
1050
907
945
93O.
1030
1060
908
905
920
IOIO
It may be noted that these results are in agreement with those of Crampton &
Maynard (1938) for the digestion of lignin by steers. A different method was used
for the determination of lignin, and in which different fodders were fed.
(2) Extent of cellulose digestion at successive levels of the alimentary tract
Four sheep which had been fed for some weeks previously on a constant ration
composed of wheat straw and lucerne hay were slaughtered, and samples of ingesta
were taken from the rumen, omasum, abomasum, small intestine, caecum and
colon. The dried, ground samples were subjected to analyses for cellulose and
lignin. The method for the determination of lignin has been referred to previously;
the method used for the determination of cellulose was that recommended by
Norman & Jenkins (1933).
Similar analyses were made of samples of the fodder which the animals had been
fed.
Results of these analyses are listed in Table 2.
The data indicate that, of the cellulose present in the fodder, 40-45 % was digested
before the food passed into the abomasum, and an additional 15-20 % was digested
in the large intestine. During this second fermentation 7-11 % disappeared in the
caecum, and 4-9 % in the colon. No appreciable loss of cellulose occurred in the
abomasum or in the small intestine.
Since the total digestion of cellulose amounted to about 60%, it is apparent that
approximately 70% of the cellulose that was digested was broken down in the
rumen, 17% in the caecum and 13% in the colon.
j EB.24, i & 2
2
i8
F. V. GRAY
Table 2. Extent of cellulose digestion at successive levels of
the alimentary tract
Level
Fodder
Rumen
Omasum
Abomasum
Small intestine
Caecum
Colon
Fodder
Rumen
Omasum
Abomasum
Small intestine
Caecum
Colon
Fodder
Rumen
Omasum
Abomasum
Small intestine
Caecum
Colon
Fodder
Rumen
Omasum
Abomasum
Small intestine
Caecum
Colon
% lignin
% cellulose
10-41
17-20
16-81
1379
1090
15-12
18-91
3649
3564
3S63
12-06
20-71
1907
1689
13-11
2O-2O
19-79
38-61
39-96
28-40
220
2661
29-01
41-60
3241
2450
33-n
27-76
Ratio
3'5°
2-07
2-12
2-06
2 O2
176
i-53
3-20
i-93
2-18
1 92
1-87
1-64
1-40
3861
4109
37-72
2949
2586
29-20
26-50
3 20
1-91
i3 - 94
1998
38-48
276
3401
1-70
14-21
1401
1948
20-76
2245
22-59
26-70
23-06
158
161
12-06
21-50
21-24
17-35
i5-4i
22-14
22-30
1-78
1-70
i-68
1-32
1-19
i-37
i-ii
% cellulose
digested
—
408
39-4
412
42-3
497
562
—
397
31-9
40-0
41-6
48-7
56-4
—
4°-5
44-4
46-8
47-4
58-7
628
—
384
42-8
41-7
50-4
59-8
The claims of Hofmeister (1880) and Tappeiner (1882, 1884) that some digestion
of cellulose occurs in the caecum and colon of ruminants were supported by the
work of the Dresden school (Ellenberger, 1915-16), although it was stated that the
quantity that disappeared in the colon was small. Trautmann & Asher (1939),
however, found no fermentation of chemically pure cellulose which had been
introduced directly into the caecum and colon of a goat, even after a prolonged
sojourn there. They observed no loss of cellulose when rumen contents were
introduced into the caecum, and after 24 hr. there, they considered it to be free of
rumen flora. These workers claimed that the time ingesta stay in the caecum and
colon, together with the fact that there is a rapid water resorption from these organs,
must be inhibitive to fermentation processes.
The present experiments, however, support the contention of the earlier investigators that this portion of the tract plays a significant part in the digestion of
cellulose by the sheep. Indirect evidence which further supports this view is to be
seen in the work of Barcroft, McAnally & Phillipson (1944) who found that volatile
fatty acids, the acknowledged products of cellulose fermentation, are present in the
blood draining the caecum in considerably greater quantity than in the blood of the
peripheral circulation.
The digestion of cellulose by sheep
19
SUMMARY
1. Lignin, as determined in these experiments, has been shown to be undigested
by sheep.
2. Examination of cellulose-lignin ratios in the fodder, and in material taken
from successive levels of the alimentary tracts of sheep indicated that 70 % of the
digestible cellulose of the fodder was broken down in the rumen, 17% in the
caecum, and 13 % in the colon. No digestion of cellulose occurred in the abomasum,
or in the small intestine.
The author is indebted to Mr H. R. Marston, Chief of the Division, for suggesting
the problem, for the specimens from which the lignin balances were determined,
and for his help and criticism throughout.
REFERENCES
BARCROFT, J., MCANALLY, R. A. & PHILLIPSON, A. T. (1944). J. Exp. Biol. 20, 120.
BERGEIM, O. (1926). J. Biol. Chem. 70, 29.
COMMONWEALTH OF AUSTRALIA, COUN. SCI. INDUSTR. RES. (1937-38). 12th Ann. Rep. p. 38.
CRAMPTON, E. W. & MAYNARD, L. A. (1938). J. Nutrit. 15, 383.
CSONKA, F. A., PHILLIPS, M. & JONES, D. B. (1929). J. Biol. Chem. 85, 65.
ELLENBERGER, W. (1915-16). Hoppe-Seyl. Z. 96, 236.
GALLUP, W. D. (1929). J. Biol. Chem. 81, 321.
GALLUP, W. D. & KUHLMAN, A. H. (1931). J. Agric. Res. 42, 665.
HALE, E. B., DUNCAN, E. W. & HUFFMAN, C. F. (1940). J. Dairy Sci. 23, 953.
HELLER, V. G., BREEDLOVE, C. H. & LIKELY, W. (1928). J. Biol. Chem. 79, 275.
HOFMEISTER, V. (1880). Arch. wiss. u. prakt. Tierheilk. 7, 169.
KNOTT, J. C , MURER, H. K. & HODGSEN, R. E. (1936). J. Agric. Res. 53, 5^3.
KONIG, J. (1907). Landw. VerSta. 65, 55.
MARSTON, H. R. (1935). J. Agric. Sci. 25, 103.
MCANALLY, R. A. (1942). Biochem. J. 36, 392.
NORMAN, A. G. (1935). J. Agric. Sci. 25, 529.
NORMAN, A. G. & JENKINS, S. H. (1933). Biochem. J. 27, 818.
NORMAN, A. G. & JENKINS, S. H. (i934)- Biochem. J. 28, 2147, 2160.
PALOHEIMO, L. (1925). Biochem. Z. 165, 463.
POPOFF, L. (1875). Pfliig. Arch. ges. Physiol. 10, 113.
ROGOZINSKI, F. & STAREWSKA, M. (1927). Intern. Rev. Agric. 18, 413.
TAPPEINER, H. (1882). Ber. dtsch. chem. Ges. 15, 999.
TAPPEINER, H. (1884). Z. Biol. 20, 52.
TRAUTMANN, A. & ASHER, T. (1939). Z. Tier. Futtermitt. 3, 45.
WILLIAMS, R. D. & OLMSTEAD, W. H. (1935)- J- Biol. Chem. 108, 653.