Maataloustieteellinen A ikakauskirja
Vol. 56: 265—274, 1984

Wood molasses as a preservative for high moisture barley.
2. Ration digestibility and rumen fermentation in sheep

PEKKA HUHTANEN

Department of Animal Husbandry, University of Helsinki,
SF-00710 HELSINKI 71, Finland

Abstract. The effect of wood molasses ensiled barley on ration digestibility and nitrogen
utilization (Exp. 1 and Exp. 2) and on rumen fermentation and degradation rate in sacco
(Exp. 2) was investigated in two experiments. The ration contained 0.9 kg of DM. The propor-
tion of hay was 30 % in Exp. 1 and 40 % in Exp. 2. In Exp. I the experimental diets were
dried barley (DB) and barley ensiled with a level of 8 (WMB8) or 16 % (WMBI6) wood mo-
lasses of barley DM. In Exp. 2 the diets were dried barley (DB), propionic acid-treated barley
(PAB) and barleys ensiled with 0.3 % v/w <7o of AIV II solution (AIVB) or with wood molas-
ses at a level of 12 % of barley DM (WMBI2).

The ration digestibility decreased with increasing levels of wood molasses. The difference
in DM and organic matter (OM) digestibility was significant (P < 0.05) between DB and
WMBI6. Wood molasses tended to decrease the apparent digestibility of crude protein and
crude fibre and to increase nitrogen retention. The percentages of nitrogen retained of
ingested were in Exp. 1 on DB, WMBB and WMBI6 diets 13.1, 17.8 and 15.8 % and in Exp.
2 on DB, PAB, AIVB and WMBI2 diets 13.8, 13.2, 10.3 and 14.5 %, respectively.

After feeding, the concentration of total VFA in the rumen was higher when ensiled
barleys were fed. The proportion of propionic acid in the total VFA was greater with DB and
PAB than with AIVB and WMBI2 diets (P < 0.01), and butyric acid was correspondingly
lower (P < 0.01 and P > 0.05). The proportion of isovaleric and valeric acids was highest
onAIVB diet. On DB and PAB diets the ammonia concentration in the rumen decreased after
feeding,but on AIVB and WMBI2 diets the highest value was reached 1.5 hours after feeding.
The degradation rate of DM and CP as determined by nylon bag method was faster on AIVB
and WMBI2 than on DB and PAB diets.

Introduction

Various types of wood molasses have been
fed to livestock, with a feeding value equal to
cane molasses (Turner 1964, Al-Chalabi et
al. 1974, CRAWFORoet al. 1978). Chang et al.

(1977) found no differences in diets con-
taining 8 and 12 % of spent sulphite liquor
and a control diet in beef cattle. Hartnell
and Satter (1978) suggested that the poly-
phenolic fraction in wood molasses (Maso-
nex) binds with certain proteins to decrease

Index words: grain preserving, wood molasses, sheep, digestibility, rumen fermentation.

265

JOURNAL OF AGRICULTURAL SCIENCE IN FINLAND

https://www.c-info.fi/en/info/?token=HGEEbdXcV5n9X7w0.dGVnP4cCxwUzukugFqf42Q.oAspxBBNjphkIrqSgZeFjCsAzewZDYO3zk8WAaGiIhVooYntkQbjcAXsvCX7fQ3rzV-v3fnhP2PfScONM9dFBrTm7ZDXNfuBWNR0KKA3f3aRvOlRZM6liVoveErXGriePouaGGbDyd43xsiMeWptF5G6PaZGpj0ZJUm63A


their microbiological degradation and in-
crease the amount of dietary nitrogen that
escapes the rumen fermentation.

Salo (1978) and Huhtanen (1984 a) re-
ported wood molasses, a by-product from
the wood processing industry, to be an effi-
cient preservative for high moisture barley.
The digestibility of barley ensiled with wood
molasses was lower than dried barley in pigs
(Huhtanen 1984 a) and in the diet of pigs
10—20 % wood molasses decreased the ra-
tion digestibility (Näsi 1984).

The objective of the present study was to
investigate the effects of wood molasses en-
siled barley on ration digestibility, nitrogen
utilization and rumen fermentation in sheep
and to evaluate the optimum level of wood
molasses in ensiled barley as an energy
source for ruminants. The wood molasses
ensiled barleys were the same used in the pre-
vious experiment (Huhtanen 1984 a).

Material and methods

The digestibility and rumen fermentation
trials were performed with three (Exp. 1) and
four (Exp. 2) Finnsheep rams in 3 X 3 and
4x4 Latin square arrangements. The aver-
age weight of the sheep was 48 kg in Exp. 1
and 55 kg in Exp. 2. The trials consisted of
transition, standardization and collection
periods lasting 5, 9 and 7 days each.

The sheep were kept in metabolism cages
allowing a separate collection of faeces and
urine. During the collection periods the
sheep were provided with faeces-collecting
harnesses.

In Exp. 1, diets comprised 300 g of hay
and either 700 g of dried barley (DB) or an
equal amount of DM of barley ensiled with
wood molasses at a level of 8 (WMB8) or
16 °7o (WMBI6) of barley DM; in Exp. 2
diets comprised 400 g of hay and 480 g DM
of dried barley (DB), propionic acid-treated
barley (PAB), barley ensiled with 0.3 °/o v/w
of AIV II solution (AIVB) or barley ensiled
with wood molasses at a level of 12 % barley
DM (WMBI2). In addition the animals re-

ceived a mineral mixture ad libitum in Exp.
1 and 20 g/d in Exp. 2. Water was given
freely and its consumption was measured
during the collection periods. Feeding took
place twice daily. The sheep were weighed
before and after the collection periods.

The faeces and urine were collected in the
morning and representative samples were
taken for analysis and stored frozen until
analyzed. DM determinations were made at
103°C; the samples for analysis were dried in
vacuum at 50°C for 2—3 days and milled
through a 1-mmscreen. The DM contents of
ensiled barleys were corrected according to
Jarl and Helleday (1948). The feed analy-
ses were made according to standard proce-
dures and VFA determinations by the meth-
od of Huida (1973).

During the two last days of collection peri-
ods in Exp. 2, rumen samples were taken
through the fistula before and 1.5, 3, 4.5 and
6 hours after the morning feeding. pH-mea-
surements were made immediately. The sam-
ples were centrifuged for 10 min at 2000
rpm. Ammonia N and VFA determinations
were made on the supernatant by the meth-
ods of McCullough (1967) and Huida
(1973). Samples for determination of rumen
microbiota were taken 6 hours after feeding.
Five ml of rumen content was transferred to
a glass bottle containing 45 ml of 10 % for-
malin. The total number of protozoa was
calculated according to Westerling (1970)
and the number of bacteria using a counting
chamber of dimensions 1 X 1 mm and 0.2
mm depth. Four preparations were made of
each sample for count.

The DM and crude protein (CP) degrad-
abilities of DB, PAB, AIVB and WMBI2
were determined in Exp. 2 by the method of
Setälä (1983). The experimental design was
4x4 Latin square and the incubation times
were 2,5, 9 and 24 hours. The degradability
measurements were performed on the same
barley fed in the diets.

The results were tested by analysis of vari-
ance and the differences between the means
by the Tukey-test.

266



Results and discussion

Digestibility of rations
The chemical composition of the experi-

mental feeds is presented in Table 1. Wood
molasses used at levels of 12 and 16 % de-
creased the digestibility of the ration (Ta-
ble 2). The differences in the DM and OM di-
gestibilities of DB and WMBI6 were signifi-
cant (P < 0.05). Also, the CP and crude
fibre digestibilities tended to decrease with
increasing levels of wood molasses. Ammo-
nium spent sulphite liquor (Salo and Puuma-
la 1978) and spent sulphite liquor (Chang et
al. 1977), which contain more lignosulphon-
ates than wood molasses have been found to

decrease the digestibility of forages and
soyabean-alfalfa substrate in vitro. In con-
trast, relative to cane molasses wood molas-
ses (Masonex) has been found to have posi-
tive effect on cellulose digestibility in steers
(Crawford et al. 1978). The wood molasses
used in the present experiment contains, how-
ever, more lignosulphonates than Masonex.

The effects of the various preserving meth-
ods on the ration digestibility agree with the
results of Weissbach and Schadereit (1968),
Korhonen et ai. (1973) and Ingalls et ai.
(1974). In contrast, Clark and Harsberger

(1972) obtained higher DM and OM digest-
ibilities on high moisture ensiled corn diet
than on dried corn diet. McKnight et al.

Table I. Chemical composition of experimental feeds

Exp. 1 Exp. 2
Hay DB WMBB WMBI6 Hay DB PAB AIVB WMBI2

Dry matter, % 84.5 87.5 57.2 56.0 80.9 87.0 83.0 55.0 54.9
In dry matter, %

Ash 7.7 3.0 4.0 5.2 5.9 2.7 2.8 3.2 3.5
Crude protein 10.2 11.8 12.1 11.0 8.9 11.6 13.7 12.7 12.1
Ether extract 1.7 2.2 2.6 2.3 2.2 2.3 2.5 2.8 2.7
Crude fibre 35.8 6.0 5.5 5.3 33.5 5.2 5.6 5.7 5.5
NFE 44.6 77.0 75.8 76.2 49.6 78.3 75.5 75.6 76.2

Table 2. Digestibility coefficients of total ration

Exp. 1 DB WMBB WMBI6

x s.d. x s.d. x s.d.

Dry matter 71.5a 3.1 70.7 ab 2.0 68.3b 2.6
Organic matter 73.5a 3.0 72.8“b 2.0 70.4b 2.3
Crude protein 68.1 2.3 67.8 2.1 63.1 2.3
Ether extract 70.3 4.1 77.9 1.9 78.3 1.0
Crude fibre 51.1 6.2 46.3 4.9 42.3 6.0
NFE 79.6 2.5 79.1 1.3 77.3 1.7
Exp. 2 DB PAB AIVB WMBI2

x s.d. x s.d. x s.d. x s.d.

Dry matter 71.3 2.3 72.1 3.2 71.9 0.4 69.8 2.2
Organic matter 73.2 2.2 74.1 3.1 73.7 0.5 72.0 2.0
Crude protein 67.4 1.9 73.0 2.4 69.8 3.0 68.4 4.4
Ether extract 72.9 3,4 75.2 1.0 76.8 4.6 76.7 5.0
Crude fibre 49.5 2.9 50.5 6.5 51.7 1.5 48.1 3.8
NFE 80.0 2.1 80.2 2.8 80.0 0.6 78.5 1.9

Means with different letters were significantly different: a, b (P < 0.05)

267



(1973) reported greater ruminal digestion of
DM, OM and starch for ensiled and acid-
treated corn diets than for dried corn diet
and this resulted in slightly better overall
digestibility of energy and DM. Likewise
Galyean et al. (1976) found better ruminal
starch digestion on ensiled corn than on
dried corn diet, but DM or OM digestion in
the rumen were not affected by the preserv-
ing method. The explanation for the higher
apparent CP digestibility of PAB in Exp. 2
could be the higher CP content of PAB.

Rumen fermentation
There were no significant differences in

the average pH-values or total VFA concen-
trations in the rumen (Table 3). At 1.5 and
3 hours after feeding, rumen pH was lower
and VFA concentration higher on the ensiled
barley diets than on DB or PAB diets
(Fig. 1). Similar effects of ensiled grain on
fermentation rate in the rumen have reported
by Torry and Perry (1974), Prigge et al.
(1976) and GALYEANet al. (1976) in vivo and
Danley and Vetter (1974) and Galyean et
al. (1976) in vitro.

The proportion of acetic acid was lower on
the PAB diet than on the other diets. The
difference was significant (P < 0.05)
between PAB and AIVB. Molar per cent of
propionic acid was higher (P < 0.01) on DP
and PAB diets than on ensiled barley diets,
and molar per cent of butyric acid lower
(P < 0.05) on DB than on AIVB or WMBI2
diets. Similar changes in VFA ratios when
ensiled grain is used have been reported by
Pratt and Conrad (1970), Ingalls et al.
(1974) and Tonrey and Perry (1974). Ga-
lyean et al. (1976), in contrast, reported a
lower acetate-propionate ratio when ensiled
corn was fed to steers instead of dried corn.
The slightly lower acetate-propionate ratio
on the PAB diet than on the DB diet agrees
with the results of Clark et al. (1973) and
may be due to the prior treatment of barley
with propionic acid.

The high proportion of butyric acid on
ensiled barley diets may partly be explained
by higher sugar content of ensiled barley
than of dried barley. Increased levels of mo-
lasses in the diet have been found to promote
butyrate production (Karalazos and Swan

Table 3. Rumen pH, ammonia N, VFA and microbes on experimental diets. The values are averages of samples
taken at different times.

DB PAB AIVB WMBI2

x s.d. x s.d. x s.d. x 7 s.d.

pH 5.97 0.32 5.97 0.31 5.90 0.36 5.86 0.28
Ammonia N mmol/1 8.1 a 5.2 12.3ab 2.6 11.lab 3.9 12.7b 5.9
Total VFA mmol/1 81.2 10.0 79.7 6.5 82.5 13.9 85.3 11.3
Molar per cent of

Acetic acid 59.4ab 1.8 57.l b 2.1 60.2a 2.9 59.1 ,b 4.6
Propionic acid 25.9d 3.1 26.6 d 3.3 20.7' 3.2 22.2' 2.5
Butyric acid 11.0' 3.0 13.011' 1.8 14.ld 2.0 14.7d 2.4
Isovaleric acid 2.3d 0.7 1.8d 0.4 2.9' 1.0 2.1 d 0.3
Valeric acid 1.3d 0.3 1.5"' 0.2 2.0r 0.7 1.7' f 0.5
Caproic acid 0.0“ 0.0 0.0s 0.0 0.1 ab 0.1 0.2 b 0.2

Ratio A:P 2.3' f 0.3 2.2 f 0.4 3.0“ 0.5 2.7 d' 0.5
Ratio A:B 5.911 2.1 4.5d' 0.7 4.4d ' 0.8 4.2' 1.1
Ratio P:B 2.6a 1.0 2.1 ab 0.5 1.9b 0.9 1.8» 0.6
Bacteria n x 109 ') 4.67a 4.00ab 2.84ab 2.16 b

Protozoa n x 104 ') 87.4 115.2 114.9 90.3

‘) 6 hours after feeding
Means with different letters significantly different: a, b, c (P < 0.05), c, d, f (P < 0.01)

268



269

Fig. I. Rumen pH, ammonia N and VFA of sheep fed with different barleys (A DB, APAB, o AIVB and
• WMBI2)



1974). Syrjälä (1972) reported higher pro-
portion of butyrate when sucrose was added
to a grass silage diet instead of starch. But
also a change in the ratio of the number of
bacteria and protozoa has an effect on the
propionate-butyrate ratio. Increases in cili-
ate number are proportionate to decrease in
propionic acid and increase in butyric acid
(Eadiecl al. 1970, Whitelaw et al. 1972). In
the present experiment the ratio between the
number of bacteria and protozoa tended to
be lower on ensiled barley diets (Table 3).
IsHAQUEet al. (1971) reported that a change
in fermentation pattern from propionate to
butyrate increases the proportion of OM di-
gested in the rumen.

The higher (P < 0.01) proportion of iso-
valeric acid on AIVB diet may be attributed
to decreased utilization or increased deamina-
tion by rumen micro-organisms (Bruggeman
and Gieseke 1967). The lower proportion of
isovaleric acid on WMBI2 diet than on
AIVB diet is in agreement with in vitro re-
sults reported by Chalupaand Montgomery

(1979). At lower levels wood molasses was
fermented to methane, acetate and butyrate,
whereas increased supplement level caused
increased production of propionate (Chalu-
pa and Montgomery 1979).

On ensiled barley diets, rumen ammonia N
concentration reached a peak 1.5 hours after
feeding, but on DB and PAB diets the
highest concentration was found before feed-
ing. On average, ammonia N concentration
was higher (P < 0.05) on WMBI2 diet than
on DB diet. A higher rumen ammonia con-
centration with ensiled grains has also been
reported by McKnight et al. (1973) and
PRiGGEet al. (1978). In contrast, Ingalls et
al. (1974) and Prigge et al. (1976) found
lower rumen ammonia level on ensiled than
on dried grain diets. One explanation for the
higher ammonia level after feeding may be
increased protein solubility during ensiling.
The positive relationship between protein
solubility and ammonia release in the rumen
has been demonstrated in many experiments
(Annison et al. 1956, Donaldson and
Edwards 1977). A reason for the different
shapes of ammonia curves may be the change
in rumen microbiota. It has been shown that
an increase in the number of protozoa and
decrease in the number bacteria increases
rumen ammonia concentration (Klopfen-
stein et al. 1966 Males and Purser 1970,
Veiracl al. 1983). The correlations between
rumen ammonia concentration and VFA
ratios in the rumen were in good agreement

Fig. 2. DM and CP disappearance of different barleys in the rumen ( A DB, A PAB, o AIVB and • WMBI2)

270



with those observed by Males and Purser
(1970).

Degradability of DM and CP

DM and CP degradation rates in the
rumen were faster for ensiled barleys than
for DB and PAB (Fig. 2) which is in agree-
ment with the rumen fermentation studies
and in sacco studies by Galyeancl al. (1977)
with ensiled corn. The difference in the de-
gradation rates of DM and CP decreased
with longer incubation times, and after an
incubation time of 24 hours the DM degrad-
ability of DB was higher (P < 0.05) than to
WMBI2. Dried barley had a slower degrada-
tion rate than reported earlier by Setälä
(1983) and the values were more similar to
those reported by Lindberg and Varvikko
(1982).

Nitrogen utilization

In Exp. 1 the use of wood molasses as a
preservative tended to increase nitrogen re-
tention in sheep aged B—lo months, com-
pared with DB (Table 4). The excretion of
nitrogen in urine was lower (P < 0.05) and
in faeces higher (P < 0.05) on WMBI6 than
on DB diet. Blood urea N was not deter-
mined, but was lower (P < 0.01) in growing
bulls fed on WMB 12 diet than in bulls fed on
AIVB diet (Huhtanen 1984 b). A reason for
the slightly better protein utilization on
WMB diets is suggested by the finding that

xylose is a better energy source for microbial
protein synthesis in vitro than sucrose or
mannose (Henderickx and Martin 1963).

The increased protein solubility of the en-
siled barleys had no adverse effect on protein
utilization. After feeding of ensiled barley
diets microbes had more rapidly fermenting
organic matter available to use soluble nitro-
gen for protein synthesis. Generally, proteins
of low solubility are used more efficiently
than proteins of high solubility because more
dietary protein reaches the duodenum intact
and less cycles through the process of de-
gradation to ammonia. However, Prigge et
al. (1976) observed greater nitrogen retention
with high moisture ensiled corn than with
dried rolled corn. Prigge et al. (1978) re-
ported abomasal nitrogen flow and efficiency
of microbial synthesis to be greater on ensiled
or acid-treated corn than on dried corn. The
increased microbial synthesis was related to
increased rumen turnover rate. Increased
rumen dilution rate has been found to be as-
sociated with increased acetate and butyrate
and decreased propionate in rumen VFA
(Thomson et al. 1975, Owens and Isaacson
1977). This effect was also found on the
ensiled barley diets of the present study.

When dietary protein is of poor quality,
utilization can be improved by rumen de-
gradation and conversion to microbial pro-
tein (Little et al. 1963, Prigge et al. 1976).
Moreover, when the grain protein is pro-
tected against microbial degradation in the
rumen, no positive effect on protein utiliza-

Table 4. Nitrogen balance and excretion of nitrogen in faeces and urine.

Exp. 1 Exp. 2
DB WMBB WMBI6 DB PAB AIVB WMBI2

N intake, g/d 15.7 16.3 15.0 13.5* 15.4b 14.8>b 13.7*
N in faeces, g/d 5.0s SJ» 11 5.5 b 4.4 4.2 4.5 4.3
N in urine, g/d 8.8» 8.1» b 7.1» 7.2 9.2 8.9 7.4
N balance, g/d 2.1 2.9 2.6 1.9 2.0 1.5 2.0
% of intake 13.1 17.8 15.8 13.8 13.2 10.3 14.5
% of absorbed 19.6 26.4 25.3 20.6 18.2 14.6 20.4 -

Means with different letters significantly different: a, b (P < 0.05)

3 271



tion is found (Davis and Faichney 1973,
Thornton et al. 1977). In contrast to the
results of Hartnell and Satter (1978), in
the present experiment wood molasses did
not slow the degradation of protein by rumen
microbes. But it did decrease the breakdown
of barley protein in the silo compared with
AIV II solution (Huhtanen 1984 a).

When wood molasses was used as a preser-
vative for high moisture barley at a level of

B—l 2 % of barley DM, no significant effect
on ration digestibility was found and it
tended to improve protein utilization. Wood
molasses at this level can therefore serve as
an energy source as well as a preservative for
grain.

Acknowledgements. I wish to express my thanks to
Mr. Matti Järvi for taking care of the animals and for
technical help.

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273



SELOSTUS

Puumelassi tuoreen ohran säilöntäaineena.
2. Vaikutus rehun sulavuuteen ja pötsi-
fermentaatioon lampaalla

Pekka Huhtanen
Helsingin yliopisto', kolieläintieteen laitos,
00710 Helsinki 71

Kahdessa kokeessa selvitettiin puumelassilla säilötyn
ohran vaikutusta rehuannoksen sulavuuteen ja typen
hyväksikäyttöön (koe 1 ja 2). Kokeessa 2 selvitettiin li-
säksi vaikutusta pötsifermentaatioon ja rehun hajoa-
misnopeuteenpötsissä. Koe-eläimet saivat kuiva-ainetta
0.9 kg/pv, josta heinän osuus oli kokeessa 1 30 % jako-
keessa 2 40 %. Koerehuina oli kokeessa 1 kuivattu
(KO), sekä 8 (PM08) ja 16 fo:n puuraelassilisäyksellä
säilötty ohra (PMOI6) sekä kokeessa 2 kuivattu (KO),
propionihapolla jyväsäilötty (PrO), AIV II;lla (AIV2O)
tai puumelassilla (12 % ohran kaista) säilötty ohra
(PMOI2).

Puumelassitason noustessa rehuannoksen sulavuus
huononi. Ero oli merkitsevä (P < 0.05) KO:n ja
PMO!6:n kuiva-aineen ja orgaanisen aineen sulavuu-
dessa. Puumelassilla säilötty ohra huononsi hieman raa-
kavalkuaisen ja raakakuidun näennäistä sulavuutta ja
lisäsi typen pidättymistä (P > 0.05). Pidättyneen typen

osuus typen saannista oli kokeessa 1 KO-, PMOB- ja
PMO!6-ruokinnalla 13.1, 17.8 ja 15.8 % sekä kokeessa
2 KO-, PrO-, AIV2O- ja PMOI2- ruokinnalla 13.8,
13.2, 10.3 ja 14.5 % vastaavasti.

KO- ja PrO-ruokinnalla pötsinesteen pH laski ja
VFA-pitoisuus nousi vähemmän kuin murskesäilötyillä
viljoilla. Propionihapon osuus VFA:sta oli KO- jaPrO-
ruokinnalla korkeampi (P < 0.01) ja voihapon osuus
KO-ruokinnalla alempi (P < 0.01) kuin AIV2O- ja
PMOI2-ruokinnalla, Murskesäilöttyä ohraa käytettäes-
sä pitkäketjuisten rasvahappojen osuus VFA;sta lisään-
tyi. KO- ja PrO-ruokinnalla pötsinesteen ammoniak-
kipitoisuus alkoi laskea ruokinnan jälkeen, mutta
AIV2O- ja PMO!2-ruokinnalla ammoniakkihuippu saa-
vutettiin 1.5 tuntia ruokinnan jälkeen. Murskesäilöntä
lisäsi sekä kuiva-aineen että raakavalkuaisen hajoamis-
nopeutta pötsissä nailonpussimenetelmällä määritetty-
nä.

274