Professional Documents
Culture Documents
Effect of
supplementation with Gliricidia sepium and
urea/molasses
Héctor Osorio De La Cruz
Abstract
Introduction
Cellulose is the world's most widely available renewable resource, amounting to
about fifty per cent of the cell-wall material of woody and herbaceous plants.
Due to this abundance and renewabili- ty, there has been a great deal of interest
in utilizing cellulose as an energy resource and as a feedstock (Fan et al 1982).
Many fibrous by-products have a substantial potential value as animal feedstuffs.
Ruminants, especially, have the unique capacity to utilize cellulose, because of
their microbes (Boucqué and Fiems 1988). The shortage and high costs of
conventional raw materials necessitates the use of these agricultural residues in
animal feeds. However, the high lignin content, the low level of soluble
carbohydrate and the relative absence of both fermentable nitrogen and bypass
protein are responsible for the low nutritional value of untreated residues (Hamad
and El-Saied 1982; Preston and Leng 1974; Sundstol 1988).
Agricultural by-products like cereal straws and sugar-cane bagasse are high in
ligno-cellulose. Roughly three-quarters of straw is cellulose plus hemicellulose.
Bagasse also contains more than 60% of its dry matter in the form of cellulose
and hemicellulose but its degradability is very poor. One of the main reasons for
this depression in degradability is the presence of lignin which protects
carbohydrates from attack by the rumen microbes. Sugar- cane bagasse contains
around 50% cellulose, 27.9% hemicellulose, 9.8% lignin and 11.3% cell contents
(Kewalramani et al 1988).
It has been recognized that in order to improve the nutritive value of ligno-
cellulosic materials for livestock, some form of pretreatment of processing of the
plant material is required (Helmling et al 1989).
Polysaccharide degradation in the rumen takes place as the result of the action of
a consortium of anaerobic bacteria, protozoa and phycomycete fungi and, even
so, its bioconversion is far from complete. Hence a pretreatment of the substrate
is required to increase the rate and extent of holocellulose hydrolysis, in order to
alter significantly the structural characteristics of the ligno- cellulosic matrix.
Such a pre-treatment must enhance the close contact between microbe and fibres
to provide an efficient enzyme action (Rolz et al 1987).
A technique which has shown considerable potential for the cost- effective pre-
treatment of ligno-cellulosic materials is that of steam explosion (Wong et
al 1974). With this method, the substrate is loaded into a pressure vessel and
heated by steam injection for a defined time-temperature period. At the end of
this period, the contents are explosively discharged into a pressure vessel; a
substantial portion of the hemicellulose fraction is made water soluble and the
lignin fraction is modified (Wang et al 1974, Morjanoff and Gray 1987).
The most noticeable effect of treating raw bagasse with steam under pressure is a
large reduction in certain fibre components. Crude fibre was reduced from 43 to
34% and NDF was reduced from 83 to 51%; however, there was little change in
ADF, cellulose and lignin content. These data indicate that steam-pressure
treatment com- pletely modified the hemicellulose fraction of raw bagasse, chan-
ging it into more soluble components, but did not affect the ligno- cellulose
components (Wong et al 1974; Pate 1982; Kling et al 1987)
When sugar-cane bagasse (and fibrous residues in general) is used as the basal
diet, it is important to give the correct supplementa- tion in order to obtain
satisfactory physical and economic respon- ses. The supplementation must take
account of the productive stage of the animals (eg: growing, fattening, lactating,
etc) and the locally available sources of supplements in the region (Preston and
Leng 1987). The nutrients arising from rumen fermentation must be balanced
using supplements containing dietary by-pass protein to provide essential amino-
acids, dietary by-pass starch which provide extra glucose, and dietary fat to
provide long chain fatty acids for the synthesis of body tissues and milk fat. The
low N content of sugar-cane and its by-products, impairs rumen fermetation due
to the low availability of ammonia and sulphur, in animals fed on these diets
(Leng 1989).
Gliricidia sepium (Jacq.) Steud, is a legume tree which has been used for a long
time in Colombia and other countries, mainly as a live fence. It is a tropical
species which grows at altitudes from 0 to 1500 metres above sea level (Baggio
1982). The foliage of Gliricidia sepium is a combined source of micro-nutrients
for the rumen bacteria, vitamin A and some fermentable and by-pass protein to
balance the energy from the rest of the diet. Moreover, it also provides long fibre,
which is needed in diets in which bagasse is the basal diet. Diets based on
bagasse are deficient in long chain fatty acids; supplementation with rice bran
provides a suitable supply and also gives additional by-pass nutrients in the form
of starch, lipids and protein (Elliot et al 1978). Poultry litter is a local and natural
resource, rich in macro-(Ca, P, S) and micro- minerals (Cu, Co, Zn, etc), because
of the routine supplementation with balanced minerals in intensive poultry
enterprises; it also provides fermentable nitrogen, mostly as uric acid which is
hydro- lysed to ammonia by rumen micro-organisms (Preston and Leng 1984).
Urea supplementation increases the utilisation of low-nitrogen fibrous diets. A
liquid mixture of urea/molasses is attractive and palatable to ruminants because
of the smell and taste of molasses; the animals lick it almost continuously, and as
a result its ingre- dients are continuously available to the rumen micro-organisms.
Although chemical analysis and IVOMD are valid first steps in assessing the
relative values of chemical pre-treatments, responses may be modified in the in
vivo situation. For example, although ammonia treatments increase the nitrogen
contents of by-products, the availability of the retained nitrogen to the animal
may not be high (Ibrahim and Pearce 1983a).
Objectives
The purpose of this experiment was to find out if the physical treatment (steam)
and the combination of a physical and a chemical treatment (steam-ammonia) of
sugar cane bagasse were suitable treatments for this sugar industry by-product, in
order to use it later as an animal feed. The optimum levels of supplementation
with foliage of a legume tree (Gliricidia sepium), and urea/molasses liquid
mixture were also tested. In vivo degradability tests were made to obtain
additional information about the effect of the treatments on the sugar cane
bagasse, and the effect of the supplementation on the degradability of the sugar
cane bagasse.
Experiments were carried out at sites adjacent to the Ingenio del Cauca and
Ingenio Providencia, sugar factories, both situated in the Department of Valle del
Cauca, Colombia (4°N, 76°W, 957 metres above sea level, mean temperature
24°C).
A first experiment was carried out to study the effect on live weight change in
Zebu cattle (Bos indicus) of steam-treated sugar- cane bagasse supplemented
with two different levels of a legume tree foliage (Gliricidia sepium) and four
quantities of an urea/molasses liquid mixture which contained 10% of urea.
On the basis of the results obtained in that first experiment, a second one was
designed in which lower levels of Gliricidia sepium and three different
percentages of urea in the molasses/urea liquid mixture were used, and
measurements were made of the live weight change of Zebu cattle. In this second
experiment a comparison was also made between bagasse treated with steam (as
in the first one) vs. steam and ammonia treatment and their effects on the live
weight change of Zebu cattle.
In vivo degradability tests by the nylon bag method (Orskov et al 1980) were
carried out in Experiment 3, using animals of the same type as those in the
previous two experiments, mainly to assess the effect of the treatments on the
degradability of bagasse, and the effect, if any, of supplementation
with Gliricidia sepium and poultry litter.
Experimental animals
The animals used in all the experiments were of the so called "Zebu commercial"
type, which is mainly derived from Zebu (Bos indicus) with some genes of
European breeds (mainly Brown Swiss and some Holstein). It is the most widely
used kind of cattle for fattening in Colombia. These animals are well suited to the
tropical conditions, mainly through adaptations in their skin, which is highly
pigmented, loose and light in colour.
For Experiment 1, 24 animals were used, allocated to eight treat- ments with 3
replicates each. For the second experiment, 5 animals per treatment were used
(replicates) and the number of treatments was 8, giving a total of 40 animals.
Two animals fitted with rumen fistulae were used in Experiment 3 for the
degradability tests.
Experiment 1
Fresh sugar-cane bagasse was treated with steam in a non-continous digester
which worked under the following parameters: pressure 10 to 17 atmospheres,
temperature 180 to 200°C, treatment time 5 to 7 minutes. After this treatment a
product was obtained with a softer texture and a darker colour than the raw
bagasse. Pate (1982) and Kling et al (1987) report similar texture to that found in
this experiment.
Experiment 2
For this experiment two different treatments of the sugar-cane bagasse were used.
The first treatment was exactly the same as that already described in Experiment
1. The second treatment was a combination of physical and chemical treatments:
sugar-cane bagasse treated with steam (as described for Experiment 1) was then
treated with ammonia. This last treatment consisted of the injection of anhydrous
ammonia into a plastic (polythene) bag, which contained the steam-treated
bagasse. After treatment the bag was kept sealed for a period of 15 days before
use. The quantity of NH3 injected into the bag corresponded to 3% of the weight
of the bagasse on a dry basis. The texture of the steam-treated bagasse was not
noticeably improved by the treatment with anhydrous ammonia and, in general,
there were no apparent changes in the steam-treated bagasse after this later
chemical treatment.
Supplements
The levels of supplements fed were in proportion to the live weight of the
animals, adjusted every 28 days. Thus, where the level is described as 1% of live
weight, 1 kg fresh material was offered per 100 kg live weight.
Gliricidia sepium
For Experiment 1, this supplement was used at two different levels: 2 and 3% of
animal live weight (fresh matter basis). The percentages of Gliricidia
sepium used in the second experiment were: 1 and 1.5% of animal live weight on
a fresh matter basis.
Rice polishings
For both Experiments 1 and 2, this supplement was included in the diet at a
quantity equivalent to 0.2% of animal live weight (fresh matter basis).
Poultry litter
This was litter with a rice bran and/or wood shavings base from broiler houses.
The level of inclusion of this supplement in the diet was 0.2% of the animals' live
weight (fresh matter basis).
A restricted system (in several of the treatments in Experiment 1), and a free-
choice system (in Experiment 2) were used, in which the molasses was the
vehicle for urea and also provided trace elements. The urea/molasses liquid
mixture used in the first experiment contained (% fresh matter basis): 85
molasses, 10 urea, 3.5 mineral/vitamin pre-mixture, 1.5 salt; for the second
experiment the urea/molasses liquid mixture was varied in its urea content (12,
14 and 16% fresh matter basis), and the level of molasses was reduced (83, 81
and 79% fresh matter basis respectively), while the percentages of
mineral/vitamin pre-mixture and salt were the same as in the first experiment.
Experimental procedure
Experiment 1
For this experiment, twenty-four Zebu steers were allocated to eight treatments
(Table 1), with three animals (replicates) in each. The animals were kept in eight
pens (one pen per treatment) of approximately 3 m wide x 10 m long, in which a
third of the area was covered by a roof and the floor of this covered area was
con- creted; the rest of the area was soil surface. The three animals (replicates)
from each treatment were kept together in the same pen.
All the animals were fed with steam-treated sugar-cane bagasse ad libitum; the
differences between treatments were the level of Gliricidia sepium (1 or 1.5% of
animal live weight, fresh basis) and the quantity of urea/molasses liquid mixture
(ad libitum, 1, 1.5 or 2 kg/animal per day) fed to each animal. Additionally the
animals were supplemented with rice bran and poultry litter (each one of them at
the level of 0.2% (fresh basis) of animal live weight). The quantities of Gliricidia
sepium, rice bran, poultry litter and the restricted levels of urea/molasses liquid
mixture were calculated on the basis of the total weight of the three animals on
the same treatment, because they were not separated in the pen in which they
were kept.
The supply of the dry ingredients of the ration was made in a wooden trough in
the following way: the treated bagasse was put in first, then the mixture of the
respective quantity of poultry litter and rice bran was sprinkled on top and finally
the foliage of Gliricidia sepium was added to the former ingredients. Once all the
ingredients were in the container, they were lightly mixed by hand. The
urea/molasses liquid mixture was provided in a separate plastic trough, beside the
other containing the dry ingredients; both of them were protected from the rain
by the roof. The animals had free access to fresh drinking water at all times of the
day. Previous to the experimental period, the animals were kept for an adaptation
period of four weeks on the diet which they were going to receive and,
immediately after this adaptation period, every animal was weighed and the
experimental period began; the experimental period lasted for a total of 141 days.
Table 1: Description of the treatments in Experiment 1
Every twenty-eight days each of the animals was weighed in order to calculate
the live weight change (kg/day) between these periods of time, and this
calculated value was the main response variable used to compare treatments.
Experiment 2.
Forty Zebu steers were used in this experiment, allocated to eight treatments
(Table 2) with five animals (replicates) in each. The animals were kept in the
same pens already described for Experiment 1; in each pen five animals were
kept, all of them belonging to the same treatment.
All the animals were supplemented with rice bran and poultry litter, both at the
level of 0.2% (fresh basis) of the animal live weight. Additionally, every
treatment was supplemented with either 1 or 1.5% (fresh basis) of the live weight
with Gliricidia sepium. The quantities of these three supplements were calculated
for the five animals on the same treatment, because they were kept toge- ther, and
this total amount fed to the group; re-calculation of these quantities was made
after every weighing (four weeks). In six of the treatments the animals were fed
with steam-treated sugar- cane bagasse ad libitum, and they were supplemented
with urea/molasses liquid mixture ad libitum, containing one of three levels of
urea (12, 14 or 16% fresh basis). In the other two treatments the animals were fed
with steam-ammonia-treated sugar- cane bagasse ad libitum and they were not
supplemented with urea/molasses liquid mixture since the bagasse treated in this
way contains ammonia-N, which can be used by the rumen micro-organisms.
The diet was supplied in the same type of troughs and in the same way already
described for Experiment 1. Fresh drinking water was available to the animals at
all times.
Table 2: Description of the treatments in Experiment 2
In this experiment, the animals were given an adaptation period in the same way
as in Experiment 1, and for the same period of time (four weeks). The
experimental period lasted for 169 days; the animals with the diet of steam-
ammonia-treated bagasse were on experiment for just 113 days, and then they
were taken out of the experiment.
Weighing of every animal in the experiment was done every twenty- eight days
and the live weight change calculated to elucidate differences between
treatments.
Experiment 3
The animals used for this experiment were kept in individual pens in order to
control their diet, which consisted of steam-treated bagasse and
urea(10%)/molasses liquid mixture, both of them fed ad libitum, and rice bran
(0.2% of the live weight); one of the animals was supplemented with Gliricidia
sepium (1.5% of the live weight) for a period of twelve days, after which the
other animal was supplemented withGliricidia sepium for another twelve days.
During the last three days of every period of twelve days, degradability tests were
conducted by the nylon bag technique (Orskov et al 1980). The bags had a pore
size of 44 µm and the dimensions were 9 cm wide x 12 cm long. The samples
analysed were raw bagasse, steam-treated bagasse and steam-ammonia-treated
bagasse; cotton wool (almost pure cellulose) was used as a fourth sample to
verify if the microbial ecosystem was optimal for the digestion of a fibrous feed
resource. For the tests, five grammes of air-dried sample, previously ground
using a mill fitted with a 2.5 mm sieve, were placed in the bag, then the bags
were carefully closed and fitted firmly to a pipe and introduced into the rumen of
the animals. After 6 hours the first set of bags was taken out, washed thoroughly
and squeezed under running water from a tap for 3 minutes and then were placed
in an oven at 70 oC for 24 hours in order to obtain the dry matter content and
weighed again. The same procedure was made for the sets of samples used to
determine degradability after 24, 48 and 72 hours. The degradability was
calculated by the difference of weight of the sample before and after incubation
in the rumen.
Statistical analysis
For the statistical analysis of the data, in Experiments 1 and Experiment 2, the
linear regression of live weight change on time was calculated for each animal,
and these values were used in the analysis of variance. For Experiment 3, the
analysis of variance were done with the data measured, since the degradabilities
were analysed for each period of incubation separately.
Results
Experiment 1
The steam-treated sugar-cane bagasse was well consumed by all the animals on
every treatment (Table 3). There were no problems of urea toxicity on any of the
treatments.
There were no significant differences (P=0.92) in live weight change between the
eight treatments evaluated; despite this, the treatment which displayed the lower
live weight change (0.567 kg/d) was that with 3% of the live weight of Gliricidia
sepium and 1.5 kg/day of urea/molasses liquid mixture, while the treatment
supple- mented with 2% live weight of Gliricidia sepium and urea/molasses
liquid mixturead libitum was the one which produced the highest live weight
change (0.753 kg/d). The analysis of variance for the same variable between the
percentages of Gliricidia sepium did not show significant differences (P=0.40)
for the two levels supplied (0.743 and 0.687 kg/d for 2% and 3% of the live
weight levels respectively), nor between the four quantities of urea/molasses
liquid mixture (P=0.76) supplemented to the animals (0.739, 0.651, 0.738 and
0.731 for 1, 1.5, 2 kg and ad libitum respectively).
Experiment 2
Liveweight (kg)
- Initial 204 177 219 273 225 234 255 239
- Final 310 281 321 357 329 337 363 333
*Daily gain 0.742 0.736 0.736 0.567 0.742 0.735 0.753 0.708
SD ±.002 ±.179 ±.115 ±.081 ±.074 ±.079 ±.092 ±.113
The analysis of variance for the live weight change between the treatments with
steam-ammonia-treated bagasse and those with steam- treated bagasse showed
significant differences (P=0.00) for the treatment of bagasse (0.297 and 0.608
kg/day for steam-ammonia and steam, respectively).
The analysis of variance of the effect on live weight change of the levels of
Gliricidia (P=0.87) and the percentages of urea in the urea/molasses mixture
(P=0.40) in the animals fed on steam-treated bagasse showed no significant
differences between the two levels of Gliricidia (0.604 and 0.611 kg/day for 1%
and 1.5% of animal live weight), the three percentages of urea in the
urea/molasses mixture (0.635, 0.624 and 0.564 kg/day for 12%, 14% and 16%
urea in urea/ molasses mixture), nor for the interaction level of Gliricidia x urea
in urea/molasses mixture. There was a tendency for reduced gain when the level
of urea in the molasses liquid mixture rose. The intake of urea/molasses liquid
mixture was slightly different for each level of urea (12, 14 or 16%) used; the
animals with the mixture at 12% of urea had an intake of 1.59 kg/day (0.190 kg
of urea/day), while in those supplemented with the mixture at 14% the intake was
1.55 kg/day (0.217 kg of urea/day) and the animals with the highest percentage
of urea in the molasses (16%) had an intake of the mixture equal to 1.31 kg/day
(0.208 kg of urea/day); there was a tendency for the animals to reduce the intake
of the mixture (but not of the urea) when the percentage of urea was increased
(Table 4).
Table 4: Summary of the results of Experiment 2 (5 animals per treatment)
Liveweight (kg)
- Initial 203 292 255 238 274 218 272 268
- Final 233 324 356 356 389 315 368 369
*Daily gain 0.268 0.325 0.587 0.683 0.671 0.576 0.553 0.575
SD ±.029 ±.061 ±.078 ±.034 ±.029 ±.055 ±.064 ±.055
No. days 113 113 169 169 169 169 169 169
Table 5: Effect of supplementation with Gliricidia sepium and treatments of the sugar-cane bagasse on its
degradability in vivo (Experiment 3)
Experiment 3
Discussion
As stated in the Results section, there were no significant differences between the
three percentages of urea in the urea/mo- lasses liquid mixture. However, the
animals supplemented with the lower one (12%) reached the highest live weight
change (0.635 kg/day), while those supplemented with the higher level (16%)
obtained the lowest value for the same variable (0.564). The decision as to which
is the best level to use must be made on the basis of the cost of the additional
quantity of molasses which is consumed when the urea/molasses liquid mixture
contains just 12% of urea. Despite the fact that the intake of urea/molasses liquid
mixtures used in Experiment 2 was inversely related to the level of urea, the
quantity of urea consumed was maintained more or less at the same value. This
suggests that the intake of the mixture was regulated in accordance with the
percentage of urea in the urea/molasses liquid mixture.
The low live weight change obtained for the animals fed with steam- ammonia-
treated bagasse could be due to an increase in pH of the rumen and maybe a
change in the proportion and/or number of the protozoal population, but since
measurements were not made, these are just speculations that require further
research. It has been stated by some researchers (e.g. Ibrahim and Pearce 1983b)
that when physical and chemical treatments are combined, the time of addition of
the chemical (before, during or after physical treatment) would play an important
role in the composition of the final product; in some studies the same authors
concluded that the best time for the treatment of bagasse with ammonium
hydroxide was during steaming.
Conclusions
There are a lot of available feed resources which could be more effectively used
as animal feeds. Some of the fibrous residues such as the sugar-cane bagasse
require physical or chemical treatments to improve their utilization. More
investigations must be carried out in order to determine the potential local
resources which could be incorporated in the diets of animals. Effort must be
focussed on the best way to use them, from both a technical and economic point
of view, appropiate dietary levels and the necessary physical or chemical
treatments. The live weight change recorded for the animals fed with the steam-
treated sugar-cane bagasse was greater than that obtained in Colombia with cattle
on pastures.
The sugar-cane factories have a surplus of bagasse and in some cases the
accumulation of this by-product has become a problem. The sugar-cane bagasse
when treated, in order to improve its degradability, is a potentially good resource
to use as the basal diet in the feeding of cattle and ruminants in general. The
treatment of bagasse with steam alone results in a big increment in the
degradability of this by-product. This type of treatment is relatively easy to apply
in the sugar-cane factories since they use steam in the process of extraction of
sugar from the sugar-cane. The subsequent treatment with ammonia of the steam-
treated bagasse did not improve its degradability but resulted in lower values.
Investigation must be made to try to identify the reason for this detrimental
effect.
In Colombia there are a lot of small farmers who use sugar-cane not for the
production of sugar but for the production of panela (a brown solid block of
sugar). Their income and standard of living could be improved if alternative
methods for using sugar-cane bagasse are developed; this system must be
investigated in order to find better small-scale treatments, which may be
chemical methods, since the small farmer does not have steam available.
Supplementa- tion with Gliricidia sepium will not be a problem for them because
this legume is widely distributed throughout rural areas of the country.
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