Effect of Burma Padauk (Plerocarpus indicus), Rain Tree (Samanea saman (Jacg.) Merr.) and Siamese Rough Bush (Streblus asper) Leaves as Fiber Sources in Total Mixed Ration on in vitro Fermentation
The objective of this study was emphasized on effect
of leaves as fiber sources in total mixed ration on in vitro fermentation
using in vitro gas production technique. The experimental was designed
in CRD with five replicates per treatment. The fiber sources in total
mixed ration were corn cob (control group), Burma padauk leaves, rain
tree leaves and Siamese rough bush leaves. The results showed that the
kinetic of gas production and digestibility were statistical significantly
differences among treatment (p<0.05). The corn cop as fiber source
in total mixed ration gave the highest potential of extent of gas production.
However, highest rate of gas production and digestibility were observed
in the Siamese rough bush leaves as fiber source. Ruminal fermentation
end-products consisted of ammonia nitrogen and volatile fatty acid were
significantly differences among treatments (p<0.05). All treatment
means were within the normal range. The pH values were relatively stable
at 7.0-7.3. The results demonstrated that Burma padauk leaves, rain tree
leaves and Siamese rough bush leaves can be used as fiber sources in total
mixed ration. Importantly, leaves are abundant and available for feeding
the ruminants in dry season.
to cite this article:
S. Chumpawadee and O. Pimpa, 2009. Effect of Burma Padauk (Plerocarpus indicus), Rain Tree (Samanea saman (Jacg.) Merr.) and Siamese Rough Bush (Streblus asper) Leaves as Fiber Sources in Total Mixed Ration on in vitro Fermentation. Asian Journal of Animal and Veterinary Advances, 4: 1-8.
In recent years, feeding a total mixed ration (TMR) for cattle has become widely
accepted. The benefits of a TMR include increased milk production, enhanced
use of low cost alternative feed ingredients, ability to control the forage
concentrate ratio, lower incidence of metabolic and digestive disorders and
reduced labor input for feeding. Fiber source of TMR is very importance, because
it can be affected feed intake, chewing activity, digestibility and production
(Chumpawadee and Pimpa, 2008). Generally, silage, forage,
rice straw corn cop and hay are conventional roughages found in TMR. Due to
the dry season have shortage fiber source for mixed TMR. Therefore, non-conventional
roughage such as fodder tree is needed for fiber source in TMR. Although, they
have the crucial parameters affecting fodder utilization, such as tannins saponin
and non protein amino acids, which are toxic to rumen microbes or to the animal
(Lowry et al., 1996). However, leaves of fodder
trees should be used as fiber sources in TMR. Because of their feed are high
content of protein, minerals and vitamins (Baloyi et al.,
1997) and availability in the dry season. In addition, the toxic substance
in leaves can be reducing by sun dry.
With respect to leave of fodder trees, limited information is available
on its use as a fiber source of TMR. The aim of this study was to investigate
the in vitro fermentation using TMR from different fiber sources.
MATERIALS AND METHODS
Preparation of TMRs
The Burma padauk(Plerocarpus indicus), rain tree (Samanea saman
(Jacg. Merr.) and Siamese rough bush (Streblus asper) leaves and
corn cop (control) were used in this study. Theywere collected from the Mahasarakham
province area in the North-East of Thailand. Fresh samples (1 kg) were hand
harvested from three site specimens. Duplicate fresh samples (0.5 kg/replicate)
were dried in a hot, dry air force oven at 65°C for 72 h and weighed. All feed
samples (Table 1) were ground to pass through a 1 mm screen
for chemical analysis. The feedstuff samples were analyzed for Dry Matter (DM),
Crude Protein (CP) and ash, neutral Detergent Fiber (NDF), Acid Detergent Fiber
(ADF) and Acid Detergent Lignin (ADL) (Van Soest et al.,
Four TMRs were formulated, to have similar total digestible nutrient
(TDN), CP, NDF, ADF, but differ in fiber source (Table 2).
The experiment was done at division of animal science, Faculty of veterinary
medicine and animal science, Mahasara Kham University, Thailand, from
August 25, 2007 to June 5, 2008. The experiment was designed in CRD with
five replicates per treatment. The fiber sources of the total mixed ration
were corn cop (control), Burma padaukleaf, rain tree leaf and Siamese
rough bush leaf. Four TMRs for the gas production test were ground to
pass through a 1 mm screen in a hammer mill.
In vitro Gas Production Test
Strict anaerobic techniques were used in all steps during the rumen fluid
transfer and incubation period. Rumen fluid inoculums was removed before the
morning feeding under vacuum pressure via the rumen fistula into a 2 L glass
flask and transferred into two pre-warmed 1 L thermos flasks which were then
transported to the laboratory. The medium preparation was as described by Makkar
et al. (1995). Mixed rumen fluid inoculums were obtained from two
fistulated Brahman-Thai native crossbred steers (weighing about 250±15 kg).
The animals were offered rice straw on ad libitum and fed 0.5% body weight
of concentrate (concentrate mixture: 49.80% cassava chip, 17.5% rice bran, 14.60%
palm meal, 7.0% soybean meal, 1.40% urea, 0.4% salt, 1.0 % mineral mix and 8.30%
sugarcane molasses). The animals were fed twice daily; water and a mineral lick
were available ad libitum for 14 days.
The feed sample of approximately 500 mg on a fresh weight basis was transferred
into a 50 mL serum bottle (Sommart et al., 2000).
The bottles were pre-warmed in a hot air oven at 39°C for about 1 h prior to
injection of 40 mL of rumen fluid medium (using a 60 mL syringe) to each bottle.
The bottles were stoppered with rubber stoppers, crimp sealed and incubated
in a hot air oven set at 39°C.
The rate of gas production was measured by reading and recording the amount
of gas volume after incubation using a 20 mL glass syringe connected to the
incubation bottle with a 23 gauge, 1.5 inch needle. Readings of gas production
were recorded from 1 to 72 h (hourly from 1-12 h, every 3 h from 13-24 h, every
6 h from 25-48 h and every 12 h from 49-72 h) after incubation periods. Amounts
of cumulative gas volume at 2, 4, 6, 12, 24, 48 and 72 after incubations were
fitted using the equation (Orskov and McDonald, 1979):
y = a+b [(1-Exp(-ct)]
|| The intercept, which ideally reflects the fermentation of the soluble
||The fermentation of the insoluble fraction
||Rate of gas production, (a+b) = potential extent of gas production
||Gas production at time t
In vitro digestibility of dry matter and organic matter was measured
at 72 h after incubation. The residues of the TMRs were removed by filtering
through a glass filtering crucible, residue was washed with 250 mL boiled
distilled water and the amount of DM and OM in the residue was estimated.
Calculation of in vitro DM and OM digestibility as a percent of
total DM and OM followed the equation:
In vitro Fermentation Measurement
The bottles were sampling at 0, 3, 6, 9 and 12 h after incubation. Rumen
fluid medium pH was measured immediately after sampling using a portable pH
meter. The rumen fluid medium was acidified with 5 mL 6 N HCl and centrifuged
at 3000 rpm for 15 min and the clear supernatant was stored in plastic tubes
at -20°C until analyzed for ammonia nitrogen (Bremner and Keeney, 1965) and
total volatile fatty acid concentration (Briggs et al.,
All data obtained from the trials were subjected to the analysis of
variance procedure of statistical analysis system according to a completely
randomized design. Means were separated by Duncan New`s Multiple Range
Test. The level of significance was determined at p<0.05.
RESULTS AND DISCUSSION
Chemical Composition of Feedstuffs and TMRs
The feed ingredients varied widely in terms of composition. The leaves
have high NDF content, more than 40.5% (Table 1). All
TMRs had a similar chemical composition. The ration CP, ash and NDF content
were approximately 11.8, 7.8 and 41.7 %, respectively (Table
Gas Production Characteristics of TMRs
A comparison of the gas production characteristics of different treatments
indicated significant differences between treatment (p<0.05). The a intercept
value for all TRMs ranged from -4.2 to -8.7 mL. The values for a, intercept,
were negative in the incubations of all TMRs in this study (Table
3). These data suggested that a lag phase due to a delay in microbial colonization
of the substrate may occur in the early state of incubation. Blummel
and Becker (1997) have also reported negative values with various substrates
when using mathematical models to fit gas production kinetics.
|| Chemical analysis of feedstuffs used for feed formulation
in the experiments (% of DM)
|DM = Dry Matter, OM = Organic Matter, CP = Crude Protein,
NDF = Neutral Detergent Fiber, ADF = Acid Detergent Fiber, ADL = Acid
|| Feed formulation and chemical composition of dietary
|*Calculated value, 1C-TMR = Corn cob as fiber
source, B-TMR = Burma padauk leaf as fiber source, R-TMR = Rain tree
leaf as fiber source, S-TMR = Siamese rough bush leaf as fiber source,
2DM = Dry matter, OM = Organic matter, CP = Crude protein,
NDF = Neutral detergent fiber, ADF = Acid detergent fiber, ADL = Acid
|| Gas production characteristics and in vitro
digestibility of dietary treatments using in vitro gas production
|a, b, c,d Means within a row different superscripts
differ (p<0.05), 1C-TMR = Corn cob as fiber source,
B-TMR = Burma Padauk leaf as fiber source, R-TMR = Rain Tree leaf
as fiber source, S-TMR = Siamese Rough Bush leaf as fiber source,
2a = the intercept (mL), which ideally reflects the fermentation
of the soluble fraction, b = The fermentation of the insoluble fraction
(asymptote) (mL), c = Rate of gas production (%/h), |a|+b = Potential
extent of gas production (mL)
This is due to either a deviation from the exponential cause of fermentation
or delays in the onset of fermentation due to the microbial colonization.
It is well known that the value for absolute |a|, described ideally, reflects
the fermentation of the soluble fraction. In this study the |a| was highest
for C-TMR and significance difference (p<0.05) with B-TMR and R-TMR.
It is indicated that the soluble fraction in C-TMR was also highest. The
soluble fraction makes it easily attachable by ruminal microorganisms
and leads to much gas production (Table 3). The soluble
fraction of corn cop was higher than leaves. It`s possibly structure and
solubility characteristics of carbohydrate and protein in corn cop which
easily attach with microorganisms in the rumen. While fodder trees had
high fibrous content it is difficult to attach by microorganisms. Generally,
leaves have a large proportion of lignified cell walls (Table
1) with low fermentation rates and digestibility. Therefore, Neutral
Detergent Fiber (NDF), Acid Detergent Fiber (ADF) and Acid Detergent Lignin
(ADL) content in fodder tree are the crucial parameters affecting fodder
The gas volume at asymptote (b) described the fermentation of the insoluble
fraction. The gas volume at asymptote was significantly higher in C-TMR than
that B-TMR, R-TMR and S-TMR (p<0.05). The gas volumes at asymptote have the
advantage of predicting feed intake. Blummel and Ørskov
(1993) found that the gas volume at asymptote could account for 88% of variance
in intake. Sommart et al. (2000) suggested that
gas volume is a good parameter from which to predict digestibility, fermentation
end-product and microbial protein synthesis of the substrate by rumen microbes
in the in vitro system. Additionally, in vitro dry matter and
organic matter digestibility were shown to have high correlation with gas volume
(Sommart et al., 2000). In this study, C-TMR showed
the highest gas volume (Table 3). Low gas volumes at asymptote
were observed in B-TMR and R-TMR. It`s possibly reflected by phenolic compounds,
which are toxic to rumen microbes (D`Mello, 1992). Getachew
et al. (2000) reported that the concentration of phenolic compounds
(particularly tannin) in tree leaves is generally high. Lowry
et al. (1996) have also reported that many of the multi propose tree
are problematic as feed supplements because they often contain anti-nutritional
compounds, such as tannin, saponin and non-protein amino acid, which are either
toxic to rumen microbe or to the animal, or their metabolic products are toxic.
Rate of gas production (c) expressed in %/h as ranked from the fastest
to the slowest were; B-TMR, S-TMR, C-TMR and R-TMR. Fast rates of gas
production were observed in B-TMR and S-TMR. This result might have been
influenced by the carbohydrate fraction that was readily available to
the microbial population.
The potential extent of gas production (|a|+b) of C-TMR was the highest and
significantly different (p<0.05) with B-TMR, R-TMR and S-TMR. This implies that
C-TMR was highly fermentable in the rumen. Remarkably, the potential of gas
production for B-TMR and R-TMR was slightly lower compared with C-TMR and S-TMR,
possibly due to the influence of carbohydrate fraction in the TMR. Fibrous constituents
had negatively influenced in vitro gas production (Melaku
et al., 2003). The fiber content of the B-TMR and R-TMR were high,
when compared with C-TMR and S-TMR. The high fibrous content is difficult to
attach by microorganisms. Therefore, less gas production was shown in the B-TMR
In vitro Digestibility of Dry Matter and Organic Matter
It can be seen that IVDMD and IVOMD are similar. The IVDMD and IVODM were
significantly different (p<0.05) among treatment. The S-TMR gave the highest
IVDMD and IVOMD. This result implies that the microbe in the rumen and animal
have high nutrient uptake (Table 3). The IVDMD and IVOMD of
C-TMR, B-TMR and R-TMR lower than S-TMR. The reason for that is possibly that
the fiber fractions of C-TMR, B-TMR and R-TMR have a large proportion of lignified
cell walls leading to attachment difficulty by microorganism, with low fermentation
rates, low digestibility rate and limited intake (Ibrahim
et al., 1995). The higher fiber content (Table 1)
of Burma Padauk leaf and rain tree leaf probably resulted in lower in vitro
dry matter and organic matter digestibility since high NDF and ADL content in
feedstuffs result in lower fiber degradation (Van Soest,
In vitro Fermentation Pattern
Concentrations of NH3-N, TVFA and pH in the in vitro fluid
were used to monitor the in vitro fermentation pattern (Table
4). The pH was not affected by fiber source in TMRs. When monitoring pH
pattern at 0, 3, 6, 9 and 12 h after incubation, the pH values were relatively
stable at 7.0-7.3 and all treatment means were within the normal range that
has been reported as optimal pH (6.0-7.0) for microbial digestion. The buffer
in the rumen fluid medium is the reason for pH remaining stable at all times
of fermentation. The buffer is a factor that should be considered when using
the gas production technique. The exhaustion of the buffer would lead to a lowering
of the pH (Getachew et al., 1998).
|| Effect of Burma Padauk (Plerocarpus Indicus),
Rain Tree (Samanea Saman (Jacg.) Merr.) and Siamese Rough Bush
(Streblus Asper) Leaves as Fiber Sources in Total Mixed Ration
on pH, ammonia nitrogen (NH3N) and total volatile fatty
acid (TVFA) in in vitro
|a, b, c, dMeans within a row different superscripts
differ (p<0.05), 1C-TMR = Corn cob as fiber source,
B-TMR = Burma Padauk leaf as fiber source, R-TMR = Rain tree leaf
as fiber source, S-TMR = Siamese rough bush leaf as fiber source
At a lower pH, the celluolytic bacteria becomes less active (Russell
and Dombrowski, 1980). In this study the buffer was not exhausted. Therefore,
this condition was optimal for microbial activity.
Ammonia nitrogen concentration was significantly different (p<0.05) among treatments
at each hour of sampling, excepted at 0 and 12 h of sampling. The difference
in NH3-N concentrations among treatments may have been related directly
to urea and degradability of protein in the TMRs. Although, nitrogen recycling
in the rumen and in vitro is different, NH3-N concentration
was in the optimal range for rumen ecology, microbial activity (Perdok
and Leng, 1990; Wanapat and Pimpa, 1999). At 3 to
9 h after incubation C-TMR had the highest NH3-N, when compared with
other TMRs. When ammonium nitrogen is high it indicates that the soluble fraction
of protein is also high. Remarkably, NH3-N concentration of B-TMR
and R-TMR were low at all time of sampling. It may have been that the urea level
in both TMRs was lower than C-TMR. In addition, the protein in Burma Padauk
leaf and rain tree leaf had low degradability (Chumpawadee
et al., 2007). Although, NH3-N concentration of all TMRs
was different with C-TMR (control), it was in the normal range. Therefore, it
can be used as fiber sources in the TMRs. Future research should investigate
the impact of the ability of leaves feed to replace forage in intact animals.
Total volatile fatty acid concentrations were significantly different (p<0.05)
among treatments at all times of sampling, excepting 0 h after incubation. Remarkably,
TVFA concentrations in the in vitro medium, from 0 to 12 h after incubation,
tend to be increased. The reason for that is possibly VFAs accumulated in the
medium. The VFA can not absorb via the in vitro, but most of the VFA
can be absorbed into rumen wall. Although, VFA increased in the medium, pH did
not change because the buffer in the medium was not exhausted. This is the advantages
of the gas production technique. The VFA production of B-TMR are the same with
S-TMR and difference from C-TMR and R-TMR, this result might have been influenced
by carbohydrate fraction in TMRs. The rate and extent of carbohydrates degradation
are influenced by the condition of rumen fermentation and rate and extent of
VFAs production (Cheng et al., 1991). Keady
and Mayne (2001) also suggest that VFAs concentration is similar when the
animal fed diets contained a similar carbohydrate composition. In this study,
have difference source of fiber in TMRs, thus VFA concentration was also different.
In in vitro study, leaves as fiber source in TMRs had an affect
on gas production characteristic, in vitro digestibility and in
vitro fermentation. The corn cop as a fiber source in TMR gave the
highest parameters of gas production characteristic. The Siamese rough
bush as fiber source in TMR gave the highest in vitro digestibility.
Concentration of NH3-N, TVFA and pH were different when TMRs
contained a different fiber sources, but they are in the normal range.
Therefore, leaves can be used as fiber source in TMRs. Future research
should investigate the impact of a leaves feed replacement of forage for
The authors would like to express their gratitude to all staff and my
student for their invaluable help on the farm and laboratory. We are grateful
to the Division of Animal Science, Faculty of Veterinary Medicine and
Animal Science, Mahasarakham University for supporting experiment facilities.
This research project No. MRG5080362 was funded by Thailand Research Fund
and Commission on Higher Education under the program New Researcher Grant.
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