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Research Article
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Effect of Heat-treatment on Ruminal Protein Degradability of Wolffia Meal (Wolffia globosa L. Wimm) |
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Anut Chantiratikul
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Songsak Chumpawadee
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ABSTRACT
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The objective of this research was to reduce ruminal protein degradability of Wolffia meal (Wolffia globosa L. Wimm.) by roasting at 150°C for 2 to 8 min. Two Thai-indigenous x Brahman crossbred cattle were used to determine the in situ ruminal degradability of heat-treated Wolffia meal. The ruminal degradation characteristics of Wolffia meal were determined using nylon bags incubated in rumen in reverse order for 72, 48, 24, 12, 8, 4 and 2 h. The results found the decreased rapidly soluble Crude Protein (CP) fraction (p<0.05) and the increased potentially degraded CP fraction (p<0.05) of Wolffia meal roasted at 150°C for 6 to 8 min. However, protein degradability of Wolffia meal was not reduced by heat treatment. The results indicated that roasting at 150°C for 6 to 8 min could not protect Wolffia meals protein degradation by rumen microorganism.
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Received: November 25, 2010;
Accepted: January 25, 2011;
Published: March 25, 2011
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INTRODUCTION
Wolffia meal (Wolffia spp.) one of the duckweed species, belongs to
the botanical family Lemnaceae. The family consists of five genera Landoltia,
Lemna, Spirodela, Wolffia and Wolffiella (Les et al.,
2002). Wolffia meal has been used as a vegetable in the Indochinese peninsular
for many generations (Bhanthumnavin and Mcgarry, 1971).
Interestingly, the actual dry matter yield from commercial-scale cultivation
of Lemna, Spirodela and Wolffia species in Bangladesh ranges
rom 13-38 tons/ha/year, which is a rate exceeding single-crop soybean production
six to ten fold (Skillicorn et al., 1993). Furthermore,
the protein content (29.9-45 g/100 g) of Wolffia meal grown in enriched water
containing mineral media or effluents from agricultural waste lagoons, is greatly
increased over that from natural waters low in nutrient (Skillicorn
et al., 1993; Huque et al., 1996).
Its protein content (Skillicorn et al., 1993) and
amino acid profile (Chantiratikul et al., 2010a)
were comparable to those of soybean meal. Thus, Wolffia meal can be used as
a source of protein in animal diets.
Utilization of duckweed species, namely Lemna gibba, Lemna perpusilla,
Spirodela punctata and Lemna minor as a protein source, has been
studied in ruminant animals such as cattle (Chewewattanagool,
2002) and sheep (Damry et al., 2001). However,
there is little scientific work to study on ruminal protein degradability of
duckweed. Leng et al. (1995) suggested that duckweed
would be a source of ruminally degradable protein and/or minerals to support
rumen microbial growth. Huque et al. (1996) also
revealed that duckweed proteins were highly degraded in the rumen of cattle.
On the other hands, Spirodela punctata has been found to be a valuable
source of undegradable protein for ruminants (Damry and
Nolan, 2002; Damry et al., 2001). Protein
of Wolffia meal has been firstly reported to be extensively degraded in the
rumen of bull by Huque et al. (1996). This result
indicated that only small amounts of amino acids would have been available from
Wolffia meal for absorption in the small intestine. Therefore, reduction of
Wolffia meals protein degradable in the rumen is needed to be studied.
For many years different physical and chemical methods have been researched
to reduce rumen protein degradability. Heat treatments seem to be more effective
and practical than chemical ones protecting protein from ruminal degradation
(Mustafa et al., 2000). Furthermore, heat treatments
have been reported to significantly reduce ruminal protein degradability in
many feedstuffs (Ljokjit et al., 2003; McNiven
et al., 2002). Thus, the objective of this research was to decrease
protein degradability of Wolffia meal (Wolffia globosa L. Wimm., accession
number GQ221774) by roasting at 150°C for 2 to 8 min.
MATERIALS AND METHODS
This study was conducted during October 2009 to September 2010. Two Thai-indigenous
x Brahman crossbred cattle with an average body weight of 400±30 kg were
used to determine the in situ ruminal degradability of Wolffia meal.
The animals were fitted with a rumen fistula. The experimental animals were
dewormed by Ivomectin (1 mL/30 kg BW) and injected with AD3E vitamin-complex
(1 mL/50 kg BW) before the beginning of the study. The cattle were housed in
individual pen and fed twice daily (830 and 1630 h) in equal aliquots at 1.5%
DM of body weight. The basal diet was formulated to meet nutrient requirements
of beef cattle (NRC, 1996) and prepared in total mixed
ration with rice straw as a roughage source (Table 1). Drinking
water was freely available. The experiment consisted of 14 day adjustment period
and 14 day determination of in situ ruminal degradability.
Fresh Wolffia meal (Wolffia globosa L. Wimm.) was purchased from a local
producer, who cultivated Wolffia meal as human food and dried under sunlight
for 1-2 days. Dried Wolffia meal was ground through 2 mm screen sieve. Ground
Wolffia meal was roasted in a roaster at 150°C for 2, 4, 6 and 8 min. A
portion of each sample was ground though a 1 mm screen sieve and used for analysis
of Dry Matter (DM), Organic Matter (OM), Crude Protein (CP) using the procedure
of AOAC (1990). The Neutral Detergent Fiber (NDF), Acid
Detergent Fiber (ADF) and Acid Detergent Insoluble Nitrogen (ADIN) were determined
according to Van Soest et al. (1991).
Approximately 4 g of untreated and heat-treated (at 150°C for 2, 4, 6 and
8 min) Wolffia meal was weighed and put into a nylon bag (9x15 cm) with an average
pore size 45 μm (International Feed Resources Unit, Aberdeen, UK). The
samples were prepared in four replicates. All nylon bags were soaked into the
water for 5 min to exclude the air and then introduced into the rumen of each
cannulated cattle in reverse order for 72, 48, 24, 12, 8, 4 and 2 h. After incubation,
the bags, including the 0-h bags were removed and immediately rinsed under tap
water until the water was cleared. The samples were dried at 60°C until
a constant weight was achieved. The same incubated samples within cattle were
pooled and ground through a 1 mm screen prior to analysis of DM, OM and CP according
to AOAC (1990).
Dry matter degradability was calculated as the difference between the original
sample and the weight of the residue. Nutrient degradability was calculated
as the difference between the amount nutrient (g) of the original and the residual
samples (Maiga et al., 1996). Dry matter, OM
and CP ruminal degradability characteristic were calculated according to the
monoexponential equation proposed by Orskov and McDonald
(1979).
Table 1: |
Ingredients and chemical composition of the basal diet |
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1The premix provided per kilogram of diet: 10,000
IU vitamin A; 2,000 IU vitamin D3; 20 IU vitamin E; 0.01 g Cu; 0.08 g Mn;
0.04 g Zn; 0.05 g Fe; 0.0008 g I; 0.0003 g Co; 0.0003 g Se; 0.005 g Ethoxiquin;
and 0.05 g SiO2, 2ME = Metabolisable energy |
The equation used was p= a+b (1-e-ct) where, P is the degradation
at t time (%), a is the intercept at zero time of the degradation curve (%),
b is the fraction of DM, OM and CP which will be degraded potentially (%), c
is the degradation rate constant of fraction b (/h) and t is time of incubation
(h).
The effective degradability (ED) of DM, OM and CP were estimated using the
equation of Orskov and McDonald (1979) ED = a+b {c/(c+k)}
where, a, b and c are as defined previously. k is the fractional outflow rate,
assumed to be 0.08 h-1.
Statistical analyses: Chemical compositions of the test samples were
analyzed in Completely Randomize Design using the analysis of variance (ANOVA)
procedure of SAS (SAS, 1996). Ruminal degradation data
was analyzed in Randomize Completely Block Design using the ANOVA procedure
of the SAS program. The effect of roasting time on ruminal degradation characteristics
was tested in Completely Randomize Design by the ANOVA of the SAS program. Means
were separated by Duncans New Multiple Range Test (Steel
and Torries, 1980).
RESULTS AND DISCUSSION
Wolffia meal cultivation has been recently commercialized in northeastern Thailand.
The cultivated Wolffia meal was successfully tested as protein replacement for
soybean meal in diets of poultry (Chantiratikul et al.,
2010a-c). On the other hand, there is insufficient
information of using Wolffia meal as a protein source in the diet of ruminants.
The first report that was studied on the potentiality of Wolffia meal as a feed
for cattle found that protein of Wolffia meal was mostly degraded in the rumen
after incubation for 72 h (Huque et al., 1996).
The current study tried to reduce protein degradation of Wolffia meal by roasting
at 150°C for 2 to 8 min. The results indicated that heat treatment did not
alter DM, OM, Neutral Detergent Fiber (NDF) and Acid Detergent Fiber (ADF) contents
of Wolffia meal. However, Acid Detergent Insoluble Nitrogen (ADIN) of Wolffia
meal increased significantly (p<0.05) from 7.58 to 21.08% of N when Wolffia
meal was roasted at 150°C for 2 to 8 min (Table 2).
Table 2: |
Chemical composition (% of DM) of heat-treated Wolffia meal |
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abcMeans in the same row with different superscripts
are significantly different (p<0.05), NDF = Neutral detergent fiber,
ADF = Acid detergent fiber, ADIN = Acid detergent insoluble nitrogen |
Table 3: |
Dry matter degradability characteristics and effective degradability
of heat-treated Wolffia meal |
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1a = Rapidly soluble fraction, b = Potentially
degradable fraction, c = Fractional degradation rate of fraction b, ED =
Effective degradability at an outflow rate (fraction/h) of 0.08 h-1 |
Table 4: |
Organic matter degradability characteristics and effective
degradability of heat-treated Wolffia meal |
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abcMeans in the same row with different superscripts
are significantly different (p<0.05), 1a = Rapidly soluble
fraction, b = Potentially degradable fraction, c = Fractional degradation
rate of fraction b, ED = Effective degradability at an outflow rate (fraction/h)
of 0.08 h-1 |
Table 5: |
Crude protein degradability characteristics and effective
degradability of heat-treated Wolffia meal |
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abMeans in the same row with different superscripts
are significantly different (p<0.05), 1a = Rapidly soluble
fraction, b = Potentially degradable fraction, c = Fractional degradation
rate of fraction b, ED = Effective degradability at an outflow rate (fraction/h)
of 0.08 h-1, RUP = Ruminal undegradable protein |
Similarly McNiven et al. (2002) found the increased
ADIN (3 to 9.5% of N) in soybean heated at 195°C for 3 to 6 min. The increased
ADIN in some feedstuffs also found with increased heat intensity (McNiven
et al., 2002; Pereira et al., 1988).
The present and previous results consistently demonstrated that the Maillard
reaction depends on treatment temperature and treatment time. Additionally,
moisture and reducing agents may be limiting for Maillard reaction (Martins
et al., 2001; Bach, 1997). The increase in ADIN
when heating feedstuffs is attributed to the formation of Maillard products,
which decreases rumen protein degradation (Ljokjit et
al., 2003).
The protein of untreated Wolffia meal in the present study was highly degraded
(98%) after incubated in the rumen for 72 h. Huque et
al. (1996) also found Wolffia meals protein was mostly degraded
in the rumen of bull. On the other hand, the results in sheep demonstrated that
about half of the protein in duckweed (Spirodela punctata) was degraded
in the rumen (Damry and Nolan, 2002). The different
rumen degradable protein of untreated Wolffia meal and Spirodela punctata
may be due to the incubation period in the rumen, drying conditions of duckweed
or ruminal conditions between animal species (Damry et
al., 2001).
The rapidly soluble DM fraction, the potentially degraded DM fraction, breakdown
rates (c) of the DM fraction and effective DM degradation of Wolffia meal were
not influenced (p>0.05) by heat treatment (Table 3). Heat
treatment significantly decreased (p<0.05) the rapidly soluble OM and CP
fractions, but markedly increased (p<0.05) the potentially degraded OM and
CP fractions. However, fractional degradation rate of OM and CP fractions, effective
OM and CP degradations and ruminal undegradable protein (RUP) of Wolffia meal
were not changed (p>0.05) by heat treatment (Table 4, 5).
The present results reflect that roasting at 150°C for 2 to 8 min did not
alter DM degradability, but partly affected CP degradability characteristics
of Wolffia meal. Other studies similarly observed the decreased soluble fraction
and the increased potentially degraded CP fraction of soybeans (Nowak
et al., 2005; Chouinard et al., 1997)
and whole cottonseeds (Arieli et al., 1989) by
heat treatment as a result of rapidly degradable protein fraction denatured
and became slowly degradable fractions (Van Soest, 1987).
In the current study, ADIN and potentially degraded CP fraction increased 11.61-13.50
and 8.71-9.27%, respectively. However, RUP slightly increased (2.28-2.48%) when
Wolffia meal was roasted at 150°C for 6 to 8 min. The results indicated
that roasting at 150°C for 6 to 8 min could not strongly protect protein
of Wolffia meal from microbial degradation. Haugen et
al. (2006) implied that ADIN might not be a good predictor of the indigestible
dietary protein in some forages. Additionally, ADIN can be digested in some
extent (Arieli et al., 1989). Van
Soest (1987) suggested that an optimum heat intensity and time depend on
many factors: moisture content, carbohydrate content and composition, protein
content, the presence of sulphite and therefore, optimum parameters of heat
treatment and time vary from one dietary protein to another. Therefore, further
research is needed to investigate the optimum heat intensity and duration to
reduce ruminal protein degradation of Wolffia meal.
CONCLUSION
Roasting at 150°C for 6 to 8 min decreased (p<0.05) the rapidly soluble
CP fraction, but increased (p<0.05) the potentially degraded CP fraction
of Wolffia meal when compared with those of untreated Wolffia meal. However,
heat treatment did not decrease ruminal degradation of Wolffia meals protein.
ACKNOWLEDGMENTS Mahasarakham University funded this study (Grant No. 5301106/2553) in budget fiscal year 2010. Faculty of Veterinary and Animal sciences and Mahasarakham University farm provided laboratory facilities and experimental place, respectively.
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