Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online)
An Open Access, Online International Journal Available at http://www.cibtech.org/jls.htm
2014 Vol. 4 (2) April-June, pp. 396-401/Shadmanesh
Research Article
© Copyright 2014 | Centre for Info Bio Technology (CIBTech) 396
EFFECT OF DIETARY SUPPPLEMENT WITH FIBROLYTIC ENZYMES
ON THE PRODUCTIVE PERFORMANCE OF EARLY LACTATING
DAIRY COWS
*Ali Shadmanesh
Faculty of Veterinary Medicine Islamic Azad University – Eghlid Branch, Iran
*Author for Correspondence
ABSTRACT
An experiment was conducted to study the effects of supplementing the diets of early lactating dairy cows
with exogenous fibrolytic enzymes (Fibrozyme, Alltechinc Company, USA) on feed intake, feed
efficiency, milk production and composition of milk for 70 days. 28 Holstein dairy cows at early lactation
(50 ± 6.2 days in milk) were randomly divided into two groups according to lactation period "stage of
lactation" and lactation season. The first group (control, n=14) were fed total mixed ration (TMR) without
a supplement of fibrolytic enzymes. The second group (treatment, n=14) were fed TMR supplemented
with a commercial fibrolytic enzymes at the rate of 15 g/cow/d for 70 days. Each group was placed in an
open pen. Fibrolytic enzyme was added to TMR at the time of feeding twice per day at 8 am and 15 pm.
The results showed that treatments had no effect on dry matter intake. Milk yield (4%) and the fat
corrected milk were increased as a response to fibrolytic enzymes supplementation to lactating dairy cows
compared to un-supplemented dairy cows (P<0.005). In addition, feed efficiency in treated was increased
significantly than of the control group (P<0.001).
The milk fat, protein, lactose and solid not fat (SNF) percentages in treated were not higher than of the
control group. While, quantities of milk protein, lactose and SNF in treated were improved significantly
compared to control group except quantity of milk fat (P<0.008). It is concluded that fibrolytic enzymes
supplementation to early lactating dairy cows improved significantly milk production and SNF.
Keywords: Fibrolytic Enzymes, Productive Traits, Dairy Cow
INTRODUCTION
The digestion of plant cell wall in the rumen is slow and incomplete and can limit milk production and
increase the cost of production. Over the years, there has been a continual search for new additives that
will enhance feed utilization so that the greater nutrient demands of ever-increasing animal productivity
can be met. The availability of new enzyme products, combined with the high cost of livestock
production, has prompted researchers to examine the potential role of exogenous enzymes in ruminant
production (Beauchemin, 1995; Chen, 1995).
The use of exogenous fibrolytic enzymes (EFE) to enhance quality and digestibility of fibrous forage is
on the some of delivering practical benefits to ruminant production systems. In this matter, cellulase and
xylanase are two major enzyme groups that are specified to break ß1-4 linkages joining sugar molecules
of cellulose and xylan found in plant cell wall components (Beauchemin, 1999). Several studies with EFE
have made mention of the increase of microbial activities in the rumen, which resulted in an enhancement
of animal performance traits. Despite the increase in feed digestibility and subsequent production traits,
the relationship between the improvement in forage utilization and enzymatic activities is yet to be
explained in ruminant systems. In addition, results with EFE addition in ruminant systems are variable
and somewhat inconsistent (Beauchemin, 1999; Colombatta, 2003), making their biological response
difficult to predict.
Some studies have shown substantial improvement of feed digestibility and animal performance traits
(Arriola, 2011; Bala, 2009), while others reported either negative effects or none at all (Baloyi, 2008).
If the potential intake and/or the density of available nutrients of forages can be increased with fibrolytic
enzyme as feed additives, then poor quality forages can be economically and successfully converted into
meat and milk for human consumption. Moreover, an increase in the input costs in the dairy industry has
Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online)
An Open Access, Online International Journal Available at http://www.cibtech.org/jls.htm
2014 Vol. 4 (2) April-June, pp. 396-401/Shadmanesh
Research Article
© Copyright 2014 | Centre for Info Bio Technology (CIBTech) 397
demonstrated the need for methods to increase production efficiency. One way of increasing efficiency
would be to increase the bioavailability of nutrients in a feedstuff, which might be accomplished through
the e fibrolytic enzymes supplement. Therefore, the objective of the undertaken experiment was to
evaluate the effect of exogenous fibrolytic enzymes "Fibrozyme" supplementation on dry matter intake,
feed efficiency, milk production and milk composition in early lactating dairy cows.
MATERIALS AND METHODS
Animals’ Management and Experimental Design
The experiment was conducted at Islamic Azad University Eghlid- Branch, from August to November,
2013 for70days.28 Holstein dairy cows at early lactation (50 ± 6.2 days in milk) were randomly divided
into two treatment groups. One group (N=14) was fed TMR supplemented with enzyme mixture while the
other group (N=14) saved as control. The enzyme preparation Fibrozyme TM, used in the present study is
a blend of active xylanase and cellulase, which are a dried mixture of fermentation extracts from
Aspergillus niger and Trichoderma longibrachiatum fungi, and having xylanase activity by minimum 100
XU/g, Alltechinc Company, USA) at the rate of 15 g/cow/d according to the guide of the manufacture for
90days. Fibrozyme was added and mixed to the TMR at the time of feeding. Cows in the two groups fed a
total mixed ration (TMR, Table 1), which composed of alfalfa hay, corn silage, soybean meal, Barley
grain, Blood meal, Corn gluten meal, vitamins- minerals mixture, calcium bicarbonate, dicalcium
phosphate and mono sodium phosphate. The proximate analysis and calculated nutritive value of the
TMR is given in Table 2.
Table 1: Ingredients of the experimental total mixed ration (TMR) in the lactation trial
Ingredients % of DM
Alfalfa hay
Corn silage
Barley grain
Blood meal
Corn gluten meal
Soybean meal, 48% CP
Calcium carbonate
Dicalcium phosphate
Mono-sodium phosphate
Mineral-vitamin mix
Cr2o3
15
24
44
0.95
5.2
4
.95
0.48
0.1
1.5
0.3
Table 2: Proximate analysis (%) and calculated nutritive value of the experimental total mixed
ration (TMR) in lactation trial
Nutrients As dry matter basis (%)
DM
OM
Crude Protein
Neutral detergent fiber
Acid detergent fiber
Ash
Non Fiber carbohydrates
Ether Extract
ME (M cal/kg DM)
NEL (M cal/kg DM)
60.5
92.6
16.7
31
17.8
5.8
36.5
4.6
3.3
1.75
Cows were fed as a group open feed, with free access to water. Amount of TMR delivered was measured
with electronic scales on mixer–feeder wagon. The TMR was mixed and fed using Delaval mixer wagon.
Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online)
An Open Access, Online International Journal Available at http://www.cibtech.org/jls.htm
2014 Vol. 4 (2) April-June, pp. 396-401/Shadmanesh
Research Article
© Copyright 2014 | Centre for Info Bio Technology (CIBTech) 398
The diet was formulated using of recommendations NRC (2001), (Table2). Cows were milked twice
times daily at 5 am and 5 pm in a Dobell 10-parallel milking parlor, milk recording system, and
automated detacher milker units.
Sampling Analysis
During the experiment, samples of TMR were collected weekly and stored at -180C until chemical
analysis for dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), crude fiber
(CF), neutral detergent fibers (NDF) and acid detergent fiber (ADF). Analysis chemical of samples were
determined according to Goering and Van Soest (1970), Van Soest (1991) and AOAC, (2006).
Metabolizable and net energy for lactation were calculated according to NRC (2001).Milk samples were
collected weekly and analyzed immediately for fat, protein, lactose and SNF content using infrared
method by Milk Analyzer. Average fat and CP yields were calculated by multiplying milk yield by fat
and CP content of milk on an individual cow basis.
Statistical Analysis
Data using of software SAS (2002), version9), in a completely randomized design with a 2 factorial
(enzyme and week) arrangement were analyzed. A model containing treatment, week (repeated measure),
and all interactions.
RESULTS AND DISCUSSION
The results of the proximal analysis of TMR are shown in table 2. The results showed that the organic
matter (OM), crude protein (CP), ether extract (EE), NDF, ADF and non- fiber carbohydrate were 92.6,
16.7, 4.6, 31, 17.8 and 36.5 respectively. Metabolizable energy (ME) and net energy for lactation (NEL) in
TMR were calculated 3.3 M cal/kg DM and 1.75 M cal/kg DM respectively.
Effect of enzyme supplement of the diet of lactating dairy cow at early lactation on dry matter intake,
milk production, fat corrected milk and feed efficiency are shown in table 3.Intake of DM was not
affected by febrolylic enzymes supplementation. This is in agreement with our previous studies (Ahn,
2003; Arriola, 2011; Dyaa, 2013; Lewis, 1999; Rode, 1999; Yang, 1999), in which a high xylanase, high
cellulase enzyme product was added to either the TMR, forage, or concentrate. On the other hand, several
researchers recorded an increase in DMI of dairy cows when fibrolytic enzymes was applied to forage
before mixing with other ingredients (Lewis, 1999) or applied to TMR or concentrate portion of the diet
(Ware, 2005). However, the effects of fibrolytic enzymes on DMI appear to be vary among enzymes
products and the method of applying of enzymes (Bowman et al., 2002).
Table 3: Effect of fibrolytic enzymes supplementation on dry matter intake, milk production, fat
corrected milk and feed efficiency at lactating dairy cows
Iems Control E-TMR P values
W EF W*FE
DMI
Milk yield(kg/d)
FCM(Kg/d)
milk: feed ratio
19.5±0.15
31.2±0.3b
29.04±0.22b
1.48±0.01b
19.60.15
32.5±0.3a
30.05±0.22a
1.66±0.01a
0.875
0.017
0.017
0.001
0.875
0.003
0.025
0.001
0.955
0.943
0.950
0.937
*Different letters (a, b) in same row indicate significant differences.
*W: weeks and FE: Fibrolytic enzyme
*Fat corrected milk (FCM) = milk yield*0.4+fat yield*15
Milk yield (p</003), fat corrected milk (p<0.025) and feed efficiency (p<0.001) was significant higher for
cows fed the E-TMR diet than for cows fed the control.
In some studies, the response to enzymes has been substantial. For example, Lewis (1999) treated for age
with a cellulase/xylanase mixture (FinnFeedsInt.; supplying 1 mL/kg of total mixed ration [TMR], DM
basis) and observed that cows in early lactation produced 6.3 kg/d (16%) more milk. However, higher and
lower levels of the same enzyme product were less effective. Rode (1999) applied an enzyme product
Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online)
An Open Access, Online International Journal Available at http://www.cibtech.org/jls.htm
2014 Vol. 4 (2) April-June, pp. 396-401/Shadmanesh
Research Article
© Copyright 2014 | Centre for Info Bio Technology (CIBTech) 399
(Promote, Biovance Technologies Inc., Omaha, NE) to the concentrate portion of a diet (supplying1.3
g/kg of TMR on a DM basis) and observed a 3.6 kg/d (10%; P = 0.11) increase in milk production for
cows in early lactation. Yang (2000) added an enzyme mixture (BiovanceTechnol, Omaha, NE) to the
concentrate, and cows in early lactation produced 2 kg/d (5.9%) more milk. However, there was no
response when the same enzyme was added to the TMR. But no milk response was reported in others
(Bernard, 2010; Elwakeel, 2007). Furthermore, these enhancement in milk yield at the current study are in
line with those found by Guerra et al., (2007) who used Fibrozyme in diet containing alfalfa hay and they
reported that Fibrozyme supplementation increased milk yield, which may be due to improved utilization
of nutrients in digestive tract and in rumen and increased gain of net energy.
The improvement in feed efficiency observed in the current lactation study might be attributable to
greater NDF digestibility in the rumen and the similar trend was concluded by Holtshausen (2011).
Improvements in feed conversion efficiency were due to lower DMI rather than a change in milk yield.
Improved feed efficiency indicates better utilization of nutrients when TMR was treated with enzymes,
with the magnitude of improvement being a linear function of enzymes dosage.
The significance contrasts in Table 3 showed that there were no interaction between treatment and time,
while time had significant effects on milk yield, FCM and feed efficiency in treated group compared to
the control group.
Effect of enzymes supplement to the diet of lactating dairy cow at early lactation on milk composition are
shown in table 4. The results denoted that fibrolytic enzymes had no significant effect on milk fat, protein
lactose and solid not fat (SNF) percentage compared to the control group of dairy cows. The significance
contrasts in Table 4 showed that there were no interaction between treatment and time, while time had
significant effects on yield of milk fat, protein, lactose and SNF in treated group compared to the control
group.
Table 4: Milk composition of lactating dairy cows at early lactation fed TMR with or without
fibrolytic enzymes (FE)
Items control E-TMR P values
W EF W*FE
Fat, %
Fat, kg
Protein, %
Protein, kg
Lactose, %
Lactose, kg
NSF, %
NSF, kg
3.54±0.15
1.104±0.02
3.15±0.02
0.98±0.01b
4.7±0.15
1.47±0.014b
8.4±0.07
2.62±0.03b
3.5 ±0.15
1.137±0.02
3.25±0.18
1.06±0.01b
4.8 ±0.16
1.56±0.014a
8.6±0.07
2.79±0.03a
0.485
0.016
0.007
0.018
0.88
0.017
0.12
0.017
0.482
0.1
0. 25
0.008
0.787
0.008
0.896
0.005
0.922
0.954
0.80
0.937
0.982
0.924
0732
0.958
*Different letters (a, b) in the same row indicate significant differences.
*SE: standard error; SNF: solid not fat; W: weeks.
In agreement with our finding, several studies has been reported that fibrolytic enzymes supplementation
to Holstein dairy cows did not affect (P>0.05) on milk composition (Bernard, 2010; Dyaa, 2013; Granzin,
2005; Lewis, 1999). On the other hand, Yang (1999) and Mansour (2009) found that milk fat increased
when adding fibrolytic enzymes. The increase in fat percentage may be due to the increase in available
energy and fatty acids for fat synthesis. Gado (2009) concluded that milk protein yield for Brown Swiss
cows was (P<0.05) increased (0.57 %) for cows fed ZADO® supplemented diet compared with 0.45
kg/h/day for cows fed control diet. The variability in responses among studies may be attributed to variety
of enzyme products and experimental conditions.
Conclusion
Applying enzymes to the TMR before feeding dairy cows in early lactation improved milk production and
SNF.
Indian Journal of Fundamental and Applied Life Sciences ISSN: 2231-6345 (Online)
An Open Access, Online International Journal Available at http://www.cibtech.org/jls.htm
2014 Vol. 4 (2) April-June, pp. 396-401/Shadmanesh
Research Article
© Copyright 2014 | Centre for Info Bio Technology (CIBTech) 400
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