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Pakistan Journal of Nutrition 6 (2): 149-154, 2007
ISSN 1680-5194
© Asian Network for Scientific Information, 2007
149
Effects of Organic Acid and Bacterial Direct-Feed Microbial on Fattening
Performance of Kivircik-Male Yearling Lambs
Ismail Abas , Halil Can Kutay , Recep Kahraman , Nezir Yasar Toker ,
1 1 1 2
Dervis Özçelik , Fatma Ates and Aysun Kaçakci
3 3 3
Department of Animal Nutrition and Nutritional Diseases,
1
Faculty of Veterinary Medicine, Istanbul University, 34320 - Avcilar, Istanbul, Turkey
Department of Biochemistry, Faculty of Veterinary Medicine,
2
Istanbul University, 34320, Avcilar, Istanbul, Turkey
Department of Biophysics, Cerrahpasa Medical Faculty,
3
Istanbul University, 34300 Cerrahpasa, Istanbul, Turkey
Abstract: This study was undertaken to investigate the effects of addition of organic acid (OA), (SAL CURB®)
and/or direct-feed microbial (DFM), (Cylactin® LBC ME 10) to the rations of Kivircik male-yearling lambs on
body weight gain, feed consumption, feed conversion, some blood metabolites, and the levels of serum
minerals. A group of sixty 1-year old male lambs of Kivircik Breed were used. The animals were divided in
to 4 groups of 15 lambs each. The groups were assigned as control, organic acid, direct-feed microbial, and
direct-feed microbial plus organic acid. A commercial direct-feed microbial product and organic acid mix were
used as feed treatment supplements. The average daily gain means of the lambs supplemented with
organic acid plus direct-feed microbials was found significantly higher than that of control group lambs.
Significant differences in total proteins, cholesterol, total lipids, and serum Zn levels were detected among
the groups. As a result, addition of organic acid and/or direct-feed microbial to the ration of yearling Kivircik
male lambs resulted in an increased body weight gain (p<0.05). In addition, the dietary supplement of
organic acid to the lambs caused a decrease in the serum cholesterol level while direct-feed microbial
addition caused an increase in the serum Zn level (p<0.05).
Key words: Kivircik male lambs, organic acid, direct-feed microbial, performance, blood parameter
Introduction
Recent studies on ruminant nutrition have mostly
focused on increasing the feed conversion rate. A variety
of feed additives have been developed to achieve this
objective. Antimicrobial feed additives, which are one of
the additives that have been used widely in the world;
have been prohibited or restricted in most countries due
to the increased concern of causing resistance to
antibiotics in bacterial pathogens. Therefore, the interest
to potential alternatives to antibiotic feed additives such
as direct-feed microbials (DFM), which is known as
biological products composed of cultures of useful
microorganisms (Alp and Kahraman, 1996; Krehbiel et
al., 2003; Elam et al., 2003), and organic acids
increased (Callaway and Martin, 1996; Martin, 1998;
Castillo et al., 2004).
In ruminants, direct-feed microbials are mainly used for
lowering the effects of stress conditions and decreasing
the use of antibiotics in calves. These microbials both
increase milk yield in dairy cows and the body weight
gain and feed conversion rate in beef cattle (Krehbiel et
al., 2003). Lema et al. (2001) it has been reported that
the addition of direct-feed microbials to the ration of
sheep resulted in decreased numbers of harmful
microorganisms in the intestines improved fattening
performance, and feed conversion rate.
Few studies have been carried out on the action of DFM
on blood parameters and published results have been
conflicted. Jouany et al. (1998), reported that DFM did not
have any effect on ruminal metabolism such as protein
decomposition or microbial protein synthesis and did
not cause any changes in some of the blood parameters
including serum urea, total protein and albumin. In
contrast, Chiofalo et al. (2004), found an increase in
serum urea concentration, whereas no change was
observed in blood total protein and total cholesterol.
There is still no product or method developed as a
substitute of antimicrobial feed additives. Yet, organic
acids (OA) are thought to be as one of the most
important candidate as an alternative of antibiotic feed
additives.
Organic acids establish antimicrobial effect in the
intestines by suppressing fungal activity and maintaining
acidic environment (Dibner and Buttin, 2002). Organic
acids are reported to improve rumen fermentation, like
ionophor antibiotics, they do and maintain the rumen pH
even after consumption of carbohydrate-rich feeds
through which an increased growth and fattening
performance are observed (Martin, 1998). Martin et al.,
Abas et al.: Organic Acid and Bacterial Direct-Feed Microbial
150
Table 1: Concentrate composition of the experimental ration.
Ingredient g/kg, as mixed
Corn grain, ground 360.0
Barley grain, ground 200.0
Sunflower Meal (280 g/kg CP) 200.0
Corn bran 80.0
Wheat bran 70.0
Molasses 65.0
Limestone 18.0
Salt 5.0
Vitamin + mineral premix* 2.0
*Composition of vitamin - mineral premix per kilogram of premix:
vitamin A 12 000 000 IU; vitamin D 3 000 000 IU; vitamin E 30
3
g; Mn 50 g; Fe 50 g; Zn 50 g; Cu 10 g; I 0.85 g; Co 0.15 g; Se
0.15 g.
Table 2: Nutrient contents of diet (DM basis)
Chemical composition (g/kg) Concentrate* Forage**
Dry Matter 872.4 834.6
Ash 61.0 96.4
Crude protein (N X 6.25) 148.4 104.2
Crude fiber 96.1 337.0
Ether extract 22.3 15.6
Ca 8.1 6.9
P4.7 1.5
*Ration consisted of 1: 1 concentrate: forage. **Composition of
per kilogram of forage: grass hay 520 g, wheat straw 440 g and
corn silage 40 g, as fed.
1999, noted that plasma component concentrations
were generally unaffected by organic acid (DL-malate)
except that, plasma urea level was lower for steers
whereas cholesterol was higher for heifers. Results of
a recent study by Castillo et al. (2004) showed that, in
addition to buffering effect in feed and rumen, organic
acids might increase energy-efficiency and digestibility
of crude protein, calcium, and phosphorus by lowering
methane production and decrease the numbers of
harmful bacteria attached to the intestinal wall.
The organic acid and DFM are feed supplements to
ruminant with the claim of improving performance.
However, there is little information about OA and DFM or
their combination on performance of lambs. To further
investigate the efficacy of using this additives with along
or their combination for lambs, we conducted
experiment to compare the effects of OA and DFM or
their combination on performance and blood
parameters of Kivircik male-yearling lambs. Blood
metabolite concentrations were also evaluated for
nutritional status of lambs.
Materials and Methods
Sixty yearling Kivircik male lambs were used in the
present study. The animals were randomly divided into
four boxes of four different treatments. The four different
treatments were assigned as control (C, no additive),
organic acid (OA), direct-feed microbial (DFM), and
organic acid and direct-feed microbial combination
(OA+DFM). The direct-feed microbial used was a
commercial product of Enterococcus faecium cernelle 68
strain (Cylactin® LBC ME 10) at 1.0x10 cfu/g
10
concentration. The organic acid used was a commercial
mixture (SAL CURB® brand dry) of 1 g/kg (propionic
acid, calcium propionate, ammonium formate, sorbic
acid, formic acid, and BHA (butylated hydroxyanisole).
The concentrate of control group of animals did not
receive any additive. The animals in OA group received
a mixture of organic acids at 3 g/kg, group DFM animals
received direct-feed microbial at 1 g/kg, and OA+DFM
group of animals received a combination of organic acid
(1.5 g/kg) and direct-feed microbial (0.5 g/kg) in their
concentrates.
The treatment ration consisted of 1:1 forage: concentrate
and was fed as total mixed ration (Table 1 and 2). Fresh
and clean drinking water and feed (mixture of roughage
and concentrate-yearling lamb feed) were given to the
animals ad libitum.
The roughage and concentrate feed were analyzed for
dry matter, ether extract, Nitrogen, and crude ash using
standard procedures AOAC (1990). Dry matter was
determined by oven drying for 24 h at 105 C (DM;
o
934.01), Nitrogen using the Kjeldahl Procedure with a
Kjeltec- UDK 126 A (Velp Scientific, Italy) (CP; 954.01),
crude ash by combustion at 550 C for 6h (OM; 942.01)
o
and ether extract by using Soxhlet extraction procedure.
Each animal was ear-tagged and weighed on days 0,
28, and 56 using a scale. Feed intake was recorded
daily as the difference between what was offered and
what was refused. In addition, the feed conversion ratio
of every group was calculated. Health conditions of the
lambs were closely monitored during the study.
Blood samples were collected from the individual
animals after morning feedings on days 0, 28, and 56.
The blood samples were taken from V. jugularis in to 10
ml-vacutainer tubes without anticoagulant and
centrifuged (4000rpm/min) for the separation of serum.
Using ready-to-use spectrophotometrical kits, the serum
samples were analyzed for deterring the levels of
albumin (Chema Diagnostica BC 0500 CH), total protein
(Chema Diagnostica TP 0500 CH), urea (Chema
Diagnostica A2 F245 CH), cholesterol (Chema
Diagnostica CT F400), total lipids (Spinreact, 1001270),
Ca (Spinreact, 1001060), Mg (Chema Diagnostica TP
0500 CH), and Pi (Spinreact, 1001155). In addition,
serum Fe, Cu and Zn levels were determined using
atomic absorption spectrophotometer (AAS, Shimadzu
680 AA) according to the methods of AOAC (1990).
The data was analyzed for One-way variance analysis
(ANOVA) using SPSS statistical analysis software
(SPSS, 2002). Tukey’s HSD test was used for comparing
the means of the treatment groups. A significant level of
0.05 was used for comparisons (Snedecor and
Cochran, 1980). Statistical analysis were not conducted
for feed consumption and feed conversion ratios since
the animals were fed in-groups.
Abas et al.: Organic Acid and Bacterial Direct-Feed Microbial
151
Table 3: Body weight and average daily weight gain of yearling lambs
Groups
------------------------------------------------------------------------------------------------------
COA DFM OA+DFM SEM
Body weight, kg
Initial 47.90 46.03 48.03 44.75 1.010
Day 28 51.97 51.52 52.21 51.31 1.090
Day 56 59.90 59.75 61.49 59.52 1.134
Average daily weight gain, g
Day 0 - 28 145.1 195.9 149.1 234.4 10.40**
bab b a
Day 29 - 56 283.2 294.2 331.5 293.2 9.11*
bab a b
Day 0 - 56 214.3 244.9 240.4 263.9 7.33*
bab ab a
Groups: Control (C); Organic Acid (OA); Direct-fed microbials (DFM); Organic acid + Direct-fed microbials (OA+DFM)
Means within sub rows with no common superscripts differ significantly (p<0.05). * p<0.05; **p<0.01
a-b)
Results
The effect of dietary organic acid supplement and direct-
feed microbial on body weight (BW) and average daily
gain (ADG) of Kivircik Breed yearling male lambs are
shown in Table 3. Significant differences in ADG
between periods of days 0-28, 29-56, and 0-56 were
detected among the groups. However, during the trial,
the animals of OA, P, or P+OA groups showed no
significant change regarding body weight at day 56.
Overall ADG of the lambs in OA+P group were found
significantly higher than that of control group the trough
d 56.
Feed consumption for different periods (days 0-28, 29-
56 and 0-56) for total, roughage, and concentrate and
calculated amount of feed for 1 kg of BW gain is provided
in Table 4. Statistical analysis were not conducted for
feed consumption and feed conversion ratios since the
animals were fed in groups.
The data of the effect on lipid, protein and mineral
metabolism created by adding organic acid and/or
direct-feed microbial to the rations of Kivircik yearling
lambs are presented in Table 5. Significant differences
were found among groups at the levels of urea,
cholesterol, Mg, and Zn on day 28 and at the levels of
total proteins, cholesterol, total lipids and Zn on day 56.
Discussion
In this study, it has been investigated how the addition of
organic acid and/or direct-feed microbial to the rations of
Kivircik Breed male yearling lambs affect the feeding
performance, some blood parameters. The feed
additives are used in animal nutrition either by direct
addition to feed or water or as silage additive (Alp and
Kahraman 1996; Kurtoglu et al., 2001). In recent years,
a variety of in vitro rumen studies have been undertaken
to investigate the effects of OA and DFM additives on
rumen metabolism (Newbold et al., 1995; Callaway and
Martin, 1996; Lopez et al., 1999; Agarwal et al., 2000).
Results of the present study indicated that there were
appreciable but not significant differences among
groups in mean BW of the animals determined at the
end of the feeding period. However, the ADG of the
animals, which received OA, DFM, or OA+DFM were
significantly higher regarding in their rations than daily
gains of the control animals during the entire study
period (Table 3). For instance, BW gain of the animals in
OA+DFM group per day was 23.15% higher than that of
control group lambs (p<0.05). Although the daily feed
consumption of the animals in all groups were close to
each other, feed conversion ratios determined in
animals that received feed additive in their rations were
higher than those determined in control lambs (Table 4).
The results of the present study are consistent with other
published reports indicating that addition of direct-feed
microbial as microbial feed additive to the ration of
sheep resulted in increased BW gain and feed
conversion rate (Lubbadeh et al., 1999; Lema et al.,
2001; Chiofalo et al., 2004). Henderson et al. (1986)
reported that feeding a sheep with silage inoculated
through bacterial cultures resulted in an increased
consumption of dry substances and increased ADG
compared to the control group animals. In a similar
study done by Emanuelle et al. (1992) feeding the lambs
with inoculum-added dry forage improved the feed
consumption, BW gain, and feed conversion rate of the
animals.
Results of a study carried out by Nadeau et al. (2000)
revealed that bacterial inoculum and formic acid added
silage resulted in higher dry matter intake and improved
growth performance. In another study, it was indicated
that the addition of organic acid to the feed of steers
improved the ADG by 8.6% and enhanced the growth
performance (Martin et al., 1999).
The data, which were on the effects of direct-feed
microbials added to the feed, might be due to
differences in cultures used as direct-feed microbial as
well as variations in breed, age, gender, and
environmental conditions, as explained by Krehbiel et al.
(2003).
Results of the analysis of the serum samples obtained
from blood samples taken on day 28 and 56 of the study
are presented Table 5. Blood metabolite concentrations
were evaluated for nutritional status of Kivircik male
yearling lambs.
Regarding to the parameters concerning lipid
metabolism, the result of the present study was that the
Abas et al.: Organic Acid and Bacterial Direct-Feed Microbial
152
Table 4: Feed consumption and feed efficiency ratio of yearling lambs
Concentrate Forage Total
------------------------------------------------- ----------------------------------------------- --------------------------------------------------
Groups*
------------------------------------------------- ----------------------------------------------- --------------------------------------------------
COA DFM OA+ COA DFM OA+ COA DFM OA+
DFM DFM DFM
Feed Consumption (gram per day)
Day 0 - 28 1046 1056 1049 1052 975 1001 975 985 2020 2057 2023 2036
Day 29- 56 1107 1106 1107 1124 1049 1045 1049 1059 2156 2151 2155 2183
Day 0 - 56 1073 1078 1075 1083 1008 1021 1007 1018 2080 2099 2081 2101
Feed Efficiency (gain: feed)
Day 0 - 28 0.14 0.19 0.14 0.22 0.15 0.20 0.15 0.24 0.07 0.09 0.07 0.11
Day 29- 56 0.26 0.27 0.30 0.26 0.27 0.28 0.32 0.28 0.13 0.14 0.15 0.13
Day 0 - 56 0.20 0.23 0.22 0.24 0.21 0.24 0.24 0.26 0.10 0.12 0.12 0.13
* Groups: Control (C); Organic Acid (OA); Direct-fed microbials (DFM); Organic acid + Direct-fed microbials (OA+DFM)
Table 5: Effects of organic acid and/or direct-fed microbials on some blood parameters in yearling lambs
Groups SEM
-------------------------------------------------------------------------------------------------------------
COA DFM OA + DFM
----------------------------------------------------------------------- Initial ------------------------------------------------------------------------------
Albumin (g/L) 37.90 38.40 34.70 35.00 0.69
Total protein (g/L) 65.40 62.70 61.30 64.50 1.25
Urea (g/L) 0.26 0.20 0.27 0.28 0.02
Cholesterol (g/L) 0.86 0.91 0.82 0.78 0.02
Total lipid (g/L) 7.46 7.17 8.09 7.39 0.13
Ca (g/L) 0.09 0.09 0.10 0.10 0.003
Mg (mEq/L) 58.30 43.10 66.10 47.90 3.21
P (g/L) 0.05 0.04 0.05 0.05 0.002
i
Fe (mg/L) 1.20 1.20 1.30 1.30 0.53
Cu (mg/L) 0.70 0.71 0.61 0.78 0.03
Zn (mg/L) 0.61 0.54 0.64 0.53 0.02
------------------------------------------------------------------ Day 28 ------------------------------------------------------------------
Albumin (g/L) 40.50 38.10 41.90 42.30 0.82
Total protein (g/L) 62.10 55 65.40 62.40 2.15
Urea (g/L) 0.18 0.28 0.16 0.21 0.01*
b a b ab
Cholesterol (g/L) 7.69 5.95 8.24 7.42 0.03**
a b a ab
Total lipid (g/L) 7.11 7.39 7.33 6.91 0.11
Ca (g/L) 0.09 0.08 0.08 0.07 0.003
Mg (mEq/L) 53.4 70.5 68.0 52.1 2.50**
b a ab b
P (g/L) 0.04 0.05 0.05 0.04 0.001
i
Fe (mg/L) 1.4 1.3 1.4 1.2 0.05
Cu (mg/L) 0.98 0.86 0.96 0.93 0.03
Zn (mg/L) 0.85 0.69 0.91 0.77 0.02***
ab cabc
------------------------------------------------------------------ Day 56 ------------------------------------------------------------------
Albumin (g/L) 40.20 38.90 39.70 39.50 0.90
Total protein (g/L) 88.30 61.60 63.60 65.90 2.89***
a b b b
Urea (g/L) 0.20 0.24 0.19 0.22 0.01
Cholesterol (g/L) 9.60 7.23 8.41 7.98 0.22**
a b ab b
Total lipid (g/L) 8.02 7.20 7.33 6.70 0.15*
aab ab b
Ca (g/L) 0.10 0.08 0.08 0.09 0.003
Mg (mEq/L) 45.10 52.10 52.20 55 3.10
P (g/L) 0.04 0.03 0.04 0.03 0.002
i
Fe (mg/L) 1.80 1.70 1.80 1.80 0.06
Cu (mg/L) 0.93 0.94 0.88 0.84 0.02
Zn (mg/L) 0.64 0.84 0.86 0.85 0.03
b a a a
** Groups: Control (C); Organic Acid (OA); Direct-fed microbials (DFM); Organic acid + Direct-fed microbials (OA+DFM)
Means within sub rows with no common superscripts differ significantly (p<0.05). *p<0.05; ** p<0.01; *** p<0.001
a-c)
addition of the OA to the rations of the lambs significantly OA+DFM decreased the level of lipids significantly on
decreased the cholesterol level of the OA group animals day 56.
on days 28 and 56. On the other hand, the addition ofAlthough there are number of studies indicating that
organic acid and direct-feed microbial decreased the
total lipids relatively but not significantly. The addition ofdirect-feed microbials lower the blood cholesterol level
(Alp and Kahraman 1996; Lubbadeh et al., 1999), the
Abas et al.: Organic Acid and Bacterial Direct-Feed Microbial
153
results of the current study revealed that organic acidsConclusions: Results of the present study indicated that
decreased significantly the cholesterol level more thanaddition of organic acid or direct-feed microbial alone or
direct-feed microbials did. Lubbadeh et al., (1999)their combination to the rations of Kivircik Breed -male
brought up a possible explanation why the direct-feedyearling lambs increased body weight gain.
microbials decrease the level of cholesterol. MicrobialFurthermore, organic acid addition to the ration caused
feed additives reduce the absorption of lipid from the a decrease in blood cholesterol level while the addition
intestines by deconjugation; consequently it mayof direct-feed microbial caused an increase in blood Zn
decrease blood cholesterol. level.
The total protein, urea and albumin contents in blood are
indicators of protein metabolism in the organism. In this
study, the urea level of the yearling lambs in OA group
was found higher on day 28 than other groups (p<0.05).
This higher concentration of plasma urea was due to
increased quantity of ammonia absorbed from rumen
and circulating in the plasma because of the increasedin the lamb. Exp. Physiol., 76: 725-732.
feed consumption. Total protein level of the control group
animals was also found significantly higher than the
other groups at the end of the study period.
In one of the study, the effect of direct-feed microbials on
rumen metabolism and fattening performance in sheep
was observed and it was found that DFM did not change
the levels of some blood parameters including urea,
total proteins, and albumin. As a result, N metabolism
was not affected by yeast supplementation (Jouany et
al., 1998). In contrast to our result, Martin et al. (1999)
noted that blood urea concentration was decreased by
organic acid supplementation. The reduction in plasma
urea level for steers might have resulted from more
efficient utilization of dietary nitrogen. In addition,
Chiofalo et al. (2004) concluded that the application of
direct-feed microbial diet to growing Maltase goat kids
decreased blood total protein and urea levels. The
authors claimed that the lower content of urea could be
justified by better nutritional status.
During the trial, mineral metabolism was affected by
treatment. On day 28 of the present study; the serum Mg
level of the lambs in OA group was found significantly
higher than those of the control and OA+DFM groups. At
the end of this study (day 56), however, there was no
significant difference in Mg levels between the groups.
Another finding of the present study is that the addition
of direct-feed microbial to the ration caused and
increased serum Zn level (p<0.05).
The factors affecting mineral needs of animals,
inorganic composition of the body and tissues arekids. Reprod. Nutr. Dev., 44: 449-457.
determined by species of the animal, race, age, gender,
growth rate, health condition, nutritional condition,
endocrinological condition, season, and physiological
condition (lactation, pregnancy, dry period). In addition,
the mineral composition of the body and tissues depend
on levels of mineral and protein of the diet (i.e., amount,
chemical form, interaction between minerals) (Alp et al.,
2001; Kahraman et al., 2002). Abu-Damir et al., 1991,
reported that diet-induced change in blood acid-base
status was an important factor for mineral retention in
lambs and the rates of mineral retention were lowered
by acid diets in lambs.
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