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For Review OnlyEffect of flavor supplementation on growth performance, nutrient digestibility, blood profiles and carcass quality in
growing-finishing pigs
Journal: Canadian Journal of Animal Science
Manuscript ID CJAS-2019-0011.R3
Manuscript Type: Article
Date Submitted by the Author: 05-Jun-2019
Complete List of Authors: Ao, Xiang; Dankook University - Cheonan Campus, Animal Resource and Science; Tie Qi Li Shi Group. Co., Lei, Y.; DadHank Biotechnology CorporationKim, I.H.; Dankook University, Department of Animal Resource, and Science
Keywords: Anise, growth performance, growing-finishing pigs, nutrient digestibility, apple
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Running head: flavor in growing-finishing pigs
Effect of flavor supplementation on growth performance, nutrient digestibility,
blood profiles and carcass quality in growing-finishing pigs
Xiang Ao1, a, b, Yan Lei1, c, In Ho Kim*, a
a Department of Animal Resource & Science, Dankook University, #29 Anseodong,
Cheonan, Choognam, 330-714, Korea
b Tie Qi Li Shi Group. Co., Mianyang, Sichuan, 621006, P. R. China
c DadHank Biotechnology Corporation, Chengdu, Sichuan, 611130, P. R.China
* Corresponding author: Tel.: +82 41 550 3652; fax: +82 41 559 7881
E-mail address: [email protected] (I. H. Kim).
1These authors contributed equally to this work.
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ABSTRACT This study was conducted to evaluate the effect of different flavors
(apple and anise) supplementation on growth performance, nutrient digestibility,
blood profiles and carcass quality in growing-finishing pigs. A total of 96 growing
pigs [(Yorkshire×Landrace)×Duroc)] with an average BW of 28.2 ± 0.7 kg were
randomly allocated into 1 of the following 3 treatments: (1) CON, basal diet; (2) APF,
basal diet + 0.05% apple flavor; (3) ANF, basal diet + 0.05% anise flavor according to
their BW and sex in this 15-wk experiment. There were 8 replications (pens) per
treatment and 4 pigs per pen (2 barrows and 2 gilts). During wk 0-5, pigs fed ANF
diets had greater (p<0.05) average daily gain (ADG) and average daily feed intake
than those fed CON and APF diets. Dietary ANF treatment increased (p<0.05) ADG
during 0-15 wk compared with CON treatment. At the end of 5 wk, the apparent total
tract digestibility of nitrogen in ANF treatment was improved (p<0.05) compared
with that in CON treatment. Dietary treatments did not affect the studied traits of
carcass and meat quality. The inclusion of anise flavor increased ADG, but apple
flavor had no effect on growth performance in growing-finishing pigs.
Keywords: Anise, apple, growth performance, growing-finishing pigs, nutrient
digestibility
Abbreviations: ADFI, average daily feed intake; ADG, average daily gain; APF,
apple flavor; ANF, anise flavor; ATTD, apparent total tract digestibility; DM, dry
matter; G/F, gain: feed ratio; GE, gross energy; LM, longissimus muscle; N, nitrogen;
RBC, red blood cell; SEM, standard error means; WBC, white blood cell; WHC,
water holding capacity.
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Introduction
Nutrient intake is largely determined by voluntary feed intake, which is greatly
influenced by the flavor and taste of feed (Jacela et al., 2010). In order to stimulate
feed consumption, feed flavors are used to elevate the smell and taste of feed,
especially in post-weaning pigs (Roura et al., 2008). It is documented that diet aroma
and taste may be a factor that can affect the proportion of piglets consuming creep
feed before weaning (Sulabo et al., 2010). Therefore, previous studies indicated that
feed flavors were commonly included in weaning pigs’ diets to improve diet
acceptance and stimulate intake (McLaughlin et al., 1983; van Heugten et al., 2002;
Sulabo et al., 2010).
Previous study showed that animals with different species and different age prefer
different flavor (Oostindjer et al., 2010). Both milky flavor and fruit flavor could
enhance the feed intake of growing pigs (Lv et al., 2012). Additionally, previous
studies indicated that pre- and postnatal exposures to anise through the maternal diet
results in recognition by the offspring in humans, dogs and pigs (Schaal et al., 2000;
Hepper and Wells, 2006; Langendijk et al., 2007). Improvement in lymphocyte in
weanling pigs was observed in our previous study (Yan et al., 2011). However,
evidence of the potential effects of adding flavors to the growing-finishing pigs’ diet
on growth performance is limited. Therefore, the anise and apple flavors were
evaluated in this study. We hypothesized that the effect of flavor supplementation on
feed intake might exist in growing-finishing pigs. The objective of this study was to
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determine the impact of different flavors (apple and anise) on growth performance,
nutrient digestibility, blood profiles, carcass and meat quality in growing-finishing
pigs.
Materials and methods
Sources of flavors
The flavors used in this study were manufactured by DadHank Biotechnology
Corporation (Chengdu, China) as non-hygroscopic powder.
Animals, housing, and treatments
All animals received humane care as outlined in the Guide for the Care and Use of
Experimental Animals (Dankook University, Animal Care Committee). A total of 96
pigs [(Landrace × Yorkshire) × Duroc] with an average initial BW of 28.2 ± 0.7 kg
were assigned to 1 of 3 dietary treatments according to their sex and average pen BW
(8 replicates with 2 gilts and 2 barrows per replication pen). The treatments consisted
of (1) CON, basal diet; (2) APF, basal diet + 0.05% apple flavor; (3) ANF, basal diet
+ 0.05% anise flavor. The flavor was added at the expense of corn (0.05%). It was
premixed with corn and then mixed with the basal diet. Feed mixtures were pelleted
and formulated to provide all of the nutrients to meet or exceed NRC (2012)
requirements (Table 1). The experiment lasted for 15 wk.
All of the pigs were housed in an environmentally controlled facility with slatted
plastic flooring and a mechanical ventilation system. Each pen (1.8 × 1.8 m) was
provided with a stainless steel feeder and one nipple waterer, which allowed ad
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libitum access to feed and water throughout the experiment.
Experimental procedures, sampling, and analysis
The pigs were weighed individually at the beginning, wk 5, 10 and 15 of the
experiment. Feed consumption per pen was also assessed at the wk 5, 10 and 15 of the
experiment. The average daily gain (ADG), average daily feed intake (ADFI), and
gain-feed ratio (G:F) was calculated.
Chromic oxide (0.3%) was added to all the diets as an indigestible index for the
determination of apparent nutrient digestibility and the diet was provided to pigs for 7
days prior to fecal collection, chromium mixed feed were given from the beginning of
5, 10 and 15 wk and fecal were collected at the end of wk 5, 10 and 15. Fresh fecal
samples (at least 0.25 kg) were collected from 2 pigs (1 barrow and 1 gilt) during 1-d
period each pen via rectal massage, then pooled within the pens. All the feed and fecal
samples were stored at -20°C until further analysis. Before chemical analysis, fecal
samples were dried at 57°C for 72 h, after which they were ground to pass through a
1-mm screen. Experimental feeds were analyzed for nitrogen (N), calcium and
phosphorus and ether extract in accordance with AOAC procedures (2000). The
amino acid profile of diets was analyzed by HPLC (Hitachi L-8800 Amino Acid
Analyzer, Tokyo, Japan) as described by Lu et al. (2008). All samples were
hydrolyzed at 110°C for 24 h in 6 N HCl before analysis. Methionine was analyzed as
Met sulfone after cold performic acid oxidation overnight before hydrolysis. The
feces were analyzed for dry matter (DM), N and ether extract according to AOAC
(2000). Chromium was analyzed via UV absorption spectrophotometry (Shimadzu,
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UV-1201, Kyoto, Japan). The gross energy (GE) was determined by measuring the
heat of combustion in the samples using a Parr 6100 oxygen bomb calorimeter (Parr
instrument Co., Moline, IL). The apparent total tract digestibility (ATTD) of DM and
N was calculated using indirect-ratio methods. The GE in the feed and feces was
determined using a calorimeter (Mode1241, Parr Instrument Co., Illinois, USA). The
nutrient digestibility was calculated using the following formula: digestibility (%) ={1
– [(Nf × Cd) / (Nd × Cf)] } × 100 (Fenton et al., 1979), where Nf is the nutrient
concentration in feces (% DM), Nd the nutrient concentration in diet (% DM), Cd the
chromium concentration in diet (% DM), and Cf the chromium concentration in feces
(% DM).
On the final day of wk 5, 10 and 15, the same 2 pigs (1 barrow and 1 gilt) were
randomly selected from each pen and blood samples were collected via jugular vein
into K2 vacuum tubes (Becton Dickinson Vacutainer Systems, Franklin Lakes, NJ,
USA). The counts of red blood cell (RBC), white blood cell (WBC) and lymphocytes
in the blood were measured using the automatic blood analyzer (ADVIA 120, Bayer,
Tarrytown, NY, USA).
At the end of the experiment, all the pigs were transported (120 km, 2 h) to a
commercial abattoir for slaughter and were treated with conventional procedures. The
feed withdrawal time was 6 h. Treatment groups were kept separated in lairage to
avoid the effects of mixing (fighting) on ante-mortem muscle metabolism. They were
slaughtered by vertical exsanguinations after electrical stunning according to the
standard procedure under veterinarian control. The right leg was unshackled.
Carcasses were chilled at 2 °C for 24 h and a piece of the right loin was taken through
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a perpendicular cut between 10th and 11th ribs. Subjective meat color, marbling, and
firmness scores were evaluated by the same person according to National Pork
Producers Council (2000) standards. A visual evaluation for firmness of the loin was
assigned with a value of 1 (thin), 1.5 to 2.5 (average), or 3 (thick). Utilizing NPPC
visual reference standards with small modification, a subjective value for color, 1
(very pale) to 3 (very dark), was assigned to each carcass based on the color of the
lean tissue at the exposed longissimus muscle (LM) area, post blooming (Ao et al.,
2011). The NPPC visual reference standards for the subjective assessment of
intramuscular fat (IMF) content (marbling) were used to appropriately assign a
marbling score based on the amount of IMF interspersed within the lean tissue using a
scale of 1 (devoid) to 3 (excessive). The LM area was measured by tracing the LM
surface at the 10th rib, which also used the aforementioned digitizing arealine sensor.
Backfat thickness (at the area of the 10th/11th rib) was determined and the lean meat
percentage was measured using a real-time ultrasound instrument (Piglot 105; SFK
Technology, Herlev, Denmark). The pH in the middle of LM was measured in 24 h
post-mortem with an insertion glass electrode (Radiometer, Lyon, France) connected
to a pH-meter (NWKbinar pH, K-21, Landsberg, Germany). The electrode was
calibrated at 20 °C in buffers at pH value of 4.00 and 7.00. The water holding
capacity (WHC) was measured according to the methods described previously
(Kauffman et al., 1986). In brief, 0.2 g sample was pressed at 3,000 g for 3 min on a
125-mm-diameter filter paper. The areas of the pressed sample and expressed
moisture were delineated and then determined with a digitizing area-line sensor
(MT-10S; M.T. Precision Co. Ltd., Tokyo, Japan). A ratio of water: meat areas was
calculated, giving a measure of WHC (the smaller ratio indicate the higher the WHC).
Percentage of drip loss was determined according to Kuechenmeister et al. (2000).
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Briefly, meat samples were weighed, placed in a mesh bag, and then transferred to
nylon bag, and stored at chilling temperature for 24 h. Percentage of drip loss was
calculated by 100 x (initial weight – final weight of meat sample) / initial
(Kuechenmeister et al., 2000).
Statistical analysis
The data were analyzed by ANOVA using the GLM procedure of SAS (SAS Inst.
Inc., Cary, NC, USA, 2003) in a randomized complete block design with the pen
being considered as the experimental unit. Duncan’s multiple range test was used to
compare the means of the treatments. Variability in the data was expressed as the
standard error means (SEM) and a probability level of p<0.05 was considered to be
statistically significant.
Results
Growth performance
During wk 0-5, pigs fed ANF diets had greater (p<0.05) ADG and ADFI than those
fed CON and APF diets, while there was no difference (p>0.05) in G:F among
treatments (Table 2). During wk 6-10 and 11-15, dietary treatments did not influence
(p>0.05) ADG, ADFI or G:F. During wk 0-15, ADG in ANF treatment was increased
(p<0.05) compared with CON treatment, but no effect of dietary treatments was
observed (p>0.05) in ADFI or G:F.
Nutrient digestibility
At the end of wk 5, the ATTD of N was improved (p<0.05) by ANF treatment
compared with CON treatment, while there was no effect (p>0.05) on ATTD of DM
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or GE (Table 3). At the end of wk 10 and 15, dietary treatments did not affect (p>0.05)
ATTD of DM, N or GE.
Blood profiles and carcass quality
At the end of wk 5, 10 and 15, there was no difference (p>0.05) in the levels of
WBC, RBC or lymphocyte (Table 4). The carcass and meat quality (drip loss, pH,
WHC, subjective firmness, visual marbling, visual color, LMA, backfat thickness or
lean percentage) did not differ (p>0.05) among treatments (Table 5).
Discussion
Growth performance
Various studies observed the increase in ADFI by the feed flavor in weanling pigs
(van Heugten et al., 2002; Sulabo et al., 2010). The results were not always consistent.
Several studies demonstrated that dietary flavor supplementation did not affect ADG,
ADFI or G:F in weanling pigs (Kornegay et al., 1979; Mou et al., 2007). In our
previous study, the supplementation of anise flavor in sow and piglet diet improved
feed intake of sows and elevated growth performance of weaning pigs (Yan et al.,
2011). In the current study, the anise flavor supplementation improved ADG and
ADFI during growing phase (0-5 wk) and ADG throughout the entire experiment
(0-15 wk). In agreement with our results, the fruit flavor could increase ADG and
ADFI in growing pigs by elevating feed intake and reducing the time of feed intake
(Lv et al., 2012). On the contrary, feed flavor (0.05%) did not influence growth
performance in growing-finishing pigs (Thacker and Haq, 2009).
Nutrient digestibility
In our study, the ATTD of N was improved in growing pigs fed diets with anise
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flavor compared with those fed CON diet during the first five weeks (growing phase).
The similar results were observed in our previous study, which reported that the flavor
increased the nutrient digestibility of piglets in the first two weeks after weanling
(Yan et al., 2011). However, no difference was observed during the last ten weeks
(finishing phase) with flavor supplementation. Similarly, a study indicated that feed
flavor did not affect nutrient digestibility in growing-finishing pigs (Thacker and Haq,
2009). It is widely accepted that digestive capability of small intestine developed with
the increase of age, thus, different results may be due to the different growth phase. A
better digestion of dietary cereals and absorption of more nutrients were observed
with the maturation of gastrointestinal tract (Graham et al., 1986). Maturation of
gastrointestinal tract increased enzyme secretion, which increased nutrient
digestibility in pigs (Stewart et al., 2010). As it may explain why the flavor had
significant positive effect (no digestibility reference) on nutrient digestibility during
growing phase, but no significant difference was observed during finishing phase.
Blood profiles
Lymphocytes are both precursor cells of immunologic function as well as
regulators and effectors of immunity so that they play an important role in animal
immune system. There are limited studies conducted to evaluate the effects of flavor
on immune system of pigs. Improvement of immunity response in weanling pigs was
observed in our previous study (Yan et al., 2011). Notwithstanding, no effects on
blood profiles were observed in the current study, which may be attributed to more
developed digestive system, improved immunity and increased resistance to intestinal
disorders as pigs become older (Nousiainen and Setala, 1993). A study confirmed that
the largest immune organ in body was the gastrointestinal system and its associated
lymphoid, moreover, the maturation of gastrointestinal and optimal development of
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the immune system depended on the composition of the indigenous microflora (De
Vrese et al., 2007). In our study, growing-finishing pigs had much more developed
immune system than post-weanling pigs.
Carcass and meat quality
Meat color and LMA are considered as a determinant index deciding the
consumer's acceptance of the product. Dietary treatments did not affect carcass
quality in the present study, which was consistent with previous study (Thacker and
Haq, 2009). Previous study also demonstrated that formulating diets based on CP and
energy has no effects on ultimate muscle pH, drip loss percent, WHC, subjective color
scores, or firmness scores (Witte et al., 2000), which may explain the absence of
effect on carcass quality in our study. Meat pH is another carcass characteristic of
meat quality and affects the WHC of meat (Gou et al., 2002). With so minor effect on
the diet and considering that diet had no effect on growth performance, it is logical
that there is no effect on carcass and meat quality. The non-effect of flavor on WHC
in the current study may be attributed to the lack of pH discrepancy. To the best of our
knowledge, there are relatively limited data on the effect of flavors supplementation
on pigs, and further studies are needed to evaluate the effects of flavor on carcass
quality.
Conclusions
In summary, it can be concluded that anise flavor supplementation had minor effect
on growth performance and nutrient digestibility, but did not affect blood profiles,
carcass or meat quality in growing-finishing pigs.
Conflict of interests
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The authors declare that there is no conflict of interest.
Acknowledgement
This research was supported by a grant from the Bio-industry Technology
Development Program (315021-04) through the Korea Institute of Planning and
Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (iPET) and
KRIBB Research Initiative Program, Republic of Korea.
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Table 1. Diet composition (as-fed basis)Item Growing phase1 Finishing phase1
Ingredients, %Corn 59.25 67.85Soybean meal (CP, 48%) 26.55 17.84Rice bran 6.45 5.00Molasses 2.50 5.00Tallow 3.00 2.00Limestone 1.11 0.94Dicalcium phosphate 0.70 0.75NaCl 0.20 0.15L-Lys•HCl (78.8%) 0.02 0.30Vitamin premix2 0.12 0.07Trace mineral premix3 0.10 0.10
Analyzed compositionME, kcal/kg4 3,349 3,301CP, % 18.21 15.50Lys, % 0.97 0.79Met, % 0.31 0.21Ca, % 0.64 0.59Total P, % 0.52 0.52
1 Growing phase, 0-5 wk; finishing phase, 6-15 wk.2 Provided per kilogram of complete diet: 4,000 IU of vitamin A; 800 IU of vitamin D3; 17 IU of vitamin E; 2 mg of vitamin K; 4 mg of vitamin B2; 1 mg of vitamin B6; 16µg of vitamin B12; 11 mg of pantothenic acid; 20 mg of niacin and 0.02 mg of biotin.3 Provided per kilogram of complete diet: 220 mg of Cu; 175 mg of Fe; 191 mg of Zn; 89 mg of Mn; 0.3 mg of I; 0.5 mg of Co and 0.3 mg of Se.4 Calculated according to NRC [11].
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Table 2. Effects of different flavors on growth performance in growing-finishing pigs1
Item CON2 APF2 ANF2 SEM3 p-valueInitial BW, kg 28.14 28.27 28.19 0.7 0.52Final BW, kg 110.07 111.20 113.02 2.45 0.16wk 0 to 5ADG, g 646b 659b 706a 9 0.03ADFI, g 1,487b 1,510b 1,595a 14 0.04G:F 0.434 0.436 0.443 0.002 0.19
wk 6 to 10ADG, g 806 807 833 12 0.38ADFI, g 2,015 2,022 2,082 25 0.26G:F 0.400 0.399 0.400 0.002 0.83
wk 11 to 15ADG, g 889 905 906 15 0.25ADFI, g 2,622 2,618 2,653 28 0.60G:F 0.339 0.345 0.342 0.002 0.79
wk 0 to 15ADG, g 780b 790b 808a 10 0.04ADFI, g 2,035 2,022 2,110 26 0.14G:F 0.384 0.386 0.383 0.001 0.76
Note: Means within a column not sharing a lowercased italic letter differ significantly at the P < 0.05 level. 1 ADG mean represents 8 pens (n = 8/group) and feed consumption mean represents 8 pens (n = 8/group).2 CON, basal diet; APF, basal diet + 0.05% apple flavor; ANF, basal diet + 0.05% anise flavor.3Pooled standard error of the means.
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Table 3. Effects of different flavors on nutrient digestibility in growing-finishing pigs1
Item CON2 APF2 ANF2 SEM3 p-valuewk 5Dry matter 0.797 0.796 0.812 0.005 0.43Nitrogen 0.801b 0.809ab 0.820a 0.006 0.04Gross energy 0.766 0.780 0.784 0.011 0.23
wk 10Dry matter 0.765 0.767 0.779 0.005 0.16Nitrogen 0.762 0.770 0.781 0.007 0.12Gross energy 0.741 0.743 0.757 0.008 0.46
wk 15Dry matter 0.755 0.757 0.761 0.005 0.81Nitrogen 0.753 0.749 0.759 0.006 0.80Gross energy 0.729 0.727 0.738 0.007 0.52
Note: Means within a column not sharing a lowercased italic letter differ significantly at the P < 0.05 level. 1 Each mean represents 8 pens (n = 8/group).2 CON, basal diet; APF, basal diet + 0.05% apple flavor; ANF, basal diet + 0.05% anise flavor.3Pooled standard error of the means.
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Table 4. Effects of different flavors on blood profiles in growing-finishing pigs1
Item4 CON2 APF2 ANF2 SEM3 p-valuewk 5WBC, × 103/dl 18.19 19.24 20.07 0.47 0.18RBC, × 106/dl 5.57 5.74 6.22 0.34 0.54Lymphocyte, % 47.25 45.74 47.72 3.40 0.36
wk 10WBC, × 103/dl 21.69 22.08 23.17 0.47 0.09RBC, × 106/dl 6.59 6.64 6.82 0.34 0.94Lymphocyte, % 58.55 55.10 57.35 3.40 0.44
wk 15WBC, × 103/dl 24.17 23.10 24.89 1.42 0.37RBC, × 106/dl 6.67 7.15 7.44 0.31 0.11Lymphocyte, % 64.29 66.23 65.90 4.00 0.41
Note: Means within a column not sharing a lowercased italic letter differ significantly at the P < 0.05 level. 1 Each mean represents 8 pens (n = 8/group).2 CON, basal diet; APF, basal diet + 0.05% apple flavor; ANF, basal diet + 0.05% anise flavor.3Pooled standard error of the means.4 WBC, white blood cell; RBC, red blood cell; lymphocyte values are presented as a percentage of total WBC count.
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Table 5. Effects of different flavors on carcass quality in growing-finishing pigs1
Item4 CON2 APF2 ANF2 SEM3 p-valueDrip loss, %
d 1 2.92 3.12 2.76 0.12 0.70d 7 8.02 7.97 8.12 0.13 0.66
pH 5.57 5.62 5.69 0.02 0.11WHC, % 35.57 37.58 37.49 0.83 0.35Subjective evaluation
Firmness 1.97 1.77 1.94 0.06 0.49Marbling 2.15 2.14 2.21 0.05 0.98Color 2.35 2.56 2.44 0.07 0.85
LMA, cm2 44.67 44.26 47.58 1.07 0.37Backfat thickness, mm 25.7 25.9 25.5 0.31 0.75Lean percentage, % 55.6 55.7 57.0 0.40 0.43Note: Means within a column not sharing a lowercased italic letter differ significantly at the P < 0.05 level. 1 Each mean represents 8 pens (n = 8/group).2 CON, basal diet; APF, basal diet + 0.05% apple flavor; ANF, basal diet + 0.05% anise flavor.3Pooled standard error of the means.4 WHC, water hold capacity; LMA, longissimus muscle area.
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