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Article Citation:
Mohammad Mehdi Reyhani, Seyed Naser Mousavi, Ali Afsar and Hosein
Amirabdollahian
The effect of different physical form feeds and stocking density on performance
characteristics of carcass and immunity of broiler chickens
Journal of Research in Biology (2016) 6(7): 2110-2116
JournalofResearchinBiology
The effect of different physical form feeds and stocking density on
performance characteristics of carcass and immunity of broiler chickens
Keywords:
Broiler, density, mash, pellet, performance, stocking.
ABSTRACT:
The effect of different physical form feeds (pellet and mash) and stocking
density on the growth performance characteristics of carcass and immunity of Ross-
308 broiler chicks were studied. A total of 1800 one day-old ross-308 mixed-sex
broilers were used in a completely randomized design with six treatments and four
replicates of 75 birds each. They were arranged in a 2×3 factorial design: two feed
forms (mash and pellet) and three stocking density (10, 14 and 18 bird/m2
). Live body
Weight (LW), Weight Gain (WG), Feed Intake (FI) and Feed Conversion Ratio (FCR)
were measured periodically (0-10, 11-24 and 25-42 days). Carcass components and
litter quality were recorded at the end of the trial (day 45). Also antibody titer against
SRBC, and heterophil to lymphocyte ratio were measured in 45 day of age. The highest
body weight in grower period was observed in the pellet diet form (p<0.05). Also the
highest Body Weight (BW) and Body Weight Gain (BWG) in the starter period was
observed in the lowest stocking density treatment (10 bird/m2
) (p<0.05). Physical form
of diet had no significant effect on feed intake, BW and FCR throughout the periods.
However the broiler feed intake was significantly influenced by stocking density and a
decrease in the high stocking density group (18 bird/m2
). Different types of feed and
stocking density had no significant effect on carcass characteristics, antibody titer and
H:L ratio. Use of pellet form diet and high stocking density treatment (18 bird/m2
)
significantly increased broiler litter moisture (P<0.05).
2110-2116 | JRB | 2016 | Vol 6 | No 7
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
licenses/by/4.0), which gives permission for unrestricted use, non-commercial, distribution and
reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com
Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Mohammad Mehdi
Reyhani1
,
Seyed Naser Mousavi1
,
Ali Afsar2
and
Hosein Amirabdollahian3
Institution:
1. M.Sc. in Animal
Nutrition, Islamic Azad
University, Varamin Branch.
Iran.
2. Ph.D in animal nutrition,
Islamic Azad University,
Varamin Branch. Iran.
3. M.Sc. in Poultry Breeding
and Production
Management, Islamic Azad
University, Garmsar Branch.
Iran.
Corresponding author:
Mohammad Mehdi
Reyhani
Email Id:
Web Address:
http://jresearchbiology.com/
documents/RA0623.pdf
Dates:
Received: 27 Aug 2016 Accepted: 12 Sep 2016 Published: 12 Oct 2016
Original Research
Journal of Research in Biology
An International Scientific Research Journal
ISSN No: Print: 2231 –6280; Online: 2231- 6299
INTRODUCTION
Feed constitutes around 60-70 percent of the total
cost of broiler production and different commercial feed
form (mash, crumble and pellet) are delivered in poultry
industry (Banerjee, 1998). The effective utilization of
feed is critical in broiler production. Mash is a finely
ground and blended feed that gives more prominent
unification of growth and is more economical. Pellet is a
type of complete feed that is compacted and expelled to
around 1.8 inch in diameter and 1.4 inch in long
(Banerjee, 1998). Pelleting of feed has a few advantages:
expanding the mass thickness of feed and enhancing feed
flow capacity. Be that as it may, by and large, pellet or
crumble costs somewhat more than a similar proportion
in mash form.
Contrasted with mash, pellets enhance bird
performance by diminishing feed wastage, mitigating
selective feeding, destroying pathogens, enhancing
palatability and expanding nutrient digestibility. Some
disadvantages of the pellet are pelleting costs about 10%
more than producing mash feed and the litter moisture
increase when the feed pellets are used (Jahan et al.,
2006). Previously, Mingbin et al., (2015) reported higher
average daily gain and average daily feed intake for birds
fed with the crumble-pellet diets than for those fed the
mash diets during starter, grower and the entire
experimental period. Chewning et al. (2012) announced
that broilers fed pelleted diets have higher BW and
enhanced feed conversion than those fed mash feed.
However, feeding pelleted apportions isn't sufficient to
guarantee upgraded performance of poultry. The nature
of pellets must be considered too.
Stocking density assumes a critical part in broiler
production. Higher mortality, bring down meat
production, more noteworthy rate of leg disorders and
cannibalism happen at higher stocking densities in
broilers. Negative impacts of high stocking densities on
broiler performance have been accounted for in past
investigations (Dozier et al., 2005 and 2006).
A lessening in the wind stream at the bird level, which
happened at the high stocking densities, could diminish
the dispersal of body heat to the air. A lessening in
access to water and feed, enhancement ammonia and an
unfavorable air quality as a result of reduced air
exchange are different components that could adversely
impact bird performance. (Feddes et al., 2002). Also,
high stocking densities make a strees condition for bird
and may decrease immune function, too. Eriflir et al.
(2002) detailed that there was a critical difference in
immune response with an expansion in stocking density
in Japanese quails.
The objective of this study was to investigate the
effects of feed form (mash and pellet) and different
stocking density (10, 14 and 18 bird/m2
) on the
performance, immunity and carcass characteristics of the
broiler chickens.
MATERIALS AND METHODS
The experiment was divided into three phases,
i.e., starter (d 1 to 10), grower (d 11 to 24) and finisher (d
25 to 45). A sum of 1800 one day-old Ross 308 blended
sex broiler were utilized as a part of a completely
randomized design with six treatments and four
duplicates of 75 birds each. They were arranged in a 2×3
factorial design: two feed forms (mash and pellet) and
three stocking density (10, 14 and 18 bird/m2
). Initial
room temperature was 34°C and was then diminished by
2°C every week until the point that a temperature of
26°C was accomplished. Feed and water were provided
ad libitum. On day 1, 11, 25 and 45, chicks were
weighed by pen and feed consumption was recorded.
Body Weight (BW), Body Weight Gain (BWG), Feed
Intake (FI) and Feed Conversion Ratio (FCR) including
mortality weight, were calculated for each phase. The
compositions of the experimental diets are presented in
Table 1.
On day 45, four birds per treatment were
randomly selected, weighed and then killed. After
Reyhani et al., 2016
2111 Journal of Research in Biology (2016) 6(7): 2110-2116
removal of feathers, feet and head, carcass yield was
determined. Cut-up parts such as thigh, breast, thigh,
back neck and abdominal fat were weighed. Meanwhile,
gizzard, small intestine, heart, liver, spleen and bursa of
fabricius were removed and weighed. Blood samples
were collected at the 45th
day of the study. A total of 8
arbitrarily chosen chickens from each group were
tenderly expelled from their rooms and blood samples
(0.5 ml) were taken into EDTA tubes for heterophil and
lymphocyte counts. Blood smears were prepared
utilizing May-Grunwald-Giemsa stain, and heterophil
and lymphocytes were checked to total of 60 cells.
(Gross and Siegel, 1983). The serum of blood samples in
the second tube was separated and used to measure
antibody titer against SRBC by ELISA. Antibody titer
data were logarithmically transformed before analyses.
For the litter quality measurement, litter samples were
collected from four points of each replicate and their
moisture were measured with oven dray method (1050C
for 24h). Data were subjected to 2-way ANOVA by
using the GLM procedure of SAS (SAS Institute, 2005).
RESULTS AND DISCUSSION
The results of broiler performance as affected by
physical form of feed and stocking density are shown in
Table 2. Body weight was significantly affected by
physical form and stocking density (p<0.05). The highest
body weight in grower period was observed in pellet diet
form (p<0.05). Also, the highest Body Weight (BW) and
Journal of Research in Biology (2016) 6(7): 2110-2116 2112
Reyhani et al., 2016
Ingredients Starter (0-10) Grower (11-24) Finisher (25-42)S. No
Maize 48.53 48.94 52.931
Soybean meal 36.5 30.16 25.32
Wheat 8 15 153
Soybean oil 1.86 1.56 2.524
CaCO3 1.74 1.44 1.435
Monocalcium phosphate 1.53 1.35 1.276
Common salt 0.32 0.27 0.2377
NaHCO3 0.1 0.1 0.158
DL-Methionine 0.32 0.25 0.239
Lysine hydrochloride 0.22 0.16 0.17410
Threonine 0.09 0.06 0.0611
Vitamin1
and Mineral2
Premix
0.3 0.3 0.312
Choline chloride 0.1 0.08 0.0713
Plate binder 0.2 0.15 0.1514
Coccidio acetate 0.025 0.025 0.02515
Sorbatox 0.05 0.05 0.0516
Lupeol 0.1 0.1 0.117
Digestrom 0.012 0.012 0.01218
Total 100 100 100
Table 1. Composition of the experimental diets
1. The vitamin premix supplied the following per kilogram of diet: vitamin A (retinyl acetate), 8,000 IU; vitamin D3,
1,000 IU; vitamin E (dl-α tocopherol), 30 IU; vitamin K3, 2.5 mg; vitamin B1, 2 mg; vitamin B2, 5 mg; vitamin B6, 2
mg; vitamin B12, 0.01 mg; niacin, 30 mg; d-biotin, 0.045 mg; vitamin C, 50 mg; d-pantothenate, 8 mg, folic acid, 0.5
mg.
2. The mineral premix supplied the following per kilogram of diet: Mn, 70 mg; Fe, 35 mg; Zn, 70 mg; Cu, 8 mg;
I, 1 mg, Se, 0.25 mg; Co, 0.2 mg
Reyhani et al., 2016
2113 Journal of Research in Biology (2016) 6(7): 2110-2116
BodyWeight(BW)BodyWeightGain(BWG)FeedintakeFeedconversionratio
0-1111-2525-450-1111-2525-451-450-1111-2525-451-450-1111-2525-451-45
Feed
Form
Mash260.5764.7b
2523.2221.1504.21758.52485.7314.1806.83571.54692.61.41.62.021.8
Pellet269.8816.6a
2608.7230.3547.51791.12569.1317.5820.835654703.41.31.51.91.8
SEM4.514.549.24.215.552.549.26.421113.2111.40.030.040.030.02
Stoking
Density
10276.0a
7872573.1237.1a
5111786.12537.1341.8b
845.13522.64692.71.41.61.91.8
14269.87ab
783.82533.3229.5ab
5141748.12491.7323.3b
806.23519.54648.81.41.52.011.8
18249.6a
801.22591.5210.5b
552.71790.22553.5282.4b
807.13662.74752.41.31.42.041.8
SEM5.517.860.35.119.0864.360.27.825.7138.7136.40.030.050.040.02
Mash10267.7752.72502.5228.54851749.724.69346.6808.73407.54563.21.51.61.91.8
Pellet10284.2821.22643.7245.75371822.52605.2337.1847.53637.74822.31.31.51.91.8
Mash14271.0739.72509.7231.07468.717702469312.1792.53666.74771.11.31.62.061.9
Pellet14268.78282557228.1559.21726.22513.7334.38203372.24526.31.41.41.91.8
Mash18242.7801.72557.5203.85591755.72518.6283.6819.23640.24743.21.31.52.061.8
Pellet18256.5800.72625.5217.1546.51824.72588.3281.37953685.24761.61.31.42.011.8
SEM7.825.285.37.2826.990.985.21136.3196.21930.050.080.050.04
Table2.Effectofdifferentfeedform(mashandpellet)andstockingdensityonbroilerperformance
a,bMeansincolumnnotsharingacommonsuperscriptaresignificantlydifferent(P<0.05)
Body Weight Gain (BWG) in starter period was
observed in the lowest stocking density treatment (10
bird/m2
) (p<0.05). Physical form of diet had no
significant effect on feed intake, BW and Feed
Conversion Ratio (FCR) throughout the periods.
However the broiler feed intake was significantly
influenced by stocking density and the decrease in the
high stocking density group (18 bird/m2
).
Similarly, Mingbin et al. (2015) indicated that
the use of pellet form in broiler diet had positive effects
on body weight. However, the pellet form diet had no
significant effect on other performance parameters
(BWG, feed intake and FCR). These results were in
agreement with Salari et al. (2006), who found that the
form of diet and particle size had no significant effect on
weight gain and dry matter intake. This observation
suggested that high pellet quality may be necessary to
fully obtain benefits of pelleting. Birds at normal
stocking density (10 birds/m2
) resulted in better body
weight in starter period compared with the high stocking
density (18 birds/m2
). This indicates to a greater degree
of stress on the performance. However, other
performance parameters in other periods weren’t affected
by different stocking density. Additionally, Buijs et al.
(2009) detailed that at body weight 39 days of age was
not significant between birds raised at various stocking
densities (6, 15, 23, 33, 35, 41, 47 and 56 kg /m2
).
Interestingly, Houshmand et al. (2012) revealed that
amid the growing stage (22-42 days) broilers raised at a
high density had a inferior FCR contrasted and birds
housed at normal density.
As shown in Table 3, all carcass parameters
(carcass, breast, thigh, back and neck), abdominal fat,
some inner organ weight (small intestine, heart, liver,
spleen and bursa of Fabricius) except for the gizzard
percent weight were not significantly (p>0.05)
influenced by the form of the diet and different stocking
density. Use of mash diet significantly increased the
gizzard percent weight (p<0.05). A similar observation
was also recorded by Ravindran and Thomas (2004) who
reported that carcass characteristics weren’t affected by
stocking density. Previously, Ghorbani et al. (2012)
Journal of Research in Biology (2016) 6(7): 2110-2116 2114
Reyhani et al., 2016
Treatment Breast Thigh Back Gizzard
Small
intestine
Abdominal
fat
Heart Liver Spleen Bursa
Feed form Mash 20.19 20.66 16.03 1.92a
1.78 1.45 2.02 2.31 0.1 0.11
Pellet 20.53 20.33 15.97 1.65b
1.89 1.48 0.53 2.24 0.1 0.11
SEM 0.3 0.1 0.3 0.04 0.03 0.07 1.00 0.03 0.03 0
Stock
density
10 20.45 20.57 15.96 1.75 1.88 1.44 0.66 2.21 0.09 0.12
14 20.55 20.50 15.49 1.80 1.80 1.46 2.7 2.29 0.1 0.11
18 20.07 20.42 16.54 1.80 1.82 1.5 0.4 2.32 0.1 0.11
SEM 0.3 0.2 0.4 0.05 0.03 0.08 1.2 0.04 0.0 0.0
Interaction
Mash 10 20.1 20.70 15.89 1.89 1.87 1.44 0.72 2.28 0.10 0.12
Pellet 10 20.8 20.44 16.04 1.61 1.89 1.43 0.60 2.15 0.09 0.11
Mash 14 20.93 21.04 15.65 2.01 1.71 1.38 4.96 2.36 0.09 0.11
Pellet 14 20.18 19.96 15.33 1.59 1.88 1.53 0.48 2.23 0.10 0.11
Mash 18 19.53 20.24 16.56 1.85 1.75 1.51 0.37 2.28 0.10 0.11
Pellet 18 20.60 20.59 16.53 1.75 1.89 1.49 0.52 2.35 0.10 0.11
SEM 0.54 0.29 0.61 0.07 0.05 0.12 1.76 0.06 0.005 0.001
a, b Means in column not sharing a common superscript are significantly different (p<0.05)
Table 3. Effect of different feed form (mash and pellet) and stocking density on carcass characteristics and
some inner organ weight (%)
reported that the relative weight of gizzard increases by
mash diet in compared with pellet form diet. The
increase in relative weight of gizzard may enhance the
digestive capacity of broilers.
Mean antibody titers to sheep red blood cell, H:
L ratio and litter moisture are presented in Table 4.
Different physical feed type and stocking density had no
significant effect on the antibody titers to sheep red
blood and H/L ratio in the broiler chicks. However the
litter moisture was significantly influenced by treatments
(p<0.05). Use of pellet form diet and high stocking
density treatment (18 bird/m2
) significantly increased
broiler litter moisture (P<0.05). The H:L proportion is a
typical marker of stress in poultry (Heckert et al. 2002),
and the blood leukocyte profile is impacted by stress.
Decrease in the quantities of lymphocytes and monocytes
and improvement in the quantities of heterophils, which
prompts a higher H:L proportion, have been accounted
for stressed animals (Stevenson and Taylor, 1988).
Previously, Houshmand et al. (2012) reported that
density had no significant effect on the heterophil:
lymphocyte ratio too. In high stocking density, especially
in hot and wet weather, litter moisture also gets increased
and causes the increase in bacterial activity and ammonia
production. Increased mortality can be explained by
decreased animal welfare, such as bad air and litter
quality, poor immune response, and poor feed intake.
Litter quality has a large effect on dermatitis (Haslam et
al., 2006).
CONCLUSIONS
Results showed that the use of pellet form diet
and normal stocking density (10 bird/m2
) can improve
broiler body weight and litter quality.
ACKNOWLEDGMENTS
We sincerely thank the members of animal
science department of Islamic Azad University, Varamin
branch, Tehran, Iran.
REFERENCES
Banerjee GC. (1998). Poultry. 3rd
edition, Oxford and
IBH Publishing Co. Pvt. Ltd. Calcutta. 121p.
Buijs S, Keeling L, Rettenbacher S, Van Poucke E
and Tuyttens FAM. (2009). Stocking density effects on
broiler welfare: identifying sensitive ranges for different
Reyhani et al., 2016
2115 Journal of Research in Biology (2016) 6(7): 2110-2116
Treatment SRBC H/L (%) Litter moisture (%)S. No
Feed form Mash 9.52 0.31 9.55b1
Pellet 9.47 0.31 12.12a2
SEM 0.5 0.00 0.53
Stock density4
10 10.12 0.318 8.66b
5
14 9.20 0.306 8.11b
6
18 9.16 0.310 15.74a
7
SEM 0.7 0.008 0.78
Interaction9
Mash 10 10.91 0.32 9.26b
10
Pellet 10 9.33 0.31 8.07b
11
Mash 14 9.08 0.30 7.91b
12
Pellet 14 9.33 0.30 8.32b
13
Mash 18 8.58 0.30 11.5b
14
Pellet 18 9.75 0.32 19.98a
15
SEM 1.03 0.01 0.916
Table 4. Effect of different feed form (mash and pellet) and stocking density on immunity and litter quality
a, b Means in column not sharing a common superscript are significantly different (p<0.05)
indicators. Poultry Science, 88(8): 1536-1543.
Chewning CG, Stark CR and Brake J. (2012). Effects of
particle size and feed form on broiler performance. The Journal
of Applied Poultry Research, 21(4): 830–837.
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HA, Branton SL and Roush WB. (2006). Stocking density
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Dozier WA, Thaxton JP, Branton SL, Morgan GW, Miles
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Eriflir M. (2002). Yerleflim sıklıı arttırılan bıldırcınların
(Coturnix coturnix japonica) bazı biyokimyasal kan
parametrelerindeki deoifliklikler. Turkish Journal of Veterinary
Animal Science, 26: 491-496.
Feddes JJ Emmanuel EJ and Zuidhof MJ. (2002). Broiler
performance, body weight variance, feed and water intake, and
carcass quality at different stocking densities. Poultry Science,
81(6): 774–779.
Ghobadi Z and Karimi A. (2012). Effect of feed processing
and enzyme supplementation of wheat-based diets on
performance of broiler chicks. Journal of Applied Animal
Research, 40(3): 260 266.
Gross WB and Siegel HS. (1983). Evaluation of the
heterophil/lymphocyte ratio as a measure of stress in chickens.
Avian Disease, 27(4): 972-979.
Haslam SM, Brown SN, Wilkins LJ Kestin SC, Warriss PD
and Nicol CJ. (2006). Preliminary study to examine the utility
of using foot burn or hock burn to assess aspects of housing
conditions for broiler chicken. British Poultry Science, 47(1):
13–18.
Heckert RA, Estevez I, Russek-Cohen E and Pettit-Riley R.
(2002). Effects of density and perch availability on the immune
status of broilers. Poultry Science, 81(4): 451–457.
Houshmand M, Azhar K, Zulkifli I, Bejo MH and Kamyab
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density on performance, immunity and stress indicators of
broilers. Poultry Science, 91(2): 393-401.
Jahan MS, Asaduzzaman M and Sarkar AK. (2006).
Performance of broiler fed on mash, pellet and crumble.
International Journal of Poultry Science, 5(3): 265–70.
Mingbin Lv, Lei Yan, Zhengguo W, Sha An, Miaomiao Wu
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digestive tract development of broilers. Animal Nutrition,
1(3): 252–256.
Ravindran V and Thomas DV. (2004). Performance and
welfare of broilers as affected by stocking density and in-feed
antibiotic supplementation. Proceedings of the 16th
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Poultry Science Symposium, Sydney, New-South Wales,
Australia, 9-11 ,135-138.
Salari S, Kermanshahi H and Nasiri MH. (2006). Effect of
sodium bentonite and comparison of pellet vs mash on
performance of broiler chickens. International Journal of
Poultry Science, 5(1): 31–4.
SAS Institute. (2005). User’s Guide. Version 9. SAS Inst. Inc.,
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Stevenson JR and Taylor R. (1988). Effects of glucocorticoid
and antiglucocorticoid hormones on leukocyte numbers and
function. International Journal of Immunopharmacology,
10(1): 1–6.
Journal of Research in Biology (2016) 6(7): 2110-2116 2116
Reyhani et al., 2016
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The effect of different physical form feeds and stocking density on performance characteristics of carcass and immunity of broiler chickens

  • 1. Article Citation: Mohammad Mehdi Reyhani, Seyed Naser Mousavi, Ali Afsar and Hosein Amirabdollahian The effect of different physical form feeds and stocking density on performance characteristics of carcass and immunity of broiler chickens Journal of Research in Biology (2016) 6(7): 2110-2116 JournalofResearchinBiology The effect of different physical form feeds and stocking density on performance characteristics of carcass and immunity of broiler chickens Keywords: Broiler, density, mash, pellet, performance, stocking. ABSTRACT: The effect of different physical form feeds (pellet and mash) and stocking density on the growth performance characteristics of carcass and immunity of Ross- 308 broiler chicks were studied. A total of 1800 one day-old ross-308 mixed-sex broilers were used in a completely randomized design with six treatments and four replicates of 75 birds each. They were arranged in a 2×3 factorial design: two feed forms (mash and pellet) and three stocking density (10, 14 and 18 bird/m2 ). Live body Weight (LW), Weight Gain (WG), Feed Intake (FI) and Feed Conversion Ratio (FCR) were measured periodically (0-10, 11-24 and 25-42 days). Carcass components and litter quality were recorded at the end of the trial (day 45). Also antibody titer against SRBC, and heterophil to lymphocyte ratio were measured in 45 day of age. The highest body weight in grower period was observed in the pellet diet form (p<0.05). Also the highest Body Weight (BW) and Body Weight Gain (BWG) in the starter period was observed in the lowest stocking density treatment (10 bird/m2 ) (p<0.05). Physical form of diet had no significant effect on feed intake, BW and FCR throughout the periods. However the broiler feed intake was significantly influenced by stocking density and a decrease in the high stocking density group (18 bird/m2 ). Different types of feed and stocking density had no significant effect on carcass characteristics, antibody titer and H:L ratio. Use of pellet form diet and high stocking density treatment (18 bird/m2 ) significantly increased broiler litter moisture (P<0.05). 2110-2116 | JRB | 2016 | Vol 6 | No 7 This article is governed by the Creative Commons Attribution License (http://creativecommons.org/ licenses/by/4.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited. www.jresearchbiology.com Journal of Research in Biology An International Scientific Research Journal Authors: Mohammad Mehdi Reyhani1 , Seyed Naser Mousavi1 , Ali Afsar2 and Hosein Amirabdollahian3 Institution: 1. M.Sc. in Animal Nutrition, Islamic Azad University, Varamin Branch. Iran. 2. Ph.D in animal nutrition, Islamic Azad University, Varamin Branch. Iran. 3. M.Sc. in Poultry Breeding and Production Management, Islamic Azad University, Garmsar Branch. Iran. Corresponding author: Mohammad Mehdi Reyhani Email Id: Web Address: http://jresearchbiology.com/ documents/RA0623.pdf Dates: Received: 27 Aug 2016 Accepted: 12 Sep 2016 Published: 12 Oct 2016 Original Research Journal of Research in Biology An International Scientific Research Journal ISSN No: Print: 2231 –6280; Online: 2231- 6299
  • 2. INTRODUCTION Feed constitutes around 60-70 percent of the total cost of broiler production and different commercial feed form (mash, crumble and pellet) are delivered in poultry industry (Banerjee, 1998). The effective utilization of feed is critical in broiler production. Mash is a finely ground and blended feed that gives more prominent unification of growth and is more economical. Pellet is a type of complete feed that is compacted and expelled to around 1.8 inch in diameter and 1.4 inch in long (Banerjee, 1998). Pelleting of feed has a few advantages: expanding the mass thickness of feed and enhancing feed flow capacity. Be that as it may, by and large, pellet or crumble costs somewhat more than a similar proportion in mash form. Contrasted with mash, pellets enhance bird performance by diminishing feed wastage, mitigating selective feeding, destroying pathogens, enhancing palatability and expanding nutrient digestibility. Some disadvantages of the pellet are pelleting costs about 10% more than producing mash feed and the litter moisture increase when the feed pellets are used (Jahan et al., 2006). Previously, Mingbin et al., (2015) reported higher average daily gain and average daily feed intake for birds fed with the crumble-pellet diets than for those fed the mash diets during starter, grower and the entire experimental period. Chewning et al. (2012) announced that broilers fed pelleted diets have higher BW and enhanced feed conversion than those fed mash feed. However, feeding pelleted apportions isn't sufficient to guarantee upgraded performance of poultry. The nature of pellets must be considered too. Stocking density assumes a critical part in broiler production. Higher mortality, bring down meat production, more noteworthy rate of leg disorders and cannibalism happen at higher stocking densities in broilers. Negative impacts of high stocking densities on broiler performance have been accounted for in past investigations (Dozier et al., 2005 and 2006). A lessening in the wind stream at the bird level, which happened at the high stocking densities, could diminish the dispersal of body heat to the air. A lessening in access to water and feed, enhancement ammonia and an unfavorable air quality as a result of reduced air exchange are different components that could adversely impact bird performance. (Feddes et al., 2002). Also, high stocking densities make a strees condition for bird and may decrease immune function, too. Eriflir et al. (2002) detailed that there was a critical difference in immune response with an expansion in stocking density in Japanese quails. The objective of this study was to investigate the effects of feed form (mash and pellet) and different stocking density (10, 14 and 18 bird/m2 ) on the performance, immunity and carcass characteristics of the broiler chickens. MATERIALS AND METHODS The experiment was divided into three phases, i.e., starter (d 1 to 10), grower (d 11 to 24) and finisher (d 25 to 45). A sum of 1800 one day-old Ross 308 blended sex broiler were utilized as a part of a completely randomized design with six treatments and four duplicates of 75 birds each. They were arranged in a 2×3 factorial design: two feed forms (mash and pellet) and three stocking density (10, 14 and 18 bird/m2 ). Initial room temperature was 34°C and was then diminished by 2°C every week until the point that a temperature of 26°C was accomplished. Feed and water were provided ad libitum. On day 1, 11, 25 and 45, chicks were weighed by pen and feed consumption was recorded. Body Weight (BW), Body Weight Gain (BWG), Feed Intake (FI) and Feed Conversion Ratio (FCR) including mortality weight, were calculated for each phase. The compositions of the experimental diets are presented in Table 1. On day 45, four birds per treatment were randomly selected, weighed and then killed. After Reyhani et al., 2016 2111 Journal of Research in Biology (2016) 6(7): 2110-2116
  • 3. removal of feathers, feet and head, carcass yield was determined. Cut-up parts such as thigh, breast, thigh, back neck and abdominal fat were weighed. Meanwhile, gizzard, small intestine, heart, liver, spleen and bursa of fabricius were removed and weighed. Blood samples were collected at the 45th day of the study. A total of 8 arbitrarily chosen chickens from each group were tenderly expelled from their rooms and blood samples (0.5 ml) were taken into EDTA tubes for heterophil and lymphocyte counts. Blood smears were prepared utilizing May-Grunwald-Giemsa stain, and heterophil and lymphocytes were checked to total of 60 cells. (Gross and Siegel, 1983). The serum of blood samples in the second tube was separated and used to measure antibody titer against SRBC by ELISA. Antibody titer data were logarithmically transformed before analyses. For the litter quality measurement, litter samples were collected from four points of each replicate and their moisture were measured with oven dray method (1050C for 24h). Data were subjected to 2-way ANOVA by using the GLM procedure of SAS (SAS Institute, 2005). RESULTS AND DISCUSSION The results of broiler performance as affected by physical form of feed and stocking density are shown in Table 2. Body weight was significantly affected by physical form and stocking density (p<0.05). The highest body weight in grower period was observed in pellet diet form (p<0.05). Also, the highest Body Weight (BW) and Journal of Research in Biology (2016) 6(7): 2110-2116 2112 Reyhani et al., 2016 Ingredients Starter (0-10) Grower (11-24) Finisher (25-42)S. No Maize 48.53 48.94 52.931 Soybean meal 36.5 30.16 25.32 Wheat 8 15 153 Soybean oil 1.86 1.56 2.524 CaCO3 1.74 1.44 1.435 Monocalcium phosphate 1.53 1.35 1.276 Common salt 0.32 0.27 0.2377 NaHCO3 0.1 0.1 0.158 DL-Methionine 0.32 0.25 0.239 Lysine hydrochloride 0.22 0.16 0.17410 Threonine 0.09 0.06 0.0611 Vitamin1 and Mineral2 Premix 0.3 0.3 0.312 Choline chloride 0.1 0.08 0.0713 Plate binder 0.2 0.15 0.1514 Coccidio acetate 0.025 0.025 0.02515 Sorbatox 0.05 0.05 0.0516 Lupeol 0.1 0.1 0.117 Digestrom 0.012 0.012 0.01218 Total 100 100 100 Table 1. Composition of the experimental diets 1. The vitamin premix supplied the following per kilogram of diet: vitamin A (retinyl acetate), 8,000 IU; vitamin D3, 1,000 IU; vitamin E (dl-α tocopherol), 30 IU; vitamin K3, 2.5 mg; vitamin B1, 2 mg; vitamin B2, 5 mg; vitamin B6, 2 mg; vitamin B12, 0.01 mg; niacin, 30 mg; d-biotin, 0.045 mg; vitamin C, 50 mg; d-pantothenate, 8 mg, folic acid, 0.5 mg. 2. The mineral premix supplied the following per kilogram of diet: Mn, 70 mg; Fe, 35 mg; Zn, 70 mg; Cu, 8 mg; I, 1 mg, Se, 0.25 mg; Co, 0.2 mg
  • 4. Reyhani et al., 2016 2113 Journal of Research in Biology (2016) 6(7): 2110-2116 BodyWeight(BW)BodyWeightGain(BWG)FeedintakeFeedconversionratio 0-1111-2525-450-1111-2525-451-450-1111-2525-451-450-1111-2525-451-45 Feed Form Mash260.5764.7b 2523.2221.1504.21758.52485.7314.1806.83571.54692.61.41.62.021.8 Pellet269.8816.6a 2608.7230.3547.51791.12569.1317.5820.835654703.41.31.51.91.8 SEM4.514.549.24.215.552.549.26.421113.2111.40.030.040.030.02 Stoking Density 10276.0a 7872573.1237.1a 5111786.12537.1341.8b 845.13522.64692.71.41.61.91.8 14269.87ab 783.82533.3229.5ab 5141748.12491.7323.3b 806.23519.54648.81.41.52.011.8 18249.6a 801.22591.5210.5b 552.71790.22553.5282.4b 807.13662.74752.41.31.42.041.8 SEM5.517.860.35.119.0864.360.27.825.7138.7136.40.030.050.040.02 Mash10267.7752.72502.5228.54851749.724.69346.6808.73407.54563.21.51.61.91.8 Pellet10284.2821.22643.7245.75371822.52605.2337.1847.53637.74822.31.31.51.91.8 Mash14271.0739.72509.7231.07468.717702469312.1792.53666.74771.11.31.62.061.9 Pellet14268.78282557228.1559.21726.22513.7334.38203372.24526.31.41.41.91.8 Mash18242.7801.72557.5203.85591755.72518.6283.6819.23640.24743.21.31.52.061.8 Pellet18256.5800.72625.5217.1546.51824.72588.3281.37953685.24761.61.31.42.011.8 SEM7.825.285.37.2826.990.985.21136.3196.21930.050.080.050.04 Table2.Effectofdifferentfeedform(mashandpellet)andstockingdensityonbroilerperformance a,bMeansincolumnnotsharingacommonsuperscriptaresignificantlydifferent(P<0.05)
  • 5. Body Weight Gain (BWG) in starter period was observed in the lowest stocking density treatment (10 bird/m2 ) (p<0.05). Physical form of diet had no significant effect on feed intake, BW and Feed Conversion Ratio (FCR) throughout the periods. However the broiler feed intake was significantly influenced by stocking density and the decrease in the high stocking density group (18 bird/m2 ). Similarly, Mingbin et al. (2015) indicated that the use of pellet form in broiler diet had positive effects on body weight. However, the pellet form diet had no significant effect on other performance parameters (BWG, feed intake and FCR). These results were in agreement with Salari et al. (2006), who found that the form of diet and particle size had no significant effect on weight gain and dry matter intake. This observation suggested that high pellet quality may be necessary to fully obtain benefits of pelleting. Birds at normal stocking density (10 birds/m2 ) resulted in better body weight in starter period compared with the high stocking density (18 birds/m2 ). This indicates to a greater degree of stress on the performance. However, other performance parameters in other periods weren’t affected by different stocking density. Additionally, Buijs et al. (2009) detailed that at body weight 39 days of age was not significant between birds raised at various stocking densities (6, 15, 23, 33, 35, 41, 47 and 56 kg /m2 ). Interestingly, Houshmand et al. (2012) revealed that amid the growing stage (22-42 days) broilers raised at a high density had a inferior FCR contrasted and birds housed at normal density. As shown in Table 3, all carcass parameters (carcass, breast, thigh, back and neck), abdominal fat, some inner organ weight (small intestine, heart, liver, spleen and bursa of Fabricius) except for the gizzard percent weight were not significantly (p>0.05) influenced by the form of the diet and different stocking density. Use of mash diet significantly increased the gizzard percent weight (p<0.05). A similar observation was also recorded by Ravindran and Thomas (2004) who reported that carcass characteristics weren’t affected by stocking density. Previously, Ghorbani et al. (2012) Journal of Research in Biology (2016) 6(7): 2110-2116 2114 Reyhani et al., 2016 Treatment Breast Thigh Back Gizzard Small intestine Abdominal fat Heart Liver Spleen Bursa Feed form Mash 20.19 20.66 16.03 1.92a 1.78 1.45 2.02 2.31 0.1 0.11 Pellet 20.53 20.33 15.97 1.65b 1.89 1.48 0.53 2.24 0.1 0.11 SEM 0.3 0.1 0.3 0.04 0.03 0.07 1.00 0.03 0.03 0 Stock density 10 20.45 20.57 15.96 1.75 1.88 1.44 0.66 2.21 0.09 0.12 14 20.55 20.50 15.49 1.80 1.80 1.46 2.7 2.29 0.1 0.11 18 20.07 20.42 16.54 1.80 1.82 1.5 0.4 2.32 0.1 0.11 SEM 0.3 0.2 0.4 0.05 0.03 0.08 1.2 0.04 0.0 0.0 Interaction Mash 10 20.1 20.70 15.89 1.89 1.87 1.44 0.72 2.28 0.10 0.12 Pellet 10 20.8 20.44 16.04 1.61 1.89 1.43 0.60 2.15 0.09 0.11 Mash 14 20.93 21.04 15.65 2.01 1.71 1.38 4.96 2.36 0.09 0.11 Pellet 14 20.18 19.96 15.33 1.59 1.88 1.53 0.48 2.23 0.10 0.11 Mash 18 19.53 20.24 16.56 1.85 1.75 1.51 0.37 2.28 0.10 0.11 Pellet 18 20.60 20.59 16.53 1.75 1.89 1.49 0.52 2.35 0.10 0.11 SEM 0.54 0.29 0.61 0.07 0.05 0.12 1.76 0.06 0.005 0.001 a, b Means in column not sharing a common superscript are significantly different (p<0.05) Table 3. Effect of different feed form (mash and pellet) and stocking density on carcass characteristics and some inner organ weight (%)
  • 6. reported that the relative weight of gizzard increases by mash diet in compared with pellet form diet. The increase in relative weight of gizzard may enhance the digestive capacity of broilers. Mean antibody titers to sheep red blood cell, H: L ratio and litter moisture are presented in Table 4. Different physical feed type and stocking density had no significant effect on the antibody titers to sheep red blood and H/L ratio in the broiler chicks. However the litter moisture was significantly influenced by treatments (p<0.05). Use of pellet form diet and high stocking density treatment (18 bird/m2 ) significantly increased broiler litter moisture (P<0.05). The H:L proportion is a typical marker of stress in poultry (Heckert et al. 2002), and the blood leukocyte profile is impacted by stress. Decrease in the quantities of lymphocytes and monocytes and improvement in the quantities of heterophils, which prompts a higher H:L proportion, have been accounted for stressed animals (Stevenson and Taylor, 1988). Previously, Houshmand et al. (2012) reported that density had no significant effect on the heterophil: lymphocyte ratio too. In high stocking density, especially in hot and wet weather, litter moisture also gets increased and causes the increase in bacterial activity and ammonia production. Increased mortality can be explained by decreased animal welfare, such as bad air and litter quality, poor immune response, and poor feed intake. Litter quality has a large effect on dermatitis (Haslam et al., 2006). CONCLUSIONS Results showed that the use of pellet form diet and normal stocking density (10 bird/m2 ) can improve broiler body weight and litter quality. ACKNOWLEDGMENTS We sincerely thank the members of animal science department of Islamic Azad University, Varamin branch, Tehran, Iran. REFERENCES Banerjee GC. (1998). Poultry. 3rd edition, Oxford and IBH Publishing Co. Pvt. Ltd. Calcutta. 121p. Buijs S, Keeling L, Rettenbacher S, Van Poucke E and Tuyttens FAM. (2009). Stocking density effects on broiler welfare: identifying sensitive ranges for different Reyhani et al., 2016 2115 Journal of Research in Biology (2016) 6(7): 2110-2116 Treatment SRBC H/L (%) Litter moisture (%)S. No Feed form Mash 9.52 0.31 9.55b1 Pellet 9.47 0.31 12.12a2 SEM 0.5 0.00 0.53 Stock density4 10 10.12 0.318 8.66b 5 14 9.20 0.306 8.11b 6 18 9.16 0.310 15.74a 7 SEM 0.7 0.008 0.78 Interaction9 Mash 10 10.91 0.32 9.26b 10 Pellet 10 9.33 0.31 8.07b 11 Mash 14 9.08 0.30 7.91b 12 Pellet 14 9.33 0.30 8.32b 13 Mash 18 8.58 0.30 11.5b 14 Pellet 18 9.75 0.32 19.98a 15 SEM 1.03 0.01 0.916 Table 4. Effect of different feed form (mash and pellet) and stocking density on immunity and litter quality a, b Means in column not sharing a common superscript are significantly different (p<0.05)
  • 7. indicators. Poultry Science, 88(8): 1536-1543. Chewning CG, Stark CR and Brake J. (2012). Effects of particle size and feed form on broiler performance. The Journal of Applied Poultry Research, 21(4): 830–837. Dozier WA, Thaxton JP, Purswell JL, Olanrewaju HA, Branton SL and Roush WB. (2006). Stocking density effects on male broilers grown to 1.8 kilograms of body weight. Poultry Science, 85(2): 344–351. Dozier WA, Thaxton JP, Branton SL, Morgan GW, Miles DM, Roush WB, Lott BD and Vizzier-Thaxton Y. (2005). Stocking density effects on growth performance and processing yields of heavy broilers. Poultry Science, 84(8): 1332-1338. Eriflir M. (2002). Yerleflim sıklıı arttırılan bıldırcınların (Coturnix coturnix japonica) bazı biyokimyasal kan parametrelerindeki deoifliklikler. Turkish Journal of Veterinary Animal Science, 26: 491-496. Feddes JJ Emmanuel EJ and Zuidhof MJ. (2002). Broiler performance, body weight variance, feed and water intake, and carcass quality at different stocking densities. Poultry Science, 81(6): 774–779. Ghobadi Z and Karimi A. (2012). Effect of feed processing and enzyme supplementation of wheat-based diets on performance of broiler chicks. Journal of Applied Animal Research, 40(3): 260 266. Gross WB and Siegel HS. (1983). Evaluation of the heterophil/lymphocyte ratio as a measure of stress in chickens. Avian Disease, 27(4): 972-979. Haslam SM, Brown SN, Wilkins LJ Kestin SC, Warriss PD and Nicol CJ. (2006). Preliminary study to examine the utility of using foot burn or hock burn to assess aspects of housing conditions for broiler chicken. British Poultry Science, 47(1): 13–18. Heckert RA, Estevez I, Russek-Cohen E and Pettit-Riley R. (2002). Effects of density and perch availability on the immune status of broilers. Poultry Science, 81(4): 451–457. Houshmand M, Azhar K, Zulkifli I, Bejo MH and Kamyab A. (2012). Effects of prebiotic, protein level and stocking density on performance, immunity and stress indicators of broilers. Poultry Science, 91(2): 393-401. Jahan MS, Asaduzzaman M and Sarkar AK. (2006). Performance of broiler fed on mash, pellet and crumble. International Journal of Poultry Science, 5(3): 265–70. Mingbin Lv, Lei Yan, Zhengguo W, Sha An, Miaomiao Wu and Zunzhou Lv. (2015). Effects of feed form and feed particle size on growth performance, carcass characteristics and digestive tract development of broilers. Animal Nutrition, 1(3): 252–256. Ravindran V and Thomas DV. (2004). Performance and welfare of broilers as affected by stocking density and in-feed antibiotic supplementation. Proceedings of the 16th Australian Poultry Science Symposium, Sydney, New-South Wales, Australia, 9-11 ,135-138. Salari S, Kermanshahi H and Nasiri MH. (2006). Effect of sodium bentonite and comparison of pellet vs mash on performance of broiler chickens. International Journal of Poultry Science, 5(1): 31–4. SAS Institute. (2005). User’s Guide. Version 9. SAS Inst. Inc., Cary, NC. Stevenson JR and Taylor R. (1988). Effects of glucocorticoid and antiglucocorticoid hormones on leukocyte numbers and function. International Journal of Immunopharmacology, 10(1): 1–6. Journal of Research in Biology (2016) 6(7): 2110-2116 2116 Reyhani et al., 2016 Submit your articles online at www.jresearchbiology.com Advantages  Easy online submission  Complete Peer review  Affordable Charges  Quick processing  Extensive indexing  You retain your copyright submit@jresearchbiology.com www.jresearchbiology.com/Submit.php