Rev Mex Cienc Pecu 2017;8(1):11-21

http://dx.doi.org/10.22319/rmcp.v8i1.4309

Turmeric (Curcuma longa Linn.) as a phytogenic

growth promoter alternative for antibiotic and
comparable to mannan oligosaccharides for
broiler chicks

Youssef A. Attiaa*, Mohammed A. Al-Harthia, Saber S. Hassanb

ABSTRACT
This work aimed at investigating the potential as a growth enhancer of different dietary concentrations of turmeric
(Curcum a longa Linn.) as an alternative to oxytetracyline (OTC) as antibiotic and as comparable to mannan oligosaccharide for
broiler chicks. A total of 252 Hubbard broiler chicks at one day of age were distributed randomly in a straight run experimental
design among six treatments, each replicated seven times, with six unsexed chicks per replicate. The basal diet was
administered without supplements (control group) or supplemented with turmeric at 0.5, 1, and 2 g/kg diet, or with mannan
oligosacride (MOS) at 1 g/kg feed or with OTC at 50 mg/kg feed. Growth performance, carcass characteristics, meat quality
traits, blood biochemical constituents, antioxidant status and red blood cell (RBCs) were investigated. Turmeric
supplementation at 1 g/kg feed significantly improved feed conversion ratio (FCR) and European production index compared to
the control group and MOS groups. The results indicated that turmeric can be used at 1 kg/t feed as a phytogenic feed additive
as an alternative to OTC or MOS without negative effects on the productive and economic traits of broilers. There were no
differences from using OTC and MOS, while there was an increase in the European production efficiency index and the broilers’
health status.

KEY WORDS: Turmeric, Antibiotic, Prebiotics, Carcass traits, Meat quality, Blood biochemical, Blood biochemistry, Broiler.

RESUMEN
Este trabajo se realizó para investigar el potencial como promotor del crecimiento de diferentes concentraciones dietéticas
de cúrcuma (Curcum a longa Linn.) como una alternativa a los antibióticos y oxitetraciclina y comparable a oligosacáridos
mannan (MOS) para pollos de engorda. Un total de 252 pollos Hubbard de un día de edad se distribuyeron al azar en un diseño
experimental de seis tratamientos, siete repeticiones y con seis pollos sin sexar por repetición. La dieta basal se administró sin
suplementos (grupo control) o complementada con cúrcuma en dosis de 0.5, 1 y 2 g/kg de dieta, o con 1 g/kg de alimento de
oligosacárido mannan, o con oxitetraciclina 50 mg/kg de alimento. Se evaluó el crecimiento, características de la canal,
características de calidad de carne, constituyentes bioquímicos sanguíneos, estado antioxidante y glóbulos rojos. La
suplementación de cúrcuma en 1 g/kg alimento significativamente mejoró la tasa de conversión alimenticia y el Índice de
Producción Europea en comparación con el grupo control y grupo MOS. Los resultados indican que la cúrcuma puede usarse en
1 kg/t de alimento como una alternativa a oxitetraciclina o MOS sin efectos negativos sobre los rasgos productivos y económicos
de pollos de engorda. No hubo diferencias de uso entre oxitetraciclina y MOS, mientras que con cúrcuma hubo un aumento en
el índice de eficiencia de la producción europea y el estado de salud de los pollos.

PALABRAS CLAVE: Cúrcuma, Antibióticos, Probióticos, Calidad de canal, Calidad de carne, Química sanguínea, Pollos.

Received December 7, 2015. Accepted March 14, 2016.

a
Arid Land Agriculture Department, Faculty of Meteorology, Environment, and Arid Land Agriculture, King Abdulaziz University, Jeddah, Saudi Arabia. P.O. Box 80208, Jeddah
21589, Saudi Arabia Tel:+966568575961, fax:+9666952364.
b
Animal and Poultry Production Department, Faculty of Agriculture, Damanhour University, Egypt.

* Corresponding author: Youssef A. Attia. yaattia@kau.edu.sa.

Funding agent: This project was funded by the Deanship of Scientific Research (DSR), King Abdulaziz University, Jeddah, under grant No. (G-1436/155/205). The authors, therefore,
acknowledge with thanks DSR technical and financial support.

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 Youssef A. Attia, et al. / Rev Mex Cienc Pecu 2017;8(1):11-21

INTRODUCTION mode of action of MOS involves supplying

In the last few decades, antibiotic residuals intestinal microflora by nutrients (as prebiotics)
and the development of bacterial strains resistant and inhibiting the attachment of pathogenic
to antibiotics have become a problem(1) due to the bacteria, i.e. E. coli and Salmonella enteritidis, to
addition of antibiotics in animal feed the intestinal mucosa by binding the mannose
formulations(2,3,4). However, the prohibition of the receptors on the type 1 fimbriae(22). The positive
use of antibiotics in animal feed in Europe in 2006 effect of MOS on broiler performance was already
to improve the safety and security of the food reviewed(22).
chain caused significant health problems in Oxytetracyclines (OTC) are broad-spectrum
poultry, such as increasing the incidence of bacteriostatic agents derived from the
intestine necrotic enteritis and clostridia. These in bacteria Streptomyces. Oxytetracyclines prevent
turn caused major complications related to bacteria from multiplying while the host animal’s
decreasing animal welfare and increasing immune system deals with the original infection.
economic losses(4,5). Hence prebiotics, probiotics, The recommended dose is 5 to 50 g/t feed as a
synbiotics, herbs, spices and essential oils has continuous feed additive. In the literature,
been investigated as an alternative to antibiotics oxytetracyclines have been used for the
because of their antibacterial, antioxidant, amelioration of the growth of broilers, but the
digestive and metabolic enhancing effects(6).
Botanical compounds have been shown to be
potential alternatives to antibiotics for poultry Table 1. Fatty acid (FA) composition as percentage of
production(7,8,9). Turmeric is a member of the fatty acids and antioxidants indices of turmeric
Zingiberaceae family. It is mainly utilized in the
food industry to enhance the palatability, preservation As a % of As a % of
Fatty acid
FA1 FA2
and appearance of food. Turmeric contains different
bioactive compounds, such as curcumin, C6:0 0.166 ---
demethoxycurcumin, bisdemethoxycurcumin and C8:0 0.039 ---
tetrahydrocurcuminoids(10,11,12). These bioactive C10:0 0.347 ---
compounds have antioxidant, anti-inflammatory Undecanoic acid, C11:0 0.099 ---
and nematocidal activities(10,13,14,15), protective effects Tridecanoic acid, C13:0 0.282 ---
against mutagenicity and hepatocarcinogenicity Myristic acid, C14:0 0.94 ---
induced by aflatoxin(2,16) and against coccidiosis(5,17,18).
Pentadecenoic acid, C15:1 11.29 4.04
In the literature, the effects of turmeric
Palmtioleic C16:1 28.11 32.50
supplementation between 0 to 10 g/kg on chicken
Palmitic acid, C16:0 8..60 9.76
performance have been inconclusive. For example,
broilers fed diets supplemented with turmeric at 5 Heptadecenoic C 17:1 19.15 20.98
g/kg feed exhibited improved performance but did Linolenic C 18.3 --- 6.01
not affect serum total protein, albumin, globulin, Linoleic acid, C18:2c 15.70 18.63
ALKP, ALT and AST enzymes(1,6,19). On the other Oleic acid, C18:1 14.1 8.08
hand, turmeric powder did not significantly affect Stearic acid, C18:0 0.103 ---
growth performance or the carcass yield of broiler Arachidic acid, C20:0 0.16 ---
chickens(9,12,20). Turmeric powder at 0.6 and 0.9 SFA 10.73 9.76
g/kg alleviated the negative effect of aflatoxin
MUFA 72.65 61.66
B 1 on serum total protein, albumin and globulin,
boosted antioxidant defense enzymes, e.g. PUFA 15.70 24.64
catalase and superoxide dismutase, and UFA 89.35 86.30
decreased MDA (2). The levels of liver enzymes SFA/UFA ratio 0.120 0.113
(ALT and ALKP) were substantially reduced by Antioxidant activity inhibition, % 60.38 ---
feeding broilers turmeric powder at 5 g/kg(21).
Total phenolic compounds, mg/kg 14.25 ---
Prebiotics are also possible alternatives,
1 Represents the present sample of turmeric 2 according to Radwan(28), SFA=
particularly mannan oligosaccharides (MOS) Saturated fatty acids, MUFA= Mono unsaturated fatty acids, PUFA=
derived from Saccharomyces cerevisiae(22,23). The polyunsaturated fatty acids, UFA= Unsaturated fatty acids.

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 TURMERIC AS A PHYTOGENIC GROWTH PROMOTER

results have been contradictory(24,25). Thus this vegetable/plant feedstuffs according to Janssen(27).
research aims to investigate the growth promoting The fatty acids profile analysed according to
effect of turmeric (Curcuma longa Linn.) and to Radwan(28) (Table 1) after the extraction of
compare it to OTC and MOS on growth lipids(26). The total phenolic contents according to
performance, carcass characteristics, meat quality, Balinsky et al(29) and the antioxidant activity (%)
serum biochemical constituents and health status inhibition, determined to Benzie et al
during the 1st through 35th days of age of broiler methodology(30).
chickens.

Chicks, diets and experimental design

MATERIAL AND METHODS A total of 252 Hubbard broiler chicks 1 d of
age were used in this experiment. They were fed
Source of turmeric, fatty acid profiles and the experimental diets (Table 2) according to a
antioxidant indices
two-phase feeding system, with a starter-grower
Turmeric purchased from the local market in diet from d 1 to 27 and a finisher diet from d 28 to
a powder form was used in this experiment. The 35. The chicks were distributed randomly in a
chemical analyses of the experimental diets were complete randomized design among six
according to AOAC(26), meanwhile, metabolizable treatments. Each was replicated seven times with
energy value was calculated using the equation for six unsexed chicks per replicate. The basal diet was

Table 2. Diets composition and nutrient profiles of the experimental diets percentage as fed basis
Starter-grower diets 1-27 d of age (kg/t) Finisher diets 28-35 d of age (kg/t)
Ingredients/ profiles
0 0.5 1.0 2.0 0 0.5 1.0 2.0
Maize 508 507.5 507 506 600 599.5 599 598
Full fat soybean 80 80 80 80 100 100 100 100
Soybean meal 335 335 335 335 230 230 230 230
Vegetable oil blend 40 40 40 40 37 37 37 37
Turmeric powder 0.0 0.5 1.0 2.0 0.00 0.5 1.0 2.0
Sodium chloride 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0
Dl- methionine 1.5 1.5 1.5 1.5 1.0 1.0 1.0 1.0
L-lysine 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0
Dicalcium phosphate 18.5 18.5 18.5 18.5 15.0 15.0 15.0 15.0
Limestone 10 10 10 10 10 10 10 10
Vit. + Min. mixture1 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0
Total 1000 1000 1000 1000 1000 1000 1000 1000

Calculated2 and chemical composition3 (g/kg)

ME MJ/kg diet2 .1297 .1297 .1297 .1297 .1346 .1346 .1346 .1346
Dry matter3 897 895 898 896 895 897 898 891
Crude protein3 222 223 220 221 189 187 189 187
Calcium2 9.2 9.2 9.2 9.2 8.2 8.2 8.2 8.2
Inorganic phosphorus2 4.9 4.9 4.9 4.9 4.1 4.1 4.1 4.1
Methionine+cystine2 8.5 8.5 8.5 8.5 7.4 7.4 7.4 7.4
Lysine2 13.0 13.0 13.0 13.0 11.0 11.0 11.0 11.0
Crude fibre3 52.3 52.3 52.3 52.3 56.5 54.7 55.5 57.5
Crude fat3 88.3 88.3 88.3 88.3 89.1 87.1 88.1 89.3
Ash3 85.3 84.3 83.7 86.4 91.2 90.8 88.5 89.0
1Vit+Min mix. provides per kilogram of the diet: Vit. A, 12000 IU, vit. E (dl-α-tocopheryl acetate) 20 mg, menadione 2.3 mg, Vit. D3, 2200 ICU, riboflavin 5.5 mg,
calcium pantothenate 12 mg, nicotinic acid 50 mg, Choline 250 mg, vit. B12 10 µg, vit. B6 3 mg, thiamine 3 mg, folic acid 1 mg, d-biotin 0.05 mg. Trace mineral
(mg/ kg of diet): Mn 80 Zn 60, Fe 35, Cu 8, and Selenium 0.1 mg.

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 Youssef A. Attia, et al. / Rev Mex Cienc Pecu 2017;8(1):11-21

formulated to be isocaloric and isonitrogenous and At d 35 of age, seven blood samples were
to meet the nutrient requirements(31). The basal collected in both not-heparinized and heparinized
diet was administered without the tested tubes from each treatment represented all
supplements (the control group) or supplemented treatment replicates. The serum was separated by
with turmeric at 0.5 (T_0.5), 1 (T_1), and 2 (T_2) centrifugation at 1,500 xɡ for 10 min at 4 °C and
g/kg diet. The concentrations of turmeric stored at -18 °C until analysis. The selected serum
supplementation was chosen based in previous biochemical profile such as serum total protein and
studies(12,20,21) in which 0 to 10 g/kg was included albumin concentrations (g/dL), alanine amino-
in broiler diets with inconclusive results. The basal transferase (ALT) and aspartate aminotransferase
diet was also supplemented with mannan (AST), (µ/L), alkaline phosphatase (ALKP) enzymes,
oligosaccharides (MOS; Alltech Inc., Nicholasville, total antioxidant capacity (TAC) as an indicator of
KY, USA) at 1 g/kg diet or oxytetracycline (OTC) at antioxidant status, and malnodialdehyde (MDA as
50 mg/kg diet. OTC is a broad-spectrum a biomarker for lipid peroxidation respectively
bacteriostatic agent derived from the bacteria were determined using commercial diagnostic kits
Streptomyces. The recommended dose is 5 to 50 (Diamond Diagnostics Company, Cairo,
g/t feed as a continuous feed additive. Terramycin Egypt)(32,33). Globulin concentration (g/100 mL)
(OTC) is a registered trademark of Pfizer, Inc., was calculated as the difference between total
USA. It is US FDA NADA (new animal drug protein and albumin. Red blood cell (RBCs)
application) #95-143, approved by the FDA characteristics, including haemoglobin (Hgb),
7870000 101-9010-07 and licensed to Phibro packed cell volume (PCV), mean corpuscular
Animal Health Corporation for OTC HCl. haemoglobin (MCH), mean corpuscular volume
(MCV) and mean corpuscular haemoglobin
Broilers husbandry concentration (MCHC), were measured as
previously cited(32,33). Haemagglutination (HINDV)
Chicks were kept in battery brooders
inhibition for New Castle disease virus was
(40×45×60 cm) under similar managerial and
determined according to Snyder et al(34).
hygienic conditions in semi-opened housing. Water
and mash feeds were offered ad libitum. The Upon necropsy, the intestine was removed,
brooding temperature was 34, 32 and 30 ºC during thoroughly washed with a physiological saline
the 1st, 2nd and 3rd wk of age, respectively. During (0.9% NaCl) solution, blotted on filter paper and
21 to 35 d of age, the average ambient then buffered with formalin 10%. The fixed
temperature and relative humidity (RH) were 30 ± specimens were processed using a conventional
3 °C to 45 ± 4 %, respectively. The light-dark cycle paraffin embedding technique. From the prepared
was 23:1. paraffin blocks, 5 mm thick sections were obtained
and stained with haematoxylin and eosin for light
Data collection microscopic examination(35). In order to determine
the length of the villi, 5 villi were measured on each
Body weight was recorded at the 1st, 14th, 27th segment for all groups. The villi lengths were
and 35th d of age; body weight gain, feed intake measured from their base upwards to the end of
and the feed conversion ratio (FCR) were the villus. The morphometric measurements were
calculated for the periods 1-14, 1-27, and 1-35 d taken in a binocular microscope equipped with a
of age. At 35 d of age, seven chickens from each clear Nikon camera and coupled with an image-
treatment representing all replicates were analysing system from Optika(36).
randomly taken and slaughtered to determine their
carcass characteristics. In addition, the lymphoid
organs, including the thymus, spleen and bursa of Statistical analysis
Fabricius, were removed and weighed. Meat Data were analysed using the SAS software
quality traits (n= 7 samples/treatment) program(37), using a completely randomized
represented all replicates, such as chemical design, considering the replicate as the
composition (dry matter, protein, lipid and ash) experimental unit according to the following
and physical characteristics (pH, colour of meat, model:
water holding capacity [WHC] and tenderness)
were carried out as previously reported(32). Yi, j=µ+Тi+εi(j)

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 TURMERIC AS A PHYTOGENIC GROWTH PROMOTER

With Yi,j being any observation for which X1=i results, as displayed in Table 1, describe the fatty
(i and j denote the level of the factor and the acids content of turmeric, antioxidant activity
replication within the level of the factor, inhibition and total phenolic compounds. The
respectively); μ= general location parameter; Ti= results indicate that linoleic acid is the dominant
is the effect of having treatment level I; εi(j)= is polyunsaturated fatty acid and palmitoleic acid is
the random error. the dominant monounsaturated fatty acid. These
indicate that turmeric is a good source of
Mean differences were tested by the Tukey’s
unsaturated fatty acids. The antioxidant activity
studentized test(37) using P≤0.05; although when
inhibition and total phenolic compounds are 60.38
P value was great than 0.05 and less than 10 was
% and 14.25 mg/g, revealing a potential
reported as trend. Before analysis, all percentages
antioxidant activity.
were converted to arc sin to normalize data
distribution.
Growth performance
Data for broiler performance are shown in
RESULTS Table 3. The results showed that different
supplements did not significantly affect BWG of
Chemical composition, fatty acids, chickens during different experiment period except
antioxidant activity percentage inhibition for a trend for greater (P≤0.089) growth of T_0.5,
and total phenolic compounds
and the MOS groups during d 15-27 and T_0.5 and
Chemical composition of turmeric showed T_1 (P≤0.095) during the whole experimental
89.7 % dry matter, 5.8 % crude protein, 4.7 % period (d 1-35 of age) in comparison to the control
ether extract, 4.2 % ash 3.5 % crude fiber and group, T_2 group and OTC groups and the control
71.5 % nitrogen free extract and calculated ME group, T_2 group, MOS and the OTC groups,
value was found to be 3,664 kcal/kg turmeric. The respectively.

Table 3. Growth performance of broiler chickens fed diets supplemented with different concentrations of turmeric,
mannanoligosacchride and oxytetracycline
Curcumin g/ kg diet
Criteria Control MOS OTC SEM P-value
0.5 1 2
Body weight and body weight gain, g
Body weight at 1 d 46 46 48 47 45 46 1.01 0.524
BWG 1-14 d of age 386 352 376 341 377 347 13.9 0.139
BWG 15-27 d of age 837 954 882 829 905 836 34.1 0.089
BWG 28-35 d of age 495 527 580 565 510 599 40.1 .0401
BWG 1-35 d of age 1718 1833 1838 1735 1792 1781 34.2 0.095
Feed intake, g/chick/ period
1-14 d of age 646 a 607ab 560ab 511b 649a 549ab 23.6 0.001
15-27 d of age 1495 abc 1652 a 1465 abc 1416 bc 1597ab 1316c 52.8 0.001
28-35 d of age 994 1093 1072 1002 1045 1252 77.6 0.225
1-35 d of age 3135ab 3352a 3097ab 2929b 3292a 3118ab 73.3 0.005
Feed conversion ratio, kg feed/kg gain
1-14 d of age 1.67ab 1.72a 1.49c 1.50c 1.72a 1.58bc 0.022 0.001
15-27 d of age 1.78 a 1.74 ab 1.66 ab 1.71 ab 1.77a 1.59b 0.038 0.012
28-35 d of age 2.01 2.09 1.85 1.74 2.04 2.19 0.109 0.076
1-35 d of age 1.82a 1.83a 1.68b 1.69b 1.84a 1.75ab 0.021 0.001
Survival rate and European production efficiency index (EPEI)
Survival rate, % 100 100 100 100 100 100 0 ND
EPEI 262b 279ab 303a 286ab 271b 282ab 5.98 0.001
MOS= Mannoligaosacchride; OTC=Oxytiteracycline; BWG= Body weight gain; SEM= Standard error of means.
a,b,c Differences among means within a column within each factor not sharing similar superscripts are significant (P<0.05),

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 Youssef A. Attia, et al. / Rev Mex Cienc Pecu 2017;8(1):11-21

Feed intake during the different experimental most of the experimental groups except for the T-
periods was significantly affected by the different 0.5, T_1 and T_2 groups. During d 28-35 of age,
treatments. During d 1-14 of age, the T_2 group there was a trend for improved FCR of groups on
consumed significantly less feed than the control T_1 and T_2 in comparison to the other
and MOS groups. During d 15-27 of age, the T_2 experimental groups. For the whole period, the
and OTC groups reduced feed intake in comparison T_1 and T_2 groups significantly boosted FCR in
to the T_0.5 group. In addition, OTC groups comparison to the other groups except for OTC.
reduced feed intake in comparison to the MOS
The survival rate was 100 % in the different
group. For the 1-35 d period, the T_2 group
experiment groups. The European Production
significantly decreased feed intake in comparison
Efficiency Index of the T_1 group was significantly
to the T_0.5 and MOS groups.
higher than that of the control and MOS groups.
During most of the experimental periods, the Other groups exhibited intermediate values.
different supplements significantly affected FCR.
During d 1-14 of age, the T_1, T_2 and OTC
Carcasses characteristics and inner body
groups significantly improved FCR in comparison
organs
to the other groups, but the T_1 and T_2 groups
had more favorable effects than the OTC group as The results for carcass traits, relative weight
the difference between OTC and control group was of internal organs, chemical composition and
not significant. During d 15-27 of age, the OTC physical parameters of meat are presented in
group significantly improved FCR in comparison to Table 4. Most of the traits were significantly

Table 4. Carcass characteristics, inner body organs and meat quality of broiler chickens fed diets supplemented with different
concentrations of turmeric, mannanoligosacchride and oxytetracycline
Curcumin g/kg diet
Criteria Control MOS OTC SEM P value
0.5 1 2
Carcass characteristics and inner body organs
Dressing,% 72.7a 68.1b 69.4ab 69.9ab 68.7ab 66.6b 0.951 0.006
Abdominal fat,% 0.759 0.715 0.670 0.634 0.544 0.673 0.091 0.654
Proventiculus,% 0.582a 0.495ab 0.379b 0.532ab 0.604a 0.505ab 0.056 0.010
Gizzard,% 1.38a 1.32a 0.889b 1.25a 1.11ab 1.15ab 0.079 0.005
Liver,% 2.36 2.23 2.16 2.25 2.34 2.48 0.111 0.422
Heart,% 0.540a 0.352b 0.430ab 0.533a 0.484ab 0.450ab 0.038 0.022
Pancreas,% 0.283 0.274 0.214 0.253 0.299 0.228 0.019 0.039
Intestinal,% 7.93 8.53 8.13 8.97 7.65 7.99 0.451 0.332
Intestinal villi length, μm 1261c 1435b 1406b 1289b 1255c 1806a 57.4 0.001
Spleen weight, % 0.102ab 0.084b 0.114ab 0.091b 0.162a 0.088b 0.014 0.008
Thymus weight, % 0.394a 0.377ab 0.335b 0.481a 0.307b 0.340b 0.078 0.004
Fabricius bursa weight, % 0.134ab 0.117 ab 0.179 a 0.157 ab 0.159ab 0.100b 0.017 0.040
Chemical composition of meat,%
Dry matter 25.1 24.9 25.0 25.0 24.9 24.9 0.064 0.215
Crude protein 19.2 19.1 19.3 19.0 18.9 19.1 0.101 0.086
Lipids 4.72 4.75 4.58 4.73 4.81 4.64 0.071 0.294
Ash 0.973 0.993 0.990 0.987 0.983 0.990 0.012 0.879
Physical characteristics of meat
pH 5.93 6.08 6.01 6.00 6.02 5.98 0.079 0.862
Color, optical density 0.197 0.210 0.212 0.203 0.209 0.209 0.094 0.864
Tenderness cm2/g 9.82 10.02 10.19 10.26 9.86 10.26 0.066 0.579
WHC, cm2/g 17.48 18.25 17.48 17.92 17.82 17.98 0.059 0.315
SEM= Standard error of means, MOS= Mannoligaosacchride, OTC=Oxytiteracycline, pH= hydrogen power; WHC= Water holding capacity.
a,b,c Differences among means within a column within each factor not sharing similar superscripts are significant (P<0.05).

16
 TURMERIC AS A PHYTOGENIC GROWTH PROMOTER

affected by the treatments with the exception of T_0.5 group. The Fabricius bursa of the T_1 group
the relative weight of the abdominal fat, liver, was significantly higher than that of only the OTC
pancreas (although F value was significant group.
P≤0.039 for only pancreas) and intestine. Dressing
percentage significantly decreased due to T_0.5
and OTC supplements in comparison to the control Meat quality
group. It was found that the T_1 group significantly Table 4 shows the content of the dry matter,
decreased proventriculus in comparison to the protein, lipids and ash of the meat, as well as the
control and MOS groups, and also decreased rate of physical traits such as pH, color, WHC and
gizzard in comparison with the control, T_0.5 and tenderness. These traits were not significantly
T_2 groups. The heart percentage was significantly affected by the different supplementations, but
lower in the T_0.5 group than in the control and T_2 there was a trend (P≤0.086) for higher CP of T_1
groups. Most supplemented groups, except the MOS group than that of MOS group.
group, significantly increased intestinal villi length in
comparison to the control one, with the OTC group
displaying the greatest effect. Blood biochemical, liver leakage, antibody
titer and red blood cells characteristics
Lymphoid organs such as the spleen, thymus
and Fabricius bursa were significantly affected by The blood serum biochemical components are
the dietary supplementations. The MOS group shown in Table 5. Different supplements
exhibited significantly greater spleen weight than significantly affected serum total protein, globulin,
the T_0.5, T_2 and OTC groups. The thymus ALKP, ALT, AST, AST/ALT ratio, HI NDV and TAC.
percentage was greater of the control and T_2 It was found that the T_0.5, MOS and OTC groups
groups than those of the other groups except for had a significant increase in the total protein in

Table 5. Serum biochemical, liver leakage markers, antioxidant indices and red blood cell parameters of broiler chickens fed
diets supplemented with different concentrations of turmeric, mannanoligosacchride and oxytetracycline
Curcumin g/kg diet
Criteria Control MOS OTC SEM P value
0.5 1 2
Serum protein metabolites
Total protein, g/dL 6.18ab 6.30a 5.73c 5.93bc 6.40a 6.40a 0.066 0.001
Albumin, g/dL 3.30 3.27 3.12 3.12 3.07 3.07 0.103 0.628
Globulin, g/dL 2.88 3.02 2.60 2.80 3.33 3.32 0.143 0.032
Albumin to globulin ratio 1.16 1.083 1.24 1.14 0.928 0.942 0.087 0.051
Liver leakage and antibody markers
Alkaline phosphatase, U/L 9.25c 10.75bc 12.75a 11.25ab 11.75ab 11.00abc 0.431 0.008
ALT, U/L 63.0a 62.0a 62.3a 61.8a 60.8a 58.8b 0.512 0.001
AST,U/L 54.3a 50.0b 52.3ab 52.0ab 52.3ab 53.3ab 0.731 0.017
AST/ALT ratio 0.862ab 0.806b 0.840b 0.843b 0.863ab 0.909a 0.014 0.003
HINDV, Log2 4.37b 5.48ab 6.73a 5.62ab 5.15ab 4.12b 0.325 0.013
Antioxidant indices
TAC, mmol/l 416b 439a 434a 422b 431a 431a 1.43 0.001
MDA, µmol/l 10.8 11.3 11.0 11.3 10.8 11.3 0.471 0.920
Red blood cell parameters
RBCs, (106/mm3) 1.58 1.70 1.67 1.62 1.72 1.70 0.039 0.071
Hgb, g/dL 11.8a 10.8ab 11.8a 10.8ab 10.3b 11.3ab 0.271 0.004
PCV,% 32.0ab 32.2ab 33.2a 32.8a 31.2b 32.2ab 0.381 0.008
MCV, µm3/RBC 205a 190ab 200a 200a 181b 190ab 4.65 0.006
MCH, pg 74.4a 63.0cd 70.2ab 65.4bcd 60.0d 67.0bc 1.61 0.001
MCHC,% 36.2a 32.8b 34.8ab 32.6b 32.8b 34.8ab 0.721 0.002
MOS= Mannoligaosacchride; OTC=Oxytiteracycline; SEM= Standard error of means; ALT= Alanine amino transferase; AST= Aspartate amino transferase;
HINDV=haemagglutination inhibition for new castle disease virus; TAC= Total antioxidant capacity; MAD= Malnodialdehyde; Hgb=Hemoglobin; PCV= Packed
cell volume; MCV= Mean cell volume; MCH= Mean cell hemoglobin; MCHC= Mean cell hemoglobin concentration.
a,b,c Differences among means within a column within each factor not sharing similar superscripts are significant (P<0.05).

17
 Youssef A. Attia, et al. / Rev Mex Cienc Pecu 2017;8(1):11-21

comparison to the other groups except for the improved the growth of chickens exposed to
control group. The latter group had also greater aflatoxins(40,41), and alleviated the negative
total protein than T_1 group. Differences in serum influences of Eimeria infection(18,19,42) and of heat
globulin were not significant among different stress(39). These potential effects of turmeric could
means of groups. ALKP was significantly higher for be attributed to its curcuminoids (3 to 5 %, as
the groups supplemented with T_1, T_2 and MOS found in turmeric powder), bisdemethoxy
in comparison to the control group; the T_1 group curcumin and demethoxy curcumin, the principle
showed the greatest effect and T_0.5 and OTC active compounds in turmeric(43). These
exhibited intermediate values. compounds show a wide spectrum of biological
activities including antioxidant, antibacterial,
The antibiotic supplemented group had
antifungal, antiprotozoal, antiviral, anticoccidial
significantly decreased ALT in comparison to the
and anti-inflammatory properties, digestion- and
other groups, while the T_0.5 group had significantly
absorption-enhancing effects, and protection
decreased AST in comparison to only the control
effects against coccidiosis and toxins(5,19,44).
group. In addition, the AST/ALT ratio of the turmeric
Turmeric also improves liver and bile functions
groups was significantly lower than that of the OTC
through increased bile secretions, protects the
group. HINDV was the highest of T_1 group while
stomach from ulcers and reduces liver toxins.
the lowest was from the control and OTC groups.
These improvements can enhance digestion,
Most of supplemented groups except that T_2 group
metabolic processes and nutrient utilisation for
had significantly higher TACs than that of the control
growth through stimulation of protein synthesis by
group, with the T_0.5 group exhibiting the
the chicken enzymatic system(6,11). Turmeric has
greatest TAC. There were no significant
been observed to enhance the intestinal lipases,
differences in MAD among the different groups.
amylase, trypsin and chymotrypsin secretions(45).
The different treatments had a significant This is similar to our findings regarding the
effect on most of the hematological traits except increase in the length and width of villi in the
for RBCs (P≤0.071). Hgb, PCV, MCV, MCH and intestinal, which are also similar to other
MCHC were the lowest in the MOS group, while findings(45). Therefore, the improvement in the
Hgb, PCV, MCV of the T_1 group had the highest growth performance due to turmeric
values. MCH and MCHC were the highest in the supplementation to broilers’ diets can be partly
control group but did not significantly differ from attributed to improving the ecology and function of
the T_1 group. the digestive tract of chickens. On the other hand,
turmeric did not show constant effects on growth
performance as had been reported(21). No
DISCUSSION significant positive effect of turmeric powder at
The present results indicate that turmeric is a between 3.03 and 10 g/kg diet on the growth
potential source of nutrients, poly-unsaturated performance of broiler chickens(9,12). This
fatty acids and antioxidants(38), and T_1 improved inconsistency in the reviewed results can be
growth performance and the European production attributed to the different qualities of feed, breeder
efficiency index. This indicates that 1 g/kg turmeric and age of the broilers, statistical design, doses of
is adequate as an alternative growth promoter that turmeric and the sanitary and environmental
could replace OTC and have a better impact on conditions. The improved production index by
productive performance than MOS for both FCR 15.6 % due to inclusion of turmeric powder at
and the European production index. 1 g/kg is in the range cited in the literature of
1.5 % (39) and 11.8 %(6) when turmeric was
The potential effect of turmeric on growth supplemented at 5 g/kg feed.
performance and the production index of broilers
are in line with those reported elsewhere(11,20,39). Turmeric, particularly at 1 g/kg feed, induced
The aforementioned authors concluded that adaptive changes in the different body organs. The
turmeric supplementation at the rate of 1 to 10 decreased proventriculus and gizzard and
g/kg improved growth performance of broiler increased intestinal villi length indicated enhanced
chickens without adverse effects on mortality. In digestive function that can explain the increased
addition, turmeric supplementation at 5 g/kg feed performance, meat protein and somewhat

18
 TURMERIC AS A PHYTOGENIC GROWTH PROMOTER

decrease in meat lipid of the broilers on the T_1 longa did not affect crude protein or extracts of
treatment. On the other hand, turmeric at 1 and 2 breast and thigh meat as well as organoleptic tests
g/kg diet had no negative effects on carcass traits. (smell, flavour, colour and tenderness).
Meat quality is an important concept in broiler
Lymphoid organs, antibody level, antioxidant
production nowadays and improved postharvest
status and blood metabolites are a good markers
quality and shelf life is essential(32). The present
of health status of the animal. The impact of
findings indicate that turmeric is a beneficial feed
turmeric concentrations on lymphoid organs
additive due to phytochemicals, such as curcumin,
indicates that different concentrations of turmeric
AR-turmerone, methylcurcumin and other active
did not affect spleen, thymus and Fabricius bursa
compounds that could improve carcass quality and
percentages, but MOS increased percent spleen
reduce spoilage(1,5). This increase in the quality of
and decreased thymus and did not significantly
the carcass traits of broilers could be attributed to
affect Fabricius bursa and 28-d HINDV titer in
its antimicrobial effect, which improves the shelf
comparison with the control and antibiotic groups.
life of the carcasses(1,6,11).
In the literature, the inclusion of turmeric powder
Despite the absence of a significant effect of increased the spleen weight and did not affect the
turmeric in this study on the relative weight of Bursa and thymus weight index(11), spleen and
abdominal fat, liver and intestines, there was a bursa of Fabricius(9,12) and relative weight of the
numerical decease in percentage abdominal fat of spleen, bursa or thymus(20). These results reveal
11.7 % and 16.5 % and in liver of 8.5 and 4.7 % that turmeric is a safe phytogenic feed supplement
due to turmeric supplementation at 1 and 2 g/kg, for chickens and may enhance their immune
respectively. The positive effect of turmeric in response as measured by specific antibody titres,
abdominal fat and liver could be attributed to its as reviewed by others(5).
negative influence on liver fatty acid synthesis as The changes in serum metabolites indicate
manifested by an increase in meat CP and the that, except for the decrease in serum total
decrease in meat lipid of T_1 group. In literature, protein, turmeric supplementation at different
liver triacylglycerol and plasma triacylglycerol in doses did not affect serum albumin, globulin and
the VLDL fraction and liver cholesterol significantly albumen to globulin and indices of hepatocellular
decreased, but the activity of hepatic acyl-CoA leakage (ALT, AST and AST/ALT). There were,
oxidase increased(9,46). In addition, turmeric at a rate however, numerical decreases in the AST and
of 3 g/kg feed reduced the meat fat content and AST/ALT ratio, which show a potential decease in
increased the carcass quality of broilers(11,47,48). hepatocellular leakage markers that could be
In partial agreement with the present results, attributed to the significant increase in TAC
turmeric supplementation at 5 g/kg feed did not (antioxidants index) of broilers fed turmeric
significantly affect percent dressing, liver, gizzard supplemented-diets. Similarly, serum ALT and AST
and heart, but significantly increased the were not affected by turmeric powder(49). In
proventriculus(6). In addition, turmeric at the same addition, turmeric powder at 0.6 and 0.9
dose significantly increased percent dressing, weight alleviated the negative effect of aflatoxin B1 on
of the breast and thigh, but did not affect percent serum total protein, albumin and globulin,
liver, heart and gizzard(6). In other studies, turmeric boosted antioxidant defence enzymes, e.g.
supplementation did not significantly affect the catalase and superoxide dismutase, and
weight of the carcass, heart, pancreas or intestine(9), decreased MDA (2). The levels of liver enzymes
gall bladder(11), the ready-to-cook carcass, liver, (ALT and ALKP) were substantially reduced by
pancreas, heart, gizzard, proventriculus, abdominal feeding broilers turmeric powder(22). On the other
fat and length of the entire small intestinal, hand, turmeric at 5 g/kg feed did not affect serum
duodenum, jejunum and ileum(12). However, total protein, albumin, globulin, ALKP, ALT and AST
turmeric decreased the abdominal fat and liver enzymes(20).
percentages(9,11) and increased percentage of the The increase in the Hgb and PCV of broilers
entire small intestinal and the ileum weight(12). supplemented with turmeric at 1 g/kg feed
The effect of turmeric, MOS and OTC on meat indicates an improvement in health status. This
quality are in partial agreement with those can be attributed to the antioxidant capacity of
reported elsewhere(11), who showed that curcuma turmeric and its digestive-enhancing effect that

19
 Youssef A. Attia, et al. / Rev Mex Cienc Pecu 2017;8(1):11-21

may improve iron absorption. Similar results were 3. Attia YA, Zeweil HS, Alsaffar AA, El-Shafy AS. Effect of non-
antibiotic feed additives as an alternative to flavomycin on broiler
reported by for RBCs(11) and for PCV(12). Also, found chickens production. Archiv Geflügelk 2011;75:40-48.
that turmeric improved the health status of 4. Attia YA, Al-Harthi MA. Nigella seed oil as an alternative to
broilers(20). In accordance with the present results, antibiotic growth promoters for broiler Chickens. Europ Poult Sci
no mortality up to 35 d of age in broilers was 2015;79:DOI, 10.1399/eps.2015.80.
observed when turmeric was supplemented at 0.5 % 5. Eevuri TR, Putturu R. Use of certain herbal preparations in broiler
in the broiler diet(6), and less mortality was feeds - A review, Vet World 2013;6:172-179.

observed at 0.1 % inclusion(39). 6. Durrani FR, Ismail M, Sultan A, Suhail SM, Chand N, Durrani
Z. Effect of different levels of feed added turmeric Curcuma
It was found that turmeric at 1 g/kg feed had longa on the performance of broiler chicks. J Agric Biol Sci
2006;1:9-11.
effects comparable to MOS and OTC on growth,
dressing percentage and lymphoid organs, and 7. Nasir Z, Grashorn MA. Echinacea, A potential feed and water
additive in poultry and swine production. Arch Geflügelk
was better by 8.7 % than MOS for FCR. The lower 2009;73:227-236.
feed utilization of the MOS group might have been
8. Nasir Z, Grashorn MA. Effects of Echinacea purpurea and Nigella
due to their lowest villi length. OTC induced the sativa supplementation on broiler performance, carcass and
highest increase in intestinal villi, with no meat quality. J Anim Feed Sci 2010;19:94-104.
difference in FCR and production index from the 9. Nouzarian R, Tabeidian SA, Toghyani M, Ghalamkari G, Toghyani
turmeric groups. The effect of OTC seems to be M. Effect of turmeric powder on performance, carcass traits,
humoral immune responses, and serum metabolites in broiler
related to the anti-inflammatory action of OTC chickens. J Anim Feed Sci 2011;20:389-400.
rather than to its antibiotic effect(26,50,51). On the
10. Kiuchi F, Goto Y, Sugimoto N, Akao N, Kondo K, Tsuda Y.
other hand, MOS and OTC increased serum total Nematocidal activity of turmeric, Synergistic action of
protein compared to the intermediate and highest curcuminoids. Chem Pharm Bull 1 9 9 3 ; 41:1640-1643.
turmeric doses. These results indicated that 11. Al-Sultan SI. The effect of Curcuma longa turmeric on overall
turmeric had antibiotic-like effects due to its performance of broiler chickens. Int J Poult Sci 2003;2:351-353.
antimicrobial and anti-inflammatory effects. 12. Sadeghi GH, Karimi A, Padidar JSH, Azizi T, Daneshmand A.
Effects of cinnamon, thyme and turmeric infusions on the
performance and immune response in of 1-to 21-day-old male
broilers. Braz J Poult Sci 2012;14:15-20.
CONCLUSION AND IMPLICATIONS
13. Osawa T, Sugiyama Y, Inayoshi M, Kawakisi S. Anti-oxidative
Turmeric can be used at 1 kg/t feed as a activity of tetrahydrocurcuminoids. Biotech Biochem
phytogenic feed additive as an alternative to OTC 1995;59:1609-1616.
or MOS without negative effects on the productive 14. Wuthi-Udomler M, Grisanapan W, Luanratana O, Caichompoo W.
and economic traits of broilers. There were no Anti-fungal activities of plant extracts. South East Asian J Trop
Med Public Health 2000;31:178-182.
differences from using OTC and MOS, while there
was an increase in the European production 15. Alia BH, Marrif H, Noureldayemc SA, Bakheitd AO, Blunden G.
Biological properties of curcumin, a review. NPC 2006;1:509-
efficiency index and the broilers’ health status. 521.

16. Soni, KB, Lahiri M, Chackradeo P, Bhide SV, Kuttan R. Protective

effect of food additives on aflatoxin-induced mutagenicity and
ACKNOWLEDGMENTS hepatocarcinogenicity. Cancer Letters 1997;115:129-133.

This project was funded by the Deanship of 17. Allen PC, Fetterer RH. Recent advances in biology and
immunobiology of Eimeria species and in diagnosis and control
Scientific Research (DSR) at King Abdulaziz of infection with these coccidian parasites of poultry. Clin
University, Jeddah, under grant G-1436/155/205. Microbiol Rev 2002;15:58-65.
The authors, therefore, acknowledge with thanks 18. Lee SH, Lillehoj HS, Jang SI, Kim DK, Ionescu C, Bravo D. Effect
DSR for technical and financial support. of dietary curcuma, capsicum, and lentinus on enhancing local
immunity against Eimeria acervulina infection. J Poult Sci
2010;47:89-95.

LITERATURE CITED 19. Abou-Elkhair R, Ahmed HA, S. Selim S. Effects of black pepper
(Piper nigrum), turmeric powder (Curcuma Longa) and coriander
1. Windisch WM, Schedle K, Plitzner C, Kroismayr A. Use of seeds (Coriandrum sativum) and their combinations as feed
phytogenic products as feed additives for swine and poultry. J additives on growth performance, carcass traits, some blood
Anim Sci 2008;86:140-148. parameters and humoral immune response of broiler chickens.
Asian-Austral J Anim Sci 2014;27:847-854.
2. Ahmadi F. Effect of Turmeric Curcumin longa powder on
performance, oxidative stress state and some of blood 20. Mehala C, Moorthy M. Production performance of broilers fed
parameters in broilers fed on diets containing aflatoxin. Global with aloe vera and Curcuma longa (turmeric). Int J Poult Sci
Vet 2 0 1 0 ; 5:312-317. 2008;7:852-856.

20
 TURMERIC AS A PHYTOGENIC GROWTH PROMOTER

21. Emadi M, Kermanshahi H. Effect of turmeric rhizome powder on 36. Aptekmann K, Artoni S, Stefanini M, Orsi M. Morphometric
the activity of some blood enzymes in broiler chickens. Int J Poult analysis of the intestine of domestic quails (Coturnix coturnix
Sci 2007;6:48-51. japonica) treated with different levels of dietary calcium Anat
Histol Embryol 2001;30:277-280.
22. Hooge DM. Meat-analysis of broiler chicken pen trials evaluating
dietary mannan oligosaccharide 1993-2003. Int J Poult Sci 37. SAS, Institute. User’s guide. Ver 9.2, 2nd ed. SAS institute Inc.
2004;3:163-174. Cary NC. USA. 2009.

23. Attia YA, Abd El Hamid EA, Ismaiel AM, El Nagar ASh. The 38. Gur S, Turgat-Bubk D, Gur N. Antimicrobial activates and some
detoxication of nitrate by two antioxidants or a probiotic and the fatty acids of turmeric, ginger root and linseed used in the
effects on blood and seminal plasma profiles and reproductive treatment of infectious diseases. World J Agric Sci 2006;2:439-
function of NZW rabbit bucks. Anim 2013;7:591-601. 442.

24. Ogunwole OA, Abu OA, Adepoju IA. Performance and carcass 39. Kumar M, Choudhary RS, Vaishnav JK. Effect of supplemental
characteristics of broiler finishers fed acidifier based diets. Pak J prebiotic, probiotic and turmeric in diet on the performance of
Nut 2011;10:631-636. broiler chicks during summer. Ind J Poult Sci 2005;40:137-141.

25. Khadem A, Soler L, Everaert N, Niewold TA. Growth promotion 40. Yarru LP, Settivari RS, Gowda NKS, Antoniou E, Ledoux DR,
in broilers by both oxytetracycline and Macleaya cordata extract Rottinghaus GE. Effects of turmeric Curcuma longa on the
is based on their anti-inflammatory properties. Br J Nut expression of hepatic genes associated with biotransformation,
2014;112:1110-1118. antioxidant, and immune systems in broiler chicks fed aflatoxin.
Poult Sci 2009;88:2620-2627.
26. Official methods of analysis of AOAC International. Official
Methods of Analysis 16th ed. 2007. 41. Gowda NKS, Ledoux DR, Rottinghaus GE, Bermudez AJ, Chen YC.
Efficacy of turmeric Curcuma longa, containing a known level of
27. Janssen WM. European table of energy values for poultry curcumin, and a hydrated sodium- calcium aluminosilicate to
feedstuffs, 3rd ed. Wageningen, The Netherlands. 1989. ameliorate the adverse effects of aflatoxin in broiler chicks. Poult
28. Radwan SS. Coupling of two dimension thin layer Sci 2008;87:1125-1130.
chromatography with gas chromatography for the quantitative 42. Abbas RZ, Iqbal Z, Khan MN, Zafar MA, Zia, MA. Anticoccidial
analysis of lipids glasses and their constituent fatty acids. J activity of Curcuma longa L. in broilers. Braz Arch Biol
Chromatography Sci 1978;16:538-542. Technol 2010;53:63-67.
29. Blainski A, Lopes GC, De Mello JCP. Application and analysis of 43. Roughley PJ, Whiting DA. Experiments in the biosynthesis of
the folin ciocalteu method for the determination of the total curcumin. J Chem Soc Perkin Trans 1973;1:2379-2388.
phenolic content from Limonium brasiliense L. Molecules
2013;18:6852-6865. 44. Masuda T, Maekawa T, Hidaka K, Bando H, Takeda Y,
Yamaguchi H. Chemical studies on antioxidant mechanism of
30. Benzie IF, Strain JJ. Ferric reducing/antioxidant power assay, curcumin, analysis of oxidative coupling products from curcumin
direct measure of total antioxidant activity of biological fluids and and linoleate. J Agric Food Chem 2001;49:2539-2547.
modified version for simultaneous measurement of total
antioxidant power and ascorbic acid concentration. Meth 45. Rajput N, Muhammah N, Yan R, Zhong X, Wang T. Effect of
Enzymol 1999;299:15-27. dietary supplementation of curcumin on growth performance,
intestinal morphology and nutrients utilization of broiler chicks.
31. National Research Council, NRC. Nutrient Requirements of
Poultry. 9th ed. Washington DC: National Academic Press; 1994. J Poult Sci 2012;50:44-52.

32. Attia YA, El-Tahawy WS, Abd El-Hamid AE, Nizza A, El-Kelway 46. Asai A, Miyazawa T. Dietary curcuminoid prevent high-fat
MI, Al-Harthi MA, Bovera F. Effect of feed form, pellet diameter induced lipid accumulation in the rate liver epididymal adipose
and enzymes supplementation on carcass characteristics, meat tissue. J Nutr 2001;131:2932-2935.
quality, blood plasma constituents and stress indicators of
47. Samarasinghe K, Wenk C, Silva KSFT, Gunasekera JMDM.
broilers. Archiv Tierzucht 2014;57:1-14.
Turmeric Curcuma longa root powder and mannanoligosaccha-
33. Attia YA, Abd Al-Hamid AE, Ibrahim MS, Al-Harthi MA, Bovera F, rides as alternatives to antibiotics in broiler chicken diets. Asian-
El-Naggar A Sh. Productive performance, biochemical and Aust J Anim Sci 2003;16:1495-1500.
hematological traits of broiler chicks supplemented with propolis,
bee pollen, and mannan oligosaccharides continuously or 48. Emadi M, Kermanshahi H. Effect of turmeric rhizome powder on
intermittently. Livestock Sci 2014;164:87-95. performance and carcass characteristics of broiler chickens. Int J
Poult Sci 2006;5:1069-1072.
34. Snyder DB, Marquardt WW, Mallinson ET, Savage PK, Allen DC.
Rapid serological profiling by enzyme-linked immunosorbent 49. Reddy T. Effect of herbal preparations on the promoters. Poultry
assay. III. Simultaneous measurements of antibody titers to fortune performance of broilers [M.V.Sc., thesis]. Sri
infectious bronchitis, infectious bursal disease, and Newcastle Venkateswara Veterinary University, Tirupathi. India. 2010.
disease viruses in a single serum dilution. Avian Disease 50. Dibner JJ, Richards JD. Antibiotic growth promoters in agriculture,
1984;28:12-24. history and mode of action. Poult Sci 2005;84:634-643.
35. Bancroft JD, Layton C, Suvarna SK. Bancroft’s theory and practice 51. Niewold TA. The non-antibiotic anti-inflammatory effect of
of histological techniques. 7th ed, Churchill Livingstone: Elsevier; antimicrobial growth promoters, the real mode of action? A
2013. hypothesis. Poult Sci 2007;86:605-609.

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