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0Performance, Serum Biochemical Parameters and Immunity in Broiler Chicks Fed Dietary Echinacea purpurea and
Thymus vulgaris Extracts
Hassan Habibi1* and Sobhan Firouzi2
1Assistant's professor of Agriculture and Natural Resources College, Persian Gulf University, Bushehr, Iran 2Avian Diseases Research Center, School of Veterinary Medicine, Shiraz University, Shiraz, Iran Corresponding author's Email: h.habibi@pgu.ac.ir
Received: 04 Aug 2017 Accepted: 06 Sept 2017
ABSTRACT
The objective of this study was to evaluate the effect of administrating herbal extracts of Echinacea purpurea and Thymus vulgaris into broilers drinking water on performance, immune response and serum biochemical and Phyto hemagglutinin. 270 day-old Ross chicks were assigned to nine dietary treatments in a randomized manner. Each treatment was given to two replicates of 15 birds. The variables of T. vulgaris extract were 1 % and 2% and variables of E. purpurea extract were 0%, 1% and 2% in drinking water. Body Weight (BW), Feed Intake (FI) and Feed Conversion Ratio (FCR) were recorded at the end of the experiment. Antibody responses against Newcastle disease viruses were measured after blood sampling at 42 days of age. The plant extracts did not affect BW, FI and FCR (P>0.05). Antibody titers against NDV were significantly affected by the administration of E. purpurea (P<0.05). The highest elevation was for the birds that were administrated with 2% E. purpurea from 1 to 42 days (P<0.05). Administration of thyme extracts had improved serum biochemical parameters as compared with Echinacea and control group. It was concluded that under these research conditions, high levels of E. purpurea extracts had increased the broiler chickens' immunity.
Key words: Broiler, Echinacea purpurea extract, Immunity, Performance, Thymus vulgaris extract.
JWPR
Journal of World's Poultry Research
2017, Scienceline Publication
J. World Poult. Res. 7(3): 123-128, Sept 25, 2017
Research Paper, PII: S2322455X1700015-7 License: CC BY 4.0
INTRODUCTION
Antibiotics have been widely used in poultry feed as growth promoters for more than 50 years. In the time that antibiotics were either used for curing or as growth promoters, some parts of the profitable microorganism will become damaged and resistance to diseases such as Salmonella and the other pathogens will decrease. Increased interest in curbing antibiotic use to reduce antimicrobial resistance has led to a growing interest in alternative growth promoters. Herbal extracts are being used as feed additives to improve performance, feed intake, secretion of digestive tract juices and immune system of animals especially under the intensive management systems (William and Losa, 2001; Amouzmehr et al., 2012).
The Echinacea purpurea is commonly known as an immune stimulating substance. Its palliative use in human medicine has been well established. E. purpurea has been shown to have non-specific immuno-stimulatory properties in vitro (Bauer and Wagner, 1991), including increased phagocytosis (Stotzem et al., 1992), increased cytokine production (Burger et al., 1997), and natural killer cell activity (See et al., 1997). Thymus vulgaris has received more attention due to its antioxidant (Bolukbasi and Erhan, 2007), antibacterial (Dorman and Deans, 2000; Ngouana Tadjong et al., 2017) anti-coccidial (Jamroz et al., 2003) and antifungal properties (Hertrampf, 2001). As, growth performance of animals is influenced strongly by the health and immune status, the objective of this study was to evaluate the effect of the utilization of Echinacea purpurea and Thymus vulgaris extracts in the feeding of
To cite this paper: Habibi H and Firouzi S (2017). Performance, Serum Biochemical Parameters and Immunity in Broiler Chicks Fed Dietary Echinacea purpurea and Thymus vulgaris Extracts. J. World Poult. Res., 7 (3): 123-128.
broilers on the growth performance and immune responses.
MATERIALS AND METHODS
270 day-old broiler chickens (Ross308) of mixed sex were randomly divided into nine groups (30 birds/group) and housed in pens of identical size in a deep litter system with wood shaving for flooring. Each group has 2 replicates (15 birds/pen). Strict sanitation practices were maintained in the house before and during the course of the experiment. All birds were fed a standard commercial diet based on corn and soybean meal and had free access to feed and water (Table 1). Treatment groups were:
1-Control group (plain water)
2- One mL/L (1%) of E. purpurea from 1 to 42 day
3-Two mL/L (2%) of E. purpurea from 1 to 42 day
4-One mL/L (1%) of E. purpurea from 21 to 42 day
5-Two mL/L (2%) of E. purpurea from 21 to 42 day
6-One mL/L (1%) of T. vulgaris from 1 to 42 day
7-Two mL/L (2%) of T. vulgaris from 1 to 42 day
8-One mL/L (1%) of T. vulgaris from 21 to 42 day
9-Two mL/L (2%) of T. vulgaris from 21 to 42 day
The live body weight of birds of all groups at 42 days of age was taken. Feed intake and their Feed Conversion Ratio (FCR) were calculated. Sera samples for subjecting to Haemagglutination Inhibition (HI) tests were obtained from all birds of each group following collection of 3mL blood (from wing vein) at 42 days of age to determine the antibody titers of Newcastle disease vaccines. Vaccination program is presented in table 2. To determine some of the
serum factors (HDL, LDL, TG, Cholesterol, Albumin, Total Protein, and Glucose), blood samples 42 days of age were collected. At 42 days of age, four birds per replicate were randomly chosen, slaughtered and their lymphoid organs (Bursa of Fabricius, spleen and Thymus) were collected, weighed and calculated as a percentage of live body weight. At d 42 of age, eight chickens per treatment were injected in the right wing with a Phytohemagglutinin (PHA) (Sigma-Aldrich, St. Louis, MO) solution (100 ^g in 100 ^l PBS (phosphate-buffered saline) and immediately in the left wing with 100 ^l PBS. The thickness of the right and left wing web was measured prior to the injection and at 6th, 12th, 24th, 48th, and 72nd hours post PHA injection using a digital caliper; then, the chickens were killed by cervical dislocation. The wing web swelling (WWS) response was expressed as a swelling index, calculated as follows: Swelling index = [(thickness of right wing web post PHA injection - initial thickness of right wing web) - (thickness of left wing web post PBS injection - initial thickness of left wing web)] (Konieczka et al., 2017). The data were analyzed using computerized statistical program (SPSS version 15.0) to determine the Mean ±SD of antibody titer and body weight. Significance differences were denoted by P < 0.05.
Ethical approval
This study was carried out in strict accordance with the recommendations of institutional guidelines for the care and use of laboratory animals. Chickens were humanly handled in respect of the ethical standards laid down in 1964 Declaration of Helsinki and its later amendments.
Table 1. Nutritional composition of the diet
Diet composition Starter (0-10d) Grower (11-28d) Finisher (29-42d)
Metabolizable energy, kcal/kg 2930 3050 3150
Crude protein % 23 19.5 18.5
Digestible lysine % 1.32 1.18 1
Methionine % 0.55 0.5 0.45
met + cys % 0.98 0.88 0.82
Threonine % 0.88 0.78 0.7
Tryptophan % 0.23 0.19 0.18
Arginine % 1.5 1.25 1.13
Calcium % 0.97 0.94 0.9
Available phosphorus % 0.48 0.45 0.42
Sodium % 0.16 0.16 0.16
Chlorine % 0.18 0.18 0.18
Table 2. Vaccination Program
Age Day 1 Day 8 Day 8 Day 16 Day 26
Vaccine Infectious Newcastle Newcastle- Infectious Bursal Infectious Bursal
Bronchitis-H120 Disease-B1 Influenza Disease-D78 Disease-D78
Route Corse spray Drinking water Injection Drinking water Drinking water
RESULTS
The effects of herbal extracts on chicken's body weight gain, total feed intake, and FCR have been presented in table 3. Results have shown that the administration of T. vulgaris from 21 to 42 days had affected growth performance more (body weight gain and feed consumption ratio) but these two parameters weren't statistically different (P>0.05). As shown in table 3, antibody titers against NDV were significantly affected by administration of E. purpurea (P<0.05). The highest elevation was for the birds that administrated 2% E. purpurea from 1 to 42 days (P<0.05).
Table 4 summarizes the data obtained on the effect of experimental treatments on serum hematological parameters. No significant influence of experimental diets on albumin was observed (P>0.05). The serum total protein, Triglyceride and HDL of the birds treated with (2%) of T. vulgaris from 1 to 42 days old were better than the others (P<0.05). The feeding of the broilers with (1%) of T. vulgaris from 1 to 42 days resulted in a marked (P>0.05) decrease in LDL- cholesterol concentration compared to other treatments. Hemoglobin concentration and Cholesterol in groups treated T. vulgaris from 1 to 42 days was respectively higher and lower than the other groups (P<0.05) respectively. Table 5 indicates the effect of treatments on lymphoid organs' weight at 42 day. As it is shown, the Bursa weight in birds treated with E. purpurea had increased and the differences in Spleen and Thymus weights were not statistically significant.
The effects of the dietary Echinacea purpurea and Thymus vulgaris extracts on PHA challenge on the WWS response are shown in Table 6. PHA challenge led to a higher increase in wing web thickness in chickens fed with E. purpurea diets than in those fed T. vulgaris diets (P<0.05). WWS was the highest at 24 h post PHA injection in group 3, post PHA injection (P<0.05).
DISCUSSION
There are no appetizing effects with both Echinacea and Thymus. The feed intake and FCR of the broilers didn't show a varying influence by administration with different dosages of Echinacea and Thymus in comparison with control group. Other authors have reported also a missing effect of various supplementations of mixed herbs on the feed intake of broilers and layers (Roth-Maier et al., 2005; Nasir and Grashorn, 2010). Toghyani et al. (2010) reported that the low dosage of Thyme has had a significant effect on broiler body weight and their feed conversion ratio. But Tekeli et al. (2006) and Demir et al. (2008) had reported opposite results; they found that thyme had no influence on broilers performance. Ngouana
Tadjong et al. (2017) reported that feed intake, live body weight, weight gain and feed conversion ratio are affected by using thyme and oregano essential oil in broiler diets.
Inability to improve feed conversion ratio with E. purpurea is in agreement with the findings of Habibian et al. (2011) who had also reported that E. purpurea supplementation as a feed additive didn't improvefeed conversion ratio. Absence of positive effect of some herbal extracts in some experiments may be due to using a smaller dose which was insufficient to produce its effect on poultry. Improving immunity in poultry production is very important to prevent common important diseases. There was a significant effect on the immunity of treated chicks in the present study. All chicks that were administrated E. purpurea had significantly higher antibody titer against T. vulgaris (P<0.05). Among them, the highest (8.9) increasing was belong to 2% E. purpurea extract from day old to 42 days (P<0.05).
Our findings indicated that the dietary E. purpurea could not only affect the PHA-induced swelling response, but also increased the weight of Bursa. The PHA-induced WWS is a good indicator of acquired immunity and allows the assessment of leukocyte interactions during the immune response (Konieczka et al., 2017). Increasing in antibody titer of the birds administrated E. purpurea are consonant with the increasing of the weight of bursa. The studies of the immune system have shown that some herbs such as coneflower (Echinacea purpurea) were most effective in achieving immune system improvement, because this herb had increased the stimulation of nonspecific immune system. It is though that immune enhancement of Echinacea is provided by certain polysaccharides, flavonoids and isobutylamides (Rehman et al., 1999). Also, herbs like thyme (Thymus vulgaris) that are rich in active compounds such as flavonoids extend the activity of vitamin C, act as antioxidants and may therefore enhance the immune function (Manach et al., 1996).
Obtained data from biochemical parameters showed that supplementing broiler diet with thyme extracts had insignificant better effect on cholesterol, triglyceride, LDL, total protein, albumin and hemoglobin ratio against supplementing Echinacea In agreement with our finding, Ali (2014) and Toghyani et al. (2010) reported supplementing diets with thyme leaves powder had no significant effect (P>0.05) on serum parameters. Thyme supplementation in broiler diets significantly increased glucose level compared to those of the control group. The possible reason for increasing serum glucose may be due to the abdominal lipids catabolism of gluconeogenesis process as feeding inclusion thyme by birds, since crushed thyme consumption in broiler chickens has been reported to increase the serum glucose as mentioned by El-Ghousein and Al-Beitawi (2009).
Table 3. Effects of E. purpurea and T. vulgaris on growth performance and antibody titer (log2 HI titer) against Newcastle virus (M ± SE)
Treatments Body weight (g) Feed intake (g) Feed conversion (g/g) Antibody titer (log2 HI titer)
Control 2477±226.1 4260 1.72 6.5a±0.15
E. p (1%) 1 to 42d 2412±268.3 4200 1.74 8.2b±.024
E. p (2%) 1 to 42d 2409±277.6 4170 1.73 8.9c±0.21
E. p (1%) 21 to 42d 2410±318.8 4170 1.73 7.4d±1
E. p (2%) 21 to 42d 2526±230 4320 1.71 8.1b±0.18
T. v (1%) 1 to 42d 2389±300.5 4325 1.81 6.5a±0.14
T. v (2%) 1 to 42d 2434±252.7 4260 1.75 6.6a±0.18
T. v (1%) 21 to 42d 2567±232.8 4340 1.69 6.4a ±0.13
T. v (2%) 21 to 42d 2460±272.1 4260 1.73 6.3a±0.19
The different superscripts on the same line are significantly different (P < 0.05); *Mean ± standard deviation
Table 4. Effect of experimental diets on serum biochemical parameters of broilers at d 42.
Factor(mg/dl) Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9
Blood Glucose 127±4.08ac 117.5±8.34a 123.5±7.18ac 129.25±10.5bc 116.5±4.79a 118.5±9.67a 122.75±9.97ac 131.25±12.97bc 124.5±10.5ac
Cholesterol 95.5±23.33ab 97.25±17.11ab 96±23.93ab 102.75±16.35b 95.5±19ab 72.75±7.5a 71.25±7.67a 85.25±11.58ab 84.25±11.35ab
Triglyceride 58±6.16b 53.50±9.84b 56±8.52b 51.25±12.12bc 54.50±4.65b 44.50±11.95ac 38.50±13.69a 41.25±19.5a 42.5±11.81a
LDL 47±11.9c 43.25±8.9bc 45.5±7.32c 46.75±6.39c 36.75±8.65abc 27.25±4.11a 31±9.09ab 30±9.2ab 29.25±9.28a
HDL 52.25±7.5a 61.25±11.5abc 60.25±4.92ab 64.75±4.11bc 59.75±8.05ab 72.50±4.79cd 76.25±5.90d 63.75±6.70bc 68±7.11bcd
Total protein 3.80±0.59a 3.97±0.22ab 4.09±0.11abc 4.22±0.11bc 3.83±0.09a 4.32±0.07bc 4.41±0.07c 4.18±0.06abc 4.05±0.19abc
Albumin 1.42±0.09a 1.47±0.03a 1.41±0.06a 1.39±0.04a 1.38±0.06a 1.46±0.03a 1.44±0.03a 1.39±0.1a 1.47±0.04a
Hemoglobin 7.85±0/32a 8.08±0.39ab 8.31±0.06bcd 7.97±0.24ab 8.25±0.09bc 9.40±0.11e 9.06±0.29e 8.49±0.3cd 8.69±0.16d
The different superscripts on the same line are significantly different (P < 0.05); *Mean ± standard deviation
Table 5. Effect of experimental diets on weights of lymphoid organs at day 42.
Lymphoid organ weight Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9
Bursa/BW (gr) 0.144±0.007a 0.168±0.005c 0.18±0.011d 0.177±0.011d 0.155±0.009abc 0.148±0.004ab 0.15±0.007ab 0.16±0.007bc 0.16±0.007bc
Spleen/BW (gr) 0.113±0.002abc 0.11±0.002abc 0.12±0.002d 0.114±0.002abc 0.118±0.005cd 0113±0.002abc 0.117±0.005abc 0.111±0.001a 0.113±0.002ab
Thymus/BW(gr) 0.195±0.03a 0.26±0.04b 0.247±0.03ab 0.24±0.05ab 0.242±0.03ab 0.212±0.03ab 0.212±0.04ab 0.22±0.04ab 0.227±0.04ab
The different superscripts on the same line are significantly different (P < 0.05); *Mean ± standard deviation
Table 6. Phytohemagglutinin (PHA) injection time on the wing web swelling response in chickens challenged with PHA
Wing web reaction (PHA-P) Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9
Stimulation index after 24 h (mm) 1.22±0.04ab 1.55±0.08f 1.76±0.1g 1.48±0.07f 1.52±0.06f 1.35±0.05cd 1.3±0.02bc 1.15±0.05 a 1.37±0.03cde
Stimulation index after 48 h (mm) 1.31±0.04bc 1.69±0.06g 1.71±0.05g 1.71±0.1g 1.47±0.04ef 1.36±0.04cd 1.53±0.04f 1.28±0.03b 1.44±0.01def
The different superscripts on the same line are significantly different (P < 0.05); *Mean ± standard deviation
CONCLUSION
Both herbal extracts (Echinacea and thyme) used in drinking water did not improve the performance of broilers. However, Echinacea extracts had significantly improved the immune response of birds to the NDV vaccine, therefore, more studies with different growth facility need to be performed to analyze the effect of herb extracts on the performance, carcass yield, hematology and immune response of poultry.
Competing interests
The authors have no competing interests to declare.
Authors' contributions
The present study was funded by H Habibi and S Firouzi. Habibi and Firouzi were also involved in the collection of data, statistical analysis and drafting of the manuscript. Both authors read and approved the final manuscript.
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