Herbs have acquired increasing interest as natural feed supplements and alternatives to antibiotics growth promoters in livestock production. The objectives of the current study to investigate the effects of supplementation by different levels of coriander seeds powder on growth performance indices and immune response of rats. A total of sixty albino rats were randomly assigned to four treatments with five replicates and fed for 30 days. The obtained results revealed that the final body weight, body gain, and body gain % in treatment groups fed diets supplemented with 3% and 5% coriander seeds powder were significantly higher as compared with the control group. Moreover, there were a significant improvement on feed utilization through feed intake and feed conversion ratio values obtained in treatment groups fed on 5%, 3%, and 2% coriander seeds powder in comparison with the control group. Interlukin-6 concentration level appeared significant improvement in rats fed 5% and 3% coriander seeds powder as compared with treatment group fed on 2% coriander seeds powder and the control group. From these result, it is concluded that coriander powder enhanced the growth performance and immune status of rats and can be used as natural feed supplements in their diets.
Using antibiotic growth promoters feed additives in the diets have played a key role in animal and poultry production. However, the vast majority for these antibiotics have been banned in large portions countries, especially those European Union, due to open wellbeing worry in regards to their residues in the animal products and the development of anti-microbial resistance (Schwarz et al., 2001; Lee et al., 2004). Currently, there is an increasing of interest to research on non-synthetic alternatives for antibiotics between the researchers. Phytogenic feed additives such as herbs and spices are commonly incorporated into the diets of livestock production to improve flavor and palatability, therefore enhancing feed efficiency utilization and growth performance (Windisch et al., 2008).
Herbs and spices are well known to exert potent antimicrobial properties in vitro against various pathogens, and as alternative feeding strategy to replace antibiotic growth promoters (Si et al.,2006; Lee et al.,2013) However, our information regarding their modes of action and aspects of their application animal diets is still set. Coriander seeds (Coriandrum sativum) are deemed both as an herb and as a spice. It has also been used as a medicine for thousands of years. As a medicinal plant, coriander has been used as an antifungal (Bas´ılico M. and Bas´ılico J. 1999), antioxidant (Chithra and Leelamma 1999; Helle et al., 2004), hypolipidemic (Chithra and Leelamma 2000), antimicrobial (Singh et al., 2002), hypocholesterolemic (Chithra and Leelamma 1997), and anticonvulsant substance (Hossein and Mohammad 2002). The major compounds present in Coriander seeds extract are linalool (67.70%); alpha-pinene (10.5%); gamma-terpinene (9.0%); geranyl acetate (4.0%); camphor (3.0%); and geraniol (1.9%) (Nadeem et al., 2013).
Coriandrum sativum L. (C. sativum) belongs to the family Apiaceae. Essential oil, flavonoids, fatty acids, and sterols have been isolated from different parts of C.sativum (Axel, 2008). The fruits (seed and pericarp) are the most widely used components of the coriander plant with the most important constituents being the essential oil and the fatty oil. The essential oil content is around 1% and the major component reported in the oil is linalool, in the range of 30–80% of total seed oil (Sahib et al., 2013). The phytochemical screening of Coriandrum sativum showed that it contained essential oil, tannins, reducing sugars, alkaloids, phenolics, flavonoids, fatty acids, sterols and glycosides. It also contained high nutritional values including proteins, oils, carbohydrates, fibers and wide range of minerals, trace elements and vitamins. The previous pharmacological studies manifested that it had antibacterial, antifungal, anthelmintic, insecticidal, antioxidant, cardiovascular, hypolipidemic, anti-inflammatory, analgesic, antidiabetic, anticancer, hepatoprotective, detoxification and many other pharmacological effects (Ali, 2016). Coriander, like many spices, contains antioxidants in both leaves and seeds, which can deferment the spoilage of food flavored with this spice (Goswami and Singhai, 2012). Supplementation of coriander seeds to rats fed with a high-fat diet appeared decreased levels of peroxides, free fatty acid and glutathione as well as increased activity of antioxidant enzymes (Chithra and Leelamma, 1999). The free radical scavenging efficiency of coriander oil has been partly attributed to the high contents of phospholipids available in coriander seed oil. Medication with polyphenolic portions of coriander seeds portions of coriander seeds adequately secured human lymphocytes from H2O2 induced oxidative stress and restored oxidative status to that of normal cells (Hashim et al., 2005). As stated by confirmation coriander possesses hepatoprotective efficiency against carbon tetrachloride (CCL4) intoxication, in vivo. (Pandey et al., 2011). The use of coriander as an anti-inflammatory factor is confirmed by a traditional formulation from Sri Lanka, Maharasnadhi Quather (MRQ), containing coriander seeds as one of its principal components. MRQ has been declared to have analgesic and anti-inflammatory properties both in animal models and human subjects. The formulation also increases pain bearing in rats by 57% after 1 h of treatment as assessed by the hot plate test (Thabrew et al., 2003). Causes and pathophysiology of inflammatory bowel disease (IBD) is still unknown and multifactorial (Sellin and Pasricha, 2006). Intestinal mucosal inflammation as a characteristic feature of IBD is induced by an increase in the activity of some mucosal immune cells where the T-helper cells play an important role (Sartor, 1997; Bouma and Strober, 2003). Various inflammatory mediators such as cyclooxygenases (COX-1 and COX-2), tumor necrosis factor-alpha (TNF-a), interleukin-6 (IL-6), and interleukin-12 (IL-12), the presence of highly activated inflammatory cells such as neutrophils, dendritic cells, macrophages, and excessive production of reactive oxygen species (ROS) have been implicated in the pathogenesis of the disease (Xavier and Podolsky, 2007).
Coriandrum sativum derivatives such as phenolic compounds and flavonoids show anti-inflammatory activity by controlling the levels of various inflammatory cytokines or inflammatory mediators including IL-1, IL-6, IL-10, TNF-α, NF-κB, NO, iNOS and COX-2 (Koretz and Rotblatt, 2004).The anti-inflammatory and anti-granuloma activities of Coriandrum sativum hydroalcoholic extract was studied in experimental models at the highest dose (32 mg/kg) where produced a significant reduction in paw edema and reduced serum IL-6 and IL-1 β levels in treated group as compared to control (Nair et al., 2013).
Furthermore, the mode of action of coriander seeds on interleukin 6 (IL-6) levels in rats has not been fully clarified yet. Therefore, the objective of the current study was accomplished to investigate the effects of using different levels of coriander seeds powder as feed supplements on growth performance parameters and IL-6 response as an immune marker in albino rats.
Materials and Methods
Animals and Experimental Design
The current experimental study was executed under the protocol approved by the college of applied medical sciences, Hail University, Kingdom of Saudi Arabia. Sixty albino rats weighting 118±0.60 gm were gained from the animal house in King Saud University, Riyadh, Saudi Arabia. The rats were randomly assigned to four treatments each treatment included five replicates with three rats per each. Experimental groups were assigned as follows-
Control = Basal diet without coriander seeds.
T1 = Basal diet plus 2% coriander seeds.
T2 = Basal diet plus 3% coriander seeds.
T3 = Basal diet plus 5% coriander seeds.
Rat Diets and Housing
Alfalfa meal, yellow corn, and sunflower seeds basal diets formulated to meet nutrients requirements of rats according to NRC, 1995 (Nutrient Requirements of Laboratory Animals), fourth revised edition. The ingredients and the composition of the basal formulated diets are shown in Table 1. Proximate analysis, which was conducted according to Official Methods of Analysis of AOAC International, 20th Edition (2016), showed no major deviation from calculated values. Different levels of coriander seeds were mixed with the rat basal diets according to the experimental design. Three albino rats were housed in each cage, which represent a replica, with wood shavings from a total 20 cages according to experimental design. The free access to feed and water was provided on an ad libitum basis. The rearing room was well ventilated and subjected to a photoperiod 12h light and 12h darkness throughout the experimental period (30 days).
Table 1: Formulated experimental diets used in this study
|Ingredients (gm/kg diet)||Treatments|
|Alfalfa Meal (17%)||160.0||160.0||150.0||170.0|
|Yellow corn, grain||398.0||398.0||390.0||370.0|
|Sunflower seeds meal||215.0||215.0||215.0||210.0|
|Dry Matter %||89.78||89.70||89.70||89.73|
|Crude protein %||15.67||15.67||15.67||15.74|
|Ether Extract %||3.31||3.58||3.74||4.0|
|Crude Fiber %||8.95||9.64||9.87||10.94|
1 Coriander seeds nutrients composition: Crude protein= 12.37 %, Ether Extract= 17.77%, Crude Fiber=41.9%, Energy= 2980 Kcal/kg, Dry Matter=88% (Source: USDA National Nutrient data base).
* Vit.&Min. Premix: Mineral (mg/kg diet): Calcium iodate- 1.50; Iron sulphate- 142.3; Cobalt carbonate- 0.43; Copper sulphate-4.30 mg; Manganous oxide-64.20; Zinc oxide-16.60;. Vitamin (/kg diet): Folate-2.53mg; Nicotinic acid-32.10mg; Ca-d-pantothenate-18.70mg; pyridoxine-1.57mg; Riboflavin-3.65mg; Vit B1- 10.50mg; Vit.A – 6,050.00 IU; Vit.D3-4,070.00 IU; Vit.K- 3.7mg; Vit E- 21.10mg; Choline – 615.00mg; d-Biotin-0.15mg; Vitamin B12- 0.005mg.
**Calculated according to feed stuffs ingredient analysis table 2014 edition.
Growth Performance Parameters
Body weight of rats were measured at the beginning and the end of experimental period and also the feed intake were recorded throughout the experimental period for each replica in treatment groups to estimate the growth performance parameters and feed intake per each using the following formulae: weight gain (WG; g/rat) =Wf-W0; weight gain percent=WG/W0*100; feed conversion ratio (FCR) = FI/ (Wf-W0), where Wf and W0were the final and first weights of the rats per group, respectively; and FI is feed intake.
Serum Samples Collection
Blood were collected from orbital venous plexus. Blood was centrifuged at 3000 g for 10 min using centrifuge and sera were separated by using dry Pasteur pipette. Labeled and stored in the refrigerator at -20 °C for analyses.
Measurement of Serum IL-6 Level
IL-6 levels were assayed in duplicate with all values expressed as a mean of two determinations. A standard curve was generated, using known concentrations of IL-6. The concentration of IL-6 in the samples and control serum samples was then determined from the standard curve. IL-6 was measured, using a validated commercial enzyme-linked immunosorbent assay (ELISA) (ENZO® life sciences). This assay uses a solid-phase anti–IL-6 antibody, bound to microtiters plates, that captures IL-6 in the sample assayed. Unbound protein is removed by washing, and anti–IL-6 antibody conjugated to horseradish peroxidase is added, with any excess conjugated antibody removed by further washing. The IL-6 bound is quantified by the addition of substrate solution (containing stabilized hydrogen peroxide and stabilized chromogen (tetramethylbenzidine). If antibody-peroxidase conjugate is present, the substrate is oxidized, and the intensity of the color that develops is proportional to the amount of bound IL-6. The reaction is stopped after 20 minutes of incubation at room temperature, and the color intensity is quantified by using a micro titer plate reader at a wavelength of 450 nm according to (Chard, 1990).
The experimental data were analyzed as a randomized block design where all data were subjected to one-way ANOVA by using the SPSS 22.00 statistical package. Duncan’s Multiple range test (Duncan, 1955) was done to evaluate any significant differences between means of treatment groups at the probability level of p<0.05. The model used was –
Yij = ? + Ai + eijk,
Where Yij = an observation,
μ = the common mean, Ai=the effect of the coriander seeds levels (i = 0, 2, 3 and 5%), and eijk=the effect of error.
Results and Discussion
Effect of Different Levels of Coriander Seeds Powder on Growth Performance Parameters
The final body weight, body weight gain, body gain percent, feed intake and feed conversion ration are present in Table 2. There were no significant differences in body weight of rats in the beginning of the experiment but there were significant differences in the final body weight between treatment groups where the higher significant values were obtained in the group fed on T3 followed by T2 then T1 in comparison to the control group (p <0.001).
Table 2: Effect of Different Levels of Coriander Seeds Powder on Growth Performance and Feed Conversion Ratio of Rats
|First Body Weight (g)||118.24||119.06||119.14||119.98||0.60||0.812|
|Final Body Weight (g)||183.68D||187.80C||193.52B||196.96A||1.27||<0.001|
|Body Gain (g)||65.44C||68.74BC||74.38AB||76.98A||1.36||0.002|
|Body Gain %||55.43B||57.85AB||62.48A||64.24A||1.27||0.037|
|Feed Intake (g)||98.56A||92.94B||83.92C||79.54C||1.84||<0.001|
|Feed Conversion Ratio||1.51A||1.36B||1.13C||1.04C||0.05||<0.001|
1 Values represent the means of 15 rats per each treatment group (5 rats/ replica).
T1 = 2% coriander seeds powder, T2 = 3% coriander seeds powder, T3 = 5% coriander seeds powder.
SEM- Standard error of Mean between treatment groups
ABCD Values in the same row with a different superscript differ significantly at p<0.05.
There were no significant differences in weight gain between T3 and T2; between T2 and T1 but there were significant differences due to the effect of treatment groups in comparison to the control group (p<0.05). There were significant increases in weight gain % in T3 and T2 than that in control group (p<0.05). Regarding the feed intake and feed conversion ratio (gm feed per gm gain), the dietary supplementation with coriander seeds powder in T3, T2, and T1 respectively significantly improved the feed intake and feed conversion ratio as compared the control group (P<0.001). The obtained results elucidated that dietary supplementation coriander up to 5% were effective in improving the growth performance parameters in albino rats. These findings were supported with that obtained by (Saeid and Al-Nasry 2010; Naeemasa et al., 2015) who showed that coriander seeds supplementation improved body weight (BW), body weight gain (BWG), and feed conversion ratio (FCR) in broiler chickens. The same results were shown by (Güler et al.,2005) who indicated that coriander seeds addition at a level of 2% improved BW and FCR in Japanese quails. This positive improvement could be referred to the essential oil, linalool, present in coriander seeds which have antimicrobial properties against harmful intestinal microorganisms and positive effect on body health and growth performance in poultry (Meena and Sethi, 1994; Elgayyar, 2001; Abuk et al., 2003). The improved FCR obtained with coriander treatments further confirmed the benefit of this herb. The improvement in feed conversion can also be attributed to the active component (linalool) found in coriander seeds, causing greater efficiency in the utilization of feed, resulting in enhanced growth. Additionally, (Przybilla and Weiss, 1998) reported that the mode of action of the herb mixtures on feed conversion is through the enhancement of the digestive functions. (Rajeshwari and Andallu, 2011; Windisch et al., 2008) reviewed that coriander essential oil is an excellent appetizer and helps in proper secretion of enzymes and digestive juices in the stomach, thus stimulating digestion and peristaltic motion which in turn improve FCR.
Effect of Coriander Seeds Powder on Serum IL-6 level
Obtained data revealed that, there were significant differences in serum interleukin-6 concentrations between the treatment groups (P˂0.001), where the highest value was observed in treatment group fed on 5% Coriander seeds powder supplementation followed by that fed on 3% Coriander seeds powder supplementation as compared with the control group, Meanwhile there was no significant difference between the treatment group fed on 2% Coriander seeds powder supplementation in comparison with the control group as shown in Table 3 and Figure 1.
The anti-inflammatory and analgesic effects of Coriandrum sativum seeds were evaluated in animal model. Meanwhile (Nair et al., 2013) evaluated the anti-inflammatory activity of Coriandrum sativum seeds (32 mg/kg) by using the subcutaneous cotton pellet implantation-induced granuloma formation and stimulation of peritoneal macrophages with complete Freund’s adjuvant. Serum tumor necrosis factor-α (TNF-α), IL-6, IL-1 β levels, and peritoneal macrophage expression of TNF-R1 were evaluated as markers of global inflammation. Coriandrum sativum at the highest dose (32 mg/kg) produced a significant reduction in Serum IL-6 and IL-1 β levels. Seeds of Coriandrum sativum possess both Anxiolytic and Central Analgesic activity (Pathan et al., 2011). The positive effect of coriander seeds powder on stimulation of IL-6 concentration as immune markers could be attributed to antioxidant activity (Helle et al., 2004) and active components present in coriander seeds extract such as linalool, alpha-pinene, gamma-terpinene, geranyl acetate, camphor, and geraniol which improve the health and immune status of the animals (Nadeem et al., 2013).
Table 3: Effect of Different Levels of Coriander Seeds Powder on Level of Serum IL6
1 Values represent the means of 15 rats per treatment group (5 rats/ replica).
T1 = 2% coriander seeds powder, T2 = 3% coriander seeds powder, T3 = 5% coriander seeds powder.
SEM: Standard error of Mean between treatment groups.
ABC Values in the same row with a different superscript differ significantly at p<0.05.
Fig 1: Effect of Different Levels of Coriander Seeds Powder on Serum IL6 Level
Under the situations of the current study, it could be concluded that dietary supplementation with coriander seeds powder up to 5% improved growth performance indices and immune status in albino rats. So up on these results dietary supplements with coriander seeds powder may be used as natural feed additives up to 5% without negative effects in animal diets.
Conflict of Interests
The authors declare that there is no conflict of interests regarding the publication of this paper.
This study was supported by College of Applied Medical Sciences, Hail University, Kingdom of Saudi Arabia.