CC BY
0JWPR
Journal of World's Poultry Research
2020, Scienceline Publication
J. World Poult. Res. 10(1): 36-40, March 25, 2020
Research Paper, PII: S2322455X2000005-10 License: CC BY 4.0
DOI: https://dx.doi.Org/10.36380/jwpr.2020.5
The Potency of Oral Administration of L-Citrulline as Anti Heat
Stress Agent in KUB Chicks
Edi Erwan1*, Triani Adelina1, Alaiddin Koto2 and Vebera Maslami1
1 Department of Animal Science, Faculty of Agriculture and Animal Science, State Islamic University of Sultan Syarif Kasim Riau, 28293, Indonesia 2Faculty of Syariah and Law, State Islamic University of Sultan Syarif Kasim Riau, 28293, Indonesia
Corresponding author's E-mail: erwan_edi@yahoo.com; ORCID: 0000-0002-5306-1582
ABSTRACT
Received: 28 Dec. 2019 Accepted: 04 Feb. 2020
L-citrulline (L-Cit) is considered one of the potential amino acids that are able to reduce body temperature in layer chicks. However, there are no reports on functions of L-Cit in KUB chicks. Therefore, the present study aimed to evaluate the effects of oral administration of L-Cit on feed intake, body temperature and plasma biochemical parameters of KUB chicks over two hours post-administration. Neonatal KUB chicks were orally administered with different doses of L-Cit (0, 3.75, 7.5 and 15 mmol/kg body weight). At 30, 60 and 120 min, feed intake was determined, also rectal temperature of chicks was measured by using a digital thermometer. At 120 min after administration, the blood samples were immediately collected through the jugular vein, then biochemical parameters of plasma (total cholesterol, triacylglycerol and total glucose) were analyzed. The obtained results indicated that feed intake, body temperature, and plasma metabolites were not significantly influenced by different doses of L-Cit. In conclusion, L-Cit could not able to improve feed intake, decrease body temperature and change plasma metabolites in KUB chicks.
Keywords: Body temperature, Feed intake, KUB chick, L- citrulline, Plasma metabolites
In general, native chickens play an important role in supporting income generation, especially in many underdeveloped countries (Padhi, 2016). KUB chickens are offspring of local chickens in several areas of West Java that are obtained after six generations of a systematically-controlled breeding system (Purbarani et al., 2019). The KUB chickens have several advantages compared to their previous generations as well as local chickens. For instance, KUB chickens have high rates of egg production (160-180 eggs/year), high hatchability, and low feed conversion ratio (Sartika, 2016). Besides that, KUB chicken is considered a meat type breed (Hidayah et al., 2019) and the high demand for chicken meat might potentially be fulfilled by increasing the productivity of KUB chicken.
In order to obtain good performance as an expression of the genetically improved breed, some efforts such as optimal management conditions (housing and feeding), is needed to increase biological and economic productivity. Many factors may influence poultry production in tropical and subtropical areas. The climatic environment is one of the important factors affect production efficiency in these
regions (Renaudeau et al., 2012). Poultry experiences heat distress when the combination of relative humidity and ambient temperature rise above the comfort zone (Teeter and Belay, 1996). There are concerns about reduced poultry production due to high ambient temperatures not only in tropical regions but also in some countries that occupy the temperate where heat stress is an occasional problem during the summer months.
Several nutritional strategies have been reported to overcome the adverse effects of heat stress in poultry (Teeter and Belay, 1996; Yahav, 2000; Erwan et al., 2013a, 2014, 2017; Chowdhury et al., 2015). Heat stress reduces laying percentage, egg size, eggshell durability, and body weight gain, as well as increases mortality in layers (Sterling et al., 2003; Lin et al., 2004; Franco-Jimenez and Beck, 2007). Also, it appears to have a detrimental impact on poultry meat quality, however, it has been reported that breast meat characteristics remain unaffected by heat stress (Goo et al., 2019).
Nutrients, especially amino acids, are widely used as anti-stress agents with regard to psychological and physiological stress (Yamane et al., 2009; Hamasu et al., 2010; Erwan et al., 2012). Amino acid supplements,
TBHiBS^HBaffiSa Erwan E, Adelina T, Koto A and Maslami V (2020). The Potency of Oral Administration of L-Citrullme as Anti Heat Stress Agent in KUB Chickens. J. World Poult. Res., 10 (1): 36-40. DOI: https://dx.doi.org/10.36380/jwpr.2020.5
particularly essential amino acids, has been used in attempts to overcome problems of heat stress in birds (Mendes et al., 1997; Daghir et al., 2003).
It has been reported that oral administration of L-citrulline (L-Cit) decreased the body temperature in layer chicks (Chowdhury et al., 2015). However, there are no reports on the function of L-Cit in KUB chicks. Therefore, the purpose of this study was to examine the effect of orally administered L- Cit on rectal temperature and some plasma biochemical parameters in KUB chicks.
MATERIALS AND METHODS
This research was conducted at the Poultry Division Field Laboratory, Faculty of Agriculture and Animal Science, State Islamic University of Sultan Syarif Kasim Riau, Indonesia in 2018.
Ethical approval
This experiment was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Animal, at the Faculty of Agriculture and Animal Science, State Islamic University of Sultan Syarif Kasim Riau, Pekanbaru, Indonesia
Animals
Thirty-two, one-day-old, KUB chicks were purchased from a local hatchery and housed in a wire-meshed cage (50 x 35 x 31 cm) at a constant temperature of 30 ± 1°C and with continuous light. Feed (Charoen Phokpand, Indonesia) and water were provided ad libitum. Feed composition is presented in table 1. One day before the experiment, chicks (4 days old) were weighed individually and assigned to treatment and control groups based on their body weight to produce uniform groups. The number of animals used in each group was kept to the minimum (n = 8) that would still ensure adequate statistical power.
Table 1. The percentage of nutrient content in ration
Nutrient Value
Crude Protein (%) 23.50
Crude Fiber (%) 1.88
Crude Fat (%) 5.87
Ca (%) 0.29
P (%) 0.15
ME (Kcal/kg) 3,050
Ca: Calcium, P: Phosphor, ME: Metabolizable Energy; *Commercial feed: 311-VIVO, PT, Charoen Pokphand, Indonesia; **Mineral Premix: Supplemented for kg of the diets: Vit. A, 12000 IU; D3, 2000 IU; E, 20 mg; K3, 3 mg; B2, 7 mg; B3, 12 mg; B5, 3 mg; B12, 0.03 mg; biotin, 0.1 mg; choline chloride, 300 mg; Mn, 130 mg; Fe, 70 mg; Zn, 60 mg; Cu,12 mg; I,1 mg; Se, 0.2 mg, and adequate antioxidant.
Experimental design
After a habituation period, chicks were randomly selected and divided into four groups each consisting of eight chicks. The chicks were reared individually in experimental cages and had ad libitum access to diet up to the time of the experiment. On the day of the experiment, each chick (5 days old) received L-Cit orally (treatment groups) or distilled water (control group) by the plastic needle on small syringe. Based on our recent report on Lor D-Asp and L-Cit in layer chicks (Erwan et al., 2013b, 2014; Chowdhury et al., 2015), treatment groups received 3.75, 7.5 or 15.0 mmol/ kg body weight as the low, medium and high dose, respectively. The chicks were fed ad libitum diets for 2 h immediately after the treatment. At the end of the experiments (2 h after L-Cit administration), birds were decapitated under chloroform anesthesia. Blood samples were collected in heparinized tubes and centrifuged for 15 min at 5,000 g, and the plasma was collected and stored at -20 °C until analysis took place.
Analysis of plasma biochemical parameters
The biochemical parameters of plasma including total glucose, total cholesterol, and triacylglycerol were determined with Microlab 300 (Vital Scientific, Netherland) as per the manufacturer's instructions. All the samples were assayed together and in a random sequence for each metabolite.
Measurement of feed intake and rectal temperature
At 30, 60 and 120 min after L-Cit administration, feed intake was calculated by measuring the reduction in the amount of feed consumed from a pre-weighed feeder. Similarly, rectal temperature of chicks was determined at 30, 60 and 120 min after L-Cit administration using a digital thermometer with an accuracy of ±0.1°C (Thermalert TH-5, Physitemp Instruments Inc., USA). It took about 5 sec to measure rectal temperature by inserting the probe.
Statistical analysis
Data on feed intake and rectal temperature were analyzed by repeated-measures two-way analysis of variance (ANOVA). Plasma metabolites were statistically analyzed by one-way ANOVA and regression equations. When significant differences were found, the comparison of means was performed using Tukey's test as a post hoc test. Differences were considered significant at p<0.05. Values are presented as means ± SEM. Statistical analysis
was carried out using the commercially available package StatView (Version 5, SAS Institute, Cary, USA). Data were checked by the Thompson rejection test to eliminate outliers (p<0.05), and the remaining data were used for the analysis among groups.
RESULTS AND DISCUSSION
The aim of the current study was to reveal whether the L-Cit amino acid influenced feed intake and have thermoregulatory functions in KUB chicks. Feed intake was not significantly affected by oral administration of several doses of L-Cit (data not shown). This result was consistent with the previous report by Erwan et al. (2016) who revealed that the inclusion of watermelon rind in ration during two weeks did not influence feed intake in broiler chicks. However, the aforementioned authors demonstrated that feed consumption increased when chicks fed for three weeks. The present findings also contrasted with previous findings in layers where L-Cit given for up to 9% caused a significant increase in feed intake in male layer chicks (Nguyen et al., 2019). These discrepancies in the feed intake in response to L-Cit may be due to variations in the experimental period, differences in doses, or species differences. Further experimentation would be needed to clarify the factors involved in this disparity.
As shown in diagram 1, rectal temperatures of KUB chicks did not change significantly by the oral administration of L-Cit (p >0.05). This result was not consistent with the previous report that indicated the oral administration of L-Cit decreased the body temperature in layer and broiler chicks (Erwan and Febriyanti, 2015; Chowdhury et al., 2017). The body temperature of KUB chicks ranged from 39.5 to 40.5 °C, which was still in the normal range. The different effects observed in this study can be attributed to the difference in chicken type. It is well known that indigenous chickens are more adaptable to stress than commercial chickens. Duangjinda (2017) revealed that indigenous chickens from a tropical environment had lower heterophil-to-lymphocyte ratio and mortality rate than those of commercial broilers under heat stress conditions. Similarly, Aengwanich (2007) revealed that Thai indigenous chickens and Thai crossbreed chickens can tolerate higher temperatures than broiler chickens.
Table 2 shows the effect of oral administration of different doses of L-Cit on certain biochemical parameters of plasma, indicating no significant changes were observed (p >0.05). The effect of oral administration of L-
Cit on total cholesterol, triacylglycerol, and glucose in the plasma of indigenous chicks is unclear. Watermelon rind contains a natural source of citrulline (Rimando and Perkins-Veazie, 2005). A previous study found that oral administration of red watermelon rind juice dose-dependently increased glucose and triacylglycerol in broiler chickens (Erwan et al., 2016). In contrast, Chowdhury et al. (2017) revealed that oral administration of L-Cit decreased glucose in layer male chicks. It is thought the differences in results may be due to different strains of chicken used.
Time (min)
Diagram 1. The effect of different doses of L-citrulline on body temperature of KUB chicks over two hours after oral administration.
Table 2. Effects of oral administration of different doses of L-citrulline on plasma metabolites in KUB chicks
Parameters L-citrulline (mmol)
0 3.75 7.5 15
Glucose (mg/d/) 237±20 229±14 261±8 226±18
Total cholesterol (mg/d/) 161±12 187±27 177±25 179±21
Triacylglycerol (mg/dl) 80±20 49±9 91±25 51±8
Values are expressed as mean ± SEM.
Heat stress affects plasma metabolites in poultry (Xie et al., 2015). An increase in plasma cholesterol indicates that chick has undergone heat stress (Olanrewaju et al., 2010). According to Altan et al. (2000), cholesterol in the body can come from de novo biosynthesis. De novo cholesterol biosynthesis is largely influenced by the broiler stress level (Setyadi et al., 2013). Blood cholesterol levels in this study ranged from 161 to 187 mg/dl, whereas Basmacioglu and Ergul (2005) reported that serum cholesterol contents of laying hens renged from 121to 142 mg/dl)
The results obtained in the present study indicated that triacylglycerol ranged from 49-91 mg/dl. When chickens are in a state of heat stress, plasma triacylglycerol levels drop (Sun et al., 2015). Environmental stress increases adrenocorticotropic hormone which stimulates the adrenal cortex to increase glucocorticoid secretion (von Borell and Ladewig, 1989). Blood glucose has a relationship with heat ambient temperature in poultry (Olanrewaju et al., 2010). Chowdhury et al. (2017) suggested that lower plasma glucose after oral administration of L-Cit may decrease the substrate for heat production and result in lower body temperature in chicks. Glucose in the blood serves as a buffer molecule for osmotic pressure thus normal osmotic pressure can be maintained even in a state of dehydration and heat stress (Guay et al., 2007) to maintain blood pressure and cardiovascular rhythms (Tan et al., 2010). In this study glucose concentration in chickens' blood ranged from 226-261 mg/dl. This range is included in the normal glucose levels, which indicates the chickens did not experience heat stress. Normal range glucose concentration in broiler blood is 230 - 370 mg/dl (Sulistyoningsih, 2004).
CONCLUSION
L-citrulline could not able to improve feed intake and affect on body temperature in KUB chickens. No significant changes in plasma glucose, total cholesterol and triacylglycerol were detected in the current study,indicating the oral administration of L-citrulline may not affect the metabolism of these components in KUB chicks.
DECLARATIONS
Acknowledgment
All authors are very grateful to the Dean of Faculty of Agriculture and Animal Science, State Islamic University of Sultan Syarif Kasim Riau, Indonesia for supporting.
Authors' contribution
Edi Erwan conducted the research, prepared data, performed the statistical analysis and wrote draft article. Vebera Maslami, Triani Adelina and Alaiddin Koto wrote the article. All authors checked and approved the final version of the manuscript.
Competing interests
The authors declare that they have no competing interests.
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