The effects of nucleotide supplementation on the productivity, immune response and meat quality of broiler chicken reared under different environmental conditions

Mohammed Salah^1,2, Edjeng Suprijatna², M Luthfi Djauhari² and Y B I Vitus Dwi²

¹ Faculty of Animal Production, University of Khartoum, Department of Poultry Production, Sudan
² Faculty of Animal and Agricultural Sciences, Diponegoro University, Indonesia
m.salah1900@gmail.com

Abstract

The purposes of this experiment were to study the effects of dietary nucleotide supplementation on the broiler productivity, immune response and meat quality in broilers reared under different environmental conditions. This experiment was designed as factorial 3 x 3, with a total of 135 unsexed commercial chicks at fourteen-day of age, divided into three different environmental conditions; hot environment (32±1°C), comfortable environment (23 ±1°C), and natural environment, and fed with three different levels of nucleotide supplementation; 0, 0.5 and 1 g/kg of feed up to 35-days age. The feed intake, body weight gain (BWG), FCR, haemagglutination inhibition (HI) and immune organs weight were measured. The breast muscle was used to examine the pH, color, WHC, cooking loss, and Fe content. The results revealed that nucleotide improved body weight gain and FCR, but did not affected the feed intake. Hot environment reduced feed intake and body weight gain compared with natural and comfortable environments. Antibody levels against Newcastle and bursa of fabricius weight were improved by nucleotide supplement. The supplementation of dietary nucleotide increased iron content and had no effects on color, WHC, cooking loss and pH of the carcass. High ambient temperature reduced the pH but had no effect on other meat quality parameters. In conclusion, the present study clearly demonstrated that nucleotides improved broiler performance and immune response under heat stress.

Key words: productivity, immune, meat quality, different environment and broiler chicken

Introduction

The tropical and subtropical regions are very hot where temperature can rise above 30°C. Poultry are sensitive to high environmental temperature, with an optimum temperature for performance of 19 to 22°C for laying chickens and 18 to 22°C for developing broilers (Charles and Walker 2002). Poultry producers in these countries suffer from the high impact of heat on birds, and because of the high costs of the closed house system, which most of them cannot afford. They are therefore looking for alternatives that reduce the bad effects of heat stress such as feed additives.

Additives such as probiotics, prebiotics and enzymes may favor proper utilization of feed and safeguard gut integrity of chickens leading to better absorption of nutrients (Prakash et al 2016).

Nucleotides (as asbiotechnology product) are one of the potential feed additives, which can play positive roles in broiler productivity in the tropical and subtropical areas. Nucleotides, for years, were not considered essential nutrients for use in any dietary programs. It was thought that all organisms can supply sufficient amounts of nucleotides to meet their physiological demands via de novo synthesis and a salvage pathway (Andrino and Valeriano 2012).

However, during times of extraordinary stress, such asthe periods of rapid growth, certain disease states,reproduction, environmental change, recovery from injury, limited nutrient intake or disturbed endogenous synthesis of nucleotides, their availability could limit the maturation of fast dividing tissues with a low biosynthetic capacity, such as the intestine and immune cells. Thus, trillions of additional nucleotides must be readily available for cell proliferation (Esteve-Garcia et al 2007; Chiofalo et al 2011).

There are a number of previous studies that used nucleotides under different conditions of stress such as high stocking density combined with dirty litter in broiler houses (Jung and Batal 2012), chronic diarrhea in rats (Nunez et al 1990) andoxidative stress in pigs (Sauer et al 2011). There are no studies on the effect of nucleotides on environmental stress, so the present investigation aimed to study the effects of dietary nucleotide supplementation on productivity, immune response and meat quality in broilers reared under different ambient temperatures.

Material and methods

Experimental Site

The study was conducted at the Instructional Poultry Farm and laboratories of Faculty of Animal and Agriculture Science, Diponegoro University, Semarang. The house was divided into three parts, each part provided different environmental temperature. The first part of the house had been supplied with a source of heat and digital thermostats to achieve the hot environment with ambient temperature 32±1°C. The second part provided with cooling source to get the comfortable environment (23±1°C). Open system was used in the third part of the house, which depends on the surrounding natural temperature.

Source of Nucleotide

The nucleotide used in the experiment was BioNutrend® nucleotide Feed Grade. produced by CBH Co. Ltd., China. The product contained adenosine, guanosine, cytidine and uridine 5'-monophosphates (5'-AMP, 5'-GMP, 5'-CMP, and 5'-GMP), extracted from yeasts (Saccharomyces cerevisiae) through enzymatic hydrolysis.

Experimental design

One hundred and thirty 14d-old unsexed broilers (45 birds/treatment) were used in a 3 x 3factorial design. The first factor was the nucleotide supplement added to the feed in three levels: 0, 0.5g/kg, and 1g/kg feed up to 35 days of age. The second factor was the environment: hot environment with ambient temperature 32±1°C, comfortable environment with ambient temperature 23±1°C, and natural environment with an ambient temperature depend on the natural environment which ranged from 22 to 25°C in the morning and evening, and 30 to 35°C in the daytime.

A required quantity of each feed ingredient was purchased from the local market and the diets were prepared on the farm.

The nucleotides were added to the basal feed at graded levels, according to the experimental levels, and mixed by using homogenizer mixer.

Throughout the entire duration of the experiment feed and water were provided ad-libitum; lighting was continuous. The chickens were vaccinated against common viral diseases; Newcastle disease and Gumboro.

Data collection

- Body weight gain

The body weight of the experimental birds was recorded initially and on weekly basis.

- Feed intake

The feed was offered once daily, and the refusal feed collected and weighted at the next day.

- Feed conversion ratio (FCR)

The feed conversion ratio for each treatment-group was calculated by dividing the mean weekly total feed intake by the mean weekly total body weight gain (FI/WG).

Immune response

- Haemagglutinationinhibition (HI):

The antibody level against Newcastle disease as humoral immune response was determined by haemagglutinat ioninhibition (HI) test (OIE 2008), Two ml of blood was collected after 7^th days post ND vaccination. Forty-five blood samples were collected from 45 birds, 15 from each treatment. Blood samples were taken from the jugular vein in heparinized tubes and immediately centrifuged at 115 g for 15 min to obtain the serum. Then anti-body titrations were evaluated using a microtiter hemagglutination inhibition (HI) test.

- Immune organs

Relative weights of the spleen and bursa of fabricius were obtained as immunological indices at the end of the experiment (35 days of age).

Meat quality

The breast muscles (without skin) were used for physical quality analysis; pH, color, cooking loss, water holding capacity, and iron content as follows:

- pH

The pH was determined using a digital pH meter directly in breast muscle. At 24h post- mortem, pHu (ultimate pH) was determined by inserting the electrode directly into the muscle, as moisture spreads into the extra-cellular spaces (Mead 2004).

- Color

The color values of L*(lightness) a* (redness) and b* (yellowness) were determined using a spectrocolorimeter. For each sample, four readings were taken in breast muscle and the mean calculated for each sample (Mead 2004).

- Cooking loss

Cooking loss was determined in breast samples in an oven pre-warmed to 170°C. Crude breast muscle samples were weighed and put in trays with aluminum grills previously dried in an incubator. The trays were placed inside the oven until the sample core temperature reached 75°C. Samples were cooled at room temperature, re-weighed and cooking loss calculated as the difference between the initial and the final sample weights (ERL et al 2003).

- Water holding capacity

This assessment was based on the measure of water loss when pressure applied to the muscles. Meat cubes of 0.5g were placed between two filter papers and two glass plates, and a 35 kg-weight placed on the top glass plate for 5 minutes. The difference in breast muscle weight before and after the procedure represents the water loss. This was generally expressed as a percentage of the initial weight (ERL et al 2003).

- Iron analysis

The iron levels in breast muscle samples were conducted using a method described by Ma at el (2012). The samples (5 g) were dissolved in9ml concentrated HCl (37%) and 3 ml HNO₃ 65%, then heated at 100°C for 30 minutes. After, it was filtered, the filtrate taken, and then distilled water added until the volume became 50ml. The atomic absorption spectroscopic determination of iron was carried out with a spectrophotometer.

Statistical analysis

All the observations were analyzed by the General Linear Model (GLM) procedure of SPSS version 19.

The model used for data analysis was:

y_ijk= μ + A_i + B_j+(AB)_ij+ ε_ijk…………………………..(1)

Where:
y_ijk= observation k in level i of nucleotide and level j of environment,
μ = the overall mean,
A_i = the effect of level i of nucleotide,
B_j= the effect of level j of environment,
(AB)_ij= the effect of the interaction of level i of nucleotide with level j of environment, ε_ijk= random error.

Results and discussion

Effects of nucleotide supplementation and environmental conditions on performance parameters of broiler chicks are shown in Table 2. Overall, no an interaction between nucleotide supplementation and environment were observed for the feed intake, boy weight gain and FRC. However, based on the main effect, the addition of nucleotides to broiler feed did not affect feed intake, while continuous high temperatures resulted in reduced feed consumption. Highest feed intake was recorded in the natural environment where temperature was high during the day and low during the night and morning. This may be due to an increase in feed intake during low temperatures to compensate for daytime deprivation (Daghir et al 2009).

Broilers fed diet supplemented with nucleotide of 1g/kg had higher weight gain compared with 0.5g/kg and control group (Table 2). The hot environment reduced the growth rate. Supplementation of nucleotides improved the FCR. Supplementation of nucleotide at 1g/kg feed improved body weight gain and FCR in chickens reared under the hot environment.

Our study showed that the nucleotide supplementation improved the body weight gain and feed conversation ratio throughout the experimental period. This is in agreement with Esteve-Garcia et al (2007) who reported that addition of nucleotide at 500 mg/kg feed significantly improved body weight and feed to gain ratio at 21 days. Daneshmand et al (2017) also reported that the combination of commercially available nucleosides (adenosine, guanosine, uridine and cytidine) at level of 1g/kg significantly increased growth rate and improved FCR. Owens and McCracken (2007) also demonstrated that yeast extract supplementation to a broiler diet had a beneficial effect on feed intake and BW gain from 7 to 14 d of age. These results differ with those of Pelícia et al (2010) who found that the addition of nucleotides to broiler feed by 0.05, 0.06 and 0.07% did not have any effect on the broiler performance or carcass yield.

The positive results may be due to the increased availability of nucleotide for the proliferation of intestinal cells, which result in better digestion, and absorption of nutrients; this hypothesis is reinforced by many studies. Jung and Batal (2012) reported that supplementation of broiler feed with nucleotide (0.25% Torula yeast RNA and 2% Nupro® improved villus height and villus height-to-crypt depth ratio. Jackson et al (2018) found that Supplementation of rat diet with RNA (as source of nucleotide) significantly increased the mitotic index and lowered the PCI/MI ratio, indicating the hepatocytes were progressing through S phase into mitosis at a more normal rate. The results presented by Sato et al (1999) suggest that a nucleotide supplement may enhance enterocyte proliferation and/or maturation in vivo and in vitro.

Based on the interaction between nucleotide × environment, the inclusion of nucleotide in feed improved HI titer against ND in the different environmental conditions compare with control group (Table 3). Our finding is in agreement with Haldar et al (2011) who reported that the diet supplementation with yeast protein concentrate (YPC) and YPC-pellets improved immune response against Newcastle disease, may be due to the role of Nucleotides in modulating the cell mediated immunity and the T-cell dependent antibody responses (Jyonouchi 1994). The relative weights of bursa were affected (P < 0.05) by nucleotide × environment interaction, so that nucleotide supplementation was more effective in heat-stressed chicks, especially at the level of 0.5 g/kg of diet. Based on the main effect, supplementation of nucleotide increased the weight of the bursa of ferocious compared with control group but had no effect in spleen weight (Table 3). Environmental conditions had no effect on immune organ weight. This result is consistent with previous researches(Daneshmandet al 2017) indicating that adenosine increased the relative weight of the bursa of fabricius and had no effect on weight of the spleen. This may be explained that during stress, the rate of cell turnover in organs such as bursa increase, which demands sufficient amounts of nucleotide to synthesize DNA and RNA for maintenance and growth, and that exogenous nucleotide supplementation led to increase in the relative weight of the bursa. Previous studies have confirmed the absorption of exogenous nucleotides in the intestinal lumen and then migrate to immune organs such as the bursa (Hess and Greenberg 2012).

Based on main effects, the addition of nucleotide to the diet increased the Fe content in breast meat compared with control treatment, but had no impact on the other carcass quality aspects of broilers in this study (Tables 4 and 5).There were no interactions observed between nucleotide and environment on the cooking loss, PH and Fe content, while the WHC improved under hot environment by inclusion nucleotide to feed. This result in agreement with Chiofalo et al (2011)who reported that the nucleotide had no affect on the pH, lightness, and cooking loss, but increased iron content and redness value. The increase of iron content in meat can be explained by the role of nucleotides in the absorption of iron in the intestine through the conversion of nucleotides (AMP and GMP) to inosine, hypoxanthine, and uric acid, which increase the absorption of iron (Cosgrove 1998). Faelli and Esposito (1970) demonstrated that Inosine increased the intestinal absorption of iron in rats.

The hot and natural environment decreased the meat pH compared with other environmental conditions; the pH decline may be due to the conversion of glycogen to lactic acid in muscle(Zaboli et al 2019). The different environmental conditions had no significant impact on WHC, cooking loss, and color of broiler meat in this study (Tables 4 and 5). However, the interaction between nucleotide and environment had shown that the yellowness significantly increased under hot and natural environment by inclusion of nucleotide on level 0.5 g/kg.

Conclusion

This study concludes that nucleotides improved the weight gain and decrease FCR, but did not affect the feed intake. The best performance under heat stress was in birds fed nucleotides 1g/kg. The nucleotides improved the immune response of chicken under hot environment. There was no effect of nucleotides on pH, WHC, cooking loss and color, but increased iron content in meat. The level of 1g/kg appeared to be the best level of the nucleotide to reduce the harmful effects of high temperature in broiler performance and immune response.

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Received 27 July 2019; Accepted 26 September 2019; Published 2 November 2019

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