Introduction

Formulating broiler diets on a digestible amino acid basis and utilizing the economically feasible commercial amino acid supplements (i.e. methionine, lysine, and threonine) results in diets marginally reduced in crude protein that support equal broiler growth to diets containing higher crude protein and excess amino acids (Kidd et al., 2002). The rapid growth rate of the present-day broiler requires increased amounts of all nutrients and energy on a daily basis, but these demands for different nutrients are not in the same proportions as previous ones. Amino acid (AA) requirements increase proportionately faster than do energy requirements; thus, a higher AA-to energy ratio is required in faster growing strains of broilers (Gous, 2010). Dietary AA concentrations should match needs for both maintenance and skeletal muscle accretion to effectively allow for increased synthesis of white meat (Kid et al., 2004). Benefits of diets with an increased AA density have been shown when applied early in life as well as in the finisher phases. Feeding diets containing high AA concentrations can result in greater economic re-turns if implemented in the starter phase, a period when feed intake is relatively low and growth rate is high (Kid et al., 2005), increasing Lys and other essential AA only in the finisher diet has also been shown to improve FCR and BMY (Dozier et al., 2007), compared with diets with average AA densities.

Administration of certain vitamins, minerals, amino acids and their different combinations to chicken in excess of their supposed requirements enhances their disease resistance. Vitamin E and selenium are one of them. Vitamin E plays important role in enzyme system in the animal body. Vitamin E added to levels beyond those needed to support optimal growth is beneficial in improving the immune-competence of growing broilers (Erf et al., 1998). Rajmane and Ranade (1994) found that the inclusion of vitamin E and C together at 150 mg/kg and 200 mg/kg diet respectively helped in improving both the growth of chicken and their immune response to vaccination. Choct et al. (2004) reported that increased dietary selenium content markedly reduced feed conversion ratio as a result of significantly lower feed intake of birds and improved eviscerated weight. The selenium yeast improved body weight and feed conversion ratio in broilers(Mahmoud and Edens, 2005). The ban against using antibiotics as feed additives in poultry diets of many countries has led to an increase in research regarding alternative feed additives, including prebiotics such as mannan oligosaccharide (MOS) and probiotics like Saccharomyces cerevisiae. MOS is derived from mannans on yeast cell surfaces. The benefits of MOS are based on specific properties, including modification of the intestinal micro-flora, reduction in turnover rate of the intestinal mucosa, and modulation of the immune system in the intestinal lumen. These properties have the potential to enhance growth rate, feed efficiency, and livability in poultry species (Parks et al., 2001). Probiotics, such as Saccharomyces cerevisiae, are defined as non-digestible ingredients, and they have several modes of action: beneficial changes in gut flora with reductions in the population of pathogenic bacteria, lactate production with subsequent changes in intestinal pH, production of antibiotic-type substances, production of enzymes, competition for adhesion receptors in the intestine, competition for nutrients, reduction of toxin release, and immuno-stimulation (Sohn et al., 2000; Han et al., 2007; Yin et al., 2008). The present investigation was carried out to elucidate effect of combination of amino acid, Vit. E, Selenium, MOS and biotin nutritional formula supplemented through feed at different dose levels on performance of broilers.

Materials and Methods

Experimental Site

The experiment was conducted at the broiler unit of Department of Poultry Science, College of Veterinary and Animal Sciences, Parbhani, Maharashtra Animal and Fishery Sciences, University, Nagpur (Maharashtra), India.

Experimental Birds and Data Collection

The experiment was carried out on three hundred day-old `Vencob’ commercial straight run broiler chicks for a period of six weeks. The experimental straight run chicks were randomly distributed into three Treatments with one control group. Each treatment group was further divided into three replicates having 25 chicks in each replicate. The nutritional formula product was supplied by M/s. Praj Industries Limited, Praj House, Bavdhan, Pune – 411 021 (Nutritional formula (NF) product containing iso-leucine, luceine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, histidine, mannan oligosacchirides, vitamin E, sodium selenite and biotine). The treatment group A was control without nutritional formula. The treatment group B, C and D were supplemented with Nutritional formula @ dose rate of 250 g, 500 g and 750 g per metric ton of feed, respectively (Table 1).

Data were collected on weekly weight changes was determined by weighing the birds on weekly basis and weight gain was calculated by subtracting the weight of the previous week from that of the current week. The feed intake was determined by subtracting the left-over feed from the feed offered, while feed conversion ratio was calculated as average feed intake divided by average /weight gain. The mortality was recorded daily and the weights of all the dead birds were taken in order to minimize an error in feed conversion ratio.

Table 1: Experimental Design for housing of broilers with supplementation of nutritional formula at different dose levels

It was expressed as percentage mortality at the end of the experiment for corresponding treatment group. At the end of the experiment, three birds from each treatment group (one bird from each replicate) were randomly selected for carcass evaluation studies. Edible carcass yield was calculated by adding the weight of carcass, neck, giblet and expressed as percentage of edible carcass yield of live weight. The weight of abdominal fat was recorded by separating it from carcass and expressed as percent abdominal fat of the total carcass yield. For the meat quality parameters, the meat samples from three birds of treatment group were stored for TBA and drip loss.

Housing and Management

Before arrival of broiler chicks, the experimental pens, waterers, feeders, brooders and floor were cleaned, washed, disinfected and fumigated using formaldehyde and potassium permanganate. Immediately after arrival, chicks were provided with antistress through drinking water. The birds were reared on deep litter system of housing. All the groups were provided with similar environmental and managemental conditions throughout the experimental period. An identical and adequate feeding and watering space was provided to all the birds throughout the experimental period. The birds were given free access to fresh, clean and wholesome drinking water throughout the experimental period. The immunization against Ranikhet Disease (B1 strain) and Infectious Bursal Disease (IBD Intermediate strain) was carried out on 7^th and 14^thday, respectively, followed by booster doses of IBD and RD on 21^st day and 28^th day respectively, through drinking water.

Formulation of Experimental Ration

The good quality feed ingredients were procured from local market for preparation of experimental diets. The basal diet was prepared with maize and soyabean meal having uniform nutrient composition. Feeding was done in three phases as pre-starter (200 g/bird), starter (1000 g/bird) and finisher (2800 g/bird) for all treatment groups up to six week of age. The nutrient levels for preparation of experimental diets (Table 2) were used as per the specification of BIS (2007).

Table 2: Percent ingredients and nutrient of different dietary composition of pre-starter, starter and finisher rations

#The nutritional formula was supplemented in the diet as described in the treatment details *Trace mineral mixture :- Copper, ferrous, zinc, iron. **Vitamin Mixture:- Vit. A, D, E, K and Vit.Bcomplex (Ribiflavin, choline, pantothenic acid, niacin, pyridoxine,biotin, cynocobalamine).

Statistical Analysis

All the generated data were subjected to statistical analysis (ANOVA) by using complete randomized design by Snedecor and Cochran, (2002). The treatment means were compared by critical differences (CD) and analysis of variance.

Results and Discussion

Weight Gain

The mean cumulative weight gains at 6^th week of age for treatment A, B, C, D and E were 2347.17, 2386.97, 2425.35 and 2552.28g, respectively (Table 3).

Table 3: Effect of combination of amino acid, Vit. E, Selenium, MOS and biotin nutritional formula supplemented through feed on various parameters in broilers

Note: The means bearing similar superscript within row do not differ significantly; SE-Standard Error, CD-Critical difference, CV- Coefficient of variance, *P<0.05, **P<0.01, ^NS– Non-significant

The analysis of variance for weight gain revealed significant improvement in weight gain in diet supplemented nutritional formula. Highest body weight gain was observed for Treatment group D supplemented with 750g per ton of feed nutritional formula. The findings in present study clearly indicated that there was linear significant correlation with supplementation of Nutritional formula at different dose level and weight gain. Similar finding was reported by Gous (2010) who reported that higher amino acid to energy ratio was required in faster growing strains of broilers. Similarly, Kidd et al., 2004 revealed that dietary amino acid concentrations were needed for both maintenance and skeletal muscle accretion to effectively allow for increased synthesis of white meat. Vieira and Angel (2012) reported that increasing the Lys and essential AA in broiler diets in the last phases of production allowed compensation for BMY because of the continuous high allometric growth rate of breast muscle. Gains in performance were expected to result from increases in dietary AA density

Feed Consumption

The significantly(P<0.05) highest feed consumption was observed for treatment group D(Table 3) however, the differences were non-significant for rest of treatment groups. The significant difference was recorded for supplementation of nutritional formula in broiler diet for feed intake. It concluded that there was no any adverse effect of nutritional formula supplementation on palatability of feed. The findings in the present study are in accordance with Flemming et al.(2004). They reported that the birds fed with the diet containing MOS and the diet containing antibiotic showed higher feed intake during the different production stages. Bozkurt et al. (2008) observed that the feed intake was tended to increase by feeding MOS.

Feed Conversion Ratio

The significant (P<0.05) improvement in feed conversion ratio was observed for nutritional formula supplemented Treatment groups (B and D, Table 3). It indicated the enhanced feed utilization and better nutrient absorption due to the surfactant properties of content of nutritional formula in the body of broilers. These results are agreement with Dozier et al. (2007). They concluded that increasing Lysine and other essential AA only in the finisher diet had improved FCR compared with diets with average AA densities. Corzo et al. (2005) reported that the feed conversion was lower in all periods for birds fed the high density diets. Kamran et al. (2013) reported that the weight gain and FCR of birds fed mannan oligosaccharides diet were better (P<0.05) than control group. Aravind et al. (2001) also observed improvement in feed efficiency in broiler birds supplemented with 0.15 ppm of selenium and 75 ppm of vitamin E/kg in diet. Bobade et al. (2009) reported that supplementation of vitamin E and selenium in the diet significantly improved feed efficiency in broilers.

Mortality

The higher survivability was observed in nutritional formula supplemented groups as compared to control. Among nutritional formula supplemented group D recorded highest survivability (Table 3) as compared to all groups.

Edible Carcass Yield

The mean value for edible carcass yield with giblet at 42^nd day was highest for treatment group C followed by D, B & A. However, the differences were non-significant. Similar findings were observed by Corzo et al. (2005). They observed no effect for yield of carcass, wings, drums, or saddle when broilers fed high density diet. However, Choct et al. (2004) found that birds receiving organic Se in their diets had improved eviscerated weight, breast yield.

Abdominal Fat

The mean abdominal fat percent value for treatment group B was highest (2.188) followed by C, D and A, but the differences were non-significant (P<0.05) in treatment groups A and D. In contrast to present study, Corzo et al. (2005) and Dozier et al. (2007) reported that the high-amino acid (AA) density diets may lower abdominal fat deposition.

Thiobarbituric Acid (TBA) Value

The non-significant statistical differences were shown among the treatment groups for TBA values as per ANOVA. However, numerically lower TBA values were observed for nutritional formula supplemented groups indicating Nutritional feed formula supplementation did not having antioxidant property. However, Naik et al. (2015) reported that the supplementation of organic selenium and vitamin E reduces the lipid peroxidation and autolysis, and improved the shelf life of broiler meat. Similar to present study findings, Hooge (2007) observed that TBARS values were similar to the control group when birds received diets supplemented with 0.30mg kg-1 of selenium in broilers.

Drip Loss to Present

There was numerically lower drip loss value of meat for nutritional formula supplemented groups resulted in maintenance of integrity of cell wall indicated antioxidant property. It is also evidenced by lower TBA values of meat stored at different age. Similar findings were observed by Choct and Naylor (2004) reported that the water loss over a 24 hr period for the dressed birds tended (p=0.06) to be reduced as the dietary vitamin E level was increased from 50 IU to 100 IU. Increasing the vitamin E content of the feed was reduced average drip-loss from 1.19 to 1.02%.

Economic Return

Feeding diets containing high AA concentrations can result in greater economic returns if implemented in starter phase, a period when feed intake is relatively low and growth rate is high (Kidd et al., 2004), because some of the benefits appear to have a carryover effect throughout growth to market age (Halevy et al., 2000, Pophal et al., 2004). Corzo et al. (2005) reported that overall improvements in economically important parameters observed after feeding an amino acid concentration greater than standard nutritional programs.

Conclusion

Overall findings in the present study inferred that supplementation of nutritional formula plays significant role in improving the bird’s performance in term of body weight, feed consumption, FCR and meat quality parameters. It played important role in improving economics of broiler production. The nutritional formula supplemented at dose level of 750 gm/MT of feed is more economical due to fast growth and resulted in more profit.

Acknowledgement

The authors are thankful to the Associate Dean, College of Veterinary Science and Animal Sciences, Parbhani for providing necessary facilities to accomplish the work. The authors are also thankful to M/s. Praj Industries Limited, Praj House, Bavdhan, Pune – 411 021, for supply of nutritional formula product and sponsoring the project.

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