Introduction

The protein provides amino acids for tissue growth and egg production. The dietary protein requirement of layer is influenced by metabolizable energy content and the ingredients used to formulate the diets. Protein is the most expensive nutrient and must be provided from a high quality source with optimum quantity. Supplementation with synthetic amino acid in diets of poultry increases feed conversion efficiency, lowers feed cost per unit of weight gain or production, reduces nitrogen excretion and has other beneficial effects. Appropriate concentration of energy and amino acid in feed are needed in order to increase the productivity of laying hens Chan et al. (2003). Imbalance in amino acid composition in layer diet which is needed for egg protein synthesis will consequently result in lowered egg production along with poor plumage condition, cannibalism and increased mortality (Ambrosen and Peterson, 1997). The synthetic amino acid especially methionine is listed among the prohibited synthetic substances and its use has been questioned in organic farming practices because synthetic methionine is metabolized into highly toxic compounds and thereby, adversely affecting performance of poultry birds (Bender, 1975). Herbal methionine premixes, manufactured in India have recently found their way into animal feed industry. AV/CAP/18 is herbal amino acid premixes that are available in animal feed market. They are phyto-additive containing herbal ingredients that mimic the activity of methionine. Under Indian conditions, herbal methionine (Methiorep®) has been found to replace DL-methionine very effectively when used in broiler rations (Chattopadhyay et al., 2006; Kalbande et al., 2009). Hence, the present study was undertaken to determine the efficacy of herbal combined amino acid (AV/CAP/18 supplied by M/S Ayurvet Limited, Baddi, H.P, India) with synthetic amino acid on egg quality parameters. The herbal product consisting of herbs like Allium sativum, Mucuna pruriens and many others.

Materials and Methods

Nutrient Composition of Ingredients and the Treatment

Ingredients and Experimental Rations

The experiment was conducted at Department of Poultry Science, College of Veterinary and Animal Sciences, MAFSU, Parbhani (Maharashtra), India. Corn grains, DOC Soy, rice polish, groundnut cake, DORB, DCP, Vit-Mix., Min-mix ingredients were used for preparation of layer feed (Table 1). The treatment Group (T₀) was negative control with basal diet. This group was fed diet completely devoid of lysine and biotin and with partial deficiency of methionine and choline chloride, from 20^th -41^th week. Group T₁ was the positive treatment group fed basal diet supplemented with DL methionine@2kg/tone of feed, choline chloride@500, biotin@150mg/tonne and Lysine/@1kg/tonne offered. Group T₂ is the treatment group fed basal diet deficient in amino acids (T₀) and supplemented with AV/CAP/18@ 2kg/tonne of feed. Group T₃ was the treatment group fed basal diet deficient in amino acids (T₀) and supplemented with DL methionine@1kg/tonne of feed and AV/CAP/18@2kg/tonne of feed. Group T₄ is the treatment group fed basal diet deficient in amino acids (T₀) and supplemented with DL methionine@1kg/tonne of feed and AV/CAP/18@1.5kg/tonne of feed.

Table 1: Feed formulation, feed ingredients, mixing ratio for various treatment groups supplemented with herbal amino acid and synthetic amino acid

Management of Experimental Birds

A study was conducted on 240 BV³⁰⁰ commercial layer birds of 18^th week age; the birds were reared in California cages and were maintained under ideal conditions. Vaccination was done as per the recommended schedule. The birds were randomly divided into five groups of 48 birds with 3 replicates of 12 birds in each group (Table 2). The birds were having free access to feed and water.The birds were fed layer ration with 17% CP. Total feed intake per group during the experimental period of 20-41 weeks were recorded and feed efficiency was calculated.

Table 2: Experimental design for various treatment groups supplemented with herbal amino acid and synthetic amino acid

Data Collection and Measurement

Egg Quality Parameters

Egg quality parameters were measured for each replicate. The eggs gathered during time interval of 18^th – 40^th weeks were used to measure exterior and interior egg quality. The whole egg was weighed (g) on a sensitive weighing balance to determine average egg weight. Egg production, and egg mass was calculated based on average weight.The represented sample eggs were carefully broken out on a white tile (30 × 30 cm) with the blunt edge of a knife to separate egg contents and internal trades like the albumen, yolk, and shell thickness were measured. The albumen of the broken egg was carefully separated from the yolk using a separator and the albumen and yolk weights were recorded with a sensitive weighing balance. The egg shell was dried and weighed. From this data, albumin ratio and yolk ratio were calculated. Mortality was also recorded. Economics of each treatment was calculated on the basis of input cost.

Statistical Analysis

Mean and standard deviations were computed by applying FRBD. Economics of each treatment was calculated by simple mathematical calculations.

Results and Discussion

Egg Production Parameter

Weekly Egg Production

Treatment group T₂ fed basal diet deficient in amino acids (T₀) and supplemented with AV/CAP/18@ 2kg/tonne of feed revealed (Table3) significantly increased egg production.

Table 3: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplementation on egg Production

Note: – means in columns not sharing a common superscript differ significantly

The present reports found to be in agreement with Harms and Russell (1996) who also reported that supplementing methionine in the diet of laying hens increases the egg production. Similar findings were also reported by Manwar et al. (2016). They revealed that hen housed egg production in Methiorep supplemented group birds was significantly high (P<0.05) than the birds in control group birds. However, Igbasan et al. (2012) clearly revealed that layers fed on diets supplemented with HM (Meth-o-Tas®) produced less egg. The present study inferred that increase in egg production in low intake of amino acids with supplementation of herbal amino acids AV/CAP/18@ 200g/100kg of feed compensated egg production compared to treatment group (T1) indicating efficacy of herbal amino acid.

Egg Weight

The treatment group T₃ supplemented with DL methionine@1kg/tonne of feed and AV/CAP/18@ 2kg/tonne of feed significantly increased egg weight (Table 4).

Table 4: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on egg weight (20-22 week)

Note: – means in columns not sharing a common superscript differ significantly

There is evidence in the literature to demonstrate that fortification of laying hen diets with methionine increases egg weight (Shafer et al., 1998; Narvaez-Solatre et al., 2005; Liu et al.,2005). The increase in egg weight is supported by findings of Zeweil et al. (2011) who reported that egg weight increased with increasing level of amino acid intake by laying hens.

Egg Mass

Significantly (Table 5) increased egg mass was observed for treatment group (T₃) supplemented with DL methionine@1kg/tonne of feed and AV/CAP/18@ 2kg/tonne of feed. The egg mass was at par to that of positive control (T1). It indicated efficacy of the herbal amino acid compared to that of negative control group (T0), with lowest egg mass.

Table 5: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on egg mass

Note: – means in columns not sharing a common superscript differ significantly

Significantly highest egg mass were recorded from 31^_34^th week of age. The increased egg mass is supported by findings of Keshavarz (1992) and Jackson. Liu et al. (2005) who reported that egg mass was found to be having positive correlation with amino acid content in the layer diet. The increase egg mass was mainly contributed from increased egg weight.

Feed Cost per Egg

The treatment group (T2) supplemented with DL methionine @1kg/tonne of feed and AV/CAP/18@ 2kg/tonne of feed and treatment group T3 recorded significantly lower feed cost per egg(Table 6) compared to positive control group(T1). Similar finding was observed by Manwar et al. (2016). The results were found to be in agreement with Sharma Ranjan (2015), Pillai et al. (2006) who also reported that supplementation of Methionine increases the feed efficiency in poultry birds and thereby reduced cost.

Table 6: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on feed cost per egg (Rs/Egg)

Note: – means in columns not sharing a common superscript differ significantly

Feed Efficiency

The treatment group (T2₎ supplemented with DL methionine@1kg/tonne of feed and AV/CAP/18@ 2kg/tonne of feed and treatment group T3 recorded significantly better feed efficiency compared to positive control (Table 7). This may be due to the sophisticated relationship that has evolved between the GI tract and gut microbiota allows for efficient utilization of dietary nutrients. Similar finding was observed by Manwar et al. (2016). The results were found to be in agreement with Sharma Ranjan (2015), Pillai et al. (2006) who also reported that supplementation of Methionine increases the feed efficiency in poultry birds.

Table7: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on feed efficiency (20-22 week)

Note: – means in columns not sharing a common superscript differ significantly

Egg Quality Parameters

Albumin Weight

There were non-significant differences among the various treatment groups for albumin weight (Table 8). However, numerically higher values were observed for herbal amino acid supplementary group (T2) indicating important role of herbal amino acid, it compensated to that of positive control (T1). The highest values of albumin weight were recorded during 25^th and 32th week. The increase in albumen weight may be due to limited availability of amino acid having adverse effects on albumen solids (Shafer, 1998).

Table 8: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on albumin weight (gm)

Note: – means in columns not sharing a common superscript differ significantly

Yolk Weight

Herbal Combined Amino Acid Formula Av/Cap/18 and Synthetic Amino Acid Supplements on yolk weight revealed (Table 9) numerical increased yolk weight for herbal combined amino groups. The highest yolk weights were recorded in the treatment group T4.

Table 9: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on yolk weight (gm)

Note: – means in columns not sharing a common superscript differ significantly

It clearly indicated supremacy of supplementation of herbal alternatives of amino acid. The present findings are in accordance with Shafer, (1998) who reported that increase in yolk weight may be due to limited availability of amino acid having adverse effects on albumen solids.

Shell Thickness

Significantly higher shell thickness values were recorded in the treatment group T2 (Table 10) indicating numerical increased shell thickness for herbal combined amino acid group. Thus better shell strength and ultimately egg quality was obtained with herbal product supplementation. Similar, results were obtained by Hassan and Ragab (2009) in case of supplementation of Trigonella foenum graecum on productive performance and egg quality of layers.

Table 10: Comparative efficacy of herbal combined amino acid with synthetic amino acid supplements on shell thickness (mm)

Note: – means in columns not sharing a common superscript differ significantly

Mortality (%)

The highest mortality was observed for T0 (4.21) followed by T4 (2.5), T1 (1.95), T3 (1.88) and T2 (1.78). The lowest mortality percent was observed in herbal combined amino acid group (Formula Av/Cap/18). This clearly indicate that an adequate dietary all amino acids are necessary for normal immune-competence and protecting the birds from variety of the diseases as reported by Peng Li et al. (2007).The implication is that dietary herbal methionine has no detrimental effect on survivability of chickens. Rajurker et al. (2009) also reported that herbal methionine supplement (Methiorep®) is totally safe and has no adverse effect even when used at the highest limit dose of 5 g/kg body weight of male wistar rats.

Conclusion

The data from trial indicates that basal diet supplemented with herbal combined amino acid formula Av/Cap/18 significantly increased egg production, egg weight, egg mass, yolk weight and shell thickness. Similarly superior feed efficiency and reduced cost of feed per egg was observed. Hence, it is concluded that diet with herbal combined amino acid formula Av/Cap/18 @ 200g/100kg of feed of feed from 20^th to 41^th week is most beneficial for improving egg production, egg weight, egg mass and shell thickness. AV/CAP/18 formula can be one of the best alternative for synthetic amino acids, could successfully replace the synthetic amino acids for optimum production performance of laying hens.

Acknowledgements

The authors are highly grateful to Associate Dean, College of Veterinary and Animal Sciences, MAFSU, Parbhani, for providing necessary facilities and for his help at various stages of the experiment. The authors are also highly grateful to M/S Ayurvet Limited Post Baddi, Himachal Pradesh, for providing Herbal amino acid formula AV/CAP/18 sample and sponsoring the research trial.

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