Performance of broiler Japanese quail supplemented with single-strain and multi-strain bacteria

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1 B-828 [1-6] Indian J. Anim. Res., Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Performance of broiler Japanese quail supplemented with single-strain and multi-strain bacteria M. Azhar 1, S. Ahmed 1 *, S. Mehmood 2, S. Naveed 1, S. Ahmad 2, M. Usman 2 and M.W. Zia 3 Department of Animal Nutrition, University of Veterinary and Animal Sciences, Lahore-54000, Pakistan. Received: Accepted: DOI: /ijar.B-828 ABSTRACT The growth performance of six-hundred (600) growing Japanese quails,distributed according to completely randomized design into 3 treatment groups(5 replicates/ treatment of 40 birds each) was recorded to evaluate the effect of feeding single and multi-strain bacteria. The experiment lasted for 4 weeks. Significant(P 0.05) results were observed regarding weight gain, folds of weight gain and FCR in the group fed multi-strain bacteria. Improved(P 0.05)total tract protein digestibility was detected in birds fed the diet containing multi-strain bacteria. Higher blood glucose level (P 0.05)was noticed in the group fed with single-strain bacteria.blood cholesterol level inbirds fedsingle and multi-strain bacteria was found higher (P 0.05). Enhanced(P 0.05)carcass characteristics and dressing percentage were recorded in birds fed with single strain bacteria.superior (P 0.05) weights of bursa and spleen wererecordedin the group fed with single-strain bacteria. Key words: Carcass yield, Digestibility, Growth, Multi-strain bacteria, Serum chemistry, Single-strain. INTRODUCTION The quail farming is attractive due to its highly productive and reproductive potential with a short incubation period of just 17 days. The quails are the smallest species of game bird which is farmed as well. These are found in the wild environment of Europe, Asia, America, and Australia but their commercial strains are raised for meat and eggs worldwide. The most common species of quails are Japanese quail (Coturnix japonica) which is used in commercial enterprises. Japanese quails are among the best meat producing quail in the world which is able to gain an average live weight of about 200 grams at four weeks of age. The reported weight of this particular species outside of production unit is claimed about 100 to 160 grams (Ahmad, 2014). In the modern era, the concept and application of probiotics are no more mystifying.the addition of antibiotics growth promoters though could enhance the growth performance of the animals but with increasing alarms regarding antibiotic resistance, the ban on subtherapeutic antibiotic usage, interest has been increased in finding alternatives to antibiotics and using probiotics is an approach that has potential to decrease enteric disease in poultry and subsequent contamination of poultry products (Elijah and Ruth, 2012). The probiotics are routinely used in poultry(lutful Kabir, 2009; Hanamanta et al., 2011; Walkunde et al., 2011) as well as fish (Archana and Pandey, 2013) diet as growth promoters. In this scenario, prebiotics, probiotics and symbiotic are generally used in poultry diet and considered beneficial for avian gut (Patterson and Burkholder, 2003). Probiotics are microbial cell cultures of live protective microorganisms that significantly affect the host by competing forother microorganisms (Shashidhara and Devgowda, 2003). These microorganisms improve gut health, increase appetite and improve the process of digestion and nutrient absorption. By using probiotics that attach to the mucosa ofthe intestine and make a barrier for harmful bacteria and enhance immune system (Dankowiakowska et al., 2013). Probiotics could be successfully added to poultry diet for growth, production, reproduction, modulation of intestinal microflora, immunomodulation, pathogen inhibition and promoting meat quality (Lutful Kabir, 2009).The global human population is increasing day by day and the people of developing countries are suffering from the hazards of malnutrition connected to protein foods especially from animal origin.the stated condition, therefore, demands concentrated efforts to produce theanimal protein in considerable quantity to fulfill requirements of the masses.current experiment was planned with the objectives to investigate the impact of the addition of single-strain and multi-strain bacteria in the diet of growing Japanese quails on growth parameters. MATERIALS AND METHODS A total of 600 d-old birds were procured from Avian Research and Training (ART) Center Hatchery, the University of Veterinary and Animal Sciences (UVAS), *Corresponding author s saeed.ahmed@uvas.edu.pk 1 Department of Animal Nutrition, University of Veterinary and Animal Sciences, Lahore-54000, Pakistan. 2 Department of Poultry Production, University of Veterinary and Animal Sciences, Lahore-54000, Pakistan. 3 Department of Livestock & Dairy Development, Punjab, Lahore-54000, Pakistan

2 2 INDIAN JOURNAL OF ANIMAL RESEARCH Lahore, Pakistan. The experiment was conducted for the period of 4 weeks at ART Center, UVAS, Lahore. The experimental protocol was approved by the Animal Ethics Committee, UVAS, Lahore. Birds were equally distributed into three treatment groups having five replicates of 40 birds each according to completely randomized design treatments were consisted of single-strain bacteria (Bacillus subtilis, PB6 naturally occurring spore-forming microorganism, provided as a premixture having spore counts of CFU/ g)@ 100 gram/1000 kg and multi-strain bacteria (three strains of Bacillus subtilis 60 gram/1000 kg and a control group. The growth performance was recorded to evaluate the effect of single and multi-strain bacteria. At the end of the experiment, nutrient digestibility was measured and three birds from each replicate (3 15= 45 birds) were picked and slaughtered for their carcass characteristics and serum chemistry. Birds were reared at octagonal ( cubic ft.) housing, equipped with multi-deck cages designed especially for the quails. This housing was well ventilated. Broiler quail ration (starter crumbs) were provided throughout the experiment to all the birds according to the recommendation of NRC (1994) having CP 22% and ME 3000 Kcal/Kg, with the addition to it, probiotics were supplemented (Table 1). Freshwater was provided round the clock by using nipple drinking system. Feed intake was calculated on daily basis by subtracting feed refused from feed offered. Weekly body weight of the birds was recorded with the help of electrical weighing balance. Feed conversion ratio was calculated on weekly basis by dividing weight gain from feed consumed.at the end of the experiment, 45 birds in total{(3 birds from each replicate (3 15= 45)} birds were picked up randomly from each treatment and 3 ml blood was collected in marked test tubes fromthe jugular vein of each bird during slaughtering. With the help of centrifugation process, serum samples were extracted and stored at -20C for measuring blood glucose and blood cholesterol levels. Dressed weight of each bird was recorded with the help of electrical weighing balance. Dressing percentage was calculated by dividing dressed weight by live weight. Thigh meat yield percent was calculated by dividing thigh meat yield with live weight. Breast meat yield percent was calculated by dividing breast meat yield divided by live weight. Liver, gizzard, and heart weights were also calculated. Table 1: Feed formulation and nutrient composition of experimental ration Ingredient (%) Ingredient (%) Maize grain Dry Matter Rice polishing Crude Protein Soya bean meal Crude Fibre Canola meal NDF NaCl/Salt ADF Total M.E 3000 (Kcal) NRC (1994) Statistical data analysis: Collected data were analyzed through one-way ANOVA technique (Steel et al., 1997) using PROC GLM in SAS software. Treatment means were compared through Fisher s LSD test assuming following mathematical model: Yij= µ + Ti + ij Whereas, Yij= Observation of dependent variable recorded on i th treatment µ = Population mean Ti= Effect of i th treatment (i = 1, 2, 3; single-strain bacteria, multi-strain bacteria, control group) ij = Residual effect of j th observation on i th treatment NID ~ 0, 2 RESULTS AND DISCUSSION Growth performance:values given in Table 2 show that the highest (P 0.05) feed intake was observed in birds having control diet as compared to the birds fed single and multistrain bacteria, respectively. Contradictory findings reported increased feed intake in chicken fed with multi-strain probiotics compared with thecontrol group fed basal diet (Zhang and Kim, 2014). Other study indicated that the feed intake values of groups were similar and lacked significance with layer flock that fed with Saccharomyces cerevisiae (Saadia and Nagla, 2010). In another study it was reported that the supplementation of probiotics did not improve the chicken s feed intake (Mansoub, 2010) whereas Timmerman et al. (2006) reported inconsistent findings. It may be due to the type of diet ingredients. It was observed that significant differences in feed consumption and efficiency (Willis et al., 2007). The highest body weight gain was observed in birds having multi-strain diet followed by the birds having control diet and diet with single strain bacteria, respectively. Similarly, other study reported that an overall increase(p 0.05) in body weight gain was found in chicken fed with multi-strain probiotics compared with that in control group fed basal diet (Zhang and Kim, 2014). Another study Table 2: Effect of single and multi-strain bacteria on TTP digestibility and growth performance of Japanese quail (Means ± SEM) Treatment Feed intake(g) TTP digestibility (%) Weight gain(g) Folds of gain FCR Liveability(%) (Straight Run) Control ±34.05 a 55.30±2.96 b ±1.44 b 25.77±0.22 b 3.17±0.18 c 79.92±3.90 a Single strain bacteria ±07.77 b 39.65±2.62 c ±1.70 c 24.99±0.22 c 3.05±0.06 b 64.61±6.85 c Multi strain bacteria ±07.68 c 62.58±4.80 a ±0.63 a 26.16±0.00 a 2.38±0.04 a 76.50±1.43 b Note: Superscripts on different means within column show significant difference (P 0.05); TTP: total tract protein

3 also showed that probiotics did not have any significant positive effect on broilers body weight gain (Babazadeh et al., 2011). In another study, it was demonstrated that improve daily weight gain was shown by the supplementation of probiotics in birds (Mountzourious et al., 2010). However, it was shown in several studies that there is no significant effect of probiotics on Body weight gain (Edens, 2003), the B. subtilis B2A used in this study also does not seem to improve body weight gain. Better feed conversion ratio(p 0.05) was observed in birds having control diet and single-strain bacteria in their diet as compared to the birds having multi-strain bacteria in their diet. It was reported that probiotic (Saccharomyces cervisiae) supplementation of broilers, atthe level of 1, 1.5 and 2%, had significantly improved the feed conversion ratio (Shareef and Al- Dabbagh, 2009). However, contradictory findings also reported that the dietary inclusion of probiotic did not affect feed conversion ratio at 42 days of age (Khosravi et al., 2008). In another study,it was also reported that the chicken broilers fed with protex in,the most advanced multi-strain probiotic in the world today that consists of seven different naturally occurring bacteria, primarily of a single strain of bacteria. The effectiveness and synergy of these strains have been proven in years of trials and production application had the lowest feed conversion ratio(rozbeh, 2016). It was found that the feed conversion ratio could significantly (P 0.05) improved by the use of Bacillus based probiotics at a specific concentration in the diet (cfu/g) in Guangxi yellow chickens (Zhou et al., 2010). The highest value of total tract protein digestibility was observed in birds having multi-strain bacteria in their diet and control diet as compared to the birds fed single-strain bacteria in their diet. It was also found that the probiotic L. bulgaricus could improve apparent ileal digestibility of crude protein in broiler chicken fed a maize-soybean-based diet (Apata,2008). Similarly, it was concluded that the probiotic E. faecium increased blood calcium levels in Vencobb broiler chicks, indicating improved bioavailability (Chawla et al., 2013). Probiotics can improve nutrient digestibility in poultry, but the interaction with different feed stuffs used in poultry diets is poorly understood at present. It was analyzed that digestion, absorption, and availability of nutrition improved by supplementation of probiotics accompanying with positive effects on intestinal activity and increasing digestive enzymes (Edens, 2003). Nutrient digestibility and caecal microflora Vol. Issue, () composition are positively affected by the use of probiotics in broiler chicken (Apata, 2008; Mountzouris et al., 2010). Blood serum chemistry: Table 3 shows that the highest (P 0.05) serum glucose level was observed in birds fed single-strain bacteria in their diet as compared to birds fed control diet and diet containing multi-strain bacteria, respectively. Blood Glucose level was increased (P 0.05) showing linear positive effect to probiotic supplementation. These results could be demonstrated by a higher absorptive capacity of the intestinal mucosa and it is because of histomorphological changes (Awad et al., 2009) and it is more effective digestion of the diet due to higher intestinal enzyme activity (Jin et al., 2000). The highest value of blood cholesterol level was observed among male birds fed multi and single-strain bacteria in their diet. It was reported that the level of cholesterol in serum significantly decreased in groups supplemented with probiotics and in assimilation of cholesterol by Lactobacillus compared to control group fed with basal diet (Mansoub, 2010). It was also investigated that a mixture containing 12 strains of Lactobacillus as probiotic decrease cholesterol level as compared to control group broiler chickens. It was reported in another study similarly with the previous one that a significant decrease in the serum level of triglycerides was observed between control group and groups treated with Lactobacillus acidophilus and Lactobacillus casei supplemented in broiler diet in combination with water or alone. Carcass characteristics: The values presented in Tables 4 and 5 show that in this experiment non-significant differences (P 0.05) were observed regarding relative bursa and spleen weight of Japanese quail fed with single and multi-strain bacteria as well as in dressed weight among male, female and straight run birds fed single and multi-strain bacteria in their diet. It was observed that non-significant difference in terms of slaughter performance were noted in hens having probiotics in their diet (Vali, 2009). Similarly, it was also reported that carcass yield did not increase significantly with the dietary inclusion of probiotics (Anjum et al., 2005). However, it was also noted that carcass yield increased in broilers having diets containing probiotics (Kavyani et al., 2012). The maximum value of dressing percentage was observed in male birds fed single-strain bacteria as compared to the birds fed multi-strain bacteria and control diet, respectively. Regarding female birds, the maximum value of dressing percentage was observed in birds group fed Table 3: Blood glucose, blood cholesterol and serum biochemistry of Japanese quail fed with single and multi-strain bacteria (Means ± SEM) Treatment Blood glucose (mg/dl) Blood cholesterol (mg/dl) Male Female Straight Run Male Female Straight Run Control ±15.06 a ±13.80 b ±8.21 b ±4.91 b ±4.26 b ±7.54 c Single strain bacteria ±23.41 b ±06.43 a ±9.29 a ±7.39 a ±4.87 a ±5.02 a Multi strain bacteria ±05.05 b ±11.12 b ±2.73 a ±8.10 a ±9.07 a ±6.18 b Note: Different alphabets on mean within column show significant difference (P 0.05)

4 4 INDIAN JOURNAL OF ANIMAL RESEARCH Table 4: Relative Bursa and spleen weight of Japanese quail fed with single and multi-strain bacteria (Means ± SEM) Treatment Relative Bursa weight (%) Relative spleen weight (%) Male Female Straight Run Male Female Straight Run Control 0.06±0.01 a 0.08±0.01 b 0.07±0.01 b 0.11±0.05 a 0.05±0.00 b 0.08±0.02 a Single strain bacteria 0.07±0.01 a 0.14±0.02 a 0.11±0.01 a 0.05±0.01 b 0.10±0.02 a 0.08±0.01 a Multi strain bacteria 0.09±0.01 a 0.08±0.01 b 0.09±0.01 ab 0.09±0.01 ab 0.07±0.01 ab 0.08±0.01 a Note: Different alphabets on mean within column show significant difference (P 0.05) Table 5: Effect of single and multiple strain on carcass characteristics of broiler Japanese quail (Means ± SEM) Parameters Control Single strain bacteria Multi strain bacteria Live weight ±3.43 b ±4.35 a ±3.83 c Dressed weight ±2.07 b ±3.07 a ±2.41 c Dressing (%) 57.55±0.25 b 58.61±0.63 a 57.56±0.40 b Thigh Meat Yield (%) 19.34±0.34 b 20.22±0.56 a 18.82±0.27 c Breast Meat Yield (%) 34.91±0.75 b 35.19±1.09 a 33.11±0.89 c Liver Weight (%) 2.55±0.19 b 2.70±0.16 a 2.54±0.17 b Gizzard weight (%) 3.10±0.15 b 3.30±0.21 a 2.80±0.11 c Heart weight (%) 0.88±0.02 a 0.84±0.02 b 0.82±0.02 c Intestinal weight (%) 4.67±0.21 b 5.52±0.37 a 4.16±0.17 c Intestinal length (cm) 64.25±0.89 c 72.22±0.74 a 70.06±1.53 b Note: Alphabets on different means within row show significant difference (P 0.05) control diet and diet containing multi-strain and single strain bacteria, respectively. However, non-significant difference was also observed in straight run birds. Non-significant effect of probiotics on dressing percentage in broilers were reported (Midilli et al., 2008). In similar fashion nonsignificant results of probiotics and control diets were observed in male birds in terms of dressing percentage (Bozkurt et al., 2009). In contrast to Banday and Risam (2001) and Hossain et al. (2008) reported positive effects on dressing percentage and carcass meat in the birds having probiotics and mixed antibiotics in their diet. In the recent study non-significant differences (P 0.05) regarding thigh meat yield percentage were observed among male, female and straight run birds fed, single and multi-strain bacteria in their diet and control diet. Regarding thigh meat yield, nonsignificant effect of probiotics and control diets on thigh yield percentage were found in broiler birds (Chumpawadee et al., 2008). In another experiment, Islam et al. (2004) observed non-significant effect of probiotics on thigh meat yield percentage. In current experiment non-significant differences (P 0.05) in terms of breast meat yield percentage were observed among male, female and straight run birds fed, single and multi-strain bacteria in their diet and control diet. Non-significant results regarding breast meat yieldpercentage might be due to non- significant effects in dressingpercentage. Non-significant results of probiotics and control diets on breast meat yield percentage in birds were reported Chumpawadee et al. (2008). However, In contrast to these studies Kabir et al. (2004) observed higher carcass meat with improved breast yield in broilers having diet containing probiotics and antibiotics through water and feed, respectively. In this study non-significant differences (P 0.05) regarding Liver weight percentage were observed among male, female and straight run birds fed, diet containing single and multi-strain bacteria and control group, respectively. In contrast to the findings of Ghahri et al. (2013), Liver weight was not affected by the use probiotics in diet. Ghahri et al. (2013) demonstrated that the supplementation of protexin in broilers diet had significant effects on liver weight. Azadegan Mehr et al. (2007) have indicated significant decrease in liver relative weight in male broilers having diet containing probiotics supplementation. Table 6: Economics of quail production reared under different treatments Parameters Single-strain Multi-strain Control group Feed consumed (g) Cost of day old chick (PKR) Total feed cost (PKR) Miscellaneous cost (PKR) Cost of feed additives/bird (PKR) Total cost/quail (PKR) Total live weight/quail (g) Sale price/ Kg live weight (PKR) Total sale price/quail (PKR) Net Profit/quail (PKR) Profit (%) PKR: Pakistani Rupee

5 In this experiment non-significant differences (P 0.05) in terms of Gizzard weight percentage were observed among male, female and straight run birds fed, single and multistrain bacteria in their diet and control diet, respectively. In different growth promoters, significantly higher Gizzard percentage were found in male Japanese quails fed on neomycin and as compared to protexin and control diets. In contradictory to the present study, Toghyani et al. (2011) and Islam et al. (2004) showed that the birds having diet with probiotics and antibiotics supplementation did not show any variation in the Gizzard percentage. In current study non-significant differences (P 0.05) regarding Heart weight percentage were observed among male, female and straight run birds fed, diet having single and multi-strain bacteria and control group, respectively. Heart weight percentage was not affected by the dietary probiotics (P 0.05) this finding is similar with that of Azadegan et al. (2014). Economics: The economics of quail production reared under different treatments is detailed in Table 6 which shows that Vol. Issue, () better (P 0.05) net profit/ quail (PKR 12.97) was noticed in the birds of that group which was fed with the diet supplemented with multi-strain bacteria than those reared on single-strain (PKR 3.07) added diet or control diet (PKR 3.12). Also, on overall basis the birds taken multi-strain bacteria in their diet presented the highest percentage (P 0.05) of profit compared to those birds received rest of the diets. CONCLUSION It can therefore be concluded that the diet supplementation with multi-strain bacteria had a positive influence on the growth performance, carcass traits, blood serum chemistry as well ason the economics of broiler Japanese quails. 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