COMPARATIVE STUDY ON EFFECT OF TURMERIC POWDER, PROBIOTIC AND PREBIOTIC SUPPLEMENTATION ON BROILER PERFORMANCE AND IMMUNITY

Assiut University web-site: www.aun.edu.eg

COMPARATIVE STUDY ON EFFECT OF TURMERIC POWDER, PROBIOTIC AND PREBIOTIC SUPPLEMENTATION ON BROILER PERFORMANCE

 AND IMMUNITY

L.K. ABD EL-SAMIE

Assistant Consultant of Avian and Rabbit Diseases, the Educational Vet. Hospital Zagazig Univ.

Received: 30 September 2019;     Accepted: 31 October 2019

INTRODUCTION

Gibson and Roberfroid (1995) described Prebiotics as non-digestible polysaccharides and oligosaccharides which boost the growth of beneficial lactic acid bacteria in the colon responsible for suppressing of Salmonella sp. or Escherichia coli multiplication. It is revised and categorized by Stowell (2007) to established prebiotics (Inulin, fructooligosaccharides (FOS), galactooligosaccharides (GOS), lactulose and polydextose) and emerging prebiotics (isomaltooligosaccharides (IMO), xylooligosaccahrides (XOS), and lactitol). Sabater-Molina et al. (2009); Xu et al. (2009) and Femia et al. (2010) reported that established prebiotic such as FOS (chicory root) and XOS (wheat bran) have enormous usages. Moreover Yeo and Liong (2010); Vamanu and Vamanu (2010) and Mandal et al. (2009) mentioned that mannitol, maltodextrin, raffinose, lactulose and sorbitol are prebiotics of wide

Corresponding author: L.K. ABD EL-SAMIE

E-mail address: lamahsamie@gmail.com

Present address: Assistant Consultant of Avian and Rabbit Diseases, The Educational Vet. Hospital Zagazig Univ.

health applications. Avian gastrointestinal tract contains different and dynamic population of microorganisms of cooperative relationship with its host nutrition, metabolism and immunity (Sohail et al., 2012). Probiotics described by FAO/WHO, (2001) as live microorganisms which deliberate health advantage when administered sufficiently by the host. Also known as direct-fed microbials, which classified as live nonpathogenic microorganisms that are capable of maintaining a normal gut microbial population by Patterson and Burkholder (2003) and Ohimain and Ofongo (2012). It include strains of Lactobacillus, Bifidobacterium, and yeast, indigenous in the colon of chicken, which enhance the levels of health by producing bacteriocins that suppress the growth of pathogenic bacteria as reported by Alavi et al. (2012). Probiotics can help in keeping a healthy balance of microorganisms via several mode of actions comprising competitive exclusion, pathogen antagonism, altering metabolism by increasing digestive enzyme activity and stimulation of the immune system, noted by Dierck (1989) and Cox and Dalloul (2015). Moreover Kaiber et al. (2004) and Pender et al. (2016) mentioned that probiotic may provide a potential alternative to the prophylactic use of drugs in food animals due to their studied abilities to reduce severity of enteric diseases and enhance performance in poultry. Amalraj et al. (2017) described Curcumin as yellowish powder derived from herbal medicinal plant turmeric (Curcuma longa) of numerous pharmacological uses antioxidant, antiprotozoal, antivenom, antimicrobial, anti-inflammatory, antiproliferative, antiangiogenic, antitumor and antiaging. Al Sultan (2003) reported that usage of Curcuma longa, turmeric, as feed additive at level of 0.5% boosted overall performance of broiler chickens with recommendation of cost effectiveness study.

The research aimed to investigate the influences of Prebiotics, Probiotics and Curcumin supplementation to broilers diet on their body gain, feed intake, feed conversion rate, hematological parameters and immune response.

MATERIALS AND METHODS

Ethical approval: This study was carried out under the authorization of the Animal Welfare and Research Ethics Committee, Faculty of Veterinary Medicine, Zagazig University, Egypt.

Birds: 150 of day old chicks (Arbor acer) were sourced from local hatchery, reared on floor pens and fed on commercial grower ration contain 22% protein and yield 3150 Kcal/kg (Table1).

Probiotic: Bacillus licheniformis 8 x 10⁹ CFU (Gallipro Tect^®) added to the feed as 100gm per ton according to the producer recommendation along the research period starting with the first day.

Prebiotic: Inactivatedsaccharomyces cerevisiae var. ellipoideus 1x10¹⁰ CFU (Thepax^®) supplemented as 100gm per 100kg feed according to the producer recommendation for the whole period started from the first day.

Turmeric powder: Curcumin was purchased from local market and mixed in the feed 7.5gm/kg according to Shohe et al. (2019).

Vaccines: Avian Influenza (AI) type A H9N2 (Cevac Flu H9K) inactivated oil adjuvant vaccine deployed for immunization of broilers at day old subcutaneous at the back of the neck with 0.2ml. Infectious Bursal Disease (IBD) strain MB5 10² EID₅₀ lyophilized live vaccine used for immunization of broilers at 8 days old via drinking water according to the manufacturer guidelines.

Samples: Blood were collected from wing vein with vacutainers needle from all groups at 3 and 5weeks old for hematological analysis by using heparinized vacutainer and normal vacutainer for immunological assay.

Experimental birds and design: Chicks randomly were divided equally to 5 groups (A negative control, B positive vaccinated, C vaccinated with Curcumin supplement, D vaccinated with probiotic supplement and E vaccinated with prebiotic supplement) and subjected to weighing and blood sampling at 3and5 weeks of age. Groups A, B, C and E were vaccinated with against AI and IBD

Performance assessment: Body weight(BW) of chicks was recorded individually at recipient day as well as feed supplement for each group to calculate feed intake (FI) thereafter BW and FI were logged at 3 and 5 weeks of age. Weight gain (WG) was calculated by the difference between BW of 35, 21 and 0day old while feed conversion rate (FCR) was calculated by dividing the amount of FI for a period by the BG of the same period.

Immuno-assay: Commercial kits were used for determination of IBD antibody titer (IDEXX) and (Bio Check) for AI. According to the manufacturer instructions, harvested serum was diluted and procedure was followed. Sample to positive (S/P) ratio was calculated as the difference between sample and negative control values divided by the difference between positive control and negative control values. At 1:500 dilution (Log₁₀) titer = 1.09 (log₁₀ S/P) + 3.36 for IBD while = 1.1 * Log (SP) + 3.156 for AI.

Hematological analysis: Hematology indicators such as erythrocyte count (RBCs), packed cell volume (PCV), white blood cell (WBCs), WBC differentials and mean corpuscular hemoglobin (Hb) were investigated. Hb concentration was measured using Van Slyke Apparatus, and PCV – Hacksley Hematocrit Centrifuge (UK). WBC and its differentials were determined using the Neubaer count chamber following procedure described by Fudge (2000) and Cray and Zaias (2004).

Statistical assay: Data were compiled and analyzed using one-way analysis of variance (ANOVA) through the general linear models (GLM) procedure of the statistical Package for Social Science version 20.0 (SPSS for windows 20.0 Inc., Chicago, IL,USA). Duncan multiple range test used to separate means at P<0.05.

RESULTS

Growth performance parameters were summarized in Table 2. Addition of Curcumin, Probiotic and Prebiotic resulted in upsurge of BW than control and vaccinated groups. At 21days old, the recorded BW were 868.83±8.53^a, 843.67±12.03^ab, 832±11.7^b,817.37±12.36^bc and 796.27±8.56^c for Curcumin, Probiotic Prebiotic, vaccinated and control respectively. Statistically BW difference was insignificant between Curcumin and Probiotic, between Probiotic and Prebiotic, Prebiotic and vaccinated and between vaccinated and control groups, while significant differences were recorded between Curcumin, Prebiotic and control.  At 35 days old, there was no significant difference between Curcumin, Probiotic and Prebiotic in BW 1999.4±22.74^a, 1999.4±22.74^a and 1959.73±24.08^a, respectively, but significant different with vaccinated and control group 1820.67±24.22^b and 1789.03±17.01^b, respectively. Overall FI was significantly lower for Prebiotic group 3371.77±30.70^b comparing with Curcumin 3452.53±18.56^a and vaccinated 3455.47±17.47^a while overall WG was not significantly varied within Curcumin, Probiotic and Prebiotic groups 1948.83±22.86^a, 1950.83±22.67^a and 1914.03±23.80^a, respectively but significantly higher than control and vaccinated 1750.60±16.47^b and 1772.20±24.75^b respectively (Fig1). Similarly overall FCR was 78±0.020^b, 1.76±0.025^b and 1.77±0.026^b respectively, but significantly better than control and vaccinated group (Table2) (Fig.2).

Hematological indices revealed that Curcumin and Probiotic at 21 and 35 days old improved significantly PCV, RBCs count and WBCs count than other groups (Table3). At 3 weeks old PCV values were 34.09±0.24^a, 34.08±0.24^a, 32.69±0.26^b, 32.03±0.26^b and 30.45±0.26^c, RBCs count were 2.49±0.017^a, 2.50±0.017^a, 2.40±0.017^b, 2.32±0.016^c and 2.20±0.014^d 10⁶/µl, WBCs count were 31.95±0.28^a, 31.91±0.28^a, 31.45±0.29^ab, 31.05±0.24^b and 23.49±0.25^c 10³/µl for Curcumin, Probiotic, Prebiotic, vaccinated and control group, respectively. Prebiotic had no significant effect on Hb concentration, but increased significantly RBCs and WBCs count than vaccinated and control group. Moreover at 35 days old all additives had no influence on Hb 8.99±0.104^a, 8.96±0.103^a, 8.64±0.096^a, 8.42±0.095^a and 8.31±0.097^a, respectively, but had significant effect PCV, RBCs and WBCs count.

Differential leucocytes showed significant higher count of lymphocyte 22.45±0.23^a, 22.42±0.23^a and 20.74±0.22^a, 20.71±0.22^a at 3 and 5 weeks of age in Curcumin and Probiotic respectively,(Table4). ELISA IBD antibody titer at 21 day old exhibited higher titer in groups supplemented with Probiotic (Fig.3)where it was 2374.73±19.44^a compared with Curcumin 2317.63±18.80^b, Prebiotic 2297.63±17.88^b, vaccinated 2243.77±20.42^cand control which showed negative titer. While Curcumin and Probiotic not only induced higher AI titer 1606.30±10.36^a, 1592.83±15.75^a, respectively, at 21 day old (Fig.4) but also at 35 days old (Fig.5).

Fig. 1: Influence of Turmeric, Probiotic and Prebiotic on overall Weight Gain

Fig. 2: Effectiveness of feed additives on overall Feed Conversion Ratio

Fig. 3: Post vaccinal reaction against Infectious bursitis

Fig. 4: Post vaccinal reaction against Avian Influenza at 3weeks of age

Fig. 5: Post vaccinal reaction against Avian Influenza at 5weeks of age

Table 1: Composition of grower basal diet

Table 2: Growth performance parameters of broilers at different experimental intervals.

A: control.      B: vaccinated.     C: vaccinated with Curcumin.     D: vaccinated with Probiotic.    E: vaccinated with Prebiotic.

BW: body weight.   FI: feed intake.        WG: weight gain.    FCR: feed conversion rate.

* Means ±standard error within the same row carrying different superscript are significantly different at P value<0.05.

Table 3: Hematological analysis of broilers supplemented with feed additives.

A: control.     B: vaccinated.     C: vaccinated with Curcumin.     D: vaccinated with Probiotic.     E: vaccinated with Prebiotic.

PCV: packed cell volume.                       WBCs: white blood cells.                        RBCs: red blood cells.

* Means ±standard error within the same row carrying different superscript are significantly different at P value<0.05.

Table 4: Immuno-assay of broilers supplemented with feed additives.

A: control.     B: vaccinated.     C: vaccinated with Curcumin.     D: vaccinated with Probiotic.     E: vaccinated with Prebiotic.

AI: avain influenza antibody.  IBD: infectious bursal disease antibody.             Hetero: heterophil count.             Lympho: lymphocytic count.

* Means ±standard error within the same row carrying different superscript are significantly different at P value<0.05.

AI titer < 600 consider negative.                                             IBD titer < 200 consider negative.

DISCUSSION

Excessive usage of antibiotics in poultry industry, subsidized the importance of herbal plant, Prebiotics and Probiotics convention. Therefore this study was conducted to investigate its effect on broiler production. Although addition of Curcumin, Probiotic and Prebiotic to basal diet had slightly affected overall FI, performance parameters as BW, overall WG and FCR of broilers had significantly improved (p < 0.05) on matching with the control and vaccinated groups. However, Curcumin had highest BW, WG and FI at 21 days old, there was no significant favor to any of these additives at the end study except Prebiotic had the lowest overall FI. Clockwise, Abdel-Hafeez et al. (2017) pointed that chicks fed diets supplemented with Probiotic, Prebiotic and Synbiotic (with and without feed restriction) showed higher BW and FCR than chicks fed the control diets. Moreover PCV was increased in additive treatments with restricted feeding at the end of the experiment. Alimohamadi et al. (2014) resulted that at 28 days old, diets mixed with Black seed, Cumin seed and Probiotic increased body weight (p = 0.027) with no significant differences in final body weight and average daily feed intake among any dietary treatments (p > 0.05). Though the chicks fed diets mixed black seed, Probiotic and Prebiotic exhibited significantly better FCR than chicks fed the control diet during the entire experimental period (p = 0.048). Tayeri et al. (2018) noted that there was no effect of Synbiotic, Probiotic, Prebiotic and antibiotic treatment on FI. Yet, weight gain was greater for broilers (p = 0.001) fed Synbiotic (73.6 g/d), Probiotic (72.8 g/d) and prebiotic treatments (69.8 g/d), when compared with broilers fed the antibiotic (66.3 g/d) or no supplement (64.2 g/d). In the same line, Rajput et al. (2013) documented that, at 42 days old (finisher phase) live body weight (P＜0.0003) and FCR (P＜ 0.0172) were significantly enhanced in group that received 200mg/kg diet Curcumin, as compared to control and other groups. Also, these findings agreed with Kafi et al. (2017), Ahlawat et al. (2018) and Shohe et al. (2019) who recorded that not only the average BW and BG was significantly (P <0.05) the highest in group which supplemented with turmeric powder at the rate 7.5g/kg feed but also the value of FCR was the lowest. Improvement of growth and feed conversion efficacy may be explained by indorsement of a well-balanced gut microflora in early life which can be established by feeding of Probiotic/Prebiotic in the starter diet (Salim et al., 2013) and/or modulation of microbial population by phytogenetic products (Windisch et al., 2008). Also stimulation of gastric and pancreatic digestive enzymes can be accredited by spices and their derivatives (Srinivasan, 2005) that finally lead to more absorption of essential nutrients.

Hematology indices reflected highest influences (p < 0.05) of Curcumin and Probiotic supplementation on PCV, RBCs and WBCs values at 3 and 7 weeks old as well as Hb concentration at 3weeks old. This positive effect may attributed to anti-inflammatory, antioxidant properties of Curcumin and digestive enzyme intensifying of Probiotic that enhance metabolism consequently improve iron absorption and utilization leading to enhancement of RBCs production and Hb concentration consequently PCV. Similarly, Beski and Al-Sardary (2015) who noted that Probiotics and Synbiotics resulted in a significant increase in the concentration of Hb, as well as, Alimohamadi et al. (2014) who declared that RBC counts, hemoglobin concentration and hematocrit percentage were significantly higher in the chicks fed diets contain Black seed compared with those fed the control diet (p < 0.05). Furthermore, Salim et al. (2013) recorded that WBC count was significantly higher in chicken fed on diet contain 0.1% mixture of Probiotic and Prebiotic (DFM2) compared with the other contain 0.1% virginiamycin and control-fed birds, likewise monocyte level was higher in DFM 2. Contrarily, Kafi et al. (2017) and Shohe et al. (2019) found that no significant difference in Hb and PCV values of broiler birds when supplemented with turmeric powder irrespective of levels. Moreover Abdel-Hafez et al. (2017) noted that there were no statistical differences (p>0.05) in Hb and PCV% between the control and the other groups which received Synbiotic, Probiotic, Prebiotic and Adegoke et al. (2018) mentioned that broilers received basal diet with Curcumin at level of 400g/100kg in ad libitum feeding, had no significant alternation on PCV, RBCs and Hb except WBCs.

Immune response analysis exhibited boosting of Curcumin, Probiotic and Prebiotic to vaccines reaction for both AV and IBD. Likewise, Hong et al. (2005) reported that Bacillus-based direct fed microbals (DFM) enhances immune function and promotes the synthesis of endogenous antimicrobial peptides in the gut. In addition Janardhana et al. (2009) highlighted that Prebiotic fructooligosaccharide supplement increased IgG and IgM in broilers. Present findings provoked highest ELISA antibody titer at 3weeks old for AV was recorded in Curcumin and Probiotic group but for IBD in Probiotic. This variation could be justified by the type of vaccine, age of vaccination and the route of administration.

Results could be concluded that addition of Curcumin, Probiotic and Prebiotic to the broiler’s diet had performance enhancement and immune stimulant properties and improved overall body weight/gain and feed conversion of broilers. Curcumin and Probiotic increased PCV, RBCs, WBCs and lymphocytes count which reflected on broiler immune status consequently vaccinal reaction. Cost effectiveness study should be recommended.

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