EFFECT OF PROBIOTIC SUPPLEMENTATION ON SOME BLOOD PARAMETERS OF FINISHER BROILER CHICKENS

Transcription

1 EFFECT OF PROBIOTIC SUPPLEMENTATION ON SOME BLOOD PARAMETERS OF FINISHER BROILER CHICKENS Okey 1, S. N., Ndelekwute 2, E. K., Okey 1, O. N., Marire 1, B. N. and Omeke 1, B. C. O. ABSTRACT 1 Department of Animal Health and Production Faculty of Veterinary Medicine, University of Nigeria, Nsukka. 2 Department of Animal Science, University of Uyo, Nigeria. nnamuokey@yahoo.com Tel: An experiment was carried out to determine the dietary effect of Saccharomyces cerevisiae (SC), Lactobacillus acidophilus (LA) and Lactobacillus sporogenes (LS) as probiotics on hematological and serum biochemical parameters of finisher broiler chickens. One hundred and twenty unsexed day old Anak strain of broilers were used. They were maintained ad- libitum for 4 weeks on a starter diet containing no probiotics. At the end of which, the birds were distributed into four treatment groups (T1- T4) of thirty birds each. Each treatment was replicated three times each having 10 birds. The experiment was arranged in completely randomized design (CRD). T1 was the control and fed diet containing no probiotics. T2-T4 was fed diets which contained 0.8g kg -1, 0.5g kg -1 and 0.5g kg -1 SC, LA and LS respectively. They were fed diet and water ad libitum for 5 weeks. At the beginning of the experiment which was at the 5 th week and at the end of the experiment at the 8 th week, blood samples were collected and analyzed. Changes in hematological parameters were not significant (P>0.05) except the red blood cells counts and packed cell volume where feeding of SC resulted in greater number of red blood cells compared to the control (P<0.05). Total protein, albumin and globulin levels were not affected by any of the probiotics. Cholesterol level was significantly lowered by the probiotics. Therefore any of the probiotics could be used in finisher broiler diets without detrimental effect to the health especially SC which increased the red blood cells. INTRODUCTION The poultry industry has always been confronted with a lot of challenges. Among these are economic losses due to infectious diseases caused by viruses, bacteria, fungi, protozoa, and the cost of medication. This has led to increased use of antibiotics in poultry farming for therapeutic and prophylactic purposes. The presence of antibiotic residues in poultry meat and eggs has been reported to have harmful effects on human consumers (Dibner and Richards, 2005). The residues of antibiotics can cause resistance of human flora and pathogenic microbes to antibiotics. Eden (2003) mentioned that with increasing concerns about antibiotic resistance, and the ban on sub-therapeutic antibiotics usage in Europe and United States, there is increasing interest in finding alternative to antibiotics for poultry production such as probiotics. Fuller (1989) defined probiotics as a live microbial feed supplement which beneficially affects the host by improving its microbial balance and promoting performance. According to Food and Agriculture Organization (FAO, 2001) probiotics are live microorganisms which when consumed in adequate amount confers a health benefit on the host. The microorganisms commonly used in animal feed are mainly gram-positive bacteria strains belonging to the genera of Lactobacillus, Enterococcus, Streptococcus, Bfidobacterium, Pediococus and and fungal species such as Sacchromyces cerevisiae (Starvic et al., 1995). The most important advantage of probiotics is that they do not have residues in animals. Also, their ability to increase growth and promote health are claimed to be due to their competitive exclusion of pathogenic bacteria (Balcazar, 2003) and as a source of nutrients and its enzymatic contribution to digestion (Sakata, 1990). To combat poultry diseases and maintain profitable production, early diagnosis of diseases is necessary, and an assessment of the hematological parameters usually provides valuable information essential for this (Campbell and Coles 1986; Dein 1986; Koinarski et al 2001). In veterinary practice, a diagnosis is considered incomplete if information from history and clinical examination is not combined with laboratory test results, including results of assessment of some hematological and serum biochemical indices (Carter 1996; Ugochukwu 2001). Evaluation of hematological parameters usually supplies the veterinarian with information that will enable him decide whether an animal is anaemic, normal, polythemic and could point to the possible cause of the abnormality (Campbell and Coles 1986; Dein, 1986; Ihedioha and Chineme, 2004). Gheisari and Kholeghipour (2006) reported that the use of probiotics such as live yeast had no significant effects on hematological indices like RBC, WBC and PCV. On the contrary, Onifade (1997) reported a positive correlation between dietary level of Sacchromyces cerevisaie with hematological indices like RBC, WBC, and PCV in rabbit and broiler chickens. Gheisari et al. (2006) stated that serum concentration of albumin and total protein are not affected by live yeast supplemented in broiler feed. Cholesterol measurements are used in the diagnosis and treatments of lipid lipoprotein disorders. It has been reported that probiotics could possess the property of reducing cholesterol in the blood, which is due to the inhibition of hepatic synthesis and their capacity of deconjugating the biliary salts (Mercenier et al., 2002; Lim et al., 2004). Onifade (1998) also stated that there is reduction in circulating cholesterol when yeast is supplemented NJAFE VOL. 11 No. 2,

2 in broiler feed. On the contrary Djouvinov et al. (2005) did not observe any variations of cholesterol and triglycerides contents in chicken s blood, while using mixture of different strains of probiotics. MATERIALS AND METHODS The experiment was carried out at the Teaching and Research farm of University of Nigeria Nsukka, in the humid rain forest zone of Nigeria. The probiotics used in this study were Lactobcillus acidophilus (45 Million CFU), Lactobacillus sporogenes (50 million CFU) and yeast (Sacchromyces cerevisae). The quality of the yeast (Sacchromyces cerevisiae) was determined by culturing on Sabouraud Dextrose Agar for 48 hours and the number of yeast cells per gram was counted to be 48 Million CFU using a digital colony counter. Experimental diets A basal starter and finisher diets were formulated to conform to the nutrient requirements of broilers in the tropics according to NRC (1994) as shown in Table. 1. Table I: Composition of experimental diets Ingredients (%) Starter Finisher Maize Soya Bean Meal Palm Kernel Cake Fish Meal Bone Meal Salt (Na cl) Lysine Methionine Premix Calculated Nutrient Composition (%) Crude Protein (%) Metabolizable Energy (Kcal/kg) Calcium (%) Phosphorus (%) Lysine (%) Methionine (%) Experimental design A total of 120 day old Anak broiler chicks were used. They were divided into four treatment groups (T1-T4) of 30 birds each. Each treatment had 3 replicates of 10 birds each arranged in a completely randomized design (CRD). At the starter phase, all the treatments were offered a single starter diet for 4 weeks. At the end of the starter phase, each treatment group was assigned a diet which was formed by adding to a basal finisher diet 0.8g of Sacchromyces cerevisiae, 0.5g Lactobacillus acidophilus, and 0.5g of Lactobacillus sporogenes per kg of the basal diet representing T2, T3 and T4 respectively. Treatment 1 was maintained on the basal diet and served as control. The inclusions of bacteria probiotics were as recommended by the manufacturers, while Sacchromyces cerevisiae was at its best level of inclusion as recommended by Ezema (2007). All the birds were fed and watered ad-libitum till the end of the experimental period which lasted for 4 weeks. Appropriate vaccinations and preventive medications were carried out. Management of birds The birds were brooded for three weeks on a single formulated starter diet. Vaccines were administered against Newcastle and Gumboro diseases. At the end of the brooding, the birds were grouped into treatments and transferred to open sided rearing house where they were reared on a deep litter floor till the end of the experiment. Data collection At the beginning and end of the experiment, five birds were randomly selected from each replicate. Blood samples were collected from the wing vein of each bird. The blood samples were collected into McCartney bottles containing ethylenediaminetetra acetic acid for determination of hematological parameters. Blood samples for serum biochemical analysis were allowed to clot in the container. The blood samples were analyzed based on the methods of Decie and Lewis (1995) and Coles (1986). Statistical analysis At the end of the experiment, the data obtained were subjected to one-way statistical analysis of variance according to Steel and Torrie (1980) and treatment means compared using Duncan s Multiple Range Test (Duncan, 1995). NJAFE VOL. 11 No. 2,

3 RESULTS AND DISCUSSION The values of hematological and serum biochemical parameters at the beginning of the experiment (i.e. the end of the starter phase) are shown in Tables 2 and 4 respectively. All the values were statistically similar. The values of hematological indices (Table 3) at the end of the experiment at the finisher phase, indicated that the probiotics did not significantly (P>0.05) influence hemoglobin, white blood cells, lymphocyte, neutrophil and monocyte. However, significant (P<0.05) effect occurred in red blood cells and packed cell volume. All the probiotics brought about increase in red blood cells and packed cell volume compared to the control. C. cerevisiae and L. sporogenes produced higher red blood cells than L. acidophilus. The packed cell volume was higher in birds that consumed C. cerevisiae than the birds that consumed the Lactobacillus spp. This result was in line with Onifade (1997) who observed higher red blood cells and packed cell volume in broiler birds fed these probiotics. The increase in red blood cells and packed cell volume brought about by the feeding of the probiotics to the birds is an indication that they could be used to improve the health status of broiler chickens. The effect of the probiotics on serum biochemical parameters is indicated in Table 5.There were no significant (P>0.05) differences in all the indices measured except cholesterol. The level of the cholesterol was significantly reduced by all the probiotics. There were no significant (P>0.05) differences between the cholesterol levels produced by the probiotics. These findings are in agreement with those of Dimcho et al. (2005) who found that probiotic supplementation did not significantly affect the total protein concentration of chickens. The work done by Gheisari et al. (2006) confirmed that serum concentration of albumin, globulin and total proteins were not affected by probiotic supplementation in broiler feed. In addition, Kanashiro et al. (2001) and Djourinov et al. (2005) did not observe any significant variations in cholesterol levels among broilers fed a mixture of different strains of probiotics. Onifade (1998) stated that there was reduction in circulating cholesterol in the blood when probiotics such as yeast is supplemented in broiler feed. This could probably account for the reduction in the cholesterol level in the supplemented groups when compared with the control group. In conclusion, the probiotics used in this study especially Sacchromyces cerevisiae had a significant improvement in the hematological values (Red blood cells and Packed cell volume) in the broilers. This could probably be due to high nutritive value of yeast resulting from the presence of protein, vitamins and enzymes in yeast cell (Erwin, 1953) and hence could be recommended to be incorporated in diets for broiler chickens. Table 2: Hematological Parameters of starter broiler chicks at the beginning of the experiment Parameters T1 T2 T3 T4 Mean RBC count (10 6 µl -1 ) 3.33 ± ± ± ± 0.12 Mean PCV (%) ± ± ± ± 0.65 Mean Hemoglobin concentration (g dl -1 ) 8.85 ± ± ± ± 0.24 Mean WBC count (x 10 3 µl -1 ) ± ± ± ± 0.23 Mean Lymphocyte count (x10 3 µl -1 ) 8.16 ± ± ± ± 0.22 Mean Neutrophil count (x10 3 µl -1 ) 3.28 ± ± ± ± 0.10 Mean Monocyte count (x10 3 µl -1 ) 0.15 ± ± ± ± 0.09 ±SD, RBC = red blood cells, WBC = White blood cells Table 3: Effect of probiotics on hematological parameters ±SD of finisher broiler chickens Group Parameter T1 T2 T3 T4 Mean RBC count (10 6 µl -1 ) 2.71 ± 0.41 c 3.86 ± 0.15 b 3.04 ± 0.36 ab 3.71 ± 0.22 a Mean PCV (%) ± 0.85 a ± 1.08 b ± 1.08 ab ± 1.08 ab Mean hemoglobin concentration (g dl -1 ) 9.05 ± ± ± ± 0.35 Mean WBC count ( x10 3 µl -1 ) ± ± ± ± 1.47 Mean lymphocyte count (x10 3 µl -1 ) ± ± ± ± 0.11 Mean neutrophil count (x10 3 µl -1 ) 4.20 ± ± ± ± 0.21 Mean monocyte count x10 3 µl -1 ) 0.19 ± ± ± ± 0.03 abc Means along the row with different superscripts are significant (P < 0.05) different. RBC = red blood cells, WBC = White blood cells NJAFE VOL. 11 No. 2,

4 Table 4: Serum Biochemical parameters ±SD of starter broiler chicks at the beginning of the experiment Parameter T1 T2 T3 T4 Total Protein ( g dl -1 ) 4.60 ± ± ± ± 0.28 Albumin ( g dl -1 ) 1.70 ± ± ± ± 0.09 Globulin ( g dl -1 ) 2.90 ± ± ± ± 0.22 Cholesterol ( g dl -1 ) ± ± ± ± 6.66 Table 5: Effect of probiotics on Serum biochemical parameters ±SD of finisher broiler chickens Parameters T1 T2 T3 T4 Total Protein ( g dl -1 ) 4.70 ± ± ± ± 0.41 Albumin ( g dl -1 ) 1.87 ± ± ± ± 0.25 Globulin ( g dl -1 ) 2.83 ± ± ± ± 0.18 Cholesterol ( g dl -1 ) ± 6.39 a ± b ± 5.20 b ± 5.20 b abc Means along the row with different superscripts are significant (P < 0.05) different REFERENCES Balcazar, J. I Evaluation of probiotic bacterial strains in Litopenaeus vannamei. National Centre for Marine and Aquaculture Research Report. Guayaquil, Ecuador. Campbell, T. W. and Cole E. H Avian Clinical Pathology. In: Coles, E. H. (ed.). Veterinary Clinical Pathology, 4 th ed. W. B Sanders Company, Philadelphia. Pp: Carter, G. R Veterinarians guide to laboratory diagnosis of infectious diseases. Veterinary Medicine Publishing Company. Kansas, pp Coles E. H Veterinary Clinical Pathology. 4 th edition. WB Saunders Company, Philadelphia, London, Toronto: Dacie, J. V. and Lewis, S. M Practical Hematology 8 th Edition. Churchill Livingstone Publication. United Kingdom. Dein, E. J Hematology. In: Harrison, G. J. and Harrison, L. R. (eds). Clinical Avian Medicine and Surgery. WB Sanders Company. Philadelphia, pp Dibner, J. J. and Richards, J. D Antibiotic growth promoters in agriculture: History and mode of action. Poultry Science 84: Dimcho, D., Svetlana, B., Tatiana V. and Tsvetomiva, S Effect of Feeding Lactina Probiotic on Performance, some blood parameters and caecal micro flora of mule ducklings. Trakia Journal of Sciences. 3: Djonvinov, D., Stefanor, M., Boichera, S. and Vlaikova, T Effect of diet formation, on basis of digestible amino acids and supplementation of probiotics on performance of broiler chicks. Trakia Journal of Sciences, 3(1): Duncan, D. B Multiple Range and Multiple F- Tests. Biometrics. 11: Eden F. W Alternative for antibiotics use in Poultry: Probiotics. British. Journal. Poultry. Science. 5(2): Erwin, S Food and Feed yeast in Germany. Unasylva. FAV Corporate Document. Vol.7, N0.4. Ezema, C Growth rate of broilers on palm kernel cake based diet supplemented with bioactive yeast. M.Sc. Thesis, University of Nigeria Nsukka. FAO, Evaluation of health and nutritional properties of probiotics in food including powder milk with live bacteria. Joint FAO/WHO Report. Cordoba, Argentina. Fuller, R Probiotics in man and animal. Journal of Applied Bacteriology. 66: Gheisari, A. A. and Kholeghipour, B Effect of dietary inclusion of live yeast (Sacchromyces cerevisiae) on growth performance, immune responses and blood parameters of broiler chickens. EPC-XII European Poultry Conference, Verona Italy, Pp: Ihedioha, J. I and Chineme, C. N Haematopoetic System. In: Ihedioha J. I. and Chineme C. N (eds). Fundamentals of Systemic Veterinary Pathology, Great AP Express Publishers, Nigeria, pp Kanashiro, A. M., Bittino, T. A., Ferreiara, F., De Castro, A.G. and Ferreiara, A. J Infuence of probiotic continuous administration to broiler on serum enzymes activities and serum cholesterol concentration. Arq. Inst. Bio., 68 (2): Koinarski, V., Angelov, G. and Lalev, M Hematologic and blood electrolyte changes in broiler chickens experimentally invaded with Eimeria tenella. Bulgeria. Journal of Veterinary Medicine. 4: NJAFE VOL. 11 No. 2,

5 Lim, H. L., Kim S. Y. and Lee, W. K Isolation of cholesterol-lowering lactic acid bacteria from human intestine for probiotic use. Journal. Of Veterinary. Science. 5(4): Mercenier, A. Pavan, S. and Pot, B Probiotics as bio therapeutic agents: Present knowledge and future prospects. Current. Pharmacology. Design. 8: National Research Council, (1994). Nutrient Requirement of Poultry. 7 th rev. ed. Washington D. C: National Academy Press. Onifade, A. A Growth performance, carcass characteristics, organ measurements and hematology of broilers chicken feed a high fibre diet supplemented with antibiotics or dietary yeast. Die Nahrung. 41: Onifade, A. A. and Abu, O. A Proposing fortification of foods with yeast for optimal nutrition value and salubrious effects. Nutrition and Food Science. 4: Sakata, T Microflora in the digestive track of fish and shell fish. In: Lessel, R. (ed), Microbiology in Peocilotherms, Elservier, Amsterdam Starvic, S. and Kornegay, E. T Microbial Probiotics for Pigs and Poultry, Biotechnology in animal feeds and animal feeding. R. J. Wallace, and A. Chesson, (eds.) VCH, New York. pp Steel, R. G. D. and Torrie, J. H Principle and Procedures of Statistics. 3 rd Edition. MC Graw-Hill Book Company, Inc. New York, USA. Pp Ugochukwu, E. I Laboratory examination in veterinary clinical diagnosis. In: Ugochukwu E.I. (ed). Manual of Veterinary Clinical Practice. Amazing Grace Printing and Publishing Co. Nigeria. pp NJAFE VOL. 11 No. 2,