RESPONSE OF BROILER CHICKENS FED DIETS SUPPLEMENTED WITH LACTOBACILLUS PLANTARUM AND ENTEROCOCCUS FAECIUM AS PROBIOTICS.

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ABSTRACT


Two feeding trails was conducted, using 336 day old broiler chicks to determine the response of broiler chickens fed diets supplemented with Lactobacillus plantarum and Enterococcus faecium, at the inclusion rate of 0, 0.05, 0.10 and 0.15 g. The studies lasted for 42 days. Experiment I determined the effect of two probiotic type while Experiment II considered different combinations of L. plantarum and E. faecium probiotics. Experimental design was 2 x 4 factorial in a completely randomized design (CRD) for Experiment I and CRD for Experiment II. Each of the experiments had seven treatments with each treatment replicated three times. Growth and performance, carcass and organ characteristics, haematology, serum biochemistry and feed cost, were measured. In Experiment-I, birds fed with starter broiler diet supplemented with 0.10g of E. faecium had significant (p<0.05) high final live weight (714.67g), daily feed intake (57.27g), daily protein intake (12.15g), high dressed percentage (48.93%), kidney (0.85%), crop (0.97%), small intestine (3.65%), large intestine (0.79%), high pack cell volume (32%), haemoglobin (9.50), red blood cell (5.20) and white blood cell (2.30). Starter diet supplemented with probiotics significantly (p>0.05) reduced urea (28.65), alkaline phosphatase (76.00), aspartate amino transferase (48.33), alanine amino transferase (27.67), cholesterol (48.57), low density lipoprotein (11.78) and the very low density lipoprotein (0.90). Diets supplemented with L. plantarum and E. faecium significantly (p<0.05) improved the final live weight (1876.67g) and daily weight gain (52.35g) of finisher broilers, with significantly low mortality rate (0.24%). The dressed percentage (65.08%), of finisher broilers fed diets supplemented with probiotics were significantly (p<0.05) improved. The haemoglobin (6.97g//dl), RBC (4.03ml/mm3) and WBC (0.44x103/µL) of finisher broilers fed diets containing probiotics were significantly lower than those fed with control diet. The total protein (5.46), was significantly (p<0.05) high, the creatinine (1.28), ALP (72.00), AST (19.33), cholesterol (52.47), LDL (13.09) and VLDLC (5.99) were significantly low. Birds fed diets supplemented with L. plantarum had significant (p<0.05) high total feed cost (₦491.14) and gross margin (₦878.85). In experiment-‍II, starter broilers dressed percentage (53.97%), crop (1.04), the small (5.37) and large (1.01) intestines, the PVC (32%), RBC (5.48ml/mm3) and WBC (2.10x103/µL) were significantly (p<0.05) high. While the FCR (1.95), cholesterol (81.37), triglyceride (81.37), LDL (4.87) and VLDLC (8.71) were significantly (p>0.05) low. The final live weight (1866.67g) and dressed percentage (62.66%), PVC (47%), RBC (6.85ml/mm3) and WBC (3.19x103/µL) of finisher broilers fed diets supplemented with probiotic combination of L. plantarum and E. faecium at varying inclusion levels were significantly (p<0.05) high. While the urea (19.37), creatine (1.01), AST (26.67), cholesterol (46.56), LDL (7.67) and VLDLC (23.74) were significantly (p>0.05) low, the feed cost (₦516.86) and gross margin (₦901.44) were significantly (p<0.05) high. Probiotics (L. plantarum and E. faecium) had positive impact on the growth performance, carcass and organ characteristics, haematology and serum biochemistry of broiler chickens.






TABLE OF CONTENTS

Title page                                                                                                                    i

Declaration                                                                                                                 ii

Certification                                                                                                               iii

Dedication                                                                                                                  iv

Acknowledgement                                                                                                      v

Table of contents                                                                                                        vi

List of tables                                                                                                               x

List of figures                                                                                                             xiv

Abstract                                                                                                                      xv


1.0       CHAPTER 1: INTRODUCTION

o         Background Information                                                                          1

o         Statement of Problem                                                                               2

1.3      Objectives                                                                                                        3

1.4       Justification                                                                                                    4

 

2.0       CHAPTER 2: LITERATURE REVIEW

2.1      Probiotics                                                                                                         6

2.2      Probiotics Micro-organisms                                                                            7

2.3      History of Probiotics                                                                                       8

2.4      Good Probiotics and their Benefits                                                                 9

2.5       Principles in Selecting Probiotic Strain for Animals                                     11

2.6      Probiotic Mode of Actions                                                                              13

2.6.1   Therapeutic action of probiotics in animals                                                    14

2.7.0    Classification of probiotics                                                                             15

2.7.1    The yeasts                                                                                                       15

2.7.2    The bacillus spores                                                                                         15

2.7.3    The lactic acid bacteria                                                                                  16

2.7.3.1 Lactobacillus species                                                                                      17

2.7.3.2 Specific roles of lactobacillus spp in animal nutrition                                   18

2.7.3.3 Lactobacillus plantarum                                                                                 19

2.7.4    Enterococcus species                                                                                      20

2.8       Rationale for Combination of two or more Probiotic Strain or Species            20

2.9       Application of Probiotics in Animal Nutrition                                               22

2.9.1    Effect of probiotics on laying hens                                                                 22

2.9.2    Effects of probiotics on broiler performance                                                 23

 

3.0       CHAPTER 3:  MATERIALS AND METHODS

3.1       Experimental Site                                                                                           25

3.2       Source of Probiotics                                                                                       25

3.3       Birds and their Management                                                                          25

3.4      Experiment – I                                                                                                 26

3.4.1    Experimental ‍‍ diet                                                                                          28

3.4.2    Experimental ‍‍ design and data collection                                                      28

3.4.3    Chemical and data analysis                                                                            30

3.5       Experiment – II                                                                                               30

3.5.1    Experimental ‍‍ diet                                                                                          31

3.5.2    Experimental ‍‍ design and data collection                                                      32

3.5.3    Chemical and data analysis                                                                            32

4.0       CHAPTER 4: RESULTS AND DISCUSSION                             

4.2.1    Growth and Performance                                                                               85

4.2.2    Carcass and organ yield                                                                                  88

4.2.3    Internal organs                                                                                                89

4.2.4    Haematology                                                                                                  89

4.2.5    Serum Biochemistry                                                                                       90

4.2.6    Feed cost                                                                                                         92

           

5.0       CHAPTER 5

5.1       Summary                                                                                                        93

5.2       Conclusion                                                                                                      94

5.3       Recommendation                                                                                           94                               

References                                                                                                     95                                           

 

 




 

LIST OF TABLES


3.1       Percentage composition of diets                                                                     27

4.1       Nutrient proximate composition of diet                                                         32

4.2       Effect of type of probiotic on growth and performance of starter broilers            33

4.3       Effect of inclusion level of probiotics on starter broiler                                34

4.4       Effect of interaction between type of probiotics and inclusion level on the

performance of starter broiler chickens.                                                         35

4.5       Effect of type of probiotic on the carcass yield of starter broilers                        36

4.6       Effect of inclusion level of probiotics on the carcass of starter broiler 37

4.7       Effect of interaction between type of probiotics and inclusion level on the

carcass of starter broilers.                                                                               39

4.8       Effect of type of probiotic on growth and performance of starter broilers            40

4.9       Effect of inclusion level of probiotics on the organs of starter broiler 41

4.10     Effect of interaction between type of probiotics and inclusion level on the

organs of starter broilers.                                                                                43

4.11     Effect of type of probiotic on the haematology of starter broilers                        44

4.12     Effect of inclusion level of probiotics on the haematology of starter broiler  45

4.13     Effect of interaction between type of probiotics and inclusion level on the

haematology of starter broilers.                                                                      46

4.14     Effect of type of probiotic on the serum profile of starter broilers                        47

4.15     Effect of inclusion level of probiotics on the serum of starter broilers            49

4.16     Effect of interaction between type of probiotics and inclusion level on

the serum profile of starter broilers.                                                               51

4.17     effect of type of probiotic on the growth and performance of finisher

broilers                                                                                                            52

4.18     Effect of inclusion level of probiotics on the performance of finisher

broilers                                                                                                            53

4.19     Effect of interaction between type of probiotics and inclusion level

on the performance of finisher broilers.                                                         55

4.20     Effect of type of probiotic on the carcass yield of finisher broilers                        56

4.21     Effect of inclusion level of probiotics on the carcass of finisher broilers            57

4.22     Effect of interaction between type of probiotics and inclusion level on the

carcass of finisher broilers.                                                                             59

4.23     Effect of type of probiotic on the internal organs of finisher broilers            60

4.24     Effect of inclusion level of probiotics on the organs of finisher broilers            62

4.25     Effect of interaction between type of probiotics and inclusion level on the

organs of finisher broilers.                                                                              64

4.26     Effect of type of probiotic on the haematology of finisher broilers                        65

4.27     Effect of inclusion level of probiotics on the haematology of finisher

broilers                                                                                                            66

4.28     Effect of interaction between type of probiotics and inclusion level

on the haematology of finisher broilers.                                                         67

4.29     Effect of type of probiotic on the serum profile of finisher broilers                        69

4.30     Effect of inclusion level of probiotics on the serum profile of finisher

broilers                                                                                                            70

4.31     Effect of interaction between type of probiotics and inclusion level on the

serum profile of finisher broilers.                                                                   72

4.32     Feed cost of broiler chickens fed diet supplemented with probiotics.       73

4.33     Growth and performance of starter broilers fed diet supplemented with

probiotics.                                                                                                       74

4.34     Carcass yield of starter broilers fed probiotic supplemented diet.                        75

4.35     Internal organs of starter broilers fed diet supplemented with probiotic.        76

4.36     Haematology of starter broilers fed diet supplemented with probiotics        77

4.37     Serum biochemistry of starter broilers fed diet supplemented with

probiotics.                                                                                                       78

4.38      Growth and performance of finisher broilers fed diet supplemented with

probiotics                                                                                                         79

4.39     Carcass yield of finisher broilers fed probiotic supplemented diets.               80

4.40     Internal organs of finisher broilers fed diet supplemented

with probiotics.                                                                                               81

4.41     Haematology of finisher broilers fed probiotic supplemented diet.                        82

4.42     Serum profile of finisher broilers fed diet supplemented with probiotics.       83

4.43     Feed cost of broiler chickens fed diet supplemented with probiotics.       84

 






 

LIST OF FIGURES


2.1       Schematic representation of probiotic concept                                              9

2.2       Diagram for probiotic selection in the poultry industry                                 14

 

 


 

 

 


CHAPTER 1

INTRODUCTION


1.1 BACKGROUND INFORMATION

Feed additives, improves the health, nutrition and general performance of animals. Feed is always a major expense in animal production and must be utilized as efficient as possible, ensuring the optimization of nutrient uptake, giving the best values. Feed additives are materials administered to animals mainly through feed to enhance the effectiveness of nutrients, which exert their effects in the gut or on the gut wall cells (McDonald et al., 2011). Primarily, feed additives include: antibiotics, prebiotics, probiotics, enzymes, organic acids, spray-dried plasma, phytogenics and modifiers of rumen fermentation.

Probiotics are cultures of beneficial bacteria from the healthy gut microflora that improve the balance of the intestinal milieu by modifying the intestinal microflora and suppressing enhanced inflammatory responses (Simmering and Blaut, 2001). Probiotics, or direct fed microbials, are naturally occurring live microbes added to feed to improve the gut flora (Fuller, 1989; Heyman and Ménard, 2002 and Biradar et al., 2004) of pigs, poultry and other animals. According to the definition by FAO/WHO (2001), probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Beneficial micro-organisms displace pathogenic micro-organisms and produce enzymes that complement the digestive ability of the host (McDonald et al., 2011). They are made-up of individual species or mixtures of lactic acid bacteria (LAB), yeasts or their end products which can have positive influence on digestion and the immune system (Anukam and Reid, 2008). Examples include Lactobacillus spp., Enterococcus spp., Bifidobacterium spp., Streptococcus spp., Lactococcus spp. among others (Hayashi et al., 2005; McDonald et al., 2011).

The mechanisms or mode of action of direct fed microbials (DFM) like probiotics, includes competitive exclusion (Fuller, 1989; Sissons, 1989; Bomba et al., 2002), aggregation of probiotics and pathogenic bacteria (Jiri and Christophe, 2015), reduced production of toxic amines (Broz and Paulus, 2015), nutrient synthesis and balancing of the intestinal pH (Bomba et al., 2002; Guerra et al., 1997 and Jiri and Christophe, 2015), nutrient absorption (Zhu, 2011), improves the intestinal immune system (Broz and Paulus, 2015) and total inhibition of pathogen’s growth (Bomba et al., 2002; Guerra et al., 1997; Mantere, 1995; Marinho et al., 2007).

 

1.2 STATEMENT OF PROBLEM

There is an increasing concern for the public health about the consequences of the long and increased use of antibiotics in livestock production. Thus, consideration must be given to the aftermath effects of its indiscriminate use which include antibiotic resistance, resulting from the ability of bacterial population to survive the effect of inhibitory concentration of antimicrobial agents (Catry et al., 2003) and accumulation of antibiotic residues in tissues of poultry birds that upon consumption poses risk to human health.

The use of antibiotics in animal feed as growth promoters has been completely banned by the European Union and most developed countries since 2006, based on their possible negative effects on human and animal health (EC Regulation No. 1831/2003; Vasileios et al., 2014).The removal of antibiotic growth promoters has led to animal performance problems and a rise in the incidence of certain animal diseases (Casewell et al., 2003 and WHO, 2002).

Animal production in recent days has been trapped between consumer’s concern on animal, increasing demand for animal products and concerns on risks for public health (Papatsiros et al., 2012). There is also, increasing public awareness about the risk of developing cross-resistance of pathogens to antibiotics (Mathew et al., 2007; Hunter et al., 2010). The total ban of antibiotic growth promoters in animal feed by most developed countries due to increased concern about the potentials for developing antibiotic resistant strains of bacteria within the food chain (Doyle, 2001), has led to an increased development and research on alternatives to antibiotics for use as feed additives in animal production. Most alternatives to antibiotics are inconsistent in their mode of action. This, calls for more research into safe and eco-friendly alternatives, which may include careful combination of different types and strains of probiotics like L. plantarum and E. faecium.


1.3 OBJECTIVES

The broad objective of the study was to determine the response of broiler chickens fed diets supplemented with Lactobacillus plantarum and Enterococcus faecium as probiotics. The specific objectives were to determine the:

a.     Growth performance of broiler chickens fed diets supplemented with L. plantarum and E. faecium.

b.     Carcass characteristics of broilers fed diets supplemented with L. plantarum and E. faecium.

c.     Hematology and serum biochemical composition of broiler chickens fed diets supplemented with L. plantarum and E. faecium.

d.     Feed cost of broiler chickens fed diets supplemented with L. plantarum and E. faecium.


1.4 JUSTIFICATION

With rising feed costs, animal producers and feed mills are looking for ways to optimize performance and increase the digestibility of nutrients. Brötel (2015) stated that probiotics helped farmers to make profit while at the same time reduce nutrient emission into the environment. Improved digestibility increases feed conversion ratio (FCR), resulting in reduction of feed cost to 4% (Brötel, 2015). Duncan et al. (2004) reported that lactic acid produced in vitro by lactic acid bacteria is used strictly by anaerobic butyrate producing bacteria of clostridial clusters IV and XIV for the production of large concentrations of butyric acid. This cross-feeding mechanism suggested that administration of lactic acid bacteria to farm animals has beneficial effect on performance. The intestinal microbiota has a specific multi-factorial ‘barrier’ impact, such as: induction of anatomical and physiological changes in the intestinal cell wall structure, immunological modifications in the gut and enhancement of the bird’s resistance to entero-pathogenic bacteria, such as C. perfringens (La Ragione et al., 2004; Hofacre et al., 1998; Ng et al., 2009).

Delia et al. (2012) outlined the beneficial effect probiotics to include resistance to infectious diseases, increased growth rate, improved feed conversion, improved digestion, better nutrient absorption, provision of essential nutrients, increased egg production, improved egg quality, and improved carcass quality and less contamination. Samli et al.(2007) observed that supplementation of E. faecium in a broiler diet increased the population of lactic acid bacteria in broiler droppings and increased villus height in the ileum, thereby enhancing broiler’s performance with respect to weight gain and feed conversion ratio. The inclusion of L. plantarum, L. acidophilus and E. faecium at 1%, 3% and 5% of a broiler diet significantly improved the amino acid level of the breast and leg muscles but with more difference in the leg (Abdulwahab and Horniakova, 2013). Saurabh et al, (2013) observed that at low inclusion level, E. faecium acts effectively as a probiotics with negative correlation in biological performance at high inclusion levels.

Combination of L. plantarum and E. faecium in broiler diets is expected to synergize to enhance better nutrient utilization, feed efficiency, weight gain, healthy gut and immunity with very low pathogenic bacteria population and low serum lipid profile. Therefore, there is need to determine probiotic strain combination and the inclusion level that will enhance the performance of broiler chickens in the humid tropics.

 

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