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STUDIES ON THE PHYTOCHEMICAL, MEDICINAL AND NUTRITIONAL VALUES OF CONGRONEMA LATIFOLIA (UTAZI) IN RABBIT AND POULTRY PRODUCTION

ABSTRACT

Experiments were conducted to evaluate the phytochemical medicinal and nutritional values of Congronema latifolia (utazi) leaf meal (CLLM) in rabbit and poultry production. The proximate analysis of the leaf meal (CLLM) yielded 8.04%, 14.25%, 60.39%, 6.26%, 2.84% and 2.84% moisture, crude protein, NFE, ash, ether extract and crude fibre respectively on dry matter bases. The methanolic extract of CLLM exhibited a minimum inhibitory concentration (MIC) of 15.62 mg/ml against Pseudomonas aeruginosa and E. Coli while Staphylococcus aureus, Klebsiella spp and Salmonella spp were all inhibited at a concentration of 250mg/ml. The minimum bactericidal concentrations (MBC) of the above five organisms tested were 31.25, 31.25, 250, 250 and 250 mg/ml, respectively, while the diameters of the zone of inhibition measured at 250mg/ml of the leaf meal extract in centimeters were 1.4, 1.5, 0.0, 0.0 and 0.3 for Ps. aeruginosa, E. Coli, S. aureus, Klebsiella spp, and Salmonella spp, respectively. This result suggests that the methanolic extracts of CLLM are very effective against Ps. aeruginosa and E. coli. In the rabbit experiment, 4 experimental diets were formulated such that diet 1 (control) contained 0% CLLM, while diets 2, 3 and 4 contained 10%, 20% and 30% CLLM, respectively. Each diet was fed to a group of 9 grower rabbits for 49 days. There were no significant differences (P>0.05) among the treatment groups for average final body weight, average body weight gain and average daily feed intake. The feed conversion ratio was also not affected by the treatments (P>0.05). The dressing percentage and internal organs (percentage hearts, lungs and pancreas weights) were not significantly affected (P>0.05) by the treatments. Dietary levels of 20% and 30% CLLM significantly (P<0.05) increased the percentage weight of liver and kidneys. Haematological parameter which showed significant increase (P<0.05) was the mean cell haemoglobin (MCH), at 20% and 30% levels of CLLM, while the white blood cell concentration decreased significantly (P<0.05) at 10%, 20% and 30% levels. Other indices (Hb, RBC, ESR, PCV, MCV and MCHC) were similar (P>0.05). Dietary levels of 10%, 20% and 30% CLLM, signficantly (P<0.05) increased total serum protein, but other serum bio-chemical parameters were not affected by the treatments (P>0.05). There were no lesions of pathologic significance in the tissues (liver, kidney and pancreas) examined.

In the broiler experiment, 5 experimental diets were formulated such that diet 1 (control) contained 0% CLLM while diets 2, 3, 4 and 5 contained  2.5%, 5.0% 7.5 and 10.0% CLLM respectively. Each diet was fed to a group of 30 broilers (one week old) for 49 days. The average final body weight, average body weight gain and feed conversion ratio indicated no significant (P>0.05) treatment effect. There was significant depression in average daily feed intake (P<0.05) at 10% dietary level.  Ten percent dietary level also significantly (P<0.05) depressed RBC concentration but other haematological indices indicated no significant (P>0.05) treatment effect. Serum glucose was significantly (P<0.05) reduced at 7.5% and 10%; AST dropped significantly (P<0.05) at 5%, 7.5% and 10%; bilirubin increased significantly (P<0.05) at 5%, 7.5%, 10% and 2.5% dietary levels of CLLM. Others serum parameters were not affected (P>0.05) by the treatments. Dietary levels of 2.5%, 5.0% and 7.5% CLLM (P<0.05) increased dressing percentage of the experimental birds. Percentage weights of necks, wings, thighs, drum sticks, breasts, hearts, liver, spleen, lungs and pancreas (expressed as percentage of live weights) were similar (P>0.05). However, dietary levels of 2.5% – 10.0% CLLM yielded significantly higher (P<0.05) percentage weights of proventriculus than the control diet. There were no lesions of pathologic significance in the liver, kidney, proventriculus and pancreas. It would therefore seem that rabbits can tolerate dietary levels of up to 30% CLLM while 7.5% inclusion level of the leaf meal can support normal broiler production. 

Keywords: Poultry, Poultry Production, Phytochemical, Nutritional Values

TABLE OF CONTENTS

Title Page                                                                                                     i

Certification                                                                                                    iii

Dedication                                                                                                       iv

Acknowledgement                                                                                         v

Abstract                                                                                                        viii

Table of contents                                                                                               x

List of tables                                                                                                  xix

List of figures                                                                                               xxii

CHAPTER ONE

1.0Introduction                                                                                              1

1.1Research Objectives                                                                                  4

CHAPTER TWO

2.0Literature review                                                                                       6

2.1Chemical composition of leaf meals                                                          6

2.1.1 Fibre content of leaf meals                                                                     7

2.1.2 Essential amino acid composition of leaf meals                                      7

2.1.3 Mineral composition of leaf meals                                                         8

2.1.4 Metabolic energy content of leaf meals                                                  12

2.1.5 Carotenoids content of leaf meals                                                           12

2.2Nutritive value of leaf meals                                                                      15

2.3Leaf meals in non-ruminant nutrition                                                         16

2.3.1 Leucaena  leucocephala leaf meal                                                           18

2.3.2 Gliricidia sepium                                                                                   21

2.3.3 Sesbania species leaf meal                                                                     21

2.3.4 Microdesmis puberula leaf meal                                                             21

2.3.5 Cajanus cajan leaf meal                                                                          22

2.3.6 Vernonia amygdlina leaf meal                                                                23

2.3.7 Jacaranda mimosifolia                                                                            23

2.3.8 Cassava leaf meal                                                                                   24

2.4 Leaf meal as source of pigmenting xanthophylls                                        27

2.5 Egg yolk pigmentation                                                                              30

2.6 Sources of carotenoids                                                                              32

2.7 Limitations to leaf meal utilization by monogastrics                                   33

2.7.1 Anti-nutritional factors in leaf meal                                                        35

2.7.1.1 Protease inhibitors                                                                               36

2.7.1.2. Lectins and their nutritional significance                                             37

2.7.1.3 Amylase inhibitors                                                                             40

2.7.1.4 . Cyanogenic glycosides in plants

and their biochemical properties                                                         40

2.7.1.5 Oligosaccharides                                                                                 43

2.7.1.6 Saponins                                                                                            44

2.7.1.7 Phenolics                                                                                           44

2.7.1.8 Phytate and its nutritional significance                                                47

2.7.1.9Oxalate and its nutritional significance                                                 48

2.7.1.10Alkaloids and their nutritional significance                 49

2.8 Ethno-veterinary potential of leaf meals

CHAPTER THREE

           50

3.0Materials and methods             57

3.1Location of study               57

3.2Preparation of Congronema latifolia (utazi) leaf meal (CLLM)

3.3Proximate analysis and metabolisable energy determination

          57

       of CLLM                         58

3.4Mineral analysis                             58

3.5Amino acid analysis                               59

3.5.1. De-fatting of sample                          59

3.5.2  Nitrogen determination                            59

3.5.3  Hydrolysis of sample                         60

3.5.4 Loading of the hydrolysate into the TSM Analyzer                       61

3.6Phytochemical screening of C. latifolia leaf meal                             61

3.6.1 Determination of alkaloids                         61

3.6.2 Determination of flavonoids                             62

3.6.3 Determination of tannins                          63

3.6.4 Determination of saponin                          64

3.6.5 Determination of Phenols                          65

3.7 Bio-activity studies of CLLM              66

3.7.1Preparation of nutrients broth               67

3.7.2Minimum inhibitory concentration (MIC) test              67

3.7.3Minimum bactericidal concentration test                                                 68

3.7.4Zone of inhibition test                                                                            68

3.8Feeding trial with rabbits                                                                           69

3.8.1Experimental diets                                                                                   69

3.8.2Experimental rabbits and design                                                              69

3.8.3Data collection                                                                                        70

3.8.4 Haematological studies                                                                           70

3.8.4.1 Estimation of haemoglobin concentration                                             71

3.8.4.2 Red blood cell (erythrocyte) count                                                       73

3.8.4.3 White blood cell (leucocyte) count                                                       74

3.8.4.4 Erythrocyte sedimentation rate                                                             75

3.8.4.5 Estimation of packed cell volume (PCV)                                              75

3.8.4.6 Mean cell volume (MCV)                                                                    76

3.8.4.7 Mean cell haemoglobin (MCH)                                                           76

3.8.4.8 Mean cell haemoglobin concentration (MCHC)                                    77

3.8.5Serum biochemistry                                                                                77

3.8.5.1 Estimation of total serum protein                                                         78

3.8.5.2. Serum albumen/globulin assay                                                            79

3.8.5.3 Creatinine assay                                                                                  80

3.8.5.4. Estimation of serum total cholesterol                                                  81

3.8.5.5 Determination of urea                                                                         83

3.8.5.6 Determination of glucose                                                                    84

3.8.5.7 Determination of serum calcium                                                         85

3.8.5.8 Serum sodium                                                                                    86

3.8.5.9 Serum potassium determination                                                          87

3.8.5.10 Aspartate aminotransferase                                                               88

3.8.5.11 ALT determination                                                                            89

3.8.5.12. ALP determination                                                                           90

3.8.5.13Determination of bilirubin                                                                   91

3.9 Internal organs evaluation                                                                          92

3.10 Histopathological examination                                                                 93

3.11 Statistical analyses                                                                                  93

3.12 Feeding trial with boilers                                                                         94

3.12.1 Experimental diets                                                                                94

3.12.2 Experimental birds and design                                                              94

3.12.3 Data collection                                                                                     95

3.12.4 Hematological and serum biochemical studies                                       95

3.12.5 Carcass and organ weights evaluation                                                   96

3.12.6 Histopathological study                                                                        99

3.12.7 Statistical Analysis                                                                               99

CHAPTER FOUR.

4.0 Results and Discussions                                                                          100

4.1 Experiment 1: Determination of chemical composition

Amino acid profile, phytochemical composition and

Bio-activity studies of CLLM                                                                  100

4.1.1 Chemical composition of CLLM                                                          100

4.1.2 Amino acid composition          100

4.1.3 Phytochemical composition of CLLM            101

4.1.4. Bio–activity studies of CLLM         101

4.1.5. Experiment 2: Rabbit trial                        102

4.2.1. Average final body weights                       102

4.2.2. Average body weight gain                  109

4.2.3 Average daily feed intake                    109

4.2.4. Average daily body weight gain                        109

4.2.6 Haemathological indices of grower rabbits                      109

4.2.6.1 Haemoglobin concentration                    110

4.2.6.2. Red blood cell concentration                  110

4.2.6.3 White blood cell concentration                     110

4.2.6.4. Erythrocyte sedimentation rate                   110

4.2.6.5. Packed cell volume                  112

4.2.6.6. Mean cell volume                    112

4.2.6.7. Mean cell haemoglobin                   112

4.2.6.8. Mean cell haemoglobin concentration                  112

4.2.7 Serum biochemical indices of grower rabbits                  112

4.2.7.1. Total serum protein                       113

4.2.7.2. Serum albumen                      113

4.2.7.3. Serum globulin                       113

4.2.7.4. Cholesterol                      113

4.2.7.5. Creatinine                        113

4.2.7.6. Urea concentration                                                                 115

4.2.7.7. Glucose concentration                                                            115

4.2.7.8. Serum sodium                                                                        115

4.2.7.9. Serum calcium concentration                                                  115

4.2.7.10. Serum potassium                                                                  115

4.2.7.11. Aspartate transaminase (AST)                                                116

4.2.7.12. Alanine transaminase (ALT)                                                 116

4.2.7.13. Alkaline phosphatase (ALP)                                                 116

4.2.7.14. Total (unconjugated) bilirubin                                                116

4.2.7.15. Direct (conjugated) bilirubin                                                 116

4.2.8. Carcass and organ weight characteristics                                    117

4.2.9. Histopathological studies (Rabbit)                                             119

4.3 Experiment 3 (broiler trial)                                                            120

4.3.1. Average final body weights                                                       120

4.3.2. Average body weight gain                                                         120

4.3.3.  Average daily feed intake                                                         120

4.3.4. Average daily body weight gain                                                 120

4.3.5. Feed conversion ratio (FCR)                                                      123

4.3.6 Haematological indices of experimental broiler birds                  123

4.3.7 Serum biochemical indices of broiler birds                                 126

4.3.8 Carcass  and organ weights evaluation                                         129

4.3.9 Histopathological studies (Broilers)                                            133

4.4.1. Proximate composition of CLLM                                             134

4.4.2 Amino acid analysis                                                                   135

4.4.3. Phytochemical composition of CLLM                                       138

4.4.4 Bio-activity studies of CLLM                                                     139

4.4.5.  Rabbit trial                                                                               140

4.4.5.1. Body weight gain                                                                   140

4.4.5.2. Average daily feed intake                                                        141

4.4.5.3. Feed conversion Ratio                                                            142

4.4.5.4. Mortality                                                                                143

4.4.6. Carcass characteristics and organ weight                                    143

4.4.7. Histopathology of rabbits                                                           144

4.4.8. Haematology of grower rabbits                                                  145

4.4.9. Serum biochemistory of grower rabbits                                      147

4.4.10. Experiment 3 (Broiler Trial)                                                    150

4.4.10.1 Body weight gain                                                                  151

4.4.10.2 Average daily feed intake                                                       151

4.4.10.3 Feed conversion ration (FCR)                                                152

4.4.10.4 Body pigmentation                                                                153

4.4.10.5 Mortality of broilers                                                               153

4.4.10.6 Carcass characteristics                                                           153

4.4.11 Histopathology of broilers                                                       154

4.4.12 Haematology  of broilers                                                         155

4.4.13 Serum biochemistry of finisher broilers                  156 CHAPTER FIVE

5.0. Conclusion and recommendations                                                160

Experiment 1: Bio-activity studies of CLLM 160

5.1.1 Conclusions                                                                               160

5.1.2 Recommendations                                                                      160

Experiment 2: Rabbit tiral                               161

Conclusions                              161

Recommendations                              161

Experiment 3: Broiler trial                               162

Conclusions                              162

Recommendations                              163

Genera conclusion and recommendation                    163

General conclusions                               163

General recommendations                               163

References                                                                                        164

CHAPTER ONE

 INTRODUCTION

Livestock industry is of socio-economic and nutritional importance for nations worldwide. Apart from providing a means of livelihood for a significant population of the farming families, it contributes significantly to the Gross Domestic Product (GDP) of the nation (Duruna, 1996).

Worthy of note is the fact that the success of the industry hings basically on nutritional and health management of the animals. Over the years, poultry industry and swine production have been among the most lucrative sectors of agriculture in Nigeria. Indeed from inception, through the early 1980s, the industries have experienced a significant development and expansion. As a result, there was an appreciable national flock size and worthwhile income for farmers. This favourable trend was, however, short-lived. During the last two decades, the industry has witnessed a serious set back due largely to inadequate feed supply which stems from unavailability of protein concentrate and short supply of energy sources. Moreso, the problem has been aggravated by competitive demand for some of the existing feed ingredients like maize, millet and soybean for livestock feeding, and human consumption. This dismal trend has been responsible for the prohibitive cost of finished feeds.

Agreeably, feed cost accounts for 60 – 80% of the total production cost of commercial poultry and intensive livestock production in general. The high feed cost relative to monogastric livestock products from the early 1980s to date has drastically slashed farmer’s profit margin. The obvious implication here is that revenue accruing from the sales of pork, poultry meat, eggs, and other commercial livestock products cannot adequately compensate for the cost of production of the same. The consequence of this has been far-reaching. Resource-poor farmers have been forced out of production while “bigger farmers” have reduced stock population to as low as 30 to 50%, while new entrants have been demoralized. In certain cases, sub-standard feeding has been resorted to, leading to poor production. In view of this limiting circumstance, it would seem reasonably important to search for cheaper, readily available, nonconventional feed sources that will be well suited for a sustainable monogastric production industry. Such feed should of course be low in competitive demand for human food. It is this line of thought that has generated this research interest in Congronema latifolia, known in Efik, Ibibio and Igbo languages as utazi to determine its value as a feed ingredient for poultry and rabbit diets.

The use of leaf meal of plants as feed ingredients as an alternative to conventional feed sources is a novel area of research in animal nutrition. A number of workers (Tewe, 2003; Adegbola, 2004) have shown that various alternative feedstuffs have been fed to poultry with remarkable results. Some of these alternative sources including the leaf meals of some tropical legumes and browse plants, rich in nutrients like vitamins, minerals and oxycarotenoids have been reported (Vohra et al., 1972; Okoli et al., 2001 and 2003; Esonu et al., 2002, 2004 and 2005).

Considering the combinations of ingredients used by the traditional animal health practitioners, it is likely that additive, synergistic and nutritional effects of leaf meals might be involved in alleviating livestock maladies. Many phytochemicals have been shown to be bio-active, and have been useful as chemotherapeutic agents, pesticides, food/feed additives and other biologicals.

Congronema latifolia (utazi) is a wild tropical creeping plant with lush deep green vegetation. Where it grows in a swampy area or inland valley, the vegetation is perennial, otherwise, it is deciduous. The plant plays a significant role in ethnomedicine among the local populations where its leaf meal extract is used in treating malaria, stomach-ache and diarrhoea. Osuala et al (2005) reported that methanolic extract of C. latifolia exhibited marked activity against Pseudomonas aeruginosa and

E.coli with both having a minimum inhibitory concentration (MIC) of 15.625 mg/ml in an invitro study.  Their investigations have new evidence to corroborate an earlier work by Dalxiel (1956), which reported the use of C. latifolia in the treatment of diarrhoea.

Congronema latifolia is abundantly available in Cross River State, Nigeria. It can readily be found in its southern part, up to areas beyond the central region of the state. This location lies along longitudes 80 and 90  East, and latitudes 60 and 70 North of the equator, with a warm weather and an ambient temperature range of 21 – 300C.  It experiences an annual rainfall of 500 – 1070mm (Mfam, 2002). The forest areas of Akamkpa, Biase, Ugep, Mkpani, and Obubra among others are rich

reservoirs of the plant.

So far, studies on C. latifolia have been basically in vitro investigations. It is worthy of note that the nutritional and pharmacological principles of the plants oftentimes behave differently in the animal’s phisological environment from what obtains in in vitro studies. This is because the factor(s) under investigation may be inactivated in the in vivo environment, or such factor(s) may form complexes which limit or inhibit its bio-availability. Sometimes it may be flushed out of the system so rapidly that it has no time to act. At other times active ingredients in the plant materials can be poisonous or toxic at certain concentrations or administration regimen.

There is paucity of information on the nutritional, phytochemical and toxicological properties of C. latifolia. It would therefore seem necessary to undertake a comprehensive preliminary investigation of its leaf to determine its nutritional significance as feed ingredient in poultry and rabbit diets, as well as its phytochemical composition and potentials.

RESEARCH OBJECTIVES

The objectives of the research were therefore to:

Determine the proximate chemical composition of the leaf meal of

latifolia, including its mineral and amino acid profiles;

Test its bio-activity using common enteric bacterial organisms of poultry and rabbits.

Determine its phytochemical composition, and its

toxicological and histopathological effects using broiler chickens and crossbred rabbits;

Determine its effects on growth performance, organ weights, carcass weights, haematological and serum biochemical indices of the test animals and,

Provide reference data on its leaf meal as feed ingredient in poultry and rabbit diets.

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