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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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