Feeding Value of Fermented Pigeon Pea (Cajanus Cajan) Seeds with Carbohydrate Mixtures in Broiler Diets

Feeding Value of Fermented Pigeon Pea (Cajanus cajan) Seeds With Carbohydrate Mixtures in Broiler Diets

Najla Osman Mohammed Hamad

B.Sc. (Honours) Animal Production

University of Khartoum, 2000

A dissertation submitted to the University of Khartoum in partial

fulfillment for the requirement of the Degree of

Master of Science in Poultry Production.

Supervisor

Dr. Abbakar Ali Idriss

Department of Poultry Production

Faculty of Animal Production

University of Khartoum

September - 2006

DEDICATION

To my parents To my brothers and sisters To my friends I dedicated this work Najla

ACKNOWLEDGEMENTS

I wish to express my most sincere appreciation to Dr.

Abbaker Ali Idris. It would have been difficult to pursue this work with out his advice, his guidance, his encouragement and his patience and for aberrance.

Iam extremely grateful to the staff and members of Faculty of Animal Production, Department of Poultry Production, technical staff of the Biochemistry and Nutrition.

Finally special thanks are extended to my brother Dia,

Osama and Mohammed for their encouragement and patience.

ABSTRACT

This experiment was carried out of study the chemical composition and feeding value of fermenting combined to mixture of pigeon peas

(Cajanus cajan) seeds with (sorghum grain or sugar). Five mixing ratios of sorghum grains (20, 40, 60 and 80%) with one another mixing ratio (25%) of normal sugar were added to the substrates of pigeon peas seeds. The combined mixtures were soaked thoroughly in water and subjected to natural fermentation process until all of them were completely fermented.

The fermented products were sun dried and then some samples were taken for tannin analysis and proximate tannin determination chemical analytical.

On the basis of the chemical composition, the fermented products were used to formulate six dietary treatments where the total protein provided by the composition commercial super-concentrate of the broiler diets was replaced in each dietary treatment by protein of one of the fermented products.

A feeding trial through six week duration was conducted using 144 unsexed day old broiler chicks in an open–side deep litter poultry house.

Using complete randomized design, the formulated six dieting treatment were randomly assigned to the experimental birds in the prepared pens within the experimental house, so that each dietary treatment was replicate thrice where the experimental pen constitutes a replicate group of eight birds each.

The management processes throughout the experimental period include provision of optimum thermal and ventilation environment; ad lib watering and feeding; vaccination against Gumboro and New Castle diseases; in addition to performance records keeping.

The obtained results revealed that weight gain, feed intake, feed utilization efficiency, dressing % were significantly improved P< 0.05 with low levels of carbohydrates (20 – 25) sugar mixture. We concluded that low nutritional quality of the pigeon peas seeds (Cajanus cajan) can be enhanced by fermentation processes in the presents of low level of carbohydrates (20% sorghum grains or 25% sugar). ﻣﺴﺘﺨﻠﺺ اﻷﻃﺮوﺣﺔ ﺻﻤﻤﺖ هﺬﻩ اﻟﺘﺠﺮﺑﺔ ﻟﺪراﺳﺔ اﻟﺘﺮآﻴﺐ اﻟﻜﻴﻤﻴﺎﺋﻲ واﻟﻘﻴﻤﺔ اﻟﻐﺬاﺋﻴﺔ ﻟﺨﻠﻴﻂ اﻟﻠﻮﺑﻴﺎ اﻟﻌﺪﺳﻲ

واﻟﻜﺮﺑﻮهﻴﺪرات (ﺑﺬور اﻟﺬرة + اﻟﺴﻜﺮ) اﻟﻤﺨﻤﺮة.

ﺧﻤﺴﺔ ﻣﺴﺘﻮﻳﺎت ﻣﻦ اﻟﺬرة (0.0%، 20%، 40%، 60% و80%) ﻣﻊ ﻣﺴﺘﻮى واﺣﺪ

(25%) ﻣﻦ اﻟﺴﻜﺮ اﻟﻌﺎدي ﺗﻢ إﺿﺎﻓﺘﻬﺎ إﻟﻰ ﺑﺬور اﻟﻠﻮﺑﻴﺎ اﻟﻌﺪﺳﻲ. ﻏﻤﺮ اﻟﺨﻠﻴﻂ آ ﻠ ﻴ ﺎً ﺑﺎﻟﻤﺎء ﺣﺘﻰ ﺗﺨﻤﺮ

ﻃﺒﻴﻌﻴﺎً وﺟﻔﻒ اﻟﺨﻠﻴﻂ اﻟﻤﺨﻤﺮ ﺑﻮاﺳﻄﺔ أﺷﻌﺔ اﻟﺸﻤﺲ ﺛﻢ أﺧﺬت ﻣﻨﻪ ﻋﻴﻨﺎت ﻟﻠﺘﺤﻠﻴﻞ اﻟﻜﻴﻤﻴﺎﺋﻲ اﻟﺘﻘﺮﻳﺒﻲ

وﻗﻴﺎس ﻧﺴﺒﺔ اﻟﺘﺎﻧﻴﻦ.

إ ﻋ ﺘ ﻤ ﺎ د اً ﻋﻠﻰ اﻟﺘﺮآﻴﺐ ﻟﻠﻤﺎدة اﻟﻤﺘﺨﻤﺮة اﻟﻤﻨﺘﺠﺔ ﺗﻢ اﺳﺘﺨﺪاﻣﻬﺎ ﻟﺘﻜﻮﻳﻦ ﺧﻤﺴﺔ ﻋﻼﺋﻖ ﻣﺨﺘﻠﻔﺔ ،

ﺣﻴﺚ ﺗﻢ اﺳﺘﺒﺪال ﺑﺮوﺗﻴﻦ اﻟﻤﺮآﺰ ﻓﻲ آﻞ ﻋﻠﻴﻘﺔ ﺑﺒﺮوﺗﻴﻦ اﻟﻤﺎدة اﻟﻤﺘﺨﻤﺮة.

اﺳﺘﻐﺮﻗﺖ اﻟﺘﺠﺮﺑﺔ 6 أﺳﺎﺑﻴﻊ اﺳﺘﺨﺪم ﻓﻴﻬﺎ 144 آﺘﻜﻮت ﻻﺣﻢ ﻋﻤﺮ ﻳﻮم ﻏﻴﺮ ﻣﺠﻨﺲ داﺧﻞ

ﺣﻈﺎﺋﺮ ﻣﻔﺘﻮﺣﺔ ذات ﻓﺮﺷﺔ ﻋﻤﻴﻘﺔ – اﺳﺘﺨﺪام اﻟﺘﺼﻤﻴﻢ اﻟﻌﺸﻮاﺋﻲ اﻟﻜﺎﻣﻞ و إ ﻋ ﺘ ﻤ ﺎ د اً ﻋﻠﻰ اﻟﻤﻌﺎﻣﻼت

اﻟﻐﺬاﺋﻴﺔ ﻣﻊ وﺟﻮد ﻋﻠﻴﻘﺔ ﺿﺎﺑﻄﺔ ﺗﻢ ﺗﻮزﻳﻊ آﺘﺎآﻴﺖ اﻟﺘﺠﺮﺑﺔ ﻋﻠﻰ اﻷﻗﻔﺎص داﺧﻞ اﻟﺤﻈﻴﺮة ﺑﻮاﻗﻊ 8

آﺘﺎآﻴﺖ ﺑﻜﻞ ﻗﻔﺺ ﻣﻊ وﺟﻮد ﺛﻼﺛﺔ ﻣﻜﺮرات ﻟﻜﻞ ﻣﻌﺎﻣﻠﺔ.

ﻣﻦ ﺧﻼل اﻟﻤﻌﺎﻣﻼت اﻹدارﻳﺔ أﺛﻨﺎء ﻓﺘﺮة اﻟﺘﺠﺮﺑﺔ ﺗﻢ ﺗﺠﻬﻴﺰ اﻟﺤﻈﺎﺋﺮ ﺑﺪرﺟﺔ اﻟﺤﺮارة

اﻟﻤﻨﺎﺳﺒﺔ، اﻟﺘﻬﻮﻳﺔ اﻟﺠﻴﺪة ﻣﻊ ﺗﻮﻓﻴﺮ اﻟﻤﺎء واﻟﻐﺬاء ﺑﺼﻮرة داﺋﻤﺔ واﻟﺘﻄﻌﻴﻢ ﺿﺪ ﻣﺮض اﻟﺠﻤﺒﻮرو

واﻟﻨﻴﻮآﺎﺳﻴﻞ ﻣﻊ ﺗﺴﺠﻴﻞ اﻷداء اﻟﻴﻮﻣﻲ ﻟﻠﻜﺘﺎآﻴﺖ. أوﺿﺤﺖ اﻟﻨﺘﺎﺋﺞ أن هﻨﺎﻟﻚ زﻳﺎدة ﻣﻌﻨﻮﻳﺔ ﻓﻲ آﻞ ﻣﻦ

آﻤﻴﺔ اﻟﻐﺬاء اﻟﻤﺴﺘﻬﻠﻚ، اﻟﻮزن اﻟﻤﻜﺘﺴﺐ، ﻣﻌﺪل اﻟﺘﺤﻮل اﻟﻐﺬاﺋﻲ وﻧﺴﺒﺔ اﻟﺘﺼﺎﻓﻲ ﺑﺎﻟﻨﺴﺒﺔ ﻟﻤﺠﻤﻮﻋﺔ

اﻟﻜﺘﺎآﻴﺖ اﻟﺘﻲ ﺗﻐﺬت ﻋﻠﻰ ﻧﺴﺒﺔ ﻣﻨﺨﻔﻀﺔ ﻣﻦ اﻟﻜﺎرﺑﻮهﻴﺪرات (20% ذرة، 25% ﺳﻜﺮ) ﻣﻊ ﺑﺬور

اﻟﻠﻮﺑﻴﺎ اﻟﻌﺪﺳﻲ. ﺧﻠﺼﺖ اﻟﺪراﺳﺔ إﻟﻰ أن ﺗﺤﺴﻴﻦ اﻟﻘﻴﻤﺔ اﻟﻐﺬاﺋﻴﺔ اﻟﻤﺘﺪﻧﻴﺔ ﻟﺒﺬور اﻟﻠﻮﺑﻴﺎ اﻟﻌﺪﺳﻲ ﻳﺘﻢ ﻣﻦ ﺧﻼل

ﻋﻤﻠﻴﺔ اﻟﺘﺨﻤﻴﺮ ﻣﻊ وﺟﻮد ﻣﻮاد ﻣﺴﺎﻋﺪة ﻣﻦ اﻟﻜﺮﺑﻮهﻴﺪرات ﺑﻜﻤﻴﺎت ﺑﺴﻴﻄﺔ (20% ذرة أو %25

ﺳﻜﺮ). LIST OF CONTENTS

Page

DEDICATION………………………………………………………………………………… i

ACKNOWLEDGEMENTS……………………………………………………………… ii

ABSTRACT………………………………………………………….………………………… iii

ABSTRACT IN ARABIC ……………………..………………………………………… v

LIST OF CONTENTS…………………………………………………………………… vi

LIST OF TABLES………………………………………………………………………… viii

CHAPTER I: INTRODUCTION………………………………………………… 1

CHAPTER TWO: REVIEW OF LITERATURE ……………...………… 3

2.1 Nutritive value of Cajanus cajan…………………………………………… 4

2.2 Antinutritional factor of C. cajan………………………...………………… 8

2.2.1. Tannin……………………………………………………………………………..…… 11

2.2.2. Lectins…………………………………………………………………………….…… 12

2.2.3 Processing of C. cajan………………………………………………………...… 13

2.4 Rational for food fermentation…………………………………………..… 15

2.5 Advantage of fermenting legumes……………………………………….… 16

2.6 Variability of feed stuff………………………………………………………… 19 2.7 Use of C. cajan in poultry nutrition……………….………….…………… 19

CHAPTER THREE: MATERIALS AND METHODS…..……..……… 25

3.1 Fermentation process …………………….……………………...……………… 25

3.2 Chemical analysis…………………………….…………………...……………… 26

3.3 Experimental house and birds …………………………..…………………… 28

3.4 Experimental diets …………………………………………..…………………… 28

3.5 Management and data collection…………………………………………… 29

3.6 Experimental design and statistical analysis…………………………… 30

CHAPTER FOUR: RESULTS……………………………………………………… 34

4.1 Proximate analysis………………………………………………...……………… 34

4.2 Performance of broiler chicks……………...………………………………… 34

4.2.1 Feed intake………………………………………………………………...………… 34

4.2.2 Body weight gain………………………………………………………..………… 35

4.2.3 Feed conversion ratio (FCR) ………………………………………………… 35

4.2.4 Effect of dietary fermented C. cajan overall and 0% of broiler 36 chicks…………………………………………………………………...………………

CHAPTER FIVE: DISCUSSION………..………………………………………… 41

CONCLUSION AND RECOMMENDATIONS………………………….… 47

REFERENCES…………………………………………………….………………………… 48

LIST OF TABLES

Table Title Page

1 Proximate analysis of non fermented and fermented Cajanus cajan seeds……..………………………………………………….…………… 33

2 Composition of experimental diets as fed %……..………..…… 34

3 Calculated and determined chemical analysis of experim-

ental diets……..……………………………………..………………………… 35

4 Effect of dietary fermented C. cajan on average weekly

broiler feed intake (g/bird) ……..……………………………………… 39

5 Effect of dietary fermented C. cajan on average weekly

broiler weight gain (g/bird) ……..……………..……………………… 40

6 Effect of dietary fermented C. cajan on average weekly

broiler feed conversion ratio (FCR) (g/bird) ……..…………… 41

7 Effect of dietary fermented C. cajan on overall performa-

nce and dressing (%) of broiler chicks……..….………………… 42

CHAPTER ONE

INTRODUCTION

Food is the major cost in the layer and meat chicken industries. Over the past few years, world grain prices have fluctuated dramatically but overall have increased and continued to do so. Protein concentrates have also increased and at a more consistent rate.

The rapid expansion of the broiler industry and to a lesser extent the egg industry particularly in Asia and elsewhere has resulted in a rapid rise in demand for raw ingredients. There is likely to be a food shortage for livestock feeding in the next 10–15 years and during this time an intense competition for food resources and an escalation in food prices are predicted (Farrel, 1997; Van der Sluis, 1997). For the poultry industry alone, food requirement is expected to increase by 100 million metric tones

(Mt) between now and the years 2010.

Looking for alternatives, there have been a number of excellent reviews of the more commonly available legumes (Gatel, 1994; Wiseman and Cole, 1988); in addition to Petterson and Mackintoch (1994). In the

Sudan, Cajanus cajan which is a scientific name for a grain legume pigeon pea is currently considered as non-conventional feedstuff for poultry feeding. According to Salunkhe et al., (1985); Udedibie and Igwe, (1989);

Amaefule and Onwudike, (2000), the crude protein content of the raw pigeon seed ranged from 21 to 30%. A major advantage of grain legumes in the farming system is that they recycle and return nitrogen to the soil. Conversely, their major disadvantage is that most grain legumes contain one or more antinutritional factors (ANF) and this may limit their use as poultry feed or reduce the amount to be included in poultry diets (Huisman and Tolman, 1992) if it is untreated. Much research has been conducted to overcome the prevailing drawbacks and ANF of the grain legumes. So far, fermentation process is deemed as a common practice for improving the nutritional value of the grain legumes. Nevertheless, fermentation of grain legumes alone is found to be associated with development of very objectionable odors or health hazards. Fortunately, those reactions can be prevented or inhibited by addition of carbohydrates which act as co-factor for improvement of fermentation process (FAO, 2000).

This study is planned to determine the optimum combination ratio for fermentation of C. cajan and sorghum grains or sugar in an attempt to overcome the drawbacks of fermentation process of C. cajan alone and to improve the protein quality and functional properties of the fermented product. To evaluate the nutritive value, the fermented products, as protein source. CHAPTER TWO

LITERATURE REVIEW

Pigeone pea is a plant which is grown for human food. It’s wide range of cultivars and the many ways of their uses in farming system have made pigeone pea popular to small scale farmers. Pigeone pea is grown widely in

India and in some parts of Africa and Central America. The main producer of C. cajan. in the world include: India, Uganda, Tanzania, Kenya, Malawi,

Ethiopia, Mozambique, the Dominican Republic, Puertorica, the West

Indies in the Caribbean region and Latin American, Thailand, Indonesia, the

Philippines and Australia (Sinha, 1977).

Sudan has been identified as potential areas for C. cajan. The crop is grown in tropical and subtropical environments extending between 30°N and 25°S latitude with temperature range between 20°C and 40°C (Sinha,

1977).

Pigeon pea grows on acid sand in shale and alkali-clays in India. It is widely adoptable and more tolerant of high temperature and drought than most other crops. 2.1. Nutritive value of Cajanus cajan:

Protein is essential for growth and restoration of body tissues.itis

Particularly important in the diet of the mono - gastric animals. The protein content of commonly grown pigeone pea cultivars ranges between 17.9 to

24.30 /100 g (Salunkhe et al, 1986) for whole grain samples and between

21.1 and 28.1 g / 100g for split seed. In case of wild species of C. cajan, they have been found to be very promising source of high protein. For example,the report of Singh et al (1990) indicated that several high protein genotypes have been developed with a protein content as high as 32.5%.

These high protein genotypes were found to be 20% higher in their protein content than that of the normal genotype (Saxena et al., 1987 and Reddy et al; 1979).

The chemical composition of Cajanus cajan as shown by Sinha

(1977) contained about 57.3–58.7% carbohydrate, 1.2–8.2 crude fiber and

0.6–3.2% lipids. It is a good source of dietary minerals such as calcium, phosphorus, magnesium, iron, sulphur and potassium. Likewise, it is good source of soluble vitamins especially thiamine, riboflavin, niacin and choline. The Protein content of Cajanus- cajan ranges between 20.6–27%. As lysine was relatively high, methionine was relatively low with trypsin inhibitors 10.1mg/100gm and PER 0.9–1.40( Allies et al; 1978).

Gazzeta et al. (1995) worked on chemical composition and antitrypsin activity of hydrated and boiled green and mature seeds of

C.cajan. They found that nutrient content, ether extract and antitrypsin activity were higher in green seeds than mature ones. Magboul et al. (1976);

Nour and Magboul (1986) worked on approximate analysis of some legume seeds that grown in the Sudan. They found that C. cajan seed contain as percentage wise (6.1–6.2) moisture, (19.3–26.3) protein, (1.3–2.0) fat, (6.4–

8.2) fiber, (53.5–62.7) carbohydrates; in addition to (132–500 mg/g), calcium and (330–354 kcal/ kg), energy

Allies (1978) reported that C. cajan contained starch about 40.4%, moisture 8.71%, amylase 35.6%, Nitrogen 0.28%, fat 0.71%, and phosphorus 0.01%.

Chary and Bhlla (1980) studied the distribution of protein in different component of C. cajan and found that protein fractions of embryo include

17% albumin, 52.7% globulin, 21.3% glutein, 2.7% prolamin. Awadalla and AbdelAti (2001) worked on corticated and decorticated C. cajan on broilers nutrition and found the seed contained as percentage DM (91.3-

93.0), CP (31.5- 27.7), CF (11.8- 4.46), NFE (55.8- 58.7), Ash (5.0- 4.56),

E.E (2.18- 1.58), Tannin (0.32- 0.02) Ca (310- 280ppm), P (32- 29ppm), Fe

(50- 30 ppm) .

Regarding Amino acids and protein in C. cajan. Singh et al (1990) in accord to FAO/WHO (1984) provisional pattern stated that, methionine, cysteine and tryptophan constitute most important amino acids in C .cajan and therefore are expected to overcome their limitations in grain legumes.

They determined that the high protein genotypes contained significantly about 25% higher Sulphur- containing amino acids which are naturally methionine and cystein.

Singh and Eggum (1984) detected sulphur containing amino acids as percentage of C. cajan protein from 1.76 to 2.55%. The composition of some pulse grown in the Sudan including C. cajan Ahmed and Nour (1990) reported that sulphur amino acids had the lowest value for C. cajan. Copala

Krishna et al. (1977) worked in cajanus cajan amino acids and protein.

They mentioned that globulin fraction was considerably rich in sulphur amino acids. Elhardalou et al. (1980) detected the amount of amino acids in some legumes including C. cajan and found that tryptophan and methionine constitute the most deficient amino acids in C. cajan. Singh and

Eggum (1984), Pausztai (1986) and Singh et al (1984) worked on amino acids composition of C. cajan and determined that C. cajan to be rich in lysine (6.2–7.59 g/100g protein). Oshodi (1993) also studied amino acids content of C. cajan. He found that the seed contained essential amino acids but was low in sulphur containing amino acids. The work of Nowkelo

(1987) who studied the nutritional value of C. cajan meal and detected that amino acids content was low, especially cysteine and methionine, and that amino acids availability was 82.32% which is lower than amino acids availability of soybean meal (90%). C. cajan is more rich in none essential amino acid than sesame and Soybean (Awadalla, 2001).

With regard to fatty acids and mineral contents of C. cajan, Nwokolo

(1987) detected 69.4% fatty acids with low content of unsaturated fatty acid

(30.69%) and total absence of linolenic acid in addition to N-corrected metabolizable energy content of 11.08 mj/kg in raw C. cajan. He also worked in minerals and amino acids of C. cajan and reported a very high content of Calcium and Magnesium with low contents of Iron, Zinc, Copper and Manganese. His results also revealed average availability of minerals to be only 58%. Similar findings were reported by Sinha (1977) who mentioned that C. cajan is a good source of dietary minerals such as calcium (57-276), total P (131.8- 600ppm) and magnesium .

Regarding the vitamin contents, Sinha (1977) worked on Cajanus cajan vitamins and reported that seeds are the rich sources of soluble vitamins such as thiamin and choline. The chemical composition of C. cajan as reported Duke (1981) revealed carotene as vitamin A, 220 IU and vit B1, 150 IU per 100 g. The sun dried seeds of C. cajan were reported to contain (per 100g) B-carotene equivalent to 0.72 mg, thiamine, 0.14 mg, riboflavin, 2.9mg, the content of immature seeds of C. cajan on the other hand were reported to contain 145 IU B-carotene equivalent, 0.40 mg thiamine, 0.25 mg riboflavin, 2.4 mg niacin and 26 mg ascorbic acid/ 100g.

2.2. Antinutritional factor of C. cajan:

Legumes are important source of protein and energy for animals, however, inclusion of legumes for growing animals as the only source of protein almost leads to growth retardation and other undesirable physiological and biochemical alterations (Apata, 1989).

Aletor and Aldetimi (1989), Pausztai and Begbie (1989) found that plant proteins are more resistant to break down in alimentary tract than animal proteins because of the presence of antinutritional factors in the food of plant origin. They claimed that there are two main classes of protein antinutritional factors such as lectin and proteolytic enzyme inhibitors as trypsin inhibitors. Pauszati (1967) stated that there were some evidence that certain plant proteins were digested by pancreatic enzyme at slower rate than animal proteins. Several reviews discussed involvement of trypsin inhibitors and other toxic factors in the nutritional value of plant proteins

(Ologhobo and Apata, 1992: and Pezzato et al, 1995).

The role of trypsin and chemotrypsin inhibitors are reported to be antinutritional factors that are present in legumes (Singh and Jambunthan,

1980).

Lienar (1994) reported that C. cajan has lower levels of trypsin and chemotrypsin inhibitor activities as compared with Soybean. His report also reflected that there is a little information on the presence or absence of other undesirable components, in C. cajan such as oligosaccharide which are reported to cause flatulence.

C. cajan seeds as shown by Geri et al. (1994) contained at least nine trypsin inhibitors and four amylase isoinhibtors. They stated that these inhibitors have been extensively studied as antinutritional factors and as potential defense against pest. Trypsin inhibitor activity was greater in mature seeds, whereas green versus mature seeds differed little in their chemo-trypsin inhibitor activities (Lienar and Pichare, 1987).

The protein trypsin inhibitors, tannins, Phytate, phytic acid and phosphorous of C.cajan was studied by Ene-obong (1995). The author found the protein content of C. cajan to be 21–22.5% , trypsin inhibitors

7.5–14.4 mg/g and Phytate equal 8.13– 11.13 mg/g whereas phytic acid contributed 66.75% of phosphorous for C. cajan .He also added 77.8% as invitro protein digestibility coefficient for C. cajan.

Similarly, Sinha (1977) detected that the C. cajan seeds contained

12.7 mg/g phytic acid which constituted 78.2% of the total phosphorous and invitro protein digestibility coefficient of 60.4%. He denoted that there was significant negative correlation between phytic acid and invitro protein digestibility. Mahmoud et al (1986) studied the amylase and trypsin inhibitors in legume seeds. They concluded that, the amyloletic inhibitors have protein characters whereas the trypsin inhibitors are water solubles.

Mitijaval et al (1987) reported that. C. cajan contained tannin range of 0.0–0.25. There is evidence that the seeds of C. cajan contained considerable amount of Polly phenolic compounds, which may or may not be tannin. The Polly phenols are found to inhibit to large extent the activities of trypsin, chemotrypsin and amylase. The Polly phenolic compounds in C. cajan are adversely affected the activities of digestive enzymes and this affect on nutrient utilization. Seed coat contained the highest proportion of tannin, whereas red seeds have higher levels than the white ones, and this may affect some of digestive enzymes (FAO, 1976).

The possession of C. cajan to polyphenolic compounds such as tannine was also reported by Igbedion et al. (1995). They claimed that the black seed of C. cajan contained the highest proportion of tannins and that the red seeds had higher levels of tannins then the white seeds.

The growth depressing effect of tannin and toxicity in chicks has been studied by Kratzer et al (1968). They illustrated that growth of chicks reduced when they were fed a ration containing as little as 0.5% tannic acid.

Moreover, a level of 5% caused chicks mortality of about 70% between 7–

14 day old, and that tannin acid reduced the metabolizable energy (ME) of the diet and nitrogen retention in chicks.

Lectins constitute specific class of proteins widely distributed in nature. Diets based on raw legume seeds usually contain lectins some of which may possess strong anti-nutritive properties. Although some lectins can be inactivated by proper heat treatments (30–40°C). One of the main reasons why lectins possess strong antinutritive properties is due to its extra ordinary degree of their resistance to proteolytic breakdown in the gut

(Pauztai and Begbie 1989).

Shama et al (1995) worked on C. cajan lectin. They isolated lectin from seeds by ammonia soleplate fraction. The lectin was admirely composed of identical subunit with N–and C-terminal residues of theroinine and alanine. The glucoprotein lectin contained 3% concavaline, plus are active natural carbohydrates. C. cajan lectin was specific manose and glucose.

2.3. Processing of C. cajan:

Mulimani et al (1994) stated that, soaking for 24 hours followed by cooking for 20 minutes was more effective in destroying the trypsin and chemo-trypsin inhibitory effects.

Igbedioh et al. (1995) monitored the effect of processing and autoclaving on total phenols and chemical composition of C. cajan. They stated that, cooking of soaked and dehulled seeds lowered total phenols contents of C. cajan by 49%. Whereas sprouting of legumes seeds increase protein content and reduced the carbohydrate content. Rani et al (1996) studied effect of boiling and soaking of C. cajan seed and they concluded that, soaking to 12hr at 37°C and/or boiling reduced trypsin inhibitor activity and improved protein (pepsin) and starch digestibilities.

Chitra et al (1996) studied the effect of various type of processing on

C. cajan. They found that germination decreased phytic acid content by

50% whereas autoclaving and roasting are more effective in decreasing phytic acid. Fermentation on the other hand was found to cause great increase in vitro protein digestibility.

Awadallah (2000) detected that decortications of legume seeds increased, NFE, sulphur and DM; however, seed CP, CF and EE were decreased as well as tannin content was also decreased to 0.02% comparing to 0.32% in corticated seeds. Ahmed and Mohamed (2003) noted that heat treatment reduced trypsin inhibitors, lectin, HCN and tannin content of guar seeds. Mulimani and Daramiyothi (1995) observed that trypsin and chemotrypsin inhibitor activities were reduced by soaking on salt solution in comparison to soaking in distilled water. Both of the latter two processing methods altered the composition of legumes without causing any change in carbohydrate. McnNab and Longstaff (1991) reported that, digestibility of amino acids; starch and lipid of a diet with tannin free hulls of faba bean were lower than those of a control diet. They claimed that dehulling will not reduce all antinutritional factors, because of the fact that lectin and trypsin inhibitors are concentrated in the cotyledons.

2.4. Rationale for food fermentation:

According to Steinkraus (1996), fermentation plays at least five important roles in the diet of the developing fowls. He countered that:

1. Dietary enrichment through development of diversity of

flavours, aromas and textures in food substrates.

2. Preservation through lactic acid, alcoholic, acetic acid

and alkaline fermentation.

3. Biological enrichment of food substrate with protein,

essential amino acid, essential fatty acids, and vitamins.

4. Detoxification.

5. decreasing cooking time and fuel requirement.

Microorganisms associated with indigenous fermented foods are generally elided. Those with ability to produce enzymes, vitamins, essential amino acids, essential fatty acids, antibiotics organic acids, peptides, protein, fat, complex-polysaccharides, desirable flavor compound, and\or flavor–enhancing compounds, are of potential value to the food industry.

The cultures and strains associated with food fermentation offer potential for genetic modification and genetic engineering in the future.

2.5. Advantage of fermenting legumes:

Fermentation offers several advantages over other food processing and preservation procedures. Fermentation utilizes microorganisms for the transformation of row materials into useful products. It often results in the production of nutritionally enriched very stable food products from low value carbohydrate and protein substrates. Fermentation and particularly in the developing countries is generally less expensive than other forms of food preservation.

In tropical regions environmental conditions often lead to rapid food spoilage. Fortunately, fermentation was found to improve or salvage nutrition in such food stuffs. Fermentation often results in new improved flavors, and textures in the food product. Fermentation also leads to improvement in nutritional value of feed through enrichment with microbial protein, amino acids, lipids and vitamins which are sometimes not present in the original substrate. Unlike western fermented food, many fermented oriental products utilize cereal–legume combination. Food fermentation has natural and/or mixed culture fermentation and incorporates a number of different species and genera of yeasts. Fungi and/or bacteria, microbial interaction varies in accordance with the type of fermentation and are quite complex for many indigenous fermentation. Aidoo, (1986) reported that fermentation process modified the physical, nutritional and sensory status of the raw material to yields products of overall improved quality in grain legumes. Odunfa,

(1983a; 1985a) detected that raw legumes and nuts contain substances such as tannin, trypsin and chemotrypsin inhibitors which reduce protein digestibility,these antinutritional factors are reduced to save levels through microbial activity in fermentation process. The fermentation of grain legume,seeds and nuts involve the activation of some enzymes, particularly lipolytic and proteolytic which enhance the digestibility of raw plant material (Achinewhu, 1983b).

Moulds are some of the most important organisms associated with the fermentation of grain legumes, seeds and nuts in the Asia-Pacific region. Koji which is starter used in the production of a variety of fermented products, is produced by mould fermentation (FAO, 2000).In fermentation of cereal/grain-legume combination, a number of studies have shown that there was a complementary protein effect between cereal grains and legumes (Bressani, 1993). He also detected that this effect has been attributed to the contribution of lysine to the cereal grain by grain legume protein and to the contribution of the methionine from cereal proteins to grain legume protein. The proximate composition of most foods does not change significantly during fermentation, however, the soluble fraction of food is increased, microbial proteases degraded complex protein into simple proteins, peptides and amino acids, whereas the break down of complex carbohydrates also result in an increase in soluble simple sugars as reported by Achinewhu (1983a). Elkhalifa (1993) reported that fermentation of the high-tannin sorghum grain sorghum relatively depleted most of its nutritionally important fractions could explain the tendency of fermentation to reduce the protein contents of the sample.

The distribution and physiological effects of antinutritional factor (ANF) in grain legumes:

ANF Distribution Physiological effect

Protease inhibitors Most legumes Depressed growth pancreatic, hyper plasta

acinar nodules interference with protein

digestion.

Tannin Most legumes Interference with protein and starch digestion.

Amylase inhibitors Most legumes Interference with starch digestion.

Lectins Most legumes Depressed growth , death

Oligosaccharids Most legumes Flatulence

Saponius Most legumes Affect intestinal permeability

Phytate Most legumes Interference with mineral availability.

Non starch Most legumes Depressed digestibility of protein, starch and fat. polysaccharide (NSP)

Source: Wiryawan and Dingle (1999) 2.6. Variability of feed stuff:

The processed C. cajan seeds are currently considered as non- conventional feed stuff in poultry feeding and as available protein feed resource (Preston, 1987; Udedibie and Igwe, 1989; Amaefule and Obioha,

1998; Amaefule and Onwudik, 2000). World feed resources are diverse and their nutritive value may vary tremendously according to the source and other factors. It is well known that the chemical composition of feed ingredients is affected by varieties. For example, Ishag (1986) indicated that while variety affected DM, CP and CF content, location influenced EE, CP and NFE of sorghum. Similaly, sampling affected dry matter and milling affected both EE and CP of wheat bran. Like wise Miller et al., (1994) reported the grain legumes in terms of proteins, essential amino acids, minerals and ash content are differed according to the varieties and location of plant. Also, climate and soil variety are found to be the main determinant of yield and percentage of protein in the crop, (Shellcanberger et al., 1995).

2.7. Use of C. cajan in poultry nutrition:

The first use of C. cajan in poultry feed was conducted in Hawaii by

Drapper (1944). He replaced 45% of commercial feed which contain

(barely, cracked corn, alfalfa and soy bean) by C. cajan seeds for laying hens). The result indicated that relatively high rate of C. cajan can be included in layers ration ,egg production and mortality was not affected by inclusion of C. cajan in layer rations. Agwanobi (2000) detected the feeding value of heat treated soybean and C. cajan rations in comparison with standard ration. The performance of layers fed on the three experimental diets was monitored for 56 days. Egg production, egg weight, Feed intake and feed utilization efficiency were lowest for C. cajan ration; however, soybean gave the same level of performance as the standard ration. There were no significant difference between the rations with respect to albumen quality, and shell thickness. It is concluded that, C. cajan unlike soybean can not satisfy the protein requirements for optimum layer performance.

Pezzate et al. (1997) studied the substitution of soybean by C. cajan in broiler chicks. They fed day old broilers diets containing: 10, 15, 30, 43%

C. cajan meal at the expense of soybean meal. They found that substitution of 15% soybean meal by C. cajan is more effective for broiler in terms of feed conversion ratio, live weight and feed intake.

Elias et al. (1996) studied the effect of cooking in nutritive value of and antinutritional factors of Phaseolus vulgaris, cowpea, C. cajan and soybean. They found protein digestibility of cooking was low in all species where as lysine was relative high and methionine was relatively low.

Trypsin inhibitors were also found to be variable among species.

Mizubuti et al. (1995) fed broilers dietary treatments based on maize and soybean meal. A basal diet containing 20.1% CP and ME 2910kcal/kg with 0, 10, 20, 30, 40 or 50% raw C. cajan seeds from 1 to 28 days old and from 29 day to 45 days. The obtained results showed that there were no difference in gain among treatments, feed conversion was best for the control diet.

Leon et al. (1993) worked on force feeding of the legumes and used

Vigna ungniculata, Posphocarpustition genolobus and C. cajan seeds containing 26, 37 and 20% CP; trypsine inhibitors activities were 12.5, 68 and 18.7mg/gm, protein of 50%, legume were force fed to 48 Rod Island red cockerels, which were deprived of feed for 48h before and after force feeding. The collected data revealed in respective variation, nitrogen digestibility values among the legumes (79, 54 and 81%) and in true ME

(2913, 2512 and 2832 kcal/kg DM).

Tangtaweewipat and Elliot (1991) conducted two experiments to evaluate the effect of feeding broilers and layers raw C. cajan seeds. In the first experiment they fed C. cajan to broilers through seven weeks to come up with the following observations: dietary treatments containing C. cajan from zero%to500g/kg showed no significant variation in their support to live weight gain .Moreover, the lower the dietary level of C. cajan (o.o-

300g/kg) the better FCR were obtaind. In the second experiment, they included C. cajan into layers diets (16% CP ME 11.9 Mj/kg), to replace maize and soybean meal. The effect of C. cajan on egg production and egg quality over 3 monthes showed that C. cajan at 200g/kg or greater decreased egg production and feed conversion ratio but did not affect feed intake or egg quality.

To study the substitution of C. cajan seeds for groundnut cake and maize in broilers finishing diets, Amaefule and Obioha (1998) fed broilers diets containing 0, 30, 40 or 50% C. cajan seeds meal. Feed intake and protein intake increased (P < 0.05) as a result of replacing groundnut cake and maize with CCSM up to 40% but did not show significant difference between 40 and 50%. Feed intake figures were 96.80, 129.90, 144.30 and

142.60 gm and the corresponding protein intake figures were 20.67, 27.64,

30.72 and 30.57 for 0, 30, 40 and 50% inclusion rats of CCSM, respectively. FCR and PER did not differ among the experimental groups. Ologhobo, (1992) conducted two experiments to study the nutritive values of some tropical legumes for poultry nutrition. In the first study, seven days old broiler chicks were fed on maize based diets in which 12.5 or 25% of the soybean meal was replaced by jack beans (Canavalia ensiformis), Kidney beans (Phaseolus vulgaris), lima beans (Phaseolus lanatus), yam beans (Pachyrhizus erosus), Pigeon peas (Cajanus cajan) or bambarra groundnuts (Vigna subterranea). Inclusion of legumes at 12.5% resulted in significant decrease in weight gain and feed intake. In the contrary, feed intake (kg) per kg weight gain for broilers given diets containing 12.5% lima beans or pigeon peas, and 12.5 or 25% bambarra groundnuts or yam beans did not differ significantly from the controls.

Similar levels of jack, kidney and lima beans or pigeon peas at 25% adversely affected feed utilization efficiency whereas phosphatase enzyme activities were increased at the higher levels of legume. It is concluded that only pigeon peas and bambarra groundnut can be recommended for inclusion in poultry diets and care should be taken if they are to be fed over long period. Likewise, toasted for 30 minutes, boiled for 30minutes or soaked in water for 24 hours. The results showed that pullet’s fed 10% raw or processed pigeon peas seeds meal (PSM) diets did not differ significantly in all performance parameters. It was concluded that PSM is a good source for pullets and that 10% raw or processed PSM could be in corporate into pullet chick diets.

CHAPTER THREE

MATERIAL AND METHODS

The undertaken experiment was carried out in the premises of

Department of Poultry Production, Faculty of Animal Production of

Khartoum University. The experiment was commenced in October and continued to November 2005. The ambient temperature during the experimental period ranged between 23° to 38°C.

3.1. Fermentation process:

The seed of Cajanus cajan used in the study were purchased from local market of Omdurman. At home, the bought seeds were cleaned, washed and then covered with water in a plastic container. After 12 hours soaking period, the soaked seed were washed under tab water and then divided into 5 prepared plastic containers to be mixed with either 25% sugar or different levels (20, 40, 60 and 80%) sorghum grains.

The combined mixtures of the C. cajan seeds with carbohydrate within each bottle were thoroughly mixed and soaked in water for a comple of days until all of them were completely fermented. Post completion of the fermentation process, each mixture was spread on a plastic sack and kept under sunlight for 2 – 3 days until all of them got dry. 3.2. Chemical analysis:

Samples were drown from each fermented mixture and non fermented cajanus seeds for proximate analysis and tannin determination.

The proximate analysis were analyzed according to AOAC (1980) procedure. The metabolizable energy values were calculated according to the modified equation of Lodhi et al. (1976) where they stated that ME =

(1.549 + 0.102 CP + 0.275 EE + 0.148 NFE – 0.0034 CF X 239).

The tannin contents of the samples were determined by the modified vanillin HCl methods of Price et al. (1978). The method requires transference of 200 milligrams of a sample into a conical flask. This step is followed by addition of 10 ml of methanol. The flask should be shaked in an orbital shakers for 20 min. and followed by filtration. One ml of the supernatant to be piptted into a test tube and 5 ml of vanillin HCl reagent will be added. The reagent is prepared by mixing equal volumes of 8% concentrated HCL in methanol and 2% vanillin in methanol. The two solutions of regents were mixed just prior to their use. After addition of the reagent, the sample was left for 20 minutes allowing the colour to develop and optical densities were obtained by using Bachman model colorimeter at 500 nm. A stock solution of (+) catchin was prepared by addition of 100 mgs of (+) catchin to 50 mls methanol.

From the stock solution, a series of dilutions were made to draw the standard curve. 1mL of each of the diluted series was piptted in the separate test tube and treated according to the methods described above.

When determining the tannin content of the fermented C. cajan, l ml of the extract was added to 5 ml of methods and this was used as a control to correct the optical density values of the fermented grain samples from colours due to other pigment. The tannin content is calculated as catchin

Equivalent (CE) as follows:

CE = C ×10 × 100 % 200 mg

Where:

CE: catchin equivalent

C: concentration corresponding to the optical density

10 ml: volumes of the extract

200 mg: weight of sample in mg

The chemical composition of the analyzed samples were shown in

Table 1. The fermented legume-carbohydrate mixtures were used at the expense of the commercial super concentrate of the standard broiler diet to formulate five experimental diets. 3.3. Experimental house and birds:

The experiment was carried out in open-side deep litter poultry house. The house was partitioned into 18 pens (1×1m) with enough working space allowances. The house was cleaned, washed and disinfected using formalin. Each pen was provided with feeder and drinker light was supplied 24 hours a day by natural light during the day and artificial light during the night.

A total of 144 unsexed day old broiler chicks (Ross) were obtained from Ommat company from Egypt. The chicks were weighed and then allotted randomly and evenly into the prepared 18 pens in-groups of 8 chicks per pen. The experimental pens were then randomly assigned to the six dietary treatments. The dietary treatments were replicated three times and each experimental pen constitutes a replicate group.

The birds were vaccinated against Gumboro and New castle disease at 2 and 3 weeks of age respectively.

3.4. Experimental diets:

Six experimental diets were fermented according to the chemical composition of the fermented product with graded level of fermented C. cajan to meat the requirements of broiler chicks outlined by the National Research Council, (NRC 1984). The description of formulated dietary treatments is shown in Table (2) whereas Table (3) showed the calculated and determined chemical composition of the experimental diets.

3.5 Management and data collection:

The experimental birds were kept accessible to water and feed throughout the experimental period. Drinkers were cleaned and refilled twice a day; feeds were weighed and added to the feeders wherever it is necessary. In order to minimize the feed loss, feeds were kept within the feeders at depth no more than the 2/3 of the feeder. Feed intake and body weight gain were determined on weekly basis. The recorded data were used for estimation of feed conversion ratio (FCR). Mortality rates and clinical signs were registered wherever they occurred. At the day of termination, birds were fasted for 12 hours in order to empty their digestive tracts and to reduce the chance of contamination during cleaning of ingested matter from digestive tract.

Chicks from each dietary treatment were leg banded, weighed individually and slaughtered. The birds were scaled in a pot of boiling water and feathers were plucked manually. The carcass were washed and allowed to drain. They were eviscerated by ventral cut and finally they were weighed. The weighed carcasses were used for determination of the dressing out percentages.

3.6 Experimental design and statistical analysis:

A complete randomized design was used in the experiment, the data generated from the experiment were subjected to analysis of variance, such reference cited in other should be stated clearly.

Table (1)

Proximate analysis of non fermented and fermented Cajanus cajan seeds

Component Experimental diets

% NF F1 F2 F3 F4 F5

D.M 93.80 96.20 95.20 96.20 96.15 96.40

C.P 26.70 23.50 20.00 18.90 16.00 23.10

E.E 1.80 2.90 1.15 2.00 1.50 2.20

N. F. E 58.50 58.40 70.15 62.30 70.12 61.40

C. F. 8.80 7.90 3.00 4.40 6.30 7.10

Ash 5.00 3.40 2.05 2.20 2.00 4.50

Tannin 0.52 0.00 0.40 0.40 0.50 0.00

M.E. 3137.16 3135.22 3390.32 3130.32 3287.93 3192.00 Kcal/kg(2)

NF: Non fermented C. cajan.

(1): F1: 80 : 20% legume : sorghum ratio, F2: 60 : 40% legume : sorghum ratio,

F3: 40 : 60% legume : sorghum ratio, F4: 20 : 80% legume : sorghum ratio, F5 75 : 25% legume : sorghum ratio. (2): ME is determined according to equation of Lodhi (1970).

Table (2)

Composition of experimental diets as fed %

Experimental diets(1) Component % Control F1 F2 F3 F4 F5 Sorghum grains 64.00 57.64 55.88 54.61 51.86 55.77 Groundnut meal 22.50 20.50 21.057 21.57 21.57 21.14 Sesame meal 6.80 9.30 9.00 9.00 10.00 10.00 Fermented C. cajan 0.00 8.50 10.00 10.50 12.50 8.50 Super concentrate(2) 5.00 - - - - - d. cal 1.40 1.76 1.76 1.76 1.76 1.76 O’shell 0.00 0.80 0.80 0.80 0.80 0.80 NaCl 0.25 0.25 0.25 0.25 0.25 0.25 Lysine 0.00 0.50 0.50 0.50 0.50 0.50 Methionine 0.02 0.15 0.15 0.15 0.15 0.15 Veg. oil 0.00 0.60 0.60 0.50 0.70 0.70

(1): F1: 80 : 20% legume : sorghum ratio, F2: 60 : 40% legume : sorghum ratio,

F3: 40 : 60% legume : sorghum ratio, F4: 20 : 80% legume : sorghum ratio, F5 75 :

25% legume : sorghum ratio, F5 75 : 25% legume : sorghum ratio. (2): ME= 2100kcal/kg, CP= 40, CF= 2, CFib= 2, Ca= 10, P= 4, Lysine= 12, Methionine= 3, Methionine + Cystine = 3.2

Table (3): Calculated and determined chemical analysis of experimental diets Experimental diets(1) Ingredients % Control F1 F2 F3 F4 F5 Calculated C.P. 23.10 23.00 23.07 23.00 23.00 23.40 C.F. 4.40 4.70 4.50 4.60 4.90 4.80 M.E. kcal/kg2 3105.60 3120.40 3127.90 3112.30 3120.60 3115.10 Lysine 1.08 1.08 1.11 1.12 1.14 1.10 Methionine 0.45 0.47 0.48 0.48 0.49 0.48 Methionine + cys 0.71 0.74 0.75 0.75 0.77 0.75 Determined D.M 94.58 94.24 94.20 94.33 94.42 94.61 C.P 26.60 25.90 23.10 22.02 21.35 24.85 C.F 6.30 6.30 6.40 7.10 6.40 5.80 E.E. 3.40 3.50 3.40 3.90 3.00 2.60 Ash 8.00 7.60 7.10 6.50 6.70 8.10

(1): F1: 80 : 20% legume : sorghum ratio, F2: 60 : 40% legume : sorghum ratio,

F3: 40 : 60% legume : sorghum ratio, F4: 20 : 80% legume : sorghum ratio, F5 75 : 25% legume : sorghum ratio. (2): ME is estimated in according to equation to Lodhi (1976). CHAPTER FOUR

RESULTS

4.1 Proximate analysis:

The results of proximate analysis of non-fermented (NF) and fermented (F) C. cajan seeds were shown in Table (1). It reveals that fermentation process increased dry matter and nitrogen free extract. On the other hand, crude protein, crude fiber, ash and tannin contents were decreased.

4.2 Performance of broiler chicks:

4.2.1 Feed intake:

The effect of feeding different ratios of fermented C. cajan- sorghum mixture (FCSM) on weekly broiler feed intake was shown in Table (4).

Dietary treatments had no significant (P> 0.05) effects on feed intake during the first three weeks, however, the results revealed significant (P<

0.05) variation in feed intake during 4th – 6th week of age. Chicks fed the control diet or diet containing the fermented product consumed more feed than their counterparts that were subjected to diet containing fermented products with higher levels of sorghum grains (40-80%). It's obvious that feed intake increased with the high level of C. cajan in fermented diet.

4.2.2 Body weight gain:

Result of weekly body weight gain were given in Table (5). Dietary treatment had no significant effect (P> 0.05) during the first three weeks.

Through the last three weeks, there was significant (P< 0.05) effects. Birds fed the control diet or diet with low carbohydrates levels of fermented products generally showed significant (P< 0.05) accelerated growth rate than those imposed to diet containing high carbohydrates levels of the fermented mixture products.

4.2.3 Feed conversion ratio (FCR):

Table (6) showed weekly feed conversion ratios. Dietary treatments had no significant (P> 0.05) effect on FCR during the first four weeks of age. However, the FCR was significantly (P< 0.05) affected by the dietary treatments during the finisher phase. The control and dietary treatments with lower levels of carbohydrate mixture for fermentation (20-40%) revealed better feed utilization efficiency than the dietary treatment with higher carbohydrates mixture (60-80%). 4.2.4 Effect of dietary treatments on overall performance of broiler chicks:

The influence of dietary treatments on overall performance and dressing percent are summarized in Table (7). The results revealed that there was significant (P< 0.05) effect between treatments. The chicks fed the control diet as well as the dietary treatments that contained low carbohydrates levels of the fermented mixture products were in general significantly (P< 0.05) excelled over their counterparts in terms of feed intake, body weight gain and feed utilization efficiency.

With respect to dressing out percentages, although the obtained results were not significantly (P> 0.05) affected by the dietary treatments, the highest dressing out percentages were correspond ant with the control or dietary treatments containing low carbohydrates levels of the fermented products.

Table (4).

Effect of dietary treatments on average weekly broiler feed intake (g/bird).

Dietary treatments

F1 F2 F3 F4 F5 Age NF C : S C : S C : S C : S C : Sg ±S.E. control 80-20% 60-40% 40-60% 40-80% 75% : 25% 1 110.41 100.00 106.25 110.41 112.50 102.08 6.07

2 278.12 264.58 258.58 270.83 240.16 262.03 13.97

3 422.87 482.00 480.41 480.87 492.83 496.25 14.25

4 625.99a 644.66a 500.83b 480.54bc 453.17bc 621.25a 24.06

5 652.49a 616.66a 582.91b 514.16b 545.71b 606.87a 24.36

6 945.83a 819.53ab 722.43bc 561.90c 593.64c 821.94ab 59.77

Total 3035.71a 2927.40a 2651.40b 2418.00d 2438.00d 2910.42a 90.37

1- C : S Mean C. cajan : sorghum ratio. 2- C : S Mean C. cajan : sugar ratio. 3- Values are means of 3 replicates, 8 birds/each 4- Means on the same raw sharing common superscripts not significantly different at 0.05% level. 5- S.E.: Standard error of means.

Table (5):

Effect of dietary treatments on average weekly broiler weight gain (g/bird).

Dietary treatments

Age F1 F2 F3 F4 F5 NF (weeks) C : S C : S C : S C : S C : Sg ±S.E. control 80-20% 60-40% 40-60% 40-80% 75%: 25% Initial Bwt 42.30 44.60 42.50 41.90 45.20 42.40 1 60.08 56.29 55.12 53.83 55.38 58.71 2.60 2 156.80 141.40 139.70 131.20 137.50 149.70 12.90 3 244.70 245.80 247.80 223.50 224.90 263.60 20.10 4 349.00b 390.40a 335.70ab 309.70bc 269.60c 341.95ab 29.20 5 528.70a 503ab 390.20bc 258.30c 236.00c 435.90ab 34.60 6 503.60ab 649.20a 449.50b 340.70c 321.70c 625.00a 47.00 Total 1845.00a 1895.39a 1618.02b 1317.50c 1260.61c 1859.43a 74.14

Table (6).

Effect of dietary treatments on average weekly broiler feed conversion ratio

(FCR) (g feed/ g Bwt).

Dietary treatments

F1 F2 F3 F4 F5 Age NF C : S C : S C : S C : S C : Sg ±S.E. control 80-20% 60-40% 40-60% 40-80% 75% : 25% 1 1.80 1.70 1.90 2.00 1.90 1.80 0.13 2 1.80 1.80 1.80 2.00 1.60 1.90 0.16 3 1.70 1.90 1.90 2.10 2.10 2.30 0.16 4 1.70 1.60 1.40 1.50 1.60 1.80 0.27 5 1.20c 1.20c 1.40bc 2.30ab 2.70a 1.40bc 0.25 6 1.80a 1.40 c 1.60ab 1.60ab 1.80a 1.30c 0.45 Total 1.60bc 1.50c 1.60bc 1.80ab 1.90ab 1.60bc 0.10

Table (7).

Effect of dietary treatments on overall performance of broiler chicks

Dietary treatments

F1 F2 F3 F4 F5 Parameters NF C : S C : S C : S C : S C : Sg ±S.E. control 80-20% 60-40% 40-60% 40-80% 75%: 25%

Feed intake (g/bird) 3035.00a 2927.40a 2651.40b 2418.00d 2438.00d 2910.42a 90.37

Weight gain (g/bird) 1845.00a 1895.39a 1618.02b 1317.50c 1260.61c 1859.43a 74.14

Feed conversion 1.60bc 1.50 c 1.60bc 1.80 ab 1.90 a 1.60bc 0.095 ratio

Live body weight 1803.33a 1850.00a 1400.00bc 1203.33c 1416.66bc 1817.00a 31.79

Dressed out (g) 1416.66a 1466.66a 866.66b 816.66b 916.66b 1300.00a 44.09

Dressing (%) 78.55 79.27 61.90 67.86 64.7 77.04 2.64

Mortality (%) 0.00 0.00 0.00 0.00 0.00 0.00

CHAPTER FIVE

DISCUSSION

On the basis of the proximate chemical analysis of the non fermented

Pigeon peas seeds versus the fermented mixtures of pigeon peas seeds with carbohydrates (sorghum grain and sugar), the substrate of the non fermented pigeon peas seeds recorded 26% crude protein which is within the range that have been widely reported by other researchers (Borget,

1992; Amaef and obioha, 1998). In case of the combined pigeon peas with carbohydrates , the fermented products generally depicted a drop in their crude protein, crude fiber ,moisture and mineral contents but there was an increase in their dry matter and nitrogen free extracts . The observed decline in chemical constitutes of the fermented pigeon peas- carbohydrates mixtures is possibly due to the dilution effects of the incorporated carbohydrates since the decreasing or increasing magnitudes of the breading chemical elements are almost proportion with the carbohydrates inclusion rates of the fermented mixture products. The obtained results also revealed that with the exception of the increased nitrogen free extracts of the fermented products, the rest of the result tend to differ from the findings of paredes–lopez and Harry (1989) who reported that the fermentation process decreased fat and fiber contents but increased soluble solids, total and soluble proteins, soluble carbohydrates and the PH of the grain legumes.

This variation could be attributed to differences in fermentation period, temperature, PH and even the variety of the grain legume used or type of treatment imposed to.

Regarding the anti nutritional factor, it is interesting to be noted that addition of carbohydrates at low levels (20-25%) to pigeon peas seeds pre the fermentation process provided tannin free fermented products (0.00vs

0.40- 0.50%). This result could point out one of the improving action of fermentation process to nutritional quality of grain legumes when combined with carbohydrates. The role of fermentation process in reducing anti nutritional factors in feedstuffs was documented by Achinewua in 1983b.

The decreased tannin contents of the fermented products agreed with the justification of the Paredes-lopez and Harry (1989) who attributed the reduction in tannin level of the fermented products to the treatments that have been applied during substrate preparation such as socking that precedes the fermentation process.

With respect to chick performance or biological responses of the experimental bird which include feed consumption rates, body weight gain, feed utilization efficiency, livability and dressing out percentages, it is obvious that there is no significant variation in birds response to any of the dietary treatments through out the starting phase. The lack of significant variation in bird response to the experimental diet during the first three weeks of life suggests that the dietary treatments to be similar in their physical and chemical properties. Aidoo (1986) pointed out that fermentation process can modify the physical, nutritional and sensory status of the raw material to yield of improved quality in grain legume. The performance responses of the bird fed dietary treatments containing the processed pigeon peas with the birds under the control diet during the starting phase of broiler chicks consolidates the results of D’mello (1995) who attributed the drop in f eed intake and body weight gain of birds fed on diet containing unprocessed pigeon peas to possession of raw pigeon peas some antinutritional factors. In other words, the obtained results indicates that fermentation as a processing mean will eliminate the fermented products from antinutritional factors of the original feedstuff. Likewise, the lack of mortality among the experimental birds proposes the freedom of the diets containing the fermented products from harmful substances or pathogenic organisms. This latter emphasis is in line with the work of Zamora and Fields (1979a) who claimed that the natural fermentation process is lactic in nature and therefore the fermented products are usually free from pathogenic contaminates. Similarly, Steinkraus (1996) defined the fermented foods as food substrates that are invaded or overgrown by edible microorganisms whose enzymes, particularly amylases, proteases, lipases hydrolyze the polysaccharides, proteins and lipids to nontoxic products with aromas flavor and textures that make them more pleasant and attractive to the human consumer.

In contrast to the starting phase, the responses of the experimental birds to dietary treatments during the finishing period showed significant discrepancies (P < 0.05). For instance, birds subjected to dietary treatments containing low levels of carbohydrates (20–25 vs. 40–80%) showed (feed intake,body weight gain and feed convesion ratio) similar to that of the control group which is imposed to the standard commercial broiler diet. The ability of the dietary treatments containing fermented pigeon peas with low levels of carbohydrates (20–25%) to withstand or even some times to excel over the control diet in supporting birds performance is an indication of optimum mixing ratio between pigeon peas and carbohydrates that should precede the commence of fermentation process. In this respect, the explanation of the beneficial effect of the fermented pigeon peas with low levels of carbohydrates (20% sorghum grain or 25% sugar)is either due to nutrients complementary of the combined two feed and/or through the aspect of fermentation process facilitation. The first aspect of the preceded elucidation is in line the earlier view of Bressni (1993) that fermentation of food legumes with cereal grains was proven to have a positive complementary effect in improving nutritional quality of the fermented products.

With regards to the interactive dis–relationship in birds response to the dietary treatments during the starting phase versus the finishing stage, the in capabilities of the dietary treatments containing fermented pigeon peas with high levels of carbohydrates (40–80%) during finishing period to support birds performance similar to that of the control diet or their counterparts with low levels of carbohydrates (20–025%) could be justified in accord to the following understandings. First, the significant drop in feed intake with subsequent growth retardation during the finishing stage rather than the starting phase of the birds fed on dietary treatments containing fermented pigeon peas with high levels of carbohydrates could be attributed to the relative absence of taste sensitivity in younger birds. There is evidence that development of salivary glands which took place at late age is documented to play role in solubilization a number of chemicals in the feed which once have been transferred into solution phase can be detected by the developed taste buds. Furthermore, there is an evidence that development completion of the salivary glands will enhance feed intake and digestion via their lubricant, enzymatic activities and buffering capacities. The second view of interpreting the significant decrease (P<0.05) in performance of birds that were subjected to dietary treatments containing the fermented pigeon peas with high levels of added carbohydrates (40–80%) during the finishing period rather than the starting phase could be attributed to the changes in preservation power of the fermented products which may be deteriorated by the time factor. In this sense, Steinkraus (1996) had pointed out that fermentation process plays five roles in enhancing food quality among which is the preservation of substantial amounts of food through its lactic acid, alcoholic, acetic acid in addition to alkaline or high salt fermentation processes.

CONCLUSION AND RECOMMENDATIONS

On the basis of preliminary results of this study, one can conclude that although the fermentation process of pigeon peas (C. cajan) with low levels of carbohydrates (20% sorghum grains or 25% sugar) seems to be necessary for improvement of nutritional quality of the grain legume, pigeon peas, addition of carbohydrates at higher levels (40–80%) are often detrimental to efficient growth and feed utilization efficiency of broiler chicks.

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