Response of Broiler Chickens Fed Boiled Red Kidney Bean ( Phaseolus vulgaris ) Seed Meal as Protein feedstuff on Haematology and Serum Biochemical indices

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INTRODUCTION
The main limitation to expansion of poultry industry is the availability of adequate supplies of needed ingridients at reasonable prices (Ani and Okorie, 2005;Babatunde and Hamzat (2005). Mean while, poultry products have been considered to be one of the options in Nigeria for reducing the incidence of malnutrition, particularly protein deficiency in diets of the populace (Amaefule et al., 2009). The exorbitant cost of commercial poultry feeds had been identified to be largely due to high cost of Conventional feedstuffs. Feed account for about 70% of total cost of production (Ademola and Farinu, 2006). A high cost of feed is due mainly to competetion between man and Livestock for grains and conventional sources of plant (Soya beans, groundnut seed) and hence animal proteins (Emenalon, 2004). The most promising way to solve the problem of competetion between man and animal for plant protein is to identify cheaper and available feed stuff that have less or no industrial use that can meet nutritional requirement of poultry with or without processing (Akinmutimi, 2001). One of such seeds that have potential of being used in poultry feeds is the Red kidney Bean seed (Phaseolus vulgaris). The kidney bean (Phaseolus vulgaris) also known as common bean, haricot bean, navy bean, dry bean, snap bean, French bean and pinto bean are native of Central America (Olomu, 2011). The bean is an herbaceous annual plant grown worldwide for its edible fruit, either the dry seed or the unripe fruit both of which are referred to as beans (Philips and Rix, 1993). Kidney bean is a leguminous crop grown in different parts of the world. Several varieties of this crop are currently cultivated in fairly large quantities in Mangu and Bokkos areas of Jos, Plateau State, Nigeria and are worth investigating for use as a possible source of protein ingredient in poultry feed production.
However, like other grain legumes, its usefulness as a feed ingredient for monogastric animals is limited due to the presence of some anti-nutritional substances in feedstuffs which has been associated with growth depression and pancreatic hypertrophy in many monogastric animal species (Birk, 1988;Olomu, 2011). However, it has been established that heat treatment and other processing methods exert beneficial efforts on the seeds of the grain legumes by destroying the anti-nutritional factors inherent in them (Balogun et al., 2001). Thus boiling tends to increase palatability and protein intake (Goshit, 2015). Results obtained from the processed methods of red kidney beans (roasting, cooking, salting, and chemical treated) improved the amino acid availability. Against this background the present study focused on the use of Red kidney bean seeds boiled in alkaline salt (potassium bicarbonate) Solution as protein feedstuff for broiler chickens.

Location of Experiment
The study was carried out at the Poultry Research Farm of the Department of Animal Science and Range Management of Modibbo Adama University, Yola, Girei Local Government Area of Adamawa State, Nigeria. Farm lies between latitude 9 0 and 11 0 N of the equator and longitude 10 0 and 14 0 E of the Greenwich meridian. Adamawa shares its boundaries with Taraba state to the South and West, Gombe state to the North-West and Borno state to the North. The state has tropical climate with distinct dry and wet seasons. It has an average minimum and maximum temperatures of 18 and 40 0 C and relative humidity of 20 and 80% Adamawa state has an international boundary with the Cameroon Republic along its eastern boarder (Adebayo, 1999).

Source of Red Kidney Bean (Phaseolus Vulgaris) and Raw Materials
The red kidney bean seeds were purchased from Mangu market in Mangu Local Government Area of Plateau State. Other ingridients like Alkaline salt, Potassium bicarbonate (KCO2) were purchased from Jimeta market. The seeds were subjected to the following processing methods before using it for the feeding trial.

Processing of Red Kidney bean seeds
Cold clean water was brought to boiling point (100 0 C) in a 200 litres capacity half cut drum; a modified method of boiling pigeon pea seeds was adopted. 100g of alkaline salt, potassium bicarbonate (KCO2) was dissolved to produce an aqueous solution. Raw red kidney bean seeds was poured into the boiling water and covered. After 60 minutes, the content was drained off and boiled seeds were sundried for 4 days, before milling and then used to formulate Boiled Red Kidney Bean Seed Meal (BRKBSM) diets.

Experimental Diets and Chemical Analysis
Raw and boiled red kidney bean seeds (Table 1) were analysed for proximate composition before used in formulating the experimental diets. Five experimental broiler Starter and finisher diets were formulated as in Tables 2 and 3. Diet 1 (T1) which served as Treatment Control contained 0% Boiled Red kidney bean seed meal while treatments 2, 3, 4 and 5 contained Boiled Red Kidney Bean Seed Meal (BRKBSM) at 25, 50, 75 and 100 % levels respectively. All diets were formulated to be isocaloric (ME 3137 and 3232 kcal/kg) and isonitrogenous (19 and 22 % CP) at both starter and finisher phases respectively. The Raw and boiled red kidney bean seeds (Table 1), and formulated broiler starter finisher feeds (Table 2 and 3) samples were analyesd for proximate composition according to the standard described by AOAC (2010) as reflected in determined analysis.    (Anak, 2000) white strain broiler chicks were used for the experiment. Birds were weighed and randomly allotted to five dietary treatments of 40 birds per Treatment, 10 birds per replicate. Each treatment was replicated 4 times in a completely randomized design (CRD). Weighed quatities of experimental Diets were supplied to each group every morning. Left-over feeds were deducted from the quantity supplied the next moringing to determine the feed intake. Water was supplied ad libitum throughout the period of the experiment for (8 weeks).

Hematological and Serum Biochemical Studies
On the 56 th day of the feeding trial, the birds were starved overnight, four birds per treatment were randomly picked and each was bled. This was achieved by puncturing the web vein with a 5 ml gauge syringe and needle, transfering the blood into labeled sterile universal bottle containing 1.0mg/ml ethylene diamine tetracetate acid (EDTA) as anti-coagulant. This was used to determine the hematological components according to the method described by Ajagbonna et al. (1999) and Uko et al. (2000). Another 5 ml was collected into a labeled sterile sample bottles without coagulant and was used to determine the biochemical components (Ajagbonna et al., 1999;Uko et al., 2000). Parameters determined include Packed Cell Volume (PVC), Hemoglobin Concentration (Hb), Red Blood Cell (RBC) and White Blood Cell (WBC), Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin Concentration (MCHC) and Mean Corpuscular Haemoglobin (MCH). The biochemical indices included the level of glucose, total protein, globulin, albumin, urea, creatinine and cholesterol.

Statistical Analysis
All data generated were subjected to One-Way Analysis of Variance (ANOVA) as described by Steel and Torrie (1980). And treatment means were compared using Duncan Multiple Range Test (DMRT) (Duncan, 1955).

Haematology
Parameters for haematological assay were not significantly affected by the treatment diets (P>0.05) except MCV (P<0.01) and MCHC (p<0.05) respectively (Table 4). Values obtained for these parametres fell within the range established by Mitruka and Rawnsley (1977). Similarities in the values of heamoglobin (Hb) among birds irrespective of inclusion levels implies that diets were adequate in critical nutrients such as; iron, copper, vitamins and amino acids. Similarly, Sainsbury (1983) reported that haemoglobin measures the ability of an animal to withstand some level of respiratory strees. RBC values fell within the normal range reported by Roberts et al., 2003. However, it did not agree with the work of Adejinmi et al. (2000) who reported inconsistent RBC values for broilers. The present study suggests that blood of birds had appreciable oxygen carrying capacity which also indicates nutrient transport by blood was not impaired by feeding the test diets. Iheukwemere (2008) observed strong influence of diets in heamoglobin traits with packed cell volume (PCV) as being strong indicators of nutrition status of the animals. White blood cell values for birds fed BRKBSM followed the same pattern with RBC values. Values obtained in this study fell within normal limits for chickens (Ross et al., 1979). Esonu et al. (2001) reported that quality of haematological indices is always a reflection of animals responsiveness to both internal and external enviroments which includes feed and feeding. However, all heamatological values fell wothin the range for chickens (Maxwell et al., 1990;CCAC, 1993).

Serum Biochemical Indices
The results of the biochemical estimation of serum total protein, blood urea nitrogen, blood glucose, albumin, globulin, creatinine and cholesterol values (Table 5) showed that there were no significant (P>0.05) difference. Bamgbose et al. (2003) reported that total protein and albumin are indicators of protein reserve as extra source of energy to the birds (Mayes, 1996). Globulin values obtained for all the test diets could be due to the adequate protein in the treatment diets as was observed by Awojobi and Opiah (2000) that the higher the level of Globulin the greater the ability to fight infections. Urea and creatinine values indicated no obvious muscular wastage due to protein adequacy.

CONCLUSION
The over all result indicated that Boiled Red Kidney Bean Seed meal is a satisfactory protein source which can be included in the broiler chickens diets up to 50% without any deleterious effect on the growth performance and heamatological and serum biochemical indices of broiler chickens. Hence, 50% inclusion level of Boiled Red Kidney Bean Seed Meal could be recommenmed for broiler diets.

Conflit of Interest
The authors declare no conflict of interest that may affect the outcome of the study in any way.