Blood collection for haematological and serum evaluation of starter broilers was carried out at third week of the feeding trial when the birds were four weeks old. Birds were selected per replicate and blood samples collected through vein puncture of the wing vein using sterilized disposable needle after sterilizing the site with methylated spirit. Blood samples were delivered into a set of clean labeled bottles containing ethylene diamine tetra acetic acid (EDTA) for haematological evaluation. While samples for serum biochemical evaluation were delivered into bottles free from EDTA. Haematological parameters evaluated were packed cell volume (PCV), red blood cell (RBC), haemoglobin (Hb), mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, white blood cell, neutrophil and lymphocyte. Serum metabolites evaluated include total protein, albumin, creatinine, aspartate amino transferase (AST), alanine amino transferase (ALT) and glucose.

Proximate Analysis of Test Ingredient and Diets

Samples of the toasted bambaranut waste (TBNW) and diets were analyzed for their proximate composition according to the method of A.O.A.C. (2010).

Data Analysis

All data obtained were subjected to Analysis of Variance (ANOVA) at 5% probability level while Duncan’s Multiple Range Test was used to separate the mean that were significantly different, using SPSS version 2.0 computer statistical software package.

RESULTS AND DISCUSSION

Results

Proximate Composition of Toasted Bambaranut Waste and Experimental Diets of Broilers

The proximate composition of TBNW fed to broilers is shown in Table 2. Determined proximate composition of TBNW and energy showed dry matter (DM) 89.71%, crude protein (CP) 14.99%, crude fibre (CF) 2.00%, ether extract (EE) 3.58%, nitrogen-free extract (NFE) 67.02%, ash 2.12% and energy 3040.35kcal/kgME.

Table 2: Proximate Composition of Toasted Bambaranut Waste

*Metabolizable energy (37 x %CP+ 81 x %EE+ 35.5 x %NFE) (Pauzenga, 1985)

The proximate composition of the experimental diets is presented in Table 3. DM ranged from 89.04 - 89.49%, CP 22.01-25.01%, CF 5.20-5.84%, EE 5.10-6.21%, ash 6.56-7.49%, NFE 46.56-48.11% and energy 2948.95-2964.80 Kcal/KgME.

Table 3: Proximate Composition of Experimental Diets for Starter Broilers

*Metabolizable energy (37 x %CP+ 81 x %EE+ 35.5 x %NFE) (Pauzenga, 1985)

Performance of Starter Broilers Fed Toasted Bambaranut Waste Diets

Table 4 shows the effect of dietary TBNW on performance of starter broilers. There were significant differences (P < 0.05) among treatments in final body weight (FBW), daily weight gain (DWG) and daily feed intake (DFI) but there was no significant difference (P > 0.05) in feed conversion ratio (FCR). Chicks fed the control diet had significantly (P < 0.05) higher FBW and DWG than chicks fed 5, 10 and 20% TBNW diets. Chicks fed 20% TBNW diet had poorest (P > 0.05) FCR than chicks fed other diets. The final body weight ranged between 468.92 and 537.58g, daily weight gain 18.55-21.38g, daily feed intake 44.24-49.52g and feed conversion ratio 2.34 - 2.79.

Table 4: Performance of Starter Broilers Fed Toasted Bambaranut Waste diet

abcd: means on the same row with different letters are significantly different (P<0.05). * = significant, SEM = Standard error of mean, LOS = Level of significance, NS =Not Significant (P>0.05)

Haematology of Starter Broilers Fed Toasted Bambaranut Waste Diets

Haematological profile of the experimental birds is shown in Table 5. There was no significant difference (P>0.05) among treatments in packed cell volume (PCV). However, haemoglobin (Hb), white blood cell (WBC), red blood cell (RBC), mean corpuscular, haemoglobin concentration (MCHC), mean corpuscular haemoglobin (MCH) neutrophils and lymphocytes values of the birds were significantly different (P<0.05). PCV ranged from 30.00 to 31.33%, Hb ranged from 10.27 to 11.60g/dl, WBC ranged 21.07 to 22.86x10⁹/l, RBC ranged from 2.30 to 2.86x10¹²/l, MCV ranged from 107.33 to 122.67fl, MCHC ranged from 33.43 to 35.10%, MCH ranged from 36.00 to 41.50pg, neutrophils ranged from 4.00 to 6.67% and lymphocytes ranged from 93.33 to 96.00%.

Table 5: Haematology of Starter Broilers Fed Toasted Bambaranut Waste Diets

abcd: means on the same row with different letters are significantly different (P<0.05). * = significant, SEM = Standard error of mean, LOS = Level of significance, NS =Not Significant (P>0.05). MCHC-Mean corpuscular haemoglobin concentration, MCH-Mean corpuscular haemoglobin

Serum Biochemistry of Starter Broilers Fed Toasted Bambaranut Waste Diets

Table 6 shows the serum biochemical indices. There were significant differences (P < 0.05) among treatments in total protein, globulin, creatinine, glucose, aspartate aminotransferase, alanine aminotransferase but there was no significant difference (P>0.05) in the serum albumin of birds across the treatments. Total protein ranged from 3.20 to 3.77mg/dl, albumin ranged from 2.10 to 2.37g/dl, globulin ranged from 1.30 to 1.67g/dl, creatinine ranged from 0.47 to 0.80mg/dl, glucose ranged from 195.33 to 219.67mg/dl, aspartate aminotransferase ranged from 195.00 to 258.00µ/l, and alanine aminotransferase ranged from 12.40 to 20.50µ/l.

Table 6: Serum Biochemistry of Starter Broilers Fed Toasted Bambaranut Waste Diets

abcd: means on the same row with different letters are significantly different (P<0.05). * = significant, SEM = Standard error of mean, LOS = Level of significance, NS =Not Significant (P>0.05)

DISCUSSION

The observed DM is higher than 88.00% reported by Ani et al., (2012a), but lower than 89.80% reported by Ani et al., (2012a). The observed difference may have resulted from differences in varieties of seeds used and from the processing methods (Ani et al., 2012a). However, the observed dry matter (DM) value of the TBNW used in this study shows that it was well dried to preserve for long. Crude protein (CP) is lower than 18.30% reported by Ani et al., (2012), 17.30% by Anhwamge et al., (2006) and 17.50% by Emendu and Emendu (2014). Consequently, TBNW cannot be used as protein ingredient. The low value of crude fibre shows that TBNW can be digested and readily utilized by monogastrics. The value is lower than 11.30% reported by Amaefule and Osuagwu (2005) and 20% obtained by Ani et al., (2012a). The observed difference may have resulted from differences in processing methods employed. The observed value for ether extract (EE) shows that toasted bambaranut waste (TBNW) contains fat which is a source of energy. Anhwamge and Atoo (2015) opined that high EE content of the plant indicates the seed could provide more heat and energy to the body. The observed value of EE in this study is lower than the value of 5.36% reported by Ani et al., (2012a) and 5.00% reported by Emendu and Emendu (2014). The observed difference might be due to the differences in variety of seeds used, soil type and the method of processing the TBNW (Amaefule and Obioha, 1998; Ani, 2006). The value of ash shows that TBNW has minerals that could maintain oasmotic pressure and regulate the exchange of water and solute within the animal body as well as the essential constituents of the skeletal structure (Aduku, 2004). The observed nitrogen free extract (NFE) shows that TBNW contains enough soluble carbohydrate and other digestible and easily utilizable non-nitrogenous substances that can make the animal easily digest and absorbed the TBNW. The observed NFE is close to 67.84% reported by Emendu and Emendu (2014) and higher than 41.64% reported by Ani et al., (2012a). Anhwamge and Atoo (2015) stated that, high carbohydrate content of plant material indicates it could provide more heat and energy in the body. The estimated metabolizable energy (ME) of the TBNW is an indication that the material had enough energy value needed by animals to grow, maintain their structures and as well respond to their environment. The ME value of TBNW is lower than 3860.4kcal/kgME reported by Emendu and Emendu (2014).

Proximate Composition of Experimental Diets for Starter Broilers

The observed dry matter (DM) values of the experimental diets used in the study may suggest that all diets were low in moisture content which is an indicator of good quality and a key to safe storage (Bala, 1997). NRI (1995) stated that feed with high moisture content, temperature and poor aeration during storage is predisposed to mycotoxins and spoilage which can pose health problems to birds when fed. The observed crude protein (CP) shows that the experimental diets were adequate in protein required for the synthesis of body tissues, physiological molecules and carcass growth and development (Ofori et al., 2019). Ofori et al., (2019) reported that increase in CP of diets results in improvement in weight of birds. The observed crude fibre values are higher than the recommended values for broilers (NRC, 1994; Aduku, 2004). However, they are similar and do not appear to have adverse effect on feed utilization. The observed values of ether extract (EE) show that the diets contain adequate fat which can generate heat and energy for the birds (Anhwamge and Atoo, 2015). Observed values are above the NIS (1989) standard. According to NRC (1994), fat component of poultry diet helps to increase overall energy concentration and in turn improve productivity and feed efficiency. The observed values for ash show that the experimental diets had sufficient minerals (Aduku, 2004), and are higher than the range of 2.50-3.50 % reported by Ani et al., (2012a). The observed difference may be attributed to ingredient combination and method of processing the diets (Amaefule and Obioha, 1998; Ani and Okorie, 2005).

NFE values of the experimental diets are within the range of 40.50-53.31% observed by Ani et al., (2012a). This indicates the presence of soluble carbohydrate that can be converted to energy.

The metabolizable energy (ME) of the experimental diets shows that the diets contain adequate energy needed by the birds to grow, maintain their structure and as well as respond to their environment conditions. The observed ME values are below the range recommended by NRC (1994). However, they are similar for all the diets.

Performance of Starter Broilers Fed Toasted Bambaranut Waste Diets

Birds fed the control diet had the highest final body weight (FBW). FBW appear to decline as the level of TBNW increased in the diets. Birds on the control diet had the best daily weight gain (DWG) while 20% TBNW had the poorest. The pattern of daily feed intake was not definite. However, observed values may suggest that the feed intake was adequate. Consequently, TBNW in diet did not depress intake. The observed values for feed conversion ratio did not indicate that TBNW in the diets compromised feed utilization for growth.

Haematology of Starter Broilers Fed Toasted Bambaranut Waste Diets

Observed packed cell volume (PCV) across the treatments fall within the normal range of 22 - 35% for healthy chickens reported by Bounous and Stedman (2000), while Peter (2002) reported normal avian PCV to be 40 - 60%. Oyewole et al., (2018a) reported 29.00% - 31.00% for PCV for healthy broilers. Isaac et al., (2013) stated that PCV is involved in the transport of oxygen and absorbed nutrients. On the other hand, Oyawoye and Ogunkunle (1998) reported that PCV value is an index of toxicity in the blood, when low. The PCV values are therefore adequate as these are within reference ranges for healthy birds. Consequently, TBNW did not impair oxygen transport and nutrient absorption. More so, it did not result in toxicity. The observed values of haemoglobin (Hb) of birds fall within the normal range of 7-13g/dl reported by Bounous and Stedman (2000), Ikhimioya et al., (2000) and Oyewole et al., (2018a) for healthy birds. Etim et al., (2014) reported that haemoglobin react with oxygen carried in the blood to form oxyhaemoglobin during respiration. Hence the birds had sufficient vitamins and minerals from all the diets which enabled them to synthesize sufficient haemoglobin which aided oxygen and carbon dioxide transportation in the blood (Oyewole et al., 2018a). The values for white blood cell (WBC) obtained in this study are within the normal range of 12-30x10⁹/l reported by Bounous and Stedman (2000) for healthy birds. WBCs are responsible for defense of the body against infection. They defend the body by phagocytocis against invasion by foreign organisms and produce or at least transport, distribute immune responses (Etim et al., 2014). The values of WBC show that TBNW inclusion in the diets did not compromise immune status of birds. The values of red blood cell (RBC) obtained in this study were within the range of 2.5-3.5x10¹²/l reported by Bounous and Stedman (2000) for healthy birds. RBCs (erythrocytes) serve as carrier of haemoglobin (Etim et al., 2014), and are involved in the transport of oxygen and carbondioxide in the body. Hence, RBC count observed in this study implies that adequate levels of oxygen were carried to the tissues as well as carbon dioxide returned to the lungs (Soetan et al., 2013; Isaac et al., 2013 and Etim et al., 2014; Ani and Omeje, 2011). A low level of RBC may be due to anaemia (Peter, 2002), the birds were not anaemic. The observed mean corpuscular volume (MCV) values are within the ranges of 90-140fl and 132.27-134.87fl reported by Bounous and Stedman (2000) and Oyewole et al., (2018a) respectively, for healthy broilers. Thus the values of MCV obtained in this study indicated that the TBNW diets where adequate in terms of vitamin and minerals. Hence, the birds were healthy, not anaemic and capable of withstanding stress (Maxwell, 1993). Mean corpuscular haemoglobin concentration (MCHC) were higher than the normal ranges of 32.4-33.21% and 29.80-31.63g/dl reported by Wikivet (2012) and Oyewole et al., (2018) respectively, but fall with the range reported by Mitruka and Rawnsley (1977) for healthy birds. Aster (2004) advanced that low level of MCHC is an indication of anaemia. Oyawoye and Ogunkunle (1998) opined that MCHC is useful in monitoring feed toxicity especially with feed constituents that affect the blood as well as the health status of farm animals. Thus the values of MCHC obtained in this study may indicate that the TBNW diets were adequately utilized by the birds and posed no problem to them. The values of MCH obtained in this study were within the normal range for healthy birds. (Bounous and Stedman (2000) recorded the range of 33-47pg while Oyewole et al., (2018a) reported 40.10pg-41.87pg. Lower levels of MCH may indicate anaemia. Observed MCH values may indicate that the bone marrow of bird was functioning normally, hence the absence of macrocytic and hypochronic anaemia. The observed values for neutrophils are within the normal range for healthy birds. Since the main function of neutrophils is phagocytosis, the values of neutrophils of birds on TBNW diet are adequate for ingestion and destruction of foreign particles (Maxwell and Robertson, 1998). The observed values for lymphocytes are within the normal range for healthy birds (Oyewole et al., 2018b). The values of lymphocytes in this study therefore show that TBNW inclusion in broiler diets did not have negative effect on the immune response of birds.

Serum Biochemistry of Starter Broilers Fed Toasted Bambaranut Waste Diets

The total protein values indicate that birds fed the control diet had the highest value which declined with the inclusion of TBNW. Nonetheless, all values observed are within the normal range of 2.5-4.5g/dl observed by Thrall (2007); 3-5g/dl for birds reported by Elliot et al., (1974); 3.0-4.9 by Meluzzi et al., (1992); 3.5-5.5gdl reported by Peter (2002) and 3.97-4.80g/dl for starter broilers fed cashew pulp meal based diets (Oyewole et al., 2017). Peter (2002) stated that low protein level is an indication of poor diet. Serum albumin values are all within the normal albumin range of 1.17-2.74g/dl reported by Meluzzi et al., (1992) and 1.73g/dl to 2.37g/dl (Oyewole et al., 2017). Globulin values are within the normal range of 1.15-1.53g/dl reported by Chicken-Biochemistry (2016) and the 0.5-1.8g/dl reported by Thrall (2007). The creatinine level of birds fed the control diet was lower than the creatinine levels of birds fed the TBNW diets. All the values of creatinine reported are within the normal reference range of 0.5-1.5mg/dl (Levey et al., 2006). Creatinine concentrations lower than the reference range for young animals are usually caused by decreased muscle mass due to muscle atrophy arising from starvation (Laskin et al., 2014). Younger and older broilers have low levels of creatinine (Sandhu et al., 1998). An elevated level of creatinine usually indicates a muscular condition due to poor quality of protein in the diets. This was not so with the birds in this study. Serum glucose values fall within the range reported by Sandhu et al., (1998) and Peter (2002) for broilers. According to Wasserman (2009), a persistent elevation in blood glucose levels leads to glucose toxicity which contributes to cell dysfunction. This may indicate that TBNW may not have had adverse effect on the birds.

Aspartate aminotransferase (AST) values in this study reveal that the birds responded differently to the TBNW diets. All AST values are within the range of 70-220µ/l reported by Meluzzi et al., (1992) and 102.33-135.67µ/l (Oyewole et al., 2017). Serum values greater than 350µ/l are considered abnormal and are often indicators of liver disease. The observed values showed that the TBNW inclusion in the diets did not cause liver damage to birds. Hence, TBNW is safe when fed to animals. Alanine aminotransferase (ALT) values in this study are above 4.00-7.33µ/l (Oyewole et al., 2017) but close to 18.99-20.06 µ/l reported by Abdel-Fattah et al., (2008) for healthy birds. The observed values showed that the TBNW inclusion in the diets did not cause liver damage to birds. Hence, TBNW is safe when fed to starter broilers.

CONCLUSION

Inclusion of TBNW in the diets resulted in poorer performance indices of starter broilers. However, broilers fed 10% inclusion had similar weight with those on the control. Hence starter broilers can be fed up to 10% TBNW in the diet. The feeding trial also revealed that TBNW is safe for starter broilers as no deleterious effects were observed in the blood parameters of the birds even at 20% inclusion.

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