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Laying performance of Japanese quail fed graded levels of neem (Azadirachta indica) kernel meal incorporated diets
Animal Feed Science and Technology 88 (2000) 113±120
Short communication
Laying performance of Japanese quail fed graded levels of neem (Azadirachta indica) kernel meal incorporated diets A.V. Elangovana, S.V.S. Vermaa,*, V.R.B. Sastryb, S.D. Singhb a
Division of Avian Nutrition and Feed Technology, Central Avian Research Institute, Izatnagar 243122, India b Indian Veterinary Research Institute, Izatnagar 243122, India Received 3 November 1999; received in revised form 27 April 2000; accepted 25 July 2000
Abstract The response of laying Japanese quails to graded amounts of dietary neem kernel meal (NKM) added for part of soybean meal and deoiled rice bran was investigated in the present study. The NKM was incorporated at 0, 50, 75 or 100 g kgÿ1 in a standard layer diet, and each of the four test diets were offered ad libitum as mash to 12 birds housed individually in laying cages from 7 to 20 weeks of age. The birds in different dietary groups had earlier received the respective quail starter±grower diets containing graded amounts of NKM as in the layer quail diets. Observations on egg production of quails, feed intake, blood biochemicals and pathomorphology of vital organs were made. Results on feed intake revealed insigni®cant differences amongst the groups and almost similar egg production and egg quality. However, quails fed the NKM diets tended to utilize feed less ef®ciently (P < 0:01) as compared to those fed the control diet. Except for signi®cantly high (P < 0:01) ALP activity in quails fed the NKM diet, no signi®cant (P < 0:05) differences in haematological (Hb, TEC, TLC), serum biochemicals (glucose, protein, cholesterol), and the AST, ALT activities were noticed. Regression analysis of data showed a decreasing trend of egg production with poor feed:egg and an increased TLC, protein and ALP as the dietary level of NKM was increased. Histopathology of vital organs of quails fed the NKM diets showed massive fatty in®ltration in liver and degenerative changes in the kidney tissues. From the results of the study it may be concluded that feeding NKM to Japanese quails from day 1 to 20 weeks of age supported normal feed intake, egg production and egg quality but, however, the ingredient on long-term feeding induced pathological changes in liver and kidney tissues. # 2000 Elsevier Science B.V. All rights reserved. Keywords: Quails; Neem kernel meal; Egg production; Haematology; Histopathology
*
Corresponding author. Tel.: 91-581-447223; fax: 91-581-447321. E-mail address: [email protected] (S.V.S. Verma). 0377-8401/00/$ ± see front matter # 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 7 7 - 8 4 0 1 ( 0 0 ) 0 0 1 8 4 - X
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A.V. Elangovan et al. / Animal Feed Science and Technology 88 (2000) 113±120
1. Introduction The neem (Azadirachta indica) kernel meal (NKM) obtained on solvent extraction of neem seeds is available in sizable quantities of about 0.7 million MT per annum in India (Singh, 1993). However, despite of its high protein content of 300±400 g kgÿ1 the meal could not earn favour as a good livestock feed due to primarily to its pungent smell and bitter taste imparted by the presence of toxic triterpenoids; azadirone, nimbin, nimbidin, salanin, etc., instead utilized for fertilizer-cum-pesticide and also as a ¯y repellent. Several workers (Sadagopan et al., 1981; Verma et al., 1998; Gowda et al., 1998) tried to investigate the feeding value of NKM in the diet of laying hens but with variable success. Similar was the case with chicks (Sadagopan et al., 1982; Reddy and Rao, 1988; Chand, 1987; Jana, 1997). In a short-term study with laying WL hens, Gowda et al. (1998) recommended a safe level of inclusion of NKM 100 g kgÿ1 in diet. Thus the present study was undertaken with laying Japanese quails to examine the effect of inclusion of NKM in diet without adversely affecting their productive performance on long-term feeding. 2. Materials and methods 2.1. Experimental diet and birds One hundred and twenty Japanese quail chicks as hatched were randomly distributed into 12 lots of 10 each and housed in battery cages under similar managerial conditions. A maize soybean meal standard starter±grower mash (D1) with 230 g CP kgÿ1 and 11.27 MJ ME kgÿ1 was made. Three more diets (D2, D3 and D4) containing 50, 75 or 100 g NKM kgÿ1, respectively, for part of soybean meal and deoiled rice bran in diet D1 were made. One of the four diets was offered ad libitum to triplicate lots of chicks from day 1 to 6 weeks of age. At this age, 12 female quails from each of the four dietary groups were picked up and housed in individual laying cages. The respective grower diet were modi®ed in respect of CP and Ca contents to serve as layer diets without altering the NKM contents (Table 1). Each of the four test diets (D1, D2, D3 and D4) were offered ad libitum as mash to the corresponding quails from 7 to 20 weeks of age during which observations on net feed intake and daily egg production were recorded. The NKM and experimental diets were analysed for proximate composition, calcium and phosphorus as per standard procedures (AOAC, 1990). 2.2. Egg quality Ten eggs from each of the four dietary groups laid on ®rst two consecutive days during the 11th, 15th and 19th week of experimental feeding (i.e. 10 3 4 120) were utilized for quality testing in respect of shape index (100 times the ratio of width to length of an egg), albumen index (the ratio of average albumen height to the average of the width and length), yolk index (the ratio of yolk height to the average of the width), internal quality unit (IQU) as per Kondiah et al. (1981), shell weight and shell thickness.
A.V. Elangovan et al. / Animal Feed Science and Technology 88 (2000) 113±120
115
Table 1 Ingredient and nutrient composition of layer diets Dietary group (g NKM kgÿ1)
Attribute
D1 (0)
D2 (50)
D3 (75)
D4 (100)
720 220 60 ±
720 182 48 50
720 162.5 42.5 75
720 145 35 100
Nutrient composition (g or MJ kgÿ1), calculated Crude protein 186.1 Metabolisable energy 11.27 Lysine 9.80 Methionine 3.34
186.2 11.25 9.60 3.42
186.1 11.23 9.49 3.46
186.6 11.22 9.42 3.51
Nutrient composition (g or kgÿ1), determined Crude protein 186.9 Calcium 28.8 Phosphorus 9.33
182.5 28.2 9.35
183.1 29.5 9.41
184.2 30.1 9.12
ÿ1
Ingredient composition (g kg ) Basal mixturea Soybean meal (SBM) Deoiled rice bran (DORB) NKM
a Basal mixture consisted of g kgÿ1 diet: maize, 590; ®sh meal, 50; dicalcium phosphate, 14.3; ground lime stone, 30; oyster shell, 30; common salt, 3.0; trace mineral supplement (per kg diet provided: Mn, 55 mg; I, 12 mg; Fe, 156 mg; Zn, 30 mg; Cu, 4 mg; Co, 0.9 mg), 1.0; vitamin mixture (per kg diet provided: Vit. A, 8250 IU; Vit. D3, 1200 ICU, Vit. K, 1 mg; Vit. E, 40 IU; Vit. B1, 4 mg; Vit. B2, 4 mg; Vit. B12, 4.0 g; niacin, 60 mg; pantothenic acid, 10 mg; choline, 500 mg), 1.7%.
2.3. Organoleptic evaluation Ten eggs from each dietary group laid on the 3rd and 4th day of 11th, 15th and 19th week of feeding (i.e. 10 3 4 120) were evaluated organoleptically on an eight point hedonic scale in respect of colour, ¯avour, taste, smell, odour, texture and overall acceptability by a panel of 10 semitrained judges. For this purpose, eggs were boiled for 5 min and cooled under tap water. 2.4. Haematological and biochemical pro®le For the purpose of haemato-biochemical pro®le, blood samples from six quails per dietary group were drawn into a heparinized (20 IU/ml) glass vials at the end of 20 weeks of experimental feeding for the estimation of haemoglobin (Schalm et al., 1975), total erythrocyte (TEC) and total leucocyte counts (TLC) (Natt and Harrick, 1952). Concomitantly, the serum separated from blood drawn into another set of glass vials without any anticoagulants were stored at ÿ208C for subsequent use to estimate protein (Hiller and Slyke, 1927), cholesterol (Wybenga and Pileggi, 1970), ALT (Reitman and Frankel, 1957), AST (Reitman and Frankel, 1975) and ALP (Kind and King, 1954). The estimation of serum glucose (Cooper and McDaniell, 1970) was carried out immediately.
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2.5. Gross and histopathology Tissues from target organs (intestine, liver, spleen, heart, kidneys and ovaries) were collected from six birds per dietary group at the end of 20 weeks of experimental feeding were examined for gross and histopathological changes to see the effect of feeding NKM. The tissues were preserved in 10% formalin and processed as per standard techniques. 2.6. Statistical analysis The data in respect of various parameters were analysed statistically for treatment effects in a completely randomized design as per Snedecor and Cochran (1967). The data were further subjected to test the linearity among the graded levels of NKM feeding. 3. Results On chemical analysis, each kg of NKM DM was found to contain 382 g CP; 33 g EE; 132 g CF and 118 g total ash. An initial signi®cant (P < 0:01) drop in egg production due to feeding of NKM containing diets to quails improved gradually after 8th week of feeding (Table 2). The overall feed intake and egg production of quails in different groups Table 2 Laying performance of quails fed NKM diets and egg quality Attribute
Dietary group (g NKM kgÿ1) D1 (0)
D2 (50)
D3 (75)
D4 (100)
SEM
Production performance (7±20 weeks of age) Egg production, hen day (%) 7±8 weeks 82.0 7±20 weeks 88.9 Feed intake, 7±20 wk (g/b/d) 27.0 Egg weight (g) 11.6 Feed:egg 2.62 Live weight change 7±20 weeks (g/b) 26.2
62.5 85.3 28.0 11.5 2.86 31.0
55.9 85.8 26.3 10.7 2.91 25.6
57.5 83.8 27.3 11.1 3.00 23.6
6.49 1.72 0.68 0.32 0.062 4.16
Egg quality characteristics Shape index Shell thickness (mm) Shell weight (% of egg weight) Albumen index IQU Yolk index
77.9 0.193 10.5 0.129 62.3 0.478
78.0 0.203 10.5 0.131 62.7 0.467
77.6 0.193 10.4 0.130 63.1 0.471
77.7 0.200 10.4 0.133 63.7 0.471
0.98 0.0143 0.41 0.0063 1.21 0.0125
7.0 7.0 6.8 6.7 6.9 7.0
6.8 6.7 6.7 6.7 6.6 6.9
6.6 6.6 6.9 6.8 6.6 6.8
6.8 6.8 6.7 6.7 6.7 6.9
Organoleptic evaluation of boiled eggs Colour Flavour Taste Smell Texture Overall acceptability
0.33 0.41 0.38 0.29 0.35 0.28
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Table 3 Haemato-biochemical pro®le of quails fed NKM diets Attribute
Hb (g/dl) TEC (106 mmÿ3) TLC (103 mmÿ3) Glucose (mg/dl) Protein (g/dl) Cholesterol (mg/dl) ALT (IU/L) AST (IU/L) ALP (IU/L)
Dietary group (g NKM kgÿ1 diet) D1 (0)
D2 (50)
D3 (75)
D4 (100)
SEM
11.2 3.58 22.1 242 3.78 268 18.6 85.3 39.5
10.9 3.42 24.1 231 3.86 269 17.6 86.2 49.2
10.5 3.39 24.0 242 4.01 275 16.8 83.2 53.8
10.0 3.55 24.6 225 4.09 252 18.5 87.5 55.6
1.23 0.443 1.56 17.3 0.020 15.5 0.84 4.37 2.76
for the 20-week period did not differ signi®cantly (P > 0:05) due to dietary treatments. However, quails fed NKM containing diets required signi®cantly (P < 0:01) more feed for producing a unit egg mass. The pooled means at 11th, 15th and 19th week of experimental period for egg quality characteristics in terms of shape index, shell thickness, shell weight, albumen index, IQU and yolk index revealed no signi®cant (P > 0:05) difference assignable to dietary treatments (Table 2). Data on organoleptic evaluation of boiled eggs also did not show up any untoward characteristics in terms of colour, ¯avour, taste, smell, texture or the overall acceptability of the product that could be assignable to dietary treatments. The haematological and biochemical characteristics evaluated in terms of haemoglobin, TEC, TLC, glucose, protein and cholesterol revealed no signi®cant (P > 0:05) differences attributable to the feeding of NKM to quails (Table 3). Of the activity of various enzymes, the activity of ALP was found signi®cantly (P < 0:01) higher for quails fed the NKM diets than those of quails receiving the reference diet. The activities of ALT and AST, however, remained unaltered. Quails in all of the four treatment groups were in the state of positive weight gain and also no mortality assignable to the feeding of NKM were recorded during the experimental period. The linear regression equation of laying performance of quails and the haematobiochemical pro®le has been presented in Tables 4 and 5. The percent egg production of quails at 7±8 weeks and 7±20 weeks of age showed a signi®cant decrease (P < 0:065 and P < 0:057, respectively) with poorer feed:egg (P < 0:011) at higher levels of NKM in diet. Among the egg quality characteristics, the IQU showed a signi®cant increase (P < 0:036) as the level of NKM increased. As for haematobiochemicals, there was a decrease in haemoglobin (P < 0:042) and an increase in TLC (P < 0:058), protein (P < 0:035), ALP (P < 0:012) as the level of NKM was increased. Rest of all the parameters did not reveal any signi®cant linearity with graded levels of NKM in diet. The gross pathological examination of liver of quails fed diets D2±D4 containing NKM revealed slight enlargement with pale and greyish discolouration with petechieal haemorrhages, whereas the examination of kidney showed mild to moderate degree of
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Table 4 Linear regression equations of laying performance of quails fed NKM diets and egg quality r2
Significance (P)
Production performance (7±20 weeks of age) Egg production, hen day (%) 7±8 weeks Y 79:28 ÿ 0:26X 7±20 weeks Y 88:61 ÿ 0:047X Feed intake, 7±20 weeks (g/b/d) Y 27:21 ÿ 0:001X Egg weight (g) Y 11:62 ÿ 0:007X Feed:egg Y 2:636 0:0038X Live weight change 7±20 weeks (g/b) Y 28:19 ÿ 0:028X
0.87 0.89 0.005 0.52 0.98 0.15
0.065 0.057 0.931 0.276 0.011 0.614
Egg quality characteristics Shape index Shell thickness (mm) Shell weight (% of egg weight) Albumen index IQU Yolk index
Y Y Y Y Y Y
77:95 ÿ 0:0027X 0:1947 0:00004X 10:51 ÿ 0:0011X 0:129 0:0003X 62:19 0:0135X 0:476 ÿ 0:00007X
0.41 0.14 0.71 0.69 0.93 0.40
0.359 0.624 0.155 0.171 0.036 0.371
Organoleptic evaluation of boiled eggs Colour Flavour Taste Smell Texture Overall acceptability
Y Y Y Y Y Y
6:93 ÿ 0:0022X 6:91 ÿ 0:0024X 6:84 ÿ 0:0006X 6:68 0:0005X 6:86 ÿ 0:0026X 6:98 ÿ 0:0009X
0.45 0.41 0.049 0.22 0.66 0.31
0.330 0.358 0.779 0.532 0.188 0.447
Attribute
Linear equation
congestion. Microscopic examination showed a varied degree of degenerative changes in liver paranchyma with extensive fatty and mononuclear in®ltration even in those of quails fed the low level of 50 g kgÿ1 NKM in diet. Similarly, mild to moderate degree of degenerative changes were observed in the kidney tissues. The gross and microscopic examination of other vital organs revealed a normal picture of the tissues with no untoward changes assignable to NKM feeding.
Table 5 Linear regression equations of haemato-biochemical pro®le of quails fed NKM diets Attribute
Linear equation
r2
Significance (P)
Hb (g/dl) TEC (106 mmÿ3) TLC (103 mmÿ3) Glucose (mg/dl) Protein (g/dl) Cholesterol (mg/dl) ALT (IU/L) AST (IU/L) ALP (IU/L)
Y Y Y Y Y Y Y Y Y
0.92 0.10 0.89 0.39 0.93 0.21 0.09 0.05 0.98
0.042 0.678 0.058 0.376 0.035 0.543 0.706 0.779 0.012
11:31 ÿ 0:012X 3:52 ÿ 0:0007X 22:34 0:024X 241:9 ÿ 0:12X 3:76 0:003X 271:9 ÿ 0:105X 18:20 ÿ 0:0058X 85:02 0:009X 40:15 0:17X
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4. Discussion The initial egg production of quails fed the NKM containing diets appeared to be signi®cantly (P < 0:01) lower as compared to those fed the soybean meal containing reference diet and which may have been due to delayed sexual maturity. Such an effect was, however, not indicated by earlier workers (Verma et al., 1998; Gowda et al., 1998) in WL hens probably because the NKM was not introduced in diet from day one as has been done in the present study. However, the adverse effects of NKM feeding to quails were not re¯ected in the overall egg production which was found comparable among the different dietary groups as has been reported by earlier workers with WL hens (Sadagopan et al., 1981; Verma et al., 1998; Gowda et al., 1998). The overall feed intake among quails on different diets was also found comparable as has been experienced by earlier workers in WL hens fed NKM up to 100 g kgÿ1. But in the study of Gowda et al. (1998), they observed a lowered egg production and feed intake by WL hens fed NKM in diet at levels more than 100 g kgÿ1. However, in the present study, a decreasing trend was observed both at initial as well as overall egg production with graded levels of NKM in diet on regression analysis. In spite of similar egg production rate and feed intake of quails in different dietary groups, feed required for unit egg production was higher (P < 0:01) in those birds fed NKM diet than those on reference diet and such observations corroborate to the ®ndings of Sadagopan et al. (1981) in WL hens. On the contrary, earlier works of Verma et al. (1998) and Gowda et al. (1998) did not observe any signi®cant effect on feed:egg ratio in WL hens fed NKM up to 100 g kgÿ1 diet. The mean egg weight and body weight changes of quails in different dietary groups being similar and that all the birds showed positive growth. The egg quality characteristics as well as the organoleptic evaluation of boiled quail eggs were almost similar for all the dietary groups as also observed by earlier workers (Verma et al., 1998; Gowda et al., 1998) while employing WL hens for their study indicating that the bitter taste of triterpenoids were not transferred to the eggs. Feeding NKM to quails appeared to have not in¯uenced the haemato-biochemical pro®le except for a higher (P < 0:01) ALP activity in quails as compared to those fed the reference diet. Further, a increasing trend of TLC, protein and ALP on regression analysis provided a supportive evidence to the pathological alterations observed in some of the vital organs examined particularly the liver and kidney of the NKM fed quails. The absence of noticeable pathological lesions in WL hens fed 100 g NKM kgÿ1 diet for a 12 weeks period from 25 to 37 weeks of age (Gowda et al., 1998) could, therefore, be argued in terms of a relatively shorter feeding period. Moreover, the introduction of NKM in diet of quails right from day one of age may have accentuated the metabolic changes leading to signi®cant pathological lesions. Thus, it could be concluded that basing on the pathological changes observed in the vital organs, the overall use of NKM at any signi®cant level in the diet of quails may not be very bene®cial. References AOAC, 1990. Of®cial Methods of Analysis, 15th Edition. Association of Of®cial Analytical Chemists. Washington, DC.
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Chand, S., 1987. Nutritional evaluation of neem seed meal in chicks. Ph.D. Thesis. Rohilkhand University, Bareilly, India. Cooper, G.R., McDaniell, V., 1970. o-Toluidine method of blood glucose estimation. In: McDonald, R.P. (Ed.), Standard Methods of Clinical Chemistry, Vol. b. Academic Press, New York, p. 159. Gowda, S.K., Verma, S.V.S., Elangovan, A.V., Singh, S.D., 1998. Neem (Azadirachta indica) kernel meal in the diet of White Leghorn layers. Br. Poult. Sci. 39, 648±652. Hiller, M., Slyke, V., 1927. Colorimetric determination of proteins. J. Clin. Invest. 4, 235. Jana, D.K., 1997. Effect of chemical treatment of neem kernel meal on bioavailability of energy for poultry. M.V.Sc. Thesis. IVRI, Izatnagar, India. Kind, P.R.M., King, E.J., 1954. Serum acid and alkaline phosphatase activity. J. Clin. Pathol. 7, 322. Kondiah, N., Panda, B., Singhal, R.A., 1981. Internal egg quality measures for quail eggs. Indian J. Anim. Sci. 53, 1261±1264. Natt, M.P., Harrick, C.A., 1952. A new blood diluent for counting the erythrocyte and leucocyte of chickens. Poult. Sci. 31, 735±738. Reddy, V.R., Rao, P.V., 1988. Utilization of undecorticated expeller processed or solvent extracted neem cake in chicks. Indian J. Anim. Sci. 58, 835±839. Reitman, S., Frankel, S., 1957. A colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic transaminase. Am. J. Clin. Pathol. 28, 56. Sadagopan, V.R., Johri, T.S., Reddy, V.R., 1981. Feeding value of neem seed meal in broiler and layer diet. Indian Poult. Gazette 65, 136±142. Sadagopan, V.R., Johri, T.S., Reddy, V.R., Panda, B.K., 1982. Feeding value of neem seed meal for starter chicks. Indian Vet. J. 59, 462±465. Schalm, O.W., Jain, N.C., Caroll, E.J., 1975. Veterinary Haematology, 3rd Edition. Lea and Febiger, Philadelphia. Singh, K., 1993. Livestock production and health. In: Neem Research and Development, Publication No. 3. Society of Pesticide Science, India, pp. 187±198. Snedecor, G.W., Cochran, W.G., 1967. Statistical Methods, 6 Edition. Iowa State University Press, Ames, IA, USA. Verma, S.V.S., Gowda, S.K., Elangovan, A.V., 1998. Response of single comb White Leghorn layers to dietary inclusion of raw or alkali treated neem (Azadirachta indica) kernel meal. Anim. Feed Sci. Technol. 76, 169±175. Wybenga, D.R., Pileggi, V.J., 1970. Estimation of cholesterol. Clin. Chem. 16, 980.
Short communication
Laying performance of Japanese quail fed graded levels of neem (Azadirachta indica) kernel meal incorporated diets A.V. Elangovana, S.V.S. Vermaa,*, V.R.B. Sastryb, S.D. Singhb a
Division of Avian Nutrition and Feed Technology, Central Avian Research Institute, Izatnagar 243122, India b Indian Veterinary Research Institute, Izatnagar 243122, India Received 3 November 1999; received in revised form 27 April 2000; accepted 25 July 2000
Abstract The response of laying Japanese quails to graded amounts of dietary neem kernel meal (NKM) added for part of soybean meal and deoiled rice bran was investigated in the present study. The NKM was incorporated at 0, 50, 75 or 100 g kgÿ1 in a standard layer diet, and each of the four test diets were offered ad libitum as mash to 12 birds housed individually in laying cages from 7 to 20 weeks of age. The birds in different dietary groups had earlier received the respective quail starter±grower diets containing graded amounts of NKM as in the layer quail diets. Observations on egg production of quails, feed intake, blood biochemicals and pathomorphology of vital organs were made. Results on feed intake revealed insigni®cant differences amongst the groups and almost similar egg production and egg quality. However, quails fed the NKM diets tended to utilize feed less ef®ciently (P < 0:01) as compared to those fed the control diet. Except for signi®cantly high (P < 0:01) ALP activity in quails fed the NKM diet, no signi®cant (P < 0:05) differences in haematological (Hb, TEC, TLC), serum biochemicals (glucose, protein, cholesterol), and the AST, ALT activities were noticed. Regression analysis of data showed a decreasing trend of egg production with poor feed:egg and an increased TLC, protein and ALP as the dietary level of NKM was increased. Histopathology of vital organs of quails fed the NKM diets showed massive fatty in®ltration in liver and degenerative changes in the kidney tissues. From the results of the study it may be concluded that feeding NKM to Japanese quails from day 1 to 20 weeks of age supported normal feed intake, egg production and egg quality but, however, the ingredient on long-term feeding induced pathological changes in liver and kidney tissues. # 2000 Elsevier Science B.V. All rights reserved. Keywords: Quails; Neem kernel meal; Egg production; Haematology; Histopathology
*
Corresponding author. Tel.: 91-581-447223; fax: 91-581-447321. E-mail address: [email protected] (S.V.S. Verma). 0377-8401/00/$ ± see front matter # 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 7 7 - 8 4 0 1 ( 0 0 ) 0 0 1 8 4 - X
114
A.V. Elangovan et al. / Animal Feed Science and Technology 88 (2000) 113±120
1. Introduction The neem (Azadirachta indica) kernel meal (NKM) obtained on solvent extraction of neem seeds is available in sizable quantities of about 0.7 million MT per annum in India (Singh, 1993). However, despite of its high protein content of 300±400 g kgÿ1 the meal could not earn favour as a good livestock feed due to primarily to its pungent smell and bitter taste imparted by the presence of toxic triterpenoids; azadirone, nimbin, nimbidin, salanin, etc., instead utilized for fertilizer-cum-pesticide and also as a ¯y repellent. Several workers (Sadagopan et al., 1981; Verma et al., 1998; Gowda et al., 1998) tried to investigate the feeding value of NKM in the diet of laying hens but with variable success. Similar was the case with chicks (Sadagopan et al., 1982; Reddy and Rao, 1988; Chand, 1987; Jana, 1997). In a short-term study with laying WL hens, Gowda et al. (1998) recommended a safe level of inclusion of NKM 100 g kgÿ1 in diet. Thus the present study was undertaken with laying Japanese quails to examine the effect of inclusion of NKM in diet without adversely affecting their productive performance on long-term feeding. 2. Materials and methods 2.1. Experimental diet and birds One hundred and twenty Japanese quail chicks as hatched were randomly distributed into 12 lots of 10 each and housed in battery cages under similar managerial conditions. A maize soybean meal standard starter±grower mash (D1) with 230 g CP kgÿ1 and 11.27 MJ ME kgÿ1 was made. Three more diets (D2, D3 and D4) containing 50, 75 or 100 g NKM kgÿ1, respectively, for part of soybean meal and deoiled rice bran in diet D1 were made. One of the four diets was offered ad libitum to triplicate lots of chicks from day 1 to 6 weeks of age. At this age, 12 female quails from each of the four dietary groups were picked up and housed in individual laying cages. The respective grower diet were modi®ed in respect of CP and Ca contents to serve as layer diets without altering the NKM contents (Table 1). Each of the four test diets (D1, D2, D3 and D4) were offered ad libitum as mash to the corresponding quails from 7 to 20 weeks of age during which observations on net feed intake and daily egg production were recorded. The NKM and experimental diets were analysed for proximate composition, calcium and phosphorus as per standard procedures (AOAC, 1990). 2.2. Egg quality Ten eggs from each of the four dietary groups laid on ®rst two consecutive days during the 11th, 15th and 19th week of experimental feeding (i.e. 10 3 4 120) were utilized for quality testing in respect of shape index (100 times the ratio of width to length of an egg), albumen index (the ratio of average albumen height to the average of the width and length), yolk index (the ratio of yolk height to the average of the width), internal quality unit (IQU) as per Kondiah et al. (1981), shell weight and shell thickness.
A.V. Elangovan et al. / Animal Feed Science and Technology 88 (2000) 113±120
115
Table 1 Ingredient and nutrient composition of layer diets Dietary group (g NKM kgÿ1)
Attribute
D1 (0)
D2 (50)
D3 (75)
D4 (100)
720 220 60 ±
720 182 48 50
720 162.5 42.5 75
720 145 35 100
Nutrient composition (g or MJ kgÿ1), calculated Crude protein 186.1 Metabolisable energy 11.27 Lysine 9.80 Methionine 3.34
186.2 11.25 9.60 3.42
186.1 11.23 9.49 3.46
186.6 11.22 9.42 3.51
Nutrient composition (g or kgÿ1), determined Crude protein 186.9 Calcium 28.8 Phosphorus 9.33
182.5 28.2 9.35
183.1 29.5 9.41
184.2 30.1 9.12
ÿ1
Ingredient composition (g kg ) Basal mixturea Soybean meal (SBM) Deoiled rice bran (DORB) NKM
a Basal mixture consisted of g kgÿ1 diet: maize, 590; ®sh meal, 50; dicalcium phosphate, 14.3; ground lime stone, 30; oyster shell, 30; common salt, 3.0; trace mineral supplement (per kg diet provided: Mn, 55 mg; I, 12 mg; Fe, 156 mg; Zn, 30 mg; Cu, 4 mg; Co, 0.9 mg), 1.0; vitamin mixture (per kg diet provided: Vit. A, 8250 IU; Vit. D3, 1200 ICU, Vit. K, 1 mg; Vit. E, 40 IU; Vit. B1, 4 mg; Vit. B2, 4 mg; Vit. B12, 4.0 g; niacin, 60 mg; pantothenic acid, 10 mg; choline, 500 mg), 1.7%.
2.3. Organoleptic evaluation Ten eggs from each dietary group laid on the 3rd and 4th day of 11th, 15th and 19th week of feeding (i.e. 10 3 4 120) were evaluated organoleptically on an eight point hedonic scale in respect of colour, ¯avour, taste, smell, odour, texture and overall acceptability by a panel of 10 semitrained judges. For this purpose, eggs were boiled for 5 min and cooled under tap water. 2.4. Haematological and biochemical pro®le For the purpose of haemato-biochemical pro®le, blood samples from six quails per dietary group were drawn into a heparinized (20 IU/ml) glass vials at the end of 20 weeks of experimental feeding for the estimation of haemoglobin (Schalm et al., 1975), total erythrocyte (TEC) and total leucocyte counts (TLC) (Natt and Harrick, 1952). Concomitantly, the serum separated from blood drawn into another set of glass vials without any anticoagulants were stored at ÿ208C for subsequent use to estimate protein (Hiller and Slyke, 1927), cholesterol (Wybenga and Pileggi, 1970), ALT (Reitman and Frankel, 1957), AST (Reitman and Frankel, 1975) and ALP (Kind and King, 1954). The estimation of serum glucose (Cooper and McDaniell, 1970) was carried out immediately.
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2.5. Gross and histopathology Tissues from target organs (intestine, liver, spleen, heart, kidneys and ovaries) were collected from six birds per dietary group at the end of 20 weeks of experimental feeding were examined for gross and histopathological changes to see the effect of feeding NKM. The tissues were preserved in 10% formalin and processed as per standard techniques. 2.6. Statistical analysis The data in respect of various parameters were analysed statistically for treatment effects in a completely randomized design as per Snedecor and Cochran (1967). The data were further subjected to test the linearity among the graded levels of NKM feeding. 3. Results On chemical analysis, each kg of NKM DM was found to contain 382 g CP; 33 g EE; 132 g CF and 118 g total ash. An initial signi®cant (P < 0:01) drop in egg production due to feeding of NKM containing diets to quails improved gradually after 8th week of feeding (Table 2). The overall feed intake and egg production of quails in different groups Table 2 Laying performance of quails fed NKM diets and egg quality Attribute
Dietary group (g NKM kgÿ1) D1 (0)
D2 (50)
D3 (75)
D4 (100)
SEM
Production performance (7±20 weeks of age) Egg production, hen day (%) 7±8 weeks 82.0 7±20 weeks 88.9 Feed intake, 7±20 wk (g/b/d) 27.0 Egg weight (g) 11.6 Feed:egg 2.62 Live weight change 7±20 weeks (g/b) 26.2
62.5 85.3 28.0 11.5 2.86 31.0
55.9 85.8 26.3 10.7 2.91 25.6
57.5 83.8 27.3 11.1 3.00 23.6
6.49 1.72 0.68 0.32 0.062 4.16
Egg quality characteristics Shape index Shell thickness (mm) Shell weight (% of egg weight) Albumen index IQU Yolk index
77.9 0.193 10.5 0.129 62.3 0.478
78.0 0.203 10.5 0.131 62.7 0.467
77.6 0.193 10.4 0.130 63.1 0.471
77.7 0.200 10.4 0.133 63.7 0.471
0.98 0.0143 0.41 0.0063 1.21 0.0125
7.0 7.0 6.8 6.7 6.9 7.0
6.8 6.7 6.7 6.7 6.6 6.9
6.6 6.6 6.9 6.8 6.6 6.8
6.8 6.8 6.7 6.7 6.7 6.9
Organoleptic evaluation of boiled eggs Colour Flavour Taste Smell Texture Overall acceptability
0.33 0.41 0.38 0.29 0.35 0.28
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Table 3 Haemato-biochemical pro®le of quails fed NKM diets Attribute
Hb (g/dl) TEC (106 mmÿ3) TLC (103 mmÿ3) Glucose (mg/dl) Protein (g/dl) Cholesterol (mg/dl) ALT (IU/L) AST (IU/L) ALP (IU/L)
Dietary group (g NKM kgÿ1 diet) D1 (0)
D2 (50)
D3 (75)
D4 (100)
SEM
11.2 3.58 22.1 242 3.78 268 18.6 85.3 39.5
10.9 3.42 24.1 231 3.86 269 17.6 86.2 49.2
10.5 3.39 24.0 242 4.01 275 16.8 83.2 53.8
10.0 3.55 24.6 225 4.09 252 18.5 87.5 55.6
1.23 0.443 1.56 17.3 0.020 15.5 0.84 4.37 2.76
for the 20-week period did not differ signi®cantly (P > 0:05) due to dietary treatments. However, quails fed NKM containing diets required signi®cantly (P < 0:01) more feed for producing a unit egg mass. The pooled means at 11th, 15th and 19th week of experimental period for egg quality characteristics in terms of shape index, shell thickness, shell weight, albumen index, IQU and yolk index revealed no signi®cant (P > 0:05) difference assignable to dietary treatments (Table 2). Data on organoleptic evaluation of boiled eggs also did not show up any untoward characteristics in terms of colour, ¯avour, taste, smell, texture or the overall acceptability of the product that could be assignable to dietary treatments. The haematological and biochemical characteristics evaluated in terms of haemoglobin, TEC, TLC, glucose, protein and cholesterol revealed no signi®cant (P > 0:05) differences attributable to the feeding of NKM to quails (Table 3). Of the activity of various enzymes, the activity of ALP was found signi®cantly (P < 0:01) higher for quails fed the NKM diets than those of quails receiving the reference diet. The activities of ALT and AST, however, remained unaltered. Quails in all of the four treatment groups were in the state of positive weight gain and also no mortality assignable to the feeding of NKM were recorded during the experimental period. The linear regression equation of laying performance of quails and the haematobiochemical pro®le has been presented in Tables 4 and 5. The percent egg production of quails at 7±8 weeks and 7±20 weeks of age showed a signi®cant decrease (P < 0:065 and P < 0:057, respectively) with poorer feed:egg (P < 0:011) at higher levels of NKM in diet. Among the egg quality characteristics, the IQU showed a signi®cant increase (P < 0:036) as the level of NKM increased. As for haematobiochemicals, there was a decrease in haemoglobin (P < 0:042) and an increase in TLC (P < 0:058), protein (P < 0:035), ALP (P < 0:012) as the level of NKM was increased. Rest of all the parameters did not reveal any signi®cant linearity with graded levels of NKM in diet. The gross pathological examination of liver of quails fed diets D2±D4 containing NKM revealed slight enlargement with pale and greyish discolouration with petechieal haemorrhages, whereas the examination of kidney showed mild to moderate degree of
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Table 4 Linear regression equations of laying performance of quails fed NKM diets and egg quality r2
Significance (P)
Production performance (7±20 weeks of age) Egg production, hen day (%) 7±8 weeks Y 79:28 ÿ 0:26X 7±20 weeks Y 88:61 ÿ 0:047X Feed intake, 7±20 weeks (g/b/d) Y 27:21 ÿ 0:001X Egg weight (g) Y 11:62 ÿ 0:007X Feed:egg Y 2:636 0:0038X Live weight change 7±20 weeks (g/b) Y 28:19 ÿ 0:028X
0.87 0.89 0.005 0.52 0.98 0.15
0.065 0.057 0.931 0.276 0.011 0.614
Egg quality characteristics Shape index Shell thickness (mm) Shell weight (% of egg weight) Albumen index IQU Yolk index
Y Y Y Y Y Y
77:95 ÿ 0:0027X 0:1947 0:00004X 10:51 ÿ 0:0011X 0:129 0:0003X 62:19 0:0135X 0:476 ÿ 0:00007X
0.41 0.14 0.71 0.69 0.93 0.40
0.359 0.624 0.155 0.171 0.036 0.371
Organoleptic evaluation of boiled eggs Colour Flavour Taste Smell Texture Overall acceptability
Y Y Y Y Y Y
6:93 ÿ 0:0022X 6:91 ÿ 0:0024X 6:84 ÿ 0:0006X 6:68 0:0005X 6:86 ÿ 0:0026X 6:98 ÿ 0:0009X
0.45 0.41 0.049 0.22 0.66 0.31
0.330 0.358 0.779 0.532 0.188 0.447
Attribute
Linear equation
congestion. Microscopic examination showed a varied degree of degenerative changes in liver paranchyma with extensive fatty and mononuclear in®ltration even in those of quails fed the low level of 50 g kgÿ1 NKM in diet. Similarly, mild to moderate degree of degenerative changes were observed in the kidney tissues. The gross and microscopic examination of other vital organs revealed a normal picture of the tissues with no untoward changes assignable to NKM feeding.
Table 5 Linear regression equations of haemato-biochemical pro®le of quails fed NKM diets Attribute
Linear equation
r2
Significance (P)
Hb (g/dl) TEC (106 mmÿ3) TLC (103 mmÿ3) Glucose (mg/dl) Protein (g/dl) Cholesterol (mg/dl) ALT (IU/L) AST (IU/L) ALP (IU/L)
Y Y Y Y Y Y Y Y Y
0.92 0.10 0.89 0.39 0.93 0.21 0.09 0.05 0.98
0.042 0.678 0.058 0.376 0.035 0.543 0.706 0.779 0.012
11:31 ÿ 0:012X 3:52 ÿ 0:0007X 22:34 0:024X 241:9 ÿ 0:12X 3:76 0:003X 271:9 ÿ 0:105X 18:20 ÿ 0:0058X 85:02 0:009X 40:15 0:17X
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4. Discussion The initial egg production of quails fed the NKM containing diets appeared to be signi®cantly (P < 0:01) lower as compared to those fed the soybean meal containing reference diet and which may have been due to delayed sexual maturity. Such an effect was, however, not indicated by earlier workers (Verma et al., 1998; Gowda et al., 1998) in WL hens probably because the NKM was not introduced in diet from day one as has been done in the present study. However, the adverse effects of NKM feeding to quails were not re¯ected in the overall egg production which was found comparable among the different dietary groups as has been reported by earlier workers with WL hens (Sadagopan et al., 1981; Verma et al., 1998; Gowda et al., 1998). The overall feed intake among quails on different diets was also found comparable as has been experienced by earlier workers in WL hens fed NKM up to 100 g kgÿ1. But in the study of Gowda et al. (1998), they observed a lowered egg production and feed intake by WL hens fed NKM in diet at levels more than 100 g kgÿ1. However, in the present study, a decreasing trend was observed both at initial as well as overall egg production with graded levels of NKM in diet on regression analysis. In spite of similar egg production rate and feed intake of quails in different dietary groups, feed required for unit egg production was higher (P < 0:01) in those birds fed NKM diet than those on reference diet and such observations corroborate to the ®ndings of Sadagopan et al. (1981) in WL hens. On the contrary, earlier works of Verma et al. (1998) and Gowda et al. (1998) did not observe any signi®cant effect on feed:egg ratio in WL hens fed NKM up to 100 g kgÿ1 diet. The mean egg weight and body weight changes of quails in different dietary groups being similar and that all the birds showed positive growth. The egg quality characteristics as well as the organoleptic evaluation of boiled quail eggs were almost similar for all the dietary groups as also observed by earlier workers (Verma et al., 1998; Gowda et al., 1998) while employing WL hens for their study indicating that the bitter taste of triterpenoids were not transferred to the eggs. Feeding NKM to quails appeared to have not in¯uenced the haemato-biochemical pro®le except for a higher (P < 0:01) ALP activity in quails as compared to those fed the reference diet. Further, a increasing trend of TLC, protein and ALP on regression analysis provided a supportive evidence to the pathological alterations observed in some of the vital organs examined particularly the liver and kidney of the NKM fed quails. The absence of noticeable pathological lesions in WL hens fed 100 g NKM kgÿ1 diet for a 12 weeks period from 25 to 37 weeks of age (Gowda et al., 1998) could, therefore, be argued in terms of a relatively shorter feeding period. Moreover, the introduction of NKM in diet of quails right from day one of age may have accentuated the metabolic changes leading to signi®cant pathological lesions. Thus, it could be concluded that basing on the pathological changes observed in the vital organs, the overall use of NKM at any signi®cant level in the diet of quails may not be very bene®cial. References AOAC, 1990. Of®cial Methods of Analysis, 15th Edition. Association of Of®cial Analytical Chemists. Washington, DC.
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