Vol.1, No.2, 51-55 (2010)
doi:10.4236/as.2010.12007
Copyright © 2010 SciRes. Openly accessi ble at http://www.scirp.org/journal/AS/
Agricultural Sciences
Effects of treatment methods on the nutritional value of
cotton seed cake for laying hens
David Friday Apata
Department of Animal Production, University of Ilorin, Ilorin, Nigeria;
Received 17 June 2010; revised 8 July 2010; accepted 21 July 2010.
ABSTRACT
The effects of treatment methods on the nutri-
tive value of cotton seed cake (CSC) for laying
hens were examined. Olympia laying hens (n =
96) were assigned randomly to a 2 × 3 factorial
combination of fermented or unfermented CSC
supplemented with enzyme, Vitamin E and fer-
rous sulphate over a 4-week feeding period.
CSC was incorporated at 15% in the diets. Re-
sult s sho wed no significant differenc es (P > 0.05)
in feed intake, body weight gain and egg weight
due to treatments. However, hen-day egg pro-
duction was significantly reduced (P < 0.05)
among dietary treatments compared with the
control. Layers fed on unfermented CSC + Vi-
tamin E and unfermented CSC + FeSO4 had the
lowest (P < 0.05) values for packed cell volume
and haemoglobin. Treatment differences in red
blood cell and white blood cell were not sig-
nificant (P > 0.05). Serum concentration of total
protein among diet ary treatmen t s was lower (P <
0.05) than the control, while albumin and cho-
lesterol values were similar (P > 0.05) for all
treatments. Generally, activities of alanine and
aspartate aminotransferases increased (P < 0.05)
among CSC dietary treatments compared with
the control diet. The interaction between CSC
form and additives had no significant effect on
any of the parameters. The results of this study
indicate that the treatment methods employed
for CSC could not significantly improve its nu-
tritive value for egg production at 15% level in
the diet of laying hen.
Keywords: Cotton Seed Cake; Haematology;
Serum Constituents; Laying Hen
1. INTRODUCTION
One major limiting factor in the expansion of egg pro-
duction enterprise is the high cost of feedstuffs such as
soybean meal, groundnut cake and fish meal. This has
made egg production expensive, thereby worsening the
intake of protein sources by the populace of developing
countries. One approach to reduce pressure on conven-
tional protein ingredients is greater utilization of alterna-
tive sources of dietary protein alongside other strategies.
Cotton seed cake (CSC) is a by-product of the cotton
processing industry. The cake is rich in protein (42%)
but it contains gossypol which has been recognized since
the turn of the century to be toxic to animals [1,2]. In
monogastric animals, gossypol interfered with protein
digestion, bind lysine (making it unavailable) and re-
duced growth rate and productivity [3,4]. There is an-
other problem as the cake contains a high level of fibre,
perhaps 23% because undelinted seeds are used in the
processing [5]. This high fibre content has limited the
extent to which it can be used in the diets of poultry
which lack the appropriate enzymes capable of degrad-
ing fibre [6,7].
Currently, CSC is used by feed millers in Nigeria at
7.5% inclusion level with ferrous sulphate supplement to
inactivate gossypol in balanced layers feed. This chemi-
cal is usually not available to local farmers at farm sites.
It becomes imperative therefore that simple treatment
method be found that will overcome the gossypol and
degrade the high fibre in CSC so that its nutritive value
could be enhanced. Vitamin E is an antioxidant that pre-
vents the damage of free radicals at the cellular level [8].
This study was designed to investigate the effective-
ness of enzyme, vitamin E and ferrous sulphate treat-
ments in improving the nutritive value of CSC for laying
hens.
2. MATERIALS AND METHODS
Cotton seed cake was obtained from a feed miller in
Ilorin. Samples (20 kg) of the cake were placed in a dou-
ble layer polythene bag, tied up and placed in an empty
plastic drum, sealed and fermented for a period of 7 days.
Thereafter, the fermented cake was removed and sun-

D. F. Apata / Agricultural Sciences 1 (2010) 51-55
Copyright © 2010 SciRes. Openly accessible at http://www.scirp.org/journal/AS/
52
dried for 3 days. The fermented and a batch of the un-
fermented cake (20 kg) were used in preparation of the
diets described below.
Seven isonitrogenous (18% crude protein) and isoalo-
ric (2623 kcal/kg ME) experimental diets (Table 1) were
formulated by incorporating fermented or unfermented
CSC at 15% and supplemented with enzyme, vitamin E
and Ferrous Sulphate (FeSO4) at 0.01, 0.03 and 0.20%,
respectively in a 2 × 3 factorial design. Maize soybean
meal-based diet was used as control. The enzyme prepa-
ration (Allzyme) derived from Aspergillu s niger contains
amylase, beta-glucanase, cellulase, pectinase, phytase,
protease and xylanase activities. The level of enzyme
supplementation was chosen according to manufacturer’s
recommendation.
Ninety six hens of the Olympia strain that had been in
lay for 10 weeks were purchased from a commercial
poultry farm in Ilorin. The birds were individually caged
in a poultry pen which was illuminated at night and were
randomly assigned to the dietary treatments. The trial
lasted for 4 weeks. Feed intake was measured daily and
body weight weekly. Records of egg production per cage
swere kept daily throughout the experimental period. All
eggs laid each day were weighed individually.
Table 1. Composition of the experimental diets (%).
Unfermented Cotton Seed Cake Fermented Cotton Seed Cake Control
Ingredient 1 2 3 4 5 6 7
Maize 50.00 50.00 50.00 50.00 50.00 50.00 50.00
Wheat bran 12.64 12.62 12.45 13.14 13.12 12.95 15.85
Soybean meal 12.50 12.50 12.50 12.50 12.50 12.50 27.50
Cotton seed cake 15.00 15.00 15.00 15.00 15.00 15.00 0.00
Fish meal 1.00 1.00 1.00 1.00 1.00 1.00 1.00
Blood meal 3.50 3.50 3.50 3.00 3.00 3.00 0.30
Oyster shell 1.50 1.50 1.50 1.50 1.50 1.50 1.50
Bone meal 3.00 3.00 3.00 3.00 3.00 3.00 3.00
aPremix 0.25 0.25 0.25 0.25 0.25 0.25 0.25
DL-Methionine 0.15 0.15 0.15 0.15 0.15 0.15 0.15
Lysine 0.15 0.15 0.15 0.15 0.15 0.15 0.15
Salt (Nacl) 0.30 0.30 0.30 0.30 0.30 0.30 0.30
Enzyme 0.01 - - 0.01 - - -
Vitamin E - 0.03 - - 0.03 - -
FeSO4 - - 0.20 - - 0.20 -
bChemical Composition (%)
Crude protein 18.08 18.10 18.30 18.07 18.05 18.20 18.00
Crude fibre 5.36 5.31 5.32 5.34 5.30 5.34 4.71
Ether extract 4.01 4.01 4.01 3.92 3.92 3.92 3.89
ME (Kcal/kg) 2623 2623 2623 2625 2625 2625 2624
Lysine 1.17 1.17 1.17 1.19 1.19 1.19 1.22
Methionine 0.52 0.52 0.52 0.53 0.53 0.53 0.49
aProvided 1.5 mg retinol, 25 mg cholecalciferol, 16 mg α-tocopherol, 1mg menadione, 0.8 mg thiamine, 2.4 mg riboflavin, 14 mg nicotinic acid, 4
mg calcium D-pantothenate, 1.4 mg pyridoxine, 10 mg cyanocobalamin, 0.4 mg folic acid, 0.02 mg biotin, 120 mg choline chloride, 0.62 mg zinc
bacitracin, 36 mg avaten, 40 mg Mn, 18 mg Zn, 0.8 mg Cu, 0.09 mg Co, 20 mg Fe and 0.04 mg Se kg-1 diet; bDetermined values except for me-
tabolisable energy (ME), lysine and methionine which were calculated from the published (NRC, 1995) compositions of the ingredients used.

D. F. Apata / Agricultural Sciences 1 (2010) 51-55
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53
At the end of the experimental period, four hens were
randomly selected from each dietary treatment, fasted
overnight and blood samples collected from the wing
vein. Blood was collected into bottles containing EDTA
anticoagulant for haematology and clean dry bottles for
serum constituent analysis. Serum samples were sepa-
rated by centrifugation at 1600 × g for 15 min. and
stored at –20℃. The whole blood was analysed for
packed cell volume (PCV), haemoglobin (Hb), red blood
cell count (RBC) and white blood cell count (WBC)
using standard haematological techniques [10]. Serum
total protein, albumin, cholesterol, alanine amino trans-
ferase (ALT. EC 2.6.1.1) and aspartate amino transferase
(AST, EC 2.6.1.1) activities were measured using com-
mercial clinical investigation kits (Wako, Osaka, Japan).
3. STATISTICAL ANALYSIS
Data collected were subjected to a two-way factorial
analysis and Duncan’s multiple range test to determine
the significant difference at the 0.05 level [11].
4. RESULTS AND DIS CUSSION
Laying performance of hens was affected by dietary
CSC treatments at the end of the feeding trial (Table 2).
There were no significant differences (P > 0.05) in feed
intake, weight gain and egg weight due to treatments.
However, hen-day egg production was significantly re-
duced (P < 0.05) among dietary treatments compared
with the control. The highest egg production (56.7%) in
CSC diets achieved by hens fed fermented CSC + en-
zyme was lower (P < 0.05) than the control (67.00%).
Layers feed consumption and body weight gain were
unaffected by treatment methods, probably because the
fermentation process and the additives used in the diets
were unable to cause appreciable reduction in the gos-
sypol level. The feed consumed by the layers appeared
to be principally used for body maintenance. This view
is similar to that reported by Olorede and Longe [12]
that hens can use feed or metabolizable energy intake to
fulfill maintenance requirement, thereby leaving little for
egg production. In general, the decrease in egg produc-
tion observed with dietary treatments of CSC might be
the consequence of residual effect of gossypol that binds
with the cake’s protein thereby making it unavailable to
the hens for egg production. The marginal improvement
found with the fermented cake + enzyme may be attrib-
uted to the combination of fermentation process and en-
zyme degradation of the fibre that made nutrients avail-
able to the birds. This is in agreement with Babalola et al.
[7] who reported improved nutrient availability to pullet
chicks fed β-xylanase supplemented castor seed meal
diet.
The haematological values are presented in Table 3.
For all treatments, PCV and Hb were lower than the
control with the exception of fermented CSC + enzyme
which had similar PCV value with the control. No sig-
nificant differences (P > 0.05) were observed for RBC
and WBC. The decreases in PCV and Hb values in hens
fed diets containing unfermented CSC or fermented CSC
+ Vitamin E, or ferrous sulphate, suggest inadequate
nutrient utilisation in the hens. This may be partly due to
the residual effect of gossypol on blood variables. A re-
duction in haematological parameter has been reported
by Apata [13] and Rinchard et al. [5] in chicks and fish
fed diets containing high levels of legume seed meal and
cotton seed meal, respectively. However, Mitruka and
Table 2. Performance of laying hens fed diets containing different treatments of unfermented or fermented cotton seed cake (CSC).
Dietary treatments Feed intake
(g per hen day-1)
Weight gain
(g per hen day-1)
Hen-day Egg
production (%) Egg weight (g)
Unfermented CSC + enzyme 83.0a 3.4a 45.7b 54.6b
Unfermented CSC + Vit. E 86.7a 3.5a 44.3b 54.4b
Unfermented CSC + FeSO4 81.0a 2.9a 43.5b 53.2b
Fermented CSC + enzyme 85.8a 3.9a 56.7ab 56.7b
Fermented CSC + Vit. E 82.5a 3.1a 53.1b 56.4b
Fermented CSC + FeSO4 78.6a 3.4a 52.8b 55.0b
Control 106.5b 5.6b 67.0a 60.1a
F X A NS NS NS NS
SEM 7.2 1.1 6.4 3.8
Values in a column with the same letter do not differ significantly at P < 0.05. NS = not significant; SEM = standard error of means; F = test in-
gredient form; A = additives

D. F. Apata / Agricultural Sciences 1 (2010) 51-55
Copyright © 2010 SciRes. Openly accessible at http://www.scirp.org/journal/AS/
54
Tab le 3. Haematological values of laying hens fed diets containing different treatments of unfermented or fermented cotton seed
cake (CSC).
Dietary treatments PVC (%) Hb (g/dL) RBC (× 1012/L) WBC (× 109/L)
Unfermented CSC + enzyme 24.0ab 8.1ab 4.37 9.67
Unfermented CSC + Vit. E 18.0b 4.3a 3.50 11.30
Unfermented CSC + FeSO4 18.5b 5.1a 4.09 10.85
Fermented CSC + enzyme 27.0a 7.8ab 4.00 12.20
Fermented CSC + Vit. E 20.0b 6.0ab 4.03 9.97
Fermented CSC + FeSO4 22.3b 7.1ab 4.10 9.60
Control 30a 9.1b 4.90 9.10
F x A Ns Ns Ns Ns
SEM 1.6 0.41 0.19 1.02
Values in a column with the same letter do not differ significantly at p < 0.05; PCV = packed cell volume; Hb = haemoglobin; RBC = red blood
cell; WBC = white blood cell; Ns = not significant; F = tests ingredient form; A = additives; SEM = standard error of means
Table 4 . Serum biochemical constituents of laying hens fed diets containing different treatments of unfermented or fermented
cotton seed cake (CSC).
Dietary treatments Total protein
(mmol/L)
Albumin
(mmol/L)
Cholesterol
(mmol/L) ALT (IUL-1) AST (IUL-1)
Unfermented CSC + enzyme 23b 5.3 1.8 8.67b 118b
Unfermented CSC + Vit. E 26b 6.0 2.5 12.00b 116b
Unfermented CSC + FeSO4 22b 5.5 1.5 20.00c 216d
Fermented CSC + enzyme 24b 5.0 1.5 19.33c 163c
Fermented CSC + Vit. E 27b 5.3 1.5 19.33c 163c
Fermented CSC + FeSO4 26b 5.3 2.1 13.67b 134c
Control 31a 4.0 1.1 4.00a 90a
F x A Ns Ns Ns Ns Ns
SEM 1.08 0.67 0.90 1.06 13.86
Values in a column with the same letter do not differ significantly at p < 0.05; AST = aspartate aminotransferase activity; ALT = alanine ami-
notransferase activity; Ns = not significant; F = tests ingredient form; A = additives; SEM = standard error of means
Rawnsley [14] reported 7.00-18.6 g/dl as normal values
of Hb in chickens. Thus, a range of 4.3-5.1 g/dl obtained
in this study for unfermented CSC + Vitamin E, or FeSO4
fell short of the range consistent with good layers health.
Of the serum biochemical constituents, total protein
among dietary treatments was lower (P < 0.05) than the
control, whereas the ALT and AST were higher (P < 0.05)
than the control (Table 4), with no significant differ-
ences (P > 0.05) observed in albumin and cholesterol
values for all treatments. Decrease in serum total protein
concentration irrespective of the treatment method cor-
roborates the inadequacy of these simple technologies in
improving the nutritional quality of this feedstuff espe-
cially at the level of inclusion. The increase in serum
ALT and AST activities indicate release of the ami-
notransferases from cytoplasm to blood stream which is
probably due to liver and/or other tissues damage. A
similar observation has been reported by Muhammad
and Oloyede [15] for chicks fed femented Temialia cat-
tapa seed meal-based diet.
In conclusion, the results of this study show that the
different treatment methods employed for CSC could not
significantly improve its nutritive value for egg produc-
tion at 15% level in the diet of laying hen.

D. F. Apata / Agricultural Sciences 1 (2010) 51-55
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55
5. ACKNOWLEDGEMENTS
This study was partly funded by the senate research grant of the Uni-
versity of Ilorin. The author is grateful for this financial assistance.
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