Revista Científica UDO Agrícola Volumen 9.
Número 4. Año 2009. Páginas: 979-985
Growth
performance and haematology of Clarias
gariepinus (Burchell,
1822) fed varying inclusions of Leucaena leucocephala seed meal based-diets
Evaluación del crecimiento y
hematología de Clarias gariepinus (Burchell, 1822) alimentados con diferentes inclusuiones de dietas basadas en harinas de semillas de Leucaena leucocephala
Akeem Oladipupo SOTOLU 1 and Emmanuel Olujimi FATUROTI2
1Department of Forestry Wildlife and Fisheries Management, Faculty of
Agriculture, Nasarawa State University. Keffi, Lafia Campus, P. M. B. 135 Lafia, Nigeria and 2Department
of Wildlife and Fisheries Management, University of Ibadan, Ibadan, Nigeria
E-mails: sotoluola@yahoo.com Corresponding author
Received: 06/08/2009 |
First reviewing ending: 08/10/2009 |
First review received: 09/03/2009 |
Second reviewing ending: 09/13/2009 |
Second review received: 11/15/2009 |
Accepted: 12/27/2009 |
ABSTRACT
Utilization of plant protein sources in aquaculture have continued to
produce more promising results towards alleviating high cost of feeding. This study examined the utilization of Leucaena leucocephala
seed meal (LSM) for sustainable fish production. Six isonitrogenous;
40% crude protein diets were formulated where LSM replaced Soya bean meal (SBM)
at 0%, 20%, 40%, 60%, 80% and 100% inclusions. Catfish fingerlings (5.21±0.14g)
stocked at 25 fish/70 liters tanks were fed diets in triplicates three times
daily for 112 days. Solid wastes were siphoned everyday
before feeding while total water exchange with fresh clean water was done when
data on fish growth and haematology were collected.
Chemical analysis of diets and feeds were carried out before experiment and
that of fish alone was repeated after the experiment. Data from the completely
randomized experiment were subjected to ANOVA and correlation analysis and
L.S.D. was separated at 5% probability level. Mean weight gain (MWG) and
Specific growth rate (SGR) of fish fed 20% LSM were statistically different
(p<0.05) from those fish fed LSM at higher inclusion rates. Fish MWG, SGR,
PER and FCR significantly (p<0.05) correlated negatively with LSM inclusion
rates in fish diet r= -0.62,-0.57,-0.78 and -0.64 respectively. Fish carcass protein, packed cell volume and haemoglobin counts of fish were statistically the same for
0%, 20% and 40% LSM fed fish. In the present study processed leucaena seed meal can be considered as a good alternative
raw material in substitution to soya bean meal for Clarias gariepinus fingerlings’ diets at 20%
inclusion level.
Key words: Plant protein, leucaena seed meal, soya bean
meal, fish feeding, aquaculture
RESUMEN
La
utilización de fuentes de proteínas vegetales en la acuicultura ha continuado
para producir más resultados prometedores para aliviar el alto costo de la
alimentación. Este estudio examinó la utilización de harina de semillas de Leucaena (HSL) para la producción sustentable de peces. Se
formularon 6 dietas isonitrogénicas, dietas con 40%
de proteína cruda donde HSL reemplazó a la harina de soya (HS) a 0, 20, 40, 60,
80 y 100% de las inclusiones. Alevines de bagre (5,21 ± 0,14g) almacenados en
tanques de 25 peces/70 litros se alimentaron con dietas en triplicados, tres
veces diariamente durante 112 díoas. Los desechos
sólidos se sifonearon todos los días antes de la
alimentación mientras el cambio total de agua con agua fresca y limpia se
realizó cada dos semanas cuando se colectaron los datos del crecimiento de los
peces y laos caracteres hematológicos. Se llevaron a
cabo análisis químico de las dietas y los alimentos antes del experimneto y el análisis sólo de los peces se repitió
después del experimneto utilizando el método de la
AOAC (2000). Se sometieon los datos del experimneto completamente aleatorizado a ANAVA y análisis
de correlación y la prueba de MDS se usó para separar los tratamientos a un 5%
d eprobabilidad de avuerdo
a Steel et al. (1997). La ganancia
promedio de peso (GPP) y la tasa de crecimiento específico (TCE) de los peces
alimentados con 20% HSL fueron estadísticamente diferentes (p < 0,05) de
aquellos peces alimentados con HSL a tasas más altas de inclusuión.
GPP, TCE, PER y FCR significativamente (p < 0,05) se correlacionaron con las
tasas de inclusión de HSL en la dieta de los peces, r = -0,62, -0,57, -0,78 y
-0,64, respectivamente. Laz proteína en el canal de
los peces, Volumen de células empaquetadas y conteos de hemoglobina de los pecesfueron estadístimacamente
similares para peces alimentados con 0, 20 y 40% HSL LSM. La harina de semillas
de Leucaena ofrece buenos valores nutritivos en la
dieta de peces y su utilización fue mejor a 20% de inclusión para la producción
sustentable de peces.
Palabras clave: Proteína vegetal, harina de semillas de Leucaena, harina de soya, alimentación de peces,
acuicultura
INTRODUCTION
Lack
of readily available nutritive fish feed ingredients have continued to be a
major constraint to the survival of aquaculture in the competitive global food
production system (Ogunji et al., 2005; F.A.O
2006). Consequently, fish nutrition experts world over have considered the
recruitment of alternative protein feed ingredients necessary for inclusion in
fish diet. Several studies have shown that vegetable protein sources have high
potentials for supplying fish with required protein needed for their maximum
productivity (Hasting, 1976; Nwanna et al.,
2008). However, in the compounding of fish ration with plant protein sources,
cautions need to be exercised as to their inclusion levels in fish diets as
well as ensuring their proper processing for effective utilization (Pillay, 1990; Francis et
al., 2001). The need for such recommendations have been due to the presence
of certain limiting factors in those ingredients such as high crude fibre content (Nwanna et al.,
2008), antinutritional factors such as in Vigna subtarreana (Alegbeleye et al., 2001). Studies have shown that,
excessive consumption of plant protein sources by fish could cause slower
growth rates and poor performance which may result in mortalities if condition
persists (Francis et al., 2001; Cho, et al., 1974).
Leucaena leucocephala demonstrated good
potential to serve as a useful plant protein source in fish ration and in the
livestock industry generally (Jones, 1979; Ter Muelen et al., 1981; D’Mello
and Acamovic, 1989). However, it has been established
that leucaena contain mimosine-a
non-protein amino acid capable of inhibiting protein biosynthesis in animal
causing growth retardation if consumed intensively (Ter
Muelen et al., 1981; D’Mello
and Acamovic, 1989).
Cruz and Laudencia (1977) found out that 33 to
100% leucaena leaf meal as integral part of
supplemental feed enhanced the growth of Oreochromis
niloticus fingerlings in Lake Laguna. D’Mello and Acamovic (1989) on
the other hand, stressed its potential as a good feed ingredient in the culture
of mollies and topminnows (Poecilia spp) and freshwater prawn (Macrobrachium
rosenbergii). A preliminary study by Sotolu and Faturoti (2008)
revealed that catfish was able to digest leucaena
seed meal (LSM) processed by soaking in water than those LSMs processed by
other methods. Leucaena seed meal was therefore used
to replace the much expensive and scarce soybean meal (SBM) in catfish diets in
this study. It involved the evaluation of the effects of leucaena
seed meal on growth performance of Clarias gariepinus, protein utilization and carcass
composition. This study examined the utilization of L. leucocephala
seed meal (LSM) for sustainable fish production
MATERIALS AND
METHODS
Preparation of leucaena seed
meal and experimental set-up
Whole
pure leucaena seeds were soaked in cold water at 1
kg/5 litres of water for 72 hours as described by Padmavathy and Shobha (1987).
Seeds were later thoroughly washed in fresh cold water after removal thorough
sieve. Seeds were spread in thin layer on a slab for quick and homogenous solar
drying for two days. Samples of soaked in cold water leucaena
seeds (Processed leucaena seed meal-PLSM) were
chemically analyzed according to A.O.A.C (2000). Parameters of importance
include crude protein, crude fibre, fat, ash,
moisture and calorific value. Six isonitrogenous
diets were prepared (40% C.P) where the processed leucaena
seed meal (LSM) replaced soyabean meal (SBM) at 0%,
20%, 40%, 60%,80% and 100%.
All
ingredients used in the formulation were ensured constant except SBM and LSM
that were made to substitute for one another on percentage basis shown. Slight
variations however occurred in the crude protein content of the formulated
diets on chemically analysis and this may be due to differences in their
compositions. Values of their crude protein ranged
between 39.86% and 40.29%.
Chemical
analysis of experimental feeds and fish were done before feeding trials using
the method of A.O.A.C (2000) while that of the experimental fish alone was
repeated at the end of the experiment. Water quality parameters (pH, Dissolved
oxygen, ammonia) were kept favorable for fish by constant renewal of water in
fish tanks at a flow rate of 2.5l/mins till the end
of the experiment. Catfish fingerlings (5.21+0.14g) were stocked in a
circular tank of 70 liter capacity at 25 fish/tank in
triplicates. Feeding commenced three days after acclimation of fish
sourced from the University of Ibadan fish hatchery. Fish were fed to satiation
between the hours of 09:00h-09:30h, 14:00h-14:30h and 19:00h-19:30h daily for
112 days. Solid wastes were siphoned out of tanks everyday
before feeding and total cleaning of fish tanks and replacement with clean
fresh water was done every fortnight.
Determination of feed utilization and data analysis
Data
were collected fortnightly on fish growth performance and nutrient utilization
by determining mean weight gain (MWG), Gross energy was calculated according to
Jobling (1983) with multiplier factors of
carbohydrate, 4.1 kcal/g, protein, 5.4kcal/g and lipids, 9.5kcal/g.
Where,
a = Initial mean
weight of fish
b= final mean
weight of fish
h= experimental
period in days
Fish
were tranquilized with 150mg/L of Tricane methane Sulphonate for blood collection according to Schalm et al. (1975). Initial fish blood samples were
collected before feeding trial that is 0th day with the aid of needle and
syringe into bottles with EDTA as anticoagulant. Subsequent blood samples were
collected on 42nd, 56th, 70th, 84th,
98th and 112th day as the final collection. Blood samples
collected were immediately taken to the laboratory for haematological
analysis on each occasion as described by Schalm et
al. (1975).
Data
collected were subjected to ANOVA and correlation analysis using the SPSS
package version 10 and significant mean differences were separated at 0.05
probability level according to Steel et al., (1997).
RESULTS
Proximate
composition of processed and unprocessed leucaena
seeds is given in Table 1. Processing increased protein content from 22.7 to
36% while crude fiber decreased from 11.4 to 7.1 in the unprocessed and
processed leucaena respectively. Diet formulation
gross and proximate compositions of the six diets formulated for the feeding
trial are presented in Table 2. Values for crude protein content ranged between
39.86% to 40.20% while crude lipid ranged from 9.87 to 10.36%..
Growth performance feed and protein utilization is given in Table 3. Highest
mean weight gain (MWG) was recorded for fish fed 0% and 20% LSM based-diets
7.9g and 7.7g respectively which were significantly (p<0.05) higher than the respective values
of the other LSM inclusions while 40% and 60% LSM based diet were only
marginally different (P<0.05). MWG of fish continue to
decrease with increase in the LSM inclusion rate as 80% LSM and 100% produce
the least set of values (4.67g and 4.07g) respectively, which were also not
significantly different. Specific growth rate (SGR) was significantly higher
(P<0.05) in fish fed (20%LSM) than all other treatment including control
(0%LSM) which was almost the same with those of 40% and 60%LSM. Fish fed 80%
and 100% LSM were only marginally different 0.25%/day and 0.23%/day
respectively. Fish growth exhibited significant inverse correlation with
inclusion rate of LSM in the diet formulated. MWG and SGR had a -0.62 and -0.57
correlation coefficients (r) respectively while PER and FCR had -0.78 and
-0.64, respectively (data not shown).
Table 1. Proximate and mineral composition of
unprocessed (ULSM) and processed leucaena seed meal
(PLSM). |
|||||||||||
LSM |
Crude protein
(%) |
Crude fibre (%) |
Fat (%) |
Ash (%) |
Moisture (%) |
Energy kcal/ kg |
Na (%) |
K (%) |
P (%) |
Ca (%) |
Mg (%) |
ULSM |
22.75 |
11.38 |
6.12 |
5.98 |
15.14 |
2833.50 |
0.01 |
1.62 |
0.46 |
0.41 |
0.22 |
PLSM |
36.01 |
7.11 |
5.18 |
3.74 |
12.56 |
2899.76 |
0.03 |
1.68 |
0.68 |
0.42 |
0.29 |
Table 2. Diet formulation and proximate
composition of LSM based-diets in replacement of SBM at 0% - 100%). |
||||||
Gross composition ingredients
(g/100g/DM) |
Diet 1 |
Diet 2 |
Diet 3 |
Diet 4 |
Diet 5 |
Diet 6 |
0% |
20% |
40% |
60% |
80% |
100% |
|
SBM |
23.39 |
19.51 |
14.64 |
9.75 |
4.88 |
- |
LSM |
- |
4.88 |
9.75 |
14.64 |
19.51 |
24.39 |
Fishmeal |
18.29 |
18.29 |
18.29 |
18.29 |
18.29 |
18.29 |
Groundnut
cake |
26.59 |
26.59 |
26.59 |
26.59 |
26.59 |
26.59 |
Yellow maize |
27.38 |
27.38 |
27.38 |
27.38 |
27.38 |
27.38 |
Bone meal |
1.50 |
1.50 |
1.50 |
1.50 |
1.50 |
1.50 |
Oyster shell |
1.35 |
0.25 |
0.25 |
0.25 |
0.25 |
0.25 |
Palm oil |
0.75 |
0.75 |
0.75 |
0.75 |
0.75 |
0.75 |
Mineral /vit. supplements* |
0.60 |
0.60 |
0.60 |
0.60 |
0.60 |
0.60 |
Chemical
composition (%) on a dry matter basis |
|
|||||
Crude protein |
40.14 |
40.02 |
39.97 |
39.86 |
40.18 |
40.20 |
Lipid |
9.93 |
10.27 |
9.98 |
10.01 |
9.87 |
10.36 |
Crude fibre |
2.41 |
2.39 |
2.56 |
2.58 |
2.57 |
2.81 |
Ash |
7.03 |
8.21 |
8.33 |
9.62 |
9.19 |
8.85 |
NFE |
36.32 |
35.74 |
35.58 |
38.01 |
36.33 |
38.25 |
Moisture |
4.92 |
5.66 |
5.62 |
5.17 |
4.84 |
5.03 |
Gross energy (kcal/kg) |
2890.41 |
2954.66 |
3000.42 |
2889.85 |
3086.00 |
3031.29 |
Biomix fish vitamin/mineral
providing per kg of diet at 5kg per tonne
inclusion: 20,000 iu, vitamin A, 200 i.u, α-tocopherol acetate
400mg, Ascorbic acid 100mg, Vit. D3, 200 mg Vit E, 8 mg Vit k3, 20mg Vit B1, 30 mg Vit B2, 12 mg Vit B6, 50 mg Pantothenic acid, 0.8 mg Biotin, 150 mg
Niacin, 0.05mg Vit B12, 4.0mg Cobalt, 40 mg Iron,
5.0 mg Iodine, 30 mg Manganese, 4 mg Copper, 40 mg Zinc, 0.2 mg Selenium, 100
mg Lysine, 100 mg Methionine, 100 mg Anti-oxidant. |
Table 3. Growth and nutrient utilization of Clarias gariepinus
fed different inclusions of LSM based-diets. |
|||||||
|
Treatments-LSM inclusion rates
(%) |
||||||
Parameters |
0 |
20 |
40 |
60 |
80 |
100 |
SEM |
Initial
mean weight (g) |
5.21+0.14 |
5.29+0.14 |
5.09+0.14 |
5.22+0.14 |
5.20+0.14 |
5.23+0.14 |
- |
Final mean weight (g) |
13.16+0.02 |
12.98+0.17 |
11.94+0.11 |
10.97+0.08 |
9.87+0.11 |
9.30+0.10 |
- |
MWG (g) |
7.95a |
7.69a |
6.85b |
5.75b |
4.67c |
4.07c |
0.65 |
Feed intake (g)/fish |
56.78a |
57.51a |
54.40c |
49.98d |
54.54bc |
55.18b |
1.08 |
PWG (%) |
60.41a |
59.24a |
57.37a |
52.42b |
47.32c |
43.76c |
2.77 |
SGR (%/day) |
0.36b |
0.46a |
0.33b |
0.39b |
0.25c |
0.23c |
0.04 |
PER |
0.35a |
0.33a |
0.32a |
0.29a |
0.21b |
0.18b |
0.03 |
FCR |
0.71c |
0.75c |
0.79bc |
0.90b |
1.17a |
1.36a |
0.11 |
Nm
(x10) |
56.48a |
56.17a |
52.66b |
49.77b |
46.33c |
44.67c |
2.02 |
NPU |
88.10a |
87.87a |
82.46b |
79.19b |
72.18c |
69.56c |
3.20 |
Survival
rate (%) |
92 |
92 |
88 |
92 |
76 |
80 |
- |
Means
with the same superscript in the same row are not significantly different
(p<0.05) |
Results
also showed that values of PER were only marginally different in fish fed 0% to
60%LSM (0.35 to 0.29) while values of FCR were only marginally different in
fish fed 0% to 40%LSM (0.71 to 0.79). In both parameters examined further increase of LSM in
the diet (60 and 100%) resulted in significant worsening. Protein utilization
expressed by Nm and NPU in fish decreased significantly from 40% LSM inclusion
level. Values of 0% and 20% LSM were statistically the same and similarly
values of 40% and 60% LSM fed fish, while values of 80% and 100% LSM had the
lowest set of values. Survival rate was jointly highest in 0%, 20% and 60% LSM
(92%) while 80% LSM had the least survival rate (76%).
The
effects on the physiological changes in fish fed graded levels of LSM based
diets for 112 day is presented in Table 4. Fish carcass protein increased
throughout in all diets. Initial carcass protein value was 62.08% while 0% LSM
produced 68.47% C.P and 100% LSM produced 66.48% C.P respectively. Fish carcass
lipid seems to increase in all LSMs with exception in 20% LSM where initial
value was reduced from 4.27% to 4.12%.
Table 4. Carcass and haematology
composition of Clarias gariepinus
fed LSM based-diets for 112 days. |
||||||||
Parameters (%) |
Initial |
Final values at Different LSM inclusion
rates, % |
||||||
|
Value |
0 |
20 |
40 |
60 |
80 |
100 |
SE mean |
Crude protein |
62.08 |
68.47 |
68.13 |
66.81 |
67.91 |
66.74 |
66.48 |
1.74 |
Lipid |
4.27 |
5.93 |
4.12 |
5.98 |
4.53 |
4.96 |
5.58 |
0.19 |
Ash |
11.60 |
11.14 |
10.83 |
10.94 |
11.02 |
10.86 |
10.80 |
0.50 |
Crude fibre |
ND |
ND |
ND |
ND |
ND |
ND |
ND |
- |
NFE |
5.23 |
2.47 |
3.02 |
3.01 |
2.78 |
2.82 |
3.94 |
0.29 |
Haematocrits |
|
|
|
|
|
|
|
|
PCV (%) |
28.21a |
28.22a |
28.20a |
29.16b |
28.15b |
28.02c |
27.99c |
0.06 |
Hb (gm/100ml) |
8.39a |
8.43a |
8.41a |
8.37a |
8.22b |
8.20b |
8.17b |
0.05 |
Rbc (x106/ml) |
2.36b |
2.49a |
2.44a |
2.38b |
2.34b |
2.31b |
2.24c |
0.05 |
Wbc (x103/ml) |
16.17a |
16.13a |
16.16a |
16.20a |
16.29b |
16.39c |
16.21a |
0.04 |
MCV (fl) |
119.53b |
113.44c |
114.23c |
115.98c |
121.20b |
127.29a |
130.37a |
2.46 |
ND – Not Detected. Means with the same superscript
in the same row are not significantly different (p<0.05) |
Fish PCV increased from initial 28.21% to
28.22% in 20% LSM and subsequently decreased marginally to 28.16% in 40% LSM.
The least PCV value was recorded for 100% LSM (27.99%) which was not
significantly different from 80% LSM (28.02%). Results of Hb
also increased from its initial value but up to value of 40% LSM where
differences were only marginal (P<0.05). Rbc increased in value from initial 2.36 (x106/ml)
to 2.49 (x106/ml) in 0% LSM. Rbc
was not significantly different in values for initial composition, 40%, 60% and
80% LSM while 100 LSM produced significant least value. Wbc did not vary significantly from initial value to
20% LSM inclusion but there was significant variation (P<0.05) between 40%,
60% and 80% LSM while 100% LSM was only marginally different from 40% LSM. The
highest value for MCV was found in fish fed 100% LSM while the least was
recorded for fish fed 0% LSM with significant variation (P<0.05) while
initial value 119.53(fl) was only marginally
different from the value of 60% LSM (121.20 fl).
DISCUSSION
Growth
and nutrient utilization by fish decreased as level of LSM inclusion increases
in the diets. This observed pattern could probably be a result of persistent
consumption of leucaena meals which could retard
animal growth rate as reported by Jones (1979) and further buttressed by Tangendijaja et al., (1990) who recorded progressive
depressed growth rate in rabbit fed increasing graded levels of leucaena leaf meal based-diet. Similarly the results on PWG
and SGR could also be due to differences in the LSM inclusion which decreased
at increasing level of LSM in the diets. The consumption of antinutrient
factors contained in the LSM based diets (mimosine
like) are probably responsible for retarded growth response of the fish.
Protein efficiency ratio (PER) was highest in fish fed 0% LSM but did not
differ statistically (p<0.05) from values of 20%, 40% and 60% LSM
inclusions. These results seem to have direct link with feed intake. The importance
of feed intake by fish as a determinant of fish performance has been strongly
emphasized (Preston and Leng, 1987; Faturoti, 1989; Pillay 1990)
while other studies (Anderson et al., 1984; Keembiyehetty
and De-Silva, 1993) pointed out the possibility of protein sparing effects by
other nutrients in a feed, that is as more energy was supplied for metabolism
through other nutrients, more protein is available for fish growth and tissue
development.
All
diets produced higher values of fish carcass protein and lipid than initial
values, yet there existed marginal difference among them indicating different
utilization levels of the diets. These relatively high values of crude protein
could be viewed alongside the work of (Alegbeleye et
al., 2001) who reported that effective utilization of bambara groundnut at varying rates was responsible
for variations in Heteroclaris carcass protein
and lipid. The non-detection of crude fiber in the fish carcass composition was
the same in all treatments and this had been said to be associated with
effective utilization of diets according to (Sotolu,
2008). Observed differences in the hematology of fish especially at significant
level (P<0.05) between 20% LSM fed fish and those of higher LSM inclusions
could be as a result of the residual effect of mimosine
present in the seeds after processing. This was similar to the reports of Adeyemo (2005) and Osuigwe et
al., (2005) that ascertained the reduction in values of haematological
parameters such as PCV, Hb and Rbc were due to the presence of toxic substances in
the diet of fish. It could therefore be inferred from this study that fish with
lower LSM inclusion levels were of better health status than those of higher
LSM inclusions based on the earlier submissions of Svobodova
et al., (1991), Alegbeleye (2005) and Ochang et al.,
(2007).
CONCLUSION
The
utilization of LSM by fish when processed by soaking in water was better at 20%
inclusion level than at higher inclusion rates. Since weight gain of fish is
what would translate into income for the fish farmer at the end of the
production cycle, 20% inclusion rate of LSM in catfish diet would produce
better and profitable result at present. However, there is still need for
further studies towards increasing the utilization level of LSM to 40%
inclusion as most utilization and haematological
parameters assessed in this study were marginally different from those of 20%
inclusion. Cost of fish production is expected to further reduce if more soya
bean meal could be replaced by leucaena seed meal.
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