East African Agricultural and Forestry Journal

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Effect Of Replacing Fish Meal With Blood Meal On

Chemical Composition Of Supplement For Nile
Tilapia (Oreochromis Niloticus)

J. G. Kirimi, L. M. Musalia & J. M. Munguti

To cite this article: J. G. Kirimi, L. M. Musalia & J. M. Munguti (2016): Effect Of Replacing Fish
Meal With Blood Meal On Chemical Composition Of Supplement For Nile Tilapia (Oreochromis
Niloticus), East African Agricultural and Forestry Journal

To link to this article: http://dx.doi.org/10.1080/00128325.2016.1158898

Published online: 13 Apr 2016.

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 EAST AFRICAN AGRICULTURAL AND FORESTRY JOURNAL, 2016
http://dx.doi.org/10.1080/00128325.2016.1158898

Effect Of Replacing Fish Meal With Blood Meal On Chemical

Composition Of Supplement For Nile Tilapia (Oreochromis
Niloticus)
J. G. Kirimia, L. M. Musaliab and J. M. Mungutic
a
State Department of Fisheries, PO Box 775-60202, Nkubu, Kenya; bDepartment of Animal Sciences, Chuka
University, PO Box 109, Chuka, Kenya; cKenya Marine and Fisheries Research Institute (KMFRI)-Sagana, PO Box
451, Sagana, Kenya
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ABSTRACT KEYWORDS
The objective of the work was to evaluate the effect on the nutrient Amino acids; chemical score;
content of replacing fish meal (FM) with blood meal (BM) in fish essential amino acid index;
supplement. Three isonitrogenous diets (35% crude protein) were nutritive value
formulated using FM as the main source of animal protein (BM0);
50% replacement of FM with blood meal (BM50); or 100%
replacement of FM with BM (BM100). The chemical composition
(ash, crude protein, crude fat and crude fibre) and amino acid
composition were determined. Replacement of FM with BM did
not affect the proximate composition of the diet apart from ash
content which decreased with the level of substitution.
Substituting fish meal with blood meal reduced the levels of
methionine, lysine, isoleucine, leucine, proline, valine and
increased the levels of arginine, phenylanine and alanine in the
diet. Amino acid indices revealed that BM0 had more amino acids
with the highest chemical scores followed by BM50 and BM100. In
all the diets, methionine was the most limiting amino acid. The
essential amino acid index of the diets reduced with the level of
replacement of FM (0.94, 0.88 and 0.77). The study showed
substitution of up to 50% FM with BM gave a useful protein diet
and 100% gave almost a poor protein diet.

Introduction
The nutrient quality of feed ingredients is one of the major prerequisites apart from their
availability for the production of good quality feeds (Sogbesan & Ugwumba, 2008). In view
of the high crude protein level in fish diet, protein is the most significant and expensive
single nutrient in preparation of this diet. Formulating cost effective feeds that meet the
essential amino acid (EAA) requirements of fish and shrimp can be a challenge
(Kaushik & Seiliez, 2010) and depend on relevant data on both EAA requirements of
the fish species and the EAA supplied with the feed. Fish meal has been the main
protein source and the use of alternative feedstuffs requires a thorough understanding
of amino acid requirements and their availability in feedstuffs.

CONTACT Musalia L.M. mugalavai@mail.com

© 2016 KALRO
 2 J. G. KIRIMI ET AL.

Proximate analysis is used in the initial evaluation of feeds and feedstuffs to provide
information on their major nutrient and gross energy contents (Jobling, 2001; Bunda,
et al., 2015). Another evaluation criteria is the protein chemical score (CS) defined as
the lowest ratio of the essential amino acid content in the test protein to the content of
each amino acid in the muscle protein or to the EAA required level when the EAA require-
ment is already established. The assumption of chemical score was that whole egg protein
is of the highest biological value (BV) and therefore the most suitable for growth and that
growth is limited by that essential amino acid in the diet whose ratio to its content in the
whole egg protein is the lowest (Hepher, 1988). Bunda et al. (2015), noted that although
the first limiting amino acid has an important role in determining the relative value of the
dietary protein, it was realized that other essential amino acids may also have some effect
on it and this resulted in the development of the essential amino acid index (EAAI).
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According to Oser (1959), the EAAI is the geometrical mean of the ratio of all EAA in
the evaluated protein relative to their content in a highly nutritive reference protein
such as whole egg. The objective of this study therefore was to investigate the chemical
composition (proximate and essential amino acid composition) of feed ingredients and
the resulting ration when fish meal is replaced with blood meal from local slaughter
houses.

Materials and methods

Study Site
The study was conducted at the National Aquaculture Research, Development & Training
Centre, Sagana, altitude 1230 m above sea level, latitude 0°39´ S and longitude 37°12´ E,
and 90 km north of Nairobi.

Preparation of Diets
The feed ingredients (fish meal, wheat bran, cotton seed cake, soya bean meal and blood
meal) were obtained from the local markets. Bovine blood was collected from the local
Sagana abattoir. Fresh blood drained from freshly slaughtered cattle was collected into a
clean container, transported to Sagana National Aquaculture Centre and boiled immedi-
ately in a cooking container to 100 oC for 45 minutes in order to let the water evaporate
and destroy pathogenic organisms (Khawaja et al., 2007). As the blood boiled, it was con-
tinually stirred until it formed dough. The product was removed from the fire then drained
and crushed manually to increase the drying surface area. The product was spread on a
polythene liner and sun dried for three days to a moisture content of below 15%. The
dried product was milled into a fine powder with a hammer mill.
Three isonitrogenous diets (35% CP) were formulated, in triplicate, using wheat bran,
soybean meal, cotton seedcake and either fish meal as the main source of animal protein
(BM0); 50% replacement of fish meal with blood meal (BM50); and 100% replacement of
fish meal with blood meal (BM100) (Table 1).
The ingredients were ground using a hammer mill to be uniform and mixed thoroughly
by hand in the desired proportion. Water was added to form dough and pelleted using a
pelleter machine to particle size 4.5 mm diameter. The pellets were then dried in the shade.
 EAST AFRICAN AGRICULTURAL AND FORESTRY JOURNAL 3

Table 1. Ingredient composition and calculated chemical composition (%) of fish

supplement containing blood meal as a replacement for fish meal (as fed basis).
BM0† BM50† BM100†
Ingredient
Fish meal 36.6 18.4 0
Blood meal 0 12.5 25.5
Wheat bran 48.4 53.8 59.5
Soybean meal 10 10.3 10
Cotton seed cake 5 5 5
Total 100 100 100
Calculated chemical composition (%)
Crude protein (%) 35 35 35
†BM0, BM50 and BM100 represent replacement of fish meal at 0, 50 and 100%, respectively.

Analysis of Feeds
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The proximate analysis of ingredients and diets were carried out as described by the
AOAC (1995) for crude protein (CP), ether extracts (EE) and ash and crude fibre (CF).
Nitrogen free extracts (NFEs) were estimated by subtracting the total moisture, crude
protein, ether extracts, ash and crude fibre from 100. The method for protein extraction
was adopted from Hamilton et al. (2012) for amino acid analysis using LC-Qtof-MS.
The amino acid score was calculated as:

Chemical score (%) = [essential amino acid of the sample

/essential amino acid of the whole hen egg] × 100

While essential the EAAI was calculated as:

EAAI = n/{(aa1 /AA1 )(aa2 /AA2 ).(aan /AAn )}

Where EAAI is the nth root of the essential amino acids in the test diet (aa) to the content
of each of those amino acids in the reference tissue (AA) and n is the total number of
amino acids evaluated (Tidwell et al., 1993). Measurements were done in triplicate. The
data were subjected to one-way analysis of variance (ANOVA) using Statistical Package
for Social Sciences version 17.0 (SPSS Statistics) and where there were differences,
mean separation was done by least significant difference (LSD).

Table 2. Proximate composition of feed ingredients (%) used to formulate fish supplements
containing blood meal as a replacement for fish meal.
Fish meal Blood meal Soybean meal Cotton seed cake Wheat bran
Proximate composition (%)
Dry matter 90.85c±0.24 89.47e±0.06 92.31a±0.07 92.06ab±0.17 89.55de±0.07
Crude protein 64.20b±0.27 80.41a±0.13 11.47e±0.04 27.07c±0.40 14.49d±0.02
Ether extracts 5.07c±2.14 0.62e±0.21 5.85bc±0.44 8.52ab±0.40 2.13abcde±0.33
Ash 16.13a±0.19 4.52b±0.31 3.07c±0.23 3.92bcd±0.89 5.78be±0.73
Crude fibre 0.52e±0.12 1.34de±0.28 9.75c±0.87 23.43a±0.77 12.58b±0.85
Nitrogen free extracts 4.94d±1.63 2.62de±0.28 62.17a±0.47 29.14c±0.66 54.6b±1.14
Values are expressed as mean ± SE.
a,b,c,d,e
Values in the same row having different superscript letters are significantly different (P <0.05).
 4 J. G. KIRIMI ET AL.

Results
Chemical Composition of Diets
The proximate composition of the feed ingredients is shown in (Table 2). Blood meal
recorded the highest crude protein content of 80.41% with soy bean meal recording the
lowest crude protein content of 11.47%. Cotton seedcake had the highest crude fibre
content (23.43%) with fish meal recording the lowest crude fibre content (0.52%). The
ash content was high in fish meal (16.13%) compared with soybean meal (3.07%).
Blood meal had a low residual oil compared to the cotton seedcake (0.62% vs 8.5%).
The proximate composition of the formulated diets is shown in Table 3. Replacement of
FM by BM increased (P<0.05) the crude protein values. Ether extracts were the same for
BM50 and BM100 with a slight increase in BM0 (3.4%). Total replacement of fish meal
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had the lowest ash content with BM0 recording almost twice that of BM100. However,
BM0 recorded the lowest crude fibre content (9.3%) with BM50 and BM100 recording
almost the same figure.

Amino Acid Composition

Blood meal had the highest amount of isoleucine (118.07 mg/g protein) although phenyl-
alanine combined with tyrosine had 137.01 mg/g protein (Table 4). Alanine was the lowest
(13.97) followed by arginine and leucine. The chemical score was highest for phenyl-
alanine and tyrosine (147.32%), methionine (71.91%) and lysine (64.06%).
Methionine, leucine, isoleucine, valine and proline contents of BM100 were lowest and
highest in BM0. Arginine was high in BM100 with BM50 and BM0 being almost the same.
BM50 recorded the highest content of tyrosine and lysine compared to the other diets
(Table 5).
Data on chemical scores and EAAI (Table 6) shows that isoleucine had the highest per-
centage chemical score in the three supplements, i.e. 178.20, 142.31 and 136.81 for BM0,
BM50 and BM100, respectively. However, the chemical score percentage for methionine
was lowest in BM100 with BM0 recording the highest score. The chemical score for
lysine decreased with the level of substitution of FM with BM (85.41, 68 and 64.83).
BM0 had the highest chemical scores for methionine, leucine, isoleucine and valine
while BM100 recorded the lowest chemicals scores for methionine, valine, isoleucine,
leucine and phenylalanine+ tyrosine. The EAAI was highest in BM0 (0.94) followed by
BM50 (0.88) and BM100 (0.77)

Table 3. Proximate composition of fish supplements containing blood meal as a

replacement for fish meal.
BM0† BM50† BM100†
Dry matter 90.59c±0.10 91.15b±0.13 91.37ab±0.08
Crude protein 32.08c±0.15 33.88a±0.39 33.69ab±0.03
Ether extracts 3.41a±0.35 3.00a±0.28 3.00a±0.40
a b
Ash 9.45 ±0.10 7.13 ±0.07 5.1c±0.58
Crude fibre 9.3c±0.23 11.37a±0.20 11.28ab±0.36
Nitrogen free extracts 36.36b±0.25 35.78b±0.49 38.3a±0.66
Values are expressed as mean ± SE.
a,b,c
Values in the same row having different superscript letters are significantly different (P <0.05).
†BM0, BM50 and BM100 represent replacement of fish meal at 0, 50 and 100%, respectively.
 EAST AFRICAN AGRICULTURAL AND FORESTRY JOURNAL 5

Table 4. Amino acid composition (mg/g protein) and chemical score of blood meal.
Amino acid Composition Chemical score
Methionine 40.99 71.91
Lysine 44.84 64.06
Leucine 30.03 34.92
Isoleucine 118.07 218.65
Valine 61.43 93.08
Phenylalanine+tyrosine 137.01 147.32
Alanine† 13.97
Arginine 23.45
Proline† 39.01
†Non-essential amino acids.

Discussion
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Proximate Composition
The proximate composition of the test ingredients, i.e. fish meal, cotton seed cake, wheat
bran and blood meal used in this study was within the range of values reported by other
authors (Drew et al., 2007; Um-E-Kalsoom et al., 2009; Al Mahmud et al., 2012). The
crude protein (11%) was markedly low in soybean meal. Lovell (1988) noted that the nutri-
ent composition of feedstuffs depends on the origin, state and processing methods used.
However, according to the National Research Council (1993) solvent extraction of the oil
results in soybean meal (SBM) containing 44% crude protein if the soybean hulls are
included or 48% crude protein without the hulls. The low CP content was due to adultera-
tion of the soybean meal by marketing agents using cheap low quality ingredients like
sawdust. This was reflected in the high CF content of SBM (9.75%) which was more
than double that recorded by Agbo (2008) and Noreen and Salim (2008), i.e. 3.82% and
1.09%, respectively. It is important to note that although the crude protein level of
soybean was below the expected level, this had the same effect across the three diets for-
mulated because inclusion levels of soybean meal were the same at 10%. This reveals that
farmers can purchase adulterated ingredients leading to the formulation of substandard
feeds which in turn is reflected by the poor performance of the animal.
Cottonseed cake recorded a higher crude fibre of all the ingredients (Table 2) which is
considered a limiting factor in its use as feed. Nagalakshmi et al. (2007) observed that a

Table 5. Amino acid composition (mg/g protein) of fish supplements containing blood meal
as a replacement for fish meal.
Amino acid BM0† BM50† BM100†
Methionine 38.31a±0.08 35.11a±2.39 30.09a±3.00
Lysine 45.38a±14.35 59.79a±3.01 47.60a±9.50
Leucine 75.62a±1.40 61.43b±2.24 47.18c±1.35
Isoleucine 96.23a±0.20 76.85a±10.43 73.88a±11.07
Alanine 33.54a±4.82 30.89a±7.44 46.62a±8.99
Valine 62.06a±2.54 50.44b±0.52 46.34c±0.78b
Arginine 20.87a±0.85 20.03a±0.88 24.05a±1.58
a b
Proline 57.09 ±2.24 47.15 ±0.51 45.19bc±2.12
Phenylalanine 71.36a±7.42 70.48a±1.00 74.04a±3.30
Tyrosine 27.31a±3.73 34.09a±1.76 27.93a±1.69
Values are expressed as mean ± SE.
a,b,c
Values in the same row having different superscript letters are significantly different (P <0.05).
†BM0, BM50 and BM100 represent replacement of fish meal at 0, 50 and 100%, respectively.
 6 J. G. KIRIMI ET AL.

Table 6. Chemical scores (%) and essential amino acid index (EAAI) of fish supplements
containing blood meal as a replacement for fish meal.
BM0† BM50† BM100†
Amino acid Chemical score
Methionine 67.21a±0.14 61.59a±4.19 52.79a±4.64
Lysine 64.83a±20.50 85.41a±4.30 68.01a±13.57
Leucine 87.93a±1.60 71.43b±2.62 54.86c±1.56
Isoleucine 178.20a±0.37 142.31a±19.31 136.81a±20.49
Valine 94.03a±3.84 76.42b±0.79 70.21bc±1.17
Phenylalanine + tyrosine 106.08a±11.99 112.44a±2.97 109.61a±1.73
Essential amino acid index 0.94a±0.60 0.88a±0.35 0.77a±0.20
Values are expressed as mean ± SE.
a,b,c
Values in the same row having different superscript letters are significantly different (P <0.05).
†BM0, BM50 and BM100 represent replacement of fish meal at 0, 50 and 100%, respectively.
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high level of crude fibre in cotton seed cake is inversely proportional to the concentration
of protein and further revealed that the crude protein of undecorticated cotton seed meal
ranged from 22.2 to 30.31%.
The crude protein content of fish meal (64.20%) was below that obtained by Otubusin
(2009) who recorded 70% CP. This figure together with the ash content (16.3%) were
within the normal range which according to Drew et al. (2007), may vary from 50 to
70% and 10 to 21%, respectively, depending on fish species, the source and processing
method. The crude protein of wheat bran was the same as that obtained by
Um-E-Kalsoom (2009) and Al Mahmud et al. (2012) but ash and ether extracts were rela-
tively low. The crude protein of blood meal (80%) was close to the results by Drew et al.
(2007) while the ash content was double (4.5%). However, Otubusin et al. (2009) recorded
similar results for crude protein but ash content was zero.
The proximate analysis of the three supplements (Table 3) shows that the crude protein
had slight variation from the formulated diet on as fed bases (Table 1). This was due to
fluctuation in the crude protein content of the ingredients (Table 2), in particular
soybean meal and cottonseed cake. Substitution of FM with BM gave a higher crude
fibre content of 11.37% for diet BM50 and 11.28% for BM100. This was due to increased
amounts of wheat bran incorporated in the diet to adjust for the CP content in the formu-
lation of the diets (Table 1). A high fibre content reduces the total dry matter and nutrient
digestibility of the diet, resulting in poor performance (De Silva & Anderson, 1995). In
addition it adds to the faecal waste which affects the water quality and hence fish perform-
ance (Lovell, 1998). According to De Silva and Anderson (1995), crude fibre was within
the normal range of 8–12% for diets of fish.
A high ash content of BM0 (9.45%), which was almost double when 100% of FM was sub-
stituted with BM, was due to the high ash content in fish meal (Table 2). The FM is usually
dried on sandy ground along the lake, which contributes to the high ash content. There was
not much variation in the lipid content (3–3.4%) with the substitution of FM. However, this
was below the recommended levels of 5–12% for tilapia (Suresh, 2003).

Amino Acid Composition

Low methionine content on substitution of FM was due to low methionine in blood meal
compared to fish meal. Similarly, substituting fish meal wholly with blood meal, as in the
 EAST AFRICAN AGRICULTURAL AND FORESTRY JOURNAL 7

case of BM100, reduced the level of some amino acids in the supplement, i.e. methionine,
lysine, isoleucine, leucine, proline, valine and increased arginine, phenylanine and alanine
in the supplements. Thus, BM0 displayed a better amino acid profile with higher levels of
methionine, isoleucine, leucine, lysine, valine and proline.
It is worth noting that cystine was not detected in the supplements (Table 5) and
according to the National Research Council (1993) there exists a relationship among
amino acids such that cystine can be formed metabolically from dietary methionine at
a rate sufficient to meet the requirements of fish but the reverse sequence of reactions
does not occur. Methionine can thus meet the total sulphur amino acid requirement of
fish, although some of this requirement may be met by cystine (National Research
Council, 1993). Based on this, the cystine requirement for fish can be met by the methion-
ine content in the diet. In addition, phenylalanine and tyrosine were present in the three
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supplements but a similar relationship exists between aromatic amino acids (phenyl-
alanine and tyrosine). Fish readily convert phenylalanine to tyrosine so that phenylalanine
alone can meet the requirements for aromatic amino acids but the presence of tyrosine in
the diet reduces some of the requirement for phenylalanine (National Research Council,
1993).
Considering the amino acid profile in the supplements against the recommended levels
for Nile tilapia (National Research Council, 1993), the amino acid profiles were low which
can lead to poor utilization of the dietary protein and consequently reduce growth and
decrease feed efficiency (Halver & Hardy, 2002). Dietary amino acid utilization requires
that all amino acids are simultaneously present in adequate concentrations at sites of
protein synthesis. Hence, deficiency of an essential amino acid limits protein synthesis
to the level of that particular essential amino acid, the remainder being catabolized
(Sveier et al., 2001). For most essential amino acids, deficiency translates to a reduction
in weight gain. In some species of fish, a deficiency of methionine or tryptophan leads
to pathologies, because these amino acids are not only incorporated into proteins but
also used for the synthesis of other compounds (Lovell, 1998). For example, cataracts
occur in salmonids and rainbow trout as a consequence of methionine (sulphur amino
acids) and tryptophan deficiency, respectively, in their diets (Lovell, 1998). The amino
acids balance in the supplements for the present study were not met because they were
not formulated based on an ideal protein concept and according to Yamamoto et al.
(2004), the diets could depress feed intake and growth of fish.

Chemical Score and Essential Amino Acid Index

In the present study (Table 4), BM as an ingredient displayed a high chemical score for
phenylalanine + tyrosine and isoleucine. The most limiting amino acid in BM100 and
BM50 was methionine. This could be attributed to the use of blood meal. In the
present study, high chemical scores were associated with low substitution of fish meal,
which is better balanced in amino acids than blood meal.
Although the chemical score is important in determining the relative value of dietary
protein, other essential amino acids could also have an effect on the nutritive value of
the dietary protein, as reflected in the EAAI (Table 6). The chemical score is based on
the assumption that whole egg protein is of the highest biological value and thus the
most suitable for growth, which could be limited by the EAA in the diet whose ratio to
 8 J. G. KIRIMI ET AL.

its content in the whole egg protein is the lowest (Hepher, 1988). The EAAI of the three
supplements, BM0, BM50 and BM100 was estimated to be 0.94, 0.88 and 0.77, respect-
ively. Good quality protein sources have an EAAI greater than or equal to 0.90, useful
protein sources have a value of 0.80 whereas sources with values below 0.70 are considered
to be inadequate (Oser, 1959; Penaflorida, 1989). Thus, BM0 in the present study could be
considered to be a good quality protein supplement, BM50 a useful protein supplement
and BM100 was closer to a poor protein source.

Conclusion
The proximate contents of the three supplements were within the required level for
growing fish. However, considering the amino acid composition, substitution of up to
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50% FM with BM gave a useful protein diet and 100% gave almost a poor protein diet.
Based on this, it can be concluded that blood meal can partially replace 50% fish meal
in the diets of Oreochromis niloticus. Further studies on the use of blood meal as a replace-
ment for fish meal on the nutritive value of diets for Oreochromis niloticus is
recommended.

Funding
The authors thank Kenya Marine and Fisheries Research Institute (KMFRI) for funding the analysis
of this work. Special thanks goes to International Center for Insect Physiology and Ecology (ICIPE)
and Department of Animal Sciences, Egerton University for providing their facilities for proximate
and amino acid analysis.

ORCID
L. M. Musalia http://orcid.org/0000-0003-3709-1754

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