Download Effect of Natural Fermentation on the Vitamins, Amino Acids and

AMERICAN JOURNAL OF FOOD AND NUTRITION
Print: ISSN 2157-0167, Online: ISSN 2157-1317, doi:10.5251/ajfn.2016.6.3.91.100
© 2016, ScienceHuβ, http://www.scihub.org/AJF
Effect of Natural Fermentation on the Vitamins, Amino Acids and Protein
Quality Indices of Sorghum-Based Complementary Foods
Sengev, I. A*., Ariahu, C. C. and Gernah, D. I.
Department of Food Science and Technology, University of Agriculture, P.M.B. 2373,
Makurdi, Benue State, Nigeria
*
Corresponding author: Sengev, I. A.
Department of Food Science and Technology, University of Agriculture, P.M.B. 2373,
Makurdi, Benue State, Nigeria, E–mail: [email protected] Tel: +2348035020615
ABSTRACT
This study investigated the effect of natural fermentation on the vitamins, amino acid and the
quality of protein of sorghum-based complementary foods. Samples were formulated based on
16% protein to satisfy the nutrient requirement of preschool children to obtain non-fermented
sorghum/mango mesocarp/crayfish (NFSMF), non-fermented sorghum/fluted pumpkin leaf/fish
(NFSPF), fermented sorghum/ mango mesocarp/crayfish (FSMF) and fermented sorghum/ fluted
pumpkin leaf /crayfish (FSPF). The vitamins, amino acids and growth study parameters were
examined using standard procedures. Vitamin A (193.50 - 322.10μ/100g), B12 (0.48 –
0.54mg/100g) and D (0.42 – 0.93mg/100g) increased significantly (p< 0.05) with fermentation,
both for samples containing mango mesocarp and fluted pumpkin powders while retinol
equivalent decreased with fermentation. All the samples met the essential amino acids
requirement as recommended by WHO/FAO/UNU pattern 2007. All the essential amino acids
increased significantly with fermentation. NFSMF and NFSPF exhibited higher growth response
when fed to rats compared to FSMF and FSPF. Sample NFSMF gave better protein efficiency
ratio (PER) (2.48) and net protein ratio (NPR) (4.12) followed by NFSPF, FSMF and FSPF.
Among all the samples, NFSMF gave the best PER result.
Key words: Sorghum, Vitamins, Crayfish, Mango, amino acids
INTRODUCTION
According to the recent FAO database, the total
number of undernourished people in the world,
almost 794.6 million, is still unacceptably high, and
eradication of hunger remains a major global
challenge (FAO/IFAD/WFP, 2015). Hence, the
search for alternative food ingredients remains of
utmost importance.
In Nigeria, traditional complementary foods are
usually produced from staple cereals and legumes
prepared either individually or as composite gruels
(Walker, 1990). Cereal grains are considered to be
one of the most important sources of dietary proteins,
carbohydrates, vitamins, minerals and fibre for
people in developing countries. Complementary
feeding period is the time when malnutrition starts in
many infants, contributing significantly to the high
prevalence of malnutrition in children less than 5
years of age worldwide (Daelmans and Saadeh,
2003). The introduction of complementary food in
Nigeria usually starts between age 4 and 6 years;
and usually involves the use of a semi-liquid porridge
prepared locally by the mother from staple cereals or
tubers (Bentley et al., 1991), (Nout, 1993), (Guptill et
al., 1993). Combination of common cereals, which
are deficient in lysine but have sufficient amount of
sulphur-containing amino acids, with inexpensive
animal protein sources like crayfish that are rich in
lysine can be used to improve the nutritive value of a
food product.
Sorghum variety (Sorghum bicolor [L.] Moench) is
one of the most important staple foods in low-income
and high-income countries ( Shehab et al., 2010).
The nutrient composition of sorghum indicates that it
is a good source of energy, proteins, carbohydrates,
vitamins and minerals including the trace elements,
particularly iron and zinc, except calcium. Sorghum
grain contains minerals such as phosphorus,
potassium and magnesium in varying quantities
(Dicko et al., 2006).
Wet processing including
soaking, germination and fermentation leads to a
reduction in phytic acid and increases of the minerals
solubility in foods and could thus improve
bioavailability of minerals in cereals and legumes
(Afify et al., 2011a). The low digestibility is due to
protein-protein, protein-carbohydrate, protein (poly)
91
Amer. J. Food & Nutr., 2016, 6(3): 91-100
o
phenol and carbohydrate- (poly) phenol interactions
(Knudsen et al., 1988). The choice of crayfish is
based on the fact that it has high protein in range of
60 to 70% (Obizoba, 1985) and its inclusion in
sorghum-based foods would improve on the protein
quality. Mango pulp is reported to be rich in
micronutrients (Belly, 2006) and fluted pumpkin was
reported to be rich in amino acids and has haematinic
properties (Fasuyi, 2006), (Dina et al., 2006).
FAO/WHO/UNICEF (1971) emphasised the use of
local foods formulated at home and guided by the
following principles: (i) high nutritional value to
supplement breastfeeding, (ii) acceptability, (iii) low
price, and (iv) use of local food materials (Dewey and
Brown, 2003), (Pelto et al., 2003). Therefore, the aim
of this study was to develop and evaluate the
nutritional properties of the complementary foods
using sorghum, crayfish, mango mesocarp and fluted
pumpkin powders.
MATERIALS AND METHODS
dried in hot air oven at 70 C for 12h and milled into
fine flour of 0.5mm instead of 0.212mm.
Preparation of Mango Mesocarp Powder: The
method of Sengev et al. (2010) was adopted with
modification. Five kilograms of partially ripe mango
fruits, chul kpev (a local variety) (pH = 3.8, Brix = 7.0,
Refractive Index = 1.34) were sorted, washed, peeled
and the mesocarp was manually sliced to an average
thickness of 2.5mm. The slices were spread on a tray
covered with aluminum foil and oven-dried at 70 
o
1 C for 24h to a moisture content of about 10%. The
slices were milled using a disc attrition mill (Model:
AII Asiko, Nigeria) and sieved through a 0.5mm sieve
to obtain mango mesocarp flour (MMF).
Preparation of Crayfish Powder: The method of
Onuorah and Akinjede (2004) was adopted with
modification. About 1kg of crayfish was washed to
remove extraneous materials, sundried for 12h at
relative humidity of 65%. The sample was milled
using hammer mill (Model: Brook Crompton Series
2000, England) and sieved through 0.5mm laboratory
test sieve instead of 0.6mm to obtain crayfish flour.
Sample Procurement: About 10kg of red sorghum
grains [Sorghum bicolor, (L) Moench] and 5kg of
semi ripe mango fruits (a local variety) (Mangifera
indica) popularly known as Wua nyian and Chul kpev
in Tiv respectively, 1kg of crayfish (Procambarus
clarkii) and fluted pumpkin leaves (Telferia
accidentalis) each were sourced from a local market
in Makurdi, Benue State. Vitamin pre-mix (Vitalyte)
was also sourced from a pharmacy store in Makurdi.
These materials were transported to the Department
of Food Science and Technology, University of
Agriculture, Makurdi for processing prior to product
formulation and subsequent analysis. Male wistar
strain weanling albino rats (4 weeks old) were
sourced from National Institute of Trypanasomiasis
Research, Vom, Plateau State.
Preparation of Sorghum Flour: About 10kg of
sorghum was dehulled using rice huller (Model:
Navin, Madras) and washed with tap water and sun
dried for 12h at average relative humidity of 65%.
The dehulled grains were milled using attrition mill
(Model: Asiko AII). The flour was sieved using a
laboratory test sieve of 0.5mm aperture.
Solid State Fermented of Sorghum Flour:
Fermentation of sorghum flour was carried out using
the method of Sengev et al. (2010) with modification.
The milled sorghum flour was divided into two equal
parts. One part was mixed with tap water in the ratio
o
of 2:1w/v and allowed to ferment for 48h at 30  1 C,
relative humidity of 65% in none air-tight covered
plastic tray. At the end of the fermentation, a pH of
3.80 was recorded. The fermented sorghum was
Preparation of Fluted Pumpkin Leaf Powder: The
flute pumpkin leaves were washed with tap water,
steam blanched for 3 sec. and dried under the shade
to constant weight. The dried leaves were milled
using attrition mill (Model: AII Asiko, Nigeria) and
screen through a 0.5mm sieve to obtain fluted
pumpkin leaf flour.
Blend formulation: The blends were formulated
based on protein and mass balance equations.
Sorghum, crayfish, mango mesocarp and fluted
pumpkin leaf powders were blended based on 16%
protein level as shown in Table 1.
Feeding trial Studies: Feeding trial studies were
conducted using 21 weanling male albino rats (Wistar
strain), that were clinically healthy (average weight of
90.10g), using a modification of the method
described by Rasaco (2002). A complete randomized
design (CRD) was used in which the rats were
initially weighed to the nearest 0.1g at the start of
each feeding trial and allocated into the seven cages
based on weight equivalent with 3 rats per cage. The
cages were placed on ceramic tiles to permit
collection of faeces. The six diets formulated
according to Pellet and Young (1980) along with the
basal diet (Table 2) were evaluated. The diets were
administered to the rats based on their body weight
(ie 10g food per 100g body weight) and water adlibitum for 28 days. The total food intake of the rats
was determined by recording the food left after daily
92
Amer. J. Food & Nutr., 2016, 6(3): 91-100
intake. Daily weight gain was obtained by weighing
all the rats individually. Protein consumption was
calculated from the food intake. All faeces collected
in the last 10 days of feeding were pooled together,
dried, weighed and milled into fine powder for
analysis.
amino acid values reported were the averages of two
determinations. Norleucine was the internal standard.
Statistical Analysis: The data generated were
statistically analysed using analysis of variance
(ANOVA). Means were tested for significant
difference using Duncan’s Multiple Range Test
(DMRT) (Wahua, 1999). Significance level was
accepted at p < 0.05.
RESULTS
Vitamin Content and Retinol Equivalent of
Sorghum-Based Complementary Foods: The
results of vitamin concentration and retinol equivalent
of sorghum-based complementary foods are
presented in Table 3. The results revealed that
significant difference (p<0.05) existed between the
samples. Retinol equivalent ranged from 120.70 –
202.60 RE, Vitamin A ranged from 193.50 – 322.10
μg/100g, Vitamin C ranged from 11.30 – 12.96
mg/100g, Vitamin B1 ranged from 0.87 – 0.94
mg/100g, Vitamin B6 ranged from 0.43 – 0.55
mg/100g, Vitamin B9 ranged from 0.13 –
0.14mg/100g, Vitamin B12 ranged from 0.48 – 0.53
mg/100g and Vitamin D ranged from 0.42 – 0.93
mg/100g for NFSMF, NFSPF, FSMF and FSPF.
Amino Acid Composition of Sorghum–Based
Complementary Foods: The essential and non
essential amino acid compositions of sorghum
crayfish based complementary foods are presented
in Table 4 and 5 respectively. The essential amino
acids were compared with 2007 WHO/FAO/UNU
report. For the essential amino acids, lysine ranged
from 49.40 to 58.80mg/g protein, threonine ranged
from 36.20 to 44.61mg/g protein, valine ranged from
35.15 to 44.20mg/g protein, methionine values
ranged from 19.45 to 24.79mg/g protein. Others
include isoleucine which ranged from 27.45 to
35.05mg/g protein, leucine ranged from 69.03 to
78.40mg/g protein and phenylalanine ranged from
39.55 to 45.89mg/g protein. Tryptophan ranged from
1.62 to 2.06mg/g protein. The essential amino acids
met the WHO/FAO/UNU 2007 pattern.
Protein Quality Evaluation: Net Feed intake and
body weight achieve were employed to determine
growth study parameters such as protein efficiency
ratio (PER), relative protein efficiency ratio (R-PER),
net protein ratio (NPR) and relative net protein ratio
(R-NPR). Others include feed conversion factors
such as feed conversion ratio (FCR) and feed
conversion efficiency (FCE) following the procedure
described by Shiriki et al. (2015).
Analyses
Vitamin Analysis: Vitamin A, B1, B6, B9, B12 and D
were evaluated using HPLC (Model: BLC-10/11,
BUCK Scientific, USA) techniques as described by
AOAC (2005). For each sample, 3.0g was mixed
with 5mL of n-hexane and 20mL of HPLC grade
water. The mixture was homogenized at 12000 rpm
and centrifuged (3500 x g) for 30 min. followed by
sequential filtration through whatman No 1 filter paper
and 0.45μm membrane. Then 15μL of the
supernatant was injected into the HPLC equipped
with a UV detector set at 254nm. The peaks of the
vitamins in the samples were calculated in relation to
the peaks of standard vitamins.
Determination of Vitamin C Using Direct Tittration
Method: Vitamin C content was determined
according to Suntornsuk et al. (2002) by direct
titration with iodine solution (Potassium iodide +
Iodine). Ten gram (10g) of the flour was mixed with
100mL of distilled water in 250mL beaker and
allowed to stand for 30min. The extract (40mL) was
transferred into a 250mL volumetric flask and 2 drops
of 0.5% starch indicator solution was added. The
mixture was titrated with 0.0057M solution of iodine
previously standardized with ascorbic acid solution.
Vita min C (mg / 100 g ) 
Vita min C concentration 100
x
Volume of extract
1
For the non-essential amino acids, histidine ranged
from 18.00 to 19.56 mg/g protein, arginine ranged
from 60.80 to 75.36 mg/g protein, aspartic acid
ranged from 77.47 to 88.45 mg/g protein, serine
ranged from 30.06 to 38.92 mg/g protein, glutamic
acid ranged from 84.97 to 97.86 mg/g protein, proline
ranged from 24.92 to 34.62 mg/g protein. Others
include glycine which ranged from 38.83 to 48.96
mg/g protein, alanine ranged from 29.87 to 34.77
mg/g protein, cystine ranged from 11.95 to 15.77
mg/g protein and tyrosine ranged from 22.21 to 29.79
mg/g protein.
Amino Acid Analysis : Amino acid analysis was by
ion exchange chromatography (IEC) [FAO/WHO,
1991] using the Technicon Sequential Multisample
(TSM) Amino Acid Analyser (Technicon Instruments
Corporation, New York). The period of analysis was
76 min for each sample. The gas flow rate was 0.50
mL/min at 60°C with reproducibility consistent within
±3%. The net height of each peak produced by the
chart recorder of the TSM (each representing an
amino acid) was measured and calculated. The
93
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Table 1: Formulation of Blends
Blend
Ingredient mix (g/100g)
Sorghum
Mango Mesocarp Powder
Fluted Pumpkin Leaf Powder
Crayfish Powder
NFSMF
91.06
0.17
-
8.77
NFSPF
91.04
-
0.19
8.77
FSMF
91.06
0.17
-
8.77
FSPF
91.04
-
0.19
8.77
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF =
Fermented Sorghum + Mango Mesocarp + Crayfish, FSPF =Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish.
Table 2: Formulation of Experimental Diets
Ingredient
Formulated Diet
NFSMF
NFSPF
FSMF
FSPF
WSF
Basal Diet
Milk
Blend
57.78
56.06
58.52
57.28
Corn Starch
22.22
23.94
21.48
22.72
80.00
43.32
Milk Powder
36.68
Sorghum Flour
80.00
Vegetable Oil
10
10
10
10
10
10
10
Rice Husk
5
5
5
5
5
5
5
Salt
4
4
4
4
4
4
4
Vitalyte
1
1
1
1
1
1
1
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF =
Fermented Sorghum + Mango Mesocarp + Crayfish, FSPF =Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, WSF = Whole Sorghum Flour
Table 3: Effect of Natural Fermentation on the Vitamin Content of Sorghum-Based Complementary Foods
Sample
*Retinol
Vitamin A
Vitamin C
Vitamin B1
Vitamin B6
Vitamin B9
Vitamin B12
Vitamin D
Equivalent
(μg/100g)
(mg/100g)
(mg/100g)
(mg/100g)
(mg/100g)
(mg/100g)
(mg/100g)
a
c
ab
b
d
b
c
c
NFSMF
202.60±0.39
224.80±0.27
12.46±0.01
0.87±0.03
0.43±0.00
0.13±0.02
0.51±0.01
0.61±0.01
b
a
a
b
c
b
b
b
FSMF
169.60±0.27
322.10±0.21
12.96±0.20
0.88±0.00
0.48±0.00
0.13±0.00
0.53±0.00
0.77±0.00
c
d
b
a
a
a
d
d
NFSPF
124.30±0.37
193.50±0.03
11.30±0.41
0.94±0.00
0.55±0.00
0.14±0.04
0.48±0.01
0.42±0.00
d
b
ab
b
b
a
a
a
FSPF
120.70±0.32
306.60±0.46
11.70±0.82
0.89±0.00
0.53±0.00
0.14±0.01
0.54±0.00
0.93±0.00
-3
RDA
400
400
25
0.6
0.6
0.2
1.2x10
0.02
LSD0.05
0.94
0.61
1.30
0.03
0.01
0.01
0.01
0.01
Values are mean ± Standard deviation of three determinations. Means in the same column not followed by the same superscript are significantly different (p
< 0.05)
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF =
Fermented Sorghum + Mango Mesocarp + Crayfish, FSPF = Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, *Conversion factor based on
6μg/100g β-Carotene: 1 Retinol Equivalent, RDA = Recommended Daily Dietary Allowance from IOM (2005)
94
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Growth Response of Rat Fed with SorghumBased Complementary Food: The results of growth
response of rats fed with sorghum and crayfishbased complementary foods for 28 days are shown in
Figure 1. In the rat bioassay, all rats survived to the
end of the observation and gained positive body
weight except for zero protein group. The test diets
fed to the animals include NFSMF, NFSPF, FSMF
and FSPF. Others include WSF as control, basal diet
as non-protein diet and milk as the standard or
reference diet. The increase in body weight was
reported as 52.82g (40.56%) for NFSMF, 44.20g
(35.16%) for NFSPF, 35.57g (28.03%) for FSMF and
14.35 (14.66%) for FSPF. Others include 5.95g
(6.22%) for WSF 53.50g (39.41%) for milk while the
basal diet recorded -35.03 (40.45%) reduction in
body weight during the 28-day feeding trial.
blends could be ascribed to the contributions from
mango mesocarp and fluted pumpkin leaf. Thiamin is
responsible for the metabolism of carbohydrates and
branched-chain amino acids. The results of this study
did not show any evidence of increase in thiamin
during fermentation as reported by Kazanas and
Field (1981) probably due to short fermentation time.
Dicko et al. (2006) again reported 0.35mg/100g
thiamin for sorghum grain lower than values reported
in this study. The vitamin B6 (Pyridoxine) in the
blends was lower but compared favourably with the
report of Duodu et al. (2003) and Dicko et al. (2006)
who reported 0.50mg/100g pyridoxine. Vitamin B6 is
responsible the metabolism of amino acids and
glycogen in the body. The low values could be
attributed to the dehulling which removed the pericap,
aleurone layer and the germ leading to reduction
vitamin B6 concentration. The
vitamin B6
concentration in the samples fell slightly below the
recommended dietary allowance of 0.6mg/day as
reported by IOM (2005). The vitamin B9 (Folic acid)
did not show significant difference between the
fermented and non-fermented samples. However, the
blends provided at least 65% of the RDA (IOM,
2005). Folic acid is important for many metabolic
reactions, in the biosynthesis of DNA and RNA and
the inter-conversions of amino acids. Also, folate
possesses antioxidant competence that protects the
genome by preventing free radical attack (LeBlanc et
al., 2011).
Protein Quality Evaluation of Sorghum-CrayfishBased Complementary Foods: The growth study
parameters of rats fed for 28 days with sorghumcrayfish-based complementary foods and the
reference diet (Standard) are presented in Table 5.
The food conversion ratio (FCR) ranged from 3.80 to
30.15, food efficiency conversion (FCE) ranged from
3.33 to 26.34%. Similarly, the protein efficiency ratio
(PER) for the test diets ranged from 0.47 to 2.68, the
relative protein efficiency ratio (R-PER) ranged from
0.44 to 2.50, net protein ratio (NPR) ranged from 2.74
to 4.44 while relative net protein utilization (R-NPR)
ranged from 2.48 to 4.02.
The vitamin B12 concentration of the blends increased
significantly (p<0.05) with fermentation. The increase
in vitamin B12 could be attributed to activities of
microorganisms during solid state fermentation of
sorghum flour. Murdock and Fields (1984) reported
that the levels of vitamin B12 and folic acid were
increased by lactic acid fermentation of maize.
Mustafa et al. (2013) also reported the production of
vitamin B12 by Klebsiella pneumonia during solid state
fermentation of agricultural waste. The abundance of
vitamin B12 implies that prospective consumers would
not suffer from pernicious anaemia. Vitamin B12 is
required for the metabolism of fatty acids, amino
acids, nucleic acids and carbohydrates (QuesadaChanto et al., 1994).
DISCUSSION
Vitamin Content and Retinol Equivalent of
Sorghum-Based Complementary Foods: The
retinol (RE) content of the blends decreased
significantly (p<0.05) with fermentation while the
vitamin A (retinol) content increased. It is postulated
that the observed increase in vitamin A may be
ascribed to its synthesis by microorganism during
fermentation. The summation of retinol activity (REA)
i.e RE+vitamin A met the RDA recommended by IOM
(2005) for all the samples except for NFSPF which
delivered 317.80 μg/100g (79.45%) REA. The
occurrence of vitamin A in the blend may be ascribed
to the inclusion of crayfish in the diets. Hamdi (2011)
reported 810μg/100g vitamin A in Procambarus
clarkii . The inclusion of fluted pumpkin and mango
mesocarp
powders
increased
vitamin
C
concentration in the blends. The vitamin C
concentration in the blends was higher than the value
reported for plain sorghum. Dicko et al. (2006)
reported <0.001mg/100g vitamin C for plain sorghum.
The higher values of vitamin B1 (thiamin) in the
Fermentation significantly (p<0.05) increased the
concentration of vitamin D. The high amount of
vitamin D in the blends would meet the high demand
of the vitamin for skeletal growth in infant.
95
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Fig. 1: Effect of natural fermentation on the growth response of rats fed with sorghum-based complementary foods
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF =
Fermented Sorghum + Mango Mesocarp + Crayfish, FSPF =Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, WSF = Whole Sorghum Flour
96
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Table 4: Essential Amino Acid Composition of Sorghum--Based Complementary Foods
Essential Amino Acid WHO/FAO/UNU
Sample
LSD0.05
(mg/g protein)
Pattern* 2007
NFSMF
FSMF
NFSPF
FSPF
Lysine
39
49.40±1.13b
57.10±1.27a
52.10±1.13b
58.80±0.42a
2.90
Threonine
20
36.20±1.27c
39.60±0.99b
41.45±0.92b
44.61±0.68a
2.74
Valine
33
35.15±1.20c
41.00±0.28b
40.25±0.92b
44.20±1.56a
3.04
Methionine
19
19.45±0.21d
21.46±0.08c
22.81±0.12b
24.79±0.59a
0.89
Isoleucine
25
27.45±0.21d
31.05±0.35b
29.08±0.75c
35.05±0.21a
1.22
Leucine
49
71.17±0.25b
72.46±0.08b
69.03±0.94c
78.40±0.01a
1.36
Phenylalanine
34
39.55±1.07c
41.82±0.74b
42.77±0.79b
45.89±0.40a
2.18
Tryptophan
5
1.62±0.08c
1.81±0.03b
1.66±0.04c
2.06±0.01a
0.13
Values are mean ± Standard deviation of two determinations. Means in the same row not followed by the same superscript are significantly different (p < 0.05).
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF = Fermented
Sorghum + Mango Mesocarp + Crayfish, FSPF =Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, LSD = Least Significant Difference, *Recalculated amino acid
scores from 2007 WHO/FAO/UNU report.
Table 5: Effect of Fermentation on Non-Essential Amino Acid Composition of Sorghum-Based Complementary Foods
Amino Acid
Sample
LSD0.05
(mg/g protein)
NFSMF
FSMF
NFSPF
FSPF
Histidine
18.00±0.43a
18.36±0.21a
18.34±0.80a
19.56±0.81a
1.71
Arginine
69.80±0.07b
75.36±0.16a
74.56±1.20a
75.15±0.74a
1.97
Aspartic acid
77.47±0.04c
75.62±0.71d
84.85±0.21b
88.45±0.77a
1.49
Serine
30.06±0.38d
33.67±1.20c
36.10±0.71b
38.92±0.04a
2.01
Glutamic acid
84.97±0.24c
93.28±0.23b
86.50±1.70c
97.86±0.63a
2.56
Proline
24.92±0.54c
32.67±0.76ab
31.45±1.34b
34.62±0.73a
2.49
c
c
b
a
Glycine
38.83±1.52
40.61±0.88
44.45±0.92
48.96±1.19
3.21
Alanine
29.87±0.38c
32.93±0.59ab
31.75±1.63bc
34.77±0.66a
2.63
Cystine
12.12±0.32b
11.95±1.98b
14.75±0.21ab
15.77±0.93a
3.08
Tyrosine
22.21±0.02c
23.93±0.26b
26.60±1.41a
29.79±0.71d
2.22
Values are mean ± Standard deviation of two determinations. Means in the same row not followed by the same superscript are significantly different (p < 0.05)
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF = Fermented
Sorghum + Mango Mesocarp + Crayfish, FSPF =Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, LSD = Least Significant Difference.
Table 6: Effect of Natural Fermentation on Protein Quality of Sorghum-Based Complementary Foods
Parameter
Experimental Diet
LSD0.05
NFSMF
FSMF
NFSPF
FSPF
WSF
Milk
Basal
FCR
4.33±0.02e
6.35±0.01c
5.11±0.07d
13.74±0.54b
30.15±0.25a
3.80±0.05e
-3.18±0.01f
1.54
b
d
c
e
f
a
FCE
23.10±0.11
15.65±0.14
19.58±0.29
7.29±0.29
3.33±0.04
26.34±0.38
-31.45±0.13g
0.53
PER
2.48±0.02b
1.70±0.02d
2.06±0.04c
0.81±0.03e
0.47±0.00f
2.68±0.05a
-23.83±0.16g
0.16
b
d
c
e
f
a
g
R-PER
2.31±0.02
1.58±0.002
1.92±0.04
0.75±0.03
0.44±0.00
2.50±0.05
-22.20±0.14
0.14
NPR
4.12±0.03b
3.40±0.02d
3.68±0.06c
2.74±0.04e
3.26±0.00d
4.44±0.07a
0.20±0.16f
0.16
R-NPR
3.73±0.03b
3.08±0.01d
3.33±0.05c
2.48±0.04e
2.95±0.03d
4.02±0.07a
0.18±0.13f
0.15
Values are mean ± Standard deviation of two determinations. Means in the same row not followed by the same superscript are significantly different (p < 0.05).
Key:
NFSMF= Non-Fermented Sorghum + Mango Mesocarp + Crayfish, NFSPF= Non-Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, FSMF = Fermented
Sorghum + Mango Mesocarp + Crayfish, FSPF = Fermented Sorghum + Fluted Pumpkin Leaf + Crayfish, WSF = Whole Sorghum Flour, FCR = Food Conversion
Ratio, FCE = Food Conversion Efficiency, PER = Protein Efficiency Ratio, R-PER = Relative Protein Efficiency Ratio, NPR = Net Protein Ratio, R-NPR = Relative
Net Protein Ratio.
97
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Amino Acid Composition of Sorghum-Based
Complementary Foods: The essential amino acid
composition of the blends met the amino acid pattern
by WHO/FAO/UNU (2007). The amino acid contents
of non-fermented samples were less than their
corresponding fermented samples. This could be due
to the synthesis of some essential amino acids during
solid
state
fermentation.
The
tryptophan
concentration of the blends increased significantly
(p<0.05) with fermentation. This could also be
attributed to the synthesis of some amino acids by
microorganisms during solid state fermentation
(SSF). The increase in amino acids in the fermented
product shows that the carbohydrate utilization is
closely proportional to protein production during solid
substrate fermentation. Singh et al. (1990) reported
that single cell protein produced by Aspergillus niger
contained 30.4% crude protein and had an essential
amino acid profile featuring a high lysine content and
appreciable amounts of methionine and tryptophan,
and 12.9% fat, which comprised of all the essential
fatty acids.
with fermented sorghum meal had a mean body
weight loss of -2.8g; whereas those fed diets
containing non-fermented sorghum meal had a mean
body weight gain of 13.8 g. The rats fed on FSPF and
WSF could not show any marked increase in weight
during the feeding period of 28 days.
Protein
Quality
of
Sorghum-Based
Complementary Foods: The values obtained for
feed conversion efficiency (FCE) were a reflection of
the feed conversion ratio (FCR) values since FCE is
the reciprocal of FCR expressed as a percentage.
The highest feed conversion efficiency was observed
for the reference diet (Milk) followed by
NFSMF>NFSPF>FSMF>FSPF in that order. The
FCE decreased with increase in food consumption
while the FCR increased with decrease in food
consumption during rat bioassay. FCR and FCE are
important calculations for growth which can be used
to determine if the diets are being used as efficiently
as possible (Goddard, 1996). In this study, the FCR
of 4.33 to 5.11 and FCE of 23.10 to 19.58 for NFSMF
and NFSPF respectively indicated good growth. The
lower value of FCR is indicative of the better
utilization of the diet. Protein Efficiency Ratio (PER)
indicates the relationship between weight gain in the
test animals and the corresponding protein intake.
The PER (ability of protein to support growth) of
NFSMF (2.48) compared well with the PER of milk
diet (2.68). This observation agreed with the findings
of Okoye (1992) who reported that the PER of casein
was 2.50. This observation demonstrated the positive
contribution of crayfish to the test diet. Protein
Advisory Group, PAG (1971) recommended a PER
value of 2.10 for complementary foods. Net protein
ratio (NPR) and relative NPR indices accounted for
the maintenance requirement of the weanling rat. It
relates the weight changes in the animals fed the test
diets to those fed the control diet (Shiriki et al., 2015).
The non-essential amino acids also followed the
same pattern as NFSMF had lower values compared
to FSMF in most cases. The amino acid content of
FSPF was slightly higher than NFSPF except for
aspartic acid and cystine. The reduction in amino
acids with fermentation may be due to its utilization
for growth and production of enzymes and other
organic compounds by the microorganisms during
SSF. Reduction in the concentration of some amino
acid of the blends during solid state fermentation was
in agreement with the findings of Madgi (2011) who
observed a reduction in amino acid content of pearl
millet after 24h fermentation.
Growth Response of Rats Fed with Sorghumbased Complementary Foods: The results of the
growth response of rats fed with the test diets,
NFSMF, NFSPF, FSMF and FSPF as presented in
Figure 1 revealed that NFSMF competed keenly with
the standard diet (milk) followed by NFSPF and
FSMF with FSPF recording the least growth
performance. The encouraging performance of the
non fermented diets could be partly due to absence
of flovour and taste imparted by fermentation leading
to increased consumption and subsequent increase
in weight. The high performance of NFSMF and
NFSPF may be ascribed to inclusion of crayfish
which improved the protein quality of the diets. The
low performance of the fermented diets could be due
to undesirable flavour and taste imparted by
fermentation. Nuria et al. (1984) reported that rats fed
CONCLUSION
Fermentation increased vitamin A, B12 and D as well
as the essential amino acid contents of the
blends. The inclusion of crayfish improved the
quality of the protein. Bioassay indicated that
samples, NFSMF and NFSPF exhibited good
growth behaviour, which was evident by good
PER values.
REFERENCES
Afify, A. M. R, El-Beltagi, H. S., Abd El-Salam, S. M. and
Omran, A. A. (2011a). Bioavailability of iron, zinc,
phytate and phytase activity during soaking and
98
Amer. J. Food & Nutr., 2016, 6(3): 91-100
germination of white sorghum varieties. PLoS ONE
6(10):25512, 1-7.
FAO, IFAD and WFP. (2015). The State of Food Insecurity
in
the
World
2015.
Meeting the 2015 international hunger targets: taking
stock
of
uneven
progress.
Rome, FAO.
AOAC. (2005).Official methods of analysis (18th ed.).
Arlington: Association of Official Analytical Chemists.
Bally, I. S. E (2006). Mangifera indica (mango), ver. 3.1.
In: Elevitch, C.R. (ed.). Species Profiles for Pacific
Island Agroforestry. Permanent Agriculture
Resources
(PAR),
Hōlualoa, Hawai‘i.
<http://www.traditionaltree.org>.
FAO/WHO. (1991). Protein quality evaluation. Report of
Joint FAO/WHO Expert Consultation. FAO Food and
Nutrition paper 51. FAO/WHO. Rome, Italy, pp.1–66.
Fasuyi, A. O (2006). Nutritional potentials of some tropical
vegetable leaf meals: chemical characterization and
functional properties, African Journal of Biotechnology.
5 (1): 049-053
Bentley M. E., Dickin K .L., Mebrahtu S., Kayode B., Oni
G.A., Verzosa C.C., Brown K.H., Idowu J. R (1991).
Development of a nutritionally adequate and culturally
appropriate weaning food in Kwara State, Nigeria: an
interdisciplinary approach. Soc. Sci. Med. (33): 1103–
1112.
Goddard, S (1996). Feed Management in Intensive
Aquaculture, Chapman and Hall, New York, pp: 194.
Guptill, K. S., Esrey, S. A., Oni G.A., Brown, K. H (1993).
Evaluation of a face-to-face weaning food intervention
in Kwara State, Nigeria: Knowledge, trial, and adoption
of a home-prepared weaning food. Soc. Sci. Med., 36,
665–672.
Daelmans, B. and Saadeh, R (2003) Global initiatives to
improve complementary feeding. in: SCN Newsletter:
Meeting the challenge to improve complementary
feeding. United Nations System Standing Committee
on Nutrition (ed. A. D. Moreira). Lavenhem Press, UK,
pp.10–17.
Hamdi, H. A. S (2001). Muscle and exoskeleton extracts
analysis of both fresh and marine crustaceans
Procambarus clarkii and Erugosquilla massavensis.
African Journal of Pharmacy and Pharmacology, 5(13),
pp. 1589-1597. http://www.academicjournals.org/AJPP
DOI: 10.5897/AJPP11.360
Dewey, K. G. and Brown, K. H (2003). Update on technical
issues concerning complementary feeding of young
children in developing countries and implications for
intervention programs. Food Nut Bull. 24: 5-28.
Dicko, M. H., Gruppen, H., Zouzouho, O. C., Traore, A. S.,
van Berkel, W. J. H. and Voragen, A. G. J (2006).
Effect of germination on the activities of amylases and
phenolic enzymes in sorghum varieties grouped
according to food end - use properties. Journal of The
Science of Food and Agriculture 86: 953-963.
IOM (Institute of Medicine) (2005). Dietary reference intake:
Water, potassium, sodium, chloride and sulfate.
National Academy press, Washington, DC
Kazanas, N. and Fields, M. L (1981). Nutritional
improvement of sorghum by fermentation. Journal of
Food Science. 46: 819-821.
Dina, O. A, Adedapo, A. A, Oyinloye, O. P. and Saba, A. B
(2006). Effect of Telfairia occidentalis extract on
experimentally induced anaemia in domestic rabbits.
Afr J Biomed Res., 3: 181-183.
Kazanas, N., Ray, W. E., Marion, L. F. and John, W. E
(1984). Toxic Effects of Fermented and Unfermented
Sorghum Meal Diets Naturally Contaminated with
Mycotoxinst. Applied and Environmental Microbiology,
47(5): 1118-1125
Madgi, A. O (2011). Effect of traditional fermentation
process on the nutrient and antinutrient content of
pearl millet during preparation of Lohoh. Journal of
Saudi Soociety of Agricultural Sciences. 10: 1 – 6.
Obizoba, I. C (1985). Evaluation of the protein quality of
rice supplemented with bean or crayfish in rats. Plant
Food Hum. Nutr. 35: 43 – 49.
Okoye Z. S. C (1992). Biochemical Aspects of Nutrition
Prentice-Hall of India, New Delhi pp 147-195
Mostafa, M. E., Ibrahim, S. A., Samya, S. and Ahmed, Z
(2013). Production of vitamin B12 and folic acid from
agricultural wastes using new bacterial isolates.
African Journal of Microbiology Research. 7(11):m966
– 973.
Onuorah, C. E. and Akinjede, F. A (2004). Comparative
evaluation four formulated weaning foods and a
commercial product. NIFOJ. 22: 48 – 53.
Murdock, F. A. and Fields, M. L. (1984). B-vitamin content
of natural lactic acid fermented cornmeal. Journal of
Food Science 49: 373-375.
PAG (1971). Guidelines on Protein Rich Mixtures for Use in
Weaning Foods. Protein Advisory Group, United
Nations, 45-76.
Nout, M. J. R (1993). Processed weaning foods for tropical
climates. Int. J. Food Sci. Nutri., 43: 213–221.
Pellet, P. L. and Young, V. R (1980). Nutritional evaluation
of protein foods. UN University, World hunger
programme, Food and nutrition bulletin supplement 4.
Tokyo. The United Nations University.
99
Amer. J. Food & Nutr., 2016, 6(3): 91-100
Maize, Soybean and Peanut Fortified with Moringa
oleifera Leaf Powder. Food and Nutrition Sciences, 6,
494-500. http://dx.doi.org/10.4236/fns.2015.65051
Pelto, G., Levitt, E. and Thairu, L (2003). Improving feeding
practices: current patterns, common constraints, and
the design of interventions. Food Nutr. Bull. 24: 45 –
82
Singh, K., Linden, C. J., Johnson, E. J. and Tengerdy, P. R
(1990). Bioconversion of wheat straw to animal feed by
solid substrate fermentation or ensiling. Indian J.
Microbiol., 30(2): 201-208.
Quesada-Chanto, A., Afschar, A. S., Wagner, F (1994).
Microbial production of propionic acid and vitamin B 12
using molasses or sugar, App. Microbiol. Biotechnol.
41: 378-383.
Suntornsuk, L., Gritsanapun, W., Nilkamhank, S. and
Paochom, A (2002). Quantification of vitamin C
content in herbal juice using direct titration. Journal of
Pharmaceutical and Biomedical Analysis 28: 849- 855.
Rasaco, B. A (2002). Protein Quality Tests. In: Nielson, S.
S. (Ed). Introduction to the Chemical Analysis of
Foods. CBS Publishers. New Delhi. Pp. 74 – 94.
Sengev, I. A., Ingbian, E. K and Gernah, D. I (2010).
Sensory and storage properties of Instant Kunun-zaki:
a non-alcoholic fermented sorghum beverage
supplemented with mango mesocarp flour. NIFOJ.
28(2): 336 – 348.
Wahua, T. A. T (1999). Applied Statistics for Scientific
Studies. African Link Books Ltd., Ibadan.
Shehab, G. G., Kansowa, O. A, and El-Beltagi, H. S (2010).
Effects of various chemical agents for alleviation of
drought stress in rice plants (Oryza sativa L.). Not Bot
Horti Agrobo 38(1):139-148.
World
Health
Organization/Food
and
Agriculture
Organization/United Nations University (2007). Protein
and Amino Acid Requirements in Human Nutrition
Report
of
a
Joint
WHO/FAO/UNU Expert Consultation. WHO Technical
Report
Series
no.
935.
Geneva:
WHO.
Walker A.F (1990). The contribution of weaning foods to
protein-energy malnutrition. Nutr. Res. Rev., 3: 25–47.
Shiriki, D., Igyor, M. A. and Gernah, D. I (2015). Nutritional
Evaluation of Complementary Food Formulations from
100