Microbiological and Proximate Analysis of Okpehe, a Locally Fermented Condiment
Patience
Temitope Fowoyo*
Department of Biosciences, Salem University, Lokoja, Nigeria
*Corresponding author: Patience Temitope Fowoyo, Department of Biosciences, Salem University, Lokoja, Nigeria. Tel: +2347068037034; Email: patbello83@gmail.com
Received Date: 02 October, 2017; Accepted Date: 20 October, 2017; Published Date: 27 October, 2017
Citation: Fowoyo PT (2017) Microbiological and Proximate Analysis of Okpehe, a Locally Fermented Condiment. Food Nutr J 2: 153. DOI: 10.29011/2575-7091.100053
1. Abstract
1. Introduction
·
Determination of moisture content: The method
according to James (1995)[15] was employed.
·
Determination of crude lipid: Extraction of the fat and
quantification was carried out using soxhlet apparatus as adopted by AOAC
(2002) [16]using petroleum ether as solvent.
·
Determination of crude fibre content: The method according to AOAC
(2002) [16]was employed.
·
Determination of total ash content: The method according to AOAC (2002) [16] was
employed.
·
Determination of crude protein: The
Micro-Kjeldhal method adopted by AOAC (2002) [16] was employed.
·
Determination of carbohydrate content of okpehe
samples:
The method according to AOAC (2002) [16] was employed. The carbohydrate or nitrogen free extract
content of the samples was determined by the difference obtained by subtracting
the sum of percentage ash, crude protein, crude fiber and ether extract from
100% on dry matter basis.
% Carbohydrate = 100 - (% ash
+ % crudeprotein + % etherextract + % crude fibre)
The method according to Eby (2014) [20]was employed for determination of vitamin A, C and E content.
3.1. Microbiological quality of okpehe samples
Bacillusspecies was the
most predominant in the okpehe samples, followed closely by S. aureus
and then E.
coli, which is the
least frequently occurring organism as shown in Table 6.
Total fungal
counts revealed higher counts of A. niger in samples from garage market -Anyigba
and new market -Lokoja, while Mucor and Paecilomyces species occurred in frequently
between samples from garage market- Anyigba and new and old markets, Lokoja. Aspergillus niger
was the highest occurring fungi followed by Mucor sp. and Paecilomyces sp. occurring in relatively irregular patterns as
shown in Table 7.
3.2. Nutritional composition of okpehe samples
Alkaloids,
steroids, flavonoids, saponins, resins and phenols were found to be present
while anthraquinones, triterpenoids, tannins and oxalate were found to be
absent from all samples analyzed(Table 9).
4. Discussion
The
higher bacterial load obtained in sample D (okpehepurchased from Unity square, Anyigba) may be attributed to
the production procedure carried out for the sample along with the location of
display and sale, which is a mini-market which also doubles as a motor-park
garage, and a general meeting point for community functions, further
emphasizing the need for appropriate health care and conscientious contact
control precautions.
Sample |
Sampling location |
Colour |
Smell |
Texture |
A |
Ojogobi, Ankpa |
Black |
Strong ammoniacal smell |
Soft |
B |
Otukpo, Ankpa |
Brown |
Strong ammoniacal smell |
Soft |
C |
Garage market |
Black |
Strong ammoniacal smell |
Soft |
D |
Unity square, Anyigba |
Black |
Strong ammoniacal smell |
Soft |
E |
Ejah, Idah |
Brown |
Strong ammoniacal smell |
Soft |
F |
Inikpi, Idah |
Brown |
Strong ammoniacal smell |
Soft |
G |
Old market, Lokoja |
Light brown |
Mild ammoniacal smell |
Hard |
H |
New market, Lokoja |
Light Brown |
Mild ammoniacal smell |
Hard |
Table 1: Physical characteristics of okpehesamples.
Sample |
Sampling location |
pH |
Titratable acidity (g/L) |
A |
Ojogobi, Ankpa |
7.46 |
0.38 |
B |
Otukpo, Ankpa |
7.69 |
0.35 |
C |
Garage market |
7.83 |
0.27 |
D |
Unity square, Anyigba |
7.79 |
0.30 |
E |
Ejah, Idah |
7.15 |
0.43 |
F |
Inikpi, Idah |
7.34 |
0.40 |
G |
Old market, Lokoja |
6.72 |
0.50 |
H |
New market, Lokoja |
6.91 |
0.45 |
Table 2: Physicochemical parameters of okpehesamples.
Samples |
Sampling location |
Total bacterial count (cfu/g) |
Total fungal count (cfu/g) |
A |
Ojogobi, Ankpa |
1.7× 106 |
1.0× 104 |
B |
Otukpo, Ankpa |
1.5× 106 |
1.0× 104 |
C |
Garage market |
1.6× 106 |
3.0× 104 |
D |
Unity square, Anyigba |
1.8× 106 |
1.0× 104 |
E |
Ejah, Idah |
1.2× 106 |
2.0× 104 |
F |
Inikpi, Idah |
1.4× 106 |
1.0× 104 |
G |
Old market, Lokoja |
9.5× 105 |
3.0× 104 |
H |
New market, Lokoja |
1.1× 106 |
4.0× 104 |
Table 3: Total fungal and bacterial count of okpehe samples.
Isolates Code |
Cultural Characteristics |
Gram Reaction |
Spore Characteristics |
Starch Hydrolysis |
Motility |
Citrate |
Urease |
Catalase |
Oxidase |
Indole |
Sugar Fermentation |
Probable Organism |
||||
Glucose |
Fructose |
Maltose |
Sucrose |
Lactose |
||||||||||||
Aoj1, Lom3, Lnm1, Iim1, Aot4, Aus2, Ijm3 |
Small and mucoid, circular, slightly raised, entire, opaque, creamy, dry. |
Gram +ve rods |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
A |
A |
A |
A |
A |
Bacillus subtilis |
Aoj2, Aot2, Aus3, Agm2, Iim4, Lom2, Lnm2 |
Large, irregular, flat, lobate, opaque, whitish, smooth. |
Gram +ve rods in clusters |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
A |
AG |
A |
A |
A |
Bacillus licheniformis |
Aoj3, Agm1, Aot5 |
Medium sized, circular, raised, translucent, light metallic green, smooth. |
Gram -ve rods |
- |
- |
- |
- |
+ |
+ |
+ |
AG |
AG |
AG |
A |
AG |
Escherichia coli |
|
Aoj4, Aot3, Aus1, Agm3, Ijm4, Ijm3, Lom1. Lnm3 |
Moderate, undulate (wavy), slightly raised, convex, opaque white, dry and slightly wrinkled. |
Gram +ve rods |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
A |
A |
A |
A |
A |
Bacillus megaterium |
Aoj5, Aot1, Aus4, Ijm2, Iim2, Lom4, Lnm4 |
Moderate, Circular, Convex, Entire (smooth), Opaque, Pale yellow, glistening. |
Gram +ve coccus in clusters |
- |
- |
+ |
- |
- |
+ |
- |
+ |
AG |
A |
A |
A |
A |
Staphylococcus aureus |
Table 4: Morphological and biochemical characteristics of bacterial isolates from okpehesamples.
Isolate code |
Colour |
Texture |
Shape |
Type of hyphae |
Conidiophore |
Sporangium |
Collumella |
Probable Organism |
Agm1,Lom1, Lnm2 |
Black |
Wrinkled |
Round |
non-septate |
Non-branched |
- |
- |
Aspergillus niger |
Agm2, Ijm1, Lom2, Lnm1 |
Cotton white |
Rough and loose |
Rhizoid |
non-septate |
Non-branched |
+ |
Well developed |
Mucorsp. |
Aus1, Ijm2, Lom3, Lnm3 |
Greenish brown |
Smooth |
Irregular |
non-septate |
Branched |
- |
- |
Paecilomycessp. |
Table 5: Macroscopic and microscopic characteristics of fungal isolates from okpehesamples.
Sample |
B. subtilis(cfu/g) |
B. licheniformis(cfu/g) |
B. megaterium(cfu/g) |
E. coli (cfu/g) |
S. aureus (cfu/g) |
A |
4.7 × 105 |
3.5 × 105 |
3.7× 105 |
2.1× 105 |
3.0× 105 |
B |
4.3× 105 |
3.6× 105 |
4.1× 105 |
1.6× 105 |
1.9× 105 |
C |
5.1× 105 |
4.4× 105 |
4.7× 105 |
1.0× 104 |
2.0× 105 |
D |
5.3× 105 |
4.3× 105 |
3.9× 105 |
2.4× 105 |
3.0× 105 |
E |
4.7× 105 |
2.5× 105 |
2.6× 105 |
1.0× 104 |
2.7× 105 |
F |
5.4× 105 |
3.3× 105 |
3.9× 105 |
1.0× 104 |
2.0× 105 |
G |
3.8× 105 |
2.3× 105 |
1.9× 105 |
1.0× 104 |
1.5× 105 |
H |
4.3× 105 |
3.2× 105 |
2.9× 105 |
1.0× 104 |
1.5× 105 |
Table 6: Total count of bacterial species in okpehe samples.
Sample |
Aspergillus niger(cfu/g) |
Mucor sp. (cfu/g) |
Paecilomyces sp.(cfu/g) |
A |
- |
- |
- |
B |
- |
- |
- |
C |
2 × 104 |
1 × 104 |
- |
D |
1 × 104 |
- |
- |
E |
1 × 104 |
- |
1 × 104 |
F |
- |
- |
- |
G |
1 × 104 |
1× 104 |
1× 104 |
H |
2× 104 |
1 × 104 |
1 × 104 |
Table 7: Total count of fungal species in okpehesamples.
Sample |
Proximate Composition (% per g of sample) |
|||||
Moisture Content |
Ash Content |
Lipid Content |
Fibre Content |
Protein Content |
Carbohydrate Content |
|
A |
7.92 ± 0.10b |
5.85 ± 0.12b |
11.32 ± 0.32a |
4.67 ± 0.20a |
34.23 ± 0.01d |
47.06 ± 0.01c |
B |
8.01 ± 0.04a |
5.72 ± 0.83b |
10.67 ± 0.01b |
4.52 ± 0.10a |
36.42 ± 0.02c |
46.45 ± 0.24d |
C |
8.64 ± 0.05a |
5.31 ± 0.05b |
9.23 ± 0.05c |
4.23 ± 0.11a |
40.28 ± 0.33a |
41.56 ± 0.12f |
D |
8.36 ± 0.13a |
5.65 ± 0.05b |
11.45 ± 0.01a |
4.86 ± 0.25a |
38.34 ± 0.15b |
44.32 ± 0.30e |
E |
6.97 ± 0.03c |
6.23 ± 0.02a |
8.13 ± 0.01d |
3.24 ±0.04b |
31.76 ± 0.2f |
47.54 ± 0.14c |
F |
7.41 ± 0.03b |
5.92 ± 0.05b |
10.34 ± 0.01b |
3.82 ± 0.20b |
32.72 ±0.01e |
47.62 ± 0.22c |
G |
4.74 ± 0.02e |
6.09 ± 0.83a |
7.34 ± 0.05e |
3.58 ± 0.18b |
32.53 ± 0.01e |
51.63 ± 0.18a |
H |
5.13 ± 0.24d |
5.85 ± 0.11b |
9.53 ± 0.05c |
3.74 ± 0.01b |
32.84 ± 0.22e |
49.21 ± 0.21b |
*Values are means of triplicate determination ± SD) Means in the same column with different superscripts are significantly different at (p < 0.05) |
Table 8: Proximate composition of okpehe samples.
Sample |
Anti-nutritional Factors |
|||||||||
Alkaloids |
Steroids |
Flavoniods |
Saponins |
Resins |
Anthraquinones |
Phenols |
Triterpeniods |
Tannins |
Oxalate |
|
A |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
B |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
C |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
D |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
E |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
F |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
G |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
H |
+ |
+ |
+ |
+ |
+ |
- |
+ |
- |
- |
- |
Table 9: Qualitative anti-nutritional screening of okpehe samples.
Sample |
Phytochemical content (%) |
|||||
|
Alkaloids |
Steroids |
Flavonoids |
Saponin |
Resins |
Phenols |
A |
0.31 |
0.22 |
0.21 |
1.10 |
0.46 |
0.38 |
B |
0.28 |
0.31 |
0.36 |
0.94 |
0.38 |
0.64 |
C |
0.22 |
0.34 |
0.23 |
0.84 |
0.75 |
0.52 |
D |
0.39 |
0.25 |
0.20 |
0.91 |
0.88 |
0.34 |
E |
0.36 |
0.27 |
0.32 |
1.01 |
0.48 |
0.28 |
F |
0.39 |
0.25 |
0.18 |
0.88 |
0.41 |
0.32 |
G |
0.26 |
0.33 |
0.26 |
0.72 |
0.64 |
0.38 |
H |
0.21 |
0.42 |
0.28 |
0.93 |
0.87 |
0.46 |
Table 10: Quantitative ant nutritional screening of okpehesamples.
Sample |
Mineral (mg/g) |
|||||||
Potassium |
Cadmium |
Copper |
Calcium |
Iron |
Manganese |
Zinc |
Lead |
|
A |
2.295 |
BDL |
0.092 |
0.556 |
0.105 |
0.061 |
0.142 |
BDL |
B |
2.951 |
BDL |
0.087 |
0.411 |
0.103 |
0.065 |
0.147 |
BDL |
C |
2.101 |
BDL |
0.083 |
0.504 |
0.114 |
0.073 |
0.162 |
BDL |
D |
3.111 |
BDL |
0.088 |
0.523 |
0.108 |
0.078 |
0.158 |
BDL |
E |
2.745 |
BDL |
0.094 |
0.492 |
0.121 |
0.071 |
0.162 |
BDL |
F |
3.015 |
BDL |
0.096 |
0.516 |
0.126 |
0.089 |
0.155 |
BDL |
G |
3.496 |
BDL |
0.073 |
0.447 |
0.106 |
0.087 |
0.172 |
BDL |
H |
2.166 |
BDL |
0.08 |
0.572 |
0.104 |
0.088 |
0.144 |
BDL |
BDL- below detection limits |
Table 11: Mineral composition of okpehe samples.
Sample |
Vitamin A (%) |
Vitamin E (mg/ml) |
Vitamin C (mg/ml) |
A |
0.006 |
0.002 |
BDL |
B |
0.002 |
0.001 |
BDL |
C |
0.004 |
0.002 |
BDL |
D |
0.001 |
0.002 |
BDL |
E |
0.003 |
0.001 |
BDL |
F |
0.005 |
0.002 |
BDL |
G |
0.003 |
0.001 |
BDL |
H |
0.001 |
0.001 |
BDL |
Table 12: Vitamin content of okpehe samples.
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