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This study enumerates the bacterial and fungal isolates responsible for the fermentation of millet
sourdoughs, evaluates the antimicrobial safety of the sourdough starters and the quality of bread
produced thereafter. The individual flours were spontaneously fermented in the ratio 1:1 (w/v) for
48 h. At the end of the fermentation, the microbial consortium of the sourdough meals obtained
were determined and also screened for antagonistic activity against selected pathogens. All the
sourdough starters revealed strong clearance zones against the selected food borne pathogens
with values ranging from 5.00 to 16.1 mm. The proximate compositions of the breads were
improved by the sourdough fermentation process compared with the raw flour. The moisture
content of flour determined ranged from 7.48 to 11.20% while the protein, fat, ash, crude fibre and
carbohydrate were 9.53 to 12.4%; 2.13 to 2.78%; 0.82 to 2.87%; 2.10 to 4.67% and 71.83 to 76.33%
respectively. The carbohydrate content of the breads reduced with fermentation and ranged from
63.10% to 73.33% while the protein, ash, crude fibre and fat content increased and ranged from
11.57 to 15.31%; 2.39 to 3.08%; 1.76 to 3.62% and 7.15 to 12.32% respectively. The highest value
for calcium 48.86 mg/100 g was exhibited by pearl millet bread. Sensory evaluation showed
significant difference (P < 0.05) in the appearance, taste, colour, aroma and texture of the bread
samples. The findings have suggested that indigenous wholegrain cereals could be employed for
bread production, which has the potential to improve the nutritional profile and safety of cereal
fermented foods which forms the bulk of the diet of rural dwellers in sub-Saharan Africa.
INTRODUCTION
Consumers’ interest in health- promoting, nutritious, are present in the bran such as oligosaccharides,
functional and whole grain rich foods is significantly phytochemicals and dietary fibre (Slavin, 2010).
increasing, particularly against several western chronic Consumption of rich fibre foods have long been proven to
diseases such as diabetes, celiac disease and other provide health benefits. (Jideani, 2012) Utilizing the
cardiovascular diseases (Pereira et al., 2002; Coda et al., sourdough technique in the development of fibre rich
2012). White pan bread and other bakery products functional foods such as wholegrain sourdough bread,
produced from the endosperm flour represents 83% of the cake and other baked goods is an emerging technology in
kernel, characterized by a high soft elastic crumb structure sub Saharan Africa and could be suitable in enhancing the
and high volume owing to the development of gluten technological and nutritional properties of indigenous
network which entraps the gas produced from the
fermentation by yeast and adds to the unique cellular *Corresponding Author: ADISA, Abosede Margaret;
structure. Nutritionally, white pan bread is inferior to the Department of Food Science and Technology, Joseph Ayo
wholegrain which comprise germ 3% and bran 12% (Sidhu Babalola University. Ikeji Arakeji. P. M. B. 5006, Ilesha,
et al., 2007; Edema and Sanni, 2008). Healthy metabolites Osun State, Nigeria. E-mail: abosedeadis@gmail.com
The raw materials used are pearl millet (Pennisetum This was done using the agar well diffusion method. The
glaucoma), White fonio (Digitaria exilis), Black fonio test organisms were grown in nutrient broth at 37 oC for 24
(Digitaria iburua) and finger millet (Eleusine coracana). h. The inoculum was standardized to give a cell density of
They were all purchased from Owena market, Ilesa, 108 cfu ml-1 approximately correseponding to 2 McFarland.
Nigeria. Pearl and finger millet were sorted differently to 100 µl of the test inoculum was spread evenly by
remove stones and debris, while the fonio grains were swabbing on Muellar-Hinton agar plates and 3 wells each
winnowed separately to remove extraneous materials. All of 6 mm diameter were bored into the plates with the aid of
materials were ground into fine flour using the attrition disc a sterile cork-borer. The wells were then filled with 100 µl
mill, packaged in air tight plastic containers and stored in aliquot of freshly prepared sourdough (1;1 w/v) in a laminar
the refrigerator (4 oC ) until required. All chemicals used airflow cabinet in order to maintain sterile condition. The
were of analytical grade. inoculated plates were incubated at 37 oC for 24 h and the
significant difference (p<0.05) in the crude fibre content of requirement set by FAO/WHO of 1 × 106 cfu/g of live
the fonio grains WIF and BAF which were 0.82% and organisms (Table 3). The highest number of Lactic acid
0.85% respectively. Carbohydrate contributes to the bulk bacteria was from PEM with count of 4.9 × 106 cfu/g and
of the energy intake of human diet. High value of the lowest was found in FIM with a count of 2.0 × 106 cfu/g,
carbohydrate content was observed for all the flours and going by this initial count, wholegrain millet sourdoughs
corresponds to the values obtained by other authors (Coda can be considered as good probiotic sources.
et al., 2010; Jideani, 2012; Ballogou et al., 2013). The Ash
content of the flours ranged between 2.13% and 2.78% Table 3. Microbial counts cfu g100- g of LAB and yeast
and corresponds with the result of Serna (2003) for higher Sourdoughs LAB (×106) Yeast (×105)
values for ash content which is an indication of rough PEM 4.9a 3.0b
estimate of the mineral contents of the flours . The crude FIM 2.0a 7.4ab
protein content was observed to be very high in FIM with a BAF 3.9a 9.4a
value of 12.4% followed by PEM, WIF and BAF with 10.8%, WIF 4.4a 8.5a
10.0% and 9.53% respectively. Means with different superscripts on the same column are
significantly different (p<0.05).
All the parameters analysed showed that sourdough Values are means ± SD of triplicate measurement.
fermentation enhanced the proximate composition of the PEM=pearl millet, FIM= Finger millet, WIF= White fonio,
breads (Table 2) as there were significant improvements in BAF= Black fonio
the values obtained for the sourdough bread when
compared with the raw flour. Also, the co-interaction between LAB and yeast is common
in many food fermentation matrixes, wherein Lactic acid
Table 2. Proximate composition (g 100 g-1 DM) sourdough bacteria creates the acidic environment for yeast growth,
breads and yeast in return provides vitamins and other growth
Sample Moisture Crude Total Crude Fat Carbohydrate factors. In addition, LAB is represented in fermented foods
content protein ash fibre because they are able to survive high acidic conditions and
PEM 5.83** 12.9** 2.39** 3.44** 12.3** 63.1** ±0.01 also have the ability to provide a high level of lactic acid
±0.01 ±0.0 ±0.01 ±0.01 ±0.02 (Wakil and Daodu, 2011).The results of the morphological
FIM 4.23** 15.3** 3.08** 3.62** 7.49** 66.3** ±0.01 and biochemical tests used to characterize the isolates
±0.01 ±0.02 ±0.02 ±0.03 ±0.01 obtained were similar to those reported by other authors
WIF 15.5** 12.2** 2.66** 1.76** 7.15** 60.7** ±0.02 (Ahmed and Kanwal, 2004; Nwachukwu et al., 2010; Goyal
±0.03 ±0.01 ±0.05 ±0.04 ±0.01 et al., 2012) and were also referenced to Bergey’s manual
BAF 9.30** 11.6** 2.55** 1.82** 7.43** 73.3** ±0.01 of modern bacteriology.
±0.03 ±0.01 ±0.01 ±0.01 ±0.02
**Means that are significantly different on the same column All the isolates identified on MRS agar were Gram positive,
(p<0.05). rod shaped, catalase negative and non-spore forming
Values are means ± SD of triplicate measurement. which are characteristic of LAB (Table 4). The result of the
PEM=pearl millet, FIM= Finger millet, WIF= White fonio, biochemical tests showed that the isolates belonged to
BAF= Black fonio, DM= Dry matter genus of lactobacillus. The presence of fungi was
investigated on malt extract agar and Saccharomyces
Sourdough is effective for the production of bread with cerevisiae was observed to be the predominant species in
increased level of flavour compounds and ultimately three of the samples, while Candida was found dominating
increasing consumers’ satisfaction. Generally, there was in pearl millet sourdoughs as presented in (Table 5). This
steady decrease in carbohydrate content of all the bread is attributed to the variation in the utilization of sugar
samples compared with their respective flour with values among the yeast. The ability of the yeast to assimilate
ranging from 63.1%-73.3% to 71.8-76.3%. Highest crude glucose, galactose, sucrose and raffinose confirmed for S.
fat content of 10.8% was observed for PEM bread, while cerevisiae, which is similar to the findings of Al-Otaibi.
there was no significant difference (p<0.05) observed for 2012. The presence of yeast in these sourdoughs relates
FIM, WIF and BAF. After baking FIM bread had the highest well with the identification and characterization of yeast
value for crude protein and crude fibre (13.9% and 3.49%) from other African fermented foods. In Ghanaian pito,
respectively. This corresponds with the values reported by Demuyakor and Ohta (1991) reported a high
Rehman et al. (2006) and Cruz et al. (2011). predominance of Saccharomyces cerevisiae (33%) and
presence of Candida spp. (17%) amongst other genera.
Microbial count and distribution of isolates during While investigating Nigerian sorghum beer, Sanni (1993)
sourdough fermentation observed occurrence of several species, Candida spp.,
Saccharomyces cerevisiae, Geotrichum candidum,
Yeast and LAB species dominated the fermentation Torulaspora delbrueckii to be dependent on the type of
process and the high population obtained showed that all beer.
samples of wholegrain millet sourdoughs met the minimum
Catalase test - - - -
Motility - - - -
Lactose + + + +
Glucose + + + +
Sucrose + V + +
Raffinose - + V +
Spore forming - - - -
CO2 - - - -
Growth in 10% NaCl + + + +
Presumptive org Lb. fermentum Lb. plantarum Lb. plantarum Lb. plantarum
+=positive, - =negative, v =variable, +G= gram positive, (R)= rod shape
PEM=Pearl millet, FIM = Finger millet, BAF = Black fonio, WIF= White fonio
Glucose + + + +
Galactose ̶ + + +
Lactose ̶ ̶ ̶ ̶
Melibiose ̶ ̶ ̶ ̶
Sucrose + + + +
Raffinose ̶ ̶ ̶ +
Spore forming ̶ ̶ ̶ ̶
Urea hydrolysis ̶ + + +
Resistance to ̶ ̶ ̶ ̶
Cycloheximide
Growth in 10% NaCl V ND ND V
Growth at 37 oC + + + +
Growth at 42 oC ̶ + + +
Presumptive Candida sp. Saccharomyces Saccharomyces Saccharomyces
organisms cerevisiae cerevisiae cerevisiae
positive, - negative, v variable, ND not determined
PEM=Pearl millet, FIM = Finger millet, BAF = Black fonio, WIF= White fonio.
Antagonistic activity against selected pathogens was obtained from brown fonio sourdough (SBF) which
gave an inhibition of 3mm diameter against Klebsiella sp.
All the sourdoughs displayed large clear zones of Pearl millet sourdough showed lowest activity against all
inhibitions to varying degrees against all the tested tested pathogens. The result is presented in Fig. I. The
pathogens, thus confirming the antimicrobial potential of relatively large inhibition zone is an indication that fresh
the sourdoughs. The highest inhibition zone was observed sourdoughs oppose bioactive components such as
by finger millet sourdough (SFM) which gave a clearance bacteriocins which are antimicrobial peptide possessing
diameter of 16.1 mm against Pseudomonas sp, this was bacteriostatic or bactericidal activity against the selected
closely followed by E. coli with 15.0 mm. the lowest activity test pathogens.
influence of fermentation on the sourdough. Food acceptability of the breads. The breads had a characteristic
fermentation offers numerous benefits which include pleasant aroma and sour taste which is unique to
improvement of nutritional qualities of protein and sourdough bread fermented with heterofermentative Lactic
carbohydrate digestibility (Sanni, 1993, Onilude et al., acid bacteria (Banu et al., 2001). White fonio bread
1999). recorded the highest overall acceptability probably due to
panellists’ preference for colour and taste. The dark brown
Physical properties of the sourdough breads colour of finger millet bread, medium cracks and hardness
of the bread could be responsible for the low scores in its
The physical properties of the sourdough breads are overall acceptance.
presented in (Table 7). There was significant difference
(p<0.05) in the breads with regards to the loaf specific Shelf life of bread
volume (LSV) and the values ranged from 1.75 cm3 to 3.66
cm3. Finger millet bread had the highest weight of 141.87 After four days storage at room temperature the breads
g while brown fonio bread has the lowest weight of 123.0 showed a little growth of mould. This observation can be
g. linked to the use of sourdough in the preparation of bread
as it has a positive effect on mould- free shelf life (Corsetti
Table 7. Physical Properties of Wholegrain Millet et al., 2003) and also due to the exopolysaccharides
Sourdough bread produced by LAB which enhanced water retention and
Height (cm) Weight (g) LV(g/cm3) LSV (cm3) functioned as hydrocolloids (Yann and Pauline, 2014)
FIM 5.73** 135.27** 500.61** 3.66**
±0.15 ±1.23 ± 16.2 ± 0.15
PEM 4.66** 141.87** 249.1** 1.75** CONCLUSION
±0.10 ± 0.58 ± 8.67 ± 0.06
BAF 4.66** 123.03** 447.77** 3.59** From this present study it was observed that sourdough
±0.10 ± 0.61 ± 0.70 ± 0.15 fermentation improved the proximate composition and
WIF 5.23** 126.5** 447.77** 3.50a mineral bio-availability of the bread samples significantly
±0.58 ± 2.72 ± 0.70 ± 0.08 (p<0.05). The highest improvement was observed in
**Means that are significantly different on the same column sample FIM. The potential of the sourdoughs to inhibit the
(p<0.05). growth of selected pathogens is an affirmation of the
Values are means ± SD of triplicate measurement probiotic health benefit that could be derived from the
FIM = Finger millet bread, PEM = Pearl millet bread, BAF consumption of such specialty foods. Thus, the sourdough
= Black fonio bread, WIF = White fonio bread, technique when applied to indigenous substrates such as
LV = Loaf volume, LSV = Loaf specific volume. millet can provide consumers especially those at the
grassroot level with a safe, wholesome, nutritious food with
All the breads had high loaf volume which could be due to health promoting benefits that could positively ameliorate
the high retention of gas due to increased level of LAB the lives of rural dwellers in sub-Saharan regions.
activity during sourdough fermentation. This also agrees
with the findings of Crowley et al. (2002) and Tamani et al.
Conflict of Interest
(2013) which stated that sourdough fermentation
enhanced bread volume
The author(s) did not declare any conflict of interest.
Table 8. Sensory properties of the sourdough bread
Authors’ Contributions
Sensory qualities FIM PEM BAF WIF
Appearance 5.63d 6.41c 7.15a 7.36a Abosede Adisa; Designed and performed experiments,
Taste 5.22d 6.77c 7.13b 7.72a analysed data and co-wrote the paper, Ifesan Beatrice;
Colour 5.78d 6.28c 7.15b 7.52a Supervised the research, Adepeju Adefisola and
Aroma 5.66c 6.18b 6.80a 6.85a Enujiugha Victor; Co- wrote and corrected the manuscript.
Texture 5.35c 6.71b 7.42a 7.60a
Overall acceptability 5.22 c 6.37b 7.13ab 7.56a
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