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Volume 8, Issue 10, October – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
IJISRT23OCT1320 www.ijisrt.com 1376
Evaluation of the Microbial, Proximate, and Sensory
Properties of Biscuits Fortified with
Pleurotus ostreatus at Various Inclusion Levels
Rosemary A. Ajayi1* and Victor O. Oyetayo2
Department of Microbiology, School of Life Sciences, Federal University of Technology, Akure, Nigeria
Corresponding Author: Rosemary A. Ajayi1*
Abstract:- To satisfy consumer desires for more health
advantages, food products must be fortified. The
sensory, proximate, and microbiological characteristics
of biscuits enriched with Pleutotus ostreatus at different
inclusion levels were assessed. The manufacturing of
wheat flour, baking of mushroom powder biscuits
(MPB), microbiological examination of MPB, proximate
analysis, and sensory evaluation of MPB were all done
using standard procedures. The fortified mushroom
biscuit had varying mushroom compositions, ranging
from 50-100% (sample A-F). The results showed that the
composition with 90% mushrooms had the highest
bacteria count (62 CFU/ml) while 50% had 1 CFU/ml.
Various microorganisms were tentatively identified from
MPB, including Staphylococcus warnei, Bacillus niacin,
Kluyuera georgiana, B. circulans, B. licheniforms, B.
megaterium, B. barbaricue, B. cereus, B. simplex,
Aspergillus flavus, A. niger, and P. ostreatus. According
to the study, the nutritious value of wheat cookies
supplemented with mushrooms rose as mushroom
powder concentration rose. The value of the
carbohydrates was the highest at 73%, while the value of
the ash content was the lowest at 2%. Sample D, which
contained 50% each of flour and mushrooms, had the
highest crude fibre level. Higher mushroom powder
concentrations reduced the fortified biscuit's overall
acceptability (OA). Sample E (10% mushroom) had the
highest OA, scoring 9.30 0.21 for both texture and
flavour. Based on the findings, it is recommended that
the commercial production of biscuits be carried out at a
50% level of mushroom inclusion for the best
nutritional, essential mineral and sensory efficacy.
Keywords:- Mushroom, Biscuit, Proximate, Sensory,
Nutritional, Mineral.
I. INTRODUCTION
The most popular sort of snack food among bakery
goods, biscuits are created from simple, affordable, and
easily accessible raw materials. Because of their flavour and
lengthy shelf life due to their low water activity, they are
frequently consumed (Caleja et al., 2017). They are among
the most well-known bakery goods in Nigeria (Bello et al.,
2017). They are healthy snacks produced from dough that is
not particularly appetising but is transformed into a pleasant
treat by baking it (Kure et al., 1998). The main components
of biscuit dough are water, sugar, oil, and soft wheat flour.
To create a dough with a strong gluten network, they are
combined with a few other ingredients (including baking
powder, skim milk, emulsifier, and sodium metabisulphite)
(Blanco et al., 2016). The need to fortify food products like
biscuits has increased due to the increased global demand
for more functional agricultural and innovative food
products, especially as biscuits are one of the most popular
snack meals (Liyanage and Hettiarachchi, 2011).
In order to meet consumer requests for additional
advantages, food fortification plays a significant role in
boosting the amount of functional components that promote
health in bakery products (Wieca et al., 2017; Adeboboye et
al., 2020). When plant-based, macrofungal, and other
protein supplies are used with wheat flour, the protein
concentration of baked foods is greatly boosted (Ugwuona
and Obeta, 2016). Some macrofungi, such as mushrooms,
include high-quality digestible protein (10–40%),
carbohydrates (3-21%), and dietary fibre (3–35%),
depending on the species (Mallavadhani et al., 2006).
Because they are rich in proteins, vitamins, minerals, chitin,
and vital amino acids and contain few calories and fat,
mushrooms are consumed as a source of nutrition around the
world. In addition to protein and dietary fibre, mushrooms
also contain vitamins, minerals like potassium, phosphorus,
and iron, as well as carbs and carbohydrates (Valverde et al.,
2015). Additionally, mushrooms contain vitamins including
vitamin C and the B-vitamin family (thiamin, riboflavin,
biotin, niacin, pyridoxine, and panthotenic acid), as well as
vital amino acids like lysine (Adedayo, 2011; Okafor et al.,
2012).
Pleurotus ostreatus is a tasty mushroom with a low fat
and carbohydrate content, a variety of proteins, and minerals
(Ca, P, Fe, Mg), making it a superior nutritional meal (Silva
et al., 2002). After Agaricus bisporus. P. ostreatus is the
second most popular edible mushroom for cultivation and
has a substantial economic impact (Sanches, 2010). It has
been demonstrated that some grown mushrooms produce
less and grow more slowly than this species. P. ostreatus
has witnessed a large increase in international cultivation in
recent decades because of its notable tolerance of a range of
agro-climatic conditions. (Sanches, 2010; Kholoud et al.,
2014). Pleurotus ostreatus has a content of nutrients that is
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readily digestible proteins, mineral salts, vitamins, and
compounds with potent pharmacological activities, e.g.,
lovastatin and pleuran, P. ostreatus is an important
mushroom species of dietary and medical significance
(Anandhi et al., 2013; Muszyńska et al., 2014; Caz et al.,
2015).
Biscuits are often produced using wheat flour and fat,
which might be unhealthy if consumed frequently,
especially in excess, according to Caleja et al. (2017). A
class of microbial agents used to fortify food and food
products is represented by mushrooms. They have thus been
employed as good fortificants because they have been
shown to have high levels of protein, dietary fibres,
vitamins, minerals (such as potassium, phosphorus, and
iron), carbs, chitin, and vital amino acids, as well as low
levels of fat and calories (Valverde et al., 2015). The
purpose of the study is to isolate and identify microorganism
from biscuit fortified with mushroom (P. ostreatus), fortify
biscuit with mushroom (P. ostreatus) at various levels of
inclusion, and assess the proximate and sensory composition
of the fortified biscuit. Therefore, the purpose of this study
is to assess the microbiological, proximate, and sensory
qualities of biscuits enhanced with mushroom (P. ostreatus)
at different levels of inclusion.
II. METHODS
A. Cultivation of Mushroom
Tissue Culturing
PDA was produced in accordance with the
manufacturer's instructions. A 500 ml conical flask was
filled with precisely 5g of potato dextrose agar, 100ml of
water, and the conical was gently shaken to help the agar
dissolve. A combination of sterile cotton wool and
aluminium foil were used to cap the conical flask. The agar
was sterilized at 121°C for 60 minutes and allowed to cool
to 45°C pour plating. Pour plating was done aseptically into
Petri dishes. The agar was allowed to completely fill the
bottom of the Petri dish, immediately covered and kept
aside. After solidification, tissue of each species of
Pleurotus was aseptically cultured into the different Petri
dishes. Briefly, the basidiocarp of the fruiting body of a
Pleurotus species was sanitized using absolute ethanol,
longitudinally cut into two halves and tissue bits from collar
regions was picked using forceps and transferred to pre
sterilized Potato Dextrose Agar (PDA). The Petri plates
were kept in an incubator for a week at a temperature of
25°C plus 2°C. To create pure cultures, mycelium from
developing edges was carefully transferred to PDA slants
and cultured for two to three weeks (Hsu et al., 2018).
Preparation of Grain Spawn
The millet grains for spawning were thoroughly
washed and soaked for 24hours in water, and then sieved.
After this, the grains were filled halfway (to create air space
for welling and shaking) into heat resistant. The autoclave
was used to sterilise the bottles for 15 minutes at 121°C.
After chilling, the grains in the bottles were inoculated with
6 cm mycelial discs of the desired Pleutorus species and
cultured for roughly 2 weeks at 27 °C. The bottles were
shook three days apart during the incubation period to
prevent tissue clumping and to promote even and quick
colonisation of the grains by the mycelium (Adeokun et al.,
2012).
B. Preparation of Mushroom Powder
The procedure outlined by Okeke et al. (2003) was
used in the laboratory to create fresh P. ostreatus
mushrooms from fresh mushrooms. They were collected,
cleaned, and then sliced into around 3 mm-thick slices that
were subsequently dried for eight hours at 60°C. The dried
mushroom sample was separately processed in an electric
grinder and sieved through an 80-mesh screen to produce
fine powders. The resulting powder was refrigerated,
hygienically wrapped, and stored in an airtight container for
later use.
Production of Wheat Flour and Mushroom Powder
Biscuit
Mushroom Powder (MP) was substituted for varied
percentages of the wheat flour in biscuits, including 10%,
20%, 30%, 40%, and 50%. The market-purchased 0% MP
biscuit was utilised as the control according to the procedure
of (Bello et al., 2017),
Baking Process
The blended formulations were applied to the baking
process using Chauhan et al. (2012)'s methodology with a
few minor modifications. In the beginning, a Kenwood
mixer (model HM 430) was used to whip the oil and
aspartame till frothy. Skimmed milk and egg white were
added while mixing for around 40 minutes. The right
amount of flour, baking powder, salt, nutmeg, and vanilla
flavour were gradually added to the batter. Then, after
completely combining with water, consistent dough was
made. The acquired dough was kneaded on a smooth, clean
surface for about 5 minutes, thinly flattened with a rolling
pin to a uniform thickness of 5 mm on a wooden board, and
then cut out into the appropriate shapes and sizes. The
dough pieces were cut out, placed on a prepared baking
sheet, and baked for 15 minutes at 160°C. The biscuits were
maintained at 4°C until needed for sensory evaluation and
other studies after being properly cooled and packaged in
airtight polythene. As a control, samples of biscuits made
using white wheat flour were used.
C. Microbiological Analysis
Culture Media and Reagents
The media used for the isolation and enumeration of
the microorganisms associated with the fortified bread are
nutrient agar (NA), which is used for the isolation and
counting of the total viable mesophilic bacterial count, and
potato dextrose agar (PDA), which is used for the isolation
of fungi. According to the manufacturer's instructions, each
medium was sterilised and prepared. The method was used
to manufacture the reagents for the chemical analysis and
biochemical characterization tests in accordance with the
specifications of each analysis Isong et al. (2013).
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Microbiological Examination and Determination of
Total Viable Count
Following baking, the biscuits were stored at 21°C and
50% relative humidity for five days while the physical
changes were noted and the microbiological count was
repeated. This procedure was based on the methodology of
Isong et al (2013). 1g of each sample was taken aseptically
and blended for 2 minutes in 9ml of sterile, distilled water.
Serial dilutions (1ml of each dilution) were used to plate
fungi and bacteria on potato dextrose agar and plate count
agar, respectively, in sterile petri dishes. Bacteria were
incubated for 24 hours at 37 oC and fungi for 8 hours at 20
oC. Using a colony counter (Gallenkamp), visible colonies
were counted and represented as log Cfu/g of the biscuit
sample.
Identification of Bacteria
On fresh nutrient agar, representative bacterial colonies
were chosen and sub-cultured until pure cultures were
obtained. For further research, the colonies were afterwards
kept on nutrient agar slants at a temperature of 5 oC in the
refrigerator. Biochemical tests, morphological traits, and
colony characteristics were used to identify the bacteria.
According to Oyetayo and Oyedeji (2018), morphological
characteristics were observed for each bacterial colony after
24 hours of growth under the microscope while biochemical
characterizations were carried out. The appearance of the
colony of each isolate on the agar media was studied, and
the characteristics observed include: shape, elevation, edge,
optical characteristics, consistency colony surface, and
pigmentation.
Identification of Fungi
On Potato Dextrose Agar (PDA), the microbial
colonies were subcultured. The isolates were recognised
based on their microscopic and morphological
characteristics. A clean glass slide was coated with two
drops of cotton-blue-in-lactophenol, and a little piece of
mycelium free of medium was extracted and applied to the
stain using a sterile inoculating needle. A clean cover slip
was carefully applied over the mycelium after it had been
delicately removed with a needle. This was fixed on the
microscope, and a 100x oil immersion objective lens was
used to study it.
D. Proximate Analysis
Routine analysis of food and food product is termed
the proximate. The carbohydrate determination is given
differences in which the values of crude protein, moisture,
fat, ash and crude fibre is subtracted from 100. The analyses
were carried out on the fortified and unfortified biscuit.
Moisture Content Determination
The Petri dishes were cleaned, properly labelled, dried
in the oven, and weighed (W1). To avoid absorbing
moisture from the air, two gram of each sample were
weighed onto the corresponding plates (W2), spread out
uniformly, and then immediately put into desiccators. The
dishes containing the samples were placed in an oven set to
105 oC for three hours of drying. They were then reweighed
after cooling for 30 minutes in the desiccators. This
procedure was carried out repeatedly until the weight (W3)
became constant. Next, the percentage moisture content was
determined (AOAC, 2012).
Total Ash Determination
Pristine, dried, crucibles were weighed (W1). The
clean, dried, pre-weighed crucibles were filled with about 1g
of each sample, and they were then reweighed (W2). The
crucibles were then heated in the muffle furnace
(Gallenkamp) for three hours at 550 degrees Celsius.
Heating was continued until a light grey or white ash was
formed. The crucibles were taken out of the furnace, brought
to room temperature in desiccators, and weighed (W3).
After reaching a steady weight, cooling and weighing were
both continued (AOAC, 2012). The ash content was
calculated with the formula below;
Crude Fat Determination
1g of each sample was weighed into a filter paper,
which was then weighed (W1) and weighed (W2) after
being neatly wrapped in thread. Petroleum ether (b.pt 40-
60oC) was poured into a round bottom flask until it reached
the third-quarter mark. Reflux condenser was fixed to the
Soxhlet extractor, and the heat source was changed such that
the solvent slowly boils. Petroleum ether (40–60% boiling
range) was used for the 6-hour extraction process under
reflux after the filter paper containing the sample was placed
inside the soxhlet device. After the extraction was complete,
the filter paper and its contents were dried for an hour at
100°C in an oven, then chilled in a desiccator and weighed
again (W3) (AOAC, 2012).
Crude Protein Determination
About 1g of sample was weighed into 50 ml micro
Kjehldal digestion flask and 15 ml of concentrated H2SO4
was added into the flask and a tablet of selenium catalyst
was also added. In the block digester of a fume cupboard,
the mixture was heated to 105 oC until a clear solution was
produced. To ensure that the digest was clear, the flask was
periodically spun. After allowing the digest to cool, the
solution was diluted to 50 ml with distilled water, and 5 ml
of this was added to the distillation apparatus. A 100 ml
conical flask (the receiver flask) was filled with 5 ml of 2%
boric acid before 5 drops of mixed indicator (0.016 g methyl
red + 0.083 g bromocresol green in 100 ml alcohol) were
added. The reaction vessel's digest was added 1.5 ml of 4%
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NaOH through a funnel to make sure it was alkaline and
create a murky solution. All of the exits were closed to
prevent suckback as steam from the steam generator was
delivered into the reaction vessel. With the delivery tube
below the acid level in the receiver flask, the distillation was
conducted into the acid solution. The receiver flask's pink
solution changed to blue during distillation, indicating the
presence of ammonia. Up until 50 cc of distillate were
collected into a receiving flask, the distillation process was
continued. The distillate was then titrated to a pink end point
against 0.1M HCl. The total nitrogen content was calculated
as:
Crude Fibre Determination
A 50ml conical flask that was clean, dry, and well
labelled was filled with one gram of the sample and weighed
(W1). The sample in the conical flask received 200
millilitres of 1.25% H2SO4 and cooked for 30 minutes. The
solutions were thoroughly washed with hot distilled water
after filtering (to remove fat and sugar). With a spatula, the
residue was scraped back into the conical flask. Each sample
then received 200 ml of a 1.25% NaOH solution, which was
brought to boil for 30 minutes. The thoroughly cleaned
residue from each sample was scraped into a crucible, dried
in an oven at 105°C, cooled in a desiccator, and weighed
(W2). The boiled sample was then filtered through muslin
cloth, rinsed twice with industrial methylated spirit and once
more with 10% HCl. The sample was subsequently heated
to 500°C for three hours in the muffled furnace
(Gallenkamp). The samples were taken out of the
desiccators, chilled, and weighed once more (W3). (AOAC,
2012)
Carbohydrate Content Determination
The Nitrogen-free Extractive (N.F.E.), also known as
soluble carbohydrate, is produced using difference rather
than being determined directly.
100 - (% Ash + % Crude Protein + % Crude Fat + %
Crude Fibre + % Moisture) = % Carbohydrate
E. Determination of Mineral Elements
Wet aching was used to determine the mineral content
(potassium, sodium, calcium, magnesium, zinc, iron, and
copper) of each sample, and the mineral content was then
measured using a spectroscope. One gramme of each
sample, in triplicate, was heated to 450 degrees Celsius in a
muffle furnace for 5 to 6 hours. The samples were removed
from the ash and silica dishes and put into the desiccators to
cool before being dissolved in 1ml of 0.5% HNO3. A little
amount of distilled water was combined with number 43
Whatman filter paper before being filtered into a clean,
compact plastic bottle. Atomic adsorption
spectrophotometer (Buck 201, VGP) was used in
determining the mineral content (AOAC, 2012). The
mineral content was calculated using the formula below:
Where R = Solution concentration,
V = Volume of sample digested,
D = Dilution factor,
Wt = Weight of sample
F. Sensory Evaluation of Biscuit
The biscuit products were subjected to organoleptic
analysis using the method described by Oyetayo and
Oyedeji, (2018). A total of 10 untrained panelists consisting
5 males and 5 females, within the age range of 19 – 29 were
drawn from students of Federal University of Technology,
Akure who familiar with biscuit, participated in the
evaluation.
The samples were divided into equal portions and
coded before being presented to the panellists at room
temperature (28±2 oC) under lit fluorescent fixtures. The
panellist consumed the biscuit while rating each sample on a
9-point Hedonic scale (Larmond, 1977), with 1 denoting a
strong dislike and 9 denoting a strong liking. The qualities
that are assessed include acceptability overall, colour,
texture, taste, and odour.
G. Statistical Analysis
All experiments were carried out in triplicate. One-way
analysis of variance (ANOVA) was used to analyse the data,
and Duncan's New Multiple Range test (SPSS 16.0 version)
was used to compare the means. At a p-value of 0.05,
differences were deemed significant.
III. RESULTS
A. Microbiological Examination and Determination of
Total Viable Count
The result of the microbiological examination of total
viable bacteria count of mushroom biscuit composition is
shown in Figure 1. The composition with 90% mushroom
had the highest bacteria count followed by the composition
with 60% mushroom (60/40). The least viable count was
recorded in the 50/50 and 100% composition respectively.
B. Biochemical Identification of Bacteria
The result for the biochemical characteristics of
bacterial isolates is shown in Table 1. The organisms
tentatively identified after biochemical test includes;
Staphylococcus warnei, Bacillus niacin, Kluyuera
georgiana, Bacillus circulans, Bacillus licheniformis,
Bacillus megaterium, Bacillus barbaricue, Bacillus cereus,
and Bacillus simplex. The most prominent of the organisms
isolated was Bacillus sp.
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C. Macroscopic Identification of Fungi
Table 2 shows the macroscopic identification of fungi and the fungal genera isolated were identified as Aspergillus flavus, A.
niger, and Pleurotus ostreatus.
Fig 1 Microbiological Examination of Total Viable Bacteria Count of Various Mushroom Biscuit Compositions
Table 1 Biochemical Identification of Bacteria
Keys: + = Positive; - = Negative
Table 2 Macroscopic and Microscopic Features of Fungal Isolates
S/N
Macroscopic Description
Microscopic characteristics
Fungi
1
Black and powdery colonies
Conidiophores terminates in vesicles, smooth walled colorless with
brownish shade
Aspergillus niger
2
Yellowish green colonies
Conidiophores are coarsely roughened, uncolored with vesicles,
spherical and metulae covering nearly the entire vesicles
A. flavus
3
Thick, Smooth, Cream,
broad fan shaped
In clusters and interwoven with whitish gills running down and a
nearly absent stem
Pleurotus
ostreatus
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D. Proximate Analysis of Mushroom
The result for the proximate composition for the total
mushroom below in Figure 2 showed that carbohydrate is
the highest composition at 73%, while ash content was the
lowest at about 2%. Fibre content also contributes
significantly to the mass of the mushroom at 17%, while
moisture and nitrogen were measured at 10% and 8%
respectively.
Moisture Content Determination
The result of the determination of the moisture content
of the different mushroom biscuit sample formulations in
Figure 3 showed that sample C (70% mushroom) had the
highest moisture content, while sample A (80% mushroom)
had the significantly lowest even when compared with the
control sample F (100% mushroom). Meanwhile Sample B
(60% mushroom) and D (50% mushroom) had close range
of moisture content.
Total Ash Determination
The result of the determination of the ash content of
the different mushroom biscuit sample formulations showed
that the addition of mushroom to the biscuit composition
generally increased the ash content of the resultant biscuit
with sample D (50%) mushroom having the highest ash
content, while control group i.e., sample F (100%
mushroom) had the least observed ash content (Figure 4).
Crude Fat Determination
Figure 5 presents the result of the determination of
crude fat content for different mushroom biscuit sample
formulations. It showed that there was only little difference
between the fat contained within all the various mushroom
biscuit formulation. However, sample D (80%) and B (60%)
mushroom showed the same and the lowest content of fat
when compared with the control group (0%) mushroom.
Crude Protein Determination
The result of the determination of the protein content
of the different formulations of biscuits fortified with
mushroom as presented in Figure 6 showed that sample B
(60% mushroom composition) had the highest protein
content, which was followed by sample C (70%), while the
least was observed in sample E (90% mushroom
composition). Generally, the result showed that the higher
the mushroom content, then the lower the protein content
estimated in the mushroom biscuit.
Crude Fiber Determination
The result of the determination of the fiber content of
the different formulations of biscuits fortified with
mushroom when compared with the control group which
had (0%) mushroom composition showed that the addition
of mushroom to the biscuit greatly increased its overall fibre
content. However, the highest crude fibre content was
observed in sample D which had equal mushroom and
flower content at 50% each (Figure 7).
Carbohydrate Content Determination
The result of the determination of the carbohydrate
content of the different formulations of biscuits fortified
with mushroom showed that carbohydrate content of the
mushroom decreased with increased addition of mushroom
to its formulation with the slight exception of sample D
(80% mushroom composition), while the carbohydrate
content level of other percentages in A, B, C, and E were
observed to be extremely low (Figure 8).
Fig 2 Percentage Proximate Composition of Mushroom
Pleurotus oestratus
Fig 3 Percentage Content of Various Mushroom Biscuit
Formulations
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Fig 4 Percentage Ash Content of Various Mushroom Biscuit
Formulations
Fig 5 Percentage Fat Content of Various Mushroom Biscuit
Formulations
Fig 6 Percentage Protein Content of Various Mushroom
Biscuit Formulations
Fig 7 Percentage Crude Fiber Content of Various Mushroom
Biscuit Formulations
Fig 8 Percentage Carbohydrate Content of Various
Mushroom Biscuit Formulations
Determination of Mineral Elements
Figure 9 present the result of the determination of the
mineral composition of the different formulations of biscuits
fortified with mushroom, it showed that the most abundant
mineral in the biscuit are Na, K and Ca while Fe and Pb
were low significantly. Potassium (K) was the overall
highest mineral content in the fortified biscuits, with sodium
(Na) observed to also have the second overall highest for
mineral nutrient content and generally, lead (Pb) was only
present in trace amount. 50% mushroom biscuit formulation
was generally observed to contain the highest potassium and
sodium content at 19.5±0.5 and 10.15±0.15 ppm
respectively. The calcium content of fortified mushroom
biscuit showed that no significant increase when compared
with the control (0% mushroom).
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Sensory Evaluation of Biscuit
The result of the determination of the sensory evaluation of the various formulations of biscuits fortified with mushroom in
Table 3 showed that sample E (10% mushroom) had the best overall acceptability at 9.50±0.17, with texture and taste both at
9.30±0.21 which was only a little lower than the values for standard biscuits (control) on a general measurement scale of 1-10.
Sample A containing 50% mushroom had the poorest acceptance at 5.40±0.85. However, sample D (20% mushroom) had the best
aroma production valued at 9.10±0.28.
Fig 9 Mineral Composition of Various Mushroom Biscuit Formulations in Ppm
Table 3 Sensory Evaluation of Mushroom Biscuit
Quality evaluated
Samples
Aroma
Colour
Texture
Taste
Overall acceptability
A
8.10±0.43
7.40±0.45
7.10±0.59
5.50±0.40
5.40±0.85
B
8.30±0.30
7.80±0.29
7.90±0.57
6.30±0.52
5.90±0.86
C
7.90±0.91
8.90±0.28
8.40±0.22
6.90±0.87
7.40±0.92
D
8.50±0.31
9.10±0.28
9.40±0.22
7.80±0.39
8.80±0.20
E
8.80±0.33
8.40±0.31
9.30±0.21
9.30±0.21
9.40±0.16
F
8.00±0.42
9.20±0.29
9.60±0.16
9.50±0.22
9.50±0.17
Key: Mushroom compositions, A; (50/50), B; (60/40), C; (70/30), D; (80/20), E; (90/10) and F; Control (100/0)
IV. DISCUSSION
Baked products such as biscuits is among the highly
consumed products acceptable by all (Wan Rosli et al.,
2012) due to the fact that they are popular, convenient,
inexpensive and its long shelf-life, however the primary
ingredient (wheat) used in its production is deficient in
protein and some other essential nutrients. A study by
Nieburg (2012) have shown that replacing the wheat flour
with mushrooms at different levels in formulations could
have a substantial improvement in the protein content and
other essential nutrients such as the vitamins, minerals and
dietary fiber without affecting its physical and sensory
properties.
From this study, 4 bacteria genera (Bacillus spp,
Pseudomonas spp, Staphylococcus spp and Kluyvera) and
fungi (Aspergillus flavus and Aspergillus niger) were
isolated from the cultivated mushroom (Pleurotus
ostreatus), however the dominant bacteria is the bacillus
spp. This observation is similar to Mariusz and Stanislaw
(2019) who also recorded a high number of Bacillus spp.
This could be attributed to fact that Bacillus and
Pseudomonas are growth promoting bacteria due to their
competitive and antagonistic activity against several
Volume 8, Issue 10, October – 2023 International Journal of Innovative Science and Research Technology
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IJISRT23OCT1320 www.ijisrt.com 1384
pathogens (Company et al., 2010). Some of the isolates
could also be associated with the substrate on which the
mushroom was cultivated as it is related to the report of
Obire and Amadi (2013) who (similarly isolated Aspergillus
and Mucor spp. from fermented sawdust) isolated from the
substrate in which the mushroom was grown.
The proximate composition of the Pleurotus ostreatus
shows that it has percentage protein and crude fiber of 21%
and 10% respectively. Singh and Thakur (2016) also
reported similar work, having a 13% protein content,
however Oyetayo and Ariyo (2013) and Bello et al. (2017)
reported a higher percentage of protein (20- 24%) and crude
fiber (17-30%) respectively in P. ostreatus cultivated on
different wood substrates. The variation in nutrient from
different study could be due to the nutrient composition of
the substrates on which the mushrooms was cultivated. The
protein, crude fiber and ash content from the proximate
analysis of the biscuit from this study is similar to the low
values of (10, 2 and 1% respectively) reported by
Adegbanke et al. (2020) in wheat biscuit.
The proximate composition of biscuit fortified with
Pleurotus ostreatus showed increase in moisture content,
total ash, crude protein and a significant increase in the
crude fiber, no significant increase was seen in the fat
content compare with the control; however, a decrease was
observed in the percentage carbohydrate. Bello et al., (2017)
and Prodhan et al., (2015) also recorded a similar trend of
proximate composition of Pleurotus sajur-caju fortified
biscuit.
The protein content at different percentage of
mushroom flour inclusion ranges from 9% to 15% and a
control of 8.4%. There was a significant increase in all the
samples compared with the control. The increase in the
protein content could be attributed to the high protein
content of the mushroom added and this reveals that the
fortified biscuit would be a better-quality compare with the
unfortified one (Bello et al., 2017). Also, in agreement is
Ayo et al. (2014) who enriched a malted soy biscuit,
however Mepba and Iboh (2007) reported a decrease in
protein content in wheat plantain biscuit. The significant
increase in the crude fiber compares with the control ranges
between 6.5% to 10% which could be attributed to the
mushroom used, been a rich source of fiber which
eventually increases the fiber content of the biscuit with
increase in the level of addition. Bello et al. (2017) reported
that a high intake of dietary fiber helps prevent constipation,
reduce cholesterol level in the blood, slow digestion and
sudden release of energy thus making blood level stable.
The ash content of a food material could be used as an index
of mineral constituents of the food; therefore the increase in
the ash content at all level of addition indicates the enriched
biscuit to be a good source of mineral.
The carbohydrate content was found to decrease with
increase in the mushroom flour, however there was no
significant decrease in the (80/20) concentration compare
with the control. A similar trend was presented by Ng et al.
(2017) and Nordiana et al. (2019) who fortified biscuit with
mushroom flour, however Aishah (2013) documented that
the carbohydrate content increases with increase in
mushroom flour, though this may be due to the different
formulations of the cakes. The moisture content of biscuit
and mushroom pre-determined before mixing was 10.4%,
the mixture after production has increased moisture content.
Aishah (2013) and Nordiana et al. (2019) has documented
similar concurrent increased moisture content, this could be
that the mushroom flour possess high water content and
water holding capacity, also the sugar, starch and dietary
fibre in the formulation could have also contributed as they
contain high amount of water (Mohammed et al., 2010) In
contrast to this report is Ng et al. (2017) who recorded a
lower moisture as compared to the control , this may be as a
result of the very high temperature used in baking thereby
reducing the water content.
The most abundant mineral in the biscuit are Na, K and
Ca while Fe and Pb were present in trace amount.
Adegbanke et al. (2020) also reported Na, K, Ca, P and Zn
in significant amount and Cu, Pb, Fe and Mn were reported
to be found in trace amount in wheat biscuit. The mineral
composition of the fortified biscuit shows significant
increase in Na and K, a slight increase was also observed in
Fe content while a general decrease was observed in the Ca
content and Pb remain insignificant. The mineral
composition obtained from this study shows that the
fortified biscuit is a good source of mineral. The most
abundant mineral is K followed by Na, this is in line with
the report that the most abundant mineral in biscuit is K and
Na and both are required to maintain osmotic balance in the
body fluid, pH of the body, to regulate muscle and nerve
irritability, control glucose absorption and enhance normal
retention of protein during growth (Arinathan et al., 2003).
The fortified biscuit is also a good source of Fe, which is a
major component of hemoglobin that carries oxygen to all
part of the body and it also plays a critical role in overall cell
function (Bello et al., 2017). The decrease in the Ca content
when compared with the control, indicates that the fortified
biscuit is not a good source of Ca and Ca is involved in the
overall body formation and functions. The results of Bello et
al. (2017) appears similar with the result of this study’s
mineral content, however it recorded a steady decrease in
Na and then a slight increase in the 30% addition of the
mushroom flour.
The result of the sensory evaluation of the fortified
biscuit after production indicates that the biscuit fortified
with 10% and 20% mushroom flour does not show any
significant difference from the control, while there is a
significant difference between the control and the biscuits
fortified with 30%, 40% and 50% mushroom flour and the
overall acceptability of the fortified biscuit decreases with
increase in the mushroom flour. This could be as a result of
the obvious and unpalatable taste from adding larger portion
of the mushroom flour, this aligned with the study of
Prabhasankar et al. (2009), Aishah (2013) and Bello et al.
(2017) who reported that the panelist preferred lower
percentages of oyster mushroom flour, and sensory scores
were reduced significantly when more mushroom powder is
added as compared to the control.
Volume 8, Issue 10, October – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
IJISRT23OCT1320 www.ijisrt.com 1385
V. CONCLUSION
The study suggests that the nutritional value of wheat
biscuits enriched with mushroom (P. ostreatus) improved
with the addition of more mushroom powder. However,
with high addition levels of the mushroom powder, the
general acceptance of the enriched biscuit declines. For the
optimum nutritional, essential mineral, and sensory
qualities, it is advised that commercial biscuit manufacture
be done at a 50% level of mushroom inclusion. The
production of biscuit using other types of flour in synergy
with mushroom (P. ostreatus) could be studied to determine
its nutritional value and sensory acceptability warrants
industrial application. Malnourished persons, communities
and societies can be provided these fortified biscuits to
alleviate consumption of foods with low levels of essential
nutrients because of its high nutritional composition.
Authors’ Contribution
Author Victor O. Oyetayo supervised the study and
provided administrative support. Author Rosemary A. Ajayi
performed the literature search, laboratory work, data
analysis and wrote the first draft of the manuscript. All
authors read and approved the final draft.
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