Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
ISSN: 2073-9524
eISSN: 2310-8746
34
Utilization of banana puree fermented by Lactobacillus
acidophilus LA5 and Bifidobacterium lactis BB12 for the
manufacture of synbiotic ice cream
Abdullah Ismaeel Awad
1
, Amer Hussein Hamdan Alzobaay
2
1,2
Department of Food Science, College of Agricultural Engineering Sciences, University of
Baghdad, Iraq.
1
Corresponding author: abdullah.sumerian@gmail.com
Article history:
Received: 2 December 2022
Accepted: 19 January 2022
Published: 30 June 2023
Abstract
In this study, manufacturing synbiotic ice cream is containing fermented banana
puree by Lactobacillus acidophilus LA5 and Bifidobacterium (Bif) lactis BB12
and probiotic ice cream containing fermented skim milk by the same bacteria
under the same conditions. The Viability of probiotic starters, pH and sensorial
properties were tested during the storage period, in addition to the overrun.
Synbiotic ice cream showed high bacterial count for both species of probiotics,
especially Ice.4 treatment (ice cream containing 10% fermented banana puree
with same couple strains) which was 7.54 and 7.59 logarithmic (log) colony-
forming unit (CFU)/ gram (g) for Lactobacillus (Lb). Acidophilus LA5 and Bif.
lactis BB12, respectively, pH registered 5.7, sensorial attributes recorded 84%
and overrun reached 78% compared to others trials that inoculated with 5% of the
same puree or ice cream with fermented skim milk with (5,10) % or control
treatment (without starters).
Keywords: probiotics,
prebiotics, lactic acid
bacteria, sensory
evaluation, overrun.
https://dx.doi.org/10.52951/dasj.23150105
This article is open-access under the CC BY 4.0 license (http://creativecommons.org/licenses/by/4.0/).
Introduction
Ice cream is a widely consumed product
around the world. It is frozen foam of a
mixture that includes water, air, non-fat
solids, emulsifiers, fat, flavors, colorants
and stabilizers (El–Samahy et al., 2009).
Probiotics defined as are ―live
microorganisms that, when administered in
adequate amounts, confer a health benefit on
the host‖, as the addition of probiotics to
food increases the nutritional and functional
value of products by enhancing the quantity
and availability of nutrients and bioactive
compounds arising from microbial
metabolism, including organic acids,
conjugated linoleic acid and
exopolysaccharides. These microorganisms
can be enhanced via a substrate that
consumes it selectively can confer a health
benefit called prebiotics (Villalva et al.,
2017). There are many probiotics strains
belonging to Lactobacillus,
Bifidobacterium, some of the Bacillus
species, the yeast Saccharomyces cerevisiae,
and others that are known generally
recognized as safe (GRAS) (Rasheed et al.,
2020). The strains belonging to the genus
Lactobacillus and Bifidobacterium are
among the most prominent species of
probiotics added to ice cream and other
dairy products. The combination of
probiotics and prebiotics is called
"synbiotics", it causes a synergistic effect
that stimulates the growth of probiotics
through the fermentation of prebiotics
(Villalva et al., 2017). The combination of
probiotics and prebiotics improves the
survival of the probiotics in the digestive
tract as it enhances tolerate low pH, oxygen
and temperature conditions (Al-Hassani and
Mustafa, 2022).
Enriching the dairy products such as
yogurt and milk with probiotics may achieve
beneficial health effects, most notably
reducing the level of cholesterol in the
blood, contributing to the prevention of
mailto:abdullah.sumerian@gmail.com
http://creativecommons.org/licenses/by/4.0/
https://orcid.org/0000-0003-0969-5745
https://orcid.org/0000-0003-4919-4099
https://orcid.org/0000-0003-0969-5745
https://orcid.org/0000-0003-4919-4099
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
35
colon cancer, relieving the pain of irritable
bowel syndrome (IBS) and increasing
antioxidants and inhibition compounds
(Niamah et al., 2017). Probiotics can also
contribute to the prevention of ulcers,
gastrointestinal infections, diabetes, heart
disorders, and vitamin B production and
stimulate the immune response (Mohammed
et al., 2021). Fermented foods with
probiotics, such as yogurt, are also a suitable
option for people who suffer from lactose
intolerance, due to the ability of probiotics
to produce high levels of the enzyme
lactase, it also can modify the metabolic
activities of colonic microbiota and thus
alleviate lactose intolerance (Ibrahim et al.,
2021). When the arrival of probiotics to the
digestive system of the host in sufficient
quantities, they adhere to the special
receptors on the surface of the intestinal
epithelial cells, which prevents the adhesion
of harmful microorganisms, which leads to
excretion it outside the body. Thus,
strengthening the beneficial bacteria that are
mainly present in the gastrointestinal tract,
and enhancing the body’s immunity to
confront pathological infections (Jasim and
Taha, 2017). Recently, the enrichment of
certain bacterial population in the digestive
system with probiotics, prebiotics, and
symbiotics has become alternatives to
antibiotics (Aziz and AL-Hawezy, 2022).
The nutritional composition of banana
puree presents a suitable substrate for the
fermentation of probiotic products (Gallo et
al., 2021). Bananas are a common source of
prebiotics that promote the growth of
probiotics, as contain a sufficient amount of
resistant starch, oligofructose, inulin, and
others (Sidhu and Zafar, 2018). Therefore,
the study aimed to produce a synbiotic ice
cream that includes fermented banana puree
via the probiotic Lactobacillus acidophilus
LA5 and Bifidobacterium lactis BB12 and
compare it with the probiotic ice cream
includes the same probiotic species through
different treatments with determining of the
trail that ensures high viability of probiotics,
appropriate pH, good overrun and better
sensory acceptance of the product to provide
a desirable healthy product and at the same
time achieve an adequate economic profit.
Materials and Methods
Commercial lyophilized cultures of the
probiotics Lactobacillus acidophilus LA-5
and Bifidobacterium lactis BB12 (supplied
by Chr-Hansen) were activated with MRS
broth medium three times and then activated
with skim milk until the initial counts
reached 11.45 and 11.42 log CFU/ml for
Lactobacillus acidophilus LA-5 and
Bifidobacterium lactis BB12, respectively.
Green bananas were purchased from a
local market in Baghdad, Iraq, the puree was
produced from the pulp of the fruit. Banana
fruits were wiped with ethyl alcohol (70%),
then they were peeled, and cut into small
pieces (transverse slices), the slices were
dipped in 0.5% (by mass per volume) citric
acid solution for 10 min. then mixed using
an electric mixer in order to obtain a
homogeneous mixture, and then the puree
was distributed in sterilized sealed glass
cups (Alzobaay et al. 2013). Inoculation of
banana puree by probiotic mixed cultures of
Lactobacillus acidophilus LA-5 and
Bifidobacterium lactis BB12 (50:50) using
10 ml and incubation at temperature 37 °C
for 48 hours until the initial counts reached
11.37 log CFU/g.
Ice cream mixture treatments were
prepared by Mahmedand and Obeed’s
(2020) method. The ingredients used in
making 1 kg of ice cream mixture included:
705 g full cream milk powder (NIDO/the
French company Nestle), 130 g fresh cream
(40% fat) (Dairy plant of College of the
Agricultural Engineering Sciences/
University of Baghdad), 160 g table sugar
(sucrose) was used as sweetener, and 5 g
CMC (Carboxymethyl cellulose) stabilizer
was used in the production of ice cream
(10% fat). The ice cream mixtures were
modified to include 5 and 10% fermented
skim milk for the probiotic ice cream
treatments (Ice.1 and Ice.2), respectively, as
well as 5 and 10% fermented banana puree
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
36
for the synbiotic ice cream treatments (Ice.3
and Ice.4), respectively. Ice cream mixtures
were incubated at 37 °C until the pH
reached 5.90. Then cooled to 5°C, and left at
5°C overnight for aging. The mixtures were
then frozen for 20 min in a CARPIGIANI
freezer (Italy). The ice cream samples were
packed into plastic containers (100 ml), at a
temperature of 5 °C and stored at -18 °C
until analysis (Al-Shawi and Ali, 2020).
Probiotic bacteria enumeration to ice
cream treatments was done using tryptose
proteose peptone yeast extract (TPPY) agar
with added prussian blue which was
prepared from the following ingredients:
glucose (10 g), lactose (10 g), tryptone (7 g),
peptone (7 g), yeast extract (2 g), tween 80
(1 ml), prussian blue (0.3 g), and 15 g of
agar was added to it. All the ingredients
were dissolved in a liter of distilled water. It
was sterilized with an autoclave at 121
ºC/15 minutes, cooled to 50 ºC and then
prussian blue was added. Ten grams of ice
cream were diluted in 100 ml of sterilized
peptone water solution. One ml of the
diluted samples of ice cream treatments
were poured on sterile TPPY-prussian blue
agar. It was used for the differential
enumeration of probiotic cultures.
Incubation of plates was conducted under
anaerobic conditions using a Gas-pack
system at 37°C for 48-72 h. Results of
enumeration were expressed as (log10
CFU/g). Analysis of samples was conducted
in duplicate. TPPY-prussian blue agar was
proposed as a differential agar that allows
Lb. acidophilus and Bifidobacterium to be
enumerated on one medium.
Bifidobacterium gave white colonies and Lb.
acidophilus produced large pale blue
colonies surrounded by a wide royal blue
zone (Ashraf and Shah 2011; Teixeira,
2014).
pH values of ice cream treatments were
measured by pH-meter (Hach Sension+
pH3) with a pH electrode Sension+ 5011T,
firstly pH meter was calibrated using a
buffer of pH 4 and pH 7, then 20 ml of the
melted samples (20 ± 1.0°C) was putted in a
50 ml beaker, as pH values was determined
by the glass electrode. Each measurement
occurred in triplicate (Zagorska et al.,
2022).
The sensory assessments were
conducted in the central laboratory of the
Department of Food Science in the College
of the Agricultural Engineering Sciences/
University of Baghdad. The panelists of
nine raters were experts and well-known in
this field. Panelists were provided with a
glass of water and, instructed to rinse and
swallow water between samples. They were
given written instructions and asked to
evaluate products for acceptability based on
their taste, smell, color, texture and
mouthfeel. Overall acceptability was
determined by a special scale that we
created according to the requirements of the
experiment. Sensory evaluation was
performed using a form modified (table 3)
by Prashanth et al. (2018).
Overrun is the quantity of air absorbed
by the ice cream mixture during
manufacturing. High overrun values appear
in the absorption of a high amount of air
into the ice cream product, thus expanding
the product (Asres et al., 2022). The overrun
values of ice cream treatments were
determined in duplicate based on differences
in weight (Wt.) after and before freezing as
Elkot et al. (2022) stated by using the
following equation:
% Overrun = (Wt. of the mix – Wt. of the
same vol. of ice cream)/Wt. of the same vol.
of ice cream x 100%.
To detect the effect of different factors
in study parameters, The Statistical Analysis
System- SAS (2012) program was used
(Cary, 2012). The least significantly
difference (LSD) test was used to significant
compare between means in this study
Results and Discussion
Probiotics count in ice cream
The results in Table 1 showed no
statistically significant differences (P≤0.05)
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
37
in the viable counts of probiotic ice cream
trials, which included two treatments for
probiotic ice cream (Ice.1) and (Ice.2) with a
concentration of 5 and 10%, respectively,
and two treatments for synbiotic ice cream
(Ice.3) and (Ice.4) at a concentration of 5
and 10%, respectively, during 120 days of
storage.
Synbiotic ice cream treatments
maintained higher viable probiotic counts
than the probiotic ice cream treatments.
Despite the gradual decrease in the viable
counts in all the ice cream treatments, the
decrease was less in the synbiotic ice cream
treatments as compared to the probiotic ice
cream treatments. The reason for this can be
attributed to the fact that synbiotic ice cream
treatments (Ice.3) and (Ice.4) contain 5 and
10% of fermented bananas, respectively, as
banana puree is rich in prebiotics including,
insoluble and soluble dietary fibers (DF),
pectin, inulin and Fructo-oligosaccharides
(FOS) (Kumar et al., 2014) As these
prebiotic compounds support the survival of
probiotic cells. Al-Shawi and Ali, (2020)
indicated the action of inulin as a protective
mechanism for Lb. acidophilus cells during
ice cream storage and FOS promotes the
growth of Lb. acidophilus. Also, Prashanth
et al. (2018) indicated that green bananas
are rich in resistant starch, as resistant starch
can act as a protective factor for probiotic
cells during ice cream frozen storage.
The same applies to the viable counts of
Bifidobacterium lactis BB12 bacteria, which
were found to maintain their viability more
in synbiotic ice cream treatments compared
to probiotic ice cream treatments due to the
effect of prebiotics sourced from fermented
banana puree. The results of our study are
consistent with Shori (2021), who
mentioned that the addition of 2% inulin to
ice cream containing 10% w/w fermented
milk enhances the viability of Bif. lactis. It
was observed that the sample (Ice.4) which
included 10% fermented banana, was the
best treatment for obtaining synbiotic ice
cream with high viable probiotic counts,
most likely due to the increase in the
proportion of fermented banana puree added
to the ice cream compared to the sample
(Ice. 3) which included 5% fermented
banana. This means better support for the
probiotic cells in the ice cream matrix due to
the high proportion of probiotics. This
assumption is supported by Krawęcka et al.
(2021) who reported that adding bananas to
ice cream helps maintain the viability of
probiotics during freezing due to their
polysaccharide content.
In most treatments, it was noticed that
the counts of Bif. lactis BB12 were slightly
lower than those of Lb. acidophilus, because
Bifidobacterium spp. are strictly anaerobic
cells, and ice cream is an environment that
contains a rich concentration of oxygen,
which makes the cells of these bacteria
vulnerable to dissolved oxygen, which leads
to the accumulation of toxic metabolites,
including superoxide, hydroxyl radicals and
hydrogen peroxide, hence causing cells
death. Bifidobacterium spp is more
susceptible to these toxic factors than Lb.
acidophilus (Boza-Méndez et al., 2012).
However, it was observed that there was a
gradual decrease in the count of viable
probiotics with continued storage for 120
days because of the effect of freezing injury
that leads to cells' death (Al-Shawi and Ali,
2020). Also, the effect of low temperatures
and high concentrations of dissolved oxygen
makes it difficult for probiotics to increase
their count in ice cream during freezing
storage (Boza-Méndez et al., 2012).
Andriani and Wikandari (2022) support
our finding as they stated the viability of
lactic acid bacteria (LAB) can be maintained
by adding skim milk to the product, for it
can act as cryoprotectant (protective
bacteria) during freezing. Our study showed
that the counts of probiotics for both species
were higher in the sample (Ice.2), which
included 10% of the probiotic skim milk,
compared to the sample (Ice.1), which
included 5% of the probiotic skim milk. The
increase in the proportion of the probiotic
skim milk added to the ice cream mixture
provides better protection for the probiotic
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
38
cells during the frozen storage of the
product. Abdulrazzaq and Khalil (2022)
mentioned that skim milk can act as a
prebiotic source for probiotic bacteria. Lb.
acidophilus is generally more tolerant to
acidic conditions than Bifidobacteria whose
growth retards at a pH less than 5.5. The
tolerance of Bifidobacteria to acidic
conditions varies according to the specific
strain (Mohammadi et al., 2011). It was
found that Lb. acidophilus has a such high
acid activity that it can tolerate higher acidic
conditions than Bif. lactis (Zomorodi, 2019).
Results agree with Nunes et al. (2018) who
reported that Lb. acidophilus La-5 (ML)
presented higher survival than Bif. Bb-12
(MB) under these conditions.
The results showed there was a decrease
in the counts of probiotics after frozen
storage but still in a fairly high amount. It
met the standard as a probiotic product
according to Andriani and Wikandari,
(2022) stating that the minimum amount of
LAB contained is 10
7
CFU/g.
It is noted that the survival of probiotics
during the storage period which lasted 120
days was without growth as viable cells in
sufficient numbers with the possibility of
cultivation when the appropriate conditions
are available, as a result of the ability of
probiotic species to adapt in these conditions
by regulating the osmotic stress in the
presence of sugars like sucrose and lactose.
In addition to the role of prebiotics in
forming a matrix that protects probiotic
cells, it reduces the damage caused by
freezing (Tripathi and Giri, 2014).
Freezing causes a reduction in the cells’
vital metabolic activities, and therefore cell
growth does not occur under these
conditions. Freezing leads to the death of
some of the cells, but other numbers of cells
resist freezing. The nature of damage to
freezing depends on the conditions and
velocity of the freezing process and the
presence of protective materials from
freezing such as prebiotics. The resistance
of probiotics to the damage of freezing also
varies between strains of probiotics. Also,
one of the studies mentioned that the death
rate of probiotics cells was greater during
the freezing process than during storage.
Probiotics show better survival in freezing
conditions if they can dehydrate without
breaking their cell membranes. Previous
studies showed that the two strains of
probiotics Lactobacillus acidophilus La-5
and Bifidobaterium lactis Bb-12 used in our
study achieve high survival rates in ice
cream (Mohammadi et al., 2011)
Table 1. Viable probiotics of ice cream treatments during the freezing storage period
Logarithm of Probiotics bacteria counts (CFU/g)
Days
LSD
value
Lactobacillus acidophilus LA-5 LSD
value
Bifidobacterium lactis BB12
Ice.4 Ice.3 Ice.2 Ice.1 Ice.4 Ice.3 Ice.2 Ice.1
0.48 NS 8.07 7.88 8.06 7.82 0.43 NS 8.05 7.83 8.01 7.7 1
0.41 NS 8.02 7.85 8.04 7.79 0.48 NS 8.03 7.75 7.98 7.70 15
0.37 NS 7.98 7.81 7.95 7.70 0.39 NS 7.96 7.69 7.93 7.66 30
0.38 NS 7.94 7.71 7.91 7.65 0.42 NS 7.89 7.62 7.87 7.59 45
0.38 NS 7.78 7.59 7.80 7.49 0,38 NS 7.76 7.56 7.62 7.53 60
0.41 NS 7.72 7.53 7.61 7.44 0.44 NS 7.65 7.54 7.55 7.49 90
0.39 NS 7.54 7.46 7.43 7.41 0.40 NS 7.59 7.43 7.47 7.32 120
NS: Non-Significant
NS: Non-Significant statistical differences, Ice.1 and Ice.2 are probiotic treatments including probiotic skim
milk in the ratio 5% and 10%, respectively. Ice.3 and Ice.4 are synbiotic treatments including banana pulp puree
in the ratio 5% and 10%, respectively.
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
39
pH values of ice cream
Results presented in Table 2 showed the
difference in the pH values of the probiotic
and synbiotic ice cream treatments on the
first day, it is due to the addition of probiotic
skim milk to the ice cream by 5% to
treatment (Ice.1) and by 10% to treatment
(Ice.2). The pH reached 5.36 in treatment
(Ice.1) and decreased to 5.29 in treatment
(Ice.2), and this finding is supported by
Andriani and Wikandari (2022), which can
be explained due to the accumulation of
fermentation products, especially organic
acids such as lactic and acetic acid.
The pH in (Ice.3) and (Ice.4) decreased
to 5.89 and to 5.72 successively because of
adding 5% and 10% of fermented banana
puree to each treatment. This finding is
supported by Prashanth et al. (2018). They
emphasized pH increase as more green
banana flour was added to the ice cream
because it contains organic acids. A slight
low decrease in the pH values was observed
after 45 days of frozen storage in the
probiotic and synbiotic ice cream treatments
that started at different pH values for each
treatment and this could be attributed to the
decrease in the viable counts of probiotics
due to the freezing effect due to the
cessation of metabolic activities.
The results of our study also agree with
the study of Hasan et al. (2020) who
indicated that replacing part of the skim
milk with different types of dried fruit
powder (including dried banana powder)
resulted in a lower pH value for all
treatments compared to the control group.
Gheisari et al. (2016) found a gradual
decrease in the pH of the probiotic ice cream
containing Lb. casei during frozen storage
for three months from 6.56 on the first day
to 6.47 on the 90
th
day attributing the reason
to the metabolic activity of the microflora
by Embden- Meyerhof-Parnas pathway
leading to the production of organic acids
such as lactic and/or acetic acid. A decrease
in pH in ice cream containing probiotics
during storage, and the addition of dietary
fiber and culture increased the acidity and
decreased pH (Afzaal et al., 2020).
Table 2. pH values of ice cream treatments during storage
pH values during storage (days) Samples
120 90 60 45 30 15 1
6.82 6.82 6.82 6.82 6.82 6.82 6.82 Control Ice.
5.33 5.33 5.34 5.34 5.35 5.35 5.36 Ice.1
5.27 5.28 5.28 5.28 5.29 5.29 5.29 Ice.2
5.88 5.88 5.88 5.88 5.89 5.89 5.89 Ice.3
5.70 5.70 5.70 5.71 5.71 5.71 5.72 Ice.4
1.26 * 1.22 * 1.22 * 1.14 * 1.06 * 1.18 * 1.04 * LSD value
* (P≤0.05)
* Statistically significant differences (P≤0.05), Ice.1 and Ice.2 are probiotic treatments including probiotic skim
milk inthe ratio 5% and 10%, respectively. Ice.3 and Ice.4 are synbiotic treatments including banana pulp puree
in the ratio 5% and 10%, respectively.
Sensory evaluation of ice cream
Table 3 showed statistically significant
differences (p≤0.05) in the results of sensory
acceptance for all ice cream treatments
during the freezing storage period for the
first day, day 60 and day 120, respectively.
Regarding the taste and smell of the
synbiotic ice cream, it is noted that the
highest sensory evaluation scores were in
the trail (Ice.4), reaching 18 for both
features from the first day to the end day of
frozen storage. It is higher compared with
the trail (Ice.3). Hasan et al. (2020)
indicated that replacing part of the skim
milk with different types of dried fruit
powder (including dried banana powder)
scored high points for flavor, and the flavor
score increased with the increase in the
replacement rate due to the addition of more
of fruit flavor.
Banana can improve the flavor of ice
cream as containing inulin. Ozturkoglu-
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
40
Budak et al. (2019) stated that the addition
of inulin in fermented milk supports an
increase in the concentration of lactic and
acetic acid from the probiotics Lb.
acidophilus La-5 and Bif. animalis Bb-12
because inulin has a stimulating effect on
the growth and viability of probiotics.
As for the probiotic ice cream, it is noted
that the sensory evaluation scores were
highest in the trail (Ice.2), reaching 18 from
the first day to the end day during frozen
storage. It is higher compared with the trail
(Ice.1), and this difference is attributed to
the high proportion of probiotic skim milk
in the trail (Ice.2) which amounted to 10%
compared to the trail (Ice.1) which included
5% of probiotic skim milk.
Probiotic and synbiotic ice cream trails
showed an improved taste compared with
the control treatment for several reasons,
most notably the presence of Lb.
acidophilus, which gives an umami taste
and contributes significantly to the acid taste
due to lactic acid production (Cui et al.,
2019). Studies showed that the volatile
compounds concentrations of acetaldehyde,
acetone, and diacetyl were lower in the
concurrent products supplemented with
inulin, compared to the non-supplemented
probiotic products (Ozturkoglu-Budak et al.,
2019).
Ice.4 sample registered high overall
acceptance scores which included synbiotic
banana puree 10% (fermented by Bif. lactis
BB-12 and Lactobacillus acidophilus),
especially with the stability of flavor criteria
(taste and smell). This is due to the
enhanced effectof the activity of probiotics
and the presence of banana puree.
Table 3. Sensory evaluation of ice cream treatments
Properties (Every
factor: 20 scores)
Treatments (After 1 day)
LSD value
Control Ice.1 Ice.2 Ice.3 Ice.4
Taste 15 17 18 17 18 2.47 *
Smell 14 15 18 16 18 2.55 *
Color 18 18 18 17 18 1.52 NS
Texture 14 13 12 16 16 2.07 *
Mouthfeel 13 13 14 16 16 2.41 *
Total (100%) 74% 76% 80% 82% 86% 5.91 *
Treatments (After 60 days)
Taste 15 16 18 16 18 2.08 *
Smell 13 15 18 16 18 2.17 *
Color 18 18 18 17 16 1.94 *
Texture 13 13 12 16 16 2.04 *
Mouthfeel 14 13 14 16 17 2.13 *
Total (100%) 73% 75% 80% 81% 85% 5.73 *
Treatments (After 120 days)
Taste 15 16 18 16 18 2.19 *
Smell 14 15 17 16 18 2.33 *
Color 18 18 18 17 17 1.25 NS
Texture 12 12 13 15 15 2.09 *
Mouthfeel 13 13 13 16 16 2.41 *
Total (100%) 72% 74% 79% 80% 84% 5.69 *
Quality level based on scores
1 to 4 5 to 8 9 to 12 13 to 16 17 to 20
Awful Bad Regular Good Excellent
* (P≤0.05).
* Statistically significant differences (P≤0.05), NS: Non-Significant statistical differences, Ice.1 and Ice.2 are
probiotic treatments including probiotic skim milk in the ratio 5% and 10%, respectively. Ice.3 and Ice.4 are
synbiotic treatments including banana pulp puree in the ratio 5% and 10%, respectively.
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
41
Ozturkoglu-Budak et al. (2019) indicated
that synbiotic fermented milk products
containing inulin with cultures of the
probiotics Lb. acidophilus La-5 and Bif.
animalis Bb-12 were preferred over
probiotic products by the sensory panel in
terms of appearance, body, and taste.
Regarding the color of the synbiotic ice
cream, the highest scores were in the trail
(Ice.4), reaching 18 on the first day, with a
slight fluctuation until the last day during
frozen storage, which is similar to the color
in the trail (Ice.3), which amounted to 17 on
the first day with a slight fluctuation until
the last day during frozen storage. This
difference is attributed to the high
percentage of fermented banana puree in the
trail (Ice.4), which amounted to 10% while
the trail (Ice.3) included 5% of fermented
banana puree.
As for the probiotic ice cream, the color
scores for both treatments (Ice.2 and Ice.1)
were close, reaching about 18 from the first
day until the last day during frozen storage,
because probiotic skim milk does not
include special-colored compounds. The
addition of 5% and 10% of fermented skim
milk in (Ice.1) and (Ice.2) successively
helped maintain the white color. The
panellists did not notice any significant
differences between the colors of the
probiotic ice cream samples in the first day
period and in the 120-day period. This is an
expected result in these samples because
they do not contain fruit pulp like those
found in synbiotic ice cream containing
fermented banana pulp. Significant
differences are observed between all the
samples under study on the 60th day,
because of continuing freezing storage,
which affects vitamin C and enzymes in the
banana fruit. This is justified the color
change in the ice cream containing the
fermented banana puree, as vitamin C
(ascorbic acid) is used to control enzymes in
the frozen fruit. Although the banana is not
heated before freezing, the oxidation of
ascorbic acid (vitamin C) occurs at a rate
slow during frozen storage. This leads to the
activation of the enzymes that cause the fruit
to turn brown especially with a loss of
vitamin C (Mieszczakowska-Frąc et al.,
2021). In treatment (Ice.4), the decrease in
color values increased, which led to
significant differences between all ice cream
samples generally. Banana contains
compounds that affect the color of ice
cream, especially Violaxanthin and
Cryptoxanthin, which are used as food
colorants. They are also one of the fruits
richest in trans-β-carotene compounds that
affect color properties (Sidhu and Zafar,
2018).
Regarding the texture trait of the
synbiotic ice cream, the texture scores for
both treatments (Ice.4 and Ice.3) were close,
reaching about 16 since the first day. Then,
they decreased slightly to 15/20 on the last
day during the freezing storage, and thus it
remained within the good range of the
texture properties, despite the different
added proportion of the fermented banana
puree as in treatment (Ice.4), which
amounted to 10% as compared to treatment
(Ice.3) which included 5% of fermented
banana puree. However, the panelists did
not notice a significant difference between
the two treatments regarding texture.
The results of our study agree with
Hasan et al. (2020), who indicated that the
use of ice cream samples containing 10% of
apple powder gave the highest values for
body and texture but decreased slightly with
an increase in the percentage due to the
roughness of texture and being granular.
They also mentioned that adding prickly
pear pulp by 5, 10 and 15% to ice cream
resulted in an improvement in body and
texture. In the same study, it was indicated
that the use of bananas in ice cream causes a
slight decrease in the texture properties of
the product as a result of an increase in the
total solids of the resulting ice cream, as the
banana powder is characterized by its high
content of carbohydrates. Yet. This differs
from our study where fermented banana
puree was used, not dried banana powder.
The presence of FOS reduces the firmness
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
42
of the product as it contributes to better
stability of the air bubbles embedded in the
ice cream and increases the production of
viscous gels, which leads to a better
acceptance of the texture properties
(Parussolo et al., 2017).
As for the probiotic ice cream, the
sensory evaluation scores for the texture
trait were lower than those of the synbiotic
ice cream. The texture scores for both
treatments (Ice.2 and Ice.1) were close,
reaching about 12-13 from the first day until
the last day during the freezing storage
period. Thus, they remained within the good
range of the texture properties, because of
using of 5% and 10% probiotic skim milk in
treatment (Ice.1) and treatment (Ice.2)
respectively which achieved an increase in
the percentage of non-fat solids, in
particular proteins, in the ice cream for both
treatments. As milk proteins have a
significant effect on the texture of ice cream
by reducing the size of ice crystals and
improving their stability when
polysaccharides are present (El-Aziz et al.,
2015). The results of our study for treatment
(Ice.2) agree Patel et al. (2006) who
indicated that increasing the proportion of
protein in the ice cream mixture leads to an
improvement in the texture properties as a
result of reducing the size of ice crystals.
Our study showed that treatment (Ice.2) at
the end of the freezing storage period
improved texture properties as compared to
the control.
Regarding the mouthfeel characteristic
of synbiotic ice cream, the mouthfeel scores
for both treatments (Ice.4 and Ice.3) were
close, reaching about 16 from the first day
to the last day during frozen storage. They
remained within the good range of
mouthfeel properties, despite the difference
in the percentage of fermented banana puree
added in the fourth treatment (Ice.4), which
amounted to 10% compared to treatment
(Ice.3) which included 5% of fermented
banana puree. However, the panelists did
not notice a significant difference between
the two treatments regarding mouthfeel, and
both treatments were acceptable. The results
of our study agree with Yangılar (2015) who
found that adding 1% and 2% banana pulp
flour to the ice cream mixture improved
mouthfeel trait equally for both treatments.
The addition of fermented bananas to ice
cream in treatments (Ice.3) and (Ice.3)
improved the texture and the mouthfeel
properties as compared to the control sample
because they have inulin, which gives a
distinctive viscosity due to the high
molecular weight of inulin. Inulin is also a
high hygroscopic substance and thus causes
water-binding. Therefore, it generates a gel-
like network which, together with other ice
cream ingredients, affects the rheological
properties of the product (Wood, 2011).
Inulin is characterized by poor solubility
depending on the length of its chain, and
therefore, when it comes into contact with
water or milk, it forms microcrystals,
generating a creamy gel material, and result
in a smooth, fat-like sensation in the mouth
(Villalva et al., 2017). Inulin also increases
the syneresis of probiotics towards the
production of more volatile fatty acids,
which in turn improves sensory properties
(Kamel et al., 2021). The results of our
study agree with Villalva et al. (2017) stated
that the addition of the probiotic
Bifidobacterium lactis Bb-12 with inulin in
peach ice cream at a concentration of 10%
did not result in negative effects on the
sensory properties of the product.
The sensory evaluation scores of the
mouthfeel sensation of the probiotic ice
cream were lower than those of the
synbiotic ice cream, the mouthfeel values
for both treatments (Ice.2 and Ice.1) were
close, averaging around 13-14 from the first
day until the end of the frozen storage
period. and thus remained within the good
range of mouthfeel properties because of the
use of 5% and 10% probiotic skim milk in
treatment (Ice.1) and treatment (Ice.2)
respectively increasing non-fat solids, in
particular proteins, in the ice cream.
Al-Sahlany and Al-musafer (2020)
indicated that a 10% replacement of wheat
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
43
flour with banana peel flour improved the
color and most rheological properties of
wheat flour samples after the replacement
process. Also, the viable cells of
Saccharomyces cerevisiae were
increased. Syed et al. (2018) indicated that
an increase in milk proteins leads to an
enhancement in the viscosity of ice cream
due to increased water binding and the
formation of smaller ice crystals. So the
product has a better mouthfeel, creaminess,
smoothness and the milkiness or opacity of
the product is improved. Thus, the synbiotic
ice cream scored mouthfeel points better
than the probiotic ice cream. The results
agree with Vacondio et al. (2013) who
reported that adding 5% and 10% yacon
extract to ice cream formulations gave
sensory acceptance level of 82.82% and
82.71%, respectively.
Overrun values of ice cream
Overrun values in table 4 showed the
control ice cream increased up to 71%, a
good value, as it is within the appropriate
range for the overrun in milk ice cream
which ranges between 50 to 85%. The value
indicates the correct homogenization of the
mixture, which facilitated mixing and the
incorporation of air with the mixture,
especially with milk proteins thus generated
suitable foam (Villalva et al., 2017).
The overrun values of probiotics ice
cream treatments were 73% and 77% for
treatment (Ice.1) and treatment (Ice.2),
respectively. Because the increase in total
solids including protein and carbohydrates
increase the ice cream overrun (Asres et al.,
2022). The overrun values of synbiotic ice
cream treatments were 75% and 78% for
treatment (Ice.3) and treatment (Ice.4),
respectively. As the use of fermented
banana puree by 5% for treatment (Ice.3)
and 10% for treatment (Ice.4) means an
increase in the percentage of total solids that
are different from the nature of the known
solids for ice cream. The reason may be due
to the fact that bananas contain oligofructose
and inulin which increase the incorporation
of air into the ice cream product. The ability
of insulin is greater than oligofructose in
this regard, which affects the ice cream
overrun (Wood, 2011).
The addition of inulin to the ice cream
mixture causes a large overrun in the
product, reaching 81%, i.e. it’s within the
standard in the Argentine legislation,
according to which the combined air should
not surpass 120% and the ideal overrun
value is from 70 to 100% (Villalva et al.,
2017). At this point, after achieving a good
percentage of overrun, a conviction was
generated that the proportion of banana
puree added to ice cream should not exceed
10% in order to avoid an increase in the
viscosity of the mixture, to achieve a
significant impact on the whipping rate (El-
Samahy et al., 2009). Also, studies have
also indicated that high levels of overrun
negatively affect the survival of probiotics
in ice cream, especiallywhen probiotics are
added to the mixture immediately before
freezing with constant stirring (Boza et al.,
2012).
An increase in the viscosity of the ice
cream mixture leads to an increase in the
overrun, depending on the nature and
percentage of the additive to the mixture.
However, high viscosity may reduce the
overrun of the product. Peasura et al. (2020)
stated that adding pumpkin by 25% gave the
overrun 73%, while it reached 84% in the
control sample. Elkot et al. (2022) reported
that the amount of black rice powder (BRP)
used in the manufacture of synbiotic ice
cream had a significant effect on the overrun
and concluded that increasing the
percentage of (BRP) led to increasing the
overrun to 41.80%. EL-Sayed et al. (2014)
reported that the using of prebiotic
Fructooligosaccharides at a rate of 2% with
different strains of probiotics in the
manufacture of synbiotic ice cream gave an
overrun range between 63.46 to 66.56%.
Mykhalevych et al. (2022) stated that the
addition of 1% oat beta-glucan to the ice
cream mixture led to an increase the overrun
to 72.3%. It seems that the fermentation
Diyala Agricultural Sciences Journal, 2023, Vol. (15) No. 1, 34-48
44
process has led to a change in the nature of
the banana puree, which supports the
achievement of the high overrun as shown
in Table 4 as a result of reducing the
viscosity to a degree. Srisuvor et al. (2013)
stated that prebiotics with the
exopolysaccharides (EPS) produced by
probiotic bacteria cause the casein particles
to be covered and the casein aggregates are
partially sterically stabilized, which leads to
a reduction in the effective volume fraction
and reducing viscosity.
Table 4. Overrun values of ice cream treatments
Overrun% Treatments
71.2±0.50 Control (only ice cream)
73.3±0.45 Ice.1 (Probiotic ice cream) (5%)
77.1±0.27 Ice.2 (Probiotic ice cream) (10%)
75.3±0.38 Ice.3 (Synbiotic ice cream) (5%)
Ice.4 (Synbiotic ice cream) (10%)78.2±0.82
Ice.1 and Ice.2 are probiotic treatments including probiotic skim milk in the ratio 5% and 10%, respectively.
Ice.3 and Ice.4 are synbiotic treatments including banana pulp puree in the ratio 5% and 10%, respectively
Conclusion
Possibility of using banana puree
fermented by Lactobacillus acidophilus
LA5 and Bifidobacterium lactis BB12 in
improving some properties of synbiotic ice
cream, included increasing viability of
probiotics, acceptable pH, improving the
sensory properties and additional to high
overrun. Banana puree leads to better
protection of probiotic cells during frozen
storage compared to the case of free
probiotic cells. Incorporation of probiotics
with banana puree in ice cream is possible to
obtain at least 10
7
viable cells/d.
Conflict of Interest
The authors declare that there are no
conflicts of interest regarding the
publication of this manuscript.
Acknowledgements
We would like to thank Department of
food science, College of Agricultural
Engineering Sciences, University of
Baghdad, Iraq. We would also like to extend
our deepest gratitude to all who have
contributed to the progress of this study.
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