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LWT - Food Science and Technology 91 (2018) 339–344

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LWT - Food Science and Technology


journal homepage: www.elsevier.com/locate/lwt

Development of non-dairy fermented probiotic drink based on germinated T


and ungerminated cereals and legume
Mayuri Chavana, Yogesh Gatb, Mugdha Harmalkara, Roji Waghmarea,∗
a
Food Science and Technology, School of Biotechnology and Bioinformatics, D. Y. Patil University, Navi Mumbai, India
b
Department of Food Technology and Nutrition, Lovely Professional University, Jalandhar, India

A R T I C L E I N F O A B S T R A C T

Keywords: In the present study probiotic drinks were developed using germinated and ungerminated seeds of barley, finger
Grains millet and moth bean. All three grains were washed, soaked, germinated, dried; both germinated and un-
Non-dairy milk germinated grains were roasted, grinded and mixed in the ratio of 2.5:1.5:1 (barley: finger millet: moth bean)
Lactobacillus acidophilus with sugar and cardamom. The drink mixtures were added to distilled water and milks like soy, almond and
Probiotic drink
coconut in different concentrations (0 g, 2 g, 4 g, 6 g and 8 g). Drink mixtures were inoculated with Lactobacilli
acidophilus, a probiotic bacterium. Physicochemical analysis like pH, acidity, bacterial enumeration and anti-
oxidant activity were carried out, along with the evaluation of organoleptic properties. Fermentation with L.
acidophilus starter culture improved the overall acceptability and functional properties of beverage during fer-
mentation. Changes in the pH and acidity, bacterial count, DPPH assay and polyphenol content were increased
as the concentration of drink mixture increased in three different milk and distilled water. In sensory evaluation
it was found that the coconut milk based probiotic drink scored highest than the water, soymilk, almond milk
probiotic drink. The overall acceptance score of 4 g drink mixture in probiotic drink was highest among all the
drink mixture concentration.

1. Introduction (Neus & Maite, 2015). Non-dairy probiotic drinks are available in
market which can be consumed by people having lactose intolerance,
Probiotic drinks can be made from variety of raw materials such as who cannot drink dairy-based probiotic drinks (Mradula & Somesh,
cereals, millets, legumes, fruits and vegetables (Mradula & Somesh, 2016; Vasudha & Mishra, 2013). This milk is a rich source of highly
2016; Rajyalakshmi et al., 2016; Vasudha & Mishra, 2013). Barley malt valuable proteins, iron, unsaturated fatty acids, dietary fibers, vitamin B
is a good source of starch, sucrose, vitamins, essential minerals and and isoflavones, which are vital part of our diet (He & Hekmat, 2014).
phytochemicals (Dabina-Bicka, Karklina, & Kruma, 2011). Cereal based Probiotics are live microorganisms which are introduced into host
probiotic products develop antimicrobial activity against common mi- for the health benefits they provide (Millette, Luquet, Ruiz, & Lacroix,
crobial pathogen (Sharma, Trivedi, & Gat, 2017). Germinated cereals 2008). Probiotic bacteria are used in the form of nutritional supple-
and grains are more nutritious than raw grains and cereals. Germinated ments and functional foods having advantage of easily digestibility. It
cereals are rich in digestible energy, bioavailable vitamins, minerals, also provides more soluble calcium, reduces flatulence, destroys un-
amino acids, proteins, and phytochemicals (Aboulfazli, Shori, & Baba, desirable pathogens and also improves taste and texture of probiotic
2016). Barely, ragi and moth bean help in management of various fermented food (Sharma et al., 2017). There are various microbial
diseases (heart diseases, diabetes, hyperlipidemia and obesity). strains which are used as probiotics. The most common bacteria used as
Functional beverages constitute one of the most developed segments probiotics on commercial basis are Bifidobacteria and lactic acid bac-
and are very appreciated for their nutritional characteristics in the teria (LAB) such as lactobacilli, lactococci and streptococci (Isolauri,
market (Aliakbarian et al., 2015; Silva, Bezerra, Santos, & Correia, Kirjavainen, & Salminen, 2002). L. acidophilus is a lactic acid bacteria
2015). Non-dairy milk is good alternative for dairy-based drinks be- which is rod shaped and gram positive in nature. Lactic acid fermen-
cause dairy milk contains whey and casein protein which can cause tation helps to increase nutritional quality of food as well as maintains
allergy. The advantage of non-dairy milk such as soy milk, almond milk the environment and shelf-life of the lactic acid bacteria (Mridula &
and coconut milk is due to the absence of cholesterol and lactose sugar Sharma, 2014).


Corresponding author. Food Science and Technology, School of Biotechnology & Bioinformatics, D. Y. Patil University, Sector 15, CBD Belapur, Navi Mumbai 400614, Maharashtra,
India.
E-mail addresses: roji.waghmare@dypatil.edu, rosewaghmare@gmail.com (R. Waghmare).

https://doi.org/10.1016/j.lwt.2018.01.070
Received 25 October 2017; Received in revised form 23 January 2018; Accepted 24 January 2018
0023-6438/ © 2018 Published by Elsevier Ltd.
M. Chavan et al. LWT - Food Science and Technology 91 (2018) 339–344

Various authors have carried out studies on non-dairy probiotic milk probiotic drink (GCPD) and ungerminated coconut milk probiotic
drinks. Mridula and Sharma (2014) prepared non-dairy probiotic drink drink (UGCPD) in different weights of 0 g, 2 g, 4 g, 6 g and 8 g. Hence,
using sprouted cereal, legumes and soymilk. Antioxidant activity and the concentration of drink mixture in probiotic drink was 0.02%,
polyphenol content in fermented soy milk supplemented with WPC-70 0.04%, 0.06% and 0.08%. Further, 4 g of sugar and 0.07 g cardamom
by probiotic Lactobacilli have also been previously studied (Subrota, were added to the drink mixture. The probiotic culture at a level of
Shilpa, Brij, Vandna, & Surajit, 2013). The mixture of geminated barley, 1ml/100 ml having cell count 104 cell/ml was added to the liquid
ragi and moth bean can thus be suitable for the growth of L. acidophilus portion containing the formulation mixture. Then the drinks were al-
as such mixtures may supply required amounts of carbohydrates, starch lowed to incubate for 6 h at 37 °C. All the samples were prepared in
and protein for growth of bacteria (Maselli & Hekmat, 2016). triplicates in a batch of 300 ml.
The objectives of the present work were (1) to develop cereal, grain
and legume based powder and increase its nutritional value, (2) to
2.5. Acidity
develop and evaluate non-dairy probiotic beverages using soymilk, al-
mond milk and coconut milk and (3) to compare the physicochemical
Acidity in terms of lactic acid was determined using titrimetric
properties of probiotic drink having germinated and ungerminated
method (Ough, Amerine, & Sparks, 1969). Titratable acidity was mea-
drink mixtures.
sured by titration against 0.1 N sodium hydroxide solution and using
1% ethanol solution of phenolphthalein as an indicator.
2. Materials and methods

2.1. Materials used 2.6. pH

Barley (Hordeumvulgare L.), ragi (Eleusinecoracana), moth bean pH of the non-dairy milk was measured using a pH meter
(Vignaaconitifolia), soyabean, almond and coconut used in this study (Equiptronics, India). The measurements were carried out in triplicates.
were procured from APMC market, Vashi, Navi Mumbai. Pure culture of
probiotic bacteria L. acidophilus was purchased from Jevan
2.7. Trolox equivalent antioxidant capacity (TEAC)
Pharmaceuticals, Navi Mumbai. All chemicals were supplied by
Himedia, Mumbai, India.
For antioxidant activity, probiotic drinks were diluted in 1:100 ratio
of probiotic drink: water. TEAC free radical scavenging activity was
2.2. Germination and milk preparation
calculated on the basis of the method described by Gat and
Ananthanarayan (2015). All the experiments were performed in tripli-
All the grains were cleaned, washed and soaked in water in the ratio
cates.
of 1:2 (seed to water) separately for 8 h at 30 °C. After 8 h of soaking,
water was drained and soaked grains were allowed to germinate in a
controlled germinator (30 °C and 95% relative humidity). Barely was 2.8. Total phenolic content
germinated for 48 h, ragi was germinated for 36 h while moth bean was
germinated for 24 h. The germinated grains were then dried in hot air Determination of polyphenols was determined using
oven at 55 ± 5 °C to 8% moisture content. Dried germinated and un- Folin–Ciocalteureagent (Gat & Ananthanarayan, 2015). Absorbance of
germinated grains were roasted for 5 min at 130 °C and ground in an the clear supernatant solution was measured at 765 nm using allic acid
electric grinder. Ground powder was sieved through 0.0075μ sieves. as a standard. Results were expressed as milligram gallic acid equiva-
For milk preparation, 250 g of soybean, 350 g of almond and 350 g lent per 100 g dry weight.
of coconut were soaked in water in the ratio of 1:6, 1:4.2 and 1:4 re-
spectively for 12 h at room temperature. After soaking, it were grinded
2.9. Sensory characteristics
it in an electric grinder and extracted milk was filtered through muslin
cloth. The milk was boiled for 5 min and cooled.
The sensory characteristics of probiotic drink samples were eval-
uated three times in three different sessions by a group of 15 panelists,
2.3. Preparation of culture
who had been trained following the standards procedure before con-
ducting the sensory evaluation (Kilcast, 2010; Waghmare & Annapure,
Probiotic culture was prepared by using lyophilized culture of
2015). 100 ml of probiotic drink sample in glass containers, duly coated
Lactobacillus acidophilus. Probiotic bacteria were streaked on MRS agar
with code was given to panelist for evaluating the sensory attributes
plate and for its preservation sub-culturing of bacteria was done after
like appearance, flavor, consistency, taste and overall acceptability
every 2 weeks. For preparation of inoculums, bacteria were inoculated
using nine point hedonic scale. Hedonic scale was in the following se-
in MRS broth and incubated for 48 h. After 48 h, broth was centrifuged
quence: like 9- like extremely, 8- like very much, 7- like moderately, 6-
at 8000 rpm and cell pellets were washed with saline water for 2–3
like slightly, 5- neither like nor dis like, 4- dislike slightly, 3- dislike
times until colourless pellet were observed. To obtain required cell
moderately, 2- dislike very much, 1- dislike extremely (Chen, Zhu,
count, optical density of inoculum was checked by spectrophotometer
Zhang, Niu, & Du, 2010).
at absorbance 660 nm.

2.4. Preparation of probiotic drink 2.10. Enumeration of probiotic count

All the germinated and ungerminated ground powder were mixed in Enumeration of viable cells was performed by estimating colony
the ratio 2.5:1.5:1 (barley: finger millet: moth bean). This ratio of drink forming unit number on De Man, Rogosa and Sharpe (MRS) agar plates
mixtures were added in 100 ml of milk and distilled water for the (pH- 6.4 ± 2) after incubation at 37 °C for 6 h. Probiotic count was
preparation of germinated distilled water probiotic drink (GDWPD), done by using streak plate method after incubation at 37 °C for 48 h.
ungerminated distilled water probiotic drink (UGDWPD), germinated Mixture of barley, ragi and moth bean is suitable for the growth of
soymilk probiotic drink (GSPD), ungerminated soymilk probiotic drink probiotic bacteria hence as there is increasing in concentration of drink
(UGSPD), germinated almond milk probiotic drink (GAPD), un- mixture there is increasing in count of probiotic bacteria (Maselli &
germinated almond milk probiotic drink (UGAPD), germinated coconut Hekmat, 2016).

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M. Chavan et al. LWT - Food Science and Technology 91 (2018) 339–344

Fig. 1. Effect of level of different concentrations of (a) germinated


and (b) ungerminated drink mixture in distill water probiotic
drink, soymilk probiotic drink, almond milk probiotic drink and
coconut milk probiotic drink on acidity (means of three re-
plicates ± standard deviation).

2.11. Statistical analysis drink and lowest in distilled water based probiotic drink. Increase in
supplement of nitrogen source resulted in a higher concentration of
All data were expressed as means ± standard errors of triplicate lactic acid therefore there is an increase in acidity of the probiotic drink
measurements and analyzed by SPSS for Windows (ver. 16.0). One-way samples (Kwon, Lee, Lee, Chang, & Chang, 2000).
analyses of variance (ANOVA) were carried out to test significant dif-
ferences (p ≤ 0.05). Mean and standard deviation were computed using 3.2. pH
Microsoft Excel 2010.
Changes in pH were determined after 6 h fermentation seen in
3. Results and discussion Fig. 2. pH of germinated distilled water, soymilk, almond milk and
coconut milk probiotic drink ranges from 5.84 to 5.33, 5.57 to 5.55,
3.1. Acidity 5.86 to 5.53 and 5.40 to 5.63 respectively. Whereas, the pH range of
ungerminated probiotic drink mixture is from 6.48 to 5.14, 5.94 to
Acidity of germinated and non-germinated probiotic drink is shown 5.91, 5.77 to 5.25 and 5.07 to 4.56 in distilled water, soymilk, almond
in Fig. 1. For both the germinated and ungerminated mixture probiotic milk and coconut milk probiotic drink respectively. The non-significant
drink, acidity increased significantly (P < 0.05) with the increase in difference (P > 0.05) was observed in all the germinated and un-
concentration of drink mixture in all the samples. Acidity of probiotic germinated probiotic drink samples. The non-significant change of pH
drink having germinated drink mixture ranges from 0.1 to 0.3, 1.03 to in probiotic drinks is due to hydrolysis of starch into sugars during
2.5, 1.8 to 3.5 and 0.3 to 1.5; while in case of probiotic drink mixture germination which is readily utilized by the organisms and converted to
having ungerminated drink mixture, acidity ranges from 0.1 to 0.7, lactic acid (Sharma et al., 2017).
0.27 to 1.43, 0.5 to 0.9 and 1.2 to 3 in distilled water, soymilk, almond
milk and coconut milk probiotic drink respectively. Acidity of drink 3.3. Trolox equivalent antioxidant capacity (TEAC)
mixture having germinated drink mixture is more as compared to un-
germinated drink mixture probiotic drink. Acidity was highest in al- The influence of level of different concentration of drink mixture
mond milk probiotic drink and lowest in distilled water based probiotic and different milk on trolox equivalent antioxidant capacity (TEAC)
drink germinated drink mixture. Whereas in non-germinated drink was evaluated (Table 1). Initially TEAC values of germinated distilled
mixture, highest acidity was observed in coconut milk based probiotic water, soymilk, almond milk and coconut milk probiotic drink were

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M. Chavan et al. LWT - Food Science and Technology 91 (2018) 339–344

Fig. 2. Effect of level of different concentrations of (a) germinated


and (b) ungerminated drink mixture in distill water probiotic
drink, soymilk probiotic drink, Almond milk probiotic drink and
coconut milk probiotic drink on pH (means of three replicates ±
standard deviation).

33.29, 0.70, 2.02 and 3.03 respectively. TEAC increased significantly 2.48 mM G.A.E and 1.40 to 2.13 mM G.A.E. respectively, in soymilk
(P < 0.05) for all the samples with the increase in quantity of germi- probiotic drink 2.02 to 3.2 mM G.A.E and 1.94 to 2.48 mM G.A.E, in
nated drink mixture in probiotic drink. Similarly for ungerminated almond milk probiotic drink 2.28 to 4.74 mM G.A.E and 2.22 to
probiotic drink values of TEAC increased significantly (P < 0.05) with 3.53 mM G.A.E and in coconut milk probiotic drink TPC ranged from
the increase in quantity of ungerminated drink mixture. TEAC of 3.10 to 4.77 and 3.12 to 4.20 mM G.A.E. TPC increased significantly
soyamilk probiotic drink was comparatively lower and increased line- (P < 0.05) with increasing amount of germinated and ungerminated
arly with increasing level of drink mixture concentration in probiotic drink mixture percentage in all the samples. For both the germinated
drink. Percentage scavenging activity of distilled water probiotic drink and ungerminated mixture, it was observed that the highest amounts of
was highest i.e. 33.2–50.7%, because the principle of DPPH assay states polyphenols are present in the probiotic drink containing of 8 g of drink
that any substance that when present at lower concentration compared mixture. Gallic acid equivalence was observed to be more in coconut
with those of the oxidizable substrate significantly delay or inhibit the milk probiotic drink and low in distilled water probiotic drink as
oxidation of that substrate. compared to other probiotic drink.

3.4. Total phenolic content 3.5. Sensory evaluation

Total phenolic content (TPC) in different probiotic drink with ger- The sensory evaluation was performed to found any sensorial dif-
minated and ungerminated drink mixture are presented in Table 1. ference between the distilled water, soymilk, almond milk and coconut
Total phenolic content in distilled water probiotic drink with germi- milk probiotic drink. It was also evaluated to see the difference ger-
nated and ungerminated drink mixture ranged from 1.44 to minated and ungerminated drink mixture powder in probiotic drink.

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M. Chavan et al. LWT - Food Science and Technology 91 (2018) 339–344

Table 1
Effect of level of different concentration of drink mixture and different milk on trolox equivalent antioxidant capacity (TEAC) and total phenolic content (TPC).

Drink mixture (g/100 mL) Distilled water probiotic drink Soymilk probiotic drink TEAC Almond milk probiotic drink Coconut milk probiotic drink
TEAC (% P.S.C) TEAC (% P.S.C) TEAC (% P.S.C) TEAC (% P.S.C)

Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated

a a a a a a a
0 33.29 ± 0.01 30.60 ± 0.02 0.70 ± 0.01 0.60 ± 0.1 2.02 ± 0.02 1.2 ± 0.02 3.03 ± 0.01 2.83 ± 0.3a
2 37.04 ± 0.02b 33.80 ± 0.01b 0.91 ± 0.01b 0.80 ± 0.01b 3.44 ± 0.01b 1.58 ± 0.02b 3.84 ± 0.01b 3.51 ± 0.02b
4 44.53 ± 0.01c 35.80 ± 0.03c 1.11 ± 0.02c 0.83 ± 0.02c 3.64 ± 0.1c 1.80 ± 0.1c 4.35 ± 0.02c 3.40 ± 0.02c
6 47.77 ± 0.03d 37.99 ± 0.11d 2.02 ± 0.01d 0.99 ± 0.03d 4.85 ± 0.01d 2.99 ± 0.04d 5.76 ± 0.03d 4.1 ± 0.01d
8 50.70 ± 0.1e 40.96 ± 0.1e 2.63 ± 0.01e 1.96 ± 0.02e 8.53 ± 0.03e 3.96 ± 0.01e 9.71 ± 0.02e 5.21 ± 0.01e

Drink mixture (g/100 mL) Distilled water probiotic drink Soymilk probiotic drink Almond milk probiotic drink Coconut milk probiotic drink
TPC (mM G.A.E) TPC (mM G.A.E) TPC (mM G.A.E) TPC (mM G.A.E)

Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated

a a a a a a a
0 1.44 ± 0.04 1.40 ± 0.01 2.02 ± 0.03 1.94 ± 0.04 2.28 ± 0.01 2.22 ± 0.02 3.10 ± 0.01 3.12 ± 0.03a
2 2.04 ± 0.01b 1.45 ± 0.02b 2.90 ± 0.02b 2.04 ± 0.01b 3.20 ± 0.03b 3.02 ± 0.01b 3.45 ± 0.02b 3.40 ± 0.02b
4 2.18 ± 0.03c 2.03 ± 0.01c 2.98 ± 0.01c 2.18 ± 0.03c 3.29 ± 0.02c 3.04 ± 0.1c 4.13 ± 0.01c 3.88 ± 0.01c
6 2.32 ± 0.01d 2.07 ± 0.01d 3.13 ± 0.01d 2.32 ± 0.01d 3.88 ± 0.02d 3.15 ± 0.01d 4.43 ± 0.01d 4.13 ± 0.01d
8 2.48 ± 0.02e 2.13 ± 0.3e 3.2 ± 0.01e 2.48 ± 0.02e 4.74 ± 0.0e 3.53 ± 0.03e 4.77 ± 0.3e 4.20 ± 0.01e

Where, P.S.C. - Percentage scavenging activity and mM G.A.E.- mille molar gallic acid equivalent. All values are mean ± standard deviation of three replicates. Different small letters
following the values in same column indicate differences for each level of drink mixture (P < 0.05).

Table 2 represent the appearance, flavor, consistency, taste and overall UGCPD received 7.1 score in visual appearance. In case of taste and
acceptability scores for all the probiotic drinks. A significant flavor, GCPD received 8.9 score and UGCPD received 8.2 score in flavor
(P < 0.05) difference in the selected sensory parameters was observed and 7.9 score in taste. GAPD received 7.7 and UGAPD received 7.4
among the different levels of drink mixture in the probiotic drink. In score in consistency. In conclusion, overall acceptability score of co-
appearance all the probiotic samples were acceptable. In this study, it conut milk based probiotic drink were highest as compared to distilled
was observed that the consistency of all the probiotic drinks having water, soymilk, almond milk probiotic drink samples. The score of all
concentration 4 g were good and were acceptable as a beverage. Sen- sensorial attributes of GCPD is highest in comparison to remaining
sory attributes of probiotic drinks having concentration 4 g received the probiotic drinks.
highest score by the panelists due to better appearance, flavor, con-
sistency, taste than remaining drink mixture concentrations. Therefore
probiotic drinks having 4 g of concentration were compared with the 3.6. Probiotic count
remaining drink mixture concentrations. In appearance, GSPD received
7.7 score; while UGSPD received 7.3. Also GCPD received 7.5 and Probiotic count of both germinated and ungerminated probiotic
drinks are shown in Fig. 3. The probiotic count for GDWPD were ranged

Table 2
Effect of level of different concentration of drink mixture and different milk on sensory attributes.

Sensorial Drink mixture, Distilled water probiotic drink Soymilk probiotic drink Almond milk probiotic drink Coconut milk probiotic drink
attribute g/100
Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated Germinated Non- germinated

a a a a a a a
Appearance 0 4.3 ± 0.10 4.3 ± 0.10 6.5 ± 0.22 6.5 ± 0.22 6.4 ± 0.11 6.4 ± 0.11 4.9 ± 0.13 4.9 ± 0.13a
2 4.6 ± 0.12b 4.2 ± 0.11a 6.6 ± 0.11a 6.3 ± 0.12a 5.2 ± 0.17b 5.1 ± 0.12b 6.1 ± 0.16b 5.8 ± 0.11b
4 5.4 ± 0.08c 5.1 ± 0.20b 7.7 ± 0.13b 7.3 ± 0.12b 7.2 ± 0.14c 7.1 ± 0.01c 7.5 ± 0.21c 7.1 ± 0.11c
6 5.1 ± 0.16d 4.9 ± 0.11b 6.7 ± 0.18c 6.5 ± 0.13c 7.1 ± 0.13c 6.9 ± 0.11c 6.6 ± 0.12d 6.4 ± 0.13d
8 5.1 ± 0.11d 4.5 ± 0.11c 6.2 ± 0.12d 6.0 ± 0.12d 6.5 ± 0.14d 6.2 ± 0.18d 6.7 ± 0.13d 6.4 ± 0.21d
Flavor 0 5.1 ± 0.22a 5.1 ± 0.22a 7.1 ± 0.13a 7.2 ± 0.13a 5.5 ± 0.14a 5.5 ± 0.14a 5.1 ± 0.21a 5.2 ± 0.13a
2 4.2 ± 0.16b 4.0 ± 0.20b 6.4 ± 0.11b 6.1 ± 0.17b 6.1 ± 0.13b 6.1 ± 0.13b 7.4 ± 0.18b 7.1 ± 0.19b
4 5.8 ± 0.21c 5.5 ± 0.11c 7.2 ± 0.23c 7.1 ± 0.22c 7.1 ± 0.17c 7.2 ± 0.11c 8.9 ± 0.12c 8.2 ± 0.21c
6 5.2 ± 0.12d 4.9 ± 0.13d 6.3 ± 0.16d 6.1 ± 0.13d 6.7 ± 0.11d 6.07 ± 0.13d 7.0 ± 0.13d 7.1 ± 0.21d
8 5.1 ± 0.13d 5.1 ± 0.15d 6.3 ± 0.22d 6.2 ± 0.21d 6.6 ± 0.13d 6.2 ± 0.11d 6.2 ± 0.2e 6.2 ± 0.24e
Consistency 0 4.8 ± 0.11a 4.8 ± 0.11a 6.3 ± 0.22a 6.3 ± 0.21a 5.6 ± 0.13a 5.6 ± 0.13a 5.1 ± 0.22a 5.2 ± 0.22a
2 4.2 ± 0.18b 4.1 ± 0.13b 6.1 ± 0.11a 6.1 ± 0.21a 6.7 ± 0.12b 6.2 ± 0.02b 6.1 ± 0.16b 6.1 ± 0.11b
4 5.8 ± 0.11c 5.6 ± 0.21c 7.2 ± 0.13b 7.0 ± 0.11b 7.7 ± 0.23c 7.4 ± 0.2c 6.7 ± 0.21c 6.3 ± 0.21b
6 5.2 ± 0.13d 5.1 ± 0.03d 6.2 ± 0.18c 6.02 ± 0.2c 7.3 ± 0.16d 7.1 ± 0.12c 6.1 ± 0.14d 6.0 ± 0.13b
8 5.2 ± 0.22d 5.1 ± 0.22d 6.5 ± 0.1d 6.2 ± 0.12c 6.8 ± 0.22e 6.4 ± 0.20d 6.5 ± 0.13e 6.1 ± 0.12b
Taste 0 3.2 ± 0.01a 3.2 ± 0.01a 6.7 ± 0.21a 6.7 ± 0.21a 5.7 ± 0.12a 5.7 ± 0.12a 6.9 ± 0.22a 6.9 ± 0.22a
2 3.5 ± 0.12b 3.2 ± 0.15a 7.0 ± 0.11a 6.9 ± 0.11a 6.1 ± 0.13b 6.0 ± 0.11b 7.8 ± 0.12b 7.1 ± 0.02a
4 5.6 ± 0.22c 5.3 ± 0.22b 7.8 ± 0.03b 7.5 ± 0.01b 6.7 ± 0.11c 6.6 ± 0.14c 8.9 ± 0.02c 7.9 ± 0.10b
6 5.5 ± 0.12c 5.2 ± 0.14b 6.1 ± 0.11c 6.2 ± 0.15c 6.9 ± 0.23c 6.5 ± 0.12c 7.7 ± 0.13d 7.2 ± 0.11c
8 5.2 ± 0.03d 5.1 ± 0.02b 6.7 ± 0.12d 6.3 ± 0.16c 5.1 ± 0.11d 5.0 ± 0.11d 6.7 ± 0.23e 6.3 ± 0.33d
Overall 0 4.1 ± 0.15a 4.1 ± 0.15a 6.4 ± 0.23a 6.4 ± 0.22a 5.7 ± 0.22a 5.7 ± 0.22a 5.6 ± 0.13a 5.6 ± 0.13a
acceptance 2 5.1 ± 0.15b 4.5 ± 0.11b 6.6 ± 0.1a 6.4 ± 0.12a 6.6 ± 0.16b 6.1 ± 0.13b 6.7 ± 0.10b 6.4 ± 0.11b
4 5.6 ± 0.12c 5.5 ± 0.13c 7.3 ± 0.15b 7.0 ± 0.05b 6.7 ± 0.21b 6.4 ± 0.21b 8.5 ± 0.23c 8.0 ± 0.12c
6 4.7 ± 0.13d 4.5 ± 0.01d 6.1 ± 0.18c 6.0 ± 0.11c 6.6 ± 0.10b 6.1 ± 0.11b 7 .1 ± 0.16d 6.9 ± 0.11d
8 4.3 ± 0.11e 4.2 ± 0.12e 5.8 ± 0.13d 5.1 ± 0.23d 6.4 ± 0.13b 6.2 ± 0.11b 7.6 ± 0.22e 7.5 ± 0.22e

All values are mean ± standard deviation of three replicates. Different small letters following the values in same column indicate differences for each level of drink mixture (P < 0.05).

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Fig. 3. Effect of level of different concentrations of (a) germinated and (b) ungerminated drink mixture in distill water probiotic drink, soymilk probiotic drink, Almond milk
probiotic drink and coconut milk probiotic drink on probiotic count (means of three replicates ± standard deviation).

from 8.1 to 8.60 log cfu/ml, for GSPD were 9.1–10.69 log cfu/ml, for Aliakbarian, B., Casale, M., Paini, M., Casazza, A. A., Lanteri, S., & Perego, P. (2015).
GAPD were 9.2–10.55 log cfu/ml and for GCMP were 9.47–11.07 log Production of a novel fermented milk fortified with natural antioxidants and its
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Acknowledgment cessing, storage and simulated gastrointestinal conditions. LWT - Food Science and
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Authors are grateful to Jeevan Pharmaceutical, Turbhe, Navi Subrota, H., Shilpa, V., Brij, S., Vandna, K., & Surajit, M. (2013). Antioxidative activity
and polyphenol content in fermented soy milk supplemented with WPC- 70 by pro-
Mumbai, India for providing the lyophilized culture. biotic Lactobacilli. International Food Research Journal, 20, 2125–2131.
Vasudha, S., & Mishra, H. N. (2013). Non-dairy probiotic beverages. International Food
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