formulation of guava fruit beverage with isolated

www.theinternationaljournal.org > RJSITM: Volume: 03, Number: 10, August-2014 Page 59

Formulation of Guava Fruit Beverage with Isolated Probiotic Strains

Sai Bramari.G1, Anitha.L

4* Kalpana.P

2 and Pooja Mandlik

3

1, 2, 3Research Scholars,

4Associate Professor

Department of Microbiology and Food Science & Technology, Institute of Science, GITAM

University, Visakhapatnam, A.P, India -530 045.

Email: [email protected]

ABSTRACT

The suitability of Guava fruit beverage for the production of probiotic juice with Lactic acid bacteria

isolated from milk, curd, whey and Lactobacillus plantarum has been studied. The probiotic strains

isolated from dairy products are characterized morphologically and biochemically. The formulated

Guava fruit beverage was inoculated with these isolates and incubated at 300C. Changes in pH,

acidity, sugar content, and viable cell counts during fermentation under controlled conditions were

measured. The pH of the Guava fruit beverage was initially 5.5 at 300 C and decreased gradually to a

suitable range. The titrable acidity at 300 C found to be 0.28% – 0.32% and was increased on further

fermentation. The sugar at 300 C was initially 15-22g/ 100ml. The viable cell counts at 30

0C were

estimated and they ranged between 5.6 x 10 6- 8.9x10

6. The fruit beverage was assessed for

acceptability through sensory evaluation. Molecular confirmation of isolate no.8 through 16s rDNA

sequence has been done and was found to be L. coryneformis. The final product resulted in suitable

pH, acidity, sugar content and ideal number of viable cell counts. The Probiotic Guava fruit beverage

could serve as a healthy beverage for consumers with dairy allergy, beneficial to gut health, prevention

of diarrhea and best nutrient source for under nourished vulnerable population.

Key words: Probiotics, Fermentation, Guava fruit beverage, Lactic acid bacteria, Diarrhea.

1. INTRODUCTION:

‘Let food be thy medicine and medicine be thy food’, the age-old quote by Hippocrates, is certainly the

tenet of today. With the growing interest in self-care and integrative medicine coupled with our health-

embracing baby boomer population, recognition of the link between diet and health has never been

stronger. Empirically, high levels of population, health go hand in hand with high levels of national

income. Higher incomes promote better health through improved nutrition, better access to safe water

and sanitation (Bloom and Canning, 2000).

Ever since the population is growing, the problems are also increasing very rapidly. The high

prevalence of malnutrition contributes to over 50% of child deaths. The second leading cause of death

among children under age five globally is Diarrhoea (Progress for Children Report, December 2007).

National surveys estimate that almost 10% of the population suffers from diarrheal disease in India.

(NFHS-3, 2005-06).

Diarrhea is mainly due to lack of adequate sanitation facilities for disposal of excreta, poor hygiene

practices which results in spread of pathogens in to the human environment (Progress for Children

Report ,December 2007; WHO, 2008). The current treatment for diarrhea includes the oral rehydration

therapy to replace the fluid and electrolyte loss. ORT (Oral Rehydration Therapy) was spread

worldwide to decrease the mortality rate due to diarrhea (Lochery, 2005). A search of alternative

therapy in treating various forms of diarrhea finally led to the conclusion that “Probiotics” can be the

treatment of choice (FAO/WHO, 2002). The studies demonstrated that the use of Probiotics reduced

the duration and the chances of symptoms lasting longer than four days by 60% (Sunil Kumar Mishra

et al., 2012). So, Probiotics are fast emerging as an alternative to conventional antimicrobial therapy

mailto:[email protected]


and are less likely to induce resistance due to their multifaceted mechanisms of action (Sujatha

Narayan et al., 2010).

Probiotics are defined as live microbial feed supplement that beneficially affects the host by improving

its intestinal balance (Gorbach, 2000). The term “Probiotics” meaning “for life” was first coined by

Lilly and Stillwell in 1960’s (Lilly and Stillwell, 1965). In the early 20th

century, Elie Metchnikoff

(1907) first proposed the concept of Probiotics. He suggested that beneficial bacteria could be

administered to replace harmful microbes with useful ones. FAO/ WHO (2002) defined Probiotics as

live microorganisms that when administered in adequate amounts confer a health benefit on the host

(FAO/WHO, 2002; Fuller, 1989). It has been estimated that 100 different species (with a total bacterial

population between 1010

and 1012

) are present in the human intestinal tract. Maintenance of an optimal

gut flora balance requires that ‘friendly’ bacteria, such as the gram-positive Lactobacilli and

Bifidobacterium which dominate and form a barrier to pathogenic bacteria. Probiotics are the most

natural and safe means of maintaining this balance (Anuradha and Rajeshwari, 2005).

Pribiotic dairy products and non dairy products like vegetable and fruits juices are the currently used

Probiotic foods (Vasudha and Mishra, 2011). The fermented milks and dairy products are the first

Probiotic products (Metchnikoff, 1907). Lactose intolerance and allergy towards dairy products are

problems with Probiotic dairy products (Amal Hassan et al., 2013). Hence it was thought that fruit

juices could be a better medium for fermentation with Probiotics (Panesar, 2011). The objective of this

research is to develop a Probiotic fruit beverage that contains Probiotic characters and healthy

properties of fruits.

2.0 Materials and Methods

2.1 Isolation of Lactic acid bacteria from dairy products

The Lactobacilli were isolated from different sources like raw milk, Curd, pasteurized milk, goat’s

milk and from whey (Renuka Goyal et al., 2012). MRS medium was used for the isolation of

Lactobacilli (Rogosa, et al, 1951). The raw milk of cow, curd of cow milk and whey were used as

inoculums and incubated at 370 C for 24-48 hours. (Talat Mehamood et al, 2009).

2.2 Morphological and Biochemical identification:

The phenotypic characterization of isolated strains was carried out by Grams staining (Talat

Mehamood et al, 2009) and Catalase test was also performed (James Cuppucino & Natalie Sherman,

2004). Along with isolated species of Lactobacilli, L. plantarum (NCIM NO. 2592) obtained from

NCIM, NCL; Pune was also tested to check its potentiality as a Probiotic strain. The isolated species

were found to be Gram positive and Catalase negative which gives a preliminary idea about the

isolated strains as Lactobacilli. These isolated strains were further subjected to the biochemical tests

like sugar fermentation tests, Arginine hydrolysis, Growth in different concentrations of NaCl and

growth at different temperatures.

After the confirmation of isolates through morphological and biochemical tests as Lactic acid bacteria

species, fermentation of Guava fruit beverage was carried out. (James Cuppucino & Natalie Sherman,

2004).

2.3 Formulation of Guava fruit beverage:

Fresh guavas were purchased from the market and were thoroughly washed. The skin was

removed and the fruits were made into pulp using kitchen mixer and juicer. Guava fruit beverage was

prepared as per composition (Table1) (Rakesh Gehlot et al. 2011).


TABLE: 1 Formulation of Guava Fruit Beverage

The concentrated Guava fruit beverage so prepared was diluted in 1:3 proportions and distributed into

clean, dry and sterilized glass bottles.

The isolated Lactobacilli were introduced into the diluted Guava fruit beverage and subjected

to fermentation by incubating the bottles at 300C (room temperature) and 4

0C (refrigeration

temperature). Shelf life of fermented Guava fruit beverage was checked at 300C and 4

0C. Fermentation

analysis of the Guava fruit beverage was carried out by checking its pH, titrable acidity, sugar

estimation and viable cell counts (Kyung Young Yoon et al., 2004).

2.4Fermentation analysis

2.4.1 pH:

The pH was measured with a pH meter (ELICO, sl.no. - 3456/2008 type 101).The pH was

measured at 300 C and 4

0 C.

2.4.2 Titrable acidity:

It was measured by titrimetric method. Initial and final readings of burette were noted. Titrable acidity

was calculated.

2.4.3. Sugar estimation:

The sugar content of fermented guava fruit beverage was estimated at frequent intervals by

phenol sulphuric acid method. It was estimated as grams of glucose per 100ml of guava fruit beverage.

O.D values were taken using spectrophotometer (Hitachi, u-2900) model.

2.4.4 Viable cell counts:

The viable cell counts were measured by the standard plate technique on MRS medium. The

inoculated and incubated Guava fruit beverage sample was taken and serially diluted. From 10 5

dilution, 0.1 ml of inoculum was taken and spread on to MRS agar plates and incubated for 24-48

hours. The colonies were counted using a colony counter. (Digital Colony Counter Sr. No- 1275)

(Kyung Young Yoon et al., 2005).

2.5 Antibacterial activity of isolates on Enterobacter aerogens, E.coli and S. aureus

The antibacterial activity by isolated samples on Enterobacter aerogens, Escherichia coli and

Staphylococcus aureus has been carried out by agar well diffusion method (Verma, et al., 2013).

2.6 Antibiotic sensitivity tests of isolates

The Antibiotic sensitivity of the isolated strains was carried out by disc diffusion method. Five oral

antibiotics like Ciprofloxacin, Ceftriaxone. Gentamycin, Lomefloxacin and Sublactum were used for

testing the isolated strains (Maharoshi Nayeem et al., 2012).

2.7 Molecular analysis:

One of the isolated strain (sample 8) was analyzed by 16s r DNA sequencing at Bioserve Biological,

India Lab at Hyderabad.

2.8 Statistical analysis:

All experiments were carried out in triplicates and each sample analyzed as duplicates. The

results were expressed as Mean ± S.D (Standard Deviation). Statistical analysis was done through

multiple comparison method (Scheffe’s post hoc test) of every isolate with each other. The P value

was analyzed by ANOVA (Kyung Young Yoon et al., 2004).

2.9 Organoleptic evaluation:

The sensory evaluation of fermented Guava fruit beverage was done by a selected panel

consisted of students from department of Food science who regularly participate in sensory analysis

studies and had experience in profiling food products. Sensory descriptors were developed during

GUAVA PULP 1LITRE

SUGAR 1.5 Kg

CITRIC ACID 22 Grams

WATER(FILTERED) 750ml.

SODIUM BENZOATE 2.0 Grams (Preservative)

RASPBERRY RED COLOUR 1 DROP


initial session by panelists. The score card consists of sensory attributes like appearance, color, flavor,

taste and overall acceptability. The scores given by panelists measured with five point hedonic rating

scale were recorded as mean± S.D (standard deviation) (Mohammad Daneshi et al., 2013).

3. Results

3.1 Morphology of isolates:

The present study was undertaken to study the suitability of Guava fruit beverage as substrate for

Lactic acid fermentation by Probiotic Lactobacillus strains. The experiment was started with the

isolation of Lactobacillus strains from dairy products like milk, curd and whey. As many as 50

colonies were isolated from each product but only 5 individual colonies were picked up after

screening. These isolated colonies were maintained as pure cultures. Out of these 5 isolates two were

isolated from raw milk of Cow, two strains from Curd of Cow milk and one isolate from whey. L.

plantarum (NCIM no. 2592) was obtained from NCIM, NCL, Pune. The isolated strains were

characterized morphologically. The isolated species were found to be Gram positive and Catalase

negative were shown in fig. 1 (James Cuppucino and Natalie Sherman, 2004). One isolate was

confirmed with molecular analysis of 16s rDNA sequencing (Isolate no. 8 shown in figure 4).

Fig 1: Morphology of Isolates of Lactobacillus from Diary Products (samples 2, 3, 8, 9 and 10

respectively)

3.2 Fermentation analysis

3.2.1 pH of the fermented Guava fruit beverage at 300C and 4

0C:

The pH of the Guava fruit beverage was initially 5.5 at 300 C and decreased gradually. (Table: 5) The

increase in the titrable acidity leads to decrease of the pH. The change in pH at 40C was shown in table

6. The same was represented graphically (fig. 2) which reveals that there was no significant difference

among pH values of each and every pair of sample at 300C. The decrease in the pH at 4

0 C was much

nearer to the changes in the pH at 300C. The similar results can be observed from the study of Kyung

Young Yoon et.al (2004).

TABLE 5: Changes in pH at 300 C during fermentation of Guava fruit beverage

Samples

0

days

3

days

5

days

7

days

9

days

12

days

Mean± S.D

F-

Value

P-

value

Decision

2 5.5 5.316 5.19 4.89 3.772 3.732 4.7333±0.7874 0.03 0.999 N.S

3 5.5 5.291 5.097 4.796 3.805 3.76 4.7100±0.7528

8 5.5 5.25 5.082 4.742 3.74 3.359 4.6117±0.8671

9 5.5 5.286 5.143 4.836 3.765 3.297 4.64±0.8949

10 5.5 5.321 5.168 4.739 3.83 3.402 4.66±0.8584

L.p 5.5 5.257 5.088 4.726 3.561 3.231 4.56±0.9436


TABLE 6: Changes in pH at 40C in fermented Guava fruit beverage

3.2.2 Titrable acidity (TA) at 300 C and at 4

0C in the fermented Guava fruit beverage:

The TA at 300 C in the Guava fruit beverage found initially to be 0.28% which was increased to 0.34%

for sample 2, 0.35% for sample 3, 0.37% for sample 8, 0.34% for sample 9, 0.36% for sample 10 and

0.38% for L. plantarum respectively. It was further increased and the same was shown in table 7 and in

the graph (fig. 2). TA at 40C was increased after fermentation (table 8) and was shown in the graph

(fig3) (Kyung Young Yoon et al. 2004).

TABLE 7: Variations in Titrable Acidity of Guava fruit beverage incubated at 300C

TABLE 8: Variations in Titrable acidity in fermented Guava fruit beverage at 40C

Samples

0 weeks

1stweek

2nd

week

3rd

week

4th

week

Mean± S.D

F -

Value

P-

value

Decision

2 5.5 5.384 5.182 4.983 4.741 5.156±0.3054 0.04 0.999 N.S

3 5.5 5.373 5.16 4.976 4.789 5.160±0.2868

8 5.5 5.356 5.107 4.945 4.654 5.114±0.3361

9 5.5 5.388 5.193 4.98 4.772 5.166±0.2974

10 5.5 5.379 5.204 4.963 4.734 5.154±0.3124

L. p 5.5 5.353 5.098 4.921 4.643 5.102±0.3415

Samples 0 days 3 days 5 days 7 days 9 days

12

days

Mean± S.D

F-

Value

P-

value

Decision

2 0.288 0.342 0.3708 0.3816 0.3888 0.3816 0.35833±0.03764 0.68 0.640 N.S

3 0.3024 0.3564 0.3924 0.3888 0.414 0.3888 0.37333±0.03933

8 0.3168 0.3708 0.396 0.4068 0.4188 0.414 0.38833±0.03764

9 0.288 0.3456 0.378 0.396 0.4032 0.4104 0.37167±0.04535

10 0.2952 0.3672 0.3924 0.396 0.3996 0.3996 0.37667±0.03933

L. p 0.3204 0.3852 0.4068 0.4188 0.4248 0.4161 0.39667±0.03933

Samples

0weeks

1stweek 2

ndweek 3

rdweek 4

thweek

Mean± S.D

F-

Value

P-

value

Decision

2 0.288 0.3924 0.432 0.522 0.5292 0.432±0.099 0.24 0.943 N.S

3 0.3024 0.3672 0.4476 0.5292 0.5328 0.436±0.100

8 0.3168 0.432 0.4896 0.5472 0.5508 0.468±0.0965

9 0.288 0.4032 0.4608 0.5328 0.5256 0.442±0.1008

10 0.2952 0.4176 0.4692 0.5292 0.5292 0.450±0.0956

L. p 0.3204 0.4896 0.4989 0.558 0.5652 0.488±0.1003

www.theinternationaljournal.org > RJSITM: Volume: 03, Number: 10, August-2014 4

TABLE 9: Changes in Sugar estimation of Guava fruit beverage incubated at 300C

Table10: Changes in Sugar estimation of fermented Guava fruit beverage at 40C

3.2.3 Sugar estimation (SE) of fermented Guava fruit beverage at 300 C and at 4

0C:

The SE value at 300 C was initially 15-22g/ 100ml for sample 2 to L.p respectively and has been

reduced during fermentation. The results of SE at 300C were tabulated in table 9 and at 4

0C in table 10

and were shown in the graph (fig. 2). The findings were on par with the study of Kyung Young Yoon

et al. (2005).

3.2.4 Viable cell counts of fermented Guava fruit beverage at 300C and at 4

0C in 10

5 dilutions:

The viable cell counts at 300C were estimated in 10

5 dilutions. The cell counts increased rapidly after

incubation of 24hours. They ranged between 5.6 x 10 6- 8.9x10

6 in 10

5 dilution. The cell counts were

found to be increased up to 7days after that these was a fall in the cell counts at 300C. This was shown

in the table 11 and fig.2. Viable cell counts at 40C found to be increased and were observed till

4weeks. The cell counts were shown in table 12 and fig.3. The increase in the cell counts and the

survival of culture in the fruit beverage makes it suitable as a probiotic fruit beverage. (Kyung Young

Yoon et al. (2005).

TABLE 11: Viable cell counts of isolates in Guava fruit beverage at 300C:

Samples 0

weeks

1st

week

2nd

week

3rd

week

4th

week

Mean± S.D F-

Value

P-

value

Decision

2 15.2 14.8 13.6 12.4 11.2 13.44±1.664 0.81 0.552 N.S

3 16.4 16.4 15.2 14 13.2 15.04±1.431

8 21.2 20.8 19.2 17.6 16 15.1±2.184

9 16.8 16.4 14.8 13.6 12.4 14.80±1.855

10 17.2 16.8 15.6 13.6 11.2 14.88±2.488

L. p 22 21.2 20 17.6 18 15.2±1.931

Samples 0

days

3

days

5

days

7

days

9

days

12

days

Mean± S.D F-

Value

P-

value

Decision

2 15.2 14.4 13.6 12.4 11.2 9.2 12.667±2.215 2.39 0.062 N.S

3 16.4 15.2 14 14 12.4 10.8 13.80±1.988

8 21.2 20 20.4 20.4 19.2 16.8 15.4±1.906

9 16.8 15.6 14 14.8 13.6 11.2 14.333±1.917

10 17.2 16.4 16 16.4 14.4 12 14.967±0.689

L.p 22 20.8 20.4 21.2 20 18 15.433±0.841

Samples 0 days 3 days 5 days 7 days

9 days

12 days

2 5.6x106 6.3x10

6 7.4x10

6 6.8x10

6 3.8x10

6 1.6x10

6

3 6.1x106 7.2x10

6 7.8x10

6 7.3x10

6 5x10

6 3.2x10

6

8 8.5x106 9.2x10

6 10.3x10

6 9.2x10

6 5.8x10

6 3.4x10

6

9 6.8x106 7.3x10

6 7.4x10

6 7.8x10

6 4.4x10

6 1.9x10

6

10 6.3x106 7x10

6 7.2x10

6 7x10

6 4.9x10

6 1.8x10

6

L. p 8.9x106 10x10

6 11.1x10

6 10x10

6 6x10

6 3.9x10

6


TABLE 12: Viable cell counts of isolates in Guava fruit beverage at 40C:

Fig. 2: Fermentation analysis at 30˚C; TA ---- Titrable acidity; SE ---- Sugar estimation

0

2

4

6

8

10

12

14

16

18

Ph at 30°C TA at 30°C SE at 30°C Viable Cell Count at

30°C

Parameters

Aver

age

Val

ue

Sample 2

Sample 3

Sample8

Sample 9

Sample10

Sample Lp

Samples 0 weeks 1stweek 2

nd week 3

rd week 4

th week

2 5.6x106 5.9x10

6 6x10

6 6.4x10

6 7x10

6

3 6.1x106 6.5x10

6 6.8x10

6 7x10

6 7.1x10

6

8 8.5x106 8.9x10

6 9x10

6 9.3x10

6 9.6x10

6

9 6.8x106 7x10

6 7.2x10

6 7.5x10

6 7.7x10

6

10 6.3x106 6.6x10

6 7x10

6 7.7x10

6 8x10

6

L. p 8.9x106 9.2x10

6 9.2x10

6 9.5x10

6 9.6x10

6

mple 2

ple 3


Fig 3: Fermentation analysis at 4˚C; TA ---- Titrable acidity; SE ---- Sugar estimation

3.2.5 Antibacterial activity:

The diameter of zone of inhibition was measured and recorded and shown in the table 13. The highest

zone of diameter was observed for L. plantarum followed by sample 8. All other strains have shown

specific antibacterial activity. Similar results can be observed from the work of Verma, et al. (2013).

3.2.6 Antibiotic sensitivity:

All the samples have shown resistance against the antibiotics like Ciprofloxacin, Ceftriaxone,

Lomefloxacine, Sublactum and Gentamycin. The results were shown in the table 14. The results of the

antibiotic sensitivity can be compared with the results of the study carried out by Maharoshi nayeem et

al. (2012).

3.2.7 Molecular analysis of isolate 8 through 16s r DNA sequencing:

The molecular analysis of sample 8 was carried out at Bioserve Biological India Labs,

Hyderabad. By the 16s r DNA sequence report the isolate was confirmed as L. coryniformis species.

The phylogeny of sequence was constructed using MEGA software (Fig 4). The sample 8 was chosen

for molecular analysis because it has shown highest antibacterial activity next to L. plantarum and the

sample 8 was shown resistance to all antibiotics. The molecular analysis was carried out similar to the

study of Mirzaei Hamid and Barzgari Abulfazl, (2012). Phylogenetic and molecular evolutionary

analyses were conducted using MEGA version 4 (Tamura et al, 2007).

Table 13: Anti Bacterial activity of isolates on Enterobacter aerogens , Escherichia coli and

Staphylococcus aureus:

Bacterial sp. Diameter of zone of

inhibition for E.

aerogens(mm)

Diameter of zone of

inhibition for E.

coli(mm)

Diameter of zone of

inhibition for S.

aureus(mm)

Sample-2 83 66 61

Sample-3 64 52 70

Sample-8 120 108 119

Sample-9 60 48 54

Sample-10 92 76 96

Sample-L. p 180 172 182

0

2

4

6

8

10

12

14

16

Ph at 4°C TA at 4°C SE at 4°C Viable cell Count at

4°C

Parameters

Av

erag

e V

alu

e

sample2

sample 3

sample8

sample9

sample 10

sample Lp


Table 14: Antibiotic sensitivity test of isolates on various antibiotics:

R- resistant S-sensitive

Fig 4: Evolutionary relationships of 8 taxa- Phylogenetic tree

4. Discussion:

The isolated colonies were identified as Lactobacillus species. L. plantarum was proved as best

probiotic strain in the earlier works (Kyung Young Yoon et al., 2004 & 2005). Sample 8 was analyzed

and identified through 16s rDNA sequencing as L. coryneformis. (Mirzaei Hamid and Barzgari

Abulfazl, 2012).

The Guava fruit beverage formulated, optimized and evaluated in the laboratory (Rakesh

Gehlot et al. 2011). The isolated five Lactobacillus species were inoculated into Guava fruit beverage

and fermentation was carried out at 300 C and 4

0 C.

All isolates used for fermentation of Guava fruit beverage were capable of utilizing the nutrients for

their growth, synthesis and metabolism. From the earlier works it can be known that the Lactobacillus

species were able to ferment vegetables such as Tomato, Cabbage (Kyung Young Yoon et al., 2004 &

2005) and fruits like Raspberries, Blue berries and Rock melon etc (Lin Kiat Saw et al., 2011). The

appearance, color, flavor, taste and overall acceptability of fermented Guava fruit beverage was found

be acceptable. The mean scores of the sensory attributes were given in the table 15. In fermented

Guava fruit beverage, the color was deep pink with creamy tinge and was accepted by 100%. The taste

was accepted by 99% and flavor was accepted by 95% of panelists. The sensory evaluation reports

were similar with the findings of Mohammad Daneshi et al. (2013). The fermented final product

resulted with desirable properties like suitable pH, acidity, the amount of reducing sugar and adequate

number of viable cell counts as in case probiotic Tomato, Cabbage and other fruit juices (Kyung

Young Yoon et al., 2004 & 2005). The isolated strains have shown significant antibacterial activity

against bacterial pathogens (Verma, et al., 2013). The isolated strains have shown good resistance

against antibiotics (Maharoshi Nayeem et al., 2012).

Sample

Ciprofloxacin

Ceftriaxone

Lomefloxacine

Sublactum

Gentamycin

2 R R R R S

3 R R S R R

8 R R R R R

9 S R R R R

10 R S R R R

L.P R R R R R


The richness of Guava fruit qualities and goodness of Probiotic strains blend together to make

Guava fruit beverage as an ideal Probiotic fruit beverage and acceptable to all vegetarians who are

allergic to dairy products.

TABLE 15: Mean scores of sensory attributes of Guava fruit beverage

CONCLUSION AND SIGNIFICANCE OF THE WORK:

.The Guava fruit is rich in vitamin C and is a good source of nutrients. The alkaline astringent

compound present in Guava fruit binds the bowels during diarrhea. So, the current research has been

undertaken to formulate and develop a Guava fruit beverage with isolated probiotic strains that

controls diarrhea and also provides the rich nutrients to consumers. The potentiality of the formulated

fruit beverage needs to be tested in vitro and in vivo to confirm and declare it as an anti diarrheal

probiotic fruit beverage.

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ITEM CODE APPEARANCE COLOUR FLAVOR TASTE OVER ALL

ACCEPTABILITY

Guava fruit

beverage at 30˚C

4.6±0.514

4.31±0.61

4.54±0.54

4.54±0.54

4.54±0.50

Guava fruit

beverage at 4˚C

4.4±0.66

4.6±0.43

4.5±0.54

4.65±0.39

4.35±0.39

ftp://ftp.fao.org/es/esn/food/


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formulation of guava fruit beverage with isolated

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