See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/371715464

Probiotic potential of Lactobacillus plantarum DMR14 for preserving and

extending shelf life of fruits and fruit juice

Article in Heliyon · June 2023

DOI: 10.1016/j.heliyon.2023.e17382

CITATION READS

1 104

12 authors, including:

Shirmin Islam Suvro Biswas

State University of Newyork at Binghumton University of Rajshahi
29 PUBLICATIONS 129 CITATIONS 26 PUBLICATIONS 421 CITATIONS

SEE PROFILE SEE PROFILE

Tabassum Jabin Md. Moniruzzaman

Bangladesh Reference Institute for Chemical Measurements University of Rajshahi
9 PUBLICATIONS 59 CITATIONS 8 PUBLICATIONS 19 CITATIONS

SEE PROFILE SEE PROFILE

All content following this page was uploaded by Md. Moniruzzaman on 20 June 2023.

The user has requested enhancement of the downloaded file.

 Heliyon 9 (2023) e17382

Contents lists available at ScienceDirect

Heliyon
journal homepage: www.cell.com/heliyon

Probiotic potential of Lactobacillus plantarum DMR14 for

preserving and extending shelf life of fruits and fruit juice
Shirmin Islam a, Suvro Biswas a, Tabassum Jabin a, Md. Moniruzzaman a,
Jui Biswas a, Md. Salah Uddin a, Md. Akhtar-E-Ekram a, Abdallah M. Elgorban b,
Gajanan Ghodake c, Asad Syed b, Md. Abu Saleh a, *, Shahriar Zaman a, **
a
Microbiology Laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi, 6205, Bangladesh
b
Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh, 11451, Saudi Arabia
c
Department of Biological and Environmental Science, Dongguk University-Seoul, Ilsandong-gu, Goyang-si, 10326, Gyeonggi-do, South Korea

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

Keywords: The harmful effects of chemical preservatives are driving the need for natural ones. To meet this
Lactic acid bacteria demand, probiotic lactic acid bacteria (LAB) were isolated from fermented oats in this study. The
Antimicrobial activity goals of this study were to separate and identify probiotic LAB from fermented oats, to determine
Probiotic potential
how effective these LAB are at combating pathogenic microorganisms in vitro, and to investigate
Preservation
Shelf life
their preservative capacity by applying the bacterium’s cell-free supernatant (CFS) to specific
fruits and fruit juice. The isolated strain was identified as Lactobacillus plantarum DMR14 using
morphological, biochemical, and molecular investigation. Antimicrobial, antibiofilm, anti-
oxidant, pH tolerance, and antibiotic resistance assays were used to evaluate the strain’s pro­
biotic potential, showing that Lactobacillus plantarum DMR14 had the strongest antagonistic and
anti-biofilm capacity against Shigella boydii. Furthermore, the bacteriocin-containing compounds,
cell-free supernatant (CFS) of the LAB, were tested against three fruits and one fruit juice, with the
cell-free supernatant (CFS) of the bacterium lengthening the shelf life of the fruits compared to
the untreated ones. Furthermore, while the concentration of coliform bacteria decreased in the
treated sugarcane juice, an increase in the concentration of lactic acid bacteria suggested that the
strain may be used as a fruit preservative in food industries.

1. Introduction

At present, a wide range of physical techniques and chemical reagents are employed for fruit and fruit juice preservation processes.
These preservation methods aim to prevent or delay the spoilage of fruits and fruit juices caused by microbial activity, oxidation, or
enzymatic degradation. Physical techniques, such as refrigeration, freezing, and dehydration, are frequently used to prolong the shelf
life of fruits and juices. Chemical reagents, such as antioxidants, preservatives, and antimicrobial agents, are also commonly used to
preserve fruits and juices. However, the use of these methods may have drawbacks, such as changes in the nutritional and sensory
qualities of the preserved products [1,2]. Apart from their ability to regulate unwanted microorganisms, traditional techniques such as
filtration or thermal processing may also negatively impact the natural components of biological products, such as aromatic

* Corresponding author.
** Corresponding author.
E-mail addresses: saleh@ru.ac.bd (Md.A. Saleh), szaman@ru.ac.bd (S. Zaman).

https://doi.org/10.1016/j.heliyon.2023.e17382
Received 4 February 2023; Received in revised form 9 June 2023; Accepted 15 June 2023
Available online 19 June 2023
2405-8440/© 2023 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
S. Islam et al. Heliyon 9 (2023) e17382

compounds, flavoring ingredients, and other compounds. This means that while these methods are effective in preserving food, they
may alter its overall quality and flavor [3]. There are several chemical substitutes that can be used as food preservatives, and among
them, the most commonly used ones are chlorine-based compounds [4]. However, alternative methods have also been developed to
achieve similar results. For instance, organic acids, ozone, peroxyacetic acid, electrolyzed water, hydrogen peroxide, and
calcium-based solutions [5] have been found to be effective in preserving food and preventing spoilage. These substitutes offer diverse
options for food manufacturers to choose from, depending on the specific needs and requirements of their products. Consumer concern
over food safety has generally increased due to the widespread misuse of chemical synthetic food preservatives, which has prompted a
search for natural and safe food preservatives to replace the chemical synthetic ones [6]. Natural antibacterial agents may be used
effectively to preserve food and maintain its quality and safety. Consumers are progressively becoming more interested [7] in the
innovative and alternative techniques of food bio preservation [8]. Fresh-cut fruits and vegetables have a very short shelf life and
experience severe physiological deterioration, and biochemical changes and microbiological degradation may occur while selling. All
of these changes can cause considerable loss of sensory attributes, such as color, smell, and flavor, as well as the development of
harmful and dangerous organisms, resulting in a decrease in shelf life [7,9]. Biopreservation is a natural and regulated process that can
help ensure the safety of fresh fruits and vegetables, making them a healthier and more sustainable option for consumers. Fresh fruits
and vegetables, including those that have been freshly cut, can be preserved using biopreservation methods, which extend their shelf
life by utilizing natural and safe microflora without any harmful biologically active substances. This technique involves introducing
beneficial bacteria, such as lactic acid bacteria or other probiotics that can prevent the growth of harmful microorganisms while also
maintaining the quality and freshness of the produce [10,11]. Probiotics, which are living bacterial species that are beneficial to
human health, are being utilized in the food industry as preservatives to prevent the growth of harmful microorganisms and as po­
tential substitutes for antibiotics to prevent or treat various diseases [12]. The majority of probiotics used in food products belong to
the group of lactic acid bacteria (LAB), which includes Bifidobacterium and Lactobacillus. These bacteria are capable of producing
several active metabolites, such as bacteriocins, reuterin, and hydrogen peroxide, which are low molecular mass compounds that can
effectively combat harmful microbes. Moreover, these active compounds produced by probiotics are safe, natural, and have no adverse
effects on human health, making them a healthier and more sustainable alternative to traditional preservatives and antibiotics [13,14].
In addition to their ability to produce active metabolites that combat harmful microorganisms, LAB is also classified as “Generally
Recognized As Safe” (GRAS) by the US Food and Drug Administration (FDA) due to their immunomodulatory, antibacterial, anti-­
biofilm, and antioxidant properties. These beneficial properties of LAB make them an attractive alternative to synthetic food pre­
servatives and antibiotics, as they offer a natural and safe solution for food preservation and promoting human health. Furthermore,
the GRAS status of LAB highlights their widespread acceptance and use in the food industry, indicating their potential for future
development and applications in food technology [15].
Lactic acid bacteria can be obtained from various sources, but dairy products are considered the most viable and suitable source for
probiotics that offer potential health benefits to consumers [16,17]. Despite this, the high sugar content, cholesterol levels, and lactose
intolerance issues associated with dairy products have hindered the development of dairy-based probiotic products. Therefore,
alternative sources of lactic acid bacteria are being explored, such as plant-based sources like fermented vegetables, fruits, and grains.
These non-dairy sources offer a more sustainable and healthier option for probiotic food products, which can cater to a wider range of
consumers with different dietary restrictions or preferences [18,19]. Table 1 depicts several cereal-based sources of lactic acid bacteria
from previous studies, indicating that cereal-based foods may be an enriched source of probiotic lactic acid bacteria.
To explore alternative sources of lactic acid bacteria for use as probiotics and natural preservatives, the study aimed to isolate new
probiotics from one of the most commonly consumed cereal-based foods. In addition to identifying a new source of lactic acid bacteria,
the study aimed to assess the probiotic potential of the isolated strain and its ability to naturally preserve fruits and fruit juices, offering
a more sustainable and eco-friendly preservation technique. The study’s findings suggest that the isolated strain has the potential to be
used as a probiotic and natural preservative, providing a promising alternative to conventional preservatives that are associated with
adverse health and environmental effects. By utilizing natural and safe microflora, this approach could promote healthier and more
sustainable food preservation practices.

Table 1
Cereal-based foods sources of lactic acid bacteria.
Cereal based foods Name of the lactic acid bacteria References

Rice Lactobacillus brevis, Lactobacillu curvatus, Lactobacillus plantarum DMR14, Lactobacillus farciminis D323 [20–22]
Maize Lactobacillu fermentum, Lactobacillu rhamnosus, Lactococcus, Pediococcus [23–25]
Sorghum Lactobacillus. delbrueckii, Lactobacillu, fermentum [26]
Wheat Lactobacillus coryniformis, Leuconostoc mesenteroides, Leuconostoc mesenteroides, Lactobacillus brevis, Lactobacillus. Acidophilus [27–29]
Rye Weissella paramesenteroides, Lactobacillus, Lactobacillus. Kimchi, Lactobacillus plantarum DMR14, Lactobacillus brevis [30,31]
Barley Lactobacillus spp. Pediococcus acidilactici, Enterococcus faecalis, Lactobacillus plantarum DMR14 [32]
Oat Lactobacillus plantarum DMR14, Streptococcus thermophilus DSM 20259, Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 [33,34]
Chia Lc. Lactis, Lactobacillus plantarum DMR14 [35]
Millet Lactobacillu. fermentum, , Lactobacillus spp. [36,37]

2
S. Islam et al. Heliyon 9 (2023) e17382

2. Materials and method

2.1. Sample collection and isolation of the bacteria

Quaker oats samples were collected from a super shop of Shaheb-bazar Rajshahi, Bangladesh, and brought to the microbiology
laboratory, department of Genetic Engineering and Biotechnology, University of Rajshahi, where 5 g of oats sample was fermented
overnight.
LAB were isolated on MRS agar after incubation with MRS broth for 12 h at 37 ◦ C. Plates were then incubated at 37 ◦ C for 24 h
aerobically. An isolated strain was sub-cultured in MRS broth. Glycerol stocks (50% v/v) were prepared for each colony and stored at
− 80 ◦ C.

2.2. Molecular analysis

According to the procedure described by Naeem et al. [38], the genomic DNA of the bacteria in its pure form was isolated. To
achieve this, universal forward and reverse primers, specifically 1492R (5′ - GGTTACCTTGTTACGACTT-3′ ) and 27F
(5′ -AGAGTTTGATCCTGGCTCAG-3′ ), were employed. The amplified products from the PCR process were then submitted for 16S
rRNA-sequencing to a commercial sequencing service (Invent Technologies Ltd. Dhaka, Bangladesh).

2.3. Phylogenetic analysis

Using BLAST and the Gene Bank Internet service, it was possible to identify the bacterial strains down to the species level. The 16S
rRNA sequence data was then uploaded to the GenBank database (https://submit.ncbi.nlm.nih.gov) and MEGA-X software was used to
carry out the phylogenetic and molecular evolutionary study.

2.4. Probiotic properties

The chosen bacterial strain underwent additional screening to determine its probiotic qualities. The LAB species was employed, and
the probiotic qualities of the species were carefully examined. The bacterial strain needs to successfully complete each of the tests
required at this stage, in accordance with FAO/WHO recommendations, in order to establish its viability as a possible probiotic
candidate (https://www.fao.org/3/a0512e/a0512e.pdf). This screening employs a number of in vitro tests, which are described in
more details in the following sections.

2.4.1. Acid tolerance

Acid tolerance of the chosen bacterial strain was investigated. The medium used was MRS broth containing 3 mg/mL of pepsin
enzyme. 1.0 N HCl was used to adjust the pH of broth at various pH values (2.0, 3.0, 4.0, 5.0, 6.0, 7.0, and 8.0), and the same solution
was used as a control set (pH 7.0) [39]. The broth was infused with overnight-grown cultures of potential strains and was then
incubated for 24 h at 37 ◦ C. The sampling process was performed every 12 h. At 620 nm, the optical density was determined, and viable
counts were also taken into account.

2.4.2. Bile salt tolerance

This test was performed using oxgall (HiMedia) at a concentration of 0.3% (w/v)). The isolated overnight cultured strain was then
spread on it and the presence and absence of the bacteria was observed.

2.4.3. Antibiotic susceptibility (Kirby-Bauer) test

The technique employed to obtain information on the resistance of bacterial isolates to antibiotics was the Kirby-Bauer disc
diffusion method with some modifications [40]. This technique categorized the obtained results as either susceptible, intermediate
resistance or resistant. The interpretation of the zone diameters was done using the performance standards for antimicrobial disk
susceptibility tests.

2.4.4. Antioxidant capacity of intact cells

2.4.4.1. Preparation of intact cells. To obtain intact cells, a 24-h broth culture was prepared and then centrifuged at 10,000 rpm for 10
min. The resulting cell density was adjusted to 108 CFU/mL using MacFarland 0.5. This adjustment was made using 0.85% saline.

2.4.4.2. DPPH test. The DPPH free radical method was used to observe the antioxidant activity of the chosen strain. This method was
modified according to the description provided by Yang et al. [41. To summarize, the experiment involved mixing 2 mL of whole cells
with 1 mL of methanolic DPPH solution and incubating them for 30 min in the absence of light. After this, the solutions were subjected
to centrifugation at 6000 rpm for 10 min. Anhydrous methanol was used as a blank, while a mixture of 2 mL of methanol and 1 mL of
DPPH was employed as a control. Optical density measurements were taken at 517 nm, and the scavenging activity was calculated
using a specific formula. The GENSYS 10S UV-VIS Thermo Scientific Spectrophotometer was utilized to determine the OD value. The

3
S. Islam et al. Heliyon 9 (2023) e17382

following equation was used to calculate antioxidant percentages:

Scavenging rate (%) = 1 − (sample − blank) control × 100%

2.5. Screening of lipolytic activity

Tween 80 media (Oxoid Ltd., Basingstoke Hampshire, UK) was used for the evaluation of lipase activity according to established
protocol [41]. Briefly, tween 80 was added to TSA at a ratio of 1:100. Then, phenol red (Sigma-Aldrich, St. Louis, MO, USA) was added
as an indicator. The LAB was then streaked on the plate and incubated for 24–48 h at 37 ◦ C. Color change from red to yellow–orange
indicated positive results.

2.6. Screening of amylolytic activity

2.5 g of starch agar (Himedia, India) were added to 100 mL distilled water and autoclaved at 121 ◦ C for 15 min. The probiotic strain
was then streaked on the media and incubated for 24 h at 37 ◦ C. Then, the media was flooded with 1% iodine solution (Himedia, India)
[41].

2.7. Screening of hemolytic activity

Overnight cultured LAB was grown and streaked on a blood agar plate (4% sheep) (HiMedia Laboratories, Mumbai, India) and
incubated for 48 h at 37 ◦ C. The presence or absence of zones of hydrolysis around the colonies was examined [42].

2.8. Antimicrobial tests

The antibacterial activity of the strain of lactic acid bacteria was evaluated through the agar-well diffusion method, which was
carried out based on prior literature, with certain adjustments [43]. The strain was examined for its ability to generate antimicrobial
substances against the bacteria that were tested (Table S2). Mueller Hinton agar plates were produced and the indicator bacteria were
added. Wells with a diameter of 5 mm were created in the agar plates and loaded with 100 μl of the tested strain’s culture. The plates
were then incubated at 37 ◦ C for 24–48 h. The diameter of the clear zone of inhibition was measured in millimeters.

2.9. Antibiofilm tests

2.9.1. Biofilm formation assay

Biofilm formation test was performed following the protocol ofmicrotiter plate (MtP) assay [44]. Here, the selected bacterial
strains (100 μl of each strain) (Table S2) were allowed to grow in wells of a 96-well microtiter plate (Tarsons, India) containing 100 μl
of luria bertani (LB) liquid medium and then incubated on an incubator at 37 ◦ C for 24 h without shaking to allow the formation of
biofilm at the bottom of the wells. After the incubation period, the plate was rinsed twice with double distilled water, air-dried, and
then oven-dried at 37 ◦ C for 1 h (60 min). The biofilm was fixed with 150 μl of absolute methanol for 5 min. After washing, the biofilm
was stained with crystal violet (0.1%) for 15 min. Finally, 100 μl of dilution was quantified by measuring at 595 nm.

2.9.2. Inhibition of biofilm formation by cell free supernatants (CFSs)

The previously described protocol (2.9.1) was used to assay the biofilm inhibition ability of the cell free supernatants with some
modifications. Here the effect of cell free supernatants (CFSs) of the bacteria on biofilm formation was measured using the co-
incubation method. Here, 100 μl CFSs (centrifuged at 12000 rpm for 20 min at 4 ◦ C to obtain cell free supernatants) was added to
the bacterial inoculum. Disruption percentage was calculated using the following equation:
Disruption Percentage = (OD control – OD sample) × 100 / control

2.10. Extending and preserving shelf life of fruit and fruit juice with LAB

2.10.1. Preservation of fruits

The approach proposed by Dhundale et al. was used to investigate the use of LAB in prolonging and maintaining the freshness of
fruits [45] with some modifications. The culture was added to 100 mL of MRS broth, which was then incubated for 48 h at 37 ◦ C. After
incubation, the broth was centrifuged at 10,000 rpm for 20 min and the resulting supernatant was used as a biofilm coating agent. The
fruits in the experimental group were coated with the supernatant after being dipped in it, while the control group consisted of un­
coated fruits dipped in sterile distilled water. The coated fruits were dried with air for an hour to eliminate any surface moisture.

2.10.2. Preservation of fruit juice

The locally available sugarcane juice was used to preserve it. The lactic acid bacterium was grown on MRS broth and incubated it at
37 ◦ C for 24 h. After centrifuging at 4000 rpm for 10 min, a 15 mL of CFS was collected. Then, we inoculated 1 mL of CFS into 20 mL of
sugarcane juice and incubated it at 37 ◦ C for 48 h. Sampling was performed every 24 h for microbiological and chemical analysis.

4
S. Islam et al. Heliyon 9 (2023) e17382

Viable cells were counted using the standard plate count method with MRS agar and Mac-Conkey agar medium and expressed the
results as colony-forming units per milliliter of sample (CFU/mL).

2.11. Statistical analysis

Each test was analyzed in duplicate, and the entire experiment was run in triplicate. The outcomes are presented as mean SD
(standard deviation). The statistical computer program SPSS 13.0 was used to examine the experimental data (Apache Software
Foundation, USA).

3. Results

3.1. Bacterial isolation

In this study, the LAB was initially cultured on Selective De Man, Rogosa, and Sharpe (MRS) media. The process was maintained in
triplicate to ensure consistency. The selection of a pure isolate of the LAB strain was based on observing the colony characteristics of
the plate (Fig. S1).

3.2. Bacterial identification based upon 16S rRNA sequencing

After the completion of the 16SRrNA gene sequencing, the obtained sequence was compared with the 16S rRNA gene sequences of
various organisms present in the NCBI Genebank database. Where, numerous sequences were discovered that exhibited significant
similarity to the isolate. Among them, the isolate demonstrated a higher degree of identity (92.24%) to Lactobacillus plantarum DMR14.

3.3. Phylogenetic position of bacterial isolate

Relationships of the bacterial strain Lactobacillus plantarum DMR14 to similar isolates were deciphered using a phylogenetic tree
(Fig. 1).

3.4. Probiotic properties of the Lactobacillus plantarum DMR14

3.4.1. pH tolerance test

Acid tolerance of the bacteria was observed at various pH levels ranging from 2 to 7. The optimum pH of the bacteria was 6.0
(Fig. 2).

3.4.2. Bile salt tolerance

In the present study, Lactobacillus plantarum DMR14 was able to grow in the presence of bile salts (Table 2). Tolerance to bile salts is
required for the growth and metabolic activities of bacteria in the intestine of the host.

3.5. Antibiotic susceptibility test

An antibiotic sensitivity test was done for the selected strain, with the results shown in Table 3. The strain showed resistance against
amoxicillin (AMX) and erythromycin (E), as well as susceptibility to kanamycin (K), gentamycin (G), doxycycline (DO), and chlor­
amphenicol (C).

Fig. 1. Phylogenetic tree showing the evolutionary relationship of the isolated bacterial strain. A maximum likelihood nearest-neighbor interchange
tree was constructed using MEGA 11.

5
S. Islam et al. Heliyon 9 (2023) e17382

Fig. 2. pH tolerance level of the Lactobacillus plantarum DMR14. Various pH ranging from 2 to 7 were used. Optical density was measured at 600
nm. Data were recorded after 24 h of incubation.

Table 2
Bile salt (0.3%) results of Lactobacillus plantarum DMR14.0.3% bile salt was used for this study. Data were recorded 4 different time durations.
Name of the Bacteria Bile Result (3 h) Bile Result (6 h) Bile Result (9 h) Bile Result (12 h)

Lactobacillus plantarum DMR14 Positive Positive Positive Positive

Table 3
Antibiotic sensitivity test result. 9 different antibiotics were used. Here, S indicates sensitive, I indicates
intermediate resistant, and R indicates resistant.
Antibiotics Name Zone of Inhibition (mm) Status

Kanamycin (K) 20.33 ± 1.52 S

Gentamycin (G) 19.67 ± 2.08 S
Penicillin (P) 11 ± 1 I
Ampicillin (AMP) 10 ± 1 I
Ciprofloxacin (CIP) 28.33 ± 2.88 S
Amoxicillin (AMX) 6 R
Doxycycline (DO) 18 ± 1 S
Chloramphenicol (C) 21 ± 1 S
Erythromycin (E) 6 R

Note:Resistant=<10 mm; Intermediate = 10–15 mm; Susceptible=>15 mm.

3.6. Antioxidant activity test

An antioxidant test was carried out on the isolate. The result indicated that the strain had 43.6% DPPH scavenging activity
(Table 4).

3.7. Amylolytic, hemolytic, and lipolytic activity tests results

The selected putative probiotic bacterial strain was tested using various in vitro enzymatic potential tests. In this study, Lacto­
bacillus plantarum DMR14 showed positive lipolytic, proteolytic, and amylolytic activities (Table 5).

3.8. Antagonistic test

Lactobacillus plantarum DMR14 displayed varying degrees of antimicrobial activity against six pathogenic bacteria. The bacterium’s
cell-free supernatant (CFS) effectively suppressed biofilm formation in all bacteria tested. Notably, the highest inhibition rate (67.7%)

Table 4
Antioxidant activity test result of the intact cells of Lactobacillus plantarum
DMR14.
Bacterial Name Antioxidant Percentage

Lactobacillus plantarum DMR14. 43.6%

6
S. Islam et al. Heliyon 9 (2023) e17382

Table 5
Lipolytic, amylolytic, and hemolytic activity test results
of Lactobacillus plantarum DMR14.
Test Name Result

Amylolytic Positive
Hemolytic Gama hemolytic
Lipolytic Positive

was observed against Shigella boydii. Relatively weaker inhibition activity was observed against Pseudomonas sp., Aeromonas sp., and
Staphylococcus aureus, while no significant inhibition was observed against Escherichia coli and Bacillus cereus (Table 6).

3.9. Antibiofilm test

3.9.1. Biofilm formation assay

Biofilm formation is one of the main causes of antibiotic resistance. All bacteria showed strong biofilm formation ability. The
biofilm formation abilities of the selected six bacterial strains are shown in Table 7.

3.9.2. Biofilm inhibition of the LAB CFS

The findings revealed a significant reduction in biofilm formation following treatment with the LAB’s cell-free supernatants (CFSs).
Among the tested strains, the lowest anti-biofilm activity (25.64%) was observed against Staphylococcus aureus. Notably, the attach­
ment of Shigella boydii to the microtiter plates was markedly inhibited, with a reduction of 67.87%. The biofilm inhibitory potential of
the CFSs against bacterial pathogens is visually presented in Fig. 3.

3.10. Extending and preserving the shelf life of fruit and fruit juice with Lactobacillus plantarum DMR14

The aim of this study was to investigate the impact of LAB on the shelf life of selected fruits when stored under laboratory conditions
at room temperature (approximately 37 ◦ C). To assess the extension of fruit shelf life, the cell-free supernatants (CFSs) of Lactobacillus
plantarum DMR14 were applied to the fruits. Throughout a 7-day observation period, whole apples treated with CFSs remained fresh
and free from contamination (Fig. 4 A, B, and C). Similarly, in the case of chopped apples, the treated samples exhibited better
freshness compared to the untreated ones after 3 days (Fig. 5 A, B, and C). The experiment was also conducted with grapes over a
period of 5 days (Fig. 6 A, B, and C) and with bananas over 3 days, yielding similar positive outcomes. These findings indicate that the
CFSs of the LAB possess the potential to serve as a natural preservative (Fig. 7 A, B, and C).

3.11. Preservation of sugarcane juice

3.11.1. Changes in pH levels with fermentation

The pH levels of the test samples were measured and are presented here in Fig. 8. The level of pH decreased with the passing of time
due to the increase of the Lactobacillus plantarum DMR14. Initially, the pH was 6.5 and, by the end, the final pH had reached 3.8 for the
control and 4.8 for the CFS-treated juice.

3.11.2. Cell viability of LAB cultures during fermentation

Fermentation of sugarcane juice with lactic acid bacteria may have beneficial health effects on the body due to metabolites pro­
duced by the bacteria. With the passing of time, the concentration of the lactic acid bacteria increased gradually, indicating that
fermented sugarcane juice could be a good career of LAB (Fig. 9A and Fig. S2). Furthermore, E. coli levels decreased, indicating that
Lactobacillus plantarum DMR14 has anti-bacterial efficacy (Fig. 9B and Fig. S3).

4. Discussion

Lactic acid bacteria (LAB) are widely recognized as safe microorganisms, with the United States Food and Drug Administration

Table 6
Antagonistic activity of the Lactobacillus plantarum DMR14 against six selected pathogenic bacterial strains.
Bacterial Name Zone of Inhibition (mm) Status

Shigella boydii 17 ± 1 Moderate/average inhibition (++)

Pseudomonas sp. 12 ± 1 Weak inhibition (+)
Escherichia coli 7.33 ± 0.57 No inhibitory activity (− )
Bacillus cereus 6 No inhibitory activity (− )
Aeromonas sp. 12.67 ± 1 Weak inhibition (+)
Staphylococcus aureus 13 ± 0.57 Weak inhibition (+)

Note: 12–15 mm, weak inhibition (+); 16–19 mm, moderate/average inhibition.

7
S. Islam et al. Heliyon 9 (2023) e17382

Table 7
The biofilm formation efficacy of the selected bacterial strains.
Name of the Bacteria Range Result

Shigella boydii OD > 4 × ODcutoff Strong

Pseudomonas sp. OD > 4 × ODcutoff Strong
Staphylococcus aureus OD > 4 × ODcutoff Strong
Escherichia coli OD > 4 × ODcutoff Strong
Bacillus cereus OD > 4 × ODcutoff Strong
Aeromonas sp. OD > 4 × ODcutoff Strong

Fig. 3. Biofilm inhibition percentage of the cell free supernatants (CFS) of the Lactobacillus plantarum DMR14 against the selected bacterial strains.
The values were reported in optical density (OD) as mean ± SD of three replicates.

Fig. 4. Extending shelf-life of apple with the CFS of Lactobacillus plantarum DMR14 (A) Fresh apple at 0 h, (B) after 7 days without treatment; (C)
after 7 days with treatment.

Fig. 5. Extending shelf-life of chopped apples with the cell free supernatants (CFS) from Lactobacillus plantarum DMR14. (A) Fresh chopped apple at
0 h, (B) and (C) after 3 days without treatment and with treatment, respectively.

8
S. Islam et al. Heliyon 9 (2023) e17382

Fig. 6. Extending shelf-life of grapes with the cell free supernatants (CFS) of Lactobacillus plantarum DMR14. (A) Fresh grapes at 0 h, (B), and (C)
after 5 days without treatment and with treatment, respectively.

Fig. 7. Extending shelf-life of the bananas with the cell free supernatants (CFS) of Lactobacillus plantarum DMR14. (A) Fresh banana at 0 h, (B), and
(C) after 3 days without treatment and with treatment, respectively.

Fig. 8. Change in pH levels with the fermentation of sugarcane juice with 100 μl of cell free supernatants (CFS) of the Lactobacillus plantarum
DMR14. Data were recorded 24 h of interval.

(FDA) classifying them as generally recognized as safe (GRAS) and the European Food Safety Authority (EFSA) considering them to be
under the qualified presumption of safety (QPS) (EFSA). Given their safety status, LAB has become an attractive option for food
biopreservation as an alternative method for preventing the growth of pathogenic microorganisms. Due to their competitive edge
against pathogenic bacteria, LAB is ideal candidates for developing bioprotective agents for fresh fruits, vegetables, and fruit juice. This
natural preservation approach not only increases the shelf life of these perishable food products but also helps to maintain their
nutritional content while eliminating the need for synthetic preservatives that may have adverse effects on human health and the
environment [7,46]. In an effort to identify new sources of probiotic LAB for the preservation of fruits and fruit juice, the recent present

9
S. Islam et al. Heliyon 9 (2023) e17382

Fig. 9. Viability of bacteria on CFS treated juice. (A) Lactic acid bacteria, (B) coliform bacteria. Data were recorded in various time durations (0 h,
24 h, 48 h, and 72 h).

study focused on isolating LAB from fermented oats. The study aimed to demonstrate the potential of cereal-based meals as a source of
probiotic LAB, particularly for individuals who are unable to consume dairy-based products due to lactose intolerance or allergies. One
of the key objectives of the study was to evaluate the probiotic potential of the isolated LAB strains and their ability to preserve fruits
and fruit juice naturally. The findings suggest that the LAB strains isolated from fermented oats have the potential to be used as
probiotics and natural preservatives, providing a promising alternative to conventional preservatives. By utilizing a non-dairy source
of probiotic LAB, this approach could provide an option for individuals with dietary restrictions while promoting sustainable and
healthier food preservation practices. In this study, Lactobacillus plantarum DMR14 was isolated from fermented oats using MRS se­
lective media and was identified through 16S rRNA gene sequencing (as shown in Fig. 1). To determine its potential as a probiotic
strain, the isolate was subjected to various criteria, including antagonistic efficacy, pH tolerance, bile salt tolerance, antibiotic sus­
ceptibility, among others. Among these criteria, pH tolerance is a crucial factor for determining the viability of probiotic strains.
Probiotic bacteria should have the ability to survive in the highly acidic environment of the stomach and tolerate low pH values as low
as pH 2.0 [47]. Thus, the pH tolerance of L. plantarum DMR14 was evaluated as part of the study’s objective to assess its probiotic
potential. Here, Lactobacillus plantarum DMR14 showed a wide range of pH tolerance (ranging from 2 to 7), as well as bile salt
resistance, indicating that it has wide range of pH tolerance. Furthermore, another most important characteristic of probiotics is
resistance to bile salts since these substances breakdown membrane lipids, which results in cell leakage and death. The acid and bile
salt tolerance for the LAB varied significantly due to the specific nature of the strain, and thus depended on species. In previous studies,
L. plantarum and E. lactis showed acceptable survival in a bile salt environment [43] even in a high concentration (0.3%). In this study,
by being tolerant to high bile salt concentration (0.3%) the isolated strain full filled other criteria of the probiotic strains.
Antibiotic resistance is one of the key parameters for considering any microbe to be safe [48]. However, the genetic mechanisms for
antibiotic susceptibility in LAB strains are limited. There are several genes in several species of the LAB that are responsible for
antibiotic resistance [49]. Lactobaccilus are usually sensitive to the cell wall-targeting penicillin and susceptible to antibiotics that
inhibit protein synthesis, such as chloramphenicol, lincomycin and tetracyclin. Here, Lactobacillus plantarum DMR14 was found to be
resistant against amoxicillin (AMX) and erythromycin (E) and showed intermediate resistance to penicillin (P) and ampicillin (AMP).
From this result, it is clear that antibiotic susceptibility and resistance of LAB is also varying with different species. Overall, the findings
suggest that L. plantarum DMR14 exhibits promising probiotic properties and has the potential to be used as a natural preservative for
fruits and fruit juice.
Hemolysis activity testing is one of the necessary probiotic assays. The gamma-hemolytic potentiality of Lactobacillus plantarum
DMR14 indicated that it is not toxic to the environment or to individuals. Additionally, the strain’s test results for lipolytic and
proteolytic activity were both favorable. Similar results were observed by Hassan Barzegar regarding Lactobacillus plantarum DMR14
[50]. Antagonistic activity of the bacteria is generally tested to use it as a natural killer of the pathogenic bacteria. Here, Lactobacillus
plantarum DMR14 showed strong inhibition activity against Shigella boydii and moderate antagonistic efficacy against Pseudomonas sp.,
Aeromonas sp., and Staphylococcus aureus. Antagonistic activity of the strain was also observed in previous studies, which is in support
of our outcomes [51,52]. Biofilm formation is one of the main causes of antibiotic resistance [53]. Lactobacillus plantarum DMR14
showed positive results for biofilm inhibition potential against all selected strains. Like in the antagonistic testing, the most prominent
result in this case was also observed against Shigella boydii. Hojjatolah Zamani [54] also reported Lactobacillus plantarum DMR14’s
biofilm inhibition efficacy against Staphylococcus aureus, and Escherichia coli, which is consistent with our findings. Natural pre­
servatives made from the healthy Lactobacillus plantarum DMR14 strain are particularly sought after because chemical preservatives
are harmful to our health. Bacteriocin is a product of LAB that primarily serves as a preservative. Additionally, the human body’s
proteolytic enzymes can degrade bacteriocin, making this bacterial treatment safe for our health [55].
After confirming bacterial probiotic potentiality, the main applied sector of the work was to use it as a natural preservative for the
fruit industries. So, in this study, the potential of Lactobacillus plantarum DMR14 to increase the shelf life of fruits was investigated by
immersing apples in its CFS. The LAB strain was found to effectively prevent the rotting of apples even after seven days of storage.
Sliced apples that were untreated turned yellow and dried out, while the coated ones maintained their original color and freshness. The
same positive results were observed with bananas and grapes. The findings of this study were consistent with the results of previous

10
S. Islam et al. Heliyon 9 (2023) e17382

research conducted by S. Agriopoulou et al. and Fang et al. who also used different strains of bacteria, including Salimicrobium, Bacillus,
Paenibacillus, Lactobacillus delbrueckii subsp. bulgaricus, and Leuconostoc lactis, to preserve fruits such as apples, bananas, strawberry
and grapes [56–58]. Additionally, it has been shown that Lactobacillus equigeneosi and Bacillus Smithii are effective at increasing the
shelf life of pomegranates [45].
In a trial investigation, sugarcane juice, one of the popular street juices in the country, was used as a medium to assess the suitability
of LAB as a natural preservative. The study aimed to determine whether the CFS of Lactobacillus plantarum DMR14 could be used as a
powerful preservative for the juice. The results showed that the pH of the treated juice declined gradually compared to the control,
while the concentration of LAB steadily rose, indicating that LAB could be used as a natural preservative for sugarcane juice. Similar
results for orange juice and tomato juice were found in another study [59]. Qiao, Huahua, demonstrated that the sugarcane juice
treated with Lactobacillus HNK10 and cultured at 37 ◦ C for 48 h contained a significant amount of LAB, as well as high-quality and
potent antibacterial activity, which supports our findings [60]. In our research, we also observed that CFU coliform bacteria in the juice
treated with CFS dropped steadily. This result indicates that harmful bacteria declined as LAB increased. Thus, the CFS of Lactobacillus
plantarum DMR14 may be employed in juice industries. However, no studies were found in which the researchers attempted to
measure the concentration of pathogenic bacteria in LAB-fortified juice.

5. Conclusion

In these study, the probiotic lactic acid bacteria (LAB) from fermented oats was isolated to address the demand for natural pre­
servatives. Here, Lactobacillus plantarum DMR14 strain showed strong antimicrobial and antibiofilm activity against Shigella boydii.
Moreover, the strain’s cell-free supernatant (CFS) extended the shelf life of the treated fruits and sugarcane juice. Here, the number of
coliform bacteria decreased, while lactic acid bacteria increased in the CFS-treated sugarcane juice. The findings suggest Lactobacillus
plantarum DMR14 as a potential fruit preservative in the food industry. This research contributes to the search for safer alternatives to
chemical preservatives. Further exploration of probiotic-based preservation methods can benefit human health and the environment.

Author contribution statement

Shirmin Islam, Md. Abu Saleh, Shahriar Zaman: Conceived and designed the experiments; Performed the experiments; Wrote the
paper. Suvro Biswas, Tabassum Jabin, Md. Moniruzzaman, Jui Biswas: Performed the experiments; Analyzed and interpreted the data;
Wrote the paper. Md. Salah Uddin, Md. Akhtar-E-Ekram, Abdallah M. Elgorban, Gajanan Ghodake, Asad Syed: Contributed reagents,
materials, analysis tools or data; Wrote the paper.

Data availability statement

Data will be made available on request.

Funding

This work was supported by the Grants for Advanced Research in Education (Grant Number: LS20222099), Ministry of Education,
Government of the People’s Republic of Bangladesh. It was also supported by the Researchers Supporting Project Number (RSP-2021/
367), King Saud University, Riyadh, Saudi Arabia.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to
influence the work reported in this paper.

Acknowledgments

The authors extend their appreciation to the Researchers Supporting Project Number (RSP-2021/367), King Saud University,
Riyadh, Saudi Arabia.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.heliyon.2023.e17382.

References

[1] P.P. Joy, Fruits , Benefits , Processing , Preservation and Pineapple Recipes, 2016.

11
S. Islam et al. Heliyon 9 (2023) e17382

[2] H. Zhang, G.K. Mahunu, R. Castoria, M.T. Apaliya, Q. Yang, Augmentation of biocontrol agents with physical methods against postharvest diseases of fruits and
vegetables, Trends Food Sci. Technol. 69 (2017) 36–45, https://doi.org/10.1016/j.tifs.2017.08.020.
[3] F. Chemat, Zill-E-Huma, M.K. Khan, Applications of ultrasound in food technology: processing, preservation and extraction, Ultrason. Sonochem. 18 (2011)
813–835, https://doi.org/10.1016/j.ultsonch.2010.11.023.
[4] M. Bilal, S. Mehmood, H.M.N. Iqbal, The beast of beauty: environmental and health concerns of toxic components in cosmetics, Cosmetics 7 (2020), https://doi.
org/10.3390/cosmetics7010013.
[5] A. Meireles, E. Giaouris, M. Simões, Alternative disinfection methods to chlorine for use in the fresh-cut industry, Food Res. Int. 82 (2016) 71–85, https://doi.
org/10.1016/j.foodres.2016.01.021.
[6] S.P. Anand, N. Sati, Artificial preservatives and their harmful effects: looking toward nature for safer alternatives, International Journal of Pharmaceutical
Sciences and Research IJPSR 4 (2013) 2496–2501, https://doi.org/10.13040/IJPSR.0975-8232.4(7).24960-01.
[7] S. Agriopoulou, E. Stamatelopoulou, M. Sachadyn-Król, T. Varzakas, Lactic acid bacteria as antibacterial agents to extend the shelf life of fresh and minimally
processed fruits and vegetables: quality and safety aspects, Microorganisms 8 (2020) 1–23, https://doi.org/10.3390/microorganisms8060952.
[8] A. Akbar, I. Ali, A.K. Anal, Industrial perspectives of lactic acid bacteria for biopreservation and food safety, Journal of Animal and Plant Sciences 26 (2016)
938–948.
[9] P.M. De Oliveira, B.R. Leite Júnior, M.L. Martins, E.M.F. Martins, A.M. Ramos, Minimally processed yellow melon enriched with probiotic bacteria, Semina
Ciências Agrárias 35 (2014) 2415–2426, https://doi.org/10.5433/1679-0359.2014v35n5p2415.
[10] M.E. Kasimin, S. Shamsuddin, A.M. Molujin, M.K. Sabullah, J.A. Gansau, R. Jawan, Enterocin: promising biopreservative produced by Enterococcus sp,
Microorganisms 10 (2022), https://doi.org/10.3390/microorganisms10040684.
[11] B. Ramos, F.A. Miller, T.R.S. Brandão, P. Teixeira, C.L.M. Silva, Fresh fruits and vegetables - an overview on applied methodologies to improve its quality and
safety, Innovative Food Sci. Emerging Technol. 20 (2013) 1–15, https://doi.org/10.1016/j.ifset.2013.07.002.
[12] A. Terpou, A. Papadaki, I.K. Lappa, V. Kachrimanidou, L.A. Bosnea, N. Kopsahelis, Probiotics in food systems: significance and emerging strategies towards
improved viability and delivery of enhanced beneficial value, Nutrients 11 (2019), https://doi.org/10.3390/nu11071591.
[13] M.P. Mokoena, Lactic acid bacteria and their bacteriocins: classification, biosynthesis and applications against uropathogens: a mini-review, Molecules 22
(2017), https://doi.org/10.3390/molecules22081255.
[14] N. Vieco-Saiz, Y. Belguesmia, R. Raspoet, E. Auclair, F. Gancel, I. Kempf, D. Drider, Benefits and inputs from lactic acid bacteria and their bacteriocins as
alternatives to antibiotic growth promoters during food-animal production, Front. Microbiol. 10 (2019), https://doi.org/10.3389/fmicb.2019.00057.
[15] S.J. Lee, H.S. Jeon, J.Y. Yoo, J.H. Kim, Some important metabolites produced by lactic acid bacteria originated from kimchi, Foods 10 (2021), https://doi.org/
10.3390/foods10092148.
[16] C. Hill, F. Guarner, G. Reid, G.G.-N.R, The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate
use of the term probiotic, Nature.Com (2014) 1–5, undefined 2014, https://www.nature.com/articles/nrgastro.2014.66%22.
[17] T. Chen, L. Wang, Q. Li, Y. Long, Y. Lin, J. Yin, Y. Zeng, L. Huang, T. Yao, M.N. Abbasi, H. Yang, Q. Wang, C. Tang, T.A. Khan, Q. Liu, J. Yin, Q. Tu, Y. Yin,
Functional probiotics of lactic acid bacteria from Hu sheep milk, BMC Microbiol. 20 (2020), https://doi.org/10.1186/s12866-020-01920-6.
[18] J.M. Hess, S.S. Jonnalagadda, J.L. Slavin, Dairy foods: current evidence of their effects on bone, cardiometabolic, cognitive, and digestive health, Compr. Rev.
Food Sci. Food Saf. 15 (2016) 251–268, https://doi.org/10.1111/1541-4337.12183.
[19] P. Tsafrakidou, A.M. Michaelidou, C.G. Biliaderis, Fermented cereal-based products: nutritional aspects, possible impact on gut microbiota and health
implications, Foods 9 (2020), https://doi.org/10.3390/FOODS9060734.
[20] S.B. Lim, J.M.R. Tingirikari, J.S. Seo, L. Li, S. Shim, J.H. Seo, N.S. Han, Isolation of lactic acid bacteria starters from Jeung-pyun for sourdough fermentation,
Food Sci. Biotechnol. 27 (2018) 73–78, https://doi.org/10.1007/s10068-017-0274-0.
[21] I.N. Sujaya, S. Amachi, A. Yokota, K. Asano, F. Tomita, Identification and characterization of lactic acid bacteria in ragi tape, World J. Microbiol. Biotechnol. 17
(2001) 349–357, https://doi.org/10.1023/A:1016642315022.
[22] S.M. Thwe, T. Kobayashi, T. Luan, T. Shirai, M. Onodera, N. Hamada-Sato, C. Imada, Isolation, characterization, and utilization of γ-aminobutyric acid (GABA)-
producing lactic acid bacteria from Myanmar fishery products fermented with boiled rice, Fisheries Science 77 (2011) 279–288, https://doi.org/10.1007/
s12562-011-0328-9.
[23] K. Väkeväinen, A. Valderrama, J. Espinosa, D. Centurión, J. Rizo, D. Reyes-Duarte, G. Díaz-Ruiz, A. von Wright, P. Elizaquível, K. Esquivel, A.I. Simontaival,
R. Aznar, C. Wacher, C. Plumed-Ferrer, Characterization of lactic acid bacteria recovered from atole agrio, a traditional Mexican fermented beverage, LWT -
Food Sci. Technol. (Lebensmittel-Wissenschaft -Technol.) 88 (2018) 109–118, https://doi.org/10.1016/j.lwt.2017.10.004.
[24] T.T.T. Nguyen, G. Loiseau, C. Icard-Vernière, I. Rochette, S. Trèche, J.P. Guyot, Effect of fermentation by amylolytic lactic acid bacteria, in process
combinations, on characteristics of rice/soybean slurries: a new method for preparing high energy density complementary foods for young children, Food Chem.
100 (2007) 623–631, https://doi.org/10.1016/j.foodchem.2005.09.080.
[25] A. Blandino, M.E. Al-Aseeri, S.S. Pandiella, D. Cantero, C. Webb, Cereal-based fermented foods and beverages, Food Res. Int. 36 (2003) 527–543, https://doi.
org/10.1016/S0963-9969(03)00009-7.
[26] C.M.B.K. Muyanja, J.A. Narvhus, J. Treimo, T. Langsrud, Isolation, characterisation and identification of lactic acid bacteria from bushera: a Ugandan
traditional fermented beverage, Int. J. Food Microbiol. 80 (2003) 201–210, https://doi.org/10.1016/S0168-1605(02)00148-4.
[27] F. Manini, M.C. Casiraghi, K. Poutanen, M. Brasca, D. Erba, C. Plumed-Ferrer, Characterization of lactic acid bacteria isolated from wheat bran sourdough, LWT -
Food Sci. Technol. (Lebensmittel-Wissenschaft -Technol.) 66 (2016) 275–283, https://doi.org/10.1016/j.lwt.2015.10.045.
[28] A. Corsetti, P. Lavermicocca, M. Morea, F. Baruzzi, N. Tosti, M. Gobbetti, Phenotypic and molecular identification and clustering of lactic acid bacteria and
yeasts from wheat (species Triticum durum and Triticum aestivum) sourdoughs of Southern Italy, Int. J. Food Microbiol. 64 (2001) 95–104, https://doi.org/
10.1016/S0168-1605(00)00447-5.
[29] L. De Vuyst, S. Van Kerrebroeck, H. Harth, G. Huys, H.M. Daniel, S. Weckx, Microbial ecology of sourdough fermentations: diverse or uniform? Food Microbiol.
37 (2014) 11–29, https://doi.org/10.1016/j.fm.2013.06.002.
[30] I. Ganchev, Z. Koleva, Y. Kizheva, P. Moncheva, P. Hristova, Lactic acid bacteria from spontaneosly fermented rye sourdough, Bulgarian Journal of Agricultural
Science 20 (2014) 69–73.
[31] D. Savić, T. Savić, M. Škrinjar, N. Joković, Profile of lactic acid bacteria in rye flour and sourdough, J. Cult. Collect. 5 (2007) 38–45. http://www.bioline.org.br/
request?cc06006%5Cnfile:///Users/richiepreiss/Library/Application%20Support/Papers2/Articles/2007/Savić/2007%20Savić.pdf%5Cnpapers2://
publication/uuid/7B894ED1-2047-4908-9F93-87B94EE12C65.
[32] T. Onda, F. Yanagida, T. Uchimura, M. Tsuji, S. Ogino, T. Shinohara, K. Yokotsuka, Analysis of lactic acid bacterial flora during Miso fermentation, Food Sci.
Technol. Res. 9 (2003) 17–24, https://doi.org/10.3136/fstr.9.17.
[33] O. Mårtensson, R. Öste, O. Holst, Texture promoting capacity and EPS formation by lactic acid bacteria in three different oat-based non-dairy media, Eur. Food
Res. Technol. 214 (2002) 232–236, https://doi.org/10.1007/s00217-001-0440-7.
[34] A. Angelov, V. Gotcheva, R. Kuncheva, T. Hristozova, Development of a new oat-based probiotic drink, Int. J. Food Microbiol. 112 (2006) 75–80, https://doi.
org/10.1016/j.ijfoodmicro.2006.05.015.
[35] A.Y. Bustos, C.L. Gerez, L.G. Mohtar Mohtar, V.I. Paz Zanini, M.A. Nazareno, M.P. Taranto, L.B. Iturriaga, Lactic acid fermentation improved textural behaviour,
phenolic compounds and antioxidant activity of Chia (Salvia hispanica L.) dough, Food Technol. Biotechnol. 55 (2017) 381–389, https://doi.org/10.17113/ft
b.55.03.17.5133.
[36] D. Arpita, R. Utpal, C. Runu, Cereal based functional food of Indian subcontinent: a review, J. Food Sci. Technol. 49 (2012) 665–672. http://ovidsp.ovid.com/
ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=cagh&AN=201233391. 00%5Cnhttp://lshtmsfx. .hosted.exlibrisgroup.com/lshtm?sid=OVID:
caghdb&id=pmid:&id=doi:&issn=0022-1155&isbn=&volume=49&issue=6&spage=665&pages=665-672&date=2012&title=Jour.
[37] M. Lee, M. Regu, S. Seleshe, Uniqueness of Ethiopian traditional alcoholic beverage of plant origin, tella, Journal of Ethnic Foods 2 (2015) 110–114, https://doi.
org/10.1016/j.jef.2015.08.002.

12
S. Islam et al. Heliyon 9 (2023) e17382

[38] M. Naeem, I. Ahmed, S. Ahmed, Z. Ahmed, M.N. Riaz, S. Ghazanfar, Screening of cattle gut associated Bacillus strains for their potential use as animal probiotic,
Indian J. Anim. Res. (2018), https://doi.org/10.18805/ijar.b-948.
[39] G. Mulaw, T. Sisay Tessema, D. Muleta, A. Tesfaye, In vitro evaluation of probiotic properties of lactic acid bacteria isolated from some traditionally fermented
ethiopian food products, International Journal of Microbiology (2019) 2019, https://doi.org/10.1155/2019/7179514.
[40] F. Gashe, E. Mulisa, M. Mekonnen, G. Zeleke, Antimicrobial resistance profile of different clinical isolates against third-generation cephalosporins, Journal of
Pharmaceutics (2018) 1–7, https://doi.org/10.1155/2018/5070742, 2018.
[41] B.R. Shah, B. Li, H. Al Sabbah, W. Xu, J. Mráz, Effects of prebiotic dietary fibers and probiotics on human health: with special focus on recent advancement in
their encapsulated formulations, Trends Food Sci. Technol. 102 (2020) 178–192, https://doi.org/10.1016/j.tifs.2020.06.010.
[42] S. Mahmud, G.K. Paul, M. Afroze, S. Islam, S.B.R. Gupt, M.H. Razu, S. Biswas, S. Zaman, M.S. Uddin, M. Khan, N.A. Cacciola, T. Bin Emran, M.A. Saleh,
R. Capasso, J. Simal-Gandara, Efficacy of phytochemicals derived from avicennia officinalis for the management of covid-19: a combined in silico and
biochemical study, Molecules 26 (2021), https://doi.org/10.3390/molecules26082210.
[43] A. Sharma, M. Lavania, R. Singh, B. Lal, Identification and probiotic potential of lactic acid bacteria from camel milk, Saudi J. Biol. Sci. 28 (2021) 1622–1632,
https://doi.org/10.1016/j.sjbs.2020.11.062.
[44] O. GA, Microtiter dish biofilm formation assay, JoVE (2011) 2437.
[45] V. Dhundale, V. Hemke, D. Desai, P. Dhundale, Evaluation and exploration of lactic acid bacteria for preservation and extending the shelf life of fruit, Int. J.
Fruit Sci. 18 (2018) 355–368, https://doi.org/10.1080/15538362.2018.1435331.
[46] J.R. Linares-Morales, N. Gutiérrez-Méndez, B.E. Rivera-Chavira, S.B. Pérez-Vega, G.V. Nevárez-Moorillón, Biocontrol processes in fruits and fresh produce, the
use of lactic acid bacteria as a sustainable option, Front. Sustain. Food Syst. 2 (2018), https://doi.org/10.3389/fsufs.2018.00050.
[47] C. Prabhurajeshwar, R.K. Chandrakanth, Probiotic potential of Lactobacilli with antagonistic activity against pathogenic strains: an in vitro validation for the
production of inhibitory substances, Biomed. J. 40 (2017) 270–283, https://doi.org/10.1016/j.bj.2017.06.008.
[48] C.X. Hiller, U. Hübner, S. Fajnorova, T. Schwartz, J.E. Drewes, Antibiotic microbial resistance (AMR) removal efficiencies by conventional and advanced
wastewater treatment processes: a review, Sci. Total Environ. 685 (2019) 596–608, https://doi.org/10.1016/j.scitotenv.2019.05.315.
[49] L. Solieri, A. Bianchi, G. Mottolese, F. Lemmetti, P. Giudici, Tailoring the probiotic potential of non-starter Lactobacillus strains from ripened Parmigiano
Reggiano cheese by invitro screening and principal component analysis, Food Microbiol. 38 (2014) 240–249, https://doi.org/10.1016/j.fm.2013.10.003.
[50] H. Barzegar, B. Alizadeh Behbahani, F. Falah, Safety, probiotic properties, antimicrobial activity, and technological performance of Lactobacillus strains isolated
from Iranian raw milk cheeses, Food Sci. Nutr. 9 (2021) 4094–4107, https://doi.org/10.1002/fsn3.2365.
[51] S. El oirdi, T. Lakhlifi, A.A. Bahar, M. Yatim, Z. Rachid, A. Belhaj, Isolation and identification of Lactobacillus plantarum 4F, a strain with high antifungal
activity, fungicidal effect, and biopreservation properties of food, J. Food Process. Preserv. 45 (2021), https://doi.org/10.1111/jfpp.15517.
[52] N. Karimi, V. Jabbari, A. Nazemi, K. Ganbarov, N. Karimi, A. Tanomand, S. Karimi, A. Abbasi, B. Yousefi, E. Khodadadi, H.S. Kafil, Thymol, cardamom and
Lactobacillus plantarum nanoparticles as a functional candy with high protection against Streptococcus mutans and tooth decay, Microb. Pathog. 148 (2020),
https://doi.org/10.1016/j.micpath.2020.104481.
[53] G. Gebreyohannes, A. Nyerere, C. Bii, D.B. Sbhatu, Challenges of intervention, treatment, and antibiotic resistance of biofilm-forming microorganisms, Heliyon
5 (2019), https://doi.org/10.1016/j.heliyon.2019.e02192.
[54] H. Zamani, S. Rahbar, S.R. Garakoui, A. Afsah Sahebi, H. Jafari, Antibiofilm potential of Lactobacillus plantarum spp. cell free supernatant (CFS) against
multidrug resistant bacterial pathogens, Pharmaceutical and Biomedical Research 3 (2017) 39–44, https://doi.org/10.29252/pbr.3.2.39.
[55] H.P. Rupasinghe, L.J. Yu, Emerging Preservation Methods for Fruit Juices and Beverages, Food Additive, 2012, pp. 65–82.
[56] X. Fang, Y. Li, W. Guo, W. Ke, S. Bi, X. Guo, Y. Zhang, Lactobacillus delbrueckii subsp. bulgaricus F17 and Leuconostoc lactis H52 supernatants delay the decay
of strawberry fruits: a microbiome perspective, Food Funct. 10 (2019) 7767–7781, https://doi.org/10.1039/c9fo02079a.
[57] S. Agriopoulou, E. Stamatelopoulou, M. Sachadyn-Król, T. Varzakas, Lactic acid bacteria as antibacterial agents to extend the shelf life of fresh and minimally
processed fruits and vegetables: quality and safety aspects, Microorganisms 8 (2020) 1–23, https://doi.org/10.3390/microorganisms8060952.
[58] X. Fang, Q. Duan, Z. Wang, F. Li, J. Du, W. Ke, D. Liu, R.C. Beier, X. Guo, Y. Zhang, Products of lactobacillus delbrueckii subsp. Bulgaricus strain f17 and
leuconostoc lactis strain h52 are biopreservatives for improving postharvest quality of ‘red globe’ grapes, Microorganisms 8 (2020), https://doi.org/10.3390/
microorganisms8050656.
[59] R. Nagpal, A. Kumar, M. Kumar, Fortification and fermentation of fruit juices with probiotic lactobacilli, Ann. Microbiol. 62 (2012) 1573–1578, https://doi.org/
10.1007/s13213-011-0412-5.
[60] H. Qiao, L. Chen, J. Yang, W. Zhi, R. Chen, T. Lu, H. Tan, Z. Sheng, Effect of lactic acid bacteria on bacterial community structure and characteristics of
sugarcane juice, Foods 11 (2022), https://doi.org/10.3390/foods11193134.

13

View publication stats