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Investigation of Probiotic Features of Bacteria Isolated from Some
Food Products
Murat DOGAN 1 Haydar OZPINAR 1
1 Istanbul Aydin Univeristy, Faculty of Engineering, Department of Food Engineering, TR-34295 Istanbul - TURKEY
Article Code: KVFD-2016-17273 Received: 19.12.2016 Accepted: 10.03.2017 Published Online: 11.03.2017
Citation of This Article
Dogan M, Ozpinar H: Investigation of probiotic features of bacteria isolated from some food products. Kafkas Univ Vet Fak Derg, 23 (4): 555-562,
2017. DOI: 10.9775/kvfd.2016.17273
Abstract
Probiotics are used as natural supplements for good health and treatment of various diseases. Probiotics aect health in a positive way
due to their activities in the gastrointestinal tract. There is a growing interest in using probiotic bacteria for their protective eects against
diseases and an emerging trend towards consuming healthy foods. The aim of the present study was to reveal species with alternative
probiotic properties from some food products, which are already known to have probiotic properties and whose natural properties are
preserved. Probiotic characteristics of isolated bacteria strains from 130 food samples which include 10 boza, 40 cheese, 20 kefir and 60 raw
milk samples were microbiologically analyzed in the present study.A total 144 strains including 127 Enterococcus faecium, 7 Lactobacillus
plantarum, 5 Lactobacillus para-plantarum and 5 Lactobacillus brevis were typed with characterizing by mass spectroscopy (MALDI-TOF MS)
to have probiotic eects.Then, all the tests required to comply with the probiotic properties of these bacteria were applied sequentially. Of
the 144 bacterial strains identified, only 35 were resistant to gastric pH. In the next step, only 8 isolates from 35 isolates were able to survive
under bile salt conditions. It has been determined that only 6 of bile salt-resistant isolates have the hydrophobicity ability. The remaining
6 isolates were examined for antimicrobial resistance and the presence of extended-spectrum beta-lactamases (ESBL) resistance and ESBL
were not detected. At the end of analysis, only 6 (4.1%) bacteria of 144 isolates were found to have probiotic properties. Three of them were
Lactobacillus brevis isolated from boza and 3 of them were Lactobacillus plantarum species isolated from kefir. However, no probiotics could
be isolated from other food samples such as milk and cheese. Therefore, the present study demonstrated that probiotic bacteria could be
produced as an alternative to industrial probiotics through non-transgenic microorganisms isolated from natural food products such as
kefir and boza.
Keywords: Probiotic Bacteria, Boza, Milk, Cheese, Kefir, Probiotic properties
Bazı Gıda Ürünlerinden İzole Edilen Bakterilerin Probiyotik Özelliklerinin
Araştırılması
Özet
Probiyotikler, çesitli hastalıkların tedavisi ve sağlık için doğal takviyeler olarak kullanılırlar. Probiyotikler gastrointestinal sistemde yaptıkları
faaliyetler sonucunda sağlığı olumlu yönde etkilemektedirler. Günümüzde hastalıklara karşı koruyucu etkileri ve sağlıklı gıdaların tüketimine
yönelik yoğun ilginin ortaya çıkışına bağlı olarak probiyotik bakterilere ilgi artmıştır. Bu araştırmada probiyotik özellikleri olduğu bilinen ve
doğal özellikleri korunmuş bazı gıdalardan probiyotik özelliklere sahip türlerin ortaya çıkartılması amaçlanmıştır. Araştırmamızda 10 boza, 40
peynir, 20 kefir ve 60 çiğ süt olmak üzere toplam 130 gıda örneği mikrobiyolojik bakımdan incelenmiştir. Sonuçta; 127 Enterococcus faecium,
7 Lactobacillus plantarum, 5 Lactobacillus para-plantarum ve 5 Lactobacillus brevis olmak üzere toplam 144 probiyotik etkisi gösterebilecek
bakteri kütle spektofotometre (MALDI-TOF MS) ile karakterize edilerek tiplendirilmiştir. Daha sonra bu bakterilerin probiyotik özelliklere
uygunluğu konusunda gerekli olan tüm testler sırasıyla uygulanmıştır. Karakterize edilen 144 izolattan sadece 35 ‘inin mide pH’sına dayanıklı
olduğu saptanmıştır. Bir sonraki basamakta ise yine 35 izolatdan sadece 8’i safra tuzu koşullarında canlılıklarını devam ettirebilmiştir. Safra
tuzuna dayanıklı izolatlardan sadece 6’sının hidrofobisite yeteneğine sahip olduğu belirlenmiştir. Kalan 6 izolatın antimikrobiyel direnç
durumları incelenmiş dirençliliğe ve ESBL varlığına rastlanmamıştır. İncelemeler sonunda 144 izolattan sadece 6 (4.1%) sının probiyotik
özelliklere sahip olduğu görülmüştür. Bunlardan; 3’ü Lactobacillus brevis bozadan ve 3’ü Lactobacillus plantarum kefirden izole edilmişlerdir.
Kefir ve bozadan probiyotik özellik gösteren bakteri izole edilirken diğer gıda örnekleri olan süt ve peynirden probiyotik özellik gösteren
bakteri izole edilememiştir. Sonuç olarak, kefir ve boza gibi doğal gıda ürünlerinden izole edilen transgenetik olmayan mikroorganizmalar
içinden probiyotik bakterilerin endüstriyel probiyotiklere alternatif olarak üretilebileceği bu araştırmada saptanmıştır.
Anahtar sözcükler: Probiyotik bakteri, Boza, Süt, Peynir, Kefir, Probiyotik özellik
İleşim (Correspondence)
+90 532 6226786
muratdogan72@gmail.com
KafKas Universitesi veteriner faKUltesi Dergisi
JoUrnal Home-Page: http://vetdergi.kafkas.edu.tr
online sUbmission: http://vetdergikafkas.org
Research Article
Kafkas Univ Vet Fak Derg
23 (4): 555-562, 2017
DOI: 10.9775/kvfd.2016.17273
556
Investigation of Probiotic Features of ...
INTRODUCTION
Probiotics are microorganisms that improve the
microbial balance of human and animal intestines and thus
are beneficial for the digestive system. These may be used
as a natural supplement for both the healthy development
of the body and the treatment and prevention of diseases.
The probiotic bacteria colonize on the surface of the
intestine by competing against pathogen microorganisms
for nutrients in the gastrointestinal system, thereby
positively affect the health [1,2]. A study has reported
that probiotics prevent Escherichia coli associated with
diarrhea and death in newborns [3]. It has been reported
that probiotics produce volatile fatty acids as a result of
fermentation by digesting dietary fibers: oligosaccharide
structures that remain undigested in the colon. In
addition, the formation of butyric acid inhibits colon
cancer [4]. Probiotics have been found to contribute to
lactose digestion due to the production of lactase enzyme;
stimulation of immune system with the enhancement of
IgA production; and to reduce allergens by inhibiting
the passage of antigen-presenting substances into the
circulatory system. Similarly, several reports highlight the
importance of preventive eects of probiotics on heart
diseases, hypertension, and urogenital diseases due to
anti-oxidant eects, the act of cell wall components like
angiotensin 1 enzyme inhibitors, and colonization on
urinary and vaginal surfaces [5]. Additionally, it has been
reported that Helicobacter pylori exert gastritis and ulcer-
inhibiting effects by the production of inhibitors and
preventive eect on hepatic encephalopathy formation
by inhibiting the urease-producing intestinal ora and
reducing the serum ammonia levels [6,7]. Various studies
report the use of Lactobacillus rhamnosus, a combination
of L. rhamnosus and Lactobacillus acidophilus for the
treatment of children with diarrhea, the prevention of
intestinal diseases, colon cancer and for the treatment of
heart diseases [8-10].
Therefore, it is essential to develop new probiotic
strains with dierent eects against dierent diseases and
to use them in preventive medicine.
There has been an increasing interest in the use
of probiotics due to their protective effects against
diseases. An expenditure of $28 million was reported on
research related to probiotic market and consumption
in USA in 2011 [11]. The increasing interest in probiotics
has accelerated the studies on the development of new
probiotic products.
The therapeutic results of probiotics have been found
to treat diseases such as colon cancer, ulcers and gastritis,
and allergies, diabetes. In one study, it was reported that
a commercial culture mixture obtained from Lactobacillus
species had the antiproliferative eect by inhibiting tumor
cells causing colon cancer [12]. A study has reported the
beneficial effects of Lactobacillus species, especially L.
rhamnosus and L. acidophilus against the infections caused
by Helicobacter pylori [13]. In an in vivo study with probiotic
L. brevis, an anti-allergic eect on anaphylaxis reduction
was observed [14]. Another study provided the evidence
that hemoglobin A1C and fasting blood glucose decreased
in diabetic patients after treatment with probiotic
supplements [15].
Probiotics obtained from nutrients should be able to
resist stomach acidity and bile salts and reach the intestinal
system alive to exert their beneficial eects. Further, they
should be able to colonize and survive on the epithelial
cell surfaces of intestinal mucosa [16].
The current study aimed to discover the alternative
species of bacteria with probiotic properties along with
preserved natural characteristics. Bacteria isolated from
boza, cheese, kefir, and raw milk samples were characterized
by Matrix Assisted Laser Desorption Ionization-Time of
Flight Mass Spectrometry (MALDI-TOF MS) (VITEK® MS) to
examine the probiotic properties of the single species. The
relationship between these parameters was established
according to Pearson’s nonparametric statistical correlation.
MATERIAL and METHODS
Materials
A total of 130 food samples consisting of 10 boza, 40
cheeses, 60 raw milk, and 20 kefir were obtained from
Marmara, Central and Eastern Anatolia regions of Turkey
between 2014 and 2016. The food samples were listed
in Table 1.
Methods
Isolation of Bacteria
de Man, Rogosa and Sharpe (MRS) agar, MRS broth,
M17 agar and M17 broth media were prepared and used
to isolate and identify the pure cultures of probiotics [17].
Identification with MALDI-TOF MS
The microorganisms were identified by using a system
formed by comparison with a reference spectrum obtained
from colonies formed on M17 and MRS agar. Matrix
Assisted Laser Desorption Ionization-Time of Flight Mass
Spectrometry (MALDI-TOF MS) (VITEK® MS) (bioMerieux,
France) was utilized to identify the protein profiles of cell
structures of the microorganisms [18].
Measurement of Acid Tolerance
In order to determine acid tolerance, the pH value
of MRS and M17 broths was reduced to 2.5 by using
hydrochloric acid (Sigma Aldrich, USA) for creating similar
environment to stomach acidity conditions. The viability
557
DOGAN, OZPINAR
of cultures was then monitored at pH 2.5. Colony growth
on solid media and broth turbidity were evaluated as
presence of the development [19].
Determination of Bile Salt Tolerance of Isolates
For the bile salt tolerance test, 0.3% (w/v) Oxgall
(Bile bovine, Sigma-Aldrich, USA) showing the antimicrobial
effect and containing conjugated and deconjugated
bile components was inoculated (1%) to 7 mL of MRS
and M17 broths. The viability was analyzed by colony
counting and broth turbidity after 48-72 h incubation
at 37°C [20].
Determination of Hydrophobicity of Isolates
Active cultures in MRS and M17 broths were centrifuged
for 15 min at 10.000 rpm. The resulting pellet was washed
twice with phosphate buer, dissolved in 0.1 M KNO3 (pH
6.2) buer, added to 96-well plates, and OD was set to 600
nm using a spectrophotometer (A0). The cell suspension
(1 mL) was mixed with 0.3 mL of xylene and incubated at
room temperature for 4 h. Subsequently, the OD of the
aqueous phase was measured again at 600 nm (A1) and
the microbial adhesion of isolates to hydrocarbons was
determined using the formula [(A0-A1)/A0] x100 [21].
Antibiotic Susceptibility Test
Disc diffusion method was utilized for antibiotic
susceptibility analysis. Antibiogram verification and
determination of MIC (Minimal Inhibitory Concentration)
were performed by using Micronaut-S beta-lactamase
VII Plate (Merlin Diagnostika, Germany) according to the
phenotypic determination to identify the presence of
ESBL with MIC parameters [22].
Statistical Analyses
Statistical analyses were performed by SPSS Inc.
Software (22.0 Version, SPSS Inc., Chicago, IL). In the
statistical analysis, Pearson’s correlation was used to
examine whether all the data correlated with each other.
RESULTS
Isolation of bacteria from a total of 130 food samples,
including boza, cheese, kefir, and raw milk with MALDI-TOF
MS resulted in as of L. brevis, L. plantarum, L. para plantarum,
and E. faecium species. Among the 144 identified probiotic
isolates (five L. brevis, seven L. plantarum, five L. para
plantarum, and 127 E. faecium), 35 (five L. brevis, five L.
plantarum, three L. para plantarum, and 22 E. faecium)
passed the gastric pH resistance test. Out of the 35
isolates, eight isolates (four L. brevis, three L. plantarum,
and one E. faecium) could resist stomach pH and maintain
the viability in bile salt conditions in the gastrointestinal
tract, whereas only six isolates (three L. brevis and three
L. plantarum) displayed hydrophobicity. The remaining
six isolates (three L. brevis and three L. plantarum) were
analyzed for antimicrobial resistance according to the
instructions of the Institute for Clinical and Laboratory
Standards, and resistance or ESBLs were not detected.
The study concluded that only six (4.1%) of a total of 144
probiotic bacteria exhibited probiotic properties. L. brevis
and L. plantarum, the bacteria isolated from kefir and boza,
were able to companced the criteria of probiotics [23,24].
The relationship between test parameters was
determined according to Pearson’s nonparametric
statistical correlation, which revealed that there was a
significant correlation between acid and bile salt tolerance
of the isolates (P<0.05). Results are listed in Table 2, Table 3,
Table 4, Fig. 1, Fig. 2 and Fig. 3.
DISCUSSION
Only six (4.1%) isolates with probiotic properties were
detected among 144 isolates obtained from food sources.
Three of them were L. brevis strains isolated from boza,
and the others were L. plantarum strains from kefir. In the
previous studies, Lactobacillus spp. isolates with similar
probiotic properties to our study were isolated from kefir
and boza samples [23,24].
Similarly, Yadav (2016) isolated 54 strains belonging to
L. plantarum which were obtained from a local fermented
food from grain, stomach acidity and bile salts were
checked. It was determined that all isolates showed poor
resistance. Only 24 isolates (44%) were able to show good
resistance. Six (11%) species that could remain viable were
analyzed for probiotic properties, and L. plantarum RYPR1
(1.9%) exhibited satisfying results [25].
In the present study, the bacteria obtained from raw
Table 1. Distribution of food samples
Region Type of Food
BozaaCheesebRaw MilkcKefird
Marmara 10 10 * 10 * 5 *
Central Anatolia - 5 * + 5 ** 17 * + 13 ** 10 *
Eastern Anatolia - 9 * + 11 ** 10 * + 10 ** 5 *
a,b,c,d natural, non industrial type and non using starter culture
* Cow Milk, ** Goat Milk
Table 2. Distribution of isolates identied with MALDI-TOF MS (VITEK® MS)
Isolate Name Source
Boza Cheese Raw Milk Kefir
Enterococcus faecium - 43 83 1
Lactobacillus brevis 5 - - -
Lactobacillus plantarum - 3 1 3
Lactobacillus para plantarum - 3 - 2
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Investigation of Probiotic Features of ...
milk and cheese samples did not show any probiotic
properties. It has been reported that E. faecium obtained
from animal milk and cheese had good acidification and
strong bile salt tolerance in the previous studies [26,27].
However, these studies may be considered as incomplete
in terms of probiotic properties due to the lack of study
about the ability of colony formation in the intestinal
system.
In this study, the intestinal adhesion abilities, acid and
bile tolerance of the isolates were examined to determine
the probiotic properties of bacteria isolated from the
food samples. A similar study was conducted by Sanni [28]
for bacterial isolates from some regional food products
derived from grain, in which L. plantarum showed a good
and fast acid production capability. L. plantarum also
showed similar results in our study (Table 3).
A similar study was conducted by Banwo [29] for E.
faecium isolated from raw milk, and the technological and
Table 3. Acid-tolerant isolates and bile salt tolerances of isolates, hydrophobicity results
No Product Sample
No
Isolate
Code Microorganism
Acid
Tolerance
Bile Salt
Tolerance
Hydrophobicity
Ability
Viability (+/-) Viability (+/-) Hydrophobicity
(+/-)
1Boza 10 5 Lactobacillus brevis + + -
2Boza 3 78 Lactobacillus brevis + - -
3Boza 1 79 Lactobacillus brevis +++
4Boza 9 81 Lactobacillus brevis +++
5Boza 8 84 Lactobacillus brevis +++
6 Cheese 32 60B Enterococcus faecium + + -
7 Cheese 25 81B Enterococcus faecium + - -
8 Cheese 35 69B Enterococcus faecium + - -
9 Cheese 23 24B Enterococcus faecium + - -
10 Cheese 30 65B Enterococcus faecium + - -
11 Cheese 29 80B Enterococcus faecium + - -
12 Cheese 38 43C Enterococcus faecium + - -
13 Cheese 27 77C Enterococcus faecium + - -
14 Cheese 21 54C Enterococcus faecium + - -
15 Cheese 26 G76 Enterococcus faecium + - -
16 Cheese 31 G4 Enterococcus faecium + - -
17 Cheese 33 G37 Enterococcus faecium + - -
18 Cheese 46 E54 Lactobacillus paraplantarum + - -
19 Cheese 22 13B Lactobacillus paraplantarum + - -
20 Cheese 48 8C Lactobacillus paraplantarum + - -
21 Cheese 33 A21 Lactobacillus plantarum + - -
22 Raw milk 58 21B Enterococcus faecium + - -
23 Raw milk 98 70B Enterococcus faecium + - -
24 Raw milk 69 23B Enterococcus faecium + - -
25 Raw milk 72 8B Enterococcus faecium + - -
26 Raw milk 77 A19 Enterococcus faecium + - -
27 Raw milk 64 G11 Enterococcus faecium + - -
28 Raw milk 59 G37a Enterococcus faecium + - -
29 Raw milk 81 G1 Enterococcus faecium + - -
30 Raw milk 98 E14 Enterococcus faecium + - -
31 Raw milk 85 E75 Enterococcus faecium + - -
32 Raw milk 96 4C Lactobacillus plantarum + - -
33 Kefir 111 44C Lactobacillus plantarum +++
34 Kefir 112 74C Lactobacillus plantarum +++
35 Kefir 128 12C Lactobacillus plantarum +++
559
food safety characteristics of the species were investigated.
E. faecium species have been detected in respect to
resistance to bile salts and sensitive to antibiotics. In our
study, E. faecium species isolated from raw milk did not
show enough bile acid resistance.
In a study carried out by Gulel [30],
lactobacilli strains isolated from kefir
were able to resist both acid and bile
salts, but their hydrophobicity remained
low. In our study, the lactobacilli strains
isolated especially from kefir and boza
showed good hydrophobicity.
Probiotic bacteria must resist gastric
acidity and bile salts and adhere to the
epithelial surface of the intestinal mucosa.
These properties are fundamental criteria
for a bacterium to be a probiotic [31].
However, the relevant bacteria must be
tested for antibiotic resistance and anti-
biotic resistance genes to ensure the
safety for human consumption. In the
studies carried out by Sanni and other
researchers, the detection of antibiotic
resistance status of the microorganisms
seems to be missing [32-34]. It would be
useful to consider these criteria, which
should be examined in terms of food
safety, among the probiotic properties.
Thus, the probiotic character of examined
the microorganisms should be the end
result.
Acid tolerance is one of the most
important criteria for probiotic bacteria
as they are destroyed by the acidity of the
stomach [35]. Probiotic bacteria are more
resistant to stomach acidity than other
microorganisms and are usually exposed
to stomach acid with pH between 2.5
and 3.5 before arriving the colon. Acidic
conditions are one of the important
physiological challenges encountered by
probiotic bacteria [20]. In our study, 13 strains of the 16
Lactobacilli showed resistance to pH 2.5. Besides, 22 (17%)
of 126 E. faecium strains were able to show resistance
to pH 2.5. However, there are technological methods
recommended to analyze the probiotic bacteria for their
DOGAN, OZPINAR
Table 4. Antibiotic disc confirmat ion zones (mm) of the samples, antibiogram confirm ation and MIC (μg/ml) results
No CAZ
ZON CAZ CV CTX
ZON CTX CV CPD
ZON CPD CV CAZ CAZ
MIC
CAZ
CV MIC CTX CTX
MIC
CTX CV
MIC ESBL
3 16 18 24 25 23 23 R 32 >32/4 S ≤1 ≤0.5/4 -
4 - - - - - - ? - - S ≤1 ≤0.25/4 -
5 - - - - - - S ≤1 ≤0.25/4 S ≤1 ≤0.25/4 -
33 - - - - - - S ≤1 ≤0.25/4 S ≤1 ≤0.25/4 -
34 - - - - - - S ≤1 ≤0.25/4 ? - - -
35 18 18 15 15 18 18 S ≤1 ≤0.25/4 S ≤1 ≤0.25/4 -
CAZ: Ceftazidime, CTX: Cefotaxime, CPD: Cefpodoxime, CV: Clavulanate
The isolates were identi fied as ESBL (-)
Fig 1. Distribution of isolates exceeding stomach resistance
Fig 2. Distribution of bile-tolerant isolates
560
Investigation of Probiotic Features of ...
ability to pass through the stomach without being
destroyed. The most commonly used method is micro-
encapsulation. In principle, the powdered form of isolates
is covered with a suitable material enabling bacteria
to pass through the acidic environment of the stomach
without getting killed [36]. In a study by Mishra and Prasad [37],
three strains (43%) of seven lactobacilli were reported
to be resistant to pH 2.0 or 3.0. It is indicated that the
dierences in pH resistance of dierent species and even
of the same species are attributed to the dierences in
the multiplication stage of the bacteria [38].
The probiotic bacteria pass through the acidity of the
stomach and then come into the contact with bile [39].
Bile salt tolerance is another important criterion used in
the selection of probiotic bacteria [30]. Therefore, bacteria
to be used as probiotics need to be resistant to bile to
maintain their viability in the small intestine, a part of the
gastrointestinal tract [40]. The present study found that L.
brevis obtained from boza and L. plantarum obtained from
kefir showed resistance to bile salts. E. faecium obtained
from cheese and raw milk did not show enough resistance.
In particular, the earlier studies on L. brevis and L. plantarum
confirm the findings of our study. In the studies carried
out by Ronka [41], Ramos [42], and Golowczyc [43], L. brevis
and L. plantarum isolates exhibited good resistance to
bile. However, in another study, 86 of the 122 E. faecium
species isolated from traditional cheese samples (about
70%) were reported to be highly resistant to the medium
containing 0.3% bile. In addition, E. faecium was reported
to be more resistant to the harsh conditions of the
gastrointestinal tract than other probiotic bacteria [44].
An important criterion for the selection of probiotic
bacteria is their ability to colonize by attaching to the
epithelial surfaces on the intestinal mucosa. A positive
correlation has been observed between adhesion of
bacterial cells and bacterial cell surface hydrophobicity [45,46].
In our study, L. plantarum obtained from kefir and L.
plantarum obtained from boza showed
high hydrophobicity. The probiotic pro-
perties of L. plantarum isolated from
traditional Iranian dairy products and
L. brevis obtained from Brazilian origin
products were analyzed by Nejati [47]
and Ramos [42] respectively, and the
hydrophobicity abilities were found
high. These studies conform to our
findings on the high hydrophobicity
of L. brevis and L. plantarum isolates
obtained from dierent food samples [48].
According to the criteria established
by Food and Agriculture Organization
of the United Nations/World Health
Organization (FAO/WHO) [31], bacteria with
resistance to antibiotics and able to
transfer the antibiotic resistance genes
are considered unsafe for health and cannot be used as
probiotics [49]. Therefore, transfer of antibiotic resistance
genes by probiotics, especially to pathogenic bacteria is
the most important risk factor and needs to be controlled [50].
Earlier studies have shown that transfer of antibiotic
resistance genes to pathogenic bacteria from the
Lactobacillus species found in the intestinal ora may be
possible in limited numbers [51,52]. In our study, antibiotic
resistance and especially the presence of ESBL were not
observed in any bacterium. However, Lactobacillus strains
carrying the genetic vancomycin resistance gene may be
reliably used as probiotics, as no evidence has been shown
for the transfer of this gene to other strains [53].
In a study by Gulel [30], although the Lactobacillus strains
isolated from the kefir showed high resistance to nucleic
acid synthesis inhibitors and cytoplasmic membrane
inhibitors, a lower resistance to cell wall inhibitors and most
of the protein synthesis inhibitors was observed. In our
study, none of the isolated Lactobacillus strains displayed
antibiotic resistance to nucleic acid synthesis, cytoplasmic
membrane, and cell wall and protein synthesis inhibitors.
In a study carried out by Zheng [54], L. plantarum isolated
from kefir was susceptible to gentamicin, erythromycin,
and chloramphenicol inhibitors, whereas it showed
resistance to vancomycin.
In a study carried out by Forssten [55], the presence of
ESBL was determined by administering a probiotic blend
of Lactobacillus strains during antibiotic treatment, and
ESBL negative results were obtained. L. plantarum and L.
brevis strains isolated from kefir and boza yielded ESBL-
negative results in a similar manner.
Kefir and boza have been produced by the fermentation
of mixed cultures, including Lactobacilli species, could be
regarded as beneficial microorganisms [23,24]. Especially, the
in vivo studies on kefir have reported beneficial eects
on health [56-59]. The result of our study show that the
Fig 3. Distribution of isolates displaying high hydrophobicity
561
bacteria obtained from kefir and boza displayed probiotic
properties. This explains the beneficial eects of probiotic-
containing boza and kefir on health. However, the bacteria
from cheese and milk samples did not show enough
probiotic properties. These observations indicated that
kefir and boza consist of more bacteria with probiotic
bacteria as compared to cheese and milk.
The present study determined that L. brevis and L.
plantarum isolated from kefir and boza were able to
compansate the set probiotic criteria. The study indicates
that the probiotic bacteria may be obtained as an
alternative to industrial probiotics through non-GMO
(non-genetically modified organism) isolated from natural
fermented food products such as kefir and boza. Besides,
probiotics of Turkish origin were identified from the
bacteria isolated from kefir and boza samples.
ConfliCts of interest
The authors declare that there is no conict of interests
regarding the publication of this paper.
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