Received: 25/11/2024 Accepted: 20/01/2025 Published: 23/03/2025 1 of 7
https://doi.org/10.52973/rcfcv-e35579 RevistaCientíca,FCV-LUZ/Vol.XXXV
ABSTRACT
The aim of this study was to isolate and to identify lactic acid
bacteria from traditional cheeses of the Black See Region. Artvin
Şor, Giresun Tecen, Kargı Tulum, Ordu Kesik and Trabzon Telli
cheese were used as cheese samples of the Black Sea Region.
The number of lactic acid bacteria in traditional cheese of the
Black Sea Region were ranged from 4.62 ± 0.76 and to 7.87 ± 0.64
log cfu·g
-1
. Gram–positive and catalase–negative colonies were
evaluated as lactic acid bacteria based on the morphological
and biochemical properties. According to biochemical analysis
results, 39 lactic acid bacteria strains were identied by 16S
rDNA isolated from cheese samples. Based on the sequence
analysis, the indigenous lactic acid bacteria population was
identied as Enterococcus faecium (35.9%), as Levilactobacillus
brevis (12.8%), as Lactiplantibacillus plantarum (15.3%), as
Pediococcus acidilactici (7.6%), as Enterococcus durans (7.6%),
as Lacticaseibacillus paracasei (5.1%), as Lacticaseibacillus casei
(7.6%), as Leuconostoc mesenteroides (2.5%), as Leuconostoc lactis
(2.5%) and as Weissella cibaria (2.5%). Enterococcus spp. was
the dominant lactic acid bacteria in cheese sample. The present
ndings revealed that lactic acid bacteria populations varied
depending on cheese types in terms of cell counts and diversity.
Key words: Cheese; identication, lactic acid bacteria, PCR
RESUMEN
Este estudio tuvo como objetivo aislar e identicar bacterias
lácticas a partir de muestras de quesos tradicionales de la región
del Mar Negro. Se utilizaron quesos Artvin Şor, Giresun Tecen,
Kargı Tulum, Ordu Kesik y Trabzon Telli. El número de bacterias
lácticas osciló entre 4,62 ± 0,76 y 7,87 ± 0,64 log ufc·g
-1
. Las colonias
grampositivas y catalasa–negativas se evaluaron como bacterias
de ácido láctico en función de las propiedades morfológicas y
bioquímicas. Se identificaron 39 cepas de bacterias lácticas
mediante 16S rDNA aislado de muestras de queso. Con base en
el análisis de secuencia, la población de bacterias ácido lácticas
autóctonas se identicó como Enterococcus faecium (35,9%),
como Levilactobacillus brevis (12,8%), como Lactiplantibacillus
plantarum (15,3%), como Pediococcus acidilactici (7,6%), como
Enterococcus durans (7,6%), como Lacticaseibacillus paracasei
(5,1%), como Lacticaseibacillus casei (7,6%), como Leuconostoc
mesenteroides (2,5%), como Leuconostoc lactis (2,5%) y como
Weissella cibaria (2,5%), Enterococcus spp. fue la bacteria ácido
láctica dominante en la muestra de queso. Los presentes hallazgos
revelaron que las poblaciones de bacterias ácido lácticas variaban
dependiendo de los tipos de queso en términos de recuentos
celulares y diversidad.
Palabras clave: Queso; identicación; bacterias lácticas; PCR
Identication of lactic acid bacteria isolated from traditional cheeses
of the Black Sea Region
Identicación de bacterias lácticas aisladas de quesos tradicionales de la Región del Mar Negro
Selin Kalkan1 , Gökhan Özdemir1 , Kadriye Özcan2 , Emel Unal Turhan3*
1University of Giresun, Faculty of Engineering, Department of Food Engineering. Giresun, Türkiye.
2University of Giresun, Faculty of Engineering, Department of Genetics and Bioengineering. Giresun, Türkiye.
3University of Osmaniye Korkut Ata, Faculty of Kadirli Applied Sciences, Department of Food Technology. Osmaniye, Türkiye.
*Corresponding author: emelunalturhan@gmail.com
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INTRODUCTION
Cheese is a food product with high nutritional value, obtained
by pre–treating milk and usually fermenting it, and consumed
with pleasure all over the world. In addition, cheese is one of
the oldest fermented food products made by human [1]. France,
Netherlands, and Italy were famous for cheeses. Also, Turkey is
a very rich country in terms of cheese diversity [2]. Nowadays,
4,000 different variety of cheese are present in the world. Turkey
manufactures approximately 200 variety of cheese [3]. Although
the production methods of these cheese varieties are similar to
each other, their chemical, physical and sensory properties vary
depending on the milk, microflora or starter culture, environmental
factors such as climate and region. The Black Sea Region is one of
the regions of Turkey with rich cheese varieties. More than 30 types
of cheese are manufactured in this region. The most well–known
cheese types in the Black Sea Region are Civil Cheese, Kargi Tulum
Cheese, Imansiz Cheese, Kurchi Cheese, Minzi Cheese and Kolot
Cheese [4]. Kargı Tulum Cheese is a traditional cheese originating
from the Kargı district in Çorum province, Turkey. It is crafted using
a variety of milks, such as cow, sheep, or buffalo milk, depending
on local availability and preferences. It is typically made from
milk collected during the autumn season, which contributes to
its rich, high–fat content. Telli cheese is a traditional cheese from
the central districts of Trabzon and Artvin, as well as the Sürmene
and Akçaabat administrative districts of Trabzon in Turkey. It is
made primarily from cow’s milk. Telli cheese has thicker threads
compared to the thinner threads of Civil Cheese, which is also
sourer and salt–free in flavor. Artvin Şor cheese is made in the
Savsat district of Artvin and its vicinity. Its name derives from the
word Şor which means “bitter” in the region. This cheese is of a
dark yellow color, is very salty and has a bitter taste [4].
In recent years, due to the increasing demand of consumers
for natural products, interest in local cheese varieties has
increased [5]. It is possible that local cheese varieties will be
produced industrially and delivered much more and subsequently
become a demanded product in the world market [6]. However,
cheese industrially produced with starter cultures is different
from traditional cheese with spontaneous lactic microflora
(i.e. nonstarter lactic acid bacteria) regarding sensory quality
properties. Starter lactic acid bacteria (LAB) are deliberately added
to the milk for the production of acid during cheese processing.
However, nonstarter lactic acid bacteria or indigenous microflora
are inherently present in raw milk and play an important role in
the developing of cheese flavor [3, 7].
The presence of numerous enzymes and indigenous lactic acid
bacteria in milk improves quality properties of traditional cheeses.
Native LAB develop texture and flavour properties of cheeses with
microbiological and biochemical changes [7]. In industrial cheese
production, the main lactic acid bacteria utilized as starter cultures
are Lactobacilus casei, Lactobacillus helveticus and Lactobacillus
delbrueckii subsp. bulgaricus [8]. In order to produce traditional
cheese industrially, it is essential to determine the microflora of
the local cheese. Additionally, the characterization of the dominant
microflora provides whether it is possible to use as a starter culture
[9]. Recently, molecular methods have been utilized to identify
lactic acid bacteria. Especially, the sequences of the gene 16S
of the ribosomal RNA is highly effective method to describe the
phylogenetic degree of relatedness among bacteria. Hence, 16S
rRNA sequence analysis has been progressively performed to
characterize the bacterial diversity of various cheeses [10].
The dairy industry works with the limited variety of starter
cultures. However, there is a need for indigenous LAB strains as
starters to produce more flavorful cheeses [11]. A few studies
have reported to determine indigenous lactic acid bacteria in the
traditional cheese of the black sea region. This research aims to
identify lactic acid bacteria in the traditional cheese of the black
sea region by using PCR based molecular techniques.
MATERIALS AND METHODS
Cheese samples
Cheese samples were obtained from Black Sea Region in Turkey.
Traditional Turkish cheeses of Black Sea Region used in this study
are Telli cheese (TP) from Tonya/Trabzon, Şor cheese (AŞ) from
Artvin, Tecen cheese (TC) from Giresun, Kargı Tulum cheese (KT)
from Kargı/Çorum and Kesik cheese (OK) from Ordu.
Isolation and enumeration of lactic acid bacteria in traditional
cheeses
Cheese sample (25 g) was diluted with 225 mL of peptone
water (Sigma) and then homogenized with stomacher for 2 min
at 10 strokes per second. Serial dilution method is used for the
enumeration of lactic acid bacteria. Using a 1/10 dilution rate, the
homogenate was diluted to a dilution level of 10
7
. By taking 0.1 mL
from the appropriate dilutions, the homogenate was cultivated in
petri dishes containing the selective medium. The two selective
media were utilized for enumeration and isolation of bacteria. LAB
with mainly lactococci are grown on M17 agar acc. to TERZAGHI
(M17 Agar, Merck, Germany) at 30°C for 48–72 h under anaerobic
conditions; LAB with mainly lactobacilli are grown on de MAN,
ROGOSA and SHARPE (MRS) Agar (Merck, Germany) at 37°C for
48–72 h under anaerobic conditions. For anaerobic conditions,
anaerobic jars and anaerocult A were applied. Following incubation,
the colonies grown on M17 and MRS plates were enumerated.
From each cheese sample, approx. Ten colonies with distinct
morphological characteristics were selected from MRS and M17
agar plates and subsequently transferred to fresh plates for further
purication. The assessment of Gram–positive and catalase–
negative colonies was performed in a medium containing 6.5%
and 18% NaCl and at temperatures of 10 and 45°C, at pH 4.4
and pH 9.6. Furthermore, CO
2
production as a result of glucose
metabolism of the isolates and arrangement of cells (tetrad form,
chain form) was assessed. Gram–positive and catalase negative
pure isolates were selected from all cheese samples, and they were
stored at -20°C (Arçelik, 4252EY, Türkiye) with glycerol (Merck,
Darmstadt, Germany) stocks [7, 12, 13].
Extraction of bacterial DNA from pure isolates of cheese samples
Bacterial DNA from pure isolates was extracted using method
described in previous studies [7, 14, 15]. 2 mL overnight culture were
obtained for total DNA extraction. Microbial cells were centrifugated
(Hermle Z326K, Germany) for 5 min at 4,000 g at 4°C. Supernatant
was poured and then the pellets were resuspended in 500 μL of the
lysis buffer. After incubation (Nuve ES 120 cooled incubator, Türkiye)
for 10 min at 65°C, 150 μL potassium acetate was added. The mixture
Lactic acid bacteria in traditional cheeses / Kalkan et al._______________________________________________________________________________ _________________________________________________________________________________________________Revista Cientica, FCV-LUZ / Vol.XXXV
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was centrifugated for 5 min at 4°C and at 12,000 g and supernatant
was transferred into sterile Eppendorf tube by adding isopropyl alcohol
at equal volume. The mixture was centrifugated (Hermle Z326K,
Germany) for 2 min at 4°C and at 12,000 g and then supernatant
was discarded. The pellets were suspended in ethanol (70%, V/V)
and this mixture was centrifugated (Hermle Z326K, Germany) for 1
min at 4°C and at 12,000 g. The pellet washed in ice–cold 70% ethyl
alcohol. DNA was dried by vacuum centrifugation and resuspended in
50μL ultrapure water. The PCR products were electrophoresed (MS
Major Science, Mini 300, Taiwan) on a 1.5% agarose gel to examine
the excepted size and to separate amplication products. DNA was
stored at -20°C (Arçelik, 4252EY, Türkiye) for following trials.
16S rDNA sequence analysis
This research utilized two universal primers: 27F
(5′–CCGCGGCTGCTGGCACGTA–3′) and 1492R (5′–
GTGCGGGCCCCCGTCAATT–3′). For reaction, the mixture of 2µL
dNTP (10 pM), 2 µL primer (10 pM), 3 µL MgCl2, 10 µL10X tampon,
0.25 µL Taq polymerase (Go Taq Hot Start Polymerase) was
suspended with distilled pure water at 50 µL total volume. After an
initial denaturation was performed at 94°C for 5 min., the products
were amplied (PCR–BioRad T100 Thermal Cycler, USA) through 35
cycles. Each cycle was denaturation at 94°C for 30 s, annealing at
52°C for 40 s, elongation at 72°C for 90 s. Lastly, the nal extension
step was performed at 72°C for 5 min. The resulting PCR products
were then sent to BMLabosis (Ankara, Türkiye) in Ankara for the
process of sequencing. The sequences were compared with those
in the NCBI GenBank database utilizing the BLAST tool [7, 14, 15].
Statistical analysis
Statistical analyses were conducted utilizing Windows SPPS
20.0 software statistical package program (SPSS Inc., Chicago,
IL, USA) according to the randomized block experimental plan.
The data were evaluated using one–way analysis of variance and
signicance differences were determined according to Duncan’s
multiple comparison test.
RESULTS AND DISCUSSION
Number of lactic acid bacteria in traditional cheeses
TABLE I gives the mean quantities of LAB in cheese samples.
The numbers of LAB on MRS and M17 plates varied from 4.62 and
7.87, and from 4.89 to 8.80 log cfu·g-1, respectively.
The lowest LAB counts belong to Ordu Kesik cheese samples,
while the highest LAB counts belong to Trabzon Telli cheese
samples. In general, LAB counts in Kesik cheese and Telli cheese
was statistically different (P<0.05) from other cheese while
difference among LAB counts in Şor cheese, Tecen cheese and
Kargı Tulum cheese was found to be insignicant (P>0.05) except
Kargı Tulum cheese sample in M17 plate.
In line with the present results, previous researchers stated that
traditional cheeses contain lactic acid bacteria counts at wide range
from 4 and to 9 log cfu·g
-1
[3, 7, 16]. Various researchers highlighted
those differences in the number of LAB may result from production
methods, the type of milk, storage period, and regional factors [2, 16,
17]. For example, the number of LAB in Telli cheese is found quite
high in this study since Trabzon Telli cheese is primarily produced in
farms under primitive conditions and usually based on spontaneous
fermentation [18]. In brief, spontaneous fermentation, a long
period of ripening period and traditional production techniques
are responsible for numerous LAB in cheese [18].
Isolation and identication of lactic acid bacteria in traditional
cheeses
A total of 147 strains were isolated, with 75 strains from MRS
agar and 72 strains from M17 agar. These strains were subjected to
morphological and biochemical tests. Lactic acid bacteria isolates
were morphologically examined using a light microscope following
gram–staining and catalase tests.
The morphological and biochemical properties of lactic acid
bacteria colonies were examined before DNA extraction. The
Gram–positive and catalase negative colonies were divided into
three groups consisting of cocci, bacilli or coccobacilli. In the
M17 plates, the cocci form (61.90%), bacilli form (28.58%) and
coccobacilli form (9.52%) were observed at different proportions.
In the MRS plates, bacilli and cocci colonies were 55.56% and
38.88% of the total lactic acid bacteria colonies, respectively
while coccobacilli represented 5.56% of all isolates. The cocci
mesophilic colonies were signicantly higher in Tecen and Şor
cheese according to other cheese samples and bacilli colonies
were higher in Kargı Tulum cheese.
According to biochemical test results, total 39 strains including
8 strains from Kesik cheese, 5 strains from Şor cheese, 10 strains
from Tecen cheese, 8 strains from Kargı Tulum cheese and 8 strains
from Telli cheese were selected for further identication. These
39 isolates were dened as lactic acid bacteria according to the
results of PCR. FIG. 1 presents the gel images obtained after
the PCR analysis of the samples. These 39 strains include 14
Enterococcus faecium (35.9%), 5 Levilactobacillus brevis (12.8%),
6 Lactiplantibacillus plantarum (15.3%), 3 Pediococcus acidilactici
(7.6%), 3 Enterococcus durans (7.6%), 2 Lacticaseibacillus
paracasei (5.1%), 3 Lacticaseibacillus casei (7.6%), 1 Leuconostoc
mesenteroides (2.5%), 1 Leuconostoc lactis (2.5%) and 1 Weissella
cibaria (2.5%).
TABLE II displays identication of LAB isolates from cheese
samples. As a result of sequence analysis, the autochthonous
LAB strains was defined as Enterococcus spp. (43.58%),
Lactiplantibacillus spp. (15.38%), Lacticaseibacillus spp.
TABLE I
The numbers of LAB in cheese samples (log CFU·g-1)
Samples MRS agar M17 agar.
TP 7.87 ± 0.64c8.80 ± 0.59d
AŞ 6.28 ± 0.25b7.95 ± 2.93c
TC 7.14 ± 0.12bc 7.35 ± 0.61c
KT 6.38 ± 0.21b5.72 ± 0.03b
OK 4.62 ± 0.76a4.89 ± 0.40a
a–b:DierentsuperscriptsinthesamecolumnindicateasignicantdierenceatP<0.05.,
TP:TellicheesefromTonya/Trabzon,AŞ:ŞorcheesefromArtvin,TC:Tecencheesefrom
Giresun,KT:KargıTulumcheesefromKargı/ÇorumandOK:KesikcheesefromOrdu
Lactic acid bacteria in traditional cheeses / Kalkan et al._______________________________________________________________________________
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(12.82%), Levilactobacillus spp. (12.82%), and Pediococcus spp.
(7.69%), Leuconostoc mesenteroides (5.12%), as Weissella cibaria
(2.56%). Out of the 17 Enteroccus spp., 14 were classied as
Enterococcus faecium, and 3 as Enterococcus durans. Out of the 5
Lacticaseibacillus spp., 3 were classied as Lacticaseibacillus casei
and 2 as Lacticaseibacillus paracasei. Out of the 2 Lecuconostos
spp. were classied as 1 Leuconostoc lactis and as 1 Leuconostoc
mesenteroides. Out of the other 15 strains were classied as 6
Lactiplantibacillus plantarum, as 5 Levilactobacillus brevis, as 3
Pediococcus acidilactici, and as 1 Weissella cibaria.
In addition to the counts of the microbial populations, the
distribution of their morphological types (cocci, bacilli and
coccobacilli colonies) and the distribution of their species may
vary depending on environmental and genetic parameters [10]. The
Enterococcus spp. (47.2%) were the predominant group which is in
line with the study results of Aktaş and Erdoğan [19] Enterococccus
strains show tolerance to salt concentration and acidic conditions.
The presence of Enterococcus strains in cheese resulted from
insufcient hygiene and sanitation practices during the handling of
raw milk or processing equipment. Additionally, Enterococcus strains
are present in milk because of contamination from the external
surface of dairy animals, unhygienic dairy equipment, dairy storage
tanks, or water sources contaminated with feces. These strains
affect quality properties of traditional cheeses such as mainly flavor
and texture [7]. In line with the present results, Demirci et al. [20]
stated that Enterococcus spp. is the dominant species in traditional
cheese. Similarly, in another study, it was reported that Enterococcus
isolates from Ezine cheese were mostly E. faecium (64.3%) [11].
Enterococcus strains are part of the natural microflora in cheese
produced from raw milk and induce the ripening process and improve
sensory quality. In addition to this, the presence of enterococci in
dairy products results from the process of milking and storing [21].
FIGURE 2, gives distribution of lactic acid bacteria present in
cheese samples. In the present study the greatest prevalence
among genera was of Enterococcus spp. Particularly, Enterococcus
feacium is the common strain identied for all cheese samples.
Additionally, Şor cheese and Tecen cheese contain higher amount
of E. feacium according to the other cheeses. Lactiplantibacillus
plantarum was the second most prevalent lactic acid bacteria
FIGURE 1. Gel images of LAB isolates by16S rDNA PCR
TABLE II
Identication of lactic acid bacteria in cheese samples
Cheese
samples Isolate Similarity based on
16S rDNA Similarity (%)
Kesikcheese
OK–M17-1 Enterococcus faecium 100
OK–M17-5 Levilactobacillus brevis 99
OK–M17-9 Leuconostoc lactis 99
OK–M17-13 Lactiplantibacillus plantarum 100
OK–M17-15 Leuconostoc mesenteroides 97
OK–MRS-2 Lactiplantibacillus plantarum 99
OK–MRS-4 Weissella cibaria 100
OK–MRS-14 Lactiplantibacillus plantarum 100
Şorcheese
AŞ–M17-2 Pediococcus acidilactici 100
AŞ–M17-5 Pediococcus acidilactici 99
AŞ–M17-11 Enterococcus faecium 98
AŞ–M17-15 Enterococcus faecium 99
AŞ–MRS-7 Pediococcus acidilactici 99
Tecen cheese
TC–M17-3 Enterococcus faecium 99
TC–M17-5 Enterococcus faecium 99
TC–M17-7 Enterococcus faecium 99
TC–M17-8 Enterococcus faecium 99
TC–M17-11 Lactiplantibacillus plantarum 99
TC–MRS-2 Enterococcus faecium 99
TC–MRS-3 Enterococcus faecium 99
TC–MRS-7 Enterococcus faecium 99
TC–MRS-9 Enterococcus durans 100
TC–MRS-15 Enteroccoccus faecium 100
KargıTulum
cheese
KT–M17-2 Levilactobacillus brevis 100
KT–M17-8 Levilactobacillus brevis 100
KT–M17-10 Levilactobacillus brevis 99
KT–M17-12 Enteroccocus faecium 99
KT–MRS-3 Lacticaseibacillus casei 99.8
KT–MRS-6 Lactiplantibacillus plantarum 100
KT–MRS-13 Levilactobacillus brevis 100
KT–MRS-14 Lactiplantibacillus plantarum 99
Tellicheese
TP–MRS-5 Lacticaseibacillus paracasei 100
TP–MRS-6 Lacticaseibacillus casei 99.57
TP–MRS-11 Lacticaseibacillus paracasei 100
TP–MRS-12 Lacticaseibacillus casei 100
TP–MRS-15 Enterococcus faecium 100
TP–M17-3 Enterococcus durans 99
TP–M17-7 Enterococcus durans 98
TP–M17-8 Enterococcus faecium 98
Lactic acid bacteria in traditional cheeses / Kalkan et al._______________________________________________________________________________ _________________________________________________________________________________________________Revista Cientica, FCV-LUZ / Vol.XXXV
5 of 7
in cheese samples of the present study. The highest diversity of
lactic acid bacteria species was veried in the Ordu Kesik cheese
samples with 6 different species including Enterococcus faecium,
Levilactobacillus brevis, Leuconostoc lactis, Lactiplantibacillus
plantarum, Leuconostoc mesenteroides and Weissella cibaria.
Furthermore, Weissella cibari and Leuconostoc lactis were identied
in only Ordu Kesik cheese. Pediococcus pentosaceus (FFH20) was
only isolated from the Artvin Şor cheese samples. Telli cheese is
rich in Lacticaseibacillus spp. compared with other cheese samples.
Similar to the results in TABLE I, and FIG. 2 shows that in terms
of similarity in the distribution of LAB, Şor cheese, Tecen cheese
and Kargı Tulum cheese form a similar group while Telli cheese
and Kesik cheese form another group. In this study, Enteroccus
durans was identied in Tecen cheese and Telli cheese.
Medeiros et al. [10] highlighted that high lactic lactic bacteria
counts indicate microbial richness. On the contrary, in this study,
kesik cheese with low LAB counts had much more microbial
richness involving a large variety of different lactic acid bacteria
species. As known from literature reports, microorganisms have
antagonistic activity and dominant microflora may inhibit other
species of strains (Rençber et al. [7]). This explains the high variety
of LAB and low variety of LAB in Ordu kesik cheese and Artvin Şor
cheese, respectively.
Similarly, previous researchers reported that Lactiplantibacillus
spp. was second dominant microflora in Tulum cheeses [7, 22].
They have great potential to utilize as starter culture since
Lactiplantibacillus species are resistant to low pH and high amounts
of salt in cheese [11]. Lactobacillus was another dominant species
in cheese manufactured from raw milk due to their growth under
hard selective conditions and proteolytic activities [23].
Chourasia et al. [24] stated that Enterococcus durans was the
main LAB in chhurpi cheese. The excessive use of antibiotics in
agricultural and clinical practices has boosted the prevalence of
antibiotic–resistant bacteria in food products. Antibiotic–resistant
enterococci are among the most crucial microorganisms associated
with infections [24].
The distribution of LAB genus of traditional cheeses was
not homogeneous in all locations or regions. The deciency of
standardization of the traditional cheeses manufactured in all
regions causes heterogeneity in distributions of LAB genera. This
heterogeneity provided specic sensory characteristics of this
traditional food product [10].
Diversity of LAB strains resulted from specic to traditional
cheese, commonly processed with raw milk. Raw milk or non–
pasteurized milk is abundant in all lactic acid bacteria including
non–starter lactic acid bacteria. The main source of the cheese
microflora is considered as the environment, water, the mammary
gland, processors of dairy products or other materials during the
milk manufacturing process [10]. Traditional cheeses involve
indigenous lactic acid bacteria populations which cause unique
texture and aroma of cheese. Pasteurization damages the natural
microflora of milk and dairy products. Hence, traditional starter
cultures are added into milk after the pasteurization process [16].
Nonstarter lactic acid bacteria are naturally present in raw milk
microflora. Particularly they improve aroma and flavor in long–term
ripened cheese [21].
CONCLUSIONS
The present results indicated a great and important diversity
of LAB in the traditional cheeses of the Black Sea Region. The
sequencing of the 16S rRNA gene was a highly effective technique
for the identication of lactic acid bacteria in cheeses. The most
prevalent genera in cheese samples were Enterococcus spp. and
Lactiplantibacillus spp. In general, the distribution of the LAB
species in cheese samples was different from each other. In the
next study, the influence of identied lactic acid bacteria on the
technological properties of cheeses and the potential for use as
a starter culture may be focused on.
ACKNOWLEDGEMENTS
To Giresun Üniversitesi Bilimsel Araştırma Projeleri (BAP)
Komisyon Başkanlığına (Proje No: FEN–BAP–C-281119-78).
Financial Support
This study was supported by Giresun Üniversitesi Bilimsel
Araştırma Projeleri (BAP) Komisyon Başkanlığına (Project
No: FEN–BAP–C-281119-78).
Conflict of Interest
The authors declared that there is no conflict of interest.
Ethics approval
Not required. Any of the author conducted no human or animal
studies in this article.
Consent for publication
Not required.
Compliance with ethical standards
There are no studies with human or animal subjects in this article
OK AŞ TC KT TP
0
10
20
30
40
50
60
70
80
90
100
Lacticaseibacillus casei
Lacticaseibacillus paracasei
Enterococcus durans
Pediococcus acidilactici
Weissella cibaria
Leuconostoc mesenteroides
Lactiplantibacillus plantarum
Leuconostoc lactis
Levilactobacillus brevis
Enterococcus faecium
FIGURE 2. Distribution of lactic acid bacteria present in cheese samples
Lactic acid bacteria in traditional cheeses / Kalkan et al._______________________________________________________________________________
6 of 7 7 of 7
Author Contributions
S.K.: Conceptualization, supervision, methodology, statistical
analysis, writing–original draft, review & editing, K.Ö.: Methodology,
data curation and analysis, writing–original draft, review & editing.
G.Ö.: Review, writing–original draft, data curation and analysis E.U.T:
Methodology, supervision, writing–original draft, review & editing
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