LAB counts of pickles and isolation

LAB counts of the pickles were ranged between < 1 and 6.81 log CFU/g (Table S1). LAB isolates were not obtained from samples F, I and J.

Probiotic properties of LAB

In the study, 114 isolates exhibiting Gram-positive and catalase-negative characteristics were initially selected as potential LAB. Subsequently, the probiotic properties of these isolates were investigated. In the study, 18 isolates were identified at the molecular level.

Tolerance to different temperatures, pH, NaCl, bile salts and phenol

Within the scope of this study, the growth characteristics of the isolates were studied at three different temperatures (10, 25 and 45 °C) (Table 1). Most of the isolates showed better growth capabilities at 25 °C, while no one could grow at 10 and 45 °C (Table 1). Similarly, in a study, the viability of fifteen LAB isolated from traditional pickles specific to India was tested at 15, 30 and 45 °C, and all isolates grew at 15 and 30 °C, but only seven isolates maintained their viability at 45 °C [15]. The findings revealed that, among the temperatures tested, the optimal temperature for probiotic growth was 25 °C. However, probiotic microorganisms should remain alive in gastrointestinal system (GIT) to confer health benefits on the host. For the survival of probiotics, tolerance to the stress conditions in the GIT, such as the presence of low pH, phenol, NaCl, and bile acids is crucial. Probiotic viability is typically affected at pH values below 2.5. While this is crucial for eliminating harmful microorganisms, it can be lethal to probiotics. Table 1 presents the results of acid tolerance of isolates at pH 2, 3, and 4. The pH of the environment is one of the factors that significantly affects the growth of microorganisms [43]. Researchers indicated that aciduric members, like L. acidophilus, are sensitive at pH 2.0 and below, and that they typically cannot survive in low pH environments. However, to qualify as a probiotic, a bacteria must be able to withstand high levels of acidity and sustain a high count for 2 to 3 h while in the stomach. In this study, none of them could survive at pH 2 while 65 isolates could survive at pH 3 and 91 could survive at pH 4. However, acid adaptation can be implemented to facilitate the growth of these isolates under low pH conditions. Salt tolerance allows LAB to start fermentation and acid production quickly and has a suppressive effect on competing microbiota that are intolerant to salt [44]. The results showed that all isolates can survive in medium containing 1.5% (v/v) NaCl. However, 73 isolates could tolerate to NaCl at 10% (v/v) (Table 1). LAB has varying degrees of bile salt hydrolysis (BSH) activity, which protects against bile acid toxicity [45]. The tolerance to bile salt of the isolates was determined in the presence of 0.3 and 1% bile salts. Although the growth of the isolates was faster in medium containing 0.3% (v/v) bile salt, most isolates could tolerate 0.3 and 1% bile salt concentrations (Table 1). Acid and bile tolerances of isolates varied significantly due to species specificity [46, 47]. On the other hand, phenols can be formed because of the deamination of amino acids ingested with food or produced in the body by bacteria in the digestive system. These phenolic compounds may have an inhibitory effect on Lactobacillus spp. In this study, 64 isolates could survive in phenol (0.4%) presence for 24 h (Table 1). In a study carried by Boricha et al. [48], the counts of LAB isolated from various fermented foods were detected in the range of 6.2–7.7 log CFU/mL, 5.2–6.7 log CFU/mL, 5.8–7.1 log CFU/mL and 6.1–7.5 log CFU/mL in media having pH 3; 4% (w/v) bile salt; 6% (w/v) NaCl and 0.6% (w/v) phenol, respectively. The result of the study carried by Rao et al. [16] showed that L. plantarum AT4 and L. plantarum AT282 isolated from traditional Chinese pickles could survive in the presence of 0.1, 0.3, and 0.5% bile salt, while the number of L. plantarum AT282 decreased in the presence of 0.5% bile salt. Similarly, in this study, three isolates could tolerate 0.3 and 1% bile salt concentrations although the growth of the isolates was faster in medium containing 0.3% (w/v) bile salt. This result is significant because bile tolerance is also an essential factor for probiotic to survive, grow, and exert their action within the GIT. Moreover, the bile salts in the body usually do not exceed 0.3%. Nevertheless, probiotics must be able to survive in the stomach and intestine. A study conducted by Lin et al. [12], L. plantarum AR113, AR501 and P. pentosaceus AR243, which were isolated from Chinese traditional pickle, exhibited promising probiotic activities by tolerating low pH (3.0), high bile salt (1.1%) and high osmotic pressure (8.0%). High bile salt tolerances were also reported for L. brevis B13-2 isolated from kimchi [49] and L. plantarum SK 1305 from Korean green chili pickled pepper [14]. These findings, which are parallel with the current study, confirming that LAB isolated from pickle can tolerate stress conditions.

Table 1 Survival of isolates at different conditionsTolerance to pepsin and pancreatin

The tolerance of isolates to pancreatin and pepsin is another criterion for predicting their survival in the harsh conditions of the GIT. Therefore, microorganisms used as probiotic must remain alive during their passage through the digestive system. The results showed that most of the isolates were alive in the presence of pepsin (81) and pancreatin (78) after 4 h and 6 h incubation, respectively (Table 1). Tokatlı et al. [30] stated that LAB counts, which isolated from various pickles, were more than 6 log units for 17 isolates, while four isolates completely lost their viability in the presence of pepsin after 4 h of incubation. It was also detected that twelve isolates could be alive in the presence of pancreatin with decreased numbers.

Antimicrobial activity

The antimicrobial action of LAB isolates is attributed to the production of bacteriocins, organic acids, and metabolites during growth [50]. Based on the isolates and test microorganisms used, 86 isolates exhibited antimicrobial activity, with a range of inhibition zones. A total of 20 isolates demonstrated a diverse range of antimicrobial activity, affecting four distinct test microorganisms. Four isolates demonstrated antimicrobial activity against B. cereus No. 8, 33 isolates against S. Typhimurium NRRL B-4420, and the highest number of isolates (65 isolates) against S. aureus 6538P. However, the isolates did not demonstrate any activity against L. monocytogenes Scott A (Table 2). Researchers found that some isolates obtained from traditional Indian pickles inhibited the growth of pathogens, including E. coli, Shigella dysenteriae, and S. aureus. The antimicrobial activity was not observed for S. dysenteriae and S. aureus, and the highest antimicrobial activity was observed against B. cereus (16.36 mm) by L. plantarum (PKL-21) [15]. Cizeikiene et al. [51] employed the agar diffusion method to ascertain the antimicrobial activity of the isolates. The isolate P. pentosaceus KTU05-10 demonstrated the highest activity against P. cepacia 7.2 (zone diameter 27.5 mm), yet none of the isolates exhibited antimicrobial activity against S. aureus ATCC 25923 and L. monocytogenes. These studies have also demonstrated that antimicrobial activity varies between isolates and species.

Table 2 Antimicrobial activity and proteolytic enzyme activity of isolates

The results of the study indicate that the majority of the strains exhibited antimicrobial activity against at least three test cultures, demonstrating that they have important bioprotective potential in terms of food safety and human health.

Determination of enzyme activityProteolytic activity

In consideration of the preceding analysis, the isolates exhibiting similar characteristics were grouped, and 39 representative isolates were selected for testing of their proteolytic activities. As documented in the literature, LAB are reported to exhibit relatively low proteolytic activity. The proteolytic activity of the isolates exhibited considerable variation, with values ranging from 1.8 µg/mL tyrosine to 126.2 µg/mL tyrosine (Table 2). In a study conducted by Monika et al. [15], protease activity was detected in 14 out of 15 LAB isolates, with specific activities ranging from 0.1 to 3.64 U/mg protein. Nevertheless, in the present study, the proteolytic activity of 18 isolates exceeded 50 µg/mL tyrosine. The influence of isolates with high proteolytic activity on product flavor is a notable consideration. Hyperactive proteolytic enzymes have been observed to stimulate the formation of a bitter flavor and the production of other unfavorable metabolites, which may have implications for product quality and consumer acceptability.

β-Galactosidase activity

Lactose intolerance is primarily attributable to a deficiency of β-galactosidase, an enzyme that hydrolyses lactose into glucose and galactose. Nevertheless, probiotic isolates have been demonstrated to confer significant benefits to individuals with lactose intolerance by facilitating the metabolism of lactose [52]. In this study, four isolates demonstrated β-galactosidase activity (data not shown). Boricha et al. [48] observed that L. pentosus and L. rhamnosus isolates obtained from various pickles exhibited varying levels of β-galactosidase activity. Additionally, Wang et al. [53] documented the highest β-galactosidase activity for L. paracasei F08 and L. acidophilus C11, which were isolated from sauerkraut produced in Taiwan. Probiotic isolates are capable of metabolising lactose, which is beneficial for individuals with lactose intolerance [54].

Antibiotic susceptibility

A considerable number of LABs have been observed to exhibit resistance to antibiotics. These resistance attributes are frequently intrinsic and non-transmissible. While LAB used in the production of fermented and probiotic foods are typically regarded as safe microorganisms, the potential for transfer of antibiotic resistance genes to pathogens or other bacteria in the microbiota represents a significant risk [12]. Consequently, the antibiotic susceptibility of the isolates was ascertained through the disc diffusion method, employing AMP, C, E, CN, and K. All isolates exhibited resistance to kanamycin, while six isolates (GL1, GL12, GL13, HL1, HL29, and HL32) demonstrated susceptibility to gentamicin. Most of the isolates displayed susceptibility to AMP, E, and C. However, eight isolates (BL5, BL13, Dl6, DL8, HL7, HL13, HL22, and HL31) exhibited resistance to the antibiotics (Table 3). In a study, fifteen isolates obtained from traditional pickles of India were found to be susceptible to all antibiotics (penicillin, erythromycin, vancomycin, teicoplanin, clindamycin, ofloxacin, azithromycin and tetracycline) tested, except for vancomycin and teicoplanin [15]. Boricha et al. [48] observed that isolates derived from disparate pickles exhibited heightened susceptibility to a broader array of β-lactam antibiotics, though this was not the case for amikacin, methicillin, teicoplanin, cefpirome, ceftazidime, cefepime, norfloxacin, ciprofloxacin, ceftriaxone, kanamycin, or streptomycin. Probiotics must be safe microorganisms. It is imperative to ascertain that LAB employed in the food industry do not harbour antibiotic resistance genes, which may potentially pose a risk of transferring the resistance gene to other microorganisms [55]. However, minor discrepancies in comparison with the present study may be attributed to variations in isolate and species.

Table 3 Antibiotic susceptibility of isolates (mm)Haemolytic activity

As a safety requirement for the selection of a probiotic strain, the isolate must not have haemolytic activity [56]. The results demonstrated that the LAB isolates did not exhibit α and β haemolytic activity (data not shown). Similar results were obtained for E. faecalis, L. plantarum, L. mesenteroides, L. lactis, P. pentosaceus and Enterococcus spp. isolated from traditional Indian pickles [15], L. plantarum, P. ethanolidurans and L. brevis isolated from traditional Iranian pickles [57] and W. cibaria, B. subtilis and B. tequilensis isolated from South Indian tomato pickles [58]. Conversely, it was reported that the most of isolates (Bacillus and Enterobacter spp.) isolated from traditional Chinese pickles demonstrate haemolytic activity [59]. This finding indicates that these isolates cannot be considered a safe probiotic isolate. The isolates obtained from pickles did not exhibit heamolytic activity in our study, which is significant since it supports their potential for safe use as probiotics.

Genotypic identification of presumptive probiotic isolates

Within the scope of this study, the isolates firstly examined for probiotic properties and then presumptive probiotics (AL4, AL14, BL1, BL8, BL13, BL16, CL7, CL17, DL2, DL6, DL9, GL3, GL12, HL4, HL13, HL14, HL24, and HL31) were identified at the molecular level (Table 4). A phylogenetic tree based on 16S rRNA gene sequences was constructed using the neighbor-joining and Tamura-Nei methods with bootstrap test (1000 replicates) to understand the relationship between the isolates. The results revealed that these isolates belong to five different species: L. plantarum, L. pentosus, L. brevis, P. parvulus and L. parabuchneri (Table 4). Among them, L. plantarum and P. parvulus were dominant species. According to data obtained through molecular identification results, 16S rRNA gene sequence studies were sufficient to distinguish between isolates of L. brevis, L. parabuncheri, and P. parvulus but not L. plantarum and L. pentosus (Fig. S2).

Table 4 Molecular identification results of the isolatesPresumptive probiotic counts in cucumber pickles during fermentation and storage

Formulation stability and probiotic cell viability in the product during storage are essential criteria for a commercial probiotic product. Thus, presumptive probiotic counts were performed on the samples during fermentation and storage (Figs. 1 and 2).

Fig. 1

Number of presumptive probiotics in cucumber pickles during fermentation and storage

Fig. 2

Number of yeasts in cucumber pickles during fermentation and storage

During fermentation, presumptive probiotic counts ranged between 6.50 log CFU/g and 8.24 log CFU/g. At the beginning of fermentation (day 0), the LAB count was in the range of 6.50–7.42 log CFU/g for the samples enriched with presumptive probiotics, while it was 3.40 log CFU/g for NC. In the following days, an increase in the presumptive probiotic count was observed, and the highest presumptive probiotic count (7.85 log CFU/g) was determined on the third day of fermentation in PS1, produced with the addition of L. plantarum (P < 0.05). Also, on the third day of fermentation, the presumptive probiotic count in the positive control (PC) fermented with L. plantarum 299v was 7.32 log CFU/g (P > 0.05). After 15 days of fermentation, PS3 had the highest presumptive probiotic count, with 7.51 log CFU/g. However, there was no statistically significant difference between PS3 and PS1, PS2, PS4 or PC (P > 0.05). The LAB count in the NC increased on the first day of fermentation, then began to decrease on the 5th day of fermentation, reaching 5.76 log CFU/g at the end of fermentation (P < 0.05). In addition, the lowest presumptive probiotic count (6.58 log CFU/g) was detected in the PS5, produced by mixed culture (P < 0.05) (Fig. 1). Due to competition for food sources and a quicker pH decrease, probiotic cells lose their viability, which explains why mixed culture pickles have lower numbers of probiotic cells than single culture pickles [23, 60]. This study shows that probiotics added to pickle samples can maintain their viability above a certain level (> 6 log CFU/g) until the end of fermentation. Therefore, the pickles meet the " > 6 log CFU/g live probiotic cell criterion" sought in probiotic products. Besides, the LAB count in NC does not meet the criteria needed for the product to be considered as probiotic (which is also unclear whether it is probiotic or not). In a similar study conducted by Alp and Kuleasan [61], five different groups of garlic (Allium sativum) pickles were produced adopting the Turkish-style fermentation using monocultures or mixed cultures containing four different probiotic strains at a level of 1.5 × 108 CFU/mL (L. plantarum DA100, L. fermentum DA134, L. coryniformis DA256, Leuconostoc lactis DA268). They found that LAB counts were below the detection limit in the control sample, while probiotic counts ranged between 6.11 and 7.90 log CFU/mL in the samples produced with probiotic at the end of fermentation. They stated that the production by probiotic bacteria supported the functional properties of pickles.

The isolates used in pickle production were unable to grow at a pH 2. However, Hassanzadazar et al. [70] stated that the results of in vitro studies may not fully reflect the performance of probiotics in situ/foods, as many other physiological conditions may affect the survival of the strains. In in vitro analyses, all microbial cells are exposed to the same pH for the same length of time, whereas in food, not all cells are exposed to the same pH for the same length of time. The environment during static in vitro experiments can be much more inhibitory than the environment in actual food system (pickle), leading to the isolate’s inability to grow at pH 2 when cultured under in vitro experiments. Moreover, according to Ranadheera et al. [71], the selection of a suitable food matrix can potentially improve the acid tolerance of probiotic strains. On the other hand, the buffering capabilities of food ingredients may shield acid-sensitive bacteria from the acid [72]. Since the source of these isolates was pickles, the isolates may have adapted to the environment during fermentation and developed acid resistance, but not reflected in the in vitro presumptive probiotic count analysis.

After the fermentation process, microbiological counts were carried out regularly during storage (Fig. 2). The presumptive probiotic counts in the samples ranged between 1.38 log CFU/g and 6.51 log CFU/g during the storage. The counts decreased from the initial level of > 7 logarithmic units to 5–6 logarithmic units in the first week of the storage. The presumptive probiotic counts in all samples declined over the following weeks, depending on the probiotic type and time. The viable counts of PS1 and PS2 were found to decline significantly after 4 weeks, with these two samples exhibiting the lowest viable cell counts after storage compared to other samples enriched with probiotics (P < 0.05). Furthermore, PS3 (3.87 log CFU/g), PS4 (3.84 log CFU/g), and PS5 (3.62 log CFU/g) were not statistically different at the end of storage (P > 0.05). These samples had a higher count than NC. In PC, probiotic counts were lower than PS3, PS4, and PS5 during the storage (except for PS4 at the 4th week) (P > 0.05). Although the presumptive probiotic counts in the samples were at a level of 7 log CFU/g during fermentation (P > 0.05), they varied from 5.73 to 6.51 CFU/g at the beginning of storage and from 1.38 to 3.87 CFU/g at the end of storage (Fig. 2).

Nevertheless, these findings demonstrate that pickle samples containing high numbers of live probiotic cells can be produced by using pickle-derived cultures. Moreover, P. parvulus had the highest viability in pickles at the end of fermentation and storage, which can be associated with better growth of P. parvulus at low pH values and high salt concentrations [62]. Numerous studies have demonstrated that the viability and stability of probiotics in fruit and vegetable products differ significantly depending on the strain [68, 69]. In a study, cabbage juice was fermented using L. casei A4, L. plantarum C3 and L. delbruekii D7 and stored at 4 °C for 4 weeks. They stated that the probiotic count was 109 CFU/mL at the end of fermentation, but it decreased at the end of storage. They also stated that L. plantarum C3 and L. delbrueckii D7 counts were 4.1 × 107 CFU/mL and 4.5 × 105 CFU/mL, respectively, and that L. casei lost its viability at the end of storage [63].

A significant number of researchers have identified a decline in probiotic viability with storage, a finding that is widely acknowledged. Consequently, it is essential to use appropriate techniques, such as the addition of prebiotics, to produce probiotic foods.

Yeast counts in cucumber pickles during fermentation and storage

At the beginning of fermentation, the yeast counts were between 0.30 and 1.50 log CFU/g in the samples enriched with probiotics, while it was 1.04 log CFU/g in NC. During the following days of fermentation, the yeast counts increased, and the highest count was detected in PS2 with 4.34 log CFU/g on the seventh day of fermentation, and this sample was statistically different from PS3 and PS4 (P < 0.05). After the 7th day of fermentation, yeast counts decreased slowly in PS1, PS2, PS4, and PS5, while they increased in other samples (PS3, PC and NC). At the end of fermentation, the lowest yeast count among the samples enriched with presumptive probiotics was detected in PS4 (2.75 log CFU/g), which was statistically different from other samples (except PS2 and PS5) (P < 0.05). Furthermore, the highest yeast count was detected in NC (4.45 log CFU/g) (P < 0.05). NC had higher yeast counts than other samples (Fig. 3). This was because the probiotic culture was not added to compete with the natural microbiota in NC, and the antimicrobial metabolites produced by LAB were not present in the environment. The findings demonstrated that the fermentation process, when probiotics were introduced, effectively inhibited yeast growth. This is a beneficial outcome, as yeast typically results in an unfavourable alteration in the quality of the pickle. Consequently, the incorporation of probiotics offers a distinct advantage over the product obtained through spontaneous fermentation. Besides, the physical and sensory quality of foods is directly affected not only by yeasts but also by the presence of pectinolytic enzymes and volatile chemicals in the fermentation media [17].

Fig. 3

The radar chart illustrates the sensory properties of cucumber pickles at the end of fermentation and during storage

Following fermentation, the samples were monitored at regular intervals during the storage. At the end of fermentation, the yeast counts of the probiotic enriched samples showed a slight decrease from the beginning to the third week (Fig. 4). At the end of storage, the sample with the lowest yeast count belonged to PS1 with 2.42 log CFU/g (P > 0.05). Also, NC had a higher yeast count (3.02 log CFU/g) at the end of storage (P > 0.05, except PS1 and PS4). However, the yeast count of NC did not change statistically during storage (P > 0.05). In a similar study conducted by Turgut [64], cucumber pickles were produced by both spontaneous fermentation and using starter culture (L. plantarum). In their study, the highest yeast count was found on the 12th day of fermentation in the samples at 4.75 log CFU/mL, while in the control samples, the highest yeast count was determined at 5.56 log CFU/mL on the 11th day of fermentation (P < 0.05). These results were higher than the yeast counts that we had in our study. In the study performed by Alan and Yildiz [65], starter cultures (L. plantarum 8, L. plantarum 46, L. plantarum 61, L. paraplantarum and L. pentosus) were used in sauerkraut production as well as spontaneous fermentation. They found that the yeast count was 3.5 log CFU/g, but then the yeast count increased slightly (> 4.5 log CFU/g) until the end of the 15th day. The yeast count decreased after the thirtieth day and at the end of storage (an average of 5 log CFU/g). It was concluded that the use of Lactobacillus as a starter culture helped to decrease the total yeast-mold count in the samples. It is obvious that the results of this study are like our study.

In our study results showed that the probiotic counts continued to increase during the fermentation, with the highest levels reached during this period. Similarly, the probiotic counts decreased during storage, depending on time, food matrix and culture type. The probiotic count also remained constant at a certain logarithmic level at the end of storage, and these results were generally in line with the literature.

Sensory evaluation of the pickle samples

The samples were subjected to a sensory analysis at the end of fermentation period and with a 1-week storage interval for 5 weeks. The sensory characteristics of all samples were illustrated in Fig. 5. At the end of fermentation, PS1 had a higher color score (7.30) (P > 0.05, except PS5, PC and NC). In terms of odor characteristics, the most liked sample was PS1 (7.45) (P > 0.05), while PS5, PC and NC had lower odor scores (7.05) (P > 0.05). However, PS1 had a higher appearance score (P > 0.05, except PS5, PC and NC). The most liked sample in terms of taste was PS1 (7.25) (P < 0.05), while the lowest score (6.45) was found in NC. The results showed that the use of probiotics has a positive effect on the sensory properties of pickles. Endogenous cultures are thought to be more beneficial in this respect.

A review of the literature also showed that the use of endogenous cultures isolated from pickles and used as starter cultures in the production process is more beneficial than the use of industrial starter cultures isolated from different sources since endogenous cultures can adapt better to the environment and easily become predominant. Furthermore, endogenous starter cultures allow the production of safe and standard quality food [30, 66]. Hardness or crispness in pickle products is a valuable property sought by consumers, and it is important to preserve it during storage [67]. In this study, the highest crispness score was found in the PS1 (7.60) (P > 0.05), and it was significantly different from NC (P < 0.05). The addition of probiotics can achieve the desired level of crispness in pickles compared to spontaneous fermentation. Lastly, PS1 had a higher overall acceptance score (7.25) (P > 0.05, except for NC). As a result, the overall acceptance results revealed that pickle samples enriched with probiotics were generally more acceptable than control sample (NC).

Sensory characteristics of pickles were determined on a weekly basis through storage, and radar charts were used to illustrate the results, which were presented in Fig. 5. The objective was to demonstrate to the consumer whether there is a discernible difference in sensory characteristics when the products lose their probiotic properties and to facilitate a comparison of the products in terms of their sensory attributes. Nevertheless, even in the absence of an adequate number of probiotic cells in the product, the beneficial metabolites produced by these cells remain present [73]. Consequently, consumers may continue to derive benefit from these metabolites until the product is deemed acceptable.

During storage, color scores of the samples enriched with presumptive probiotics changed between 5.10 and 7.75, while it was between 4.65 and 6.50 for NC. The most liked sample on the 0th day of the storage was TS1 (P > 0.05, except PS5, PC and NC). At the end of storage, while the PS3 was the most liked sample (7.70) in terms of color properties (P < 0.05), the least liked sample was NC (P < 0.05). According to the results, all samples had acceptable scores in terms of color properties.

While the odor scores of the samples enriched with presumptive probiotics varied between 5.90 and 7.90, these scores were between 5.50 and 7.05 for NC. At the beginning of storage, PS1 was the most liked sample (7.45) (P > 0.05). At the end of storage, PS3 had the highest odor score (P < 0.05), while NC had the lowest odor score (P < 0.05). When the samples enriched with presumptive probiotics were compared for odor properties at the end of storage, it could be concluded that there was no significant difference between the samples except PS3 (P > 0.05), indicating that majority of the pickle samples underwent limited proteolysis that favorably contributes towards pleasant odor development instead of going through extensive proteolysis that would otherwise produces undesirable odor. Consequently, all samples were acceptable in terms of odor properties. The use of probiotics positively affected the odor characteristics of the samples.

The appearance scores of the samples enriched with presumptive probiotics varied between 5.25 and 8.80. These scores ranged from 4.60 to 6.25 in NC. PS1 had a higher score at the beginning of storage (P > 0.05, except for PS5, PC, and NC). At the end of storage, the sample with the lowest score (4.60) was NC (P < 0.05). The results revealed that the samples were acceptable in terms of appearance, and the samples enriched with presumptive probiotics were also more appreciated than the control sample.

The taste scores of the samples enriched with presumptive probiotics varied between 5.25 and 7.85, while these scores varied between 4.90 and 6.45 in NC during storage. PS1 was the most liked sample (7.25) at the beginning of storage (P < 0.05). At the end of storage, while the most liked sample (7.25) was PS3 (P < 0.05), the sample with the lowest score was NC (4.90) (P < 0.05) and the least liked sample was PC (5.25) (P < 0.05). These results showed that the samples were acceptable in terms of taste and using probiotics positively affects the taste of pickles.

While the crispness scores of the samples containing presumptive probiotics varied between 5.90 and 8.10, these scores varied between 5.80 and 6.90 in NC. At the beginning of storage, PS1 had the highest crispness score (7.60) and differed from NC (P < 0.05). In the last week of the storage, PS1 and PS3 had the same highest score (7.25). These samples were differed from the other samples except PS2 and PS4 (P < 0.05). Control samples, both PC and NC, had the lowest score (5.90) (P < 0.05). The results revealed that the samples were acceptable in terms of crispness. Hardness and crispness in pickles are important features search for consumers, and it is important to preserve them during storage (İcil, 2019).

The overall acceptance scores of the samples enriched with presumptive probiotics changed between 5.60 and 7.65 during storage, and these scores varied between 5.35 and 6.40 in NC. At the beginning of storage, PS1 had a higher overall acceptance score (7.25) (P > 0.05, except NC). In the last week of storage, PS3 had the highest overall acceptance score (6.70), and this sample was statistically different from the other samples (P < 0.05, except PS4). However, at the end of storage, the least liked sample was NC (P < 0.05) with the lowest score of 5.35. PC produced with the addition of L. plantarum 299v had the lowest score (5.70), which is statistically different from the other samples containing probiotics (P < 0.05, except PS5). The overall acceptance results revealed that all samples were generally acceptable. It was obvious that the overall acceptance scores were higher in the samples including probiotics, compared to NC during storage.