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Article

Development of Fermented Milks with Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 Isolated from Minas Artisanal Cheese

by
Camila Selles de Queiroz
1,
Bruna Samara dos Santos Rekowsky
1,2,
Madian Johel Galo Salgado
1,2,
Maria Fernanda Barreto da Hora Lopes
1,2,
Marcelo Resende de Souza
3,
Marion Pereira da Costa
1,2 and
José Givanildo da Silva
1,2,*
1
Laboratory of Technology and Inspection of Milk and Derivatives (LaITLácteos), School of Veterinary Medicine and Animal Science, Federal University of Bahia (UFBA), Ondina, Salvador 40170-110, BA, Brazil
2
Program in Animal Science in the Tropics (PPGCAT), School of Veterinary Medicine and Animal Science, Federal University of Bahia (UFBA), Ondina, Salvador 40170-110, BA, Brazil
3
School of Veterinary Medicine, Federal University of Minas Gerais (UFMG), São Francisco, Belo Horizonte 31270-901, MG, Brazil
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(10), 560; https://doi.org/10.3390/fermentation11100560 (registering DOI)
Submission received: 27 August 2025 / Revised: 15 September 2025 / Accepted: 23 September 2025 / Published: 28 September 2025
(This article belongs to the Special Issue Traditional and Innovative Fermented Dairy Products)
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Abstract

The aim of this study was to develop fermented milks using strains of lactic acid bacteria with probiotic potential isolated from Minas artisanal cheese. For this purpose, the strains Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 were used in a 6 × 4 experimental design, with six fermented milk treatments and four analyses on days 1, 15, 30 and 45 to characterize the product and evaluate the viability of the bacteria. Additionally, a sensory analysis was conducted using a preference ranking test. All treatments showed viable counts of microorganisms above 106 CFU/mL until the end of the storage period, as well as variations in pH and titratable acidity values on day 45, being the lowest value of both verified in the TRAT1B7 sample. Based on the application of a sensory analysis, it was observed that the type of treatment and the strain directly impacted the sample chosen by the tasters. According to results gathered from the present study, milk fermented by L. casei B5 stands out in the order of preference, however milk fermented by L. plantarum B7 has also obtained favorable results, indicating that this strain of bacteria can be used in dairy technology.

1. Introduction

The global trend towards improving individual health and the growing public awareness in search of more beneficial foods has been driving the food industry to enhance its range of so-called functional products. In this context, fermented milk has gained prominence in the market, as consumers seek foods with pleasant flavor and aroma, low caloric content and positive health effects [1]. Certain fermented milks deliver health benefits stemming from the inclusion of Lactic Acid Bacteria (LAB) with probiotic characteristics, such as certain strains of Lactobacillus spp., including Lacticaseibacillus casei and Lactiplantibacillus plantarum [2].
Lactobacillus spp. are among the microbial genera that constitute Lactic Acid Bacteria (LAB), widely used in the production of dairy products, including fermented milk. In these products, lactic acid-producing bacteria are used for fermentation, as their addition can extend the shelf life by lowering the pH, while also producing various compounds that contribute to functional activities, such as biopreservation [2]. Moreover, the intake of these foods can offer health benefits to consumers [3].
These microorganisms can support the balance of the intestinal microbiota, thus benefiting the host [4]. It has broad technological applications and is considered highly safe for use in food formulations, as these bacteria are non-pathogenic and do not transmit resistance genes to other microorganisms [5].
Due to the technological properties and health benefits previously, the research has focused on identifying and characterizing new strains of Lactobacillus spp. to develop innoval dairy products. In this context, artisanal products are seen as potential sources of such strains. Among Brazilian artisanal products, there is the Minas cheese, which is traditionally produced using raw milk, endogenous starter cultures (named pingo), rennet and salt. Although this preserves traditional cultural heritage, it restricts commercial distribution due to sanitary regulations [6]. Nonetheless, the way of cheesemaking contributes to cultural richness, added value, and product differentiation [7]. During artisanal cheese ripening, microorganisms grow and promote biochemical alterations, with LAB among them so named because they ferment carbohydrates and produce lactic acid. Among these LAB, Lactobacillus spp. is one of the most frequently desirable microorganisms found in cheeses [3].
Therefore, the objective of this study was to develop and characterize fermented milk products using strains of L. casei B5 and L. plantarum B7 with probiotic potential, previously isolated from Minas artisanal cheese. Moreover, their viability was assessed under different manufacturing methods/scenarios, as well as the sensory analysis to determine Purchase intent.

2. Materials and Methods

2.1. Raw Material Acquisition

For the preparation of the fermented milk samples, skimmed milk powder, UHT skimmed milk, and sucrose were purchased from a retail market in the city of Salvador, Bahia, Brazil, ensuring the same brand and batch number for each item.
Skimmed milk powder (Amount per 20 g serving): Total fat 0.0 g; basic ingredients: skimmed milk. UHT skimmed milk (Amount per 100 mL serving): Total fat 0.0 g; basic ingredients: skimmed milk, stabilizers pentasodium triphosphate, trisodium citrate, sodium dihydrogen phosphate, and disodium diphosphate.
Quality control of the raw materials (skimmed milk powder and UHT milk) was carried out in accordance with the Technical Regulation of Identity and Quality (RTIQ) established by Brazilian legislation [8,9]. For microbiological control of skimmed milk powder, analyses were performed on the global count of aerobic mesophilic microorganisms (CFU/mL), count of enterobacteria (CFU/mL), count of Staphylococcus spp. and research of Salmonella spp.; For physicochemical control, titratable acidity and fat percentage were determined. For the microbiological and physicochemical control of UHT skimmed milk, a global count of mesophilic aerobic microorganisms and determination of titratable acidity and fat content were performed [8,9,10]. All analyses were performed in triplicate.
The strains L. casei B5 and L. plantarum B7 used as starter cultures for the development of the fermented milk products were previously isolated and identified from artisanal Minas cheese, and their probiotic properties had been assessed in vitro [11]. These strains were kindly provided by the Department of Technology and Inspection of Animal Origin Products at the School of Veterinary Medicine at the Federal University of Minas Gerais (UFMG).

2.2. Activation and Fermentation Curve of LAB Strains

The strains were frozen at −20 °C and were activated by transferring 1 mL of each LAB into test tubes containing 9 mL of MRS broth (Man Rogosa & Sharpe—Merck), followed by incubation at 37 ± 2 °C for 24 h. After then growth curves were performed.
Six aliquots of each strain were made, of which 2 aliquots were determined for each study time, being T0, T1 and T2. For both, T0, serial dilution in 0.1% peptone water (Merck, Oxoide Limited, Basingstoke, UK) was performed (from 10−1 to 10−8) and plated on MRS Agar (Man Rogosa & Sharpe—Merck, Oxoide Limited, Basingstoke, UK), in duplicate, and the plates were incubated for subsequent colony counting. For L. casei, T1 corresponded to 5 h after T0 and T2, 10 h after T0. For L. plantarum, T1 was 9 h after T0 and T2, 18 h after T0. The same serial dilution and plating protocol was followed for all time points. After 48 h of incubation, all plates were examined, and colony-forming units (CFUs) were counted.
The purpose of the curve was to observe the growth pattern of the bacteria, determining in which period the strains reached the minimum concentration of 106 CFU/mL. There was a difference in incubation temperature for the two strains; in the case of L. casei, a temperature of 37 °C ± 2 °C [12] was used, and for L. plantarum, 30 °C ± 2 °C [4].
Following this, the fermentation profile was determined by measuring pH, titratable acidity, and counts of lactic acid-producing bacteria (CFU/mL) on MRS Agar (Man Rogosa & Sharpe—Merck). These analyses were performed at each 2-h time intervals from time zero (addition of the inoculum, 3% of the culture at 106 CFU/mL in 200 mL of skim milk) until the coagulation of the milk added with the strains of L. casei and L. plantarum. Fermentation time was also determined during the curve, with the initial time point defined as inoculum addition and the final point as coagulum formation [13].

2.3. Development of Fermented Milk Samples

A 6 × 4 factorial design was used, involving six fermentation treatments (three for each bacterial strain) and four evaluation periods of storage: days 1, 15, 30, and 45. Each treatment and strain were assigned specific codes, with “B5” referring to L. casei and “B7” to L. plantarum (Chart 1).
Treatment 1 consisted of skimmed milk powder reconstituted at 10% in distilled water, added with 8% sucrose, followed by heat treatment in an autoclave at 110 °C/10 min. After heat then, 3% of LAB culture was inoculated. The codes assigned to this treatment were TRAT1B5 and TRAT1B7.
For the second treatment, skim UHT milk added 8% sucrose was inoculated with 3% of LAB and products were designated as TRAT2B5 and TRAT2B7
Treatment 3 used the same ingredients as Treatment 1, but with prior sterilization of the distilled water before reconstituting 10% milk powder and 8% sucrose, 3% of LAB culture was inoculated. This last treatment received the codes TRAT3B5 and TRAT3B7.
In all treatments, LAB cultures were inoculated when the mixtures reached 37 °C.
After inoculation, all fermented milk samples were incubated at 37 °C until the formation of a coagulum, which marked the end of fermentation. The appearance of the coagulum corresponded to the isoelectric point of casein, approximately pH 4.6. After coagulation, the products were stored at 8–10 °C for 45 days. Physicochemical and microbiological analyses were conducted on days 1, 15, 30 and 45 of storage [13].

2.4. Physicochemical Analyses

For the pH analysis, an aliquot of approximately 20 mL from each sample was measured using a benchtop/portable pH meter (model LUCA210®, Lucadema, São José do Rio Preto, São Paulo, Brazil).
Titratable acidity was determined following the methodology established by IDF Standard 150:1991 [14], in accordance with the Official Methods Manual for the Analysis of Products of Animal Origin from the Brazilian Ministry of Agriculture and Livestock (MAPA) [15]. All analyses were performed in triplicate.

2.5. Microbiological Analyses

Microbiological analyses were conducted in accordance with the Technical Regulation of Identity and Quality (RTIQ) for fermented milk products [16].
Counts of lactic acid bacteria were performed from serial dilutions of the fermented milks in 0.1% peptone water. Subsequently, 0.1 mL of each dilution was inoculated onto MRS agar plates. The plates were incubated at 37 °C for 48 h under microaerophilic conditions. After the incubation period, colony counts were performed, and results were expressed as colony-forming units (CFU), according to the methodology described in IDF Standard 117:1988 [17].
For total and thermotolerant coliform counts, the Most Probable Number (MPN) method was followed IDF Standard, 73:1985 [15,18].
Molds and yeasts detection was also performed. For this, 0.1 mL of each dilution was inoculated in duplicate onto Petri dishes containing DRBC agar (Dicloran Rose Bengal Chloramphenicol—Merck) and incubated at 25 °C for 5 to 7 days APHA 21:2015 [19].

2.6. Sensory Analysis

All evaluators provided voluntary informed consent prior to participation. This study was approved by the Ethics Committee on Research in Human Beings of the Federal University of Bahia, Brazil (National Health Council, Resolution No. 196/1996) approval by the Ethics Committee of School of Nursing of Federal University of Bahia (CAAE 60414022.7.0000.5531; approved on 1 March 2023).
The sensory analysis was conducted using a preference ranking test, as described by another study [20]. The test was carried out with volunteers from the School of Veterinary Medicine and Animal Science at the Federal University of Bahia. Before starting the analyses, the tester was briefed on how the test would work, how to fill out the form and was asked to read, fill out and sign the Free and Informed Consent Form (TCLE), thus authorizing the start of the analyses.
A total of 75 untrained consumers voluntarily participated in the sensory analysis. None of the participants reported lactose intolerance or milk allergies.
In addition to the consent form, each participant received an evaluation form and six coded samples (identified by a sequence of three random digits), in refrigerated portions of 15 mL (8 °C ± 2 °C), in disposable plastic cups. The evaluation form included a section for personal information and instructions to rank the six samples in crescent order of preference, from most to least preferred sample. The aim was to determine which sample had the highest acceptance and whether the treatments and/or bacterial strain influenced the order of preference. An optional space for comments was also provided.

2.7. Statistical Analysis

The results obtained from pH and titratable acidity measurements were statistically analyzed by Analysis of Variance (ANOVA), with Tukey test used for mean comparisons. A 5% significance level was adopted.
For the LAB counts, the means were also evaluated through Analysis of Variance (ANOVA), and the Tukey test was used to compare them, with a significance level of 5%.
Results from the preference ranking test were evaluated using the Friedman test to determine whether there were statistically significant differences among samples (p < 0.05) [14]. Scores were assigned based on the ranking position (1 to 6), in which the most preferred sample had the lowest total score and the least preferred had the highest.
The ordering was assessed using the Friedman test, with the help of the Newell and McFarlane Table, which provides the critical difference for an ordering test with 75 participants and 6 samples, equal to 64, at a significance level of 5% [14].

3. Results

Quality control analyses were performed on the skimmed milk powder and UHT milk used as raw materials. In the microbiological analyses performed, no growth of aerobic mesophilic microorganisms was observed (in the skimmed milk powder and UHT samples); enterobacteria, Salmonella spp. and Staphylococcus spp. For skimmed milk powder, the average fat value was 0%, while the acidity value was 1.2% (g lactic acid/100 g). In skimmed UHT milk, an average fat value of 0% and titratable acidity of 0.14 g of lactic acid/100 g were also found. The results indicated compliance with current Brazilian legislation and Codex Alimentarius Standard and confirmed their suitability for consumption and production of fermented milks [10,15].
According to the growth curve, under the following time and temperature conditions, L. casei B5 at 37 °C/48 h and L. plantarum B7 at 30 °C/48 h, obtained counts of 5.55 × 107 CFU/mL and 2.2 × 107 CFU/mL, respectively. In the fermentation profile, coagulation occurred after 24 h of incubation.
All treatments showed a reduction in pH throughout the analyses, from day 1 to day 45 (Table 1). We could observe that on day 1, the sample with the highest pH value was TRAT1B5, with 4.15, and the lowest value was TRAT1B7, with 3.76. However, TRAT1B5 presented greater variation in pH values throughout the storage period than TRAT1B7. The results of the average pH and titratable acidity values can be seen in Table 1 and Table 2, respectively.
The treatments TRAT1B5, TRAT2B5, and TRAT3B5 showed an increase in titratable acidity values from day 1 to day 30; however, a decrease was observed on day 45. Conversely, the samples TRAT1B7, TRAT2B7, and TRAT3B7 exhibited a different behavior over the storage period (Table 2).
The means of lactic acid bacteria (LAB) counts for each treatment during the storage period are presented in Table 3.
On day 1, the treatments showed similar LAB counts (Figure 1), ranging from 8.22 to 8.49 Log CFU/mL with minor variations (p > 0.05), except for TRAT2B7, which exhibited significantly lower growth (p < 0.05). This indicates that L. plantarum B7 had a slower fermentation time compared to L. casei B5 in UHT milk; however, despite the variations, initial bacterial viability appeared similar.
On day 15 (Figure 1), variation in counts among strains and their respective treatments began to be observed. On day 30, a pronounced difference was noted in TRAT2B7. By day 45, a reduction in bacterial counts was observed across all treatments and strains used (Figure 1).
Quality control analyses of the fermented milks showed that the counts of total coliforms were <3 MPN/g or mL. No growth of molds or yeasts was observed.
The results obtained from the preference ranking test are presented in Table 4. Based on these results, the sample T1LC had the lowest total score, indicating it was the most preferred. The table also shows the total scores for each treatment.

4. Discussion

The results obtained from the growth curve indicate a LAB count of 107 CFU/mL was reached within 48 h, being sufficient to adapt both strains to Brazilian legislation, which requires a minimum concentration of 106 UFC/mL, from the preparation until the end of the storage period of fermented milk [16].
Different plating times and temperatures were applied from the growth curve for each strain according to their characteristics. The differences between incubation times and temperatures were based on the literature consulted. L. casei was incubated at 37 °C, its optimal growth temperature [12], while L. plantarum, being a mesophilic organism, showed better development from 30 to 35 °C [4], with a lag phase of 3 to 4 h, an exponential growth phase lasting 18 h, followed by a stationary phase lasting 4 h [21].
The results of the quality control analyses of skimmed milk powder and UHT confirmed their suitability for the production and consumption of fermented milks. The results indicated compliance with current Brazilian legislation and Codex Alimentarius Standard [10]. The objective of verifying the microbiological and physicochemical quality of skimmed milk powder and skimmed UHT milk was to characterize the milks used and, consequently, ensure quality control of the production of fermented milks.
All fermented milks showed LAB counts above the minimum established by the Brazilian Technical Regulation for Identity and Quality of Fermented Milks (RTIQ) throughout the 45-day shelf life, with probiotic-appropriate levels. These results demonstrate that the established time period can be adopted as the shelf life for these products.
Although both strains (L. casei B5 and L. plantarum B7) belong to species characterized by facultative heterofermentative metabolism, L. casei B5 showed superior adaptive ability from day 1, as observed in Table 3. This behavior was observed from the initial curve development. This metabolic characteristic of fermentation allows the microorganism to adapt to the environment and substrate, enabling its development. In Table 3, we can observe that there was a significant difference between the treatments, especially on days 30 and 45, when they presented divergent groups when comparing them, so we can raise the hypothesis that when varying the type of treatment applied, there were different responses, as well as different metabolic characteristics between the strains used [22].
When analyzing the pH and acidity results, comparing the different fermented milk treatments and the days, as well as the difference between the two strains used in the experiment, we can observe that the pH values (Table 1) presented statistical differences over the days, regardless of the strain and treatment applied. Titratable acidity (Table 2) demonstrated statistical differences for both strains and treatments. It is worth noting that the titratable acidity values observed in all treatments during storage are in accordance with the standards established by Brazilian legislation [16].
However, no pH standards are established for fermented milks by current legislation, making it impossible to compare the values found with current legislation [16] nonetheless the values presented here (Table 1) align with those found previous studies [1], who used another L. plantarum strain isolated from Minas artisanal cheese, reporting a mean pH of 4.1. Furthermore, the low pH observed in all treatments contributes to the development of acidity, a desirable sensory characteristic in this kind of product. We also observed that the pH value was not far from that reported in commercial fermented milks, which range from 3.61 to 4.11 [23].
The reduction in pH did not impair probiotic microorganism viability, as LAB can stay viable at pH levels as low as 3.8 while still producing enzymes [5]. Treatment TRAT1B7 exhibited the lowest pH on day 45 (Table 1), a result expected in fermented products by LAB. The literature [22] points out that with the breakdown of lactose, exocellular metabolites, such as lactic acid, help to increase the shelf life of foods and contribute to their biopreservation [2].
As shown in Table 1, on the first day of storage, pH values varied from 3.76 to 4.15. On the last day (45 days), even under refrigeration, a reduction was observed, with average pH values ranging from 3.55 to 3.83 in the different treatments. We can infer that this variation is a result of the metabolic activity of the LABs used [5]. TRAT1B5 and TRAT2B5 showed progressive pH reduction throughout all storage days. In contrast, TRAT3B5 exhibited pH decreases only on days 1, 15, and 30, with no significant difference from days 30 to 45.
TRAT1B7 showed no difference in pH values on days 1 and 15, but thereafter, there was a reduction in pH. TRAT2B7 showed a significant reduction in pH during the storage period. On the other hand, TRAT3B7, showed a reduction in pH values until day 30. On day 45, there was an increase in pH value, contrary to expectations, which would be a continuous reduction in pH over time. This behavior was consistently observed across all treatments, as microbial growth led to sustained lactose consumption and consequent lactic acid production throughout the fermentation process. According to another author [24], throughout the fermentation process, lactic acid bacteria can trigger proteolysis in the food, where pH changes (increases) can occur due to the release of ammonia during proteolytic reactions.
The titratable acidity of the treatments on day 1 of storage ranged from 1.17 g of lactic acid/100 g to 1.60 g of lactic acid/100 g, as shown in Table 2. During storage, acidity ranged from 1.11 g lactic acid/100 g to 1.78 g lactic acid/100 g. The lowest titratable acidity value was on day 15, verified in TRAT3B7, of 1.07 g of lactic acid/100 g, and the highest value on day 30, in TRAT2B5, of 1.81 g of lactic acid/100 g. It can observe, from this data, that both strain and type of heat treatment directly influenced the variation in the average titratable acidity. The lowest value was presented by a treatment (TRAT3B7) that used L. plantarum and the highest value was presented by the one that used L. casei, (TRAT2B5), confirming the difference in metabolism, as previously mentioned, and the adaptation of each strain [24]. Also, according to the results presented in Table 2, the treatment that presented the smallest variation and lowest acidity values was TRAT3B7, suggesting better adaptation of the strain used.
In 2023, a study showed [25] stated that acidification even after cooling samples can be explained by the remaining metabolic activity of the cultures. Similarly, variations between strain values can be attributed to the improvement efficiency of each strain [22]. According to the results of this study, L. plantarum was the most efficient strain in the fermentation process, as TRAT3B7 demonstrated the lowest variation in titratable acid and pH, as well as exhibiting the highest average LAB count on day 45 (8.47 log CFU/mL).
A decrease in pH values over storage time was expected, and an inversely proportional increase in acidity [26,27]. However, in this study, we observed that pH had a general tendency to decrease, but acidity values were not considered in the same proportion. It is believed that these characteristics can be explained by the method used to determine this acidity (titratable acidity). When we examine the variations in titratable acidity between the samples, we can see that, as shown in Table 2, TRAT1B5 increased from day 1 to day 30, and on day 45 there was a decrease in this acidity, in contrast to the pH values, which maintained a decrease throughout the period.
The pH values tend to decrease as titratable acidity values increase, however, the methodology for determining titratable acidity used in this study is specific for measuring lactic acid per 100 g of sample [14]. Still, the pH measurement methodology evaluates the acidity of the product as a whole, this could explain the different results of the analyses [23]. Similarly, the pH reduction is evidenced by the growth of the strains and the increase in their count (CFU/mL), as shown in Table 1 and Table 3. According to their metabolism, LABs use the conversion of lactose to lactic acid during fermentation, but the pH methodology is not specific to this. Analyzing this treatment (TRAT1B5) specifically, it can be inferred that the change in values from day 1 to day 15 is due to the consumption of this lactose, which, from day 15 to day 30, would already be decreasing, resulting in a reduction in values until the last day of analysis [23,27].
TRAT2B5, at the first three measurement points, showed an increase in titratable acidity values in agreement with the decrease in pH. However, from day 30 to day 45, the reduction in acidity value was slight, possibly indicating a decrease in metabolic activity. This fact is also reflected in the pH values, in which there was no significant difference from days 30 and 45. It is worth noting that TRAT3B5 presented the same behavior (both pH and titratable acidity) as TRAT2B5.
TRAT1B7 showed a decrease in titratable acidity values from day 1 to day 15, with a slight increase from day 15 to day 30, and from day 30 to day 45. This slight increase may indicate the strain adaptation time to the culture medium, as did the pH values, which showed the same fluctuation. There was no difference in pH values on days 1 and 15. From day 15 to day 30, there was a significant reduction, but on day 30 and day 45 the reduction was slight. TRAT2B7 showed divergent behavior both in pH and titratable acidity values (see Table 1 and Table 2).
TRAT3B7 initially showed a decrease in titratable acidity (from day 1 to day 15), while at other points, there was a slight increase until day 45. The pH decreased on Days 1, 15, and 30, but this was the only sample in which the pH increased from day 30 to day 45, diverging from expectations. We can suggest an occurrence of proteolysis resulting from the enzymatic activities of the strains, as reported by [23,28], corroborating the report of “lumps” in the sensory analysis.
In Type 1 treatments (TRAT1B5 and TRAT1B7), in which all inputs were autoclaved together (see Table 1), there was a tendency for pH to decrease and acidity to increase over the analysis time. This trend is characteristic of fermented milk production, but it is necessary to control the pH to prevent the product from becoming too acidic and impacting the final product quality. However, the fact that all inputs were subjected to the autoclave process together may have directly affected the acidity of the final product, as this process triggered the Maillard reaction [12,29], which slightly acidifies the medium coupled with potential denaturation of the milk protein [28,29], which may have altered buffering capacity of the medium, in other words, its resistance to pH variations. This can make the medium more susceptible to pH drops.
Other authors have made an assessment [12] analyzed 24 strains of the Lactobacillus spp. in autoclaved medium and concluded that some strains, such as L.acidophilus, showed better growth and higher cell counts in autoclaved medium, possibly due to heat-generated Maillard reaction products.
According to some studies [30,31], skim milk powder is added before heat treatment to increase the defatted dry extract index and thus avoid the syneresis authors argue that [26] that L. casei has a positive impact on the quality parameters of dairy products, such as reducing syneresis resulting from the production of exopolysaccharides. These exopolysaccharides contribute to food texture, improving viscosity and consequently increasing firmness, thus making the food more attractive to consumers, demonstrating that this is a desirable characteristic [4]. The combination of powdered milk and L. casei may have acted synergistically to reduce syneresis and consequently contributed to improved sensory characteristics.
In type 2 treatments (TRAT2B5 and TRAT2B7), fermented milk made with UHT milk, there was an increase in acidity and a reduction in pH over the days of analysis, but with a tendency towards stability, as can be seen in Table 1 and Table 2. According to the Technical Regulation for the Identity and Quality (RTIQ) of fermented milks [16], these products must have an acidity from 0.6 to 2.0 g of lactic acid/100 g. Therefore, we can state that all fermented milks developed in this research comply with the legislation, which corroborates the desirable physical and chemical characteristics for the viability of probiotic microorganisms [32]. However, the acidity values in the present study differ from those of fermented milks marketed in Brazil; according to [1], their variation is from 0.88 to 1.17 g/100 g. This discrepancy between the data from this study and the literature suggests that the type of treatment applied to the samples may have influenced these values. Furthermore, the origin of the strains used in this experiment demonstrates that they were better adapted to the dairy matrix, having been isolated from a dairy product.
In the LAB microbiological count, we observed that L. plantarum B7 exhibited satisfactory growth; however, when compared to L. casei B5, the latter was more demanding, exhibiting slower growth (Table 3). Both strains presented homogeneous and numerous counts at dilutions of 10−6, including the counts performed on the 45th day of analysis. This indicates that the LABs remained viable within the expiration date established in this study, thus complying with Brazilian legislation [16].
However, considering different studies [5] that reported that milk fermented with L. casei maintained its recommended population for three weeks, unlike the results obtained here, where they remained viable until the last day (equivalent to approximately 6 weeks) of storage. This fact indicates that the strains used in this study have different metabolic characteristics than those cited by [5], allowing greater viability. It is also important to emphasize that these strains originated from a dairy product and are better adapted to the dairy matrix, resulting in greater viability. Among the Lactobacillus group, the species L. casei is one of the most researched. Several studies have shown that this strain has a greater survival ability even after temperature variations. Some strains also have the ability to produce thermostable bacteriocins, creating the possibility of technological application as biopreservatives [22,30].
According to Table 3, we can observe the LAB growth between treatments, where on day 1 there was no statistically significant difference between them. However, on day 15, the TRAT1B5 treatment was equal to TRAT3B7, while the others were equals between themselves. On day 30, TRAT1B7 was different from all the others, while TRAT2B7 was equal to TRAT3B5, and the others formed another group with no statistical difference. On the last day of analysis, the TRAT1B7, TRAT2B7, and TRAT3B5 treatments differed from each other and from the others.
Also, according to Table 3, we can see the difference in results on each day of analysis. On the first day of storage, all fermented milks showed microbiological growth, with no statistical difference. After 15 days of storage, a variation between the counts was evident, with TRAT1B5 and TRAT1B7 differing from each other and from the other treatments. TRAT2B5, TRAT2B7, and TRAT3B5 were equal to TRAT3B7. On day 30, TRAT1B5 showed statistically significant differences in LAB counts compared to all treatments, as did TRAT1B7 and TRAT3B7. However, TRAT2B5, TRAT2B7, and TRAT3B5 were equal between themselves. On day 45, we observed that TRAT1B5 and TRAT1B7 were different from all other samples and from each other, while the others formed a homogeneous group.
The results of the coliforms, molds and yeast counts indicated no microbiological growth throughout the experiment, ensuring the quality of the fermented milks and their microbiological safety, as required by law [16]. It should be noted that the results of the coliform analysis and the mold and yeast count indicate good handling practices during food preparation, as these microorganisms are indicators of hygienic quality, as well as ensuring that no contamination occurred during the storage period [27]. According to MAPA [15], fermented milks should not contain thermotolerant coliform values above 10 MPN/mL, as well as the molds and yeast counts, which allows a maximum of 200 CFU/g [1].
From the data obtained in the ranking test, presented in Table 4, we can observe that TRAT1B5 was the preferred simple among the volunteers, followed by TRAT1B7 and TRAT3B5. Thus, the TRAT3B7 sample is statistically different from all samples, as well as being the least preferred, according to the preference ranking test. The TRAT2B7 sample differs from TRAT1B5, while TRAT2B5 differs from TRAT1B5. Although the TRAT1B5 sample was the preferred sample among the volunteers (smaller total, see Table 4), statistically there is no difference in relation to the samples TRAT3B5, TRAT1B7 and TRAT1B5.
It is clear that the treatments that used UHT milk were not the most preferred among the volunteers. Likewise, the treatments that included L. plantarum in their formulation were not among the most preferred groups, with the exception of the TRAT1B7 sample. In this case, we can suggest that the type of heat treatment applied to this sample possibly positively influenced its sensory perception, despite L. plantarum being part of its formulation and not being the strain that stood out in the sensory analysis. It is believed that the heat treatment may have improved the tasters’ perception of this sample due to the caramelization process (Maillard reaction) which compounds resulting from heating [29].
The two strains have similar metabolisms and may have the ability to produce flavoring compounds under certain circumstances, leading the food industry to add not only probiotics for their acidifying properties, but also aromatic substances, as stated by another studies [21,33]. It was found that L. casei B5 provided the best sensory characteristics for the test participants, as it was the most similar to the popular industrialized fermented milk, sold in Brazil, considered a reference among the volunteers. During the analysis of the evaluation sheets, it was found that the tasters cited the “characteristic flavor” of fermented milk, very similar to a well-known brand, and praised the consistency of the TRAT1B5 sample.
On the other hand, the TRAT3B7 sample was considered “unpleasant” due to its texture; some tasters noted the presence of lumps, which leads us to believe that it was not as homogeneous as the others [28] indicated that these processing defects may be associated with poor dissolution of the milk powder, albumin denaturation, and/or calcium phosphate precipitation. Texture changes due to the breakdown of protein molecules and increased binding with water molecules can lead to the formation of lumps, adding to the change in the texture of the final product. This could corroborate this hypothesis, as these “lumps” were observed in the TRAT1B7 sample reported in the sensory analysis. Findings have been reported where fermented milk supplemented with L. plantarum demonstrated greater syneresis compared to other products during refrigerated storage (approximately 30–36%) [34].
L. plantarum [33] is part of a group of LAB considered non-starter (NSLAB), and therefore, it could add unpleasant characteristics to dairy technology. The authors state that NSLAB are correlated with technological defects that impact product flavors and aromas, including textural changes, off-odors, and even possible bitter tastes [24].
L. plantarum is not commonly used in the production of fermented milks. However, based on the results obtained in this study, we can indicate that it has potential for use in the production of fermented milks, depending on the formulation used. The type of treatment influenced the choice of the preferred sample, according to the sensory analysis, with preference being given to those made from 10% reconstituted milk powder and autoclaved before adding the inoculum. These data corroborate the use of milk powder in the production of fermented milks, as the increase in defatted dry extract increases the water retention capacity of the proteins, preventing the separation of water and curd, and increasing the consistency, texture, and viscosity of the final product [30].
The TRAT1B5 treatment exhibited homogeneous microbial growth, with no statistical difference over the storage time. It was the sample with superior sensory characteristics compared to the others and also presented low variation in titratable acidity values. Thus, it was the treatment that demonstrated technological potential combined with public acceptance.

5. Conclusions

This study confirms the feasibility of developing fermented milk beverages using L. casei B5 and L. plantarum B7, achieving satisfactory physicochemical profiles and microbiological stability over a 45-day storage period.
According to the sensory analysis, treatment type 1 was the most accepted, and the L. casei B5 strain also outperformed the other strains used. These variables contributed to the greater acceptance of TRAT1B5. However, TRAT1B7 fermented with L. plantarum B7 also stood out, being one of the most accepted. Considering this, it was observed that the type of treatment resulted in a greater preference than the type of strain (TRAT1B5 and TRAT1B7 were the preferred samples). This public response can be explained due to the Maillard Reaction which resulted in a more pleasant “flavor” for the tasters. Therefore, we can conclude that L. plantarum B7 can also be implemented in dairy food technology, as long as the treatment is adapted to maximize its positive sensory characteristics.
Despite the reduction in the concentration of CFU/mL throughout the period of this research (45 days), the results still demonstrated that even on the last day of analysis, concentrations above 106 CFU/mL were reached in all treatments, demonstrating that the shelf life can be extended, as long as it is kept under refrigeration (2 to 8 °C), ensuring its viability.
Based on the results obtained, it is possible to consider the potential for technological and industrial application of lactic acid bacteria of artisanal dairy origin for product development. The current study highlights the importance of using autochthonous LAB strains in fermented dairy products aid in the development of foods, as they contribute to the shelf life of dairy products and can help improve the sensory characteristics of products.

Author Contributions

Conceptualization, C.S.d.Q. and J.G.d.S.; methodology, C.S.d.Q., M.J.G.S. and J.G.d.S.; software, C.S.d.Q. and B.S.d.S.R.; validation, C.S.d.Q. and M.P.d.C.; formal analysis, C.S.d.Q.; investigation, C.S.d.Q., B.S.d.S.R., M.J.G.S., M.R.d.S., M.F.B.d.H.L., M.P.d.C. and J.G.d.S.; resources, C.S.d.Q. and J.G.d.S.; data curation, C.S.d.Q.; writing—original draft preparation, C.S.d.Q.; writing—review and editing, C.S.d.Q.; visualization, C.S.d.Q. and M.F.B.d.H.L.; supervision, J.G.d.S. and M.P.d.C.; project administration, C.S.d.Q.; funding acquisition, C.S.d.Q. and J.G.d.S. All authors have read and agreed to the published version of the manuscript.

Funding

The authors gratefully acknowledge the financial support provided by for granting research grants to the first author and to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the financial support (Process number 404304/2022-2).

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Review Board (or Ethics Committee) of Human Research Ethics Committee of the Federal University of Bahia (process number 60414022.7.0000.5531) for studies involving humans.

Informed Consent Statement

Informed consent was obtained from all subjects involved in the sensory analysis, and the study received ethical approval from the Human Research Ethics Committee of the Federal University of Bahia (process number 60414022.7.0000.5531).

Data Availability Statement

The datasets generated for this study are available on request to the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest and that the funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

References

  1. Gusmão, A.C.M.; Neves, L.F.; Matos Junior, F.E.; Cardoso, T.S.; Menegali, I.; Pereira, S.; Costa, T.O.; Pinto, M.S. Probiotic Potential of Fermented Milk Containing Lactobacillus rhamnosus and Lactobacillus plantarum Isolated from Artisanal Cheeses from Northern Minas Gerais. Rev. Inst. Laticínios Cândido Tostes 2023, 78, 924. [Google Scholar] [CrossRef]
  2. Cancella, M.J.; Santos, T.A.; Miguel, E.M.; Moreira, G.D.M.M.; Sobral, D.; de Almeida, F.A. Lactic Acid Bacteria as Probiotics, Biopreservatives, and Bacteriocin Producers: A Review. Rev. Inst. Laticínios Cândido Tostes 2024, 79, 37–47. [Google Scholar] [CrossRef]
  3. Kegele, C.S.; Valladares, L.S.; Silva, J.F.M.; Alonso, B.V.; Ribeiro, J.B. Technological Potential of Lactic Acid Bacteria as Biopreservants: Literature Review. 2023. Available online: https://agronfoodacademy.com/potencial-tecnologico-de-bacterias-acido-laticas-como-bioconservantes-revisao-de-literatura/ (accessed on 22 September 2025).
  4. Freire, T.T.; Silva, A.L.T.; Ferreira, B.K.O.; Santos, T.M. Lactic Acid Bacteria: Characteristics and Importance—Review. Res. Soc. Dev. 2021, 10, 19964. [Google Scholar] [CrossRef]
  5. Mello, T.C.S. In Vitro Probiotic Properties of Lactic Acid Bacteria Isolated from Artisanal Minas Cheese from Campo das Vertentes, Minas Gerais. Master’s Thesis, Federal University of Minas Gerais, Belo Horizonte, Brazil, 2022. [Google Scholar]
  6. Brazil. Law No. 13,860 of 18 July 2019. Provides for the Production and Commercialization of Artisanal Cheeses and Other Provisions. Off. Gaz. Union 2019, 138, 1. [Google Scholar]
  7. Embrapa. Business Guide for Artisanal Cheeses: Brazilian Artisanal Cheeses; Embrapa: Brasília, Brazil, 2021. [Google Scholar]
  8. Brazil. Ministry of Agriculture, Livestock and Food Supply. Technical Regulation on Hygienic-Sanitary Conditions and Good Manufacturing Practices for Food Processing/Manufacturing Establishments (Ordinance No. 368 of 4 September 1997). Off. Gaz. Union 1997, 1, 1. [Google Scholar]
  9. Brazil. Ministry of Agriculture, Livestock and Supply. Normative Instruction No. 53 of 16 October 2018. Technical Regulation of Identity and Quality of Milk Powder. Off. Gaz. Union 2018, 1, 11–12. [Google Scholar]
  10. Codex Alimentarius Comission. General Standard for Milk and Milk Products, 2nd ed.; FAO: Rome, Italy; OMS: Rome, Italy, 2011; ISSN 0259-2916. [Google Scholar]
  11. Oliveira, M.C.P. Protective Effect of Lactobacillus plantarum (B7) and L. rhamnosus (D1) from Artisanal Minas Cheese Against Experimental Escherichia coli EHEC and EIEC Infection and the Development of Fermented Buffalo Milk. Ph.D. Thesis, Federal University of Minas Gerais, Belo Horizonte, Brazil, 2016. [Google Scholar]
  12. Senz, M.; Keil, C.; Schmacht, M.; Palinski, S.; Cämmerer, B.; Hageböck, M. Influence of Media Heat Sterilization Process on Growth Performance of Representative Strains of the Genus Lactobacillus. Fermentation 2019, 5, 20. [Google Scholar] [CrossRef]
  13. Mendes, D.P.G. Physicochemical, Microbiological Characteristics and Sensory Acceptance of Fermented Milks by Lactic Acid Bacteria Isolated from Coalho Cheese from Pernambuco. Master’s Thesis, Federal University of Minas Gerais, Belo Horizonte, Brazil, 2011. [Google Scholar]
  14. NBR 12894; Sensory Analysis of Foods and Beverages—Classification. ABNT—Brazilian Association of Technical Standards: Rio de Janeiro, Brazil, 1994.
  15. Brazil. Ministry of Agriculture, Livestock and Supply. In Official Methods for Analysis of Animal Origin Products; MAPA: Brasília, Brazil, 2022. [Google Scholar]
  16. Brazil. Ministry of Agriculture, Livestock and Supply. Secretariat of Agricultural Defense. Department of Inspection of Animal Origin Products. Normative Instruction No. 46: Technical Regulation of Identity and Quality of Fermented Milks. Off. Gaz. Union 2007, 1, 5. [Google Scholar]
  17. IDF—International Dairy Federation. Yogurt: Enumeration of Characteristic Microorganisms Count Technique at 37 °C. Bull. IDF 1988, 117, 1–4. [Google Scholar]
  18. IDF—International Dairy Federation. Milk and Milk Products: Enumeration of Coliforms—Colony Count Technique and Most Probable Number Technique at 30 °C. Bull. IDF 1985, 73A, 1–8. [Google Scholar]
  19. APHA. Enumeration of Characteristic Microorganisms of Molds and Yeasts, 5 ed.; American Public Health Association: Washington, DC, USA, 2015; Chapter 21. [Google Scholar]
  20. Seleme, M.V.C.; Reckziegel, C.G.; Veitt, M.T.; Signor, A.; Palácio, S.M.; Gonçalves, G.C.; Barbieri, J.C.Z. Sensory Analysis of Caffeinated and Decaffeinated Coffea arabica L. of Artisanal Production and a Commercial Brand: Comparative Analysis between Coffees. Rev. Univ. Val. Rio Verde 2022, 21, 1. [Google Scholar]
  21. Üçok, G.; Sert, D. Growth Kinetics and Biomass Characteristics of Lactobacillus plantarum L14 Isolated from Natural Sourdough: Effect of Fermentation Time on Dough Machinability. LWT Food Sci. Technol. 2020, 129, 109516. [Google Scholar] [CrossRef]
  22. Lopes, D.S.; Bastos, P.A.M.B.; Rebello, L.P.G. Advances in the Application of Probiotic Lactic Acid Bacteria in Dairy Fermented Products: A Review. Vértices 2023, 25, 1. [Google Scholar] [CrossRef]
  23. Valente, G.L.C. In Vitro and In Vivo Probiotic Potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1 against Listeria monocytogenes in Experimental Infection and Goat Fermented Milk Production. Master’s Thesis, Federal University of Minas Gerais, Belo Horizonte, Brazil, 2018. [Google Scholar]
  24. Lunardi, A.; Dantas Filho, J.V.; Ferreira, C.C.; Cavali, J.; Vais, J.O.; Dias, A.A.; Gasparotto, P.H.G. Non-Starter Lactic Acid Bacteria (NSLAB): A Challenge to the Cheese Industry. Braz. J. Dev. 2021, 7, 26383–26409. [Google Scholar] [CrossRef]
  25. Ali, A.H.; Abu-Jdayil, B.; Al Nabulsi, A.; Osaili, T.; Liu, S.-Q.; Kamal-Eldin, A.; Mutamed, A. Effect of Acid Whey Protein Concentrate on the Rheological Properties, Antioxidant Capacities, and Biological Activities of Bioaccessible Fractions in Fermented Camel Milk. J. Dairy Sci. 2023, 108, 1242–1260. [Google Scholar]
  26. Vieira, A.C.A.; Andrade, E.H.P.; Soares, C.F.; Acurcio, L.B.; Mello, T.C.S.; de Souza, M.R. Probiotic Potential of Lactic Acid Bacteria Isolated from Artisanal Minas Cheese. Rev. Inst. Laticínios Cândido Tostes 2022, 77, 32–42. [Google Scholar] [CrossRef]
  27. Freitas, L.S.; Costa, W.P.G.; Oliveira, D.D. Microbiological Analysis of Sweets Sold by Delivery during the COVID-19 Isolation Period. Rev. Saúde Multidiscip. 2023, 14, 33–37. [Google Scholar]
  28. Carneiro, C.D.S.; Cunha, F.L.; Carvalho, L.D.; Carrijo, K.D.F.; Borges, A.; Cortez, M.A.S. Fermented Milks: History, Composition, Physicochemical Characteristics, Processing Technology and Defects. PUBVET 2012, 6, 1424. [Google Scholar]
  29. Francisquini, J.A.; Martins, E.; Silva, P.H.F.; Schuck, P.; Perrone, Í.T.; Carvalho, A.F. Maillard Reaction: A Review. Rev. Inst. Laticínios Cândido Tostes 2017, 72, 48–57. [Google Scholar] [CrossRef]
  30. Silva, M.A.P.; Leão, K.M.; Santos, P.A. Technology of Fermented Dairy Manufacturing: Bibliographic Review. PUBVET 2010, 4, 814. [Google Scholar]
  31. Bezerril, F.F. Development of Goat Yogurt Added with Xique-Xique Jelly (Pilosocereus gounellei): Evaluation of Technological Characteristics and Functional Potential. Ph.D. Thesis, Federal University of Paraíba, João Pessoa, Brazil, 2021. [Google Scholar]
  32. Brazil. National Health Surveillance Agency (ANVISA). Technical Regulation on Microbiological Standards for Foods (RDC No. 12 of 2 January 2001). Off. Gaz. Union 2001, 1, 1. [Google Scholar]
  33. Pereira, M.T.; Santana, E.H.W.; Santos, J.S. Importance of Lactic Acid and Non-Starter Bacteria (NSLAB) in Dairy Derivatives Production Technology. Ensaios 2020, 24, 348–352. [Google Scholar] [CrossRef]
  34. Dan, T.; Chen, H.; Li, T.; Tian, J.; Ren, W.; Zhang, H.; Sun, T. Influence of Lactobacillus plantarum P-8 on Fermented Milk Flavor and Storage Stability. Front. Microbiol. 2019, 9, 3133. [Google Scholar] [CrossRef] [PubMed]
Fermentation 11 00560 g001
Figure 1. Counts of L. casei B5 and L. plantarum B7 (log CFU/mL ± standard deviation) in different treatments during the storage period. According to Chart 1, TRAT1B5: Treatment 1 with L. casei; TRAT2B5: Treatment 2 with L. casei; TRAT3B5: Treatment 3 with L. casei and TRAT1B7: Treatment 1 1 with L. plantarum; TRAT2B7: Treatment 2 with L. plantarum; TRAT3B7: Treatment 3 with L. plantarum.
Figure 1. Counts of L. casei B5 and L. plantarum B7 (log CFU/mL ± standard deviation) in different treatments during the storage period. According to Chart 1, TRAT1B5: Treatment 1 with L. casei; TRAT2B5: Treatment 2 with L. casei; TRAT3B5: Treatment 3 with L. casei and TRAT1B7: Treatment 1 1 with L. plantarum; TRAT2B7: Treatment 2 with L. plantarum; TRAT3B7: Treatment 3 with L. plantarum.
Fermentation 11 00560 g001
Fermentation 11 00560 ch001
Chart 1. Codes used for identifying treatments and corresponding bacterial strains.
Chart 1. Codes used for identifying treatments and corresponding bacterial strains.
Fermentation 11 00560 ch001
Table 1. Means of pH values and standard deviation for fermented milks using L. casei B5 and L. plantarum B7 during the storage period under refrigeration.
Table 1. Means of pH values and standard deviation for fermented milks using L. casei B5 and L. plantarum B7 during the storage period under refrigeration.
TRAT1B5TRAT2B5TRAT3B5TRAT1B7TRAT2B7TRAT3B7
DAY 14.15 ± 0.01 A,a3.95 ± 0.02 A,c3.95 ± 0.01 A,c3.76 ± 0.02 A,d4.02 ± 0.02 A,b3.98 ± 0.00 A,bc
DAY 154.02 ± 0.01 B,a3.87 ± 0.01 B,c3.93 ± 0.01 A,b3.76± 0.01 A,d3.88 ± 0.01 B,c3.92 ± 0.01 B,b
DAY 303.84 ± 0.01 C,a3.65 ± 0.00 C,d3.82 ± 0.00 B,a3.57 ± 0.01 B,e3.71 ± 0.01 C,c3.79 ± 0.00 D,b
DAY 453.57 ± 0.02 D,de3.64 ± 0.06 C,c3.82 ± 0.02 B,a3.55 ± 0.03 B,e3.70 ± 0.03 C,bc3.83 ± 0.03 C,a
Legend: Uppercase letters (A, B, C, D) indicate statistically significant differences (p < 0.05) between treatments. Lowercase letters (a, b, c, d, e) indicate statistically significant differences (p < 0.05) between storage days.
Table 2. Means of titratable acidity values expressed as g of lactic acid/100 g and their standard deviations for fermented milks using L. casei B5 and L. plantarum B7 each treatment during the storage period under refrigeration.
Table 2. Means of titratable acidity values expressed as g of lactic acid/100 g and their standard deviations for fermented milks using L. casei B5 and L. plantarum B7 each treatment during the storage period under refrigeration.
TRAT1B5TRAT2B5TRAT3B5TRAT1B7TRAT2B7TRAT3B7
DAY 11.17 ± 0.01 AB,e1.52 ± 0.02 C,b1.27 ± 0.00 C,d1.46 ± 0.02 A,c1.60 ± 0.02 A,a1.19 ± 0.02 A,e
DAY 151.33 ± 0.17 A,abc1.63 ± 0.15 BC,a1.45 ± 0.29 ABC,ab1.32 ± 0.08 B,abc1.52 ± 0.04 B,ab1.07 ± 0.04 C,b
DAY 301.21 ± 0.03 AB,e1.81 ± 0.02 AB,a1.73 ± 0.02 A,b1.34 ± 0.00 B,d1.66 ± 0.03 A,c1.09 ± 0.01 B,f
DAY 451.14 ± 0.00 B,e1.78 ± 0.07 B,a1.69 ± 0.02 AB,b1.35 ± 0.01 B,d1.64 ± 0.03 A,c1.11 ± 0.02 B,e
Legend: Uppercase letters (A, B and C)indicate statistically significant differences (p < 0.05) between treatments. Lowercase letters (a, b, c, d, e, f) indicate statistically significant differences (p < 0.05) between storage days.
Table 3. Means counts of L. casei B5 and L. plantarum B7 (log CFU/mL), their standard deviations in fermented milks during the storage period.
Table 3. Means counts of L. casei B5 and L. plantarum B7 (log CFU/mL), their standard deviations in fermented milks during the storage period.
TRAT1B5TRAT2B5TRAT3B5TRAT1B7TRAT2B7TRAT3B7
DAY 18.35 ± 0.70 A,a8.49 ± 0.12 A,a8.39 ± 0.28 A,a8.29 ± 0.04 A,a7.78 ±0.01 A,a8.22 ± 0.19 A,a
DAY 159.27 ± 0.05 A,a7.63 ± 0.28 B,c8.18 ± 0.08 B,b8.15 ± 0.02 B,b8.43 ± 0.01 B,b8.40 ±0.05 A,b
DAY 308.26 ±0.03 A,ab7.59 ± 0.19 B,c8.47 ± 0.14 A,a8.46 ± 0.01 C,a10.47 ± 0.01 C,a8.12 ± 0.0 A,b
DAY 458.32 ±0.35 A,a8.14 ± 0.14 AB,c8.46 ± 0.01 A,b8.43 ± 0.21 C,b8.45 ± 0.02 B,b8.47 ± 0.0 A,b
Legend: Uppercase letters (A, B, and C) indicate statistically significant differences (p < 0.05) between treatments. Lowercase letters (a, b, and c) indicate statistically significant differences (p < 0.05) between storage days.
Table 4. Results of the preference ranking test for different fermented milk treatments, evaluated by the Friedman Test.
Table 4. Results of the preference ranking test for different fermented milk treatments, evaluated by the Friedman Test.
Preference Ranking
TRAT3B7TRAT2B7TRAT2B5TRAT3B5TRAT1B7TRAT1B5
TRAT3B7397283256245222181
TRAT2B7-114141152175216
TRAT2B5--273861102
TRAT3B5---113475
TRAT1B7----2364
TRAT1B5-----41
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Queiroz, C.S.d.; Rekowsky, B.S.d.S.; Salgado, M.J.G.; Lopes, M.F.B.d.H.; Souza, M.R.d.; Costa, M.P.d.; Silva, J.G.d. Development of Fermented Milks with Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 Isolated from Minas Artisanal Cheese. Fermentation 2025, 11, 560. https://doi.org/10.3390/fermentation11100560

AMA Style

Queiroz CSd, Rekowsky BSdS, Salgado MJG, Lopes MFBdH, Souza MRd, Costa MPd, Silva JGd. Development of Fermented Milks with Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 Isolated from Minas Artisanal Cheese. Fermentation. 2025; 11(10):560. https://doi.org/10.3390/fermentation11100560

Chicago/Turabian Style

Queiroz, Camila Selles de, Bruna Samara dos Santos Rekowsky, Madian Johel Galo Salgado, Maria Fernanda Barreto da Hora Lopes, Marcelo Resende de Souza, Marion Pereira da Costa, and José Givanildo da Silva. 2025. "Development of Fermented Milks with Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 Isolated from Minas Artisanal Cheese" Fermentation 11, no. 10: 560. https://doi.org/10.3390/fermentation11100560

APA Style

Queiroz, C. S. d., Rekowsky, B. S. d. S., Salgado, M. J. G., Lopes, M. F. B. d. H., Souza, M. R. d., Costa, M. P. d., & Silva, J. G. d. (2025). Development of Fermented Milks with Lacticaseibacillus casei B5 and Lactiplantibacillus plantarum B7 Isolated from Minas Artisanal Cheese. Fermentation, 11(10), 560. https://doi.org/10.3390/fermentation11100560

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