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Article

Impacts of Sonication on Fermentation Process and Physicochemical, Microbiological and Sensorial Characteristics of Fermented Black Carrot Juice

by
Muhammet Ercan
1,
Mehmet Akbulut
2,*,
Hacer Çoklar
2 and
Talha Demirci
2
1
Department of Food Engineering, Graduate School of Natural and Applied Sciences, Bingol University, Bingol 12000, Türkiye
2
Department of Food Engineering, Agriculture Faculty, Selcuk University, Konya 42130, Türkiye
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(8), 475; https://doi.org/10.3390/fermentation11080475
Submission received: 24 July 2025 / Revised: 11 August 2025 / Accepted: 13 August 2025 / Published: 19 August 2025
(This article belongs to the Special Issue 10th Anniversary of Fermentation: Feature Papers in the "Fermentation for Food and Beverages" Section)
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Abstract

In recent years, ultrasound has been integrated into fermentation technology due to its activating effect on microorganisms, and the possible effects of ultrasound-assisted fermentation on the fermentation process, yield and quality of the final product have also attracted attention. This study aimed to reveal the effects of sonication applied before the fermentation on the fermentation process and the quality of fermented black carrot juice. The samples were sonicated at a frequency of 35 kHz and an amplitude of 60% for 0, 5, 15 or 30 min before the fermentation. During the fermentation, the pH, acidity, organic acid profile, ethanol and soluble solid content (SSC), color, turbidity, total lactic acid bacteria (LAB), total mesophilic aerobic bacteria (TMAB) and yeast counts were determined. The amount of SSC in the samples increased at the beginning of fermentation as the sonication time increased. Lactic, acetic and propionic acids were detected in the samples. The amount of lactic acid in all the samples treated with ultrasound was higher than in the control sample and the amounts of acetic acid, propionic acid and ethanol were lower. Ultrasound application caused an increase in the TMAB and yeast counts. A five-minute ultrasound application caused a decrease in the number of LAB, while 15- and 30-min applications caused an increase. Thirty minutes of ultrasound treatment resulted in the reddest fermented black carrot juices with the highest level of color saturation. The most appreciated sample in terms of taste, aroma and general acceptability was the sample subjected to a five-minute ultrasound application. As a result, ultrasound application before fermentation positively supports different quality parameters of fermented black carrot juice and the use of sonication in production can be recommended.

1. Introduction

Ultrasound is defined as sound waves with a frequency above the hearing limit of the human ear (20 kHz and above). It is also referred to as high-frequency pressure waves. Ultrasound within the 20 kHz−1 MHz range is characterized as low-frequency or high-power sound waves. Ultrasonic waves with a frequency greater than 1 MHz are classified as high-frequency sound waves and are frequently referred to as diagnostic sound waves [1]. Ultrasound technology finds application in a variety of fields such as enzyme inhibition, microorganism inactivation, foaming, foam breaking, improving filtration efficiency, crystallization of fats and sugars, CO2 degassing during fermentation, maturation and oxidation of alcoholic beverages, meat aging, humidification, cutting, waste treatment, precipitation of airborne dust, blending, freezing, melting, surface decontamination, emulsification and extraction of bioactive components [2,3,4,5,6,7,8,9,10,11,12]. In addition to microbial inactivation, ultrasound can also stimulate cell growth [13]. The use of appropriate ultrasound conditions favorably enhances the growth of microbial cells. Low-intensity sound waves create stable cavitations in liquid media and cause reversible damage to microbial cells. In this way, it accelerates the growth of microbial cells, changes their life phases and enables them to produce more metabolic products.
In previous studies, it has been reported that ultrasound application increases fibrinolytic enzyme production from Bacillus sphaericus MTCC 3672 by 1.48 times [14], accelerates the proliferation mechanism of Saccharomyces cerevisiae cells [15], shortens the fermentation time by up to 24 h and increases the amount of ethanol by 3.5 times [16,17], shortens fermentation time up to 36 h, increases the amount of riboflavin by 5 times in the production of riboflavin from Ecemothecium ashbyii [18], reduces the fermentation time of beverages such as wine, beer and sake produced with S. cerevisia by 50-64% [19] and reduces fermentation time in yogurt production by approximately 40% [20,21].
Although the mechanism by which ultrasound promotes the growth of microorganisms is not fully understood, it has been suggested that ultrasound provides faster passage of components, such as sugars, from plant tissue to the fermentation medium, increases cell biomass by improving the nutrient utilization of bacterial cells, accelerates intracellular and extracellular substance transfer due to increased cell membrane permeability, removes toxic compounds from the cell, and accelerates the entry of nutrients into the cell.
Fermented black carrot juice is a red-colored, cloudy and sour-tasting fermented lactic beverage. Its production involves the use of bulgur flour, turnip, water, sourdough, black carrot and salt as raw materials [22,23]. The beverage has its origins in the southeastern Anatolian provinces of Türkiye, particularly Adana, Mersin, Hatay and Kahramanmaras, and has become a well-known and popular beverage throughout Turkey [24]. The beverage has been reported to exhibit antiproliferative activity and probiotic properties thanks to its phenolic compounds and microbial flora and has been reported to have positive effects on health [25,26]. Fermented black carrot juice contains S. cerevisiae, non-Saccharomyces yeasts, and both homo and heterofermentative lactic acid bacteria. The predominant genus among the lactic acid bacteria is Lactobacillus, comprising 89.63%, followed by Leuconostoc (9.63%) and Pediococcus (0.74%) [22]. The sugar necessary for the development of lactic acid bacteria is found in raw materials.
The fermentation process of black carrot juice typically requires 20 to 45 days, with the duration contingent on factors such as the chosen fermentation method and the ambient temperature. Although it is thought that fermentation is completed when acidity development stops in lactic acid fermentation, it is known that fresh fermented beverages are sensory-weak and need post-fermentation maturation for taste, aroma, color and texture development [27,28]. During maturation, a variety of biochemical reactions take place, leading to the formation of compounds that contribute to the development of color and aroma. Ultrasound is of great interest in fermentation technology as it promotes the growth of microorganisms, increases enzyme activity and metabolic performance, rapidly supplies the microorganism with the components it needs through extraction and enables post-fermentation maturation by promoting reactions such as oxidation, esterification, proteolysis and the Maillard reaction during maturation [27,28,29,30].
To the best of our knowledge, no study has been conducted on the effect of ultrasound applied to fermented black carrot juice before fermentation. This research was designed to provide a new source for literature and to provide data for industrial production. The aim of this study was to determine the effects of ultrasound applications at different times before the fermentation on the fermentation process of fermented black carrot juice, as well as on the beverage’s color, acidity, ethyl alcohol content, turbidity, total bacteria count, lactic acid bacteria count, yeast count and sensory properties.

2. Materials and Methods

2.1. Material

Black carrots, bulgur flour, baker’s yeast (Saccharomyces cerevisiae), salt and drinking water were used in the production of fermented black carrot juice. Black carrots and bulgur flour were obtained from Gunseven Inc., Ereğli, Türkiye. Baker’s yeast (Pak Gida, Düzce, Türkiye), salt (Billur Salt Industry Co., İzmir, Türkiye) and water (Buzdağı, Seli Inc., Sakarya, Türkiye) were purchased from the local market.

2.2. Methods

2.2.1. Fermentation and Sonication Methods

Bulgur (0.91%) and yeast (0.2%) were subjected to fermentation in a 1 L glass jar at ambient temperature for a 24-h period. Subsequently, black carrots (16.6%), which had been sliced into 1 cm thick and 10 cm long pieces after the removal of their heads and tails, salt (1.16%), and water were added. The jars were tightly closed. In this way, 24 samples with the same formulation were prepared and divided into 4 groups with 6 samples in each group. The initial group of six samples was not exposed to ultrasound, and these samples were considered the control group. The second group subjected to the ultrasound procedure was exposed for a duration of five minutes, while the third group was exposed for 15 min and the fourth group was exposed for 30 min. On the first day, a single jar from each group was selected for analysis. The remaining jars were subjected to a fermentation process at ambient temperature (25 °C). The fermentation process was periodically evaluated by analyzing samples collected from each group on days 4, 8, 12, 20 and 30. The ultrasound application was performed in an ultrasonic-assisted water bath (Elma, Transsonic TI-H-10, Singen, Germany). The jars, tightly sealed, were placed in the device and exposed to sonication at a frequency of 35 kHz and an amplitude of 60%. During the sonication process, the temperature was controlled and maintained at 25 °C by means of water circulation. The experiment was carried out in 3 replicates.

2.2.2. Analysis Methods

pH, Soluble Solid Content, Total Titratable Acidity, Color and Turbidity Analyses
The pH of the fermented carrot juice samples was measured using a glass electrode pH meter (Inolab 720, WTW, Weilheim, Germany), while the soluble solid content values were determined with an Atago refractometer (HSR-500, Tokyo, Japan). The turbidity values were measured with a WTW turbidimeter (TURB 430 T, Weilheim, Germany), and the reflectance color values (L*, a*, b*, C*, and h) of the samples were measured with a spectrophotometer (CM-5, Konica Minolta, Osaka, Japan). All measurements were carried out at 20 °C. For the total titratable acidity analysis, 10 mL of the sample was taken, 20 mL of the pure water was added, and the pH was determined by titrating with 0.1 N NaOH until the pH was 8.2. The results are expressed as g lactic acid equivalent/100 mL [31].
Organic Acid Profile and Ethyl Alcohol Analyses
Organic acid profile and ethyl alcohol content of fermented black carrot juices were determined by HPLC (Agilent 1260, Agilent Technologies, Inc., Santa Clara, CA, USA). Prior to injection, the samples were diluted 1:2 (v/v) with ultrapure water and passed through a 0.45 μm pore diameter PVDF (polyvinylidene fluoride) filter. Separation was carried out on an Aminex HPX-87H column. Sulfuric acid (0.005 N) was used as mobile phase, and the flow rate was set to 0.6 mL/min. Ethanol was detected with a refractive index detector and organic acids were detected with a DAD detector set to 210 nm wavelength [32].
Microbiological Analyses
Plate Count Agar was used for total mesophilic aerobic bacteria count, Potato Dextrose Agar for yeast count and De Man, Rogosa and Sharpe agar for lactic acid bacteria (LAB) count. Dilutions up to 10−10 were prepared from fermented black carrot juices and 1 mL was taken to the media and inoculated according to the pour plating method. The inoculated petri dishes were incubated at 37 °C for 48 h for total mesophilic bacteria count, at 28 °C for 72 h for yeast count and at 37 °C for 48 h for lactic acid bacteria count. Microbiological counting results of fermented black carrot juices were given as log cfu/mL [23].
Sensory Evaluation
Sensory analysis of fermented black carrot juice was carried out using a 0- to 9-point scale indicating color, aroma, taste, appearance and overall acceptability. The sensory analysis was conducted in accordance with ISO standard guidelines with a group of 15 volunteer panelists consisting of male and female non-smokers, aged between 25 and 50 years, from the Faculty of Agriculture, Selcuk University. The panelists conducted sensory evaluations of the samples taken on the 12th, 20th and 30th day of the fermentation period. The samples were presented to the panelists in 50 mL transparent glasses at 4 °C temperature [23].
Statistical Analysis
The results were given as mean ± standard deviation. To determine the effect of both ultrasound application and fermentation process, the data obtained were subjected to Analysis of Variance using MINITAB release 16.0 program. The Tukey Multiple Comparison Test was used to check whether the differences between group means were significant.

3. Results

Table 1 shows the pH, total titratable acidity, turbidity and soluble solid content values in the fermentation process of fermented black carrot juice samples. At the beginning of fermentation, the pH of the samples ranged from 5.85 to 6.17 and it was seen that the pH increased after 5 and 15 min of ultrasound application and the pH decreased with a 30 min ultrasound application. However, on the 30th day of fermentation, the lowest pH value was determined in the samples with ultrasound applied for 15 min.
The soluble solid content of the fermented black carrot juices ranged from 1.40 to 1.75 at the beginning of fermentation and from 1.75 to 2.25 at the end of fermentation. The soluble solid content increased with 15- and 30-min ultrasound applications before fermentation, and the soluble solid content of these samples was lower during the fermentation.
During the fermentation, the pH decreased, and the acidity and soluble solid content increased until the 30th day, although it showed fluctuations. Although the fermentation process had a statistically significant effect on the pH, acidity, turbidity, and soluble solid content, the effect of sonication on these parameters was statistically insignificant.
The organic acid content of fermented black carrot juice samples is illustrated in Figure 1, while the ethanol content is demonstrated in Figure 2. Lactic, acetic and propionic acids were detected in the samples. At the beginning of fermentation, the control sample had the lowest lactic acid content (0.110 mg/100 mL), while the sample treated with ultrasound for 15 min had the highest content (0.184 mg/100 mL). At the beginning of fermentation, the lowest lactic acid content was observed in the control sample (0.110 mg/100 mL), while the highest content was recorded in the sample exposed to 15 min of ultrasound (0.184 mg/100 mL). The highest amount of lactic acid was determined on the 20th day of fermentation in the control sample to be in the 5-min ultrasound-treated samples, and on the 30th day of fermentation in the 15-min ultrasound-treated sample. After 30 days of fermentation, it was found that the samples treated with ultrasound had a higher level of lactic acid compared to the control sample.
As in the case of lactic acid, it was determined that the amounts of acetic acid, propionic acid and ethanol in the samples treated with ultrasound at the beginning of fermentation were lower. As the fermentation process continued, a decrease in the levels of acetic and propionic acids were observed in the samples treated ultrasound. After 30 days of the fermentation, the lowest propionic acid was found in the samples that had been treated with 30 min of ultrasound, and the lowest acetic acid was found in the samples that had been exposed to 5 and 15 min of ultrasound. As the fermentation process approached its 30th day, a higher concentration of ethanol was observed in the sample subjected to 5 min of ultrasound treatment than that of the control sample. Conversely, a lower concentration of ethanol was detected in the samples that were exposed to 15 and 30 min of ultrasound treatment, in contrast to the control sample.
Table 2 shows the total mesophilic aerobic bacteria, lactic acid bacteria and yeast count of fermented black carrot juices. The ultrasound treatment exhibited a significant impact on the microbial load and population within the fermentation medium. At the beginning of fermentation, an increase in the total number of bacteria and yeast was observed with ultrasound application, while ultrasound application caused a decrease in the number of lactic acid bacteria, except for 30 min of ultrasound application. As the fermentation progressed, the total number of bacteria decreased. This decrease was the lowest in the control sample and the highest in the sample treated with ultrasound for 5 min. During the fermentation, there was an increase on the 8th day and a decrease on the 12th day.
The lactic acid bacteria count was higher in the control sample than in the sample treated with ultrasound for 5 min, and lower than in the samples treated with ultrasound for 15 or 30 min. On the 12th day of fermentation, the highest lactic acid bacteria count was detected in the sample treated with ultrasound for 30 min. The yeast count decreased as the fermentation progressed. The lowest value was found in the control sample, while the highest value was found in the sample that was treated with ultrasound for 30 min. The lactic acid bacteria count of the control sample was higher than that of the sample treated with ultrasound for 5 min, and lower than the samples treated with ultrasound for 15 or 30 min. On the 12th day of fermentation, the highest lactic acid bacteria count was detected in the sample treated with ultrasound for 30 min. The yeast count decreased as the fermentation progressed. The lowest value was found in the control sample, while the highest value was found in the sample that was treated with ultrasound for 30 min.
Table 3 presents the color values of fermented black carrot juice samples. The L*, a*, b*, C* and h represent brightness, redness, yellowness, color saturation and angle value, respectively, on the Hunter scale.
At the initiation of fermentation, the L*, a*, b*, C* and h values of the control sample were 70.8, 0.78, 5.02, 5.08 and 81.16, respectively. Throughout the fermentation, the brightness (L*) and h values of the fermented black carrot juices decreased, while the redness (a*), yellowness (b*) and color saturation (C*) increased. At the beginning of fermentation, the L* and h values of the samples decreased with ultrasound application, the redness values of the samples with 15- and 30-min ultrasound application increased and the yellowness and color saturation values of the samples with 30 min of ultrasound application were higher than the control sample. It was found that the reddest and most saturated color was obtained on the 30th day of fermentation of the sample treated with ultrasound for 30 min.
Figure 3 shows the radar plot of the sensory evaluation of the fermented black carrot juice samples on days 12, 20 and 30 of fermentation. On the 12th day of the fermentation, the sample that received a 30-min ultrasound treatment had the lowest sensory scores in terms of taste, appearance and overall acceptability. In terms of color, aroma and overall acceptability, the samples with 5 and 15 min of ultrasound application scored higher than the control sample. On the 20th day of fermentation, the ultrasound-applied samples were more appreciated in terms of all sensory parameters except color. The panelists rated the fermented black carrot juice produced with a 5-min ultrasound application as the most acceptable in terms of taste, aroma and overall acceptability. In terms of appearance, the sample treated with 30 min of ultrasound was the most highly appreciated. In terms of color, the sample without ultrasound treatment was most highly rated. Ultrasound treatment had an overall positive effect on the sensory characteristics of the fermented black carrot juices.

4. Discussion

Fermented black carrot juice is a red-colored, cloudy beverage that is produced through the fermentation of black carrot, bulgur flour and baker’s yeast in a brine solution at room temperature for a period of 2–4 weeks. The total titratable acidity ranges from 0.11% to 0.91%, and the pH ranges from 3.15 to 4.25 [24,33]. In the fermentation of fermented black carrot juice, S. cerevisiae, added to the medium, and natural microbiota, predominantly lactic acid bacteria originating from the raw materials utilized, play a crucial role. The amount of lactic acid bacteria in the fermented black carrot juice was found to range between 4.08 and 7.53 log cfu/mL, the total number of bacteria between 4.20 and 9.06 log cfu/mL, and the number of yeasts between 2.1 and 6.70 log cfu/mL [23,31,32,34]. The physicochemical, sensory and microbiological properties of the fermented black carrot exhibit variability depending on the fermentation conditions and the composition and properties of the raw materials utilized. In this study, the properties of the product obtained by sonication application vary among themselves but are within the range of values specified in the literature.
Sonication is an effective method of extracting bioactive compounds, particularly water-soluble components, from plant tissues [35,36]. During the fermentation of plant products, compounds within the tissue are transferred out of the cell by diffusion. The increase in the soluble solid content at the beginning of fermentation with sonication application may be attributed to the fact that the substance transfer, which should be by diffusion, is accelerated by sonication-assisted extraction. The fluctuation in the soluble solid content during the fermentation may be attributed to the transfer of water-soluble components from the raw materials to the medium, the breakdown of polymeric components into smaller building blocks due to the enzymatic activity, the transfer of components such as salt in the fermentation medium to the raw materials, the utilization of components in the medium by microorganisms and/or the production of new components [37]. Although statistically insignificant, the increase in pH at low sonication application times and the decrease at long application times may be attributed to the migration of components such as salt into solid materials, especially carrots, and the free amino acids, pectic compounds and organic acids found in the structure of black carrots into the medium.
During the fermentation process of black carrot juice, a decrease in the pH and an increase in the acidity were observed. The decrease in the pH and the increase in the acidity can be attributed to the presence of organic acids, which are formed because of sugars being utilized within the environment due to microbial activity. The sugar required for the growth of lactic acid bacteria in fermented black carrot juice is found in the raw materials, especially in carrots and bulgur flour. The easier access of microorganisms to the components passing from tissues and cells to the fermentation liquid by sonication may have supported the growth of microorganisms.
S. cerevisiae and non-Saccharomyces yeasts (P. kudriavzevii, P. fermentans, Candida oleophila, Kazachstania bulderi and Geotrichum candidum) [34], as well as homofermentative and heterofermentative lactic acid bacteria (Lactobacillus plantarum, L. paracasei subsp. paracasei, L. brevis, L. fermentum, Leu.mesenteroides subsp. mesenteroid, P. pentosaceaceus and L. delbrueckii subsp. delbrueckii) [38] are involved in the fermentation process of fermented black carrot juice.
Lactic acid bacteria are classified as either homolactic or heterolactic. Homofermentative bacteria produce lactic acid from fermentable sugars, while heterofermentative bacteria produce secondary products, such as ethanol, acetic acid, diacetyl and carbon dioxide, in addition to lactic acid. Yeasts are capable of metabolizing fermentable sugars to alcohol under anaerobic conditions.
The sonication decreased the amount of lactic acid bacteria. However, the 30-min application did not differ statistically from the control sample. The sample sonicated for 5 min had a lower lactic acid bacteria count than the control sample both at the beginning of the fermentation and throughout fermentation, while the lactic acid bacteria count of the other ultrasound-applied samples was higher than the control sample. The 5-min sonication treatment of the samples may have exhibited an inhibitory effect on lactic acid bacteria, while the applications for 15 and 30 min may have had an activating effect. The higher levels of acetic and propionic acids in the sonicated samples could be due to the activation of heterofermentative lactic acid bacteria. The higher amount of ethyl alcohol in the initial stages of fermentation compared to the control sample may be attributed to the fact that yeast and heterofermentative bacteria are more active in the fermentation environment, as supported by the yeast and lactic acid bacteria numbers. The total bacterial count increased after the ultrasound application. However, throughout the fermentation, the count was lower in the sonicated samples than in the control sample.
The increase in fermentation efficiency is attributed to the stimulation of microorganisms with ultrasound and the consequent improvement of their metabolism [39]. In previous studies, it has been reported that the application of ultrasound shortened fermentation time, increased yield and accelerated cell proliferation in both alcohol fermentation with S. cerevisiae [15,19] and lactic acid fermentation with lactic acid bacteria [21,40].
The sensitivity of microorganisms to ultrasound varies according to their structure. It is noted that Gram-negative bacteria are more sensitive to ultrasonic cavitation than Gram-positives due to the weak structure of their cell membranes, and bacilli are more sensitive to ultrasonic cavitation than cocci due to the more superficial cell walls [13]. The acidity of the environment, the formation of ethyl alcohol, the diffusion of antioxidant components from tissues into the environment and the resistance of microorganisms to these components affect yeast and bacteria. The application of sonication may have affected the resistance of yeast and lactic acid bacteria. The presence of lower levels of propionic acid and acetic acid and higher levels of lactic acid in the sonicated samples compared to the control sample suggests that sonication may have activated homofermentative lactic acid bacteria while inhibiting heterofermentative bacteria.
There are antagonistic and synergistic effects among S. cerevisiae, non-Saccharomyces yeast and homo and heterofermentative lactic acid bacteria. These interactions are basically attributed to the competition that occurs due to the consumption of the same nutrients by the microorganisms and the metabolites they produce supporting or inhibiting the growth of each other. In low-acid fermented foods, heterofermentative lactic acid bacteria increase the growth of S. cerevisiae, while S. cerevisiae generally inhibits the growth of heterofermentative lactic acid bacteria in higher alcohol environments. It has been shown that Pichia enhances heterofermentative lactic acid fermentation in a low-ethanol environment and increases the production of organic acids such as propionic acid and promotes lactic acid bacteria growth, while in a high-ethanol environment, it inhibits lactic acid bacteria growth and metabolism [41]. It is known that metabolites with antibacterial activity such as organic acids produced by heterofermentative lactic acid bacteria inhibit the growth and reproduction of non-Saccharomyces yeasts by secreting them. Since the sonication treatment showed activating or inactivating effects on S. cerevisiae, non-Saccharomyces, and homofermentative and heterofermentative lactic acid bacteria, it may have changed the population of these microorganisms and eliminated their antagonistic and synergistic interactions [41]. This approach may provide a possible explanation for the observed increase in lactic acid bacteria and yeast populations. Sonication affects enzyme activity and has been reported to increase the activity of certain enzymes. Under the right conditions, ultrasound can alter the structure of enzymes, speed up their contact with substrates, and enhance enzyme–substrate interactions, resulting in higher enzymatic activity [13].
In a previous study, 60 aroma compounds including 20 terpenes, 9 esters, 9 alcohols, 5 volatile acids, 6 volatile phenols, 5 lactones, 3 naphthalene, 2 carbonyl compounds and 1 C13-norisoprenoid have been detected in fermented black carrot juice [42]. The taste and aroma of the fermented black carrot juice originate from the raw materials used and fermentation and are additionally influenced by processing and storage conditions [42,43]. It has been reported that total aroma compounds decrease with pasteurization and storage [44]. The type of lactic acid bacteria in black carrot juice fermentation also causes changes in the flavor profile [42]. Fresh fermented foods that have just completed fermentation are sensorially weak, so they usually require a long maturation in terms of taste, aroma and color development. During the maturation of fermented foods, fermentation compounds that improve the sensory properties of fermented foods are formed through various biochemical reactions, primarily the Maillard reaction, esterification and proteolysis. It has been reported that acoustic cavitation in liquids with ultrasound application initiates these reactions and increases the reaction rates [28,39]. The acceleration of these reactions, as well as the differences in microbial diversity caused by sonication, may have affected the sensory properties of sonicated fermented black carrot juices. The a* value, which is indicative of redness, is derived from the black carrots present in the formulation. The observation that the a* value was lower in the samples sonicated at the beginning of fermentation than in the control sample can be attributed to the rapid transfer of anthocyanins from the carrots to the fermentation medium.
In general, it was observed that the sonicated samples were more turbid than the control sample at the beginning of fermentation and at the end of 30 days of fermentation. In a study, it was reported that ultrasound applied after fermentation increased the turbidity of fermented black carrot juice [32].
The components in a suspension, such as cellulose, protein, lipid, pectin and hemicellulose, cause turbidity. The increase in turbidity due to ultrasound application may be due to microbial enzyme activity, extraction of turbidity-causing components from the tissue, colloidal disintegration and fragmentation of macromolecules into small pieces. The fluctuations that occur during fermentation can be attributed to the precipitation of the components causing turbidity by forming complexes and the simultaneous occurrence of the above-mentioned colloidal disintegration and release from the tissue [31].

5. Conclusions

In conclusion, applying ultrasound before the fermentation significantly affected the microbiological and sensory properties of the fermented black carrot juice. The ultrasound treatment increased the number of TMAB and yeast. The number of lactic acid bacteria differed according to the application time. A five-minute ultrasound application caused a decrease in the number of LAB, while 15 and 30 min applications caused an increase. The sensory properties of the beverage were found to develop faster with the ultrasound treatment. Taste, aroma and color development were observed to be significantly complete in the ultrasound-treated samples on the 20th day of fermentation. The ultrasound application before the fermentation was found to positively support the sensory, color and microbiological properties of the fermented black carrot juice, as well as accelerate the development of sensory properties. Therefore, sonication is recommended for producing the fermented black carrot juice, particularly to improve sensory characteristics and to accelerate maturation.

Author Contributions

Conceptualization, M.A. and H.Ç.; methodology, M.A. and H.Ç.; software, M.E.; validation, M.A., H.Ç. and M.E.; formal analysis, M.E. and T.D.; investigation, M.E. and H.Ç.; resources, M.A.; data curation, M.E.; writing—original draft preparation, H.Ç.; writing—review and editing, M.A.; visualization, M.E. and H.Ç.; supervision, M.A.; project administration, M.A.; funding acquisition, M.A. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Teaching Staff Training Program, ÖYP-2015-083.

Institutional Review Board Statement

The national laws do not require ethical approval for sensory evaluation. There are no human ethics committees formal documentation procedures available for sensory evaluation. The experimental protocol involving sensory evaluation was in accordance with the relevant operation specifications in Turkey.

Informed Consent Statement

Written informed consent for participation was obtained from all subjects involved in the study, in accordance with the General Data Protection Regulation (GDPR) 2016/679. The study was conducted following the principles of the Declaration of Helsinki.

Data Availability Statement

The original contributions presented in the study are included in the article, and further inquiries can be directed to the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest.

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Fermentation 11 00475 g001aFermentation 11 00475 g001b
Figure 1. Lactic, acetic and propionic acid contents (g/100 mL) of fermented black carrot juice samples.
Figure 1. Lactic, acetic and propionic acid contents (g/100 mL) of fermented black carrot juice samples.
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Fermentation 11 00475 g002
Figure 2. Ethanol contents of fermented black carrot juice samples.
Figure 2. Ethanol contents of fermented black carrot juice samples.
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Fermentation 11 00475 g003aFermentation 11 00475 g003b
Figure 3. Radar plot of sensory evaluation of fermented black carrot juice samples.
Figure 3. Radar plot of sensory evaluation of fermented black carrot juice samples.
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Table 1. pH, total titratable acidity, soluble solid content and turbidity values of fermented black carrot juice samples.
Table 1. pH, total titratable acidity, soluble solid content and turbidity values of fermented black carrot juice samples.
Sonication Time (min)Fermentation Period (Day)pHTotal Titratable Acidity (g/L)Soluble Solid Content (%)Turbidity
(NTU)
005.85 ± 0.090.011 ± 0.0001.40 ± 0.28163.00 ± 12.73
43.72 ± 0.050.207 ± 0.0032.00 ± 0.00255.00 ± 0.00
83.44 ± 0.020.377 ± 0.0182.40 ± 0.28208.50 ± 31.80
123.52 ± 0.040.301 ± 0.0181.80 ± 0.00121.30 ± 74.60
203.36 ± 0.090.378 ± 0.09122.15 ± 0.21223.50 ± 77.10
303.53 ± 0.000.359 ± 0.0182.25 ± 0.21162.50 ± 0.71
506.17 ± 0.450.008 ± 0.0051.40 ± 0.00181.00 ± 17.00
43.61 ± 0.100.205 ± 0.0872.00 ± 0.00271.00 ± 45.30
83.49 ± 0.020.268 ± 0.0541.80 ± 0.28183.50 ± 2.12
123.48 ± 0.010.292 ± 0.0661.70 ± 0.42167.50 ± 37.50
203.38 ± 0.010.407 ± 0.0262.20 ± 0.00217.00 ± 7.07
303.50 ± 0.050.353 ± 0.0612.00 ± 0.57190.00 ± 80.60
1506.05 ± 0.200.006 ± 0.0001.70 ± 0.14221.50 ± 7.78
43.62 ± 0.020.211 ± 0.0482.05 ± 0.07271.00 ± 58.00
83.44 ± 0.020.364 ± 0.0262.40 ± 0.00157.50 ± 40.30
123.45 ± 0.000.288 ± 0.0361.65 ± 0.21208.50 ± 26.20
203.49 ± 0.060.299 ± 0.0511.85 ± 0.50219.00 ± 65.10
303.43 ± 0.010.420 ± 0.0591.75 ± 0.35148.00 ± 35.40
3005.83 ± 0.340.009 ± 0.0051.75 ± 0.07175.00 ± 31.10
43.56 ± 0.060.265 ± 0.0032.20 ± 0.28253.50 ± 31.80
83.46 ± 0.010.314 ± 0.0412.20 ± 0.28228.50 ± 26.20
123.56 ± 0.060.276 ± 0.1101.40 ± 0.00203.50 ± 26.20
203.46 ± 0.000.332 ± 0.0151.80 ± 0.28208.00 ± 4.24
303.53 ± 0.010.317 ± 0.0051.70 ± 0.14208.00 ± 62.20
Table 2. Total Mesophilic Aerobic Bacteria (TMAB), Lactic Acid Bacteria Count (LAB) and yeast counts of fermented black carrot juice samples.
Table 2. Total Mesophilic Aerobic Bacteria (TMAB), Lactic Acid Bacteria Count (LAB) and yeast counts of fermented black carrot juice samples.
Ultrasound Time (min)Fermentation Period (Day)TMAB
(log cfu/mL)		LAB
(log cfu/mL)		Yeast
(log cfu/mL)
009.19 ± 0.06 abc7.81 ± 0.39 ab7.17 ± 0.01 ab
89.15 ± 0.01 abc8.37 ± 0.06 a6.03 ± 0.16 bcd
128.10 ± 0.23 de7.23 ± 0.26 bc4.23 ± 0.10 e
509.88 ± 0.06 a6.03 ± 0.12 d7.92 ± 0.05 a
89.10 ± 0.00 abc7.80 ± 0.22 ab5.60 ± 0.05 cd
127.15 ± 0.50 f7.05 ± 0.361 bcd4.83 ± 0.38 de
1509.76 ± 0.09 ab6.39 ± 0.05 cd8.14 ± 0.11 a
88.98 ± 0.07 bc8.50 ± 0.28 a6.53 ± 0.01 bc
127.41 ± 0.28 ef7.75 ± 0.52 ab5.05 ± 0.52 de
3009.62 ± 0.16 ab7.61 ± 0.15 ab8.15 ± 0.12 a
88.42 ± 0.19 cd8.32 ± 0.22 a6.27 ± 0.28 bc
127.70 ± 0.22 def8.02 ± 0.13 ab5.84 ± 0.56 cd
Different letters in the same column indicate statistically significant differences between samples.
Table 3. Color parameters of fermented black carrot juice samples.
Table 3. Color parameters of fermented black carrot juice samples.
Ultrasound Time (min)Fermentation Period (Day)L*a*b*C*h
0070.83 ± 1.940.78 ± 0.145.02 ± 0.625.08 ± 0.6381.16 ± 0.49
427.59 ± 3.2160.36 ± 2.3745.41 ± 4.2675.55 ± 4.4536.92 ± 1.51
822.72 ± 0.3056.06 ± 0.2438.15 ± 0.42467.81 ± 0.4434.24 ± 0.18
1223.42 ± 0.0056.46 ± 0.2739.34 ± 0.04268.82 ± 0.2134.87 ± 0.16
2024.74 ± 3.9057.75 ± 3.8541.56 ± 6.6371.19 ± 6.9835.62 ± 2.53
3020.10 ± 2.6252.48 ± 3.0233.68 ± 4.4362.38 ± 4.9432.61 ± 1.93
5068.30 ± 1.770.70 ± 0.154.11 ± 0.334.170 ± 0.3580.47 ± 1.31
427.63 ± 3.1660.73 ± 2.4345.69 ± 4.2676.02 ± 4.5036.91 ± 1.47
825.01 ± 2.6358.28 ± 2.3741.98 ± 4.3671.85 ± 4.4735.71 ± 1.73
1224.13 ± 3.6357.16 ± 3.2240.56 ± 6.1770.13 ± 6.1935.24 ± 2.60
2021.71 ± 0.7555.04 ± 0.8536.37 ± 1.3165.97 ± 1.4333.45 ± 0.54
3022.13 ± 4.3354.78 ± 4.6037.15 ± 7.4266.24 ± 7.9833.95 ± 3.10
15068.77 ± 0.110.83 ± 0.044.905 ± 0.5164.98 ± 0.5280.38 ± 0.53
426.27 ± 2.9259.41 ± 2.5043.94 ± 4.6773.91 ± 4.7936.42 ± 1.76
820.24 ± 0.8553.58 ± 0.9033.87 ± 1.3663.39 ± 1.4832.30 ± 0.60
1225.52 ± 0.1958.85 ± 0.1342.95 ± 0.2472.85 ± 0.2636.13 ± 0.09
2022.32 ± 5.9054.98 ± 6.5537.38 ± 10.1566.56 ± 11.1233.87 ± 4.10
3020.69 ± 6.1753.16 ± 7.1434.63 ± 10.6863.55 ± 11.8032.64 ± 4.64
30068.25 ± 3.760.91 ± 0.235.56 ± 0.725.64 ± 0.7680.83 ± 1.22
424.91 ± 1.8558.21 ± 1.7741.88 ± 3.1171.72 ± 3.2535.71 ± 1.20
823.23 ± 1.9056.62 ± 1.9838.99 ± 3.2168.75 ± 3.4534.52 ± 1.27
1225.01 ± 5.4757.91 ± 5.0141.70 ± 8.8871.42 ± 9.2535.53 ± 3.46
2023.27 ± 1.9256.44 ± 1.9239.05 ± 3.3868.65 ± 3.5034.65 ± 1.41
3024.41 ± 3.0656.83 ± 2.8640.86 ± 5.0969.51 ± 4.5835.64 ± 2.02
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Ercan, M.; Akbulut, M.; Çoklar, H.; Demirci, T. Impacts of Sonication on Fermentation Process and Physicochemical, Microbiological and Sensorial Characteristics of Fermented Black Carrot Juice. Fermentation 2025, 11, 475. https://doi.org/10.3390/fermentation11080475

AMA Style

Ercan M, Akbulut M, Çoklar H, Demirci T. Impacts of Sonication on Fermentation Process and Physicochemical, Microbiological and Sensorial Characteristics of Fermented Black Carrot Juice. Fermentation. 2025; 11(8):475. https://doi.org/10.3390/fermentation11080475

Chicago/Turabian Style

Ercan, Muhammet, Mehmet Akbulut, Hacer Çoklar, and Talha Demirci. 2025. "Impacts of Sonication on Fermentation Process and Physicochemical, Microbiological and Sensorial Characteristics of Fermented Black Carrot Juice" Fermentation 11, no. 8: 475. https://doi.org/10.3390/fermentation11080475

APA Style

Ercan, M., Akbulut, M., Çoklar, H., & Demirci, T. (2025). Impacts of Sonication on Fermentation Process and Physicochemical, Microbiological and Sensorial Characteristics of Fermented Black Carrot Juice. Fermentation, 11(8), 475. https://doi.org/10.3390/fermentation11080475

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