Traditional homemade probiotic drink “Kanji” versus marketed probiotic drink: Critical representation of hidden therapeutic potentials

Gagandeep Singh; Malika Arora; Mukesh Maithani; Moni Kumari; Parveen Bansal

doi:10.4103/jihs.jihs_31_20

ORIGINAL ARTICLE

Year : 2021 | Volume : 9 | Issue : 1 | Page : 13-18

Traditional homemade probiotic drink “Kanji” versus marketed probiotic drink: Critical representation of hidden therapeutic potentials

Gagandeep Singh¹, Malika Arora², Mukesh Maithani², Moni Kumari², Parveen Bansal¹
¹ University Centre of Excellence in Research, Baba Farid University of Health Sciences, Faridkot, Punjab, India
² Multi-Disciplinary Research Unit, Guru Gobind Singh Medical College and Hospital, Faridkot, Punjab, India

Date of Submission	13-Oct-2020
Date of Decision	15-Mar-2021
Date of Acceptance	25-Mar-2021
Date of Web Publication	17-Aug-2021

Correspondence Address:
Dr. Malika Arora
Multidisciplinary Research Unit, Guru Gobind Singh Medical College and Hospital, Baba Farid University of Health Sciences, Faridkot - 151 203, Punjab
India

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jihs.jihs_31_20

Abstract

Background: The traditional homemade Kanji (commonly known as Kanjika) is an Indian fermented drink, which is prepared from black carrots and has indigenous probiotic species. It is being used since long back due to its high nutritional/therapeutic value and availability of anthocyanin pigment. However, the use of traditional food drinks is decreasing and these are being replaced by marketed probiotic drinks. At present, probiotics are regulated as per their intended use, and there are certain confusions and challenges in regulatory guidelines. Hence, ensuring safe, swift, and successful usage of probiotics is the urgent need of the hour. Aims and objective: The present study was aimed to investigate homemade Kanji and a marketed formulation for its analytical/ biological studies in order to compare their therapeutic potency. Materials and Methods: A marketed probiotic drink was procured from the local market and Kanji was prepared at home using a traditional method. Results: The preliminary evaluation of kanji over marketed probiotic drink has shown that the homemade drink Kanji is having better probiotic, antioxidant, and antimicrobial profile in comparison to the marketed products. Conclusion: The results concluded that the traditional homemade drinks are more effective and safe beverages even as per today's lifestyle.

Keywords: Antimicrobial activity, antioxidant activity, Kanji, probiotics, traditional drinks

How to cite this article:
Singh G, Arora M, Maithani M, Kumari M, Bansal P. Traditional homemade probiotic drink “Kanji” versus marketed probiotic drink: Critical representation of hidden therapeutic potentials. J Integr Health Sci 2021;9:13-8

How to cite this URL:
Singh G, Arora M, Maithani M, Kumari M, Bansal P. Traditional homemade probiotic drink “Kanji” versus marketed probiotic drink: Critical representation of hidden therapeutic potentials. J Integr Health Sci [serial online] 2021 [cited 2023 Jun 4];9:13-8. Available from: https://www.jihs.in/text.asp?2021/9/1/13/323953

Introduction

In ancient times, our predecessors have exploited the power of microbes to make new foods though the science behind them was not known to them. The basic concept behind it was the fermentation. Fermentation usually alludes to the conversion of sugar to alcohol using yeast, whereas in some related processes, the use of bacteria such as Lactobacillus has been exploited in manufacturing of food products such as yogurt and sauerkraut.^[1] Indian culture is also rich in preparing and preserving foods using such traditional methods. Fermentation is one of the inexpensive and noticeable techniques used since prehistoric time for enhancing the taste and quality of food.^[2] Due to fermentation, there are a decrease in pH and an increase in titratable acid, which helps in the preservation of food by inhibiting the growth of contaminants.^[3] Although it is the earliest food preservation practice, it is still being accomplished at local or household level in India.^[4] There are various categories of fermented foods such as milk- and buttermilk-based fermented foods, cereal-based fermented foods, meat-based fermented foods, and vegetable- and fruit-based fermented foods.^[5]

Fermented Kanji (commonly known as Kanjika) is an Indian fermented drink, which is prepared from black carrots (Daucus carota) and is highly acidic in nature.^[6] It is being considered that the preparation is extremely beneficial for those who find it difficult to digest milk products. It is being prescribed by ayurvedic acharyas and medical practitioner since long back due to its high nutritional value, antioxidant activity, therapeutic potential, and availability of anthocyanin pigment.^[7] In addition, it has been observed that beetroot Kanji have cooling as well as soothing properties, which is found to be beneficial for preventing infection as well as other diseases.^[8] Moreover, it is a quite prevalent remedy for the treatment of gastritis, appetite loss, and liver disorders. It is prepared mainly in the early summer season and used comprehensively as an appetizer.^[9] Latif et al. reported the use of Kanji as a protective drink against oxidative stress.^[10] These beneficial properties may be due to availability of various indigenous probiotic microbes such as lactic acid bacteria (LAB) and nutritional components of carrot or beetroot.

Traditional foods have always stood upright as life saviors with potential therapeutic effects since ages. However, with the advancement of technology, the traditional foods faced exit from market followed by replacement with new so-called probiotic products. The therapeutic efficacy/potential of these formulations/products is under scanner. Till date, there is no study or evidence demonstrating its safe use as well as its safety/efficacy profile. Due to availability of catchy slogans and rosy picture of such products, the consumers are being attracted toward such products without knowing its consequences on human health. However, such claimed therapeutic effect does not exist in scientific literature. Recently, there is a resurgence of the use of traditional products of organic value. Hence, there is a need to establish the scientific evidences for homemade products that are accessible to consumers at very low price as compared to marketed product (MP). On the other hand, black carrots in India do not find much customer appreciation as a vegetable but are only utilized in rural areas for preparing fermented refreshment drink like Kanji.^[11] Despite the evident advantages of black carrots as a source of natural colorant, there is very less number of reports on its processing. Regardless of its regular consumption in India as an appetizer and soothing drink, it has not been explored much scientifically. In contrary part, we cannot trust on marketed probiotic products due to its inappropriate manufacturing and labeling in the absence of harmonized regulatory guidelines.^[12] There is absence of stringent harmonized guidelines for safe manufacturing of probiotic products.^[13] As a matter of fact, it is very difficult to rely upon the manufacturer's claimed therapeutic benefits, so with this intention, this study was conducted on a homemade product “Kanji” and its potentials have been compared with a known MP (name of formulation masked due to conflict of interest).

Materials and Methods

A marketed probiotic drink was procured from the local market and Kanji was prepared at home using a traditional method.^[14]

Preparation of Kanji

250 g black carrots (Daucus carota) were bought from the local vegetable market.

Preparation of carrots

The black carrots were properly washed with running water to remove dirt and impurities as well. Later, the carrots were peeled and chopped into tiny uniform pieces. The chopped carrots were transferred to a sterile container containing 1 l of RO filter water (water was premixed 30 g of sugar).

Addition of spices

A measured quantity, i.e., 500 mg of red chili powder and 500 mg of black chili powder, was added to mixture followed by addition of 10 g of mustard seed powder.

Aroma development

The mixture was set aside for 7–10 days in glass container for the development of aroma as well as taste. The Kanji prepared was evaluated as per following details.

pH evaluation

Acid is a major byproduct of fermentation and its presence can be checked by change in pH. The marketed probiotic drink and homemade Kanji were also subjected to change in pH. A periodic check was done on both the products to keep an eye on the pH of the products. For the same, pH of both the products was evaluated time to time on days 0, 7, 14, 21, and 28 using pH meter.

Microbial evaluation

The marketed probiotic drink and homemade Kanji were evaluated for microbial content using various selective and enrichment media. The bacterial colonies were isolated from both the abovementioned products using different media such as nutrient agar, De Man, Rogosa, and Sharpe agar (MRS agar), MacConkey agar (MA), and Sabouraud's dextrose agar (SDA). Respective serially diluted samples were inoculated onto these growth mediums in triplicates and were incubated at 37°C in aerobic conditions for 24 h, except MRS agar (anaerobic, 37°C, 48 h) and SDA plates (aerobic, 28°C, 48–96 h). The plates were used for determining the count of viable bacterial/fungal colonies, respectively, after their incubation.

Physiological and biochemical characterization

The colonies isolated from marketed probiotic drink and Kanji were identified by various standard morphological and biochemical examinations.^{[15],[16],[17],[18],[19],[20],[21]}

Antimicrobial activity

Some probiotic strains are known to be antagonistic for other pathogenic organisms, which might be due to production of some antimicrobial metabolites or enzymes.^[22] To check this activity of selected products, agar well diffusion assay was done against pathogenic strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Wells of 7 mm diameter were made on Mueller-Hinton agar plates, which were previously seeded with the abovementioned test organisms. 100 μl of cell-free extract of both the probiotic products was added into these wells and incubated at 37°C. After 24 h of incubation, diameter of zone of inhibition was measured by using a ruler. Inhibition zone of <10 mm, 10–20 mm, and more than 20 mm was considered as low, intermediate, and strong inhibition, respectively.^[23]

Antioxidant activity

Antioxidant activity of both the products was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The antioxidants are able to reduce the free, stable, and purple-colored DPPH radical to the colorless diphenyl-picryl-hydrazine.^[24] Compound with more antioxidant activity will be able to scavenge more DPPH radicals and make it more colorless. In this assay, 0.1 mM DPPH solution prepared in methanol was used to evaluate scavenging activity. 1 ml of methanolic DPPH solution was added to 1 ml of different concentrations (10 μg/ml, 50 μg/ml, and 100 μg/ml) of the standard (ascorbic acid) and both probiotic drinks. The standard and test solutions were kept in dark for 30 min for reaction, and absorbance was noted at 517 nm using ELISA microplate reader (Thermo Multiskan). Reduction in the absorbance of test solution as compared to control will indicate antioxidant activity. All the reactions were performed in triplicate, and scavenging activity of test solutions was determined by the following equation.^[25]

Acontrol is the absorbance value of DPPH without sample and Asample is the absorbance value of sample mixed with DPPH. Half-maximal inhibitory concentration (IC50) is a measure of the ability of the compound to inhibit a specific biochemical of biological function. It was calculated by plotting scatter plot chart and a standard reference line. The line of equation (y = mx + c) from bar graph was used to obtain the value of concentration (x) of compound required to scavenge the 50% (y = 50) of the DPPH radicals.

Statistical analysis

All the experiments were done in triplicates, and results were represented as mean ± standard deviation. One-way analysis of variance (ANOVA) test was done at α = 0.05 (level of significance), so as to determine the significant difference between the antioxidant properties of both products.

Results

pH analysis

PH was monitored from day 0 (day of preparation) to day 28. It was observed that on the day of preparation, pH of both the products was near to neutral. However, with due storage at room temperature, a huge decline in the pH was observed. In the case of Kanji, it dropped from 6.41 ± 0.07 to 3.47 ± 0.04 and from 6.20 ± 0.04 to 3.90 ± 0.29 for a marketed probiotic product. The comparative analysis of pH is depicted in [Figure 1]. This decline in pH can be due to fermentation of sugars present in products into lactic acid. Due to an increase in acidic content, similar change in pH was also recently reported by Lamba et al. in 2019.

Figure 1: Change in pH* of probiotic products due to storage (*Average of three determinations)

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Microbial evaluation

Both the marketed probiotic drink and the homemade Kanji were screened for the presence of various microbes such as coliforms, LAB, yeast, and mold by spreading them on different selective and enriched media. Total 11 isolates were obtained from both the products, out of which 7 (K1–K7) were isolated from Kanji and 4 (MP1–MP4) were from a marketed probiotic product. Various properties of these isolates such as colony morphology, cell characteristics, and biochemical properties are listed in [Table 1]. It has been observed that total plate count was 3.90 × 10⁶ CFU/ml and 1.43 × 10⁶ CFU/ml in Kanji and MP, respectively. Kanji had its indigenous bacteria which can be either probiotics or some other endophytes, and it has been hypothesized that due to which, a total load of bacteria in Kanji was reported to be more than a MP. MA was used for a coliform count, which was 2.70 × 10⁴ CFU/ml in Kanji and no colony was observed in case of MP.

Table 1: Morphological, physiological, as well as biochemical properties of different bacterial isolates from Kanji and marketed probiotic product (K1-K7 and MP1-MP4: Bacteria isolated from Kanji and marketed probiotic product, respectively)

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Coliform content in Kanji might be due to the use of nonsterilized water, or the carrots used for preparation might be irrigated with contaminated water. As the food industries maintain proper care regarding contamination by such organisms, no coliform load was reported in a MP, which can be due to the use of pasteurized raw material. Due to the storage of food products for a longer time, it may get contaminated with various molds. To check such content, both products were spread plated on SDA medium, it was observed that there was no growth in the case of Kanji, but 1.33 × 10⁵ CFU/ml was reported in MP. As the MP was not as fresh as Kanji, it showed the growth on SDA medium which might be due to the presence of various spores in the product. Enriched medium MRS agar was used for determining a load of probiotic bacteria in the products. LAB load in Kanji and MP was 3.30 × CFU/ml and 3.20 × 10⁶ CFU/ml, respectively. There is no significant difference between the LAB loads in both the products. It is very much clear that the naturally fermented probiotic products are more reliable and much safe to be used for any age group of people.

Antimicrobial activity

Antimicrobial activity of Kanji and marketed probiotic product was examined against three test organisms, i.e., E. coli, P. aeruginosa, and S. aureus, by using well diffusion assay. It was seen that homemade Kanji exhibited strong inhibition with zone of more than 20 mm. While very less inhibition activity was seen in case of MP only against S. aureus. Antimicrobial activity of probiotic strains isolated from curd was also reported in a study of Prabhurajeshwar et al. in 2017. This study also proved that this inhibitory action of probiotic strains is not due to organic acids, but there is a production of some bacteriocins too. Overall, Kanji has shown strong inhibition against pathogenic organisms in comparison to MP in [Table 2] and [Figure 2].

Table 2: Inhibitory action of probiotic products against pathogenic test organism

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Figure 2: Antimicrobial activity of Kanji and marketed product against Escherichia coli (a), Staphylococcus aureus (b), and Pseudomonas aeruginosa (c)

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Antioxidant activity

DPPH assay was done to determine the antioxidant activity of the probiotic products. It was observed that Kanji had a greater antioxidant property as compared to a marketed probiotic product. DPPH radical scavenging activity (%) of Kanji was 65.869 ± 0.06, 54.365 ± 0.08, and 31.393 ± 0.013 and that of MP was 34.413 ± 0.09, 27.235 ± 0.03, and 17.983 ± 0.005 at concentration of 100 μg/ml, 50 μg/ml, and 10 μg/ml, respectively [Table 3]. IC50 value comes out to be 52.78 μg/ml and 182.98 μg/ml in case of Kanji and MP, respectively.

Table 3: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (percentage) and half-maximal inhibitory concentration (μg/ml) at different concentrations of probiotic products

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It is quite clear from this study that Kanji has a bioactive potential to scavenge the reactive oxygen species and other radicals. This property may be due to the nutritional components of carrots or indigenous probiotic microorganisms. Hence, there is a need to explore the actual reason and isolate the compounds responsible for antioxidant potential of Kanji. High radical scavenging activity of Kanji is considered to be due to different types of pigment present in it. These pigments are in the form of flavonoids and carotenoids that impart color in addition to antioxidant properties.^[26] High antioxidant potential of fermented beverage prepared from carrot was also reported by Latif et al. in 2013. The difference between the antioxidant properties of both products is highly significant at 0.05 level of significance, as depicted by one-way ANOVA shown in [Table 4].

Table 4: Analysis of variance of percentage scavenging activity of Kanji and marketed probiotic product

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Conclusion

The use of probiotics not new to us as the base of probiotic products is the fermentation. The fermented products are being exploited since Vedic time due to their disease alternative properties. The present study was conducted to explore the anti-oxidation as well as antimicrobial potential of Kanji in comparison to commercially available fermented probiotic products. Nowadays, it has been observed that the peoples are relying more upon the probiotic products available in the market due to their rosy picture with respect to their therapeutic potential, but the present study highlighted that the homemade products are much more beneficial in comparison to MPs. Moreover, it has been observed that probiotic content was better in homemade Kanji. In addition, a highly significant difference between antioxidant and antimicrobial potentials of Kanji in contrast to MPs has been recorded. Hence, it has been concluded that our homemade recipes are still effective beneficial as well as safe in today's lifestyle. Furthermore, due to bioactive properties in homemade drinks, there is a great need to explore such products in detail, and this study signifies that there is an urgent need to highlight the benefits of traditional homemade drinks to general public for their awareness. However, such studies encourage the general public to consume traditional and natural food products which can deliver good nutrition as well as support in prevention of routine gastrointestinal troubles.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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