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Article

Development and Characterization of Cultured Buttermilk Fortified with Spirulina plantensis and Its Physico-Chemical and Functional Characteristics

by
Hency Rose
1,
Shiva Bakshi
1,
Prajasattak Kanetkar
1,
Smitha J. Lukose
2,
Jude Felix
1,
Satya Prakash Yadav
1,
Pankaj Kumar Gupta
1 and
Vinod Kumar Paswan
1,*
1
Department of Dairy Science and Food Technology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, India
2
Department of Dairy Chemistry, Verghese Kurien Institute of Dairy and Food Technology, Kerala Veterinary and Animal Sciences University, Mannuthy, Thrissur 680651, India
*
Author to whom correspondence should be addressed.
Dairy 2023, 4(2), 271-284; https://doi.org/10.3390/dairy4020019
Submission received: 28 February 2023 / Revised: 21 March 2023 / Accepted: 25 March 2023 / Published: 28 March 2023
(This article belongs to the Section Milk Processing)
Browse Figures
Review Reports Versions Notes

Abstract

:
In recent years, there has been an unprecedented increase in the demand for fermented dairy products due to medical recommendations and lifestyle preferences. Cultured buttermilk, as an ancient fermented dairy beverage, is an appropriate product choice in this context. This study presents a novel cultured buttermilk formulated by fortification with high protein microalgae Spirulina platensis, thus making it valuable and attractive because of its antioxidant properties. The fermentation process, nutraceutical properties, and sensory characteristics of developed cultured buttermilk with various concentrations of Spirulina (0.25, 0.5, and 1%) were compared with the control sample (0% Spirulina buttermilk). Different concentrations of Spirulina in buttermilk result in a significant increase in chlorophyll and carotenoid content, boosting its antioxidant properties. The study also evaluated the prebiotic properties of Spirulina, thus, demonstrating its ability to promote a healthy digestive system. It was found that the addition of 0.25% Spirulina was able to ferment the product more quickly and retained the sensory acceptability of the finished product. The protein content, free radical scavenging activity, chlorophyll, carotenoid, and total phenolic content of 0.25% Spirulina-fortified buttermilk was 1.83%, 48.19%, 30.9 mg/g, 8.24 mg/g, and 4.21 mg/g GAE, respectively. Based on the results obtained, it was concluded that cultured buttermilk with a high nutritional value and functional health benefits can be developed by fortification with 0.25% Spirulina as a natural ingredient.

1. Introduction

The nutritional and health benefits of traditional fermented dairy products drive growing consumer interest because they promote the intestinal bacterial microbiota thus promising healthy living and longer life expectancy [1,2]. Fermented milk products have been an integral part of Indian diets since ancient times. The products are the result of interactive processes that occur while surplus milk is being stored [3]. However, it was Louis Pasteur who first provided an explanation for these interactive processes through “fermentation theory” [4]. Fermented milk products have enhanced bioavailability, shelf life, and safety contributed by the bioactive metabolites produced by lactic acid bacteria (LAB). Fermented milk products possess desired and predictable sensory, rheological, nutritional, and functional changes, which have increased the economic value of fermented milk products.
When fermented milk or cream is churned into butter, buttermilk is produced as a byproduct, which is classified as a functional food, predominantly due to its emulsifying power contributed by the polar lipids and milk fat globule membrane (MFGM) components [5]. It is possible to describe buttermilk as traditional/conventional buttermilk or as cultured buttermilk [6]. Churning is used to separate the white butter from the curd, and the remaining liquid is known as cultured buttermilk. Curd manufacturing, curd homogenization, and water dilution are all steps in India’s commercial manufacture of cultured buttermilk [7]. Cultured buttermilk is a nutritious and health-promoting food beverage because it is higher in phospholipid content and an excellent source of calcium, phosphorus, vitamin B2, vitamin B12, pantothenic acid-vitamin B5, potassium, zinc, protein, iodine, and molybdenum. As a result of the consumption of cultured buttermilk in the diet, one can improve digestion, increase immunity, and reduce cholesterol levels in the blood [8]. The term “buttermilk” is ambiguous and can be confused with or related to a range of other products, including natural buttermilk, cultured buttermilk, sour milk, cultured milk, cultured skimmed milk, certain Scandinavian fermented milks, and Bulgarian fermented milks, depending on the region [9].
Cultured buttermilk is a well-known fermented dairy product with medicinal benefits, sometimes referred to as “Chhash” in South Asian nations [10]. Due to its pleasant taste and remarkable health benefits, buttermilk has become more and more popular among consumers and is a crucial component in many Ayurvedic compositions [11]. Churning cultured cream or cultured milk curd is the traditional method for making cultured buttermilk. Adding culture to low-fat milk, followed by fermentation and homogenization, is the current method for making cultured buttermilk. The commercial manufacture of dahi, or cultured buttermilk, in India uses a mixed strain of thermophilic and mesophilic homofermentative bacterial cultures.
Despite the high nutritional benefits, cultured buttermilk is naturally deficient in vitamin C, iron, and dietary fiber [12]. Therefore, it is essential to examine buttermilk’s qualities and supplement it. One of the best strategies to improve the overall dietary intake of food with the fewest negative consequences, according to nutritionists, is to fortify milk products using natural resources [13]. Protein-based natural additives are now replacing the use of synthetic food additives that may have neurological, gastrointestinal, and respiratory disturbances, and side effects [14,15]. Almost all natural additives come from plant derivatives and extracts. Throughout history, microalgae have been used as a natural additive. Over the past few decades, researchers have observed that algae can be used as a protein source. A variety of microalgae have been studied. However, Spirulina, namely S. platensis, has been studied more than others because of its rich components, beneficial effects, and nontoxic properties [16].
Spirulina platensis is a microalga that has high macro and micronutrient content that helps to fight hunger and malnutrition. Spirulina is a nutrient-dense food that can be used therapeutically, addressing a wide array of nutritional requirements. The Intergovernmental Institution for Micro-algae Spirulina Against Malnutrition (IIMSAM), an intergovernmental institution, has advocated Spirulina as a high-nutrient food since the mid-1970s for pregnant women and newborn infants [17]. Additionally, the National Aeronautics and Space Administration (NASA) and the European Space Agency (ESA) have both suggested Spirulina as the best and most sustainable space food (for long-term space missions) because of its concentrated macro- and micronutrient content [18]. Spirulina has demonstrated efficacy as a nutraceutical food for several ailments. As a dietary supplement, Spirulina has reportedly been shown to be effective in preventing and treating hypercholesterolemia, as well as managing allergies, cancer, toxicity, cardiovascular disease, and diabetes [19,20]. This is because of the chemical components of the food, which contains high levels of protein along with all nine required amino acids, vital fatty acids, minerals, pigment, and vitamins. In addition to providing nourishment, it has been discovered to have antioxidant, antibacterial, and antifungal properties against several human infections [21], and to encourage the formation of lactic acid bacteria in milk and dairy products [22]. Spirulina platensis has been used in a variety of products, including ayran, a fermented dairy beverage [23], Kefir [24], yoghurt [22,25,26,27,28], soy-yogurt biscuits [29], ice cream [30,31,32], cheese [33,34], protein bars [35], and others. To develop high-quality foods with a greater nutritional content, microalgae have been commercially exploited and used as functional components [36]. However, there have been no reports to date regarding fortification of buttermilk with Spirulina.
With this background, the current study was planned to produce spiced buttermilk fortified with Spirulina platensis to standardize the optimal concentration of Spirulina as a supplement in spiced buttermilk. Further, the effect of Spirulina on the physicochemical properties and antioxidative activity of the final product as well as its shelf life were assessed.

2. Materials and Methods

Pasteurized and standardized double-toned milk with 1.5% fat and 9.0% SNF was procured from Amul Milk outlet, Varanasi, India and stored at 4 °C until required. A freeze-dried direct vat-set (DVS) yoghurt culture (NCDC-167) containing a mixed strain of thermophilic and mesophilic homofermentative bacterial culture was obtained from the National Collection for Dairy Culture, Karnal, India. The culture was stored at −18 °C until used. Spirulina was procured from Heilen Biopharm Pvt. Ltd., India and was stored in a refrigerator until used. The nutritional composition, antioxidant activity, and total phenolic content of the procured Spirulina powder was evaluated adopting a standard protocol.

2.1. Preparation of Cultured Buttermilk

Preparation of probiotic buttermilk with Lactobacillus reuteri was optimized by Rodas et al. [37]. The same protocol was used for the development of fortified spiced buttermilk with minor modifications and by incorporation of Spirulina powder.
To produce the Spirulina-milk blend, Spirulina biomass (85–90%) was added to toned milk at a 2% (w/w) concentration at the room temperature (27 °C). The preliminary study indicated that at this rate (2% w/w), Spirulina was completely soluble. Direct addition of Spirulina after the production of buttermilk was found undesirable due to the insoluble particles of Spirulina in the sample. Therefore, a constant concentration of the Spirulina–milk blend was produced. Afterward, to ensure optimal mixing of the Spirulina and milk, the mixture was homogenized using a homogenizer at pressures of 100 kg/cm2 for the first stage and 200 kg/cm2 for the second stage. To obtain aliquots of 0%, 0.25%, 0.50%, and 1.00% (w/w) Spirulina biomass, this mixture was diluted using toned milk. Spices (0.4% w/w) were added to the control milk (0%) and the Spirulina-milk blend followed by pasteurization at 90 °C for 10 min in a water bath. The pasteurized samples (100 mL each) were put into polystyrene cups that were pre-sterilized and were allowed to cool to 4 °C. These aliquots received an inoculation of 0.5% (v/w) NCDC-167 stock cultures. All of the samples were carefully wrapped in aluminum foil, stored in low density polyethylene (LDPE) bags, and incubated for seven hours at 42 °C in a temperature-controlled incubator. The samples were observed hourly until they attained a pH of 4.6–4.7 and an acidity of 0.8–0.9%. Curd setting was observed after 7 h of complete fermentation. Then, agitation or breaking of curd was performed using a laboratory blender at a speed of 10,000 rpm for 90 s. As soon as the cultured buttermilk samples were prepared, they were stored under refrigeration at 6–8 °C. The formulated cultured buttermilk samples were referred to as; T0 (Control Buttermilk), T1 (0.25% Spirulina Buttermilk), T2 (0.5% Spirulina Buttermilk), and T3 (1% Spirulina Buttermilk). Post development, these samples were further analyzed adopting standard scientific methodology after refrigeration for 7, 14, 21, and 28 days.

2.2. Physico-Chemical Analysis

After calibrating the pH meter with calibrated pH buffer solutions (4.0, 7.0, and 10.0), the pH of the unfortified and Spirulina-fortified cultured buttermilk samples was determined. Titratable Acidity (TA) was determined and calculated as a percentage of the lactic acid that was produced. The total solids of samples were determined using an oven in which the dry samples were kept at 105 °C overnight till a constant weight was attained. Dry samples were ignited in a muffle furnace at 550 °C to burn all the organic components and measure the amount of ash present. The protein level was determined by the Kjeldhal method using a nitrogen conversion factor of 6.38. The Gerber method was used to determine the sample’s fat content [38]. For determination of calcium, the method was adopted from Sehgal [39]. The total carbohydrates and energy (Kcal/100 mL) in the sample were calculated using the procedure of Cunniff [40]. All the analyses were performed in triplicate.

2.3. Antioxidant Analysis

2.3.1. DPPH Inhibition Activity

The method described by Kang and Saltveit [41] was used to measure the free radical scavenging activity of samples using the indicator 1,1-diphenyl-2-picrylhydrazil (DPPH). The DPPH radical scavenging activity was evaluated at 517 nm, and the results were presented as percentage of inhibition. All reactions were carried out in triplicates.

2.3.2. Analysis of Total Phenolic Content

The total phenolic content was determined by the Folin–Ciocalteu method [42] with some modifications. A total of 0.5 mL of the sample extract was taken in a test tube and 2.5 mL of diluted Folin–Ciocalteu (FC) Reagent was added followed by 1 mL of 7.5% Na2CO3. The test tube was covered with aluminum foil and incubated in the dark at room temperature for 1 h. Absorbance was measured at 760 nm in UV-1800 spectrophotometer. Gallic acid (0–800 mg/L) was used to produce standard calibration curve. The total phenolic content was expressed in mg of Gallic acid equivalents (GAE)/100 g. The standard curve with regression equation (y = 0.0769x + 0.0041, R2 = 0.998) was obtained using the absorbance against the Gallic acid concentration (mg/100 mg).

2.3.3. Analysis of Pigments

The chlorophyll and carotenoid content in the samples was determined as per Kumar et al. [43] and Lichtenthaler and Wellburn [44].
A total of 1 mL of each sample was centrifuged at 5000 rpm for 10 min. The pellet was dissolved in 1 mL of ethanol and then sonicated at 65 °C for 30 min. After sonication, the solutions were centrifuged at 10,000 rpm for 5 min. The pigment content was estimated by measuring the supernatant absorbance (A) at 666, 653, and 470 nm and calculated using the following equations:
Chlorophyll   a   ( mg L ) = 15.65 × A 666   − 7.340 × A 653
Chlorophyll   b ( mg L ) = 27.05 × A 653   − 11.21 × A 666
Total   Chlorophylls   ( mg L ) = Chlorophyll   a + Chlorophyll   b
Carotenoids   ( mg L )   =     ( 1000 × A 470     − 2.860 × [ Chlorophyll a ] − 85.9 × [ Chlorophyll b ] ) / 245  
Determination of C-Phycocyanin content was executed as per Pan-utai and Iamtham [45]. A total of 1 mL of each sample was centrifuged at 5000 rpm for 10 min. The pellet was dissolved in 1 mL of distilled water and then sonicated for 30 min. The supernatant was collected by centrifugation at 10,000 rpm for 5 min and measured for optical density at 615 and 652 nm. C-Phycocyanin content was calculated using the following equation. For purity, a ratio of A620/A280 was used.
C − phycocyanin   ( mg mL ) = ( O D 615 − 0.474 × O D 652 ) / 5.34

2.4. Sensory Evaluation

Sensory evaluation of the control and Spirulina-fortified cultured buttermilk samples was carried out using a 9-point hedonic scale of Like extremely (score-9), Like very much (score-8), Like moderately (score-7), Like slightly (score-6), Neither like nor dislike (Score-5), Dislike slightly (score-4), Dislike moderately (score-3), Dislike very much (score-2), Dislike extremely (Score-1). Twenty trained panelists were selected based on their previous experience and knowledge of sensory evaluation of dairy and milk-based products particularly those who were regular consumers of cultured buttermilk. Color and appearance, body and mouthfeel, flavor, and overall acceptability were the sensory parameters evaluated by the sensory panelists. All cultured buttermilk samples were taken out of the refrigerator and the serving temperature for samples ranged from 10 to 12 °C. Each cultured buttermilk sample was presented in a 100 g plastic cup filled with 50 g cultured buttermilk sample and labeled with a 3-digit code. Order of presentation of samples was randomized. Evaluation was divided into 3 sections: visual characteristics (color and appearance), texture (body and mouthfeel), and flavor evaluations. For visual attributes, the surface of each sample was examined in terms of any variation in color or unevenness. The visual analysis of cultured buttermilk was followed by texture and flavor evaluations.

2.5. Enumeration of Microorganisms

The samples comprising 1 mL of each were aseptically removed and serially diluted with 0.9% normal saline. For the enumeration of Streptococcus thermophilus and Lactobacillus bulgaricus M-17 agar and De Man, Rogosa and Sharpe (MRS) agar were used, respectively. The plates were incubated for 48 h at 37 °C [46]. Coliforms, yeast, and mold count were performed using the acidified Potato Dextrose Agar (PDA) and Violet Red Bile Agar (VRB), respectively, in accordance with the Standard Procedures for the Analysis of Milk Products [47]. The data were expressed as log CFU/mL. During the storage period, a microbial study was conducted using the spread plate method on the 1st, 3rd, 6th, 9th, and 12th days.

2.6. Statistical Analysis

All analytical determinations were performed at least in triplicate and values were expressed as the mean ± standard deviation (SD). One-way ANOVA and Duncan’s multiple comparison tests were used to compare results with significant differences (p < 0.05). IBM SPSS statistics software version 19 (IBM Corp., Armonk, NY, USA) was used to perform all statistical analyses.

3. Results and Discussion

3.1. Composition of Spirulina Powder

The Spirulina powder used in the study was procured from Heilen Biopharm Pvt. Ltd., Gandhinagar, India. The nutritional composition and antioxidant activities of the Spirulina powder is presented in Table 1. The procured Spirulina powder has 5.12% moisture, 59.27% protein, and 7.02% total ash content. The protein content and ash content of Spirulina as reported by earlier researchers ranged from 53 to 65% and 7.9–8.4%, respectively, when expressed on dry matter basis [48,49,50]. Thus, the protein and total ash contents of the procured Spirulina powder in the present study was within the range of reported values. Further, it was reported that Spirulina extract was able to inhibit 42–50% of free radicals, containing Total Phenolic Content (TPC) 4.51–26.64 mg GAE/g samples on dry matter basis [48,51]. The variable % DPPH inhibition (89.00 ± 1.02%) and total phenolic content (11.26 ± 0.04 mg GAE/100 g) in the procured Spirulina microalga may be due to different stage of maturity and harvest and processing such as grinding and drying conditions.

3.2. Preparation of Cultured Buttermilk

Figure 1 shows the pH and acidity levels of the different treatments measured over the course of the fermentation (4 hours). During the development of cultured buttermilk, caseinate particles entirely precipitate over a pH range of 4.6–4.7 [52]. Therefore, the duration of fermentation in the current study was determined by the time it took for the pH to reach 4.6–4.7. Due to the strong buffering properties of milk and the rapid bacterial growth during fermentation, increasing rates of titratable acidity and decreasing rates of pH were observed across all treatments [22]. The pH of the Spirulina-fortified buttermilk rapidly drops after 2 h, whereas the T0 takes a substantially longer period of 3 h. For instance, after 1–2 h of fermentation, the T3 sample showed a pH drop from 6.42 to 4.29 (a 2.27 reduction), whereas the pH of T0 declined from 6.42 to 6.2. Spirulina platensis was added to buttermilk samples, and as a result, the time it took to achieve pH 4.5 was considerably shortened (p < 0.05). The samples containing 0.25, 0.5, and 1% Spirulina powder did not significantly differ from one another, though. Additionally, these Spirulina-fortified treatments had a much faster increase in acidity (p < 0.05). It was found that there was a moderate agreement between the results of this study and those of earlier workers [22,25,53,54,55,56]. Spirulina-inoculated milk with S. thermophilus and L. bulgaricus decreased pH levels more rapidly than control samples during fermentation, resulting in a shorter incubation period.
As shown in Figure 2, each product was evaluated for color, flavor, mouthfeel, and overall acceptability. According to the hedonic scale, 82% of the trained sensory panelist indicated a positive response to T0 and 78% to T1. The majority of panelists underrated T3 with a score of 5 on the hedonic scale. Supplemental Spirulina creates compounds from the oxidation of minerals and lipids that not only function as pro-oxidant molecules but may also produce metallic off-flavors, resulting in an unfavorable flavor [57]. The sample T1 was not significantly different from T0 (Figure 2). When panelists were exposed to buttermilk with a 1% microalgae concentration (T3), it was found to have an unsuitable sensory property (p = 0.001). In addition, insoluble Spirulina particles produced a graininess mostly in T2 and T3, and a significant change in oral texture was observed (p < 0.001). Thus, it showed that the higher concentration of Spirulina decreases the solubility as compared to other samples, therefore higher concentrations were not accepted. The samples of T1 scored the highest for taste, texture, and mouthfeel control, while treatments containing T2 and T3 scored the lowest (p < 0.001). Thus, the highest sensory score and overall acceptability was obtained for T1 with 0.25% Spirulina-fortified buttermilk. Therefore, the buttermilk supplemented with 0.25% of Spirulina (T1) was selected as the best final product and was evaluated for further analysis.

3.3. Physicochemical Analysis

Buttermilk that was enriched with Spirulina (T1) and the control sample (T0) had comparable chemical compositions for most of the attributes. Nonetheless, Spirulina-treated buttermilk was shown to have more total solids (TS), protein, fat, and ash concentrations (Table 2). Both the control and the Spirulina-fortified buttermilk had total solids of 9.73% and 9.87%, respectively. Between these two samples, there was a significant difference in TS (p < 0.001). The fat content of the control (0.70) and fortified buttermilk sample (0.73) did not differ significantly (p < 0.05). However, the Spirulina-fortified buttermilk had a higher total protein value of 1.83 compared to the control sample (1.71) (p < 0.001). This was most likely caused by the high protein content of Spirulina [58]. The sample treated with Spirulina had the highest calcium concentration (82.05 mg/100 g on dry weight basis) (p < 0.001). This finding is in line with the findings of Ustun-Aytekin et al. [59], who reported that the calcium content of milk appeared to fall noticeably after fermentation, whereas Spirulina-enriched kefir was found to have higher levels of calcium than that of traditional kefir. Due to this high content of calcium, T1 had the higher ash content. The ash content found in T1 was 0.76%, which was significantly higher than the ash percentage of T0 (0.70%) (p < 0.05). When compared to T0, the energy value (Kcal/100 mL) reveals a significant difference (p < 0.001), with the highest value for T1 (40.29). This is due to the presence of the high carbohydrate content present in T1 (6.59). Changes in these factors, particularly in the amounts of protein, fat, and dietary fiber, may have an impact on other physicochemical characteristics including pH and titratable acidity.
The viscosity of a fluid describes its resistance to flow. It contributes directly to the mouthfeel characteristic of cultured buttermilk up to a certain degree. The presence of high viscosity in cultured buttermilk can also cause low phase separation since it reduces the likelihood of whey separation from the network when the viscosity of the buttermilk is high. The binding abilities or stabilizing between the serum and aqueous phase of Spirulina powder may have contributed to the higher viscosity (21.9 cP) values exhibited even in low concentrations of Spirulina powder added into cultured buttermilk. These results are similar with the findings of Mudgil et al. [7].
The pH and titratable acidity of T0 and T1 samples during the period of 12 days of storage at 4 °C are shown in Figure 2. In the current investigation, the pH of the treated sample dropped (from 4.60 to 4.51) during storage, indicating that lactic acidity (0.22 to 0.35%) developed throughout the storage period. This finding is in agreement with the data of earlier researchers [25,60,61]; they reported that Spirulina-added yoghurt had lower pH values than control samples during the storage period.

3.4. Antioxidant Properties

A great deal of recent attention has been paid to the potential role of enriched foods that contain bioactive compounds, generally obtained from microalgae, in the treatment of many human diseases. Microalgae are content with high amounts of free radical scavengers, which explains their high antioxidant activity. Hence, chlorophyll, carotenoid contents, total phenolic content, and DPPH-radical scavenging activity were measured (Table 3) to evaluate its antioxidant capacity.

3.4.1. DPPH Radical Scavenging Activity

As shown in Table 3, the experimental samples exhibited high antioxidant properties in terms of DPPH radical scavenging activity. The treatment sample T1 exhibited significantly higher (48.19% versus 41.99%) (p < 0.001) radical scavenging activity. It has been reported that higher antioxidant activity is observed when a higher ratio of microalga is used [62]. Studies with yogurts fortified with Chlorella vulgaris and Dunaliella sp. have also shown the microalgae-enhanced antioxidant activity of the yogurt. Incorporating Spirulina powders into the diet may increase the levels of chlorophylls [63], carotenoids [64], and phycocyanin [65], resulting in an increase in free radical scavenging.

3.4.2. Chlorophyll Content

Spirulina incorporation significantly improved the chlorophyll content from 0.33 mg/g in T0 to 30.95 mg/g in T1 (p < 0.001) besides the enhancement of DPPH inhibition activities (p < 0.05). This result was found to be similar to findings of Barkallah et al. [25]; they investigated the effects of Spirulina-fortified yoghurt and reported that the chlorophyll content of Spirulina-fortified yoghurt was 27.6 mg/g on a dry weight basis as compared to 0.25% in the control yoghurt. According to Ismaiel et al. [63], an increase in free radical scavenging can be attributed to an increase in chlorophyll content. Marzorati et al. described a study of chlorophyll extraction methods using supercritical CO2 and water [64]. They reported that the total chlorophyll content present in dried Spirulina powder was 9.1 mg/g. In the present investigation, the value obtained was much higher and it is inferred that the yield of chlorophyll content may vary depending on the different extraction processes.

3.4.3. Carotenoid Content

Carotenoids are a class of pigments found in nature. Spirulina contains up to 4000 mg/kg of carotenoids, with β-carotene dominating [65]. Carotenoids contain antioxidant, anti-aging, and anti-inflammatory properties, among other things. Carotenoids can quench singlet oxygen and scavenge radicals, resulting in the termination of oxidation chain reactions [66]. Because of their antioxidant characteristics, carotenoids are commonly utilized to treat cardiovascular diseases [67]. Barkallah et al. reported that the carotenoid content in T1 was 10.86 mg/g on a dry weight basis in their study [25]. Szmejda et al. [68] analyzed antioxidant compounds in ice cream supplemented with Spirulina extract. There was a significant difference (p < 0.001) in the carotenoid level present in the control (T0) and Spirulina extract-supplemented ice cream (T1) which were 0.072 mg/g and 8.24 mg/g, respectively. The study of Szmejda et al. indicated that when compared to non-supplemented flavor variants, the samples enhanced with Spirulina extract have a 3–3.5 times larger magnitude of carotenoids. Marzorati et al. studied the supercritical CO2 and water extraction methods for carotenoids, and they reported that total carotenoid content present in dried Spirulina powder was equal to 3.5 mg/g [66]. The yield of carotenoid content may also vary depending on the different extraction processes. The carotenoid content present in Spirulina-fortified buttermilk acts as a health-benefiting beverage at considerably low cost. Carotenoid as a color pigment also contributes to the antioxidant compound. However, the pigment quality present in buttermilk decreases upon storage.

3.4.4. C Phycocyanin Content

The C-Phycocyanin level was observed in T1 with 0.028 mg/g showing a significant difference (p < 0.001). This compound was absent in the control buttermilk. A similar result was observed in the study of Barkallah et al. on the effect of Spirulina-fortified yoghurt. They reported that the C-Phycocyanin content in T1 was 0.297 mg/g on a dry weight basis [25]. Marzorati et al. reported the yield of C-Phycocyanin using the water extraction method including electrocoagulation, dialysis, and protein salting-out method yielded- 250 mg/g of C-Phycocyanin with a very high purity of 2.2 [66]. Khandual et al. described different methods for extracting phycocyanin from Spirulina biomass (dry, wet, and frozen) [69]. They reported a greater phycocyanin content with water as the solvent, 46.65–54.65 mg/g biomass. The phycocyanin concentration of the phosphate buffer at pH 6 was 35.54–37.88 mg/g, while that of the phosphate buffer at pH 7 was 43.13–45.02 mg/g. When we compared different procedures, ultrasonication produced the highest phycocyanin content, 54.65 mg/g with water as the solvent, and the lowest, 35.54 mg/g with phosphate buffer at pH 6. It was observed that ultrasonication retrieved the most phycocyanin, followed by freeze/thaw, and sonication.

Purity of C-Phycocyanin

C-Phycocyanin with a purity of 0.7 is regarded as food grade and between 0.7 to 3.9 is called reactive grade, and greater than 3.9 is termed analytical grade [70]. However, the purity of the experimental sample T1 was 1.32%, which is above 0.7%. This shows that our results were included within the food grade range.

3.4.5. Total Phenolic Content (TPC)

A significant difference was observed between T0 and T1 for TPC (p < 0.001). As a result of the addition of Spirulina platensis to the cultured buttermilk, the total phenolic content was greatly increased in T1. A similar response was observed when the microalgae was added to gluten-free pasta samples [64]. In addition, Niccolai et al. identified an increase in phenol content as a result of fermentation of Spirulina biomass [71]. Previously, Liu et al. reported that Spirulina platensis is high in phenolic content (19.47 mg/g GAE) and fermentation enhances its effects [72].

3.5. Enumeration of Microorganism

The changes observed in the cultures of L. bulgaricus and S. thermophilus throughout the period of 15 days of storage at 4 °C are shown in Figure 3. After the fermentation process, the L. bulgaricus and S. thermophilus counts of the buttermilk samples were analyzed, and the initial counts of the microorganisms were found to be 9.12 log CFU/g and 9.13 log CFU/g, respectively. The results showed that Spirulina platensis had a significant impact on the lactobacilli and streptococci counts of treated samples compared to control sample milk throughout 12 days of cold storage (p < 0.05). Comparable results were reported by Güldas and Irkin [73] and Sudheendra [74]. Similarly, Beheshtipour et al. [22] found that lactic acid bacteria were more likely to survive in the presence of Spirulina platensis. According to Alizadeh Khaledabad et al. [60], probiotic populations of yoghurt samples were boosted by an increase in Spirulina platensis during its shelf-life. Furthermore, Spirulina platensis is known to contain further growth-inducing components, such as adenine, free amino acids, and linoleic acid [75,76,77]. This indicates that Spirulina has some prebiotic efficiencies that could help the growth of the culture. Varga and co-workers [55] were the first to identify this symbiotic relationship between Spirulina platensis and lactobacilli. Additionally, Gupta et al. [77] investigated how blue green algae affected probiotic bacteria. Due to the prebiotic properties of Spirulina, the product in our study may be regarded as a symbiotic product, although further evaluations and studies need to be conducted to determine Spirulina’s prebiotic efficiency.
None of the samples had any evidence of mold, yeast, or coliform bacteria during storage. Even after 12 days of storage at 4 °C, the lack of these bacteria demonstrated that the buttermilk were secure and uncontaminated. These findings imply that buttermilk processing took place in clean and hygienic conditions.

4. Conclusions

As an innovative and attractive ingredient, Spirulina powder can be successfully incorporated into spiced buttermilk as a means of enhancing the nutritional and physico-chemical properties without significantly altering its sensory acceptability. The study indicated that fortifying buttermilk with 0.25% Spirulina enhanced nutritional and functional properties by enhancing its antioxidant activity. The microbial evidence of this investigation suggest that Spirulina platensis may have the ability to encourage the growth of lactic acid bacteria and can be the effective prebiotic algal source. Thus, it can be used as a high-quality dietary supplement to enhance the nutritive and bioactive properties of food and dairy products. Further research is required to overcome its unpleasant flavor by modern encapsulating techniques and the efficient extraction of bioactive components such carotenoids and C-Phycocyanin using improved extraction techniques.

Author Contributions

Conceptualization, H.R. and V.K.P.; methodology, H.R., P.K. and J.F.; software, S.J.L.; validation, S.B., S.P.Y. and P.K.G.; formal analysis, H.R. investigation, J.F.; resources, V.K.P.; data curation, H.R., V.K.P. and S.J.L.; writing—original draft preparation, H.R., S.P.Y. and S.B.; writing—review and editing, P.K.G.; visualization, P.K.; supervision, V.K.P. and S.J.L. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Data Availability Statement

The original contributions presented in the study are included in the article; further inquiries can be directed to the corresponding author/s.

Acknowledgments

The corresponding author acknowledges the IoE Scheme research grant provided by Banaras Hindu University, Varanasi, India.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Bourrie, B.C.T.; Willing, B.P.; Cotter, P.D. The microbiota and health promoting characteristics of the fermented beverage kefir. Front. Microbiol. 2016, 7, 647. [Google Scholar] [CrossRef] [Green Version]
  2. Chen, M.; Ye, X.; Shen, D.; Ma, C. Modulatory effects of gut microbiota on constipation: The commercial beverage yakult shapes stool consistency. J. Neurogastroenterol. Motil. 2019, 25, 475–477. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  3. Tamang, J.P.; Cotter, P.D.; Endo, A.; Han, N.S.; Kort, R.; Liu, S.Q.; Mayo, B.; Westerik, N.; Hutkins, R. Fermented Foods in a Global Age: East meets West. Compr. Rev. Food Sci. Food Saf. 2020, 19, 184–217. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Barnett, J.A. A history of research on yeasts 2: Louis Pasteur and his contemporaries, 1850–1880. Yeast 2000, 16, 755–771. [Google Scholar] [CrossRef]
  5. Rombaut, R.; Van Camp, J.; Dewettinck, K. Phospho- and sphingolipid distribution during processing of milk, butter and whey. Int. J. Food Sci. Technol. 2006, 41, 435–443. [Google Scholar] [CrossRef]
  6. Gebreselassie, N.; Abrahamsen, R.K.; Beyene, F.; Abay, F.; Narvhus, J.A. Chemical composition of naturally fermented buttermilk. Int. J. Dairy Technol. 2016, 69, 200–208. [Google Scholar] [CrossRef]
  7. Mudgil, D.; Barak, S.; Darji, P. Development and characterization of functional cultured buttermilk utilizing Aloe vera juice. Food Biosci. 2016, 15, 105–109. [Google Scholar] [CrossRef]
  8. Nirgude, R.; Binorkar, S.V.; Parlikar, G.R.; Kirte, M.C.; Savant, D.P. Therapeutic and nutritional values of takra (buttermilk). Int. Res. J. Pharm. 2013, 4, 29–31. [Google Scholar]
  9. O’Connell, J.; Fox, P. Heat Stability of Buttermilk. J. Dairy Sci. 2000, 83, 1728–1732. [Google Scholar] [CrossRef]
  10. Ghanshyambhai, M.R.; Balakrishnan, S.; Aparnathi, K.D. Standardization of the method for utilization of paneer whey in cultured buttermilk. J. Food Sci. Technol. 2015, 52, 2788–2796. [Google Scholar] [CrossRef] [Green Version]
  11. Devi, M.C.A. Role of milk & milk products in traditional medicinal systems. Indian Dairym. 2010, 62, 114–119. [Google Scholar]
  12. Naidu, K.A. Vitamin C in human health and disease is still a mystery? An overview. Nutr. J. 2003, 2, 7. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Gahruie, H.H.; Eskandari, M.H.; Mesbahi, G.; Hanifpour, M.A. Scientific and technical aspects of yogurt fortification: A review. Food Sci. Hum. Wellness 2015, 4, 1–8. [Google Scholar] [CrossRef] [Green Version]
  14. Carocho, M.; Barreiro, M.F.; Morales, P.; Ferreira, I.C. Adding molecules to food, pros and cons: A review on synthetic and natural food additives. Compr. Rev. Food Sci. Food Saf. 2014, 13, 377–399. [Google Scholar] [CrossRef]
  15. Randhawa, S.; Bahna, S.L. Hypersensitivity reactions to food additives. Curr. Opin. Allergy Clin. Immunol. 2009, 9, 278–283. [Google Scholar] [CrossRef]
  16. Seyidoglu, N.; Inan, S.; Aydin, C. A prominent superfood: Spirulina platensis. Superfood and functional food the development of superfoods and their roles as medicine. Biology 2017, 22, 1–27. [Google Scholar] [CrossRef] [Green Version]
  17. Cai, J.; Lovatelli, A.; Aguilar-Manjarrez, J.; Cornish, L.; Dabbadie, L.; Desrochers, A.; Diffey, S.; Garrido Gamarro, E.; Geehan, J.; Hurtado, A.; et al. Seaweeds and Microalgae: An Overview for Unlocking Their Potential in Global Aquaculture Development; FAO Fisheries and Aquaculture Circular: Rome, Italy, 2021; p. 1229. [Google Scholar] [CrossRef]
  18. Satyaraj, E.; Reynolds, A.; Engler, R.; Labuda, J.; Sun, P. Supplementation of diets with spirulina influences immune and gut function in dogs. Front. Nutr. 2021, 8, 667072. [Google Scholar] [CrossRef]
  19. Karkos, P.D.; Leong, S.C.; Sivaji, N.; Assimakopoulos, D.A. Spirulina in clinical practice: Evidence-based human applications. Evid.-Based Complement. Altern. Med. 2011, 2011, 531053. [Google Scholar] [CrossRef] [Green Version]
  20. Thajuddin, N.; Subramanian, G. Cyanobacterial biodiversity and potential applications in biotechnology. Curr. Sci. 2005, 89, 47–57. [Google Scholar]
  21. Usharani, G.; Srinivasan, G.; Sivasakthi, S.; Saranraj, P. Antimicrobial activity of Spirulina platensis solvent extracts against pathogenic bacteria and fungi. Adv. Biol. Res. 2015, 9, 292–298. [Google Scholar]
  22. Beheshtipour, H.; Mortazavian, A.M.; Haratian, P.; Khosravi-Darani, K. Effects of Chlorella vulgaris and Arthrospira platensis addition on viability of probiotic bacteria in yogurt and its biochemical properties. Eur. Food Res. Technol. 2012, 235, 719–728, Erratum in Eur. Food Res. Technol. 2012, 235, 1213. [Google Scholar] [CrossRef]
  23. Çelekli, A.; Alslibi, Z.A.; Üseyin Bozkurt, H. Influence of incorporated Spirulina platensis on the growth of microflora and physicochemical properties of ayran as a functional food. Algal Res. 2019, 44, 101710. [Google Scholar] [CrossRef]
  24. Atik, D.S.; Gürbüz, B.; Bölük, E.; Palabıyık, I. Development of vegan kefir fortified with Spirulina platensis. Food Biosci. 2021, 42, 101050. [Google Scholar] [CrossRef]
  25. Barkallah, M.; Dammak, M.; Louati, I.; Hentati, F.; Hadrich, B.; Mechichi, T.; Ayadi, M.A.; Fendri, I.; Attia, H.; Abdelkafi, S. Effect of Spirulina platensis fortification on physicochemical, textural, antioxidant and sensory properties of yogurt during fermentation and storage. LWT 2017, 84, 323–330. [Google Scholar] [CrossRef]
  26. Debbabi, H.; Boubaker, B.; Gmati, T.; Chouaibi, M.; Boubaker, A.; Snoussi, A. Yogurt enrichment with Spirulina (Arthrospira platensis): Effect of storage on physicochemical parameters. In Recent Advances in Environmental Science from the Euro-Mediterranean and Surrounding Regions: Proceedings of the Euro-Mediterranean Conference for Environmental Integration (EMCEI-1), Sousse, Tunisia, 22–25 November 2017; pp. 1267–1268; Springer International Publishing: Berlin/Heidelberg, Germany, 2018. [Google Scholar] [CrossRef]
  27. Arslan, R.; Aksay, S. Investigation of sensorial and physicochemical properties of yoghurt colored with phycocyanin of Spirulina platensis. J. Food Process. Preserv. 2022, 46, e15941. [Google Scholar] [CrossRef]
  28. Agustini, T.W.; Soetrisnanto, D.; Ma’ruf, W.F. Study on chemical, physical, microbiological and sensory of yoghurt enriched by Spirulina platensis. Int. Food Res. J. 2017, 24, 367–371. [Google Scholar]
  29. Sengupta, S.; Koley, H.; Dutta, S.; Bhowal, J. Hypocholesterolemic effect of Spirulina platensis (SP) fortified functional soy yogurts on diet-induced hypercholesterolemia. J. Funct. Foods 2018, 48, 54–64. [Google Scholar] [CrossRef]
  30. Tiepo, C.B.V.; Gottardo, F.M.; Mortari, L.M.; Bertol, C.D.; Reinehr, C.O.; Colla, L.M. Addition of Spirulina platensis in handmade ice cream: Phisicochemical and sensory effects/Adição de Spirulina platensis em sorvete caseiro: Efeitos físico-químicos e sensoriais. Braz. J. Dev. 2021, 7, 88106–88123. [Google Scholar] [CrossRef]
  31. Rasouli, F.; Berenji, S.; Shahab Lavasani, A. Optimization of traditional Iranian ice cream formulation enriched with spirulina using response surface methodology. J. Food Technol. Nutr. 2017, 14, 15–28. [Google Scholar]
  32. Malik, P.; Kempanna, C.; Paul, A. Quality characteristics of ice cream enriched with Spirulina powder. Int. J. Food Nutr. Sci. 2013, 2, 44–50. [Google Scholar]
  33. Bosnea, L.; Terpou, A.; Pappa, E.; Kondyli, E.; Mataragas, M.; Markou, G.; Katsaros, G. Incorporation of Spirulina platensis on traditional greek soft cheese with respect to its nutritional and sensory perspectives. In Proceedings of the 1st International Electronic Conference on Food Science and Functional Foods, Online, 10–25 November 2020; Volume 70, p. 99. [Google Scholar]
  34. Terpou, A.; Bosnea, L.; Mataragkas, M.; Markou, G. Influence of Incorporated Arthrospira (Spirulina) platensis on the Growth of Microflora and Physicochemical Properties of Feta-Type Cheese as Functional Food. In Proceedings of the 1st International Electronic Conference on Food Science and Functional Foods, Online, 10–25 November 2020; Volume 70, p. 97. [Google Scholar]
  35. Kumar, A.; Mohanty, V.; Yashaswini, P. Development of high protein nutrition bar enriched with Spirulina plantensis for undernourished children. Curr. Res. Nutr. Food Sci. J. 2018, 6, 835–844. [Google Scholar] [CrossRef] [Green Version]
  36. Kreitlow, S.; Mundt, S.; Lindequist, U. Cyanobacteria—A potential source of new biologically active substances. In Progress in Industrial Microbiology; Elsevier: Amsterdam, The Netherlands, 1999; Volume 35, pp. 61–63. [Google Scholar] [CrossRef]
  37. Rodas, B.A.; Angulo, J.O.; De la Cruz, J.; Garcia, H.S. Preparation of probiotic buttermilk with Lactobacillus reuteri. Milchwissenschaft 2002, 57, 26. [Google Scholar]
  38. IS: 1224, (Part-I); Indian Standards. Determination of Fat by Gerber Method. Indian Standards Institution: New Delhi, India, 1981.
  39. Sehgal, S. A Laboratory Manual of Food Analysis; Wiley: New Delhi, India, 2020. [Google Scholar]
  40. Cunniff, P.A. Official Methods of Analysis of AOAC International, 16th ed.; Association of Official Analysis Chemists: Arlington, MA, USA, 1998. [Google Scholar]
  41. Kang, H.-M.; Saltveit, M.E. Antioxidant capacity of lettuce leaf tissue increases after wounding. J. Agric. Food Chem. 2002, 50, 7536–7541. [Google Scholar] [CrossRef]
  42. Singleton, V.L.; Rossi, J.A. Colorimetry of total phenolics with phosphomolbdic-phosphotungstic acid reagents. Am. J. Enol. Vitic. 1965, 16, 144–158. [Google Scholar]
  43. Kumar, P.; Ramakritinan, C.M.; Kumaraguru, A.K. Solvent extraction and spectrophotometric determination of pigments of some algal species from the shore of puthumadam, southeast coast of India. Int. J. Ocean. Oceanogr. 2010, 4, 29–34. [Google Scholar]
  44. Lichtenthaler, H.K.; Wellburn, A.R. Determinations of Total Carotenoids and Chlorophylls a and b of Leaf Extracts in Different Solvents. Biochem Soc Trans. 1983, 11, 591–592. [Google Scholar] [CrossRef] [Green Version]
  45. Pan-Utai, W.; Iamtham, S. Extraction, purification and antioxidant activity of phycobiliprotein from Arthrospira platensis. Process Biochem. 2019, 82, 189–198. [Google Scholar] [CrossRef]
  46. Basiri, S.; Haidary, N.; Shekarforoush, S.S.; Niakousari, M. Flaxseed mucilage: A natural stabilizer in stirred yogurt. Carbohydr. Polym. 2018, 187, 59–65. [Google Scholar] [CrossRef]
  47. Marth, E.H. Standard Methods for the Examination of Dairy Products, 14th ed.; American Public Health Association: Washington, DC, USA, 1978. [Google Scholar]
  48. Koli, D.K.; Rudra, S.G.; Bhowmik, A.; Pabbi, S. Nutritional, functional, textural and sensory evaluation of Spirulina enriched green pasta: A potential dietary and health supplement. Foods 2022, 11, 979. [Google Scholar] [CrossRef]
  49. Tańska, M.; Konopka, I.; Ruszkowska, M. Sensory, physico-chemical and water sorption properties of corn extrudates enriched with Spirulina. Plant Foods Hum. Nutr. 2017, 72, 250–257. [Google Scholar] [CrossRef] [Green Version]
  50. El-Hameed, M.K.A.; El-Maatti, S.M.A.; El-Saidy, S.M.; Ahmed, S.M. Effect of adding Spirulina platensis in pasta products (spaghetti). Zagazig J. Agric. Res. 2018, 45, 293–300. [Google Scholar] [CrossRef]
  51. Hidayati, J.R.; Yudiati, E.; Pringgenies, D.; Oktaviyanti, D.T.; Kusuma, A.P. Comparative study on antioxidant activities, total phenolic compound and pigment contents of tropical Spirulina platensis, Gracilaria arcuata and Ulva lactuca extracted in different solvents polarity. In E3S Web of Conferences; EDP Sciences: Les Ulis, France, 2020; Volume 147, p. 03012. [Google Scholar] [CrossRef]
  52. Mehta, B.M.; de Oliveira, M.N. 11 Fermented Dairy Products. In Fermentation: Effects on Food Properties; CRC Press: Boca Raton, FL, USA, 2012; p. 259. [Google Scholar]
  53. Ásványi-Molnár, N.; Sipos-Kozma, Z.S.; Tóth, Á.; Ásványi, B.; Varga, L. Development of functional dairy food enriched in spirulina (Arthrospira platensis). Tejgazdasag 2009, 69, 15–22. [Google Scholar]
  54. Gyenis, B.; Szigeti, J.F.; Asvanyi-Molnar, N.; Varga, L. Use of dried microalgal biomasses to stimulate acid production and growth of Lactobacillus plantarum and Enterococcus faecium in milk. Acta Agrar. 2005, 9, 53–59. [Google Scholar]
  55. Varga, L.; Szigeti, J.; Kovács, R.; Földes, T.; Buti, S. Influence of a Spirulina platensis Biomass on the Microflora of Fermented ABT Milks During Storage (R1). J. Dairy Sci. 2002, 85, 1031–1038. [Google Scholar] [CrossRef]
  56. Patel, P.; Jethani, H.; Radha, C.; Vijayendra, S.V.N.; Mudliar, S.N.; Sarada, R.; Chauhan, V.S. Development of a carotenoid enriched probiotic yogurt from fresh biomass of Spirulina and its characterization. J. Food Sci. Technol. 2019, 56, 3721–3731. [Google Scholar] [CrossRef]
  57. Shimamatsu, H. Mass production of Spirulina, an edible microalga. Hydrobiologia 2004, 512, 39–44. [Google Scholar] [CrossRef]
  58. Lupatini, A.L.; Colla, L.M.; Canan, C.; Colla, E. Potential application of microalga Spirulina platensis as a protein source. J. Sci. Food Agric. 2017, 97, 724–732. [Google Scholar] [CrossRef] [PubMed]
  59. Üstün-Aytekin, Ö.; Çoban, I.; Aktaş, B. Nutritional value, sensory properties, and antioxidant activity of a traditional kefir produced with Arthrospira platensis. J. Food Process. Preserv. 2022, 46, e16380. [Google Scholar] [CrossRef]
  60. Khaledabad, M.A.; Ghasempour, Z.; Kia, E.M.; Bari, M.R.; Zarrin, R. Probiotic yoghurt functionalised with microalgae and zedo gum: Chemical, microbiological, rheological and sensory characteristics. Int. J. Dairy Technol. 2019, 73, 67–75. [Google Scholar] [CrossRef]
  61. Ozturkoglu-Budak, S.; Akal, C.; Yetisemiyen, A. Effect of dried nut fortification on functional, physicochemical, textural, and microbiological properties of yogurt. J. Dairy Sci. 2016, 99, 8511–8523. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  62. Fradinho, P.; Niccolai, A.; Soares, R.; Rodolfi, L.; Biondi, N.; Tredici, M.R.; Sousa, I.; Raymundo, A. Effect of Arthrospira platensis (spirulina) incorporation on the rheological and bioactive properties of gluten-free fresh pasta. Algal Res. 2020, 45, 101743. [Google Scholar] [CrossRef]
  63. Ismaiel, M.M.S.; El-Ayouty, Y.M.; Piercey-Normore, M. Role of pH on antioxidants production by Spirulina (Arthrospira) platensis. Braz. J. Microbiol. 2016, 47, 298–304. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  64. Goiris, K.; Muylaert, K.; Fraeye, I.; Foubert, I.; De Brabanter, J.; De Cooman, L. Antioxidant potential of microalgae in relation to their phenolic and carotenoid content. J. Appl. Phycol. 2012, 24, 1477–1486. [Google Scholar] [CrossRef]
  65. Anbarasan, V.; Kishor Kumar, V.; Satheesh Kumar, P.; Venkatachalam, T. In vitro evaluation of antioxidant activity of blue green algae S. platensis. Int. J. Pharm. Sci. Res. 2011, 2, 2616–2618. [Google Scholar]
  66. Marzorati, S.; Schievano, A.; Idà, A.; Verotta, L. Carotenoids, chlorophylls and phycocyanin from Spirulina: Supercritical CO2 and water extraction methods for added value products cascade. Green Chem. 2019, 22, 187–196. [Google Scholar] [CrossRef]
  67. Shimamatsu, H. A pond for edible Spirulina production and its hydraulic studies. In Twelfth International Seaweed Symposium; Springer: Dordrecht, The Netherlands, 1987; pp. 83–89. [Google Scholar] [CrossRef]
  68. Szmejda, K.; Duliński, R.; Byczyński, Ł.; Karbowski, A.; Florczak, T.; Żyła, K. Analysis of the selected antioxidant compounds in ice cream supplemented with Spirulina (Arthrospira platensis) extract. Biotechnol. Food Sci. 2018, 82, 41–48. [Google Scholar]
  69. Khandual, S.; Sanchez, E.O.L.; Andrews, H.E.; de la Rosa, J.D.P. Phycocyanin content and nutritional profile of Arthrospira platensis from Mexico: Efficient extraction process and stability evaluation of phycocyanin. BMC Chem. 2021, 15, 24. [Google Scholar] [CrossRef]
  70. Rito-Palomares, M.; Nuñez, L.; Amador, D. Practical application of aqueous two-phase systems for the development of a prototype process for c-phycocyanin recovery from Spirulina maxima. J. Chem. Technol. Biotechnol. 2001, 76, 1273–1280. [Google Scholar] [CrossRef]
  71. Niccolai, A.; Shannon, E.; Abu-Ghannam, N.; Biondi, N.; Rodolfi, L.; Tredici, M.R. Lactic acid fermentation of Arthrospira platensis (spirulina) biomass for probiotic-based products. J. Appl. Phycol. 2019, 31, 1077–1083. [Google Scholar] [CrossRef] [Green Version]
  72. Liu, J.-G.; Hou, C.-W.; Lee, S.-Y.; Chuang, Y.; Lin, C.-C. Antioxidant effects and UVB protective activity of Spirulina (Arthrospira platensis) products fermented with lactic acid bacteria. Process Biochem. 2011, 46, 1405–1410. [Google Scholar] [CrossRef]
  73. Güldas¸, M.; Irkin, R. Influence of Spirulina platensis powder on the microflora of yoghurt and acidophilus milk. Mljekarstvo 2010, 60, 237–243. [Google Scholar]
  74. Sudheendra, C.V.K. Development of Synbiotic Lassi Added with Carrot Juice. Ph.D. Thesis, Agricultural University, Anand, India, 2010. [Google Scholar]
  75. Akalin, A.; Unal, G.; Dalay, M. Influence of Spirulina platensis biomass on microbiological viability in traditional and probiotic yogurts during refrigerated storage. Ital. J. Food Sci. 2009, 21, 357–364. [Google Scholar]
  76. De Caire, G.Z.; Parada, J.L.; Zaccaro, M.C.; De Cano, M.M.S. Effect of Spirulina platensis biomass on the growth of lactic acid bacteria in milk. World J. Microbiol. Biotechnol. 2000, 16, 563–565. [Google Scholar] [CrossRef]
  77. Gupta, S.; Gupta, C.; Garg, A.P.; Prakash, D. Prebiotic efficiency of blue green algae on probiotics microorganisms. J. Microbiol. Exp. 2017, 4, 00120. [Google Scholar] [CrossRef] [Green Version]
Dairy 04 00019 g001 550
Figure 1. Changes in titratable acidity and pH drop in T0 (A), T1 (B), T2 (C), and T3 (D) during fermentation period (4 h).
Figure 1. Changes in titratable acidity and pH drop in T0 (A), T1 (B), T2 (C), and T3 (D) during fermentation period (4 h).
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Figure 2. Sensory evaluation of control and different spirulina-treated samples in hedonic scale.
Figure 2. Sensory evaluation of control and different spirulina-treated samples in hedonic scale.
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Figure 3. The pH (A) and acidity (B) value, L. bulgaricus (C) and S. thermophilus (D) counts (log cfumL−1) expressed in (mean ± SD) of samples during 21 days of storage at 4 °C.
Figure 3. The pH (A) and acidity (B) value, L. bulgaricus (C) and S. thermophilus (D) counts (log cfumL−1) expressed in (mean ± SD) of samples during 21 days of storage at 4 °C.
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Table
Table 1. Nutritional composition, antioxidant activity, and total phenolic content of the procured Spirulina powder.
Table 1. Nutritional composition, antioxidant activity, and total phenolic content of the procured Spirulina powder.
ComponentsValue (%)
Moisture5.12 ± 0.03
Protein59.27 ± 0.02
Total solids94.88 ± 0.03
Ash contents7.02 ± 0.03
% DPPH inhibition89.00 ± 1.02
Total Phenolic Content (TPC)11.26 ± 0.04 mg GAE/100 g
Results are expressed as the mean ± standard deviation (n = 3).
Table
Table 2. Physicochemical Characteristics of buttermilk samples.
Table 2. Physicochemical Characteristics of buttermilk samples.
ParametersT0T1p Value
Total Solids (g/100 g on dry weight basis)9.73 ± 0.019.87 ± 0.01 b0.004
Protein (g/100 g on dry weight basis)1.71 ± 0.021.83 ± 0.01 b0.003
Fat (g/100 g on dry weight basis)0.70 ± 0.010.733 ± 0.020.065
Carbohydrate (g/100 g on dry weight basis)6.56 ± 0.016.59 ± 0.02 b0.001
Energy (Kcal/100 mL)39.45 ± 0.1840.29 ± 0.11 b0.007
Ash (g/100 g on dry weight basis)0.70 ± 0.020.76 ± 0.03 a0.042
Calcium (mg/100 g on dry weight basis)74.59 ± 0.9782.06 ± 1.02 b0.001
Viscosity (cP)20.2 ± 0.0121.9 ± 0.01 b0.008
Results are expressed as the mean ± standard deviation (n = 3). The values followed by superscript letters (a and b) are significantly different between treatment means at a p < 0.05; b p < 0.01. Control (T0) vs. 0.25% Spirulina-fortified buttermilk (T1).
Table
Table 3. Antioxidant parameters of control and Spirulina incorporated buttermilk.
Table 3. Antioxidant parameters of control and Spirulina incorporated buttermilk.
Parameters T0T1p Value
DPPH–scavenging activity (%)41.99 ± 0.8148.19 ± 0.26 a<0.001
Chlorophyll (mg/g on dry weight basis)0.30 ± 0.0230.91 ± 0.04 a<0.001
Carotenoid (mg/g on dry weight basis)08.24 ± 0.02 a<0.001
C-Phycocyanin (mg/g on dry weight basis)00.30 ± 0.01 a<0.001
Total Phenolic Content (mg/g GAE)2.44 ± 0.014.21 ± 0.03 a<0.001
Results are expressed as the mean ± standard deviation (n = 3). Values with superscript letters are significantly different at p < 0.001.
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Rose, H.; Bakshi, S.; Kanetkar, P.; Lukose, S.J.; Felix, J.; Yadav, S.P.; Gupta, P.K.; Paswan, V.K. Development and Characterization of Cultured Buttermilk Fortified with Spirulina plantensis and Its Physico-Chemical and Functional Characteristics. Dairy 2023, 4, 271-284. https://doi.org/10.3390/dairy4020019

AMA Style

Rose H, Bakshi S, Kanetkar P, Lukose SJ, Felix J, Yadav SP, Gupta PK, Paswan VK. Development and Characterization of Cultured Buttermilk Fortified with Spirulina plantensis and Its Physico-Chemical and Functional Characteristics. Dairy. 2023; 4(2):271-284. https://doi.org/10.3390/dairy4020019

Chicago/Turabian Style

Rose, Hency, Shiva Bakshi, Prajasattak Kanetkar, Smitha J. Lukose, Jude Felix, Satya Prakash Yadav, Pankaj Kumar Gupta, and Vinod Kumar Paswan. 2023. "Development and Characterization of Cultured Buttermilk Fortified with Spirulina plantensis and Its Physico-Chemical and Functional Characteristics" Dairy 4, no. 2: 271-284. https://doi.org/10.3390/dairy4020019

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