Production and quality characterizations of probiotic-germinated rice beverage

Production and quality characterizations of probiotic-germinated rice beverage | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Production and quality characterizations of probiotic-germinated rice beverage Prinya Wongsa, Natee Charleeniwat This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8117321/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Probiotic cereal-based beverages have gained increasing attention as novel functional and fast-growing products. This study developed a probiotic beverage from germinated pigmented rice-based powder enriched with probiotics, specifically Bifidobacterium animalis subsp. lactis (BB-12®) and Lactobacillus paracasei (L. casei 431®). Indigenous polished and pigmented rice varieties from Thailand were subjected to germination to enhance their bioactive compound content and biological activities. Among the tested varieties, the pigmented rice cultivar Riceberry exhibited the highest concentrations of γ-aminobutyric acid, total phenolic content, antioxidant activity, and acetylcholinesterase and butyrylcholinesterase inhibitory activities. Riceberry flour and other ingredients were used to produce a pigmented cereal-based powder, which was subsequently fortified with freeze-dried probiotic powder at mixing ratios of 3% and 5%. The probiotic pigmented cereal-based powder and its corresponding beverage achieved viable probiotic counts above 6 log cfu/mL, meeting the minimum threshold for probiotic efficacy. The beverage also demonstrated favourable sensory attributes, particularly in colour, aroma, and flavour. Furthermore, probiotic-germinated Riceberry powder packaged in vacuum-sealed aluminium pouches remained shelf-stable under room-temperature storage, maintaining high probiotic viability and good quality for up to 6 months. Pigmented rice germination probiotics bioactive compounds functional drinks Figures Figure 1 Figure 2 1 Introduction In recent years, the rapid development of functional foods and beverages has attracted increasing attention from both the food industry and the public for their potential to promote human health [ 1 ]. Intentionally modified functional foods, including the probiotication of foods—enriched with probiotics such as lactic acid bacteria (LAB) in adequate amounts—aim to provide scientifically proven benefits to support gut health [ 2 ]. Probiotics are living microorganisms that, when administered in sufficient quantities, confer a health benefit on the host. According to World Health Organization/Food and Agriculture Organization recommendations, any food claiming a probiotic effect must contain at least 6 log cfu/mL to 7 log cfu/mL (10 6 to 10 7 cfu/mL) of viable probiotic bacteria [ 3 ]. LAB, including members of the Lactobacillus and Bifidobacterium genera, exhibit probiotic properties. These genera are part of the indigenous microbiota of humans and animals and are generally recognised as safe [ 4 ]. Consequently, there has been an expansion in the application of probiotics not only in nutraceuticals but also as functional ingredients in foods [ 5 ]. Several studies have reported that cereal-based ingredients—such as maize, millet, barley, oat, rye, wheat, and rice—fermented with probiotics can enhance consumer health by combining the benefits of probiotics with health-promoting bioactive components [ 6 , 7 ]. Rice ( Oryza sativa L.) is a staple food for most of the global population because of its nutritional value, quality characteristics, and diverse varieties that meet consumer demand. In 2023, the global harvested rice area was 168,356,566 hectares, while total rice production reached 799,999,504.87 tonnes [ 8 ]. White or polished rice is widely consumed, whereas pigmented varieties such as black, red, and purple rice are preferred by certain consumer groups. Pigmented rice accumulates bioactive metabolites, including cyanidin-3-glucoside, cyanidin-3-galactoside, malvidin-3-galactoside, and peonidin-3-glucoside. In addition, bioactive compounds in pigmented rice offer potential health benefits, including antioxidant, anti-inflammatory, anti-cancer, and anti-diabetic activities [ 9 ]. In Thailand, a purple rice cultivar known as Riceberry—a novel variety rich in antioxidant properties and other bioactive compounds—is available to consumers [ 10 ]. Furthermore, bioactive compounds such as gamma-aminobutyric acid (GABA) in cereal grains can be enhanced through germination, a process influenced by factors such as temperature, time, pH, processing method, and drying conditions that affect biochemical reactions within plant cells. Germination increases the bioavailability of nutrients such as carbohydrates, proteins, GABA, and antioxidants, while reducing anti-nutritional factors such as phytates [ 11 ]. Moreover, germinated rice exhibits improved sensory qualities, including sweetness, texture, and ease of cooking. Therefore, germination plays a crucial role in enhancing the nutritional value and overall quality of brown rice [ 12 ]. Consequently, germinated rice grains can be utilised as a pre-prepared ingredient for developing value-added products such as rice flour, rice milk, rice starch, rice puddings, and rice cakes [ 13 ]. Powdered food ingredients, such as spray-dried and freeze-dried powders, are widely produced by the food industry because they are easier to process, handle, transport, and store. In addition, instant food powders offer consumers convenient, minimal-preparation options and can be formulated with functional ingredients [ 14 ]. Various types of instant foods—such as instant germinated purple rice tea [ 15 ] and black rice-based instant beverage mixes [ 16 ]—have been developed using healthy or functional ingredients. Moreover, the enrichment of instant food powders with LAB strains has been explored. Lipan et al [ 17 ] developed spray-dried almond milk powder containing Lactobacillus plantarum (ATCC 8014) at concentrations above 10 7 viable cells, maintaining potential probiotic functionality for up to 8 months when stored at 4°C and 6 months at 22°C. However, spray drying at high temperatures remains disadvantageous for drying bioactive or heat-sensitive compounds such as enzymes, cells, and vitamins [ 18 ]. Additionally, designing non-dairy instant powders, such as cereal-based powders, can be challenging because of their macromolecular composition, structural complexity, and the high degree of formulation flexibility required. Rittisak et al. [ 10 ] developed an instant beverage using broken Riceberry rice through the application of a double drum dryer operated at 125°C and a drum speed of 1.0 rpm. The resulting dried powder exhibited a light purple colour, low water activity (0.354), and minimal microbial contamination, with total plate counts and yeast and mould levels below 10 cfu/g. The formulation and process for enriching plant-based products with phenolic compounds, dietary fibres, and/or probiotics should be carefully tailored to achieve the desired quality characteristics and improve viable probiotic counts [ 19 ] Because instant powdered beverages typically require rehydration before consumption, their reconstitution ability—along with the retention of nutrients, functional compounds, and associated health benefits—is a crucial techno-functional property for both production and storage [ 20 ]. Despite growing interest in functional beverages, the development of probiotic cereal drinks and the effects of probiotic enrichment on viable cell counts have not been extensively studied. Furthermore, the existing literature offers limited information in this area, particularly regarding the development of beverages using germinated pigmented rice as an ingredient combined with novel probiotication techniques. Therefore, the objectives of this study were to develop a novel probiotic beverage from germinated pigmented rice powder and to evaluate the viability of probiotics such as Bifidobacterium animalis subsp. lactis (BB-12®) and Lactobacillus paracasei (L. casei 431®) following beverage preparation and storage. 2 Materials and methods 2.1 Plant materials Rice ( Oryza sativa L.) cultivars Hom Mali 105, RD6, Khiao Ngu, and Riceberry were obtained from a local market in Chiang Rai Province, Thailand. 2.2 Lactic acid cultures Lactic acid-forming bacteria, including Bifidobacterium animalis subsp. lactis (BB-12®) and Lactobacillus paracasei (L. casei 431®), were obtained as Direct Vat Set cultures (10-pouch, 50 U size) from Chr. Hansen, Hoersholm, Denmark. 2.3 Chemicals All chemicals used in this study were of analytical reagent grade, mainly manufactured by Sigma-Aldrich (Chemie GmbH, Germany) and supplied by local chemical distributors in Thailand. Purified water was produced using a Milli-Q purification system (Merck Millipore, Bedford, MA, USA). 2.4 Germination Twenty grams of paddy rice were washed with tap water and soaked in distilled water for 12 hours. After soaking, the samples were rinsed three times with distilled water and drained. The soaked rice grains were then placed in germination boxes lined with clean, sterile cloth and kept in a dark place at 30°C for 24, 48, 72, 96, and 120 hours. At each designated germination time, the samples were washed with distilled water and freeze-dried. The dried samples were ground into a 60-mesh powder and stored in sealed plastic bags at 4°C for further analysis of bioactive compounds and bioactivities. The active compounds and bioactivities—including GABA, total phenolic compounds, antioxidant activity, and cholinesterase enzyme inhibition—were evaluated in the germinated rice and used as selection criteria for developing the probiotic rice beverage. 2.5 Production of germinated rice-based flake The production of the cereal flake was carried out based on a preliminary formulation (data not shown). The optimised recipe consisted of germinated Riceberry flour (29.4%) combined with dry ingredients, including inulin (20.0%), whey protein (15.0%), maltodextrin (10.0%), corn-based milk powder alternative (10.0%), full-cream milk powder (5.0%), malt powder (5.0%), and sucralose (0.1%). The dry mixture was then blended with reverse osmosis water at a mixing ratio of 1:2 (w/w). Xanthan gum (0.5%) was subsequently added. The mixture was homogenised using a KitchenAid stand mixer (Artisan®, USA) and then dried using a twin drum dryer (Euro Best Technology Co., Ltd., Bangkok, Thailand) at 150°C with a roller rotation speed of 1 rpm. The resulting germinated Riceberry flake was collected and cooled to room temperature. The cereal flakes were then ground in a cooling-assisted grinder and stored at room temperature for subsequent development into probiotic-germinated Riceberry powder. 2.6 Production of freeze-dried probiotic powder Probiotic cultures, including BB-12® and L. casei 431®, were inoculated to achieve an initial viable bacterial level of at least 6 log cfu/g. The liquid milk was pasteurised at 73°C for 15 seconds and then cooled to 43°C–45°C. The commercial freeze-dried starter was subsequently added and mixed thoroughly for 10–15 minutes. Fermentation was carried out at 45°C for 24 hours. After fermentation, the yoghurt was freeze-dried at a shelf temperature of 20°C using a laboratory-scale freeze dryer (Martin Christ Gefriertrocknungsanlagen GmbH, Germany). The resulting lyophilised flakes were ground in a cooling-assisted grinder and stored at room temperature before being mixed with germinated Riceberry powder. 2.7 Production of probiotic-germinated rice powder Germinated Riceberry powder (see Section 2.5 ) was prepared and fortified with freeze-dried BB-12® and L. casei 431® powders (see Section 2.6 ) at mixing ratios of 3% (w/w) and 5% (w/w), respectively. A 30-g portion of the probiotic cereal powder was packed in a laminated aluminium pouch with a low-density polyethylene (LDPE/AL/LDPE) lining (10- × 15-cm size, 110-µm thickness). The shelf-life study of the samples was conducted at 35°C for a storage period of 6 months. 2.8 Determination of GABA content in germinated rice The analysis of GABA content was carried out following the method of Kamjijam et al [ 21 ] with slight modifications. A 0.5-g sample was placed in a test tube and mixed with 0.9 mL of distilled water and 0.9 mL of ethanol containing 3% sulfosalicylic acid (v/v). The solution was stirred thoroughly and centrifuged at 5,000 rpm for 10 minutes. An aliquot of the supernatant (50 µL) was then mixed with 50 µL of NaHCO 3 solution and 200 µL of DABSYL-Cl acetonitrile solution. The mixture was incubated at 70°C for 10 minutes, followed by the addition of 250 µL of ethanol and 250 µL of KH 2 PO 4 buffer solution (pH 6.8). The mixture was then centrifuged at 4,500 rpm for 10 minutes and filtered through a 0.45-µm nylon membrane filter. The resulting solution was analysed using an Acquity™ UPLC system (Waters Acquity™; Waters Corp., Milford, MA, USA) equipped with a BEH C18 column (100 mm × 2.1 mm, 1.7 µm; Waters Corp.) and a UV detector set at 465 nm. 2.9 Determination of total phenolic content (TPC) The TPC was determined using the Folin–Ciocalteu assay [ 22 ]. An aliquot of the germinated rice extract (20 µL) was mixed with 100 µL of 10% Folin–Ciocalteu reagent and 80 µL of 7.5% sodium carbonate solution. The mixture was incubated in the dark at room temperature for 30 minutes. The absorbance was then measured at 760 nm using a UV–visible spectrophotometer (Biochrom Libra S22, UK). TPC was calculated using a gallic acid standard curve, and the results were expressed as milligrams of gallic acid equivalents per 100 g dry matter (mg GAE/100 g DM). 2.10 Determination of antioxidant activity The antioxidant activity was determined using the DPPH radical scavenging assay, following the method described by Netrprachit et al. [ 22 ] with slight modifications. An extract (50 µL) was mixed with 100 µL of DPPH solution in methanol (5.9 mg/100 mL, w/v) in a 96-well plate. The mixture was then incubated in the dark for 30 minutes. The absorbance was measured at 517 nm using a 96-well plate UV–Vis spectrophotometer (Thermo Fisher Scientific Inc., MA, USA). The control consisted of distilled water (250 µL) in place of the extract. The percentage of antioxidant activity was then calculated by comparing the absorbance of the sample to that of the control. 2.11 Determination of inhibitory potential against cholinesterase enzymes The determination of acetylcholinesterase (AChE) inhibition in the germinated rice extract was carried out following the method of Temviriyanukul et al. [ 23 ] with slight modifications. An extract (50 µL) was mixed with an enzyme solution consisting of Electrophorus electricus AChE (20 ng; 1000 units/mg, 100 µL) in 50 mM potassium phosphate buffer (KPB, pH 7.0) in a 96-well plate. Subsequently, 10 µL of 16 mM 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB) and 40 µL of 0.8 mM acetylthiocholine in 50 mM KPB (pH 7.0) were added to the mixture. The absorbance was measured at 412 nm using a 96-well plate UV–Vis spectrophotometer (Thermo Fisher Scientific Inc.). The percentage of inhibition was then calculated. The determination of butyrylcholinesterase (BChE) inhibition was performed using the same procedure. A solution containing 100 ng of equine serum BChE (≥ 10 units/mg protein, 100 µL) in 50 mM KPB (pH 7.0) with 1 mM MgCl 2 and 0.1 mM butyrylthiocholine (BTCh) in 50 mM KPB (pH 7.0) served as the reactive enzyme–substrate mixture. The absorbance was recorded at 412 nm using a 96-well plate UV–Vis spectrophotometer (Thermo Fisher Scientific Inc.), and the percentage of inhibition was calculated accordingly. 2.12 Enumeration of probiotic cultures Enumeration of LAB was performed using the 3M Petrifilm® Lactic Acid Bacteria Count Plate method [ 24 ] (3M Inc., MN, USA; https://www.3m.com ). The number of colonies was expressed as log 10 cfu/g. 2.13 Enumeration of viable probiotic in geminated Riceberry beverage A 15-g sample of probiotic-germinated Riceberry powder was dissolved in 150 mL of warm water at 60°C, 80°C, and 100°C, with stirring for 3 minutes to ensure complete dissolution. The prepared beverages were then transferred into 50-mL sealed plastic tubes and stored at − 20°C for subsequent microbial colony analysis. 2.14 Enumeration of viable probiotic in geminated rice beverage during storage Samples were collected every 30 days during storage to determine the viable probiotic count, [ 25 ] as described in Section 2.13 . 2.15 Determination physicochemical property of geminated rice beverage during storage Water activity (Aw) was measured using an Aw meter (AQUALAB Model Series 3TE; AquaLab, Pullman, WA, USA). Moisture content was determined according to the AOAC method 925.10, [ 25 ] as established by the Association of Official Analytical Chemists. In addition, the water solubility index (WSI) and water absorption index (WAI) were determined following the method of Kim et al. [ 26 ]. 2.16 Sensory evaluation of probiotic-germinated rice beverage Sensory evaluation was conducted with 30 trained panellists (aged 30–45 years), recruited from students and staff of the School of Agro-Industry, Mae Fah Luang University. A 15-g sample of probiotic-germinated rice powder was dissolved in 150 mL of warm water at 80°C and stirred for 3 minutes to ensure complete dissolution. The test samples were served in 15-mL glass cups at room temperature. Panellists evaluated and scored the sensory attributes—appearance, colour, odour, taste, viscosity, and overall liking—using a 9-point hedonic scale [ 27 ] (1 = dislike very much, 9 = like very much). 2.17 Statistical analysis All data were statistically analysed using analysis of variance, and mean comparisons were performed using Duncan’s multiple range test to determine significant differences between samples at a 95% confidence level ( p < 0.05). Statistical analyses were carried out using IBM SPSS® Statistics version 21.0.0 (IBM Corp., Armonk, NY, USA). 3 Results and Discussion 3.1 GABA content in germinated rice The germination of brown rice can improve its flavour, enhance its nutritional value, and increase its potential health benefits. Germinated brown rice is richer in nutrients and bioactive compounds such as essential amino acids, phenolic acids, γ-oryzanol, and GABA [ 28 ]. Therefore, germinated brown rice is increasingly recognised as a promising health food. In this study, germination was performed on four rice varieties: Hom Mali 105, Riceberry, glutinous RD6, and Khiao Ngu. As shown in Fig. 1 , the GABA content in the germinated rice samples changed significantly ( p < 0.05) during the 120-hour germination period. Non-waxy rice varieties, including Hom Mali 105 and Riceberry, exhibited higher GABA levels than did waxy varieties such as RD6 and Khiao Ngu. The GABA content ranged from 4.78 to 16.41 g/100 g DM for Hom Mali 105 and from 7.93 to 26.01 g/100 g DM for Riceberry. The maximum GABA accumulation occurred between 48 and 72 hours of germination for both Hom Mali 105 and Riceberry, after which levels plateaued or declined. Chaijan and Panpipat [ 29 ] reported that the optimal germination time for indigenous Southern Thai rice varieties, including Khemtong and Khai Mod Rin, was 96 hours, resulting in the highest levels of GABA, thiamine, free amino acids, total sugars, and α-amylase activity. The increase in GABA content is likely due to the activity of the enzyme glutamate decarboxylase, which catalyses the conversion of glutamate to GABA [ 30 ]. Similarly, Cáceres et al. [ 31 ] reported that GABA synthesis occurs in a time-dependent manner during germination. 3.2 TPC of germinated rice Longer germination periods resulted in an increase in the TPC of the rice samples (Table 1 ). A significant difference ( p < 0.05) in TPC was observed at all 24-hour intervals during germination. Germinated Riceberry showed the highest TPC, increasing from 163.43 mg GAE/100 g DW at 0 h to 299.53 mg GAE/100 g DW at 120 h. Similarly, Singh et al. (2017) [ 32 ] reported that germination of non-waxy Thai rice cultivar Phitsanulok 2 at 33°C for 300 minutes increased both GABA and TPC levels—from 6.71 mg/100 g and 13.68 mg GAE/100 g DM to 18.74 mg/100 g and 33.95 mg GAE/100 g DM, respectively. Structural changes in the grain during germination and mashing are related to various enzymatic degradations that facilitate the release and extraction of free polyphenols [ 33 ] Enzyme activities may also vary considerably depending on temperature and pH conditions. Table 1 Total phenolic content of germinated rice Germinated rice extract Total phonic content (mg GAE/100 g DW) 0 h 24 h 48 h 72 h 96 h 120 h Hom Mali 105 65.31 ± 5.31 b 74.03 ± 3.35 b 88.16 ± 6.43 b 110.13 ± 5.20 b 98.15 ± 4.59 b 109.83 ± 5.77 b Riceberry 163.43 ± 3.53 a 170.11 ± 4.67 a 184.74 ± 8.67 a 200.84 ± 3.42 a 257.91 ± 3.45 a 299.53 ± 5.77 a RD6 43.21 ± 1.33 c 47.71 ± 2.71 c 48.46 ± 0.87 c 53.08 ± 1.34 c 55.29 ± 3.52 c 58.34 ± 2.89 c Khiao Ngu 16.39 ± 0.28 d 17.34 ± 0.35 d 14.07 ± 0.17 d 20.41 ± 0.12 d 17.09 ± 0.24 d 19.83 ± 0.77 d Data are presented as mean ± SD of three measurements (n = 3). Means in columns with superscript letters indicate significant differences ( p < 0.05). 3.3 Antioxidant activity of germinated rice Table 2 presents the antioxidant activity of 50 µg/mL extracts from both ungerminated and germinated rice samples. The initial antioxidant activity of the ungerminated rice samples showed significant differences ( p < 0.05). Among them, the Riceberry extract exhibited the highest percentage of DPPH inhibition (35.07% ± 0.89%). The antioxidant activity of non-pigmented rice varieties was comparatively lower than that of pigmented rice. Black rice varieties are rich in anthocyanins, primarily cyanidin-3-glucoside and peonidin-3-glucoside, which range from 19.4 to 140.8 mg/100 g DM and 11.1 to 12.8 mg/100 g DM, respectively [ 9 ]. The biological functionality of pigmented rice is largely attributed to its polyphenolic compounds. Furthermore, the antioxidant activity of all germinated extracts increased significantly ( p < 0.05) with longer germination periods, indicating that germination positively affects antioxidant properties. Several studies have reported similar findings, showing that the TPC of germinated black waxy rice is higher than that of non-germinated rice, while germinated red jasmine rice exhibits slightly lower values. Phattayakorn et al. [ 34 ] also reported that germinated rice demonstrated higher antioxidant activity than non-germinated rice, as determined by FRAP and DPPH assays. Table 2 Antioxidant activity (DPPH scavenging) of germinated rice Germinated rice extract Antioxidant potential (%) of 50 µg/mL germinated rice extract 0 h 24 h 48 h 72 h 96 h 120 h Hom Mali 105 9.34 ± 0.89 b 9.07 ± 0.23 b 10.72 ± 0.12 b 11.54 ± 0.51 b 12.17 ± 0.79 b 14.08 ± 0.4 b Riceberry 35.07 ± 0.89 a 37.29 ± 0.91 a 47.57 ± 1.25 a 56.31 ± 1.93 a 65.46 ± 2.51 a 71.23 ± 0.11 a RD6 5.54 ± 0.08 c 6.74 ± 0.11 c 7.14 ± 0.59 c 9.03 ± 0.28 b,c 9.52 ± 0.85 b 10.15 ± 0.92 b Khiao Ngu 6.17 ± 0.19 c 7.09 ± 0.08 c 7.81 ± 0.19 c 8.07 ± 0.23 c 10.17 ± 0.08 b 10.72 ± 0.39 b Data are presented as mean ± SD of three measurements (n = 3). Means in columns with superscript letters indicate significant differences ( p < 0.05). 3.4 AChE and BChE inhibitory potential of germinated rice The extracts of germinated rice demonstrated inhibitory potential against cholinesterase enzyme activities (Tables 3 and 4 ). Germinated pigmented rice cultivar Riceberry showed higher AChE and BChE inhibitory activities than germinated non-pigmented rice at all germination periods. In comparison, the extract of parboiled germinated brown rice exhibited 6% AChE inhibition and 39% BChE inhibition, with significantly higher inhibitory activity than other rice types, including germinated brown rice, brown rice, and white rice [ 35 ]. The enhanced inhibitory effects may be attributed to bioactive peptides generated during rice germination and mashing, which can contribute to angiotensin-converting enzyme, AChE, and BChE inhibitory capacities [ 29 ]. Table 3 AChE inhibitory potential of germinated rice Germinated rice extract AChE inhibitory potential of 50 µg/mL germinated rice extract 0 h 24 h 48 h 72 h 96 h 120 h Hom Mali 105 6.09 ± 0.27 b 6.99 ± 0.17 b 8.19 ± 0.72 b 14.71 ± 3.7 b 17.86 ± 1.33 b 18.79 ± 0.45 b Riceberry 26.09 ± 0.72 a 24.09 ± 0.72 a 26.09 ± 0.72 a 24.71 ± 3.77 a 26.86 ± 1.33 a 28.79 ± 0.45 a RD6 4.00 ± 0.13 c 3.91 ± 1.22 c 5.49 ± 1.45 c 3.17 ± 0.56 c 3.99 ± 0.45 c 4.41 ± 0.34 c Khiao Ngu 3.88 ± 0.33 c 4.31 ± 0.89 c 3.99 ± 0.65 c 4.45 ± 0.62 c 4.78 ± 1.04 c 4.51 ± 0.98 c Data are presented as mean ± SD of three measurements (n = 3). Means in columns with superscript letters indicate significant differences ( p < 0.05). Table 4 BChE inhibitory potential of germinated rice Germinated rice extract BChE inhibitory potential of 50 µg/mL germinated rice sample extract 0 h 24 h 48 h 72 h 96 h 120 h Hom Mali 105 35.19 ± 1.50 b 33.90 ± 1.78 b 37.81 ± 1.73 b 40.34 ± 0.89 b 41.09 ± 1.92 b 41.98 ± 2.45 b Riceberry 41.91 ± 0.50 a 43.91 ± 1.20 a 51.08 ± 0.87 a 55.75 ± 0.23 a 59.74 ± 0.51 a 59.80 ± 0.45 a RD6 26.43 ± 1.03 bc 25.82 ± 0.88 bc 30.21 ± 1.57 bc 28.76 ± 0.23 bc 30.32 ± 1.23 bc 31.53 ± 1.99 bc Khiao Ngu 18.12 ± 0.81 c 19.23 ± 1.48 c 20.22 ± 0.87 c 21.61 ± 1.12 c 22.32 ± 0.78 c 21.53 ± 0.60 c Data are presented as mean ± SD of three measurements (n = 3). Means in columns with superscript letters indicate significant differences ( p < 0.05). 3.5 Colony counts of probiotic starter in pre-mixed powder The colony counts of probiotic starters L. casei 431® and BB-12® in the pre-mixed powder were 8.95 and 8.92 log 10 cfu/g, respectively (Table 5 ). Freeze-drying effectively preserves probiotic viability, maintaining the strains’ original chemical composition and physical properties, while enabling long-term storage and good fermentation performance [36 − 7]. Table 5 Colony counts of probiotic starter in pre-mixed powder Starter LAB (log 10 /g) L. paracasei subs. paracasei, L. casei 431® 8.95 B. animalis subsp. lactis, BB-12® 8.92 LAB, lactic acid bacteria 3.6 Production of probiotic-germinated Riceberry powder Germination of Riceberry for 120 hours resulted in the highest levels of GABA, TPC, antioxidant activity, and AChE and BChE inhibitory potentials ( p < 0.05). Therefore, germinated Riceberry was selected as the raw material for preparing germinated rice-based flakes (Section 2.5 ) and subsequently used to produce probiotic-germinated Riceberry powder (Section 2.7 ). 3.7 Viable probiotic counts in geminated Riceberry beverage Beverages prepared from germinated Riceberry-based powder mixed with L. casei 431® and BB-12® at mixing ratios of 3% and 5% (w/w) were reconstituted with warm water at 60°C, 80°C, and 100°C and analysed for LAB counts (Table 6 ). The results showed that the viability of both probiotic strains decreased slightly at both mixing ratios. Increasing the reconstitution temperature led to a reduction in viable counts of L. casei 431® and BB-12® in the probiotic beverages. The final counts decreased by 0.17–0.21 log 10 /mL for L. casei 431® and 0.14–0.24 log cfu/mL for BB-12®. The percentage of reduction ranged from 31.38% to 58.68% for L. casei 431® and BB-12®, respectively, when beverages were prepared with warm water at 60°C–80°C. Notably, probiotic beverages prepared at 60°C and 80°C still retained viable counts above 6 log 10 /mL for both strains. This indicates that the viable counts of L. casei 431® and BB-12® in the novel germinated Riceberry beverages prepared at 60°C and 80°C met the minimum threshold of 6–7 log cfu/mL required for foods claiming probiotic functionality [ 3 ]. Table 6 Viable probiotic counts in geminated Riceberry beverage Probiotic beverage powder Mixing ratio (% w/w) Warm water temperature (°C) LAB (log 10 /mL) Germinated Riceberry-based powder mixed with L. casei 431® 3 60 6.22 80 6.05 100 5.84 5 60 6.57 80 6.40 100 6.23 Germinated Riceberry-based powder mixed with BB-12® 3 60 6.25 80 6.08 100 5.92 5 60 6.67 80 6.43 100 6.29 LAB, lactic acid bacteria 3.8 Physicochemical property of probiotic geminated Riceberry powder during storage Table 7 presents the physicochemical properties of germinated Riceberry powder during 6 months of storage. Probiotic-germinated Riceberry powder stored under vacuum packaging showed moisture content and water activity values ranging from 5.31% to 5.42% and 0.410 to 0.435, respectively. For reference, intermediate-moisture foods typically have water activity values between 0.60 and 0.85 and moisture contents ranging from 15% to 40%, which provide stability against microbial contamination and lipid oxidation [ 37 ]. In this study, water activity in the vacuum-packaged samples did not differ significantly ( p ≥ 0.05) throughout storage. Although water activity increased slightly with longer storage duration, it remained below 0.6, indicating that the powder was stable against undesirable chemical reactions and microbial growth. Furthermore, storage duration influenced the WSI and WAI of the probiotic-germinated Riceberry powder. The powder exhibited high solubility, with WSI values ranging from 53.09% to 49.65% for L. casei 431® and from 52.89% to 50.12% for BB-12®, both of which were largely retained during storage. The WAI of the probiotic powder gradually decreased during storage and was considerably low (29%) in the final month of the storage period. Table 7 Physicochemical properties of geminated Riceberry powder during storage Probiotic powder Storage period (months) Moisture content (%) Aw (-) WSI (%) WAI Germinated Riceberry-based powder mixed with L. casei 431® 0 5.31 ± 0.02 ns 0.410 ± 0.00 c 53.09 ± 0.12 a 11.55 ± 0.34 a 3 5.34 ± 0.01 ns 0.416 ± 0.00 b 51.45 ± 0.02 b 9.25 ± 0.52 b 6 5.38 ± 0.05 ns 0.430 ± 0.00 a 49.65 ± 0.16 c 9.01 ± 0.23 c Germinated Riceberry-based powder mixed with BB-12® 0 5.36 ± 0.00 ns 0.418 ± 0.00 c 52.89 ± 0.15 a 11.55 ± 0.87 a 3 5.38 ± 0.10 ns 0.425 ± 0.00 b 51.09 ± 0.65 b 10.01 ± 0.34 b 6 5.42 ± 0.08 ns 0.435 ± 0.00 a 50.12 ± 0.34 c 9.12 ± 0.14 c Aw, water activity; WSI, water solubility index; WAI, water absorption index Data are presented as mean ± SD of three measurements (n = 3). Means in columns with superscript letters indicate significant differences ( p < 0.05). However, the determination of bioactive compounds and bioactivities—such as GABA, TPC, antioxidant activity, and AChE and BChE inhibition—was not detectable in any of the germinated Riceberry beverages (data not shown). This may be due to the relatively low concentration of bioactive compounds (approximately 29%) in the germinated Riceberry powder used for developing the probiotic powder and beverage. 3.9 Viable probiotic of geminated Riceberry powder during storage The viable probiotic counts of germinated Riceberry powder, prepared by mixing cereal-based powder with L. casei 431® and BB-12® powders at mixing ratios of 3% and 5% (w/w) and stored at room temperature for 6 months, are shown in Table 8 . Beverages were prepared using warm water at 80°C prior to microbiological analysis for LAB counts. The results indicated that the growth patterns of L. casei 431® and BB-12® were not significantly different ( p ≥ 0.05). The viable counts of probiotics in the beverages ranged from 6.30 to 6.34 log cfu/mL for L. casei 431® and from 6.35 to 6.37 log 10 /mL for BB-12® during 6 months of storage. Interestingly, the viable counts remained at sufficient levels (> 6 log 10 /mL), meeting the minimum requirement for probiotic cell concentration in foods claiming probiotic functionality [ 3 ]. Table 8 Viable probiotic of geminated Riceberry powder during storage Probiotic powder Storage period (months) LAB (log 10 /mL) Germinated Riceberry-based powder mixed with L. casei 431® 0 6.31 3 6.34 6 6.30 Germinated Riceberry-based powder mixed with BB-12® 0 6.36 3 6.37 6 6.35 LAB, lactic acid bacteria 3.10 Sensory evaluation of probiotic geminated Riceberry beverage The average sensory scores of probiotic-germinated Riceberry beverages are shown in Fig. 2 . Significant differences ( p < 0.05) were observed among the samples for all sensory attributes, including colour, aroma, flavour, viscosity, and overall liking. The average scores ranged from 6.41 to 7.16 for colour, 5.74 to 6.51 for aroma, and 5.89 to 6.65 for flavour. The mean viscosity scores varied between 6.17 and 6.96. Panelists gave the highest overall-liking score (6.65) to the beverage prepared from germinated Riceberry with L. casei 431® at a mixing ratio of 3%, while the lowest score (5.58) was recorded for the beverage containing L. casei 431® at a 5% mixing ratio. This variation in sensory acceptance may be attributed to the distinctive characteristics imparted by the pre-mixed probiotic powder (38). Probiotic supplementation has been reported to influence the aroma of yoghurt by increasing the synthesis of acetic acid and other organic compounds, including acetoin, 2-butanone, and 2-ethyl-1-hexanol [ 39 ]. 4 Conclusion A probiotic-germinated pigmented rice beverage was successfully developed using Bifidobacterium animalis subsp. lactis (BB-12®) and Lactobacillus paracasei (L. casei 431®). Levels of GABA, TPC, antioxidant activity, and AChE and BChE inhibitory activities increased as Riceberry germination progressed. Germinated Riceberry powder fortified with 3% and 5% pre-mixed freeze-dried probiotic powders achieved viable probiotic counts above 6 log cfu/mL, meeting the recommended threshold for probiotic efficacy. Beverages prepared with warm water at 80°C retained higher viable probiotic counts. The probiotic-germinated Riceberry drinks also exhibited favourable sensory attributes, particularly in colour, aroma, and flavour. Overall-liking scores indicated moderate acceptance (“like moderately”) for beverages containing BB-12® (3% and 5%) and L. casei 431® (3%). Furthermore, probiotic-germinated Riceberry powder packaged in vacuum-sealed aluminium pouches and stored at room temperature remained shelf-stable, maintaining high probiotic viability and good quality for up to 6 months. Declarations Acknowledgements The authors acknowledge the support from Mae Fah Luang University - the research and innovation strategies under the national strategy research framework. Funding declaration This project received the financial support from Mae Fah Luang University research fund grant contact No. CCF_000077 /2566. Declarations conflict of interest There are no conflicts to declare. Ethics approval and consent to participate Ethical approval for this study was obtained from the Mae Fah Luang University Ethical Committee for conducting the sensory evaluation using human subjects in accordance with the he Ethical Guidelines for Research on Human Subjects in Thailand, B.E. 2550 and adhered to the principles outlined in the Helsinki Declaration. Informed consent was obtained from the panelist for the sensory analysis. Consent for publication Not Applicable. 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1","display":"","copyAsset":false,"role":"figure","size":97345,"visible":true,"origin":"","legend":"\u003cp\u003eGABA content in germinated rice\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8117321/v1/dda02cd090324fb4eccb7e87.png"},{"id":98172374,"identity":"25b71a2c-3c8f-424b-992d-ca7a8547ab52","added_by":"auto","created_at":"2025-12-14 17:33:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":163316,"visible":true,"origin":"","legend":"\u003cp\u003eAverage scores of probiotic geminated Riceberry beverage\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8117321/v1/eaea13b19c1589d1bddc3a85.png"},{"id":100592354,"identity":"9d7d86d1-1434-4baa-b775-9c728782ee4a","added_by":"auto","created_at":"2026-01-19 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Intentionally modified functional foods, including the probiotication of foods\u0026mdash;enriched with probiotics such as lactic acid bacteria (LAB) in adequate amounts\u0026mdash;aim to provide scientifically proven benefits to support gut health [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Probiotics are living microorganisms that, when administered in sufficient quantities, confer a health benefit on the host. According to World Health Organization/Food and Agriculture Organization recommendations, any food claiming a probiotic effect must contain at least 6 log cfu/mL to 7 log cfu/mL (10\u003csup\u003e6\u003c/sup\u003e to 10\u003csup\u003e7\u003c/sup\u003e cfu/mL) of viable probiotic bacteria [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. LAB, including members of the \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eBifidobacterium\u003c/em\u003e genera, exhibit probiotic properties. These genera are part of the indigenous microbiota of humans and animals and are generally recognised as safe [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Consequently, there has been an expansion in the application of probiotics not only in nutraceuticals but also as functional ingredients in foods [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Several studies have reported that cereal-based ingredients\u0026mdash;such as maize, millet, barley, oat, rye, wheat, and rice\u0026mdash;fermented with probiotics can enhance consumer health by combining the benefits of probiotics with health-promoting bioactive components [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRice (\u003cem\u003eOryza sativa\u003c/em\u003e L.) is a staple food for most of the global population because of its nutritional value, quality characteristics, and diverse varieties that meet consumer demand. In 2023, the global harvested rice area was 168,356,566 hectares, while total rice production reached 799,999,504.87 tonnes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. White or polished rice is widely consumed, whereas pigmented varieties such as black, red, and purple rice are preferred by certain consumer groups. Pigmented rice accumulates bioactive metabolites, including cyanidin-3-glucoside, cyanidin-3-galactoside, malvidin-3-galactoside, and peonidin-3-glucoside. In addition, bioactive compounds in pigmented rice offer potential health benefits, including antioxidant, anti-inflammatory, anti-cancer, and anti-diabetic activities [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. In Thailand, a purple rice cultivar known as Riceberry\u0026mdash;a novel variety rich in antioxidant properties and other bioactive compounds\u0026mdash;is available to consumers [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Furthermore, bioactive compounds such as gamma-aminobutyric acid (GABA) in cereal grains can be enhanced through germination, a process influenced by factors such as temperature, time, pH, processing method, and drying conditions that affect biochemical reactions within plant cells. Germination increases the bioavailability of nutrients such as carbohydrates, proteins, GABA, and antioxidants, while reducing anti-nutritional factors such as phytates [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Moreover, germinated rice exhibits improved sensory qualities, including sweetness, texture, and ease of cooking. Therefore, germination plays a crucial role in enhancing the nutritional value and overall quality of brown rice [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Consequently, germinated rice grains can be utilised as a pre-prepared ingredient for developing value-added products such as rice flour, rice milk, rice starch, rice puddings, and rice cakes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003ePowdered food ingredients, such as spray-dried and freeze-dried powders, are widely produced by the food industry because they are easier to process, handle, transport, and store. In addition, instant food powders offer consumers convenient, minimal-preparation options and can be formulated with functional ingredients [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Various types of instant foods\u0026mdash;such as instant germinated purple rice tea [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] and black rice-based instant beverage mixes [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u0026mdash;have been developed using healthy or functional ingredients. Moreover, the enrichment of instant food powders with LAB strains has been explored. Lipan et al [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] developed spray-dried almond milk powder containing \u003cem\u003eLactobacillus plantarum\u003c/em\u003e (ATCC 8014) at concentrations above 10\u003csup\u003e7\u003c/sup\u003e viable cells, maintaining potential probiotic functionality for up to 8 months when stored at 4\u0026deg;C and 6 months at 22\u0026deg;C. However, spray drying at high temperatures remains disadvantageous for drying bioactive or heat-sensitive compounds such as enzymes, cells, and vitamins [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Additionally, designing non-dairy instant powders, such as cereal-based powders, can be challenging because of their macromolecular composition, structural complexity, and the high degree of formulation flexibility required. Rittisak et al. [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] developed an instant beverage using broken Riceberry rice through the application of a double drum dryer operated at 125\u0026deg;C and a drum speed of 1.0 rpm. The resulting dried powder exhibited a light purple colour, low water activity (0.354), and minimal microbial contamination, with total plate counts and yeast and mould levels below 10 cfu/g.\u003c/p\u003e\u003cp\u003eThe formulation and process for enriching plant-based products with phenolic compounds, dietary fibres, and/or probiotics should be carefully tailored to achieve the desired quality characteristics and improve viable probiotic counts [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] Because instant powdered beverages typically require rehydration before consumption, their reconstitution ability\u0026mdash;along with the retention of nutrients, functional compounds, and associated health benefits\u0026mdash;is a crucial techno-functional property for both production and storage [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eDespite growing interest in functional beverages, the development of probiotic cereal drinks and the effects of probiotic enrichment on viable cell counts have not been extensively studied. Furthermore, the existing literature offers limited information in this area, particularly regarding the development of beverages using germinated pigmented rice as an ingredient combined with novel probiotication techniques. Therefore, the objectives of this study were to develop a novel probiotic beverage from germinated pigmented rice powder and to evaluate the viability of probiotics such as \u003cem\u003eBifidobacterium animalis\u003c/em\u003e subsp. lactis (BB-12\u0026reg;) and \u003cem\u003eLactobacillus paracasei\u003c/em\u003e (L. casei 431\u0026reg;) following beverage preparation and storage.\u003c/p\u003e"},{"header":"2 Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Plant materials\u003c/h2\u003e\u003cp\u003eRice (\u003cem\u003eOryza sativa\u003c/em\u003e L.) cultivars Hom Mali 105, RD6, Khiao Ngu, and Riceberry were obtained from a local market in Chiang Rai Province, Thailand.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e2.2 Lactic acid cultures\u003c/b\u003e\u003c/h2\u003e\u003cp\u003eLactic acid-forming bacteria, including \u003cem\u003eBifidobacterium animalis\u003c/em\u003e subsp. lactis (BB-12\u0026reg;) and \u003cem\u003eLactobacillus paracasei\u003c/em\u003e (L. casei 431\u0026reg;), were obtained as Direct Vat Set cultures (10-pouch, 50 U size) from Chr. Hansen, Hoersholm, Denmark.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e2.3 Chemicals\u003c/b\u003e\u003c/h2\u003e\u003cp\u003eAll chemicals used in this study were of analytical reagent grade, mainly manufactured by Sigma-Aldrich (Chemie GmbH, Germany) and supplied by local chemical distributors in Thailand. Purified water was produced using a Milli-Q purification system (Merck Millipore, Bedford, MA, USA).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Germination\u003c/h2\u003e\u003cp\u003eTwenty grams of paddy rice were washed with tap water and soaked in distilled water for 12 hours. After soaking, the samples were rinsed three times with distilled water and drained. The soaked rice grains were then placed in germination boxes lined with clean, sterile cloth and kept in a dark place at 30\u0026deg;C for 24, 48, 72, 96, and 120 hours. At each designated germination time, the samples were washed with distilled water and freeze-dried. The dried samples were ground into a 60-mesh powder and stored in sealed plastic bags at 4\u0026deg;C for further analysis of bioactive compounds and bioactivities.\u003c/p\u003e\u003cp\u003eThe active compounds and bioactivities\u0026mdash;including GABA, total phenolic compounds, antioxidant activity, and cholinesterase enzyme inhibition\u0026mdash;were evaluated in the germinated rice and used as selection criteria for developing the probiotic rice beverage.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Production of germinated rice-based flake\u003c/h2\u003e\u003cp\u003eThe production of the cereal flake was carried out based on a preliminary formulation (data not shown). The optimised recipe consisted of germinated Riceberry flour (29.4%) combined with dry ingredients, including inulin (20.0%), whey protein (15.0%), maltodextrin (10.0%), corn-based milk powder alternative (10.0%), full-cream milk powder (5.0%), malt powder (5.0%), and sucralose (0.1%). The dry mixture was then blended with reverse osmosis water at a mixing ratio of 1:2 (w/w). Xanthan gum (0.5%) was subsequently added. The mixture was homogenised using a KitchenAid stand mixer (Artisan\u0026reg;, USA) and then dried using a twin drum dryer (Euro Best Technology Co., Ltd., Bangkok, Thailand) at 150\u0026deg;C with a roller rotation speed of 1 rpm. The resulting germinated Riceberry flake was collected and cooled to room temperature. The cereal flakes were then ground in a cooling-assisted grinder and stored at room temperature for subsequent development into probiotic-germinated Riceberry powder.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.6 Production of freeze-dried probiotic powder\u003c/h2\u003e\u003cp\u003eProbiotic cultures, including BB-12\u0026reg; and L. casei 431\u0026reg;, were inoculated to achieve an initial viable bacterial level of at least 6 log cfu/g. The liquid milk was pasteurised at 73\u0026deg;C for 15 seconds and then cooled to 43\u0026deg;C\u0026ndash;45\u0026deg;C. The commercial freeze-dried starter was subsequently added and mixed thoroughly for 10\u0026ndash;15 minutes. Fermentation was carried out at 45\u0026deg;C for 24 hours. After fermentation, the yoghurt was freeze-dried at a shelf temperature of 20\u0026deg;C using a laboratory-scale freeze dryer (Martin Christ Gefriertrocknungsanlagen GmbH, Germany). The resulting lyophilised flakes were ground in a cooling-assisted grinder and stored at room temperature before being mixed with germinated Riceberry powder.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e2.7 Production of probiotic-germinated rice powder\u003c/h2\u003e\u003cp\u003eGerminated Riceberry powder (see Section \u003cspan refid=\"Sec7\" class=\"InternalRef\"\u003e2.5\u003c/span\u003e) was prepared and fortified with freeze-dried BB-12\u0026reg; and L. casei 431\u0026reg; powders (see Section \u003cspan refid=\"Sec8\" class=\"InternalRef\"\u003e2.6\u003c/span\u003e) at mixing ratios of 3% (w/w) and 5% (w/w), respectively. A 30-g portion of the probiotic cereal powder was packed in a laminated aluminium pouch with a low-density polyethylene (LDPE/AL/LDPE) lining (10- \u0026times; 15-cm size, 110-\u0026micro;m thickness). The shelf-life study of the samples was conducted at 35\u0026deg;C for a storage period of 6 months.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e2.8 Determination of GABA content in germinated rice\u003c/h2\u003e\u003cp\u003eThe analysis of GABA content was carried out following the method of Kamjijam et al [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] with slight modifications. A 0.5-g sample was placed in a test tube and mixed with 0.9 mL of distilled water and 0.9 mL of ethanol containing 3% sulfosalicylic acid (v/v). The solution was stirred thoroughly and centrifuged at 5,000 rpm for 10 minutes. An aliquot of the supernatant (50 \u0026micro;L) was then mixed with 50 \u0026micro;L of NaHCO\u003csub\u003e3\u003c/sub\u003e solution and 200 \u0026micro;L of DABSYL-Cl acetonitrile solution. The mixture was incubated at 70\u0026deg;C for 10 minutes, followed by the addition of 250 \u0026micro;L of ethanol and 250 \u0026micro;L of KH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e buffer solution (pH 6.8). The mixture was then centrifuged at 4,500 rpm for 10 minutes and filtered through a 0.45-\u0026micro;m nylon membrane filter. The resulting solution was analysed using an Acquity\u0026trade; UPLC system (Waters Acquity\u0026trade;; Waters Corp., Milford, MA, USA) equipped with a BEH C18 column (100 mm \u0026times; 2.1 mm, 1.7 \u0026micro;m; Waters Corp.) and a UV detector set at 465 nm.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e2.9 Determination of total phenolic content (TPC)\u003c/h2\u003e\u003cp\u003eThe TPC was determined using the Folin\u0026ndash;Ciocalteu assay [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. An aliquot of the germinated rice extract (20 \u0026micro;L) was mixed with 100 \u0026micro;L of 10% Folin\u0026ndash;Ciocalteu reagent and 80 \u0026micro;L of 7.5% sodium carbonate solution. The mixture was incubated in the dark at room temperature for 30 minutes. The absorbance was then measured at 760 nm using a UV\u0026ndash;visible spectrophotometer (Biochrom Libra S22, UK). TPC was calculated using a gallic acid standard curve, and the results were expressed as milligrams of gallic acid equivalents per 100 g dry matter (mg GAE/100 g DM).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e2.10 Determination of antioxidant activity\u003c/h2\u003e\u003cp\u003eThe antioxidant activity was determined using the DPPH radical scavenging assay, following the method described by Netrprachit et al. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] with slight modifications. An extract (50 \u0026micro;L) was mixed with 100 \u0026micro;L of DPPH solution in methanol (5.9 mg/100 mL, w/v) in a 96-well plate. The mixture was then incubated in the dark for 30 minutes. The absorbance was measured at 517 nm using a 96-well plate UV\u0026ndash;Vis spectrophotometer (Thermo Fisher Scientific Inc., MA, USA). The control consisted of distilled water (250 \u0026micro;L) in place of the extract. The percentage of antioxidant activity was then calculated by comparing the absorbance of the sample to that of the control.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e2.11 Determination of inhibitory potential against cholinesterase enzymes\u003c/h2\u003e\u003cp\u003eThe determination of acetylcholinesterase (AChE) inhibition in the germinated rice extract was carried out following the method of Temviriyanukul et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] with slight modifications. An extract (50 \u0026micro;L) was mixed with an enzyme solution consisting of \u003cem\u003eElectrophorus electricus\u003c/em\u003e AChE (20 ng; 1000 units/mg, 100 \u0026micro;L) in 50 mM potassium phosphate buffer (KPB, pH 7.0) in a 96-well plate. Subsequently, 10 \u0026micro;L of 16 mM 5,5\u0026prime;-dithio-bis-(2-nitrobenzoic acid) (DTNB) and 40 \u0026micro;L of 0.8 mM acetylthiocholine in 50 mM KPB (pH 7.0) were added to the mixture. The absorbance was measured at 412 nm using a 96-well plate UV\u0026ndash;Vis spectrophotometer (Thermo Fisher Scientific Inc.). The percentage of inhibition was then calculated.\u003c/p\u003e\u003cp\u003eThe determination of butyrylcholinesterase (BChE) inhibition was performed using the same procedure. A solution containing 100 ng of equine serum BChE (\u0026ge;\u0026thinsp;10 units/mg protein, 100 \u0026micro;L) in 50 mM KPB (pH 7.0) with 1 mM MgCl\u003csub\u003e2\u003c/sub\u003e and 0.1 mM butyrylthiocholine (BTCh) in 50 mM KPB (pH 7.0) served as the reactive enzyme\u0026ndash;substrate mixture. The absorbance was recorded at 412 nm using a 96-well plate UV\u0026ndash;Vis spectrophotometer (Thermo Fisher Scientific Inc.), and the percentage of inhibition was calculated accordingly.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e2.12 Enumeration of probiotic cultures\u003c/h2\u003e\u003cp\u003eEnumeration of LAB was performed using the 3M Petrifilm\u0026reg; Lactic Acid Bacteria Count Plate method [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] (3M Inc., MN, USA; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.3m.com\u003c/span\u003e\u003cspan address=\"https://www.3m.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). The number of colonies was expressed as log\u003csub\u003e10\u003c/sub\u003e cfu/g.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e2.13 Enumeration of viable probiotic in geminated Riceberry beverage\u003c/b\u003e\u003c/h2\u003e\u003cp\u003eA 15-g sample of probiotic-germinated Riceberry powder was dissolved in 150 mL of warm water at 60\u0026deg;C, 80\u0026deg;C, and 100\u0026deg;C, with stirring for 3 minutes to ensure complete dissolution. The prepared beverages were then transferred into 50-mL sealed plastic tubes and stored at \u0026minus;\u0026thinsp;20\u0026deg;C for subsequent microbial colony analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e2.14 Enumeration of viable probiotic in geminated rice beverage during storage\u003c/h2\u003e\u003cp\u003eSamples were collected every 30 days during storage to determine the viable probiotic count, [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] as described in Section \u003cspan refid=\"Sec15\" class=\"InternalRef\"\u003e2.13\u003c/span\u003e.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e2.15 Determination physicochemical property of geminated rice beverage during storage\u003c/h2\u003e\u003cp\u003eWater activity (Aw) was measured using an Aw meter (AQUALAB Model Series 3TE; AquaLab, Pullman, WA, USA). Moisture content was determined according to the AOAC method 925.10, [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] as established by the Association of Official Analytical Chemists. In addition, the water solubility index (WSI) and water absorption index (WAI) were determined following the method of Kim et al. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e2.16 Sensory evaluation of probiotic-germinated rice beverage\u003c/h2\u003e\u003cp\u003eSensory evaluation was conducted with 30 trained panellists (aged 30\u0026ndash;45 years), recruited from students and staff of the School of Agro-Industry, Mae Fah Luang University. A 15-g sample of probiotic-germinated rice powder was dissolved in 150 mL of warm water at 80\u0026deg;C and stirred for 3 minutes to ensure complete dissolution. The test samples were served in 15-mL glass cups at room temperature. Panellists evaluated and scored the sensory attributes\u0026mdash;appearance, colour, odour, taste, viscosity, and overall liking\u0026mdash;using a 9-point hedonic scale [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] (1\u0026thinsp;=\u0026thinsp;dislike very much, 9\u0026thinsp;=\u0026thinsp;like very much).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\u003ch2\u003e2.17 Statistical analysis\u003c/h2\u003e\u003cp\u003eAll data were statistically analysed using analysis of variance, and mean comparisons were performed using Duncan\u0026rsquo;s multiple range test to determine significant differences between samples at a 95% confidence level (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Statistical analyses were carried out using IBM SPSS\u0026reg; Statistics version 21.0.0 (IBM Corp., Armonk, NY, USA).\u003c/p\u003e\u003c/div\u003e"},{"header":"3 Results and Discussion","content":"\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\u003ch2\u003e3.1 GABA content in germinated rice\u003c/h2\u003e\u003cp\u003eThe germination of brown rice can improve its flavour, enhance its nutritional value, and increase its potential health benefits. Germinated brown rice is richer in nutrients and bioactive compounds such as essential amino acids, phenolic acids, γ-oryzanol, and GABA [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Therefore, germinated brown rice is increasingly recognised as a promising health food. In this study, germination was performed on four rice varieties: Hom Mali 105, Riceberry, glutinous RD6, and Khiao Ngu. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the GABA content in the germinated rice samples changed significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) during the 120-hour germination period. Non-waxy rice varieties, including Hom Mali 105 and Riceberry, exhibited higher GABA levels than did waxy varieties such as RD6 and Khiao Ngu. The GABA content ranged from 4.78 to 16.41 g/100 g DM for Hom Mali 105 and from 7.93 to 26.01 g/100 g DM for Riceberry. The maximum GABA accumulation occurred between 48 and 72 hours of germination for both Hom Mali 105 and Riceberry, after which levels plateaued or declined. Chaijan and Panpipat [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] reported that the optimal germination time for indigenous Southern Thai rice varieties, including Khemtong and Khai Mod Rin, was 96 hours, resulting in the highest levels of GABA, thiamine, free amino acids, total sugars, and α-amylase activity. The increase in GABA content is likely due to the activity of the enzyme glutamate decarboxylase, which catalyses the conversion of glutamate to GABA [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Similarly, C\u0026aacute;ceres et al. [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] reported that GABA synthesis occurs in a time-dependent manner during germination.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\u003ch2\u003e3.2 TPC of germinated rice\u003c/h2\u003e\u003cp\u003eLonger germination periods resulted in an increase in the TPC of the rice samples (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). A significant difference (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in TPC was observed at all 24-hour intervals during germination. Germinated Riceberry showed the highest TPC, increasing from 163.43 mg GAE/100 g DW at 0 h to 299.53 mg GAE/100 g DW at 120 h. Similarly, Singh et al. (2017) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] reported that germination of non-waxy Thai rice cultivar Phitsanulok 2 at 33\u0026deg;C for 300 minutes increased both GABA and TPC levels\u0026mdash;from 6.71 mg/100 g and 13.68 mg GAE/100 g DM to 18.74 mg/100 g and 33.95 mg GAE/100 g DM, respectively. Structural changes in the grain during germination and mashing are related to various enzymatic degradations that facilitate the release and extraction of free polyphenols [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] Enzyme activities may also vary considerably depending on temperature and pH conditions.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eTotal phenolic content of germinated rice\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eGerminated rice extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u003cp\u003eTotal phonic content (mg GAE/100 g DW)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e72 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e96 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e120 h\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHom Mali 105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e65.31\u0026thinsp;\u0026plusmn;\u0026thinsp;5.31 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e74.03\u0026thinsp;\u0026plusmn;\u0026thinsp;3.35 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e88.16\u0026thinsp;\u0026plusmn;\u0026thinsp;6.43 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e110.13\u0026thinsp;\u0026plusmn;\u0026thinsp;5.20 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e98.15\u0026thinsp;\u0026plusmn;\u0026thinsp;4.59 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e109.83\u0026thinsp;\u0026plusmn;\u0026thinsp;5.77 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRiceberry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e163.43\u0026thinsp;\u0026plusmn;\u0026thinsp;3.53 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e170.11\u0026thinsp;\u0026plusmn;\u0026thinsp;4.67 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e184.74\u0026thinsp;\u0026plusmn;\u0026thinsp;8.67 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e200.84\u0026thinsp;\u0026plusmn;\u0026thinsp;3.42 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e257.91\u0026thinsp;\u0026plusmn;\u0026thinsp;3.45 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e299.53\u0026thinsp;\u0026plusmn;\u0026thinsp;5.77 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRD6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e43.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e47.71\u0026thinsp;\u0026plusmn;\u0026thinsp;2.71 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e53.08\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e55.29\u0026thinsp;\u0026plusmn;\u0026thinsp;3.52 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e58.34\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKhiao Ngu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e16.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e17.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e14.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e20.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e17.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e19.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 \u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD of three measurements (n\u0026thinsp;=\u0026thinsp;3).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eMeans in columns with superscript letters indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\u003ch2\u003e3.3 Antioxidant activity of germinated rice\u003c/h2\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e presents the antioxidant activity of 50 \u0026micro;g/mL extracts from both ungerminated and germinated rice samples. The initial antioxidant activity of the ungerminated rice samples showed significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Among them, the Riceberry extract exhibited the highest percentage of DPPH inhibition (35.07% \u0026plusmn; 0.89%). The antioxidant activity of non-pigmented rice varieties was comparatively lower than that of pigmented rice. Black rice varieties are rich in anthocyanins, primarily cyanidin-3-glucoside and peonidin-3-glucoside, which range from 19.4 to 140.8 mg/100 g DM and 11.1 to 12.8 mg/100 g DM, respectively [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The biological functionality of pigmented rice is largely attributed to its polyphenolic compounds. Furthermore, the antioxidant activity of all germinated extracts increased significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with longer germination periods, indicating that germination positively affects antioxidant properties. Several studies have reported similar findings, showing that the TPC of germinated black waxy rice is higher than that of non-germinated rice, while germinated red jasmine rice exhibits slightly lower values. Phattayakorn et al. [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] also reported that germinated rice demonstrated higher antioxidant activity than non-germinated rice, as determined by FRAP and DPPH assays.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAntioxidant activity (DPPH scavenging) of germinated rice\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eGerminated rice extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u003cp\u003eAntioxidant potential (%) of 50 \u0026micro;g/mL germinated rice extract\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e72 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e96 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e120 h\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHom Mali 105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e9.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e9.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e12.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e14.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRiceberry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e35.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e37.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.91 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e47.57\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e56.31\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e65.46\u0026thinsp;\u0026plusmn;\u0026thinsp;2.51 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e71.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRD6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e9.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28 \u003csup\u003eb,c\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.85 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e10.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.92 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKhiao Ngu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e8.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e10.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD of three measurements (n\u0026thinsp;=\u0026thinsp;3).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eMeans in columns with superscript letters indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\u003ch2\u003e3.4 AChE and BChE inhibitory potential of germinated rice\u003c/h2\u003e\u003cp\u003eThe extracts of germinated rice demonstrated inhibitory potential against cholinesterase enzyme activities (Tables\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Germinated pigmented rice cultivar Riceberry showed higher AChE and BChE inhibitory activities than germinated non-pigmented rice at all germination periods. In comparison, the extract of parboiled germinated brown rice exhibited 6% AChE inhibition and 39% BChE inhibition, with significantly higher inhibitory activity than other rice types, including germinated brown rice, brown rice, and white rice [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. The enhanced inhibitory effects may be attributed to bioactive peptides generated during rice germination and mashing, which can contribute to angiotensin-converting enzyme, AChE, and BChE inhibitory capacities [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAChE inhibitory potential of germinated rice\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eGerminated rice extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u003cp\u003eAChE inhibitory potential of 50 \u0026micro;g/mL germinated rice extract\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e72 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e96 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e120 h\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHom Mali 105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e8.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e14.71\u0026thinsp;\u0026plusmn;\u0026thinsp;3.7 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e17.86\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e18.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRiceberry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e26.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e26.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e24.71\u0026thinsp;\u0026plusmn;\u0026thinsp;3.77 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e26.86\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e28.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRD6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.91\u0026thinsp;\u0026plusmn;\u0026thinsp;1.22 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.49\u0026thinsp;\u0026plusmn;\u0026thinsp;1.45 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e3.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKhiao Ngu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD of three measurements (n\u0026thinsp;=\u0026thinsp;3).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eMeans in columns with superscript letters indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBChE inhibitory potential of germinated rice\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eGerminated rice extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c7\" namest=\"c2\"\u003e\u003cp\u003eBChE inhibitory potential of 50 \u0026micro;g/mL germinated rice sample extract\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e48 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e72 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e96 h\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e120 h\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHom Mali 105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e35.19\u0026thinsp;\u0026plusmn;\u0026thinsp;1.50 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e33.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.78 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e37.81\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e40.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e41.09\u0026thinsp;\u0026plusmn;\u0026thinsp;1.92 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e41.98\u0026thinsp;\u0026plusmn;\u0026thinsp;2.45 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRiceberry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e41.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e43.91\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e51.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e55.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e59.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e59.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRD6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e26.43\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e30.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.57 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e28.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e30.32\u0026thinsp;\u0026plusmn;\u0026thinsp;1.23 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e31.53\u0026thinsp;\u0026plusmn;\u0026thinsp;1.99 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKhiao Ngu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e19.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e20.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e21.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e22.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.78 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e21.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD of three measurements (n\u0026thinsp;=\u0026thinsp;3).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eMeans in columns with superscript letters indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e\u003ch2\u003e3.5 Colony counts of probiotic starter in pre-mixed powder\u003c/h2\u003e\u003cp\u003eThe colony counts of probiotic starters L. casei 431\u0026reg; and BB-12\u0026reg; in the pre-mixed powder were 8.95 and 8.92 log\u003csub\u003e10\u003c/sub\u003e cfu/g, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Freeze-drying effectively preserves probiotic viability, maintaining the strains\u0026rsquo; original chemical composition and physical properties, while enabling long-term storage and good fermentation performance [36\u0026thinsp;\u0026minus;\u0026thinsp;7].\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eColony counts of probiotic starter in pre-mixed powder\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"2\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStarter\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLAB (log\u003csub\u003e10\u003c/sub\u003e/g)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. paracasei\u003c/em\u003e subs. paracasei, L. casei 431\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e8.95\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eB. animalis\u003c/em\u003e subsp. lactis, BB-12\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e8.92\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"2\"\u003eLAB, lactic acid bacteria\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e\u003ch2\u003e3.6 Production of probiotic-germinated Riceberry powder\u003c/h2\u003e\u003cp\u003eGermination of Riceberry for 120 hours resulted in the highest levels of GABA, TPC, antioxidant activity, and AChE and BChE inhibitory potentials (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Therefore, germinated Riceberry was selected as the raw material for preparing germinated rice-based flakes (Section \u003cspan refid=\"Sec7\" class=\"InternalRef\"\u003e2.5\u003c/span\u003e) and subsequently used to produce probiotic-germinated Riceberry powder (Section \u003cspan refid=\"Sec9\" class=\"InternalRef\"\u003e2.7\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec27\" class=\"Section2\"\u003e\u003ch2\u003e3.7 Viable probiotic counts in geminated Riceberry beverage\u003c/h2\u003e\u003cp\u003eBeverages prepared from germinated Riceberry-based powder mixed with L. casei 431\u0026reg; and BB-12\u0026reg; at mixing ratios of 3% and 5% (w/w) were reconstituted with warm water at 60\u0026deg;C, 80\u0026deg;C, and 100\u0026deg;C and analysed for LAB counts (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The results showed that the viability of both probiotic strains decreased slightly at both mixing ratios. Increasing the reconstitution temperature led to a reduction in viable counts of L. casei 431\u0026reg; and BB-12\u0026reg; in the probiotic beverages. The final counts decreased by 0.17\u0026ndash;0.21 log\u003csub\u003e10\u003c/sub\u003e/mL for L. casei 431\u0026reg; and 0.14\u0026ndash;0.24 log cfu/mL for BB-12\u0026reg;. The percentage of reduction ranged from 31.38% to 58.68% for L. casei 431\u0026reg; and BB-12\u0026reg;, respectively, when beverages were prepared with warm water at 60\u0026deg;C\u0026ndash;80\u0026deg;C. Notably, probiotic beverages prepared at 60\u0026deg;C and 80\u0026deg;C still retained viable counts above 6 log\u003csub\u003e10\u003c/sub\u003e/mL for both strains. This indicates that the viable counts of L. casei 431\u0026reg; and BB-12\u0026reg; in the novel germinated Riceberry beverages prepared at 60\u0026deg;C and 80\u0026deg;C met the minimum threshold of 6\u0026ndash;7 log cfu/mL required for foods claiming probiotic functionality [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eViable probiotic counts in geminated Riceberry beverage\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eProbiotic beverage powder\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMixing ratio\u003c/p\u003e\u003cp\u003e(% w/w)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWarm water temperature\u003c/p\u003e\u003cp\u003e(\u0026deg;C)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLAB\u003c/p\u003e\u003cp\u003e(log\u003csub\u003e10\u003c/sub\u003e/mL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"5\" rowspan=\"6\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with L. casei 431\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.22\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.84\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.57\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.40\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.23\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"5\" rowspan=\"6\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with BB-12\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.92\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.67\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.43\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.29\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eLAB, lactic acid bacteria\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\u003ch2\u003e3.8 Physicochemical property of probiotic geminated Riceberry powder during storage\u003c/h2\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e presents the physicochemical properties of germinated Riceberry powder during 6 months of storage. Probiotic-germinated Riceberry powder stored under vacuum packaging showed moisture content and water activity values ranging from 5.31% to 5.42% and 0.410 to 0.435, respectively. For reference, intermediate-moisture foods typically have water activity values between 0.60 and 0.85 and moisture contents ranging from 15% to 40%, which provide stability against microbial contamination and lipid oxidation [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. In this study, water activity in the vacuum-packaged samples did not differ significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026ge;\u0026thinsp;0.05) throughout storage. Although water activity increased slightly with longer storage duration, it remained below 0.6, indicating that the powder was stable against undesirable chemical reactions and microbial growth. Furthermore, storage duration influenced the WSI and WAI of the probiotic-germinated Riceberry powder. The powder exhibited high solubility, with WSI values ranging from 53.09% to 49.65% for L. casei 431\u0026reg; and from 52.89% to 50.12% for BB-12\u0026reg;, both of which were largely retained during storage. The WAI of the probiotic powder gradually decreased during storage and was considerably low (29%) in the final month of the storage period.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePhysicochemical properties of geminated Riceberry powder during storage\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eProbiotic powder\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eStorage period (months)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMoisture content (%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAw (-)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eWSI\u003c/p\u003e\u003cp\u003e(%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eWAI\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with L. casei 431\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.410\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e53.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e11.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.416\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e51.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.430\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e49.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with BB-12\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.418\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e52.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e11.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.425\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e51.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 \u003csup\u003ens\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.435\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e50.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14 \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eAw, water activity; WSI, water solubility index; WAI, water absorption index\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eData are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD of three measurements (n\u0026thinsp;=\u0026thinsp;3).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eMeans in columns with superscript letters indicate significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eHowever, the determination of bioactive compounds and bioactivities\u0026mdash;such as GABA, TPC, antioxidant activity, and AChE and BChE inhibition\u0026mdash;was not detectable in any of the germinated Riceberry beverages (data not shown). This may be due to the relatively low concentration of bioactive compounds (approximately 29%) in the germinated Riceberry powder used for developing the probiotic powder and beverage.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec29\" class=\"Section2\"\u003e\u003ch2\u003e3.9 Viable probiotic of geminated Riceberry powder during storage\u003c/h2\u003e\u003cp\u003eThe viable probiotic counts of germinated Riceberry powder, prepared by mixing cereal-based powder with L. casei 431\u0026reg; and BB-12\u0026reg; powders at mixing ratios of 3% and 5% (w/w) and stored at room temperature for 6 months, are shown in Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e. Beverages were prepared using warm water at 80\u0026deg;C prior to microbiological analysis for LAB counts. The results indicated that the growth patterns of L. casei 431\u0026reg; and BB-12\u0026reg; were not significantly different (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026ge;\u0026thinsp;0.05). The viable counts of probiotics in the beverages ranged from 6.30 to 6.34 log cfu/mL for L. casei 431\u0026reg; and from 6.35 to 6.37 log\u003csub\u003e10\u003c/sub\u003e/mL for BB-12\u0026reg; during 6 months of storage. Interestingly, the viable counts remained at sufficient levels (\u0026gt;\u0026thinsp;6 log\u003csub\u003e10\u003c/sub\u003e/mL), meeting the minimum requirement for probiotic cell concentration in foods claiming probiotic functionality [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eViable probiotic of geminated Riceberry powder during storage\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eProbiotic powder\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eStorage period (months)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLAB (log\u003csub\u003e10\u003c/sub\u003e/mL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with L. casei 431\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.31\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.34\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.30\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003eGerminated Riceberry-based powder mixed with BB-12\u0026reg;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.36\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.37\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6.35\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003eLAB, lactic acid bacteria\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec30\" class=\"Section2\"\u003e\u003ch2\u003e3.10 Sensory evaluation of probiotic geminated Riceberry beverage\u003c/h2\u003e\u003cp\u003eThe average sensory scores of probiotic-germinated Riceberry beverages are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were observed among the samples for all sensory attributes, including colour, aroma, flavour, viscosity, and overall liking. The average scores ranged from 6.41 to 7.16 for colour, 5.74 to 6.51 for aroma, and 5.89 to 6.65 for flavour. The mean viscosity scores varied between 6.17 and 6.96. Panelists gave the highest overall-liking score (6.65) to the beverage prepared from germinated Riceberry with L. casei 431\u0026reg; at a mixing ratio of 3%, while the lowest score (5.58) was recorded for the beverage containing L. casei 431\u0026reg; at a 5% mixing ratio. This variation in sensory acceptance may be attributed to the distinctive characteristics imparted by the pre-mixed probiotic powder (38). Probiotic supplementation has been reported to influence the aroma of yoghurt by increasing the synthesis of acetic acid and other organic compounds, including acetoin, 2-butanone, and 2-ethyl-1-hexanol [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"4 Conclusion","content":"\u003cp\u003eA probiotic-germinated pigmented rice beverage was successfully developed using \u003cem\u003eBifidobacterium animalis\u003c/em\u003e subsp. lactis (BB-12\u0026reg;) and \u003cem\u003eLactobacillus paracasei\u003c/em\u003e (L. casei 431\u0026reg;). Levels of GABA, TPC, antioxidant activity, and AChE and BChE inhibitory activities increased as Riceberry germination progressed. Germinated Riceberry powder fortified with 3% and 5% pre-mixed freeze-dried probiotic powders achieved viable probiotic counts above 6 log cfu/mL, meeting the recommended threshold for probiotic efficacy. Beverages prepared with warm water at 80\u0026deg;C retained higher viable probiotic counts. The probiotic-germinated Riceberry drinks also exhibited favourable sensory attributes, particularly in colour, aroma, and flavour. Overall-liking scores indicated moderate acceptance (\u0026ldquo;like moderately\u0026rdquo;) for beverages containing BB-12\u0026reg; (3% and 5%) and L. casei 431\u0026reg; (3%). Furthermore, probiotic-germinated Riceberry powder packaged in vacuum-sealed aluminium pouches and stored at room temperature remained shelf-stable, maintaining high probiotic viability and good quality for up to 6 months.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors acknowledge the support from Mae Fah Luang University - the research and innovation strategies under the national strategy research framework.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis project received the financial support from Mae Fah Luang University research fund grant contact No. CCF_000077 /2566.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclarations conflict of interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no conflicts to declare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEthical approval for this study was obtained from the Mae Fah Luang University Ethical Committee for conducting the sensory evaluation using human subjects in accordance with the he Ethical Guidelines for Research on Human Subjects in Thailand, B.E. 2550 and adhered to the principles outlined in the Helsinki Declaration. Informed consent was obtained from the panelist for the sensory analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot Applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHuman Ethics declaration\u003c/strong\u003e:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eFekete M, Lehoczki A, Kryczyk-Poprawa A, Z\u0026aacute;b\u0026oacute; V, Varga JT, B\u0026aacute;lint M, Varga P. Functional foods in modern nutrition science: mechanisms, evidence, and public health implications. 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Technol\u003c/em\u003e\u003cem\u003e.\u003c/em\u003e 2024;17(11): 4354-4370. https://doi.org/10.1007/s11947-024-03405-0.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Pigmented rice, germination, probiotics, bioactive compounds, functional drinks","lastPublishedDoi":"10.21203/rs.3.rs-8117321/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8117321/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eProbiotic cereal-based beverages have gained increasing attention as novel functional and fast-growing products. This study developed a probiotic beverage from germinated pigmented rice-based powder enriched with probiotics, specifically \u003cem\u003eBifidobacterium animalis\u003c/em\u003e subsp. lactis (BB-12\u0026reg;) and \u003cem\u003eLactobacillus paracasei\u003c/em\u003e (L. casei 431\u0026reg;). Indigenous polished and pigmented rice varieties from Thailand were subjected to germination to enhance their bioactive compound content and biological activities. Among the tested varieties, the pigmented rice cultivar Riceberry exhibited the highest concentrations of γ-aminobutyric acid, total phenolic content, antioxidant activity, and acetylcholinesterase and butyrylcholinesterase inhibitory activities. Riceberry flour and other ingredients were used to produce a pigmented cereal-based powder, which was subsequently fortified with freeze-dried probiotic powder at mixing ratios of 3% and 5%. The probiotic pigmented cereal-based powder and its corresponding beverage achieved viable probiotic counts above 6 log cfu/mL, meeting the minimum threshold for probiotic efficacy. The beverage also demonstrated favourable sensory attributes, particularly in colour, aroma, and flavour. Furthermore, probiotic-germinated Riceberry powder packaged in vacuum-sealed aluminium pouches remained shelf-stable under room-temperature storage, maintaining high probiotic viability and good quality for up to 6 months.\u003c/p\u003e","manuscriptTitle":"Production and quality characterizations of probiotic-germinated rice beverage","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-14 17:32:58","doi":"10.21203/rs.3.rs-8117321/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6b2e4355-ea40-45ae-ab94-0bd9694110af","owner":[],"postedDate":"December 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-19T12:57:07+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-14 17:32:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8117321","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8117321","identity":"rs-8117321","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}