Use of pigmented rice as carrier and stingless bee honey as prebiotic to formulate novel synbiotic products mixed with three strains of probiotic bacteria

SWE, Zin Myo; CHUMPHON, Thapakorn; PANGJIT, Kanjana; PROMSAI, Saran

doi:10.1590/fst.120722

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Food Science and Technology

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Original Article • Food Sci. Technol 43 • 2023 • https://doi.org/10.1590/fst.120722 copy

Use of pigmented rice as carrier and stingless bee honey as prebiotic to formulate novel synbiotic products mixed with three strains of probiotic bacteria

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The aim of this study was to produce a synbiotic product containing Thai-pigmented rice as the carrier and the honey of the stingless bee as the prebiotic agent. From antibacterial activity results, all of the tested probiotic bacteria (Limosilactobacillus reuteri KUKPS6103, Lacticaseibacillus rhamnosus KUKPS6007 and Lacticaseibacillus paracasei KUKPS6201) had a high ability to inhibit six strains of intestinal pathogens. All three probiotic bacterial strains had high antioxidant activity according to the ABTS and DPPH scavenging assays. The three tested probiotic bacteria produced substantial reductions in the cholesterol removal percentage of 33.9-78.99%. In the prebiotic studies, stingless bee honey had high enhancing activity and the main carbohydrate components were sucrose, glucose and fructose. A synbiotic product was produced containing Thai-pigmented rice grains (cultivars Riceberry, Tubtim Chumphae and Sangyod) and stingless bee honey. During 8 weeks of storage, the viability of the probiotics in terms of mixed-strains was 7.96 ± 0.06 log CFU g^-1 (90.25% survival rate). Microbiological safety testing stated that the amounts of contaminants were acceptable. This is the first report of an application of Thai-pigmented rice, stingless bee honey and mixed-culture probiotics as a novel functional synbiotic product.

Keywords:
probiotic; prebiotic; synbiotic; pigmented-rice; stingless bee honey

Pathogens	Inhibition zone (mm)
Pathogens	L. reuteri KUKPS6103	L. rhamnosus KUKPS6007	L. paracasei KUKPS6201	Mixed culture	Streptomycin
A. hydrophila	12.33 ± 2.52^a^* * Values represent mean ± standard deviation of three independent experiments. A different English lower case superscript letter in column represents a significantly different value in statistics using Duncan’s test with a confidential level of 95% between each column.	11.67 ± 3.06^a	12.00 ± 1.00^a	11.67 ± 0.56^a	23.33 ± 1.53^b
B. cereus	11.33 ± 0.58^a	12.33 ± 0.58^a	12.00 ± 1.00^a	12.00 ± 0.00^a	22.67 ± 1.53^b
E. coli	11.67 ± 1.53^a	14.00 ± 1.00^a	14.33 ± 1.16^a	13.67 ± 1.16^a	20.67 ± 2.52^b
P. vulgaris	9.33 ± 1.15^a	11.00 ± 1.73^ab	10.00 ± 1.00^a	10.00 ± 1.00^a	16.33 ± 6.66^b
S. typhimurium	11.33 ± 0.58^a	12.67 ± 0.58^a	11.33 ± 0.58^a	11.67 ± 0.58^a	15.67 ± 2.52^b
S. aureus	11.33 ± 0.58^a	11.33 ± 0.58^a	11.00 ± 1.00^a	12.00 ± 1.00^a	25.33 ± 1.15^b

Probiotic bacteria	Antioxidant activity (mg Trolox mL^-1 extract)
L. reuteri KUKPS6103	1.564 ± 0.06^a^* * Values represent mean ± standard deviation of three independent experiments. Different lowercase superscripts represent a significant difference using Duncan’s Test with a confidence level of 95%.
L. rhamnosus KUKPS6007	1.514 ± 0.40^a
L. paracasei KUKPS6201	1.553 ± 0.02^a
Mixed culture	1.516 ± 0.48^a

Probiotic bacteria	Antioxidant activity (mg Gallic acid mL^-1 extract)
L. reuteri KUKPS6103	0.033 ± 0.009^a^* * Values represent mean ± standard deviation of three independent experiments. Different lowercase superscripts represent a significant difference using Duncan’s test with a confidence level of 95%.
L. rhamnosus KUKPS6007	0.096 ± 0.003^b
L. paracasei KUKPS6201	0.084 ± 0.005^b
Mixed culture	0.085 ± 0.005^b

Probiotic bacteria	Honey concentration (g L^-1)
Probiotic bacteria	100	200	300	400	500
L. reuteri KUKPS6103	7.52 ± 0.04^b^* * Values represent mean ± standard deviation of three independent experiments. Different lowercase superscripts represent a significant difference using Duncan’s test with a confidence level of 95%.	7.83 ± 0.07^c	7.83 ± 0.04^c	6.98 ± 0.17^a	7.14 ± 0.09^a
L. rhamnosus KUKPS6007	8.13 ± 0.19^a	7.73 ± 0.40^a	7.94 ± 0.12^a	8.19 ± 0.20^a	7.93 ± 0.07^a
L. paracasei KUKPS6201	7.75 ± 0.04^b	7.40 ± 0.17^a,b	7.79 ± 0.10^b	7.55 ± 0.13^{a, b}	7.16 ± 0.27^a

Probiotic bacteria	Honey (g L^-1)		Inulin (g L^-1)
Probiotic bacteria	20	50	20	50
Enhancing activity (%)
L. reuteri KUKPS6103	3.82	3.44	11.54	4.30
L. rhamnosus KUKPS6007	2.27	0.96	3.46	1.91
L. paracasei KUKPS6201	0.14	4.11	1.58	0.54
Mixed culture	7.39	6.28	9.20	5.19
Enhancing activity (log CFU mL^-1)
L. reuteri KUKPS6103	0.21	0.02	0.20	0.01
L. rhamnosus KUKPS6007	0.15	0.64	0.17	0.04
L. paracasei KUKPS6201	0.21	0.33	0.19	0.54
Mixed culture	0.11	0.04	0.01	0.08

Treatment	Concentration (µg mL^-1)
Treatment	Acetic acid	Propionic acid	Butyric acid
Supernatant extracted from MRS broth	21.16 ± 0.13^* * Values represent mean ± standard deviation of three independent experiments. Different lowercase superscripts represent a significant difference using Duncan’s Test with a confidence level of 95%.	33.63 ± 0.10	4.89 ± 0.04
Supernatant extracted from MRS broth supplemented with honey	25.53 ± 0.11	39.73 ± 0.13	6.33 ± 0.11

Time (week)	Product 1: Rice grains		Product 2: Honey		Product 3: Rice grains & honey
Time (week)	Log cell number	Survival rate	Log cell number	Survival rate	Log cell number	Survival rate
0	9.08 ± 0.06^* * Values represent mean ± standard deviation of three independent experiments. Different lowercase superscripts represent a significant difference using Duncan’s test with a confidence level of 95%.	100.00	9.47 ± 0.26	100.00	8.82 ± 0.14	100.00
1	8.56 ± 0.10	94.27	9.79 ± 0.18	103.38	8.74 ± 0.08	99.09
2	7.91 ± 0.11	87.11	9.13 ± 0.12	96.41	8.59 ± 0.09	97.39
3	7.83 ± 0.04	86.23	8.69 ± 0.01	91.76	8.72 ± 0.18	98.87
4	7.19 ± 0.06	79.19	8.98 ± 0.08	94.83	8.29 ± 0.05	93.99
5	7.82 ± 0.01	86.12	6.50 ± 0.03	68.64	8.47 ± 0.02	96.03
6	7.27 ± 0.05	80.07	5.10 ± 0.06	53.85	8.28 ± 0.03	93.88
7	7.48 ± 0.01	82.38	3.89 ± 0.07	41.08	8.13 ± 0.03	92.18
8	7.63 ± 0.15	84.03	0.0 ± 0.0	0.00	7.96 ± 0.06	90.25

Test items	Unit per 100 g of product
Calories	364.26 kcal
Total fat	2.46 g
Saturated fat	0.72 g
Trans fat	Not detected
Cholesterol	Not detected
Sodium	13.71 mg
Carbohydrate	78.21 g
Dietary fiber	4.11 g
Sugar	12.04 g
Includes added sugars	1.00 g
Proteins	7.32 g
Calcium	21.50 mg
Iron	1.79 mg
Potassium	263.78 mg

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[1] *Corresponding author: saranpromsai@hotmail.com