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The potential of resistant starch as a prebiotic

Zaman SA, Sarbini SR

Crit Rev Biotechnol, 2016 Jun;36(3):578-84.
PMID: 25582732 DOI: 10.3109/07388551.2014.993590

Resistant starch is defined as the total amount of starch and the products of starch degradation that resists digestion in the small intestine. Starches that were able to resist the digestion will arrive at the colon where they will be fermented by the gut microbiota, producing a variety of products which include short chain fatty acids that can provide a range of physiological benefits. There are several factors that could affect the resistant starch content of a carbohydrate which includes the starch granule morphology, the amylose-amylopectin ratio and its association with other food component. One of the current interests on resistant starch is their potential to be used as a prebiotic, which is a non-digestible food ingredient that benefits the host by stimulating the growth or activity of one or a limited number of beneficial bacteria in the colon. A resistant starch must fulfill three criterions to be classified as a prebiotic; resistance to the upper gastrointestinal environment, fermentation by the intestinal microbiota and selective stimulation of the growth and/or activity of the beneficial bacteria. The market of prebiotic is expected to reach USD 198 million in 2014 led by the export of oligosaccharides. Realizing this, novel carbohydrates such as resistant starch from various starch sources can contribute to the advancement of the prebiotic industry.
Identification of acacia gum fermenting bacteria from pooled human feces using anaerobic enrichment culture

Rawi MH, Tan HY, Sarbini SR

Front Microbiol, 2023;14:1245042.
PMID: 37881253 DOI: 10.3389/fmicb.2023.1245042

Commercial acacia gum (AG) used in this study is a premium-grade free-flowing powder. It is a gummy exudate composed of arabinogalactan branched polysaccharide, a biopolymer of arabinose and galactose. Also known as food additive, acacia gum (E414), which is presently marketed as a functional dietary fiber to improve overall human gut health. The health effects may be related to the luminal pH regulation from the short-chain fatty acids (SCFA) production. Studies suggested that amylolytic and butyrogenic pathways are the major factors determining the SCFA outcome of AG in the lower gut. However, the primary bacteria involved in the fermentation have not been studied. This study aimed to investigate the putative primary degraders of acacia gum in the gut ecosystem. Isolation and identification of gum-fermenting bacteria were performed through enrichment culture fermentation. The experiment was conducted in an anaerobic chamber for 144 h in three stages. The study was conducted in triplicate using an anaerobic chamber system. This culture system allows specific responses to support only bacteria that are responsible for gum fermentation among the gut microbiota. Five bacterial strains were isolated and found to be gum-fermenting bacteria. Based on the 16s RNA sequence, the isolates matched to butyrate-producing Escherichia fergusonii, ATCC 35469.
In vitro fermentation of commercial α-gluco-oligosaccharide by faecal microbiota from lean and obese human subjects

Sarbini SR, Kolida S, Gibson GR, Rastall RA

Br J Nutr, 2013 Jun;109(11):1980-9.
PMID: 23116939 DOI: 10.1017/S0007114512004205

The fermentation selectivity of a commercial source of a-gluco-oligosaccharides (BioEcolians; Solabia) was investigated in vitro. Fermentation by faecal bacteria from four lean and four obese healthy adults was determined in anaerobic, pH-controlled faecal batch cultures. Inulin was used as a positive prebiotic control. Samples were obtained at 0, 10, 24 and 36 h for bacterial enumeration by fluorescent in situ hybridisation and SCFA analyses. Gas production during fermentation was investigated in non-pH-controlled batch cultures. a-Gluco-oligosaccharides significantly increased the Bifidobacterium sp. population compared with the control. Other bacterial groups enumerated were unaffected with the exception of an increase in the Bacteroides–Prevotella group and a decrease in Faecalibacterium prausnitzii on both a-gluco-oligosaccharides and inulin compared with baseline. An increase in acetate and propionate was seen on both substrates. The fermentation of a-gluco-oligosaccharides produced less total gas at a more gradual rate of production than inulin. Generally, substrates fermented with the obese microbiota produced similar results to the lean fermentation regarding bacteriology and metabolic activity. No significant difference at baseline (0 h) was detected between the lean and obese individuals in any of the faecal bacterial groups studied.
Fulltext Manipulation of Gut Microbiota Using Acacia Gum Polysaccharide

Rawi MH, Abdullah A, Ismail A, Sarbini SR

ACS Omega, 2021 Jul 20;6(28):17782-17797.
PMID: 34308014 DOI: 10.1021/acsomega.1c00302

Acacia gum (AG) is a branched-polysaccharide gummy exudate that consists of arabinose and galactose. The traditional practice in African-Middle Eastern countries uses this gum as medicine. Traditional use of AG is to treat stomach disease, which can be a potential functional food. In this research, commercially available AG from Acacia senegal and Acacia seyal was investigated as the prebiotic. The experiment employed a pH-controlled in vitro colon model inoculated with human fecal microbiota to mimic the human colon. Fermentation samples at 0, 6, 12, and 24 h were brought for short-chain fatty acid (SCFA) analysis using high-performance liquid chromatography and bacterial enumeration via fluorescent in situ hybridization. Results showed that AG significantly promotes Bifidobacteria proliferation similar to fructo-oligosaccharides (FOS) while inhibiting the Clostridium histolyticum group, commonly associated with gut dysbiosis. Acetate, propionate, and butyrate showed a similar trend to FOS (p > 0.05). The AG shows potential against gut dysbiosis, as it promotes gut-probiotics, through modulation of microbial population and SCFA production, especially butyrate.
Impact of phytic acid on nutrient bioaccessibility and antioxidant properties of dehusked rice

Lee HH, Loh SP, Bong CF, Sarbini SR, Yiu PH

J Food Sci Technol, 2015 Dec;52(12):7806-16.
PMID: 26604353 DOI: 10.1007/s13197-015-1918-9

Whole grains consumption promotes health benefits, but demonstrates controversial impacts from phytic acid in meeting requirements of good health. Therefore, this study was aimed to determine the nutrient bioaccessibility and antioxidant properties of rice cultivars named "Adan" or "Bario" and deduce the nutritional impact of phytic acid. Majority of the dehusked rice in the collection showed an acceptable level of in-vitro starch digestibility and in-vitro protein digestibility, but were poor in antioxidant properties and bioaccessibility of minerals (Ca, Fe and Zn). The drawbacks identified in the rice cultivars were due to relatively high phytic acid content (2420.6 ± 94.6 mg/100 g) and low phenolic content (152.39 ± 18.84 μg GAE/g). The relationship between phytic acid content and mineral bioaccessibility was strongest in calcium (r = 0.60), followed by iron (r = 0.40) and zinc (r = 0.27). Phytic acid content did not significantly correlate with in-vitro starch digestibility and in-vitro protein digestibility but showed a weak relationship with antioxidant properties. These suggest that phytic acid could significantly impair the mineral bioaccessibility of dehusked rice, and also act as an important antioxidant in non-pigmented rice. Bario rice cultivars offered dehusked rice with wide range of in-vitro digestibility of starch and protein, and also pigmented rice as a good source of antioxidants. However, there is a need to reduce phytic acid content in dehusked rice for improved mineral bioaccessibility among Bario rice cultivars.
Potential of novel dextran oligosaccharides as prebiotics for obesity management through in vitro experimentation

Sarbini SR, Kolida S, Deaville ER, Gibson GR, Rastall RA

Br J Nutr, 2014 Oct 28;112(8):1303-14.
PMID: 25196744 DOI: 10.1017/S0007114514002177

The energy-salvaging capacity of the gut microbiota from dietary ingredients has been proposed as a contributing factor for the development of obesity. This knowledge generated interest in the use of non-digestible dietary ingredients such as prebiotics to manipulate host energy homeostasis. In the present study, the in vitro response of obese human faecal microbiota to novel oligosaccharides was investigated. Dextrans of various molecular weights and degrees of branching were fermented with the faecal microbiota of healthy obese adults in pH-controlled batch cultures. Changes in bacterial populations were monitored using fluorescent in situ hybridisation and SCFA concentrations were analysed by HPLC. The rate of gas production and total volume of gas produced were also determined. In general, the novel dextrans and inulin increased the counts of bifidobacteria. Some of the dextrans were able to alter the composition of the obese human microbiota by increasing the counts of Bacteroides-Prevotella and decreasing those of Faecalibacterium prausnitzii and Ruminococcus bromii/R. flavefaciens. Considerable increases in SCFA concentrations were observed in response to all substrates. Gas production rates were similar during the fermentation of all dextrans, but significantly lower than those during the fermentation of inulin. Lower total gas production and shorter time to attain maximal gas production were observed during the fermentation of the linear 1 kDa dextran than during the fermentation of the other dextrans. The efficacy of bifidobacteria to ferment dextrans relied on the molecular weight and not on the degree of branching. In conclusion, there are no differences in the profiles between the obese and lean human faecal fermentations of dextrans.
Enzymatic hydrolysis: Sialylated mucin (SiaMuc) glycoprotein of edible swiftlet's nest (ESN) and its molecular weight distribution as bioactive ESN SiaMuc-glycopeptide hydrolysate

Hui Yan T, Lim SJ, Babji AS, Rawi MH, Sarbini SR

Int J Biol Macromol, 2021 Apr 01;175:422-431.
PMID: 33561458 DOI: 10.1016/j.ijbiomac.2021.02.007

Bioactive edible swiftlet's nest (ESN) sialylated-mucin (SiaMuc) hydrolysate is produced by alcalase hydrolysis. Enzymatic hydrolysis of ESN breakdown high-valued ESN SiaMuc-glycoprotein into bioactive SiaMuc-glycopeptide. This is a breakthrough for the issue of insolubility and low extraction rate in ESN, and even increases the bioavailability of ESN nutritional functionality and health benefits. Hydrolysis of ESN SiaMuc-glycoprotein was performed for 1 to 4 h and its effect on physicochemical properties, molecular weight (MW) distribution, SiaMuc-glycoprotein and glycopeptide integrity were determined. Other than improvement in solubility and bioavailability as SiaMuc-glycopeptide, results from SDS-PAGE revealed that MW of SiaMuc-glycoprotein decreased from 42.0-148.8 kDa to 17.7-142.7 kDa with increasing hydrolysis period. Further hydrolysis from maximized DH (90 min) showed an insignificant effect on the MW of ESN SiaMuc-glycopeptide and remained constant at 15.2 kDa. This highlights that enzymatic hydrolysis only influences macro SiaMuc-glycoprotein fractions (142.7, 115.3 and 102.7 kDa), while the majority of SiaMuc-glycopeptide fractions from 36.6-98.6 kDa remained intact. Conclusively, alcalase hydrolysis of ESN showed high recovery in the form of bioactive ESN SiaMuc-glycopeptide. Therefore, enzymatic biotechnology is an economic alternative applicable on ESN that broaden industrial utilization by reducing the MW without destroying the quality of bioactive SiaMuc-glycoprotein.
Fulltext Prebiotics metabolism by gut-isolated probiotics

Rawi MH, Zaman SA, Pa'ee KF, Leong SS, Sarbini SR

J Food Sci Technol, 2020 Aug;57(8):2786-2799.
PMID: 32624588 DOI: 10.1007/s13197-020-04244-5

There are numerous species of bacteria resides in the lumen of human colon. The word 'colon', resembles colony or the colonization of microbiota of which plays an important role in the fermentation of prebiotics. The standpoint of prebiotic nowadays is well reported for attenuating gut dysbiosis in many clinical studies tested on animals and human. However, because of the huge amount of gut microbiome, the attempt to connect the dots between bacterial population and the host are not plainly discernible. Thus, a need to analyse recent research on the pathways of prebiotic metabolism adopted by commonly studied probiotics i.e. Bifidobacteria and Lactobacillus. Several different substrate-dependent gene expressions are induced to break down oligosaccharide molecules shown by those probiotics. The hydrolysis can occur either by membrane bound (extracellular) or cytoplasmic (intracellular) enzyme of the enteric bacteria. Therefore, this review narrates several prebiotic metabolisms occur during gut fermentation, and metabolite production i.e. organic acids conversion.
Prebiotic evaluation of red seaweed (Kappaphycus alvarezii) using in vitro colon model

Bajury DM, Rawi MH, Sazali IH, Abdullah A, Sarbini SR

Int J Food Sci Nutr, 2017 Nov;68(7):821-828.
PMID: 28393631 DOI: 10.1080/09637486.2017.1309522

Red seaweed (Kappaphycus alvarezii) cultivated from Sabah (RSS) and Langkawi (RSL) were digested using in vitro mouth, gastric and duodenal model. The digested seaweed then fermented in a pH-controlled batch culture system inoculated with human faeces to mimic the distal colon. Bacterial enumeration were monitored using fluorescent in situ hybridisation, and the fermentation end products, the short chain fatty acids (SCFA), were analysed using HPLC. Both RSS and RSL showed significant increase of Bifidobacterium sp.; from log10 7.96 at 0 h to log10 8.72 at 24 h, and from log10 7.96 at 0 h to log10 8.60 at 24 h, respectively, and shows no significant difference when compared to the Bifidobacterium sp. count at 24 h of inulin fermentation. Both seaweeds also showed significant increase in total SCFA production, particularly acetate and propionate. Overall, this data suggested that K. alvarezii might have the potential as a prebiotic ingredient.
Umai Dish Preparation Practices and Food Safety Behaviors in Bintulu Locals on Borneo Island, Malaysia

Sukirman AN, Khalex HB, Mustafa S, Sarbini SR, Hassan S, S-Hussain SS, et al.

J Food Prot, 2020 Oct 01;83(10):1764-1774.
PMID: 32463874 DOI: 10.4315/JFP-19-543

ABSTRACT: Umai is a popular, traditional, native dish of the Melanau ethnic group in Sarawak. It is prepared using thin slices of raw marine fish marinated with calamansi juice and seasoned with other ingredients. The local people believe that the acidity of the citrus juice, along with the use of salt and spice, can slightly cook the fish and remove the fishy smell. The aim of this study was to investigate (i) the different umai handling and preparation practices and (ii) the personal experience of umai consumption among respondents. A purposive sample of 100 umai makers, divided into two equal groups, professionals and nonprofessionals, participated in the study. We found that Spanish mackerel and hairfin anchovy were ranked first and second in the list of species chosen for making umai, with the former mostly preferred by the professional group, as opposed to the latter, which was preferred by the nonprofessional group. Black pomfret was ranked third, where it is equally preferred by both groups. About 20% of respondents would freeze the raw fish chunks prior to preparing umai, as opposed to 26% who would sun dry their fish. Other techniques, such as salting and marinating (using calamansi juice), were also used during the preparation of umai. Most of the respondents indicated that they would consider the umai ready to eat soon after marinating (with all ingredients) the raw fish. One-third of both respondent groups indicated that they would chill the umai dish at 4°C for 30 min before serving. The respondents could not provide any rationale behind these food preparation practices. Overall, this study provides evidence of the different preparation methods for umai. These practices can thus be considered important targets for public health education campaigns seeking to improve food safety surrounding this food group.
Developments in understanding and applying prebiotics in research and practice-an ISAPP conference paper

Scott KP, Grimaldi R, Cunningham M, Sarbini SR, Wijeyesekera A, Tang MLK, et al.

J Appl Microbiol, 2020 Apr;128(4):934-949.
PMID: 31446668 DOI: 10.1111/jam.14424

AIMS: The concept of using specific dietary components to selectively modulate the gut microbiota to confer a health benefit, defined as prebiotics, originated in 1995. In 2018, a group of scientists met at the International Scientific Association for Probiotics and Prebiotics annual meeting in Singapore to discuss advances in the prebiotic field, focussing on issues affecting functionality, research methodology and geographical differences.
METHODS AND RESULTS: The discussion ranged from examining scientific literature supporting the efficacy of established prebiotics, to the prospects for establishing health benefits associated with novel compounds, isolated from different sources.
CONCLUSIONS: While many promising candidate prebiotics from across the globe have been highlighted in preliminary research, there are a limited number with both demonstrated mechanism of action and defined health benefits as required to meet the prebiotic definition. Prebiotics are part of a food industry with increasing market sales, yet there are great disparities in regulations in different countries. Identification and commercialization of new prebiotics with unique health benefits means that regulation must improve and remain up-to-date so as not to risk stifling research with potential health benefits for humans and other animals.
SIGNIFICANCE AND IMPACT OF STUDY: This summary of the workshop discussions indicates potential avenues for expanding the range of prebiotic substrates, delivery methods to enhance health benefits for the end consumer and guidance to better elucidate their activities in human studies.
Gastrointestinal parasitic infections of buffaloes (Bubalus bubalis) in Sarawak Borneo: Prevalence, risk factors, and farming practices

Harizt AM, Malahubban M, Syed-Hussain SS, Ramanoon SZ, Sadiq MB, Sarbini SR, et al.

Trop Biomed, 2021 Sep 01;38(3):318-326.
PMID: 34508339 DOI: 10.47665/tb.38.3.072

The objectives of this study were to investigate the prevalence and associated risk factors for gastrointestinal (GI) parasites in buffaloes from various areas of Sarawak, and to assess current management practices of GI parasites among farmers. Faecal samples were collected from 15 farms and 129 animals, as well as data on farm and animal-based characteristics. A total of 129 faecal samples were examined for GI parasites using a modified McMaster and sedimentation. Association between potential risk factors and the prevalence of GI parasites was investigated using Chi-square statistic. The prevalence of Paramphistomum sp., strongyles, and coccidia were 75.2% (95% CI±7.5), 52.7% (95% CI±8.6) and 48.1% (95% CI±8.6), respectively. Farms which had a grazing area less than 50 acres in size had significantly higher prevalence of strongyles (70.5%, χ2 = 8.34, P = 0.004) and paramphistomes (88.6%, χ2 = 6.46, P = 0.01) relative to farms with a larger grazing area (43.5% and 68.2%, respectively). Prevalence of strongyles was lower in farms that did not implement a cut- and-carry system (45.6%, χ2 = 4.17, P = 0.04) in comparison to those that did (64%). The prevalence of paramphistomes was higher on farms with more than 40 animals (80.6%, χ2 = 3.18, P = 0.05) relative to farms with fewer animals. The majority of farmers surveyed (67.9%) showed awareness of GI parasite infection and reported that they recognized the associated symptoms. Most farmers practised deworming, and ivermectin was the most commonly used anthelminthic (60.4%); only 1.9% of farmers used albendazole. Overall this study revealed a high prevalence of GI parasites in buffalo in Sarawak. Although farmers report they are aware of parasitic diseases, further education is still required. This could include how they can successfully implement on-farm changes to reduce the prevalence of GI parasites in their herds.

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