processes
Review
New Insights in Prebiotic Utilization: A Systematic Review
Martina Arapović 1 , Leona Puljić 1 , Nikolina Kajić 1 , Brankica Kartalović 2 , Kristina Habschied 3
and Krešimir Mastanjević 3, *
1
2
3
*
Citation: Arapović, M.; Puljić, L.;
Kajić, N.; Kartalović, B.; Habschied,
K.; Mastanjević, K. New Insights in
Prebiotic Utilization: A Systematic
Review. Processes 2024, 12, 867.
Faculty of Agriculture and Food Technology, University of Mostar, Biskupa Čule bb,
88000 Mostar, Bosnia and Herzegovina; martina.arapovic@aptf.sum.ba (M.A.);
leona.puljic@aptf.sum.ba (L.P.); nikolina.kajic@aptf.sum.ba (N.K.)
Institut Biosens, Zorana Ðind̄ića 1, 21000 Novi Sad, Serbia; brankica.kartalovic@biosense.rs
Faculty of Food Technology Osijek, J.J. Strossmayera University of Osijek, F. Kuhača 18, 31000 Osijek, Croatia;
khabschi@ptfos.hr
Correspondence: kmastanj@gmail.com
Abstract: The hectic pace of modern life often leads to quick solutions, both in lifestyle and the choice
of food we consume. The importance of the gut microbiome and its balance is being increasingly
researched, with the prebiotic concept itself becoming a topic of scientific investigation. The aim of
this paper is to analyze scientific studies on the understanding of prebiotics conducted between 2019
and 2024 in order to see what new knowledge, new sources, new ways of use, and newly established
effects on certain disease states have been discovered during this period. The question that the
authors are trying to answer is how specific prebiotics affect the growth and activity of selected
probiotic strains in the human gut (have impact on gut microbiome) and what the implications of
these interactions are. Four databases were searched: Pubmed/MEDLINE, Springerlink, Google
Scholar, and Scopus. The keywords used were prebiotics, functional food, probiotics, gut microbiome,
and trends. A systematic review of 30 scientific studies on the topic of prebiotics revealed significant
advances in understanding and application. Research particularly indicates how prebiotics stimulate
the growth of beneficial probiotic strains, such as Lacticaseibacillus rhamnosus, Lactiplantibacillus
plantarum, and Bifidobacterium. In addition, innovative approaches in food production, including
pasta rich in prebiotic fibers, chocolate with inulin and stevia, and the use of fruit by-products, show
promising results in creating “healthier” food options. Although the papers had differing objectives
and research methodologies, certain similarities were found. All papers emphasized the importance
of using prebiotics, although it depended on the type they come from and their impact on the gut
microbiome, i.e., the stimulation of probiotic action within the gut microbiome, which consequently
has benefits on health. This review serves as a springboard for further research in this exciting field,
with the ultimate goal of harnessing the power of prebiotics to improve health outcomes.
https://doi.org/10.3390/pr12050867
Keywords: prebiotic; probiotics; gut microbiome; functional food
Academic Editor: Dariusz Dziki
Received: 5 April 2024
Revised: 18 April 2024
Accepted: 23 April 2024
Published: 25 April 2024
Copyright: © 2024 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
1. Introduction
“Let food be thy medicine, and medicine be thy food” (Hippocrates)
In the last decade, prebiotics have attracted significant attention in scientific research
due to their role in stimulating the intestinal microflora in the degradation of nutrients and
improving the overall health status of humans. Their degradation products are short-chain
fatty acids (SCFA) that are released into the bloodstream, consequently affecting not only
the gastrointestinal tract but also other distant organs [1]. Through the effort and work
of scientists, prebiotics today, after numerous studies thanks to the Consensus Panel of
the International Scientific Association for Probiotics and Prebiotics (ISAPP), are defined
as follows: “Substrates that are selectively utilized by host microorganisms conferring a
health benefit” [2]. The concept of prebiotics was first introduced by Glenn Gibson and
Marcel Roberfroid in 1995 [3]. Only a few compounds from the group of carbohydrates,
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https://www.mdpi.com/journal/processes
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such as short-chain and long-chain β-fructans (FOS and inulin) and lactulose or galactooligosaccharides (GOS), can be classified as prebiotics. By modulating the gut microbiota,
prebiotics stimulate the growth of “good bacteria” probiotics, such as Lactobacillus and
Bifidobacterium, while reducing the growth of those “bad bacteria” or pathogens [4]. By
emphasizing research and raising people’s awareness of proper and balanced nutrition, the
problem of the onset of related diseases is sought to be suppressed. Thus, the aim of this
paper is to analyze the latest findings on the prebiotic potential and answer the question
of how specific prebiotics affect the growth and activity of selected strains of probiotics in
digestion (in vitro and in vivo) and what the implications of these interactions are. This
will be investigated by reviewing the literature and studying papers in databases relevant
to the given subject of research. Also, the aim is to identify key knowledge gaps in the field
of prebiotics and topics that are currently of interest to scientists, such as optimal dosages,
their interactions with probiotics that potentially result in the greatest health benefits, and
similar topics.
2. Existing Knowledge: Types of Prebiotics, Their Sources, and Impact
In the next section, we will cover the most frequently mentioned types of prebiotics and
their sources that have been mentioned in further research. In addition to the mentioned
ones, other known groups of potential prebiotics are oligosaccharides derived from starch
and glucose, where resistant starch belongs, which is an indigestible component of human
intestines, and other oligosaccharides, such as pectin (pectin oligosaccharide—POS), for
example, flavanols from cocoa, which belong to oligosaccharides without carbohydrates
and have shown their potential by acting on the stimulation of lactic acid bacteria.
2.1. Fructans
Fructans are a type of prebiotic composed of inulin and fructooligosaccharides (FOS)
or oligofructose. Their structure is a linear chain of fructose with a β (2→1) bond. They
usually consist of glucose units with a β (2→1) bond at the end [5]. Initial research pointed
to the selective stimulation of fructans on lactic acid bacteria, but in recent years, more
and more emphasis is being placed on chain length as an important factor influencing the
determination of the type of bacteria that can ferment [6].
2.1.1. Inulin
Inulin-type fructans are polymers made from fructose linked to terminal α-linked
glucose via a β (2→1) bond. Chicory is considered the richest source of inulin. Several
studies have shown that inulin-type fructans can enhance the growth of bifidobacteria and
Anaerostipes [7]. Inulin is most commonly used as a fat substitute [8]; it has been proven to
enhance mineral absorption in the body [9,10], alleviate constipation [11], and promote the
growth and development of Lactobacillus and Bifidobacterium strains within the colon [12],
which is why it is important to this study.
2.1.2. FOS
Fructooligosaccharides (FOS) are a researched type of prebiotic that can be found in
bananas, wheat, onions, and garlic. The main component of the FOS structure in β (2→1)
bonds is fructose. FOS derivatives are beneficial for colon health, as they promote the
selective growth of bifidobacteria and lactobacilli and potentially inhibit microorganisms in
intestinal cells, enhance the maintenance of body bacterial balance, alleviate constipation,
and maintain healthy gut flora [13].
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2.1.3. GOS
Galactooligosaccharides (GOSs), a product of lactose degradation, are classified into
two subgroups: GOS with an excess of galactose at C3, C4, or C6 and GOS produced from
lactose by enzymatic transglycosylation. The end product of this reaction is mainly a mixture of tri- to pentasaccharides with galactose in β (1→6), β (1→3), and β (1→4) bonds. This
type of GOS is also called trans-galacto-oligosaccharides or TOS [14]. GOS is considered
one of the most researched types of prebiotics and is safe for consumption. Galactooligosaccharides can greatly stimulate the growth of bifidobacteria and lactobacilli [15]. Its most
common use currently is in infant milk formula, where it has proven to be a help in gut
modulation by promoting the growth of Bifidobacterium strains in infants who cannot be
fed breast milk [16].
2.2. Newly Recognized Unclassified Prebiotics
With the advancement of technology, prebiotic preparation methods have been optimized. In addition, various new types of prebiotics have been developed (mainly including
polysaccharides, polyphenols, and polypeptide polymers) [17]. Emerging prebiotics are
mainly found in algae, fruit juices, and fruit such as chokeberry [18]. Although knowledge
about them is not yet as established as about prebiotic types FOS and GOS, their potential
deserves in-depth study and has a promising future. The mentioned research is located in
Section 4.
2.3. Importance of Prebiotic Interaction on Probiotic Growth and Stimulation Mechanism
The human digestive system, often described as the “second brain,” plays a crucial
role in maintaining health. The gut microbiome consists of approximately 3.9 × 1013
bacteria in the large intestine, composed of probiotics, neutral bacteria, and pathogenic
bacteria [19], which compete with each other to establish a balance in order to prevent the
development of diseases. Scientists are constantly searching for new sources of prebiotics
and probiotics and their synergistic action, as diseases related to gut balance disorders,
such as obesity, type 2 diabetes, irritable bowel syndrome, various mental disorders, and
colon cancer, are in constant worrying increase. The notion of probiotic food often brings
to mind mild kefir and fermented dairy food [20]. The gut microbiome is in constant
interaction with the human body, helping the body digest and absorb nutrients from food,
metabolize toxic waste produced in the intestines, and produce functional substances
necessary for life. This primarily refers to amino acids, vitamins, short-chain fatty acids
(SCFA), and other substances [21]. Since probiotics are live microorganisms that enhance gut
function with sufficient intake, their growth and reproduction require the intake of mainly
polysaccharides that the body cannot digest—prebiotics. After the intake of a prebiotic
compound, it passes through the small intestine and reaches the large intestine undegraded.
Here, probiotic microorganisms break them down primarily into SCFAs, which later act on
the organism, using fermentation. Firmicutes and Bacteroidetes are physiologically dominant
species in the fecal microbiota of the adult large intestine, while Actinobacteria predominates
in infants [22]. The ratio of Firmicutes to Bacteroides was often considered a trigger for
obesity, but newer meta-analyses refute this theory. Unlike studies done on mice, human
studies have not confirmed these results. Disagreements between these studies highlight
the lack of reliable microbial taxonomic indicators for obesity [23]. Further research is
needed to establish this fact. Furthermore, research indicates that the composition of the
gut microbiome may be related to personality traits [24]. Among probiotic strains, the most
commonly described are strains of Lactobacillus (belongs to the phylum Firmicutes) and
Bifidobacterium (belongs to the phylum Actinobacteria) [22]. In addition to them, this group
also includes Enterococcus, Streptococcus, and Escherichia. The fungus Saccharomyces boulardii
is not naturally found in the gut microbiome, but its supplementation is of particular
interest to people who need to replace gut dysbiosis caused by the use of antibiotics [25].
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3. Materials and Methods
Due to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses),
guidelines were followed in this review. Search was performed from inception until January 2024 in Pubmed/Medline, Springerlink, Google Scholar, and Scopus databases for
randomized clinical trials that assessed the effects of prebiotic type on human gut in vitro
and in vivo among adults and children and that were published in English language.
3.1. Inclusion Criteria
The research encompasses insights into the effects of prebiotics based on clinical trials,
specifically randomized controlled trials (RCTs). The target population was adults and
children with specific health conditions, comparing the composition of the gut microbiota
and its impact on health before and after the study. The studies include comparisons of the
type of prebiotic sources used and their impact on health. The consequences of prebiotic
action, including improvement, stimulation of probiotic growth, and the duration of the
study, are taken into account. The time span of the studies considered was within the last
5 years (2019–2024).
3.2. Exclusion Criteria
All studies not conducted on humans were excluded (animals, review papers, study
protocols, or observational studies). Case studies were also not included. Clinical trials
that did not have a precisely defined type of prebiotic used and an examination of their
subsequent action either on the gut microbiome or on the stimulation of probiotic strain
growth in the gut microbiome were not considered. Papers that did not belong to this study
by title, i.e., included a type of research that is not the subject of inclusion, were excluded.
The duration of the studies was not limited.
3.3. Research Process
Four databases were utilized for the search: PubMed/MEDLINE, Scopus, Springerling,
and Google Scholar. The keywords used for Google Scholar were “prebiotics”, “functional
food”, “probiotics”, “gut microbiome”, “trends”. During the Scopus database search, the
“Food Science” filter was applied. After the search, a database of 6120 scientific papers was
obtained. Prior to the search, papers that overlapped (n = 3355), were not originally written
in English (n = 26), and did not belong to the range of research subjects by year (n = 1506)
were excluded. Through the processing of the obtained papers (n = 1233), studies conducted
on animals (n = 236), non-randomized controlled trials (n = 76), those not relevant to the
title of this systematic review (n = 99), and those without access enabled (n = 592) were
excluded. Inclusive, this study covered 53 literature sources, of which 30 scientific papers
were taken for analysis.
4. Results and Discussion
The investigated articles are listed in Tables 1 and 2, and the PRISMA flowchart is
presented in Figure 1. Based on the stated inclusion criteria for the research, the authors
have arrived at the following observations.
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Figure 1. PRISMA flow diagram used for research.
4.1. Impact of Prebiotics from New Sources on the Growth of Specific Probiotics
The act of choosing and utilizing food does not need to be intricate. In their research, [26] tested the ways how different concentrations of apple, banana, and mango peel
powder affect the growth of probiotics. In this way, they tried to utilize the by-products of
fruit processing. They concluded that a small amount of purified fruit peel powder at a
concentration of 2% shows prebiotic action by stimulating the growth of Lactocaseibacillus
rhamnosus (LGG), L. casei, and B. animalis subslactis (BB-12), proving that the by-product
can be commercially utilized. Furthermore, researching purified banana peel powder, [27]
enriched cookies with fibers while controlling the growth rate parameter of the probiotic
strain Lactobacillus spp. They found a significantly higher number of colonies compared to
the controlled groups of inulin, glucose, and the placebo. Further research is anticipated
on the organoleptic assessment of this type of cookie. Aiming to exploit the potential of
radish leaf by-products, ref. [28] investigated its potentials as prebiotics. Specifically, plant
extracts show promise as potential therapeutics for the prevention of metabolic syndrome
and obesity. The composition of immature radish extract, i.e., the prebiotic activity of RGP
(radish green polysaccharide), was analyzed. Prebiotic action was established, resulting
in changes in the concentration of SCFA and the pH of the gut microbiome. Probiotic
strains successfully utilize RGP, proving its potential as a prebiotic. In addition, research
conducted with oral consumption of the probiotic strain Lactiplantibacilus plantarum P-8,
in combination with RGP, resulted in an increase in the probiotic strain and the inhibition
of fat accumulation in adipocytes. The final results led to a reduction in fat accumulation
in the body, which could effectively reduce obesity. This study proves that RGP, as a
polysaccharide, promises as a natural prebiotic ingredient in the food industry. With the
aim of proving polyphenols as prebiotics, in a 12-week study conducted on a group of
healthy young men [18], the subjects consumed aronia extract or whole aronia fruit powder,
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which had been previously purified. The research findings indicate a positive impact on
the potential consumption of polyphenols from aronia for maintaining the cardiovascular
health of the young men studied. In an attempt to demonstrate the prebiotic effect of five
types of legume honey and the probiotic effect on L. rhamnosus and L. paracasei, ref. [29]
used astragalus, carob, alfalfa, sainfoin, and indigo honey in their research. They concluded
that the polyphenols present in these types of honey show a prebiotic potential on L. rhamnosus and L. paracasei strains with antagonistic action towards the five most dangerous
pathogens depending on the type of honey. The authors look forward to further research
on the postbiotic action of legume honey. Fucoidan (purity ≥ 95%), isolated from brown
algae, attracted the attention of [30], so they investigated its prebiotic potential. In their
research, finding an optimal concentration of 8 mg/mL, they proved the regulation of the
antibacterial ability of L. rhamnosus and noticed a great concentration dependence on the
action. They observed the growth inhibition zone. Unlike them, ref. [31] concluded that the
water extract of P. ostreatus, obtained via extraction from white oyster mushroom, stands
out due to its β-glucan content. Namely, water-soluble β-glucan from P. ostreatus proved a
prebiotic potential by stimulating the growth of L. plantarum, creating a symbiotic. Ref. [32]
investigated the prebiotic properties of a sample of purified rice bran extract (RB) on the
stimulation of the growth and the inhibition of gastrointestinal probiotic pathogens. As
a parallel control, they used inulin and distilled water. They followed the growth of the
genera L. casei and L. plantarum. The results of this study indicate that the rice bran extract
obtained by xylanase (RB1) contains a higher concentration of oligosaccharides that serve
as a prebiotic compound alongside those obtained with water (RB2). This study predicts
another possibility of using by-products from the food industry with the aim of creating
functional food. Trying to mitigate the negative effect of sugar in chocolate but not to
eliminate it from the diet, ref. [33] conducted research on chocolate with a low content of
fat, sugar, phytochemicals, and prebiotics, making chocolate from cocoa, inulin, and stevia.
The more acceptable version of chocolate proved to give the best sensory experience to
consumers based on the hedonic scale. Furthermore, the polyphenols, flavonoids, and
alkaloids coming from cocoa powder were noted as good sources of phytochemicals and
contributed to the benefits. The addition of inulin together with phytonutrients gave the
product prebiotic properties and stimulated the growth of L. rhamnosus. In addition, inulin
was used to limit the caloric value acting as a good substitute for fat. The glycemic index
was limited since stevia was used as a sweetener. This product promises to be a healthier,
prebiotically richer version of chocolate, and future research is expected.
Table 1. Impact of prebiotics from new sources on the growth of specific probiotics.
Author
Prebiotic Source
Hafza Fasiha Zahid
et al. (2021)
[26]
Chee Yee Tan et al.
(2024)
[27]
Florinda Fratianni et al.
(2023)
[29]
Concentrations
Probiotic Type
Impact
Apple, banana, and
mango peel powder
0%, 2%, and 4%
Lacticaseibacillus
rhamnosus,
L.casei, Bifidobacterium
lactis
↑ lactic acid bacteria
(LAB).
Banana peel powder,
Plantain Peel Powder,
inulin
10%, 20%, and 30%
Lactobacillus spp.
↑ Lactobacillus spp.
-
Lacticaseibacillus casei,
Lactobacillus gasseri,
Lacticaseibacillus
paracasei subsp.
Paracasei,
Lactiplantibacillus
plantarum,
Lacticaseibacillus
rhamnosus
Improve prebiotic
properties; have
inhibitory biofilm effect
Legumes honey’s
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Table 1. Cont.
Author
Prebiotic Source
Concentrations
Probiotic Type
Impact
Yanli Zhu et al. (2021)
[30]
Fucoidan as prebiotic,
isolated from Undaria
pinnatifida
0, 0.8, 8,
80 mg/mL,
L. rhamnosus
Inhibiting bacterial
infections
Anindita Deb Pal et al.
(2023)
[33]
Prebiotic Enriched
(inulin) Chocolates
2% chocolate extract
and 1% sub-cultured
broth
Lacticaseibacillus
rhamnosus
May promote
well-being consequent
to alternative of
conventional chocolates
↑ Lacticaseibacillus,
Lactobacillus, and
Bifidobacterium
↑ SCFA
Yu Ra Lee et al. (2023)
[28]
Radish
(Raphanus sativus L.)
4.9%
B. bifidum, B. Longum,
Lacticaseibacillus
paracasei,
Lactiplantibacillus
plantarum, Escherichia
coli, Lactobacillus lactis
Ryan Haryo Setyawan
et al. (2023)
[31]
White Oyster
Mushroom
(P. ostreatus) Extract
2% w/v and
0.25% w/v
Lactiplantibacillus
plantarum
and pathogen
Escherichia coli
↑ L. Plantarum Dad-13
Istas et al. (2019)
[18]
Aronia berry whole
fruit (WF),
extract (EX),
500 mg/capsule/day
↑ in Anaerostipes
↑ in Bacteroides
Significantly improved
endothelial function
1% v/v
L. casei, L. Plantarum,
and pathogen B. cereus,
E. coli
↑ L. casei, L. plantarum,;
high potential for
inhibiting the growth
of pathogenic B. cereus
and E. coli.
Thornthan
Sawangwani et al.
(2024)
[32]
Rice bran extract
↑—increasing value.
4.2. Correlation of the Use of Prebiotics from New Sources and Human Health Status
4.2.1. The Impact of Consuming Fructans as Prebiotics
In an attempt to prevent the diseased states of the “modern age”, there is increasing
research in the correlation between gut microbiota, its composition, and its impact on
human health. Hectic lifestyles make it difficult to properly choose and access “healthy”
foods, so researchers are trying to discover new, modify existing, and make “healthier”
versions of popular foods. In the search for healthy food, ref. [34] researched the impact
of dragon fruit oligosaccharides on gut microbiota. The research proved prebiotic activity
and the modulation of the gut microbiome. A small dose of just 4 g/day had an impact on
IgA, while a dose of 8 g/day successfully promotes the growth of the probiotic strains Bifidobacterium spp. and Faecalibacterium while inhibiting the growth of Escherichia coli strains.
Conducting a double-blind RCT study, where FOS was used as a placebo and a parallel
encapsulated combination of FOS with Lactiplantibacillus plantarum and Bifidobacterium breve,
ref. [35] interestingly, concluded that despite increased fat intake, markers associated with
obesity improved in the group where the combined version of prebiotics and probiotics
was consumed. This evidence provides important information on the possible use of
prebiotics in the prevention of obesity, but further research is still required. Investigating
the correlation between the intakes of the antipsychotic olanzapine in individuals suffering
from schizophrenia and weight gain, ref. [36] attempted to prevent this side effect. By
setting up two studies, they examined the effect of olanzapine on the consumption of the
probiotic itself and olanzapine and probiotic with a combination of 10 g of bitter melon
(Momordica charantia) and oligosaccharides. It was concluded that this combination effectively reduces the weight gain caused by the use of olanzapine while retaining the desired
psychopathological effects. Probiotics alone were not sufficient to prevent the side effect
of weight gain caused by the use of this drug. Also, it was discovered that the addition
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of a combination of probiotics and prebiotic fibers of bitter melons have a significant benefit for insulin resistance. Striving to make a modified and “healthier” version of pasta,
ref. [37] proved to be able to lower the glycemic index by adding barley β-glucans and
probiotic spores BC30. This study shows for the first time that basic food such as pasta
can be modified into a symbiotic food that can have beneficial effects in people at risk of
developing diseases due to an unhealthy lifestyle. This opens up possibilities for designing
new “healthier” alternatives to basic foods. Unlike [37], in the research of [38], the impact
of barley β-glucans on lipid metabolism in patients at high risk of developing metabolic
syndrome was monitored. B-glucans influenced the formation of short-chain fatty acids,
a change in the composition of the gut microbiota with an increase in the diversity of
microorganisms in accordance with previous observations conducted on animal models,
primarily Bifidibacteriae. The need for a “personalized” approach to nutrition has also been
concluded. Comparing the prebiotic properties of β-glucans from oats and caffeine-free
extracts from green coffee on obesity and its comorbidities, ref. [39], conducting research
on individuals with excessive body weight, concluded that regular consumption of the
four nutraceuticals based on the combination of oat beta glucans and prebiotics from green
coffee may reduce cardiovascular risk and help in preventing type 2 diabetes and obesity.
By supplementing oligofructose-enriched p-inulin, ref. [40] researched its impact on the
metabolic response of the gut in people with chronic kidney disease (CKD). The results
suggest a reduction in carbohydrate metabolism in the subjects, which led to a change in the
gut microbiome and consequently resulted in an increase in Bifidobacteria and Anaerostipes.
Trying to create a non-pharmacological drug for people suffering from respiratory diseases,
ref. [41] researched the impact of adding inulin to inflammatory indicators in obese people
with asthma. They proved the potential of using inulin alone, unlike that with the addition
of probiotics, which interestingly had fewer benefits than inulin alone. The authors look
forward to further research on this non-pharmacological drug. Pointing to the persistence
of the link between strength and muscle function in older people and the use of prebiotics,
ref. [42] gave their subjects a prebiotic supplement (inulin and fructo-oligosaccharide).
Healthy older twin pairs were used in the examination. They concluded the prebiotic effect
on improving cognition in the elderly, but they did not have an impact on strength and
muscle function compared to the placebo, as expected. With their research on the impacts
of adding the prebiotic fiber inulin (fructooligosaccharide), Vitafiber (isomaltooligosaccharide), and Fibremax (mixture of different fiber) to the diet of healthy women, [43] proved
that the greatest impact was from the addition of inulin on the intestinal microflora and the
increase in the strain Bifidobacterium. Further research is needed to individually determine
the selection of the most suitable prebiotic fibers for maximum utilization in the formation
of short-chain fatty acids.
4.2.2. The Impact of Consuming GOS as Prebiotics
An extremely high-quality study proved the prebiotic potential of their objects of
research, GOS, MOS, GMOS, and GMPS, by [44] showing prebiotic effects for C. butyricum
and bifidobacteria. In conclusion, different prebiotic effects of galactosyl and mannosyl
carbohydrates have been proven, as well as the production of SCFA and potential symbiotic
effects, and a connection to their physical–chemical characteristics has been established.
An in vivo study, compared to commercial GOS and MOS, showed that hydrolyzed galactomannans with specific degrees of polymerization have special advantages in prebiotic
effects and display potential as new prebiotic products. The first study that followed the
correlation between the intake of prebiotic galactooligosaccharides (GOS) and the gut
microbiome of children with autism was conducted by [45]. A significant increase in the
probiotic strain Bifidobacterium was determined after consumption when compared to the
placebo group, but there were no changes in the spectrum of children’s behavior. Although
there was no significant difference in the results between the groups, the absence of side
effects and the discovered medium effects of the study were encouraging and require
further research. Ref. [46], in their double-blind, placebo-controlled, 4-week study of galac-
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tooligosaccharide (GOS) supplement interventions, sought to characterize the impact of
GOS on indices of emotional well-being and gut microbial composition in a sample of participants at the end of adolescence and in early adulthood. It was found that GOS increases
the abundance of Bifidobacterium in 4 weeks in tandem with a reduced manifestation of
anxiety and indications of modifying attention bias in a sample of adolescent women. The
presented data promise that GOS could be effective in influencing the reduction of anxiety.
4.2.3. Newly Recognized Unclassified Prebiotics and Their Impact on Health
In an attempt to exploit pectooligosaccharides (POS) from lemon peels as prebiotics,
ref. [47], compared with the standard prebiotic fructooligosaccharides (FOS) in older people
by researching in vitro fermentation, proved a greater potential of POS in the production of
short-chain fatty acids (SCFA) and a smaller proportion of the formation of branched-chain
fatty acids (BCFA) from the FOS group. The potential prebiotic properties of black and
white pepper were investigated by [48] compared with inulin as a positive control, and it
was concluded that black and white pepper shows significantly higher prebiotic properties
by promoting the growth of bifidobacteria compared to inulin. The authors attribute this
to the component of arabinogalactan present in Piper nigrum. In line with the research on
prebiotic potentials of spices, ref. [49] concluded that the substrates present in Curcuma
longa (turmeric), Zingiber officinale (ginger), Piper longum (pipli or long pepper), and Piper
nigrum (black pepper) may drive beneficial alterations in gut communities, thereby altering
their collective metabolism and contributing to the salubrious effects on digestive efficiency
and health. Researching the potential of black rice [50] as a prebiotic, the authors concluded
that the black rice extract contains 71.9 mg of anthocyanins per 100 g of dry weight of the
extract, so it could be used as a prebiotic ingredient in the gut microbiota in obese people.
The research was prompted by the fact that the consumption of 70.7 mg of anthocyanins
daily during 8 weeks of consumption favorably affects the composition of gut bacteria.
It has been established that there is a connection between high levels of anthocyanins to
a reduction in inflammatory markers and changes in gut microbiota and the prevention
of obesity in a way that produces strains of Lachnospiraceae and Ruminococaceae. These
strains in turn produce butyrate, which plays a crucial role in increasing the protection
of the intestinal barrier, thereby inhibiting the entry of anti-inflammatory molecules into
the bloodstream and preventing the onset of metabolic endotoxemia. By researching
the potential of arabinoxylan-oligosaccharides (AXOS) from wheat bran extract, ref. [51]
found that the intake of wheat bran extract increases fecal Bifidobacterium and softens the
consistency of the stool without major effects on energy metabolism in healthy people with
slow GI transit. By proving the potential of resistant starch as a prebiotic, ref. [52] discovered
the benefits of resistant corn starch that are associated with changes in the composition
of the gut microbiota. These changes in the gut microbiota affect the bile acid profile,
reduce inflammation by restoring the intestinal barrier, and inhibit lipid absorption. As a
potential prebiotic polyphenol from Montgomery cherry, ref. [53] demonstrated an increase in
Lactobacillus strains in 22/28 participants, but the examined middle-aged human population
did not have significant changes in insulin responses as expected. However, there is still a
lack of information in the literature about optimal bioactive doses for modifying the gut
microbiome and managing the glucose response.
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Table 2. Correlation of the use of prebiotics from new sources and human health status.
Author
A Source of Prebiotics
Participants
Type of Research
Dose
Period
Conclusions
Nattha Pansai et al.
(2023)
[34]
Dragon fruit
oligosaccharides (DFO)
107
Randomized
Double-Blind, Placebo
Controlled Study
4 g and 8 g
4 weeks
↑ Bifidobacterium spp.;
decreased harmful bacteria,
especially, Escherichia coli.
DFO improved the immune
system
(serum Ig A) at even a low
dose (4 g/day)
Eun-Ji Song et al. (2020)
[35]
Fructo-oligosaccharide as
prebiotic; B. breve CBT BR3,
L. plantarum CBT LP3 as
probiotic
50
Double-blind,
placebo-controlled,
randomized clinical trial
2 tablets
12 weeks
Significantly improved
obesity-related markers in
obese people
15
Nonrandomized,
open-label, 3-phase pilot
trial, with repeated
measures within each
phase
8 g/twice a day
28 weeks
↑ Bifidobacterium and
Anaerostipes; increase in
abundance of microbial
metabolites derived from
carbohydrate metabolism.
Two sequential,
randomized clinical
trials
Study 1: olanzapine
(15–20 mg/day) plus
probiotics (840 mg twice
daily) vs. olanzapine
monotherapy
Study 2: olanzapine
(15–20 mg/day) plus
probiotics (840 mg twice
daily) and dietary fibre
(30 g twice daily) vs.
olanzapine monotherapy
12 weeks
Probiotics plus dietary fibre
significantly attenuated
olanzapine-induced weight
gain; Psychopathological
symptoms improved in all
groups
Michael B. Sohn et al.
(2024)
[40]
Oligofructose-enriched
inulin (p-inulin)
Huang et al. (2022)
[36]
Probiotic: Bifico containing
live Bifidobacterium,
Lactobacillus and Enterococcus
Prebiotic: bitter melon
(Momordica charantia) and
oligosaccharides
(fructooligosaccharides and
oligoisomaltoses), kudzu
starch, insulin and resistant
dextrin
Study 1, 90
Study 2, 60
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Table 2. Cont.
Author
A Source of Prebiotics
Participants
Type of Research
Dose
Period
Conclusions
Beatriz Míguez et al.
(2020)
[47]
Pectooligosaccharides
obtained from lemon peel
(POS)
6
In vitro experiments
6.5 g/day of FOS, POS or
SIEM
3 days
↑ SCFA and the lowest
BCFA; results support the
potential of pectin-derived
oligosaccharides as prebiotic
candidates targeting gut
health in the elderly
Mattea Müller et al.
(2020)
[51]
Wheat bran extract
Arabinoxylan
Oligosaccharide (AXOS)
48
Randomized,
Placebo-Controlled,
Double-blind study
15 g/day AXOS
12 weeks
↑ Bifidobacterium and softens
stool
Mary Ni Lochlainn et al.
(2024)
[42]
Prebiotic supplement (inulin
and fructo-oligosaccharides)
36 twin pairs
(72 individuals)
Placebo Controlled
Double-Blinded
Randomised Controlled
Trial
7.5 g of prebiotic: inulin
(3.375 mg) and FOS
(3.488 mg)
12 weeks
May improve cognition in
ageing population
Jian Tan et al. (2023)
[43]
Inulin
(fructo-oligosaccharide),
Vitafiber
(isomalto-oligosaccharide),
and Fibremax (mixture of
different fiber)
28
Cross-over intervention
Study
Three times daily, a total
of 15 g of Vitafiber, 34 g
of Fibrema, or 15 g of
inulin
3 weeks
Inulin supplementation had
highest impact on SCFA
production.
17
Randomised,
double-blinded, placebo
controlled 3-way
cross-over trial
3 × day
(inulin 12 g/day),
soluble fibre + probiotic
(inulin 12 g/day+
multi-strain
probiotic N25 billion
CFU) and placebo.
1 week
Improvements in airway
inflammation, asthma
control and gut microbiome
composition following
inulin supplementation
41
Single-Blind, Parallel,
Randomized,
Placebo-controlled
Dietary Intervention
Study
12 weeks
Affected glycemia- and
lipid-related markers;
resistin in a subgroup of
healthy obese or
hyperglycemic volunteers
Rebecca McLoughlin
et al. (2019)
[41]
Soluble fibre
supplementation (inulin)
with a probiotic
Donato Angelino et al.
(2019)
[37]
Whole-grain pasta
containing barley β-glucans
and Bacillus coagulans BC30,
6086
1 serving/d
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Table 2. Cont.
Author
A Source of Prebiotics
Ana Velikonja et al.
(2019)
[38]
Barley beta glucans
Raquel Mateos et al.
(2022)
[39]
Oat Beta-Glucan;
hydroxycinnamates,
caffeine-free extract from
green coffee
Siti Maisarah Nashri
et al. (2023)
[48]
Piper nigrum L.
Christine T. Peterson at
al. (2019)
[49]
Curcuma longa (turmeric),
Zingiber officinale (ginger),
Piper longum (pipli or long
pepper), and Piper nigrum
(blackpepper)
Lear et al. (2019)
[53]
Montmorency cherry (MC)
drink
Rachmat Faisal Syamsu
et al. (2023)
[50]
Black Rice Extract (Oryza
Sativa L.)
Participants
Type of Research
Dose
Double-blind,
placebo-controlled,
randomised clinical trial
6 g/day
60
Randomized,
dose–response, parallel,
blind study
3 g d−1 or 5 g d−1 doses
of 35% or 70% BG and a
fixed amount of GCBE
providing
600 mg d−1 of phenols.
3
In vitro colon
fermentation compared
with inulin effect
43
-
In vitro anaerobic fecal
cultivation
1%
28
RCT, DB, placebo
30 mL 2×/day
AC—296 mg/day
PP—1040 mg/day
33
Randomized control
trial, placebo research
with a pre-post test
approach
Black Rice Extract
5.6 g/day
12
Period
Conclusions
4 weeks
Affect lipid
509 metabolism in patients
with high risk for MS
development; effect on
glucose
510 metabolism was not
confirmed
6 weeks
May reduce the
cardiovascular risk and help
in preventing type 2
diabetes and obesity
-
↑ Bifidobacterium spp., and
Lactobacillus/Enterococcus,
showed the ability to
suppress colonic pathogen
strain Cl. histolyticum
-
May drive beneficial
alterations in gut
communities thereby
altering their collective
metabolism to contribute to
the salubrious effects on
digestive efficiency and
health
4 weaks
Significant ↓ in insulin
No significance –blood
glucos.
↑ Lactobacillus spp.
4 weeks
High anthocyanins so that it
can be used as a prebiotic
ingredient for the intestinal
microbiota in obese subjects
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Table 2. Cont.
Author
A Source of Prebiotics
Huating Li et al. (2024)
[52]
RS derived from maize
(HAM-RS2, Hi-Maize 260
resistant starch)
Jacqueline K. Palmer et.
al. (2024)
[45]
GOS
(GOSYAN® GOS)
supplementation
Xinyi Wei et al. (2020)
[44]
Hydrolyzed guar gum
(GMOS),
manno-oligosaccharide
(MOS), and
galacto-oligosaccharide
(GOS)
Nicola Johnstone et al.
(2021)
[46]
Galacto-oligosaccharides
(Biotis™ GOS, ≈7.5 g
powder~5.5 g GOS)
Participants
Type of Research
37
Randomized
placebo-controlled
crossover design trial
33
A double-blind
randomised, placebo
controlled trial
3
64
↑—increasing value. ↓—decreasing value.
In vitro fermentation
Double-blind
placebo-controlled trial
Dose
40 g/day RS
2.4 g/d
-
7.5 g/day
Period
Conclusions
8 weeks
RS-induced changes in the
gut microbiota alter the bile
acid profile, reduce
inflammation by restoring
the intestinal barrier; inhibit
lipid absorption
6 weeks
↑ Bifidobacterium but in this
instance, resulted in only
marginal effects on GI
symptoms
-
Individualized prebiotic
effects which are associated
with their chemical
structures including their
glycoside composition
4 weeks
↑ Bifidobacterium; data
presented are indicative that
GOS may be effective in
influencing the expression of
anxiety
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4.3. The Limitations of the Reviewed Studies
Disadvantages that can be observed in this review, which the authors also highlighted,
were primarily the heterogeneity of prebiotics. Namely, prebiotics are diverse and isolated
from different sources, and their chemical structure and effect can vary. This complicates
the comparison and generalization of results from different studies. Furthermore, methodological problems are highlighted, namely that research on prebiotics is often conducted on
small samples with different doses and durations, which, given the lack of standardized
protocols, can complicate the comparison of results. There is also a lack of long-term
studies that are necessary for a better understanding of their effects on health. While there
is evidence of short-term benefits, there is a lack of research tracking long-term outcomes,
such as disease prevention. Finally, every person, and therefore the composition of their gut
microbiota, is unique, so some people may benefit while others will not. It is important to
note that prebiotics have the potential to improve overall health, however, future research
should focus on addressing the aforementioned challenges in order to better understand
the effects and optimal application.
5. Conclusions
A systematic review of 30 papers on the topic of prebiotics has revealed significant
advancements in their understanding and application. The research particularly indicates
that prebiotics promote the growth of beneficial probiotic strains such as Lacticaseibacillus
rhamnosus, Lactiplantibacillus plantarum, and Bifidobacterium. In addition, innovative approaches in food production, including pasta rich in prebiotic fibers, chocolate with inulin
and stevia, and the utilization of fruit by-products, show promising results in creating
“healthier” food options. These discoveries are especially relevant in the larger context of
the global obesity epidemic, wherein prebiotics show a potential to reduce and prevent
the inflammatory markers associated with this condition. Furthermore, new doors are
opening for therapeutic interventions via discoveries of the connections between the gut
microbiome and what has been often referred to as “our second brain”—the gut itself—as
well the impact of this connection on regulating metabolism and mental health. This
work not only confirms and highlights numerous benefits of prebiotics but also aims to
stimulate the scientific community for further research to fully exploit their potential and
health benefits. In the future, scientists should investigate the long-term effects of prebiotic
supplementation and its impacts on various aspects of human health. The addition of new
types of prebiotics to the classification is anticipated to orient scientists more towards new
insights and types of prebiotic potential.
Author Contributions: Conceptualization, L.P. and M.A.; investigation, M.A. and N.K.; resources,
K.M. and K.H.; writing—original draft preparation, M.A. and L.P.; writing—review and editing, K.M.
and B.K.; supervision, K.M. and B.K. All authors have read and agreed to the published version of
the manuscript.
Funding: This research received no external funding.
Conflicts of Interest: The authors declare no conflicts of interest.
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