The objective of this study was to determine whether or not gum Arabic may serve as an efficient prebiotic for humans when they ingest it for up to four weeks. In addition, if this is the case, we need to determine the dose–effect link for any possible discoveries. Gum Arabic (EmulGold®) was dissolved in water and administered orally to healthy persons in amounts ranging from 5 to 40 grams daily. This practice lasted for a maximum of four weeks at a time. The consumption of water daily served as the control for the negative variable.
In contrast, the consumption of 10 grams of inulin functioned as the positive variable in this experiment. We used real-time PCR techniques to determine the number of bacteria present in stool samples taken at 0, 1, 2, and 4 weeks after the start of the study. In order to take into consideration any potential restrictions, questionnaires were filled out. Bacteria belonging to the genera Bacteroides, Clostridium difficile, and Enterococci were regarded to have the potential to cause damage, while bacteria belonging to the genera Bifidobacteria, and Lactobacilli were thought to have potentially beneficial effects. This difference was drawn based on the issue of whether or not these counts were already out of balance in the host or whether they afterward got out of balance. Four weeks after intake, the amounts of Bifidobacteria and Lactobacilli found in gum arabic were significantly more significant compared to the untreated negative control. The optimal dose was around 10 grams. In addition, the amounts of Bifidobacteria, Lactobacilli, and Bacteroides found in gum arabic at this dose were much higher than those found in inulin. During the investigation, we have yet to encounter any particularly challenging roadblocks. Researchers have discovered that gum arabic has prebiotic action, at the very least on par with inulin. It was found that the optimal dose was 10 grams taken once per day. This was shown to be the most effective amount.
Introduction:
The acacia tree, also known as Acacia Senegal or Acacia seyal, produces an exudate that is then dried and sold as gum arabic. The acacia tree may grow naturally in many tropical and subtropical areas throughout the whole planet, even though it is most widespread in Africa. It is a heteropolysaccharide with an appreciable amount of mol weight (about 350–850 kDa). It is found to have residues of galactose, rhamnose, arabinose, and glucuronic acid. Furthermore, it has mineral components such as calcium, potassium, and magnesium. A limit is placed on the total amount of protein, and it must not be more than 3% of the total. It is feasible to acquire concentrations of gum arabic in the water of up to forty percent without seeing a substantial change in the substance's viscosity. This is because gum arabic is insoluble in water. Because it has this function, it is an attractive candidate component that may be used in various applications, including beverages. One of the most promising applications of gum arabic is as a prebiotic. This is because gum arabic is relatively inaccessible to the various enzymes in the small intestine. Having the definition of a non-digestible food ingredient that has beneficial effects on the host by selectively stimulating the growth and activity of one or a limited number of bacteria in the colon without stimulating that of unwanted bacteria, probiotics are often referred to as prebiotics or probiotic foods.
Because of these effects, the parasite will contribute to the general well-being and health of the host. Several research has pointed to the possibility that it might be used as a component of prebiotics. Wyatt et al. explored the issue by giving 10 grams of gum arabic to one of the volunteers who participated in the study. They discovered that their intervention had increased the quantity of Bacteroides and Bifidobacterium in the sample. Cherbut et al. observed that ingesting 10 and 15 g/d for ten days boosted the numbers of both lactic acid-producing bacteria and Bifidobacteria in fecal matter samples collected from healthy human volunteers. These samples were taken from the individuals' intestines. The contents of the participants' feces were analyzed, which allowed for the determination of these findings. Research carried out in vitro by Michel et al., and May et al. yielded data that conflicted with one another on the selective development of unwanted Clostridium difficile.
Both groups employed the same research procedure. A variation could explain the former in the host that gave the feces sample; in the former example, humans provided the sample, but in the later case, pigs did so. Alternatively, the latter could be described by a difference in the host that provided the feces sample. In addition, it was found that the production of SCFA was distinct from one fiber source to the next, which resulted from the fact that the incubation procedure used different fiber sources. The potential benefits of gum arabic on gastrointestinal health are highlighted by the fact that it improves the gut membrane's function and affects the composition of SCFA when tested in vitro. Consumption of gum arabic is associated with a variety of various actions that take place in the body. One is strengthening the host's resistance in the colon, shown via experimental animal models. However, it has been postulated that this might affect the lowering of the glycemic index. The scientific community has not produced any evidence in favor of this hypothesis. Gum arabic is a molecule that, as a consequence of all of these qualities, has the potential to be fascinating concerning the functional elements of gastrointestinal health. Additionally, gum arabic has been used as a coating agent in several applications. It is good knowledge that exposure to this drug enhances the probiotic community's capacity to survive the physiological circumstances of the gut.
Because the majority of the research that was discussed before involved in vitro and animal models. There needs to be more fundamental knowledge of how gum arabic is used in man. It has not been determined whether or not the ingestion of gum arabic affects the quantitative and qualitative composition of microorganisms found in the colon flora of healthy men over an extended length of time. In order to determine gum Arabic’s prebiotic potential after it had been consumed for four weeks, researchers compared its properties to those of the well-known component inulin and used water as a negative control. Because different concentrations of gum arabic were dissolved in different amounts of water. As a result, this particular research aimed to evaluate a possible prebiotic activity shown by gum arabic. After four weeks, during which daily intake took place, a dose–effect connection should be determined if one exists.
Methods
Material
Kerry Ingredients (Cam, UK) supplied gum arabic (EmulGold®) in a spray-dried form as a water-soluble free-flowing food-grade powder. This type of gum arabic was given by Kerry Ingredients. At least 80% of the ingredients in EmulGold® are soluble fibers, and just 3% are proteins. The FDA has determined that gum arabic is GRAS. There is currently not suggested daily dosage that has been defined.
The fructo-oligosaccharide polymer inulin (Fibroline® Instant), which was graciously donated by Cosucra (Warcoing, Belgium), is generated from chicory root. It was also produced in the form of a spray-dried powder, and its normal composition includes a maximum of 10% free fructose, glucose, and saccharose, as well as at least 90% indigestible fiber. The degree of polymerization averages out to be a 9. All of the goods that were evaluated met the criteria for use in food.
Subjects and research strategy
The present study was conducted as an experiment that was randomized, double-blind, and double-controlled. A total of six separate test groups worked in parallel for the study. Before the research was carried out, the University of Maastricht's Medical Ethics Committee reviewed and approved the protocol to be used in the study. T Netherlands (MEC 05-148) followed the guidelines specified in the World Medical Association's Declaration of Helsinki to ensure its validity. The original pool of healthy volunteers, which totaled 54, was chosen based on their ability to fulfill a set of inclusion requirements, such as having a body mass index (BMI) that ranged from 19 to 27 kg/m2. Possessing normal levels of glucose and albumin in their urine, in addition to having average blood cell counts and average concentrations of glucose in the blood when the patient has not fasted. No medication is taken that affects the normal functioning of the gastrointestinal system. For example, antimicrobial drugs, avoiding functional meals, special supplements, or other products that claim to have unique pro and prebiotic qualities (yogurts, etc.). It was required that all volunteers sign the Informed Consent Form. In all, eighty-one volunteers were investigated for possible participation in the study, but only fifty-one ended up participating. At the beginning of the experiment, three volunteers decided not to continue taking part for various personal reasons and dropped out of the study.
The distribution of individuals among the different categories was decided by chance.
Experimental techniques
Everyone who agreed to participate in the research was granted a place in one of the six groups according to a Latin square layout. The entry date was used as the controlling factor for the study. The participants consumed 5, 10, 20, or 40 g of EmulGold® daily for four weeks. A daily dose of 10 grams of fibruline was administered in place of the negative control, which was water.
After dissolving in a total of 250 milliliters of water, each product was subsequently consumed somewhere between 10:00 and 10:00 in the morning. The amount of water (250 ml) that served as the control for the absence of an effect had nothing added. Throughout the study, it was stressed to the participants how important it was to maintain their everyday eating routines.
Using a standardized collection method, the volunteers were instructed to collect two fresh fecal samples ranging from approximately 10 to 15 ml in special tubes. They were also instructed to store the samples directly in the freezer or refrigerator if they did not have access to a freezer. This was performed at the start of the trial and after 1, 2, and 4 weeks had passed. After they had been given the request, they were instructed to bring the samples to the laboratory as quickly as possible.
Participants were asked to assess how well they were doing about gastrointestinal side effects at the beginning of weeks 0, 1, 2, and 4. Some of the adverse effects were aches and pains in the abdomen, intestinal bloating, gas, nausea, borborygmi, diarrhea, and constipation. After that, the ratings were as follows: 0 indicated no symptoms, 1 indicated mild, 2 indicated moderate, and three indicated severely. In addition, the frequency of stools and their consistency were analyzed and given the following ratings: 1 for watery/diarrheal, 2 for soft, 3 for average, and 4 for hard.
Analysis of microorganisms in Frozen Feces Samples
The frozen samples were collected regularly and then sent to the appropriate microbiological laboratory for analysis (Laboratorium Pro Health BV, Weert, The Netherlands). On the LightCycler® System, real-time PCR was used with 16S ribosomal DNA-targeted genus-specific primers to determine the relative abundance of Bifidobacteria, Lactobacilli, Bacteroides, C. difficile, and Enterococci. The primers that were employed showed a high PCR efficiency, ranging from 96.5 to 100.1%, and they did not demonstrate any cross-reactivity with any other types of bacteria. When extracting DNA from the different samples of feces, the QIamp® DNA Stool Micro-kit was the instrument of choice (Qiagen, Hilden, Germany). The intensity of the fluorescence generated by SYBR Green I was measured to determine the amount of PCR product created after each amplification cycle. Standard curves were generated for each bacterial group, or species in the case of C. difficile, by plotting threshold cycles obtained from real-time PCR analyses of various culture dilutions of bacterial strains representative for the groups. These bacterial strains were used to ensure that the standard curves were accurate.
Following that step, each bacterial strain's relative abundance was calculated using these standard curves. Plate counting was used to determine the number of bacteria, which enabled a direct comparison between the threshold cycle and the ten logarithms of the number of bacteria. The results were calculated based on the precise amount of excrement collected, and they were provided as the log10 number of bacteria per gram of wet feces.
Results
Compliance shown by the volunteers.
The information on the foundation characteristics of the distinct groups is shown in Table 2. The different groups did not vary from one another in any of these traits in any way that could be considered statistically significant. The amount of the testing product consumed by the subjects throughout the research was used to determine whether or not they complied with the protocol. Compliance was determined to be extremely high, ranging from 97.0 to 99.6%.
Table 2: Baseline demographic parameters of the research population that were employed for statistical analysis*
Evaluation of the bacterial population after four weeks of intervention
Table 3 lists the exact data of the different kinds of bacteria that existed at the beginning and the period's conclusion. The four groups of participants did not vary significantly in terms of the initial amount of the different bacteria found in their mouths. The difference between the 10-logarithmic figure at the beginning of the study and that number after four weeks was compared and analyzed. A discernible shift in these bacterial populations was seen only in the participants who had previously taken gum arabic. It was shown that the levels of Bifidobacteria, Lactobacilli, and Bacteroides significantly increased after a dosage of 10 grams. At a dosage of 20 grams, the number of Lactobacilli decreased, whereas, at a dosage of 40 grams, the number of Bacteroides decreased.
Table 3 presents the ten logarithmic counts of bacteria in each of the different groups at the beginning of the research (week 0) and the investigation's conclusion (week 4).
Bifidobacteria spp.
The dosage had an impact that was proportional to the change in the number of Bifidobacteria that were found in the feces (Table 4). The rise in the populations of Bifidobacteria in patients who had ingested 10 grams of gum arabic was considerably (P 0.001) larger than in those who had 0 grams of gum arabic (water, the negative control): a differential in an outgrowth of around 40 times. In addition, this rise was substantially (P 0.05) more extraordinary for gum arabic at a dosage of 10 g than it was for inulin: a difference nearly 10 times larger. More significant quantities of gum arabic have not resulted in a more remarkable impact.
The regression model determined that the ideal dosage, in terms of the most significant possible number of bacteria, was somewhere around 10 grams of gum arabic.
Lactobacilli spp.
After taking 10 grams of gum arabic per day for four weeks, the rise in the number of lactobacilli was considerably (P 0.05) larger than that which was found at 0 grams of gum arabic (water, the negative control): the difference in numbers was almost six times greater. Interestingly, eating 10 grams of gum arabic resulted in a considerably (P 0.03) more significant rise in numbers than that seen with 10 grams of inulin: about a sevenfold increase (Table 4).
It was determined that a dosage of around 5–10 g of EmulGold® provides the best results regarding the number of lactobacilli.
Bacteroides spp.
Except at a dosage of 10 grams, when an increase of nearly twofold was discovered, no discernible changes were identified in the number of Bacteroides between the volunteers who consumed gum arabic for four weeks and the participants who consumed water. Interestingly, the consumption of 10 grams of gum arabic resulted in a considerably (P 0.01) more significant rise in counts than 10 grams of inulin (about 2.5 times; Table 4).
At dosages of 5 and 10 g of gum arabic, there were discovered to be quite considerable alterations in the populations of Bacteroides. Similar to what was seen with the Lactobacilli, the quantity of Bacteroides decreased with increasing dosages, and the difference between the two groups was statistically significant (P 0.001).
Clostridium difficile
There was no discernible difference in the total number of C. difficile cases between the groups, either at the beginning or after four weeks (Table 4). However, the variance in numbers at each time point was very considerable. This was notably the case when increasing dosages of gum arabic were consumed, but it was also when the intervention began. Not only were there no discernible changes found between water and the different concentrations of gum arabic, but there were also no discernible differences found between inulin and gum arabic. After an intervention lasting for four weeks, there was no dose-dependence in the alterations that occurred in the quantities of bacteria.
Enterococci spp.
A drop in bacterial counts was seen over the intervention period of 4 weeks, although it was not statistically significant. This was particularly the case in the water group (Table 4). There were no discernible differences in the number of germs across the different test groups that could be classified as statistically significant. There was no indication of a possible dose-dependency in the result when gum arabic was used.
Possible adverse effects resulting from the use of gum arabic for four weeks
The following gastrointestinal symptoms were evaluated using a questionnaire: the consistency of feces; stomach discomfort; nausea; borborygmi; colic pains; bloating; flatulence; diarrhea; and constipation. Stool regularity and stool texture were observed throughout the trial. Even at the maximum dosage of 40 g/day, all test participants could consume gum arabic for four weeks without any adverse effects. Compared to participants who ingested either the positive or negative control, there was no significant difference in the subjects' reported changes in gastrointestinal symptoms over the intervention period. These changes were modest. The fluctuation in the severity of diarrhea, which is shown in Figure 1, serves as an illustration of the discomfort that patients experience. People who reported having diarrhea during the initial measurement are represented by numbers that are lower than 0. The intake of all substances and dosages tested for four weeks did not result in a significant increase in the number of cases of diarrhea.
Discussion
The current research findings make it abundantly evident that ing gum arabic (EmulGold®) at a dosage of 10 g per day for four weeks significantly reduced the risk of developing ovarian cancer, which has a more significant number of beneficial bacteria, such as lactobacilli and Bifidobacteria, than water (negative control). It was discovered that the consumption of gum arabic resulted in considerably more significant quantities of Bifidobacteria, Lactobacilli, and Bacteroides when compared to the consumption of 10 grams of inulin, which served as a positive control. It was not substantially different from the impact of water or inulin on the populations of other bacteria. It has been shown that gum arabic may be regarded as a prebiotic fiber functioning at least as excellent as inulin. This is because gum arabic produces a more significant number of helpful bacteria without stimulating any undesirable bacteria.
The prebiotic debate centers on several essential questions, one of which is whether the floral makeup is in a state of equilibrium. The number of bacteria in the colon that are out of balance is related to whether the bacteria are helpful to the host. Species, including Bifidobacteria and Lactobacilli, are widely known for their health-improving properties, such as a rise in the resistance to colonization or the generation of SCFA. Because of this, they are often utilized as probiotics. In addition, each group of bacteria in the colon has its unique species, which may or may not be advantageous to the body. One such example is the group known as Bacteroides. The fact that gum arabic caused an increase in the log10 number of Bacteroides was one of the most noteworthy findings of this investigation. Like any other genus of bacteria, Bacteroides are made up of both helpful and harmful organisms to human health. Some recent publications discuss the positive effects of members of this group on the health of the stomach. Therefore, a study into the role of individual individuals within bacterial groupings might uncover the health-improving qualities.
According to the most recent findings from research on bacterial proliferation, the optimal amount of gum arabic to produce prebiotic action is between 5 and 10 grams. This amount was distributed weekly over four weeks. It is an interesting observation that the numbers decreased as the dose grew. When the amount of Bifidobacteria is increased to 10 grams, the number of Bifidobacteria hits a plateau level and then stays at the same level there. According to the findings of this research, Bifidobacteria can metabolize gum arabic at concentrations of up to 40 grams. However, other bacteria have more difficulty doing so at higher concentrations. There may be competition for the same substrate between more than one organism at large doses. Certain bacterial strains are more able than others to break through the substrate they are growing on.
Consequently, fewer resources will be offered to those individuals whose qualities have been identified via this study as having existed in the past. The enzyme systems found within Bifidobacteria are more adapted to digest gum arabic than the enzyme systems found within other kinds of microbes. There is also the possibility of the composition of the various layers of microflora that are present in the gastrointestinal tract, as well as the subsequent utilization of substrate that can degrade the prebiotic at higher doses. In addition, there is the possibility that microflora is present in the gastrointestinal tract. In this case, we would be dealing with flora connected with the surface, as opposed to flora linked with the mucosa. Because they have easier access to the chemical when it is present in higher concentrations, bacteria already present in the mucosa layer may be able to degrade the chemicals to a more significant extent than they can when it is present at lower levels. One may form a hypothesis based on this. It is not yet known to what extent the creation of specific compounds by particular gut bacteria as a result of the fermentation of gum arabic impacts the growth of other bacteria; this must be determined.
This study used a real-time PCR test to quantify the number of relevant bacteria that were present in the fecal slurry, which represented the bacteria that are located in the lumen of the gut. This approach has several drawbacks, including the fact that it is dependent not only on whether or not there is a direct link in the quantity and composition of bacteria between a sample of fecal slurry and the lumen of the gut but also on the validity of the primers that were utilized: I would be interested in finding out how accurate the binding of the used amino acid array to the specific DNA in the feces is.
There was a significant association between DNA and threshold cycles and between bacterially spiked fecal samples and signals, as was discovered and reported on. In addition, an internal validation was carried out on this study using several sample analyses. This was done in order to support the findings of the inquiry. In addition, feces samples were spiked with a known inoculum, and the results of this experiment were proved to be statistically trustworthy.
Not only did the total number of bacteria undergo considerable shifts throughout the study, but they also did so from the very start. This was an essential part of the task that was carried out. There was a significant amount of individual variation among the C. difficile; in fact, it was more than five log numbers. This was notably the case for those willing participants who ingested 10 grams of gum arabic. It has been shown in the past that there is a clear connection between the total number of bacteria present in the feces before the beginning of the intervention and the net increase resulting from the intervention. The findings of the most recent inquiry substantiated the veracity of this assertion (Fig. 2). Figure 2 presents data that demonstrate a statistically significant inverse connection (P.0001) between the log10 number of C. difficile that was present at the beginning of the study and the rate of change in numbers after ingestion of gum arabic for four weeks. This connection is shown between the log10 number of C. difficile that was present and the rate of change in numbers. When there are few bacteria at the beginning of the experiment, there is little area for the population to decline, and the numbers can only grow.
On the other hand, things take a completely different turn when there are many bacteria. There was no effort made at any point at the beginning of the study endeavor to count the number of germs in the participants' feces. However, since we verified the diets of all participants before beginning the study, we could select the volunteers who consumed yogurt consistently and were thus likely to have a high amount of lactic acid bacteria in their intestines. The intervention could result in a shift in the overall number of bacteria over time. In order to lessen the probability of their being biased in the research results, the criteria for inclusion should be revised so that it is possible to eliminate individuals whose numbers were either extraordinarily low or excessively high before the study started.
At this point, just a few studies have shown proof of the relatively long-lasting prebiotic advantages that gums, in general, have on people. The physiological effects that other gums display after intake are analogous to Arabic Gum prebiotic qualities, which are shown to benefit human health. Over 21 days, supplementation with partially hydrolyzed guar gum and fructo-oligosaccharides at doses of 6.6 and 3.4 g/d, respectively, resulted in a substantial increase in the number of Bifidobacteria. On the other hand, there was no change in the number of Lactobacilli, Bacteroides, Clostridia, or Enterococci. In addition, partially hydrolyzed guar appears to aid in ameliorating symptoms associated with inflammatory bowel syndrome. In vitro research conducted with bacteria taken from the human colon revealed that gum arabic is rapidly consumed by these microorganisms more than any other substance investigated. It is common knowledge that the application of fibers works not only by increasing the quantities of bacteria but also by activating the physiological changes of bacteria and, as a result, by modifying the release of certain substances. This is one of the many ways the application of fibers achieves its desired effect. For instance, lowering the concentrations of SCFA and various other chemicals that are known to have the potential to cause cancer.
The findings of this research indicate that gum arabic (EmulGold®) has prebiotic activity within a dosage range comparable to or lower than inulin. This was determined by the quantitative growth of bacteria in stool samples. This chemical will be an exciting component for the functional food market because of the functional qualities it has inside food matrices. The question of whether or not it will display additional physiologically significant features will have to be answered by future research.