Nursing Nutrition and Dietetics

Microbial Metabolites in Food Biotechnology

Description

This cluster of papers explores the role of fructans, such as inulin and oligofructose, in nutrition and health. It covers their impact on gut microbiota, calcium absorption, immune modulation, and their potential as prebiotics and dietary fiber. The papers also discuss the health benefits and applications of fructans in functional foods.

Keywords

Fructans; Inulin; Oligofructose; Prebiotics; Health Benefits; Dietary Fiber; Gut Microbiota; Calcium Absorption; Immunomodulation; Functional Food

Interest has been recently rekindled in short chain fatty acids (SCFAs) with the emergence of prebiotics and probiotics aimed at improving colonic and systemic health. Dietary carbohydrates, specifically resistant starches … Interest has been recently rekindled in short chain fatty acids (SCFAs) with the emergence of prebiotics and probiotics aimed at improving colonic and systemic health. Dietary carbohydrates, specifically resistant starches and dietary fiber, are substrates for fermentation that produce SCFAs, primarily acetate, propionate, and butyrate, as end products. The rate and amount of SCFA production depends on the species and amounts of microflora present in the colon, the substrate source and gut transit time. SCFAs are readily absorbed. Butyrate is the major energy source for colonocytes. Propionate is largely taken up by the liver. Acetate enters the peripheral circulation to be metabolized by peripheral tissues. Specific SCFA may reduce the risk of developing gastrointestinal disorders, cancer, and cardiovascular disease. Acetate is the principal SCFA in the colon, and after absorption it has been shown to increase cholesterol synthesis. However, propionate, a gluconeogenerator, has been shown to inhibit cholesterol synthesis. Therefore, substrates that can decrease the acetate: propionate ratio may reduce serum lipids and possibly cardiovascular disease risk. Butyrate has been studied for its role in nourishing the colonic mucosa and in the prevention of cancer of the colon, by promoting cell differentiation, cell-cycle arrest and apoptosis of transformed colonocytes; inhibiting the enzyme histone deacetylase and decreasing the transformation of primary to secondary bile acids as a result of colonic acidification. Therefore, a greater increase in SCFA production and potentially a greater delivery of SCFA, specifically butyrate, to the distal colon may result in a protective effect. Butyrate irrigation (enema) has also been suggested in the treatment of colitis. More human studies are now needed, especially, given the diverse nature of carbohydrate substrates and the SCFA patterns resulting from their fermentation. Short-term and long-term human studies are particularly required on SCFAs in relation to markers of cancer risk. These studies will be key to the success of dietary recommendations to maximize colonic disease prevention.
Flavonoid and isoflavonoid glycosides are common dietary phenolics which may be absorbed from the small intestine of humans. The ability of cell‐free extracts from human small intestine and liver to … Flavonoid and isoflavonoid glycosides are common dietary phenolics which may be absorbed from the small intestine of humans. The ability of cell‐free extracts from human small intestine and liver to deglycosylate various (iso)flavonoid glycosides was investigated. Quercetin 4′‐glucoside, naringenin 7‐glucoside, apigenin 7‐glucoside, genistein 7‐glucoside and daidzein 7‐glucoside were rapidly deglycosylated by both tissue extracts, whereas quercetin 3,4′‐diglucoside, quercetin 3‐glucoside, kaempferol 3‐glucoside, quercetin 3‐rhamnoglucoside and naringenin 7‐rhamnoglucoside remained unchanged. The K m for hydrolysis of quercetin 4′‐glucoside and genistein 7‐glucoside was ∼32±12 and ∼14±3 μM in both tissues respectively. The enzymatic activity of the cell‐free extracts exhibits similar properties to the cytosolic broad‐specificity β‐glucosidase previously described in mammals.
Prebiotics are food ingredients that improve health by modulating the colonic microbiota. The bifidogenic effect of the prebiotic inulin is well established; however, it remains unclear which species of Bifidobacterium … Prebiotics are food ingredients that improve health by modulating the colonic microbiota. The bifidogenic effect of the prebiotic inulin is well established; however, it remains unclear which species of Bifidobacterium are stimulated in vivo and whether bacterial groups other than lactic acid bacteria are affected by inulin consumption. Changes in the faecal microbiota composition were examined by real-time PCR in twelve human volunteers after ingestion of inulin (10 g/d) for a 16-d period in comparison with a control period without any supplement intake. The prevalence of most bacterial groups examined did not change after inulin intake, although the low G+C % Gram-positive species Faecalibacterium prausnitzii exhibited a significant increase (10·3 % for control period v. 14·5 % during inulin intake, P = 0·019). The composition of the genus Bifidobacterium was studied in four of the volunteers by clone library analysis. Between three and five Bifidobacterium spp. were found in each volunteer. Bifidobacterium adolescentis and Bifidobacterium longum were present in all volunteers, and Bifidobacterium pseudocatenulatum , Bifidobacterium animalis , Bifidobacterium bifidum and Bifidobacterium dentium were also detected. Real-time PCR was employed to quantify the four most prevalent Bifidobacterium spp., B. adolescentis , B. longum , B. pseudocatenulatum and B. bifidum , in ten volunteers carrying detectable levels of bifidobacteria. B. adolescentis showed the strongest response to inulin consumption, increasing from 0·89 to 3·9 % of the total microbiota ( P = 0·001). B. bifidum was increased from 0·22 to 0·63 % ( P < 0·001) for the five volunteers for whom this species was present.
In November 2008, a group of scientists met at the 6th Meeting of the International Scientific Association of Probiotics and Prebiotics (ISAPP) in London, Ontario, Canada, to discuss the functionality … In November 2008, a group of scientists met at the 6th Meeting of the International Scientific Association of Probiotics and Prebiotics (ISAPP) in London, Ontario, Canada, to discuss the functionality of prebiotics. As a result of this, it was concluded that the prebiotic field is currently dominated by gastrointestinal events. However, in the future, it may be the case that other mixed microbial ecosystems may be modulated by a prebiotic approach, such as the oral cavity, skin and the urogenital tract. Therefore, a decision was taken to build upon the current prebiotic status and define a niche for ‘dietary prebiotics’. This review is co-authored by the working group of ISAPP scientists and sets the background for defining a dietary prebiotic as ‘‘a selectively fermented ingredient that results in specific changes in the composition and/or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health’’.
Inulin is a generic term to cover all β(2 → 1) linear fructans. Chicory inulin is a linear β(2 → 1) fructan (degree of polymerisation (DP) 2 to 60; DP … Inulin is a generic term to cover all β(2 → 1) linear fructans. Chicory inulin is a linear β(2 → 1) fructan (degree of polymerisation (DP) 2 to 60; DP av = 12), its partial enzymatic hydrolysis product is oligofructose (DP 2 to 8; DP av = 4), and by applying specific separation technologies a long-chain inulin knownas inulin HP (DP 10 to 60; DP av = 25) can be produced. Finally, a specific product known as oligofructose-enriched inulin is obtained by combining chicorylong-chain inulin and oligofructose. Because of the b-configuration of the anomeric C2 in their fructose monomers, inulin-type fructans resist hydrolysis byintestinal digestive enzymes, they classify as ‘non-digestible’ carbohydrates, and they are dietary fibres. By increasing faecal biomass and water content ofthe stools, they improve bowel habits, but they have characteristic features different from other fibres. They affect gastrointestinal functions not because oftheir physico-chemical properties but rather because of their biochemical and physiological attributes. In the colon, they are rapidly fermented to produceSCFA that are good candidates to explain some of the systemic effects of inulin-type fructans. Fermentation of inulin-type fructans in the large bowel is aselective process; bifidobacteria (and possibly a few other genera) are preferentially stimulated to grow, thus causing significant changes in the compositionof the gut microflora by increasing the number of potentially health-promoting bacteria and reducing the number of potentially harmful species. Both oligofructosead inulin are prebiotic. They also induce changes in colonic epithelium stimulating proliferation in the crypts, increasing the concentration ofpolyamines, changing the profile of mucins, and modulating endocrine as well as immune functions. From a nutrition labelling perspective, inulin-typefructans are not only prebiotic dietary fibres; they are also low-calorie carbohydrates [6·3 kJ/g (1·5 kcal/g)]. Supported by the results of a large numberof animal studies and human nutrition intervention trials, the claim ‘inulin-type fructans enhance calcium and magnesium absorption‘ is scientifically substantiated, but different inulin-type fructans have probably a different efficacy (in terms of effective daily dose), the most active product being the oligofructose-enriched inulin. A series of animal studies demonstrate that inulin-type fructans affect the metabolism of lipids primarily by decreasingtriglyceridaemia because of a reduction in the number of plasma VLDL particles. The human data largely confirm the animal experiments. They demonstratemainly a reduction in triglyceridaemia and only a relatively slight decrease in cholesterolaemia mostly in (slightly) hypertriglyceridaemic conditions. Inulinappears thus eligible for an enhanced function claim related to normalization of blood triacylglycerols. A large number of animal data convincingly showthat inulin-type fructans reduce the risk of colon carcinogenesis and nutrition intervention trials are now performed to test that hypothesis in human subjectsknown to be at risk for polyps and cancer development in the large bowel.
SUMMARY: An improved growth medium for lactobacilli is described. It supports good growth of lactobacilli generally and also is particularly useful for a number of fastidious strains which grow only … SUMMARY: An improved growth medium for lactobacilli is described. It supports good growth of lactobacilli generally and also is particularly useful for a number of fastidious strains which grow only poorly in other general media. In addition, tomato juice, a highly variable material, is not required. In a slightly modified form, it can also be used as a basal medium for fermentation tests.
Dietary carbohydrates have the potential to influence diverse functional groups of bacteria within the human large intestine. Of 12 Bifidobacterium strains of human gut origin from seven species tested, four … Dietary carbohydrates have the potential to influence diverse functional groups of bacteria within the human large intestine. Of 12 Bifidobacterium strains of human gut origin from seven species tested, four grew in pure culture on starch and nine on fructo-oligosaccharides. The potential for metabolic cross-feeding between Bifidobacterium adolescentis and lactate-utilizing, butyrate-producing Firmicute bacteria related to Eubacterium hallii and Anaerostipes caccae was investigated in vitro. E. hallii L2-7 and A. caccae L1-92 failed to grow on starch in pure culture, but in coculture with B. adolescentis L2-32 butyrate was formed, indicating cross-feeding of metabolites to the lactate utilizers. Studies with [(13)C]lactate confirmed carbon flow from lactate, via acetyl coenzyme A, to butyrate both in pure cultures of E. hallii and in cocultures with B. adolescentis. Similar results were obtained in cocultures involving B. adolescentis DSM 20083 with fructo-oligosaccharides as the substrate. Butyrate formation was also stimulated, however, in cocultures of B. adolescentis L2-32 grown on starch or fructo-oligosaccharides with Roseburia sp. strain A2-183, which produces butyrate but does not utilize lactate. This is probably a consequence of the release by B. adolescentis of oligosaccharides that are available to Roseburia sp. strain A2-183. We conclude that two distinct mechanisms of metabolic cross-feeding between B. adolescentis and butyrate-forming bacteria may operate in gut ecosystems, one due to consumption of fermentation end products (lactate and acetate) and the other due to cross-feeding of partial breakdown products from complex substrates.
The lactate dehydrogenase (LDH) isoenzyme spectrum enables optimally controlling T cell glycolysis and differentiation, The lactate dehydrogenase (LDH) isoenzyme spectrum enables optimally controlling T cell glycolysis and differentiation,
Abstract The task of inventing and developing active ingredients with useful biological activities requires a search for novel chemical substructures. This process may trigger the discovery of whole classes of … Abstract The task of inventing and developing active ingredients with useful biological activities requires a search for novel chemical substructures. This process may trigger the discovery of whole classes of chemicals of potential commercial interest. Similar biological effects can often be achieved by completely different compounds. However, compounds within a given structural family may exhibit quite different biological activities depending on their interactions with different intracellular proteins like enzymes or receptors. By varying the functional groups and structural elements of a lead compound, its interaction with the active site of the target protein, as well as its physicochemical, pharmacokinetic, and dynamic properties can be improved. In this context, the introduction of fluorine into active ingredients has become an important concept in the quest for a modern crop protection product with optimal efficacy, environmental safety, user friendliness, and economic viability. Fluorinated organic compounds represent an important and growing family of commercial agrochemicals. A number of recently developed agrochemical candidates represent novel classes of chemical compounds with new modes of action; several of these compounds contain new fluorinated substituents. However, the complex structure–activity relationships associated with biologically active molecules mean that the introduction of fluorine can lead to either an increase or a decrease in the efficacy of a compound depending on its changed mode of action, physicochemical properties, target interaction, or metabolic susceptibility and transformation. Therefore, it is still difficult to predict the sites in a molecule at which fluorine substitution will result in optimal desired effects.
The health benefits of dietary fiber have long been appreciated. Higher intakes of dietary fiber are linked to less cardiovascular disease and fiber plays a role in gut health, with … The health benefits of dietary fiber have long been appreciated. Higher intakes of dietary fiber are linked to less cardiovascular disease and fiber plays a role in gut health, with many effective laxatives actually isolated fiber sources. Higher intakes of fiber are linked to lower body weights. Only polysaccharides were included in dietary fiber originally, but more recent definitions have included oligosaccharides as dietary fiber, not based on their chemical measurement as dietary fiber by the accepted total dietary fiber (TDF) method, but on their physiological effects. Inulin, fructo-oligosaccharides, and other oligosaccharides are included as fiber in food labels in the US. Additionally, oligosaccharides are the best known “prebiotics”, “a selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microflora that confers benefits upon host well-bring and health.” To date, all known and suspected prebiotics are carbohydrate compounds, primarily oligosaccharides, known to resist digestion in the human small intestine and reach the colon where they are fermented by the gut microflora. Studies have provided evidence that inulin and oligofructose (OF), lactulose, and resistant starch (RS) meet all aspects of the definition, including the stimulation of Bifidobacterium, a beneficial bacterial genus. Other isolated carbohydrates and carbohydrate-containing foods, including galactooligosaccharides (GOS), transgalactooligosaccharides (TOS), polydextrose, wheat dextrin, acacia gum, psyllium, banana, whole grain wheat, and whole grain corn also have prebiotic effects.
Several species of protozoa, during their logarithmic phases of growth, produce substances that prolong the logarithmic phase in other species. The effect is not as striking as the inhibition of … Several species of protozoa, during their logarithmic phases of growth, produce substances that prolong the logarithmic phase in other species. The effect is not as striking as the inhibition of growth caused by antibiotics, but a consistent 50-percent increase in growth has been obtained with Tetrahymena pyriformis in response to a factor produced by Colpidium campylum.
Most studies involving prebiotic oligosaccharides have been carried out using inulin and its fructo-oligosaccharide (FOS) derivatives, together with various forms of galacto-oligosaccharides (GOS). Although many intestinal bacteria are able to … Most studies involving prebiotic oligosaccharides have been carried out using inulin and its fructo-oligosaccharide (FOS) derivatives, together with various forms of galacto-oligosaccharides (GOS). Although many intestinal bacteria are able to grow on these carbohydrates, most investigations have demonstrated that the growth of bifidobacteria, and to a lesser degree lactobacilli, is particularly favoured. Because of their safety, stability, organoleptic properties, resistance to digestion in the upper bowel and fermentability in the colon, as well as their abilities to promote the growth of beneficial bacteria in the gut, these prebiotics are being increasingly incorporated into the Western diet. Inulin-derived oligosaccharides and GOS are mildly laxative, but can result in flatulence and osmotic diarrhoea if taken in large amounts. However, their effects on large bowel habit are relatively minor. Although the literature dealing with the health significance of prebiotics is not as extensive as that concerning probiotics, considerable evidence has accrued showing that consumption of GOS and FOS can have significant health benefits, particularly in relation to their putative anti-cancer properties, influence on mineral absorption, lipid metabolism, and anti-inflammatory and other immune effects such as atopic disease. In many instances, prebiotics seem to be more effective when used as part of a synbiotic combination.
Two hundred forty male Avian Farms broiler chicks, 1 d of age, were randomly allocated to four treatments, each of which had five pens of 12 chicks per pen. The … Two hundred forty male Avian Farms broiler chicks, 1 d of age, were randomly allocated to four treatments, each of which had five pens of 12 chicks per pen. The chicks were used to investigate the effects of fructooligosaccharide (FOS) on digestive enzyme activities and intestinal microflora and morphology. The chicks received the same basal diet based on corn-soybean meal, and FOS was added to the basal diet at 0, 2.0, 4.0, and 8.0 g/kg diet at the expense of corn. Addition of 4.0 g/kg FOS to the basal diet significantly increased average daily gain of broilers. The feed-to-gain ratios were significantly decreased for the birds fed diets with 2.0 and 4.0 g/kg FOS versus the control. Addition of 4.0 g/kg FOS enhanced the growth of Bifidobacterium and Lactobacillus, but inhibited Escherichia coli in the small intestinal and cecal digesta. Supplementation of 2.0 or 4.0 g/kg FOS to chicks significantly improved the activities of amylase compared to the control (12.80 or 14.75 vs. 8.42 Somogyi units). A significant increase in the activities of total protease was observed in 4.0 g/kg FOS-treated birds versus controls (83.91 vs. 65.97 units). Morphology data for the duodenum, jejunum, and ileum showed no significant differences for villus height, crypt depth, or microvillus height at the duodenum. By contrast, addition of 4.0 g/kg FOS significantly increased ileal villus height, jejunal and ileal microvillus height, and villus-height-to-crypt-depth ratios at the jejunum and ileum and decreased crypt depth at the jejunum and ileum. However, addition of 8.0 g/kg FOS had no significant effect on growth performance, digestive enzyme activities, intestinal microflora, or morphology.
Bacteria that colonize the mammalian intestine collectively possess a far larger repertoire of degradative enzymes and metabolic capabilities than their hosts. Microbial fermentation of complex non-digestible dietary carbohydrates and host–derived … Bacteria that colonize the mammalian intestine collectively possess a far larger repertoire of degradative enzymes and metabolic capabilities than their hosts. Microbial fermentation of complex non-digestible dietary carbohydrates and host–derived glycans in the human intestine has important consequences for health. Certain dominant species, notably among the Bacteroidetes, are known to possess very large numbers of genes that encode carbohydrate active enzymes and can switch readily between different energy sources in the gut depending on availability. Nevertheless, more nutritionally specialized bacteria appear to play critical roles in the community by initiating the degradation of complex substrates such as plant cell walls, starch particles and mucin. Examples are emerging from the Firmicutes, Actinobacteria and Verrucomicrobium phyla, but more information is needed on these little studied groups. The impact of dietary carbohydrates, including prebiotics, on human health requires understanding of the complex relationship between diet composition, the gut microbiota and metabolic outputs.
Prebiotics are non-digestible (by the host) food ingredients that have a beneficial effect through their selective metabolism in the intestinal tract. Key to this is the specificity of microbial changes. … Prebiotics are non-digestible (by the host) food ingredients that have a beneficial effect through their selective metabolism in the intestinal tract. Key to this is the specificity of microbial changes. The present paper reviews the concept in terms of three criteria: (a) resistance to gastric acidity, hydrolysis by mammalian enzymes and gastrointestinal absorption; (b) fermentation by intestinal microflora; (c) selective stimulation of the growth and/or activity of intestinal bacteria associated with health and wellbeing. The conclusion is that prebiotics that currently fulfil these three criteria are fructo-oligosaccharides, galacto-oligosaccharides and lactulose, although promise does exist with several other dietary carbohydrates. Given the range of food vehicles that may be fortified by prebiotics, their ability to confer positive microflora changes and the health aspects that may accrue, it is important that robust technologies to assay functionality are used. This would include a molecular-based approach to determine flora changes. The future use of prebiotics may allow species-level changes in the microbiota, an extrapolation into genera other than the bifidobacteria and lactobacilli, and allow preferential use in disease-prone areas of the body.
Butyrate is a natural substance present in biological liquids and tissues. The present paper aims to give an update on the biological role of butyrate in mammals, when it is … Butyrate is a natural substance present in biological liquids and tissues. The present paper aims to give an update on the biological role of butyrate in mammals, when it is naturally produced by the gastrointestinal microbiota or orally ingested as a feed additive. Recent data concerning butyrate production delivery as well as absorption by the colonocytes are reported. Butyrate cannot be detected in the peripheral blood, which indicates fast metabolism in the gut wall and/or in the liver. In physiological conditions, the increase in performance in animals could be explained by the increased nutrient digestibility, the stimulation of the digestive enzyme secretions, a modification of intestinal luminal microbiota and an improvement of the epithelial integrity and defence systems. In the digestive tract, butyrate can act directly (upper gastrointestinal tract or hindgut) or indirectly (small intestine) on tissue development and repair. Direct trophic effects have been demonstrated mainly by cell proliferation studies, indicating a faster renewal of necrotic areas. Indirect actions of butyrate are believed to involve the hormono-neuro-immuno system. Butyrate has also been implicated in down-regulation of bacteria virulence, both by direct effects on virulence gene expression and by acting on cell proliferation of the host cells. In animal production, butyrate is a helpful feed additive, especially when ingested soon after birth, as it enhances performance and controls gut health disorders caused by bacterial pathogens. Such effects could be considered for new applications in human nutrition.
A procedure is described for the measurement and characterisation of total non-starch polysaccharides in plant foods by gas-liquid chromatography of individual sugars as alditol acetates. Starch, including that resistant to … A procedure is described for the measurement and characterisation of total non-starch polysaccharides in plant foods by gas-liquid chromatography of individual sugars as alditol acetates. Starch, including that resistant to gelatinisation in boiling water, is dispersed with dimethyl sulphoxide and then hydrolysed with α-amylase and pullulanase. Starch-free material is hydrolysed with sulphuric acid and the released neutral sugars are measured as alditol acetates by using N-methylimidazole in order to catalyse the acetylation. Uronic acids are measured by a spectrophotometric method. The procedure is a modification of a more comprehensive method reported previously. It is relatively rapid and simple compared with gravimetric procedures, is applicable to a wide range of foodstuffs, can be carried out without special expertise in carbohydrate chemistry and is suitable for routine laboratory use.
Abstract The gut is an obvious target for the development of functional foods, acting as it does as the interface between diet and the metabolic events which sustain life. The … Abstract The gut is an obvious target for the development of functional foods, acting as it does as the interface between diet and the metabolic events which sustain life. The key processes in digestive physiology which can be regulated by modifying diet are satiety, the rate and extent of macronutrient breakdown and absorption from the small bowel, sterol metabolism, the colonic microflora, fermentation, mucosal function and bowel habit, and the gut immune system. The intestinal microflora is the main focus of many current functional foods. Probiotics are foods which contain live bacteria which are beneficial to health whilst prebiotics, such as certain non-digestible oligosaccharides which selectively stimulate the growth of bifidobacteria in the colon, are already on the market. Their claimed benefits are to alleviate lactose maldigestion, increase resistance to invasion by pathogenic species of bacteria in the gut, stimulate the immune system and possibly protect against cancer. There are very few reports of well-designed human intervention studies with prebiotics as yet. Certain probiotic species have been shown to shorten the duration of rotavirus diarrhoea in children but much more work is needed on the mechanism of immunomodulation and of competitive exclusion and microflora modification. The develop-ment of functional foods for the gut is in its infancy and will be successful only if more fundamental research is done on digestive physiology, the gut microflora, immune system and mucosal function.
One of the key features of plants is their ability to reduce carbon dioxide in the presence of sunlight and water to sugars, and the subsequent transport of assimilated carbon … One of the key features of plants is their ability to reduce carbon dioxide in the presence of sunlight and water to sugars, and the subsequent transport of assimilated carbon to the nonphotosynthetic tissues (sink tissues). In most plants, the transported sugar is Suc, a nonreducing disaccharide,
<h3>Objective</h3> To highlight the contribution of the gut microbiota to the modulation of host metabolism by dietary inulin-type fructans (ITF prebiotics) in obese women. <h3>Methods</h3> A double blind, placebo controlled, … <h3>Objective</h3> To highlight the contribution of the gut microbiota to the modulation of host metabolism by dietary inulin-type fructans (ITF prebiotics) in obese women. <h3>Methods</h3> A double blind, placebo controlled, intervention study was performed with 30 obese women treated with ITF prebiotics (inulin/oligofructose 50/50 mix; n=15) or placebo (maltodextrin; n=15) for 3 months (16 g/day). Blood, faeces and urine sampling, oral glucose tolerance test, homeostasis model assessment and impedancemetry were performed before and after treatment. The gut microbial composition in faeces was analysed by phylogenetic microarray and qPCR analysis of 16S rDNA. Plasma and urine metabolic profiles were analysed by <sup>1</sup>H-NMR spectroscopy. <h3>Results</h3> Treatment with ITF prebiotics, but not the placebo, led to an increase in <i>Bifidobacterium</i> and <i>Faecalibacterium prausnitzii</i>; both bacteria negatively correlated with serum lipopolysaccharide levels. ITF prebiotics also decreased <i>Bacteroides intestinalis</i>, <i>Bacteroides vulgatus</i> and <i>Propionibacterium</i>, an effect associated with a slight decrease in fat mass and with plasma lactate and phosphatidylcholine levels. No clear treatment clustering could be detected for gut microbial analysis or plasma and urine metabolomic profile analyses. However, ITF prebiotics led to subtle changes in the gut microbiota that may importantly impact on several key metabolites implicated in obesity and/or diabetes. <h3>Conclusions</h3> ITF prebiotics selectively changed the gut microbiota composition in obese women, leading to modest changes in host metabolism, as suggested by the correlation between some bacterial species and metabolic endotoxaemia or metabolomic signatures.
Abstract The classic definitions of inulin and oligofructose are constructively criticized. It is observed that inulin cannot unequivocally be described as a polydisperse 1‐kestose‐based (GFn) β(2 ? 1) linear fructan … Abstract The classic definitions of inulin and oligofructose are constructively criticized. It is observed that inulin cannot unequivocally be described as a polydisperse 1‐kestose‐based (GFn) β(2 ? 1) linear fructan chain, but that inulin always contains small amounts of Fm and branched molecules. This review article describes the presence of inulin and oligofructose in common foodstuffs. Historical data on human consumption add an extra dimension. Modern analytical techniques (HPLC, LGC, HPAEC‐PAD) are used to check the variety of data mentioned in the literature throughout the past century. Methods to determine inulin and oligofructose in natural foodstuffs (cereals, fruit, and vegetables) are optimized and used to determine the loss of inulin during storage and during preparation of the food. These findings allow quantification of the amount of inulin and oligofructose in the average daily western diet. The daily per capita intake is estimated to range from 1 to 10 g, depending on geographic, demographic, and other related parameters (age, sex, season, etc.). Inulin and oligofructose are not measured by classic methods of dietary fiber analysis and consequently are often not mentioned in food tables. Their significant contribution (1 to 10 g/d/per capita) to the dietary fiber fraction (recommended at 25 g/d/per capita) is not taken into account in any nutritional recommendations. In view of this, inulin and oligofructose deserve more attention, both in food composition tables and in diet or nutrition studies.
With the increasing amount of evidence linking certain disorders of the human body to a disturbed gut microbiota, there is a growing interest for compounds that positively influence its composition … With the increasing amount of evidence linking certain disorders of the human body to a disturbed gut microbiota, there is a growing interest for compounds that positively influence its composition and activity through diet. Besides the consumption of probiotics to stimulate favorable bacterial communities in the human gastrointestinal tract, prebiotics such as inulin-type fructans (ITF) and arabinoxylan-oligosaccharides (AXOS) can be consumed to increase the number of bifidobacteria in the colon. Several functions have been attributed to bifidobacteria, encompassing degradation of non-digestible carbohydrates, protection against pathogens, production of vitamin B, antioxidants, and conjugated linoleic acids, and stimulation of the immune system. During life, the numbers of bifidobacteria decrease from up to 90 % of the total colon microbiota in vaginally delivered breast-fed infants to < 5 % in the colon of adults and they decrease even more in that of elderly as well as in patients with certain disorders such as antibiotic-associated diarrhea, inflammatory bowel disease, irritable bowel syndrome, obesity, allergies, and regressive autism. It has been suggested that the bifidogenic effects of ITF and AXOS are the result of strain-specific yet complementary carbohydrate degradation mechanisms within cooperating bifidobacterial consortia. Except for a bifidogenic effect, ITF and AXOS also have shown to cause a butyrogenic effect in the human colon, i.e., an enhancement of colon butyrate production. Butyrate is an essential metabolite in the human colon, as it is the preferred energy source for the colon epithelial cells, contributes to the maintenance of the gut barrier functions, and has immunomodulatory and anti-inflammatory properties. It has been shown that the butyrogenic effects of ITF and AXOS are the result of cross-feeding interactions between bifidobacteria and butyrate-producing colon bacteria, such as Faecalibacterium prausnitzii (clostridial cluster IV) and Anaerostipes, Eubacterium and Roseburia species (clostridial cluster XIVa). These kinds of interactions possibly favor the co-existence of bifidobacterial strains with other bifidobacteria and with butyrate-producing colon bacteria in the human colon.
The human gastrointestinal tract is colonised by a complex ecosystem of microorganisms. Intestinal bacteria are not only commensal, but they also undergo a synbiotic co-evolution along with their host. Beneficial … The human gastrointestinal tract is colonised by a complex ecosystem of microorganisms. Intestinal bacteria are not only commensal, but they also undergo a synbiotic co-evolution along with their host. Beneficial intestinal bacteria have numerous and important functions, e.g., they produce various nutrients for their host, prevent infections caused by intestinal pathogens, and modulate a normal immunological response. Therefore, modification of the intestinal microbiota in order to achieve, restore, and maintain favourable balance in the ecosystem, and the activity of microorganisms present in the gastrointestinal tract is necessary for the improved health condition of the host. The introduction of probiotics, prebiotics, or synbiotics into human diet is favourable for the intestinal microbiota. They may be consumed in the form of raw vegetables and fruit, fermented pickles, or dairy products. Another source may be pharmaceutical formulas and functional food. This paper provides a review of available information and summarises the current knowledge on the effects of probiotics, prebiotics, and synbiotics on human health. The mechanism of beneficial action of those substances is discussed, and verified study results proving their efficacy in human nutrition are presented.
Prebiotics are a group of nutrients that are degraded by gut microbiota. Their relationship with human overall health has been an area of increasing interest in recent years. They can … Prebiotics are a group of nutrients that are degraded by gut microbiota. Their relationship with human overall health has been an area of increasing interest in recent years. They can feed the intestinal microbiota, and their degradation products are short-chain fatty acids that are released into blood circulation, consequently, affecting not only the gastrointestinal tracts but also other distant organs. Fructo-oligosaccharides and galacto-oligosaccharides are the two important groups of prebiotics with beneficial effects on human health. Since low quantities of fructo-oligosaccharides and galacto-oligosaccharides naturally exist in foods, scientists are attempting to produce prebiotics on an industrial scale. Considering the health benefits of prebiotics and their safety, as well as their production and storage advantages compared to probiotics, they seem to be fascinating candidates for promoting human health condition as a replacement or in association with probiotics. This review discusses different aspects of prebiotics, including their crucial role in human well-being.
The development of functional food products is highly relevant due to declining public health and deteriorating food quality. Functional foods must not only meet the body's nutritional needs but also … The development of functional food products is highly relevant due to declining public health and deteriorating food quality. Functional foods must not only meet the body's nutritional needs but also provide therapeutic and preventive benefits. The aim of this study is to develop a functional fermented dairy product enriched with Jerusalem artichoke powder, honey, and pine nuts and to evaluate its nutritional value, probiotic activity, microbiological stability, and impact on blood sugar levels. The scientific significance of this study lies in proving the beneficial properties of Jerusalem artichoke in fermented dairy products. The practical value of this research is the development of an innovative product that can enhance health and nutrition quality, with the potential for industrial production. The methodology includes analyzing the chemical composition of ingredients, developing formulations, and studying the physicochemical, microbiological, and organoleptic properties of the final product. Its ability to lower blood sugar levels, strengthen immunity, and improve gut microbiota was assessed. As a result of the research, a technology for producing a functional fermented dairy product with Jerusalem artichoke powder was developed. The authors determined that it is low-calorie, highly biologically valuable, and possesses probiotic properties, making it a promising option for therapeutic and dietary nutrition.
Inulin and agavin fructans have been widely used in the food industry as fat substitutes, wall materials, and prebiotics, among other applications. Chemical modifications offer several advantages, from enhancing functional … Inulin and agavin fructans have been widely used in the food industry as fat substitutes, wall materials, and prebiotics, among other applications. Chemical modifications offer several advantages, from enhancing functional properties to broadening industrial applications, making them a key area of research in biotechnology, nutrition, and food science. This review examines the chemical modifications of fructans, specifically the inulin and agavin types. It describes the most commonly used methods, their characteristics, and their impact on the physicochemical, functional, and prebiotic properties of fructans. Additionally, it explores the interactions underlying these changes. Modifications enhance, extend, or generate new biological properties and activities. While most yield positive outcomes, challenges remain, including a deeper understanding of the structure–bioactivity relationships and further toxicity assessments, particularly in agavins. These insights aim to guide future research and innovation in the field.
Abstract Background Fructan content and flux in temperate forages can benefit the grasses through increased cold hardiness, increased drought tolerance, and improved forage quality. Orchardgrass ( Dactylis glomerata L., or … Abstract Background Fructan content and flux in temperate forages can benefit the grasses through increased cold hardiness, increased drought tolerance, and improved forage quality. Orchardgrass ( Dactylis glomerata L., or cocksfoot) produces relatively long and unbranched levan‐type fructans, but the genes involved in their biosynthesis are uncharacterized. Methods Through the evaluation of five orchardgrass cultivars and breeding lines that differ in their cold hardiness and freezing tolerance, we tested fructan and monosaccharide accumulation upon cold acclimation. The glycoside hydrolase‐32 ( GH32 ) gene family members involved in fructan biosynthesis were identified and grouped with homologous genes from Triticum aestivum and Lolium perenne . Results In each of four GH32 gene families, there were specific genes with high transcript levels and no deletions in GH32 motifs. The candidate for sucrose:fructan 6‐fructosyltransferase ( 6‐SFT ) exhibited the highest transcript levels of any GH32 gene in this study and was induced upon cold acclimation. Conversely, three invertase and two fructan exohydrolase genes, with roles in sucrose and fructan hydrolysis, had reduced transcript levels upon cold acclimation. Conclusions These data provide putative roles of GH32 genes in orchardgrass, and show that 6‐SFT , vacuolar invertase ( VI ), and fructan exohydrolases ( FEH ) genes play a role in fructan biosynthesis and metabolism for cold acclimation in orchardgrass.
This study aimed to produce invertase with characteristics comparable to commercial formulations using a low-cost raw material, employing environmentally friendly extraction and refinement methods. Sifted yeast, the residual baker's yeast … This study aimed to produce invertase with characteristics comparable to commercial formulations using a low-cost raw material, employing environmentally friendly extraction and refinement methods. Sifted yeast, the residual baker's yeast in industrial production, was used as raw material, and invertase in the yeast cell was extracted by ultrasonication and purified by micro- and ultra-filtration (MF and UF) methods. Subjecting the crude enzyme extract to MF followed by UF resulted in increased activity and specific activity. Through this process, the enzyme activity increased from 153 IU/mL to 691 IU/mL. The purified enzyme was lyophilized and the production of invertase at the laboratory scale was accomplished. The obtained enzyme was found to be stable at pH 5 and within the temperature range of 30-40 °C. It retained its activity for 60 days at -18 °C, whereas a 20% loss in activity was observed at +4 °C over the same period. As a result, it was demonstrated that invertase enzyme can be produced from a low-cost raw material which is considered as waste in the industry. To pass to the commercial production stage, processing of higher amounts of raw material by preventing foaming and heating problems in ultrasonication and scale-up studies testing the permeability properties of different membrane systems at a pilot-scale are necessary.
<title>Abstract</title> Levansucrase Ba-SacB from <italic>Bacillus amyloliquefaciens</italic> catalyses the production of levan using sucrose as substrate, and levan is widely used in food industry and medicine. Here the support of nano … <title>Abstract</title> Levansucrase Ba-SacB from <italic>Bacillus amyloliquefaciens</italic> catalyses the production of levan using sucrose as substrate, and levan is widely used in food industry and medicine. Here the support of nano zinc oxide is used for efficient immobilization of Ba-SacB(i.e., Ba-SacB-ZnO). FT-IR analysis revealed that Ba-SacB got absorbed via electrostatic interaction with the support. The Ba-SacB-ZnO showed improved activity in the pH range 3-5, and improved pH stability in the pH range 6-9. The Ba-SacB-ZnO kept nearly 70% activity after 21 d of storage, which was 20% higher than that of its free counterpart. In addition, the Ba-SacB-ZnO showed better tolerance to multiple chemicals, metal ions, organic solvents, proteases than the free Ba-SacB, suggesting that Ba-SacB-ZnO could be efficient for applications requiring in more diverse, especially harsh environments. About 50% of Ba-SacB-ZnO retained activity after 10 cycles of reuse. This study provides a theoretical basis for large-scale industrial applications of immobilized enzymes.
Abstract The growing demand for the sustainable and cost-effective production of probiotics and postbiotics has highlighted the potential of saline and marine plants as novel substrates. These plants, including seaweeds … Abstract The growing demand for the sustainable and cost-effective production of probiotics and postbiotics has highlighted the potential of saline and marine plants as novel substrates. These plants, including seaweeds and halophytes, are abundant and nutrient-rich and require minimal resources, making them ideal candidates for green biorefineries. The incorporation of saline plant-based feedstocks could lower media costs and environmental impact, as these plants do not require arable land or freshwater while contributing to carbon sequestration and sustainable farming. The development of integrated biorefineries could drive economic feasibility by facilitating cost-effective probiotic and postbiotic production. However, challenges such as high salt content and lignocellulosic composition may complicate microbial fermentation. This review examines recent advancements in leveraging naturally salt-tolerant probiotics and efficient bioconversion methods to address these challenges. It explores the nutritional profiles of saline plants, their prebiotic potential, and their synergetic compatibility with diverse probiotic strains, including probiotic bacteria and fungi and their metabolites. Additionally, the review discusses state-of-the-art fermentation techniques tailored to saline plant-based substrates and the possible advantages of saline feedstocks for probiotics and postbiotics production through biorefinery pathways. The work highlights the transformative potential of saline and marine plant-derived probiotics and postbiotics in health supplementation and biotechnological innovation, contributing to biorefinery development within a circular economy framework.
Leuzea cartamoides or Rhaponthicum cartamoides (hereinafter referred to as "Leuzea") is a valuable plant - a source of an ecdysterone. The aim of the research was to develop the technology … Leuzea cartamoides or Rhaponthicum cartamoides (hereinafter referred to as "Leuzea") is a valuable plant - a source of an ecdysterone. The aim of the research was to develop the technology of the syrup on the basis of the rhizomes and the roots of Rhaponthicum cartamoides, to study its composition and biological effect. The following objects were used as research objects: the rhizomes and the roots of Leuzea, from which a thick extract was obtained, auxiliary substances for the syrup preparation (a sucrose, a sorbitol, a glycerol, a citric acid, spinach green colouring agent, preservatives a nipagin and a nipazol, a propyleneglycol). A thick extract was previously prepared from the raw material of Leuzea. The syrup technology consisted of dissolving 40.0 а sorbitol in 40 ml of the purified water at 70-80°C. Then we dissolved in the sorbitol solution thick extract of the leuzeas the raw material 0.99; а citric acid 0.1; solution 1% from the spinach green colouring agent 2.0 ml; а nipagin 0.16 and а nipazole 0.02 in 2.0 propyleneglycol; а glycerol 5.0, and added а purified water to 100 ml. The syrup was a liquid of the green-brown colour, sour-sweet taste with the odour characteristic of the leuzeas preparations, with content of the tannins - 0.307±0.014%, of the acids free organic - 0.203±0.009%, the micronutrients essential (Na, K, Ca, P, Mg, Fe, Co, Cu, Ni, Mo, Zn, Mn) and the micronutrients conditionally essential (B, V, Si, Cr,). Biological studies have shown that the main component of the syrup - the extract from the leuzeas the raw material is not toxic, has adatogenic, antioxidant, membrane-stabilising effect.
Inulosucrase (IS) is a key enzyme in the synthesis of inulin, a multifunctional polysaccharide with significant physiological benefits and wide-ranging applications. Lactobacillus IS has the unique capability to produce both … Inulosucrase (IS) is a key enzyme in the synthesis of inulin, a multifunctional polysaccharide with significant physiological benefits and wide-ranging applications. Lactobacillus IS has the unique capability to produce both high-molecular-weight polysaccharides and oligosaccharides with diverse degrees of polymerization. Understanding the mechanism of sugar chain extension by IS is essential for modulating chain length and engineering custom-designed inulin. In this study, we resolved the crystal structures of IS from Lactobacillus reuteri 121 and its mutant IS-R544W, revealing a unique C-terminal extension into the catalytic pocket. Notably, structure-guided rational design identified IS-Tyr695 in the C-terminal region, along with IS-Asn303, IS-Asn305, IS-Asn367, IS-Gln369, and IS-Asn419, as critical residues specifically required for polysaccharide synthesis without affecting oligosaccharide production. In contrast, IS-Arg544, IS-Tyr618, and IS-Arg622 were determined to be essential for oligosaccharide synthesis with no impact on polysaccharide production. Based on findings from rational design and molecular dynamics simulations, we propose a novel shunting mechanism for the synthesis of polysaccharides and oligosaccharides by IS. This study provides fundamental insights into the inulin chain extension mechanism of IS and lays a theoretical foundation for engineering GH68 enzymes for the production of tailor-made fructans.
Raju K. Chalannavar , Ravindra B. Malabadi , Kiran P. Kolkar +5 more | Magna Scientia Advanced Biology and Pharmacy
Probiotics are defined as vital microorganisms which when ingested in suitable quantity offer a health benefit to the host. Probiotics are live microorganisms (Bifidobacterium, Lactobacillus, and a yeast, Saccharomyces boulardi … Probiotics are defined as vital microorganisms which when ingested in suitable quantity offer a health benefit to the host. Probiotics are live microorganisms (Bifidobacterium, Lactobacillus, and a yeast, Saccharomyces boulardi ) that can have beneficial effects on or inside body. Indian probiotic foods have a fascinating history deeply rooted in the country's rich cultural heritage. Ayurveda recognizes the importance of gut health and emphasizes the use of probiotic-rich foods to promote digestion, immunity, and overall well-being. Many probiotic products are formulated with beneficial bacteria (Bifidobacterium, Lactobacillus) and yeasts (Saccharomyces boulardii) for the purpose of preventing or recovering from bacterial or yeast infections in our different body parts, including, atopic dermatitis and acne, cavities and gum diseases, vaginal and urinary tract infections, and antibiotic associated diarrhea. It may also help to ease the symptoms of irritable bowel syndrome (IBS) and some other conditions. Indian probiotic food can alleviate digestive issues like bloating, gas, and constipation by promoting regular bowel movements. Indian probiotic foods may positively impact mental health by influencing gut microbiota composition and improving mood and cognitive function.
Abstract Inflammatory bowel disease is commonly treated with conventional therapies that suffer from poor drug bioavailability and significant side effects. This work develops a nanoparticle system using inulin, a naturally … Abstract Inflammatory bowel disease is commonly treated with conventional therapies that suffer from poor drug bioavailability and significant side effects. This work develops a nanoparticle system using inulin, a naturally occurring polysaccharide, to address these challenges. This colon‐targeted oral drug delivery system enables precise and efficient treatment of IBD by leveraging the colon‐specific degradation of inulin by the enzyme inulinase. The nanoparticles demonstrate high stability in the upper gastrointestinal tract and ensure the release of the drug payload in the colon, triggered by inulinase activity. In mouse models, oral administration of these nanoparticles, both with and without drug loading, significantly reduce body weight loss, preserve colon length, and exhibit synergistic anti‐inflammatory effects. Notably, even the nanoparticles without drug loading are effective in reducing inflammation and oxidative stress, likely due to the antioxidant properties of inulin. Additionally, the use of inulin promotes a beneficial shift in gut microbiota by decreasing pathogenic bacteria and increasing beneficial bacterial populations, further enhancing its therapeutic potential. These findings suggest that these prebiotic nanoparticles offer a targeted and effective strategy for IBD treatment by both anti‐inflammatory benefits and microbiota modulation. This study highlights the potential of prebiotic‐based nanomedicine for the treatment of IBD and possibly other colon‐related diseases.
The growing global need for circular bioeconomy processes has driven increasing interest in the valorization of fruit processing byproducts. Jaboticaba (Myrciaria jaboticaba) byproducts, such as bagasse, peel, and seeds, are … The growing global need for circular bioeconomy processes has driven increasing interest in the valorization of fruit processing byproducts. Jaboticaba (Myrciaria jaboticaba) byproducts, such as bagasse, peel, and seeds, are produced in large quantities by the juice industry and contain valuable technological and bioactive components, including dietary fibers and natural colorants (anthocyanins). While previous studies have primarily focused on their polyphenolic content, limited attention has been given to their dietary fiber and lignocellulosic composition. From this perspective, this study performed a comprehensive physicochemical, structural, and functional characterization of jaboticaba bagasse, peel, and seed flours, with a focus on their soluble and insoluble fibers, as well as their lignocellulosic components, including cellulose, lignin, and hemicellulose. Monosaccharide and disaccharide contents were quantified by HPAEC-PAD. Additionally, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), X-ray diffraction (XRD), scanning electron microscopy (SEM), anthocyanin quantification, and total phenolic content analysis were performed to assess the microstructure, technological properties, and bioactive potential. The results revealed high dietary fiber content and structurally diverse lignocellulosic matrices, suggesting potential applications, such as natural thickeners, stabilizers, and functional ingredients in food systems. This study expands the current understanding of jaboticaba byproducts and highlights their relevance for sustainable ingredient development, food innovation, and future applications in functional and clean-label formulations.
The high dependence on wheat flour and the increasing demand for functional foods for individuals with Diabetes Mellitus have spurred innovations in food products, including sourdough bread. This study aimed … The high dependence on wheat flour and the increasing demand for functional foods for individuals with Diabetes Mellitus have spurred innovations in food products, including sourdough bread. This study aimed to evaluate the effect of substituting 50% of wheat flour with varying ratios of purple sweet potato flour and brown rice flour on the sensory characteristics, proximate composition, and antioxidant activity of sourdough white bread. A Completely Randomized Design (CRD) was employed with four formulations: P0 (control/100% wheat flour), P1 (50:40:10), P2 (50:30:20), and P3 (50:20:30), each with three replications. Analyses included proximate composition (protein, fat, moisture, ash, carbohydrates), sensory attributes (color, aroma, taste, texture, overall acceptance), and antioxidant activity using the DPPH method. The results showed significant differences (p&lt;0,05) between the treatments and the control across all evaluated parameters. Based on the Zeleny test, the optimal formulation was P3, with protein content of 7,51 g, carbohydrate content of 48,86 g, and antioxidant activity (IC50) of 195,39 mg/mL. In conclusion, although the antioxidant activity was classified as weak, the bread formulation remains a promising candidate for functional food targeted at individuals with diabetes. It is recommended that 1,5 slices (approximately 50 g) of this sourdough bread be consumed as a snack for individuals with Diabetes Mellitus.
Este artículo analiza innovaciones en alimentos probióticos, destacando alternativas no lácteas que promueven la salud digestiva y el equilibrio de la microbiota intestinal. La microbiota cumple un rol fundamental en … Este artículo analiza innovaciones en alimentos probióticos, destacando alternativas no lácteas que promueven la salud digestiva y el equilibrio de la microbiota intestinal. La microbiota cumple un rol fundamental en funciones fisiológicas como la digestión, el metabolismo y la respuesta inmunitaria; mantener su balance mediante probióticos ayuda a prevenir enfermedades metabólicas, inflamatorias e, incluso, neuropsiquiátricas. Tradicionalmente, los alimentos probióticos eran productos lácteos como el yogur; sin embargo, la demanda por alternativas ha impulsado el desarrollo de alimentos no lácteos, incluyendo cereales, frutas, vegetales y snacks. Estos productos son diseñados para proporcionar los mismos beneficios que los lácteos y facilitar la inclusión de probióticos en la dieta sin alterar hábitos alimenticios. La selección correcta es fundamental para asegurar la efectividad de los probióticos, considerando aspectos como la especie y cepa bacteriana, la cantidad presente, y la lista completa de ingredientes. Sin embargo, en países como México, la falta de regulación clara en el etiquetado y seguridad de estos productos representa un desafío para el consumidor, quien debe basarse en etiquetado informativo y en recomendaciones expertas. La diversidad de estos alimentos permite una integración accesible en la dieta diaria y resalta su potencial como alternativas para mejorar la salud intestinal.
Tissue staining is necessary to show the structure and morphology of the tissue, namely Hematoxylin Eosin (H&amp;E) staining. However, eosin is carcinogenic when used in the long term. The aim … Tissue staining is necessary to show the structure and morphology of the tissue, namely Hematoxylin Eosin (H&amp;E) staining. However, eosin is carcinogenic when used in the long term. The aim of this research is expected to develop the use of natural materials as an alternative for Hematoxylin-Eosin histological staining. One of the color pigments that can be obtained from natural materials is the anthocyanin pigment found in strawberries. The anthocyanin extract from strawberries, which is acidic like eosin, will bind to positively charged molecules in the basic cytoplasm. The design of this study uses a simple experimental design (Posttest Only Design). Based on the Mann Whitney U test, a significant result was obtained with a value of sig.&lt;.001 &lt;0.05, indicating a significant difference in the results between the preparations stained with hematoxylin and eosin and those stained with hematoxylin and anthocyanin extract. Thus, it can be concluded that statistically, staining with hematoxylin and anthocyanin extract cannot be used as a substitute for hematoxylin eosin. Nevertheless, the results of staining the preparations using hematoxylin and anthocyanin extract still produce color in the cytoplasm, and are sufficiently contrasting to distinguish between the color of the cytoplasm and the color of the nucleus.