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PubMedDate not supplied

Evaluation of the cariogenic potential of cassava flours from the Amazonian region.

Cassava flour is the main dietary carbohydrate source in the Amazonian region. The cariogenic potentials of the two main kinds of cassava flour - 'seca' and 'd'água' - were evaluated in vivo and in vitro. A sweet made from a regional fruit (cupuaçú) and a species of local fish (tambaqui) were used as positive and negative controls, respectively. For in vivo evaluation of dental plaque acidogenicity, the study had a crossover design, in four legs, with 19 volunteers in four treatment groups. Dental plaque pH was determined by the microtouch method before and for 60 min after food consumption. For in vitro evaluation, stimulated saliva of each volunteer was incubated with the food and pH variation was determined over 4 h. Carbohydrates were chemically determined in the flours and the majority was found to be water-insoluble. In vivo, plaque pH decreased significantly after the consumption of the sweet (p < 0.05), increased after the fish (p < 0.05), but did not change after intake of the flours (p > 0.05). In vitro, the flours were slowly fermented by bacteria present in saliva. The in vivo and in vitro findings suggest that, in the form that the main cassava flours from the Amazonian region of Brazil are customarily eaten, they may be considered to have no or very low cariogenicity.

Adult↗
PubMed2006

The isoflavonoid aglycone-rich fractions of Chungkookjang, fermented unsalted soybeans, enhance insulin signaling and peroxisome proliferator-activated receptor-gamma activity in vitro.

We investigated anti-diabetic candidates and their mechanisms from the fractions of Chungkookjang (CKJ), a traditional fermented unsalted soybean, by investigating insulin signaling, peroxisome proliferator-activated receptor (PPAR)-gamma activity and glucose-stimulated insulin secretion, in vitro. Cooked soybeans (CSB) and CKJ, fermented predominantly with Bacillus subtilis, were extracted by 70% EtOH followed by an XAD-4 column chromatography with a serial mixture of solvents comprised of MeOH and water. During fermentation, the contents of isoflavonoid aglycones were elevated, and the fractions enriched with aglycones enhanced insulin-stimulated glucose uptake in 3T3-L1 adipocytes. This increase in glucose uptake resulted from stimulating a translocation of the glucose transporter (GLUT)-4 into the plasma membrane through the phosphorylation of insulin receptor substrate (IRS)-1 and Akt. Especially, daidzein enriched fractions elevated insulin-stimulated glucose uptake by acting as PPAR-gamma agonist up to levels exhibited when 10 nM insulin is administered. Fractions containing small peptides with low polarity in CKJ slightly increased glucose-stimulated insulin secretion. The data suggest that an increase in isoflavonoid aglycones in CKJ, in comparison to CSB, enhances glucose utilization via activating insulin signaling and stimulates PPAR-gamma activity in adipocytes. In addition, CKJ contains small peptides improving glucose-stimulated insulin secretion in insulinoma cells. Overall, CKJ is superior to CSB in anti-diabetic action.

3T3-L1 Cells↗
PubMed2003

Improvement of the digestibility of the proteins of the red alga Palmaria palmata by physical processes and fermentation.

Palmaria palmata (dulse) is an edible red alga constituting a potential protein source in human diet. However, previous studies showed that the digestibility of dulse proteins is bad because of the cell-wall encapsulating cytoplasmic proteins and the presence of fibers. The water-soluble xylan, present in high proportions in dulse, could be involved to explain the weak digestibility of proteins. To limit the influence of fibers and to improve the nutritional quality of these proteins, we have treated dulse by physical processes or by fermentation by moulds. After a 30 min predigestion by pepsin followed by a 6 h digestion into a cell dialysis containing porcine pancreatin, the corrected in vitro digestibility of crude dulse was very low (about 1.5% after correction by digestibility blank). The in vitro protein digestibility was estimated to 58% of that of casein for dulse samples obtained after washing in demineralized water and grinding in liquid nitrogen. The in vitro protein digestibility of fermented samples was 45%-65% of that of casein. After physical treatment, the digestibility improvement was related to the elimination of soluble molecules such as xylan and mineral salts. The improvement observed after fermentations seemed due to the degradation of insoluble fibers.

Aspergillus↗
PubMed1983

Survival rate of Salmonella and Shigella in fermented milk products with and without added human gastric juice: an in vitro study.

The survival rates of Salmonella agona, Salmonella java, Salmonella typhimurium and Shigella sonnei in milk and fermented milk products were investigated with and without the addition of human gastric juice during a 7 to 10 hour test period. It was found that yoghurt inhibited the growth of Salmonella and Shigella very effectively even when the yoghurt had been heated to 100 degrees C for 15 minutes, whereas milk and the other fermented milk products showed a lower ability to inhibit the growth of pathogens. Yoghurt plus human gastric juice greatly depressed the growth rate of the pathogens; after 30 minutes no more colonies were formed. Even kefir or ropy milk plus gastric juice showed inhibition of Salmonella typhimurium after one hour. In the case of acidophilus milk plus gastric juice, the inhibition of Salmonella typhimurium occurred first after 2.5 hours and the presence of viable Shigella sonnei was noticed for 4 hours. The addition of physiological NaCl instead of human gastric juice to the fermented milk samples before inoculation changed the picture of the survival rate. Only yoghurt inhibited the growth of Salmonella java after 30 minutes; Shigella sonnei survived in yoghurt for 4 hours and in the other fermented milk samples for 5 hours. It can be assumed that yoghurt contains some antimicrobial compounds that inhibit the growth of pathogens and that this inhibiting property is enhanced by the addition of human gastric juice.

Animals↗
PubMed2005

Shiga toxin production by sausage-borne Escherichia coli O157:H7 in response to a post-processing in vitro digestion challenge.

In order to study the effects of the fermentation-drying procedure and subsequent in vitro digestion on Shiga toxins (Stx) production by Escherichia coli O157:H7, dry sausages were inoculated during the formulation step with pure cultures of strains 5-1 and ATCC 43895. The inoculated sausages were submitted to a minimum (30 min, pH between 3.1 and 3.5) or a maximum (120 min at stepwise adjusting the pH downward) gastric challenge followed by a 240-min pancreatic challenge at pH 8.0 and 37 degrees C. Production of toxins by the overnight culture controls, assessed using the Vero cell assay, was dependent on the pathogen cell concentration. The effect of cell concentration was not relevant in sausage samples and data showed: (a) higher Stx production in contaminated sausage samples than in overnight cultures; (b) the lowest Stx levels were detected with undigested sausage samples; (c) the maximum gastric challenge enhanced Stx production, compared to minimally digested and undigested samples. Reverse transcriptase polymerase chain reaction (RT-PCR) performed on extracts from inoculated, digested (4.5-6 h process) and undigested sausages produced amplicons for both stx1 and stx2 mRNA, suggesting that post-stress expression of stx genes had occurred. Our data suggest that sub-lethal stresses imposed by the fermentation-drying procedure and subsequent digestion of ingested food (i.e. contaminated sausages) may affect the degree to which the surviving E. coli O157:H7 cells express their virulence in vivo.

Animals↗
PubMed1978

Some effects of arsenic on the rumen microflora; an in vitro study.

The rate of fermentation of the rumen microflora was inhibited almost 30% by 5 micrograms/ml of arsenic added in the form of arsenite, although 304 micrograms/ml was required to cause 50% inhibition. Arsenate was less inhibitory. The rate of fermentation of a separated bacterial fraction was inhibited 37% by 1 microgram of arsenite per millilitre, whereas 100 micrograms/ml had little effect on the fermentation of a separated protozoal fraction. Similar results were obtained for arsenate. Both fractions had the capacity to take up arsenate, but the protozoa took it up more readily to a higher intracellular concentration. Both arsenate and arsenite inhibited the growth of a number of rumen bacteria in pure culture at concentrations as low as 5 micrograms of arsenic per millilitre. The greater resistance of Megasphaera elsdenii to arsenate as compared with that of Bacteroides succinogenes was not related to the inability to take up the element. In conclusion, the concentrations of arsenic causing a significant inhibitory effect on the fermentative activity and growth of some rumen bacteria are less than that reported to be toxic to ruminant animals.

Anaerobiosis↗
PubMed1995

Degradation and fermentation of alpha-gluco-oligosaccharides by bacterial strains from human colon: in vitro and in vivo studies in gnotobiotic rats.

The ability of several human gut bacteria to break down alpha-1,2 and alpha-1,6 glycosidic linkages in alpha-gluco-oligosaccharides (GOS) was investigated in vitro in substrate utilization tests. Bacteroides thetaiotaomicron, Bifidobacterium breve and Clostridium butyricum, which are usually found in the infant gut and have been associated with both beneficial and deleterious effects on health, were studied. Alpha-Gluco-oligosaccharide degradation was compared in vitro and in vivo in gnotobiotic rats associated with these organisms, inoculated alone or in combination. Oligomer breakdown and short chain fatty acid and gas production indicated hydrolysis and fermentation of the substrate. In vitro and in vivo, Cl. butyricum was the least efficient in utilizing GOS, whereas Bact. thetaiotaomicron was the most efficient. Kinetic studies on GOS hydrolysis in pH-regulated fermenters showed that alpha-1,2 glucosidic bonds, which characterize the substrate, were more resistant than alpha-1,6 linkages. Adaptation of gnotobiotic rats to a diet containing 2% (w/w) GOS significantly increased the hydrolysis of alpha-1,2 glucosidic bonds. Combination of bacteria in trixenic rats improved GOS degradation and inhibited Cl. butyricum metabolism. This inhibition was confirmed in pH-regulated fermenters containing GOS as the principal carbon source. The association of beneficial bacteria and GOS may therefore have a potential health-promoting effect in human neonates.

Animals↗
PubMed2006

In vitro anti-cancer activity of a novel microbial fermentation product on human carcinomas.

The possible anti-proliferation and cell death induction potential of a novel microbial fermentation extract named as oncogen XP-180 (or simply as XP-180) was tested on three human solid tumour carcinoma cell lines (non-small cell lung cancer A549, breast cancer MDA-MB231, liver adenocarcinoma SK-Hep1) and on the acute myelogenous leukaemia KG1a cell line. Anti-proliferative activity of XP-180 was observed on all of these cancer cell lines with comparable efficiency and in a dose-dependent manner. Morphological investigation further suggested that common features of apoptosis, including cell shrinkage and rounding, are present in XP-180 treated cells. Loss of adhesion properties of these solid tumour cell lines was observed upon XP-180 incubation. Anchorage-dependent clonogenicity assay on solid tumour cell lines and semi-solid methylcellulose colony formation assay on leukaemia cell line further revealed that XP-180 strongly inhibited the regeneration potential of these cancer cells. Using KG1a as an experimental model system, XP-180 was shown to stimulate the activity of caspase 3, 8 and 9 without significant change in caspase 6 activity. Furthermore, XP-180 readily induced collapse of mitochondrial membrane potential after 2 h of incubation. However, the use of the generic caspase specific inhibitor Z-VAD-FMK does not significantly reverse XP-180 mediated cell death. The results obtained suggest that XP-180-mediated cancer cell death could involve mitochondria and both caspase-dependent and -independent pathways. Therefore, XP-180 is an efficient anti-cancer regimen in vitro.

Amino Acid Chloromethyl Ketones↗
PubMed2003

Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro.

A series of in vitro studies was conducted to determine the effects of adding a commercial enzyme product on the hydrolysis and fermentation of cellulose, xylan, and a mixture (1:1 wt/wt) of both. The enzyme product (Liquicell 2500, Specialty Enzymes and Biochemicals, Fresno, CA) was derived from Trichoderma reesei and contained mainly xylanase and cellulase activities. Addition of enzyme (0.5, 2.55 and 5.1 microL/g of DM) in the absence of ruminal fluid increased (P < 0.001) the release of reducing sugars from xylan and the mixture after 20 h of incubation at 20 degrees C. Incubations with ruminal fluid showed that enzyme (0.5 and 2.55 microL/g of DM) increased (P < 0.05) the initial (up to 6 h) xylanase, endoglucanase, and beta-D-glucosidase activities in the liquid fraction by an average of 85%. Xylanase and endoglucanase activities in the solid fraction also were increased (P < 0.05) by enzyme addition, indicating an increase in fibrolytic activity due to ruminal microbes. Gas production over 96 h of incubation was determined using a gas pressure measurement technique. Incremental levels of enzyme increased (P < 0.05) the rate of gas production of all substrates, suggesting that fermentation of cellulose and xylan was enzyme-limited. However, adding the enzyme at levels higher than 2.55 microL/g of DM failed to further increase the rate of gas production, indicating that the maximal level of stimulation was already achieved at lower enzyme concentrations. It was concluded that enzymes enhanced the fermentation of cellulose and xylan by a combination of pre- and postincubation effects (i.e., an increase in the release of reducing sugars during the pretreatment phase and an increase in the hydrolytic activity of the liquid and solid fractions of the ruminal fluid), which was reflected in a higher rate of fermentation.

Animal Feed↗
PubMedDate not supplied

Effect of anaerobic fungi on in vitro feed digestion by mixed rumen microflora of buffalo.

Five strains of anaerobic fungi isolated from the faeces of wild (hog deer, Cervus porcinus; blackbuck, Antelope cervicapra; spotted deer, Axis axis; nilgai, Baselophus tragocamelus) and rumen liquor of domestic (sheep, Ovies aries) ruminants showing high fibrolytic enzyme producing ability were added to mixed rumen microflora of buffalo to study their effect on the digestibility of lignocellulosic feed (wheat straw and wheat bran in the ratio of 80:20), enzyme production and fermentation end products in in vitro conditions. Among the 5 isolates studied, FNG5 (isolated from nilgai) showed the highest stimulating effect on apparent digestibility (35.31 +/- 1.61% vs. 28.61 +/- 1.55%; P < 0.05), true digestibility (43.64 +/- 1.73% vs. 35.37 +/- 1.65%; P < 0.01), neutral detergent fiber digestibility (29.30 +/- 2.58% vs. 18.47 +/- 2.12; P < 0.01) of feed 24 h after inoculation compared to the control group. The production of carboxymethyl cellulase, xylanase, acetyl esterase and beta-glucosidase was significantly (P < 0.05) higher in the FNG5 inoculated incubation medium. There was no improvement in the digestibility and enzyme production on the addition of the other 4 isolates. Total volatile fatty acid levels as well as the concentration of acetate, propionate, isobutyrate and valerate were significantly higher in the FNG5 added group as compared to the control group. The fungal isolate FNG5 from nilgai, a wild ruminant, was found to be superior to the other isolates tested and appears to have a potential to be used as a feed additive for improving fiber degradation in domestic ruminants.

Animal Feed↗
PubMed1994

[Effects of orally administered therapeutic drugs on the fermentation processes in the rumen fluid of ruminating cattle (in vitro). 6. Copper sulfate].

In 50 trials, incubating rumen fluid at 39 degrees C for 6 hours with starch and urea as nutrients, the effects of simulated oral administration of 3 g copper sulphate per animal and day (50 mg copper sulphate per liter rumen fluid) on rumen fermentation patterns have been examined. In two series of five incubations each in rumen fluid of five donors (total = 50), doses of 2.25 g (37.5 mg/l) and 4.5 g (75 mg/l) copper sulphate were simulated. The five donors (517-720 kg) of ruminal fluid were fed a ration of hay and concentrate twice daily and water was offered ad libitum. Rumen fluid was sampled 2.5 hours after feeding in the morning and immediately incubated in the artificial rumen. While at 2.25 g an effect was hardly seen and 4.5 g were followed by great changes, 3 g appeared to be the limiting dose that reduced the amylolysis (-23.9%; p < 0.001), the production of volatile fatty acids (-23.5%; p < 0.001) and methane (-23.6%; p < 0.001), but caused a higher concentration of ammonia (+11.2%; p < 0.001). Therefore 2 g CuSO4/animal/day is recommended as maximum dose in cattle to overcome Cu-deficiency.

Administration, Oral↗
PubMed2000

Effect of sausage ingredients and additives on the production of enterocin A and B by Enterococcus faecium CTC492. Optimization of in vitro production and anti-listerial effect in dry fermented sausages.

Enterocin A and B in Enterococcus faecium CTC492 were co-induced by the different factors assayed in this study (r = 0.93) and followed primary metabolic kinetics. Enterocin production was significantly inhibited by sausage ingredients and additives, with the exception of nitrate. The addition of sodium chloride and pepper decreased production 16-fold. The temperature and pH influenced enterocin production, with optima between 25 and 35 degrees C, and from 6.0 to 7.5 of initial pH. The maximum activity was achieved, under favourable growth conditions, with MRS supplemented with sucrose (2%) plus glucose (0.25%) and Tween-80 (1%). MRS concentration, NaCl plus pepper addition, absence of Tween-80 in the growth medium, incubation at 45 degrees C and an initial pH under 5.5 were detrimental to bacteriocin production. Stress conditions did not favour enterocin production. Desadsorption was Tween-dependent. Enterocin A activity in the crude extracts stored at -80 degrees C was better preserved than enterocin B (when tested against their specific indicator strain), but anti-listerial activity remained intact. Applied as anti-listerial additives in dry fermented sausages, enterocins significantly diminished Listeria counts by 1. 13 log (P < 0.001), while Enterococcus faecium CTC492 added as starter culture did not significantly reduce Listeria counts (P > 0. 1) compared with the standard starter culture (Bac-). Enterocins A and B could be considered as extra biopreservative hurdles for listeria prevention in dry fermented sausages.

Bacteriocins↗
PubMed1992

The colon in carbohydrate malabsorption: short-chain fatty acids, pH, and osmotic diarrhoea.

Short-chain (C2-C6) fatty acids (SCFA) are the major anions in colonic contents and the result of anaerobic fermentation of mainly saccharides. The effects and regulation of saccharide fermentation were studied in vitro and in vivo. In vitro faecal incubation was used to study the effects of lactose, glucose, and galactose and of pH on SCFA formation. Changing the pH to below 5 or above 11 abolished SCFA formation in the faecal incubates; in the pH 5-9 interval SCFA production was high, with only minor pH dependence. Adding glucose, galactose, or lactose to the incubation system increased SCFA production, but at high saccharide concentrations (100-300 mmol/l) SCFA formation was inhibited by the pH change. In vivo disaccharide malabsorption with increasing doses of lactulose caused a decrease in faecal pH to less than 5, values inhibitory to fermentation, before the appearance of carbohydrate in faeces. In 6 of 12 volunteers diarrhoea occurred suddenly and was caused by malabsorbed non-fermented carbohydrate. The six other volunteers had a gradual increase in faecal output with lactulose dose and developed diarrhoea before the appearance of saccharide in faeces. The intake of lactulose tolerated before diarrhoea ensued varied between individuals, with the majority having diarrhoea of more than 11/day at 160 g lactulose per day. At this dose SCFA absorption was estimated to be in the range 550 to 1150 mmol/day.

Colon↗
PubMed1999

In-vitro activity of cefepime and seven other antimicrobial agents against 1518 non-fermentative Gram-negative bacilli collected from 48 Canadian health care facilities. Canadian Afermenter Study Group.

Non-fermentative bacilli are primarily nosocomial pathogens, and are also often resistant in vitro to a broad range of antimicrobial agents. In this large Canadian study, we collected 1466 clinical, non-repeat isolates of Pseudomonas aeruginosa, 21 of Acinetobacter spp. and 31 Stenotrophomas maltophilia. MICs of eight antibiotics were determined by the NCCLS microdilution method in a central laboratory. Tobramycin was the most active agent against P. aeruginosa (94.5% susceptible); amikacin and imipenem were the most active against Acetinobacter spp. (100%) and ceftazidime was the most active against S. maltophilia (40.6%). Against each group of isolates, cefepime was active against 87, 86.4 and 15.6%, respectively. This in-vitro study showed that cefepime may be a useful additional agent in the treatment of infections caused by P. aeruginosa and Acinetobacter spp., but not when S. maltophilia is considered pathogenic.

Anti-Bacterial Agents↗
PubMed2001

Development of a simple in vitro assay for estimating net rumen acid load from diet ingredients.

The objective of these experiments was to develop a simple in vitro technique for evaluating the production and neutralization of acid as feeds ferment in the rumen. An in vitro approach was adopted to eliminate animal factors. The procedure was based on the method of Tilley and Terry, with some modifications developed in this project. Residual acidity (acidogenicity value) was determined by the dissolution of Ca from CaCO3 powder added to the media at the end of 24-h incubations. Acidogenicity values (AV) were higher when 20% strength buffer was used, while lowering buffer pH increased values, equally across all feeds. There was no effect of donor animal diet, but considerable day-to-day variation in the fermentation activity of rumen fluid. This variation likely reflected the substrate preferences of differing microbial populations, so that several standard feeds may be required to account for this effect. A series of 28 diverse feed ingredients was evaluated for AV using a mixture design, with 85 combinations of ingredients: 100% of each ingredient (n = 28); 50% of each ingredient and an equal mixture of all others (n = 28); equal mixture of all ingredients, excluding one (n = 28); and an equal mixture of all ingredients (n = 1). The effects of most ingredients on AV were essentially linear, though some extreme ingredients showed nonlinear effects. Protein sources had low AV, forages intermediate AV and starchy feeds high AV. Calcium contained within feeds contributed to AV, particularly for legumes, sugar beet pulp, and citrus pulp, and must be accounted for.

Animal Feed↗
PubMed2026

Simmondsia chinensis (jojoba) cake fermentation: A new, sustainable technology for advanced skin and scalp care ingredients.

OBJECTIVE: Simmondsia chinensis is a well-known commercially popular plant from which jojoba oil is extracted. Jojoba cake is a sustainably produced, intractable by-product of the jojoba seed oil extraction currently used principally as a fertilizer or burned as fuel. Fermentation work conducted with various microorganisms, including Lactobacillus plantarum, Saccharomyces cerevisiae and Streptococcus thermophilus, sustainably grown on aqueous jojoba cake fermented the cake, liberating jojoba-based amino acids, peptides and proteins. The ferments have been examined chemically and via in&#xa0;vitro cell and tissue studies to develop new skin and scalp care targeted ingredients. METHODS: The jojoba cake contained nutrients (proteins, sugars and lipids) that self-sustain aqueous bacterial fermentation. The ferments were examined on 3D tissue models in&#xa0;vitro via human genomic microarrays. A ferment produced by Lactobacillus plantarum was further tested in&#xa0;vitro using ELISA protein assays on skin cell cultures. A 56-day clinical study on 46 individuals examined the influence of 1.0% of the Lactobacillus ferment on collagen expression using Diffuse Reflectance Spectroscopy (DRS). RESULTS: Gene responses were measured on 244+ genes known to have skin functions. It was found that the Lactobacillus ferment showed the greatest upregulation of skin-associated genes, and three highly upregulated proteins were examined more closely in&#xa0;vitro using ELISA protein assays: collagen-1A1, protocadherin-18 and opioid growth factor receptor. Each protein was upregulated in a dose-dependent fashion. The collagen analysis by DRS demonstrated a statistically significant increase in collagen fluorescence on Day 28 and Day 56 compared to baseline and placebo cream. Further mapping of the collagen fluorescence was done on the individuals using the active formulation at Days 0, 28 and 56. CONCLUSION: Jojoba cake presents a new source of sustainably grown biomass, but the cake is not suitable for topical applications. Fermentation produces components more suitable for topical care. In&#xa0;vitro studies demonstrated upregulation of three skin proteins associated with healing skin. Further studies also employed a newly emerging spectroscopic technique to measure collagen fluorescence in the skin in&#xa0;vivo, the results supporting in&#xa0;vitro work indicating the ferment made with Lactobacillus was able to stimulate collagen synthesis in the skin.

Lactobacillus↗