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Effect of carbon monoxide on fermentation of fiber, starch, and amino acids by mixed rumen microorganisms in vitro.

When 1 atm (101.3 kPa) of carbon monoxide was added to mixed rumen bacterial incubations containing timothy hay, methane production was inhibited by 88% without an increase in hydrogen. The molar ratio of propionate to acetate increased from 0.83 to 1.53, extracellular ammonia declined from 5.2 to 2.4 mM, and hemicellulose and cellulose digestions were inhibited by 40 and 27%, respectively. Even low levels of carbon monoxide (less than 0.1 atm [10.13 kPa]) significantly changed the products of fermentation. With starch, methane production was once again inhibited, but the magnitude of starch fermentation was unaffected. Decrease in acetate was accompanied by an equal molar increase in lactate. Ammonia production from the amino acid source, Trypticase, declined 20% as carbon monoxide was increased to 1.0 atm, and 93% of this decrease was explained by a selective inhibition of branched-chain amino acid fermentation.

Amino Acids↗

Postprocessing in vitro digestion challenge to evaluate survival of Escherichia coli O157:H7 in fermented dry sausages.

Fermented dry sausages, inoculated with Escherichia coli O157:H7 during batter preparation, were submitted to an in vitro digestion challenge to evaluate the extent to which passage through the human gastrointestinal tract could inactivate the pathogenic cells, previously stressed by the manufacturing process. The numbers of surviving E. coli O157:H7 cells remained constant after a 1-min exposure of the finely chopped sausage to synthetic saliva or during the following 120-min exposure to synthetic gastric juice at an initial pH of 2.0. However, significant (P < or = 0.05) growth of the pathogen (1.03 to 2.16 log10 CFU/g) was observed in a subsequent 250-min exposure to a synthetic pancreatic juice at pH 8.0. In a different set of experiments, fractions from the gastric suspension were transferred into the synthetic pancreatic juice at 30-min intervals to mimic the dynamics of gastric emptying. Concurrently, the pH of the remaining gastric fluid was reduced to 3.0, 2.5, and 2.0 to simulate the gradual reacidification of the stomach contents after the initial buffering effect resulting from meal ingestion. Under these new conditions, pathogen growth during pancreatic challenge was observed for the first few fractions released from the stomach (90 min of exposure [pH 2.5]), but growth was no longer possible in the fractions submitted to the most severe gastric challenge (120 min of exposure [pH < 2.2]).

Animals↗

Butyrate production from oligofructose fermentation by the human faecal flora: what is the contribution of extracellular acetate and lactate?

Butyrate is an important substrate for maintenance of colonic health and oligofructose fermentation by human faecal bacteria can increase butyrate production in vitro. However, oligofructose appears to be fermented by mainly acetate and lactate-producing bacteria rather than butyrate-producing bacteria. Isotope labelling studies using [U-(13)C(6)]glucose were used to show that (13)C(2) and (13)C(4) were the major labelled butyrate species produced from glucose fermentation, via [(13)C(2)]acetate-acetyl CoA as intermediate. Bacterial interconversion reactions were quantified and acetate conversion to butyrate and lactate conversion to acetate, propionate and butyrate were observed. Addition of oligofructose to faecal batch cultures significantly increased butyrate production. Of the newly synthesised butyrate from oligofructose fermentation, 80 % was derived from interconversion of extracellular acetate and lactate, with acetate being quantitatively more significant. Carbohydrates, such as oligofructose, have prebiotic properties. In addition, oligofructose selectively stimulates the bacterial conversion of acetate and lactate to butyrate. Carbohydrates with similar properties represent a refinement of the prebiotic definition, termed butyrogenic prebiotics, because of their additional functionality.

Acetates↗

In vitro activity of newer broad spectrum beta-lactam antibiotics against enterobacteriaceae and non-fermenters: a report from Austrian intensive care units. Austrian Carbapenem Susceptibility Surveillance Group.

We compared the in vitro activity of broad spectrum beta-lactam antibiotics against 573 gram-negative isolates (enterobacteriaceae and non-fermenters) collected between November 1996 and May 1997 from 9 laboratories serving intensive care units throughout Austria. MIC's (Minimal Inhibitory Concentration) were obtained with the E-test for meropenem, imipenem, ceftazidime, cefepime, cefpirome and piperacillin/tazobactam. Pseudomonas aeruginosa was the most frequently isolated organism (22%), followed by E. coli (19%), Klebsiella spp. (16%), and Enterobacter spp. (14%). Acinetobacter spp., Proteus spp., Serratia spp., Stenotrophomonas maltophilia, Citrobacter spp., Morganella morganii, Burkholderia cepacia and Salmonella enteritidis were isolated less frequently. Overall meropenem, imipenem and ceftazidime were the most active compounds in vitro, inhibiting 90%, 89%, and 87% of the isolates, respectively. Pseudomonas aeruginosa was inhibited by piperacillin/tazobactam in 89%, by cefepime in 87% and by ceftazidime in 85%. Imipenem, meropenem and cefpirome were less active (79%, 75% and 69% respectively). All E. coli strains were inhibited by meropenem, 99% were inhibited by imipenem, cefepime and cefpirome. Ceftazidime was active against 95% and piperacillin/tazobactam against 92% of E. coli. All Klebsiella spp. were inhibited by meropenem, cefepime and cefpirome. Imipenem inhibited 99% and ceftazidime 98% of the Klebsiella isolates. Piperacillin/tazobactam was active against 95% of Klebsiella spp. In vitro carbapenems are still the most active of all antibiotics tested. The relatively high resistance of Pseudomonas spp. and Acinetobacter spp. to carbapenems reflects the wide use of carbapenems during the last years. However, most bacterial isolates are still sensitive to the tested broad spectrum beta-lactams.

Acinetobacter↗

An in vivo 1H and 31P NMR investigation of the effect of nitrate on hypoxic metabolism in maize roots.

The effect of nitrate on the short-term hypoxic response and recovery of flooded mature maize roots has been investigated in vivo by 1H and 31P NMR and in vitro by 1H NMR and gas chromatography-mass spectrometry. Employing 1H NMR in addition to 31P NMR extended the number of identifiable compounds in vivo from 4 to 15, while in vitro two-dimensional NMR and gas chromatography-mass spectrometry aided rigorous in vivo 1H NMR resonance assignments and quantitation of 24 compounds. In the absence of nitrate, the concentrations of key metabolites including alanine, ethanol, gamma-aminobutyrate, lactate, succinate, and sucrose changed during 8 h of hypoxia in a manner consistent with reduced tricarboxylic acid cycle activity and diversion to glycolytic fermentation. The pH drop in the cytoplasm during hypoxia was rapid, about 0.2 unit, and diminished quickly upon recovery. Rapid recovery of ethanol, succinate, and sucrose levels was also observed, which indicates a return to normal aerobic metabolism. Although the hypoxic response itself, including pH, was not greatly affected by the presence of nitrate, nitrate reduced the amount of fermentation end products produced, helped maintain a higher free NTP concentration during hypoxia, and increased the rate of overall recovery from hypoxia. These findings suggest the presence of a nitrate-induced maintenance-level respiration in hypoxic maize roots, which helps explain the protection imparted by nitrate to flooded hypoxic maize plants.

Energy Transfer↗

Application of a tissue culture microtiter test for the detection of cytotoxic agents from natural products.

A method is described by which the growth inhibitory effects of cytotoxic compounds and fermentation broth cultures on adherent tumor cell lines can be quantitated. Cells are seeded into 96-well microtiter plates and 16 hours later the test compounds or broths are added to the wells. Cell growth is measured after three days (B16 mouse melanoma cells) or six days (HT-29, human colon carcinoma cells) by first fixing adherent cells, staining with Giemsa stain, washing away excess stain, then solubilizing stained cells with HCl. Absorbance is determined using a microELISA spectrophotometer and the data are transferred to and analyzed by a computer. The assay is rapid and reproducible and can be used to identify fermentation broths with cytotoxic components. Addition of DNA into the assay mixture (cells plus compound) inhibits the cytotoxic activities of certain DNA-reactive agents. The results of this study demonstrate the application of this assay system for primary and secondary evaluation of fermentation broths for in vitro antitumor activity.

Animals↗

Effect of cellulose fine structure on kinetics of its digestion by mixed ruminal microorganisms in vitro.

The digestion kinetics of a variety of pure celluloses were examined by using an in vitro assay employing mixed ruminal microflora and a modified detergent extraction procedure to recover residual cellulose. Digestion of all of the celluloses was described by a discontinuous first-order rate equation to yield digestion rate constants and discrete lag times. These kinetic parameters were compared with the relative crystallinity indices and estimated accessible surface areas of the celluloses. For type I celluloses having similar crystallinities and simple nonaggregating particle morphologies, the fermentation rate constants displayed a strong positive correlation (r2 = 0.978) with gross specific surface area; lag time exhibited a weaker, negative correlation (r2 = 0.930) with gross specific surface area. Crystallinity was shown to have a relatively minor effect on the digestion rate and lag time. Swelling of microcrystalline cellulose with 72 to 77% phosphoric acid yielded substrates which were fermented slightly more rapidly than the original material. However, treatment with higher concentrations of phosphoric acid resulted in a more slowly fermented substrate, despite a decrease in crystallinity and an increase in pore volume. This reduced fermentation rate was apparently due to the partial conversion of the cellulose from the type I to the type II allomorph, since mercerized (type II) cellulose was also fermented more slowly, and only after a much longer lag period. The results are consistent with earlier evidence for the cell-associated nature of cellulolytic enzymes of ruminal bacteria and suggest that ruminal microflora do not rapidly adapt to utilization of celluloses with altered unit cell structures.

Animals↗

A novel bone resorption inhibitor, A-75943 isolated from Streptomyces sp. SANK 61296.

A novel bone resorption inhibitor, A-75943, was isolated from Streptomyces sp. SANK 61296. Its structure was determined to be (1"S,2'R,3"S,4S',5"S)-2-[(3",5"- dimethyl-2"-oxocyclohexan-1"-yl)-6'-oxotetrahydropyran-4'-yl ]acetamide by spectral analyses and chemical conversion of cycloheximide. A-75943 inhibited bone resorption in vitro in a concentration-dependent manner with an IC50 of 0.35 microM, and also displayed an inhibitory effect on bone resorption in thyroid and parathyroid-extracted rats.

Animals↗

Use of HT-29, a cultured human colon cancer cell line, to study the effect of fermented milks on colon cancer cell growth and differentiation.

Epidemiological and in vivo and in vitro experimental studies have suggested that fermented milks may interfere with the emergence and/or the development of colon cancer. The results, however, remain inconclusive. This prompted us to develop a new approach based on the use of HT-29, a cultured human colon cancer cell line, to study at the cellular level the effect of fermented milks on colon cancer cell growth and differentiation characteristics. Undifferentiated HT-29 cells have been grown in the continuous presence of milks fermented by one of the following bacterial populations: Lactobacillus helveticus, Bifidobacterium, L.acidophilus or a mix of Streptococcus thermophilus and L. bulgaricus. Penicillin G was added to the cell culture medium, resulting in a complete blockade of bacterial growth without significant effect on bacterial viability. One out of the four bacteria species studied, namely L.acidophilus, was without effect on both cell growth and differentiation. The three other bacterial strains induced a significant, although variable, reduction in the growth rate of HT-29 cells, which resulted in a 10-50% decrease in the cell number at steady-state (i.e. at cell confluency). The most efficient strains in lowering the HT-29 growth rate were L. helveticus and Bifidobacterium. Concomitantly, the specific activities of dipeptidyl peptidase IV (DPP IV), a sensitive and specific marker of HT-29 cell differentiation, and that of three other brush border enzymes (sucrase, aminopeptidase N and alkaline phosphatase) were significantly increased, thus suggesting that these cells may have entered a differentiation process. Altogether, these results indicate that the use of cultured colon cancer cells may be a useful tool to further study the effect of fermented milks on colon cancer and that bacterial strains may exert a different and specific effect on cancer cell growth and differentiation when used in fermented milk products.

Animals↗

Application of the colon-simulation technique for studying the effects of Saccharomyces boulardii on basic parameters of porcine cecal microbial metabolism disturbed by clindamycin.

AIMS: The present study analyzed the effects of Saccharomyces boulardii on the biochemical parameters of microbial hindgut metabolism disturbed by clindamycin. METHODS: The experiments were carried out under in vitro conditions using the semicontinuous colon-simulation technique. This technique is standardized for quantitatively measuring parameters of microbial hindgut metabolism. The fluid and particle phase of pig hindgut contents were used for the in vitro incubations. The 5-day control period was followed by clindamycin exposure alone (312.5 mg/day for 5 days) or by a combined treatment of clindamycin and S. boulardii (400 mg/day for 5 days). RESULTS: Clindamycin resulted in significant decreases in production rates of short-chain fatty acids (SCFAs) which were associated with substantial changes in molar SCFA proportions at the expense of butyrate. These effects could at least partly be compensated for by S. boulardii, in particular by enhancements of acetate and propionate fermentation to control levels. In contrast, butyrate fermentation could not be reconstituted. In a second experiment the potential use of S. boulardii as a substrate for hindgut microbial metabolism was studied by comparing living and autoclaved yeast. Propionate and butyrate fermentation rates were unaffected whereas acetate fermentation tended to be higher in the presence of living yeast. CONCLUSIONS: S. boulardii can be effective to compensate for changes in microbial fermentation in response to antibiotic treatment. Despite the lack of statistical significance it might be concluded that the increase in fermentation end products can only partly be explained by the utilization of the yeast as a substrate for microbial metabolism.

Analysis of Variance↗

In Vitro Stimulation of Forage Fiber Degradation by Ruminal Microorganisms with Aspergillus oryzae Fermentation Extract.

Aspergillus oryzae fermentation extract (Amaferm) was evaluated for its ability to influence degradation of brome grass and switchgrass fiber fractions by mixed ruminal microorganisms in vitro. Addition of Amaferm at a concentration of 0.067 mg/ml, which is approximately the concentration found in the rumen ecosystem (0.06 mg/ml), increased the degradation of brome grass neutral detergent fiber (NDF) by 28% after fermentation for 12 h (P < 0.01), but had no effect after fermentation for 24 or 48 h. The levels of degradation of both the cellulose and hemicellulose fractions were increased after fermentation for 12 h (P < 0.01). Additions of 0.08 and 8% (vol/vol) Amaferm filtrate (12.5 g/100 ml) stimulated degradation of switchgrass NDF by 12 and 24% (P < 0.01), respectively, after fermentation for 12 h; when 80% filtrate was added, degradation was decreased by 38%. The concentrations of total anaerobes in culture tubes containing 80% filtrate were 5 times greater than the concentrations in the controls; however, the concentrations of cellulolytic organisms were 3.5 times lower than the concentrations in the controls (P < 0.05). These results suggested that the filtrate contained high concentrations of soluble substrate which did not allow the cellulolytic organisms to compete well with other populations. The remaining concentrations of esterified p-coumaric and ferulic acids were lower at 12 h in NDF residues obtained from fermentation mixtures supplemented with Amaferm. Because the total anaerobes were not inhibited in fermentation mixtures containing Amaferm, antibiotics are unlikely to be involved as a mode of action for increasing NDF degradation. The possibility that Amaferm contains enzymes (possibly esterases) that may play a role in stimulating the rate of fiber degradation by mixed ruminal microorganisms by removal of plant cell wall phenolic acid esters is discussed.

Journal Article↗

Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates.

A first aim of this batch in vitro experiment (21 h) was to use changes in odd and branched chain fatty acid (OBCFA) patterns to suggest shifts in microbial populations, associated with four types of incubated whole dairy cow diets. Principal component analysis suggested higher dietary starch increased the proportion of C15:0 and C17:0, whereas increased neutral detergent fibre content was positively related to anteiso C15:0 concentrations, which is in agreement with the importance of these fatty acids in respectively amylolytic and cellulolytic bacteria. A second aim of the experiment was to relate rumen volatile fatty acid proportions to OBCFA by principal component regression and to compare these relations with predictions based on diet proximate composition. The R2 values achieved for the regressions between acetate, propionate and butyrate, and OBCFA were 79.6%, 86.6% and 84.9% respectively. Moreover, in the current study, predictions of the rumen fermentation pattern showed higher R2 (p < 0.01) when based on OBCFA compared with proximate feed composition. If relations persist in vivo, there could be scope for milk OBCFA to predict the supply of specific rumen nutrients.

Animal Feed↗

Metabolism of mixed human colonic bacteria in a continuous culture mimicking the human cecal contents.

The purpose of this study was to develop an in vitro model of the proximal colon that would permit investigation of the function of colonic bacteria. Five continuous cultures of fecal bacteria were maintained anaerobically in steady state for at least 21 days in a medium simulating ileostomy effluent. The pH was maintained at 6 by the controlled infusion of sodium bicarbonate. Fresh medium (13 ml) was pumped into the culture for 2 min every hour and excess culture was simultaneously removed. The redox potential of all cultures remained below -300 mV, whereas the osmolality was maintained hypertonic compared with the original inoculum or medium. The cultures readily produced volatile fatty acids at a rate of 30.4 +/- 3.6 mmol/day (mean +/- SEM). Acetic acid (10-19 mmol/day) and propionic acid (5-30 mmol/day) were produced in the largest amounts, whereas butyric acid was produced only in small amounts or not at all. Ammonia was produced in each culture in amounts that varied from day to day and between cultures. The osmolality and volatile fatty acid production increased when the carbohydrate input was doubled and decreased when the cultures were deprived of carbohydrate. Thus it is possible to maintain actively fermenting viable cultures in vitro for prolonged periods; such cultures respond to changing conditions and may be useful in investigating the metabolic function of the colon.

Bacteria↗

Effect of Escherichia coli wild type or its derivative with high nitrite reductase activity on in vitro ruminal methanogenesis and nitrate/nitrite reduction.

The effects of two kinds of Escherichia coli strains, wild-type E. coli W3110 or E. coli nir-Ptac, which has enhanced nitrite reduction activity, on in vitro CH4 production and nitrate and nitrite reduction in cultures of mixed ruminal microorganisms was investigated using continuous incubation systems. Escherichia coli nir-Ptac, a derivative of wild-type E. coli W3110, was constructed by replacing self promoter of nir BD operon encoding subunits of nitrite reductase in E. coli W3110 by tac promoter to make the expression of nir BD higher and constitutive. The nitrite reductase activity of E. coli nir-Ptac was approximately twice as high as E. coli W3110. The culture media consisted of 400 mL of strained ruminal fluid taken from two nonlactating Holstein cows receiving a basal diet of orchardgrass hay at maintenance level (55 g of DM/kg of BW0.75 daily), and 400 mL of autoclaved artificial saliva. Treatments were arranged in two separate 3 x 3 factorials consisting of nitrate (NaNO3; 0, 5, or 10 mM) without E. coli or inoculated with E. coli W3110 or E. coli nir-Ptac, or nitrite (NaNO2; 0, 1 or 2 mM) without E. coli or inoculated with E. coli W3110 or E. coli nir-Ptac. The control culture contained no chemical or microbial additives. Escherichia coli cells were inoculated into in vitro mixed ruminal cultures at approximately 2 x 10(8) to 10(9) cells/mL. Methane production by ruminal microorganisms was decreased markedly (P < 0.001) by the addition of nitrate and nitrite, and by the inoculation of cultures with E. coli W3110 or E. coli nir-Ptac (P < 0.01). With mixed nitrite-containing cultures, E. coli nir-Ptac inhibited (P < 0.001) in vitro nitrite accumulation and CH4 production more than E. coli W3110, which may be due to the tac promoter-enhanced nitrite reductase activity of E. coli nir-Ptac accelerating electrons to be consumed for nitrite reduction rather than CH4 biosynthesis. In conclusion, anaerobic cultures of E. coli W3110 or E. coli nir-Ptac may decrease CH4 production in the rumen. The inoculation of E. coli W3110 or, especially, E. coli nir-Ptac to mixed ruminal microorganisms may decrease nitrite toxicity when ruminants consume high-nitrate-containing forages and when nitrite is applied to abate ruminal CH4 production.

Animals↗

Dietary influence on caecal microbial nitroreduction of a drug: metronidazole.

Six diets were fed to weanling rats and the in vitro caecal microbial nitroreduction rates of metronidazole were determined. The sensitivity of this metabolic pathway to diet was demonstrated by the observed 20-fold range in reaction rate constant. Methods were devised to investigate the mechanism of this dietary effect. It was concluded that indigestible dietary components affected primarily both the weight of caecal contents and the metabolic activities of the microorganisms per unit weight of caecal contents. It appeared that the actual reaction rate in the caecum might be 35% higher than that determined by the standard assay procedure that involves sample dilution. However, the different soluble materials in the caeca derived from rats fed different diets had no significant secondary regulatory effects on the rate constant.

Anaerobiosis↗

Relationship between the in vitro-estimated utilizable crude protein and the Cornell Net Carbohydrate and Protein System crude protein fractions in feeds for ruminants.

The objectives of this experiment were to study the regression relationship between the in vitro-estimated utilizable crude protein (uCP) and the Cornell Net Carbohydrate and Protein System (CNCPS) crude protein fractions, i.e. PA, PB1, PB2, PB3 and PC of feeds for ruminants, and the possibility of using CNCPS crude protein fractions for the estimation of uCP of feeds for ruminants. The CP, the non-protein nitrogen (NPN), the soluble cru de protein (SCP), the neutral detergent insoluble crude protein (NDICP) and the acid detergent insoluble crude protein (ADICP) of 30 single feeds were determined and the CP fractions, i.e. PA, PB1, PB2, PB3 and PC, were then calculated based on CNCPS. The uCP of the 30 feeds was estimated using the in vitro incubation technique of Zhao and Lebzien (2000). It was found that there was a significant multiple regression relationship between the uCP [g/kg dry matter (DM)] and the CNCPS crude protein fractions, i.e. PA (% DM), PB1 (% DM), PB2 (% DM), PB3 (% DM) and PC (% DM) of the feed samples. uCP = (9.95 +/-2.73)PA + (2.92 +/- 1.36)PB(1) + (7.24 +/- 0.86)PB(2) + (8.20 +/- 3.33)PB(3) + (17.67 +/- 3.79) PC + (63.26 +/- 18.02), r2 = 0.90, n = 30, p < 0.0001. It was suggested that the multiple regression relationship shown above could be used for the estimation of the uCP of feeds based on the CNCPS crude protein fractions PA, PB1, PB2, PB3 and PC of feeds for ruminants.

Animal Feed↗

Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria.

New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains.

Bacteria↗

Antihypertensive effect of peptides obtained from Enterococcus faecalis-fermented milk in rats.

Previous studies have demonstrated that milk fermented with Enterococcus faecalis decreases the systolic blood pressure (SBP) and the diastolic blood pressure (DBP) of spontaneously hypertensive rats. In this study, we evaluated the antihypertensive activity of the following peptide sequences: LHLPLP, LHLPLPL, LVYPFPGPIPNSLPQNIPP, VLGPVRGPFP, and VRGPFPIIV. These peptides isolated from E. faecalis-fermented milk showed in vitro angiotensin I-converting enzyme-inhibitory activity. Because the most potent angiotensin I-converting enzyme-inhibitory sequences were LHLPLP and LVYPFPGPIPNSLPQ-NIPP, we administered different doses of these peptides to spontaneously hypertensive rats. High doses of the remaining sequences were also administered to these animals. Water served as a negative control and captopril as a positive control. All products were administered orally. The SBP and DBP were measured before administration and also at 2, 4, 6, 8, and 24 h after administration. Before administration of the different products, spontaneously hypertensive rats showed SBP and DBP values of 218 +/- 2.5 and 157 +/- 5.9 mmHg, respectively (n = 30). The sequences LHLPLP, LVYPF-PGPIPNSLPQNIPP, VLGPVRGPFP, and VRGPFPIIV caused clear and significant decreases in SBP, DBP, or both in the animals. In particular, the antihypertensive effect could be clearly established when 2 or 3 mg/kg of LHLPLP was administered. These 2 doses of LHLPLP showed similar antihypertensive properties. Four hours after administration of captopril or the highest doses of the different peptides, the decreases in the SBP and the DBP (mmHg) were as follows: captopril (SBP = 52 +/- 5.8, DBP = 38.8 +/- 3.8), 3 mg/kg of LHLPLP (SBP = 25.3 +/- 8.2, DBP = 29.5 +/- 7.6), 6 mg/kg of LVYPFPGPIP-NSLPQNIPP (SBP = 14.9 +/- 3.7, DBP = 8.7 +/- 4.4), 10 mg/kg of LHLPLPL (SBP = 7.7 +/- 4.1, DBP = 9.4 +/- 3.1), 10 mg/kg of VLGPVRGPFP (SBP = 16.2 +/- 5.8, DBP = 21.64 +/- 3.2), and 10 mg/kg of VRGPFPIIV (SBP = 16.05 +/- 2.74, DBP = 9.19 +/- 3.49). The results obtained suggest that the sequences LHLPLP, LVYPFPGPIPNSLPQ-NIPP, VLGPVRGPFP, and VRGPFPIIV could be responsible, at least in part, for the antihypertensive properties described for E. faecalis-fermented milk.

Animals↗