Effect of Lactic Acid Bacteria on the Pharmacokinetics and Metabolism of Ginsenosides in Mice

This study aims to investigate the effect of lactic acid bacteria (LAB) on in vitro and in vivo metabolism and the pharmacokinetics of ginsenosides in mice. When the in vitro fermentation test of RGE with LAB was carried out, protopanaxadiol (PPD) and protopanaxadiol (PPD), which are final metabolites of ginsenosides but not contained in RGE, were greatly increased. Compound K (CK), ginsenoside Rh1 (GRh1), and GRg3 also increased by about 30%. Other ginsenosides with a sugar number of more than 2 showed a gradual decrease by fermentation with LAB for 7 days, suggesting the involvement of LAB in the deglycosylation of ginsenosides. Incubation of single ginsenoside with LAB produced GRg3, CK, and PPD with the highest formation rate and GRd, GRh2, and GF with the lower rate among PPD-type ginsenosides. Among PPT-type ginsenosides, GRh1 and PPT had the highest formation rate. The amoxicillin pretreatment (20 mg/kg/day, twice a day for 3 days) resulted in a significant decrease in the fecal recovery of CK, PPD, and PPT through the blockade of deglycosylation of ginsenosides after single oral administrations of RGE (2 g/kg) in mice. The plasma concentrations of CK, PPD, and PPT were not detectable without change in GRb1, GRb2, and GRc in this group. LAB supplementation (1 billion CFU/2 g/kg/day for 1 week) after the amoxicillin treatment in mice restored the ginsenoside metabolism and the plasma concentrations of ginsenosides to the control level. In conclusion, the alterations in the gut microbiota environment could change the ginsenoside metabolism and plasma concentrations of ginsenosides. Therefore, the supplementation of LAB with oral administrations of RGE would help increase plasma concentrations of deglycosylated ginsenosides such as CK, PPD, and PPT.

Comparison of in situ techniques to evaluate the recovery of indigestible components and the accuracy of digestibility estimates

Indigestible components, including indigestible dry matter (iDM) and indigestible neutral detergent fiber (iNDF), play an integral role as internal markers for determining ruminal kinetics and digestibility estimations. However, the accuracy of internal markers is dependent upon the incubation technique utilized as bag type (BT) and incubation length (IL) can be significant sources of error. Previous studies have primarily focused on iDM and iNDF as digestibility markers, but few studies have compared digestibility estimates to those of acid detergent insoluble ash (ADIA). Therefore, our objective was to investigate the effect of BT (F57, F58, and Dacron) and IL (288 and 576 h) on iDM and iNDF residues, DM and NDF digestibilities, and fecal recoveries when using in situ incubations. Additionally, we evaluated the accuracy of digestibility estimates when using iDM, iNDF, and ADIA. For iDM and iNDF, feed residues demonstrated a BT × IL interaction (P < 0.01). However, fecal residues were only influenced by the main effects of BT and IL (P < 0.01), with the F58 BT and 288-h IL having the greatest residues for both iDM and iNDF. The variation in residues was greatly reduced when using iNDF compared with iDM. Fecal recovery estimates most closely approximated 100% recovery when utilizing ADIA and iDM using the F57 × 576 h incubation method (P < 0.01), although recovery was overestimated for all incubation combinations. Fecal NDF recovery estimates better represented the excretion profiles when the F57 × 576 h combination was used with iDM as the internal marker (P < 0.01). Estimates of DM and NDF digestibility were the most accurate when utilizing ADIA (P < 0.01) relative to all other treatments. Our results indicate that the proper methodological application is specific to the purpose of the inferences. When evaluating fecal recoveries and digestibility, ADIA or iDM with F57 at 576-h in situ incubation provides the greatest accuracy.

Use of n-alkanes to estimate feed intake in ruminants: a meta-analysis

Precise techniques to estimate feed intake by ruminants are critical to enhance feed efficiency and to reduce greenhouse gas emissions and nutrient losses to the environment. Using a meta-analysis, we evaluated the accuracy of the n-alkane technique to predict feed intake in cattle and sheep and assessed the relationships between feed intake and fecal recovery (FR) of n-alkanes. The database was composed of 28 studies, including 129 treatments (87 and 42 for cattle and sheep, respectively) and 402 animals (232 cattle and 170 sheep) fed at troughs, from published studies. Relationships between observed (in vivo measurement) and predicted feed intake by C31:C32 and C32:C33 n-alkane pairs were evaluated by regression. Meta-regression addressed the relationships between the difference in FR of n-alkane pairs and the error in intake estimation, as well as the amount and duration of C32 n-alkane dosing. Regression of observed intake on n-alkane-based estimates revealed good relationships in cattle (adjusted R2 = 0.99 for C31:C32, and adjusted R2 = 0.98 for C32:C33; P < 0.0001) and in sheep (adjusted R2 = 0.94 for C31:C32, and adjusted R2 = 0.96 for C32:C33; P < 0.0001). FR of natural n-alkanes showed a coefficient of variation of about 15% and 16% for C31 and C33, respectively, in cattle. In sheep, the coefficient of variation was 8% and 14% for C31 and C33, respectively. The relationships between the difference of FR of n-alkane pairs and the error in feed intake estimation in cattle were characterized by an adjusted R2 = 0.83 for C31:C32 (P < 0.0001) and adjusted R2 = 0.93 for C32:C33 (P < 0.0001). In sheep, they were characterized by an adjusted R2 = 0.69 for C31:C32 (P < 0.001) and adjusted R2 = 0.76 for C32:C33 (P < 0.001). The n-alkane technique provided the reliability for estimating feed intake in cattle and sheep in barn experiments. The present meta-analysis demonstrated that without correction for differences in FR of n-alkane pairs, deviation in feed intake prediction would occur. However, further research is necessary to determine the relationship between the n-alkane dosing procedure (daily amount and duration of dosing) and FR of n-alkane.

Fecal Recovery of Probiotics Administered as a Multi-Strain Formulation during Antibiotic Treatment

The present study aimed to investigate whether probiotic recovery is affected when consumed together with antibiotics. Fecal samples were collected from an earlier antibiotic associated diarrhea, randomized, placebo-controlled study with a product consisting of a combination of Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, and Bifidobacterium lactis Bi-07, B. lactis Bl-04 at equal numbers and at a total dose of 1010 CFU. Fecal samples were collected during the screening visit (T0), i.e., at the time of antibiotic prescription, and then on the last day of the antibiotic treatment (T1) as well as seven days after the subject had stopped taking the antibiotic treatment (T2) and at two weeks after completing antibiotic treatment and one week after probiotic/placebo consumption stopped (T3). Samples were analyzed for the presence of the four administered strains. The study was registered at clinicaltrials.gov as NCT01596829. Detection levels of all four strains were significantly increased from T0 to T1 and returned to baseline level from T2 to T3. There were also significantly more subjects with detectable levels of L. paracasei Lpc-37, B. lactis Bi-07, and B. lactis Bl-04 at T1 and T2 compared to T0 and T3, and compared to placebo. Each of the four strains could be detected in the feces of patients apparently unaffected by the simultaneous consumption of antibiotics.

The use of portable X-ray fluorescence spectrometry to measure apparent total tract digestibility in beef cattle and sheep

The use of portable X-ray fluorescence (PXRF) spectrometry to detect external markers on processed or unprocessed cattle and sheep fecal specimens to estimate apparent total tract digestibility (ATTD) was evaluated. Exp. 1: ruminally cannulated Angus-crossbred steers (n = 7; BW = 520 ± 30 kg) were individually fed ad libitum for 21 d in a completely randomized design (CRD). Markers (Cr2O3 and TiO2) were placed inside the rumen twice daily (7.5 g of each marker). Fecal samples were collected twice daily from day 14 to 21. Exp. 2: crossbred wethers (n = 8; BW = 68 ± 3 kg) were individually fed ad libitum for 21 d in a CRD. During this period, 2 g of Cr2O3 and TiO2 were top-dressed onto the feed twice daily. Sheep were housed in metabolism crates for 5 d for total fecal collection. Concentration of markers was determined on diets, refusals, and fecal specimens (fresh, dry-only, and dried/ground) using atomic absorption to detect Cr and spectrophotometry for Ti. Concentration of both markers was also determined via the PXRF spectrometer. Delta between ATTD estimated by wet chemistry and PXRF was not different from zero (P ≥ 0.14) when using cattle fresh fecal specimens for both markers, whereas ATTD estimated by PXRF with dry-only and dried/ground fecal specimens were 3.6 and 1.1 percent units lower (P ≤ 0.04), respectively, than ATTD estimated by wet chemistry for Cr and Ti, respectively. Regardless of the fecal sample preparation method on cattle specimens, Ti concentration was similar (P = 0.39) among methodologies, while Cr was underestimated (P < 0.01) by 13% when PXRF was used in dry-only or dried/ground samples. The ATTD of sheep was underestimated (P < 0.01) by 2.4 percent units compared with control when Cr was measured by PXRF in dry-only samples. The Cr concentration in dry-only fecal specimens of sheep tended (P = 0.09) to be lower compared with wet chemistry analysis. Fresh and dry/ground sheep fecal samples assessed for Cr, and dry-only assessed for Ti were not (P ≥ 0.49) affected by detection method. The Cr fecal recovery tended (P = 0.10) to be the lowest for dry-only, the greatest for wet chemistry, intermediate for fresh and dry/ground sheep-fecal specimens; while not affected (P = 0.40) for Ti. The PXRF is an accurate technology to detect Cr and Ti in fresh cattle fecal samples to estimate ATTD. For fresh and dry/ground, the technology was effective for determining the concentration of Cr, or dry-only fecal specimens when detecting Ti in sheep specimens.

Pharmacokinetics and Intestinal Metabolism of Compound K in Rats and Mice

We aimed to investigate the plasma concentration, tissue distribution, and elimination of compound K following the intravenous administration of compound K (2 mg/kg) in rats and mice. The plasma concentrations of compound K in mice were much higher (about five-fold) than those in rats. In both rats and mice, compound K was mainly distributed in the liver and underwent biliary excretion. There was 28.4% fecal recovery of compound K in mice and 13.8% in rats, whereas its renal recovery was less than 0.1% in both rats and mice. Relative quantification of compound K and its metabolite protopanaxadiol (PPD) in rat bile and intestinal feces indicated that the metabolism from compound K into PPD occurred in the intestine but not in the plasma. Therefore, PPD detected in the plasma samples could have been absorbed from the intestine after metabolism in control rats, while PPD could not be detected in the plasma samples from bile duct cannulated rats. In conclusion, mice and rats shared common features such as exclusive liver distribution, major excretion pathway via biliary route, and intestinal metabolism to PPD. However, there were significant differences between rats and mice in the plasma concentrations of compound K and the fecal recovery of compound K and PPD.

PSI-35 Comparison of in situ techniques to evaluate the recovery of indigestible components and the accuracy of digestibility estimates

Our objective was to evaluate the effects of bag type (BT) and incubation length (IL) when using in situ incubations for determination of indigestible components. Indigestible components play an integral role as an internal marker for rumen kinetics and digestibility estimations, as well as providing parameter estimates for nutritional models. However, the accuracy of internal markers is dependent upon the incubation technique utilized, with BT and IL acting as sources of error. A 3 × 2 factorial design was used to investigate the effect of BT (F57, F58, and Dracon) and IL (288 and 576 h) on indigestible dry matter (iDM) and neutral detergent fiber (iNDF) residues, as well as estimated DM and NDF digestibilities. Feed and fecal samples were obtained from two animals from an independent trial that collected total fecal production over 4 days. All bags contained a sample size-to-surface area ratio of 20 mg/cm2 and were ruminally incubated within three rumen-cannulated steers (229.5 ± 10 kg). For iDM and iNDF, feed residues demonstrated a BT × IL interaction (P < 0.01). Fecal residues for iDM and iNDF had BT and IL effects (P < 0.01) with the Dracon BT and 576 h IL having the least residues. Digestibility estimates for DM and NDF were influenced by BT × IL (P < 0.01) with the accuracy of estimates demonstrating a BT × IL interaction (P < 0.01) in which F57 at 576 h had the greatest accuracy. Resultant fecal recovery estimates for DM and NDF excreted exhibited a BT × IL interaction (P < 0.01), with F57 at 576 h better representing the excretion profiles. Based upon our analysis, utilizing the F57 BT at 576 h IL combination provides more accurate estimates relative to all other combinations used within the current study.

Titanium dioxide (TiO2) as a marker to estimate fecal output in sheep

ABSTRACT: Titanium dioxide (TiO2) is an external marker used to estimate fecal output based on forage intake of grazing ruminants; however, its use as marker for grazing sheep still needs attention. In the present study the TiO2 role to estimate fecal output of sheep grazing annual ryegrass was tested. Two essays were conducted in sequence using the same forage and animals. Six lacaune lambs were housed in metabolic cages. Lambs were administered 1 g TiO2 in a single (at 8:30 am, essay 1), or in two daily doses (at 8:30 am and 4:30 pm, essay 2). In both essays the TiO2 administration was followed by a 7-day period to stabilize the fecal excretion of the marker and then by a 5-day period for total collection of feces. Observed and TiO2-based estimated fecal outputs were compared, with or without correction for TiO2 fecal recovery rate (kTiO2). The kTiO2 was around 1.0 and the estimated fecal outputs were similar to the observed ones (p>0.05), regardless the number of daily doses. These results suggested that TiO2, administered in single or two daily doses, can be used as an external marker in grazing sheep.

n-Alkane and long-chain alcohol recovery in moose (Alces alces), a browsing herbivore

Habitat management for herbivores often depends on an understanding of the food habits of animals. Plant cuticular waxes containing nearly indigestible complex mixture of n-alkanes and long-chain alcohols (LCOHs) have recently shown promise for diet analyses, but the accuracy of the technique depends strongly on the efficiency of recovery of the markers in feces. Fecal recovery of n-alkanes and LCOHs from 10 browse stems or leaves and two ensiled grass hays fed to moose (Alces alces (Linnaeus, 1758)) during in vivo digestion trials was investigated. n-Alkanes and LCOHs were extracted using a single-step accelerated solvent extraction technique and the recovery of these cuticular components was calculated from the feces of the animals. n-Alkane recoveries from feces averaged 0.82, ranging from a low of 0.58 (haylage) to a high of 0.95 (browse stems). LCOH recoveries averaged 0.92 across all forages, ranging from 0.80 (haylage) to a high of 1.13 (browse stems). n-Alkane and LCOH fecal recovery increased with increasing chain length, similar to findings in other studies. Although fecal recovery of n-alkanes and LCOHs were variable, we conclude that they are inversely related to forage digestibility, are consistent within forage classes, and are therefore predictable markers for use in assessing herbivore diets.

Evaluation of internal markers as determinants of fecal dry matter output and digestibility in feedlot sheep

This study aimed to evaluate the fecal recovery of the internal markers indigestible dry matter (iDM), indigestible neutral detergent fiber (iNDF), and indigestible acid detergent fiber (iADF) and to compare them as estimators of the fecal dry matter output (FDMO) and digestibility of the dry matter (DM) with values observed in total feces collection in trials with feedlot sheep. For this, 82 sampling units from two experiments were evaluated. Internal markers were quantified in the feed supplied, orts, and feces. The recovery of the markers was determined by the comparison of the amount of marker consumed and excreted. Fecal dry matter output and digestibility were determined based on the amount of marker consumed and on its concentration in the feces through individual collections directly from the rectum of the animals, with values compared with those observed in the total feces collection from orthogonal contrasts. The mean bias, the concordance correlation coefficient, and the mean squared prediction error were also evaluated. For all statistical procedures, the critical level of probability for type-I error was fixed at 0.05. The marker iDM showed complete fecal recovery (100.03%) and was efficient in estimating FDMO (482.9 g day-1) and DM digestibility (542 g kg-1) as compared with the values observed in the total feces collection (454.7 g day-1 and 561 g kg-1 for FDMO and DM digestibility, respectively). The iNDF and iADF markers did not show complete recovery in the feces and were not effective in the estimate of FDMO and DM digestibility.