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PubMed2002

A high-performance liquid chromatography/tandem mass spectrometric screening method for eight synthetic corticosteroids in bovine feces and the simultaneous differentiation between dexamethasone and betamethasone.

A screening method was developed to monitor the illegal use of synthetic corticosteroids in cattle. Diethyl ether extracts from spiked feces samples were cleaned-up by solid phase extraction followed by semipreparative reversed-phase chromatography (RPC). The fraction containing the corticosteroids was derivatized with ethoxyamine hydrochloride. The corresponding ethoximes were separated using silica-based C18 RPC and analyzed on-line in an ion trap mass spectrometer using atmospheric pressure positive chemical ionization. Ethoxime derivatives of dexamethasone and betamethasone were baseline resolved, allowing for the simultaneous mass spectrometric differentiation of both epimers in bovine feces by conventional non-chiral chromatography. At the lowest level tested (1 micro g/kg), corticosteroids (except triamcinolone) could be identified in compliance with the recent European criteria for residue identification. The quantitative performance of the method was best at residue levels > or = 2 micro g/kg.

Animals↗
PubMed1995

Isotope dilution gas chromatographic-mass spectrometric method for the determination of unconjugated lignans and isoflavonoids in human feces, with preliminary results in omnivorous and vegetarian women.

We describe an isotope dilution gas chromatographic-mass spectrometric (GC/MS) method for the identification and quantitative determination of the lignans enterolactone, enterodiol, and matairesinol and the isoflavonoids daidzein, equol, O-desmethylangolensin, and genistein in feces. Following the addition of deuterated internal standards for all compounds, the feces samples are extracted and purified in several ion exchange chromatographic steps. Following formation of trimethylsilyl ethers, the samples are analyzed by combined capillary column GC/MS in the selective ion monitoring mode and corrected for all losses during the procedure using the deuterated internal standards. Results on the reliability of the method and values for nine Finnish omnivorous and nine vegetarian women are presented.

4-Butyrolactone↗
PubMed2002

Comparison of multiplex PCR, PCR-ELISA and fluorogenic 5' nuclease PCR assays for detection of plasmid-bearing virulent Yersinia enterocolitica in swine feces.

Swine are implicated as the principal animal reservoir for plasmid-bearing Yersinia enterocolitica (YEP(+)) strains that are pathogenic to humans. To evaluate the utility of the PCR for detection of YEP(+) strains in naturally-contaminated pig feces, samples were first enriched in Irgasan ticarcillin potassium chlorate broth for 48 h at 25 degrees C and then tested by multiplex PCR, PCR-ELISA, and fluorogenic 5' nuclease PCR assays. Three different primer sets for amplification of the ail gene sequences were used in these three assays. Three out of 50 (6%) samples were positive for YEP(+) strains using the multiplex PCR targeting the chromosomal ail (170 bp) and plasmid virF (591 bp) genes. Two of the 3 samples positive by the multiplex PCR were also positive by the PCR-ELISA method using primers targeting the ail gene (425 bp). In contrast, the fluorogenic 5' nuclease PCR assay failed to detect an ail gene sequence (118 bp) in any of the 50 samples. These results indicate that the multiplex PCR was the most reliable and sensitive assay for detecting YEP(+) strains in feces among the three assays evaluated.

Animals↗
PubMed1982

Enumeration of Methanobrevibacter smithii in human feces.

A plating medium containing cephalothin and clindamycin was developed for enumeration and isolation of methanogens in human feces. Specimens from nine CH4-producing subjects had total anaerobe counts of 1-8 x 1011/g dry weight. Methanogen counts on the antibiotic medium ranged from 0.001 - 12.6% of the total anaerobe count. There was no correlation between age, sex or percent dry fecal weight and the ratio of methanogens to total counts. Specimens from eight non-CH4-producing individuals contained bacteria that yielded nonmethanogenic colonies on the antibiotic medium. The means +/- SD of the logarithm of the total counts per gram dry weight were 11.4 /+- 0.29 and 11.38 +/- 0.44 for the positive and negative groups respectively. Values for the antibiotic-resistant flora were 8.8 /+- 1.13 and 7.78 /+- 1.08 respectively. Methanogens were isolated from the most dilute inoculum of each specimen from CH4-producing subjects. All isolates were morphologically, physiologically, and immunologically identical to Methanobrevibacter smithii. Growth of methanogens in media that were essentially extracts of CH4-negative feces suggested that no nutrients were lacking or inhibitors present in intestinal contents that prevent the growth of methanogens in these individuals.

Adult↗
PubMed1995

Isolation, purification, and biological activity of mono- and dihydroxylated paclitaxel metabolites from human feces.

Three metabolites of the cytotoxic drug paclitaxel (Taxol) were isolated and purified from the feces of cancer patients receiving the agent as an intravenous infusion. The procedures involved sample homogenization in water followed by liquid-liquid extraction with diethyl ether and high-performance liquid chromatography (HPLC). Approximately 1-3.5 mg of each metabolite was obtained from 100 g of feces. As judged from the chromatographic traces of analytical HPLC with ultraviolet (UV) detection at 227 nm, the purity of each compound was > 97%. On-line photodiode-array detection demonstrated that the UV spectrum of the isolated compounds closely resembles that of the parent drug. Mass spectrometry provided evidence that these metabolites are mono- and dihydroxy-substituted derivatives, namely, 6 alpha-hydroxypaclitaxel, 3'-p-hydroxypaclitaxel, and 6 alpha, 3'-p-dihydroxypaclitaxel. The two 6 alpha-hydroxy-substituted metabolites were shown to have lost their cytotoxicity in in vitro clonogenic assays using the A2780 human ovarian carcinoma and the CC531 rat colon-carcinoma tumor cell lines. In addition, the metabolites showed reduced myelotoxic effects as compared with paclitaxel in an in vitro hemopoietic progenitor toxicity assay. Our procedure for the isolation and purification of paclitaxel metabolites in milligram quantities should be useful for testing the biological activities of these compounds and for the preparation of calibration standards essential for pharmacokinetics studies.

Animals↗
PubMed1989

Glycosphingolipids in feces of germ-free rats as a source for studies of developmental changes of intestinal epithelial cell surface carbohydrates.

Non-acid and acid glycosphingolipids were isolated from feces of one litter of germ-free rats from day 17 to day 51. Quantitative and qualitative changes described for small intestine of conventional rats [Bouhours D, Bouhours J-F (1981) Biochem Biophys Res Commun 99:1384-89] were also found in the feces of these germ-free rats. A decrease in lactosylceramide and sialyllactosylceramide excretion and a change from N-acetylneuraminic acid to N-glycoloylneuraminic acid, as well as an appearance of type 1 chain blood group H-active penta- and decaglycosylceramides were observed during the weaning period. Thus the dramatic changes seen in rat intestinal glycosphingolipids postnatally seem to be primarily regulated by non-microbial factors.

ABO Blood-Group System↗
PubMed1980

Coronavirus-like particles in the feces of normal cats.

Coronavirus-like particles, morphologically indistinguishable from coronavirus-like particles seen in human, canine, and simian feces, were detected by electron microscopy in the feces from both feline infectious peritonitis antibody-positive and antibody-negative cats.

Animals↗
PubMedDate not supplied

Listeria isolations from feces of patients with diarrhea and from healthy food handlers.

A study was undertaken on the presence and frequency of Listeria sp. in feces from 1,000 patients suffering from diarrheal diseases and from 2,000 healthy persons. Furthermore, the feces of patients were examined for other well-documented enteropathogens such as Campylobacter, Salmonella, Shigella, Staphylococcus aureus, Yersinia enterocolitica, protozoa and rotavirus as well as for organisms of questionable enteropathogenic potency such as fungi, i.e. Candida. Finally, in continuation of previously described investigations of the enteropathogenic role of Proteus mirabilis but not of Proteus vulgaris, both these species were studied too. Only Listeria innocua and Listeria monocytogenes could be detected in the investigated fecal specimens. There were no differences of the frequencies of L. innocua, and L. monocytogenes between patients and healthy persons. 17 strains (= 1.7%) of L. innocua and six strains (= 0.6%) of L. monocytogenes were isolated from 1,000 samples of patients. As a comparison 2,000 fecal samples from healthy people contained 40 strains (= 2.0%) of L. innocua and 16 strains (= 0.8%) of L. monocytogenes. A coincidence study showed that there were no statistically significant correlations between well-known enteropathogens and Listeria sp., Proteus sp. or any of the other isolates. Significant correlations were found only between harmless species such as L. innocua and P. vulgaris.

Diarrhea↗
PubMed1986

Incidence and virulence of Aeromonas species in feces of children with diarrhea.

Aeromonas spp. occurring in feces of children with diarrhea were studied. Forty-eight strains were isolated from 2,025 specimens during a one year period. Only 11 of 44 strains tested yielded virulence factors (cytotoxin, hemolysin and hemagglutinin). Six strains were identified as Aeromonas sobria and five as Aeromonas hydrophila. The other strains isolated were identified as Aeromonas caviae. The biochemical characteristics associated with virulence factors were a positive Voges-Proskauer reaction, production of gas from glucose, fermentation of mannose, and absence of beta-lactosidase. Beta-D-glucosidase and esculin hydrolysis were the main characteristics used to differentiate Aeromonas sobria from the other two species. The incidence of Aeromonas spp. with virulence factors in feces of children with diarrhea would seem to vary widely from one area to another.

Adolescent↗
PubMed1996

A newly developed PCR assay of H. pylori in gastric biopsy, saliva, and feces. Evidence of high prevalence of H. pylori in saliva supports oral transmission.

We have recently developed a new PCR assay for the detection of H. pylori. In this study, the polymerase chain reaction (PCR) assay was used to detect H. pylori in 88 gastric biopsy, 85 saliva, and 71 fecal specimens from 88 patients. H. pylori infection was confirmed in 71 of 88 patients by culture and/or histological stain of gastric biopsies. Serum IgG antibody to H. pylori was also measured and resulted in 97% sensitivity and 94% specificity. H. pylori DNA was detected by the PCR assay in gastric biopsy specimens from all 71 patients (100% sensitivity) with proven gastric H. pylori infection but not from 17 noninfected patients (100% specificity). In saliva specimens, H. pylori DNA was identified in 57 of the 68 patients (84%) with proven gastric H. pylori infection and in three of the 17 patients without gastric H. pylori infection. However, the PCR assay was only able to detect H. pylori DNA in the feces from 15 of 61 patients (25%) with proven gastric H. pylori infection and one of the 10 patients without gastric H. pylori infection. The results show that the PCR assay is reliable for detecting the presence of H. pylori in gastric biopsy and saliva specimens. The data indicate that H. pylori exists in a higher prevalence in saliva than feces and that the fecal-oral route may be an important means of transmission of this infection in developing countries but not as significant as previously suspected in the developed countries. It is likely that the oral-oral route is more prominent.

Antibodies, Bacterial↗
PubMedDate not supplied

The metabolism and kinetics of 125I-labeled human intravenous IgG preparation (C-425) in rats and rabbits. I. Blood clearance, excretion into urine and feces, and brain uptake.

The metabolism of intravenous human IgG preparation (C-425) was studied by the use of 125I-C-425 in rats and rabbits after intravenous injection of the compound particularly with regard to changes in the levels of radioactive compound or metabolites in the blood as a function of time as well as levels excreted in urine and feces. Brain uptake index of 125I-C-425 was estimated by a slightly modified Oldendorf's method. 125I-Polyglobin was used as the control compound. The half-life of blood clearance of 125I-C-425 radioactivity after the 2nd day was about 8 days in rats and about 5 days in rabbits, respectively. Within a relatively short period of 1 hour, a rapid decrease of blood concentration of radioactivity was recorded. This was due to the transfer of labelled compound from blood into the extravascular compartment. A high concentration of radioactivity in the blood was maintained for one week after intravenous injection of the labelled compound in both rats and in rabbits. Most of the radioactive compound or metabolites were excreted in the urine and relatively little in feces. Excretion in urine of rats reached about 22% for 125I-C-425 and about 17% for 125I-Polyglobin. The excretion rate slowed down over 24 hours. The total recovery rates in rats were about 40% for 125I-C-425 and 125I-Polyglobin and about 55% in rabbits for both drugs. The permeability through the blood-brain barrier was found to be negligible for both drugs.

Animals↗
PubMed1994

Effect of various antidotes on the biliary and intestinal excretion of arsenic in situ and into the feces in vivo in guinea-pigs after injection of As2O3.

The effect of various antidotes on the excretion of arsenic into the feces in vivo and on the biliary and enteric excretion in situ was investigated on segments of jejunum and colon in anesthetized guinea-pigs using the pendular perfusion technique, according to Henning and Forth (1982). In the in situ experiments guinea-pigs received As2O3 (0.02 mmol As(III)/kg) and 30 min later, British-Anti-Lewisite (BAL), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA) or 2,3-bis-(acetylthio)propanesulfonamide (BAPSA) (0.1 or 0.7 mmol/kg each) into the jugular vein. In the in vivo experiments guinea-pigs received As2O3 s.c. (same dose as above) and 30 min later the same antidotes (0.1 mmol/kg i.p.). The feces were collected for 24 h and the arsenic content measured. During the 60-min perfusion period the amount of arsenic excreted into the jejunum or colon was only 3% or 0.4% of the dose administered, respectively. Of the arsenic dose, 8% was found in the bile. None of the antidotes had an effect on the arsenic excretion into the jejunum or colon. No change in biliary excretion was found in animals treated with BAL, 0.1 or 0.7 mmol/kg, respectively. DMSA, BAPSA or DMPS, 0.1 mmol/kg, increased the biliary excretion of arsenic to 14, 33, or 43% of the dose administered and after 0.7 mmol/kg to 29, 37, or 42%, respectively. Furthermore, a significant increase (P > 0.05) was found for the bile/blood concentration ratio in the following order: control < BAL < DMSA < BAPSA approximately DMPS.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

Recycling of nitrogen in herbivore feces: plant recovery, herbivore assimilation, soil retention, and leaching losses.

Herbivores directly and indirectly affect ecosystem functioning in forests. Feces deposition is a direct effect that supplies ephemeral N pulses to soils. Herbivore-mediated changes in plant N allocation and uptake are indirect effects that can also influence soil N availability. These effects may interact if defoliation influences the ability of plants to recover fecal N, and this may affect subsequent generations of herbivores. We added (15)N-enriched insect feces (frass) to a series of replicated red oak, Quercus rubra, mesocosms that had been damaged experimentally and then followed the frass N over the course of 2 years. In the first season, some frass N was mineralized in the soil and leached in organic form from the mesocosms within 1 week of deposition. Within 1 month, frass N had been acquired by the oaks and enriched the foliage; late-season herbivores assimilated the frass N within the same growing season. In the second season, herbivore damage from the previous year lowered total leaf N contents and (15)N recovered in the foliage. A subsequent cohort of early-season herbivores fed on this foliage consequently derived less of their N from the previous year's frass, and feral leaf rollers colonized fewer of these saplings. The 0- to 5-cm soil fraction was the largest N sink measured, and 42% of the frass N was recovered in the soil. The results demonstrate that: (1) some frass N can be recycled rapidly into foliage and assimilated by successive cohorts of herbivore within the same season; (2) damage can affect N allocation in the following year's foliage, influencing N availability to and host selection by herbivores; and (3) leaching losses occur soon after deposition but are buffered by soil pools, which are the largest sinks for frass N.

Analysis of Variance↗
PubMed2006

Identification of dominant bacteria in feces and colonic mucosa from healthy Spanish adults by culturing and by 16S rDNA sequence analysis.

The aim of this work was to examine by culturing the changes in the total and indicator populations of the feces of two individuals over 1 year and to identify the dominant microbial components of a single sample of feces from each donor. Populations and dominant bacteria from a sample of colonic mucosa from a further individual were also assessed. The culture results were then compared to those obtained with the same samples by 16S rDNA cloning and sequencing. High interindividual variation in representative microbial populations of the gastrointestinal tract (GIT) was revealed by both the culture and the culture-independent techniques. Species belonging to Clostridium clusters (XIVa, IV, and XVIII) predominated in both the fecal and the mucosal samples (except in the mucose cultured isolates), members of Clostridium coccoides cluster XIVa being the most numerous microorganisms. Species of gamma-proteobacteria (Escherichia coli and Shigella spp.), bifidobacteria, and actinobacteria appeared in lower numbers than those of clostridia. From the mucosal cultured sample, only facultative anaerobes and bifidobacteria were recovered, suggesting destruction of the anaerobe population during processing. In accordance with this, the microbial diversity revealed by 16S rDNA sequence analysis was greater than that revealed by culturing. Despite large interindividual differences, distinct human communities may have group-associated GIT microbiota characteristics, such as the low number of Bacteroides seen in the subjects in this study.

Adult↗
PubMed2006

Bifidobacterial diversity determined by culturing and by 16S rDNA sequence analysis in feces and mucosa from ten healthy Spanish adults.

This study aimed to identify the numerically predominant bifidobacterial species in feces and mucosa of healthy Spanish people and to determine their phenotypic and genetic diversity. To this end, both traditional culturing and molecular methods were used. A set of 196 bifidobacterial colonies was identified from the counting plates by sequencing of a stretch of their 16S rRNA gene. Representative isolates were phenotypically characterized by their carbohydrate fermentation profile and genotypically typed by RAPD-PCR. Four 16S rDNA libraries composed of 113 clones from two fecal and two mucosal samples were independently analyzed. Seven bifidobacterial species were identified by culturing, and six by 16S rDNA analysis. Both methodologies showed Bifidobacterium longum and B. pseudocatenulatum to predominate in feces and mucosa, although high interindividual variability was noted. High phenotypic variation was observed in the fermentation profile of different isolates from the same species. RAPD analysis showed that two to five strains made up the subjects' personal bifidobacterial communities. The identification of the dominant bifidobacterial species could be useful for the rational design, use, and evaluation of probiotics in our community.

Adult↗
PubMed2006

Foraging behavior of Anastrepha Ludens, A. obliqua, and A. serpentina in response to feces extracts containing host marking pheromone.

Following oviposition, females of many Tephritid flies deposit host marking pheromones (HMPs) to indicate that the host fruit has been occupied. We describe the foraging behavior of these three economically important species (Anastrepha ludens and A. obliqua from the fraterculus species group and A. serpentina from the serpentina species group) when they encounter an artificial fruit (green agar spheres wrapped in Parafilm) marked with intra- and interspecific feces extracts that contain, among other substances, host marking pheromone. When flies encountered fruit treated with either 1 or 100 mg/ml feces extract, there were drastic and statistically significant reductions in tree residence time, mean time spent on fruit, and in the number of oviposition attempts or actual ovipositions when compared to the control treatment (clean fruit). These responses were almost identical irrespective of extract origin (i.e., fly species), indicating complete interspecific HMP cross-recognition by all three Anastrepha species tested. We discuss the ecological and practical implications of our findings.

Animals↗
PubMed1975

A new convenient, rapid and accurate method for determination of total lipids in feces.

1. A new, simple and rapid colorimetric micromethod for determination of total lipids in the petroleum ether extract of feces is proposed. The method, based on the sulfophosphovanillin reaction, was found to be of satisfactory precision and accuracy. 2. The concentration of total lipids determined in meconium of 40 healthy newborns in the first five days of life, was 1 to 15 g/100 ml of feces.

Benzaldehydes↗
PubMed1980

Intestinal alkaline phosphatase: an immunoprecipitation method for the determination in feces.

An immunoprecipitation method for the determination of human intestinal alkaline phosphatase (I-AP) in feces is described using anti-human I-AP IgG. Antibodies to human I-AP are monospecific after absorption with human placental alkaline phosphatase and show no cross-reactions with human liver alkaline phosphatase or alkaline phosphatase from E. coli. Normal controls demonstrate a logarithmic normal distribution of I-AP activity in feces. The standard deviation of the intra- as well as of the inter-assay variation is 11.7%. This method represents a simple quantitative non-invasive in vivo assay for brush border damage under experimental and/or clinical conditions.

Alkaline Phosphatase↗