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PubMed1993

Role of gastric pH in isolation of Helicobacter mustelae from the feces of ferrets.

BACKGROUND: Helicobacter mustelae colonizes the gastric mucosa of ferrets and causes persistent chronic gastritis. METHODS: Hypochlorhydria, as measured by gastric pH probe, was induced by administering oral omeprazole, a proton pump inhibitor of the parietal cell, to adult ferrets in two separate experiments. Feces of ferrets were cultured for H. mustelae before, during, and after omeprazole therapy. RESULTS: H. mustelae was isolated in 23 of 55 (41.8%) sequential fecal samples collected during omeprazole therapy. The same ferrets with acidic gastric pH had H. mustelae isolated in 6 of 62 (9.7%) of the fecal cultures (P < 0.01). A DNA species-specific H. mustelae probe confirmed the presence of the organism in both the stomach and feces of all 5 ferrets. In 4 of 5 ferrets restriction enzyme patterns of the gastric H. mustelae were identical to those of the fecal H. mustelae strains. CONCLUSIONS: Hypochlorhydria promotes fecal transmission of a gastric Helicobacter organism. The H. mustelae-colonized ferret provides an ideal model to study the epidemiology and pathogenesis of Helicobacter pylori-induced gastritis.

Animals↗
PubMed1981

Inhibitory effect of the ether extract of human feces on activities of mutagens: inhibition by oleic and linoleic acids.

An ether extract of normal human feces showed inhibitory effects on the activities of several mutagens in the Ames tests. By addition of the ether extract at an amount equivalent to 0.5 g of a sample of feces, the mutagenicity of 1.5 nmole of 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) on Salmonella typhimurium TA98 was completely inhibited. No killing of the bacteria was detected during this treatment. Other mutagens also subject to the inhibition were 2-amino-6-methyl-dipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1), 2-amino-9H-pyrido[2,3-b]indole (Glob-P-2), 2-amino-3-methylimidazo[4,5-d]quinoline (IQ), benzo[a]pyrene and aflatoxin B1. Apart from these mutagens, which require S9 for their activation, the direct mutagen prepared from Trp-P-1 by treatment with S9 was also inhibited by the fecal extract. The inhibitory principles in the fecal extract were fractionated by thin-layer chromatography on silica gel and were identified as oleic and linoleic acids. Whereas these unsaturated fatty acids showed strong inhibitory activities, saturated fatty acids, i.e, stearic and palmitic acids, did not exhibit any inhibition. Although the physiological significance of these effects of oleate and linoleate is yet to be elucidated, this finding has indicated that care must be taken in screening mutagens by the Ames tests to avoid false negatives resulting from the presence of unsaturated fatty acids in the system.

Feces↗
PubMed1988

Food intake suppression in rats by a substance isolated from human feces.

A substance which suppresses food intake was isolated from human feces. This substance was extracted and partially purified using Sephacryl S-200 column chromatography and a DEAE-Sephadex A-25 column chromatography. To assess the biological activity, rats were subjected to various applications. The intraperitoneal administration and the intracerebroventricular microinfusion of this substance suppresses short- and long-term food intake. The central infusion suppresses feeding with a potency over 1000 times that the peripheral administration. The central infusion of the heat-treated substance was without effect. These results suggest that a substance present in human feces (probably of a proteic nature) suppresses food intake in rats, at the level of the central nervous system.

Animals↗
PubMed1983

Vitamin K content of liver and feces from vitamin K-deficient and butylated hydroxytoluene (BHT)-treated male rats.

Vitamin K content of liver and feces from male rats fed diets containing butylated hydroxytoluene (BHT) was estimated by a chick bioassay method to investigate the mechanism of BHT-induced decreases in the activities of vitamin K-dependent clotting factors. The concentration of vitamin K in the liver of rats receiving BHT was reduced as compared to that of control rats. Conversely, the concentration of vitamin K in the feces from rats receiving BHT increased more than that from control rats. The vitamin K deficiency induced by BHT might be due to effects of BHT on absorption and excretion of vitamin K.

Animals↗
PubMed1983

Mutagenicity of different fractions of extracts of human feces.

An extraction-fractionation scheme for the isolation of non-volatile fecal mutagens is described. Extraction of feces was with acetone, and a 2-step fractionation scheme employing silica gel SepPak cartridges and normal-phase high-pressure liquid chromatography was used. Assay of mutagens was with the standard plate mutagenicity assay with Salmonella typhimurium tester strains TA98 and TA100, with and without the Aroclor induced S9 microsomal activation system. Single feces samples from 24 donors from a wide socioeconomic spectrum were tested. It was found that most fecal mutagenicity extracted by aqueous acetone could be attributed to a lipid soluble mutagen active on both TA98 and TA100 that has been previously reported.

Chromatography↗
PubMed1992

A rapid method for detection of Yersinia enterocolitica serotype O:3 in pig feces using monoclonal antibodies.

A simple colony immunoblotting method using monoclonal antibodies (MAbs) was developed to detect Y. enterocolitica serotype O:3 in pig feces. One of the MAbs studied was able to detect single colonies of the organism in the presence of calculated 3.1 x 10(8) heterologous organisms in pig feces. The MAb was found to be specific for the lipopolysaccharide (LPS) O-antigens of Y. enterocolitica serotype O:3. No significant cross-reactivity was found against a variety of closely related serotypes and Gram-negative organisms. The MAb could also be used in a slide agglutination test and an indirect fluorescence antibody assay for rapid identification of Y. enterocolitica serotype O:3.

Agglutination Tests↗
PubMed1985

63Ni electron-capture gas chromatographic assay for buprenorphine and metabolites in human urine and feces.

A 63Ni electron-capture gas chromatographic assay is described for buprenorphine, a potent narcotic agonist--antagonist. In addition, the assay is useful for the measurement of the metabolite norbuprenorphine and demethoxybuprenorphine, a rearrangement product resulting when buprenorphine is exposed to acid and heat. An extraction procedure was developed which optimized recovery of buprenorphine from biological samples and produced minimal background interferences and emulsion problems. Extract residues were derivatized with pentafluoropropionic anhydride and assayed by gas chromatography. Samples were analyzed with and without enzyme hydrolysis, thus providing a selective and sensitive assay for both free and conjugated buprenorphine, norbuprenorphine and demethoxybuprenorphine. The lower limits of detection following extraction of a 1-ml sample were ca. 10 ng/ml for buprenorphine and demethoxybuprenorphine and 5 ng/ml for norbuprenorphine. Application of the assay to human samples following a 40-mg oral dose of buprenorphine produced no evidence for the presence of demethoxybuprenorphine in urine or feces. Norbuprenorphine (free and conjugated) was present in urinary and fecal samples; buprenorphine (free and conjugated) was found in high amounts only in feces and in trace amounts in urine as conjugated buprenorphine. The urinary and fecal excretion pattern observed for a human subject following oral dosing of buprenorphine suggests enterohepatic circulation of buprenorphine.

Adult↗
PubMed1987

Comparison of electron microscopy with three commercial tests for the detection of rotavirus in animal feces.

Three commercial test kits were evaluated to detect the presence of rotavirus antigens in bovine, porcine, and turkey feces. Two of the assays, Rotalex (Medical Technology Corporation, Somerset, NJ) and Virogen-Rotatest (Wampole Labs, Cranbury, NJ) are latex agglutination tests (LA), while the third, Pathfinder (Kallestad, Austin, TX) is an enzyme immunoassay. The clinical usefulness of these assays was elevated by comparing their results with those of direct electron microscopy (EM). A total of 135, 92, and 211 samples of animal feces were tested by Rotalex, Virogen, and Pathfinder, respectively. All samples were examined by EM as a reference procedure. The overall agreement of the three commercial assays with EM was 53%, 66%, and 83% for Rotalex, Virogen, and Pathfinder, respectively. Based on these results, we consider Pathfinder as an attractive alternative to EM for the detection of rotavirus in animals. Of the two LA tests, Virogen was found to be a little more sensitive and specific.

Animals↗
PubMed2005

Enzyme immunoassay of progesterone in the feces from beef cattle to monitor the ovarian cycle.

The present study was undertaken to measure fecal progesterone concentration of beef cattle using antibody against authentic progesterone and to examine whether this method can monitor the ovarian cycle in beef cattle. Rectal fecal samples collected from 14 beef cattle were mixed with 6 ml of 100% methanol and shaken for 15 min. After centrifugation, supernatant was extracted with petroleum ether followed by an enzyme immunoassay (EIA) for progesterone. Specificity of the assay was examined by HPLC separation of fecal solution followed by the EIA in each fraction. The present assay identified only progesterone but not other metabolites in the feces sample that was extracted with petroleum ether. Sensitivity of the assay was estimated to be 0.0055 ng/ml (0.11 ng/g). Coefficient variations of intra- and inter-assay were 9.6-10.9% and 10.8-16.6%, respectively. Recovery rates ranged between 73 and 84%. Patterns in the fecal progesterone concentrations during the ovarian cycle were almost parallel to the plasma concentrations. A significant positive correlation was established between the fecal and plasma progesterone concentrations in individual animal (r=0.59-0.84, P<0.001, n=10) as well as pooled data (r=0.70, P<0.001, n=65). Fecal progesterone concentrations of day 0 (showing the nadir of concentration) of the ovarian cycle were less than 50 ng/g, which increased significantly toward day 9 (P<0.01). From days 14 to 18, there was significant reduction of fecal progesterone concentration (P<0.01). Ovarian cycles had at least 48 ng/g (mean=74 ng/g) of difference between minimum and maximum fecal progesterone concentrations. All cattle at days 9, 11 and 14 had higher fecal progesterone concentrations by more than 20 ng/g compared with day 0. These results suggest that the present EIA is suitable to measure the progesterone in cattle feces and can monitor ovarian cycle.

Animals↗
PubMed2004

Non-invasive diagnosis of Helicobacter pylori infection: simplified 13C-urea breath test, stool antigen testing, or DNA PCR in human feces in a clinical laboratory setting?

OBJECTIVES: (1) To compare two stool antigen EIAs (HpSA, FemtoLab) and PCR of ureaseA and cagA in feces, with (13)C-urea breath test (UBT). (2) To ascertain whether a simplified UBT (breath collection time = 10 min) is as reliable as the standard assay (30 min). DESIGN AND METHODS: Helicobacter pylori status was recorded in Group 1 (n = 187) by UBT, H. pylori stool antigen, ureA and cagA PCR in feces. UBT with 10, 20 and 30 min sampling was performed in Group 2 patients (n = 283). RESULTS: The sensitivity and specificity of HpSA, FemtoLab, and ureA were 67% and 99%, 90% and 96%, 35% and 98%, respectively. cagA results were positive in 16/48 H. pylori-positive, and in 5/100 H. pylori-negative patients. The results of UBT with a 10- and 30-min sampling strictly overlapped. CONCLUSION: UBT with 10 min breath collection and FemtoLab stool antigen assay are the most reliable non-invasive tests to diagnose H. pylori infection.

Adult↗
PubMed2004

High-performance liquid chromatographic assay of lactic, pyruvic and acetic acids and lactic acid stereoisomers in calf feces, rumen fluid and urine.

To facilitate clinical investigation of metabolic acidosis, a high-performance liquid chromatographic method was adapted and validated for the chiral separation of D-(-) and L-(+)-lactic acid in calf feces, rumen fluid and urine. A non-chiral method was also adapted and validated for the separation of pyruvic, acetic and DL-(+/-)-lactic acids in calf feces and DL-(+/-)-lactic and pyruvic acids in rumen fluid. Separation and quantification were achieved using a reversed phase sulphonated polystyrenedivinylbenzene analytical column for pyruvic, acetic and racemic lactic acids and by a 3 microm octadecylsilane (ODS) packed analytical column coated with N,N-dioctyl-L-alanine as the chiral selector for the separation of lactic acid enantiomers with Cu(II)-containing eluents by stereoselective ligand exchange chromatography. Endogenous analytes were present in validation samples over a range of concentrations (0.2-14.8 mmol/l). For the stereoselective assay, mean intra-day accuracy ranged from 90.6 to 108.4% and intra-day precision from 0.3 to 13.8%. For the non-stereoselective assay, mean intra-day accuracy ranged from 90.4 to 108.8% and intra-day precision from 1.5 to 11.1%. The limit of quantitation was 1.0 mmol/l for D- and L-lactic acid, 0.06125 mmol/l for pyruvic acid, 1.0 mmol/l for DL-lactic acid and 1 mmol/l for acetic acid. These assays can be used to study the role of the gastrointestinal tract and kidney in metabolic acidosis.

Acetic Acid↗
PubMed2004

Development of multi-color FISH method for analysis of seven Bifidobacterium species in human feces.

We have developed a multi-color fluorescence in situ hybridization (FISH) method which detects, by a single reaction, all seven species of Bifidobacterium (B. adolescentis, B. angulatum, B. bifidum, B. breve, B. catenulatum, B. dentium, and B. longum), the dominant bacteria in human feces. First, eight new types of oligonucleotide probe were designed, complementary with the 16S rRNA sequence specific to genus Bifidobacterium and each bifidobacterial species described above. Using whole cell hybridization, the fluorescent intensity was measured against the bacterial species targeted by each probe, to show that each probe is specific to the targeted bacteria and that the relative fluorescent intensity (RFI) as an indicator of probe accessibility is high at 61-117%. Then, bacterial species-specific probes were labeled with fluorochromes (FITC, TAMRA and Cy5) in seven different ways, singly or in combination. Using these probes, seven species of Bifidobacterium were differentially stained in mixed samples of cultured bacteria and feces from adult volunteers, proving the efficacy of this technique.

Bifidobacterium↗
PubMed2005

Quantification of bacteria in human feces using 16S rRNA-hybridization, DNA-staining and flow cytometry.

Hybridization of bacteria with fluorescent probes targeting 16S rRNA and inspection of hybridized bacteria with fluorescence microscopy (microscopy-FISH, i.e. fluorescence in situ hybridization) have constituted an accessible method for the analysis of mixed bacterial samples such as feces. However, microscopy-FISH is a slow method and prone to errors. Flow cytometry (FCM) enables analysis of bacteria more rapidly, accurately and reliably than microscopy. In this study, a FCM method for the analysis of 16S rRNA-hybridized and DNA-stained fecal bacteria was developed. The results of FCM-FISH were comparable to those of microscopy-FISH, and the coefficients of variation of the FCM analyses were extraordinarily low. In previous FCM-FISH studies, the Eub 338 probe, which is supposed to hybridize all bacteria, has been used to detect all bacteria present in the sample. We found that Eub 338 did not bind to all bacteria, which could be detected by DNA-staining; while SYTOX Orange DNA-stain detected all bacterial species tested and produced high fluorescence intensities enabling clear separation of bacteria from non-bacterial material. Thus, DNA-staining is a method of choice for the detection of all bacteria in FCM-FISH. We conclude that FCM of 16S rRNA-hybridized and DNA-stained bacteria is a rapid and reliable method for the analysis of mixed bacterial samples including feces.

Actinobacteria↗
PubMed2005

Isolation of tannin-degrading bacteria isolated from feces of the Japanese large wood mouse, Apodemus speciosus, feeding on tannin-rich acorns.

Bacteria with tannase activity were isolated from the feces of the Japanese large wood mouse, Apodemus speciosus. They were largely classified into two groups: Gram-positive cocci and Gram-positive bacilli. Genotypic analysis using a species-specific PCR assay as well as biochemical tests identified all cocci as Streptococcus gallolyticus. A PCR assay targeting a genus-specific sequence in the 16S/23S rDNA spacer region and additional 16S rDNA sequencing indicated that the bacilli belong to the genus Lactobacillus, with L. animalis and L. murinus being closely related taxa. Subsequent estimation of guanine-plus-cytosine content, amplified ribosomal DNA restriction analysis, and DNA/ DNA hybridization assay confirmed that the bacilli are homologous to each other but different from L. animalis or L. murinus. Consequently, a novel species of the genus Lactobacillus may be proposed. To date, this study is the first to report on the isolation of tannase-positive bacteria from the feces of a rodent species. These bacteria may play an essential role for the host organism in digesting tannin-rich acorns available in their natural habitats, thereby endowing them with a greater ecological advantage.

Animals↗
PubMed2004

Temperature effect on aerobic biodegradation of feces using sawdust as a matrix.

Temperature is one of the most important factors affecting microbial growth and biological reactions. In this study, the effect of temperature on aerobic biodegradation of feces is described through the comparison and analysis of experimental oxygen utilization rates (OUR) profiles obtained from batch tests conducted at several temperatures covering mainly mesophilic and thermophilic ranges. Additionally, the temperature effect was incorporated into the bio-kinetic model introduced by Lopez Zavala et al. (Water Res 38(5) (2004) 1327) and simulation of experimental OUR profiles was conducted. Results show that mesophilic and thermophilic microorganisms behaved differently to temperature; additionally, results suggest that the optimum temperature from the viewpoint of feces biodegradability is within the thermophilic range, nearly 60 degrees C. The enzymatic activity of microorganisms at 70 degrees C was remarkably diminished. For better predictions in the mesophilic range, two fractions of slowly biodegradable organic matter were identified, easily hydrolyzable organic matter (X(Se)) and slowly hydrolyzable organic matter (X(Ss)).

Bacteria, Aerobic↗
PubMed2004

Evaluation of adrenal function in serum and feces of Steller sea lions (Eumetopias jubatus): influences of molt, gender, sample storage, and age on glucocorticoid metabolism.

Fecal corticosterone concentrations, measured via radioimmunoassay (RIA), were validated as a method to monitor adrenal function in Steller sea lion physiology. Quantification of adrenal response to an acute stressor and relevance of data produced by developed methodologies was determined through physiological challenge with exogenous administration of adrenocorticotropic hormone (ACTH) to captive adult, reproductively intact, Steller sea lions of both sexes (n=3, 1 male, 2 female) during seasonal molt. Following ACTH administration, serial blood and fecal samples were collected and analyzed by RIA to determine adrenal response. Storage regimens and weather exposure were examined to establish external impact on fecal corticosterone concentrations. High-pressure liquid chromatography (HPLC) of both serum and feces of Steller sea lions was employed to explore potential gender-based differences extant in either sample media. ACTH challenges produced >3-fold increases in serum cortisol concentrations which were reflected in >18-fold increases in fecal corticosterone concentrations post-injection at 3.25 and 32 h, respectively, and fecal corticosterone concentrations returned to baseline 52 h post-injection. Neither outdoor exposure to weather nor variation in duration and temperature of freezer storage impacted fecal corticosterone concentrations. HPLC of individual fecal samples produced eluate immunoreactivity profiles that differed consistently with both sex and age class. Techniques developed herein effectively detected physiologically relevant corticosterone data in Steller sea lion feces, unaffected by conditions likely to be encountered with field collection samples. Additionally, results quantify an acute response to ACTH and provide methodology for examining chronically heightened adrenal activity in Steller sea lions.

Adrenal Glands↗
PubMed1990

Determination of [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin in human feces to ascertain its relative metabolism in man.

Human feces samples from a self-dosing experiment were analyzed by gas chromatography/mass spectrometry (GC/MS) for [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin (3H-2378-TCDD) to determine that 36-44% of the radioactivity was attributable to the parent compound. This method, using isotope dilution analysis, proved to be difficult due to the unexpectedly higher native 2378-TCDD background which created abnormally large precision ranges around the calculated feces concentrations of 0.1-0.2 pg/g. These results were supported by additional analyses involving the GC/MS chemical cleanup method combined with liquid scintillation counting which showed that at most, 50% of the radioactivity was due to 2378-TCDD metabolites resulting in a minimum metabolism of 50% for these samples.

Dioxins↗
PubMed2005

Effects of resistant potato starch on odor emission from feces in Swine production units.

Odor emission from swine facilities is determined by microbial breakdown of amino acids or carbohydrates in the pig colon. It was the aim to influence apoptosis and thus amino acid availability for odor formation by feeding resistant starch (300 g kg(-1) feed) over the whole fattening period to 40 pigs. Concentrations of 12 key components (indoles, volatile fatty acids, methanethiol) were measured in feces and headspace over the slurry duct and compared to 40 normally fed controls in a separate compartment. Concentrations of substances resulting from amino acids were reduced in feces by 70% (indoles) and 8% (branched chain fatty acids) and in the headspace by 72% and 20%. Resistant starch only led to minor increases of straight chain fatty acid concentration. Maximal reduction occurred for 3-methyl-1H-indole (skatole) which is the main determinant of malodor so that the results point to promising strategies for reducing pig odor emission.

Animals↗