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PubMed2005

A new method for isolating colonocytes from naturally evacuated feces and its clinical application to colorectal cancer diagnosis.

BACKGROUND & AIMS: The early detection of colorectal cancer is desired because this cancer can be cured surgically if diagnosed early. The purpose of the present study was to determine the feasibility of a new methodology for isolating colonocytes from naturally evacuated feces, followed by cytology or molecular biology of the colonocytes to detect colorectal cancer originating from any part of the colorectum. METHODS: Several simulation studies were conducted to establish the optimal methods for retrieving colonocytes from any portion of feces. Colonocytes exfoliated into feces, which had been retrieved from 116 patients with colorectal cancer and 83 healthy volunteers, were analyzed. Part of the exfoliated colonocytes was examined cytologically, whereas the remainder was subjected to DNA analysis. The extracted DNA was examined for mutations of the APC, K-ras, and p53 genes using direct sequence analysis and was also subjected to microsatellite instability (MSI) analysis. RESULTS: In the DNA analysis, the overall sensitivity and specificity were 71% (82 of 116) of patients with colorectal cancer and 88% (73 of 83) of healthy volunteers. The sensitivity for Dukes A and B was 72% (44 of 61). Furthermore, the sensitivity for cancers on the right side of the colon was 57% (20 of 35). The detection rate for genetic alterations using our methodology was 86% (80 of 93) when the analysis was limited to cases in which genetic alterations were present in the cancer tissue. CONCLUSIONS: We have developed a new methodology for isolating colonocytes from feces. The present study describes a promising procedure for future clinical evaluations and the early detection of colorectal cancers, including right-side colon cancer.

Adult↗
PubMed1979

[Utilization of 15N-labelled urea by laying hens. 1. Survey of the literature, 15N-excretion in feces and urine].

Three colostomated leghorn hybrids with an average laying performance of 75% received a ration with 17.7% crude protein and an energy content of 519 energetic feed units for hens per kg mixed feed over a period of 8 days. In the first six days of the experiment the 1%-supplement of urea to the ration was labelled. Its atom-% 15N excess (15N') amounted to 96.06%. During the last two days the urea supplement was not labelled. The total N, trichloracetic acid (TCA)-soluble N and the ammonia N were determined in the feces samples collected daily. In the urine samples collected daily the total N, urea N and ammonia N per hen were determined as well. In all samples the atom-% 15N excess (15N') was measured. The percentage of 14N in feces of the 14N dose was, on an average of the three hens, 21.3% and the analogous quota of 15N' 4.6%. The quota of ammonia 14N of the total 14N in feces had an average of 2.5%, the corresponding 15N' quota was 10.1%. The atom-% 15N' of the urea N in urine was considerably above that of the total urine N and had a maximum of more than 50%. The quota of urine 14N of the 14N taken in had an average of 44.4%, and the corresponding 15N' quota was 56.9%. On an average of the three hens, 61.6% of the 15N' were excreted in feces and urine during the 8-day test period.

Ammonia↗
PubMed2003

Non-invasive screening for treatment of heifers with the anabolic steroid melengestrol acetate (MGA) by feces analysis.

For eight weeks, two heifers each had been orally administered daily doses of 0, 1.5, or 5 mg melengestrol acetate (MGA) in a feed premix. Four heifers received the labeled dose of 0.5 mg/day. Regular feces samples were taken throughout the experiment. A rapid screening method for the determination of MGA in feces was developed, consisting of liquid extraction, clean-up on solid-phase extraction cartridges and quantification by enzyme immunoassay (ELISA). Residues in feces were dose-dependent with mean values of < 0.25, 2.0, 4.4, or 15.4 ng/g for 0, 0.5, 1.5, and 5 mg/day doses, respectively. In contrast to urine analysis, feces analysis appeared to be a suitable means of non-invasive screening before slaughter for surveillance of MGA treatment and verification of its compliance with labeled dosage.

Administration, Oral↗
PubMed1991

Identification and quantitative distribution of eight analogues of naturally occurring fecapentaenes in human feces by high-performance liquid chromatography.

Fecapentaenes, highly potent fecal mutagens originating from intestinal bacterial production, have been suggested to play an essential role in the initiation of colorectal cancer. Reviewing the data on fecapentaene occurrence in man, the applied methodologies for fecapentaene extraction and analysis appear to be very inconsistent. Therefore, we compared several methods and developed an optimal extraction and purification procedure for fecapentaene quantification in human feces. This method is based upon a dichloromethane extraction of freeze-dried material with application of a Potter homogenization instrument and subsequent HPLC analysis in combination with photodiode array detection. This system enables us to detect and quantify at least eight forms of fecapentaene-like substances generally occurring in human stool. We suggest that these peaks represent fecapentaene-12 (FP-12) and fecapentaene-14, both with a geometric isomer, as well as fecapentaene analogues that have never been reported before. Applying this methodology on feces of a group of young healthy persons, we were able to detect fecapentaene levels ranging from less than 5 micrograms to 6 mg/kg feces, and in 40% of the samples greater than 1.0 mg/kg feces. The newly identified fecapentaenes represent 21.7% of total fecapentaene concentration. It appears that some fecapentaenes are excreted in higher amounts by females as compared to males. Furthermore, we found that fecal mutagenicity to Salmonella tester strain TA100 appeared lower than hypothesized on the basis of overall fecapentaene contents, and that fecal extracts diminish the mutagenic effect of synthetic FP-12 dramatically. Apparently, optimal conditions for fecapentaene extraction result also in an increased level of co-extracted anti-mutagenic substances. Determination of fecal mutagenicity as an index for fecapentaene excretion or colorectal cancer risk is therefore not suitable. In order to assess the relevance of fecapentaenes in the etiology of colorectal cancer, we suggest that a distinction should be made between relative occurrence and degree of genotoxic effect in situ of the various fecapentaene analogues.

Adult↗
PubMed2003

N-nitroso compounds in the gastrointestinal tract of rats and in the feces of mice with induced colitis or fed hot dogs or beef.

Because colonic N-nitroso compounds (NOC) may be a cause of colon cancer, we determined total NOC levels by Walters' method in the gastrointestinal tract and feces of rodents: (i) feces of C57BL mice fed chow and semi-purified diets contained 3.2 +/- 0.4 and 0.46 +/- 0.06 NOC/g, respectively (P < 0.01, mean +/- SD). (ii) NOC levels for gastrointestinal contents of three groups of Sprague-Dawley rats fed chow diet were 0.9 +/- 0.05 (diet), 0.2 +/- 0 (stomach), 0.3-0.4 (small intestine), 0.7-1.6 (cecum and colon) and 2.6 +/- 0.6 (feces) nmol/g. NOC precursor (NOCP) levels (measured as NOC after mild nitrosation) for two rat groups fed chow diet showed a 16-fold increase from stomach to proximal small intestine (mean, 6.2 micromol/g), and a 1.7-fold increase from distal colon to feces (mean, 11.6 micromol/g). (iii) Eight Min and five C57BL/6J mice received 4% dextran sulfate sodium in drinking water on days 1-4 to induce acute colitis. This increased fecal NOC levels 1.9-fold on day 5 in both strains (P < or = 0.04), probably due to NO synthase-derived nitrosating agents in the colon. (iv) Following studies on humans fed beef [Hughes et al. (2001) Carcinogenesis, 22, 199], Swiss mice received semi-purified diets mixed with 18% of beef plus pork hot dogs or sautéed beef for 7 days. On day 7, individual 24-h fecal NOC outputs were determined. In three hot dog and two beef groups with 5 mice/group, mean fecal NOC output/day was 3.7-5.0 (hot dog) and 2.0-2.9 (beef) times that for control groups fed semi-purified diet alone (P < 0.002 for each of combined groups). These groups showed little change in fecal NOCP output. (v) Initial purification of rat fecal NOCP by adsorption-desorption and HPLC is described. Results should help evaluate the view that colonic NOC causes colon cancer associated with colitis and ingestion of red and nitrite-preserved meat.

Animals↗
PubMed1979

Failure to detect hepatitis B surface antigen (HBsAg) in feces of HBsAg-positive persons.

Since previous studies did not provide conclusive data regarding the presence of hepatitis B surface antigen (HBsAg) in the feces of HBsAg carriers, the feces of 20 HBsAg carriers and six patients with HBsAg-positive active liver disease were examined with use of a reproducible method for concentrating and detecting HBsAg in feces in which bovine serum, which has been shown to protect HBsAg destruction by fecal components, was added to the fecal specimens. HBsAg was not detected in 66 fecal specimens from the 26 HBs-Ag-positive persons. Furthermore, HBsAg could not be detected in the feces of a carrier who had ingested 4 ml of his own serum. This finding suggests that there are factors in the gastrointestinal tract that interfere with the immunoreactivity of the HBsAg and possibly destroy the hepatitis B virus. These findings explain why oral/fecal spread does not play a major role in the transmission of hepatitis B virus.

Carrier State↗
PubMed1982

Diagnosis of giardiasis by counterimmunoelectrophoresis of feces.

The diagnosis of giardiasis is frequently difficult. In the present study, the techniques of microscopic examination of feces, duodenal fluid, and duodenal biopsy specimens were compared with counterimmunoelectrophoresis (CIE) of feces for Giardia lamblia antigen. New Zealand white rabbits were immunized with purified G. lamblia cysts and axial trophozoites, and the resulting antiserum was used for the detection of antigen. Of 276 patients with acute and chronic diarrhea studied using standard diagnostic methods, 66 patients had giardiasis: 62 by examination of feces; three, of duodenal fluid, and one, of duodenal biopsy specimens. CIE tests for G. lamblia fecal antigen had positive results for 65 of the 66 patients. The CIE test for Giardia fecal antigen appears to be as sensitive and reliable as the combined examination of feces and duodenal fluid.

Antigens↗
PubMed1998

Experimental infection of domestic cats with Bartonella henselae by inoculation of Ctenocephalides felis (Siphonaptera: Pulicidae) feces.

Caged cat fleas, Ctenocephalides felis (Bouché), were fed on 6 cats; 3 cats were injected with 5 x 10(7) colony forming units of Bartonella henselae intradermally and 3 cats were injected with an equal volume of saline. After the fleas fed for 4 d, 5 groups of 50 B. henselae-exposed fleas were caged and allowed to feed on 5 cats for 6 d. Five cats each were injected intradermally with 1 ml of saline containing 45 mg of feces from B. henselae-exposed fleas. Five cats were fed 50 B. henselae-exposed fleas and 45 mg of fresh feces from B. henselae-exposed fleas. Five cats received all 3 treatments by using fleas and feces collected from cats inoculated with saline (controls). Cats were bled weekly and tested by culture and serology. The cats that were injected with feces from infected fleas were positive by culture for B. henselae at 1 or 2 wk after exposure and were the only cats to become bacteremic or seropositive by week 20.

Animals↗
PubMed1976

Enzymic activities in the pancreas, digestive tract and feces of rats fed raw or heated soy flour.

Trypsin, chymotrypsin and amylase levels were determined in the pancreas, all along the intestinal tract, and in the feces of rats fed raw or heated soy flour diets. The levels of all enzymes measured in the pancreas in the non-fasted state were lower in the raw than in the heated soy flour-fed rats. Fasting equalized these levels. Trypsin and amylase tended to be lower, and chymotrypsin was significantly higher in the intestinal tracts of raw soy flour-fed rats than in the group fed heated soy flour; the greatest differences were found in the ileum. Trypsin and chymotrypsin levels in the feces were higher in the group fed raw soy flour than in the group fed heated soy flour. Amylase in the feces of the raw soy flour-fed rats was higher at the beginning of the experiment and dropped sharply to be even lower than in the heated soy flour-fed rats at days 13 to 14 of the experiment. It was concluded that measuring the enzymic levels in the feces is a very sensitive method for determining whether a test diet induces hypersecretion of digestive enzymes in rats. This method can be used from the start of feeding the experimental diet. As the animal need not be killed, the effect of the test diet upon enzymic secretion can be studied as a function of time, and it might be suitable to studies with large animals.

Amylases↗
PubMed1997

Consumption of retrograded (RS3) but not uncooked (RS2) resistant starch shifts nitrogen excretion from urine to feces in cannulated piglets.

To study the effect of resistant starch (RS) on the route of nitrogen excretion, we fed three groups of six cannulated piglets each a diet containing either uncooked resistant starch (RS2 ), retrograded resistant starch (RS3 ) or glucose. The use of piglets with a cannula at the end of the ileum allowed measurement of the amount of nitrogen that entered the colon. Ileal digesta, urine and feces were collected quantitatively and weighed, and dry matter, starch and nitrogen content were determined. We hypothesized that RS2 would lower colonic absorption of nitrogen when compared with RS3 , because RS2 may be more fermentable than RS3 , thus trapping more nitrogen in bacteria. The piglets fed RS3 had a significantly higher production of ileal digesta and feces than the piglets fed glucose or RS2 . In the piglets fed RS2 , 44% of the amount of RS fed was recovered in the ileal digesta; in the piglets fed RS3 , 71% was recovered. Thus, more fermentable material entered the colon in the RS3 -fed piglets than in the RS2 -fed piglets. Virtually no starch was recovered in the feces of any dietary group. Replacement of glucose by either RS2 or RS3 did not affect nitrogen retention but increased fecal nitrogen excretion. Compared with glucose, RS3 but not RS2 reduced urinary nitrogen excretion, mainly in the form of urea, and reduced the amount of nitrogen absorbed by the colon when expressed as a percentage of the amount of nitrogen entering the colon. This study provides evidence that RS3 , but not RS2 , shifts nitrogen excretion from urine to feces in cannulated piglets.

Animals↗
PubMed2004

Recovery of Campylobacter from broiler feces during extended storage of transport cages.

Feces deposited in transport cages by a Campylobacter-positive flock can cause the spread of Campylobacter to subsequent flocks placed in the same cages. This experiment was designed to determine the effect of extended cage storage on the viability of Campylobacter in feces deposited on the cage floor during commercial transport and holding. After 4 h of feed (but not water) withdrawal, Campylobacter-positive broilers were caught by commercial catching crews, placed into 3 new commercial cages and transported with the rest of the flock to the holding area at a commercial processing facility. Broilers were allowed to remain in the cages for 8 h before being unloaded by facility personnel. After removal of the broilers, empty cages were held under a shed and sampled at 7 intervals for the presence of viable Campylobacter. Cages were sampled by removing all the feces out of a different randomly assigned compartment in each cage at 0.5, 2, 4, 6, 8, 24, and 48 h after unloading. No decrease in Campylobacter numbers was noted through 8 h of storage. After 24 h in both replications, Campylobacter was detected in 2 of 3 compartments by direct plating and detected in the third by enrichment only. After 48 h, Campylobacter was detected in one replication by enrichment only, and was not detected in the second replication at all. Storing soiled transport cages for 48 h between uses results in lower numbers of Campylobacter in feces, but may not eliminate Campylobacter entirely. Due to cage cost and space requirements, routine cage storage between uses would not be practical.

Animals↗
PubMed2006

Quantification of [3H]docetaxel in feces and urine: development and validation of a combustion method.

Most radiolabeled biological samples require extensive sample preparation to reduce quenching interference before quantification of radioactivity is possible. Clearly, a more rapid and simple method ensuring a constant count rate and optimal counting efficiency has important advantages. We report on the development and analytical method validation of a rapid and simple combustion method to quantify [3H]docetaxel excreted in human feces and urine. A 3-day validation procedure was performed; quality control (QC) samples, prepared in blank feces and urine, were combusted 5 times and aliquots of the produced tritiated combustion water were counted in a liquid scintillation counter. The validation runs demonstrated adequate precision (below 7.6%) across all QC levels. Sensitivity at the lowest QC level was excellent and recovery of radioactivity constant (ranging from 85 to 91.8%). Clinical applicability of the method was tested in a cancer patient receiving docetaxel and a tracer amount of [3H]docetaxel; during the first 72 h after [3H]docetaxel infusion, 60% of total radioactivity was excreted in the collected feces and urine, which is within the expected range. Combustion of tritiated feces and urine samples is a simple, rapid, sensitive, precise and reproducible method with high recovery. It can be applied to quantify [3H]docetaxel excretion after i.v. administration.

Antineoplastic Agents, Phytogenic↗
PubMed2004

Detection of Helicobacter pylori DNA in feces and saliva by polymerase chain reaction: a review.

The polymerase chain reaction (PCR), known for its high sensitivity and specificity, has been used for the detection of Helicobacter pylori DNA in bodily materials such as feces and saliva. Since fecal specimens contain PCR inhibitors, DNA before PCR amplification has been purified using various biochemical, immunological and physical pre-PCR steps. Several PCR protocols, differing from each other in the selection of genomic targets and primers, have produced varying degrees of specificity and sensitivity in detecting H. pylori DNA. PCR identified antimicrobial resistance of H. pylori in feces. It also detected virulence factor genes such as the cytotoxin-associated gene (cagA) and vacuolating cytotoxin gene (vacA) in feces and saliva. While the cagA gene was detected in 50-60% of fecal specimens, it was found in 25% of salivary specimens from patients. There was considerable variation in the detection rate of H. pylori DNA in salivary samples. The detection rate in saliva with the most effective primer pair was lower than that observed in feces, making saliva a less suitable specimen for the diagnosis of H. pylori infection. There is controversy regarding the permanent presence of H. pylori in saliva. Whether the salivary and gastric specimens of an individual harbor identical or different strains has not been resolved. PCR cannot distinguish between living and dead organisms. However, it can offer quick results on fecal and salivary specimens, which may contain fastidious and slow-growing H. pylori in low numbers.

Antigens, Bacterial↗
PubMed2005

Peroral infectivity of Toxoplasma gondii in bile and feces of interferon-gamma knockout mice.

Toxoplasama gondii appeared in the bile and feces of interferon-gamma knockout (GKO) but not wild type mice on days 7-8 after peroral infection with T. gondii cysts of Fukaya strain. Both tachyzoite-specific SAG1 and bradyzoite-specific T.g. HSP30 mRNAs were detected in the bile and feces of GKO mice. Tachyzoites converted to bradyzoites by culturing in the bile. By feeding uninfected mice with the bile and washed feces of T. gondii-infected GKO mice, T. gondii-specific antibody formation in the serum and cyst formation in the brain were observed. The novel migration route of T. gondii from liver to bile and feces in GKO mice was confirmed.

Animals↗
PubMed1996

In vitro degradation of oxalic acid by human feces.

BACKGROUND: Oxalic acid-degrading bacteria have been isolated from human faces. Therefore, the possibility exists that oxalic acid in food is degraded in human intestine by such bacteria, and absorption and excretion of oxalic acid is reduced. It may be possible that patients who form idiopathic stones have fewer oxalic acid-degrading bacteria than healthy controls. The purpose of this study is to examine the possibility that oxalic acid in food is degraded in the human intestine. METHODS: Nineteen patients with calcium stones and 13 healthy subjects were included in the study. Samples of feces were diluted with Barber medium containing 1 g/L of oxalic acid dihydrate, and 1%, 0.1% and 0.01% suspensions were prepared. These solutions were incubated under anaerobic conditions at 37 degrees C for seven days. The degradation of oxalic acid was estimated by the decrease of oxalic acid in the medium. RESULTS: The feces of almost all persons with or without calculi degraded oxalic acid to some degree. Incomplete or no oxalic acid degradation was found in 15 of the 19 stone-forming patients and in five of the 13 stone-free controls. CONCLUSION: Large numbers of oxalic acid-degrading bacteria were observed less often in the feces of stone-formers than in the feces of stone-free controls.

Adult↗
PubMed2006

Helicobacter anseris sp. nov. and Helicobacter brantae sp. nov., isolated from feces of resident Canada geese in the greater Boston area.

Numbers of nonmigratory Canada geese have increased substantially in the past decade, and they have become a nuisance in some urban areas. Because of their close contact with humans in parks and areas adjacent to surface waterways, contact with their feces poses a zoonotic risk. A total of 97 geese from 10 separate geographic locales in the greater Boston area had their feces sampled for detection of Helicobacter spp. Identification of Helicobacter spp. based on 16S rRNA genus-specific helicobacter primers was noted in 39 of 97 (40.2%) DNA fecal extracts. Twenty-seven (27.8%) of these geese had helicobacters isolated from their feces. A urease-positive novel species, Helicobacter anseris, based on phenotypic, biochemical, and 16S rRNA analyses, was isolated from 20 geese from seven different flocks. A second, novel, urease-negative Helicobacter sp., H. brantae, was identified in seven geese. Four geese had both novel Helicobacter spp. cultured from their feces. Whether these two novel helicobacters pose a zoonotic risk, similar to other enteric helicobacters (e.g., H. canadensis, previously isolated from diarrheic and bacteremic humans and from geese in Europe), will require further studies.

Animals↗
PubMed1986

Enumeration of polysaccharide-degrading Bacteroides species in human feces by using species-specific DNA probes.

DNA probes that are specific for each of five predominant species of human colonic Bacteroides (B. thetaiotaomicron, B. uniformis, B. distasonis, "Bacteroides group 3452-A", and B. ovatus) were used to detect and enumerate these species in fecal samples from two adult volunteers. These five species are capable of fermenting many of the complex polysaccharides that are thought to be sources of carbon and energy for bacteria in the colon. Estimates for the concentrations of some of these species in feces have not been previously available because of the difficulties in differentiating colonic Bacteroides spp. by conventional biochemical tests. Our results indicate that all the species except B. ovatus were present in high numbers (greater than 10(9)/g [dry weight]) in the feces of both volunteers. However, the concentrations of the more versatile polysaccharide-degrading species within this group of organisms (7.6 X 10(9) to 12.0 X 10(9)/g [dry weight] for B. thetaiotaomicron; 2.9 X 10(9) to 6.3 X 10(9)/g [dry weight] for "Bacteroides group 3452-A") did not differ significantly from the concentrations of less versatile polysaccharide-degrading species (1.2 X 10(10) to 2.0 X 10(10)/g [dry weight] for B. uniformis; 5.8 X 10(9) to 8.4 X 10(9)/g [dry weight] for B. distasonis). B. ovatus was not detectable by our method. Since our lower limit of detection is approximately 1 X 10(9) to 2 X 10(9)/g (dry weight) of feces, this is consistent with earlier estimates that indicated that the concentration of B. ovatus in feces is near or below this value.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteroides↗
PubMed1990

Neutral sugar composition and gravimetric yield of plant and bacterial fractions of feces.

Separating dietary fiber from other polysaccharides in digesta and feces is necessary to understand its mechanisms of action. A gravimetric method that separates fecal plant and bacterial matter based on size and density was evaluated and modified to determine the plant and bacterial mass of lyophilized whole and blended rat and human feces. Three screen mech combinations (150 and 75 microns, 150 and 35 microns, 35 microns) were used with rat feces. Filtration of a homogenized rat fecal slurry sequentially through 150- and 35-microns-mesh screens versus 150- and 75-microns-mesh screens decreased the gravimetric recovery of bacteria from congruent to 35 to congruent to 25% of fecal dry weight and increased the plant fraction weight. Neutral sugar composition, determined by gas chromatography of alditol acetates, and bacterial counts of the fractions suggested that the decreased yield of bacterial fraction represented removal of plant material and not a loss of bacteria. Rat excreta contained 29.5% (dry weight) total neutral sugar, 88% of which was recovered in the plant material. Human feces containing wheat bran, fractionated with the 150- and 35-microns-mesh screens, was 21% neutral sugar, congruent to 65% of which was in the plant fraction. The plant fractions had more xylose and arabinose and less glucose than the bacterial fractions. Processing samples in a Waring blender had no adverse effect on the rat or human fecal bacterial counts. The use of this gravimetric method in combination with the sugar analysis of the fractions provided a better measure of plant and bacteria than only gravimetric yield.

Animals↗