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Fecal odorgrams. A method for partial reconstruction of ancient and modern diets.

Modern and ancient human feces (coprolites) and modern nonhuman animal fecal samples were subjected to headspace gas-chromatographic analysis for detection of odors related to dietary intake. The odors of grass, green leaves, licorice, meat, and corn were detected in coprolites dating to 6400 years BP (before present). Analysis of modern human samples revealed a large number of odors reflecting dietary constituents. The complexity and number of fecal dietary odors recovered was directly related to the number of food items ingested. Animal fecal odorgrams were characterized by a relatively large number of pleasant odors. Fecal odorgrams represent a new method by which modern and ancient diets may be partially reconstructed.

Animals↗

Passive immunization against somatostatin increases resistance to Eimeria vermiformis infection in susceptible mice.

The effect of in vivo immunoneutralization of somatostatin (SRIF) on Eimeria vermiformis intestinal infection was studied in resistant (BALB/c), and susceptible (C57BL/6) mouse strains. An anti-SRIF monoclonal antibody (MAb-SRIF) was used to passively immunize the mice by intraperitoneal injection. The animals were subsequently orally infected with oocysts of E. vermiformis. Individual fecal samples were collected daily for 21 days to monitor the kinetics of oocyst shedding. The fecal oocyst shedding was significantly higher in the C57BL/6 strain than in the BALB/c strain (P < 0.01). Passive immunization with MAb-SRIF in the C57BL/6 mice significantly reduced the number of oocysts in feces (P < 0.05), when compared to the infected non-immunized mice of the same strain. Infected BALB/c mice showed no difference in oocyst shedding in response to the passive immunoneutralization with MAb-SRIF. In conclusion, passive immunization with MAb-SRIF increased resistance to E. vermiformis-infection in the susceptible C57BL/6 mice, but not in the resistant BALB/c mice. This suggests that SRIF modulates gut immune function in parasitic infection.

Animals↗

Environmental contamination by eggs of Toxocara species.

A study was conducted to determine the level of contamination with Toxocara spp. eggs in parks and playgrounds in several central Illinois communities. A total of 135 composite 50-g soil samples were collected from 23 parks and public places in three cities in central Illinois. Of these soil samples, 22 (16.3%) contained from one to thirteen Toxocara spp. eggs. A total of 40 fecal samples were collected from the same parks. Toxocara spp. eggs were found in two (5%) of the samples.

Animals↗

Isotope ratios of molybdenum determined by thermal ionization mass spectrometry for stable isotope studies of molybdenum metabolism in humans.

Methods were developed to separate and purify Mo from biological samples and to measure isotopic ratios in 1 microgram of Mo. A magnetic sector, thermal ionization mass spectrometer was used with simultaneous collection of five isotopes. Isotopic ratios were corrected for mass fractionation by iterative normalization using the 96/98 ratio. Ion beam intensity was enhanced by using a double-filament configuration, loading samples onto evaporation filaments with silica gel and boric acid. A triple-isotope-dilution approach was used, so the method could be applied to two-tracer studies of Mo metabolism in human subjects. 94Mo was added to samples prior to purification to quantify the total Mo content of samples and to determine the amounts of enriched 97Mo and 100Mo appearing in urine and fecal samples of study participants. The three ratios, 94/98, 97/98, and 100/98, were determined with within-run precision of from 0.06 to 0.10% (RSD). Precision of the ratios between replicates was from 0.05 to 0.08%.

Boric Acids↗

The effect of dietary citrus pectin on the excretion of human fecal neutral and acid steroids and the activity of 7alpha-dehydroxylase and beta-glucuronidase.

The purpose of this study was to investigate the effects of citrus pectin on human fecal neutral and acid steroid excretion and beta-glucuronidase and 7alpha-dehydroxylase activity. Eight healthy male subjects (age 20 to 27 yr) were used in a switchback design with or without 15 g citrus pectin added to a mixed low fiber diet. There were three successive 18-day periods preceded by a 4-day adjustment period. Half of the subjects followed a pectin-nonpectin-pectin protocol and the other half followed a nonpectin-pectin-nonpectin protocol. Fecal samples were collected throughout the study under anaerobic conditions. Compared to the control diet, mean fecal weight, percentage moisture, transit time, and fecal fat for both groups of subjects were not significantly different by analysis of variance when subjects were fed pectin diet. Mean beta-glucuronidase activity was increased (35%) when subjects were fed the pectin. Mean 7alpha-dehydroxylase activity showed no definite trend. Mean neutral steroid concentration was slightly decreased (8%) when the pectin diet was fed but total excretion was unchanged. End of period neutral steroid concentration was decreased 9% and total excretion was decreased 3.5%. Mean acid steroid concentration was not changed but total excretion was increased (11%) on the pectin diet. End of period acid steroid concentration and excretion was increased 6% on the pectin diet. This study shows that there were interrelationships between dietary pectin, neutral and acid steroid metabolism, and bacterial enzyme activity.

Adult↗

Dose response for infection by Escherichia coli O157:H7 from outbreak data.

In 1996, an outbreak of E. coli O157:H7-associated illness occurred in an elementary school in Japan. This outbreak has been studied in unusual detail, making this an important case for quantitative risk assessment. The availability of stored samples of the contaminated food allowed reliable estimation of exposure to the pathogens. Collection of fecal samples allowed assessment of the numbers infected, including asymptomatic cases. Comparison to other published dose-response studies for E. coli O157:H7 show that the strain that caused the outbreak studied here must have been considerably more infectious. We use this well-documented incident as an example to demonstrate how such information on the response to a single dose can be used for dose-response assessment. In particular, we demonstrate how the high infectivity limits the uncertainty in the low-dose region.

Adult↗

Periodontal bone level and gingival proteinase activity in gnotobiotic rats immunized with Bacteroides gingivalis.

Bacteroides gingivalis is associated with various forms of periodontal disease. To assess the role of the immune response in modulating B. gingivalis-associated periodontal disease, the effect of immunization of B. gingivalis-induced periodontal bone loss was evaluated in gnotobiotic rats. Male Sprague-Dawley rats immunized with various doses of whole cells or sham-immunized with incomplete Freund's adjuvant were monoinfected with B. gingivalis in carboxymethylcellulose by gavage. Two additional groups served as either sham-immunized or untreated germ-free controls. Forty-two days after infection, all rats were killed, periodontal bone level was assessed morphometrically and radiographically, and gingival proteinase (mammalian collagenase and acid cathepsin) activity was assessed biochemically. B. gingivalis was present in oral samples from all monoinfected rats, and no contaminating bacteria were detected in any oral or fecal sample. Animals immunized with B. gingivalis cells had elevated serum and saliva antibodies to whole cells and partially purified fimbriae from B. gingivalis. Infected sham-immunized rats had significantly more periodontal bone loss than noninfected controls, whereas the periodontal bone level in infected rats immunized with 10(10) B. gingivalis cells was similar to that of the noninfected controls. The activities of gingival collagenase and cathepsin B and L were high in sham-immunized infected rats and low in all other animal groups. In conclusion, it is possible to reduce B. gingivalis-induced periodontal tissue loss in gnotobiotic rats by immunization.

Alveolar Bone Loss↗

Genomic analysis of vaccine-derived poliovirus strains in stool specimens by combination of full-length PCR and oligonucleotide microarray hybridization.

Sabin strains of poliovirus used in the manufacture of oral poliovirus vaccine (OPV) are prone to genetic variations that occur during growth in cell cultures and the organisms of vaccine recipients. Such derivative viruses often have increased neurovirulence and transmissibility, and in some cases they can reestablish chains of transmission in human populations. Monitoring for vaccine-derived polioviruses is an important part of the worldwide campaign to eradicate poliomyelitis. Analysis of vaccine-derived polioviruses requires, as a first step, their isolation in cell cultures, which takes significant time and may yield viral stocks that are not fully representative of the strains present in the original sample. Here we demonstrate that full-length viral cDNA can be PCR amplified directly from stool samples and immediately subjected to genomic analysis by oligonucleotide microarray hybridization and nucleotide sequencing. Most fecal samples from healthy children who received OPV were found to contain variants of Sabin vaccine viruses. Sequence changes in the 5' untranslated region were common, as were changes in the VP1-coding region, including changes in a major antigenic site. Analysis of stool samples taken from cases of acute flaccid paralysis revealed the presence of mixtures of recombinant polioviruses, in addition to the emergence of new sequence variants. Avoiding the need for cell culture isolation dramatically shortened the time needed for identification and analysis of vaccine-derived polioviruses and could be useful for preliminary screening of clinical samples. The amplified full-length viral cDNA can be archived and used to recover live virus for further virological studies.

DNA, Complementary↗

Rat enterohepatic circulation and intestinal distribution of enterally infused thyroid hormones.

The enterohepatic circulation (recycling), intestinal (gut) distribution, metabolism, and excretion of enterally infused thyroid hormones were studied in the intact rat under approximately normal physiological steady state conditions. Rats with 7-day osmotic minipumps implanted ip received constant intraduodenal infusions to steady state of very small trace doses of either 125I-labeled T3 (T3*) or T4 (T4*). Enterohepatic and other pathways remained open to normal function, and in particular, there was no biliary diversion or ligation. Complete feces and urine were collected daily, to assess daily distributions of radioactivity and establishment of the steady state, which occurred by day 3. On day 7, rats were anesthetized, blood was sampled, whole intestine and minipumps were removed, and the gut was separated into six segments. Fecal samples and the contents of each gut section were homogenized, ethanol extracted, evaporated, and reconstituted in NaOH for quantitative aqueous chromatography along with infusate, urine, and plasma samples, on Sephadex G-25 columns. No T3* or T4* was found in urine, but feces contained 39% of the T3* infused and 36% of the T4* infused in steady state. Statistically significant amounts of both T3* and T4* in systemic plasma on day 7 clearly indicated absorption of the hormones from the intestine, distinctly demonstrating an enterohepatic circulation of T3 and T4 under experimental conditions closely approximating the physiological steady state. This also establishes the intestine (with its contents) as an exchangeable hormone pool, physiologically internal to the system regulating thyroid hormones and their distribution. Gut contents contained 52 times more T3* and 4.34 times more T4* than corresponding plasma pools in steady state. Kinetic analysis of the data indicated that somewhat more than half of the T3* or T4* infused was absorbed from gut to liver (primary absorption), and up to 34% of the T3* infused and 43% of the T4* infused reached the systemic circulation (secondary absorption/bioavailability). Gut contents longitudinal distribution data 1) confirm existing evidence that thyroid hormone conjugates, formed elsewhere, exist in gut contents and are hydrolyzed there; 2) demonstrate that deconjugation becomes quantitatively significant, and thus may be initiated, at the level of the cecum; and 3) strongly suggest that absorption of unconjugated hormone occurs from at least the small intestine, all under normal physiological conditions.

Animals↗

Effect of dietary copper source (cupric citrate and cupric sulfate) and concentration on growth performance and fecal copper excretion in weanling pigs.

In each of two experiments, 924 pigs (4.99 kg BW; 16 to 18 d of age) were assigned to 1 of 42 pens based on BW and gender. Pens were allotted randomly to dietary copper (Cu) treatments that consisted of control (10 ppm Cu as cupric sulfate, CuSO4 x 5H2O) and supplemental dietary Cu concentrations of 15, 31, 62, or 125 ppm as cupric citrate (CuCit), or 62 (Exp. 2 only), 125 (Exp. 1 only), or 250 ppm as CuSO4. Live animal performance was determined at the end of the 45-d nursery phase in each experiment. On d 40 of Exp. 2, blood and fecal samples were collected from two randomly selected pigs per pen for evaluation of plasma and fecal Cu concentrations and fecal odor characteristics. In Exp. 1, ADG, ADFI, and G:F were increased (P < 0.05), relative to controls, when pigs were fed diets containing 250 ppm Cu as CuSO4. Pigs fed diets containing 125 ppm Cu as CuCit had increased (P < 0.05) ADG compared with pigs fed diets supplemented with 15 or 62 ppm Cu as CuCit. The ADG, ADFI, and G:F did not differ among pigs fed diets containing 125 and 250 ppm Cu as CuSO4 or 125 ppm Cu as CuCit. In Exp. 2, pigs fed diets containing 250 ppm Cu as CuSO4 had improved (P < 0.05) ADG, ADFI, and G:F compared with controls. In addition, ADG, ADFI, and G:F were similar when pigs were fed diets containing either 250 ppm Cu as CuSO4 or 125 ppm Cu as CuCit. Pigs fed diets containing 62 ppm Cu as CuSO4 or CuCit had similar ADG, ADFI, and G:F. Plasma Cu concentrations were not affected by dietary Cu source or concentration, but fecal Cu concentrations were increased (P < 0.05) as the dietary concentration of Cu increased. Pigs consuming diets supplemented with 125 ppm Cu as CuCit had fecal Cu concentrations that were lower (P < 0.05) than pigs consuming diets supplemented with 250 ppm Cu as CuSO4. Fecal Cu did not differ in pigs receiving diets supplemented with 62 ppm Cu as CuSO4 or CuCit. Odor characteristics of feces were not affected by Cu supplementation or source. These data indicate that 125 and 250 ppm Cu gave similar responses in growth, and that CuCit and CuSO4 were equally effective at stimulating growth and improving G:F in weanling pigs. Fecal Cu excretion was decreased when 125 ppm Cu as CuCit was fed compared with 250 ppm Cu as CuSO4. Therefore, 125 ppm of dietary Cu, regardless of source, may provide an effective environmental alternative to 250 ppm Cu as CuSO4 in weanling pigs.

Animal Feed↗

Persistent efficacy of moxidectin canine sustained-release injectable against experimental infections of Ancylostoma caninum and Uncinaria stenocephala in dogs.

The efficacy of moxidectin canine sustained-release injectable administered at fixed intervals before administration of an oral hookworm challenge was evaluated in 40 laboratory dogs. Groups of eight dogs were treated with the moxidectin sustained-release formulation by SC injection approximately 3, 4, 5, or 6 months before being given oral inoculations with 300 Ancylostoma caninum larvae on Day 0 and 400 Uncinaria stenocephala larvae one day later. Dogs were euthanized 21 days after parasite inoculations. Fecal samples and the intestinal contents collected at necropsy from each dog were examined for hookworms. Fecal egg count reductions based on geometric means relative to controls were 99.2% (3 months) to 81.2% (6 months). The reduction in A. caninum worm recoveries at 3, 4, 5, and 6 months based on geometric means were 94.7%, 90.3%, 82.0%, and 60.2%, respectively. The mean reductions in U. stenocephala worm counts at these intervals were 94.6%, 85.3%, 71.6%, and 48.2%,respectively.

Ancylostoma↗

Fecundity of gastrointestinal trichostrongylid nematodes of sheep in the field.

Twenty-two Dorset Rambouillet lambs were moved to contaminated pasture on Apr 1, 1987. At regular intervals thereafter, pairs of lambs were withdrawn and euthanatized. Gastrointestinal parasites in the abomasum, small intestine, cecum, and large intestine were removed and counted. The last pair of lambs was euthanatized 8 months after original placement on the contaminated pasture. Fecal samples were taken at 3- to 4-week intervals throughout the grazing season and the fecal egg counts were used to estimate parasite fecundity (output of eggs per female parasite per day). The principal parasite genera found included Haemonchus spp, Trichostrongylus spp, and Nematodirus spp. In each of the genera examined, parasite fecundity remained the same irrespective of the intensity or duration of infection. Estimated average fecundities (eggs/female/day) were as follows: Haemonchus contortus, 6,582; Trichostrongylus spp, 262; Nematodirus spp, 40; and Oesophagostomum venulosum, 11,098.

Abomasum↗

Effect of chronic ingestion of tarbush (Flourensia cernua) on ewe lambs.

Efforts to increase livestock utilization of tarbush are being coupled with studies to examine tarbush toxicity. Thirty-eight (19/treatment) ewe lambs were assigned at birth to receive either tarbush or alfalfa (15%, dry matter basis) in a sorghum-based growing ration. Lambs were pen-fed this diet 60 d pre-weaning and 60 d post-weaning. No differences existed between treatments in feed consumption. In the tarbush group, 1 lamb died of unknown causes at 90 d of age, while 3 lambs died between 115 and 120 d of age. There were no deaths in the alfalfa group. Shortly before death, lambs fed tarbush appeared lethargic, disoriented and anorectic. At 122 d of age, 5 lambs were randomly selected from each group. Feces and jugular blood samples were obtained from each lamb before being euthanized and necropsied the following day. All fecal samples were negative for occult blood. Serum gamma glutamyl-transpeptidase (P < 0.001) and aspartate aminotransferase (P < 0.001) activities and platelet counts (P < 0.05) were elevated in lambs fed tarbush, while serum calcium concentrations tended (P < 0.10) to be greater. Histologic examination revealed diffuse liver apoptosis in lambs fed tarbush. These data indicate tarbush leaves cause liver damage when fed for extended periods of time.

Animals↗

Impact of social environment on female chimpanzee reproductive cycles.

The socio-sexual environment of a female is known to affect ovarian function. Increased male contact can enhance menstrual cycle regularity. Conversely, social deprivation constitutes a form of stress that often alters cyclicity and the secretion of reproductive hormones. The present study was carried out on captive female chimpanzees to examine possible interactions among housing conditions, menstrual cycle length, morphological changes in secondary sexual character expression and endocrine release patterns related to follicular and luteal function. Animals were housed over a period of 2 years either with a male conspecific or singly. Blood samples were collected over three cycles, and anogenital swelling changes registered to define menstrual cycle phases. Fecal sampling techniques were used to monitor cortisol as a measure of stress-load. Male presence seemed to affect female cyclicity. Females housed with a male had shorter and more regular cycles than singly housed females. Prolactin, gonadotropins and estradiol levels were generally higher in paired females during specific cycle phases. Group variation was not always significant. No differences were found in progesterone. Sexually cohabited females tended to have lower fecal cortisol metabolites immediately before and after maximum tumescence. We suggest that the close behavioral, physical and olfactory contact with a male conspecific can act as a sort of zeitgeber to modulate ovarian function by stabilizing the female cycle and, perhaps, enhancing folliculogenesis and ovulation.

Analysis of Variance↗

Antimicrobial resistance in human oral and intestinal anaerobic microfloras.

In the present study we determined the resistance patterns of anaerobic bacteria from human saliva and stool specimens and investigated whether there were significant differences in resistance between outpatients and hospitalized patients, regardless of whether they had received antimicrobial agents. No bacterial strains resistant to ampicillin, piperacillin, cefoxitin, cefuroxime, imipenem, clindamycin, doxycycline, chloramphenicol, or metronidazole were isolated from the saliva samples. However, resistance to ampicillin, cefoxitin, and cefuroxime was found in strains from 70% of the fecal samples (mainly Bacteroides thetaiotaomicron, Clostridium innocuum, and Bacteroides ovatus). Resistance to both ampicillin and cefuroxime was frequently found in 19% of the isolated strains (mainly B. thetaiotaomicron, B. ovatus, and Bacteroides vulgatus). No strains that were resistant to imipenem, chloramphenicol, or metronidazole were found. Hospitalization and/or intake of antimicrobial agents was associated with an increase in the relative number of resistant anaerobic intestinal bacteria. The percentage of resistant anaerobic strains encountered, compared with the total number of anaerobic bacteria in the normal fecal microflora, was between 5.2 and 14.8%, with the lower value associated with the outpatient group. Two-thirds of the resistant strains from this group had a relative frequency of less than 1% of the total anaerobic flora, while one-third of the strains were present at a level of greater than 1%; for the hospitalized patients, two-thirds of the strains were present at a level of greater than 1%, and one-third of the strains were present at a level of less than 1% (P < 0.001). Patients who had received antimicrobial agents for 6 days or more (n=20) had an average of 1.6 resistant anaerobic strains each, while patients treated for 3 to 5 days (n = 30) had a mean number of 0.87 resistant strains each ( P < 0.05).

Adult↗

Pharmacokinetics, metabolism, and routes of excretion of intravenous irofulven in patients with advanced solid tumors.

Irofulven is currently in Phase 2 clinical trials against a wide variety of solid tumors and has demonstrated activity in ovarian, prostate, gastrointestinal, and non-small cell lung cancer. The objectives of this study were to determine its pharmacokinetics and route of excretion and to characterize its metabolites in human plasma and urine samples after a 30-min i.v. infusion at a dose of 0.55 mg/kg in patients with advanced solid tumors. Three patients were administered i.v. 100 microCi of [14C]irofulven over a 30-min infusion on day 1 of cycle 1. Serial blood and plasma samples were drawn at 0 (before irofulven infusion) and up to 144 h after the start of infusion. Urine and fecal samples were collected for up to 144 h after the start of infusion. The mean urinary and fecal excretion of radioactivity up to 144 h were 71.2 and 2.9%, respectively, indicating renal excretion was the major route of elimination of [14C]irofulven. The C(max), AUC(0-infinity), and terminal half-life values for total radioactivity were 1130 ng-Eq/ml, 24,400 ng-Eq . h/ml, and 116.5 h, respectively, and the corresponding values for irofulven were 82.7 ng/ml, 65.5 ng . h/ml, and 0.3 h, respectively, suggesting that the total radioactivity in human plasma was a result of the metabolites. Twelve metabolites of irofulven were detected in human urine and plasma by electrospray ionization/tandem mass spectrometry. Among these metabolites, the cyclopropane ring-opened metabolite (M2) of irofulven was found, and seven others were proposed as glucuronide and glutathione conjugates.

Antineoplastic Agents, Alkylating↗

Detection of alkaline sphingomyelinase activity in human stool: proposed role as a new diagnostic and prognostic marker of colorectal cancer.

OBJECTIVES: Intestinal alkaline sphingomyelinase, by exerting a major role in dietary sphingomyelin digestion, is responsible for the generation of messengers able to trigger the rapid turnover and apoptosis in intestinal epithelial cells. Markedly reduced mucosal alkaline sphingomyelinase activity has been associated with human colorectal neoplasms. The aim of this study was to analyze the alkaline sphingomyelinase activity in feces from healthy subjects and colorectal adenocarcinoma patients and to correlate it with the enzyme activity in intestinal tissues. MATERIALS AND METHODS: The enzyme activity was measured both in the intestinal samples from 12 healthy controls and 51 patients with colorectal adenocarcinoma (tumoral and paratumoral tissue) and in the fecal samples of 34 healthy subjects and 29 patients with adenocarcinoma. The relation between sphingomyelinase activity and Dukes' stage, cell differentiation degree, age, and gender was also analyzed. RESULTS: Alkaline sphingomyelinase was significantly decreased (P < 0.001; mean reduction >90%) in tumoral intestinal mucosa of patients compared with controls independently of Dukes' stage and tumor differentiation grade. Interestingly, the enzyme activity in histologically normal paratumoral tissues was statistically lower than control samples (P < 0.001). As occurs in neoplastic tissues, a relevant mean reduction (P < 0.0001; almost 90%) of alkaline sphingomyelinase was revealed in stool samples from tumor patients when compared with controls. CONCLUSION: These findings may have implications for cancer biology and perhaps also for the design of clinical test, thus suggesting that the fecal sphingomyelinase activity could really reflect the human intestinal mucosa enzyme level and could represent a new marker for human colorectal adenocarcinoma, mainly taking into account its early appearance in intestinal neoplasms.

Adenocarcinoma↗

Discrepancies in viral gastroenteritis diagnosis: an unusual dual reovirus-adenovirus infection case.

BACKGROUND: A variable rate of false-positive results may be observed with commercial assays for the detection of rotavirus and adenovirus antigen in stool specimens, depending on the quality of the reagents and the presence of potentially interfering substances in stool samples. OBJECTIVE: The present report analyse the discrepant results that could be obtained by the commercially available diagnostic tests and that can mask the reliable viral diagnosis. STUDY DESIGN: One fecal sample was collected from a hospitalized child aged 6 months with acute watery diarrhea and dehydration. The fecal specimen was processed the same day for the rotavirus and adenovirus antigen detection. RESULTS: The sample was positive for rotavirus antigen by one-step membrane test based on immunochromatographic assays (ICA) and enzyme immunoassays (EIA) monoclonal test but it was negative by an EIA polyclonal test, polyacrylamide gel electrophoresis (PAGE) and RT-PCR assays. In the other hand, the sample was positive for adenovirus antigen by ICA and EIA adenovirus type 40/41. Finally, the sample showed by PAGE an electrophoretic profile resembled that of reovirus. CONCLUSION: The use of a wide repertory of diagnosis tests allowed to reach an unusual reovirus-adenovirus type 40/41 dual infection. This case also point out the potential participation of reovirus in the ethiology of the diarrhea illness.

Adenoviridae Infections↗