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Pancreatic elastase 1 in feces of preterm and term infants.

BACKGROUND: Determination of fecal pancreatic elastase 1 (E1) is a reliable and noninvasive test of exocrine pancreatic function. Adult reference values of greater than 200 microg E1/g feces do not seem to be applicable to early infancy because of immature pancreatic function. Because reference values for infants do not exist, the current study was aimed to define reference values for preterm and term infants up to 12 months of age. METHODS: The authors measured pancreatic E1 concentration in feces of 148 infants up to 12 months of age. Infants with known bowel or pancreatic disorders were excluded from the study. RESULTS: The authors found that 96.8% of all children had E1 concentrations greater than an adult lower limit after 2 weeks of life, independent of gestational age. Up to 48 hours after birth, none of the preterm infants had an E1 concentration of greater than 30 microg/g meconium, whereas 43% of the term infants had normal adult values. CONCLUSIONS: The adult reference value for pancreatic E1 of greater than 200 microg/g feces can be applied to infants older than 2 weeks, independent of gestational age, birth weight, and the type of nutrition.

Age Factors↗

Separate isolation of Clostridium difficile spores and vegetative cells from the feces of newborn infants.

A modified taurocholate-cefoxitin-cycloserine-fructose agar medium, pH 5.5, on which vegetative cells alone could grow, was newly devised for separate isolation of Clostridium difficile vegetative cells and spores from feces. The ratio of C. difficile-positive feces from healthy newborn infants younger than 10 days of the age was 30.8%, and 93.3% of feces from healthy infants older than 20 days were positive for C. difficile. C. difficile spores alone were detected in twenty-one samples (75%) of C. difficile-positive Twenty-eight specimens. Only 10.7% (3/28) C. difficile vegetative cells alone were detected. C. difficile spores alone were detected in one of nine healthy adults. These collective results offer potential explanations for high frequent isolations of C. difficile from newborn infants without occurrence of pseudomembranous colitis.

Bottle Feeding↗

Evaluation of a new enzyme immunoassay for detecting Helicobacter pylori in feces: a prospective pilot study.

OBJECTIVE: There is an increasing interest in noninvasive tests for detecting Helicobacter pylori (H. pylori) infection. Unlike serological and urea breath tests, the possibility of searching for H. pylori in feces has been scarcely investigated. The aim of this prospective pilot study was to evaluate the usefulness of a new enzyme immunoassay for detecting H. pylori antigens in feces, as a predictor of H. pylori status in the pre- and posttreatment settings. METHODS: One hundred and fifty-four symptomatic, anti-H. pylori untreated patients (Group A) and 116 anti-H. pylori treated patients (Group B) underwent gastroscopy with biopsies of the antrum and corpus for histology (H) and rapid urease test (RUT). In the anti-H. pylori treated group, a 13C-urea breath test (UBT) was also performed. In Group A, H. pylori status was defined as positive or negative when both H and RUT gave concordant positive or negative results. In Group B, the patients were considered eradicated if all three tests were negative. A stool specimen was collected from all patients the day after gastroscopy, and tested by using an enzyme immunoassay commercial kit for detecting H. pylori antigens in feces (HpSAT). RESULTS: Eighty-five patients in Group A (55%) and 44 in Group B (38%) were H. pylori infected. On the whole, HpSAT showed a sensitivity of 94% and specificity of 86%. In Group A and Group B, sensitivity and specificity were 94% versus 93%, and 90% versus 82%, respectively (p < 0.05). CONCLUSIONS: HpSAT seems to be a reliable method for predicting H. pylori status in anti-H. pylori untreated patients. Conversely, the test appears less suitable to evaluate the outcome of the eradicating treatment. Consequently, it is likely to be accepted for the primary diagnosis of H. pylori status, particularly in dyspeptic young patients.

Adult↗

[A healthy carrier of verotoxin-producing Escherichia coli, who was detected by periodic feces examinations].

Verotoxin-producing Escherichia coli (VTEC) was rapidly detected by the PCR method in one of the 9 feces samples. They were collected from the people who had been cooking meals for patients on a periodic feces examination on November 25, 1992. It was confirmed by PCR that the isolate had both VT1 and VT2vh toxic genes and showed cytotoxicity on Verocells. In this case, the isolate was highly susceptible to common antibiotic agents, it was removed by the administration of tosfulaxacin. Some isolates which had the same properties as those of the strain described in "VITEK GNI card" and the antimicrobial susceptibility and VT toxin types, were detected in 2 of the 3 members of her family, they were healthy carriers and did not have any subjective symptoms such as diarrhea or abdominal pain. In order to detect Verotoxic genes, the sample of feces preincubated for 3 h in trypticase broth was subjected to PCR. We recommend that this method is much more useful because of rapid detection and identification of VTEC compared with the classical culture method.

Adult↗

[Detection of extended spectrum beta-lactamases producing Enterobacteriaceae in feces].

A study was made of 366 feces for detection of extended spectrum beta-lactamases producing Enterobacteriaceae from feces. The selective agar was used for modified drigalski agar (Eiken Chemical Co., LTD) with 2 micrograms/ml cefotaxime (ESBL screen agar). 92 strains of Enterobacteriaceae, 41 Escherichia coli, 15 Citrobacter freundii, 13 Enterobacter cloacae, 11 Klebsiella pneumoniae, and other 12, were isolated from ESBL screen agar. And, R-plasmid that were selected by 2 micrograms/ml cefotaxime were transferred by conjugation from two of the 92 strains. These strain were E. coli TH9809927 and Proteus mirabilis TH9808262 that were amplified by "Toho-1 type" primer. The clude enzyme from two strains (donor) and transconjugants were especially hydrolysed cepharoridine and cefotaxime. Accordingly, two strains (0.5%) were detected as ESBL producers. We think that the result of our survey suggests the increase of ESBLs producing bacterial infection in Japan, and believe that there is a trend of infection of its by surveilance of the feces.

Bacteriological Techniques↗

Presence of Clostridium difficile and antibiotic and beta-lactamase activities in feces of volunteers treated with oral cefixime, oral cefpodoxime proxetil, or placebo.

Three groups of six healthy adult volunteers were randomly assigned to a treatment with 400 mg of oral cefpodoxime proxetil, oral cefixime, or placebo per day for 10 days. Informed consent was obtained from all volunteers. Clostridium difficile was not detected in the feces of any subject before treatment or at any time in the subjects in the placebo group. C. difficile was, however, detected in all subjects treated with cefpodoxime proxetil and in five of six treated with cefixime. Genomic DNA restriction patterns showed that the strains of C. difficile differed from one volunteer to another. Two subjects both shed different strains at different times during the 25-day surveillance period. All isolates were resistant to cefixime and cefpodoxime (MIC for 90% of strains, 256 and 512 mg/liter, respectively). Antibiotic activity was found in the feces of one volunteer treated with cefpodoxime proxetil and of four volunteers treated with cefixime. It was inversely correlated with the presence of fecal beta-lactamase activity. Intestinal side effects were limited to modifications of stool consistency, which occurred in only 3 of the 12 treated volunteers and did not lead to cessation of treatment. These modifications were significantly associated with the presence of fecal antibiotic activity (P less than 0.05) but not with the shedding of toxigenic or nontoxigenic strains of C. difficile or with the presence of toxin A in feces, which was detected only in one perfectly healthy treated volunteer.

Administration, Oral↗

Binding of aminoglycosides to feces.

Gentamicin and several other basic antibiotics were examined for their ability to adsorb to dog feces. It was found that 44 to 90% of all antibiotics studied were adsorbed to feces depending on the ratio of antibiotic to fecal material. Attempts to extract these antibiotics by acid treatment after adsorption onto feces were only partially successful since large portions of the bound materials were not removed.

Adsorption↗

Studies on bacteriophage distribution: virulent and temperate bacteriophage content of mammalian feces.

Freshly voided samples of the feces of cows, pigs, and humans were analyzed for the enumeration of cell-free plaque-forming units (PFU) of coliphages and Salmonella phages. Coliphage PFU counts per gram (wet weight) of feces were found to range from less than 10(1) to greater than 10(7). Salmonella phages were found in three out of five porcine samples, but none were found in the four bovine samples analyzed. Virulent coliphages related to the phiX174/S13 serological group showed some "habitat preference" in that the S13 type of phages was found only in pig feces, whereas the phiX174 type of phages was found only in cow dung. Temperate coliphages were detectable in a majority of samples of both human and porcine origin but were infrequently found in bovine samples. About 80% of the temperate coliphages of fecal origin have been found to be serologically related to phage HK022 (Dhillon and Dhillon, 1973), and all are efficiently inducible by ultraviolet light irradiation. However, considerable diversity with the group was found when the prophage immunity pattern of 10 randomly selected isolates was examined.

Animals↗

Survival of two enterobacteria in feces buried in soil under field conditions.

Feces samples, inoculated with 10(6) Escherichia coli resistant to streptomycin and nalidixic acid and with 10(5) Salmonella typhimurium per g, were buried at five mountain field sites ranging from 2,005 to 2,730 m in elevation. Counts of each bacterium rose initially and then declined to 10(3) or 10(4) per g of feces in 8 weeks. The survival pattern was similar at all sites regardless of marked differences in elevation, soil, moisture, exposure, and vegetation. S. typhimurium numbers were consistently higher than E. coli numbers after week 3. The test encompassed most of the time that the area is snow-free and accessible for hiking. The results were judged to discredit the recommendation for shallow burial of feces and to indicate a potential health hazard under intensive use.

Escherichia coli↗

Immunofluorescent evidence of Proteus mirabilis swarm cell formation on sterilized rat feces.

Swarming Proteus spp. were detected with the use of proteometry (a most-probable-number technique) in the fecal material of selected animal species and in raw sewage from a local sewage treatment plant. Proteus spp. were not detected in any of several soil and freshwater samples examined. Since rat feces harbored high numbers of Proteus mirabilis compared with other habitats examined, we chose to examine it for the possibility of supporting swarming. Immunofluorescent studies with a strain-specific conjugate revealed the morphogenesis of short forms into elongated swarm cells upon the surface of sterilized rat feces that had been inoculated with short forms of P. mirabilis. the same phenomenon was not observed consistently when nonsterile rat feces were inoculated and examined with immunofluorescence.

Animals↗

Occurrence of Rhodococcus coprophilus and associated actinomycetes in feces, sewage, and freshwater.

Freshwater, sewage, and fecal samples from various sources were examined for Rhodococcus coprophilus, associated actinomycetes, Escherichia coli, and fecal streptococci. Rhodococcus coprophilus was isolated consistently from feces of farm animals, poultry reared in proximity to farm animals, freshwater, and wastewater polluted with animal fecal wastes. It was not isolated from samples of human feces. The ratio of R. coprophilus total actinomycetes was higher in feces from cattle, sheep, ducks, and geese than in specimens from pigs, horses, and fowl. In samples from two freshwater streams polluted by fecal material from farm animals, the ratios of R. copropilus to total actinomycetes were similar to those found in fecal specimens from cattle and sheep. Ratios of fecal coliform to fecal streptococci could not distinguish between fresh human and animal fecal samples and, furthermore, were not reflected in the stream waters polluted by animal fecal material. R. coprophilus has potential in water and dairy bacteriology as a specific indicator organism of fecal pollution due to farm animal wastes.

Actinomycetaceae↗

Use of a species-specific DNA hybridization probe for enumerating Bacteroides vulgatus in human feces.

pBV-1, a recombinant plasmid that contains a chromosomal DNA fragment from Bacteroides vulgatus, hybridized to DNA from B. vulgatus but not to DNA from other colonic Bacteroides species. This plasmid was used as a DNA probe to detect and enumerate B. vulgatus in pure culture, in mixed cultures, and in a bacterial fraction from human feces. Bacteria in a pure or mixed culture were lysed by heating the culture in NaOH. The DNA in the disrupted cell suspension was then trapped on nitrocellulose paper by vacuum filtration. If fecal samples were used instead of pure or mixed cultures, it was first necessary to partially purify the DNA by low-speed centrifugation (2,000 X g) and phenol-chloroform extraction before filtering. When 32P-labeled pBV-1 was incubated with filters containg B. vulgatus DNA, the amount of radioactivity that bound to the filters was proportional to the number of B. vulgatus filtered as long as the filtering capacity of the nitrocellulose was not exceeded. Using this procedure, we obtained a value for the concentration of B. vulgatus in human feces (2 X 10(10) to 3 X 10(10) per g of dry weight) that is similar to values obtained by other investigators using conventional bacteriological techniques (3 X 10(10) to 6 X 10(10) per g of dry weight). The advantage of the DNA hybridization method over conventional techniques is that it is not necessary to isolate pure cultures of bacteria from complex specimens such as feces. Furthermore, our method bypasses the cumbersome set of biochemical tests normally used to identify anaerobic bacteria. The major limitation of our method is its sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteroides↗

Monitoring of Yersinia enterocolitica in murine and bovine feces on the basis of the chromosomally integrated luxAB marker gene.

We previously integrated the luxAB gene into the Yersinia enterocolitica chromosome. In this article, we assessed, by luminometry, the survival of the engineered strain KNG1024 in the digestive tracts of mice and cows. In situ detection and a count of the released strain were performed on feces from orally inoculated BALB/c mice for 24 days. This method is a rapid and reliable system for long-term monitoring of genetically engineered bacteria. In cow feces, the count of Y. enterocolitica ranged from 210 to 6,000 CFU/g of feces. This very low count was not detectable by direct luminometry.

Animals↗

Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rRNA-targeted oligonucleotide probes.

Six 16S rRNA-targeted oligonucleotide probes were designed, validated, and used to quantify predominant groups of anaerobic bacteria in human fecal samples. A set of two probes was specific for species of the Bacteroides fragilis group and the species Bacteroides distasonis. Two others were designed to detect species of the Clostridium histolyticum and the Clostridium lituseburense groups. Another probe was designed for the genera Streptococcus and Lactococcus, and the final probe was designed for the species of the Clostridium coccoides-Eubacterium rectale group. The temperature of dissociation of each of the probes was determined. The specificities of the probes for a collection of target and reference organisms were tested by dot blot hybridization and fluorescent in situ hybridization (FISH). The new probes were used in initial FISH experiments to enumerate human fecal bacteria. The combination of the two Bacteroides-specific probes detected a mean of 5.4 x 10(10) cells per g (dry weight) of feces; the Clostridium coccoides-Eubacterium rectale group-specific probe detected a mean of 7.2 x 10(10) cells per g (dry weight) of feces. The Clostridium histolyticum, Clostridium lituseburense, and Streptococcus-Lactococcus group-specific probes detected only numbers of cells ranging from 1 x 10(7) to 7 x 10(8) per g (dry weight) of feces. Three of the newly designed probes and three additional probes were used in further FISH experiments to study the fecal flora composition of nine volunteers over a period of 8 months. The combination of probes was able to detect at least two-thirds of the fecal flora. The normal biological variations within the fecal populations of the volunteers were determined and indicated that these variations should be considered when evaluating the effects of agents modulating the flora.

Adult↗

Development of 16S rRNA-based probes for the Coriobacterium group and the Atopobium cluster and their application for enumeration of Coriobacteriaceae in human feces from volunteers of different age groups.

Two 16S rRNA-targeted probes were developed: one for the Coriobacterium group and the other for the Atopobium cluster (which comprises most of the Coriobacteriaceae species, including the Coriobacterium group). The new probes were based on sequences of three new Coriobacteriaceae strains isolated from human feces and clinical material and sequences from databases. Application of the probes to fecal samples showed that formula-fed infants had higher numbers of Coriobacterium group cells in their feces than breast-fed infants. In addition, based on the presented results, it is hypothesized that with the increasing age of a person, the diversity of Atopobium cluster species present in the feces increases.

Actinobacteria↗

Effect of raw-milk cheese consumption on the enterococcal flora of human feces.

Enterococci are one of the major facultative anaerobic bacterial groups that reside in the human gastrointestinal tract. In the present study, the composition of the enterococcal fecal flora in three healthy humans was analyzed before, during, and after the daily consumption of approximately 125 g of a raw-milk Cheddar-type cheese containing 3.2 x 10(4) enterococci/g of cheese. Enterococcal counts ranged between 1.4 x 10(2) and 2.5 x 10(8) CFU/g of feces and differed from subject to subject and from week to week. The cheese contained mainly Enterococcus casseliflavus and a small population of Enterococcus faecalis. Clonal relationships were determined by pulsed-field gel electrophoresis. Before and after consumption of the cheese, samples from humans contained mainly Enterococcus faecium, with some of the clones being resident. During consumption of the cheese, one particular transient clone of E. faecalis, clone Fs2, which was present in small numbers in the cheese, largely dominated the feces. Two clones of E. casseliflavus from the cheese were also found in the feces of one of the subjects during cheese consumption. These results suggest that a clone need not be present in a food in high numbers to establish itself in the intestine.

Animals↗

Use of pulsed-field gel electrophoresis to characterize the heterogeneity and clonality of salmonella isolates obtained from the carcasses and feces of swine at slaughter.

Salmonella enterica isolates were recovered from swine at a collaborating processing plant over a 2-month period in the spring of 2000. In the present study, molecular subtyping by pulsed-field gel electrophoresis (PFGE) was performed on the 581 confirmed Salmonella isolates from the 84 Salmonella-positive samples obtained from the previous study. A total of 32 different PFGE pulsotypes were observed visually, and a BioNumerics software analysis clustered those pulsotypes into 12 PFGE groups. The B, F, and G groups predominated throughout the sampling period and were isolated from 39, 22, and 13% of the swine, respectively. In addition, multiple isolates were obtained from 67 of the 84 Salmonella-positive samples, and subtyping revealed multiple PFGE profiles in 35 of these 67 (62%) samples. Both carcass and fecal isolates of Salmonella were recovered from 13 swine, resulting in "matched" samples. Molecular typing of the 252 isolates recovered from the matched samples revealed that 7 (54%) of the 13 carcasses were contaminated with Salmonella pulsotypes that were not isolated from the feces of the same animal. Conversely, from 6 of the 13 (46%) matched animals, Salmonella clonal types were isolated from the feces that were not isolated from the carcass of the same animal. These data establish that each lot of swine introduces new contaminants into the plant environment and that swine feces from one animal can contaminate many carcasses. In addition, these results indicate that the examination of multiple Salmonella isolates from positive samples is necessary to determine the variety of potential contaminants of swine carcasses during slaughter and processing.

Abattoirs↗

Concentrations of pathogens and indicators in animal feces in the Sydney watershed.

A fecal analysis survey was undertaken to quantify animal inputs of pathogenic and indicator microorganisms in the temperate watersheds of Sydney, Australia. The feces from a range of domestic animals and wildlife were analyzed for the indicator bacteria fecal coliforms and Clostridium perfringens spores, the pathogenic protozoa Cryptosporidium and Giardia, and the enteric viruses adenovirus, enterovirus, and reovirus. Pathogen and fecal indicator concentrations were generally higher in domestic animal feces than in wildlife feces. Future studies to quantify potential pathogen risks in drinking-water watersheds should thus focus on quantifying pathogen loads from domestic animals and livestock rather than wildlife.

Animals↗