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PubMed2005

[ICP-MS with octopole reaction system applied to isotopic analysis of iron in human fecal samples].

OBJECTIVE: The study was aimed to evaluate the application of the quadrupole inductively coupled plasma mass spectrometer (ICP-MS) with collision/reaction cell octopole reaction system (ORS) to determine the iron stable isotope ratio in human fecal samples. METHODS: In a field trial on human metabolism, 3.0 mg per subject of iron with 57Fe enriched was orally administrated, and feces were collected by fecal monitoring. The isotope ratio of 57Fe to 56Fe in the prepared fecal sample was detected with the ORS-ICP-MS. RESULTS: With the application of ORS-ICP-MS, interferences from the Ar plasma and the calcium-contained polyatomic ions could be eliminated. When the sample solution properly diluted, interferences from the matrix were too little to be noted. The relative stand deviation (RSD) of the long-term and short-term precision was < 0.3 %. From the changes of 57 Fe/56 Fe in the daily samples, a regularity of fecal iron outputs was presented, and the fractional iron absorption could be evaluated. CONCLUSION: The ratio of 57 Fe to 56 Fe in the fecal sample could be accurately determined by the quadrupole ICP-MS with ORS, based on which the dietary iron absorption could be evaluated and the metabolism of iron in the human body could also be traced.

Feces↗
PubMed1989

[The occurrence of Treponema in fecal samples from dogs and cats with and without intestinal diseases].

6 (6.9%) of 87 examined dogs without diarrhoea proved to be carriers of Treponema (1x T. hyodysenteriae, 5x T. innocens), whereas in fecal samples from 62 dogs with enteric symptoms no isolation of Treponema succeeded. 5 fecal samples (3.7%) of cats without signs of diarrhoea were found to contain Treponema (1x T. hyodysenteriae, 4x T. innocens), whereas the fecal samples of 31 cats with diarrhoea didn't show any growth of Treponema by cultural investigations. Due to the results of these investigations the conclusion can be drawn that Treponema belong to the usual bacteria of dogs' and cats' intestines and cannot be suspected to cause diarrhoea in these animals primarily.

Animals↗
PubMed1985

Method for improving the detection of viruses in fecal samples.

A method is described that improved the detection of viruses in fecal samples by electron microscopy. The virus particles were concentrated, and much of the background debris was removed by adsorption of viruses on meat protein added to the fecal sample at a low pH and a low salt concentration. Viruses were eluted by raising the pH and the salt concentration. Further concentration was achieved by acid precipitation and vacuum dialysis.

Adsorption↗
PubMed2004

Detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples: comparison of three polymerase chain reaction-based diagnostic tests with a conventional culture method.

Three commercially available assays, designed to specifically detect the presence of Mycobacterium avium subsp. paratuberculosis (MAP) in fecal samples by IS900-PCR, were compared with a conventional culture method. Fecal samples from 100 dairy cows were tested. Fifty-four (67.5%) of 80 culture-positive samples were positive for an assay that detects MAP DNA by dot spot hybridization of polymerase chain reaction products (kit A), 48 (60%) were positive by an assay using ethidium bromide staining for agar gel visualization of amplification products (kit B), and 49 (61.3%) were positive by an assay in which amplified products are detected by a colorimetric detection system (kit C). Relative sensitivity of all tests increased in proportion to the presence of MAP in fecal samples. Specificity was 100% based on results from 20 culture-negative samples from an MAP-free herd.

Animals↗
PubMedDate not supplied

Antibiotic resistance of Escherichia coli in fecal samples of healthy people in two different areas in an industrialized country.

Fecal samples of 310 healthy persons, from two populations from different areas in the Netherlands, were examined for the presence of Escherichia coli resistant to ampicillin, tetracycline, sulfamethoxazole, trimethoprim and nitrofurantoin. High prevalences of resistance were found in both populations, ranging from 28% for trimethoprim to 89% for ampicillin. The percentages of the fecal samples with a dominantly resistant E. coli flora (> 50% resistance) were distinctly lower, ranging from 1% for nitrofurantoin to 21% for tetracycline. No significant differences in the level of resistance were observed between these two comparable populations in two different areas. The susceptibilities to 11 antimicrobial agents of 456 at random isolated E. coli were determined. The percentages of resistance varied widely: from 80% for chloramphenicol to 9% for nitrofurantoin. Only 19% of the isolates were susceptible to all antibiotics tested and 14% were resistant to more than four of the agents tested. Great differences in resistance rates between the two populations examined were seen for chloramphenicol (80% to 41%) and trimethoprim (16% to 36%). The results of this study underscore the presence of a human reservoir of antibiotic resistant microorganisms.

Adolescent↗
PubMed1988

Nitrogen-to-protein conversion factors in animal feed and fecal samples.

Nitrogen-to-protein conversion factors were calculated for diets and fecal samples from three animal species fed low- or high-fiber diets. The conversion factors based on protein contents determined by amino acid analyses were calculated from total N (Kjeldahl analyses) and recovered amino acid plus NH3-N (amino acid analyses). Conversion factors based on protein contents determined by a fluorescamine assay for amino acids were also calculated from total Kjeldahl-N. The conversion factors based on Kjeldahl-N averaged 5.3 +/- .7 for the diets and 4.0 +/- .5 for the feces in the six animal and diet comparisons when calculated from anhydrous amino acid formula weights. The greatest deviations from the traditional 6.25 conversion factor occurred in the fecal samples of ruminant animals fed a corn-alfalfa meal diet at a maintenance level of intake. In contrast, conversion factors based on amino acid plus NH3-N were quite stable. These factors averaged 5.7 +/- .1 for feeds and 5.5 +/- .1 for feces when calculated from anhydrous amino acid residue weights.

Amino Acids↗
PubMed1995

[Occurrence of Salmonella in fecal samples of dogs and cats in northern Bavaria from 1975 to 1994].

103 (3.45%) of 2985 fecal samples from dogs respectively 39 (1.92%) of 2024 fecal samples from cats bacteriologically examined during the years 1975 to 1994 yielded Salmonellae. The 103 Salmonella strains of dogs belonged to 32 different serovars and in the cats the 39 strains found represented a total of 17 different serovars. The most frequently detected serovar was S. typhimurium including var. copenhagen, with 33% in the dogs and 49% in the cats.

Animals↗
PubMed2002

Bacillus thuringiensis in fecal samples from greenhouse workers after exposure to B. thuringiensis-based pesticides.

In a study of occupational exposure to Bacillus thuringiensis, 20 exposed greenhouse workers were examined for Bacillus cereus-like bacteria in fecal samples and on biomonitoring filters. Bacteria with the following characteristics were isolated from eight individuals: intracellular crystalline inclusions characteristic of B. thuringiensis, genes for and production of B. cereus enterotoxins, and positivity for cry11 as determined by PCR. DNA fingerprints of the fecal isolates were identical to those of strains isolated from the commercial products used. Work processes (i.e., spraying) correlated with the presence of B. thuringiensis in the fecal samples (10(2) to 10(3) CFU/g of feces). However, no gastrointestinal symptoms correlated with the presence of B. thuringiensis in the fecal samples.

Bacillus cereus↗
PubMed1993

Cultivation of Mycobacterium paratuberculosis from bovine fecal samples by using elements of the Roche MB Check system.

Components of a commercially available, nonradiometric, biphasic (liquid medium then solid medium) system for the detection of Mycobacterial species, Roche MB Check, were adapted for the isolation of Mycobacterium paratuberculosis from bovine fecal specimens. A two-stage culture procedure was developed in which processed fecal samples were incubated in modified commercial liquid medium and then subcultured onto Herrold's egg yolk medium with mycobactin. By using known culture-positive samples and/or samples from animals clinically affected with paratuberculosis, it was found that visible colonial growth on solid media could be obtained after 4 weeks of incubation in liquid medium containing egg yolk and mycobactin followed by 8 weeks of incubation on Herrold's egg yolk medium. In the second part of the study, conventional fecal culture (sample sedimentation in hexadecylcetylpyridinium chloride followed by incubation on Herrold's egg yolk medium) was compared with the two-stage system using a two-step centrifugation technique for sample preparation. One hundred fecal samples from clinically normal but absorbed-enzyme immunoassay-positive cattle were used for the comparison. Conventional culture yielded a sensitivity of 16.5%, whereas the sensitivity of the two-stage system was 29.4%. When used in parallel, the tests detected 36.5% of the samples. There was no significant difference between the two methods in the time taken to obtain visible colonies. These results indicate that the two-stage method is a sensitive method for isolation of M. paratuberculosis from fecal samples obtained from cattle with clinical paratuberculosis. In addition, the two-stage system is more sensitive than conventional culture for the isolation of M. paratuberculosis from subclinically infected cattle.

Animals↗
PubMed1993

Detection of toxigenic Clostridium difficile in fecal samples by colony blot hybridization.

A hybridization assay for detection of toxigenic Clostridium difficile in fecal samples was developed and compared with the classical tissue culture cytotoxicity assay. A DNA fragment probe specific for the toxin B gene of Clostridium difficile was synthesized by the polymerase chain reaction and labelled with digoxigenin. Fecal samples were cultured for 24 hours, replica-plated and hybridized with the probe. The hybridization assay had a sensitivity of 100%, specificity of 96.7%, positive predictive value of 86.7% and negative predictive value of 100% compared with the cytotoxicity assay.

Base Sequence↗
PubMed2003

Fecal samples as DNA source for the diagnosis of Necrotizing Hepatopancreatitis (NHP) in Penaeus vannamei broodstock.

Necrotizing Hepatopancreatitis (NHP) is a severe disease of cultivated penaeid shrimp caused by a pleomorphic, gram-negative, intracellular rickettsia-like bacterium. Current diagnostic methods for this disease are invasive, requiring dissection of the animal to perform histopathological analysis. In Colombia, NHP affects mainly broodstock, being a major cause of mortalities in maturation laboratories. In order to identify the presence of NHP without having to dissect the animal, we developed a PCR-based method using fecal samples as the DNA source. The DNA was extracted using a quick isolation protocol followed by amplification with primers specific for 16S ribosomal RNA gene sequences. To verify the sensitivity and specificity we analyzed samples from the same animal by PCR and in situ hybridization, and found 100% agreement. In addition, we amplified DNA extracted form paraffin blocks to confirm NHP diagnosis. PCR amplification from fecal samples and paraffin blocks yielded the expected 440 bp fragment. We conclude that PCR amplification from fecal samples is a valuable tool for the diagnosis of NHP in broodstock organisms, and that paraffin-fixed tissues can be used as a source of DNA for PCR amplification of NHP.

Animals↗
PubMed1991

Assessment of mean oocyst count in groups of kids: litter, individual randomized and non-randomized fecal samplings.

A trial was conducted in six pens of kids visited five times monthly to compare different regimes of fecal sampling in order to assess average coccidial infection at a group level. In addition to individual fecal samplings of all the kids in the pen, giving the actual mean oocyst count, three sampling methods were performed: (i) individual sampling of three, five, seven, ten, 15, 20 or more animals according to the numbers of animals in the pen; (ii) sampling of feces in the litter; (iii) individual sampling of six kids, three with the poorest and three with the highest daily weight gain. Results indicated that drawing a random sample of ten kids was necessary to obtain a good estimation, with an accuracy of 25% in more than 90% of pens. Sampling of feces in the litter and individual sampling of six kids selected on daily weight gain considerations gave accurate estimation of the actual mean oocyst counts except in the case of high levels of infection. Further investigations would be needed to assess the diagnostic value of such sampling techniques whatever the coccidial infection level.

Age Factors↗
PubMed2004

Detection of diarrheagenic viruses from diarrheal fecal samples collected from children in Kathmandu, Nepal.

Diarrhea causing viruses (Rotavirus. Adenovirus and Norovirus) were investigated in diarrheal fecal samples collected from children in Kathmandu, Nepal in Janury 2004 using both real time PCR and immuno-chromatogaphic techniques. Of the total 12 diarrheal samples investigated, 8 (66.7%) were positive for Rotavirus, 1 (8.3%) was positive for Adenovirus and none was positive for Norovirus (Norwalk like virus). The Adenovirus positive sample was also positive for Rotavirus. Similar results were obtained by immuno-chromatographic technique. All of Rotavirus detected belonged to Group-A. Results indicated that immuno-chromatographic technique was equally good in the detection of diarrhea causing viruses in fecal samples. Furthermore, it was simple, cost-effective and less time consuming (15 minutes) compared with the PCR. Immuno-chromatographic technique, therefore, appeared to be useful for rapid diagnosis of viral gastroenteritis in developing countries like Nepal.

Adenovirus Infections, Human↗
PubMed2002

Molecular characterization of Salmonella spp. isolated from bulk tank milk and cull dairy cow fecal samples.

The consumption of meat from cull dairy cows and of raw milk has been associated with foodborne salmonellosis. This survey was conducted to establish the prevalence of Salmonella in cull dairy cow fecal samples and bulk tank milk and to determine the proportion of Salmonella-positive dairy farms (n = 30) in east Tennessee. Food and Drug Administration bacteriological analytical protocols were generally used for Salmonella isolation. Primary enrichment was performed with lactose broth, and secondary enrichment was conducted with tetrathionate broth. Eosin methylene blue, hektoen enteric, xylose lysine desoxycholate, bismuth sulfite, and brilliant green (BG) were used as isolation agars. BG agars supplemented with individual antibiotics and/or sulfur compounds were also evaluated. Six of 268 (2.24%) bulk tank milk samples and 9 of 415 (2.17%) fecal samples from 7 of 30 (25.3%) dairy farms were Salmonella-positive. Most isolates (11 of 15) were obtained between September and December. Salmonella isolates were further characterized using polyvalent somatic O Salmonella antiserum, o-nitrophenyl-beta-D-galactopyranoside (ONPG), and Analytical Profile Index (API) 20E strips for Enterobacteriaceae. Serological evaluation of presumptive positive Salmonella isolates resulted in substantial numbers of false positives (41.2%). ONPG and API 20E tests enabled further biochemical distinction of the majority of Salmonella spp. from Salmonella Arizonae and closely related members of Enterobacteriaceae like Citrobacter youngae. Pulsed-field gel electrophoresis of SpeI-digested Salmonella DNA was used to subtype isolates. The isolates grouped into four clusters. The baseline information generated in this survey is being used to develop preharvest pathogen reduction programs on selected farms.

Animals↗
PubMed2004

DNA microarray analysis of predominant human intestinal bacteria in fecal samples.

A microarray method was developed for the detection of 40 bacterial species reported in the literature to be predominant in the human gastrointestinal tract. The 40 species include seven species each of Bacteroides and Clostridium, six species of Ruminococcus, five species of Bifidobacterium, four species of Eubacterium, two species each of Fusobacterium, Lactobacillus and Enterococcus, and single species each of Collinsella, Eggerthella, Escherichia, Faecalibacterium and Finegoldia. Three 40-mer oligos specific for each bacterial species were designed based on comparison of the 16S rDNA sequences available in the GenBank database, and were used to make the DNA-array on epoxy slides. Using two universal primers, the 16S rRNA gene from bacteria present in fecal samples were amplified and labeled with Cyanine5-dCTP by PCR, and then hybridized to the DNA-array. After resolving some difficulties caused by sequence conflicts in GenBank and inaccurate reference strains, all 40 bacterial reference species gave positive results. The microarray method was used to screen fecal samples obtained from 11 healthy human volunteers for the presence of these intestinal bacteria. The results indicated that 25-37 of the 40 species could be detected in each fecal sample and that 33 of the species were found in a majority of the samples.

Bacteria↗
PubMed2006

Rapid detection of noroviruses in fecal samples and shellfish by nucleic acid sequence-based amplification.

The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (NASBA) method of detecting noroviruses in artificially and naturally contaminated shellfish. We used 58 fecal samples that tested positive for noroviruses with electron microscopy (EM) to develop an NASBA assay for these viruses. Oligonucleotide primers targeting the polymerase coding region were used to amplify the viral RNA in an isothermal process that resulted in the accumulation of RNA amplicons. These amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup I (GI) and genogroup II (GII) of noroviruses. The expected band of 327 bp appeared in denaturing agarose gel without any nonspecific band. The specific signal for each amplicon was obtained through Northern blotting in many repeats. All fecal samples of which 46 (79.3%) belonged to GII and 12 (20.6%) belonged to GI were positive for noroviruses by EM and by NASBA. Target RNA concentrations as low as 5 pg/ml were detected in fecal specimens using NASBA. When the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100 pg/1.5 g shellfish tissue. The potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in Guangzhou city of China, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6%) belonged to GII and 5 (20.4%) belonged to GI. The NASBA assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity.

Animals↗
PubMed1989

[Comparative studies of the detection of rotavirus in fecal samples of calves with diarrhea with the latex test "Slidex Rota-Kit 2" and electron microscopy].

Of 68 fecal samples from calves with diarrhoea which were tested for rotavirus with the latex agglutination test "Slidex Rota-Kit2" and by electron microscopy 33 samples were positive and 33 were negative with both tests respectively. Divergent results (latex test positive/EM negative and vice versa) were observed in one specimen only, respectively. Cross reactions with other viruses diagnosed by electron microscopy were not observed with the latex agglutination test. The "Slidex Rota-Kit2" is another suitable test for the diagnostic laboratory as well as for the veterinary practitioner for the detection of rotavirus in fecal samples of calves.

Animals↗
PubMed2005

Detection of viral, bacterial, and parasitological RNA or DNA of nine intestinal pathogens in fecal samples archived as part of the english infectious intestinal disease study: assessment of the stability of target nucleic acid.

Fecal samples were collected from cases and controls as part of the Infectious Intestinal Disease (IID) study in England and were stored as frozen suspensions for 8 to 12 years. The purpose of this study was to apply PCR-based procedures to assess the stability of pathogen-specific nucleic acid sequences present in this archive. Samples from which Cryptosporidium, Giardia, Salmonella, Campylobacter, enteroaggregative Escherichia coli (EAggEC), enterotoxigenic Clostridium perfringens, rotaviruses, noroviruses, or sapoviruses had been previously detected during the IID study using conventional methods were selected from the archive. A generic nucleic acid extraction method to recover RNA or DNA was used. Complementary DNA was generated from RNA by reverse transcription with random priming. Block-based and real-time PCR assays were used to amplify and detect gene fragments from each of these pathogens. The percentage reconfirmation of target was as follows: Giardia duodenalis 68%, Cryptosporidium 96%, Campylobacter 98%, Salmonella 98%, enterotoxigenic C perfringens 34%, EAggEC 93.3%, rotavirus 95%, norovirus 73%, and sapovirus 85%. This study has shown that nucleic acid can be extracted and specific sequences amplified and detected from archived fecal samples. The IID archive therefore represents a valuable resource for further studies, especially the investigation of the samples from which no pathogens had previously been detected.

Cryopreservation↗