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At least 1,153 records · Page 64Linked to original sources

DNA profiling of orangutan (Pongo pygmaeus) feces to prove descent and identity in wildlife animals.

In this study, orangutan (Pongo pygmaeus ssp.) DNA was extracted from excrements, obtained from individual zoo populations. To examine whether human short tandem repeat loci (STR) are suitable for the reconstruction of kinship in orangutans, nine STRs, commonly used in forensic studies in man and the amelogenin system, were amplified in a multiplex polymerase chain reaction (PCR) approach. Five of the nine human autosomal STRs in question amplified successfully in orangutans. Furthermore, the experiments revealed that orangutans possess allele distributions comparable to humans in these systems. Fragment length determination allowed kinship structures to be reconstructed.

Animals↗

Direct flow cytometry of anaerobic bacteria in human feces.

We describe a flow cytometry method for analysis of noncultured anaerobic bacteria present in human fecal suspensions. Nonbacterial fecal compounds, bacterial fragments, and large aggregates could be discriminated from bacteria by staining with propidium iodide (PI) and setting a discriminator on PI fluorescence and by exclusion of events with large forward scatter. Since anaerobic bacteria, which account for over 99.9% of all fecal bacteria, die during sample preparation, a fixation step was not necessary. A second aim of this study was to investigate the technical possibility of measurement of in vivo IgA coating of fecal anaerobic bacteria as well as their bacterial size. Fecal samples of 22 healthy human volunteers were analyzed. The fluorescence distribution of IgA-coated bacteria labeled with fluorescein isothiocyanate (FITC)-anti-Hu-IgA had overlap with noncoated bacteria. However, with match region subtraction, detection of low levels of specific FITC fluorescence on IgA-coated bacteria was achieved. The median bacterial two-dimensional surface area was 1.0 microns2. To validate flow cytometry data, all samples were analyzed with an image analysis system as well. With this new method, a rapid evaluation of fecal flora with high sensitivity for specific FITC fluorescence is possible without culturing.

Adult↗

Chromosome breaking activity of human feces and its enhancement by transition metals.

Chloroform-methanol extracts from human fecal samples were found to contain compounds which induce chromosome aberrations in Chinese hamster ovary cells. The induction of chromosome aberrations is stimulated by Cu2+ or Mn2+ and inhibited by Fe2+ or Fe3+. Addition of catalase to the fecal extract or the mixture of fecal extract and Mn2+ reduced the frequency of chromosome aberrations. These properties are indicative of hydroxyradical producing agents.

Animals↗

Quantitative analyses of beta-carotene and retinol in serum and feces in support of clinical bioavailability studies.

Among more than 50 provitamin carotenoids, beta-carotene is the most metabolically active source of retinol. Despite diets rich in fruits and vegetables containing beta-carotene, vitamin A deficiency is the leading cause of blindness and childhood mortality in developing countries. In addition, the uncertainty of beta-carotene bioconversion into vitamin A suggests that new data are needed to update the nutritional guidelines in developed countries. Previously, we reported the development of a carotene/retinol plateau isotopic enrichment method (CarRet PIE) for the determination of beta-carotene bioavailability and bioconversion into retinol, which utilizes positive ion atmospheric pressure chemical ionization (APCI) liquid chromatography/mass spectrometry (LC/MS). While seeking to validate the CarRet PIE using a mass balance approach requiring fecal measurements of beta-carotene and retinol, interference was encountered that required substantial modifications of the LC/MS assay. Here we report a new LC/MS assay that is based on the detection of molecular anions of beta-carotene using negative ion APCI with a reversed-phase C30 column for HPLC separation. Sample preparation required saponification to eliminate interfering triglycerides. The limit of detection (LOD) of beta-carotene was 0.25 pmol calculated on the basis of an injection of 20 microL of 0.0125 microM beta-carotene, and the limit of quantitation (LOQ) was 1.0 pmol based on the injection of 20 microL of 0.050 microM beta-carotene. The linear range was 1.1 to 2179 pmol on-column. The wide linear range and low LOD and LOQ of this assay facilitated the sensitive and selective quantitative analysis of beta-carotene in both serum and fecal samples in support of an on-going clinical investigation of beta-carotene bioavailability and bioconversion into vitamin A.

Biological Availability↗

Sequence analysis of pigeon, turkey, and chicken rotavirus VP8* identifies rotavirus 993/83, isolated from calf feces, as a pigeon rotavirus.

Partial-length gene 4 cDNA encompassing the VP8* portion of VP4 from chicken rotavirus (RV) Ch-1, turkey RV Ty-1 and Ty-3, and pigeon RV PO-13 was sequenced and compared to RV 993/83 isolated from a calf with diarrhea. Ninety-six percent amino acid sequence identity was seen between VP8* from calf RV 993/83 and pigeon RV PO-13, while only 77 to 83% identity was seen in comparison with turkey and chicken RV VP8* sequences. Phylogenetic tree analysis places all avian RV (including calf isolate 993/83) on a branch at the bottom of the VP8* tree. When tested with three neutralizing monoclonal antibodies raised against pigeon RV PO-13, RV 993/83 and Ty-1 share three, Ch-1 shares two, and Ty-3 shares one neutralizing epitope(s) on VP4 with pigeon RV PO-13.

Amino Acid Sequence↗

Attempt of Yersinia enterocolitica isolation from human feces in Senegal.

One-hundred-eight stool samples, collected in a fishing village of Senegal from 72 apparently healthy subjects and from 36 patients with gastrointestinal disorders, were examined for the presence of Y. enterocolitica. After 1, 2, 3 weeks of cold enrichment with PBS 1/15M, pH 7.6, plating was performed on MacConkey Agar after use of the alkali method. No Yersinia strains were isolated.

Adolescent↗

Coronavirus-like particles in nonhuman primate feces.

Coronavirus-like particles were detected by electron microscopy in normal and diarrhea stool specimens from baboons, chimpanzees, macaques, and marmosets. There was a high prevalence of particles in both normal and diarrhea stools of older animals, whereas stools from nonweaned animals rarely contained these coronavirus-like particles.

Aging↗

Detection of bovine coronavirus in feces by reversed passive hemagglutination.

A reversed passive hemagglutination (RPHA) method was developed for the detection of bovine coronavirus in fecal specimens. Sheep erythrocytes fixed with glutaraldehyde, and then treated with tannic acid were coated with anti-bovine coronavirus rabbit antibodies purified by affinity chromatography using bovine coronavirus linked to Sepharose 4B. The RPHA test was carried out by a microtiter method. Erythrocytes coated with purified specific antibodies were agglutinated by bovine coronavirus, but not by bovine rotavirus or enterovirus. The reaction was inhibited by antiserum to bovine coronavirus, confirming the specificity of the reaction. The RPHA test detected bovine coronavirus in 13 of 22 fecal specimens (59 per cent), from natural cases of diarrhea, while the positive rates were only 14 per cent (3/22) and 22 per cent (5/22) for immunofluorescent staining of primary cultures of calf kidney cells infected with the specimens, and immune electron microscopy respectively. The advantages of the RPHA method are its simplicity, high sensitivity and rapidity.

Animals↗

Detection of group A rotavirus by reverse transcriptase and polymerase chain reaction in feces from children with acute gastroenteritis.

RT-PCR was employed to detect rotavirus infection in 450 fecal samples from children with acute diarrhoea. It was compared with enzyme-linked immunosorbent assay (ELISA) and polyacrylamide gel electrophoresis (PAGE) for rotavirus detection. A total of 67 samples were found positive by at least one of the three techniques. Of these 67 samples, 51 were positive by all three methods, 6 were positive by ELISA and RT-PCR but negative by PAGE, 3 were positive by PAGE and RT-PCR and negative by ELISA, 3 and 4 samples were exclusively positive by RT-PCR and ELISA respectively. These results indicate that RT-PCR is a sensitive and specific assay for detection of group A rotaviruses in stool samples from cases of acute diarrhoea.

Acute Disease↗