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At least 127 records · Page 7Linked to original sources
PubMed2006

PCR amplification of the IS6110 insertion element of Mycobacterium tuberculosis in fecal samples from patients with intestinal tuberculosis.

PCR amplification of insertion element IS6110 of Mycobacterium tuberculosis in fecal samples was evaluated in the diagnosis of intestinal tuberculosis (ITB). The numbers of samples that tested positive by PCR with SalI digestion were 16/18 untreated-ITB samples, 0/8 treated-ITB samples, 12/14 smear-positive pulmonary tuberculosis samples, and 0/30 control samples. The sensitivity, specificity, positive predictive value, and negative predictive value of fecal PCR were 88.8%, 100%, 100%, and 93.7%, respectively.

Base Sequence↗
PubMed2004

Improved extraction of PCR-quality community DNA from digesta and fecal samples.

Several DNA extraction methods have been reported for use with digesta or fecal samples, but problems are often encountered in terms of relatively low DNA yields and/or recovering DNA free of inhibitory substances. Here we report a modified method to extract PCR-quality microbial community DNA from these types of samples, which employs bead beating in the presence of high concentrations of sodium dodecyl sulfate (SDS), salt, and EDTA, and with subsequent DNA purification by QIAamp columns [referred to as repeated bead beating plus column (RBB + C) method]. The RBB + C method resulted in a 1.5- to 6-fold increase in DNA yield when compared to three other widely used methods. The community DNA prepared with the RBB + C method was also free of inhibitory substances and resulted in improved denaturing gradient gel electrophoresis (DGGE) profiles, which is indicative of a more complete lysis and representation of microbial diversity present in such samples.

Animals↗
PubMed2000

Beta-lactamase patterns and betalactam/clavulanic acid resistance in Escherichia coli isolated from fecal samples from healthy volunteers.

Fecal specimens from 50 healthy volunteers living in Izmir, Turkey, were examined for the presence of beta-lactamase producing Escherichia coli by selection on agar plates containing ampicillin (10 mg/L). Thirty-nine (78%) of the strains were ampicillin-resistant and ampicillin MIC50 values for these isolates were > or =1024 microg/ml (range 32- > or =1024 microg/ml). Ampicillin MIC values remained above 64 microg/ml in 16 (41%) strains despite addition of clavulanic acid (2 mg/L). Beta-lactamase production of the clavulanate-resistant strains was further investigated by analytical isoelectric focusing (pI). Enzymes with pIs of 5.4, 5.6, 7.4, 7.6 and >8.5 were detected. Sixty-nine percent of the isolates produced a pI 5.4 enzyme that cofocused with TEM-1. Beta-lactamase assays revealed that hyperproduction of these enzymes was the predominant mechanism for clavulanate resistance. Twelve (75%) of the isolates were able to transfer their ampicillin resistance. The ampicillin and ampicillin plus clavulanic acid MIC values of all transconjugants were above 256 microg/ml. Transferable ampicillin resistance was associated with resistance to other antibacterials at the following frequencies: tetracycline 92%, trimethoprim 83%, streptomycin 50%, gentamicin 25%, and chloramphenicol 8%. In conclusion, it has been suggested that commensal bacteria in normal populations make up the largest reservoir of antibiotic-resistant genes. Although the exact molecular mechanisms could not be determined, the current study shows that the incidence of ampicillin and clavulanic acid resistance is also high in commensal fecal flora.

Ampicillin↗
PubMed1997

Relationship between fecal sampling times and sensitivity and specificity of immunochemical fecal occult blood tests for colorectal cancer: a comparative study.

PURPOSE: The present study was conducted to assess the accuracy of three testing methods using an immunochemical fecal occult blood test based on the number of samples as the optimum means for screening of colorectal cancer. METHOD: One hundred eighty-four patients with colorectal cancer and 368 healthy controls served as the subjects for this study. Each subject was tested by an immunochemical fecal occult blood test for three consecutive days. For evaluation of the most desirable number of sampling times, we used the results of the first day for the one-day method, results of the first and second days for the two-day method, and results of three consecutive days for the three-day method. Sensitivities and specificities of the three testing methods were evaluated. RESULTS: Sensitivities of an immunochemical fecal occult blood test were calculated as 67.9 percent for the one-day method, 88 percent for the two-day method, and 90.8 percent for the three-day method; specificity was as follows: 97.5 percent for the one-day method, 95.6 percent for the two-day method, and 92.1 percent for the three-day method. A significant difference in sensitivity was shown between the one-day and two-day and the one-day and three-day methods (P < 0.01); also, a significant difference in specificity was shown between one-day and three-day and two-day and three-day methods (P <0.05). CONCLUSIONS: These results indicate that the two-day method is recommended for immunochemical fecal occult blood testing as a means of screening for colorectal cancer.

Antibodies, Monoclonal↗
PubMed2002

Development, evaluation, and application of lateral-flow immunoassay (immunochromatography) for detection of rotavirus in bovine fecal samples.

A lateral-flow immunoassay (LFT) was developed to detect bovine rotavirus in fecal samples. Using samples (n = 74) from diarrheic calves, a comparison of the LFT with a commercial latex agglutination test (LAT) and transmission electron microscopy (EM) was conducted. When EM was used as the reference method, initial studies of 29 samples indicated 70 and 80% sensitivities of the LFT and LAT, respectively, with both being 100% specific. When the LAT was the reference test, the LFT was 75% sensitive and 91% specific. Additional specimens (n = 45) were tested by the LFT and LAT alone, and results were identical for both methods.

Animals↗
PubMed2003

Escherichia coli O157:H7 prevalence in fecal samples of cattle from a southeastern beef cow-calf herd.

The proportion of fecal samples culture-positive for Escherichia coli O157:H7 was determined for samples collected from 296 beef cows on pasture in a single Florida herd in October, November, and December 2001. The overall proportion of samples that cultured positive was 0.03. The proportion of cows that were culture-positive on at least one occasion was 0.091. No effect of pregnancy status or nutritional regimen on the proportion of culture-positive samples for E. coli O157:H7 was detected. We detected a breed effect on the shedding of E. coli O157, with Romosinuano cows having a lower (P < 0.01) proportion of samples culture-positive than Angus or Brahman cows. This difference might have resulted from the presence of confounding variables; however, it also might represent evidence of breed-to-breed genetic variation in E. coli O157 shedding. Further research is warranted to evaluate breed as a possible risk factor for shedding of this important foodborne pathogen. Further substantiated findings could indicate that breed is a cow-calf-level critical control point of E. coli O157:H7.

Animals↗
PubMed1989

Proportion of beta-D-glucuronidase-negative Escherichia coli in human fecal samples.

Convenient assays and reports that almost all clinical isolates of Escherichia coli produce beta-D-glucuronidase (GUR) have led to great interest in the use of the enzyme for the rapid detection of the bacterium in water, food, and environmental samples. In these materials, E. coli serves as an indicator of possible fecal contamination. Therefore, it was crucial to examine the proportion of GUR-negative E. coli in human fecal samples. The bacterium was isolated from 35 samples, and a mean of 34% and a median of 15% were found to be GUR negative in lauryl sulfate tryptose broth with 4-methylumbelliferyl-beta-D-glucuronide. E. coli from three samples were temperature dependent for GUR production: very weakly positive at 37 degrees C but strongly positive at 44.5 degrees C. These results remind us of differences between fecal and clinical E. coli populations, of diversity in GUR regulation and expression in natural populations of E. coli, and of the need for caution in using GUR for the detection of fecal E. coli.

Enterobacteriaceae↗
PubMed1999

Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples.

A modified procedure was used for culture of Mycobacterium paratuberculosis (Mptb) from bovine feces. Bovine fecal samples were decontaminated with NaOH, exposed to a mixture of oxalic acid and malachite green, incubated in a mixture of neomycin and amphotericin B. Decontaminated specimens were inoculated onto modified Löwenstein-Jensen medium. Specimens processed by high-speed centrifugation showed growth earlier than specimens prepared by low-speed centrifugation. However, the overall number of positive cultures at 16 weeks was not different for the 2 methods. When infected dairy herds were sampled 4 times at 6-month intervals and culture-positive cows were culled, the prevalence of infected cattle declined over time. After selective culling, the cattle left in the herds shed low numbers of Mptb, which explains why it took longer for cultures to become positive. No heifers younger than 11 months were culture positive, but heifers 13-14 months of age were more frequently culture positive than were heifers of any other age. The 16-week culture period is needed with this method to detect cattle shedding low numbers of Mptb. High-speed centrifugation of samples does not increase the efficiency of identification of animals shedding Mptb.

Age Factors↗
PubMed1989

[Electron microscopic virus detection in fecal samples from swine with enteric diseases between 1981 and 1987].

Results of electron microscopical studies of 517 fecal samples from diarrhoeic pigs out of the years 1981 and 1987 were reported. Coronavirus was found in 26.3%, rotavirus in 4.3%, parvovirus in 2.1%, coronavirus like particles in 2.9% and picornavirus like particles in 2.3% of the samples. In some cases (2.3%) double infections could be detected occurring as well between corona- and rotavirus, parvovirus or picornavirus like particles as between rotavirus and parvovirus and picornavirus like particles, respectively. The results were reported annually.

Animals↗
PubMed1989

[The detection of Clostridium botulinum in fecal samples of cattle and swine and in the raw material and animal meal of different animal body rendering plants].

Fecal samples of cattle and swine and samples of raw material and pulverized dehydrated meat taken from three rendering plants were investigated with special enrichment methods on the presence of Clostridium botulinum to get a view about the hygienic risk by the incidence of C. botulinum in rendering plants. Eight-six specimens were examined: 25 fecal specimens each of swine and cattle, 11 of raw material and 25 of pulverized dehydrated meat of three rendering plants. Twelve specimens contained C. botulinum: 7 fecal specimens, 6 of swine and one of cattle, 4 raw material specimens and one of pulverized dehydrated meat. C. botulinum was detected by its toxin production in culture medium. Six times C. botulinum type E, twice C. botulinum type B and one time C. botulinum type C was identified. C. botulinum could not be typed in other cases because the toxin quantities were too small. C. botulinum type E was detected in raw material and pulverized dehydrated meat in one of the three examined rendering plants.

Animals↗
PubMed1994

[Gas chromatographic analysis of C1-C6 fatty acids in rat fecal samples].

A gas chromatography procedure was performed to determine C1-C6 fatty acids in rat feces. Acidated fecal homogenate was extracted with ethyl ether-petroleum ether (1 + 1). After adding tetramethylammonium hydroxide to the extract, the quaternary ammonium salts of the fatty acids could react with ethyl bromide in the presence of N,N-dimethyl acetamide producing corresponding esters. The derived products were separated by the column packed with ethylene glycol succinate polyester and were determined with Flame ionization detector (FID). The extractive efficiencies of C1-C6 fatty acids from the fecal samples were 88.5%-100%, and the recoveries ranged from 82.4% to 110.8% with the coefficients of variation of 3.2%-10.9%. The detection limits were at 1ppm. The optimum conditions of esterification are discussed. The method is safe and rapid, and has been applied to the analysis of C1-C6 fatty acids in rat feces.

Animals↗
PubMed2003

[Occurrence of Cryptosporidium sp in fecal samples of children less than 10 years old with clinical indication of Rotavirus].

The presence of oocysts of Cryptosporidium sp was investigated in 485 fecal samples of children with clinical indication of Rotavirus. No significant differences were observed between Cryptosporidium sp. and rotavirus occurrence and fecal consistency. Cryptosporidium sp also should be performed in the laboratory diagnosis of diarrheic episodes in children.

Animals↗
PubMed1995

Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples.

Three 16S rRNA hybridization probes were developed and tested for genus-specific detection of Bifidobacterium species in the human fecal flora. Variable regions V2, V4, and V8 of the 16S rRNA contained sequences unique to this genus and proved applicable as target sites for oligodeoxynucleotide probes. Determination of the genus specificity of the oligonucleotides was performed by whole-cell hybridization with fluorescein isothiocyanate-labelled probes. To this end, cells were fixed on glass slides, hybridized with the probes, and monitored by videomicroscopy. In combination with image analysis, this allowed quantification of the fluorescence per cell and objective evaluation of hybridization experiments. One of the probes developed was used to determine the population of Bifidobacterium spp. in human fecal samples. A comparison was made with results obtained by cultural methods for enumeration. Since both methods gave similar population estimates, it was concluded that all bifidobacteria in feces were culturable. However, since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.

Base Sequence↗
PubMed1984

Rapid screening of fecal samples for Vibrio cholerae by a coagglutination technique.

A coagglutination procedure for detecting Vibrio cholerae was applied directly to 125 watery fecal samples received in the laboratory for bacteriological culture: many of these were from suspected cases of cholera. Of 47 bacteriologically proved cases of cholera, 44 (93.6%) gave positive results by the coagglutination method. There was a good correlation between the coagglutination method, dark-field microscopy, and culture.

Agglutination Tests↗
PubMedDate not supplied

Evaluation of a rapid screening assay for bacterial identification (Dot-ELISA) in fecal samples from children.

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA.

Child, Preschool↗
PubMed1976

Separation and properties of enterovirus and reovirus recovered from a fecal sample of calf with diarrhea.

In April, 1971, a disease with pyrexia and diarrhea as main symptoms broke out collectively among calves. Fecal samples were collected from calves involved and inoculated into bovine kidney (BK) cell cultures. As a result, the diarrheal feces of one calf were suspected to contain two agents simultaneously. One agent (C-121 E strain) was isolated from the primary infected BK cell culture fluid by terminal dilution passages. It had been predominant in replication and shown a cytopathic effect which gave rise to a granular appearance in the early stage of culture. The other agent (C-121 R strain) was isolated from the primary infected BK cell culture fluid by neutralizing the C-121 E strain contained in this fluid with antiserum against this strain. It caused cytoplasmic inclusion bodies to form. On the basis of their physico-chemical properties, the C-121 E strain was identified as bovine enterovirus and the C-121 R strain as reovirus. Serological tests indicated that some of the affected calves had been infected not only with the two strains isolated, but also with bovine viral diarrhea virus, bovine adenovirus type 7, and bovine parvovirus.

Animals↗
PubMed2006

Mosaic tetracycline resistance genes are widespread in human and animal fecal samples.

Mosaic tetracycline resistance genes comprising tet(O), tet(W), and tet(32) sequences were abundant in DNA extracted from pig and human fecal samples, accounting for 78% (50/64) and 46% (37/80) of genes amplified with a tet(O) primer set, respectively, in two samples. The nonmosaic tet(32) gene was isolated from a human saliva bacterium.

Animals↗