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PubMed1980

Accurate analysis of stable isotopes 68Zn, 70Zn, and 58Fe in human feces with neutron activation analysis.

Accurate isotopic analysis of human fecal samples is discussed in relation to mineral bioavailability studies of human subjects. It is shown that for bioavailability investigations of Zn and Fe, the method of neutron activation analysis is suitable for routine application to clinical experiments; and the human fecal samples resulting from diet suitably enriched in isotopes 70Zn and 58Fe can be routinely analyzed with relative precisions of about 1% for the isotopes 68Zn, 70Zn and 58Fe.

Feces↗
PubMed1996

The characterization of VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses from field samples using RT-PCR and RFLP analysis.

Characterization of the VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses (BRV) from field samples was performed using RT-PCR and RFLP analysis. After RT-PCR amplification of the full length VP7 genes and partial length VP4 genes (nucleotides 1 to 1096), four enzymes, EcoRV, NlaIV, BamHI and HpaII were used for digestion analysis. For VP7, four RFLP profiles were observed after analysis of the digests: they were designated as G6, G6s (subtype, showed about 86% nucleotide and 90% amino acid identity to reference G6 strains), G8 and G10. For VP4, three RFLP profiles were observed: designated as P[1], P[5] and P[11]. The G typing analysis of 86 BRV fecal samples from 5 states, representing at least 11 different herds revealed that 60.5% (52/86) were G6, which included G6s (9/52); 19.8% (17/86) were G10; 7% (6/86) were G8; 10.4% (9/86) were G6 and G10 mixtures including two G6s samples; and 2.3% (2/86) were G6 and G6s mixtures. The P typing analysis of the same 86 fecal samples revealed that 64% (55/86) were P[5]; 28% (24/86) were P[11]; 1.2% (1/86) were P[1] and 6 samples (7%) were mixtures of either P[11] or P[5]. When the same samples were analyzed according to G and P type specificity, all possible combinations of G and P types existed in the field. The G6P[5] type was most prevalent and accounted for 46.7% (41/86) of the samples; 12.8% (11/86) were G10P[11]; 7% (6/86) were G10P[5] and an equal number were G6sP[11]. The G6P[11] (n = 2), G8P[1] (n = 1), G8P[5] (n = 1) and G8P[11] (n = 3) combinations were also observed. The following mixed BRV infections were observed in the field samples; G6sP[5 + 11] (n = 1), G8P[5 + 11] (n = 1), G6 + G10P[5] (n = 1) G6 + G10P[5 + 11] (n = 2), G6 + G6sP[11] (n = 1), G6 + G6sP[1 + 11] (n = 1), G6s + G10P[11] (n = 1) and G6s + G10P[5 + 11] (n = 1). Information on the G and P types and G/P combinations in the field samples should be useful for understanding the epidemiology of BRV and designing vaccination strategies to control BRV in the field.

Animals↗
PubMed2004

Oxalate degradation by intestinal lactic acid bacteria in dogs and cats.

This study evaluated the ability of the lactic acid bacteria (LAB) component of canine and feline feces to degrade oxalate in vitro. Oxalate degradation by individual canine-origin LAB was also evaluated. The effects of various prebiotics on in vitro oxalate degradation by selected oxalate-degrading canine LAB was also evaluated. Canine fecal samples reduced oxalate levels by 78 +/- 12.2% (mean +/- S.D.; range: 44-97%, median: 81%). Feline results were similar, with oxalate reduction of 69.7 +/- 16.7% (mean +/- S.D.; range: 40-96%, median: 73%). Thirty-seven lactic acid bacteria were isolated from canine fecal samples. Mean oxalate degradation was 17.7 +/- 16.6% (mean +/- S.D.; range: 0-65%, median: 13%). No oxalate degradation was detected for four (11%) isolates, and 10/37 (27%) degraded less than 10% of oxalate. The effects of lactitol, arabinogalactan, guar gum, gum Arabic, inulin, maltodextrin or a commercial fructooligosaccharide (FOS) product on in vitro oxalate degradation by five canine LAB isolates were highly variable, even within the same bacterial species. Overall, in vitro degradation was significantly greater with guar gum compared to arabinogalactan (P < 0.05), gum Arabic (P < 0.05), and lactitol (P < 0.01). This study suggests that manipulation of the LAB component of the canine and feline gastrointestinal microflora may decrease intestinal oxalate, and correspondingly intestinal oxalate absorption and renal excretion, thus potentially reducing oxalate urolithiasis.

Animals↗
PubMed1978

Isolation of Treponema hyodysenteriae from sources other than swine.

Fecal samples were collected from animals and environments on 3 swine farms and cultured for Treponema hyodysenteriae. Each farm was a farrow-to-finish operation and, at the time of sampling, swine dysentery was enzootic among 8- to 22-week-old pigs. Pathogenic T hyodysenteriae was isolated from pigs on all 3 farms. On farm A, nonpathogenic T hyodysenteriae was isolated from a sample of lagoon water. On farm B, pathogenic T hyodysenteriae was isolated from a waste-holding pit. On farm C, a dog was observed to be eating feces of pigs that had swine dysentery. The dog was diarrheic and a fecal sample yielded a pathogenic isolant of T hyodysenteriae. Further isolation attempts were unsuccessful after the dog was removed from the infected premises. Isolation of pathogenic and nonpathogenic organisms from waste-holding systems emphasizes the need for cultural techniques in detecting pathogenic T hyodysenteriae.

Animals↗
PubMedDate not supplied

[Control of fecal fermentation with paramomycin and metronidazole].

Using and old procedure to investigate fecal fermentation (FF), modified to make it quantitative a relation was recently found between exaggerated FF and meteorism. FF levels probable reflect the intensity of colonic fermentation. Consequently, it seems desirable to control FF to be able, then, to control exaggerated colonic fermentation in patients with meteorism. In the present communication, a report is made on the effect on FF levels, obtained with fecal samples from 22 patients with meteorism, exerted by the addition of: 1) an antibiotic, paramomycin, with a definite inhibitory capacity on coliform bacteria; 2) a chemotherapeutic agent, metronidazolee, with a definitive inhibitory capacity on anerobic bacteria; and 3) a mixture of paramocycin and metronidazole. Basal FF was normal in 12 out of 22 fecal samples, and exaggerated in the remaining 10 samples. In about all the samples - as well in the samples that rendered normal basal FF as in the samples that rendered exaggerated basal FF - the addition of paramomycin, of metronidazole and of paramomycin and metronidazole in combination significantly reduced the FF levels; and the reductions exerted by each of the additions were not significantly different among each other. Analyzing the obtained results, it was concluded: a) that fecal anaerobic bacteria are, very probably, the main agents of FF, while coliform bacteria only favour the action of the former; and b) that paramomycin, metronidazole or paramomycin and metronidazol in combination could be useful to manage patients with meteorism and exaggerated FF.

Adult↗
PubMed2000

Usefulness of simultaneous detection of toxin A and glutamate dehydrogenase for the diagnosis of Clostridium difficile-associated diseases.

The aim of this study was to evaluate an immunoassay (Triage; Biosite Diagnostics, BMD, France) for detecting both a specific antigen of Clostridium difficile (glutamate dehydrogenase [GDH]) and toxin A. Evaluation of the test was carried out in 304 fecal samples from patients suspected of having Clostridium difficile-associated diseases. The results with GDH and toxin A were compared with those of a culture and cytotoxicity assay (toxin B). The prevalence rates for toxin B-positive and culture-positive fecal samples were 11.2% and 25%, respectively. The sensitivity of the Triage immunoassay was 90.8% for GDH and 79.4% for toxin A. A negative result with both toxin A and GDH was very reliably able to eliminate a diagnosis of Clostridium difficile-associated disease (negative predictive value 99.6%). Triage is a very rapid (20 min) and easy-to-perform test. It could be useful for diagnostic purposes and also for detecting nontoxigenic strains in epidemiogical studies.

Bacterial Proteins↗
PubMed1997

Cryptosporidium muris in dairy cattle in Brazil.

A total of 580 fecal samples from 30 cows and their calves from three dairy farms in São Paulo State, Brazil, was collected monthly over a period of 1 year (April 1994 to March 1995). A combination of water-ether centrifugal sedimentation technique followed by sucrose centrifugal flotation was used for neonatal samples and sucrose centrifugal flotation for the remainder of the samples. Cryptosporidium muris was found in calves from all farms, although the parasite was not detected in the dams. C. muris oocysts were excreted by 14 of the 30 calves, on at least one occasion during the year. Of the 307 calf fecal samples examined, 53 (17.3%) were positive for C. muris. The mean persistency of oocyst shedding in feces was 4 +/- 3 months. Sixty-four per cent of the positive calves excreted few oocysts, while 36% discharged many oocysts for a longer period (P < 0.05). Oocysts were first detected at the mean age of 3 +/- 1 months. The age of the positive calves ranged from 6 weeks to 14 months.

Animals↗
PubMed2005

Salmonella in dairy operations in the United States: prevalence and antimicrobial drug susceptibility.

Salmonella serotypes are important foodborne pathogens of humans that can be acquired through consumption of contaminated meat and dairy products. Salmonella infection also can be a significant animal health issue. As part of a national study of U.S. dairy operations conducted between March and September 2002, fecal samples were collected from representative cows in 97 dairy herds in 21 states and were cultured to determine the prevalence of Salmonella shedding. Salmonella was recovered from the feces of at least one cow in 30.9% of the herds. Overall, 7.3% of fecal samples were culture positive for Salmonella. The three most frequently recovered serotypes were Salmonella Meleagridis (24.1%), Salmonella Montevideo (11.9%), and Salmonella Typhimurium (9.9%). The susceptibilities of Salmonella isolates recovered were determined using a panel of 16 antimicrobial drugs. Salmonella isolates recovered from dairy cows had relatively little resistance to these antimicrobial agents; 83.0% of the isolates were susceptible to all antimicrobials tested. This study provides updated information on the prevalence and susceptibility patterns of Salmonella in dairy herds and on cow and herd characteristics. These data contribute to our understanding of the ecology of Salmonella in the dairy farm environment.

Animals↗
PubMed2005

Risk of infection with Cryptosporidium parvum and Cryptosporidium hominis in dairy cattle in the New York City watershed.

OBJECTIVE: To determine the risk posed by Cryptosporidium parvum and Cryptosporidium hominis from dairy cattle in the New York City watershed (NYCW). SAMPLE POPULATION: Samples from cattle at risk for shedding Cryptosporidium organisms on randomly selected dairy farms in the NYCW. PROCEDURE: Feces were collected for 4 years from calves at risk for infection on 37 dairies. Oocysts were detected by use of centrifugation concentration-flotation microscopy. The DNA was directly isolated from fecal samples and used to amplify fragments of the small subunit ribosomal RNA and thrombospondin-related adhesion protein C-2 genes by use of nested polymerase chain reaction assays. Small subunit ribosomal RNA fragments were restriction digested by the enzyme Vspl and thrombospondin-related adhesion protein C-2 fragments were digested by Eco91l to distinguish between C hominis (formerly known as genotype 1) and C parvum (formerly known as genotype 2). RESULTS: Of 437 fecal samples examined, 214 contained oocysts. Amplicons were generated for 200 samples. We can be certain, with 95% confidence, that cattle in the NYCW did not harbor C hominis. CONCLUSIONS AND CLINICAL RELEVANCE: Cryptosporidium infections in cattle are under examination because of the potential contamination of public waters by manure. Although cattle may be the source of zoonotic infection via C parvum, they pose little risk for C hominis (the strain commonly isolated from humans in waterborne outbreaks of disease). Other sources of oocysts should be considered when investigating outbreaks attributable to contaminated urban drinking water because cattle pose only a small risk via shedding of C hominis.

Animals↗
PubMed2004

Ertapenem pharmacokinetics and impact on intestinal microflora, in comparison to those of ceftriaxone, after multiple dosing in male and female volunteers.

The pharmacokinetics of ertapenem and ceftriaxone were investigated in an open, randomized, two-period crossover study after single- and multiple-dose administration in 10 healthy volunteers (five women and five men). Both antibiotics were administered intravenously once daily for 7 days at dosages of 1 g (ertapenem) and 2 g (ceftriaxone). The concentrations of the antibiotics in serum and urine were quantified by the agar well diffusion method bioassay and, in addition, for ertapenem only, by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). For ertapenem the maximum concentration of the drug in plasma (C(max)) was 256 mg/liter, the half-life was 20.7 h, and the area under the plasma concentration-time curve (AUC) was 830 mg. h/liter. The concentrations in fecal samples were (mean value) 37.2 and 32.7 mg/kg on day 4 and day 8, respectively. Ceftriaxone exhibited a mean C(max) of 315 mg/liter, a half-life of 7.6 h, and an AUC of 1,556 mg. h/liter. The mean concentrations in fecal samples were 153 and 258 mg/kg on day 4 and day 8, respectively. No accumulation of ertapenem or ceftriaxone was detected at steady state. A slightly but significantly decreased AUC for ertapenem was detected for the female volunteers. No serious adverse event was observed. Both antibiotics induced a marked decrease in the anaerobic microflora (4-log-unit decreases in lactobacilli, bifidobacteria, clostridia, and bacteroides) and Escherichia coli, whereas the number of enterococci increased (4 log units). A slight overgrowth of yeasts was observed with both regimens. In all cases the microflora returned to normal levels on days 21 to 35.

Adult↗
PubMed1999

Fecal shedding of feline coronavirus in adult cats and kittens in an Abyssinian cattery.

OBJECTIVE: To determine patterns of fecal shedding of feline coronavirus (FCV) by cats, age at which kittens first began to shed FCV in their feces, and whether there was any relationship between fecal shedding of FCV and serum antibody titers in adult cats or kittens. DESIGN: Prospective observational study. ANIMALS: 15 adult cats and 18 kittens from a single cattery. PROCEDURE: Blood and fecal samples were collected from adult cats every other month for 13 months. Serum FCV antibody titers were measured by use of an indirect immunofluorescence assay. A reverse-transcriptase, nested polymerase chain reaction assay was used to detect FCV in feces. Blood and fecal samples were collected from kittens at approximately 2-week intervals from 3 weeks to 15 weeks of age. RESULTS: Adult cats shed FCV intermittently. All adult cats shed virus in their feces at least once during the year, and 4 of 15 shed virus > 75% of the time. Serum antibody titer was not significantly associated with shedding of FCV. For the kittens, median age at the time FCV was first detected in feces was 67 days (range, 33 to 78 days). All except 1 of the kittens was found to be shedding virus in their feces before or at the time of seroconversion. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that serum FCV antibody titers are not a good indicator of shedding of FCV in the feces. Kittens may shed FCV in their feces before they seroconvert, and all kittens in a cattery in which FCV infection is endemic may be infected before 12 weeks of age.

Animals↗
PubMedDate not supplied

Evaluation of a QIAamp DNA stool purification kit for Shiga-toxigenic Escherichia coli detection in bovine fecal swabs by PCR.

A commercial kit intended for Taq polymerase inhibitor removal was tested to detect Shiga-toxigenic Escherichia coli (STEC) by polymersase chain reaction (PCR) directly from cattle fecal samples. Forty-five samples were analysed for the presence of stx genes. Results were compared to those obtained by two other methods: amplification of DNA purified by a non-commercial procedure (heat lysis protocol), and amplification of DNA from samples cultured in solid media, commonly used in our lab. Identical numbers of positive samples (33/45, 73 %) were obtained with the QIAamp DNA stool purification kit and the culturing procedure, suggesting an adequate removal of inhibitors that interfere in PCR amplification from the feces. Besides, the number of positive samples detected using DNA purified by the non-commercial protocol was lower, 25/39 (64%) than that achieved by using the kit. In conclusion, the use of the QIAamp DNA stool purification kit provided a rapid stx gene detection by PCR in bovine fecal samples.

Animals↗
PubMed1987

Determination of tocopherol and tocopherol acetate concentrations in human feces using high-performance liquid chromatography.

A method of sample clarification and high-performance liquid chromatography (HPLC) specifically developed to permit simple and rapid determination of vitamin E (alpha-tocopherol, E) and vitamin E acetate (EA) in feces is reported. Retinol acetate (RA) was used as an internal standard. The vitamins of interest were extracted from an aqueous stool suspension into an organic phase (ethyl acetate-butanol), which was injected directly onto the reversed-phase HPLC system. An isocratic mobile phase of methanol-water (97:3) was employed, with ultraviolet detection at 275 and 285 nm (to permit simultaneous monitoring and absorbance ratio determination). Recoveries of exogenous RA, E, and EA from stool suspensions (relative to water) were 99.0 +/- 7.0, 100.9 +/- 7.0, and 101.2 +/- 13.3%, respectively (n = 10). The organic matrix could be stored at -35 degrees C overnight with no change in E or EA results. Sensitivities for E and EA were 80 and 102 micrograms/g of stool, respectively. Each analysis required nine min. The within-day coefficients of variation were 2.9, 3.6, and 3.0% (n = 7) for RA, E, and EA, respectively. Neither E nor EA were detected in baseline fecal samples from fourteen subjects, but both were present in high but varied concentrations after four weeks supplementation with oral d,l-EA. E but not EA was present in blood samples drawn during periods of oral supplementation with EA. There was poor correlation between fecal levels of E and EA, and the increase in serum levels of E. This method permits rapid, selective, and precise determination of E and EA in human fecal samples.

Antioxidants↗
PubMed2002

Hormones as indicators of stress.

Animal welfare is of increasing importance and absence of chronic stress is one of its prerequisites. During stress, various endocrine responses are involved to improve the fitness of the individual. The front-line hormones to overcome stressful situations are the glucocorticoids and catecholamines. These hormones are determined as a parameter of adrenal activity and thus of disturbance. The concentration of glucocorticoids (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantage that they can be easily collected and this procedure is feedback free. Recently, enzyme immunoassays (EIA) have been developed and successfully tested, to enable the measurement of groups of cortisol metabolites in animal feces. The determination of these metabolites in fecal samples is a practical method to monitor glucocorticoid production.

Animal Welfare↗
PubMed1999

Detection of shiga-like toxin (stx1 and stx2), intimin (eaeA), and enterohemorrhagic Escherichia coli (EHEC) hemolysin (EHEC hlyA) genes in animal feces by multiplex PCR.

A multiplex PCR was developed for the rapid detection of genes encoding Shiga toxins 1 and 2 (stx1 and stx2), intimin (eaeA), and enterohemolysin A (hlyA) in 444 fecal samples derived from healthy and clinically affected cattle, sheep, pigs, and goats. The method involved non-solvent-based extraction of nucleic acid from an aliquot of an overnight culture of feces in EC (modified) broth. The detection limit of the assay for both fecal samples and pure cultures was between 18 and 37 genome equivalents. stx1 and hlyA were the most commonly encountered virulence factors.

Adhesins, Bacterial↗
PubMed2001

Phenotypic and genotypic characterization of Escherichia coli verotoxin-producing isolates from humans and pigs.

The aim of this study was to characterize verotoxin-producing Escherichia coli (VTEC) isolates obtained from humans and pigs in the same geographic areas and during the same period of time in order to determine whether porcine VTEC isolates could be related to human cases of diarrhea and also to detect the presence of virulence factors in these isolates. From 1,352 human and 620 porcine fecal samples, 11 human and 18 porcine verotoxin-positive isolates were obtained by the VT immunoblot or the individual colony testing technique. In addition, 52 porcine VTEC strains isolated from diseased pigs at the Faculté de médecine vétérinaire during the same period or from fecal samples collected previously isolated at slaughterhouses were characterized in this study. Antimicrobial resistance profiles were different between human and porcine isolates. In general, the serotypes observed in the two groups were different. No porcine isolate was of serotype O157:H7; however, one isolate was O91:NM, a serotype that has been associated with hemorrhagic colitis in humans. Also, one serotype (O8:H19) was found in isolates from both species; however, the O8:H19 isolates of the two groups were of different pathotypes. The pathotypes observed in the human and porcine isolates were different, with the exception of VT2vx-positive isolates; the serotypes of these isolates from the two groups were nevertheless different. Pulsed-field gel electrophoresis analysis indicated no relatedness between the human and porcine isolates. In conclusion, these results suggest that the porcine and human isolates of the present study were not genetically related. Most porcine VTEC isolates did not possess known virulence factors required to infect humans. However, certain non-O157:H7 porcine VTECs may potentially infect humans.

Animals↗
PubMedDate not supplied

[Rotavirus A infections and reinfections: genotyping and vaccine implications].

OBJECTIVE: To identify rotavirus A and the most prevalent G and P genotypes in children with acute diarrhea, and to the describe the occurrence of rotavirus infection and reinfection. METHODS: Group A rotavirus specimens were obtained from fecal samples from children with acute diarrhea in Goiânia, state of Goiás, Brazil from July 2000 to October 2002. Rotavirus A positive children and a control group (children of the same age and sex, without diarrhea and with no evidence of rotavirus in the first fecal samples) were followed prospectively during one year. All rotavirus A positive samples were genotyped using RT-PCR/nested-PCR. RESULTS: A total of 77 group A rotavirus strains (37.2%) were identified in the diarrheic samples of 207 children. The following G genotypes were identified: G1 (62.3%), G9 (34.4%) and G4 (3.3%). With regard to P genotyping, 59% were characterized as P[8], 7.7% as P[6], 23.1% as P[6]+P[8], 7.7% as P[4]+P[8] and 2.6% as P[4]+P[8]. The following associations were observed: G1P[8] (77.8%), G9P[8] (11.1%), G4P[8] (5.6%) and G1P[6] (5.6%). No reinfection was observed in the 40 rotavirus A (+) children. However, but two of 40 children who were initially negative for this agent developed rotavirus infection during the same period. CONCLUSIONS: The predominant G and P genotypes observed were similar to those found in new vaccines. No reinfection occurred during one-year of follow-up for any of the genotypes identified.

Acute Disease↗
PubMed1989

Prevalence of patent Baylisascaris procyonis infection in raccoons (Procyon lotor) in Ithaca, New York.

The prevalence of patent Baylisascaris procyonis infection in raccoons was determined by examining fecal samples collected between July 1986 and May 1987 in Ithaca, New York. September, October, and November had the highest prevalence of infection (35-48%). Significant differences (P less than 0.001) were found when months were grouped by season to test the hypothesis that a fecal sample's probability of being positive does not vary from month to month. Fall was the season contributing most to the overall chi-square statistic. Host sex/age class and prevalence of patent infection were investigated. The raccoons were aged as either juveniles or adults. A significantly higher prevalence of patent infection (P less than 0.001) was found in juveniles when compared to adults. No statistically significant difference was found in other comparisons of host sex and age. Contingency analysis tested the independence of sex/age class/season and presence of eggs. The results of the test were significant (P less than 0.001).

Animals↗