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PubMed1997

Excretion and metabolism of trovafloxacin in humans.

The metabolism and excretion of trovafloxacin was investigated in four healthy male volunteers after a single oral administration of 200 mg of [14C]trovafloxacin (118 microCi). Mean values of 23.1 and 63.3% of the administered dose were recovered in the urine and feces, respectively, after 240 hr. The Cmax of total radioactivity and unchanged trovafloxacin in serum was 3.2 micrograms-equiv/ml and 2.9 micrograms/ml, respectively, and peaked in 1.4 hr. The mean AUC0-infinity for radioactivity and trovafloxacin was 58.2 micrograms-eq.hr/ml and 32.2 micrograms.hr/ml, respectively. This implied that unchanged trovafloxacin constituted 55% of the circulating radioactivity. Urine and fecal samples were analyzed by LC/MS/MS for characterization of the metabolites, and the quantity of each metabolite in the matrices was assessed by means of a radioactivity detector. The profile of radioactivity in urine showed three main metabolites that were identified as the trovafloxacin glucuronide (M1), N-acetyltrovafloxacin glucuronide (M2), and N-acetyltrovafloxacin (M3). The major fecal metabolites were M3 and the sulfate conjugate of trovafloxacin (M4). Analysis of circulating metabolites from pooled serum extracts obtained at 1, 5, and 12 hr indicated that M1 was the major circulating metabolite (22% of circulating radioactivity), whereas M2 and M3 were detected in minor amounts. The results of the present study revealed that oxidative metabolism did not play a significant role in the elimination of trovafloxacin, and phase II conjugation was the primary route of trovafloxacin clearance in humans.

Administration, Oral↗
PubMed1989

Occurrence of Cryptosporidium in home daycare centers in west-central Colorado.

Thirty white middle-class 5 yr olds from home daycare centers were examined for Cryptosporidium. Five fecal samples were collected from each child, over a 3-wk period, concentrated by sedimentation, and stained by a modified acid-fast technique. Nine children were positive (30%), 6 of these were asymptomatic (5 of these were under 2 yr old). Two girls and 1 boy, under 2 yr old, had mild diarrhea within 3 wk prior to fecal collection.

Animals↗
PubMed1999

Rotavirus G-type restriction, persistence, and herd type specificity in Swedish cattle herds.

G-typing of rotavirus strains enables the study of molecular epidemiology and gathering of information to promote disease prevention and control. Rotavirus strains in fecal specimens from neonatal calves in Swedish cattle herds were therefore characterized by using G1 to -4-, G6-, G8-, and G10-specific primers in reverse transcription (RT)-PCR. Fecal samples were collected from one dairy herd (herd A) for 4 consecutive years and from 41 beef and dairy herds (herd B) experiencing calf diarrhea outbreaks. Altogether, 1, 700 samples were analyzed by group A rotavirus enzyme-linked immunosorbent assay, and 98 rotavirus-positive specimens were selected for G-typing by RT-PCR. The effect of herd type, time, geographic region, and clinical symptoms on the G-type distribution was evaluated. Altogether (herds A and B), G10 was found in 59 (60. 2%) fecal specimens, G6 was found in 30 (30.6%) specimens, G3 was found in 1 (1.0%) specimen, and G8 was found in 1 (1.0%) specimen. Seven (7.1%) fecal specimens were not typeable. Herd type specificity in the G-type distribution was demonstrated in the herds in herd B. In the 6 beef suckler herds, only G6 was detected, while rotavirus strains from the 35 dairy herds were predominantly (54%) G10. The G-type distribution was restricted in herds A and B. Twenty-nine of 30 strains from herd A were characterized as G10. In the vast majority of herds in herd B, a single G-type was identified. The serotype G10 and the electropherotype persisted over time in herd A. No characteristic G-type variation in the geographic distribution of cattle herds in herd B was obvious. There was no difference in the G-type distributions between the strains from clinically and subclinically rotavirus-infected calves in dairy herd A. The results from this study strongly indicate a pronounced stability in the rotavirus G-type distribution in Swedish cattle herds, which emphasizes the importance of continuous preventive measures for control of neonatal calf diarrhea. A future bovine rotavirus vaccine in Sweden should contain G10 and G6 strains.

Animals↗
PubMed2004

Evaluation of a larval development assay (DrenchRite) for the detection of anthelmintic resistance in cyathostomin nematodes of horses.

A larval development assay (LDA, DrenchRite) was evaluated to determine the effectiveness of this method in detecting anthelmintic resistance in cyathostomin nematodes of horses. A total of 15 horse farms from Georgia and South Carolina (USA) and Population S ponies from the University of Kentucky (USA) were included in this study. Nematode eggs were extracted from pooled fecal samples and placed into the wells of a DrenchRite plate for testing against thiabendazole (TBZ), levamisole (LEV) and 2 ivermectin (IVM) analogs (IVM-1, IVM-2). After a 7-day incubation larvae in each well were counted and data were analyzed by logistic regression. Resistance status of each farm for different drugs was determined in a separate study using a fecal egg count reduction test. LDA were performed on the 15 farms once, however, the Population S cyathostomins were assayed on 3 separate occasions to estimate the consistency of results between assays. Mean TBZ LC50 for oxibendazole resistant, suspected resistant and sensitive farms were 0.2015, 0.1625, and 0.1355 microM, respectively. For LEV, mean LC50 for PYR resistant, suspected resistant and sensitive farms were 1.590, 1.8018 and 1.4219 microM, respectively. All 15 farms had worms susceptible to IVM; mean LC50 for IVM-1 and for IVM-2 were 7.5727 and 87.9718 nM, respectively. A linear mixed model was fitted to the data to determine the relationship between LC50 and LC95 and resistance status for each farm. No meaningful relations were found. Consistency of assays varied between drugs, being best for TBZ and worst for LEV and IVM-1. All farms in this study had benzimidazole-resistant nematodes; therefore usefulness of DrenchRite for discriminating susceptibility versus resistance to this drug class could not be accurately assessed. Moreover, since all farms tested were sensitive to IVM and resistance to this drug class has not yet been reported in cyathostomins, it is not possible to assess accurately the usefulness of DrenchRite LDA for detecting IVM resistance at this time. Assay results for LEV suggest that LEV in a LDA does not yield data that is useful in estimating PYR efficacy in vivo. Based on results for PYR/LEV, the current high prevalence of benzimidazole resistance, no known cases of IVM resistance, and the sometimes extreme variation in results seen in many of the assays, DrenchRite LDA cannot be considered a useful tool for the diagnosis of resistance in cyathostomins of horses at present.

Animals↗
PubMed2004

Evaluation of universal pre-enrichment broth for isolation of Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes from dairy farm environmental samples.

Use of universal pre-enrichment broth (UPB) as a primary enrichment medium for detection of Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes from dairy farm environmental samples was evaluated. There were no differences in bacterial growth between UPB and selective primary enrichment broths for each pathogen inoculated individually or in combination at 10(1) and 10(2) colony forming units/mL. In addition, no differences were observed when UPB and selective primary enrichment broths were compared for detection efficiency of pathogens in artificially contaminated raw milk and fecal samples. Listeria enrichment broth (LEB) was compared with UPB to support growth of L. monocytogenes from naturally contaminated environmental samples. Listeria monocytogenes was isolated from seven of 30 samples enriched in UPB and six of 30 samples enriched in LEB. Dairy farm environmental samples were examined for recovery of the three pathogens using UPB. Subsequent isolation was achieved using selective secondary enrichment of each pathogen. Listeria monocytogenes, Salmonella spp., and E. coli O157:H7 were isolated in 13.4% (30 of 224), 8.9% (20 of 224), and 2.2% (five of 224) of samples, respectively. Isolation rates of the three pathogens were somewhat higher than in previous reports. Overall, UPB supported growth of test pathogens to detectable levels within 24 h. Our results demonstrate that UPB has potential for routine use in isolation of foodborne pathogens from diverse environmental samples.

Animals↗
PubMedDate not supplied

Identification of Cryptosporidium spp. oocysts in fecal smears stained with Heidenhain's iron hematoxylin.

There is no paucity of methods for diagnosing Cryptosporidium spp. infection. The merits of immunoassays notwithstanding, microscopic identification of Cryptosporidium spp. oocysts in fecal samples remains an important diagnostic procedure. It owes the persistence of its use to such characteristics as dispensing with expensive equipment and kits, requiring only basic laboratory facilities, and having a low probability of false positive results when permanent slides are prepared, which can be re-examined in case of doubt. Cryptosporidium spp. oocysts can be readily identified in fecal smears prepared according to a regressive iron hematoxylin staining technique. The number of steps and their duration, as well as costs, were reduced to a minimum without loss of image quality and permanence of the preparations.

Animals↗
PubMed2005

Safety assessment of potential lactic acid bacteria Bifidobacterium longum SPM1205 isolated from healthy Koreans.

The safety assessment of Bifidobacterium longum SPM1205 isolated from healthy Koreans and this strain's inhibitory effects on fecal harmful enzymes of intestinal microflora were investigated. The overall safety of this strain was investigated during a feeding trial. Groups of SD rats were orally administered a test strain or commercial reference strain B. longum 1 x 10(9) CFU/kg body weight/day for four weeks. Throughout this time, their feed intake, water intake and live body weight were monitored. Fecal samples were periodically collected to test harmful enzyme activities of intestinal microflora. At the end of the four-week observation period, samples of blood, liver, spleen, kidney, and gut tissues were collected to determine for hematological parameters and histological differences. The results obtained in this experiment demonstrated that four weeks of consumption of this Bifidobacterium strain had no adverse effects on rat's general health status, blood biochemical parameters or histology. Therefore, it is likely to be safe for human use. Fecal harmful enzymes such as beta-glucosidase, beta-glucuronidase, tryptophanase and urease, were effectively inhibited during the administration of the B. longum SPM1205. These results suggested that this B. longum SPM 1205 could be used for humans as a probiotic strain.

Adult↗
PubMed1980

Ecological investigation on intestinal bacterial flora of patients nursed in protected environmental units.

The present results of investigations performed on fresh fecal samples from a healthy adult and subjects with acute leukemia cultured bacteriologically using both aerobic and anaerobic techniques have led to the following conclusions. 1) In studies on the intestinal bacterial flora of the subjects examined, obligate anaerobic bacteria rather than facultative anaerobes accounted for more than 99% of the microorganisms. 2) Although some qualitative differences seem to exist between fecal bacterial flora components of the normal individual and leukemia patients examined, further investigations considering various factors controlling and influencing the composition of intestinal bacterial flora are needed to evaluate the results obtained. 3) The results obtained herein indicate that changes in intestinal bacterial flora induced by non-absorbable antibiotics are associated with the host defense mechanisms. 4) The relative predominance of aerobic versus anaerobic bacterial flora in the intestine may reflect the degree of severity of clinical manifestation in leukemia patients. In the data obtained from the present investigations, the predominating fecal anaerobic bacteria may serve as a base line for the evaluation and interpretation of the host defense system by patterns of the components of bowel flora brought about by internal or external environmental changes.

Acute Disease↗
PubMed1990

Enzyme-linked immunoassay for detection of Cryptosporidium antigens in fecal specimens.

Cryptosporidium sp. is a ubiquitous 4- to 6-micron protozoan parasite infecting the intestinal tract of humans. It causes mild to fulminant diarrhea in patients, especially immunocompromised persons, and it may be hard to detect by microscopic fecal examination. An indirect, double-antibody enzyme-linked immunosorbent assay (ELISA) was developed using specifically produced goat and rabbit antisera to detect Cryptosporidium antigens in human feces. Of 62 frozen stools from patients with cryptosporidiosis, as detected by at least two microscopic diagnostic techniques, 51 were positive by ELISA; all ELISA-negative specimens came from patients with fewer than five oocysts per 0.01 ml of concentrated fecal sample examined after modified acid-fast or fluorescent monoclonal antibody staining. A total of 182 specimens from persons without Cryptosporidium infection were negative by ELISA in 176 instances; 3 ELISA-positive specimens came from patients with cryptosporidiosis diagnosed earlier. The sensitivity of the assay was 82.3%, and specificity was 96.7%. The predictive value of a positive ELISA was 89.5%, and the predictive value of a negative ELISA was 94.2%. The ELISA was not affected by the presence of eight other intestinal parasites but was sometimes affected by repeated freezing and thawing of fecal specimens. All fecal specimens were heated to 100 degrees C for 2 min to reduce proteolytic enzyme activity, although the necessity of this step needs further evaluation. This first-generation ELISA is a simple, rapid, easily standardized test for Cryptosporidium antigens in stool samples which will be useful for diagnosis and for large-scale epidemiologic studies.

Animals↗
PubMed2005

Use of the duplex TaqMan PCR system for detection of Shiga-like toxin-producing Escherichia coli O157.

Real-time PCR assays have been applied for the detection and quantification of pathogens in recent years. In this study two combinations of primers and fluorescent probes were designed according to the sequences of the rfb(Escherichia coli O157) and stx2 genes. Analysis of 217 bacterial strains demonstrated that the duplex real-time PCR assay successfully distinguished the Escherichia coli O157 serotype from non-E. coli O157 serotypes and that it provided an accurate means of profiling the genes encoding O antigen and Shiga-like toxin 2. On the other hand, bacterial strains that lacked these genes were not detected by this assay. The quantitative ranges of the real-time PCR assay for these two genes were linear for DNA concentrations ranging from 10(3) to 10(9) CFU/ml of E. coli O157:H7 in pure culture and milk samples. The real-time PCR allowed the construction of standard curves that facilitated the quantification of E. coli O157:H7 in feces and apple juice samples. The detection sensitivity of the real-time PCR assay ranged from 10(4) to 10(9) CFU/g (or 10(4) to 10(9) CFU/ml) for feces and apple juice and 10(5) to 10(9) CFU/g for the beef sample without enrichment. After enrichment of the food samples in a modified tryptic soy broth, the detection range was from 10(0) to 10(3) CFU/ml. The real-time PCR assays for rfb(E. coli) (O157) and stx2 proved to be rapid tests for the detection of E. coli O157 in food matrices and could also be used for the quantification of E. coli O157 in foods or fecal samples.

Adult↗
PubMed2003

Enrichment for isolating Salmonella Choleraesuis and other Salmonella spp. from pigs.

The growth of Salmonella Choleraesuis was examined in Rappaport Vassiliadis broth (RV) and Hajna-tetrathionate broth (HTT) at 37 and 42 degrees C. As the enrichment in RV at 37 degrees C was satisfactory for isolating S. Choleraesuis, we used this enrichment for isolation from the samples collected from 15 asymptomatic pigs reared on a S. Choleraesuis contaminated farm. S. Choleraesuis was frequently isolated from six pigs (40.0%) under field conditions. The isolation of other Salmonella serovars than S. Choleraesuis was attempted by using both RV enrichment at 37 degrees C and HTT enrichment at 42 degrees C. Salmonella organisms were isolated from 156 (44.8%) of 348 fecal samples and more frequently with HTT at 42 degrees C (43.4%) than with RV at 37 degrees C (20.9%). If other serovars in addition to S. Choleraesuis are to be surveyed, HTT enrichment should be used in combination with RV enrichment.

Animals↗
PubMed2005

Effect of water sprinkling on incidence of zoonotic pathogens in feedlot cattle.

Heat stress and dusty conditions are common challenges for cattle during the summer, and a typical method of alleviating these problems involves sprinkling cattle and pens with water. The effect of sprinkling water on the incidence of zoonotic pathogens has not been previously studied. Four pens of heifers (n = 41) were cooled using sprinklers, and four pens (n = 43) served as controls. Heifers were crossbred Charolais, with white and red hair coats. Sprinkling was initiated when cattle were on full concentrate feed (July). Fecal samples, hide swipes, and BW were collected on d 0, 28, 63, 95, and 98. Average daily gain, DMI, and G:F were calculated, and carcass traits were collected 36 h after processing. Performance data were analyzed as a randomized complete block design, and zoonotic pathogen data were analyzed using chi2 analysis. Sprinkling tended (P = 0.054) to increase the incidence of fecal Salmonella spp. populations on d 98, but simultaneously tended to decrease (P = 0.058) the Escherichia coli O157:H7 incidence on hides on d 98. The most prevalent Salmonella serovars in this study were Kentucky, Muenster, Meleagridis, and Cerro. Performance measures and carcass traits did not differ between treatments (P > 0.10). Under our conditions, sprinkling cattle with water did not affect the incidence of zoonotic pathogens in feces or on hides.

Animal Husbandry↗
PubMed1979

Course of antibiotic sensitivities in Escherichia coli and Staphylococcus aureus from animals.

In order to establish the evolution of resistance against the common antibiotics from the beginning of their use up to the present time, the sensitivity of Escherichia coli isolated from the cecum of wild boars was determined. The MIC's of E. coli from these animals in a zoological garden was examined for comparison. The samples from the wild boars were collected in five forests in the Belgian Ardennes. The MIC's obtained (Table 1) are comparable to the mean values given in the early literature concerning strains of human origin (Table 7). E. coli from wild boars in a zoological garden has less sensitivity than from domesticated animals. However, the MIC's of staphylococci from fecal matter of the animals in captivity were not markedly enhanced except for streptomycin (Table 3). It was striking to find that staphylococci were absent in all the fecal samples from the wild boars living in their natural environment. The specific habitat of staphylococci is in man. Wild boars are the ancestors of domestic pigs. The MIC's of E. coli from the latter were determined for evaluation of altered resistance. All isolated strains were fairly resistant to the six substances tested (Table 2). Chloramphenicol was still the most active antibiotic. Staphylococci from some domestic animals (pigs, cattle, chicken) were sensitive to all the substances tested except the aminoglucosides and ampicillin (Table 3). E. coli from pigs bred on large farms in Spain (Toledo and Badajoz Provinces) were also highly resistant (Table 4) to five compounds and sensitive to cephalotin, polymyxin B, furoxone, and Carbadox. The samples obtained from family farms also contained unsensitive strains (Avila Province). These animals had never been given antibacterial feed additives as was confirmed by examination of feed samples (Table 6). Staphylococci when present in the Spanish samples can be considered to be antibiotic sensitive with the same exceptions as in the Belgian samples and also unsensitive to chloramphenicol. The use of antibiotics in animal feed was very common in Spain as was shown by feed analysis (Table 6). The risk of transferable resistance by plasmids between animal species must not be very high. In some cases where resistance formation could be mediated by R-factors, sensitive E. coli or staphylococci were isolated. Resistance was not generalized among animals from the same farm or in adjacent sties. No explanation for this discrepancy was found.

Animal Feed↗
PubMed2006

Potential of antimicrobial agents to inhibit fungi to aid isolation of Campylobacter.

In preliminary studies conducted to isolate thermophilic Campylobacter spp. from cull dairy cow fecal samples, growth of fungal strains on Abeyta-Hunt-Bark agar and charcoal cefoperazone desoxycholate agar (CCDA) interfered with isolation of target bacteria. Concentrations of antifungal substances in the agar media were not effective in suppressing growth of antibiotic resistant fungi. The objective of the current study was to determine the efficacy of supplemental antimicrobial agents to inhibit growth of fungi of fecal origin on Campylobacter isolation media where microbial growth is overwhelmed by fungi. The inhibitory effect of rifampicin and seven antimycotic agents--cycloheximide, 5-fluorouracil, 5-fluorocytosine, sodium propionate, potassium sorbate, nystatin, and amphotericin B--was evaluated. Antimicrobial agents were added singly or combined at concentrations ranging from 12.3 to 227 mg/L. Incubation was conducted at 42 degrees C for 24-48 h in gas jars (CO2+H2). The most effective antibiotic combinations that inhibited fungal growth without affecting Campylobacter growth were CCDA selective supplement SR155E with amphotericin B (59.5 or 114 mg/L) or a mixture of rifampicin, 5-fluorocytosine, and nystatin (119 or 227 mg/L each). Results of the present study indicate that addition of supplemental antifungal agents in the presence of antibiotics such as rifampicin may enhance suppression of fungal overgrowth on Campylobacter isolation agar media.

Animals↗
PubMed2005

Gastrointestinal parasites of the colobus monkeys of Uganda.

From August 1997 to July 2003, we collected 2,103 fecal samples from free-ranging individuals of the 3 colobus monkey species of Uganda-the endangered red colobus (Piliocolobus tephrosceles), the eastern black-and-white colobus (Colobus guereza), and the Angolan black-and-white colobus (C. angolensis)--to identify and determine the prevalence of gastrointestinal parasites. Helminth eggs, larvae, and protozoan cysts were isolated by sodium nitrate flotation and fecal sedimentation. Coprocultures facilitated identification of helminths. Seven nematodes (Strongyloides fulleborni, S. stercoralis, Oesophagostomum sp., an unidentified strongyle, Trichuris sp., Ascaris sp., and Colobenterobius sp.), 1 cestode (Bertiella sp.), 1 trematode (Dicrocoeliidae), and 3 protozoans (Entamoeba coli, E. histolytica, and Giardia lamblia) were detected. Seasonal patterns of infection were not apparent for any parasite species infecting colobus monkeys. Prevalence of S. fulleborni was higher in adult male compared to adult female red colobus, but prevalence did not differ for any other shared parasite species between age and sex classes.

Animals↗
PubMed2004

Fecal immunoglobulin A antibodies in dogs infected or vaccinated with canine coronavirus.

Fecal secretory immunoglobulin A (IgA) antibodies in dogs infected or vaccinated with canine coronavirus (CCV) were evaluated by an enzyme-linked immunosorbent assay. The study was carried out with 32 fecal samples collected just before inoculation and at 28 days postinoculation. Five groups were studied: naturally infected dogs, experimentally infected dogs, dogs inoculated with a modified live (ML) CCV vaccine by the intramuscular route, dogs inoculated with an ML CCV vaccine by the oronasal route, and dogs given an inactivated CCV vaccine. Both the naturally and the experimentally infected dogs developed high levels of fecal IgAs. Interestingly, dogs inoculated with the ML CCV vaccine by the oronasal route developed levels of fecal IgA that were higher than those observed in the dogs inoculated with the same CCV vaccine by the intramuscular route or those observed in dogs inoculated with the inactivated vaccine. A relationship between the level of fecal IgAs to CCV and the degree of protection against CCV infection was observed.

Animals↗
PubMed1991

Immunomagnetic separation and DNA hybridization for detection of enterotoxigenic Escherichia coli in a piglet model.

Enterotoxigenic Escherichia coli (ETEC) strains were detected by stool blot hybridization assays using different oligonucleotide probes for the colonization fimbrial antigen F4, heat-stable enterotoxin I (ST I), and heat-labile enterotoxin (LT I) genes. Forty-eight fecal samples and seven samples of intestinal content from ETEC-challenged newborn piglets were processed in two ways: (i) by direct inoculation of bacterial suspension onto nylon membranes overlaying blood agar and (ii) by immunomagnetic enrichment of F4+ ETEC using magnetic beads coated with F4 monoclonal antibodies before inoculation onto nylon membranes. In samples obtained from nondiarrheic piglets pre- and postchallenge, E. coli genes for F4, ST I, and LT I could be detected only after immunomagnetic enrichment. No difference between the two methods in detection of these E. coli genes was observed when stool blots from diarrheic piglets were examined. By using magnetic separation, it was easy to decrease background bacterial flora, intestinal cells, and fecal debris and thus produce purer specimens. The method evaluated in this animal model appeared simple and quick and increased the sensitivity of detection of ETEC strains 100-fold compared with the direct stool blot hybridization assays. Prior bacterial isolation and identification were not necessary.

Animals↗
PubMed1992

Daily variability of equine fecal strongyle egg counts.

Fecal egg counts often are used for diagnosing equine strongyle infections and estimating the number of eggs shed in the feces. An individual egg count should be interpreted in view of the normal fluctuation of egg numbers in an individual horse. In this study, the daily variability of strongyle fecal egg counts from horses was investigated. The Cornell-McMaster egg-counting technique was used to estimate the eggs per gram of feces in repeated daily fecal samples from 39 horses. The variation of the daily egg counts across 4 days was greater than would be expected if a consistent number of eggs were produced and dispersed randomly in the feces. The means and variances of the daily counts from each horse had a logarithmic relationship. For practical purposes, however, the fluctuation of egg counts is low enough for the fecal egg count to be used to identify horses for treatment, to estimate pasture contamination, or to assess response to therapy.

Animals↗