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PubMedDate not supplied

Milk casein-based diet containing TGF-beta controls the inflammatory reaction in the HLA-B27 transgenic rat model.

BACKGROUND: A casein-based formula containing TGF-beta has been successfully used in adolescents during acute episodes of Crohn's disease. The role played by this molecule requires confirmation. We have examined the capacity of a TGF-beta containing diet to control the intestinal inflammation in HLA-B27 transgenic rats, and compared its effects with a similar diet devoid of TGF-beta. METHODS: Three groups of rats were studied. HLA-B27/hbeta2M transgenic rats were fed with a casein-based rat-adapted diet containing TGF-beta or a control casein-based diet without TGF-beta. Fischer control animals were fed the latter. Body weight, dietary intake, tissue weights, fecal samples, leukocyte counts, and acute phase response were analyzed. Intestinal inflammation was assessed by histology, myeloperoxidase, and mRNA expression of cytokines. MUC2 protein expression was assessed by immunohistochemistry. Breakdown of muscle protein was examined. RESULTS: The test diet improved diarrhea increasing the fecal dry matter and the colonic inflammation as shown by a lower inflammatory score (2.43 +/- 1.13 vs 4.42 +/- 0.53, p < .05), lower mucosal thickness (431.25 +/- 72.29 vs 508.57 +/- 81.32 microm, p = .08) and decreased IFNgamma mRNA expression. MUC2 protein expression was increased in HLA rats fed the TGF-beta diet compared with HLA rats fed the control diet, but restitution to normal pattern was not observed. The test diet also decreased leukocytosis and the acute phase response and improved the muscle catabolic response. CONCLUSION: The TGF-beta containing diet has a beneficial effect in an animal model of intestinal inflammation. Our observations support a potential role for dietary TGF-beta in the restoration of immune homeostasis.

Acute-Phase Reaction↗
PubMed1994

A survey of G6 and G10 serotypes of group A bovine rotaviruses from diarrheic beef and dairy calves using monoclonal antibodies in ELISA.

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G10-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Short-chain fatty acid and organic acid concentrations in feces of 10 human volunteers and their correlation with anaerobe cultural over a 15-month period.

Short-chain fatty acid (SCFA) and organic acid concentrations were determined with gas-liquid chromatography (GLC) in fresh feces from 10 healthy volunteers, simultaneously with their fecal cultural counts. To find a simple and reliable method to detect disturbances in fecal flora, we calculated the intraindividual correlations between the concentrations determined by GLC and the fecal cultural counts. Twenty-three (4.7%) significant (P less than 0.05) linear correlations were thus found in nine of the volunteers. Using multivariate analysis, we attempted to assign fecal samples to individuals in accordance with their SCFA, cultural count, and organic acid profiles. Percentages of samples thus classified correctly were 49, 39, and 29, respectively. We conclude from this study that these percentages represent the normal situation in a group of healthy humans and that the low percentage of significant correlation coefficients shows that the relation between fecal SCFA/organic acid concentrations and fecal cultural counts has not been established.

Acids↗
PubMed2003

Detection of transgenic and endogenous plant DNA in rumen fluid, duodenal digesta, milk, blood, and feces of lactating dairy cows.

The objective was to determine the presence or absence of transgenic and endogenous plant DNA in ruminal fluid, duodenal digesta, milk, blood, and feces, and if found, to determine fragment size. Six multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas received a total mixed ration. There were two treatments (T). In T1, the concentrate contained genetically modified (GM) soybean meal (cp4epsps gene) and GM corn grain (cry1a[b] gene), whereas T2 contained the near isogenic non-GM counterparts. Polymerase chain reaction analysis was used to determine the presence or absence of DNA sequences. Primers were selected to amplify small fragments from single-copy genes (soy lectin and corn high-mobility protein and cp4epsps and cry1a[b] genes from the GM crops) and multicopy genes (bovine mitochondrial cytochrome b and rubisco). Single-copy genes were only detected in the solid phase of rumen and duodenal digesta. In contrast, fragments of the rubisco gene were detected in the majority of samples analyzed in both the liquid and solid phases of ruminal and duodenal digesta, milk, and feces, but rarely in blood. The size of the rubisco gene fragments detected decreased from 1176 bp in ruminal and duodenal digesta to 351 bp in fecal samples.

Animals↗
PubMed1980

Rotavirus infection in foals.

Fecal samples from 86 foals with diarrhea were examined by electron microscopy during a 2.5 year period. Of these, 26 (30%) were positive for rotavirus. All of the cases were found in epizootic areas. The disease was produced in an experimental foal by inoculation via stomach tube of a bacteria-free fecal filtrate containing rotavirus. Examination of postmortem tissues from the duodenum and jejunum of 2 naturally infected foals and an experimentally infected foal revealed replicating virus in the intestinal epithelial cells. A limited survey of complement-fixing antibody to rotavirus in horses from Kentucky, Virginia, and France indicated that all horses had antibody to the virus. The sole exception was 1 foal from which blood samples were collected prior to suckling. These results were presumptive evidence that rotavirus is a major cause of diarrhea in foals, and the presence of antibody in horses from diverse areas is evidence for the ubiquitousness of this infection.

Animals↗
PubMed1978

Evaluation of a method for transportation and preservation of anaerobic microorganisms.

Fecal samples from sixty patients were utilized to evaluate a method for the preservation of anaerobic organisms. Comparison was made with a method commonly employed in the collection of specimens from the operating room and bedside. Analysis of the numbers ae preservation of anaerobic fecal flora in a reliable manner during a 24 hour delay between specimen collection and plating.

Bacteriological Techniques↗
PubMed1999

Evaluation of TaqMan PCR assay for detecting Salmonella in raw meat and shrimp.

We evaluated the TaqMan Salmonella amplification/detection kit from PE Applied Biosystems, which uses a polymerase chain reaction (PCR) assay for rapid detection of Salmonella in food samples. This system uses the 5' nuclease activity of Taq DNA polymerase, which digests an internal fluorogenic probe to monitor the amplification of the target gene. The system's sensitivity and specificity were evaluated using 42 serotypes of 68 Salmonella strains isolated from fecal samples from patients in Tokyo, Japan, and 39 non-Salmonella strains in 22 genera. There were no false-negative or false-positive results. This PCR assay can detect 3 CFU per PCR tube of Salmonella in pure culture (120 CFU/ml of TSB culture). PCR signals were attenuated with artificially contaminated shrimp, but a similar detection limit was obtained. TaqMan's performance was tested with 100 meat and chicken samples purchased from stores in Tokyo. Overall, two of the DNA extraction protocols (the Chelex and EnviroAmp methods) worked equally well, with some exceptions. Of the 100 samples analyzed, 10 were positive for Salmonella with both conventional culture methods and the kit and 89 were negative with both. One sample was negative by the culture method but positive by the kit assay. These results indicate that TaqMan is a reliable and rapid method for Salmonella analysis in the food industry. With this system, food samples can be analyzed for Salmonella in less than 20 h.

Animals↗
PubMed1997

Nucleic acid amplification for rapid detection of Rhodococcus equi in equine blood and tracheal wash fluids.

OBJECTIVE: To evaluate the ability of nucleic acid amplification techniques to detect Rhodococcus equi in equine buffy coat, blood, and tracheal wash fluid and to differentiate between virulent and avirulent strains of the bacteria. SAMPLE POPULATION: Blood anticoagulated with EDTA and tracheal wash fluid from healthy horses. PROCEDURE: Logarithmic dilutions of virulent and avirulent strains of R equi were added to equine buffy coat and tracheal wash fluid samples. The DNA was extracted and amplified by polymerase chain reaction (PCR), using primers specific for the 16S ribosomal subunit gene and the virulence plasmid of R equi. RESULTS: PCR with 16S ribosomal subunit primers amplified a 441-bp segment of DNA from virulent and avirulent strains of R equi, but not from samples containing other species of bacteria. The virulence plasmid primers amplified an 875-bp segment of DNA from virulent strains of R equi, but not from avirulent R equi, or from other species of bacteria. Virulent strains of R equi could be identified by PCR and differentiated from avirulent strains within 12 to 24 hours after sample collection, with as few as 10 to 100 organisms present. CONCLUSIONS: PCR can be used to rapidly and accurately identify R equi in equine blood and tracheal wash fluid samples and can differentiate between virulent and avirulent strains of the organism. CLINICAL RELEVANCE: Because PCR can confirm a diagnosis of R equi infection in horses more rapidly and specifically than use of standard culture techniques, extrapolation of this assay to soil and fecal samples could be useful in epidemiologic studies and studies of environmental disinfection or decontamination.

Actinomycetales Infections↗
PubMed1991

Dilatation and closing anal reflexes. Description and clinical significance of new reflexes: preliminary report.

The present communication describes new reflexes which are called 'dilatation and closing anal reflexes', and discusses their clinical significance. The study comprised 21 healthy volunteers and 15 incontinent patients (7 with partial fecal incontinence and 8 with urinary stress incontinence). The technique comprised the introduction into the rectal neck of a balloon-tipped catheter. The balloon was inflated with air in increments of 10 ml up to 50 ml, and the EMG response of the external and urethral sphincters to balloon inflation and deflation was recorded. A new device called 'switch inflation' apparatus was used to inflate the balloon simultaneously with switching of the EMG apparatus. Rapid rectal neck inflation and deflation evoked external anal and urethral sphincter contraction. Slow and gradual inflation or deflation did not initiate the response. The anesthetized external anal sphincter did not respond to the stimulus, while the saline-infiltrated sphincter responded. The latency of the reflexes was recorded. In fecal incontinent patients, the external anal sphincter, on rapid rectal neck inflation or deflation, showed lower EMG activity and longer latency than in normal volunteers; the external urethral sphincter responded as in normal volunteers. In urinary stress incontinent patients, the external anal sphincter responded normally for both rectal neck inflation and deflation. The external urethral sphincter showed lower EMG activity and prolonged latency than normal on rectal neck inflation; it did not respond to deflation. The dilatation and closing reflexes seem to play a role in fecal and urinary continence as well as in fecal sampling. Detectable changes in latency or amplitude of the evoked response indicate a defect in the reflex pathway.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1996

Trypanosoma cruzi: study of the distribution of two widespread clonal genotypes in Bolivian Triatoma infestans vectors shows a high frequency of mixed infections.

The detection of two widespread Trypanosoma cruzi clonal genotypes (20 and 39) in feces of Bolivian specimens of the vector Triatoma infestans was performed by a combination of polymerase chain reaction and clone-specific DNA hybridization. The hybridization pattern of 186 PCR positive samples cf T. infestans feces collected in two Bolivian departments identified clone 20 in 74.2% and clone 39 in 63.4% of the triatomine bugs. For the first time, a high percentage (mean: 43.2 +/- 26%) of mixed infections (presence of both clones in a given fecal sample) in various localities was recorded. Results were in agreement with the two assumptions of independent transmission of clones 20 and 39 and of the absence of selection in the natural cycles under survey. Statistical analysis of the geographical distribution of clones 20 and 39 favored the hypotheses that the frequencies of T. cruzi natural clones are different among localities and that these differences are not proportional to the distances that separate the localities. The epidemiological significance of these results is discussed.

Animals↗
PubMed1995

Protracted viremia during acute sporadic hepatitis E virus infection.

BACKGROUND/AIMS: Hepatitis E virus (HEV) is associated with epidemic and sporadic hepatitis in developing countries. The disease is largely self-limited with no long-term sequelae. The source of HEV for maintenance of the disease in an endemic area is unknown. This study investigated the occurrence and duration of viremia in patients with acute sporadic HEV infection. METHODS: In 26 of 37 patients with sporadic acute non-A, non-B viral hepatitis, HEV infection was diagnosed based on positivity for immunoglobulin M anti-HEV and/or presence of viremia as shown by reverse-transcription polymerase chain reaction. In 4 patients, fecal samples were analyzed for presence of virus using polymerase chain reaction. Multiple samples were studied at varying times in 20 patients. RESULTS: Viremia was detected in 19 of 26 patients. Two patients had viremia in the absence of immunoglobulin M anti-HEV. Four patients had protracted viremia of 45-112 days' duration. One patient showed fecal virus shedding up to the 52nd day of illness. CONCLUSIONS: Protracted viremia and prolonged fecal shedding of HEV were shown in a small group of patients. These patients may serve as temporary virus carriers responsible for continuous contamination of the sewage water.

Acute Disease↗
PubMed2005

Effects of environmental conditions, human activity, reproduction, antler cycle and grouping on fecal glucocorticoids of free-ranging Pampas deer stags (Ozotoceros bezoarticus bezoarticus).

In this study, a commercial enzyme immunoassay (EIA) was validated in detecting glucocorticoids in Pampas deer feces, in order to investigate the influence of several factors on the adrenocortical function. Fecal samples, behavioral data and information concerning male grouping and antlers status were collected at a monthly basis during a 1 year period from free-ranging stags living at Emas National Park, Brazil (18 degrees S/52 degrees W). The results revealed that concentrations of fecal glucocorticoids in winter were significantly higher than those corresponding to spring and summer. In addition, dry season data presented higher levels than during the wet season. Significant difference was found between fecal levels of breeding stags in summer and nonbreeding stags, whereas no difference was observed between breeding stags in winter and nonbreeding stags. On the other hand, males from areas with frequent human disturbance exhibited higher glucocorticoid concentrations and flight distances than individuals from areas of lower human activity. Males with antlers in velvet had elevated levels compared with animals in hard antler or antler casting. Also, we found that glucocorticoid levels were higher in groups with three or more males than in groups with only one male. The flight distances showed positive correlation with fecal glucocorticoid. These data indicate that fecal glucocorticoid provides a useful approach in the evaluation of physiological effects of environment, inter-individuals relationship and human-induced stressors on free-ranging Pampas deer stags.

Animals↗
PubMed2002

Dose confirmation studies of moxidectin 1% non-aqueous injectable and moxidectin 0.5% pour-on formulations against experimentally induced infections of larval and adult stage Oesophagostomum radiatum and Trichuris discolor in cattle.

Separate controlled trials were conducted to evaluate the efficacy of two formulations of moxidectin (1% non-aqueous injectable solution and 0.5% pour-on (Cydectin) against larval or adult stages of Oesophagostomum radiatum and Trichuris discolor infecting cattle. Fifty-three strongylate-free dairy breed steer calves were obtained from commercial sources. After a brief acclimation period, calves were randomly divided into two pools to evaluate the efficacy of the moxidectin formulations against targeted larval (n = 27 calves) or adult (n = 26 calves) parasites. Calves in the larvacidal trial were inoculated on Day -16 relative to treatment with approximately 1000 embryonated Trichuris spp. eggs and approximately 640 infective Oesophagostomum spp. larvae. Calves were allocated by lottery to one of three treatment groups (n = 8 per group), which included: Group 1--moxidectin 0.5% pour-on (0.5 mg/kg body weight (BW)) applied topically; Group 2--moxidectin 1% non-aqueous injectable (0.2 mg/kg BW) administered subcutaneously; Group 3--untreated controls. Treatments were administered on Day 0 and calves were housed by group with no contact among animals of different treatments. Three sentinel calves were necropsied on Day 0 of the larvacidal trial to assess viability of larval inocula. On Days 14, 15 and 16 after treatment, calves were euthanatized (two or three from each group per day) and samples of gut contents were collected for determination of total worm counts. On Day -63 relative to treatment, calves in the adulticidal efficacy trial were inoculated with approximately 1000 embryonated Trichuris eggs and then on Day -35 with approximately 2500 infective Oesophagostomum spp. larvae. Fecal samples were collected on Day -7 and the 24 calves with the highest egg counts were assigned by lottery to the following three treatment groups (n = 8 per group): Group 4--moxidectin 0.5% pour-on; Group 5--moxidectin 1% injectable; Group 6--untreated controls. Details of experimental treatments, calf housing and necropsy scheduling were similar to the larvacidal trial. In both the larvacidal and adulticidal trials, inocula contained a variety of parasites in addition to the targeted species. Based on geometric means, both moxidectin 0.5% pour-on and moxidectin 1% non-aqueous injectable significantly reduced (P < 0.05) numbers of Oesophagostomum spp. and Trichuris spp. with anthelmintic efficacies of >99% when used against adult or larval stages of infection. In addition, both formulations of moxidectin demonstrated >95% efficacy (P < 0.05) against larval stages of Strongyloides papillosus. The pour-on formulation had >97% adulticidal and larvacidal efficacy against Cooperia spp. females, while the injectable product was effective against female Cooperia spp. larvae and Cooperia oncophora adult males.

Administration, Topical↗
PubMedDate not supplied

Electropherotypic analysis of rotaviruses isolated from turkeys.

From 1980 to 1984, several flocks of turkeys in Minnesota exhibiting signs of clinical enteritis were examined for viruses. Electron microscopic (EM) examination of fecal specimens from 35 flocks revealed the presence of rotavirus particles. Rotaviruses were successfully isolated in cell cultures from only 24 of these positive fecal specimens. Double-stranded RNA (dsRNA) preparations made from these 24 cell-culture isolates and from the remaining 11 fecal samples that were rotavirus-positive on EM examination were analyzed by polyacrylamide gel electrophoresis for genetic differences in their genomes. The study revealed eight distinct electropherotypes among the rotavirus dsRNA preparations. Atypical dsRNA migration patterns were recognized only in preparations of dsRNA from fecal materials.

Animals↗
PubMed1988

Fecal alpha-1-antitrypsin excretion in acute diarrhea: relationship with causative pathogens.

Concentrations of alpha-1-antitrypsin (AT) in random fecal samples from 68 infants (92.7% outpatients) with acute diarrhea and 32 healthy controls were determined. The mean +/- s.d. for AT in infants with diarrhea was 2.07 +/- 1.88 mg/g dry stool (mg/g d.s.) compared with 1.29 +/- 0.72 mg/g d.s. (p less than 0.05) in controls. Fecal AT was significantly greater than that of the controls only for diarrhea caused by Rotavirus or Salmonella. Salmonella patients also had significantly higher fecal levels of AT than patients with diarrhea caused by Campylobacter. Fifty percent of the infants with Salmonella infections excreted more than 2.73 mg/g d.s. AT, which is above the control mean + 2 s.d. Fecal excretion of AT was highest in patients with macroscopic intestinal bleeding, but it still depended more on the causative agent than on bleeding per se. This finding of increased intestinal protein loss during acute diarrhea, even in the less severe diseases, emphasizes the necessity to rapidly restart adequate nutritional intake.

Acute Disease↗
PubMed2003

Detection of clarithromycin-resistant Helicobacter pylori in stool samples.

The recognition of the role of Helicobacter pylori in gastric diseases has led to the widespread use of antibiotics in the eradication of this pathogen. The most advocated therapy, triple therapy, often includes clarithromycin. It is well known that clarithromycin resistance is one of the major causes of eradication failure. The development of a rapid noninvasive technique that could easily be performed on fecal samples and that could also provide information about the antibiotic resistance of this microorganism is therefore advisable. Previous findings have demonstrated that clarithromycin resistance is due to a single point mutation in the 23S rRNA. All the mutations described have been associated with specific restriction sites, namely BsaI (A2143G), MboII (A2142C/G), and HhaI (T2717C). On this basis we have developed a new method, a seminested PCR, allowing screening for clarithromycin resistance of H. pylori directly on stool samples. This method furnished a 783-bp fragment of the 23S rRNA, which was subsequently digested by MboII, BsaI, and HhaI, in order to identify single point mutations associated with clarithromycin resistance. Of a total of 283 stool samples examined, 125 were H. pylori positive and two of them were shown to contain clarithromycin-resistant strains due to the presence of a mutation at position 2717, whereas no PCR products contained mutations at position 2142 or 2143. In order to evaluate the reliability of the new system, we compared the results of restriction analysis of the PCR products with the MICs shown by the H. pylori isolates by culturing gastric biopsies from the same patients.

Anti-Bacterial Agents↗
PubMed2005

An Austrian outbreak of Salmonella enteritidis phage type 36 in 2004.

In June 2004, an egg packing station was epidemiologically incriminated as possibly distributing the vehicle of the Salmonella enterica subsp. enterica ser. Typhimurium definitive type (DT) U291 outbreak which began in September 2003 and--as of June 2004 affected more than 100 persons all over Austria and 8 German tourists. A total of 68 egg producing farms were screened for salmonella. Whereas all samples tested negative for S. Typhimurium DT U291, one batch of laying hens was identified as being heavily contaminated with a rare Salmonella enterica subsp. enterica ser. Enteritidis phage type, namely PT 36. This phage type was cultured from 6 of 7 swabs from pooled fecal samples (drag swabs), from 1 of 4 samples of pooled yolks, and from 7 of 11 hens tested (ovaries, liver and intestinal content pooled per hen). In 2004, a total of 36 culture-confirmed human PT 36 infections (19 manifest cases, 12 asymptomatic carriers, and 5 excreters with unknown clinical status) were documented in 4 of the 9 Austrian provinces. It was possible to demonstrate that eggs from this single farm were the probable source of infection for 30 of 31 culture-confirmed human infections. Only in the case of one patient, who regularly purchased eggs at the local market, could no connection to the farm be proven due to inability to trace the market's egg source. The PT 36 outbreak ceased after elimination of the contaminated flock. This episode underlines the considerable potential of adequate epidemiological and microbiological investigations of food-borne outbreaks to improve prevention and control of communicable diseases. The source of the S. Typhimurium DT U291 outbreak--which initially prompted this investigation--is still not identified and accounts for more than 300 human cases until August 2005.

Animals↗
PubMed1998

Effect of dietary deoxycholic acid and cholesterol on fecal steroid concentration and its impact on the colonic crypt cell proliferation in azoxymethane-treated rats.

To elucidate the role of fecal steroids in the malignant tumor formation of colonic epithelial cells, we examined the effects of dietary deoxycholic acid (DCA) and cholesterol (CHL) on fecal steroid concentrations and their impact on colonic crypt cell proliferation. Twenty 5-week-old male Fischer 344 rats were provided with either a control semisynthetic diet or the same diet supplemented with 0.15% DCA and 1% CHL (steroid diet) over a 5-week period. The effects of these two diets were compared among rats that were either injected with azoxymethane (AOM), a known gastrointestinal carcinogen, or saline. In a 2 x 2 factorial design, rats fed each of these diets were given two weekly subcutaneous injections of either AOM (15 mg/kg b.w.) or saline at 6 and 7 weeks of age. At 9 weeks of age, fecal samples were obtained for analysis of bile acids, CHL and its bacterial metabolites of intestinal microflora. At 10 weeks of age, animals were sacrificed and colonic proliferation was assessed as vincristine-accumulated mitotic figures per crypt. Rats fed the steroid diet had significantly elevated fecal bile acid (5x, P < 0.001) and neutral steroid (10x, P < 0.01) levels when compared to those fed the control diet. AOM treatment did not appear to influence these levels. However, rats injected with AOM had a significant increase (P < 0.001) in their rate of colonic cell proliferation as compared to saline-injected control animals on both diets. Furthermore, rats fed the steroid diet had a significantly higher (P < 0.001) cell proliferation rate than animals fed the control diet. The effects of AOM treatment and the steroid diet on cell proliferation were additive. Our results demonstrate that high concentrations of neutral and acid steroids in the colonic lumen can enhance carcinogen-induced elevated cell proliferation and thus may play a key role in the etiology of colon cancer.

Animals↗