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Efficiency and sensitivity of techniques for recovering nematode eggs from bovine feces.

Haemonchus contortus eggs were extracted from sheep feces and known numbers were added to helminthologically sterile bovine feces to provide samples with seven, 30 and 60 eggs per gram (epg). At 60 epg, dilution techniques (modified Cornell-McMaster and modified McMaster) tended to overestimate the number of eggs and more eggs were recovered (mean of 121 and 88% respectively) with these techniques than with centrifugal concentration procedures (modified Cornell-63% and Wisconsin- 69%). At 30 epg, all techniques were comparable (modified Cornell-McMaster 67%, modified McMaster 63%, modified Cornell and Wisconsin 64%). At 7 epg, the Wisconsin (61%), modified Cornell (60%) and Cornell-McMaster (94%) techniques were comparable and better than the modified McMaster technique (16%). At all levels of epg, the modified Cornell and Wisconsin techniques recovered eggs from 100% of the samples. The Cornell-McMaster and modified McMaster techniques recovered eggs from 90 and 100% of samples at 60 epg; 40 and 100% at 30 epg; and 21 and 11% at 7 epg. With a gravitational concentration procedure, the Standard Vial, no more than 16% of the eggs at any level of epg were recovered and at 7 epg eggs were recovered from only one-half of the samples. Five gravitational concentration techniques were assessed over 66 to 490 epg. The Ovassay, Fecalyzer and modified Standard Vial techniques were comparable in efficiency (28%, 25% and 24% respectively), but the Standard Vial technique was less efficient (11%). Introduced into diagnostic parasitology was the concept of predictive values which is the proportion of samples that a technique correctly identifies as being negative for parasite eggs. At 7 epg this was calculated to be zero for the modified Cornell-McMaster, modified McMaster and Standard Vial techniques and 100 for the Wisconsin and modified Cornell techniques.

Animals↗

[Desensitization immunotherapy on patients with mite-positive bronchial asthma using purified mite feces antigen fractions].

At present, we are performing desensitization immunotherapy on patients with mite-positive bronchial asthma using purified mite feces antigen fractions. We obtained the following results in 13 patients 4 to 12 months after the start of treatment. 1) The fraction showing the maximum reaction in the skin test was HM1-2 (molecular weight: 150-155 kD) in two patients, HM2 (30-40 kD) in nine patients and HM3 (10-20 kD) in two patients, and immunotherapy was performed using these fractions. None of the patients showed the maximum skin reactions or was treated with HM4 (less than 10 kD). 2) In nine patients with perennial asthma, the effects of treatment were excellent in four and moderate in three, while two cases remained unchanged. In four patients with seasonal asthma, the treatment was effective in all cases. 3) In patients in whom immunotherapy was effective, specific IgG antibody increased after the treatment, but it did not increase in the unchanged cases. Among the subclasses of specific IgG antibody increased by the treatment, no clear changes were seen in IgG4 antibody and no conclusion has been reached at present. 4) None of the patients had definite adverse reactions such as the occurrence of asthma attacks or anaphylaxis. These results suggested that purified mite feces antigens are safe and effective. We plan to continue this study with more patients in the future.

Adolescent↗

High-molecular-size mite allergens with high carbohydrate content in mite extract containing mite feces of Dermatophagoides farinae.

A series of mite allergens with high carbohydrate content was fractionated from mite extract containing mite feces by ultrafiltration, and successive chromatography on Ultrogel AcA 54, Sepharose CL-6B, and DEAE Toyopearl 650S. The final fractions isolated were termed HM1-2-Eff, -0.1A, -0.1B, -0.3A, -0.3B, and -1M according to the eluted salt concentration. These six fractions were stained weakly with Coomassie Brilliant Blue but strongly with periodate-Schiff's reagent on SDS-PAGE. Their molecular weight were estimated to be 150-155 KD on Sepharose CL-6B. All of these six fractions provoked strong erythema in skin of mite-sensitive and asthmatic patients. HM1-2-Eff, -0.1A, -0.1B and -0.3B reacted with 73% of mite-sensitive patients' IgE by the nitrocellulose dot blotting. One ng/ml of 4 fractions (HM1-2-0.1A, 0.3A, 0.3B and -1M) released histamine from mite allergic patient's blood cells. HM1-2-0.3A induced proliferation of T cell from immunized mouse by mite extract containing mite feces (Dff).

Allergens↗

[Glycoprotein hexoses in feces of infants with lactose intolerance].

A modified method for estimation of total glycoprotein hexoses in feces, based on their measurements in the blood serum, is presented. Sixty-six nursing children with lactose intolerance, breastfed or formula fed, were examined; formula fed babies were kept on mixtures with high and low lactose content. Glycoprotein hexose parameters were as follows (X +/- m): 13.51 +/- 1.93, 12.05 +/- 2.20, and 3.69 +/- 0.47 g/l feces. In control children without lactose intolerance (n = 33) this value was 3.6 +/- 0.79 g/l. Increased glycoprotein excretion is connected with glycocalix and small intestinal enterocyte alteration.

Breast Feeding↗

The carriage of Escherichia coli on the peri-urethral area and in the feces in patients on long term low dose cotrimoxazole therapy.

Peri-urethral and fecal carriage of E. coli has been studied in 30 patients maintained on long term low dose cotrimoxazole and in abacteriuric subjects not receiving any antibiotic. Cotrimoxazole treatment is associated with a dramatic reduction in E. coli carriage both in the feces and especially on the peri-urethral area which persists over months or years. Recovery of trimethoprim-resistant E. coli from the feces was uncommon but when observed antedated subsequent relapse due to resistant organisms. Possible mechanisms for low peri-urethral carriage rates include reduction in fecal carriage, vaginal secretion of trimethoprim or low urinary concentrations of that drug.

Adult↗

Short-chain carboxylic acid in the feces in patients with pancreatic insufficiency.

Short-chain carboxylic acids in the feces of 12 patients with chronic pancreatitis having steatorrhea and 18 healthy subjects were determined by high-performance liquid chromatography. The fecal amount of the patients with chronic pancreatitis was increased to about twice that of the control, and the fat in the feces increased to eight times. The excretion of short-chain carboxylic acids increased to about three times. The short-chain carboxylic acid concentration regarded as derived from protein or amino acid, such as iso-butyrate, isovalerate, and n-valerate, was significantly increased in patients with chronic pancreatitis. From the above results, our data suggest that there is maldigestion of protein besides that of carbohydrate. And we confirmed that these data are an in-vivo illustration of the phenomena which have been studied in-vitro experiments, when fecal bacteria were incubated with peptide or amino acids as a substrate.

Adult↗

Use of metabolic inhibitors to study H2 consumption by human feces: evidence for a pathway other than methanogenesis and sulfate reduction.

Human fecal bacteria produce and consume hydrogen, and consumption rate is a critical determinant of the volume of H2 liberated into the colonic lumen. Two bacterial reactions that have been reported to be major consumers of H2 are methanogenesis and sulfate reduction. To determine the importance of each of these reactions, we measured the disappearance of exogenous H2 during incubation with human fecal homogenates treated with 20 mmol/L 2-bromoethanesulphonic acid (BES), an inhibitor of methanogenesis, and/or 20 mmol/L sodium molybdate (Mo), an inhibitor of sulfate reduction. Four methanogenic and four nonmethanogenic samples consumed an average of 99% and 85%, respectively, of the initial H2 during 22 hours of incubation. With methanogenic homogenates, 36% of the H2 consumption persisted despite inhibition of methanogenesis. Inhibition of sulfate reduction had no effect on the rate of H2 consumption by these eight fecal specimens. The importance of fecal sulfate availability was determined in fecal samples obtained from an additional 14 randomly selected volunteers. Incubation after supplementation with 20 mmol/L sulfate was associated with an increase in sulfide production in four of the samples, and three of these four samples also demonstrated an increased rate of H2 consumption. No such evidence of sulfate reduction was observed in the other 10 homogenates. We conclude that a bacterial reaction other than methanogenesis and sulfate reduction, perhaps the recently described reduction of CO2 to acetate, represents a major metabolic route of H2 disposal in nonmethanogenic feces and a minor, but appreciable, pathway in methanogenic feces.

Feces↗

[Lysozyme activity in cat feces].

Activity of fecal lysozyme was determined in healthy cats and cats with chronic diarrhea. The established reference value ranged from 0 up to 2.6 micrograms/g feces. Mean activity of the control group was 1.3 +/- 0.9 micrograms/g feces, mean activity of the patient group was 3.4 +/- 3.3 micrograms/g. There was a statistically significant difference between these two values. Nevertheless it was not possible to classify various diseases by evaluating lysozyme activity. In addition to endoscopy and histological examination, determination of lysozyme can be used as screening test and means of assessment for treatment and progress in cats with chronic diarrhea.

Animals↗

[Demonstration of activity of two potentiated sulfonamides in feces of horses after oral or intravenous administration].

Both, the oral and intravenous application of two trimethoprim-potentiated sulfonamides induced measurable antibacterial activities in the feces of horses. With regard to the risk of antibiotic-induced alterations of the gastrointestinal flora, the route of application of potentiated sulfonamides seems to be of minor importance. The antibiotics used were Sulfadimethoxine/Trimethoprim (Trafigal 30% ad us. vet.) for oral and Sulfadoxine/Trimethoprim (Borgal 24% ad us. vet., both Hoechst AG, Frankfurt) for intravenous application. As recommended, both drugs were given in a dose of 20 mg per kg bodyweight. The detection method is based on a procedure layed down in German laws for sulfonamide residues in meat-samples and has undergone some modifications for the examination of feces.

Administration, Oral↗

Isolation, purification and biological activity of major docetaxel metabolites from human feces.

We have developed a procedure suited for the isolation of metabolites of docetaxel (Taxotere) from human feces. The compounds were extracted from the feces with diethyl ether and further purified by (semipreparative) HPLC. Four metabolic products were obtained in submilligram quantities. Analytical HPLC with photodiode array detection showed that the purity of each compound was higher than 98%. There structures have been characterized by UV absorption and FAB/MS. All four compounds were oxidation products of the tert-butyl group attached to the C13-side chain, and corresponded to structures identified previously. The purified products were used for evaluating their cytotoxic activities against a human ovarian cancer (A2780) and a rat colon cancer (CC531) cell line, and their myelosuppressive effects in a hematopoietic progenitor toxicity assay. Although distinctions in biological activities between the compounds were evident, all metabolites showed a marked reduction in both cytotoxic and myelotoxic properties.

Aged↗

Assessment of pregnancy in Kiang mares (Equus hemionus holdereri ) using estrogen determination in feces.

Analysis of fecal estrogens was used to diagnose pregnancy in 6 Kiang mares (Equus hemionus holdereri ) that were kept at Tierpark Berlin. Three extraction methods were compared and were followed by an established RIA for total estrogen. Extraction of desiccated feces with chloroform/n-hexane and KOH, with and without enzyme hydrolysis showed better results than extraction with diethylether without hydrolysis. Pregnancy was confirmed by observation of foaling in 2 mares that showed estrogen concentrations between 800 and 1800 ng/g and in 1 mare that showed widely fluctuating values between 500 and 1300 ng/g of feces. Two mares with estrogen concentrations below 500 ng/g were not seen to foal. The method using chloroform/n-hexane and KOH without enzyme hydrolysis seems practical for non-invasive evaluation of the endocrine status in this endangered Equidae species.

Journal Article↗

Direct ELISA for estrone measurement in the feces of sows: prospects for rapid, sow-side pregnancy diagnosis.

Both free and conjugated fecal estrogens were surveyed in sows by means of RIA's after extraction and column chromatography. Six different RIA's were performed using the same fecal suspension. Based on differences in concentration in feces from 6 pregnant and 4 nonpregnant sows, estrone (E1) was selected for the development of a homologous, competitive ELISA for pregnancy diagnosis purposes. Polyclonal antibodies were raised against 6-keto-E1-carboxymethyloxime coupled to BSA. Biotinylated E1 was used as the competitive agent and was detected with a streptavidin-peroxidase conjugate. Tetramethylbenzidine was used as chromogen. Validation of this direct, easy to perform ELISA showed satisfactory specificity, accuracy, precision and sensitivity. Comparison of the assay with a validated RIA (including extraction and column chromatography) resulted in a linear regression equation of ELISA = 0.88 RIA - 0.72 (r = 0.95; n = 24 fecal samples). The E1 values in feces from 11 pregnant sows during the first 40 d of gestation compared with values from 10 nonpregnant sows showed significant differences between Days 23 and 35 of pregnancy (P < 0.05). The highly significant difference in E1 concentrations (P < 0.0012) between Days 27 and 30 in particular offers promise for pregnancy diagnosis.

Journal Article↗

8-Azido-ATP labelling of the FecE protein of the Escherichia coli iron citrate transport system.

The iron(III) transport system via citrate displays the characteristics of a binding protein-dependent transport mechanism through the cytoplasmic membrane. One of the five transport proteins, FecE, contains the two motifs found in ATP-binding proteins. Cloned overexpressed FecE was photoaffinity-labelled with [32P]8-azido-ATP. Labelling was inhibited by 1 mM ATP, ADP, GTP and less by CTP, demonstrating the specificity of the reaction. It is suggested that ATP is the principal energy source for iron(III) citrate transport through the cytoplasmic membrane of Escherichia coli.

Adenosine Triphosphate↗

Anaerofustis stercorihominis gen. nov., sp. nov., from human feces.

Phenotypic and phylogenetic studies were performed on an unidentified Gram-positive, strictly anaerobic, non-spore-forming, rod-shaped bacterium isolated from human feces. The organism was catalase-negative, resistant to 20% bile, produced acetic and butyric acids as end products of glucose metabolism, and possessed a G+C content of approximately 70 mol%. Comparative 16S rRNA gene sequencing demonstrated that the unidentified bacterium was a member of the Clostridium sub-phylum of the Gram-positive bacteria, and formed a loose association with rRNA cluster XV. Sequence divergence values of 12% or greater were observed between the unidentified bacterium and all other recognized species within this and related rRNA clusters. Treeing analysis showed the unknown anaerobe formed a deep line branching near to the base of rRNA cluster XV and phylogenetically represents a hitherto unknown taxon, distinct from Acetobacterium, Eubacterium sensu stricto, Pseudoramibacter and other related organisms. Based on both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium from feces be classified in a new genus Anaerofustis, as Anaerofustis stercorihominis sp. nov. The type strain of Anaerofustis stercorihominis is ATCC BAA-858(T)=CCUG 47767(T).

Journal Article↗

Subdoligranulum variabile gen. nov., sp. nov. from human feces.

During studies on the microflora of human feces we have isolated a strictly anaerobic, non-spore-forming, Gram-negative staining organism which exhibits a somewhat variable coccus-shaped morphology. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism is phylogenetically a member of the Clostridium leptum supra-generic rRNA cluster and displays a close affinity to some rDNA clones derived from human and pig feces. The nearest named relatives of the unidentified isolate corresponded to Faecalibacterium prausnitzii (formerly Fusobacterium prausnitzii) displaying a 16S rRNA sequence divergence of approximately 9%, with Anaerofilum agile and A. pentosovorans the next closest relatives of the unidentified bacterium (sequence divergence approximately 10%). Based on phenotypic and phylogenetic considerations, it is proposed that the unusual coccoid-shaped organism be classified as a new genus and species, Subdoligranulum variabile. The type strain of S. variabile is BI 114(T) (=CCUG 47106(T)=DSM 15176(T)).

Journal Article↗

Allobaculum stercoricanis gen. nov., sp. nov., isolated from canine feces.

Morphological, biochemical, and molecular genetic studies were performed on an unknown anaerobic, catalase-negative, non-spore-forming, rod-shaped bacterium isolated from dog feces. The unknown bacterium was tentatively identified as a Eubacterium species, based on cellular morphological and biochemical tests. 16S rRNA gene sequencing studies, however, revealed that it was phylogenetically distant from Eubacterium limosum, the type species of the genus Eubacterium. Phylogenetically, the unknown species forms a hitherto unknown sub-line proximal to the base of a cluster of organisms (designated rRNA cluster XVI), which includes Clostridium innocuum, Streptococcus pleomorphus, and some Eubacterium species. Based on both phenotypic and phylogenetic criteria, it is proposed that the unknown bacterium be classified as a new genus and species, Allobaculum stercoricanis. Using a specific rRNA-targeted probe designed to identify Allobaculum stercoricanis, in situ hybridisation showed this novel species represents a significant organism in canine feces comprising between 0.1% and 3.7% of total cells stained with DAPI (21 dog fecal samples). The type strain of Allobaculum stercoricanis is DSM 13633(T)=CCUG 45212(T).

Journal Article↗

Evaluation of false ultrasonographic pregnancy diagnoses in sows by measuring the concentration of unconjugated estrogens in feces.

On Days 26, 28, and 30 after AI, ultrasonographic pregnancy diagnoses were performed on 207 gilts and sows by using a 3.5 MHz linear-array transducer. Fecal samples were taken from the rectum after each ultrasonographic examination, and the concentrations of unconjugated estrogens in selected samples (n = 73) were measured by RIA. Fecal unconjugated estrogen concentration of 11.7 ng/g feces or higher was indicative of pregnancy. The overall sensitivity and specificity of the ultrasonographic test was 99% for farrowing sows and 73.1% for nonfarrowing sows. With one exception, sows with a false negative diagnosis by ultrasonography on Day 26 were correctly diagnosed pregnant by elevated fecal unconjugated estrogens or repeated ultrasonographic examinations on Days 28 or 30. Return to estrus around the sampling period may cause false positive results in the unconjugated estrogen assay, while early embryonic mortality can result in false positive diagnoses in both the ultrasonographic test and estrogen assay. Although there was a positive correlation between the concentrations of unconjugated estrogens in the feces and litter size at farrowing in the selected sows, it seems very unlikely that fecal estrogens can provide an accurate tool for predicting litter size.

Journal Article↗

Field studies on inhibitory influence of skin pressure exerted by a body compensatory brassiere on the amount of feces.

The present experiment investigated the effects of skin pressure produced by a body compensatory brassiere on defecation activity. Seven healthy females (11-41 yrs) volunteered as participants, being free of medication and constipation. The experiment lasted 3 weeks. The participants did not wear the body compensatory brassiere for the first week, wore it during waking hours for the second week, and again did not wear it for the third week. Whenever they desired to defecate, they did so and then weighted the amount of feces immediately by themselves. Eating times, daily amounts of foods and drinks, their menu, work intensity and its duration, retiring and rising time were controlled to be as similar as possible from day to day. The main finding was that the amount of feces was significantly smaller during the second week (wearing the body compensatory brassiere) than the first and third weeks (not wearing the body compensatory brassiere). These observations are discussed in terms of the suppression of the parasympathetic nervous system and intestine motility, and the delayed transit time in the large intestine.

Adolescent↗