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Indirect parameters of pancreatic function in cystic fibrosis (CF) during a controlled double-blind trial of pancreatic supplementation.

During two treatment periods (4 weeks each), serum immunoreactive trypsin (IRT), immunoreactive human lipase in stool (IRL), and chymotrypsin (CT) activity in stool were determined in 16 cystic fibrosis patients and compared with fecal fat excretion (72-h sampling). Fecal fat estimation revealed mild to severe steatorrhea in all 16 patients (X = 13.7 +/- 9.0 g/24 h) in at least one study period. Stool fat excretion was highest in underweight adolescents and adults. Comparison of IRT and IRL with stool fat values showed no significant statistical correlation. IRT values revealed an inverse exponential correlation with age, with a steep decline at the age of 5 years. CT levels were very high in 14 of our 16 patients during supplementation therapy, whereas 2 patients showed subnormal CT values. We conclude that since indirect parameters of pancreatic function do not correlate with stool fat excretion, stool fat remains the best indirect parameter for the assessment of pancreatic insufficiency in cystic fibrosis. Leaving pancreatic enzyme supplementation in cystic fibrosis patients on the basis of normal serum trypsin or fecal lipase values does not appear to be adequate.

Adolescent↗

[Polymerase chain reaction (PCR) method for the detection of Salmonella by using Chelex resin after enrichment culture].

Salmonella has been implicated as a important pathogen associated with acute gastroenteritis or food borne diseases in humans. Rapid and accurate diagnostic tests for Salmonella are needed by both the food industry and clinical laboratories. A pair of oligonucleotide primers from the phoE gene was used to develop a polymerase chain reaction (PCR) procedure to detect Salmonella spp. When Chelex-extracted DNA from the SBG enrichment culture were used as templates for the PCR amplification (E-Chx), the positive results were obtained with a few cells of Salmonella. This highly specific and sensitive technique gave an efficiency of 1.2 times (73 samples were positive) when tested against conventional culture method (62 samples were positive) using 125 fecal samples stored at 4 degrees C for 1-16 months. Based on the results obtained in this study, it appears that the enrichment-PCR method (E-Chx) using Chelex-extracted DNA described here can be useful to screen a large number of samples such as foods or environmental specimens for contamination of Salmonella.

Base Sequence↗

Effect of cheese on intestinal transit time and other indicators of bowel function in residents of a retirement home.

Effects of increased intake of cheese on intestinal transit time and other indicators of bowel function were studied in 21 retirement home residents (18 women and 3 men; age, 68-87 years). The study was divided into four succeeding periods: 1) 1-week basal period (usual diet); 2)3-week cheese period (extra cheese was offered as such on bread or used in cooking); 3) 3-week no-cheese period (all cheese on bread was replaced with cured meats and cold cuts, and no cheese was used in cooking); 4) 3-week follow-up period (usual diet). During the last week of each period a questionnaire was filled out on fecal frequencies, consistency of feces (soft, normal, hard), and occurrence of abdominal pain and flatulence. Use of laxative medications and therapeutic foods (prunes) was registered. Eleven of the 21 subjects collected fecal samples for the determination of fecal wet weight and intestinal transit time by means of radiopaque Sitzmark capsules. Intakes of cheese, fiber-containing foods, and fluids by these 11 subjects during meals offered in the cafeteria were recorded on a prefilled questionnaire. In spite of a 10-fold increase in the intake of cheese no change in intestinal transit time, fecal frequency, fecal wet weight, consistency of feces, and occurrence of gastrointestinal symptoms was observed. The use of laxative medication was higher during the cheese period, but no change in the combined use of laxative medication and therapeutic foods (prunes) was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Influence of dietary calcium and protein on fecal pH, consistency, and rate of passage in dairy cattle.

Sixteen Holstein heifers and 20 calves were in trials on the effect of 10 and 15% crude protein and .3 and .9% calcium in the diet on fecal pH and consistency and on rate of passage of the liquid phase of digesta. Time of first appearance of polyethylene glycol was an indicator of rate of passage of the liquid phase in the lower tract of the animal. Respective fecal pH and standard errors for heifers and calves increased from 5.79 +/- .03 and 5.78 +/- .03 on low calcium diets to 6.24 +/- .03 and 6.33 +/- .03 on high calcium diets. High protein diets increased fecal pH from 5.89 +/- .03 and 5.97 +/- .03 to 6.13 +/- .03 and 6.14 +/- .03 for heifers and calves. Effects of calcium and protein were additive. Neither dietary protein nor calcium affected consistency of feces or first appearance of polyethylene glycol in feces. Mean first appearances of PEG in feces were 683 +/- 13 min for heifers and 528 +/- 18 min for calves. Small correlations between fecal pH and fecal consistency and between fecal pH and first appearance of polyethylene glycol in feces indicate little or no relationship of fecal pH to these two variables. From variance in the heifer trial, 10 fecal samples are required to estimate fecal pH within +/-.16 pH units with 95% confidence.

Animals↗

Detection of Shigella dysenteriae type 1 and Shigella flexneri in feces by immunomagnetic isolation and polymerase chain reaction.

A combination of immunomagnetic separation (IMS) and a polymerase chain reaction (PCR) procedure was used for direct isolation and identification of Shigella dysenteriae type 1 and Shigella flexneri from feces. Immunomagnetic particles were coated with monoclonal antibody MASFB, which is specific for a common epitope of the O polysaccharides of S. dysenteriae type 1 and S. flexneri. Bacteria bound to the beads were boiled in water, and target DNA was amplified with a primer pair specific for a gene coded on the invasion-associated locus (ial) of the large virulence plasmid of all four Shigella spp. and enteroinvasive strains of Escherichia coli. A 320-bp DNA fragment was generated and detected by an alkaline phosphatase-conjugated probe. Nonviable cells were also captured and detected by this technique. The method is simple and fast (7 h) and has a detection limit of ca. 10 Shigella organisms per g in fecal samples. The combined IMS-PCR assay correctly identified all 57 samples carrying S. dysenteriae type 1 and 68 samples carrying S. flexneri from 238 fecal specimens and also permitted detection of 17 samples carrying Shigella spp. from 113 specimens from diarrheal patients in whom shigellae were not detected by conventional culture.

Antibodies, Monoclonal↗

Rhodococcus equi in fecal and environmental samples from Kansas horse farms.

Horse farms in Kansas were surveyed for the incidence of Rhodococcus equi. Fecal specimens and soil or cobweb samples were collected from each farm and cultured on selective media. One control farm (with no history of R. equi infection), one farm which had an outbreak 3 and 4 years previously and 2 farms which had R. equi-infected foals that season were surveyed. In addition, fecal samples from 21 horses hospitalized in the Kansas State University Veterinary Hospital were cultured. There was no significant difference in the incidence of R. equi in fecal samples from the 2 farms with recent disease problems. The farm with a history of disease had a significantly higher percentage of positive fecal cultures than the 2 farms with a more recent history of disease. Neither ration composition nor sex of the horses appeared to affect the fecal culture results. Fecal samples from 2 birds on 1 farm were positive for R. equi.

Animals↗

The effect of toilet sanitizers and detergents on immunological occult blood tests.

Effects of time-releasing toilet bowl sanitizers and their main constituents, detergents, on immunological fecal occult blood tests were examined. Six kinds of toilet sanitizers and 19 kinds of detergents were included. Patients' fecal samples and hemoglobin solutions were measured by means of quantitative immunological fecal occult blood tests after being mixed with sanitizer or detergent solutions. Marked reduction in hemoglobin concentration or antigenicity was observed after the addition of sanitizers or detergents at a final concentration of 0.5 g/l. Most of these inhibitory reactions were dose dependent. In contrast with the immunological method, the chemical method using o-tolidine and guaiac were not affected by the detergents tested. In sampling feces for immunological fecal occult blood tests, contact of samples with sanitizer solutions should be avoided.

Detergents↗

Bifidobacterium species isolated from animal feces and from beef and pork meat.

Bifidobacteria were isolated from 122 of 145 samples of animal feces (from cattle, swine, sheep, goats, horses, rabbits, chickens, geese, and pigeons) from farms in France and Austria and from 92 of 955 production and processing chain samples of beef and pork (obtained at slaughter, cutting, and retail). Bacterial strains were identified to species by phenotypic numerical classification based on API 50CH and ID 32A tests and DNA-DNA hybridization. Bifidobacterium pseudolongum was present in 81% (99 of 122 samples) of all Bifidobacterium-positive fecal samples and predominated in samples from all animal species except those from swine from Austria. In these Austrian swine samples, the majority of strains were identified as Bifidobacterium thermophilum (78%), followed by B. pseudolongum (48%). The distribution of B. thermophilum and B. pseudolongum differed significantly between Austrian swine and cattle samples such as those collected along beef and pork production and processing chains. Bifidobacterium animalis was isolated from swine feces, and Bifidobacterium ruminantium was isolated from cow dung. Six fecal isolates (from cattle, swine, rabbits, goats, and horses) were identified as belonging to Bifidobacterium species of predominantly human origin: B. adolescentis, B. bifidum, and B. catenulatum. Only one other species, Bifidobacterium choerinum, was detected with low frequency in a pork processing chain. B. pseudolongum subsp. pseudolongum was predominant in pig feces, whereas B. pseudolongum subsp. globosum was predominant in feces from other animal species. Four strains closely related to both subspecies (58 to 61% DNA reassociation) formed a distinct genomic group. PCR techniques, which are more rapid and sensitive than culture-based methods, could be used to detect directly B. pseudolongum and B. thermophilum as indicators of fecal contamination along the meat processing chain.

Animals↗

A survey of Blastocystis sp. in livestock, pets, and zoo animals in Japan.

The prevalence of Blastocystis sp. was examined in fecal samples collected from cattle, pigs, dogs, and a variety of zoo animals (primates, carnivores, herbivores, pheasants, and ducks) by direct observation of fresh fecal suspensions or cultured materials, using light microscopy. The cattle and pigs were randomly sampled from 11 and 12 commercial farms, respectively, located in the western region of Japan. The dog material used in this study was obtained from pets housed in an animal shelter in the city of Osaka. Zoo animals were chosen based on housing conditions that minimized the possibility of intra-zoo transmission of the organism. The prevalence rate among the groups varied greatly. A high prevalence of infection was observed in the farm animal group, ranging from 95% (58/61) in pigs to 71% (39/55) in cattle, whereas the dog fecal samples were completely free of the organism. Prevalence of the organism in the zoo animal were 85% (29/34) in primates, 80% (8/10) in pheasants, 56% (9/16) in ducks, and 0% (0/58) in various carnivores and herbivores. Among the zoo animals infected with Blastocystis, eight species of primates, eight species of pheasants, and four species of ducks were confirmed as new hosts. Since Blastocystis organisms isolated from various animals were morphologically indistinguishable from Blastocystis hominis by light microscopy, further genomic studies are required for analysis of the zoonotic potential or etiological significance of these isolates.

Animals↗

Phosphorus concentration and solubility in dairy feces: variability and affecting factors.

Recent data from phosphorus (P) feeding trials have demonstrated that P concentration in dairy feces is directly affected by P levels in diets and that farm P surpluses as well as potential environmental losses can be reduced through dietary manipulation. The current study was conducted to examine the variability of fecal P under farm conditions and to elucidate factors affecting the concentration and solubility of fecal P. Feed and fecal samples from >30 commercial dairies in the Northeast and Mid-Atlantic regions were analyzed. Dietary P concentrations ranged from 3.45 to 5.78 g/kg of feed DM (DM), and P determined in acid digests (TP) of feces from 5.84 to 12.84 g/kg of fecal DM. On average, 50% of fecal TP was water soluble; of the latter, 83% was inorganic (Pi). Across-farm variability (n=33) had CV averaging 18.9% for fecal TP and >20% for Pi and total P (Pt) in water extracts. Within-farm variability based on multiple samples per herd had the same magnitude as across-farm and was independent of sample numbers from individual farms (n=7 to 30). Of all fecal parameters determined, pH and DM had the lowest variability (CV <10%), water-soluble Pi, Pt, and Ca the highest (CV of 20 to 30%), and total P, Ca, and Mg determined by acid digests were intermediate (CV 10 to 20%). Water-soluble Pi concentrations determined in dried-ground fecal samples were lower than in wet samples. The drying-grinding process changes Pi solubility and the change is not linear. This study confirms that dietary P concentration is the dominating factor affecting fecal P excretion; however, Ca concentration, DIM, and fecal pH also made small, but statistically significant contributions, although some of the mechanisms remain to be thoroughly investigated.

Animal Feed↗

[Evaluation on the dietary iron absorption by young Tibetan men using stable isotopic tracer].

OBJECTIVE: The study was aimed to evaluate the iron absorption in the Tibetan man's diet using the stable isotopic iron, and present a reference to improve the dietary iron absorption for Chinese. METHODS: In the study conducted on the spot of high altitude (> 3500m), the iron tracer (FeSO4 enriched with 57Fe) and the recovery indicator dysprosium (Dy) were orally administrated to 16 young Tibetan men, and their feces were sampled. The ratios of 57Fe/56Fe in fecal samples was determined by inductively coupled plasma mass spectrometry, and then the iron absorption was calculated based on the principle of isotope dilution. RESULTS: The rate of dietary iron absorption in young Tibetan men was 13.4% +/- 6.4% with good linear correlation between daily fecal excretions of iron tracer and Dy. CONCLUSION: Dy had the same behavior as non-absorbed iron tracer in the digestive tract and the recovery of Dy may indicate the completeness of fecal collection. Dietary iron absorption in the adult Tibetan man was relatively high.

Adolescent↗

Effect of eprinomectin treatment at calving on milk production in dairy herds with limited outdoor exposure.

The objective of this study was to determine the effect of anthelmintic treatment at calving in herds that were totally or semiconfined during the summer. In totally confined herds, lactating and dry cows were housed throughout the summer and had no access to pasture. In semiconfined herds, lactating and dry cows had limited outdoor exposure to a small pasture or paddock but were still fed a ration that met all their nutritional requirements. The study was carried out between February 2002 and February 2003 in 65 herds enrolled with DHI and distributed in 4 regions in Canada and 1 state in the United States. Cows were randomly allocated to receive eprinomectin or a placebo, with treatment being administered on or close to day of calving. In May and June 2002, 8 fecal samples were collected from each farm and fecal egg counts (FEC) were determined. Monthly bulk tank milk samples from each farm were tested with an indirect ELISA using a crude Ostertagia ostertagi antigen. Monthly test-day milk production data were recorded for 200 d after calving. In general, FEC were very low (mean = 1 egg per gram, range = 0 to 27). Mean herd bulk milk ELISA optical density ratio (ODR) values for the whole year ranged between 0.22 and 0.80. The ODR values were dichotomized into high and low using a threshold of 0.5. Treatment effects were analyzed using a linear mixed model with herd and cow as random effects. The analysis was restricted to 4789 cows (23,956 test-day records) treated between 21 d before and 7 d after calving. Overall, there was no significant effect of treatment. However, there was a tendency for an interaction between treatment and ODR, as illustrated by a larger numerical difference in treated vs. untreated cows in high-ODR herds than in low-ODR herds. However, the confidence intervals for the treatment effects (kg/d of milk per cow) in high-ODR herds (-0.33 to 1.10) and in low-ODR herds (-0.53 to 0.14) were wide and included zero. Therefore, this study failed to show a beneficial effect of eprinomectin treatment in these totally or semiconfined herds.

Animal Husbandry↗

Effects of antiinflammatory agents on chronic Salmonella typhimurium infection in a mouse model.

Anti-inflammatory corticosteroids have been demonstrated to lower the resistance of the host to a range of infections in several different animal models. We have adapted the mouse model for Salmonella typhimurium infection, using a subcutaneous inoculation of 10(4) organisms in CBA/Ca mice. The chronic infection was potentiated by treatment with hydrocortisone given orally in the diet for up to 4 weeks, beginning on day 30 after infection. A dose of 25 mg/kg of body weight resulted in 100% mortality from Salmonella infection within 24 days of beginning hydrocortisone treatment. Nonsteroid drugs, indomethacin and a trial agent BW 755C, were compared with the corticosteroid in this model. No mortality was recorded during the 4 weeks of drug treatment in these groups. Fecal samples were monitored weekly, and drug-related effects on the normal fecal flora were observed. The kinetics of S. typhimurium infection were followed in both fecal and organ samples for the duration of the experiments. The model is useful for the determination of the lowered resistance of the host to infection after drug treatment and specifically demonstrated here the potential of the trial drug BW 755C as an active anti-inflammatory agent without one of the undesirable side effects of corticosteroids.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗

Evaluation of a test for Clostridium difficile toxins A and B for the diagnosis of neonatal swine enteritis.

A commercially available 1-hour enzyme immunoassay (EIA) for detecting the presence of Clostridium difficile toxins A and B was evaluated for use in diagnosis of C. difficile infections in neonatal swine. This test was compared with a tissue culture cytotoxicity assay, which is considered to be the reference standard for the detection of C. difficile toxins. Twenty-seven samples of colonic contents and 23 fecal samples were collected from freshly euthanized neonatal swine with a history of scours. Of the 50 specimens tested, 20 were positive by the EIA test and tissue culture and 24 were negative by both tests, for an overall correlation of 88%. The sensitivity and specificity of the EIA were 91% and 86%, respectively, and the positive and negative predictive values were 84% and 86%, respectively. The EIA test is considered suitable as an aid for the diagnosis of C. difficile enteritis because of the high correlation between EIA results and those of the tissue culture cytotoxicity assay.

Animals↗

Development of an antibody-based capture enzyme-linked immunosorbent assay for detecting Echinostoma caproni (Trematoda) in experimentally infected rats: kinetics of coproantigen excretion.

The present study reports on the development of a coproantigen capture enzyme-linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory-secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49-56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections.

Animals↗

Antimicrobial susceptibility patterns of Salmonella isolates from cattle in feedlots.

OBJECTIVE: To evaluate the antimicrobial susceptibility patterns of Salmonella isolates from feedlot cattle. DESIGN: Cross-sectional study. SAMPLE POPULATION: 263 Salmonella isolates. PROCEDURES: Fecal samples were collected from the floor of 2 pens in each of 100 feedlots. Two hundred eighty Salmonella isolates were recovered after bacteriologic culture from 38 pens. Of these, 263 isolates were available for antimicrobial susceptibility testing to 16 antimicrobials, using microbroth dilution breakpoint plates. RESULTS: Less than 5% of isolates were resistant to any of the antimicrobials tested, with the exception of sulfamethoxazole (15; 5.7%) and tetracycline (61; 23.2%). Most isolates (197; 74.9%) were susceptible to all antimicrobials tested, whereas 18 (6.8%) were resistant to 2 or more antimicrobials. The percentage of isolates with resistance to any antimicrobial varied by serotype. The percentage of isolates resistant to various antimicrobials was not related to concurrent use of antimicrobials in the feed. CONCLUSIONS AND CLINICAL RELEVANCE: With the exception of tetracycline and sulfamethoxazole, resistance of Salmonella isolates to any of the antimicrobials was uncommon. Most isolates were susceptible to all antimicrobials tested. Antimicrobial resistance was not related to the presence of antimicrobials in the ration being fed at the time of sample collection.

Animals↗

A double antibody sandwich enzyme-linked immunosorbent assay for the detection of Salmonella using biotinylated monoclonal antibodies.

A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed using monoclonal antibodies (MAbs) as a rapid, economical alternative to culture isolation procedures for detection of Salmonella. Four MAbs previously shown to react with Salmonella strains representing 18 different serogroups were evaluated as capture antibodies and, after biotinylation, as detection antibodies. One MAb (M183) was selected for use in the ELISA to capture and detect Salmonella antigens. The detection limit of the ELISA was evaluated using Salmonella enterica subspecies enterica serovar Typhimurium and various selective and nonselective Salmonella enrichment media. The highest detection limit (ca. 10(4) CFU/ml) was achieved using an enrichment broth containing brain heart infusion, yeast extract, sodium hydrogen selenite, and sodium cholate (BYSC) after preenrichment in buffered peptone water. The ELISA detected all Salmonella serovars tested, which included representative serovars of serogroups B, C, D, and E and gave negative results for all non-Salmonella species tested. Samples (106) from various sources, including fecal samples from humans and pigeons, chicken carcass rinses, chicken parts, feed, and the environment, were used to evaluate the performance of the ELISA. The ELISA had a specificity and sensitivity of 100 and 91%, respectively, and a kappa value of 0.93 relative to the culture methods. Such an ELISA has the potential to be used in the implementation of the pathogen reduction and hazard analysis critical control point systems as well as in clinical laboratories.

Antibodies, Monoclonal↗

Detection of Salmonella dublin mammary gland infection in carrier cows, using an enzyme-linked immunosorbent assay for antibody in milk or serum.

An ELISA has been developed for measurement of milk and serum IgG concentrations directed against Salmonella dublin. Four groups of cows were studied: group A--7 experimentally challenge-exposed cows (infected, recovered group); group B--6 normal uninfected randomly selected control cows; group C--7 naturally occurring S dublin carrier cows; and group D--6 normal uninfected S dublin negative cows from the same herd as group C. Group-A cows were inoculated orally, or inoculated orally and then IV, but none became a S dublin carrier. As expected, all 7 group-A cows responded with a marked increase in ELISA titer after oral exposure to virulent S dublin, starting with a mean serum titer of 17.7% and reaching a peak mean serum titer of 79.3% approximately 76 days after initial exposure. As determined by necropsy and organ culturing of the remaining cows, none of the group-A cows became carriers. The mean serum ELISA titer for group-B uninfected control cows was 14.1% (SD +/- 12.8%). The mean milk ELISA titer was -1.0% (SD +/- 5.5%). Colostrum and then milk gave false-positive results for up to 2 weeks after onset of lactation. Group-B cows were culture negative for S dublin in feces and milk during lactation, and when tissues were cultured after euthanasia. Milk and serum samples for ELISA, and milk and fecal samples for culturing were taken from all group-A and -B cows twice a week for 6 months. Statistical correlation (P less than 0.05) was found between serum and milk ELISA titers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗