[Comparative tests to demonstrate thermolabile enterotoxins in strains of E. coli isolated from samples of fecal matter].
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Diarrhea↗
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Influence of delay in stool sampling on fecal occult blood test sensitivity.
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Feces↗
Comparison of sampling methods for microbiological testing of beef animal rectal/colonal feces, hides, and carcasses.
This study compared sampling methods for detecting Escherichia coli O157:H7 and Salmonella in beef cattle feces and on hides and carcasses and for enumerating E. coli biotype I counts (ECC) on carcasses. Fecal samples were collected by rectal/colonal palpation and colonal sponge swabbing. Hides were sampled by sponge swabbing three sites, hair clipping, excision, rinsing, and gauze swabbing, whereas carcasses were sampled by three-site thoracic and pattern-mark sponge swabbing and tissue excision. Overall, irrespective of sampling method, 36.7, 13.3, and 0.0% of lots contained at least one E. coli O157:H7-positive hide, fecal, and carcass sample, respectively, while the corresponding prevalence of Salmonella was 70.0, 16.7, and 6.7%, respectively. For hide sampling, excision and gauze swabbing yielded the fewest (13.3%) E. coli O157:H7-positive samples, while hair clipping and sponge swabbing yielded the most (23.3%). None of the carcass-sampling methods detected E. coli O157:H7 or differed (P > 0.05) in their ability to enumerate ECC. Colonal swabbing was the most effective (10.0%) method for detecting E. coli O157:H7 in feces. No differences (P > 0.05) in Salmonella prevalence were observed between carcass-sampling methods, although three-site sponge swabbing and tissue excision detected the most (3.3%). Hide rinsing was the most effective (P < 0.05) Salmonella detection method (63.3%), but dangers associated with its application may preclude its use by industry; there were no differences (P > 0.05) among other hide-sampling methods. No differences (P > 0.05) in Salmonella detection were observed between fecal-sampling methods. Overall, three-site sponge swabbing was the most feasible and effective sampling method for the detection of E. coli O157:H7 and Salmonella on hides and carcasses.
Animals↗
Comparison of pharyngeal, fecal, and cloacal samples for the isolation of Chlamydia psittaci from experimentally infected cockatiels and turkeys.
Direct comparisons were made of chlamydial isolation rates from pharyngeal swabs, fecal samples, and cloacal swabs from cockatiels and pharyngeal and cloacal swabs from turkeys experimentally infected with Chlamydia psittaci. During pathogenesis studies, 133 paired specimens were collected from cockatiels and 118 paired specimens were collected from turkeys. Of the 51 cockatiel chlamydial infections detected, 80.4% were positive by the pharyngeal swab sample, 45.1% were positive by the fecal swab sample, and 37.3% were positive by the cloacal swab sample. Of the 87 turkey infections detected, 93.1% were positive by the pharyngeal swab sample and 77.0% were positive by the cloacal swab. The pharyngeal swabs were the most reliable sample for isolation of chlamydia from live birds. However, no single sampling site yielded positive results from all infected birds. Specimens from multiple sites are recommended because a number of infected birds were identified by isolation from only 1 sample.
Animals↗
Fecal calprotectin levels in healthy children studied with an improved assay.
BACKGROUND: Calprotectin is a protein abundant in neutrophils. Fecal calprotectin can be used as a marker of gastrointestinal inflammation, and an improved assay has recently been developed. The aim of this study was to establish reference values for fecal calprotectin in healthy children aged between 4 and 17 years. METHODS: Fecal samples were obtained from 117 healthy children classified into four age groups: 4 to 6 years, 7 to 10 years, 11 to 14 years, and 15 to 17 years. A health questionnaire was used to ensure that these children fulfilled the inclusion criterion and did not have intercurrent disease, nasal or menstrual bleeding, or nonsteroidal anti-inflammatory drug medication before the sampling period. Calprotectin was analyzed using a quantitative enzyme-linked immunosorbent assay (Calprest, Eurospital SpA, Trieste, Italy). Children with fecal calprotectin values >50 microg/g were asked to deliver an additional sample. RESULTS: The overall median fecal calprotectin concentration was 13.6 microg/g (95% confidence interval, 9.9-19.5 microg/g) in the 117 children. In the different age groups, 4 to 6 years, 7 to 10 years, 11 to 14 years, and 15 to 17 years, the median calprotectin concentrations were 28.2, 13.5, 9.9, and 14.6 microg/g, respectively. Of these children, 104 (89%) had a concentration <50 microg/g. The remaining 13 children with a calprotectin concentration >50 microg/g delivered one additional fecal sample. All showed a lower concentration in the second sample except for one teenager who later proved to have proctitis. CONCLUSIONS: The suggested cutoff level for adults (<50 microg/g) can be used for children aged from 4 to 17 years regardless of sex. A fecal calprotectin concentration >50 microg/g warrants follow-up.
Adolescent↗
Persistence of colonization of human colonic mucosa by a probiotic strain, Lactobacillus rhamnosus GG, after oral consumption.
Lactobacillus rhamnosus GG is one of the most thoroughly studied probiotic strains. Its advantages in the treatment of gastrointestinal disorders are well documented. The aim of the present study was to demonstrate with colonic biopsies the attachment of strain GG to human intestinal mucosae and the persistence of the attachment after discontinuation of GG administration. A whey drink fermented with strain GG was fed to human volunteers for 12 days. Fecal samples were collected before, during, and after consumption. L. rhamnosus GG-like colonies were detected in both fecal and colonic biopsy samples. Strain GG was identified by its characteristic colony morphology, a lactose fermentation test, and PCR. This study showed that strain GG was able to attach in vivo to colonic mucosae and, although the attachment was temporary, to remain for more than a week after discontinuation of GG administration. The results demonstrate that the study of fecal samples alone is not sufficient in evaluating colonization by a probiotic strain.
Administration, Oral↗
Variation in Eimeria oocyst count and species composition in weanling beef heifers.
Rectal fecal samples were collected daily on 10 consecutive days in November 2004 from 11 weaned beef heifers to assess daily variation in fecal oocyst count and species composition. Subsequent samples were collected from the same animals on 15 April 2005 and 9 June 2005. Oocyst numbers were determined by the modified McMaster's test, and species were identified by examination of oocysts recovered with the Wisconsin sugar flotation technique. Soil samples were collected from the heifer pasture on 8 June 2005, and oocysts were quantified and identified to species. Mean fecal oocyst counts varied little at all sampling dates ranging from 134-377 oocysts/g. Ten Eimeria spp. were identified in fecal samples collected in November and April and 11 in June. Eimeria bovis was the most common species identified at all samplings. Mean species composition showed little variation during the 10-day sampling period in November, remained similar in April, and varied slightly in June. Twelve Eimeria spp. were identified in soil samples in proportions similar to those seen in fecal samples. The results indicate that clinically normal weanling beef heifers are likely to be infected with a diverse, but relatively stable, community of Eimeria spp.
Animals↗
Experiments in DNA extraction and PCR amplification from bighorn sheep feces: the importance of DNA extraction method.
Reliability of genotyping is an issue for studies using non-invasive sources of DNA. We emphasize the importance of refining DNA extraction methods to maximize reliability and efficiency of genotyping for such DNA sources. We present a simple and general method to quantitatively compare genotyping reliability of various DNA extraction techniques and sample materials used. For bighorn sheep (Ovis canadensis) fecal samples we compare different fecal pellet materials, different amounts of fecal pellet material, and the effects of eliminating two DNA extraction steps for four microsatellite loci and four samples heterozygous at each locus. We evaluated 192 PCR outcomes for each treatment using indices of PCR success and peak height (signal strength) developed from analysis output of sequencer chromatograms. Outermost pellet material produced PCR results almost equivalent to DNA extracted from blood. Where any inner pellet material was used for DNA extraction, PCR results were poorer and inconsistent among samples. PCR success was not sensitive to amount of pellet material used until it was decreased to 15 mg from 60 mg. Our PCR index provides considerably more information relative to potential genotyping errors than simply comparing genotypes derived from paired fecal and blood or tissue samples. Our DNA extraction method probably has wide applicability to herbivores that produce pelleted feces where samples dry rapidly after deposition.
Animals↗
Occurrence of Salmonella serotype Typhimurium DT104 on a commercial swine farm before, during, and after depopulation and repopulation.
OBJECTIVE: To determine whether depopulation-repopulation could be used to eradicate Salmonella serotype Typhimurium DT104 from a commercial swine farm in the midwestern United States. DESIGN: Observational study SAMPLE POPULATION: A commercial swine farm undergoing depopulation-repopulation to eliminate porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae. PROCEDURE: Pooled fecal samples, tissue samples, and serum samples were collected from pigs on the farm before and after depopulation-repopulation. When there were no pigs on the farm, environmental swab specimens were collected for bacterial culture. Serum was analyzed for anti-Salmonella antibodies with an indirect ELISA. Salmonella isolates obtained by bacterial culture of fecal, tissue, and environmental samples were characterized by means of serotyping, phage typing, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing. RESULTS: 167 Salmonella isolates representing 9 serotypes were recovered from the farm. Results of PFGE and antimicrobial susceptibility testing suggested that S. Typhimurium DT104 strain was not eradicated from the farm. However, seroprevalence of anti-Salmonella antibodies and the percentage of pooled fecal samples positive for Salmonella spp were significantly decreased following repopulation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that depopulation-repopulation in conjunction with stringent cleaning and disinfection, attention to biosecurity procedures, control of other diseases, and changes in feed management may reduce the occurrence of, but likely will not eliminate, Salmonella spp in commercial swine herds.
Animal Husbandry↗
Molecular typing of VapA-positive Rhodococcus equi isolates from Jeju native horses, Korea.
We recently demonstrated the presence of virulence-associated protein antigen (VapA)-positive Rhodococcus equi in Jeju Island, Korea. These bacteria contained one of two distinct plasmid types, a 90-kb type II plasmid, which has been found in isolates from the native Kiso horses of Japan, and a new variant, a 90-kb type V plasmid. However, the genotypic characters of the VapA-positive R. equi from Jeju native horses and their environments are poorly understood. Ninety-eight isolates from soil samples and 89 isolates from fecal samples were collected from five farms that breed or have bred Jeju native horses, and were tested for the presence of VapA by immunoblotting and PCR. Of the 98 soil isolates and 89 fecal isolates, seven and 13 were VapA-positive R. equi, respectively. In 2003, two Jeju foals died suddenly and were brought to the Faculty of Veterinary Medicine, Cheju National University, for postmortem examination. Pure cultures of R. equi were isolated from the lung lesions of both foals. Of the 16 clinical isolates, 14 were VapA-positive R. equi. Of the 34 VapA-positive clinical and environmental isolates, 16 contained the 90-kb type II plasmid and 18 contained a 90-kb type V plasmid. Pulsed-field gel electrophoresis (PFGE) patterns of the VapA-positive isolates from Jeju horses and Kiso horses, containing the 90-kb type II plasmid, were compared and formed two distinct groups. Furthermore, 18 virulent isolates containing the 90-kb type V plasmid formed two distinct PFGE groups (of 16 and two isolates). These results demonstrate that two virulence plasmid types are widespread in R. equi in Jeju native horses. However, there is little diversity in the PFGE patterns of virulent isolates, suggesting the clonal spread of virulent R. equi. The PFGE pattern of the virulent R. equi isolates from Jeju native horses in Korea is not identical to those of Kiso native horses in Japan.
Actinomycetales Infections↗
Comparison of rectal swabs with fecal cultures for detection of Salmonella typhimurium in adult volunteers.
Rectal swabs are generally considered less sensitive than fecal culture, but there are no data directly comparing human rectal swabs with fecal samples for detection of Salmonella. A phase I clinical study of a live oral attenuated Salmonella typhimurium vaccine strain in volunteers receiving a large known inoculum provided the opportunity to compare concurrent rectal swab and fecal cultures. Of 155 paired samples from 9 volunteers, 65 (42%) were culture positive: 35 (54%) by both methods, 20 (31%) by fecal culture only, and 10 (15%) by swab only. When compared with fecal culture, rectal swabs were 64% sensitive and 90% specific. Rectal swabs are of moderate diagnostic utility for detection of Salmonella and may be useful when collection of fecal samples is impractical.
Adult↗
Occurrence of microbial indicators and Clostridium perfringens in wastewater, water column samples, sediments, drinking water, and Weddell seal feces collected at McMurdo Station, Antarctica.
McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered.
Animals↗
Relationship between ovarian cycle phase and sexual behavior in female Japanese macaques (Macaca fuscata).
We conducted behavioral observations simultaneously with fecal sample collection on eight nonlactating females 2-3 times per week, October 1997-March 1998, to examine the relationship between ovarian hormones and the sexual behavior of female Japanese macaques (Macaca fuscata) during the mating season. We analyzed samples by enzyme immunoassay for fecal hormone levels. Hormone profiles of estrone-glucuronide (E1) and pregnanediol-glucuronide (PdG) were used to separate ovarian cycles into three phases (follicular, periovulatory, and luteal). Hormonal profiles indicate average cycle lengths of 27.6 +/- 4.2 days (+/- SD; n = 26). Average lengths of the luteal and follicular phases were 12.3 +/- 3.8 days (+/- SD) and 8.3 +/- 3.4 days (+/- SD), respectively. We observed female Japanese macaques engaging in sexual activity throughout the ovarian cycle, with the highest rates occurring during the follicular and periovulatory phases as compared to the luteal phase. The attractivity of female Japanese macaques increased significantly during the follicular and periovulatory phases of the ovarian cycle, when E1 levels are peaking and PdG levels drop to baseline. In addition, females displayed a significant increase in proceptive behavior during the follicular and periovulatory phases. Grooming bouts, as well as proximity between female and male macaques, also increased significantly during the follicular and periovulatory phases. We conclude that fluctuating levels of ovarian hormones in different phases of the cycle are significantly associated with variable rates of copulatory and pericopulatory behaviors in these Japanese macaque females.
Animals↗
Factors associated with ileal mucosal morphology and inflammation in patients with ileal pouch-anal anastomosis for ulcerative colitis.
PURPOSE: Pouchitis has been associated with abnormal bacterial flora responding to antibiotics. Dietary factors may play a role in modifying the qualitative and quantitative components of the microflora. We evaluated interactions between nutritional factors, fecal and mucosal bacterial flora, and mucosal morphology in patients with a history of pouchitis compared with patients with optimal outcome at least five years after ileal pouch-anal anastomosis for ulcerative colitis. METHODS: Thirty-two patients were enrolled in the study: 11 (7 males; mean age, 49.8 years) with optimal outcome and 21 (11 males; mean age, 47.3 years) with pouchitis history. A seven-day food diary was recorded, endoscopy performed, and biopsies taken from the pouch for histology, mucin staining, and bacterial culture. Fresh fecal samples were quantitatively cultured, and fecal bile acids analyzed by gas-liquid chromatography. RESULTS: No differences existed in mean nutrient intake, composition of fecal bile acids, or microbial tissue biopsy cultures between the groups with and without pouchitis. Those with optimal outcome tended to have more benign disease course of ulcerative colitis than patients with pouchitis. In those patients, fecal concentrations (log10 colony-forming unit/g) of anaerobes and aerobes were significantly higher (P = 0.007). Degree of villous atrophy and colonic metaplasia were both associated with fecal anaerobic flora. Low intake of lactose was associated with sulfomucin predominance. A negative correlation existed between fecal aerobes and dietary lactose consumption. CONCLUSIONS: A higher total load of fecal anaerobic bacterial flora is strongly associated with degree of colonic metaplasia, villous atrophy, and inflammation activity after surgery for ulcerative colitis. An association existed between dietary lactose, fecal bacteria, and pouch morphology. Lactose may have prebiotic properties.
Adult↗
Prevalence of intestinal parasites in dogs under veterinary care in Maracaibo, Venezuela.
The prevalence of intestinal parasites in dogs presented to the Veterinary Policlinic of the University of Zulia (PVU) was measured between January and December 2001. A total of 614 fecal samples were evaluated by the fecal flotation method. One or more species of parasites was identified in 218 (35.5%) dogs. The parasites most frequently detected were: Ancylostoma spp. (24.5%), Toxocara canis (11.4%) and Isospora spp. (8.1%). Single parasitic infections were present in 149 (24.3%) dogs. The age distribution of intestinal parasites in dogs less than 1 year old had a higher overall prevalence than those dogs over 12 months of age. There was no significant difference in the prevalence between male (38.9%) and female (31.7%) dogs. There was a significantly (P < 0.05) greater prevalence of parasites in mixed-breed dogs (40.3%) as compared with pure-breed dogs (30.8%). A significant difference (P < 0.05) was detected between the general prevalence of January and December compared to August.
Age Factors↗
Assessing reproductive status of right whales (Eubalaena glacialis) using fecal hormone metabolites.
Long-term studies of the endangered North Atlantic right whale, Eubalaena glacialis, have revealed declining reproductive parameters over the past two decades, threatening recovery of this small population if current trends continue. Little is known about right whale reproductive physiology, and investigating this reproductive decline has been limited by a lack of non-lethal methods for assessing reproductive status (e.g., sexual maturation, ovarian activity, pregnancy, lactation, and reproductive senescence) in free-swimming whales. This paper describes validation of existing radioimmunoassay techniques to study reproduction in right whales by measuring estrogens, progestins, androgens, and their related metabolites in fecal samples. Over the past decade fecal steroid hormone assays have been used to assess reproductive status and function in a wide range of terrestrial wildlife species, but this is the first application of this methodology in wild cetaceans. Analysis of fecal hormone metabolite levels in combination with life history data from photographically identified whales shows that this non-invasive method can be used to determine gender, detect pregnancy and lactation, and to assess age at sexual maturity in right whales and potentially other endangered whale populations.
Androgens↗
Whey proteins protect more than red meat against azoxymethane induced ACF in Wistar rats.
Protein type and density have been shown to influence colon cancer risk using a carcinogen-induced rat model. It is suggested that red meat may promote colon cancer risk more than whey proteins. The aim of this study was to evaluate the influence of red meat, whey protein and their density in the diet on the number of aberrant crypt foci (ACF), preneoplastic markers in Wistar rats. The sources of protein, red meat as barbecued kangaroo muscle meat, and whey protein concentrate were fed to rats to provide 8, 16 and 32% protein by weight in a modified AIN-93 diet with low fiber, low calcium and high polyunsaturated fat. Adult Wistar rats (13 weeks of age) were fed these diets for 4 weeks and then two s.c. injections of azoxymethane, 15 mg/kg BW, were administered 1 week apart. Diets were fed for a further 8 weeks, rats were then killed, their colons fixed in formalin saline and stained with methylene blue to quantify ACF number. Fecal samples were collected and the fecal water was isolated for quantification of heme and thiobarbituric acid reactive substances. Increasing red meat density correlated positively, while increasing dairy protein density correlated negatively with rate of weight gain (p<0.05). Dietary intake was not significantly affected by protein type or density. The 32% whey protein group had significantly less ACF in the proximal colon in comparison to the 16 and 32% red meat groups (p<0.05). This reduction in ACF number in the whey protein group may be caused by hormones associated with the reduction in weight gain, and/or by components of whey protein concentrate such as cysteine, lactose and conjugated linoleic acid which have been shown to have anti-cancer effects. Using ACF number as an index, whey protein appeared to be more protective than red meat.
Animals↗
Use of the regression analysis technique to determine the true phosphorus digestibility and the endogenous phosphorus output associated with corn in growing pigs.
The objectives of this study were to determine true phosphorus (P) digestibility and the endogenous P output associated with corn for growing pigs using the regression analysis technique. Four barrows, average initial body weight of 25 kg, were fitted with a T-cannula and fed four diets according to a 4 x 4 Latin square design. Four cornstarch-based diets, containing four levels of P at 0.7, 1.5, 2.2 and 2.8 g/kg dry matter intake (DMI), were formulated from corn. Each experimental period comprised 8 d with 4-d adaptation and 4-d collection of ileal digesta and fecal samples. The apparent ileal and fecal P digestibility values in corn were affected (P < 0.05) by P contents in the assay diets. The apparent ileal and fecal P digestibility values increased from -51.0 to 33.3% and from -41.4 to 39.1%, respectively, as P content increased from 0.7 to 2.8 g/kg DMI. Linear relationships (P < 0.05), expressed as g/kg DMI, between the apparent ileal and fecal digestible P and the total intake of dietary P, suggested that true P digestibility and the endogenous P outputs associated with corn can be determined by the regression analysis technique. There were no differences (P > 0.05) in true P digestibility values (54.0 +/- 6.5 vs. 59.8 +/- 8.5%) and the endogenous P outputs (0.693 +/- 0.128 vs. 0.670 +/- 0.160 g/kg DMI) between the ileal and the fecal determinations. The endogenous fecal P output represented 12.3% and 25.8% of the daily total and available P requirements in growing pigs recommended by the National Research Council in 1998. The present literature data of apparent digestibility and availability underestimate the true digestive utilization of P in corn for growing pigs by approximately 35%. Current diet formulation on the bases of total, apparent P digestibility and availability values in corn inevitably leads to P overfeeding and excessive P excretion in pigs.
Animals↗