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Blood samples, fecal samples, and cloacal swabs were collected from 42 bog turtles (Clemmys muhlenbergii). including 14 wild males, 22 wild females, three captive males, and three captive females, in Virginia and North Carolina, USA. Samples were analyzed for hematologic and plasma chemistry values, Mycoplasma sp. antibodies, intestinal parasites, and normal cloacal flora.
Twenty-one genetically lean, obese or contemporary slaughter weight castrated male pigs (6 mo old; seven of each genotype) were assigned to individual tether stalls and fed either a control diet (low fiber) or a diet containing 80% alfalfa meal (high fiber) at 1.50% of initial body weight for 71 d (1.75% for d 1 to 4). Apparent dry matter digestibility of the diets was estimated by determining acid insoluble ash in fecal samples. Fecal cellulolytic bacteria and total viable bacteria were enumerated at d 0, 14, 35, 49 and 70. Fecal inocula were used to determine 48-h in vitro digestibility of alfalfa meal fractions on the same days. Digesta rate of passage was determined by feeding a pulse dose of chromium-mordanted alfalfa fiber to the pigs fed the high-fiber diet. In vivo digestibility of both diets was less for the obese pigs than for the lean or contemporary genotypes. In vitro digestibility of alfalfa fiber fractions was not different between the genotypes fed either diet. When the high-fiber diet was fed, in vitro digestibility increased for all genotypes from d 0 to d 14, but not thereafter. The numbers of cellulolytic bacteria for all three genotypes were greater when pigs were fed the high-fiber diet (23.0 X 10(8), 51.6 X 10(8), 37.2 X 10(8) per gram fecal dry weight; obese, lean and contemporary, respectively) compared to the low-fiber diet (3.0 X 10(8), 3.2 X 10(8), 3.4 X 10(8), respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Fecal glucocorticoid metabolite analysis is a useful tool for monitoring adrenocortical activity in captive and free-ranging wildlife. Glucocorticoid metabolites may not be evenly distributed within fecal samples and this variability could affect the interpretation of glucocorticoid metabolite measurements. Furthermore, the precision (i.e., repeatability of measurements) of fecal glucocorticoid measurements from well-mixed samples is unknown. We collected fresh white-tailed deer (Odocoileus virginianus) feces at various times pre- and post-adrenocorticotropin injection to provide samples with low (<75 ng/g), medium (75-90 ng/g), and high (>90 ng/g) glucocorticoid concentrations in case variability differs in samples of dissimilar hormone metabolite concentrations. We compared two sampling methods (selection of three pellet groups [one from each end of the fecal mass and one from the center] versus sampling three small portions of the thoroughly mixed fecal mass) to estimate within-sample variation of glucocorticoid metabolites. Glucocorticoid metabolite measures from pellet groups were higher than fecal glucocorticoid measures from mixed samples in the low (F=3.10; df = 1,56; P=0.08) and medium concentration (F=7.28; df = 1,50; P=0.01) groups. Fecal glucocorticoid metabolite estimates from mixed samples were less variable than glucocorticoid metabolite measures using pellet groups from the same fecal mass, although these differences were not statistically significant (low group: F=0.59; df = 1,38; P=0.45; medium group: F=0.13; df = 1,34; P=0.72; high group: F=2.30; df = 1,28; P=0.14). The mean coefficient of variation was <10% across all treatment groups and sampling methods. However, a power analysis indicated the mixed sub-sample technique requires fewer samples to detect statistically significant differences than pellet groups. Our results suggest that glucocorticoid metabolites may not be evenly distributed in white-tailed deer feces. Consequently, using only a few pellets from a fecal mass may bias assay interpretation. We suggest researchers homogenize the entire fecal mass before removing a sub-sample of fecal material for analysis. Also, other sources of variation must be considered when interpreting results of fecal glucocorticoid studies.
Bacteroides fragilis is an obligatory anaerobic, gram-negative bacterium found among the normal intestinal flora of humans. Enterotoxigenic strains of B. fragilis (ETBF) have been associated with diarrheal diseases in humans and animals. The enterotoxin of ETBF induces fluid changes in ligated intestinal segments and cytotoxic response in HT29/C1 cells. By using a pair of monoclonal antibodies (MAbs; MAb C3 and MAb 4H8) specific for the lipopolysaccharide of B. fragilis, an assay based on immunomagnetic separation (IMS) in combination with PCR (IMS-PCR) was developed. After DNA extraction, a 294-bp fragment was amplified. The specificity of the IMS-PCR assay was evaluated by adding previously isolated and confirmed ETBF strains to normal fecal samples. All fecal samples to which ETBF strains were added were positive, showing a 100% specificity. The spiked fecal samples were also used for evaluation of the sensitivity of the assay. The detection limit was found to be approximately 50 CFU/g of feces. By this method 10 clinical fecal samples (5 from patients with diarrhea and 5 from healthy controls) were examined. The results of PCR were in accordance with the results of the HT29/C1 cell assay for all samples. The minimum time to retrieval of the final result by the IMS-PCR method is 36 h. The proposed IMS-PCR assay is rapid and sensitive for the direct detection of ETBF in stool samples.
OBJECTIVE: To compare antimicrobial susceptibility patterns of Escherichia coli isolates cultured from fecal samples from cows and calves on dairy farms that used organic (ie, no or severely limited antimicrobial use) versus conventional production methods. DESIGN: Cross-sectional study. SAMPLE POPULATION: Fecal samples from 10 cows and 10 calves on each of 30 organic dairy farms and 30 neighboring conventional dairy farms in Wisconsin. PROCEDURE: E. coli isolates obtained from the fecal samples were tested for susceptibility to 17 antimicrobials by means of a microbroth dilution test. Prevalence of antimicrobial resistance was compared between organic and conventional dairy farms. RESULTS: E. coli was isolated from 1,121 (94%) fecal samples. Farm type (organic vs conventional) and animal age (cow vs calf) were significantly associated with odds that E. coli isolates would be resistant to various antimicrobials. After controlling for age, logistic regression analyses indicated that isolates from conventional dairy farms had significantly higher rates of resistance to ampicillin, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, and sulfamethoxazole than did isolates from organic dairy farms. However, no significant differences were detected for the 10 other antimicrobials that were tested. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that compared with isolates from conventional dairy farms, E. coli isolates from organic dairy herds have significantly lower prevalences of resistance to 7 antimicrobials; however, prevalence of resistance was not significantly different for 10 other antimicrobials. Resistance was more common for isolates from calves than for isolates from adult dairy cows.
Bovine coronavirus (BCoV) is an etiological agent associated with winter dysentery (WD), prevalent in adult cattle during the winter. Although we previously detected, isolated, and characterized BCoV strains from adult cattle with WD (WD-BCoV strains) during the winter in South Korea, the precise epidemiology, as well as the causative agent of diarrhea in adult cattle in the warmer seasons, has not been examined. We examined 184 diarrheic fecal specimens collected from 75 herds of adult cattle from seven provinces during the spring (warm), autumn (warm), and summer (hot) seasons. Bovine coronavirus-positive reactions were detected for 107 (58.2%) diarrheic fecal samples (in 47/75 herds). Of these 107 positive samples, 90 fecal samples from 33 herds tested positive for BCoV alone and 17 fecal samples from 14 herds also tested positive for other pathogens. Biological comparisons between the 9 BCoV strains isolated in this study and the 10 previously isolated WD-BCoV strains revealed that there was no receptor-destroying enzyme (RDE) activity against mouse erythrocytes in the 9 BCoV strains but the 10 WD-BCoV strains had high RDE activity. Phylogenetic analysis of the spike (S) and hemagglutinin/esterase (HE) proteins revealed that all the Korean BCoVs clustered together regardless of season and were distinct from the other known BCoVs, suggesting a distinct evolutionary pathway for the Korean BCoVs. These and previous results revealed a high prevalence and widespread geographical distribution of BCoV, suggesting that this virus is endemic in adult cattle with diarrhea in all seasons in South Korea.
This study was designed to estimate the contribution of municipal sewage sludge exposure and smoking to the daily Cd intake in Ohio farm residents, based on the Cd concentrations in 24-hr fecal samples. The fecal samples were analyzed for Cd and the daily Cd intakes were calculated by correcting for absorption (4.6%) in the gastrointestinal tract. Fecal samples from cattle grazing on sludge-treated pastures were also similarly treated except that published 24-hr fecal weights and a 2% absorption correction were used. Fecal weights and the daily Cd intakes, in humans, calculated from these data were significantly lower in females than in males with a female/male ratio of 0.77/1. Daily Cd intakes calculated from these data ranged from 5.37 to 13.31 micrograms/day for females and 8.87 to 18.52 micrograms/day for males. No significant increase in daily Cd intake resulted from exposure of humans to sludge on the farmlands. Although daily Cd intake for smokers was 1 microgram/day higher than for non-smokers, the difference was not statistically significant. Cattle grazing on sludge-treated pastures consumed significantly more (up to 3 times) Cd than cattle on control pastures. It was concluded that application of sewage sludge on farmlands at rates of 2-10 dry metric tons/ha did not significantly contribute to the daily Cd intake in humans; cattle on such farms, however, significantly increased their Cd consumption.
OBJECTIVE: To compare 3 alternative culture techniques for the detection of Salmonella organisms in swine feces with a modification of the International Standard Organization (ISO) 6579 standard protocol. SAMPLE POPULATION: Fecal samples from swine herds suspected of having Salmonella infections. PROCEDURE: 4 experiments were performed to evaluate the following: 1) diagnostic sensitivity of the selective preenrichment and rapid isolation novel technology (SPRINT) protocol, compared with that of the modified ISO protocol; 2) detection limit of the SPRINT protocol for Salmonella organisms; 3) use of tetrathionate-novobiocin (TTN) broth, compared with selenite cysteine (SC) broth for selective enrichment; and 4) use of universal preenrichment (UPE) broth, compared with buffered peptone water (BPW) for preenrichment of samples prior to the use of modified semisolid Rappaport-Vassiliadis (MSRV) plates. RESULTS: Comparing the Salmonella culture results of 183 swine fecal samples, the diagnostic sensitivity of the SPRINT protocol (0.86) was not significantly different than the diagnostic sensitivity of the modified ISO protocol (0.80), although it was 24 hours faster. The SPRINT protocol could detect 5 of the 6 investigated Salmonella serotypes at inoculation concentrations of < 10 colony-forming units (CFU)/25 g of uncontaminated feces. The TTN broth performed significantly better than the SC broth for selective enrichment of Salmonella organisms. There was no significant difference in results of preenrichment of samples between the use of UPE broth or BPW. CONCLUSIONS AND CLINICAL RELEVANCE: The SPRINT protocol may provide a faster alternative for isolation of Salmonella organisms from swine fecal samples. Furthermore, the use of TTN broth instead of SC broth may increase the sensitivity of the modified ISO 6579 protocol.
OBJECTIVE: To evaluate 2 rapid, patient-side assays for detection of Cryptosporidium parvum in feces from neonatal calves with diarrhea. DESIGN: Diagnostic test evaluation Sample Population-Fecal samples from 96 neonatal (1 to 30 days old) calves with diarrhea. PROCEDURE: Results of the rapid assays were compared with results of microscopic examination of fecal smears that had been stained with diamant fuchsin stain. RESULTS: One of the rapid assays correctly identified 56 of 62 (90%) fecal samples positive for C. parvum oocysts and 33 of 34 (97%) fecal samples negative for oocysts. The other assay correctly identified 53 of 62 (85%) fecal samples positive for oocysts and 33 of 34 (97%) fecal samples negative for oocysts. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that these 2 rapid assays are accurate when used to detect C. parvum in fecal samples from neonatal calves with diarrhea.
OBJECTIVE: To isolate and speciate Cryptosporidium DNA from fecal samples obtained from dairy cattle in New York State and identify factors associated with whether cattle were shedding Cryptosporidium parvum versus Cryptosporidium bovis. DESIGN: Cross-sectional study. SAMPLE POPULATION: 115 fecal samples positive for DNA coding for the Cryptosporidium 18S rRNA gene from dairy cattle in New York State. PROCEDURES: A PCR assay was used to amplify DNA from fecal samples; amplification products were submitted for bidirectional DNA sequencing. Logistic regression was used to test for associations between various host factors and Cryptosporidium spp. RESULTS: 70 of the 115 (61%) fecal samples were found to have C parvum DNA, 42 (37%) were determined to have C bovis DNA, and 3 (3%) were found to have C parvum deer-type DNA. The presence of diarrhea at the time of fecal sample collection, oocyst count, and breed were associated with whether cattle were infected with C parvum or C bovis, with animals more likely to be infected with C parvum if they had diarrhea, had a high oocyst count, or were Holsteins. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that C parvum and C bovis can be isolated from dairy cattle in New York State and that various factors affect whether cattle infected with Cryptosporidium spp are infected with C parvum or C bovis. Findings also lend credence to the theory that C bovis may be more host adapted and thus less pathogenic to dairy cattle than C parvum.
A competitive enzyme-linked immunosorbent assay (ELISA) for detection of canine parvovirus (CPV) antigen in fecal samples was developed. Fecal samples were tested by ELISA and a direct hemagglutination assay, and the results compared. The tests gave the same results in 83% and 88% of the fecal samples, depending on whether the samples were treated with chloroform. The discrepancies were due to the fact that each test detected virus in certain fecal samples that was not detected by the other. The use of a monoclonal anti-CPV conjugate resulted in a highly specific test. The performance of the ELISA as a competitive assay also increased the reproducibility and sensitivity over that which could be obtained from a classic sandwich-type procedure.
OBJECTIVE: To determine sources of Salmonella organisms in a veterinary teaching hospital, compare bacterial culture with polymerase chain reaction (PCR) testing for detection of Salmonella organisms in environmental samples, and evaluate the effects of various disinfectants on detection of Salmonella organisms on surface materials. DESIGN: Prospective study. SAMPLE POPULATION: Fecal samples from 638 hospitalized horses and 783 environmental samples. PROCEDURE: Standard bacterial culture techniques were used; the PCR test amplified a segment of the Salmonella DNA. Five disinfectants were mixed with Salmonella suspensions, and bacterial culture was performed. Swab samples were collected from 7 surface materials after inoculation of the surfaces with Salmonella Typhimurium, with or without addition of a disinfectant, and submitted for bacterial culture and PCR testing. RESULTS: Salmonella organisms were detected in fecal samples from 35 (5.5%) horses. For environmental samples, the proportion of positive bacterial culture results (1/783) was significantly less than the proportion of positive PCR test results (110/783), probably because of detection of nonviable DNA by the PCR test. Detection of Salmonella organisms varied with the surface material tested, the method of detection (bacterial culture vs PCR testing), and the presence and type of disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the present study suggested that Salmonella organisms can be isolated from feces of hospitalized horses and a variety of environmental surfaces in a large animal hospital. Although recovery of Salmonella organisms was affected by surface material and disinfectant, bleach was the most effective disinfectant on the largest number of surfaces tested.
Red meat consumption causes a dose-dependent increase in fecal apparent total N-nitroso compounds (ATNC). The genotoxic effects of these ATNCs were investigated using two different Comet assay protocols to determine the genotoxicity of fecal water samples. Fecal water samples were obtained from two studies of a total of 21 individuals fed diets containing different amounts of red meat, protein, heme, and iron. The first protocol incubated the samples with HT-29 cells for 5 min at 4 degrees C, whereas the second protocol used a longer exposure time of 30 min and a higher incubation temperature of 37 degrees C. DNA strand breaks were quantified by the tail moment (DNA in the comet tail multiplied by the comet tail length). The results of the two Comet assay protocols were significantly correlated (r = 0.35, P = 0.003), however, only the second protocol resulted in detectable levels of DNA damage. Inter-individual effects were variable and there was no effect on fecal water genotoxicity by diet (P > 0.20), mean transit time (P = 0.588), or weight (P = 0.705). However, there was a highly significant effect of age (P = 0.019). There was no significant correlation between concentrations of ATNCs in fecal homogenates and fecal water genotoxicity (r = 0.04, P = 0.74). ATNC levels were lower in fecal water samples (272 microg/kg) compared to that of fecal homogenate samples (895 microg/kg) (P < 0.0001). Failure to find dietary effects on fecal water genotoxicity may therefore be attributed to individual variability and low levels of ATNCs in fecal water samples.
A longitudinal study was performed in 67 larger Swiss cow-calf farms from September 1996 through November 1997. The objectives of the study were to estimate prevalence and risk factors for colonization with potentially zoonotic enteropathogenic bacteria in younger calves and in calves at weaning age. The study included data from 395 calves with three to four fecal samples each. Fecal samples were analyzed for Campylobacter spp., verotoxin producing E. coli (VTEC), Yersinia spp. and Salmonella sp. Possible environmental and individual factors associated with colonization of these agents were examined. The calves were housed indoor during the first 3 months of life (winter 1996/1997). The prevalences within this time period were: C. coli 3.4%, C. fetus 15.5%, C. hyointestinalis 9.6%, C. jejuni 38.5%, VTEC 44.3% and Yersinia spp. 2%. At the end of the grazing season the prevalences at weaning (8-10 months of age) were: C. coli 1.7%, C. fetus 4.0%, C. hyointestinalis 25.9%, C. jejuni 13.3%, VTEC 38.2% and Yersinia spp. 0%. No salmonellae were present at any time of the study. The prevalences of C. jejuni and VTEC increased significantly within the first 3 months of life, whereas C. hyointestinalis decreased. None of the environmental factors such as housing or feeding had any consistent influences on colonization by the bacteria studied. VTEC, Campylobacter spp. and Yersinia spp. should probably be considered as normal inhabitants of the bovine intestinal tract. However, as they represent a source of gastrointestinal infections in humans, management factors limiting intestinal colonization of these bacteria should be considered in cow-calf operations.
OBJECTIVE: To determine whether cats exposed at a residence were infected with Mycobacterium bovis, whether the tuberculin skin test can identify cats infected with M bovis, and whether an ELISA could identify tuberculosis-infected cats. ANIMALS: 20 domestic cats exposed to a cat with laboratory-confirmed disseminated M bovis infection. PROCEDURE: Cats were administered a tuberculin skin test and monitored for 72 hours. Blood and fecal samples were collected. Cats were then euthanatized, and postmortem examinations were performed. Tissues were examined grossly and histologically for signs of mycobacteriosis. Pooled tissue samples and fecal samples were submitted for mycobacterial culture. Blood samples were examined for evidence of tuberculosis by use of a comparative ELISA. RESULTS: 4 cats had positive responses for the ELISA, and 2 cats had suspicious responses. All tuberculin skin tests yielded negative results. No gross or histologic lesions of tuberculosis were detected in any tissues, and mycobacteria were not isolated from tissues or feces obtained from the 20 cats. CONCLUSIONS AND CLINICAL RELEVANCE: All cats that had positive or suspicious responses for the ELISA were offspring of the cat with tuberculosis. Evidence of tuberculosis was not seen in other cats at the residence, the owner, or the attending veterinarian. The most likely source of tuberculosis for the infected cat was through the consumption of M bovis-infected wildlife carcasses or offal. Because M bovis is endemic in wildlife in northeastern Michigan, there is a risk of exposure to tuberculosis in companion animals, their owners, and attending veterinarians.
Eighty-two coyote fecal samples were collected from eastern Colorado and examined for coccidia. Sporocysts of Sarcocystis were observed in 17 of the samples and oocysts of Isospora ohioensis were observed in two samples. Ten fecal samples contained eimerian coccidia thought to be pseudoparasites of the coyote acquired while ingesting heavily infected rabbits.
OBJECTIVE: To evaluate fecal concentrations of selected genera of colonic bacteria in healthy dogs, and to investigate effects of dietary fructooligosaccharides (FOS) on those bacterial populations. ANIMALS: 6 healthy adult Beagles. PROCEDURE: Dogs were randomly assigned to 2 groups of 3 and fed an unsupplemented diet for 370 days. After 88 days, fecal samples were collected. Another fecal sample was collected from each dog 282 days later. Group A then received a diet supplemented with FOS, and group B continued to receive the unsupplemented diet. Twenty-eight to 29 days later, fecal samples were collected. Diets were switched between groups, and fecal samples were collected 31 and 87 days later. Concentrations of Bifidobacterium spp, Lactobacillus spp, Clostridium spp, Bacteroides spp, and Escherichia coli in freshly collected feces were determined. Effects of diet and time on bacterial concentrations were compared between groups. RESULTS: Bifidobacterium spp and Lactobacillus spp were inconsistently isolated from feces of dogs fed either diet. Sequence of diet significantly affected number of Bacteroides spp subsequently isolated from feces, but diet had no effect on numbers of Clostridium spp or E coli. CONCLUSIONS AND CLINICAL RELEVANCE: Some genera of bacteria (eg, Bifidobacterium) believed to be common components of colonic microflora may be only sporadically isolated from feces of healthy dogs. This deviation from expected fecal flora may have implications for the effectiveness of supplementing diets with prebiotics.
Environmental conditions may influence fecal glucocorticoid metabolite measurements if feces cannot be collected immediately after deposition. To evaluate the influence of environmental conditions on fecal glucocorticoid metabolite concentrations, we exposed fresh fecal samples to 1 of 5 simulated conditions: (1) room temperature (22 degrees C), (2) high heat (38 degrees C), (3) alternating high heat and room temperature cycle, (4) alternating freezing (-20 degrees C) and room temperature cycle, and (5) simulated rainfall (0.85 cm every other day at 22 degrees C) for 7 days. We collected fresh white-tailed deer (Odocoileus virginianus) feces at various times pre- and post-adrenocorticotropin injection to provide samples with initially low (n=5), medium (n=5), and high (n=5) glucocorticoid concentrations. Feces were mixed thoroughly and then allocated into five 10-g samples. Also, a 5-g sub-sample was taken from each fecal mass prior to treatment and stored at -20 degrees C until assayed. We subsampled from all treatments once every 24-h for 7 days. Fecal samples were assayed using [125I]corticosterone radioimmunoassay kits. Fecal glucocorticoid metabolites in all three groups in the simulated rainfall treatment and the low group in the alternating freezing and room temperature treatment increased significantly over the 7-day period. We believe increased microbial metabolism of fecal glucocorticoids may partly explain these results. Other biochemical processes (e.g., cleavage of conjugate side groups from hormone metabolites by non-microbial action or release of glucocorticoids from lipid micelles) may also have increased fecal glucocorticoid measurements. Our findings suggest that fecal samples exposed to rainfall for one week may artificially inflate fecal glucocorticoid measurements. Thus, researchers should recognize the potential bias when collecting fecal samples exposed to rainfall. Non-fresh samples may prove useful when care is taken to address the elevation in immunoreactive glucocorticoid concentrations.