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PubMed2005

Effect of intensity of fecal pat sampling on estimates of Escherichia coli O157 prevalence.

OBJECTIVE: To evaluate site-to-site variation within fecal pats from cattle with regard to detection of Escherichia coli O157 and determine the effect on the accuracy of prevalence estimates of assay of multiple samples collected from the same fecal pat. SAMPLE POPULATION: 120 freshly voided fecal pats collected from 2 beef feedlots. Procedures-5 samples were systematically collected from each fecal pat and analyzed for E coli O157 via selective preenrichment techniques, immunomagnetic separation, and biochemical tests. Presumptive isolates were definitively identified via agglutination assays and polymerase chain reaction techniques. Best estimators of prevalence were calculated from the distribution of E coli O157-positive samples per pat. RESULTS: Of the 120 fecal pats, 96, 13, 4, 2, 3, and 2 fecal pats had 0, 1, 2, 3, 4, and 5 E coli O157-positive samples, respectively. The greatest estimate of E coli O157 prevalence (20%) was achieved when all 5 samples were assessed; this estimate represented a 2.4- fold increase in prevalence, compared with that provided via analysis of 1 sample/pat (8.2%). Compared with assessment of 5 sites/pat, the relative sensitivity of detecting an E coli O157-positive fecal pat via analysis of 1 site/pat was 40.1%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that estimates of E coli O157 prevalence derived from sampling of 1 location/pat are likely underestimates of the true prevalence of this pathogen in fecal pats (and by extension, cattle). Additional research is warranted to confirm these results in situations of high and low prevalence and across different feedlots.

Animals↗
PubMed1999

A two-step extraction method to measure fecal steroid hormones in female cynomolgus monkeys (Macaca fascicularis).

We developed a two-step extraction method for measuring fecal steroid concentrations. In the first step, distilled water was used to extract steroids from fecal samples. In the second step, a mixture of organic solvents (hexane and ether) was used to re-extract water extracts that had been transferred to a glass tube. A portion of the upper layer of the organic solvents was transferred to separate assay-tubes for measurement of estradiol (E2) or progesterone (P), and the organic solvents were evaporated in vacuo. After phosphate-buffered saline was added to each tube, commercially supplied radioimmunoassay (RIA) kits were used to determine the steroids. We demonstrated the advantages and reliability of this method by using it to assay the steroid hormone concentrations in fecal samples and serum samples collected on the same day from female cynomolgus monkeys who showed normal menstrual cycles and from monkeys who had induced hyperfunction of ovarian steroidgenesis. Different fecal samples from each monkey were used to determine the recovery rate of each steroid in water extraction from the fecal samples and the reproductivity of hormone concentrations in the fecal samples. The results demonstrate that this two-step method is simple and effective for measuring fecal steroids for monitoring the reproductive status of cynomolgus monkeys, without having to collect serum samples.

Animals↗
PubMed1986

Digoxin and metabolites in urine and feces: a fluorescence derivatization--high-performance liquid chromatographic technique.

A high-performance liquid chromatography method is described for the determination of digoxin and its metabolites digoxigenin, digoxigenin monodigitoxoside, digoxigenin bis-digitoxoside and dihydrodigoxin (20S and 20R) excreted in urine and feces. The urine sample or fecal supernatant is extracted with methylene chloride in the presence of digitoxigenin or digitoxin as internal standard. Pre-column derivatization is achieved using 1-naphthoyl chloride with subsequent separation of the derivatized compounds on either a normal- or reversed-phase system with fluorescence detection. Recoveries for digoxin and all metabolites from fecal samples were in the range 60-74%, which is comparable to that previously determined for urine samples. Standard curve data revealed linearity over a wide range of concentrations. Coefficients of variation for the analysis were less than 10% for all compounds over a range of 5-125 ng per ml urine and 10-250 ng per 200 mg feces. Peaks for digoxin and metabolites in urine and feces were obtained when human excreta were analyzed.

Chromatography, High Pressure Liquid↗
PubMed2003

Diversity, frequency, and persistence of Escherichia coli O157 strains from range cattle environments.

Genetic diversity, isolation frequency, and persistence were determined for Escherichia coli O157 strains from range cattle production environments. Over the 11-month study, analysis of 9,122 cattle fecal samples, 4,083 water source samples, and 521 wildlife fecal samples resulted in 263 isolates from 107 samples presumptively considered E. coli O157 as determined by culture and latex agglutination. Most isolates (90.1%) were confirmed to be E. coli O157 by PCR detection of intimin and Shiga toxin genes. Pulsed-field gel electrophoresis (PFGE) of XbaI-digested preparations revealed 79 unique patterns (XbaI-PFGE subtypes) from 235 typeable isolates confirmed to be E. coli O157. By analyzing up to three isolates per positive sample, we detected an average of 1.80 XbaI-PFGE subtypes per sample. Most XbaI-PFGE subtypes (54 subtypes) were identified only once, yet the seven most frequently isolated subtypes represented over one-half of the E. coli O157 isolates (124 of 235 isolates). Recurring XbaI-PFGE subtypes were recovered from samples on up to 10 sampling occasions and up to 10 months apart. Seven XbaI-PFGE subtypes were isolated from both cattle feces and water sources, and one of these also was isolated from the feces of a wild opossum (Didelphis sp.). The number of XbaI-PFGE subtypes, the variable frequency and persistence of subtypes, and the presence of identical subtypes in cattle feces, free-flowing water sources, and wildlife feces indicate that the complex molecular epidemiology of E. coli O157 previously described for confined cattle operations is also evident in extensively managed range cattle environments.

Animals↗
PubMed1987

Functional genes for cellobiose utilization in natural isolates of Escherichia coli.

The genes for utilization of cellobiose are normally cryptic in both laboratory strains and natural isolates of Escherichia coli. A survey of natural isolates of E. coli reveals that functional genes for cellobiose utilization, while rare, are present. The fraction of E. coli that utilized cellobiose ranged from less than 0.01% in human fecal samples to 7% in fecal samples obtained from horses. Samples obtained from sheep, cows, dogs, and pigs contained 0.1 to 0.5% cellobiose-positive E. coli. Neither the previously identified cel genes nor the bgl genes from E. coli K-12 were expressed during growth on cellobiose by any of the 14 naturally occurring Cel+ isolates that were tested. All of the naturally occurring Cel+ isolates possessed a cel operon, but all were deleted for the major portion of the bgl operon. The functional cel+ genes from these natural isolates differed from the mutationally activated cel+ genes obtained in earlier studies in that (i) the mutationally activated cel+ genes were temperature sensitive, while the functional genes were not, and (ii) transport of cellobiose was inducible in the strains carrying functional cel+ genes, while it was expressed constitutively in strains carrying mutationally activated genes.

Animals↗
PubMed1991

[Biological types and drug-sensitivities of multiple strains of freshly isolated methicillin-resistant Staphylococcus aureus].

Biological studies were done on numbers of methicillin-resistant strains of Staphylococcus aureus (MRSA) either clinically isolated at medical institutes nationwide and sent to us or isolated and identified by us from samples obtained in the year 1990 from patients with various infections. The results of the studies are summarized as follows. 1. The origins of the 1,047 strains used in our studies included arterial and venous blood samples (6.9%), samples from the respiratory tract (43.3%), surgical and dermatological samples (30.2%), otorhinolaryngological and ophthalmological samples (3.8%), urological samples (7.2%), fecal samples (5.1%) and others (3.6%), thus samples from the respiratory tract were the most frequent. 2. The strains were classified according to coagulase types into type II (76.4%), type III (2.1%), type IV (15.4%), type VII (5.3%) and other types (0.8%), thus type II strains were the most numerous. When classified according to their origins, type II strains were found in a significantly high frequency in respiratory tract samples, and type IV strains appeared to be present at relatively high frequencies in the surgical, dermatological, otorhinolaryngological and ophthalmological samples. Some coagulase types were found more frequently in samples from some institutes than in others, but coagulase type distributions were similar in different localities. 3. The strains were classified according to their enterotoxin types into type A (22.8%), type B (10.5%), type C (54.9%) and other types (11.7%), thus type C strains were the most frequent. Significant correlations were observed between enterotoxin type C and coagulase type II, and type A and type IV. 4. Defining strains with minimum inhibitory concentrations (MICs) of greater than or equal to 100 micrograms/ml as high MRSA and less than or equal to 50 micrograms/ml as moderate MRSA, 94.8% of coagulase type II strains were high MRSA and 78.9% of coagulase type IV strains were moderate MRSA, thus coagulase type II strains tended to be high in methicillin resistance. 5. MIC50 and MIC90 of vancomycin (VCM) against many of these strains of MRSA were 0.78 and 1.56 micrograms/ml, respectively, suggesting that VCM has also a potent antibacterial activity against recent isolates of MRSA in Japan.

Cefamandole↗
PubMed1998

Ash content of detergent fibers in feeds, digesta, and feces and its relevance in fiber digestibility calculations.

The influence of insoluble ash in diet, digesta, and fecal samples was evaluated. Ash-free values of neutral detergent fiber (NDF) and acid detergent fiber (ADF) for feed, digesta, and fecal samples were compared with uncorrected values of NDF and ADF in two digestibility experiments. Six feed samples were selected at random along with four duodenal and two fecal samples; the latter two were a part of two digestion studies. The silica composition of feed samples was low with the exception of oat hay and rice straw. Ash-free values of NDF and ADF for digesta and fecal samples were lower than uncorrected values of NDF and ADF, and silica accounted for the majority of the differences. The use of ash-free values of NDF and ADF in both digestibility experiments with dairy cows resulted in more realistic digestibility estimates and reduced the amount of variability associated with the calculation of the digestibility of NDF and ADF in the rumen and total digestive tract. The source of the insoluble ash was not identified but could have come from individual feedstuffs or the consumption of sand. Sample contamination by silica and other ash components may influence NDF and ADF composition if the fiber residues are not ashed prior to calculation of NDF and ADF.

Animal Feed↗
PubMed1992

Evaluation of two test procedures for diagnosis of giardiasis in dogs.

Two commercially available tests, an antigen-capture ELISA for use on fecal samples, and a peroral nylon string test for use in dogs, were compared with a zinc sulfate fecal concentration technique (ZSCT) for detection of giardiasis in dogs. Of 77 dogs and 164 fecal samples (from these dogs), 33 and 52, respectively were found to be Giardia-positive on the basis of results of the ZSCT. The ELISA gave false-negative results for 10 and 14% of ZSCT-positive dogs and fecal samples, respectively, and false-positive results (relative to the ZSCT test results) in 13 and 10% of ZSCT-positive dogs and fecal samples, respectively. Of the 18 string-tested dogs, 14 were positive by results of the ZSCT. Of the 4 dogs that were Giardia-negative by ZSCT, 2 were Giardia-positive by ELISA. Dogs were sedated and given water and metoclopramide to aid passage into the duodenum of the capsule containing a nylon string. Of the 21 string tests performed on the 18 dogs, only 5 strings reached the duodenum, and 0 of the 5 yielded positive results for Giardia sp. Because the string broke in 1 dog (leaving most in the gastrointestinal tract and, therefore, producing a risk of string foreign body) further string tests were not done.

Animals↗
PubMed2000

Prevalence of Giardia sp. Cryptosporidium parvum and Cryptosporidium andersoni (syn. C. muris) [correction of Cryptosporidium parvum and Cryptosporidium muris (C. andersoni)] in 109 dairy herds in five counties of southeastern New York.

A cross-sectional study was undertaken to determine the prevalence of Giardia sp. (G. duodenalis group), Cryptosporidium parvum and Cryptosporidium andersoni (C. muris) [corrected] in dairy cattle in three different age groups, and to evaluate the association of age and season with prevalence. One hundred and nine dairy farms, from a total of 212 farms, in five counties of southeastern New York volunteered to participate. On these farms, 2943 fecal samples were collected from three defined age groups. The farms were randomly assigned for sampling within the four seasons of the year. Each farm was visited once during the study period from March 1993 to June 1994 to collect fecal samples. Demographic data on the study population was collected at the time of sampling by interviewing the farm owner or manager. At collection, fecal samples were scored as diarrheic or non-diarrheic, and each condition was later related to positive or negative infection with these parasites. Fecal samples were processed using a quantitative centrifugation concentration flotation technique and enumerated using bright field and phase contrast microscopy. In this study, the overall population prevalence for Giardia sp. was 8.9%; C. parvum, 0.9%; and C. muris, 1.1%. When considering animals most at the risk of infection (those younger than 6 months of age) Giardia sp. and C. parvum was found in 20.1 and 2.4% of the animals, respectively. Giardia sp. and C. muris were found in all age groups. There was no significant seasonal pattern of infection for any of these parasites.

Age Factors↗
PubMed1975

Internal parasites of Washington cattle.

The occurrence of internal parasites in Washinton cattle was estimated. During the survey period in 1972 and 1973, more than 3,700 fecal samples from 29 herds and 4 feedlots, and viscera from 55 cattle were examined. The percentage of fecal samples with parasite eggs or oocysts from herds was as follows: oocysts, 77%; "strongyline" eggs, 44%; Nematodirus eggs, 3%; Moniezia eggs, 3%; Fasciola eggs, 2%; Trichuris eggs, 2%; and Dictyocaulus larvae, 1%. The fecal samples from feedlots contained principally strongyline eggs which varied markedly in numbers among groups of cattle in the lots. The predominant parasite in viscera was Ostertagia spp, which was recovered from all of the cattle examined. Other parasites identified in the survey were found less frequently in these examinations. Cattle in dairy and beef operations utilizing irrigated pastures were particularly prone to Fasciola infections, and more strongyline eggs were observed there than in fecal samples from other management systems. Seasonal fluctuations in numbers of strongyline and Fasciola eggs were pronounced.

Animals↗
PubMed2005

Epidemiologic features of Campylobacter infection among cats in the upper midwestern United States.

OBJECTIVE: To describe the epidemiologic features of Campylobacter infection among cats in the Minneapolis-Saint Paul metropolitan area. DESIGN: Prevalence survey. ANIMALS: 152 cats examined at 3 private veterinary clinics and an animal humane society. PROCEDURES: Fecal samples were submitted for bacterial culture for Campylobacter spp. To determine the duration of Campylobacter carriage, follow-up fecal samples were collected from cats with positive Campylobacter culture results. RESULTS: Campylobacter organisms were cultured from 37 of the 152 (24%) fecal samples. Campylobacter isolates were identified as Campylobacter upsaliensis (29 cats), Campylobacter jejuni (2), and Campylobacter coli (1); species of the remaining 5 isolates could not be determined. Campylobacter organisms were isolated from 36 of the 122 (30%) cats that were < or = 1 year old but from only 1 of the 30 (3%) cats that were > 1 year old, and shedding was more common during the summer and fall months. No association between Campylobacter shedding and clinical signs of disease was identified. For 4 of 13 cats from which follow-up fecal samples were obtained, duration of Campylobacter carriage could not be determined because Campylobacter organisms were isolated from all follow-up samples. For the remaining 9 cats, median duration of Campylobacter carriage was 44 days. CONCLUSIONS AND CLINICAL RELEVANCE: C. upsaliensis can commonly be isolated from the feces of overtly healthy kittens in the Midwest United States. Because carriage may be prolonged, veterinarians should encourage good hand hygiene among owners of cats, especially among owners with new kittens in their household.

Animals↗
PubMed1996

Evaluation for detection of Cryptosporidium oocysts in diarrheal feces of calves.

For the detection of Cryptosporidium oocysts, fecal samples were collected from 201 calves which showed diarrhea. Among the 201 samples, 29 samples (14.4%) were positive for Cryptosporidium spp. by the DMSO-modified acid-fast stain (MAFS), 23 samples (11.4%) were positive by commercial kit (Meridian Diagnostics, Cincinnati, Ohio) and 23 by the indirect immunofluorescence antibody (IFA) assay employing the monoclonal antibody (mAb C6). When tested by both IFA and MAFS, 20 fecal samples were positive for Cryptosporidium oocysts whereas 169 fecal samples were negative. If the MAFS is considered a standard method for oocyst detection, the IFA showed 69% of sensitivity and 98% of specificity. When tested by both IFA and commercial kit, 22 fecal samples were positive for Cryptosporidium oocysts while 177 samples were negative. One sample tested by IFA was found to be false negative, when compared with the results by commercial kit. The sensitivity of IFA was calculated as high as 96%; the specificity as 99% and the predictive value was also 99%. In the present study, IFA employing the mAb C6 revealed that 23 samples (11.4%) were positive among the 201 calves showing diarrhea. Of 23 IFA positive samples, 4 samples (5%) showed cryptosporidial oocysts more than 10(5) OPG. Therefore, it is concluded that the calves showing cryptosporidial oocysts more than 10(5) OPG in the feces were highly associated with clinical cryptosporidiosis.

Animals↗
PubMedDate not supplied

Fecal polymerase chain reaction with 16S ribosomal RNA primers can detect the presence of gastrointestinal Helicobacter in dogs.

Questions about pathogenesis and therapy for Helicobacter infections in dogs could be answered with a simple, noninvasive, sensitive, and specific diagnostic test. We hypothesized that a fecal polymerase chain reaction (PCR) assay would detect Helicobacter and could be useful for assessing therapeutic responses. Paired gastric biopsies and fecal samples were obtained from 39 random source dogs (group 1). Gastric biopsies from each of these dogs had histologic evidence of gastric spiral bacteria, and paired gastric tissue and fecal samples produced a 375-base pair (bp) product when amplified by PCR with Helicobacter-specific primers. Specificity of the PCR product was confirmed by detection of expected 60-, 119-, and 196-bp products following Hinfl digestion. Direct sequencing of amplicons from paired PCR products from gastric biopsy and fecal samples from 8 group I dogs showed that gastric products had the highest homologies with known gastric Helicobacter species, whereas fecal products had the highest homologies with intestinal species. Healthy mixed-breed dogs (group II; n = 8) with histologically confirmed spiral bacteria infection were treated with a 21-day course of metronidazole, amoxicillin, and famotidine. Fecal samples were collected from group II dogs twice before and within 3 days of completion of treatment. The PCR results correctly identified 15/16 pretreatment samples as positive: 1 pretreatment sample was negative. PCR results identified 8/8 posttreatment samples as Helicobacter negative. Fecal PCR is a useful test for detecting Helicobacter infection in dogs. This assay may be useful as a screening test for infection and could be used to address questions relevant to pathogenesis and therapy.

Animals↗
PubMed1999

Characterization of group C rotaviruses associated with diarrhea outbreaks in feeder pigs.

Feces and serum specimens were collected from three farms in Michigan on which approximately 50-lb (8- to 9-week-old) pigs experienced diarrhea just after placement into all-in-all-out finishing barns. The clinical signs (profuse watery diarrhea lasting about 2 weeks and no vomiting) were similar on all farms, and the morbidity rate was high (ranging from 60 to 80%) but without mortality. Eleven diarrheic fecal samples from the farms were tested for group A and C rotaviruses by immune electron microscopy (IEM) and various assays. IEM indicated that the fecal samples reacted only with antiserum against group C rotaviruses, and polyacrylamide gel electrophoresis indicated that the samples had characteristic genomic electropherotypes for group C rotavirus. Group C rotavirus was detected by cell culture immunofluorescence (CCIF) tests in nine fecal samples, but no group A rotavirus was detected by enzyme-linked immunosorbent assay or CCIF. By reverse transcription (RT)-PCR, all 11 fecal samples were positive for group C rotaviruses, with only 2 samples positive for group A rotaviruses. However, a second amplification of RT-PCR products using nested primers detected group A rotaviruses in all samples. Analysis of nucleotide and deduced amino acid sequences of the RT-PCR product (partial-length VP7) of the group C rotavirus showed 87.2 to 91% nucleotide identity and 92.6 to 95.9% amino acid identity among two strong samples from the different farms and the Cowden strain of porcine group C rotavirus. All nine convalescent-phase serum samples tested had neutralizing antibodies to the Cowden strain, and the majority of them had neutralizing antibody against group A rotaviruses (OSU or/and Gottfried strains) by fluorescent focus neutralization tests. Although group C rotaviruses have been reported as a cause of sporadic diarrhea in suckling or weanling pigs, to our knowledge, this is the first report of epidemic diarrhea outbreaks associated with group C rotavirus in older pigs.

Animals↗
PubMed1996

Isolation of poliovirus from cases of acute flaccid paralysis.

OBJECTIVE: To evaluate the frequency of isolation of polio and other viruses from fecal samples in subjects with acute flaccid paralysis in northern parts of India. SETTING: Rural and Urban Immunization Centers used for Expanded Programme on Immunization. METHODS: 219 fecal samples were collected by anal tube in 1992 from Punjab, Haryana, Himachal Pradesh and Chandigarh and processed for virus isolation. RESULTS: Of 219 fecal samples, 103 (47%) were positive for viruses in the age group of 3 mo to 5 yr. Amongst the positive samples, the predominant isolated was poliovirus type 1 (70.9%) from all the States. Non polio enteroviruses were isolated from 20.4% of fecal samples. CONCLUSION: Even though poliovirus is still the most common etiologic agent for acute flaccid paralysis in northern India, non polio enteroviruses are also emerging as important causal pathogens in this condition.

Acute Disease↗
PubMedDate not supplied

[Yersinia and infantile gastroenteritis].

Fecal samples and intestinal mucosal biopsies of 16 patients in paediatric age, with protracted diarrhoea with blood and mucus, were examined for intestinal pathogens including Yersinia. Yersinia enterocolitica 1/5/XZ and Yersinia intermedia 1/18/XZ were isolated from fecal samples of two patients. In another child, Yersinia enterocolitica, 1/5/XZ was isolated from fecal sample, while Yersinia enterocolitica 1/self-agglutinable/XZ from colon biopsy. The other intestinal pathogens were absent; Aeromonas hydrophila however was isolated from two fecal samples. The Yersinia strains isolated are not frequently found in human infections.

Adolescent↗
PubMed2003

Comparison of diagnostic techniques for Helicobacter cetorum infection in wild Atlantic bottlenose dolphins (Tursiops truncatus).

Helicobacter cetorum sp. nov. has been cultured from the stomach of Atlantic white-sided dolphins (Lagenorhynchus acutus) and the feces of Pacific white-sided (L. obliquidens) and Atlantic bottlenose (Tursiops truncatus) dolphins and a beluga whale (Delphinapterus leucas). H. cetorum has high homology to Helicobacter pylori as shown by 16S rRNA sequencing, and H. cetorum infection has been associated with gastritis and clinical signs in cetaceans. Because the prevalence of H. cetorum in wild populations is unknown, minimally invasive techniques for detecting H. cetorum were compared for 20 wild bottlenose dolphins sampled as part of a long-term health study. Fecal samples were tested for helicobacter by culture, Southern blotting, and PCR using genus-specific and H. cetorum-specific primers. An enzyme-linked immunosorbent assay (ELISA) was developed to measure H. cetorum immunoglobulin G (IgG). H. cetorum was cultured from 4 of 20 fecal samples, 7 samples were positive using Helicobacter sp. PCR, and 8 samples were positive for H. cetorum using species-specific primers. Two additional fecal samples were positive by Helicobacter sp. Southern blotting, suggesting infection with another helicobacter. All 20 sera contained high levels of IgG antibodies to H. cetorum that were significantly lowered by preabsorption of the sera with whole-cell suspensions of H. cetorum (P < 0.02). Until the specificity of the serum ELISA can be determined by testing sera from dolphins confirmed to be uninfected, PCR and Southern blot screenings of feces are the most sensitive techniques for detection of H. cetorum, and results indicate there is at least a 50% prevalence of H. cetorum infection in these dolphins.

Animals↗
PubMed1978

Coccidia of whooping cranes.

Coccidial oocysts were observed in 6 of 19 fecal samples from free-ranging whooping cranes (Grus americana) and 4 of 16 samples from captive whooping cranes. Eimeria gruis occurred in four free-ranging whooping cranes and E. reichenowi in two free-ranging and two captive whooping cranes. Fecal samples from two captive cranes contained oocysts of Isospora lacazei which was considered a spurious parasite. Oocysts of both species of Eimeria were prevalent in fecal samples collected from three free-ranging Canadian sandhill cranes (G. canadensis rowani) from whooping crane wintering grounds in Texas. These coccidia were prevalent also in fecal samples from 14 sandhill cranes (of 4 subspecies) maintained in captivity at the Patuxent Wildlife Research Center in Maryland.

Animals↗