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PubMed2000

Prevalence of enteric zoonotic organisms in cats.

OBJECTIVE: To determine prevalence of enteric zoonotic organisms in cats in north-central Colorado. DESIGN: Prospective study. SAMPLE POPULATION: Serum and fecal samples from 87 cats with diarrhea, 106 cats without diarrhea, and 12 cats for which fecal consistency was unknown. PROCEDURES: Samples were obtained from client-owned cats and cats at a humane society shelter. Serum was assayed for feline leukemia virus antigen and antibodies against feline immunodeficiency virus, IgM antibodies against Toxoplasma gondii, and IgG antibodies against T gondii and Cryptosporidium parvum. Microscopic examination of unstained feces was performed after centrifugation in a zinc sulfate solution, thin fecal smears were stained with acid fast stain and examined for C parvum, and bacteriologic culture of feces was used to detect aerobic and anaerobic bacteria. RESULTS: Enteric zoonotic organisms were detected in feces from 27 of 206 (13.1%) cats and included C parvum (5.4%), Giardia spp (2.4%). Toxocara cati (3.9%), Salmonella enterica serotype Typhimurium (1.0%), and Campylobacter jejuni (1.0%); each organism was detected in samples from cats with and without diarrhea. Although differences between groups were not significant, a higher proportion of shelter cats (18.2%) had enteric zoonotic organisms than client-owned cats (10.1%). CONCLUSIONS AND CLINICAL RELEVANCE: Enteric zoonotic organisms were detected in feces of 13.1% of cats, suggesting that cats, particularly those in homes of immunocompromised humans, should be evaluated for enteric zoonotic organisms.

Animals↗
PubMed2001

Fusobacterium prausnitzii and related species represent a dominant group within the human fecal flora.

The human gut microflora plays a key role in nutrition and health. It has been extensively studied by conventional culture techniques. However these methods are difficult, time consuming and their results not always consistent. Furthermore microscopic counts indicate that only 20 to 40% of the total flora can be cultivated. Among the predominant species of the human gut, Fusobacterium prausnitzii was reported either as one of the most frequent and numerous species or was seldom retrieved. We designed and validated a specific rRNA-targeted oligonucleotide probe, called S-*-Fprau-0645-a-A-23, to accurately detect and quantify F. prausnitzii and relatives within the human fecal microflora. The target group accounted for 5.3 +/- 3% of total bacterial 16S rRNA using dot blot hybridization (10 human fecal samples) and 16.5 +/- 7% of cells stained with Dapi using in situ hybridization (10 other human fecal samples). A specific morphology seemed to be typical and dominant: two cells forming an asymmetrical double droplet. This work showed that F. prausnitzii and phylogenetically related species represent a dominant group within the human fecal flora.

Adult↗
PubMed2002

Efficiency of fecal steroid hormone measurement for assessing reproductive function in the Hokkaido brown bear (Ursus arctos yesoensis).

The present study aimed to establish simple systems for measuring fecal steroid hormones in order to monitor the reproductive profiles of captive Hokkaido brown bears. The efficiency of fecal sample processing at the steps of dehydration and extraction and the correlation between steroid concentrations in matched fecal and blood samples were studied. Then, monthly changes in fecal estradiol-17 beta and progesterone in female bears, and testosterone in male bears were examined. The procedure was finalized as follows. Fecal samples were dried at 100 degrees C for 3 hr and extracted with diethyl ether. The diethyl ether in the extracts was evaporated and residues were reconstituted in ethanol for the assays. Hormone concentrations were quantified using enzyme immunoassays. Concentrations of progesterone and testosterone in fecal and plasma samples were correlated in the systems. The changes in fecal progesterone and testosterone concentrations were similar to those in serum concentrations of bears as reported previously. In contrast, fecal estradiol concentrations did not correlate with plasma levels probably because of the time lag in excretion. However, the changes in estradiol-17 beta concentrations in feces in the present study were similar to those reported in serum. In conclusion, fecal progesterone and testosterone assay systems appear practical for monitoring ovarian and testicular activities without immobilization, though methodological improvements and further validation may be required. For the fecal estradiol-17 beta assay, there is a need to solve the problem of excretion time lag before the system can be used in the study of reproductive physiology.

Animals↗
PubMed1995

Prevalence of enterohemorrhagic Escherichia coli O157:H7 in a survey of dairy herds.

The prevalence of Escherichia coli O157:H7 in dairy herds is poorly understood, even though young dairy animals have been reported to be a host. From February to May 1993, 662 fecal samples from 50 control herds in 14 states, and from June to August 1993, 303 fecal samples from 14 case herds in 11 states were collected for isolation of E. coli O157:H7. Case herds were those in which E. coli O157:H7 was isolated from preweaned calves in a previous U.S. Department of Agriculture study, whereas control herds from which E. coli O157:H7 had not been isolated previously were randomly selected from the same states as case herds. Among the control herds, E. coli O157:H7 was isolated from 6 of 399 calves (1.5%) that were between 24 h old and the age of weaning and from 13 of 263 calves (4.9%) that were between the ages of weaning and 4 months. Eleven of 50 control herds (22%) were positive. Among the case herds, E. coli O157:H7 was isolated from 5 of 171 calves (2.9%) that were between 24 h old and the age of weaning and from 7 of 132 calves (5.3%) that were between the ages of weaning and 4 months. Seven of 14 case herds (50%) were positive. Sixteen of 31 isolates were obtained by direct plating, with populations ranging from 10(3) to 10(5) CFU/g. Fifteen of 31 isolates were isolated by enrichment only. Nineteen of the isolates produced both verocytotoxin 1 (VT-1) and VT-2, whereas 12 produced VT-2 only.

Animal Husbandry↗
PubMed2003

Association of enteric shedding of bovine torovirus (Breda virus) and other enteropathogens with diarrhea in veal calves.

OBJECTIVE: To determine the prevalence, fecal shedding pattern, and association of bovine torovirus (BoTV) with diarrhea in veal calves at time of arrival and periodically throughout the first 35 days after their arrival on a veal farm. ANIMALS: 62 veal calves. PROCEDURE: Fecal samples collected on days 0, 4, 14, and 35 after arrival were tested for BoTV by use of ELISA and reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Paired serum samples obtained from blood collected on days 0 and 35 were analyzed for BoTV antibodies with a hemagglutination inhibition assay. Fecal samples were also screened for other enteric pathogens, including rotavirus, coronavirus, and Cryptosporidium spp. RESULTS: Fecal shedding of BoTV was detected in 15 of 62 (24%) calves by use of ELISA and RT-PCR assay, with peak shedding on day 4. A significant independent association between BoTV shedding and diarrhea was observed. In addition, calves shedding > or = 2 enteric pathogens were more likely to have diarrhea than calves shedding < or = 1 pathogen. Calves that were seronegative or had low antibody titers against BoTV (< or = 1:10 hemagglutination inhibition units) at arrival seroconverted to BoTV (> 4-fold increase in titer); these calves were more likely to shed virus than calves that were seropositive against BoTV at arrival. CONCLUSIONS AND CLINICAL RELEVANCE: Shedding of BoTV was strongly associated with diarrhea in neonatal veal calves during the first week after arrival at the farm. These data provide evidence that BoTV is an important pathogen of neonatal veal calves.

Animals↗
PubMed2003

Efficacy of Giardia vaccination in the treatment of giardiasis in cats.

OBJECTIVE: To assess efficacy of Giardia vaccination as a treatment for giardiasis in experimentally infected cats. DESIGN: Original study. ANIMALS: 16 young-adult cats. PROCEDURE: Cats were experimentally infected by orogastric administration of Giardia cysts. On weeks 4, 6, and 10, cats in the treatment group (n = 8) were given Giardia vaccine SC. For the first 28 weeks after infection, 3 fecal samples from each cat were examined weekly for Giardia cysts, and cyst numbers were counted. Fecal consistency was scored daily for the duration of the study. Results from vaccinated and unvaccinated cats were compared by logistic regression. RESULTS: All cats became infected and were shedding Giardia cysts by the end of week 2. Throughout the study, diarrhea was rare and was mild and transient when it did occur. By week 28, 5 of 8 vaccinated cats and 7 of 8 control cats had patent Giardia infections. Magnitude of infection, based on number of fecal samples with cysts and number of cysts per sample, decreased progressively in both groups over time. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of 3 doses of a Giardia vaccine did not completely eliminate the organism from experimentally infected cats in the study period. Since clinical signs were minimal in both groups of cats, it could not be determined whether vaccination lessened severity of clinical disease. Results may have been negatively influenced by the large inoculation dose. Whether Giardia vaccination is an effective treatment for giardiasis in naturally infected cats remains to be determined.

Animals↗
PubMed2002

Prevalence of Salmonella and E. coli, and their resistance to antimicrobial agents, in farming communities in northern Thailand.

A cross-sectional pilot study was conducted in Chiang Mai, Thailand, to determine the prevalence of Salmonella and Escherichia coli in swine, broiler chickens and human workers from farms and abattoirs in northern Thailand, and compare their antimicrobial resistance profiles. Fecal samples and cloacal swabs were collected from 150 swine and 150 chickens at the farm. Fecal samples from swine, cloacal swabs from chickens, and carcass swabs from both animals were collected from 100 swine and 100 chickens at the abattoir. Stool samples were collected from 15 swine farm workers and seven chicken farm workers. Primary isolation and identification of Salmonella and E. coli were conducted using standard methods. In vitro susceptibility testing of Salmonella and E. coli was conducted using the broth microdilution method, based on the United States National Committee for Clinical Laboratory Standards (NCCLS) guidelines. The prevalence of Salmonella from swine and chicken samples ranged from 2% to 25%. The prevalence of E. coli in chickens and swine ranged from 36.8% to 47.6%. In humans, the prevalence of Salmonella was 15%, and the prevalence of E. coli ranged from 51% to 53%. Resistance in Salmonella was found for tetracycline (84.7%), nalidixic acid (27.1%), florfenicol (18.6%), ampicillin (13.6%), and ceftiofur (3.4%), and in E. coli for tetracycline (91.5%), nalidixic acid (67.4%), ampicillin (61.6%), florfenicol (51.8%), enrofloxacin (28.7%), ciprofloxacin (12.5%), ceftiofur (4.9%) and ceftriaxone (1.5%).

Animals↗
PubMed1979

Paratuberculosis in cattle and free-living exotic deer.

Paratuberculosis was studied among dairy cows and exotic deer that shared grazing areas at Point Reyes National Seashore, California. Of the 10 dairy herds tested, 5 (50%) were infected with Mycobacterium paratuberculosis (based on results of fecal culture). Mycobacterium paratuberculosis was cultured from 9 (8.7%) of the 103 bovine fecal samples and from 4 (3.9%) of the 103 bovine rectal mucosa scapings tested. Of 89 fecal samples from 52 axis deer (Axis axis) and 37 fallow deer (Dama dama), 5 (9.6%) and 3 (8.1%), respectively, contained M paratuberculosis. Culture of intestinal necropsy samples from the same deer indicated that 3 (5.8%) of the axis deer and 2 (5.4%) of the fallow deer were infected with M paratuberculosis. The cows were tested for serum antibodies by the complement-fixation test and by radioimmunoassay. Of 95 sera tested by complement fixation, 15 (15.8%) were positive, as were 15 (14.7%) of 102 sera tested by radioimmunoassay. Culture results and serologic test results were compared on a herd basis.

Animals↗
PubMed1980

Hemagglutination by canine parvovirus: serologic studies and diagnostic applications.

Conditions for canine parvoviral hemagglutination (HA) and hemagglutination-inhibition (HI) reactions were defined. The HA phenomena were used to differentiate canine parvovirus (CPV) from feline panleukopenia virus (FPV), mink enteritis virus (MEV), and minute virus of canines. Serologic comparisons of the CPV, FPV, and MEV by HA-HI and serum-neutralization tests indicated that CPV, FPV, and MEV were antigenically similar but were different from minute virus of canines. Diagnostic application of HA tests to fecal samples from acute cases of enteritis was discussed. Combinating HA tests with HI tests on fecal samples provided a rapid and specific diagnostic method for CPV infection. Secular seroprevalence studies indicated the emergence of CPV infeciton in the United States dog population-at-large in 1978.

Animals↗
PubMed1993

Use of fenbendazole for treatment of Crenosoma vulpis infection in a dog.

An 8-month-old Labrador Retriever was examined because of a 1-month history of productive coughing unresponsive to ampicillin treatment. Larvae of Crenosoma vulpis were found in fecal samples examined by zinc sulfate centrifugation and Baermann technique. Physical examination abnormalities or larvae in fecal samples were not detected 6 weeks after treatment with prednisone (1 mg/kg, PO, q 24 h, for 7 days, then 0.5 mg/kg, PO, q 48 h, for 8 days) and fenbendazole granules (50 mg/kg, PO, q 24 h, for 3 days). This report suggests that fenbendazole may be effective for treating Crenosoma vulpis infection in dogs.

Animals↗
PubMed1997

Porcine reproductive and respiratory syndrome virus: a persistent infection.

Persistent infection with porcine reproductive and respiratory syndrome virus (PRRSV) was shown in experimentally infected pigs by isolation of virus from oropharyngeal samples for up to 157 days after challenge. Four 4 week old, conventional, PRRSV antibody-negative pigs were intranasally inoculated with PRRSV (ATCC VR-2402). Serum samples were collected every 2 to 3 days until day 42 post inoculation (PI), then approximately every 14 days until day 213 PI. Fecal samples were collected at the time of serum collection through day 35 PI. Oropharyngeal samples were collected at the time of serum collection from 56 to 213 days PI by scraping the oropharyngeal area with a sterile spoon, especially targeting the palatine tonsil. Turbinate, tonsil, lung, parotid salivary gland, spleen, lymph nodes and serum were collected postmortem on day 220 PI. Virus isolation (VI) on porcine alveolar macrophage cultures was attempted on all serum, fecal and oropharyngeal samples, as well as tissues collected postmortem. Postmortem tonsil tissues and selected fecal samples were also assayed for the presence of PRRSV RNA by the polymerase chain reaction (PCR). Serum antibody titers were determined by IFA, ELISA and SVN. Virus was isolated from all serum samples collected on days 2 to 11 PI and intermittently for up to 23 days in two pigs. No PRRSV was isolated from fecal samples, but 3 of 24 samples were PCR positive, suggesting the presence of inactivated virus. Oropharyngeal samples from each pig were VI positive 1 or more times between 56 and 157 days PI. Oropharyngeal samples from 3 of 4 pigs were VI positive on days 56, 70 and 84 PI. Virus was isolated from one pig on day 157 PI, 134 days after the last isolation of virus from serum from this animal. Virus was isolated from oropharyngeal samples for several weeks after the maximum serum antibody response, as measured by IFA, ELISA and SVN tests. All tissues collected postmortem were VI negative and postmortem tonsil samples were also negative by PCR. An important element in the transmission of PRRSV is the duration of virus shedding. The results of this study provided direct evidence of persistent PRRSV infection and explain field observations of long-term herd infection and transmission via purchase of clinically normal, but PRRSV infected, animals. Effective prevention and control strategies will need to be developed in the context of these results.

Animals↗
PubMed1991

Pregnancy determination in uncaptured feral horses by means of fecal steroid conjugates.

This study was carried out to develop an accurate, rapid and inexpensive method for diagnosing pregnancy in uncaptured feral horses by analysis of fecal steroid metabolites and to compare the accuracy of this method with diagnosis by urinary estrone conjugates (E(1)C). Paired urine and fecal samples were collected from 40 sexually mature feral mares during August and October. Urine samples were extracted directly from the soil and analyzed by enzymeimmunoassay (EIA) for E(1)C. Water extracts of fecal samples were assayed by EIA for E(1)C and nonspecific progesterone metabolites (iPdG). Urinary E(1)C, fecal E(1)C and fecal iPdG concentrations for seven mares which produced foals were 3.9 +/- 1.3 (SEM) mug/mg creatinine, 4.2 +/- 0.8 ng/g feces and 1.411 +/- 569.6 ng/g feces, respectively. Urinary E(1)C and fecal E(1)C and iPdG concentrations for the 33 mares which did not produce foals were 0.1 +/- 0.0 mug/mg creatinine and 0.5 +/- 0.1 and 32.8 +/- 4.5 ng/g feces, respectively. These differed (P < 0.01) from values in mares which produced foals.

Journal Article↗
PubMed2005

Identification of urolithin a as a metabolite produced by human colon microflora from ellagic acid and related compounds.

Dietary ellagic acid and related polyphenols are metabolized in humans to dibenzopyran-6-one derivatives, and the microbial origin of these metabolites has been suggested. However, this has not been demonstrated so far. Fecal samples donated by six volunteers were incubated under anaerobic conditions, and aliquots were used to evaluate the fecal metabolism of ellagic acid, the ellagitannin punicalagin, and an ellagitannin rich extract from walnuts. The isoflavone daidzein was also incubated with the same fecal samples to follow the production of the microbial metabolites previously reported (dihydrogenistein, O-demethylangolensin, and equol) as a positive control of the system and to evaluate similarities between isoflavone and ellagic acid fecal flora metabolism. After fermentation the metabolite "urolithin A" (3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one) was produced from ellagic acid, punicalagin, and the ellagitannin extract in all the fecal cultures from different volunteers, but with very different production rates and concentrations. This large variability in the concentration of metabolite and kinetics of metabolite production is consistent with the large variability found in the excretion of these metabolites in urine in vivo after human consumption of ellagitannins, and with differences in the composition of the fecal microflora. No correlation between isoflavone and ellagic acid metabolism by fecal microflora was observed. The present study confirms the microbial origin of the recently reported in vivo generated hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives in humans and is a further step in the study of the bioavailability and metabolism of ellagic acid and ellagitannins.

Bacteria↗
PubMed1990

[Adult diarrhea rotavirus in Jilin, China].

In May 1986, an outbreak of epidemic acute diarrhea occurred in one city and three counties in Yanbian area. The diarrheal cases were seen in all age groups, the majority of the cases were seen in adult age group. No bacterial pathogens were isolated in 22 fecal samples examined, however rotavirus like particles of 52-68 nm in diameter were found in 50% (11/22) of fecal samples by the immunoelectron microscopy using convalescent sera. Examination by the adult diarrhea rotavirus (ADRV) ELISA kit showed positive reaction in 86% (6/7) of fecal extracts, however in all 22 fecal extracts examination using ELISA kit for detection of group A rotavirus showed negative reaction. The PAGE patterns of viral RNAs similar to those of ADRV were seen in 13 out of 22 fecal extracts. The detection rate by PAGE analysis was 59%. Based on the above data, the etiological agent of the epidemic acute diarrhea, which occurred in Yanbian area, was identified to be ADRV.

Acute Disease↗
PubMed1987

Gastrointestinal nematode parasites in Saskatchewan cattle: egg count distributions in beef animals.

Fecal samples were collected systematically in the spring from cows and yearlings entering, and in the fall from calves leaving, five community pastures in central Saskatchewan. Fecal samples were also collected systematically in the spring from cows entering, and in the fall from calves leaving, an experimental rotational grazing system. Samples were collected from 1398 animals and were examined by a quantitative fecal flotation technique with a sensitivity of ten eggs per gram. Distributions of nematode egg counts for 11 of the 12 cattle populations sampled were well described by a series of negative binomial distributions. A common value of k (a measure of aggregation) could be fitted to the counts from animals of all ages in four of the five community pastures, as well as to the counts from the cows from the fifth community pasture and from the rotational grazing system. A second value of k could be fitted to the counts from the calves from these two pasture. In addition, in three of the community pastures animals in the different age groups had the same mean count.

Animals↗
PubMed2003

Transmission of Salmonella enterica serotype typhimurium DT104 to infants through mother's breast milk.

This study documents the first reported transmission of Salmonella enterica serotype Typhimurium definitive type 104 (DT104) to premature fraternal twins via their mother's breast milk. When premature twin neonates developed severe enteritis in the neonatal intensive care unit (NICU), stool samples and the mother's breast milk were cultured for the presence of Salmonella. Antibacterial susceptibility patterns were determined. Semiquantitative organism abundance data were retrospectively gathered on 54 stored breast milk samples collected on 34 different days using a rapid, real-time polymerase chain reaction (PCR) methodology (LightCycler PCR). Fecal samples from other infants in the NICU at that time were also tested. Pulsed-field gel electrophoresis (PFGE) was used to assess the genetic composition of the isolated organisms. The twins' neonatal stools and mother's breast milk cultures revealed a resistance pattern (R-type) to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. LightCycler PCR analysis of sequential breast milk samples confirmed this to be the likely source of transmission. In the subsequent outbreak investigation, none of the NICU surveillance fecal samples proved positive for this organism. The genetic composition of organisms isolated from the maternal breast milk was indistinguishable from those isolated from neonatal specimens as determined by PFGE. Antibiotic susceptibility tests coupled with PFGE patterns suggested that these Salmonella isolates were DT104. Because the prevalence of DT104 infections is rising in the United States, neonatologists should be aware of breast milk as a potential mode of transmission.

Diseases in Twins↗
PubMed2005

Detection of Lawsonia intracellularis using immunomagnetic beads and ATP bioluminescence.

Lawsonia intracellularis is an obligate intracellular pathogenic bacterium that causes proliferative enteropathy in various animals. The detection of L. intracellularis in clinical and environmental samples is necessary for the diagnosis of infection and epidemiological investigations. For the detection of L. intracellularis in fecal samples, we have developed an immunological method using immunomagnetic separation and ATP bioluminescence. Magnetic beads were coated with an anti-Lawsonia surface antigen (LsaA) antibody in order to capture the L. intracellularis in fecal samples from infected rabbits and the bacteria captured by the immunomagnetic beads were assayed by means of ATP bioluminescence. Our results showed that L. intracelluraris was detected by immunomagnetic separation of bacteria-holding magnetic beads and ATP-based bioluminescence, suggesting that our methods could be useful for the diagnosis of proliferative enteropathy.

Adenosine Triphosphate↗