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PubMed2004

Evaluation of adrenal function in serum and feces of Steller sea lions (Eumetopias jubatus): influences of molt, gender, sample storage, and age on glucocorticoid metabolism.

Fecal corticosterone concentrations, measured via radioimmunoassay (RIA), were validated as a method to monitor adrenal function in Steller sea lion physiology. Quantification of adrenal response to an acute stressor and relevance of data produced by developed methodologies was determined through physiological challenge with exogenous administration of adrenocorticotropic hormone (ACTH) to captive adult, reproductively intact, Steller sea lions of both sexes (n=3, 1 male, 2 female) during seasonal molt. Following ACTH administration, serial blood and fecal samples were collected and analyzed by RIA to determine adrenal response. Storage regimens and weather exposure were examined to establish external impact on fecal corticosterone concentrations. High-pressure liquid chromatography (HPLC) of both serum and feces of Steller sea lions was employed to explore potential gender-based differences extant in either sample media. ACTH challenges produced >3-fold increases in serum cortisol concentrations which were reflected in >18-fold increases in fecal corticosterone concentrations post-injection at 3.25 and 32 h, respectively, and fecal corticosterone concentrations returned to baseline 52 h post-injection. Neither outdoor exposure to weather nor variation in duration and temperature of freezer storage impacted fecal corticosterone concentrations. HPLC of individual fecal samples produced eluate immunoreactivity profiles that differed consistently with both sex and age class. Techniques developed herein effectively detected physiologically relevant corticosterone data in Steller sea lion feces, unaffected by conditions likely to be encountered with field collection samples. Additionally, results quantify an acute response to ACTH and provide methodology for examining chronically heightened adrenal activity in Steller sea lions.

Adrenal Glands↗
PubMed2006

Herd-level risk factors for Salmonella enterica subsp. enterica in U.S. market pigs.

Midwest U.S. herds (n=63) were studied to identify risk factors for harboring Salmonella enterica among slaughter-weight pigs. Samples collected on farms (feces) and at slaughter (distal colonic content, cecal content and ileocolic lymph nodes) were cultured using conventional means. Approximately 15 pigs were studied per herd, for a total of 3754 samples. The proportion of pigs positive in one or more samples was calculated for each herd. Herd characteristics were described by a combination of interview and written survey. Logistic regression was used to detect relationships between the detection of Salmonella and potential herd-level risk factors. The mean individual pig prevalence was 5% for feces, 4% for distal colonic content, 15% for ileocolic lymph nodes, and 17% for cecal contents. One or more Salmonella isolates were detected in at least one sample type in every herd. The five most common serovars were S. Agona, S. Derby, S. Schwarzengrund, S. Typhimurium and S. Senftenberg, with 25 additional serovars detected. Salmonella prevalence estimates were positively correlated among all samples except distal colonic content and ileocolic lymph nodes. Pigs with culture positive fecal samples were at increased odds of being detected positive for each of the slaughter-collected samples examined, namely distal colonic content (OR=30.5), ileocolic lymph nodes (OR=12.9) and cecal content (OR=23.2). Herds with positive fecal sample(s) had increased odds of having positive cecal content (OR>1.5), distal colonic content (OR=15.3) and ileocolic lymph nodes (OR=12.7). Pigs from herds with at least some bowl drinkers had eight-fold higher odds of testing Salmonella positive than did pigs from herds with only nipple drinkers. Pigs from herds with only dry feeders had five-fold higher odds of testing Salmonella positive when compared with pigs from herds with combinations of wet/dry style feeders. Interventions at these two points should be considered when designing growing pig facilities to reduce Salmonella shedding.

Animal Feed↗
PubMed2004

Prevalence of Giardia duodenalis genotypes in pre-weaned dairy calves.

To determine the prevalence of Giardia genotypes in pre-weaned dairy calves, fecal samples were collected from a minimum of 18, 1-7-week-old dairy calves per farm on two farms each in the states of Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Samples cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and gene sequence analysis. Prevalence by PCR ranged from 9% on a farm in Pennsylvania to 93% on a farm in Vermont, with an average prevalence for 407 calves on 14 farms of 40%. Gene sequence analysis of the TPI, beta-giardin and 16S rRNA genes revealed 85% of the positive samples to be Assemblage E, while 15% were Assemblage A, although the percentages of these genotypes varied greatly from farm to farm. Some farms had no Assemblage A Giardia. Thus, while a majority of the calves were infected with a genotype that is not known to be infectious for humans, calves on 7 of 14 farms did harbor Assemblage A Giardia. Calves should be considered as a potential source of human infectious cysts in the environment.

Animals↗
PubMed1992

Development of a biotin-streptavidin-enhanced enzyme-linked immunosorbent assay which uses monoclonal antibodies for detection of group C rotaviruses.

A biotin-streptavidin-enhanced enzyme-linked immunosorbent assay (ELISA) which uses monoclonal antibodies (MAbs) for the detection of group C rotaviruses was developed. An assay in which plates were coated with three pooled MAbs and biotinylated polyclonal immunoglobulin G (IgG) (polyclonal antibody [PAb]) was used as the detector (MAb capture-PAb detector) was found to be the most sensitive and specific of the assays when it was compared with assays in which plates were coated with polyclonal antiserum and detection was done with either biotinylated polyclonal antiserum (PAb capture-PAb detector) or biotinylated pooled MAbs (PAb capture-MAb detector). The MAb capture-PAb detector ELISA detected 83% of samples confirmed to be positive for group C rotaviruses, whereas the PAb capture-PAb detector assay detected 63% of positive samples and the PAb capture-MAb detector assay detected 65% of positive samples. All three procedures detected both of the bovine and the two human group C rotaviruses, but none of the three procedures detected fecal samples containing group A and B rotaviruses or fecal samples negative for group C rotaviruses used in this study. The sensitivity of the MAb capture-PAb detector ELISA was determined by serially diluting fecal group C rotaviruses; antigens were detected in maximal positive dilution ranges of 1:1,000 to 1:3,000 for the samples tested. On the basis of the cell culture immunofluorescence assay infectivity titer of semipurified cell culture-passaged Cowden group C rotavirus, the sensitivity of the MAb capture-PAb detection ELISA for detection of homologous group C rotavirus was 53 fluorescent focus units per ml. Epitope mapping by use of the biotinylated MAbs in competition assay suggested that our MAbs may bind to three different but overlapping epitopes. These results suggest that the MAb capture-PAb detector ELISA can be used to study the epidemiology of group C rotaviruses in humans and animals.

Animals↗
PubMed2002

Spread of hepatitis E virus among different-aged pigs: two-year survey in Taiwan.

Swine are reservoirs of hepatitis E virus (HEV). In this study, a 2-year survey of HEV in feces and sera of swine was conducted to determine if: 1) HEV has circulated among pigs for some time in Taiwan; 2) the spread of HEV among different-aged pigs; and 3) there exists HEV strains possibly imported through trading. From 1998-2000, 521 serum samples and 54 fecal specimens from pigs were examined by reverse transcription polymerase chain reaction. None of the 11 pigs in suckling stage (< 2 months) were serum HEV RNA positive. The highest viremia rate (4.5%) was in pigs of 2 months age, followed by 1.2% and 1.8% in pigs of growing (3-4 months) and finishing stages (5-6 months), and none in pigs older than 6 months. Viremia showed little variation in different years and areas. None of the 20 fecal samples from pigs in suckling stage were HEV RNA positive, whereas 9% of the 34 samples from pigs in growing or finishing stages were positive. Most swine HEV isolates in Taiwan clustered within the genotype 4, whereas the three HEV isolates cloned from pigs imported recently from the U.S. belonged to the genotype 3 HEV in the U.S. The results suggest that HEV may infect pigs at an early growing stage and spread unnoticed among pigs and possibly across countries through trading.

Age Factors↗
PubMed2005

Rapid PCR detection of enterohemorrhagic Escherichia coli (EHEC) in bovine food products and feces.

Although Escherichia coli (E. coli) O157:H7 is a major cause of foodborne illness, other types of E. coli can also cause illness. E. coli that possess the eae gene for attachment and effacing have the potential to cause disease. Many real-time, molecular-based assays have been developed to detect Enterohemorrhagic E. coli (EHEC) including E. coli O157:H7. However, no assay currently exists to detect the eae gene present in E. coli O157:H7 and other EHEC strains with a confirmed positive or negative result in less than 12 h. Raw beef food products (raw ground beef and raw boneless beef) at 25 and 375 g samples and bovine fecal samples at 2 g were inoculated with 10(1), 10(3), 10(4), and 10(5) organisms of E. coli O157:H7 to test the sensitivity of this assay. Fourteen different foodborne bacteria (including E. coli O157:H7) and 19 various E. coli strains, obtained from the United States Department of Agriculture-Agricultural Research Service (USDA-ARS) were tested for specificity. E. coli O157:H7 was detected at the level of 10(1) organisms in both 25 and 375 g samples of raw ground and raw boneless beef products as well as 2 g samples of bovine feces after pre-enrichment and concentration. None of the 14 foodborne bacteria screened for cross-reactivity was detected. All USDA E. coli strains confirmed to contain the eae gene were detected.

Animals↗
PubMed2002

Noninvasive colorectal cancer screening.

Abnormal mucin with the STn epitope is produced by colorectal cancer cells and is immunologically distinguishable from normal colonic mucin. Herein we describe a technique of detecting abnormal mucin in fecal samples as a method of screening for colorectal neoplasia. Soluble glycoproteins from fecal samples of patients with symptoms of bowel disease and asymptomatic volunteer subjects were isolated by centrifugation and ethanol precipitation. The protein content of the soluble fraction was normalized and tested by immunoassay (slot dot). Anti-COTA monoclonal antibody SP-21, which reacts with STn epitope, was applied in the reaction, and the optical density of each slot dot was determined by imaging densitometer. Quantitative values of samples were determined from the standard curve obtained with highly purified COTA. COTA values > 15 microg/ml were considered positive for neoplasia. Results indicated that 5/6 colon cancers, 6/22 adenomas, 1/8 colitis cases, and 2/58 normal patients were positive in the test. The pilot study revealed that COTA assay is sensitive and more specific than Hemoccult screening for colorectal neoplasia.

Antibodies, Monoclonal↗
PubMed1996

Fecal short-chain fatty acids in patients with diarrhea-predominant irritable bowel syndrome: in vitro studies of carbohydrate fermentation.

Colonic bacterial production of short-chain fatty acids (SCFA) plays an important role in the salvage of unabsorbed carbohydrate and in colonic absorption of electrolytes and water. The objective of this study was to determine whether patients with diarrhea-predominant irritable bowel syndrome (DP-IBS) have a different pattern and rate of fermentation of carbohydrate and fiber to SCFA compared with controls. Fecal homogenates from 10 patients with DP-IBS and 10 age-matched controls were studied. SCFA were measured by gas chromatography in baseline fecal samples and in fecal homogenates in an in vitro anaerobic fermentation system after incubation with no additional substrate, lactulose, potato starch, citrus pectin, and hemicellulose over a 24-hour period. Net SCFA production rates were calculated for the first 6 h of the incubation period. Patients with DP-IBS had a consistently different pattern of less total SCFA, a lower percentage of acetate (p < 0.05), and a higher proportion of n-butyrate (p < 0.05) than controls. In stool homogenates from both controls and DP-IBS patients, lactulose fermentation resulted in the highest rate of SCFA production followed by pectin, starch, and hemicellulose. However, at all time points, the fecal homogenates from controls generated a higher concentration of total SCFA, acetate, and propionate with all substrates tested. SCFA production rates were higher in controls incubated with lactulose, starch, and hemicellulose. The fecal SCFA profile of patients with DP-IBS is characterized by lower concentrations of total SCFA, acetate, and propionate and a higher concentration and percentage of n-butyrate. Fecal flora from these patients produced less SCFA in an in vitro fermentation system in response to incubations with various carbohydrates and fibers. Differences in SCFA production by colonic bacterial flora in patients with DP-IBS may be related to the development of gastrointestinal symptoms.

Acetates↗
PubMed2004

Effect of tetracycline on transfer and establishment of the tetracycline-inducible conjugative transposon Tn916 in the guts of gnotobiotic rats.

We have investigated the transfer of Tn916 among strains of Enterococcus faecalis OG1 colonizing in the intestines of gnotobiotic rats. This animal model allows a low limit of detection and efficient colonization of the chosen bacteria. The animals continuously received tetracycline in drinking water. A tetracycline-sensitive recipient strain was allowed to colonize the animals before the resistant donor was introduced. The numbers of donors, recipients, and transconjugants in fecal samples and intestinal segments were estimated. The bioavailable amounts of tetracycline in fecal samples and intestinal segments were monitored by using bacterial biosensors carrying a transcriptional fusion of a tetracycline-regulated promoter and a lacZ reporter gene. Chromosomal locations of Tn916 in transconjugants isolated either from the same animal or from different animals were compared by Southern blot analysis. Our results indicated that selection for the resistant phenotype was the major factor causing higher numbers of transconjugants in the presence of tetracycline. Tetracycline-sensitive E. faecalis cells colonized the intestine even when the concentrations of tetracycline in feces and intestinal luminal contents exceeded growth-inhibitory concentrations. This suggests the existence of tetracycline-depleted microhabitats in the intestinal environment.

Animals↗
PubMed2006

Prevalence of hepatitis A virus infection in Goiânia, Goiás, Brazil, by molecular and serological procedures, 1995-2002.

In this study, a total of 865 serum samples were collected between 1995 and 2002 from individuals living in Goiânia, Central Brazil, and clinically suspected of hepatitis. After exclusion of 162 samples which were positive for hepatitis B virus or hepatitis C virus, 703 samples were tested for anti-hepatitis A virus (anti-HAV) IgM antibodies by enzyme immunoassay. In addition, 588 of these samples and 22 fecal samples were analyzed by reverse transcription-nested PCR for HAV RNA detection, with positivity indices of 13.1% (77/588) and 54.5% (12/22), respectively. A similar index of viral RNA detection in anti-HAV-IgM positive or negative samples was observed in serum samples. HAV infection is a public health problem worldwide and this study underscores the extent of HAV circulation in our region.

Adolescent↗
PubMed2001

Suspected Clostridium difficile-associated diarrhea in two cats.

Two adult cats from the same household developed acute diarrhea. Clostridium difficile toxins were detected in the feces of both cats, whereas other recognized causes of diarrhea were not identified. Supportive medical treatment and metronidazole were administered and both cats responded well. A fecal sample obtained from 1 of the affected cats after treatment and a fecal sample obtained from a clinically normal cat in the household did not contain C difficile toxins. The role of C difficile in enteric disease in cats has not been extensively studied and is unclear; however, our findings suggest that toxigenic strains of C difficile may cause diarrhea in cats.

Animals↗
PubMed1982

Cryptosporidiosis of man and calf: a case report and results of experimental infections in mice and rats.

Cryptosporidiosis is reported in a healthy 25-year-old male. Clinical symptoms include 1 day of nausea and low-grade fever and 9 days of diarrhea, followed by 3 days of constipation. Oocysts of Cryptosporidium sp. were present in sugar flotations of the first fecal sample collected 56 hours after onset of the symptoms and in daily fecal samples collected through day 12 of the illness. Oocysts of the human isolate of Cryptosporidium sp. were morphologically indistinguishable from those obtained from naturally and experimentally infected calves. After 1 week of sporulation at room temperature, oocysts from the human and from calves contained four sporozoites and a spherical residuum. When inoculated orally, sporulated Cryptosporidium sp. oocysts of human and of calf origin produced indistinguishable infections in suckling mice and rats and in adult mice.

Adult↗
PubMed2003

Molecular characterization of Escherichia coli O157 contamination routes in a cattle slaughterhouse.

In a cattle slaughterhouse, sampling was performed over a 1-week period to examine the prevalence and possible contamination routes of Escherichia coli O157. Each sampling day, swab samples were collected from the slaughterhouse environment before onset of slaughter, from the slaughterline, and from 20 successively slaughtered animals. Isolation of E. coli O157 consisted of a 6-hour enrichment followed by immunomagnetic separation and selective plating. From the 394 samples taken, 84 (21%) were positive for E. coli O157. Pulsed-field gel electrophoresis (PFGE) of collected isolates produced 26 different profiles, from which 5 PFGE profiles carried two or more Stx genes. The combination of PFGE profiles and Stx types resulted in 32 different E. coli O157 types. E. coli O157 was found in the slaughterhouse environment before the onset of slaughter. The first two sampling days, feces and carcasses were found negative. On the third sampling day, five fecal samples and four carcasses from animals negative in the feces were positive. Hide of the anal region and the shoulder were found positive every sampling day. The shoulder hide was more than twice as contaminated as the anal region hide. Typing of different isolates from a sample showed that frequently different E. coli O157 types were presented. On sampling days 1 and 2, types present in the environment and on the hides of the slaughtered animals differed. On the third sampling day, two dominant types were found in the environment (even before the onset of slaughter), as well as on the hides, feces, and carcasses. Although examined animals originated from different farms, one (two on day 3) dominant E. coli O157 type was present on their hides each sampling day. These data indicated that (i) the progress of contamination can differ from day to day within a slaughterhouse and (ii) contact between animals after the departure from the farm can have a large effect on the spread of E. coli O157 hide contamination.

Abattoirs↗
PubMed2005

Escherichia coli O157:H7, Campylobacter jejuni, and Salmonella Prevalence in cull dairy cows marketed in northeastern Ohio.

Preharvest management factors are predicted to impact the prevalence of foodborne pathogens in cattle sent to slaughter. We simultaneously examined the prevalence and antibiotic resistance patterns of Campylobacter jejuni, Salmonella, and Escherichia coli O157:H7 isolated from cull dairy cattle at two livestock auctions in northeastern Ohio. Between April and September 2002, a total of 1,026 fecal samples were collected. C. jejuni was isolated from 48 of 686 (7%) fecal samples, Salmonella was isolated from 39 of 585 (6.7%) samples, and E. coli O157:H7 was isolated from 21 of 1,026 (2.1%) samples. Of the 585 samples tested for all three pathogens, at least one pathogen was identified in 86 of 585 (15%) samples. One sample was positive for both E. coli O157:H7 and C. jejuni, and five samples yielded both C. jejuni and Salmonella. Size of herd of origin could be traced for 75 to 85% of samples collected. Salmonella was isolated at higher frequencies from herds larger than 60 cattle than from smaller herds (9.0 versus 3.5%, P = 0.02). In contrast, size of herd of origin did not significantly affect the E. coli O157:H7 and C. jejuni prevalence. Approximately 90% of Salmonella and E. coli O157:H7 isolates were pansensitive to a panel of 16 antibiotics. Thirty-six percent of C. jejuni isolates were resistant to tetracycline. In this study, antibiotic resistance among the foodborne pathogens isolated from cull diary cattle was rare. Although size of dairy herd of origin was positively associated with Salmonella prevalence, herd size was not strongly associated with E. coli O157:H7 and C. jejuni prevalence in market dairy cattle. These results can be used to assess the food safety risks associated with the slaughter of cull dairy cattle.

Abattoirs↗
PubMed2005

Relationships among fecal lungworm loads, fecal glucocorticoid metabolites, and lamb recruitment in free-ranging Rocky Mountain bighorn sheep.

Most wild Rocky Mountain big-horn sheep (Ovis canadensis canadensis) in northern latitudes are infected with lungworms. Indirect effects of lungworms on bighorn sheep are unknown, but high pulmonary burdens might increase stress (i.e., elevated glucocorticoid levels), and chronic stress could in turn decrease fitness. We hypothesized that high lungworm burdens in Rocky Mountain bighorn ewes increase stress, thereby increasing lamb mortality. To test our hypothesis, one subherd of bighorn sheep in Custer State Park, South Dakota was provided a free-choice loose mineral mix containing the anthelmintic fenbendazole every six weeks from March 1999 to August 2000 to eliminate lungworms; another subherd served as the control. Daily, individually marked ewes were located telemetrically from the ground and uniquely marked animals were observed until they defecated. After the herd moved from the area, fecal samples were collected and stored at -23 C. A consistent number of samples per season per herd (x-=16.56+/-3.99 samples) were collected. Fecal larval lungworm levels (LPG) in the treatment subherd were lower than levels in the control subherd; however, there was no difference in fecal glucocorticoid metabolite (FGM) levels between the two subherds. Fecal glucocorticoid metabolite levels varied by season in both subherds, with levels in winter lower than during the other three seasons. Lamb:ewe ratios were not different between the control and treatment subherds at the end of summer 1999. In contrast, the treatment group had a lower lamb:ewe ratio at the end of summer 2000 despite having lower LPG. However, this result was attributed to lower ewe production, not lower lamb survival. The LPG levels were not correlated with FGM concentrations; instead, FGM levels might reflect normal seasonal patterns. Other factors, including contagious ecthyma, were more important for determining lamb mortality than LPG and FGM levels during our study. We suggest further experimental work over a longer duration to address these relationships.

Animals↗
PubMed1989

Hepatitis B surface antigen (HBsAg) in feces of convalescent hepatitis B patients.

In 1983, 13 subjects were selected from a factory where an outbreak of viral hepatitis B occurred in October 1982. About 5-7 months later, 13 patients with viral hepatitis B were studied by RPHA and ELISA for HBsAg in feces. Presence of HBsAg was confirmed by specific blocking procedure. HBsAg was found in feces in 69% (9/13) of patients and 30.2% (26/86) of samples collected. 14 fecal specimens found to be positive for HBsAg by RPHA or ELISA examined by immune electron microscopy. Small aggregates of 20-28 nm HBsAg spherical particles were found in three fecal samples. 40-50 nm Dane's-like particles were also observed in one sample. The results suggested that in convalescent period of viral hepatitis B the HBsAg might be excreted through feces.

Convalescence↗
PubMed1987

[Multicausality in the pathology of commercial swine breeding].

Studied were more than 100,000 samples of pathologic tissue material, 1,900 whole blood and blood serum samples, 8,000 fecal samples, 2,600 washings samples, swab samples, and other sampled material via the employment of a number of bacteriologic, virologic, morphologic, hematologic, biochemical, parasitologic, and other routinely applicable laboratory methods over a period of 5 years. All materials were taken from 15 stock swine breeding complexes, 3 reproduction and 2 nucleus bases for hybrid pigs, and 3 swine breeding farms with a practice of extensive raising technology. It was demonstrated that the adaptive capacity of the animals of the Camborough hybrid sharply dropped when the pigs were kept under unconventional conditions of raising, leading to deviations in the application of industrial technology methods. This was the reason to observe the simultaneous manifestation of conditionally substantiated disease outbreaks (multiple causality), with great economic losses. Studied were the etiology and pathogenesis of coli bacteriosis, salmonellosis, gastric ulcer, hypotrophy, atrophic rhinitis, geohelminthiases, chlamydial infections, and erysipelas. Suggested is the use of programmes and methods of prophylaxis and treatment, of expedient diagnosis and control on immunity.

Animal Husbandry↗
PubMedDate not supplied

Clostridium difficile in a department of surgery.

The colonization and transmission of Clostridium difficile strains was investigated at the Department of Surgery. Anaerobes were isolated from fecal samples of 27 patients among 308 investigated (mean age: 59.5 years). The majority of them (22 out of 27 patients) received anti-microbial agents prior to investigation for Clostridium difficile. Eighteen of them had undergone various surgical procedures including laparotomies. Only four Clostridium difficile positive patients had diarrhea, but pseudomembranes were not observed endoscopically. The highest colonization rate of C. difficile (20/27) was found to occur in autumn. All 54 samples from medical personnel were C. difficile free. C. difficile was isolated in 7% of samples taken from hospital environment. Toxigenicity of Clostridium difficile strains isolated from fecal samples taken from patients and from environment was 78% and 80%, respectively, as determined by cytotoxicity tissue assay, or 70% and 20%, respectively, in the latex agglutination test.

Adult↗