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PubMed2005

Phosphatidylcholine and lysophosphatidylcholine excretion is increased in children with cystic fibrosis and is associated with plasma homocysteine, S-adenosylhomocysteine, and S-adenosylmethionine.

BACKGROUND: Hepatic steatosis and fat malabsorption are common in cystic fibrosis (CF). Choline deficiency results in decreased phosphatidylcholine synthesis through the cytidine diphosphocholine-choline pathway and hepatic steatosis and in increased synthesis of phosphatidylcholine from phosphatidylethanolamine using methyl groups from S-adenosylmethionine. The intestinal absorption of phosphatidylcholine in CF is unknown. OBJECTIVES: The objective was to determine whether excretion of choline phosphoglyceride (phosphatidylcholine and lysophosphatidylcholine) is increased in CF and whether loss of fecal choline phosphoglyceride is associated with altered plasma methionine cycle metabolites. DESIGN: A cross-sectional study involved 53 children with CF and 18 control children without CF. Blood was collected from all participants. A subset of 18 children with CF and 8 control children provided 72-h fecal samples and 5-d food records. RESULTS: Fat absorption was significantly lower (x+/- SEM: 86.2 +/- 1.6% and 94.1 +/- 1.2%) and excretion of fecal fat (12.9 +/- 1.7 and 3.9 +/- 0.7 g/d), phospholipid (median: 130 and 47.7 mg/d), phosphatidylcholine (19.6 and 2.1 mg/d), and lysophosphatidylcholine (60.3 and 16.9 mg/d) was significantly higher in children with CF than in control children, respectively (P < 0.05). Choline phosphoglyceride excretion was positively correlated with plasma homocysteine and S-adenosylhomocysteine and inversely related with plasma methionine (P < 0.05). CONCLUSIONS: Choline phosphoglyceride excretion is increased in children with CF and is associated with decreased plasma methionine and increased homocysteine and S-adenosylhomocysteine. These findings suggest choline depletion and an increased choline synthesis by S-adenosylmethionine-dependent methylation in CF, as well as a metabolic link between phosphatidylcholine metabolism and the methionine-homocysteine cycle in humans.

Case-Control Studies↗
PubMed1998

Non-invasive detection of fecal protein kinase C betaII and zeta messenger RNA: putative biomarkers for colon cancer.

We have developed a non-invasive method utilizing feces, containing sloughed colonocytes, as a sensitive technique for detecting diagnostic colonic biomarkers. In this study, we used the rat colon carcinogenesis model to determine if changes in fecal protein kinase C (PKC) expression have predictive value in monitoring the neoplastic process. Weanling rats were injected with saline or azoxymethane (AOM) and 36 weeks later fecal samples and mucosa were collected, poly A+ RNA isolated, and quantitative RT-PCR performed using primers to PKC betaII and zeta. Fecal PKC betaII and zeta mRNA levels were altered by the presence of a tumor, with tumor-bearing animals having a 3-fold higher (P < 0.05) PKC betaII expression as compared with animals without tumors. In addition, AOM-injection increased mucosal PKC betaII mRNA expression compared with saline controls. No effect of tumor incidence on mucosal PKC betaII expression was observed. In contrast, fecal PKC zeta expression was 2.5-fold lower (P < 0.05) in animals injected with azoxymethane versus saline. Since tumor incidence exerts a reciprocal effect on fecal PKC betaII and zeta mRNA expression, data were also expressed as the ratio between PKC betaII and zeta. The isozyme ratio was strongly related to tumor incidence, i.e. ratio for animals with tumors was 2.18 +/- 1.25, animals without tumors was 0.50 +/- 0.16, P = 0.025. We demonstrate that the expression of fecal PKC betaII and zeta may serve as a noninvasive marker for development of colon tumors. A sensitive technique for the detection of colon cancer is of importance since early diagnosis can substantially reduce mortality.

Animals↗
PubMed1978

Sources of Salmonellae in broiler chickens in Ontario.

Sources of Salmonellae infecting broiler chicken flocks in Ontario were investigated from July, 1975 to April, 1976. Three broiler flocks were investigated on each of four farms which received chicks from a common hatchery. Samples of feed and new litter were preenriched in nonselective broth subcultured to Salmonella-selective enrichment broth and plated on Salmonella-selective differential agar. Samples of used litter, water, culled chicks, insects, mice, wild birds and environmental swabs were not cultured initially in the nonselective broth. Fecal samples from principal and occasional flock attendants were examined for Samonellae. Salmonella infection, as judged by contaminated flock litter was detected in six flocks on two of the farms while the flocks on the other farms remained negative. Salmonellae were isolated from 23 of 412 feed samples (nine serotypes), six of 35 new wood shaving samples (four serotypes), one of 29 pools of culled chick viscera (one serotype) and 44 of 267 used litter samples (14 serotypes). These results indicate that broiler chicken flocks were infected with diverse Salmonellae introduced in day old chicks, pelleted feeds, wood shavings and residual contamination from the preceding flock.

Animal Feed↗
PubMed2001

Fecal analysis of ovarian cycles in female black-handed spider monkeys (Ateles geoffroyi).

An enzyme immunoassay (EIA) was applied to characterize the reproductive endocrinology of adult female black-handed spider monkeys (Ateles geoffroyi). Analysis of paired urine and fecal samples, collected from two females housed at San Diego Zoo, confirmed that the EIAs employed provided quantitative measurements of ovarian sex steroid hormones. Fecal metabolite levels were significantly correlated with those in urine, confirming that feces are a valid source of steroid metabolites in this species. The excretion of these metabolites in feces lagged urinary excretion by 1-2 days. The ovarian cycle profiles of the two captive females and five free-ranging females are comparable, with an average length of approximately 20-23 days. Cyclical bleeding, as previously reported, was observed in one of the two captive females. Pregnancy was detected in four free-ranging females, and early fetal loss for one female was indicated by hormonal data.

Animals↗
PubMed1993

Role of gastric pH in isolation of Helicobacter mustelae from the feces of ferrets.

BACKGROUND: Helicobacter mustelae colonizes the gastric mucosa of ferrets and causes persistent chronic gastritis. METHODS: Hypochlorhydria, as measured by gastric pH probe, was induced by administering oral omeprazole, a proton pump inhibitor of the parietal cell, to adult ferrets in two separate experiments. Feces of ferrets were cultured for H. mustelae before, during, and after omeprazole therapy. RESULTS: H. mustelae was isolated in 23 of 55 (41.8%) sequential fecal samples collected during omeprazole therapy. The same ferrets with acidic gastric pH had H. mustelae isolated in 6 of 62 (9.7%) of the fecal cultures (P < 0.01). A DNA species-specific H. mustelae probe confirmed the presence of the organism in both the stomach and feces of all 5 ferrets. In 4 of 5 ferrets restriction enzyme patterns of the gastric H. mustelae were identical to those of the fecal H. mustelae strains. CONCLUSIONS: Hypochlorhydria promotes fecal transmission of a gastric Helicobacter organism. The H. mustelae-colonized ferret provides an ideal model to study the epidemiology and pathogenesis of Helicobacter pylori-induced gastritis.

Animals↗
PubMed1985

Variation among hepatitis A virus strains. I. Genomic variation detected by T1 oligonucleotide mapping.

The genomes of eight hepatitis A virus (HAV) strains originating from far distant geographic regions such as Europe, North Africa, Middle and North America, Australia and The People's Republic of China were compared by RNase T1 oligonucleotide mapping. For this purpose, the viruses were propagated in cell cultures and viral RNA was isolated from highly purified mature virions. It could be shown that variation in nucleotide sequence is common among HAV isolates, but is in the order of magnitude reported for other picornaviruses. For viruses isolated in cell culture directly from stool samples of diseased individuals, changes usually amounted to 1-4% of RNA genome sites. Genomic differences between two virus strains derived from one fecal sample but replicating at either 32 or 37 degrees C were in the same order of magnitude. Thereby, the number of consecutive in vitro passages proved to have only limited influence on the development of genetic variation. For two HAV strains, however, adaptation to and passage in marmosets evidently had imposed highly selective conditions which had favored the appearance of viral genomes differing in up to 75% of their large oligonucleotides (about 10% in sequence) from the oligonucleotide map of a reference HAV strain.

Animals↗
PubMed1980

Fecal factors which modify the formation of fecal co-mutagens in high- and low-risk population for colon cancer.

Dietary patterns and fecal mutagens of 2 population groups with a distinct risk for colon cancer, high-risk, non-Seventh-Day Adventists and a low-risk, vegetarian Seventh-Day Adventists, in the New York metropolitan area were studied. The diet histories indicate that the intake of protein was identical in the 2 groups; a greater portion coming from meat in non-Seventh-Day Adventists and from vegetables in the Seventh-Day Adventists. The intake of fat was lower in Seventh-Day Adventists. Fecal samples were extracted with dichloromethane, partially purified on silica and assayed for mutagenic activity using the Salmonella/mammalian microsome mutagenicity test. When tested alone, fecal extracts from both groups were non mutagenic in TA98 and TA100 strains. Fecal extracts of non-Seventh-Day Adventists and Seventh-Day Adventists enhanced the mutagenic activity induced by 2-AAF in TA98 and TA100 (co-mutagenic activity). However, non-Seventh-Day Adventists showed a significantly higher co-mutagenic activity in TA98 than did Seventh-Day Adventists.

2-Acetylaminofluorene↗
PubMed2006

The endocrinology of pregnancy and fetal loss in wild baboons.

An impressive body of research has focused on the mechanisms by which the steroid estrogens (E), progestins (P), and glucocorticoids (GC) ensure successful pregnancy. With the advance of non-invasive techniques to measure steroids in urine and feces, steroid hormones are routinely monitored to detect pregnancy in wild mammalian species, but hormone data on fetal loss have been sparse. Here, we examine fecal steroid hormones from five groups of wild yellow baboons (Papio cynocephalus) in the Amboseli basin of Kenya to compare the hormones of successful pregnancies to those ending in fetal loss or stillbirth. Using a combination of longitudinal and cross-sectional data, we analyzed three steroid hormones (E, P, GC) and related metabolites from 5 years of fecal samples across 188 pregnancies. Our results document the course of steroid hormone concentrations across successful baboon pregnancy in the wild and demonstrate that fecal estrogens predicted impending fetal loss starting 2 months before the externally observed loss. By also considering an additional 450 pregnancies for which we did not have hormonal data, we determined that the probability for fetal loss for Amboseli baboons was 13.9%, and that fetal mortality occurred throughout gestation (91 losses occurred in 656 pregnancies; rates were the same for pregnancies with and without hormonal data). These results demonstrate that our longstanding method for early detection of pregnancies based on observation of external indicators closely matches hormonal identification of pregnancy in wild baboons.

Animals↗
PubMed1994

Sorrento studies of diet and fecal flora in the newborn.

The fecal flora of a breast-fed baby is very different from that of a bottle-fed baby. This paper reviews five previous studies, performed at this hospital concerning the effect of various dietary components (whey proteins, casein, lactoferrin, iron, nucleotides) on the fecal flora. The babies received either breast milk or one of the test formulas from birth. Fecal samples were examined by quantitative microbiological methods at 4 and 14 days and at various intervals thereafter. By 14 days differences in the fecal flora were established. Among breast-fed babies bifidobacteria, lactobacilli and staphylococci were predominant organisms, whereas in the formula-fed babies the predominant organisms were enterococci, coliforms, and Bacteroides. A whey-based formula without bovine lactoferrin, iron or nucleotides gave a flora a little closer to but still remote from the breast-fed one. Despite extensive modification of cow's milk in the manufacture of a modern infant formula, the fecal flora of bottle-fed babies remains substantially different from that of breast-fed babies.

Animals↗
PubMed2004

The stress response to environmental change in captive cheetahs (Acinonyx jubatus).

The captive North American cheetah (Acinonyx jubatus) population is not self-sustaining because of high prevalences of unusual diseases and poor reproductive success. Cheetahs are commonly moved between zoos for breeding purposes to maintain genetic diversity within the captive population, and movement may exacerbate infertility and disease. Fecal corticoids were analyzed by radioimmunoassay to measure the stress response of cheetahs to movement between facilities. Fecal samples were collected from 15 cheetahs for 14 days before movement and for at least 30 days after movement. For each cheetah, premovement fecal corticoid concentrations were used to determine baseline and then compared with trends in postmovement concentrations. In general, postmovement corticoid concentrations either increased (n = 8), did not change (n = 2), or decreased (n = 5). Although individual animal differences occurred, corticoid concentrations increased for most animals moved on-exhibit and decreased in animals moved off-exhibit. Animals moving on-exhibit had an 18-times greater risk of having corticoids elevated more than two standard deviations above baseline for 30 days after movement compared with animals that moved off-exhibit. In addition, greater day-to-day variation in corticoids occurred in animals moved on-exhibit. In general, animals with initially low baseline corticoid concentrations had a greater postmovement corticoid response than cheetahs with initially high baseline levels. These results indicate that some cheetahs have a prolonged stress response when moved between facilities, and the magnitude and character of this response is influenced by the exhibit environment.

Acinonyx↗
PubMed2000

A longitudinal study of Escherichia coli strains isolated from captive mammals, birds, and reptiles in Trinidad.

A longitudinal study was conducted of the prevalence and characteristics of Escherichia coli in mammals, birds, and reptiles housed at the Emperor Valley Zoo, Trinidad. During a 6-mo study period, swabs were obtained from fecal samples that were randomly collected from the enclosures of animals from these three taxonomic groups every 3 wk. With snakes, both cloacal and fecal swabs were obtained. Fecal and cloacal swabs were cultured for E. coli on eosin methylene blue agar. The production of mucoid colonies and hemolytic colonies and non-sorbitol fermenter status were identified. The occurrence of O157 strains was determined amongst E. coli isolates that were non-sorbitol fermenters, and the disc diffusion method was used to determine the antibiograms of isolates. The frequency of E. coli isolation was significantly higher in mammals compared with birds and reptiles. Overall, the frequencies of isolation of E. coli from omnivores. herbivores, and carnivores, 87.2%, 70.0%, and 57.3%, respectively, regardless of animal class, were significantly different. Most (99.6%) of the E. coli isolates tested for antibiotic sensitivity exhibited resistance to one or more of the eight antimicrobial agents used. The possession of phenotypic virulence markers by the E. coli isolates studied and the generally high resistance to antimicrobial agents may have health implications for the zoological collection.

Animals↗
PubMed1985

63Ni electron-capture gas chromatographic assay for buprenorphine and metabolites in human urine and feces.

A 63Ni electron-capture gas chromatographic assay is described for buprenorphine, a potent narcotic agonist--antagonist. In addition, the assay is useful for the measurement of the metabolite norbuprenorphine and demethoxybuprenorphine, a rearrangement product resulting when buprenorphine is exposed to acid and heat. An extraction procedure was developed which optimized recovery of buprenorphine from biological samples and produced minimal background interferences and emulsion problems. Extract residues were derivatized with pentafluoropropionic anhydride and assayed by gas chromatography. Samples were analyzed with and without enzyme hydrolysis, thus providing a selective and sensitive assay for both free and conjugated buprenorphine, norbuprenorphine and demethoxybuprenorphine. The lower limits of detection following extraction of a 1-ml sample were ca. 10 ng/ml for buprenorphine and demethoxybuprenorphine and 5 ng/ml for norbuprenorphine. Application of the assay to human samples following a 40-mg oral dose of buprenorphine produced no evidence for the presence of demethoxybuprenorphine in urine or feces. Norbuprenorphine (free and conjugated) was present in urinary and fecal samples; buprenorphine (free and conjugated) was found in high amounts only in feces and in trace amounts in urine as conjugated buprenorphine. The urinary and fecal excretion pattern observed for a human subject following oral dosing of buprenorphine suggests enterohepatic circulation of buprenorphine.

Adult↗
PubMed2006

The onset of puberty in North American bison (Bison bison) bulls.

Prepubertal testicular development was evaluated in young North American bison (Bison bison) bulls over a 12 months period, by following various parameters of semen quality intended to measure sexual maturity level. The study goals were to characterize bison male reproductive development and to define the age and onset of puberty. Semen was collected by electro-ejaculation from 12 bulls once a month for a period of 12 months, from 13 through 24 months of age. Volume, density, gross motility, individual motility, morphology, live/dead ratio, and concentration were used to judge the quality of the ejaculates. Abnormal sperm were classified into categories according to descriptions used for bovine evaluations and guidelines created by the Western Canadian Association of Bovine Practitioners were used for determining satisfactory semen quality. Minimums of 60% progressive motility and 70% normal sperm cells identified samples as satisfactory. Puberty was defined as the point in testicular development where an ejaculate contains a minimum of 50 x 10(6)sperm cells, showing at least 10% progressive motility. Fecal samples collected from bulls prior to electro-ejaculation for testosterone determination by a commercial testosterone specific antibody radioimmunoassay showed no significant changes in mean fecal testosterone concentrations over time. Changes in mean body weights over the study period were calculated, the average body weight at 13 and 24 months of age were 263+/-18.6 and 475+/-52.2 kg, respectively. Under these experimental conditions, age was determined to be the most significant factor in determining the onset of puberty. On average, bulls reached puberty at 16.5+/-2.5 months. There was a significant increase in the number of normal sperm with increasing bull age as well as significant decreases in proximal cytoplasmic droplets and head defects, but no significant changes in midpiece and principal piece defects. There were significant increases with age in individual progressive motility, gross motility, concentration, density, and volume but not in the percent of live sperm. Bison bulls used for breeding should be capable of passing a breeding soundness evaluation by 24 months of age. The commercial bison industry semen tests, and possibly rejects, many developmentally incomplete bulls between 19 and 21 months of age. We suggest that these may actually be developing normally and should not be hastily discarded; it may be worth testing again at 24 months.

Age Factors↗
PubMed2004

Equine Cryptosporidium parvum infections in western Poland.

A total of 564 fecal specimens from 318 horses used for recreational riding, child hippotherapy, and racing at ten commercial and government-run stables in western Poland were tested for Cryptosporidium parvum oocysts by microscopic examination of Ziehl-Neelsen stained smears, enzyme immunoassay, and combined direct immunofluorescent antibody and fluorescent in situ hybridization. Also, seven stool specimens from five personnel who had repeated contact with these horses were tested for C. parvum oocysts. Eleven horses that shed C. parvum oocysts were found in five of ten stables (50%). The prevalence of infection varied from 0% to 11.5%. The overall prevalence of equine C. parvum-associated cryptosporidiosis in the Wielkopolska region of western Poland was 3.5%. C. parvum oocysts were found only in fecal samples from mature horses, the number of oocysts was low, and infections were not associated with clinical signs. Oocysts were not found in human fecal specimens.

Animals↗
PubMed1995

Evaluation of a commercially available ELISA assay for detection of Giardia lamblia in fecal specimens.

A total of 417 fecal samples preserved in 10% buffered formalin and PVA were submitted to a commercial microbiology laboratory only for the detection of Giardia lamblia. Results from fecal specimens collected from 411 patients with gastrointestinal symptoms were compared using the following methods: (a) standard Ova & Parasite (O&P) concentration; (b) Alexon's ProspecT/Giardia enzyme-linked immunosorbent assay (ELISA) test, and (c) Meridian's Direct Fluorescent Antibody (DFA) Stain. In the 29 specimens in which G. lamblia was detected, 10 were O&P, DFA and ELISA positive, 17 were only ELISA positive and two were only Ova & Parasite and Direct Fluorescent Antibody positive. Of the 29 positive specimens, 22 were confirmed as true positives. The ELISA sensitivity was 91% and the specificity was 98%. The expense associated with these methods to detect the presence of Giardia is $11.00, $8.95, and $12.80, respectively. In symptomatic patients, the ProspecT/Giardia ELISA is a cost-effective, rapid, and sensitive method for detecting the presence of G. lamblia in fecal specimens.

Animals↗
PubMed2005

SARS coronavirus detection methods.

Using clinical samples from patients with severe acute respiratory syndrome, we showed that the sensitivities of a quantitative reverse transcription-polymerase chain reaction (80% for fecal samples and 25% for urine samples) were higher than those of the polyclonal (50% and 5%) and monoclonal (35% and 8%) antibody-based nucleocapsid antigen capture enzyme-linked immunosorbent assays.

Enzyme-Linked Immunosorbent Assay↗
PubMed2002

Distribution of Salmonella in swine production ecosystems.

The objective of this 2-year field survey was to sample multiple ecological compartments within swine production systems to identify potential sources of Salmonella infection for swine. Twelve single-site production systems within Illinois were identified by slaughter sampling to have detectable Salmonella in swine and therefore selected for study. There were four visits to each farm during a 5-month period. Fecal samples were obtained from swine and other wild and domestic mammals. Arthropods and environmental samples of feed, water, pen floors, boots, and bird feces were also collected. All 8,066 samples obtained were cultured to detect Salmonella. Salmonella was detected on 11 of the 12 farms. There were 206 positive cultures, including samples from swine (83), pen floors (54), boots (32), flies (16), mice (9), cats (3), and birds (3). Swine shedding Salmonella in feces were detected on 9 of the 12 farms. The more Salmonella-abundant ecological compartments were cats (12% of samples positive), boots (11%), bird feces (8%), flies (6%), and mice (5%); 2.1% of 4,024 swine samples were positive. All 221 feed samples were negative for Salmonella. There was a correlation between a farm having a high prevalence of shedding Salmonella in pigs and a high abundance on pen floors, flies, and boots. The most common serotypes detected were Derby, Agona, Worthington, and Uganda, which were distributed throughout the ecosystem, suggesting widespread transmission across ecological compartments. The ubiquitous distribution of Salmonella suggests that an effective control strategy must target multiple compartments of the swine production ecosystem.

Animal Husbandry↗
PubMed2005

A study of the prevalence and enumeration of Salmonella enterica in cattle and on carcasses during processing.

Salmonella prevalence and counts were estimated for samples from the oral cavity, hide, rumen, and feces of 100 cattle at slaughter and from the pre- and postchill carcasses of these cattle. Samples were collected from 25 consecutively slaughtered cattle from each of four unrelated groups slaughtered at a single abattoir on different days. Ten additional fecal samples from each group were collected from their respective abattoir holding pens prior to slaughter. The prevalence of Salmonella was estimated using automated immunomagnetic separation, and the counts were estimated using a combination of most probable number (MPN) and automated immunomagnetic separation. A total of 606 samples were collected with Salmonella isolated from 157 (26%), including 29% of oral cavities, 68% of hides, 16% of feces collected after evisceration, 25% of rumen samples, 2% of prechill carcasses, 3% of postchill carcasses, and 48% of feces collected from holding pens. The prevalence and count of Salmonella varied between the different groups of animals tested. The highest count obtained was from a rumen sample (1.1 x 10(4) MPN/g). Other counts were generally low, with a maximum count in feces collected after evisceration and in the abattoir holding pens of 93 and 23 MPN/g, respectively. The highest count on hides, in oral cavities, and on carcasses was 4.8 MPN/cm2, 23 MPN/g, and 0.31 MPN/cm2, respectively. Even though Salmonella was present on the hides and in the rumen and feces of at least one animal from each group of cattle, the processing of animals at this abattoir resulted in few contaminated carcasses, and when contamination occurred, Salmonella was detected at low numbers.

Abattoirs↗