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PubMed1991

Immunoglobulin E in the feces of children and adolescents from some tropical and subtropical countries.

Eighteen of 27 individuals, aged from 6 months to 19 years (mean 5 years, 7 months), from countries in the tropics or the subtropics had either intestinal parasitic infestations or intestinal enteropathogenic bacterial infections or both. Fourteen of those with intestinal pathogens had detectable concentrations of IgE in their fecal extracts, ranging from less than 0.5 to 420 IU/ml extract (mean 33 IU/ml). This rate of occurrence was significantly higher than the number of IgE-positive fecal extracts in a group of 54 healthy nonallergic Norwegian children (p less than 0.001), but did not differ from that of a group of 40 allergic children (p greater than 0.20). The individuals with intestinal helminthic infection had the highest fecal IgE concentrations. Of the 9 individuals who did not have any demonstrable intestinal pathogen, low concentrations of IgE could be detected in feces from only 2, which did not differ from the rate in the healthy Norwegian controls. The concentrations of IgE in the feces of the subjects from tropical/subtropical regions correlated linearly with the corresponding serum concentrations of IgE (r = 0.69; p less than 0.001). The results indicate that the combined load of intestinal pathogens, including helminths, protozoa, and enteropathogenic bacteria, may stimulate IgE production in the gut.

Adolescent↗

PubMed2000

Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.

Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.

Animals↗

PubMed2003

Comparison of two DNA extractions and nested PCR, real-time PCR, a new commercial PCR assay, and bacterial culture for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces.

In this study, 5 combinations of 2 DNA extractions and 3 polymerase chain reaction (PCR) techniques were compared with culture for the detection of Mycobacterium paratuberculosis directly from bovine feces. These combinations included a new commercial extraction technique combined with a commercial PCR/Southern blot technique, nested PCR (nPCR), or real-time PCR, and a university-developed extraction combined with nPCR or real-time PCR. Four of the 5 combinations had statistically similar sensitivities between 93% and 100% and specificity between 95% and 100%, when compared with culture results from 63 bovine fecal samples. These results indicated that using a commercial extraction with a commercial PCR/Southern blot, nPCR, or real-time PCR, or a university-developed extraction with real-time PCR would result in similar sensitivities to culture for the identification of M. paratuberculosis from bovine feces and are valid alternatives to culture.

Animals↗

PubMed2006

Use of pooled samples for the detection of Salmonella in feces by polymerase chain reaction.

Many epidemiological studies of Salmonella rely on conventional bacteriological culture methods to detect Salmonella in fecal samples. These culture-based methods are inefficient for epidemiological studies in populations with a low prevalence of Salmonella. The objective of this study was to optimize a protocol that uses pooled Salmonella enrichment broth cultures of bovine feces and polymerase chain reaction (PCR) for the detection of the invA gene of Salmonella in feces. In one field trial, 196 animals were sampled, and all samples were tested by culture, invA PCR on individual samples, invA PCR on pools of 5 samples, and BAX PCR on individual samples. All assays showed a high agreement on individual samples (kappa > or = 0.75). The invA PCR was run on each of 40 pools and detected 19 of 22 culture-positive pools. In another field trial, 152 samples were taken from 4 dairies, and the invA PCR was performed on pools of 5 samples in addition to bacteriological culture of individual samples. Salmonella was detected in 5 of the 32 pools (7 total positive samples) by both PCR and culture. One pool was PCR-positive but culture-negative. Pooling did not dramatically affect the performance of the invA PCR; most of the culture-positive samples were detected, including all of the samples when there were 4 or more Salmonella colonies on the agar plate. Based on these field trials, invA PCR on pooled samples appears to be an efficient method of Salmonella detection as long as Salmonella loads are not extremely low.

Animals↗

PubMed1993

Application of polymerase chain reaction for the correlation of Salmonella serovars recovered from greyhound feces with their diet.

The polymerase chain reaction was employed to correlate Salmonella serovars isolated from fecal material of greyhounds suffering from gastroenteritis with those isolated from the diet fed to the greyhounds prior to onset of diarrhea. Kennels around the Abilene, Kansas, area were contacted and supplied with materials needed to collect a portion of the diet each day. With the onset of diarrhea, the kennels were instructed to ship the fecal material and diet from the previous 10 days to the laboratory for testing. Forty-one fecal samples and corresponding diets were screened for Salmonella, Clostridium perfringens, Campylobacter jejuni, Staphylococcus aureus, Staphylococcus intermedius, and pathogenic (piliated) Escherichia coli by direct culture using standard procedures. The fecal material was also screened for coronavirus and parvovirus using electron microscopy. Thirty-five "normal" fecal samples were screened for all of the above mentioned microorganisms as a control. In addition, the fecal material was screened for E. coli verotoxins I and II and clostridial enterotoxins. A total of 61 Salmonella isolates were recovered from the 41 samples of feces and diet submitted for testing; 31 were recovered from the feces and 30 from the diet. Four Salmonella isolates were recovered from the normal fecal samples. Results obtained by PCR, plasmid profiles, antigenic analysis, and antibiogram profiles indicated that 16 of the 31 isolates recovered from the fecal material were the same strain as that recovered from the diet.

Animal Feed↗

PubMed2001

Simultaneous determination of glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid mono-glucuronide in Shakuyaku-kanzo-to incubated with rat feces by semi-micro high-performance liquid chromatography.

A method for semi-micro high-performance liquid chromatography (HPLC) has been established for the simultaneous determination of glycyrrhizin (GL), glycyrrhetic acid (GA) and glycyrrhetic acid mono-glucuronide (GAMG) in incubation mixtures of rat feces with Shakuyaku-kanzo-to decoction (combination of licorice root and peony root). The analysis could be accomplished within 20 min with a TSKgel ODS-80TsQA (150 x 2.0 mm i.d.) column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile at a flow rate of 0.2 ml x min(-1), a thermostatic oven at 25 degrees C, and detection at 254 nm. The detection limits of these compounds were 0.1-0.85 pmol per injection (5 microl). The concentrations of GL and its metabolites in the incubation mixture after continuous consumption of Shakuyaku-kanzo-to were significantly different compared with those of untreated control. GL-hydrolysis of rat feces was enhanced by pre-consumption of Shakuyaku-kanzo-to.

Anaerobiosis↗

PubMed2002

Development of a simple HPLC method for determination of paeoniflorin-metabolizing activity of intestinal bacteria in rat feces.

A simple and reproducible HPLC method for the determination of paeoniflorin (PF)-metabolizing activity of intestinal bacteria in rat feces was developed and validated. Orally administered PF, a major active constituent of Paeoniae Radix, is metabolized into a bioactive compound, paeonimetabolin I (PM-I) by intestinal bacteria. Direct determination of the PF-metabolizing rate into PM-I is hard to achieve by HPLC due to the lack of intense chromophore in PM-I. However, when PF was incubated with Lactobacillus brevis, an intestinal bacterium, in the presence of phenylmercaptan, the metabolizing rate of PF into 8-phenylthio-paeonimetabolin I (PT-PM-I) was found to be equivalent to that of PF into PM-I. Thus, the PF-metabolizing activity of intestinal bacteria in rat feces was determined by measuring the rate of biotransformation of PF into PT-PM-I, which was detected by HPLC at 255 nm. This method can be utilized in the biopharmaceutical study of traditional Chinese formulations containing Paeoniae Radix.

Animals↗

PubMed1993

Application of a DNA-DNA hybridization method for detection of Campylobacter jejuni in chicken feces.

A direct colony hybridization method was used for the detection of Campylobacter jejuni in chicken feces. The biotin-labeled DNA prepared from the whole genome DNA of C. jejuni subsp. jejuni ATCC 33560 reacted well with homologous DNA and slightly with C. coli DNA. The method with the probe was found to be sensitive enough to detect a small number (10(2) CFU/g) of C. jejuni in chicken feces which contained a large number of background flora. It was suggested that this simple and sensitive method was useful for a wide survey of C. jejuni.

Animals↗

PubMed2003

PCR detection of Porcine circovirus type 2 DNA in whole blood, serum, oropharyngeal swab, nasal swab, and feces from experimentally infected pigs and field cases.

Porcine circovirus type 2 (PCV2) shedding patterns were investigated by polymerase chain reaction (PCR) for the detection of PCV2 DNA, and the diagnostic suitability of a sample for the PCR was examined by using different types of samples. In the experimental infection, sixteen pigs were inoculated intranasally with PCV2. The samples, including oropharyngeal and nasal swabs, feces, whole blood and serum became positive for PCV2 DNA by PCR immediately after the inoculation, and almost all samples remained positive during the observation period, post-inoculation-day 70. Field samples were collected from 313 pigs in five different age groups. The overall percentages of positive samples in the whole blood, nasal swabs, and feces detected by PCR were 30.4%, 19.2%, and 20.4%, respectively. The frequency of positive samples increased after the nursery stages and reached a peak in the 3 to 4-month-old pigs. These results indicate that PCV2 infection may occur after weaning, that PCV2 DNA may be present in whole blood for a long period after infection, and that whole blood and serum are the most suitable sample types for the PCR analysis of PCV2.

Aging↗

PubMed2005

Bacteriological survey of feces from feral pigeons in Japan.

Some public areas in Japan such as parks and gardens can be highly contaminated with pigeon feces. We examined levels of four bacterial contaminations in fecal samples from feral pigeons in 7 prefectures. We isolated Salmonella Typhimurium and S. Cerro from 17 (3.9%) of 436 samples, as well as Mycobacterium spp. including M. avium-intracellulare complex from 29 (19.0%) of 153 samples. The polymerase chain reaction detected Chlamydia psittaci and C. pecorum in 106 (22.9%) of 463 samples, but E. coli O-157 was not isolated from any of the samples. Our results indicate that pigeon feces are a source of several zoonotic agents for birds, animals and humans.

Animals↗

PubMed2002

Detection by PCR of the Tyzzer's disease organism (Clostridium piliforme) in feces.

We examined whether the Tyzzer's disease organism, Clostridium piliforme, could be detected in feces by PCR. If the organism could be detected in feces, a diagnosis could be made without sacrifice of the animal. Using the RT strain of C. piliforme, we found that a C. piliforme band could be detected when there were > or = 1 x 10(0) bacteria present in the PCR solution, but the presence of fecal extract in the solution depressed the sensitivity 10 fold. Nevertheless, we could detect the C. piliforme-specific band in fecal extracts from rats in a naturally infected colony, and concluded that the use of PCR to detect C. piliforme DNA in fecal extracts would be a useful diagnostic technique.

Animals↗

PubMed1981

Bacterial flora of ex-germfree mice after oral inoculation of feces from various species of conventional animals.

Emulsions of fresh feces from a conventional mouse, rabbit, dog or cynomolgus monkey were administered orally to germfree and conventional ICR mice. Various fecal floras bearing a striking resemblance in pattern to those in the inoculant became established in the inoculated ex-germfree mice. Reinoculation of these ex-germfree mice with fecal emulsions from a conventional mouse made 21 days after the original inoculation (conventionalization) led to alteration of their floras to the indigenous pattern of conventional mice. The administration of feces from the conventional animals of different species did not appreciably modify the fecal flora of conventional mice. The results indicate that intestinal flora of animals is determined in the first stage by that of the contaminants, but this would be altered later under the predominant influence of the various host factors.

Animals↗

PubMed1993

[Relationship between the pathogenicity of Schistosoma mansoni in mice and the susceptibility of the mollusk vector. II. Number of eggs in feces and number and size of granuloma in visceras].

The influence of the susceptibility of the vector snails of S. mansoni on the development of the pathogenicity of the worm to the host was studied. Off-spring of snails Biomphalaria glabrata and Biomphalaria tenagophila were used, selected with regard to the susceptibility to S. mansoni of the strains BH and SJ trait, respectively. Parenteral, F1, F2, F3 and F4 generation cercariae were used for the infection of Swiss mice, which were killed eight weeks after infection. The number of eggs in the feces and the number of granulomas in the liver, spleen and intestines were counted. The size of the granulomatous reactions was evaluated. The results led to the conclusion that greater susceptibility of B. tenagophila was associated with a larger egg production in the feces and a higher density of granulomas in the liver. There is a clear inverse relationship between the area of hepatic, splenic and intestinal granulomas and the rate of infection of the snails which served as intermediate hosts for the S. mansoni.

Animals↗

PubMed1998

[Determination of HIV protein 24 in feces of Triatoma infestans fed with blood from AIDS patients].

OBJECTIVE: To verify any possibility of the elimination of p24 in feces of Triatoma infestans, p24 being an antigen and one of the markers of HIV infection. The purpose of the study was the evaluation of any risk of contamination by laboratory staff who work with activities related to this particular insect, and also investigate any mechanism that might thus lead to the dissemination of HIV. METHOD: Triatoma infestans were fed with p24-marked blood from 23 AIDS patients. Twenty-four and 48 hours later, their feces were examined in order to verify the presence of the antigen. The search for p24 was performed by means of the immunoenzymatic technique. RESULT AND CONCLUSION: No protein 24 was detected in any of the samples. In accordance with the applied methodology, the purpose of showing that the elimination of p24 does not occur, was obtained. Were this question approached in other ways different results might reveal new information in terms of risks of HIV dissemination.

Acquired Immunodeficiency Syndrome↗

PubMed2003

Terrestrial mammal feces: a morphometric summary and description.

The study of feces of terrestrial mammals brings out biological and ecological data such as the species presence, diet, behaviour, territory, parasitic fauna, and home-range use, which can be applied for conservation projects and support paleoecological research that use coprolites as the main source of study. Although the new biotechnological techniques allow more accurate data, the diagnosis based on morphometric analyses permits the primary identification of the taxonomic group origin to support the best choice of subsequent analyses. We present the compilation list of fecal shape and measurements available in the literature published in North America, Eastern and Southern Africa, Europe, and new data from Brazil. Shape and diameters are the best characteristics for taxonomic identification. Feces were assembled in 9 groups that reflect the Order, sometimes the Family, and even their common origin.

Africa↗

PubMed2005

Molecular characterization of trichomonads from feces of dogs with diarrhea.

Trichomonads are occasionally observed in the feces of dogs with diarrhea. On the basis of superficial morphological appearance, these infections have been attributed to opportunistic overgrowth of the commensal, Pentatrichomonas hominis. However, molecular characterization of canine trichomonads has never been reported. This study was performed to determine, by means of rRNA gene sequence analysis, the identity of trichomonads observed in feces from dogs with diarrhea. Total DNA was isolated from fecal samples obtained from a 3-mo-old mixed breed dog and litter of German Shepherd puppies having profuse liquid diarrhea containing numerous trichomonads. Total DNA was subject to PCR amplification of partial 18S rRNA gene or 5.8S, ITS1, ITS2, and partial 18S and 28S rRNA genes using species-specific and universal primers, respectively. Products of 642 and 1864 base-pair length were amplified and cloned. On the basis of rRNA gene sequence, the trichomonads observed in the single dog and the litter of puppies shared 100% identity with Tritrichomonas foetus and P. hominis, respectively. The present study is the first to establish the molecular identity of trichomonads infecting dogs with diarrhea. These studies validate the longstanding assumption that canine trichomoniasis may be attributed to P. hominis. Importantly, these studies additionally recognize that canine trichomoniasis may also be caused by infection with T. foetus.

Animals↗

PubMedDate not supplied

Phosphorus characteristics of dairy feces affected by diets.

Phosphorus (P) surplus on dairy farms, especially confined operations, contributes to P buildup in soils with increased potential for P loss to waters. One approach to reduce P surplus and improve water quality is to optimize P feeding and improve P balance on farms. Here we report how varying P concentrations in lactating cow diets affects the amount as well as the chemical forms and fraction distribution of P in fecal excretion, and the environmental implications of this effect. Analysis of fecal samples collected from three independent feeding trials indicates that increasing dietary P levels through the use of P minerals not only led to a higher concentration of acid digest total phosphorus (TP) in feces, but more importantly increased the amount and proportion of P that is water soluble and thus most susceptible to loss in the environment. For instance, with diets containing 3.4, 5.1, or 6.7 g P kg(-1) feed dry matter (DM), the water-soluble fraction of fecal P was 2.91, 7.13, and 10.46 g kg(-1) fecal DM, respectively, accounting for 56, 77, and 83% of acid digest TP. The other fecal P fractions (those soluble in dilute alkaline and acid extractants) remained small and were unaffected by dietary P concentration. Excess P in the P supplemented diets was excreted in feces as water-soluble forms. A simple measure of inorganic phosphorus (Pi) in a single water extract is highly responsive to changes in diet P concentrations and hence can be indicative of dietary P status. A fecal P indicator concept is proposed and discussed.

Animal Feed↗

PubMed2000

Improved culture methods for isolation of Salmonella organisms from swine feces.

OBJECTIVE: To compare 3 alternative culture techniques for the detection of Salmonella organisms in swine feces with a modification of the International Standard Organization (ISO) 6579 standard protocol. SAMPLE POPULATION: Fecal samples from swine herds suspected of having Salmonella infections. PROCEDURE: 4 experiments were performed to evaluate the following: 1) diagnostic sensitivity of the selective preenrichment and rapid isolation novel technology (SPRINT) protocol, compared with that of the modified ISO protocol; 2) detection limit of the SPRINT protocol for Salmonella organisms; 3) use of tetrathionate-novobiocin (TTN) broth, compared with selenite cysteine (SC) broth for selective enrichment; and 4) use of universal preenrichment (UPE) broth, compared with buffered peptone water (BPW) for preenrichment of samples prior to the use of modified semisolid Rappaport-Vassiliadis (MSRV) plates. RESULTS: Comparing the Salmonella culture results of 183 swine fecal samples, the diagnostic sensitivity of the SPRINT protocol (0.86) was not significantly different than the diagnostic sensitivity of the modified ISO protocol (0.80), although it was 24 hours faster. The SPRINT protocol could detect 5 of the 6 investigated Salmonella serotypes at inoculation concentrations of < 10 colony-forming units (CFU)/25 g of uncontaminated feces. The TTN broth performed significantly better than the SC broth for selective enrichment of Salmonella organisms. There was no significant difference in results of preenrichment of samples between the use of UPE broth or BPW. CONCLUSIONS AND CLINICAL RELEVANCE: The SPRINT protocol may provide a faster alternative for isolation of Salmonella organisms from swine fecal samples. Furthermore, the use of TTN broth instead of SC broth may increase the sensitivity of the modified ISO 6579 protocol.

Animals↗