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PubMed2003

Real-time PCR for the detection of Giardia lamblia.

Microscopy is considered to be the gold standard for diagnosis of Giardia lamblia infection. However, this method is time-consuming and not very sensitive. We developed a real-time PCR assay based on the small subunit ribosomal RNA gene of G. lamblia for the specific detection of G. lamblia DNA in stool samples and thereafter compared the results with microscopy and antigen detection. The G. lamblia real-time PCR was positive in 102 of 104 fecal samples known to contain G. lamblia cysts and was positive in 10 fecal samples in which G. lamblia antigen was detected but in which no cysts were found with microscopic examination of concentrated fecal samples. The real-time PCR is as specific and sensitive as antigen detection and is more sensitive than microscopy. Moreover, in two patients we were able to detect G. lamblia earlier in the course of infection than with any of the other methods.

Animals↗
PubMed2005

Clostridium difficile in emergency room.

Clostridium difficile strains are known as etiological agents of pseudomembranous colitis (PMC), antibiotic-associated diarrhea (AAC) and colitis (AAC) and hospital-acquired infections. The aim of this study was to determine the frequency of C. difficile infection among patients in the emergency room and to compare isolated strains by phenotypic and genotypic characteristics. During a period of 11 months, 56 stool samples taken from diarrheic patients hospitalized in the emergency room of the Medical Center UC Davis and 14 environmental samples were cultured for isolation of C. difficile strains. Eighteen C. difficile strains were isolated from stool samples cultured on selective TCCCA plates and 5 strains from environmental samples using Rodac plates. Eleven toxigenic (TcdA+/TcdB+), 6 non-toxigenic (TcdA-/TcdB-) and unique toxin A-negative/toxin B-positive (TcdA-/TcdB+) C. difficile strains were detected among patients' isolates and 3 toxigenic and 2 non-toxigenic strains-among environmental samples. The majority of C. difficile-positive patients were treated previously by antibiotics. Four strains isolated from patients' fecal samples and one strain isolated from the environment demonstrated high-level resistance to erythromycin and clindamycin (MIC >256mug/mL). The results obtained by AP-PCR and PCR-ribotyping revealed genetic heterogeneity among the strains isolated from patients' fecal samples. However, similarity was observed among environmental strains and strains isolated from patients' fecal samples. Considering the importance of emergency room patients as a potential source of C. difficile strains, it appears to be important examine these patients for C. difficile before transfer to the other hospital units.

Journal Article↗
PubMed2005

The absence of Rhodococcus equi in Mongolian horses.

In native Mongolian horses, the incidence and distribution of Rhodococcus equi are poorly understood. One hundred and fourteen equine fecal samples and 71 soil samples were collected from the camp sites of 26 nomadic families located in three areas less than 100 km from Ulaanbaatar, Mongolia. Five fecal samples were also collected from foals of Przewalski's Horses introduced into the Hustai National Park, Mongolia. No R. equi was isolated from the Mongolian horses or the soil samples. However, three colonies of R. equi were isolated from two fecal samples collected from foals of Przewalski's Horses. These isolates were avirulent, with neither 15- to 17-kDa antigens (VapA) nor a 20-kDa antigen (VapB) genes being detected. We concluded that native Mongolian horses and their environment appear free from contamination with R. equi.

Animals↗
PubMed1992

Serotype 2 reoviruses from the feces of cats with and without diarrhea.

During a virus survey carried out in the period 1989-90 with 148 fecal samples collected from cats in Japan, three reovirus strains were isolated in feline cell cultures. Two strains (Nos. 114 and 140) were from 48 diarrheal fecal samples and another strain (No. 32/41) was from 100 normal fecal samples. The strains grew in feline and simian cell cultures with producing typical intracytoplasmic inclusion bodies in which virus particles were densely packed. All strains, especially Nos. 32/41 and 140 strains, showed trypsin-dependent growth in vitro. Their ultrastructural and genomic properties were characteristic of genus reovirus in the Reoviridae. All strains agglutinated erythrocytes of human type O but not of bovine. Although they were identified as serotype 2 by hemagglutination-inhibition test with the hyperimmune sera against human reovirus prototype strains, No. 114 strain was typical and the other two strains were atypical serotype 2 reoviruses. Furthermore, from the reason that Nos. 32/41 and 140 strains possessed some common properties though derived from cats in distant locations, they were considered to be reoviruses having been maintained in the cat population. Seroepizootiologic survey revealed that the prevalence of serotype 3 infection was most widespread and serotype 2 was least among three serotypes of reovirus in a cat population.

Animals↗
PubMed2001

Selection of a potential probiotic Lactobacillus strain and subsequent in vivo studies.

The probiotic potential of a Lactobacillus strain, isolated from pig faeces, was assessed as a probiotic in piglets. The strain was examined for resistance to pH 2.0, 0.5% oxgall and antibiotics, and antimicrobial activities against enteric pathogenic bacteria. The probiotic strain, L. reuteri BSA131, was administered through the feed to 25 1-month-old Landrace piglets. The piglets were divided into five groups of five piglets each and fed with different diets for 28 days. The daily consumption of L. reuteri BSA131 was assigned into two groups by the concentration of 10(6) or 10(8) freeze-dried bacteria. Fecal samples were collected before, during, and after consumption. Lactobacilli and enterobacteria cell counts were determined in the fecal samples. The liveweight gains and feed consumption of the piglets were recorded daily. This study showed that strain BSA131 enhanced liveweight gains and feed conversion rates in piglets. It also showed a significant increase in lactobacilli cell counts and decreases in enterobacterial numbers in the fecal samples. Strain BSA131 was considered to be a potential probiotic for piglets, especially after weaning.

Animal Feed↗
PubMed1999

Factors associated with fecal shedding of verotoxin-producing Escherichia coli O157 on dairy farms.

Fecal samples were collected from 4,361 dairy cows on 91 dairy operations between 26 February and 8 July 1996. Fecal samples were cultured for Escherichia coli O157, and positive isolates were probed for verotoxin-producing genes. A total of 52 (1.2%) fecal samples on 22 (24.2%) operations were positive for verotoxin-producing E. coli O157. Herds in which samples were collected on or after 1 May 1996 were significantly more likely to test positive than herds sampled before that date (odds ratio = 7.7). Herds maintained on farms on which alleyways were flushed with water to remove manure were 8.0 times more likely to have samples test positive for verotoxin-producing E. coli O157 than were herds maintained on farms cleaned by use of other methods of manure removal.

Animal Feed↗
PubMed2006

[Viraemia and extraintestinal involvement after rotavirus infection].

OBJECTIVE: To study the incidence of viraemia and extraintestinal organ damage in children with acute rotavirus (RV) gastroenteritis. METHODS: Eighty-three children with acute rotavirus gastroenteritis were hospitalized from October 2002 to March 2003, whose blood and fecal samples were obtained on admission. Rotavirus RNA (encoding the VP7 outer capsid protein) were detected in blood and fecal samples by nest reverse transcription-polymerase chain reaction (RT-PCR). According to the result of blood RV-RNA, the patients were divided into RV-RNA positive group and RV-RNA negative group. The differences between these two groups in the severity of gastroenteritis and extraintestinal organ damage were analyzed. RESULTS: Eighty-two of 83 stool samples from the children with rotavirus infection were positive for rotavirus RNA. Sixteen of 83 blood samples were positive for rotavirus RNA with a positive rate of 19.3%. The nucleotide sequence of cloned cDNAs, resembling part of the VP7 gene, was identical from paired blood and fecal samples. There were no significant differences between blood RV-RNA positive group and blood RV-RNA negative group in the rate and degree of fever, diarrhea, dehydration, metabolic acidosis, hypokalemia and myocardial damage (P>0.05); while the incidences of liver damage, rash, lower respiratory tract infection and the central nervous system involvement in the blood RV-RNA positive group were significantly higher than those in the blood RV-RNA negative group (P<0.05). CONCLUSION: Viraemia is present in the children with acute rotavirus gastroenteritis. Viraemia might be an important mechanism by which rotavirus spread to the extraintestinal sites resulting in organs damage.

Base Sequence↗
PubMed2006

Prevalence of shiga toxin-encoding bacteria and shiga toxin-producing Escherichia coli isolates from dairy farms and county fairs.

Shiga toxin-encoding bacteria (STB) and shiga toxin-producing Escherichia coli (STEC) were detected and isolated from dairy cattle and their farm environment and from manure piles at Minnesota (MN) county fairs from 2001 to 2002. A total of 2,540 samples were collected from 28 dairy cattle farms (8 organic and 20 conventional), 17 calf pens (5 organic and 12 conventional), and 12 county fairs. STB were detected from 71 (3.2%) of 2208 fecal samples with 20 (71.4%) of 28 dairy farms having at least one positive animal sample. In samples collected from conventional farms, 41 (2.3%) of 1750 fecal samples were STB-positive and 13 (65%) of 20 farms had at least one positive animal. Thirty (6.6%) of 458 fecal samples from organic farms were STB-positive and 7 (87.5%) of 8 farms had at least one positive animal. STB was detected from 31 (17.4%) of 178 samples and 7 (58.3%) out of 12 manure piles at county fairs. A total of 43 STEC isolates were recovered and belonged to 26 different serotypes (19 O and 18 H types). Among STEC, 60.5% possessed only stx1, 30.2% stx2, and 9.3% both stx1 and stx2. The genes eae and hlyA were detected in more than 50% of the STEC isolates. STB can be found on most dairy cattle farms including organic and conventional herds and county fairs. The presence of these potentially pathogenic bacteria in county fairs may pose a risk to the public who have contact with cattle or their environment.

Animals↗
PubMed1994

Gastro-intestinal parasites of the Samango monkey, Cercopithecus mitis, in Natal, South Africa.

Eight gastro-intestinal tracts of Cercopithecus mitis labiatus from Karkloof, Natal, and 121 fecal samples from C. m. erythrarchus from Cape Vidal, Natal, were examined for helminth parasites and/or their eggs. Fecal samples from six of the C. m. labiatus were examined for protozoan cysts. Five protozoon and six helminth species were identified from C. m labiatus. Most adult worms occurred in the caecum and colon, gut regions which also contained the highest volatile fatty acid levels. The eggs of nine helminth species were recovered from C. m. erythrarchus fecal samples; protozoans were not looked for in these samples.

Animals↗
PubMed2006

Ribosomal RNA sequences of Clostridium piliforme isolated from rodent and rabbit: re-examining the phylogeny of the Tyzzer's disease agent and development of a diagnostic polymerase chain reaction assay.

We used polymerase chain reaction (PCR) technology to amplify the 16S rRNA gene, the intergenic spacer, and most of the 23S rRNA gene from 6 isolates (2 mice, 1 hamster, 1 rat, and 2 rabbit isolates) of the Tyzzer's disease agent (Clostridium piliforme) and C. colinum. Sequence similarity searches of GenBank identified 45 closely related bacteria, which we used for phylogenetic analysis by parsimony and maximum-likelihood methods using Escherichia coli to root the resulting phylogram. Microorganisms identified as C. piliforme form 3 clusters within a single clade; the nearest related distinguishable species is C. colinum. Other bacterial clades closely related to C. piliforme are clostridia previously identified by molecular methods in the bovine, porcine, and human gastrointestinal tracts. DNA sequence alignment highlighting sequence differences were used to design a rodent and rabbit C. piliforme-specific PCR assay, which targets a 639-basepair region at the 3' end of the 16S rRNA gene and the 5' end of the intergenic spacer. We used this PCR assay to examine 4 rat fecal samples from C. piliformeseropositive rats and reexamine 2 rabbit fecal samples previously identified as containing DNA sequences consistent with C. piliforme infection by 16S PCR assay. Our new assay did not detect the presence of C. piliforme DNA sequences in either the rat or rabbit fecal DNA samples, consistent with the absence of clinical disease in the colonies evaluated.

Animals↗
PubMed2004

Role of cell walls in the bioaccessibility of lipids in almond seeds.

BACKGROUND: Certain nutrients and phytochemicals in almonds may confer protection against cardiovascular disease, but little is known about factors that influence their bioavailability. A crucial and relevant aspect is the amount of these dietary components available for absorption in the intestine, which is a concept referred to as bioaccessibility. OBJECTIVE: We investigated the role played by cell walls in influencing the bioaccessibility of intracellular lipid from almond seeds. DESIGN: Quantitative analyses of nonstarch polysaccharides (NSPs) and phenolic compounds of cell walls were performed by gas-liquid chromatography and HPLC, respectively. In a series of experiments, the effects of mechanical disruption, chewing, and digestion on almond seed microstructure and intracellular lipid release were determined. In the digestibility study, fecal samples were collected from healthy subjects who had consumed diets with or without almonds. Almond seeds and fecal samples were examined by microscopy to identify cell walls and intracellular lipid. RESULTS: Cell walls were found to be rich in NSPs, particularly arabinose-rich polysaccharides, with a high concentration of phenolic compounds detected in the seed coat cell wall. During disruption of almond tissue by mechanical methods or chewing, only the first layer of cells at the fractured surface was ruptured and able to release lipid. In fecal samples collected from subjects consuming the almond diet, we observed intact cotyledonary cells, in which the cell walls encapsulated intracellular lipid. This lipid appeared susceptible to colonic fermentation once the cotyledonary cell walls were breached by bacterial degradation. CONCLUSION: The cell walls of almond seeds reduce lipid bioaccessibility by hindering the release of lipid available for digestion.

Adult↗
PubMed2003

Detection of porcine circovirus type 2 in feces of pigs with or without enteric disease by polymerase chain reaction.

To establish the sensitive polymerase chain reaction(PCR) method and detect porcine circovirus type 2 (PCV2) from intestines and feces of commercial swine herds with or without enteric disease, intestinal samples from 68 pigs and 29 fecal samples from commercial swine farms were collected. A primer set, forward primer 5'-GAAGAATGGAAGAAGCGG-3' and reverse primer 5'-CTCACAGCAGTAGACAGGT-3', could detect the virus at a concentration as low as 2 infectious virions per milliliter under controlled conditions using PK-15 cell-adapted PCV2. The genomic nucleotide sequences of open reading frame 1 (ORF1) PCR products from fecal samples were found to have complete homology with other PCV2s deposited in the GenBank database. Transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) as the other enteric pathogens were also investigated by performing duplex reverse transcription-PCR (RT-PCR). Among 63 pigs with clinical enteric disease, 18 PCV2s (14 from intestines and 4 from feces), 7 TGEVs from intestines, and 18 PEDVs (14 from intestines and 4 from feces) were detected by PCR and the duplex RT-PCR. In 34 pigs (14 from intestines and 20 from feces) without clinical enteric disease, only PCV2 was detected in 19 pigs (3 from intestines and 16 from feces). Both PEDV and PCV2 were found in 6 pigs with clinical enteric disease. Among 15 PCV2 samples that were PCR-positive, 4 were culture-positive at passage level 3 in PK-15 cells. These results reveal that PCV2 is shed through the feces of pigs without clinical enteric disease, which suggests the potentiality of the fecal-oral transmission of PCV2 in feces.

Animals↗
PubMed1994

Effects of diet on fecal occult blood testing in healthy dogs.

Six dogs were fed each of nine diets to evaluate the effects of diet on fecal occult blood test results. The diets represented a range of different type (i.e. canned, dry or semi-moist), protein and vegetable constituents, and fiber contents. Each diet was fed twice daily for five consecutive days; fecal samples were collected twice daily on days 4 and 5. An o-tolidine test kit and a guaiac paper test kit for fecal occult blood were used. Two hundred and sixteen fecal samples were analyzed (24 samples/diet). When using the guaiac test the following positive results were obtained from fecal samples from dogs consuming a canned meat- and vegetable-based diet (24/24 samples); a canned meat-based diet (24/24 samples); a dry corn and poultry-based diet (9/24 samples); a dry corn, wheat, and meat meal diet (4/24 samples), a canned poultry-based diet (1/24 sample) and a semi-moist soybean meal-based diet (2/24 samples). A total of 64 samples were positive using the guaiac test. Using the o-tolidine test, no samples were positive. The difference between the number of positive results with each test kit was highly significant (p < 0.001). Results indicate that 1) diet affects the specificity of guaiac test fecal occult blood results in the dog and 2) positive o-tolidine test results were not caused by diets fed in the study.

Animal Feed↗
PubMed2003

Escherichia coli O157 in feedlot cattle feces and water in four major feeder-cattle states in the USA.

The prevalence of Escherichia coli O157 was determined in 10662 fecal samples, 2130 water and 1132 water tank-sediment samples collected during the summer months in 2001 from 711 pens in 73 feedlots located in Kansas, Nebraska, Texas, or Oklahoma, USA. Overall, 10.2% of fecal samples were positive for E. coli O157, with 52% of the pens and 95.9% of the feedlots having at least one positive fecal sample. There were no differences among states or months in the fecal prevalences. Water or water tank-sediment was positive in 13.1% of the water tanks, and 60.3% of feedlots had at least one positive tank. Cattle were more likely to be shedding E. coli O157 in pens with positive water tanks, and water was more likely to be positive when E. coli O157 was detected in the sediment.

Animals↗
PubMed1987

Isolation of influenza A viruses from migratory waterfowls in San-in District, Western Japan in winters of 1980-1982.

In the two winters of 1980-1982, we surveyed migratory waterfowl of some species staying in San-in District, Western Japan for influenza virus at a few stations. From November 1980 to April 1981, only two strains of influenza virus, H13N1 and H11N6 subtypes, were isolated from 465 fecal samples from pintails but none from 255 samples from whistling swans nor from 625 black-tailed gulls. From November 1981 to March 1982, 17 viruses were isolated from 1156 fecal samples. Fourteen viruses, 10 H7N3, 2 H1N6 and 2 H3N8, were isolated from 459 feces samples from whistling swans. Two viruses, H13N3 and H13N6 subtypes, were isolated from 425 fecal samples from black-tailed gulls. A strain belonging to H1N3 subtype was isolated from 30 feces samples from mallards but no virus was isolated from 242 samples from pintails.

Animals↗
PubMed1995

Diagnosis of liver flukes in cows--a comparison of the findings in the liver, in the feces, and in the bile.

Percutaneous ultrasound-guided cholecystocentesis and aspiration of bile were attempted in 176 cows. These same procedures were performed in another 100 cows immediately after slaughter. The bile samples were examined microscopically for large and small liver fluke eggs. In addition, a fecal sample from each cow was examined for liver fluke eggs. The findings of both groups were summarized, and the results of the fecal and bile sample examinations were compared. In all cows the liver was examined for flukes, and the results were used as a reference. Of 41 cows in which adult flukes were found in the liver, 28 had F. hepatica eggs in fecal samples and 40 had F. hepatica eggs in bile samples. Of 204 cows in which no adult flukes were found in the liver, 23 had F. hepatica eggs in fecal samples and 27 had F. hepatica eggs in bile samples. The sensitivity of the determination of F. hepatica eggs in fecal and bile samples was 68 and 98%, respectively. The negative predictive values for fecal and bile examination were 93 and 99%, respectively. Of 49 cows in which adult flukes were observed in the liver, 13 had D. dendriticum eggs in fecal samples and 44 had D. dendriticum eggs in bile samples. Of 176 cows in which no adult flukes were found in the liver, 19 had D. dendriticum eggs in fecal samples and 49 had D. dendriticum eggs in bile samples. The sensitivity of the determination of D. dendriticum eggs in fecal and bile samples was 27 and 90%, respectively. The negative predictive values for fecal and bile examination were 81 and 96%, respectively. The results of this study indicate that the examination of bile is clearly a more reliable method of diagnosing liver fluke infections than microscopic examination of feces.

Animals↗
PubMed1995

Experimental oral inoculations in birds to evaluate potential definitive hosts of Neospora caninum.

Experimental oral inoculations to evaluate potential definitive hosts of Neospora caninum were conducted by feeding infected rodent tissues to 9 carnivorous birds of 4 species. Birds included 2 red-tailed hawks (Buteo jamaicensis), 2 turkey vultures (Cathartes aura), 2 barn owls (Tyto alba), and 3 American crows (Corvus brachyrhynchus). The rodents (mice or rats) had been inoculated with 100,000 culture-derived tachyzoites of N. caninum 1-6 mo before feeding to the birds. Fecal samples were collected from each bird daily for 1 mo after feeding rodents and examined for oocysts by fecal flotation. In addition, processed aliquots from all avian fecal samples were fed to BALB/c mice. Five weeks after feeding, mice were bled and sera were tested for antibodies against N. caninum. One to two months later, mice were killed and brain tissue was examined microscopically for protozoal cysts. While occasional oocysts were found in avian fecal samples, these were likely not N. caninum because they were not infective to BALB/c mice. It was concluded that the bird species tested are not likely to be definitive hosts of N. caninum.

Animals↗
PubMed1994

Steatocrit: a reliable semiquantitative method for detection of steatorrhea.

Steatocrit is a semiquantitative method for determination of fat content in fecal samples. Previous studies, mostly performed in children, reported controversial results. The aim of our study was to compare the determination of fat content in 148 fecal samples by two methods: the conventional van de Kamer and the steatocrit. Seventy-seven fecal samples had steatorrhea (> 7 g/day). The upper normal limit for the steatocrit (determined by the mean +/- 2 SD of samples without steatorrhea) was 2.1%. The steatocrit showed a sensitivity of 87%, specificity of 97%, and positive and negative predictive values of 97 and 87%, respectively. When fecal fat excretion > 20 g/day was evaluated, sensitivity increased to 98%. A significant linear correlation was found between steatocrit and the quantitative chemical method (r = 0.80; p < 0.0001). In conclusion, the steatocrit is satisfactory in the discrimination of patients with and without fat malabsorption. It is a simple, rapid, inexpensive, and reliable semiquantitative test that can be used when other methods are impractical.

Adult↗