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Several density gradients were tested for the isolation of parasitic nematode, Angiostrongylus costaricensis, first-stage larvae from rodent feces. With a 45/72% Percoll gradient, 83-99% (89.56+/-6.57%) of the larvae were recovered in a clean preparation.
This report describes the clinical and epidemiological findings of an outbreak of histoplasmosis in Brazil involving four persons of the same family, one adult and three children. Contamination by fecal material of bats in the basement of the house was found to be the most likely cause of this outbreak. None of the patients had evidence of immune deficiency. The adult died of acute respiratory distress syndrome. A more severe presentation of the disease in this patient may be partly explained by a heavier exposure to the feces of bats.
This study investigates the relation of fecal production and dietary habits and life-styles in four 21 to 22 year-old healthy female students. The survey was conducted over 30 days and was repeated twice. All feces that were discharged were collected and weighed. The subjects performed very little physical exercise. The fecal weight, the number of defecations per day, gastrointestinal symptoms, feeling of incomplete defecation and of abdominal distention were recorded. The fecal weight was converted to autocorrelation, and the day-by-day variation was examined by a time series analysis (correlogram). Free access to foods was allowed. The weight of each food item was weighed for nutritional evaluation. The daily number of steps walked and sleeping hours were taken as indicators of life-style. The average fecal weight ranged from 96.8 g/day to 127.8 g/day, with a grand mean for the four subjects of 94.1 g/day. The average number of times of defecation during the 60 days period was 53 to 72, or 0-3 a day. The subjects tended to have feeling of incomplete defecation when the stool was hard and fecal weight was less than 100 g per day, whereas the subjects felt incomplete defecation less frequently when the stool was well-formed or pasty. The time series analysis by correlogram indicates that the variation in fecal weight formed a 3-4 day cycle and that the cycle was irrelevant to fecal weight. This survey shows that there was no apparent correlation among the fecal weight and nutrient intake, the number of meals per day, the number of steps walked or sleeping hours per day. It also indicates that defecation factors differ from individual to individual.
A procedure has been established for screening Escherichia coli O157:H7 from bovine feces. It consists of four steps: enrichment, selective culture, phenotyping and genotyping. Modified trypticase soy broth (mTSB) containing 20 microg/mL of novobiocin was used for the enrichment step. The selective culture step was done using sorbitol MacConkey agar containing 0.05 microg/mL of cefixime and 2.5 microg/mL of potassium tellurite. The phenotyping steps include: species confirmation as E. coli, serotyping to O157 and H7, and the detection of verocytotoxin (VT) production. Genotyping was made up of confirmation of the presence of the O157 antigen (rfb), the H7 antigen (ficC), the adherence factor (eaeA), hemolysin (hlyA), and VT production (vt I and vt II) by polymerase chain reaction. mTSB gave better enrichment for E. coli O157 than gram-negative (GN) broth. The detection limit for this screening procedure was 0.85 +/- 0.5 cfu/g. Using the screening procedure described above, E. coli O157 was found in four out of 3062 (0.13%) fresh bovine fecal specimens, and in two out of 78 (2.56%) dairy herds in Taiwan. Three out of the four detected strains were vt II producing E. coli O157:H7, while the other was E. coli O157:H7 which lacked VT-producing ability.
Giardia lamblia cysts in fecal specimens are detected by conventional morphological methods. The direct immunofluorescence monoclonal antibody assay (DFA) is also applied to detect G. lamblia cysts in feces, but little is known about the usefulness of DFA in fecal specimens stored under various conditions. The aim of the present study was to evaluate the DFA for detection of G. lamblia cysts and to compare these results with the direct smear method in long-term storage of non-fixed fecal specimens. Fecal specimens with G. lamblia cysts were stored in a refrigerator at 5 degrees C(14 samples), a freezer at -20 degrees C (9 samples), or in 3.9% formalin-saline solution at room temperature (28 samples). G. lamblia cysts were detected by DFA in all stored specimens, while they were detected in only 56% of refrigerated and 93% of frozen specimens by the direct smear method. The storage period of all samples testing negative by direct smear was more than 24 months. Many degenerated cysts were recognized by DFA when cysts were negative by the direct smear method. Our results indicate that DFA is a useful method for detecting G. lamblia cysts in fecal samples that have undergone long-term storage.
Sulfamethazine was administered to 8- to 10-week-old turkey poults intravenously (IV) at the dose level of 71.5 mg/kg of body weight, orally at the dose level of 143 mg/kg of body weight, or in the drinking water at the concentration of 0.1% over a 6-day period. The concentrations of free sulfamethazine in blood, muscle, skin, kidney, and liver were determined and semilogarithmic plots of concentration vs time for the various tissues indicated that the curve had a linear portion within the first 72-hour period of drug withdrawal. The rates of disappearance of sulfamethazine from the various tissues were proportional to the concentration in the tissues. After 72 hours of withdrawal and for as long as 14 days, sulfamethazine concentrations in kidney, liver, and skin of turkeys given the drug in the drinking water fluctuated between 0.1 and 0.4 ppm. Only 8.6% of the oral dose (143 mg/kg) and 16.5 to 17% of the IV dose (71.5 mg/kg) were recovered in urine and feces as the parent compound during the initial 72-hour period.
OBJECTIVE: To study the infection condition of adenovirus types in infants with diarrhea in Shenzhen. METHODS: 114 fecal samples from diarrhea infants were tested by PCR. The amplified products were analyzed by Taq I and Rsa I endonucleases. RESULTS: The positive rate of adenovirus DNA in fecal samples was 14% (16/114). The amplified products typing procedure started with Taq I endonuclease to identify the type of adenovirus, 11 cleaved by Taq I yielded two fragments of 191 bp and 110 bp, the positive rate of enteric adenovirus was 9.65% (11/114). Followed by Rsa I cleavage, two amplified products yielded 256 bp and 45 bp fragments (Ad40), the positive rate of Ad40 was 1. 75% (2/114), nine amplified products yielded 211 bp and 90 bp fragments (Ad4l), the positive rate of Ad41 was 7.89%(9/114). CONCLUSIONS: The positive rates of the adenovirus types 40 and 41 were 9.65% in feces of infants with diarrhea, and that of the other types of adenovirus was 4.35%.
The aims of the study were to determine the prevalence of Listeria monocytogenes in the feces of healthy Austrians and to characterize the isolates by various typing methods. Stool specimens from 505 healthy volunteers from the Tyrol were tested for the presence of L. monocytogenes using cold enrichment for 6 months and five different detection methods: conventional plating onto Palcam and Rapid'L.MONO agar, immunomagnetic separation (IMS) followed by conventional plating, enzyme-linked fluorescent immunoassay (ELFA), ELISA, and PCR. L. monocytogenes was isolated by conventional plating from one specimen (0.2%), and a further three were positive on immunomagnetic separation (0.8%). Only one specimen tested positive with ELFA and EIA, although it tested negative by conventional culture, IMS, and PCR. Eighteen of 505 samples were positive by PCR (3.6%), and this included three of the four culture-confirmed specimens. Serotyping, phage-typing, arsenic cadmium, antimicrobial-resistance typing and pulsed-field gel electrophoresis showed that multiple L. monocytogenes isolates from three of the four carriers were indistinguishable. Our data indicate that the Austrian fecal carriage rate is at least 0.8%. In view of a listeriosis incidence of 0.16/100,000 per year, the chances of fecal carriage developing into listeriosis appear to be very low.
OBJECTIVE: To estimate the enteric adenoviruses infection in feces of infants with diarrhea in Shenzhen. METHODS: All 195 fecal samples of infants were tested by PCR. Amplimers were analysed by Rsa I endonucleases and the type of enteric adenovirus identified. RESULTS: The positive rate of adenovirus DNA in fecal samples was 13.33% (26/195) while the prevalence rates of enteric adenovirus type 40, adenovirus type 41 and the other type of adenovirus were 2.56% (5/195), 8.21% (16/195) and 2.56% (5/195) respectively. Fever was the characteristic symptom of adenovirus infection. CONCLUSION: Enteric adenovirus type 40 and 41 were important pathogens which caused infant diarrhea in Shenzhen, with latter the majorty.
Presentation of trimethylthiazoline (TMT, a component of fox feces) to laboratory rats elicits freezing, a prominent behavioral sign of anxiety or fear. The present study investigated the neural basis of this unlearned response. Muscimol, a GABA(A) receptor agonist, was injected (4.4 nmol/0.5 microl) into the bed nucleus of the stria terminalis (BNST) as well as into the amygdala, two brain areas known to be involved in anxiety and fear. Temporary inactivation of the BNST but not of the amygdala significantly blocked TMT-induced freezing. This effect was not caused by an enhancement of motor activity after BNST inactivation. In addition, these results confirm previous studies showing that freezing is possible despite amygdala inactivation. These results, and other findings in the literature, suggest that the BNST is critically involved in unlearned fear, whereas the amygdala is more involved in the acquisition and expression of learned fear.
Antigens in human adult feces related to carcinoembryonic antigen (CEA) were analyzed with respect to their molecular masses, CEA domain compositions, and N-terminal amino acid sequences. By avoiding perchloric acid treatment, new fecal antigens related to CEA were identified. The fecal antigens revealed by Western blot were M(r) 78,000, 70,000, 60,000, 50,000, 44,000, 36,000, 33,000, and 25,000 and a species M(r) less than or equal to 14,000. Unlike native CEA, all of the fecal antigens were very poorly soluble in perchloric acid and did not bind to concanavalin A, suggesting that they had undergone significant deglycosylation in the digestive tract. The major fecal antigens were purified by immunoaffinity chromatography and their N-terminal amino acid sequences determined. FA78, FA60, FA33, and the M(r) less than or equal to 14,000 antigen had the N-terminal amino acid sequence of the CEA N-domain, and FA44 and FA25, the sequence of the CEA A2 domain. The CEA domain compositions of the fecal antigens were investigated by probing them with anti-CEA monoclonal antibodies of known domain specificities. The N-terminal amino acid sequences, immunoreactivities with anti-CEA monoclonal antibodies, and apparent molecular masses of the fecal antigens allowed the following domain assignments (based on CEA as N-A1B1-A2B2-A3B3): FA78, N-A1B1-A2B2-A3B3; FA60, N-A1B1-A2B2; FA44, A2B2-A3B3; FA33, N-A1B1; and FA25, A2B2. The M(r) less than or equal to 14,000 antigen was shown to be the N-domain of CEA or nonspecific cross-reacting antigen. FA36 was assigned the N-AB domain structure of nonspecific cross-reacting antigen. The results suggested that FA78, FA60, FA44, FA33, and FA25 were degradation products (including deglycosylation and proteolysis) of CEA and that FA36 was a degradation product of nonspecific cross-reacting antigen.
Spirochetes found in feces of normal and infected pigs during the eight days preceding the onset of the clinical signs of swine dysentery have been studied using electron and phase contrast microscopy. According to their dimensions, diameter, length and pitch, three morphological groups have been described: small, intermediate and large spirochetes. On the basis of their axial filaments arrangement, eight types have been observed. Compared to the control pigs, there was no increase in the total number of spirochetes in infected pigs, except at day 6 before the onset of clinical signs. However, infected pigs have shown a wide spectrum of different morphological types of spirochetes in comparison to control, uninfected pigs, which showed only the type "1-2-1" spirochete of axial filaments arrangement. This last type was predominant in infected swine during the entire incubation period as compared to other spirochete types described.
Shiga toxin-producing Escherichia coli (STEC) are a public health concern. Bacterial culture techniques commonly used to detect E. coli O157:H7 will not detect other STEC serotypes. Feces from cattle and other animals are a source of O157:H7 and other pathogenic serotypes of STEC. The objective of this study was to estimate the pen-level prevalence of Shiga toxins and selected STEC serotypes in pre-slaughter feedlot cattle. Composite fecal samples were cultured and a polymerase chain reaction (PCR) was used to detect genes for Shiga toxins (stx1 and stx2) and genes for O157:H7, O111:H8, and O26:H11 serotypes. Evidence of Shiga toxins was found in 23 pens (92%), O157:H7 in 2 (8%), O111:H8 in 5 (20%), and O26:H11 in 20 (80%) of the 25 pens investigated. Although pen-level prevalence estimates for Shiga toxins and non-O157 serotypes seem high relative to O157:H7, further effort is required to determine the human health significance of non-O157 serotypes of STEC in feedlot cattle.
A small number of colorless colonies grew from DHL agar of feces culture taken as part of a complete physical for a 42 year-old woman who had lived in Singapore for several years. When cultured for first-stage identification using conventional biochemical tests in tubes for Enterobacteriaceae, such as TSI agar slant and SIM medium, the results for lactose reaction (-), saccharose reaction (-), gas (very weak +) and motility (-) were obtained, and Shigella spp. was suspected. Serological tests (by serotype) for Shigella spp. were then conducted. As a result, clear C14 agglutination was observed. Based on these results, the isolate was strongly suspected to be Shigella boydii serotype 14, but since the woman had no symptoms of abdominal pain, diarrhea or fever, such identification was still questionable. When further identification test was carried out using Walk-Away 96 and VITEK 2, non-Shigella spp. identification results were obtained. In second-stage identification, xylolytic activity, acetate salt utilization and use of carbon sources in CA (citrate-acetate) medium were checked, all results were positive, and the isolate was ultimately identified as Inactive Escherichia coli. While Shigella spp. and E. coli are taxonomically similar, they are quite different from each other in terms of pathogenicity. Accurate and rapid identification of Shigella spp. is therefore important.
In this paper, a new, simple and rapid method for the analysis of steroids by reversed-phase high performance liquid chromatography (HPLC) coupled with evaporative light scattering detector (ELSD) is presented. A good resolution of the 12 steroids is obtained within 35 min with a gradient elution using a binary mobile phase composed of methanol and water. The analytical column used was 2.0mm x 150mm, packed with BDS (particle size 5microm) and the mobile phase flow rate was 0.2mL/min. Nebulizer gas flow rate and drift tube temperature, the two critical parameters for determining the reproducibility and signal-to-noise ratio of ELSD were 2.00mL/min and 95 degrees C, respectively. The response of the ELSD was non-linear as a result of basic light-scattering principles, the calibration curve by plotting lnY vs lnX (Y, X stand for peak area and concentration, respectively) shows a good linearity in the range of 5-50mg/L with a correlation coefficient higher than 0.99 for individual steroid and with relative standard deviation less than 6.92%. This method has been applied to the determination of steroids in milk powder and birds' feces, solid phase extraction on reverse phase bonded C18 (5mL/500mg) was used to treat the samples efficiently. The recoveries of the steroids were between 84.0%-102.8%.
Some serotypes of Yersinia enterocolitica might cause diarrheas and/or invasive infections. The aim of this work was to analyze virulence factors and associated markers in a strain of Y. enterocolitica isolated from human diarrheic feces. The strain analyzed was included in the biotype 1A. The virulence markers determination as well as the search of the genes vir F and ail, were negatives. However, it was demonstrated enterotoxin production at 20 degrees C, and at 37 degrees C in osmolarity conditions and pH similar to the human intestine. The enterotoxin presented reactivity for the infant mouse test, although it could not be proven the presence of yst gene by PCR. The results obtained by us, coincident with those of other investigators, indicated that certain clinical isolates of Y. enterocolitica of the biotype 1A ("avirulent"), could be the etiological agent of the illness through other mechanisms of virulence, that would differ from those previously characterized in species of enteropathogenic Yersinia.
The numerous specimens of liquid diarrheas received at the clinical parasitology laboratory of "Pedro Kouri" Institute of Tropical Medicine were studied searching for the presence of pathogenic agents of Coccidium subclass and of the microspore family. The conditions to diagnose microsporidia were created on July 13, 2000, and the method was incorporated to the battery of diagnostic techniques used in the laboratory. A cut-off was made after 4 months to analyze the positivity found. A frequency distribution study was conducted and it was observed that of the 170 studied samples, 51 proved to be positive for some protozoon, which represents 30% of positivity. The most frequently found species were Cryptosporidium parvum and microsporidium. Association of various species in a same sample was found in 13.1% of the positive samples. The most frequent associations were: Cryptosporidium parvum and Cyclospora cayetanensis, Cryptosporidium parvum and microsporidium, and Cyclospora cayetanensis and microsporidium, each with 2 patients. Association of 3 different species in a same sample was also found. All the individuals in the series were HIV seropositive and the most affected had less than 200 TCD4+ lymphocytes/mL. This was the first report of microsporidia in feces in Cuba. It was possible after applying and interpreting the technique previously described to identify this phylum. As there is a trend towards the increase of cases with HIV/AIDS infection, it is necessary that the laboratories have the indispendable reagents to perform the modified Didier's trichromic staining and that the professionals and technicians of the parasitology laboratories of the country be trained in the procedures for identifying and recognizing these opportunistic protozoa.
We compared the overall usage of veterinary therapeutic antimicrobials in Japan to the proportion of antimicrobial-resistant. Escherichia coli isolated from the feces of apparently healthy food-producing animals in 2001. The annual sales volume of veterinary antimicrobials, which is published and accumulated information, was subdivided according to the target animal species (cattle, pigs, broiler chickens, and layer chickens). The antimicrobial susceptibility of the isolates was examined against 7 classes of 11 antimicrobials. The rates of antimicrobial resistance among the isolates were found to correlate significantly with the usage of antimicrobial agents in cattle, pigs, and broiler and layer chickens. Therefore, the overall usage of veterinary antimicrobials appears to contribute to the appearance of antimicrobial resistance in E. coli isolates from apparently healthy food-producing animals.