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[Cryptosporidium parasitism in children with acute diarrhea].

During the period from August 1987 to July 1990, 241 fecal samples collected from 1 to 48 months old children with acute diarrhea and examined in the Child's Institute of Hospital das Clínicas of Faculdade de Medicina da Universidade de São Paulo, were submitted to parasitological tests in the Enteroparasitosis Section of the Instituto Adolfo Lutz. Fourty two (17.43%) fecal samples showed Cryptosporidium sp. oocysts by carbol-fucsin stain. Parasitism by Cryptosporidium sp. was more frequent during the period comprised from March to May, in the studied three years. The authors discussed the Cryptosporidium sp. in association with other agents.

Acute Disease↗

Isolation of picornavirus from feces and semen from an infertile bull.

Virus was isolated from semen and fecal samples from a bull with orchitis, testicular degeneration, aspermatogenesis, and loss of libido. Both isolates were classified as picornavirus, bovine enterovirus serotype I, on the basis of physical, chemical, and serologic characteristics. Veterinary practitioners that may suspect viral infection as a cause of bovine infertility should submit both semen and fecal samples for virus isolation and identification.

Animals↗

Zinc absorption estimated by fecal monitoring of zinc stable isotopes validated by comparison with whole-body retention of zinc radioisotopes in humans.

Knowledge about zinc availability from human diets is limited due to methodological difficulties. Recently developed stable isotope techniques for estimating dietary zinc absorption were compared with radioisotope techniques in five men and three women. Stable and radioactive zinc isotopes were simultaneously administered. Fecal excretion of the isotopes as well as whole-body retention of the radioactive zinc isotope was monitored. Concentration of stable zinc isotope label in fecal samples was determined by inductively coupled plasma mass spectrometry by fully quantitative measurements and from inductively coupled plasma mass spectrometry isotope ratios combined with analysis of total zinc content using atomic absorption spectrometry. Zinc absorption estimated from whole-body retention was 27 +/- 6% (mean +/- SD), estimated zinc absorption obtained by fecal monitoring of radioisotope was 26 +/- 9%, and the two stable zinc measurements resulted in values of 29 +/- 12 and 33 +/- 12%, respectively. There was no significant difference in zinc absorption estimated from whole-body retention and with the fecal monitoring methods. Recovered stable zinc isotope label was significantly lower than recovered radioisotope. For individual fecal samples, systematic differences of 16% and 12%, respectively (P < 0.05), between the radioisotope recovery and the recovery of stable isotopes with the two methods for measurement was observed. The stable zinc isotope technique for measurement of zinc absorption resulted in mean results similar to those of the radioisotope technique, but with a larger variation in the measurements.

Adult↗

Rapid detection of human group C rotaviruses by reverse passive hemagglutination and latex agglutination tests using monoclonal antibodies.

Reverse passive hemagglutination (RPHA) tests and a latex agglutination test with monoclonal antibodies (MAbs) were developed for the rapid detection of noncultivatable human group C rotaviruses. For RPHA tests, two MAbs, MAb 5A12 recognizing the outer capsid and MAb 13A3 recognizing the inner capsid, were separately used for the coating of sheep erythrocytes (SRBCs). Forty-six fecal samples were examined to confirm the practicality of the tests. As a result, there was concordance between the RPHA test with SRBCs coated with MAb 5A12 and polyacrylamide gel electrophoresis of viral RNA (RNA-PAGE) in 44 (95.6%) of 46 samples, while the diagnoses by the RPHA test with SRBCs coated with MAb 13A3 were in complete agreement with those by RNA-PAGE. Furthermore, a latex agglutination test with MAb 13A3 was also developed, and this test was fast enough and sensitive enough to successfully detect the viruses from most fecal samples within 2 min. The present procedures would be useful for the diagnosis of human group C rotavirus infections in clinical laboratories which are not well equipped.

Antibodies, Monoclonal↗

Intestinal parasitic infection in children in post-disaster situations years after earthquake.

BACKGROUND: Two earthquakes in the north-west region of Turkey destroyed 80% of the houses and schools in Duzce in 1999. This study was conducted to determine the parasitic infection rate associated with the post-earthquake unhealty living conditions and related epidemiological risk factors. METHODS: Two populations living and studying in different socioeconomic conditions as a result of the earthquake were compared: Group 1 (study group) consisted of 326 children living and studying in transitory houses and classes. Group 2 (control) consisted of 127 children living in normal houses and studying in normal school classes. Fecal samples were obtained from both groups and examined for intestinal parasites' eggs and trofozoid forms. In addition, selotype procedure was applied to both populations. Epidemiological data determining the socioeconomic status of the populations were collected by questionnaire. RESULTS: In group 1, Giardia lamblia (G. lamblia) cysts were observed in 10.4% of the fecal samples and Enterobius vermicularis (E. vermicularis) eggs were observed in 13.5% of selotype samples. In group 2, Giardia cysts were observed in 3.1% of the samples and E. vermicularis eggs were observed in 5.5% of selotype samples. The rate of Giardiasis and Enterobiasis was found to be significantly higher in children still living and studying in temporary houses and schools years after the earthquakes (P < 0.05). The following pidemiological and social factors arising after the earthquakes were associated with increased rate of G. lamblia and E. Vermicularis infections: number of communal toilets per child at school, socioeconomic level of the children, dimensions of the classrooms where the children are studying and living and frequency of hand-washing at school. These parameters are significantly different between the groups (P < 0.05). CONCLUSION: Giardiasis should be considered as an emerging disease in postdisaster situations and adequate prevention measures should be implemented in these circumstances. It should also be known that the rate of Enterobiasis is increased in populations living in crowded unhealthy conditions.

Adolescent↗

Imipenem concentrations in colorectal surgery and impact on the colonic microflora.

Twenty patients undergoing colorectal surgery were given, as prophylaxis, imipenem-cilastatin intravenously. Ten of them received a dose of 0.5/0.5 g of imipenem-cilastatin at induction of anesthesia, followed by subsequent doses of 0.5/0.5 g every 6 h for 48 h. The other 10 patients were given 1.0/1.0 g imipenem-cilastatin in the same way for 48 h. Samples from serum, intestinal mucosa, and feces were taken for analysis of imipenem concentrations during the day of surgery. The mean concentrations in serum at 1 h after the first imipenem dose were 15.9 +/- 1.7 micrograms/ml for the 0.5-g dose and 68.2 +/- 8.2 micrograms/ml for the 1.0-g dose. The mean half-lives were 1.5 and 1.4 h, respectively, and the mean areas under the serum concentration-time curve were 41.2 +/- 6.0 and 128.3 +/- 13.5 mg.h/liter, respectively. The imipenem concentrations in the intestinal mucosa varied between less than 0.1 and 3.6 mg/kg for the 0.5-g dose and 3.2 and 13.4 mg/kg for the 1.0-g dose. The concentrations in the fecal samples varied between less than 0.1 and 5.0 mg/kg for the 0.5-g dose and 0.7 and 11.3 mg/kg for the 1.0-g dose. Fecal samples were also collected during the investigation period for cultivation of aerobic and anaerobic bacteria. The aerobic bacteria--staphylococci, streptococci, enterococci, and enteroaerobic enterococci, and enterobacteria--were and anaerobic bacteria. The aerobic bacteria--staphylococci, streptococci, enterococci, and enterobacteria--were suppressed significantly during the imipenem prophylaxis period. Among the anaerobic bacteria, cocci, bifidobacteria, eubacteria, lactobacilli, clostridia, fusobacteria, and bacteroides decreased markedly during the same period. The microfloras were normalized after 2 weeks. There were no differences between the patients receiving 0.5-g doses of imipenem and those receiving 1.0-g of imipenem. No postoperative infections occurred.

Adult↗

Hygiene habits and carriers in families with a child who has had typhoid fever.

The relationship between asymptomatic shedding of bacterial enteropathogens and the hygiene habits of families who have had a child with typhoid fever (TF) are investigated. The sample was made up of 80 families: 40 families in which one child had had TF (Group A) and 40 in which no children or either of the parents had had a history of TF (Group B). In each group 20 families belonged to a low socioeconomic status (SES) and 20 to a high SES. A structured interview was used to evaluate the SES and the hygiene habits of the child; observations were made to measure the hygiene habits of the family (toilet, kitchen and food preparation) and bacteriological studies (fecal samples and hand markers). Results show that carriers were more frequent in Group A than in Group B. The bacterial species found were significantly more numerous in Group A than in Group B (fecal samples: E. coli, the classic serotypes, Shigella ssp, and hand markers: E. coli). Families of Group A had higher carriage rates than those of Group B. Finally there exists a significantly higher association between inadequate hygiene habits and carrier families. These results show the need to teach specific habits of proper hygiene to the entire population, because the fact of belonging to the high SES does not in itself preclude inadequate hygiene habits.

Adolescent↗

Microarray method to monitor 40 intestinal bacterial species in the study of azo dye reduction.

Azo dyes are widely used in dye manufacturing, paper printing, textile industries, and as tattoo pigmentation. Since intestinal and skin bacteria can metabolize certain azo dyes to carcinogenic compounds, many researchers have studied the azoreductases of these bacteria. In this study, we used a microarray method to identify the intestinal bacterial species from cultured fecal samples in Brain Heart Infusion (BHI) broth with or without azo dyes that may be involved in azo dye reduction. The microarray was designed to identify 40 bacterial species that are reported in the literature to be predominant in human feces. Results from this study showed 26-30 species are present in the cultured fecal samples. The representative bacteria were then examined for the azo dye reduction activity.

Azo Compounds↗

Evidence of exposure of waterfowl and other aquatic birds to the hemagglutinating duck adenovirus identical to EDS-76 virus.

Serum and fecal samples from 12 species of aquatic birds were studied for evidence of exposure to a hemagglutinating duck adenovirus (DAV). DAV is serologically indistinguishable from egg-drop syndrome-76 virus. A total of 285 serum samples were tested by the hemagglutination-inhibition (HI) test. Forty-two percent of the birds had HI antibodies, with titers ranging from 8 to 256. Wild ducks showed the highest frequency of antibodies (56%) while in coots and grebes, antibody was less frequent, 33% and 26%, respectively. Attempted virus isolations from 79 fecal samples were unsuccessful. The data support the hypothesis that DAV is indigenous in wild duck populations and suggest that infection and viremia are limited in time and occur at a very early age.

Adenoviridae↗

[Diagnosis of Lawsonia intracellularis using the polymerase chain reaction (PCR) in pigs with and without diarrhea and other animal species].

Lawsonia (L.) intracellularis, an obligately intracellular bacterium, causes proliferative enteropathy (PE) in swine and, occasionally, in other animals. To determine the spread of the agent among German pig herds pooled fecal samples of five animals each of clinically normal Hessian pig herds collected between november 1998 and february 1999 as well as feces (n = 1684) from individual animals representing 648 herds, sent to our laboratory by veterinarians from all parts of Germany, were tested for L. intracellularis using the polymerase chain reaction (PCR). In addition, fecal samples from diarrhoic foals (n = 46), dogs (n = 57), cats (n = 50), calves (n = 37), hedge hogs (n = 9), seals (n = 8) and one giraffe were also studied. DNA was extracted from feces using high concentrations of chaotropic salt and diatomaceous earth. For PCR, primers flanking a 279 bp fragment of L. intracellularis DNA were used (JONES, G. F., WARD, G. E., MURTAUGH, M. P., LINN, G. (1993), J. Clin. Microbiol. 31, 2611-2615). Amplificates were separated by agarose gel electrophoresis and visualized under UV-light. L. intracellularis was found in 26 (12.8%) samples from 21 (30.0%) of the Hessian pig herds without symptoms of diarrhoea. In feces of pigs with diarrhoea (n = 1684) the agent was present in 431 (25.6%) samples originating from 224 (34.6%) herds. Of the other animal species studied, L. intracellularis was detected in feces of 4 (7.0%) dogs, 2 (5.4%) calves, 3 (33.3%) hedge hogs and in the sample of the giraffe. The remaining species were all tested negative.

Animals↗

[Evaluation of an immunoenzyme technique for the detection in feces of the Giardia intestinalis antigen].

An evaluation is made of the utility for the diagnosis of giardiasis of an enzyme immunoassay (EIA) that detects the GSA65 specific antigen in feces, as compared with the formalin-ether sedimentation test. Within the context of a longitudinal study on intestinal parasitization, 147 fecal samples from children attending 7 day care centres in the city of Salamanca were studied. The feces were examined a few hours after collection by the sedimentation technique and conserved in 10% formol. They were later studied by the EIA method. G. intestinalis was detected by microscopy and/or EIA in 26 of the 147 fecal samples (17.7%). Both assays were positive in 22 of the 26 cases detected (84.6%) and the remaining 4 cases were positive only by EIA (4/26; 15.4%). The EIA assay, using visual reading and very simple to implement, showed sensitivity, specificity, positive predictive and negative predictive values of 100, 96.9, 84.6 and 100%, respectively.

Animals↗

Four cases of hemolytic uremic syndrome--source contaminated swimming water?

In June '93, 4 children, aged 1.5-3.5 years, all living in one town, were admitted to our hospital with the diagnosis hemolytic uremic syndrome (HUS) within one week. In cooperation with the local health authorities a common source was searched for. Questionnaires indicated that the single condition shared by all patients was swimming water. The patients were not acquainted, visited different daycares, and had no food resources in common. All 4 patients bathed in the same, shallow, recreational lake within a period of 5 days. During this time the air temperature was high according to Dutch standards (around 27 degrees C), and many people visited the lake, estimated several hundreds a day. The water level was lower than normal. Diarrhea followed 3-11 days after swimming and the first clinical symptoms of HUS developed 6-7 days after the onset of diarrhea. The lake was closed for swimming when the fourth HUS patient was diagnosed and the possibility of transmission by way of the lake was mentioned. E. coli O157: H7 was demonstrated in the fecal samples of 2 index patients. The samples were taken 9-20 days after the start of diarrhea. Antibodies to O157 and verotoxin 2 were strongly positive in all patients. A local outbreak of diarrheal illness was not registered. Of 16 family members who also swam in the same lake, 7 developed symptoms of enteritis, 3 had positive cultures of their fecal samples and 5 had positive serology. Pulsed-field gel electrophoresis of the E. coli isolates of the patients and family members showed an identical pattern. No O157: H7-DNA could be detected in filter concentrated lake water samples using polymerase chain reaction (PCR) enhancement. These samples were, however, taken 16 days after the latest possible date of contamination of our patients, 15 days after decrease of the air temperature to 15-17 degrees C, and 14 days after the inlet from water from the environment. It could thus very well be that the microorganism was no longer present. This third report of swimming water associated HUS should direct environmental surveys in similar cases of local HUS outbreaks.

Antibodies, Bacterial↗

Transmissible drug resistance in Escherichia coli isolated from poultry and their carcasses in Iran.

A high prevalence of multiple antibiotic resistance was observed in populations of Escherichia coli isolated from fecal samples and the carcasses of poultry in a University farm. Almost all of the cultures were resistant to tetracycline, streptomycin, ampicillin and sulfoamide. Association between antibiotics used for prophylaxis or growth promotion and isolation of E. coli with resistance to these antibiotics was observed. One hundred of 648 (15.43%) of resistant cultures of E. coli isolated from fecal samples and the carcasses were capable of transferring resistance determinants to a laboratory strain of E. coli K-12.

Animals↗

Potential risk factors for Cryptosporidium infection in dairy calves.

Fecal samples from 7,369 calves on 1,103 farms were examined for cryptosporidia in a nationwide survey, using monoclonal antibody technique. Cryptosporidium oocysts were found in calves from 652 (59.1%) of the farms and in 1,648 (22.4%) of the tested calves. Almost half the calves between 7 and 21 days of age had cryptosporidia in their fecal samples. Prevalence was highest during the summer. Farms with multiple-cow maternity facilities and farms with > 100 milking cows were the most likely to have calves with cryptosporidia.

Age Distribution↗

Survey for internal parasites in cattle in Kentucky (1993).

Fecal samples were examined between 17 September and 1 November 1993 from 1765 cattle (one bull, 533 cows, 474 heifers, 22 steers, and 735 calves) on pasture on 15 farms in 11 counties in Kentucky for eggs or larvae of internal parasites. All of the cattle were beef-type except for 22 which were dairy-type. In fecal samples from the bull, cows, heifers, steers, and calves, the types of helminth eggs present were trichostrongyles (excluding Nematodirus) in 0%, 25%, 31%, 86%, and 93%, Nematodirus in 0%, 0%, < 1%, 0%, and 34%, Strongyloides in 0%, 0%, < 1%, 0%, and 7%, Trichuris in 0%, 0%, 0%, 0%, and 2%, Capillaria in 0%, 0%, 0%, 0%, and < 1%, and Moniezia in 0%, 1%, 8%, 5%, and 21%, respectively. Dictyocaulus viviparus larvae were present in feces of calves (7%) on one farm and heifers (< 1%) on another farm.

Animals↗

Dietary fiber differentially alters cellular fatty acid-binding protein expression in exfoliated colonocytes during tumor development.

We investigated the utility of noninvasive technology utilizing feces containing exfoliated colonocytes to determine whether changes in fecal fatty acid-binding proteins have predictive value in monitoring the neoplastic process. Ninety male Sprague-Dawley rats were randomly divided into four groups in a 2 x 2 factorial design, with two dietary fiber sources (wheat bran or oat bran) and two treatment groups (injection with a carcinogen, azoxymethane, or saline). Fresh fecal samples were collected at Week 16 postinjection, and tumor frequency was determined at Week 36 of the study. Semiquantitative "mimic" reverse transcriptase polymerase chain reaction was used to quantitate the expression of liver fatty acid-binding protein (L-FABP), intestinal fatty acid-binding protein (i-FABP), and acyl CoA-binding protein (ACBP) mRNA in fecal samples to establish their prognostic value. Rats fed wheat bran diets had a lower incidence of tumors (p < 0.05). There was no effect of carcinogen injection or tumor incidence on the expression of L-FABP, i-FABP, or ACBP mRNA, L-FABP and i-FABP mRNA expression were significantly higher (p < 0.05) in feces from animals fed a wheat bran diet than in feces from animals fed an oat bran diet. In contrast, the expression of ACBP mRNA was significantly lower (p < 0.05) in animals fed a wheat bran diet than in animals fed an oat bran diet. Wheat bran also increased (p < 0.05) the total excretion of L-FABP, i-FABP, and ACBP over a 48-hour period. These data suggest that exfoliated colonocyte fatty acid-binding protein mRNA status may provide insight into the mechanisms by which diet influences colonic physiology.

Animals↗

[Detection of enteroaggregative Escherichia coli from sporadic diarrhea patients].

A study of enteroaggregative Escherichia coli (EAggEC) which was recently reported as a causative agent of diarrhea was attempted, by isolating these organisms from the fecal samples collected from sporadic diarrhea patients in Miyazaki Prefecture during the period from January 1993 to April 1998, and by investigating several characteristics of the isolates. By using the PCR method targetting aggR gene which was a transcriptional activator of aggregative adherence fimbria I expression, thirty four strains of aggR(+)-Escherichia coli (E. coli) were detected from 2,652 fecal samples. Twenty nine of these 34 isolates were confirmed as EAggEC by demonstration of aggregative adherence to HEp-2 cells, and the other 5 isolates were not EAggEC because they showed negative adherence to HEp-2 cells. The above mentioned, aggR-PCR method revealed that there were a few non-EAggEC strains with aggR gene. It has been reported that aggregative adherence fimbriae are encoded by the plasmid of about 60 Md. All of the 29 EAggEC isolates possessed plasmids of about 50 Md or more, and these plasmids were suggested to relate to aggregative adherence fimbriae. Sixteen (55%) of the 29 isolates were classified serologically into two serotypes, O111:H21 and O126:H27, and the other 13 isolates were classified into ten groups or more which included a few strains in a group. EAggEC heat-stable enterotoxin 1 (EAST1) gene was demonstrated in nineteen of 29 isolates. In drug susceptibility test, 72%, 59% and 21% of the 29 isolates showed resistance to Ampicillin, Cefazolin, and Streptomycin, respectively.

Bacterial Adhesion↗

[Isolation and identification of Cryptosporidium from various animals in Korea. III. Identification of Cryptosporidium baileyi from Korean chicken].

Each of SPF chicken (Hi-Line strain, 2-day-old males) was inoculated with 2.5 or 5 x 10(4) oocysts by stomach tube. The oocyst was the medium type of Cryptosporidium previously isolated from Korean chicken origin, and passed in 2-day-old SPF chicken. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of the developmental stages, the bursa of Fabricius of the chicken was examined by transmission electron microscopy (TEM) on the 12th day postinoculation. The prepatent period for 8 chicken was 5.9 days postinoculation on the average, and the patent period was 12.9 days. The number of oocysts discharged per day for the chicken was reached peak on day 12 postinoculation on the average. A large number of oocysts was found in fecal samples obtained from inoculated chicken on days 8-14 postinoculation. The ultrastructural feature of almost every developmental stage of the medium type from chicken was very similar to that of Cryptosporidium previously isolated from mammalia including human and birds except for the attachment site of C. muris to the mucus cell from mammalia, but dimension of the oocysts from fecal samples of the medium type was different from those of C. meleagridis and mammalia origin. The above results reveal that the medium type of Cryptosporidium of Korean chicken origin is identified as Cryptosporidium baileyi.

Animals↗