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Third generation cephalosporin-resistant gram-negative bacilli in the feces of hospitalized children.

In view of the widespread use of third generation cephalosporins in hospitalized infants, we attempted to determine whether their use was associated with the emergence of resistance in fecal Gram-negative bacilli. Stools from infants hospitalized for varying durations were cultured on MacConkey agar containing 4 micrograms/ml of cefotaxime. All isolates growing on this medium were identified and their susceptibilities to 29 antimicrobial agents were determined. Sixty-five infants were studied of whom 44 were receiving a third generation cephalosporin, 7 another antibiotic and 14 no antibiotic. Thirty-one strains resistant to third generation cephalosporins (minimal inhibitory concentrations > or = 16 micrograms/ml) to cefotaxime, ceftriaxone or ceftazidine) were isolated from 26 infants. The proportions of infants with resistant strains were not significantly different whether they were: (1) receiving a third generation cephalosporin or not; (2) hospitalized for longer or shorter than 2 days or not; (3) older or younger than 3 months or not. Notably 8 infants harbored resistant strains within 24 hours of admission. The commonest resistant strains isolated belonged to the genera Enterobacter (10), Citrobacter (6), Serratia (3), Cedecea (3) and Chromobacterium (3). In conclusion hospitalized infants had a high incidence of fecal colonization with Gram-negative bacilli resistant to third generation cephalosporins. These bacteria were predominantly those known to produce broad spectrum beta-lactamases. This colonization was not necessarily associated with the infant receiving such antibiotics or with prolonged hospitalization.

Anti-Bacterial Agents↗

Phylogeny of Helicobacter isolates from bird and swine feces and description of Helicobacter pametensis sp. nov.

Previously, nine fecal isolates from wild birds and a domestic swine were identified as helicobacters by phenotypic characterization and reaction with a helicobacter genus-specific DNA probe. These isolates fell into three biotypes by analysis of phenotypic traits. To further characterize these isolates, full 16S rRNA sequences were determined for strains representing each biotype, and sequence comparison indicated that the strains represented three novel, phylogenetically defined Helicobacter species. Three 16S rRNA-based DNA probes were designed and used to identify the remaining strains. Probe reactivity divided the strains into the same three groups identified phenotypically. Six of the isolates represented a new species of the genus Helicobacter for which we propose the name Helicobacter pametensis sp. nov. The following phenotypic features distinguished H. pametensis from other Helicobacter and Campylobacter species: positive tests for oxidase, catalase, alkaline phosphatase, nitrate reduction, growth at 42 degrees C, and growth in the presence of 1% glycine; negative tests for urease, gamma glutamyl transpeptidase, indoxyl acetate hydrolysis, and hippurate hydrolysis; and susceptibility to nalidixic acid and cephalothin. H. pametensis cells were motile and possessed one subterminal sheathed flagellum at each end. The two additional Helicobacter species were similar to H. pametensis except that they were urease positive, hydrolyzed indoxyl acetate, and were resistant to cephalothin. Because these two additional species are phenotypically similar and are represented by only two isolates for one species and one isolate for the other, they are not formally named but are referred to as Helicobacter sp. "Bird-B" and Helicobacter sp. "Bird-C."(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Transformation of bile acids by mixed microbial cultures from human feces and bile acid transforming activities of isolated bacterial strains.

Microbiol transformation of cholic acid and chenodeoxycholic acid by anaerobic mixed cultures of human fecal microorganisms was investigated, and the results were examined in relation to the bile acid transforming activities of 75 bacterial strains isolated from the same fecal cultures. The reactions involved in the mixed cultures were dehydrogenation and dehydroxylation of the 7 alpha-hydroxy group in both primary bile acids and epimerization of the 3 alpha-hydroxy group in all metabolic bile acids. Extensive epimerization of the 7 alpha-hydroxy group of chenodeoxycholic acid yielding ursodeoxycholic acid was also demonstrated by certain fecal samples. 7 alpha-Dehydrogenase activity was widespread among the fecal isolates (88% of 16 facultative anaerobes and 51% of 59 obligate anaerobes), and 7 alpha-dehydroxylase activity was revealed in one of the isolates, and unidentified gram-positive nonsporeforming anaerobic bacterium. 3 alpha-Epimerization was effected by seven strains assigned to Eubacterium lentum, which were also active for 3 alpha- and 7 alpha-dehydrogenations. No microorganism accounting for 7 alpha-epimerization was recovered among the isolates. Splitting of conjugated bile acid was demonstrated by the majority of obligate anaerobes but the activity was rare among facultative anaerobes.

Bacteria↗

Enzyme-linked immunosorbent assay employing monoclonal antibodies for direct identification of enteric adenoviruses (Ad40,41) in feces.

For detection and identification of enteric adenovirus (Ad) types 40 and 41 in stool specimens, enzyme-linked immunosorbent assay (ELISA) was developed with the use of three monoclonal antibodies: Ad group-specific, Ad40 type-specific, and Ad41 type-specific antibodies. Of 860 fecal samples from patients with acute gastroenteritis, 44 strains of Ad were isolated using Graham 293 cell cultures. Of these isolates, 20 were typed as Ad40, 18 were Ad41, and 6 were other Ads by neutralization tests with cell cultures. Results of the ELISA tests on these 860 fecal samples resulted in good agreement to those with the cell culture method. The ELISA tests using Ad type-specific monoclonal antibodies proved to be a specific and rapid technique for laboratory diagnosis of acute gastroenteritis caused by enteric Ads.

Acute Disease↗

Prevalence of colonization factor antigens (CFAs) and adherence to HeLa cells in enterotoxigenic Escherichia coli isolated from feces of children in São Paulo.

Fifty-eight enterotoxigenic Escherichia coli (ETEC) strains, isolated from children with and without diarrhea in São Paulo, were examined for the presence of colonization factor antigens (CFAs) and their ability to adhere to HeLa cells. Antisera to CFA/I, the coli surface (CS) antigens CS1CS3, CS2CS3, and CS2 of CFA/II, CFA/III, and CS5CS6 and CS6 of CFA/IV were used. CFAs were identified in 43% of the ETEC strains: 40% of the CFAs strains with CFAs harbored CFA/I, 24% carried CFA/II (CS1CS3), 24% carried CFA/IV (CS6), and 12% carried CFA/IV (CS5CS6). CFAs occurred mainly among ETEC strains producing only heat-stable (ST-I) enterotoxin and in strains also producing heat-labile toxin (LT-I). No ETEC strains tested expressed CFA/III. A marked change in serotypes of ST-I-producing strains was found in São Paulo between 1979 and 1990. Adherence to HeLa cells was detected in 14% of the ETEC strains. All of them had a diffuse adherence pattern and produced only ST-I, and 88% carried CS6 antigen.

Animals↗

Polymerase chain reaction for the detection of Chlamydia psittaci in the feces of budgerigars.

The polymerase chain reaction (PCR) targeting the ompA gene of Chlamydia psittaci was evaluated for its ability to detect chlamydiae in fecal specimens of budgerigars as compared with isolation procedures using cell culture and embryonated egg inoculations. Several procedures for PCR template DNA preparation were compared so as to determine their detection levels for chlamydiae propagated in cell culture in the presence of fecal materials. Tween-20 and proteinase K treatments followed by centrifugation of the template DNA were found to be an appropriate procedure for DNA preparation for primary PCR. Subsequent nested PCR was shown to detect 4.8 IFU/ml or 84 particles/ml of chlamydiae. Chlamydiae in 50 fecal specimens from apparently healthy budgerigars were examined by nested PCR and several other methods. Nested PCR detected chlamydiae at a higher rate (12/50, 24%) than the isolation procedure in embryonated eggs (6/50, 12%). Primary PCR combined with the isolation procedure in cell culture gave a detection rate (5/50, 10%) similar to that of isolation from embryonated eggs. Detection rates by primary PCR (1/50, 2%) and in cell culture (0%) were inferior to the other procedures.

Animals↗

Comparison of four microbial enzymes in Clostridia and Bacteroides isolated from human feces.

The activities of four microbial enzymes (azoreductase, nitroreductase, beta-glucuronidase, and beta-glucosidase) in major anaerobic members of human fecal microflora were quantified and the influence of the host factors on expression of these microbial enzyme activities was also investigated. Clostridium paraputrificum and C. clostridiiforme showed much higher activities than other fecal anaerobes tested. Nitroreductase activity in C. paraputrificum isolated from fecal specimens of patients with colon cancer was significantly (P < 0.05) higher than that in the clostridia isolated from healthy subjects and the subjects given high beef diets. However, the activities of some microorganisms tested showed marked differences in each strain.

Aerobiosis↗