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At least 19 recordsLinked to original sources

Identification of Enterobacteriaceae by the automicrobic system: Enterobacteriaceae biochemical cards versus Enterobacteriaceae-plus biochemical cards.

Enterobacteriaceae Biochemical Cards (EBC) may be used in the AutoMicrobic system for identification of enteric bacilli. Recently, the card has been modified to permit identification of enteric and certain nonenteric bacilli. Also, minor modifications have been in the computer program used for interpretation of tests with the new cards (EBC+). The two types of cards (EBC and EBC+) were tested in parallel and found to be in agreement with 97% of 650 Enterobacteriaceae. Most of the discrepancies were resolved when selected strains were retested on 3 separate days. A lack of absolute reproducibility with either system was demonstrated and explained most of the initial discrepancies. Approximately 97% of the AutoMicrobic system identifications agreed with those obtained from standard reference methods, after equivocal AutoMicrobic system results (P less than 0.80) were excluded. Equivocal responses occurred with 4% of our EBC tests and 7% of our EBC+ tests; additional tests are needed before such strains can be identified with confidence.

Bacteriological Techniques↗

Evaluation of the 3M Petrifilm Enterobacteriaceae Count plate method for the enumeration of Enterobacteriaceae in foods.

Results of the 3M Petrifilm Enterobacteriaceae Count (EB) plate method were compared with those of the standard violet red bile glucose agar (VRBG) method for the detection and enumeration of Enterobacteriaceae. Studies involving 107 bacterial strains demonstrated that the Petrifilm EB plate method is as sensitive as and more selective than the VRBG method. Sixty of the 62 pure Enterobacteriaceae cultures were recovered by both methods. In addition, 38 of the 45 non-Enterobacteriaceae organisms did not grow on the Petrifilm EB plate, while 28 of the 45 non-Enterobacteriaceae organisms did not grow on the VRBG plate. Colony counts from 174 naturally contaminated and 120 artificially inoculated dairy and nondairy food samples showed that the Petrifilm EB plate method performed as well as or better than the standard VRBG method for the enumeration of Enterobacteriaceae.

Animals↗

Problems in the laboratory isolation of simian hemorrhagic fever viruses and isolatse metaboli enterobacteriaceae classificat enterobacteriaceae enzymology hydrogen peroxide metaboli enterobacteriaceae metaboliion of the agent responsible for the Sussex-69 epizootic.

At least six epizootics of simian hemorrhagic fever have occurred at four different primate centers. Although these diseases could easily be transmitted to other monkeys of the Macaca species, difficulty has been encountered in isolating the causative virus in cell culture. The results of this study have shown that the isolation of simian hemorrhagic fever virus strains in cell culture is dependent upon the use of a susceptible MA-104 cell strain and that the ability of such strains to support the replication of these viral agents may vary. By using this information we have been able to isolate a viral agent in cell culture from materials derived from the Sussex/69 epizootic.

Animals↗

3M Petrifilm enterobacteriaceae count plate method for enumeration of enterobacteriaceae in selected foods: collaborative study.

The practice of detecting and enumerating all oxidase-negative, glucose-fermenting-Gram-negative rods (i.e., the family Enterobacteriaceae) is used to indicate unsanitary or inadequate food processing conditions. The objective of this interlaboratory collaborative study was to evaluate and compare the methods described in Standard Methods for the Examination of Dairy Products (SMEDP) and the Compendium of Methods for the Microbiological Examination of Foods (Compendium) with a commercial product, the 3M Petrifilm Enterobacteriaceae Count Plate, for the recovery of Enterobacteriaceae in foods. Six foods--cheddar cheese, milk, flour, frozen prepared meals, frozen broccoli, and nut pieces--were analyzed for Enterobacteriaceae by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test portions consisting of a control test portion and 3 levels of inoculated test portion, each in duplicate. Each test portion was tested by the Petrifilm Enterobacteriaceae Count Plate method as well as the SMEDP or Compendium methods. The precision estimates (repeatability or within-laboratory variation, and reproducibility or between-laboratory variation) were calculated with standard statistical techniques.

Animals↗

Tigemonam activity against clinical isolates of Enterobacteriaceae and Enterobacteriaceae with known mechanisms of resistance to beta-lactam antibiotics.

Tigemonam, a new oral monobactam, was at least as active as aztreonam or carumonam against clinical isolates of Enterobacteriaceae (MIC90:0.06-16 mg/l). Tigemonam was very stable in the presence of classical plasmid mediated beta-lactamases but its MICs were increased (4-256 mg/l) in the presence of new broad-spectrum plasmid mediated beta-lactamases (either TEM of SHV derivatives). Increased MICs (0.25-8 mg/l) were also observed for different isogenic mutants of Enterobacteriaceae, which either produced high levels of chromosome-encoded cephalosporinases, or had a permeability defect.

Anti-Bacterial Agents↗

[Evaluation of a new semi-automatic method of identifying enterobacteriaceae, M.I.S.-Enterobacteriaceae].

M.I.S.-Enterobacteriaceae is a new kit for identifying Enterobacteriaceae using a microplate consisting of 21 biochemical characters with automated reading and interpretation. The validity of this method was studied by the identification of 350 strains of enterobacteria belonging to 44 species and comparison with the classical method of identification in test-tubes. Results showed a diagnosis accuracy of 96 p. cent at the species level and 97.7 p. cent at the genus level. Diagnosis accuracy reached 100 p. cent for those bacterial species isolated routinely: Proteus, Providencia, Morganella, Klebsiella, Enterobacter. Accuracy was 97 p. cent for E. coli, 95 p. cent for Serratia marcescens and 94 p. cent for C. freundii. For several less frequently isolated species such as Salmonella, Hafnia, Shigella and Yersinia, diagnosis accuracy was 100 p. cent. This identification system for enterobacteria can however also be used for identification of Aeromonas genus and for Pseudomonas maltophilia and Acinetobacter baumannii non fermenting Gram-negative bacilli with oxidase negative reaction.

Bacteriological Techniques↗

Enterobacteriaceae in the jejunal microflora prevalence and relationship to biochemical and histological evaluations in healthy Colombian men.

When 23 healthy native Southwestern Colombian men were studied to determine the prevalence of Enterobacteriaceae in the jejunal microflora in a sample of thoroughly evaluated tropical inhabitants who were normal by physical examination, chest x-ray, and medical history, 14 of the 23 proved to be Enterobacteriaceae-positive, with counts of 10(3) to 10(9) per milliliter of jejunal aspirate. Thirteen had Escherichia coli, and the fourteenth had Klebsiella pneumoniae. Four had a second species of Enterobacteriaceae associated with E. coli: three were K. pneumoniae, and one was Proteus morganii. Laboratory studies routinely done on all subjects disclosed a total of 18 low biochemical values, 17 of which were associated with 12 of the 14 Enterobacteriaceae-positive subjects: six were low in serum cholesterol, four low in serum vitamin B12, four low in D-xylose excretion, and three low in creatinine coefficient; whereas, the Enterobacteriaceae-negative subjects had normal values for all biochemical tests except for serum vitamin B12 in one case. Nitrogen balance means were significantly different for the two groups: 3.39 g for the Enterobacteriaceae-positive subjects and 1.94 g for the Enterobacteriaceae-negative. No relationship was evident when the histology of the jejunal biopsies was compared with the microbiological or laboratory findings. When the 23 subjects were grouped into those (N = 19) with significant microbial recoveries of any type and those (N = 4) without, the data yielded no meaningful relationships.

Adolescent↗

Oligonucleotide probe for detecting Enterobacteriaceae by in situ hybridization.

AIMS: To develop oligonucleotide probes for visualizing bacteria belonging to Enterobacteriaceae. METHODS AND RESULTS: 24-mer oligonucleotide probe (probe D) was designed by comparison of 16S rDNA sequences of 35 species of Enterobacteriaceae, eight species of Vibrionaceae and six species of Pasteurellaceae. The sequence of the probe corresponding to the complementary sequence of a position 1251-1274 of Escherichia coli 16S rRNA was found to be a highly conserved region of 16S rDNA sequence in Enterobacteriaceae different from that of Vibrionaceae and Pasteurellaceae. The fluorescent dye-labelled probe was tested for the specificity by in situ hybridization and epifluorescence microscopy. Seventy-six out of 78 strains belonging to Enterobacteriaceae were visualized in an optimal hybridization condition. No bacterial strains belonging to Vibrionaceae (31 strains) and Gram-positive bacteria (three strains) were visualized. CONCLUSIONS: In situ hybridization using probe D allows the detection of bacterial cells belonging to Enterobacteriaceae without false positive reaction. SIGNIFICANCE AND IMPACT OF THE STUDY: In situ hybridization techniques using the probe D are potential tools for detecting Enterobacteriaceae in food and water samples.

DNA, Bacterial↗

Seven-hour fluorescence in situ hybridization technique for enumeration of Enterobacteriaceae in food and environmental water sample.

AIMS: A fluorescent in situ hybridization (FISH) technique using an Enterobacteriaceae-specific probe (probe D) to target 16S rRNA was improved in order to enumerate, within a single working day, Enterobacteriaceae present in food and environmental water samples. METHODS AND RESULTS: In order to minimize the time required for the FISH procedure, each step of FISH with probe D was re-evaluated using cultured Escherichia coli. Five minutes of ethanol treatment for cell fixation and hybridization were sufficient to visualize cultured E. coli, and FISH could be performed within 1 h. Because of the difficulties in detecting low levels of bacterial cells by FISH without cultivation, a FISH technique for detecting microcolonies on membrane filters was investigated to improve the bacterial detection limit. FISH with probe D following 6 h of cultivation to grow microcolonies on a 13 mm diameter membrane filter was performed, and whole Enterobacteriaceae microcolonies on the filter were then detected and enumerated by manual epifluorescence microscopic scanning at magnification of x100 in ca 5 min. The total time for FISH with probe D following cultivation (FISHFC) was reduced to within 7 h. FISHFC can be applied to enumerate cultivable Enterobacteriaceae in food (above 100 cells g-1) and environmental water samples (above 1 cell ml-1). CONCLUSIONS: Cultivable Enterobacteriaceae in food and water samples were enumerated accurately within 7 h using the FISHFC method. SIGNIFICANCE AND IMPACT OF THE STUDY: A FISHFC method capable of evaluating Enterobacteriaceae contamination in food and environmental water within a single working day was developed.

Colony Count, Microbial↗

Maternal risk factors for perinatal septicemia due to the Enterobacteriaceae.

Cases of perinatal septicemia due to the Enterobactericeae, which manifest in the first 24 hr of life were analyzed for the presence or absence of the maternal risk factors defined in the CDC group B streptococcus (GBS) risk-factor protocol. Microbiological data involving blood culture isolates were reviewed for the recovery of an Enterobacteriaceae from January 1975 through June 1995. Enterobacteriaceae perinatal septicemia was defined as the recovery an Enterobacteriaceae from one or more set of blood cultures in the first 24-36 hr of life in conjuncture with clinical evidence of neonatal stress in the first 24 hr. A case would also be considered of perinatal origin for cultures obtained up to 36 hr provided that evidence of clinical disease was present in the first 24 hr of life. Fifteen cases of perinatal septicemia due to the Enterobacteriaceae were analyzed. All but four shared one or more maternal risk factors. The maternal risk criteria established to avert early-onset GBS disease are commonly encountered in women destined to have newborns with perinatal septicemia due to the Enterobacteriaceae. Antibiotic selection for risk-driven protocols for GBS avoidance may need to be broadened to address the issue of coverage for penicillin-sensitive and penicillin-resistant Enterobacteriaceae.

Antibiotic Prophylaxis↗

[The presence of Candida spp and rods of Enterobacteriaceae family in the vaginal swab specimens].

The aim of the present work was to determine the presence and quantity of yeasts (Candida) and rods of Enterobacteriaceae family in the ontocenosis of vagina in women. Also, we aimed at finding which species of Enterobacteriaceae accompany the yeasts. Furthermore we aimed at determining the etiological factor (Candida yeasts or Enterobacteriaceae rods) causing identical clinical symptoms such as burn, itch, leucorrhea. Vagina secretion collected from 649 patients constituted the material for examination Candida. In 137 patients (which constituted 21.1%) Candida yeasts were found (group I). These patients were additionally divided into 2 subgroups (I a and I b). Group I a consisted of 92 patients (14.2%) who exhibited the presence of Candida without the accompanying rods of Enterobacteriaceae and group I b consisting of 45 patients (6.9%), in whom Candida was found together with the rods. 171 patients (26.3%) were classified to group II, who had only Enterobacteriaceae rods in the vagina. Among Candida yeasts, the most common species were C. albicans, C. krusei, C. tropicalis. The most common rods of Enterobacteriaceae family were E. coli and P. mirabilis. The bacterium which most frequently accompanied the yeasts was E. coli. Considering the fact that patients in group I and patients in group II reported similar problems, the role of the etiological factor in the vaginal inflammatory processes should be analyzed.

Candida↗

[Current aspects of the fecal flora of the newborn without antibiotherapy during the first 7 days of life: Enterobacteriaceae, enterococci, staphylococci].

Last years, il became obvious that the colonization pattern described in 1976-1978 was no more valid: early colonization by Enterobacteriaceae at the 2-3 rd day of life in all newborns, with constant presence of antibioresistant strainseven in non treated newborns. To establish the new pattern of colonization, the same quantitative method of dilution and culture on selective media was used daily from day 1 to day 7 (5 days only for M). The number of Enterobacteriaceae, enterococci and staphylococci was determined in the stools of 10 newborns in the Maternity unit (= M) (term 40 weeks +/- 1, birth weight 3,356 g +/- 383), 10 in the Premature nursery (= P) (term 34.9 weeks +/- 1, birth weight 2,457 g +/- 676), and 14 in the Neonatal intensive care unit (= R) (term 35.2 weeks +/- 3.8, birth weight 2,457 g +/- 763). The results establish that colonization by Enterobacteriaceae is no more constant at D3. It could be demonstrated only in 8/10 M, 1/10 P, and 6/14 R (statistically different - p < 0.01 - between M and P). At D5, 9/10 M, 5/10 P, 10/14 R, and at D7, 6/10 P and 10/14 R were colonized. Resistant Enterobacteriaceae (Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae) could be found in only 3/10 M, 4/10 P and 6/14 R. Enterococci could be found in 1 newborn M, 2 P and 7 newborns R. Staphylococci appeared earlier: all newborns M, P and R were colonized at D2, 4 and 5 respectively. These bacteria were coagulase negative, associated with Staphylococcus aureus in 3 P. Our hypothesis is that late colonization with Enterobacteriaceae and enterococci is due to the improvement of hygiene procedures and due to the decontaminating effect of antibiotics in other treated newborns (Enterobacteriaceae by 3 rd generation cephalosporin and enterococci by pharyngeal vancomycin).

Anti-Bacterial Agents↗

Inhibition of Salmonella typhimurium attachment to chicken cecal mucus by intestinal isolates of Enterobacteriaceae and lactobacilli.

The ability of selected strains of Enterobacteriaceae or lactobacilli isolated from the intestines of adult chickens to inhibit in vitro attachment of Salmonella typhimurium 3333/O to cecal mucus in the presence or absence of D-mannose was determined. Attachment in the absence of mannose was reduced by prior exposure of mucus to cultures of two isolates of Enterobacteriaceae, an Escherichia coli and a Hafnia alvei strain, but not to a third isolate, an Enterobacter agglomerans strain. Attachment of S. typhimurium was not inhibited when mannose was present in the blocking or attachment step. Formation of fimbriae by the two inhibitory Enterobacteriaceae strains and the S. typhimurium strain, as indicated by titers of mannose-sensitive hemagglutination of guinea pig erythrocytes was optimal in Z biphasic medium (consisting of tryptone, yeast extract, dextrose, and NaCl) incubated anaerobically at 42 C. Fimbriae of each of three strains prepared from these cultures also inhibited attachment. These are characteristics consistent with attachment and inhibition of attachment mediated by a mannose-sensitive adhesin associated with type 1 fimbriae on bacterial cells of Enterobacteriaceae strains. Attachment in the presence of mannose was significantly reduced by prior exposure of mucus to cultures of a Lactobacillus salivarius strain and a Lactobacillus delbrueckii delbrueckii strain but not to a strain of Lactobacillus for which the species had not been determined. Washed cells or spent culture supernatant fluid from brain-heart infusion broth, Z broth, or Z biphasic cultures of the inhibitory strains of lactobacilli incubated at 37 or 42 C inhibited this form of attachment. Of 27 intestinal isolates of Enterobacteriaceae and 21 of lactobacilli, the lactobacilli strains were generally more hydrophobic than the Enterobacteriaceae as determined by adherence to hexadecane. The lactobacilli isolates did not agglutinate guinea pig erythrocytes. The data suggest more than one mechanism for mediating attachment of inhibitory bacterial strains and for subsequent attachment of S. typhimurium.

Adhesins, Bacterial↗