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The gram-negative fecal floras from 953 infants were studied upon discharge of the infants from 22 neonatal wards. More than 600 distinct phenotypes of Escherichia coli, Klebsiella spp., and Enterobacter spp. were distinguished by high-resolution biotyping. The colonization patterns observed showed considerable local and temporal variation. The major (M) strains (phenotypes), which colonized more than 10% and up to 78% of the infants in a ward (median, 23%), were Klebsiella oxytoca (15 strains), E. coli (4 strains), Klebsiella pneumoniae (1 strain), and Enterobacter cloacae (1 strain). Resistance to beta-lactam antibiotics was more pronounced among M strains than among strains of enteric bacteria colonizing few or single infants only. Local antibiotic policy influenced the colonization patterns. Despite the fact that M strains of Klebsiella spp. were usually resistant to ampicillin as well as to cephalexin and cefuroxime, their local dissemination was associated with the use of ampicillin with or without gentamicin but not with the use of cefuroxime. It thus appeared that in the neonatal setting, ampicillin posed a greater risk of local spread of certain drug-resistant bacterial clones than a newer cephalosporin, such as cefuroxime.
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Forty-one stock organisms and 581 fresh clinical isolates were used in comparing the Minitek system to conventional tubed media to determine if this system is feasible and accurate for a high-volume clinical microbiology laboratory. In addition to comparison with tubed media, the following parameters were tested: (i) reproducibility of disks, (ii) the effect of variation in inoculum size, (iii) the effect of the age of culture, and (iv) the effect of predispensing disks. A total of 5,947 disks were compared with tube reactions; 95.9% agreed on the initial reading. After repeating questionable tube and disk reactions, this agreement rose to 98.2%. Using the disk reactions and the identification schema currently in use with tubed media, 94.9% of the Minitek final identifications agreed with those of tubed media; 1.5% gave differing identifications and 3.6% were inconclusive. This system is accurate, inexpensive, flexible, and convenient.
Susceptibility to piperacillin was similar to that to piperacillin-tazobactam (<1% difference) for 6,938 isolates of Enterobacter aerogenes and 13,954 isolates of Enterobacter cloacae tested using a Vitek system; for the same species, in contrast, susceptibility rates to piperacillin-tazobactam were 5.9 to 13.9% higher than to piperacillin using disk diffusion, MicroScan, and Vitek 2 testing. Unprecedented phenotypes (piperacillin susceptible and piperacillin-tazobactam intermediate; piperacillin intermediate and piperacillin-tazobactam resistant; piperacillin susceptible and piperacillin-tazobactam resistant) accounted for 6.1% of the results for E. aerogenes isolates and 6.0% of the results for E. cloacae isolates tested with the Vitek system.
With a modification of Taylor's decarboxylation broth, histidine decarboxylase was detected in Enterobacter aerogenes, Morganella morganii, Raoultella ornithinolytica, and some strains of Citrobacter youngae and Raoultella planticola. This method provides a useful confirmatory test for identification of E. aerogenes strains.
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Although plasmid-mediated AmpC beta-lactamases were first reported in the late 1980s, many infectious disease personnel remain unaware of their clinical importance. These enzymes are typically produced by isolates of Escherichia coli, Klebsiella spp., Proteus mirabilis, and Salmonella spp. and are associated with multiple antibiotic resistance that leaves few therapeutic options. Plasmid-mediated AmpC beta-lactamases have been associated with false in vitro susceptibility to cephalosporins. Many laboratories do not test for this resistance mechanism because current tests are inconvenient, subjective, lack sensitivity and/or specificity, or require reagents that are not readily available. In this study a new test, the AmpC disk test, based on filter paper disks impregnated with EDTA, was found to be a highly sensitive, specific, and convenient means of detection of plasmid-mediated AmpC beta-lactamases in organisms lacking a chromosomally mediated AmpC beta-lactamase. Using cefoxitin insusceptibility as a screen, the test accurately distinguished AmpC and extended-spectrum beta-lactamase production and differentiated AmpCs from non-beta-lactamase mechanisms of cefoxitin insusceptibility, such as reduced outer membrane permeability. The test is a potentially useful diagnostic tool. It can provide important infection control information and help to ensure that infected patients receive appropriate antibiotic therapy.
A modified Voges-Proskauer test is described which distinguishes within 4 to 8 hours between organisms that can produce acetyl-methyl-carbinol (acetoin) from glucose fermentation and those that cannot.
In Escherichia coli K-12, the receptor for phage lambda is an outer membrane protein which inactivates the phage in vitro. Lambda receptor activity was found in extracts from all wild strains of E. coli tested, although most of them fail to support growth of the phage. In some cases this failure is due to a masking of the receptor in vivo, the bacteria being unable to adsorb the phage or to react with antireceptor antibodies. In other cases, adsorption does occur, and the nature of the block in phage growth was not investigated. Most Mal+ strains of Shigella have lambda receptor, whereas most Mal- strains do not have it. Synthesis of the lambda receptor in Shigella is thus presumably controlled by the positive regulator gene of the maltose regulon as is the case in E. coli K-12. Phage lambda adsorbs on many Mal+ strains of Shigella and even yields plaques on some of them, although at a low frequency. No lambda receptor activity could be found in extracts of several strains of Salmonella and Levinea.
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Two isolates of Proteus mirabilis and four of Esch. coli which would not grow on commonly used commercial sensitivity test media are reported. These organisms appear to be resistant to co-trimoxazole, are exacting towards thymidine, and five of six were isolated from patients who had been treated with co-trimoxazole. A method for the detection of these strains is given and their significance in the laboratory and clinical practice discussed.
Two hundred and twenty-two organisms were identified by conventional methods and by the Modified R/B Enteric Differential System. Two hundred and eighteeen (98-2%) of these organisms were correctly identified by the R/B system. The results of individual tests were also compared. The convenience and general ease of handling of the R/B system are discussed.
Six conventional biochemical tests were combined to produce an identification scheme. These tests included decarboxylation of lysine and ornithine, fermentation of glucose and cellobiose, indole production and urease production. Three hundred antibiotic resistant coliforms from urine specimens were tested by this scheme and also by the API 20E for comparison. Two hundred and seventy nine (93%) of organisms were correctly identified using the six tests, 17 (5.7%) were referred for further study, and four (1.3%) were misidentified. It is concluded that this combination of tests provides an inexpensive, accurate, and rapid tool for identification.
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Xenorhabdus nematophilus mutants that took longer to kill insects than did the wild type were used to determine the relationship of the physicochemical properties and outer membrane composition to bacterial interaction with the antibacterial systems of Galleria mellonella larvae and to bacterial virulence. Insect serum slowed the growth of the wild-type and mutant bacteria. This was attributed to increased spheroplast formation for the mutants. Spheroplast formation was associated with an increased sensitivity to insect lysozyme and a reduction in overall bacterial cationic charge. Increasing bacterial hydrophobicity was correlated with both increased bacterial attachment to the insect's haemocytes and the accelerated removal of the bacteria from the haemolymph. Attachment of the mutants to the insect haemocytes also increased as the bacterial lipopolysaccharide content increased, the level of prophenoloxidase activation increased, and cationic charge declined. Bacterial emergence into the haemolymph occurred in parallel with haemocyte damage but neither the total lipopolysaccharide levels in the bacteria nor the rate of bacterial emergence were associated with virulence. The rate of lipopolysaccharide release into the haemolymph influenced the rate of haemocyte damage. The contribution of outer membrane proteins to lipopolysaccharide release, bacterial adhesion to haemocytes, and virulence is discussed. Virulence reflects bacterial tolerance to the host's antibacterial defences, favouring an increase in bacteria and toxic lipopolysaccharides.
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