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Effect of subinhibitory concentrations of clindamycin and trospectomycin on the adherence of Staphylococcus epidermidis in an in vitro model of vascular catheter colonization.

Septicemia, often due to Staphylococcus epidermidis, is a life-threatening complication associated with indwelling vascular catheters. An important factor in the development of such infections is glycocalix, or slime. An in vitro model that mimics intravenous delivery systems in humans was developed. It consisted of a modified Robbins device containing slices of silicone catheters in the removable ports, through which S. epidermidis diluted in 5% dextrose-normal saline with 10% heat-inactivated normal human serum was run, with and without clindamycin and trospectomycin. S. epidermidis was recovered from all catheters in the absence of antibiotics; no growth was detected with antibiotics. Scanning electron microscopy demonstrated significant reduction in glycocalix and no visible organisms with all concentrations except 0.5 micrograms/ml trospectomycin and 1 microgram/ml clindamycin; for those, a moderate amount of glycocalix and a few bacteria were seen. Thus, subinhibitory levels of trospectomycin and clindamycin may have a role in the prevention of microbial adherence to vascular catheters.

Anti-Bacterial Agents↗

Transfer of pheromone-inducible plasmids between Enterococcus faecalis in the Syrian hamster gastrointestinal tract.

Pheromone-responsive plasmids are common to Enterococcus faecalis, transfer at high frequency in vitro, and carry cytolysin and other gene products implicated in the pathogenesis of enterococcal infection. A Syrian hamster model of enterococcal intestinal overgrowth was used to test for transfer of three isogenic plasmids differing in conjugative and cytolytic phenotypes. Transconjugants were found in 8 (44%) of 18 and 6 (35%) of 17 hamsters given donor strains containing cytolytic (pAM714) and noncytolytic (pAM771) pheromone-responsive plasmids. Of the 14 hamsters from which transconjugants were isolated from stool, 9 (64%) had transconjugants 1 day after donor strain inoculation. The frequency of transfer (mean +/- SD) for pAM714 and pAM771 was 1.4 +/- 2.2 x 10(-1) and 2.9 +/- 4.2 x 10(-2) transconjugants/donor, respectively (P > .20). Transconjugants were not recovered from hamsters receiving a cytolytic, nonconjugative plasmid (pAM930; transfer frequency < 2 x 10(-5) transconjugants/donor). Pheromone-responsive plasmid transfer between E. faecalis strains occurs at high frequency in the gastrointestinal tract of hamsters and may be one means by which enterococcal resistance and virulence factors disseminate.

Amino Acid Sequence↗

Conjugal transfer of penicillin resistance plasmids to proline-citrulline-uracil dependent strains of Neisseria gonorrhoeae.

Proline-, citrulline-, uracil (PCU)-dependent Neisseria gonorrhoeae represented only 2% of all Neisseria gonorrhoeae isolated in Greece during 1984-1987. We investigated whether these strains could be recipients of gonococcal plasmids. We carried out bacterial conjugation experiments and obtained the transfer of the African (3.2 Mdal) and of the Asian (4.5 Mdal) penicillin resistance plasmid from clinical penicillinase producing N. gonorrhoeae (PPNG) isolates to PCU gonococcal strains. The transfer frequency was very low, which suggests an explanation of the fact that the wild type PCU strains do not harbour either of these two plasmids. The conjugative plasmid of 24.5 Mdal was not transferred to any of the transconjugants carrying either the African or the Asian type plasmids.

Cephalosporins↗

In-vitro activities of trospectomycin, cefpodoxime, and second-generation cephalosporins against Haemophilus influenzae type b.

The in-vitro activities of trospectomycin, cefpodoxime, cefamandole, cefonicid, and cefuroxime against beta-lactamase-negative and -positive invasive clinical isolates of Haemophilus influenzae type b were determined by the agar dilution method. Trospectomycin and cefpodoxime inhibited 90% of the strains at concentrations of 5 and 0.06 mg/l, respectively, and no differences between the susceptibilities of the beta-lactamase-negative and -positive strains were noted. The activity of cefpodoxime was minimally affected by increased inoculum size, but significant inoculum effects were noted with cefamandole, cefonicid, and cefuroxime with beta-lactamase positive strains.

Anti-Bacterial Agents↗

Susceptibility of 539 gram-positive and gram-negative anaerobes to new agents, including RP59500, biapenem, trospectomycin and piperacillin/tazobactam.

Susceptibilities of 539 Gram-positive and Gram-negative anaerobes were tested by agar dilution against 15 new and existing antimicrobial agents. Organisms included 218 Bacteroides fragilis group strains, 15 non-fragilis group Bacteroides, 130 Porphyromonas/Prevotella, 49 fusobacteria, 50 peptostreptococci, 53 clostridia and 24 Gram-positive non-sporeforming bacilli. Of 412 Gram-negative bacilli, 89% were beta-lactamase-positive, while only two of the Gram-positive strains (both clostridia) produced this enzyme. Using established and preliminary breakpoints, all strains were susceptible to biapenem and imipenem (MIC90s 1 mg/L) and chloramphenicol (MIC90 8 mg/L). Only one of all the strains tested (a Cl. innocuum) was resistant (MIC > 4 mg/L) to RP59500; the latter had MIC90 2 mg/L, while 98% of strains were susceptible to trospectomycin (MIC90 16 mg/L). Ninety-nine per cent of strains were susceptible to piperacillin/tazobactam (MIC90 8 mg/L) compared to 86% to piperacillin (MIC90 > 64 mg/L). Corresponding data for ticarcillin/clavulanate versus ticarcillin were 97% susceptible (MIC90 8 mg/L) compared to 83% (MIC90 > 64 mg/L). Enhancement of the beta-lactam by the inhibitors was only seen in beta-lactamase-producing strains. Amoxycillin and cefoperazone were less often active (36% susceptible, MIC90 > 256 mg/L and 66% susceptible, MIC90 64 mg/L, respectively). Cefoxitin had greater activity than cefotetan (90% susceptible, MIC90 32 mg/L, compared to 72% susceptible, MIC90 > 64 mg/L). Metronidazole was active against 94% of strains (MIC90 4 mg/L). All metronidazole-resistant strains were Gram-positive (75% of non-sporeforming bacilli, 9% of clostridia and 6% of peptostreptococci). Ninety per cent of strains were susceptible to clindamycin (MIC90 4 mg/L).

Anti-Bacterial Agents↗

The spc ribosomal protein operon of Escherichia coli: sequence and cotranscription of the ribosomal protein genes and a protein export gene.

The genes encoding the 52 ribosomal proteins (r-proteins) of Escherichia coli are organized into approximately 19 operons scattered throughout the chromosome. One of these, the spc operon, contains the genes for ten ribosomal proteins: L14, L24, L5, S14, S8, L6, L18, S5, L30 and L15 (rp1N, rp1X, rp1E, rpsN, rpsH, rp1F, rp1R, rpsE, rpmD, and rp1O). We now report the entire 5.9 kb nucleotide sequence of the spc operon. DNA sequence analysis has confirmed the genetic organization and refined the amino acid sequence of the ten r-proteins in this operon. It has also revealed the presence of two open reading frames past the last known gene (L15) of the spc operon. One of these corresponds to a gene (pr1A or secY) which recently has been shown by others to be involved in protein export. In addition, S1 mapping experiments indicate that a significant proportion of transcription initiated from the spc operon continues not only into the two putative genes, but also without termination into the downstream alpha r-protein operon.

Bacterial Proteins↗

The region of the IncN plasmid R46 coding for resistance to beta-lactam antibiotics, streptomycin/spectinomycin and sulphonamides is closely related to antibiotic resistance segments found in IncW plasmids and in Tn21-like transposons.

The nucleotide sequence of a 2.5 kb segment of the pKM101 (R46) genome has been determined. The 1.3 kb from a BamHI site at 153 to base 1440 differs by only 2 bases from a part of the published sequence of the aadB (gentamicin resistance) gene region including the coding region for the N-terminal 70 amino acids of the predicted aadB product. The same sequence has been found 5'-to the dhfrII gene of R388 and to the aadA gene of Tn21 (R538-1). Three open reading frames are located in this region, two on the same strand as the resistance genes and one on the complementary strand. The latter predicts a polypeptide of 337 amino acids, whose N-terminal segment is 40% homologous to the predicted product of an open reading frame of 179 amino acids located next to the dhfrI gene of Tn7. The oxa2 (oxacillin resistance) gene predicts a long polypeptide commencing with (the N-terminal) 70 amino acids of the aadB product. A similar arrangement is found in the aadA gene of R538-1. The N-terminal segment of an aadA gene is located 3'- to oxa2, separated by 36 bases. Sequences surrounding the BamHI site are identical to sequences 5'- to the tnpM gene of Tn21 and homology ceases where homology between Tn21 and Tn501 commences. The possibility that this antibiotic resistance segment is a discrete mobile DNA element is discussed.

Amino Acid Sequence↗

Site-selective inhibition of plastid RNA editing by heat shock and antibiotics: a role for plastid translation in RNA editing.

RNA editing in higher plant plastids changes single cytidine residues to uridine through an unknown mechanism. In order to investigate the relation of editing to physiological processes and to other steps in plastid gene expression, we have tested the sensitivity of chloroplast RNA editing to heat shock and antibiotics. We show that heat shock conditions as well as treatment of plants with prokaryotic translational inhibitors can inhibit plastid RNA editing. Surprisingly, this inhibitory effect is confined to a limited number of plastid editing sites suggesting that some site-specific factor(s) but none of the general components of the plastid RNA editing machinery are compromised. Contrary to previous expectations, our results provide evidence for a role of plastid translation in RNA editing.

Anti-Bacterial Agents↗