Experimental model of oral antityphoid vaccination with live streptomycin-dependent Salmonella typhimurium in C57BL/6 mice.
(57/1407)
The present experimental model offered the opportunity to study particular aspects concerning the avirulence stability of live streptomycin-dependent (Sm D) Salmonella vaccines, under conditions resembling human enteric fever. The results seem to indicate that, in practice, the risk of reversion to a virulent form during oral antityphoid vaccination with Sm D strains remains slight even after the interruption of concomitant streptomycin administration. (+info)
Evaluation of Etest for susceptibility testing of Mycobacterium tuberculosis.
(58/1407)
The Etest method for susceptibility testing of Mycobacterium tuberculosis was compared to the agar proportion method using four first-line agents and two fluoroquinolones. Catergorical agreement between the methods was 100% for rifampin, ethambutol, streptomycin, and ofloxacin and 98% for isoniazid. Results were obtained in 6 to 10 days by Etest. The Etest method is suitable for testing the agents evaluated against M. tuberculosis. (+info)
Antibiotic resistance and survival of E coli in the alimentary tract.
(59/1407)
Some antibiotics tend to select for R-factor-carrying Escherichia coli in the human gut, with complex long-term consequences. Some resistant strains disappear rapidly when treatment ends, while others persist for months in the absence of obvious antibiotic selection pressure, and the performance of individual resistant strains seems to depend more on the nature of the strain than on the plasmid carried. R plasmids are relatively rare in those E coli that colonize well in the gut and resistant bacteria therefore tend to disappear when treatment ends, but this situation could change dramatically if R plasmids became prevalent among those strains of E coli that colonize effectively. (+info)
Role of Pseudomonas aeruginosa PhoP-phoQ in resistance to antimicrobial cationic peptides and aminoglycosides.
(60/1407)
Resistance to the polycationic antibiotic polymyxin B and expression of the outer-membrane protein OprH in the opportunistic pathogen Pseudomonas aeruginosa both involve the PhoP-PhoQ two-component regulatory system. The genes for this system form an operon with oprH, oprH-phoP-phoQ, that responds to Mg(2+) starvation and PhoP levels. In this study, the Mg(2+)-regulated promoter for this operon was mapped upstream of oprH by primer-extension experiments. An oprH::xylE-Gm(R) mutant H855 was constructed and measurement of the catechol 2,3-dioxygenase activity expressed from this transcriptional fusion provided evidence for a second, weak promoter for phoP-phoQ. Wild-type P. aeruginosa PAO1 strain H103 was found to exhibit Mg(2+)-regulated resistance to the alpha-helical antimicrobial cationic peptide CP28 in addition to its previously characterized resistance to polymyxin B. Resistance to this peptide was unchanged in the OprH-null mutant H855 and a PhoP-null mutant H851. In contrast, PhoQ-null mutant H854 demonstrated constitutive CP28 resistance. Northern blot analysis revealed constitutive expression of phoP in this strain, implicating PhoP-PhoQ in the resistance of P. aeruginosa to cationic peptides. Furthermore, all three null-mutant strains demonstrated increased resistance to the aminoglycoside antibiotics streptomycin, kanamycin and amikacin. Two additional mutant strains, H895 and H896, were constructed that carried unmarked deletions in oprH and were found to exhibit aminoglycoside susceptibility equivalent to that of the wild-type. This result provided definitive evidence that OprH is not involved in P. aeruginosa aminoglycoside resistance and that the changes in resistance in strain H855 and a previously reported oprH mutant were due to polar effects on phoP-phoQ rather than loss of OprH expression. A role for PhoP-PhoQ in resistance to aminoglycosides is envisaged that is distinct from that in resistance to cationic peptides and polymyxin B. (+info)
Necrosis of lung epithelial cells during infection with Mycobacterium tuberculosis is preceded by cell permeation.
(61/1407)
Mycobacterium tuberculosis establishes infection, progresses towards disease, and is transmitted from the alveolus of the lung. However, the role of the alveolar epithelium in any of these pathogenic processes of tuberculosis is unclear. In this study, lung epithelial cells (A549) were used as a model in which to examine cytotoxicity during infection with either virulent or avirulent mycobacteria in order to further establish the role of the lung epithelium during tuberculosis. Infection of A549 cells with M. tuberculosis strains Erdman and CDC1551 demonstrated significant cell monolayer clearing, whereas infection with either Mycobacterium bovis BCG or Mycobacterium smegmatis LR222 did not. Clearing of M. tuberculosis-infected A549 cells correlated to necrosis, not apoptosis. Treatment of M. tuberculosis-infected A549 cells with streptomycin, but not cycloheximide, demonstrated a significant reduction in the necrosis of A549 cell monolayers. This mycobacterium-induced A549 necrosis did not correlate to higher levels of intracellular or extracellular growth by the mycobacteria during infection. Staining of infected cells with propidium iodide demonstrated that M. tuberculosis induced increased permeation of A549 cell membranes within 24 h postinfection. Quantitation of lactate dehydrogenase (LDH) release from infected cells further demonstrated that cell permeation was specific to M. tuberculosis infection and correlated to A549 cellular necrosis. Inactivated M. tuberculosis or its subcellular fractions did not result in A549 necrosis or LDH release. These studies demonstrate that lung epithelial cell cytotoxicity is specific to infection by virulent mycobacteria and is caused by cellular necrosis. This necrosis is not a direct correlate of mycobacterial growth or of the expression of host cell factors, but is preceded by permeation of the A549 cell membrane and requires infection with live bacilli. (+info)
Genetic loci responsible for incompatibility on a co-integrate plasmid, R100-1.
(62/1407)
An R plasmid, R100-1, was mapped previously (Yoshikawa, 1974) by transduction from an integratively suppressed Hfr strain to a recipient with a mutation in gene dnaA. By this method various types of transductants of plasmid R100-1 that exist autonomously or in the integrated state were obtained. Seventy-one such transductants were used in the present study to map gene inc, which is responsible for incompatibility. The results obtained can be explained by either of the following: (i) R100-1 has only a single gene or gene cluster (inc) despite previous work suggesting that this plasmid is a co-integrate of two replicons; (ii) R100-1 possesses more than one inc locus located between the repA and tra loci. (+info)
Compatibility of pTM89, a new F-like R factor, and of derivative plasmids.
(63/1407)
pTM89, an fi+ R factor that controls the production of repressed F-type pili, is incompatible with plasmids belonging to the FII and P groups. The results of P1 transduction show that all of the resistance markers of pTM89 are part of a single replicon, which also includes RTF. When the compatibility of different derivative plasmids was investigated, it was found that they fall into two classes. Those of the first class have lost the compatibility of pTM89 for the P group but are still incompatible with FII group, whereas those of the second class are compatible with plasmids of both groups. Plasmids of the latter class that are also compatible with each other and, therefore, apparently lack any determinant for compatibility are genetically stable and self-transmissible. It appears, therefore, that compatibility between plasmids cannot be explained by the hypothesis of competition for a maintenance site. (+info)
Denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA amplicons to monitor changes in fecal bacterial populations of weaning pigs after introduction of Lactobacillus reuteri strain MM53.
(64/1407)
The diversity and stability of the fecal bacterial microbiota in weaning pigs was studied after introduction of an exogenous Lactobacillus reuteri strain, MM53, using a combination of cultivation and techniques based on genes encoding 16S rRNA (16S rDNA). Piglets (n = 9) were assigned to three treatment groups (control, daily dosed, and 4th-day dosed), and fresh fecal samples were collected daily. Dosed animals received 2.5 x 10(10) CFU of antibiotic-resistant L. reuteri MM53 daily or every 4th day. Mean Lactobacillus counts for the three groups ranged from 1 x 10(9) to 4 x 10(9) CFU/g of feces. Enumeration of strain L. reuteri MM53 on MRS agar (Difco) plates containing streptomycin and rifampin showed that the introduced strain fluctuated between 8 x 10(3) and 5 x 10(6) CFU/g of feces in the two dosed groups. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA fragments, with primers specific for variable regions 1 and 3 (V1 and V3), was used to profile complexity of fecal bacterial populations. Analysis of DGGE banding profiles indicated that each individual maintained a unique fecal bacterial population that was stable over time, suggesting a strong host influence. In addition, individual DGGE patterns could be separated into distinct time-dependent clusters. Primers designed specifically to restrict DGGE analysis to a select group of lactobacilli allowed examination of interspecies relationships and abundance. Based on relative band migration distance and sequence determination, L. reuteri was distinguishable within the V1 region 16S rDNA gene patterns. Daily fluctuations in specific bands within these profiles were observed, which revealed an antagonistic relationship between L. reuteri MM53 (band V1-3) and another indigenous Lactobacillus assemblage (band V1-6). (+info)