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| J Bacteriol, 1994 Aug, 176(16), 5140 - 4 Characterization of aarA, a pleiotrophic negative regulator of the 2'-N-acetyltransferase in Providencia stuartii; Rather PN et al.; We have utilized transposon mutagenesis to obtain insertional mutations in Providencia stuartii that activate the chromosomal aac(2')-la gene . Two closely linked mini-Tn5Cm insertions were obtained in a locus designated aarA, and a single insertion was obtained in a separate locus, aarC . Nucleotide sequence analysis, complementation studies, and localization of the sites of mini-Tn5Cm insertion have allowed the identification of the aarA coding region . The deduced AarA protein had a molecular mass of 31,086 kDa and displayed characteristics of an integral membrane protein . A strain deleted for the aarA gene by allelic exchange showed at least a fourfold increase in the accumulation of aac(2')-la mRNA and an eightfold increase in aminoglycoside resistance . Mutations in aarA were pleiotrophic and also resulted in loss of pigmentation and a deficiency in cell separation during division. J Bacteriol, 1993 Oct, 175(20), 6492 - 8 Characterization and transcriptional regulation of the 2'-N-acetyltransferase gene from Providencia stuartii; Rather PN et al.; We have cloned the chromosomally encoded 2'-N-acetyltransferase gene {aac(2')-Ia} from Providencia stuartii . DNA sequence analysis of the cloned insert identified a single open reading frame, which is capable of encoding a protein with a predicted molecular mass of 20,073 Da . The deduced AAC(2')-Ia protein showed no significant homology to other proteins, including all of the AAC(3) and AAC(6') proteins . Primer extension analysis was used to identify the aac(2')-Ia promoter, which contained an unusual sequence (CTTTTT) at the -35 region . Expression of the aac(2')-Ia gene occurs at low levels in wild-type P . stuartii strains; therefore, they are aminoglycoside susceptible . We have isolated mutants with high-level AAC(2')-Ia expression at a frequency of 4.8 x 10(-6) . Detailed analysis of one mutant demonstrated a 12.2-fold increase in the accumulation of aac(2')-Ia mRNA . In addition, the levels of beta-galactosidase expression from a plasmid-encoded aac(2')-lacZ transcriptional fusion were increased 11.5-fold in this mutant relative to those in an isogenic wild-type strain . These results suggested that a trans-acting factor, designated aar (for aminoglycoside acetyltransferase regulator), controls AAC(2')-Ia expression in P . stuartii. J Pathol, 1993 Sep, 171(1), 67 - 71 Electron microscopic study of the attachment and penetration of rabbit intestinal epithelium by Providencia alcalifaciens; Mathan MM et al.; Ilea of adult rabbits with removable ileal ties (RITARD model) that developed diarrhoea and ileal loops of adult rabbits after inoculation with Providencia alcalifaciens, isolated from two patients with diarrhoea, were studied by transmission electron microscopy for enterocyte-bacterial interactions . Two modes of entry of bacteria into epithelial cells were seen: one directly by endocytosis associated with polymerization of cytoskeletal components and the other by disruption of tight junctions with entry into and proliferation in intercellular spaces. New Microbiol, 1993 Apr, 16(2), 149 - 64 Rapid access to pharmacokinetics data and correlation between antimicrobial susceptibility results and drug tissue distribution using a personal computer; Cornaglia G et al.; MYMIC is a computer-aided system capable of integrating antibiotic susceptibility data with the concentrations the drugs reach in various body tissues and fluids by calculating site concentration/minimal inhibitory concentration quotients . The program can be run on any low-cost personal computer operating under MS-DOS, provided it is equipped with a hard-disk drive and with a minimum of 512 kilobytes of random access memory . The use of the program does not require any knowledge of computer languages . The antibiotic susceptibility data can be entered either as minimal inhibitory concentrations or as inhibitory zone diameters; in the latter case, minimal inhibitory concentrations are automatically calculated via regression formulas . The concentrations obtained by 90 antibiotics in 51 different human tissues and fluids are recorded in a data base of over 1,000 records, obtained from roughly 700 original papers . A MYMIC sample session was simulated by mimicking infections of three different body districts (namely bone, prostate, and sputum) caused by Pseudomonas aeruginosa, Escherichia coli or Providencia stuartii. Nucleic Acids Res, 1992 Dec 25, 20(24), 6517 - 23 BsuBI--an isospecific restriction and modification system of PstI: characterization of the BsuBI genes and enzymes; Xu GL et al.; The enzymes of the Bacillus subtilis BsuBI restriction/modification (R/M) system recognize the target sequence 5'CTGCAG . The genes of the BsuBI R/M system have been cloned and sequenced and their products have been characterized following overexpression and purification . The gene of the BsuBI DNA methyltransferase (M.BsuBI) consists of 1503 bp, encoding a protein of 501 amino acids with a calculated M(r) of 57.2 kD . The gene of the restriction endonuclease (R.BsuBI), comprising 948 bp, codes for a protein of 316 amino acids with a predicted M(r) of 36.2 kD . M.BsuBI modifies the adenine (A) residue of the BsuBI target site, thus representing the first A-N6-DNA methyltransferase identified in B . subtilis . Like R.PstI, R.BsuBI cleaves between the A residue and the 3' terminal G of the target site . Both enzymes of the BsuBI R/M system are, therefore, functionally identical with those of the PstI R/M system, encoded by the Gram negative species Providencia stuartii . This functional equivalence coincides with a pronounced similarity of the BsuBI/PstI DNA methyltransferases (41% amino acid identity) and restriction endonucleases (46% amino acid identity) . Since the genes are also very similar (58% nucleotide identity), the BsuBI and PstI R/M systems apparently have a common evolutionary origin . In spite of the sequence conservation the gene organization is strikingly different in the two R/M systems . While the genes of the PstI R/M system are separated and transcribed divergently, the genes of the BsuBI R/M system are transcribed in the same direction, with the 3' end of the M gene overlapping the 5' end of the R gene by 17 bp. Res Microbiol, 1991 Nov-Dec, 142(9), 965 - 9 Correlation between DNA polymorphism and enzyme polymorphism argues in favour of the delineation of two species within Providencia alcalifaciens; Picard B et al.; Ribosomal DNA (rDNA) polymorphism was compared to enzyme polymorphism and DNA/DNA hybridization data for the intraspecies differentiation of Providencia alcalifaciens . DNA from 27 strains previously classified into two zymotypes A1 and A2 and discriminated by two levels of DNA/DNA hybridization (delta Tm values of 0 to 1 degree C and 6 to 10 degrees C, respectively) were analysed by Southern blotting for rDNA polymorphism . The ribotypes fell into two ribogroups A1 and A2, which correlated with the corresponding zymotypes . This correlation argues for the proposed creation of two species within P . alcalifaciens. J Econ Entomol, 1991 Feb, 84(1), 185 - 90 Oviposition by screwworm flies (Diptera: Calliphoridae) on contact with host fluids; Hammack L; Factor affecting oviposition by screwworm flies, Cochliomyia hominivorax (Coquerel), contacting different host fluids were examined in a laboratory bioassay . Fresh bovine blood, which does not release the attractive odors involved in host finding, nevertheless stimulated as many or more females to oviposit than did the other fluids tested . These other fluids included attractive fluid from screwworm-infected wounds (a favored oviposition site in nature) and cultures of Providencia rettgeri (a bacterium implicated in attractant production) . Oviposition did not vary with batch of fresh blood or frozen storage, making blood a useful standard for comparing oviposition rates among studies . Oviposition did vary with the substrate to which the blood was applied, suggesting that an interaction between tactile and chemical stimuli is important for host recognition . Both insemination and darkness during bioassay increased oviposition rates, but the magnitude of these effects was small compared with that due to substrate . Age had no effect for at least 1 wk after females became gravid. J Infect Dis, 1990 Dec, 162(6), 1354 - 9 Bacterial infections in patients with visceral leishmaniasis; Andrade TM et al.; Bacterial infections are often seen in patients with visceral leishmaniasis . To determine the incidence of such infection and the more common infectious agents, 30 consecutive patients with visceral leishmaniasis were followed throughout hospitalization . There were 24 episodes of bacterial infection in 18 patients (60%) . The incidence of bacterial infections in these patients was 22.2/1000 days of admission . The proportion of patients becoming infected by time was significantly greater in the visceral leishmaniasis group than in controls (P less than .01) . The skin, respiratory tract, and middle ear were the most common sites of infection, and Pseudomonas aeruginosa and Staphylococcus aureus were the most common agents . Low-grade-virulence bacteria (e.g., Serratia and Providencia species) were also isolated from some cases . Bacterial infections (mainly nosocomial) in patients with visceral leishmaniasis tend to be severe and can cause death . When bacterial infection is suspected in these patients, empiric antibiotic therapy should be started immediately, including coverage for P . aeruginosa and S . aureus, after appropriate diagnostic procedures are taken. Eur J Clin Microbiol Infect Dis, 1990 Jun, 9(6), 435 - 8 In vitro activity of Ro 24-6392, a novel ester-linked co-drug combining ciprofloxacin and desacetylcefotaxime; Jones RN; In preliminary in vitro susceptibility tests Ro 24-6392 (desacetylcefotaxime with a 3'-ciprofloxacin substitution) demonstrated activity against a wide spectrum of aerobic bacteria, 98.6% of strains being inhibited by less than or equal to 8 mg/l . Of the commonly isolated species tested, only enterococcal strains were resistant (MICs greater than or equal to 32 mg/l) . The potency of Ro 24-6392 was generally between that observed for the two hydrolysis components, except in the case of Providencia stuartii and penicillin-resistant pneumococci isolates which were more susceptible to Ro 24-6392 . This new dual-action compound appears to be slightly more active than the previously studied peer drug Ro 23-9424, principally because of the superior activity of the fluoroquinolone component, ciprofloxacin. J Clin Microbiol, 1989 Sep, 27(9), 1969 - 72 Usefulness of trehalose fermentation and L-glutamic acid decarboxylation for identification of biochemically aberrant Providencia stuartii strains; Fischer R et al.; A total of 849 Providencia isolates were collected during a 4-year period when an increased incidence of nosocomial Providencia stuartii infection was noted in urologic wards . Of these isolates, 630 were identified as P . stuartii, 206 were identified as Providencia rettgeri, and 1 was identified as Providencia alcalifaciens . Twelve inositol-positive isolates from 10 patients (10 strains) resembled P . stuartii in fermenting trehalose but resembled P . rettgeri in fermenting D-arabitol or meso-erythritol or both . The latter traits, however, were not stable in all cases . These aberrant strains were identified as P . stuartii on the basis of their O antigens and DNA hybridization experiments . All isolates were tested for L-glutamic acid decarboxylase activity by a qualitative thin-layer chromatography method . All P . stuartii isolates, including the aberrant ones, were trehalose positive and L-glutamic acid decarboxylase negative . None of the P . rettgeri isolates fermented trehalose, while 99.0% of them and the single P . alcalifaciens strain were L-glutamic acid decarboxylase positive . Thus, trehalose fermentation and L-glutamic acid decarboxylation are more useful for separating P . stuartii from P . rettgeri than are D-arabitol and meso-erythritol fermentation. Microbiologica, 1989 Jul, 12(3), 247 - 55 Modification of norfloxacin inhibition of DNA gyrase induced by a 28 KDal DNA binding protein; Landini P et al.; We have previously studied a clinical isolate of Providencia stuartii which showed high levels of resistance to 4-quinolones, aminoglycoside and beta-lactam antibiotics (Landini et al., 1987) . DNA gyrase from this isolate was inhibited for 50% of activity at a concentration of 15 microM of norfloxacin, which is about 5-fold higher compared to the 50% inhibitory concentration for a standard DNA gyrase . It has been described that 4-quinolone inhibition of DNA gyrase is caused by their binding to DNA and by the distortion induced in DNA tertiary structure, and that affinity binding of 4-quinolones is different for DNAs in different structures . In order to detect whether the interaction between pAT 153 and a protein able to modify DNA tertiary structure could affect norfloxacin inhibitory concentrations for DNA gyrase we purified from the clinical isolate of Providencia stuartii a DNA binding protein of about 28 KDal which induces changes in supercoiling degree of DNA . Assays of DNA gyrase activity were performed on the complex pAT 153-DNA binding protein-norfloxacin . Results showed an increase from 15 microM to 20 microM of 50% norfloxacin inhibitory concentration for DNA gyrase when pAT 153 was complexed with the 28 KDal protein. J Appl Bacteriol, 1989 Mar, 66(3), 253 - 8 Interaction of the bisbiguanides chlorhexidine and alexidine with phospholipid vesicles: evidence for separate modes of action; Chawner JA et al.; Strains of Providencia stuartii with demonstrated resistance towards chlorhexidine did not show such resistance towards either of the related biguanide antiseptics, alexidine or vantocil . Alexidine promoted a significantly faster alteration in the permeability of Escherichia coli cell membranes towards various metal cations than chlorhexidine . Differential thermal analysis of various mixed lipid vesicles and pure phospholipid vesicles showed alexidine to share with vantocil the property of producing lipid phase separation and domain formation . It is suggested that the nature of the end-group on the biguanides affects the ability to produce lipid domains in cell membranes and this this might, in turn, affect activity and resistance patterns observed. Ann Inst Pasteur Microbiol, 1988 Nov-Dec, 139(6), 689 - 702 An evaluation of allozyme amino acid substitutions for the study of molecular relationships in Providencia strains; Goullet P et al.; Through analysis of molecular relationships in terms of amino acid substitutions, intra- and interspecies differentiations in Providencia alcalifaciens, P . stuartii and P . rustigianii were evaluated among the electrophoretic variants of three enzymes, L-malate dehydrogenase, acid phosphatase and esterase-beta a, chosen for their distinct pattern of polymorphism . For each enzyme, molecular relatedness among variants defined by two-dimensional electrophoretic profiles was examined through protein titration curves . P . stuartii strains appeared identical to each other and P . rustigianii strains were closely related, whereas the division of P . alcalifaciens strains into previously described zymotypes A1 and A2 was refined in molecular terms . A gradient of molecular interrelatedness between the species was observed for the three enzyme loci: with L-malate dehydrogenase, the three species appeared very closely related; with acid phosphatase, P . stuartii and P . alcalifaciens were more closely related to each other than to P . rustigianii; with esterase-beta a, P . alcalifaciens and P . stuartii appeared partially related, whereas no such relatedness was observed between these two species and P . rustigianii. J Antimicrob Chemother, 1988 Aug, 22(2), 105 - 11 The effects of clavulanic acid and sulbactam on beta-lactamase biosynthesis; Farmer TH et al.; Clavulanic acid and sulbactam were tested as inducers of beta-lactamase in 21 strains of Gram-negative bacteria . beta-Lactamase synthesis was inducible in all these strains, as demonstrated by the increased enzyme activities obtained when cells were grown in the presence of 10 mg/l of cefoxitin, the increase varying from 11- to 734-fold However, at the same concentration, neither inhibitor induced significant amounts of beta-lactamase, except in one strain of Providencia rettgeri, where both were potent inducers . When steps were taken to overcome the inhibitory effects of clavulanic acid and sulbactam, they were still found to be ineffective inducers . We conclude that, at therapeutic concentrations, clavulanic acid and sulbactam are generally poor inducers of beta-lactamase . Amoxycillin and ampicillin induced more beta-lactamase than the inhibitors, but were much less effective than cefoxitin, except for the strain of Prov . rettgeri. J Bacteriol, 1988 May, 170(5), 2202 - 7 Purification, characterization, and genetic organization of recombinant Providencia stuartii urease expressed by Escherichia coli; Mulrooney SB et al.; Recombinant urease from Providencia stuartii has been expressed in and purified from Escherichia coli, and the genetic organization of the structural genes has been determined . Urease expression was induced by urea and repressed by nitrogen-rich components in the medium . The urease protein was purified 331-fold by DEAE-Sepharose, phenyl-Sepharose, Mono-Q, and phenyl-Superose chromatographies with a 7.3% yield . The enzyme possessed a Km for urea of 9.3 mM and hydrolyzed urea at a Vmax of 7,100 mumol/min per mg . P . stuartii urease is composed of three polypeptides (Mrs, 73,000, 10,0000, and 9,000) denoted by alpha, beta, and gamma . The native enzyme is best described as (alpha 1 beta 2 gamma 2)2, based on a native Mr of 230,000, obtained by gel filtration chromatography, and on the Coomassie blue staining intensities of the individual subunits . Atomic absorption analysis of the pure protein revealed 1.9 +/- 0.1 nickel ions per alpha 1 beta 2 gamma 2 unit . In vitro transcription-translation analysis of transposon insertion mutants of the recombinant urease demonstrated that the urease peptides are encoded on adjacent DNA sequences and transcribed as a polycistronic mRNA in the order gamma, beta, and then alpha . Three urease-defective insertion mutants were identified that did not affect synthesis of urease subunit polypeptides, indicating that some nickel processing, enzyme activation, or other function may also be necessary for producing an active urease. J Clin Microbiol, 1988 Feb, 26(2), 323 - 7 Commercial identification systems often fail to identify Providencia stuartii; Cornaglia G et al.; We tested 145 clinical isolates in an attempt to evaluate some of the most widely used commercial identification systems in Europe in terms of their ability to identify Providencia strains . Two manual miniaturized systems (API 20E and Enterotube II) and three mechanized-automated systems (Cobas-Bact, Sceptor System, and Titertek-Enterobac-Rapid Automated System) were evaluated . Providencia alcalifaciens and Providencia rettgeri strains were correctly identified by all systems in all cases, and in most cases identification was achieved without the aid of supplementary tube tests . By contrast, Providencia stuartii was identified without the aid of supplementary tube tests for only 42.5% (API 20E), 37.5% (Enterotube), 68.7% (Sceptor), and 71.2% (Cobas-Bact) of the isolates . The overall misidentification rates were 16.3, 11.3, 11.3, and 10%, respectively . The Titertek-Enterobac-Rapid Automated System failed to identify only 1 of 80 strains (1.3%) and required supplementary tests in 2 other cases (2.5%) . Since four of the multitest systems examined often failed to correctly identify P . stuartii, we conclude that supplementary conventional tube tests should always be used to distinguish this species from the other taxa of the Proteeae tribe. J Appl Bacteriol, 1988 Jan, 64(1), 27 - 35 Numerical analysis of electrophoretic protein patterns of Providencia alcalifaciens strains from human faeces and veterinary specimens; Holmes B et al.; Twenty-five strains of Providencia alcalifaciens from various countries have been characterized by one-dimensional SDS-PAGE of cellular proteins . They comprised 15 from human faeces, one from duck faeces, one from a guinea-pig eye and eight from unknown sources . Also included, for reference purposes, were the type strains of three other Providencia species . The protein patterns, which contained 45-50 discrete bands, were highly reproducible and were used as the basis for two numerical analyses . In the first, in which the principal protein bands (in the 33-40 kD range) were excluded, the 25 Prov . alcalifaciens strains formed, at the 83% S level, a single cluster whilst the three Providencia reference strains remained unclustered . In the second, which included all the protein bands, the 25 Prov . alcalifaciens strains formed 10 clusters at the 85% S level . We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides the basis for typing clinical strains of Prov . alcalifaciens . Reference strains of each of the 10 PAGE types identified are available from NCTC for inclusion in future studies. Chemioterapia, 1987 Oct, 6(5), 324 - 8 Preliminary studies on the effect of glutathione S-transferase from Providencia stuartii on the antimicrobial activity of different antibiotics; Piccolomini R et al.; Glutathione S-transferase has been isolated and purified from a Providencia stuartii CH 114 strain . The effect of the enzyme on the antimicrobial activity of amikacin, cefotaxime, cephalexin, ampicillin, nalidixic acid, and ofloxacin was tested . The efficiency of all antibiotics tested, except nalidixic acid and ofloxacin, is relevantly decreased by the presence of glutathione S-transferase in the medium culture, as proved by the increased value of MIC . The effect of glutathione S-transferase on the drugs is not significantly affected by the addition of exogenous glutathione . The possible mechanism of action of the enzyme is also discussed. Microbiologica, 1987 Oct, 10(4), 403 - 11 Modifications of DNA-gyrase and of permeability in a norfloxacin-resistant clinical isolate of Providencia stuartii; Landini P et al.; We obtained a clinical isolate of Providencia stuartii showing a high level of resistance to norfloxacin and to other 4-quinolones, whose target is the enzyme DNA-gyrase . This strain showed resistance also to beta-lactam and aminoglycoside antibiotics . In order to detect modification of DNA-gyrase, we performed supercoiling assays in vitro in presence of norfloxacin and ciprofloxacin . Furthermore, outer membrane proteins, which are involved in permeability mechanisms, were analyzed on SDS-polyacrylamide gels . Results showed that both modifications in DNA-gyrase and changes in outer membrane proteins can be held responsible for resistance to 4-quinolones; moreover, these modification are probably supported by a third mechanism of resistance. J Gen Microbiol, 1987 Jul, 133 ( Pt 7), 1767 - 73 Fractionation of the Providencia stuartii cell envelope; Chopra I et al.; Cell envelopes (i.e . unfractionated inner and outer membranes) were obtained from Providencia stuartii by following procedures previously applied to the isolation of envelopes from Escherichia coli . The P . stuartii envelopes contained known inner membrane enzymes that included a variety of dehydrogenases and ATPase . The catalytic activity of the ATPase depended upon the concentration of magnesium ions, the substrate (ATP) level and the ratio of magnesium ions to ATP . Cell envelopes from P . stuartii were further fractionated to recover inner and outer membrane polypeptides by treatment with the detergent Sarkosyl . Proteins from the periplasmic region were recovered by a simple osmotic shock procedure also previously applied to E . coli . The purity of the various P . stuartii cell envelope fractions was assessed by a combination of techniques that included one- and two-dimensional gel electrophoresis of proteins, enzyme assays and detection of penicillin-binding proteins. South Med J, 1987 Jul, 80(7), 909 - 11 Nosocomial infectious balanoposthitis in neutropenic patients; Manian FA et al.; We have reported balanoposthitis as a source of fever and bacteremia in two neutropenic uncircumcised patients . The etiologic organisms were Pseudomonas aeruginosa in one case and Providencia stuartii in the other . In one patient, the diagnosis was delayed by the presence of a condom catheter . This emphasizes the importance of personal hygiene in uncircumcised men about to undergo immunosuppressive therapy, and the need for judicious use of condom catheters in such patients. J Antimicrob Chemother, 1987 Jun, 19(6), 743 - 51 Inhibition of Providencia stuartii cell envelope enzymes by chlorhexidine; Chopra I et al.; The possibility that chlorhexidine is a specific inhibitor of membrane bound bacterial adenosine triphosphatase (ATPase) was addressed . The in-vitro susceptibilities of several Providencia stuartii cell envelope enzymes, including ATPase, to chlorhexidine were compared . The following concentrations of chlorhexidine were required to cause 50% inhibition of enzyme activity in preparations from chlorhexidine-sensitive strains (MIC 50 mg chlorhexidine/l): ATPase (160 mg/l), succinic dehydrogenase (greater than 300 mg/l), penicillin binding protein 7 (300 mg/l) and beta-lactamase (45 mg/l) . Fifty per cent inhibition of the ATPase from a chlorhexidine-resistant strain (MIC 1600 mg/l) was achieved at an in-vitro concentration of 225 mg chlorhexidine/l . Our observations do not support the suggestion that bacterial membrane-bound ATPases are specific targets for chlorhexidine. Ukr Biokhim Zh, 1987 Jan-Feb, 59(1), 19 - 24 {Isolation and properties of aminocaprolactam hydrolase from Providencia alcalifaciens}; Galaev IuV et al.; L-alpha-aminocaprolactam hydrolase possessing the L-lysinamidase activity was isolated and purified from Providencia alcalifaciens . The purification procedure of enzymes included cell destruction on USDL-1, fractionation by ammonium sulfate, gel-chromatography on G-100, ion exchange chromatography on DEAE-cellulose . The purification resulted in a homogeneous enzyme which possessed the both activities . The enzyme molecular weight (180 kDa) was estimated by gel chromatography on Sephadex G-200 . Km was 3.5 mM in the phosphate buffer (pH 7.2) . L-alpha-aminocaprolactam hydrolase and L-lysinamidase may be related to metal-dependent enzymes requiring Mg++. J Antimicrob Chemother, 1987 Jan, 19(1), 27 - 30 Inducible expression of an aminoglycoside-acetylating enzyme in Providencia stuartii; Swiatlo E et al.; Strains of Providencia stuartii were collected from a chronic-care geriatric ward of a large Veteran's Administration hospital . Two strains of P . stuartii, and one of Escherichia coli isolated from the same ward, were transferred five times in the presence of gentamicin or netilmicin . At the fifth transfer the MICs of both gentamicin and netilmicin for the two P . stuartii strains had increased at least ten-fold . The MICs for the E . coli remained essentially unchanged . Enzyme assay demonstrated increased activity of an aminoglycoside-2'-acetylating enzyme in P . stuartii during the period of the transfers . Five subsequent transfers in the absence of aminoglycosides resulted in return of enzyme activity to the initial level . Neither of the P . stuartii strains tested contained any detectable plasmid DNA . The 2'-acetylating enzyme in some strains of P . stuartii can be induced to high levels of activity by exposure to gentamicin or netilmicin. J Appl Bacteriol, 1986 Nov, 61(5), 373 - 81 Reversal of the surface effects of chlorhexidine diacetate on cells of Providencia stuartii; Ismaeel N et al.; Chlorhexidine diacetate (CHA) increased the hydrophobicity of the cell surface of cells of three strains of Providencia stuartii . Removal of at least some of the CHA from the cells by washing them with an appropriate antidote partially reversed the hydrophobicity-increasing action of the biguanide . The effects of other treatments on cell surface hydrophobicity were examined with these strains and, for comparison, with two strains each of Escherichia coli and Pseudomonas aeruginosa: ethylenediamine tetraacetic acid affected all strains, although not to the same extent, whereas thermal injury (55 degrees C) produced marked changes only with the two E . coli strains. Arch Microbiol, 1986 Apr, 144(3), 213 - 8 Lipopolysaccharide of Providencia rettgeri . Chemical studies and taxonomical implications; Basu S et al.; The chemical constitutional analysis of the lipopolysaccharide (LPS) isolated from Providencia rettgeri was carried out . Polyacrylamide gel electrophoresis using sodium dodecylsulfate or sodium deoxycholate showed that the lipopolysaccharide mostly consisted of short sugar chains . The lipid A was precipitated out after mild acid hydrolysis of LPS . From the supernatant degraded polysaccharide and unsubstituted core fractions were isolated . Compositional analysis of the core material revealed the presence of galacturonic acid, galactose, glucose, glucosamine, L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid, alanine and phosphorus . Methylation analysis of the core material indicated the presence of terminal units of glucose, galacturonic acid and glucosamine . The chemical structure of the lipid A was elucidated . It constitutes a beta-1,6-glucosamine disaccharide substituted on either side by ester and glycosidically-bond phosphate residues . The ester-bound phosphate was found to be substituted by a 4-amino-4-deoxy-L-arabinosyl residue . The amino groups of the backbone disaccharide are N-acylated by 3-O-(14:0)14:0 and 3-O-14:0 . Two hydroxyl groups of the disaccharide are esterified by 3-O-(14:0)14:0 and 3-O-14:0 . The taxonomical importance of these structural details will be discussed. J Appl Bacteriol, 1986 Apr, 60(4), 361 - 7 Resistance of Providencia stuartii to chlorhexidine: a consideration of the role of the inner membrane; Ismaeel N et al.; Spheroplasts of three strains of Providencia stuartii (one sensitive, one moderately sensitive and one resistant to chlorhexidine) were induced by cefoxitin, glycine or a lysozyme-tris-EDTA combination, and their susceptibility to chlorhexidine-induced lysis investigated . Maximum lysis of spheroplasts occurred at a low (2.5-5 micrograms/ml) concentration of chlorhexidine and was greatest with the most sensitive strain, Pv2 . The possible role of the inner membrane in chlorhexidine resistance is considered in the light of the findings obtained. Arch Surg, 1986 Jan, 121(1), 31 - 6 Effects of environment on infection in burn patients; Shirani KZ et al.; Burn patients in an early cohort (n = 173) treated in an intensive care ward without separate enclosures were compared with a later cohort (n = 213) treated in a renovated unit with separate bed enclosures . The number of patients developing infection was significantly reduced in the late group . Observed mortality was compared with mortality predicted on the basis of burn size and age alone . Reduction in observed compared with predicted mortality, inapparent in the early group, was seen in the late group and was restricted to the subgroup of patients with predicted mortality of 25% to 75%, in which the observed mortality of 28.3% was less than the predicted mortality of 48.7% . The incidence of infected patients was reduced from 58.1% in the early cohort to 30.4% in the late cohort . In comparison of the early cohort with the late cohort, the overall proportion of patients with bacteremia was reduced from 20.1% to 9.4%, while the incidences of both pneumonia and burn wound invasion remained unchanged . Providencia and Pseudomonas species, endemic in the early cohort, were eliminated in the late cohort . Reduction of infection by environmental manipulation in burn patients was possible and was associated with improved survival. J Antimicrob Chemother, 1985 Dec, 16(6), 685 - 9 Antiseptic-induced changes in the cell surface of a chlorhexidine-sensitive and a chlorhexidine-resistant strain of Providencia stuartii; el Moug T et al.; The effects of chlorhexidine diacetate and benzalkonium chloride on the cell surface of a chlorhexidine-sensitive (Pv 2) and a chlorhexidine-resistant (Pv 67) strain of Providencia stuartii are described . Low concentrations of chlorhexidine diacetate (10 mg/l and upwards) increased the hydrophobicity of Pv 2, whilst having little effect on Pv 67 . Both strains were resistant to benzalkonium chloride but a concentration as low as 2 mg/l induced a significant increase in hydrophobicity in Pv 2, with 25-50 mg/l needed to induce a similar type of increase in Pv 67 . The possible nature of the resistance is discussed. Ann Inst Pasteur Microbiol, 1985 May-Jun, 136A(3), 347 - 58 {Electrophoretic polymorphism of lactate, malate and glutamate dehydrogenases, acid phosphatase and esterases of Providencia alcalifaciens, P . stuartii and P . rustigianii}; Goullet P et al.; The polymorphism of glutamate, lactate and malate dehydrogenases, of acid phosphatase and of esterases of 27 strains of Providencia alcalifaciens, 35 strains of P . stuartii and 17 strains of P . rustigianii was investigated by conventional electrophoresis in polyacrylamide agarose gel and by isoelectric focusing in thin-layer polyacrylamide gel . For each enzyme analysed, the three species were characterized by a distinct electrophoretic pattern . The number of allozymes detected by conventional electrophoresis was greater than that detected by isoelectric focusing . The use of these two techniques in parallel led to improved detection of polymorphism of esterase alpha beta from P . alcalifaciens . A two-dimensional profile obtained by plotting isoelectric points against electrophoretic mobilities for malate dehydrogenase, acid phosphatase and beta-A esterase showed a molecular relationship between the diverse allozymes and demonstrated their taxonomic values . Polymorphism varied considerably according to the enzyme and species analysed and was correlated with DNA heterogeneity . The strains of P . alcalifaciens exhibited the greatest enzyme polymorphism and were classified into two main zymotypes reflecting genetic divergence within this species, whereas the strains of P . stuartii were electrophoretically less variable. J Gen Microbiol, 1985 Apr, 131 ( Pt 4), 927 - 33 Evolution of an R plasmid from a cryptic plasmid by transposition of two copies of Tn1 in Providencia stuartii; Hawkey PM et al.; Examination of a series of isolates of Providencia stuartii collected over an 18 month period from a chronic-care patient at Bristol Royal Infirmary revealed the emergence of resistance to carbenicillin . Resistance was mediated by a 47 kb plasmid which transferred by conjugation to a plasmid-free strain of P . stuartii but not to Escherichia coli . Carbenicillin-sensitive isolates were either plasmid-free or contained a 36 kb cryptic plasmid . Restriction endonuclease mapping of this plasmid showed it to be closely related to 32 kb and 34 kb cryptic plasmids reported previously in P . stuartii from Bristol . Mapping of the R plasmid showed it to be derived from the 34 kb cryptic plasmid by transposition of two copies of Tn1. Mol Gen Genet, 1985, 200(3), 476 - 81 Comparative studies on the structural gene for the ribosomal protein S1 in ten bacterial species; Schnier J et al.; By applying the Southern blot technique we compared the structural gene rpsA for ribosomal protein S1 and its preceding sequence from Escherichia coli with nine other bacterial species . We found high homology among the structural genes of E . coli and other gram-negative but not gram-positive bacteria . In contrast, the regulatory sequence preceding the structural gene was not highly conserved among the organisms studied . Cloning and DNA sequence analysis of a 1.2 kb fragment coding for most of the structural gene for S1 from Providencia localized some strongly conserved parts of the DNA sequence, despite the fact that the codon usage showed considerable divergence from that of E . coli. J Med Microbiol, 1984 Oct, 18(2), 277 - 84 Cryptic plasmids in hospital isolates of Providencia stuarti; Hawkey PM et al.; The distribution of cryptic plasmids among 123 isolates of Providencia stuarti from a hospital ward during a prospective epidemiological study is reported . Two closely related stable plasmids (34 Kb and 36 Kb) were identified by restriction endonuclease digest analysis of plasmid DNA . One or other of these cryptic plasmids was carried by 40 isolates, the remainder were plasmid-free . A higher proportion of one cryptic plasmid (CPT-A) was found in environmental isolates than in isolates from patients . The serotype of all isolates of P . stuarti was O63 and they were epidemiologically related . Two of the eight patients colonised by P . stuarti carried all three possible variants: plasmid-free (PFI) strains or strains containing cryptic plasmid A (CPT-A) or cryptic plasmid B (CPT-B) . The epidemiological significance of these results is discussed. Antimicrob Agents Chemother, 1984 Aug, 26(2), 187 - 91 In vitro and in vivo antibacterial activities of K-4619, a new semisynthetic aminoglycoside; Saino Y et al.; The antibacterial activities of K-4619 (3-de-O-methylsporaricin A sulfate) were compared with those of sporaricin A, amikacin, dibekacin, and gentamicin . K-4619 exhibited a high order of activity against gram-positive and gram-negative bacteria, including Pseudomonas aeruginosa . Its activity against Providencia species and Serratia marcescens was the highest of all drugs tested . K-4619 was highly effective against bacteria that produce various aminoglycoside-inactivating enzymes, except for 3-acetyltransferase I . The bactericidal activity of K-4619 was somewhat greater than that of amikacin . The activity of K-4619 against gram-negative bacteria increased at alkaline pH and was hardly affected by inoculum size, addition of horse serum, and composition of the medium . The in vivo protective effect of K-4619 against infections with Klebsiella pneumoniae, S . marcescens, and P . aeruginosa in mice was greater than that of sporaricin A . K-4619 was also active in mice infected with gentamicin- or amikacin-resistant strains bearing some of the aminoglycoside-inactivating enzymes. Zh Mikrobiol Epidemiol Immunobiol, 1984 Jun, (6), 44 - 7 {Serological passage of bacteria of the genus Providencia isolated from patients with wound, urological and intestinal infections}; Raginskaia VP et al.; A total of 395 Providencia strains isolated from patients and the environment were studied . Their differentiation by O and H antigens revealed that these strains belonged to 36 serological groups and contained 26 varieties of the flagellar antigen . In accordance with the combinations of O and H antigens, 59 serological variants were found among the Providencia strains . 33 new, formerly unknown serovars and 2 flagellar antigens were added to the existing antigenic and diagnostic scheme of Providencia . Most of the strains isolated from patients with surgical complications and burns were found to belong to P . stuartii, serovars O44: H4 and O4: H4, and from patients with acute intestinal diseases, to P . alcalifaciens, serovars O2: H26 and O50: H33. Antimicrob Agents Chemother, 1983 Sep, 24(3), 347 - 52 Resistance of gram-negative bacilli as related to hospital use of antimicrobial agents; Ma MY et al.; The development of resistance of gram-negative bacilli, which are common nosocomial pathogens, is an increasing problem . It is generally accepted that this resistance may directly reflect the frequency of use of various antimicrobial agents . Because our institution experienced in 1976 a dramatic change in the pattern of antimicrobial use, primarily a marked decrease in prescribing cephalosporins, we attempted to evaluate retrospectively the effects of this change upon the resistance of gram-negative bacilli that are common nosocomial pathogens . Susceptibilities of Klebsiella and Providencia spp., Pseudomonas aeruginosa, and Serratia marcescens were determined for the years 1975 to 1979 . Not unexpectedly, we observed a substantial decrease in cephalosporin resistance . An unexpected finding was a decrease in aminoglycoside resistance, despite increased use of these agents . The possibility that decreased cephalosporin use may lead to decreased aminoglycoside resistance is an intriguing and provocative thesis which can only be speculative at this time but which would seem worthy of additional formal investigation. J Clin Microbiol, 1983 Aug, 18(2), 227 - 35 Epidemiology of gentamicin-resistant, gram-negative bacillary colonization in a spinal cord injury unit; Shlaes DM et al.; A prospective epidemiological survey of a spinal cord injury unit for gentamicin-resistant, gram-negative bacilli was undertaken . The initial survey of the unit suggested a low level of cross-infection involving Pseudomonas aeruginosa and Providencia stuartii . However, a longitudinal study of new admissions revealed that only 13 of 52 nosocomial acquisitions could be considered to be due to cross colonization . Comparison of data on antibiotic use did not suggest selective pressure for resistant endogenous flora . Nosocomial acquisition was directly related to the length of the hospital stay . Antibiotic susceptibility testing of gentamicin-resistant, gram-negative bacilli showed only minor differences between nosocomial isolates and those present during the initial survey . Of the usual antimicrobial agents, amikacin, carbenicillin, and cefoxitin were the most active against all organisms, with the exception of Serratia spp . Of the new beta-lactams, ceftazidime and imipemide (N-formimidoyl thienamycin) were most active. J Hyg (Lond), 1983 Jun, 90(3), 475 - 88 Plasmid pattern analysis of natural bacterial isolates and its epidemiological implication; Tietze E et al.; Natural isolates of Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae, and Providencia stuartii were analysed to determine their plasmid content . This data allowed the identification of nosocomial strains of K . pneumoniae and P . stuartii and helped in the differentiation of epidemic strains of E . coli 0111 and S . typhimurium . Phenotypically similar isolates of S . typhimurium could be shown to be of independent origin using plasmid pattern analysis . The dissemination of a particular plasmid through different strains of S . typhimurium resulted in a simulation of a very widely distributed epidemic strain, because the plasmid interfered with the phage type of its host strain in addition to determining resistance properties . Plasmid pattern analysis disclosed two independently existing but interacting epidemic processes: a bacterial 'epidemic' strain may become disseminated over a large territory and may predominate there for a long time; a single plasmid, however, may also become distributed through many different bacterial strains and may spread over a large territory . Plasmid pattern analysis provides a valuable and universal epidemiological laboratory method. Antimicrob Agents Chemother, 1983 Apr, 23(4), 619 - 21 Comparative in vitro activity of semisynthetic penicillins against Proteeae; Hawkey PM et al.; The in vitro susceptibilities of 181 isolates of precisely identified Proteeae species to five semisynthetic penicillins were determined with low and high inocula . Significant differences in susceptibility patterns among various Proteeae species to the penicillins examined were demonstrated . Providencia stuartii was clearly distinguished from Providencia alcalifaciens by its greater resistance to the antibiotics tested. Antibiotiki, 1982 Feb, 27(2), 130 - 3 {Effectiveness of the combined use of tobramycin with carbenicillin or cephalosporins in experimental infection in mice caused by Providencia stuartii}; Shapovalova SP et al.; Antibacterial activity of tobramycin in combination with carbenicillin or cephalosporins against 20 strains of Providencia stuartii was studied . The combinations of tobramycin with carbenicillin (1 : 2.5) or cephaloridine (1 : 1) were most active . The MIC of tobramycin used in combination with carbenicillin or cephaloridine decreased 3-30 times . In treatment of albino mice with sepsis caused by Providencia stuartii it was possible to lower 2-8 times the dose of tobramycin used in combination with carbenicillin or cefazolin. Mikrobiologiia, 1982 Jan-Feb, 51(1), 54 - 9 {Characteristics of the fatty acid composition of Providencia alcalifaciens and Providencia stuartii lipids}; Vasiurenko ZP et al.; The fatty acid composition of total cellular lipids and lipid A of lipopolysaccharides was studied in Providencia alcalifaciens and P . stuartii . Significant differences were found in the fatty acid composition of total cellular lipids from the above species; these are caused mainly by the presence of cyclopropane fatty acids (methylenehexadecanoic and methyleneoctadecanoic acids) in P . stuartii and their absence in P . alcalifaciens . Differences were also established in the content of fatty acids in lipid A of lipopolysaccharides from P . alcalifaciens and P . stuartii . The data are consistent with the idea that P . alcalifaciens and P . stuartii are separate taxons though, apparently, closely related. Antimicrob Agents Chemother, 1981 Jul, 20(1), 115 - 9 Examination of major disagreements in susceptibility test results by Autobac-1 and MS-2; Lindsey NJ et al.; Major disagreements between two automated susceptibility test instruments, Autobac-1 and MS-2, generated from 4,213 tests on 630 bacterial isolates, were analyzed . The major disagreement rate between the instruments was only 7.4% . The highest major disagreement rates for bacteria were with Staphylococcus epidermidis, enterococci, gram-positive bacilli, and Providencia species, and those for antibiotics were with ampicillin and penicillin G . With most other bacteria and antibiotics, the instruments disagreed at a rate less than 10% and frequently at a rate less than 4% . However, 32 specific bacterium-antibiotic combinations exceeded a 10% rate . Possible reasons for some of these higher disagreement rates are discussed . Reconciliation of instrument results with the standard agar disk diffusion result for each major disagreement revealed that about 15% were irreconcilable and, of the remaining 259, 177 agreed with MS-2 and 82 agreed with Autobac-1 . Major disagreements between instruments seemed random, but there appeared to be tendencies, with certain bacteria and certain antibiotics, for MS-2 to detect susceptible reactions more consistently than resistant ones and for Autobac-1 to detect resistant reactions more consistently than susceptible ones. Immun Infekt, 1981 Jul, 9(4), 150 - 3 {Investigation on the antimicrobial activity of cefoperazone (author's transl)}; Primavesi CA; Cefoperazone has the usual antimicrobial spectrum of the cephalosporins, but the ratios of resistance to the various bacterial strains - even if they are problematic-are very low . It remains to be seen, however, whether a widespread use of the drug induces an increase in the number of resistant strains . An outstanding advantage of cefoperazone is its efficacy against Pseudomonas aeruginosa; only one of 75 strains investigated was resistant . In the case of Providencia, an important germ in the field of urology that is often difficult to be affected by antibiotics, we could not find any resistant strains . We did, however, find moderate sensitive strains . Concerning Enterococci we did not find any resistant strains, but 96% of the strains were classified as moderate sensitive . That leads to the conclusion that the drug should only be effective in high dosage, when Enterococci are found . Even multi-resistant bacterial strains still respond considerably to cefoperazone . This applies in particular to Pseudomonas aeruginosa. J Clin Microbiol, 1981 Jun, 13(6), 1099 - 104 Antibiotic resistance in Providencia stuartii isolated in hospitals; McHale PJ et al.; A total of 238 isolates of Providencia stuartii obtained from infected patients in six Dublin hospitals were grouped by using serological and bacteriocin typing methods and tested for sensitivity to a number of antimicrobial agents . Most isolates were resistant to several of these agents . Resistance to tetracycline, resistance to penicillin, resistance to polymyxin, and probably resistance to nitrofurantoin was intrinsic . Plasmid screening coupled with resistance transfer studies showed that both chromosome-encoded and plasmid-coded resistance mechanisms were clinically important . Ampicillin resistance was both chromosomally and plasmid encoded, whereas resistance to kanamycin and resistance to carbenicillin were exclusively plasmid encoded . Gentamicin resistance was more common than kanamycin resistance, and although gentamicin-resistant strains contained aminoglycoside acetyltransferase activity, no association could be demonstrated with plasmid deoxyribonucleic acid in the strains tested . Unlike minimal inhibitory concentrations for kanamycin, minimal inhibitory concentrations for gentamicin varied over a wide range . P . stuartii isolated obtained from several different countries were tested for comparison . As a group, these strains were less resistant, but they did exhibit similar resistance properties. Vet Pathol, 1981 Apr, 18(Suppl 6), 59 - 69 Renal pathology of catheterized baboons (Papio cynocephalus); Heidel JR et al.; Renal disease was found at necropsy in 27 of 60 catheterized baboons (Papio cynocephalus) . Primary diagnoses were infarcts in eight baboons, septic embolic nephritis in four, and mesangioproliferative glomerulonephritis in 15 . Infarcts and septic embolic nephritis are multifocal renal diseases; mesangioproliferative glomerulonephritis is diffuse, progressive and often accompanied by severe proteinuria, hypoalbuminemia (less than 3 g/dl), and generalized edema . Five of the 15 baboons with glomerulonephritis also developed uremia and died of renal failure . Ten baboons with glomerulonephritis were given microbiological examination during their clinical course . Bacteria were isolated from the peripheral blood of eight baboons and from the catheter itself in two . The organisms isolated (in decreasing order of frequency) were Herellea sp., Streptococcus sp., Klebsiella sp., Staphylococcus sp., and Providencia sp . Prospective immunofluorescence studies were done on frozen sections from six catheterized baboons with glomerulonephritis and two uncatheterized controls . There were granular deposits of IgG in all six, IgM in five, C3 in four, and IgA and C4 in two . In the controls, only trace amounts of IgM were seen in one animal . The amount of IgG deposited in th glomeruli correlated with the severity of the glomerulonephritis . Bacterial antigens were found by immunofluorescence in glomeruli of three of five baboons examined . Glomerulonephritis in these baboons seems to be immunologically mediated, and in origin, pathogenesis and morphologic expression resembles shunt nephritis in humans with infected indwelling shunts and catheters. J Bacteriol, 1981 Apr, 146(1), 404 - 8 Hemagglutinins and fimbriae of Providencia spp; Old DC et al.; A series of 25 strains of Providencia spp . produced at least five different patterns of hemagglutination as judged by results from hemagglutination tests in the presence and absence of D-mannose and at least six distinct types of fimbriae as seen from observations with the electron microscope. J Clin Microbiol, 1981 Mar, 13(3), 561 - 5 Transferable urease activity in Providencia stuartii; Grant RB et al.; Six urea-positive Providencia stuartii strains were tested for transmissible urease determinants . Two strains, when implanted with "helper" conjugative plasmids, were found to be capable of transferring urease genes to Escherichia coli or urea-negative P . stuartii . Recombination of the urease genes with the helper plasmid in P . stuartii was noted in one case . One of the urea-positive P . staurtii strains was found to harbor a conjugative plasmid which mediated both urease activity and ampicillin resistance . This large plasmid (molecular weight, approximately 140 x 10(6)) was transmissible to and stably maintained in E . coli strains . The demonstration of transmissible genes for urease activity in P . stuartii is significant in that it accounts for previous problems associated with classifying urea-positive strains of this species. Proc Natl Acad Sci U S A, 1981 Mar, 78(3), 1503 - 7 Cloning and expression of the Pst I restriction-modification system in Escherichia coli; Walder RY et al.; Here we report the cloning and preliminary characterization of the Pst I restriction-modification system of Providencia stuartii 164 . Transformants of Escherichia coli carrying the Pst I gene system inserted into the cloning vector pBR322 were selected on the basis of acquired resistance to bacteriophage lambda infection . Pst I endonuclease was detected in osmotic shock fluid from each of the resistant clones . Plasmid and chromosomal DNA from these clones could not be digested by Pst I, indicating that the gene for the corresponding modification enzyme had also been cloned and was being expressed . The smallest recombinant plasmid encoding both activities, pPst201, contains an insert of approximately 4000 base pairs . In vitro transcription studies indicate that this DNA fragment also contains the endogenous promoter(s) of the system . When pPst201 was introduced into a minicell-producing strain of E . coli, two new proteins, 32,000 and 35,000 daltons, were synthesized . We have assigned these to the Pst I modification (methylase) and restriction enzymes, respectively . The active form of the restriction enzyme is a dimer, as determined by gel filtration . Constructed transformants of P . stuartii 164 that carry the Pst I system inserted into pBR322 produce approximately 10 times more Pst I endonuclease activity than does the native strain. Antimicrob Agents Chemother, 1980 Dec, 18(6), 868 - 71 Differences among Providencia species in their in vitro susceptibilities to five antibiotics; Penner JL et al.; Significant differences among the three Providencia species (P . alcalifaciens, P . stuartii, and P . rettgeri) in antimicrobial susceptibilities to five antibiotics were shown . P . stuartii was the most resistant of the three species, and P . alcalifaciens was the most susceptible . P . rettgeri was similar to P . stuartii in susceptibilities to cefoxitin, cephalothin, and cefamandole but differed in showing greater susceptibilities to tobramycin and gentamicin . Cefoxitin (16 micrograms/ml) and cefamandole (16 micrograms/ml) inhibited a greater proportion of P . stuartii isolates than did cephalothin, tobramycin, or gentamicin . The susceptibilities of urea-positive isolates of P . stuartii resembled more closely the susceptibilities of urea-negative isolates of this species than those of P . rettgeri isolates, a finding consistent with the recent recommendation for transferring such urea-positive strains to P . stuartii . Among P . stuartii isolates, marked resistance to cefoxitin accompanied by susceptibility to cefamandole was predominantly restricted to isolates of one serotype (O55) . The use of isolates that had been serotyped and classified according to recent proposals for taxonomic changes in the Proteeae provided for clearer demonstration of species differences in susceptibility. Nucleic Acids Res, 1980 Nov 11, 8(21), 4943 - 54 A comparison of DNA cleavage by the restriction enzymes SalPI and PstI; Carter JA et al.; Methods for obtaining highly active, exonuclease-free, stable preparations of the Streptomyces albus P restriction enzyme SalPI are described . SalPI and its isoschizomer PstI (from the taxonomically distant Providencia stuartii 164) both cleave their recognition sequence (5'-CTGCAG-3') to generate fragments terminating in tetranucleotide 3' extensions whose sequence is 5'-TGCA-3' . Bacteriophage R4G2 DNA, protected against SalPI cleavage by pregrowth on S . albus P, is also protected against PstI cleavage; and total DNA of both S . albus P and P . stuartii 164 is resistant to cleavage by both enzymes. Zh Mikrobiol Epidemiol Immunobiol, 1980 May, (5), 100 - 3 {Serotyping of bacteria of the genus Providencia using the indirect hemagglutination test and the indirect hemagglutination inhibition test}; Levina LA et al.; The experiments with the collection of strains and diagnostic sera available in the USSR confirmed the data contained in the literature on the good prospects of using the indirect hemagglutination test for the serotyping of bacteria belonging to the genus Providencia . Certain differences were observed in cross reactions, revealed by the indirect hemagglutination test and by agglutination: the interrelations of serogroups O14 and O25, O23 and O25, O23 and O29, O29 and O45, O37 and O49, determined by means of the indirect hemagglutination test, could not be detected in the same sera by the slide and tube agglutination tests . For the first time the usefulness of aqueous-saline extracts of agar cultures in the indirect hemagglutination inhibition test to specify more precisely the interrelations between individual bacterial serogroups of the genus Providencia has been shown. J Clin Microbiol, 1979 Dec, 10(6), 761 - 5 O-serotyping Providencia alcalifaciens; Penner JL et al.; The O-serotyping scheme for Providencia was tested on Providencia alcalifaciens isolates collected mostly from two hospitals . The specificites of the somatic (O) antigens of P . alcalifaciens were found to be different from those of Providencia stuartii, and separation of the Providencia typing scheme to allow separate typing of each species led to more efficient typing . All but 4 of 86 isolates were typable . Eighteen serotypes occurred among 53 typable isolates obtained from a pediatric hospital, and 11 occurred among 19 isolates from a general hospital . Thirty-two percent of the isolates from the pediatric hospital belonged to serotype O3, the most frequently isolated and most widely distributed type . The use of the serotyping scheme for P . alcalifaciens is advocated for further studies to examine strains of the species for enteropathogenic types. Infection, 1979, 7(3), 109 - 12 The sensitivity of gentamicin-resistant gram-negative bacilli to cefotaxime, other cephalosporins and aminoglycosides; Stephens M et al.; The sensitivities of 80 gentamicin-resistant gram-negative bacilli to cefotaxime, cefuroxime, cefoxitin, cefamandole, cefazolin, tobramycin, netilmicin and amikacin were determined . Amikacin was the most active amino-glycoside . However, the percentage sensitivity to cefotaxime of most of the species was higher than, or equal to any of the other antibiotics tested . Cefotaxime was particulary active against Providencia spp., Serratia spp., Klebsiella spp., and Pseudomonas maltophilia, being 16 to 256 times more active than the next best cephalosporin or cephamycin . Clinical trials of cefotaxime are now required. J Clin Microbiol, 1979 Jan, 9(1), 11 - 4 O serotyping of Providencia stuartii isolates collected from twelve hospitals; Penner JL et al.; A collection of 829 isolates of Providencia stuartii, mostly from urological specimens of patients in 12 hospitals, were O serotyped . Hospitals varied in serotype distribution, but most isolates (97%) fell into one or another of 14 O types of P . stuartii . One type (O63) was found in 10 hospitals, and six types (O4, O17, O25, O52, O55, O56) were found in 5 or more hospitals . These seven types were more common than others and included 753 (91%) of the isolates . Only four isolates agglutinated in Providencia alcalifaciens antisera and, for increased efficiency in serotyping, it is recommended that separate schemes be employed for P . stuartii and P . alcalifaciens . Strains endemic in different hospitals may differ in serotype and give rise to nosocomial infections that are clinically recognizable when infections occur in obvious clusters . Nosocomial infections occurring in low frequency among patients not located close to each other in the hospital may be detected with the aid of serotyping. Acta Microbiol Acad Sci Hung, 1979, 26(1), 55 - 62 {The anodically moving thermolabile antigen (ATA) of Gram negative bacteria . Protection of mice by immunization with ATA (author's transl)}; Seltmann G et al.; Mice are protected against lethal doses of Salmonella typhi-murium by vaccination with ATA extracted from Providencia . No protective effect is observed when the S . typhi-murium strains are enterotoxigenic . Thus protection is antibacterial, but not antitoxic. Can J Microbiol, 1978 Oct, 24(10), 1153 - 7 Simultaneous production of two types of beta-lactamase in Escherichia coli and Providencia stuartii; Letarte R et al.; The production of beta-lactamase has been studied in two strains isolated from clinical samples: Providencia stuartii MULB 501 and Escherichia coli MULB 130 . These strains were selected for their high resistance level to penicillins and cephalosporins . Determination and identification of beta-lactamase activity were achieved by combining several up-to-date methods including (i) neutralization by anti-beta-lactamase sera, (ii) purification by affinity chromatography on cephalosporin C linked Sepharose 4B, (iii) determination of substrate specificity and kinetic values (Km and Vmax) by a computerized microacidimetric method, and (iv) isoelectric focusing . The results clearly demonstrate that in these two strains there is a simultaneous production of different beta-lactamases: the first one is similar to the TEM penicillinase and the second one shares a typical cephalosporinase profile . This double beta-lactamase production is a relatively rare phenomenon. Mol Biol Rep, 1978 Jun 16, 4(2), 73 - 7 Location of the cleavage sites on the SV 40 DNA map produced by the restriction endonucleases Pst 1 and Bam 1; Fodor I; Restriction endonucleases from Providencia stuartii (Pst 1) and Bacillus amyloliquefaciens H (Bam 1) cleave SV 40 DNA at two and one specific sites, respectively . Using EcoRI and Hind III endonuclease restriction sites as reference, the two Pst I sites were mapped at 0.050; 0.265 and the Bam I site was mapped at 0.170 of the genome length, clockwise, from the single EcoRI cleavage site. Zentralbl Bakteriol {Orig A}, 1978 Apr, 240(2), 202 - 7 Bacteriocine typing of Providencia isolates; Al-Jumaili IJ et al.; A method of typing isolates of Providencia is described based on the sensitivities of the organisms to bacteriocines . Twelve standard bacteriocine producing strains were selected from a large number of isolates tested and from these liquid bacteriocine preparations were obtained . The activities of these preparations on over 300 isolates were determined and from this information a bacteriocine typing system has been developed. J Int Med Res, 1978, 6(5), 409 - 13 Susceptibility of 327 clinical isolates to netilmicin; Digranes A et al.; Netilmicin, a semisynthetic derivative of sisomicin, was tested against 327 isolates of Staphylococcus aureus, Pseudomonas aeruginosa and Gram-negative enteric bacilli . Seventy-two per cent of the isolates were inhibited at a concentration of 0.5 microgram netilmicin per ml, and 93% of the isolates were susceptible to 4 microgram per ml or less . The MICs of netilmicin and gentamicin for 24 Providencia and 38 Pseudomonas isolates were compared . The activity of netilmicin closely paralleled that of gentamicin, 46% of the Providencia isolates and 32% of the Pseudomonas isolates not being inhibited by 4 microgram per ml of either drug. Zh Mikrobiol Epidemiol Immunobiol, 1977 Dec, (12), 20 - 3 {Etiologic role of bacteria of the genus Providencia in acute intestinal diseases}; Kholodkova EV et al.; A study was made of morphological, cultural, biochemical properties and antigenic structure (by O-antigen) of 59 strains isolated in group acute intestinal affection of children at the pioneer camp . By the combination of biochemical properties (31 tests) all the cultures isolated were referred to Providencia alcalifaciens of biotype 7 . Serological typing with the aid of experimental diagnostic agglutinating O-sera showed these strains to be referred to serological group O2 . Identity of the O-antigen of the isolated cultures to the standard O2 strain was confirmed by the results of cross reactions of agglutinin absorption . The results of bacteriological and clinico-epidemiological studies offered a possibility of regarding the isolated Providencia alcalifaciens of serological group O2 as the causative agents of the group intestinal disease in children. Nucleic Acids Res, 1977 Jun, 4(6), 1989 - 98 A site-specific endodeoxyribonuclease from Streptomyces albus CMI 52766 sharing site-specificity with Providencia stuartii endonuclease PstI; Chater KF; A class II site-specific endodeoxyribonuclease (SalPI) was identified in cell-free extracts of Streptomyces albus CMI 52766 after high speed centrifugation and fractionation through Bio Gel AO.5M . SalPI cleaves lambda DNA into at least 18 fragments . Five cleavage sites were located in the linear lambda map by the use of double and triple restriction enzyme digests involving EcoRI, HindIII, SalGI and another new Streptomyces enzyme, SacI . The results were indistinguishable from those previously obtained for a Providencia stuartii enzyme, PstI, by Smith, Blattner & Davies (Nucleic Acids Res . 1976 3, 343) . SalPI and PstI were shown by a double digest test to have the same site specificity . None of 34 phages tested was obviously restricted by S . albus CMI 52766, and correspondingly DNA from two of them was not cleaved in vitro by SalPI . DNA from Streptomyces phage that does not form plaques on S . albus CMI 52766, and plasmid SCP2 DNA from S . coelicolor A3 (2), were both cleaved. J Infect Dis, 1976 Nov, 134 SUPPL, S297 - 307 Susceptibility of recently isolated bacteria to amikacin in vitro: comparisons with four other aminoglycoside antibiotics; Finland M et al.; In vitro tests for susceptibility to amikacin and to four other aminoglycoside antibiotics were carried out with strains of many bacterial species by use of an agar dilution method and an inocula replicator . In general, amikacin was as active as or more active against most of the organims than kanamycin, neomycin, and streptomycin; in particular, amikacin was active against strains resistant to one or more of these three antibiotics . Amikacin was more active than gentamicin against strains of Nocardia asteroides and Providencia stuartii and also against gentamicin-resistant strains of some other gram-negative bacilli, notably Serratia marcescens . However, gentamicin was more active than amikacin against most of the other gram-positive and gram-negative bacteria that were tested . In comparative tests of four media, minimal inhibitory concentrations MICs) were greater in tests with Mueller-Hinton agar, and generally somewhat lower in those with heart infusion agar, than in tests with trypticase soy agar and nutrient agar . Inocula of a 1:1,000 dilution of culture generally gave MICs lower than those obtained with undiluted cultures; the differences were small with enterococci, but they were greater with amikacin than with gentamicin in tests on strains of Klebsiella pneumoniae . These findings generally confirm those previously reported by others. Mikrobiyol Bul, 1976 Jul, 10(3), 307 - 11 {Providencia isolated from the stools of patients with gastroenteritis}; Berkman E; 92 providencia strains were isolated from the stools of patients with gastroenteritis . All specimens were negative for other pathogenes such as salmonella, shigella, enteropathogenic E . coli and vibrios . Viral etiology was not investigated . The distribution according to years and months is shown in the table . 17 isolations made during August, seemed rather high and gave the impression of a small epidemic. J Gen Virol, 1976 Jun, 31(3), 315 - 21 The cleavage of polyoma virus DNA by restriction enzymes KpnI and PstI; Crawford LV et al.; The action of two restriction endonucleases on polyoma virus DNA has been examined and the sites at which they cleave the DNA located . One of the enzymes, KpnI from Klebsiella pneumoniae OK8, cleaves polyoma DNA twice at about 11-6 and 59-2% from the EcoRI site . The other enzyme, PstI from Providencia stuartii 164, cleaves polyoma DNA five times at about 14-8, 16-5, 32-6, 50-3 and 80-0% from the EcoRI site . Some of the cleavages produced by these enzymes alone, or in conjunction with other endonucleases, may be of use in the isolation of regions of particular interest from the virus DNA. J Infect Dis, 1976 Mar, 133(3), 283 - 92 Reconstitution of the somatic (O-) antigenic scheme for Providencia and preparation of O-typing antisera; Penner JL et al.; The somatic (O-) antigens of the type strains of the providencia antigenic scheme were examined for their biochemical reactions and their O-specificities . The scheme of 62 O-antigens was reconstituted from 52 original type strains and 10 strains substituted for originals that either were biochemically atypical of the genus or showed inappropriate serological reactions . Thirty-six type strains showed no significant relations with other type strains, and antisera could be used for typing without absorption . Among 26 type strains, significant reciprocal relations were demonstrated, and each cross-reacting antigen was examined for specificity and for its distribution among the type strains . Antisera to these strains required absorption with cell suspensions of other type strains for production of specificity in O-typing . Each typing antiserum, at low dilution, was shown to agglutinate homologous, but not heterologous, cell suspensions of type strains, and this result demonstrated the required specificity for typing on the basis of the O-antigens. Surgery, 1976 Feb, 79(02), 224 - 8 Emergence of resistance to amikacin during treatment of burn wounds: the role of antimicrobial susceptibility testing; Overturf GD et al.; Amikacin has been used to treat Providencia stuarii infections on the Burn Service at Los Angeles County/University of Southern California Medical Center since March, 1973 . The median minimal inhibitory concentration (MIC) of strains collected on this service prior to the introduction of amikacin was 3.13 mug . per milliliter, whereas the median MIC of strains collected during the last 4 months of the study was 12.5 mug per milliliter . High bactericidal concentrations (MBC) noted at the time of initial studies predicted the emergence of resistant clones, with MBV values rising to as great as 100 mu per milliliter . Further, isolates from burn patients during the initial 5 days of treatment with amikacin had a median MIC of 6925 mug per milliliter, in contrast to values of 25 mug per milliliter in strains isolated after 5 days of treatment . The epidemiologic significance of intensive treatment of gram-negative infections occurring in a close population with selected antibiotics is discussed . The performance of susceptibility tests which included determination of bactericidal concentrations was a major tool in the recognition of the potential for selection of resistant micro-organisms. Nucleic Acids Res, 1976 Feb, 3(2), 343 - 53 The isolation and partial characterization of a new restriction endonuclease from Providencia stuartii; Smith DI et al.; We describe the isolation of a new class 2 restriction endonuclease from Providencia stuartii 164 . Using the procedure of osmotic shock treatment, we have partially purified this enzyme (Pst 1) and have begun preliminary work to characterize its specificity and requirements . Pst 1 requires Mg++ as the only cofactor and produces more than 18 cleavages in wild type lambda . We have determined the location of 7 of these cleavages by the use of deletion and insertion mutants of lambda. Zh Mikrobiol Epidemiol Immunobiol, 1975 Feb, (2), 48 - 50 {Intrageneric O-antigenic relations of Providencia bacteria}; Kholodkova EV; The authors present literature and personal data on the intragenera O-antigenic relations of the strains of a collection of bacteria belonging to Providencia genus, including standard strains of 53 serological O-groups whose antigenic interrelations were studied serologically in the tests of agglutination and adsorption of agglutinins . Experimental results confirmed the presence of unilateral relations by the O-antigen between the serological groups 01-021, 03-021 and bilateral relations between the serological groups 01-03; there were also revealed relations by the O-antigens between the serological groups 09-036, 014-023, 018-035, 039-053, and 039-027 of a unilateral character . The data obtained added information to the antigenic structure of Providencia and were of significance for the preparation of diagnostic O-sera of Providencia.
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