Cytotoxic activity in a case of adult T-cell leukemia/lymphoma with spontaneous regression

Plasmid profile analysis by agarose gel electrophoresis was performed on 42 drug resistant strains of Shigella boydii serotypes 1-5, 8, 10, 12-14, collected between 1974 and 1985 from endemic cases of shigellosis in Ethiopia, and their Escherichia coli K12 transconjugants. Resistance factors (R factors) were further characterized by incompatibility testing. Patterns of small plasmids, less than 15 kb, were similar within each of the individual S. boydii serotypes. Plasmids of about 3.3-3.7 kb were found in all strains of serotypes 2 and 4. Plasmids of about 4.3-4.6 kb were found in about 86% of strains. Serotypes 1, 2 and 3 were characterized by plasmids of about 5.6-5.7 kb. The 6.4-6.7 kb plasmid was found consistently in serotypes 1, 2, 3, 5, 8, 12 and 13 which were resistant to SSu or had an SSu resistance component in their phenotypes. Large plasmids (155-186 kb) were found in most S. boydii strains. Conjugative drug resistance plasmids, most often coding for three or less drugs, were found in about 26% of drug resistant strains. R-factors, coding for AT resistance (in types 2 and 8), and ASSuT resistance (in type 4), were compatible with all reference plasmids tested. Plasmids belonging to incompatibility groups X and N were found in serotypes 5 and 10, respectively.     

Molecular typing of Salmonella typhi strains from Dhaka (Bangladesh) and development of DNA probes identifying plasmid-encoded multidrug-resistant isolates

Seventy-eight Salmonella typhi strains isolated in 1994 and 1995 from patients living in Dhaka, Bangladesh, were subjected to phage typing, ribotyping, IS200 fingerprinting, and PCR fingerprinting. The collection displayed a high degree of genetic homogeneity, because restricted numbers of phage types and DNA fingerprints were observed. A significant number of the S. typhi strains (67%) were demonstrated to be multiple drug resistant (MDR). The vast majority of the MDR strains were resistant to chloramphenicol, ampicillin, trimethoprim, streptomycin, sulfamethoxazole, and tetracycline (R type CATmSSuT), a resistance phenotype that has also frequently been observed in India. Only two strains displayed a distinct MDR phenotype, R type AT-mSSuT. Pulsed-field gel electrophoresis demonstrated the presence of large plasmids exclusively in the MDR strains of both R types. The plasmids present in the S. typhi strains of R type CATmSSuT could be conjugated to Escherichia coli and resulted in the complete transfer of the MDR phenotype. PCR fingerprinting allowed discrimination of MDR and susceptible strains. The DNA fragments enabling discrimination of MDR and susceptible S. typhi strains by PCR were useful genetic markers for identifying MDR encoded by large plasmids of the H1 incompatibility group.     

Resistance to beta-lactam antibiotics and aminoglycosides in gram negative bacteria. 1. Molecular and genetic characterization of R-factors (author's transl)

With frequent use of aminoglycoside antimicrobials and beta-lactam antibiotics in hospitals in the last few years, the number of bacterial strains resistant to these chemotherapeutics increased. Lately, strains of E. coli, Klebsiella, Enterobacter, Serratia, Proteus and Pseudomonas resistant to many antimicrobials (ampicillin, carbenicillin, cephalothin, chloramphenicol, gentamycin, tobramycin, sisomycin, neomycin, paromomycin, kanamycin, streptomycin, spectinomycin, tetracycline, sulphonamides) were isolated from patients of the university hospital in Zuerich. The resistant phenotype of two representative strains (Klebsiella pneumoniae 1 and Serratia marcescens 2) could be transferred by mixed cultivation to E. coli K-12. Multiple resistance of strain 1, and addition, could be transferred to Salmonella typhimurium, Serratia marcescens, Providencia, Proteus mirabilis and Klebsiella pneumoniae in varying frequencies. Transfer to Pseudomonas aeruginosa, however, could not be achieved. Spontaneous instability of resistance was observed in 0.15% of the cells of an overnight brothe culture and in 90% of the cells of a three months old culture. Conjugation, instability and the response to the sex phages MS-2 and If-1 suggested that resistance was mediated by a monomolecular R-factor, belonging to the fi+-type. This suggestion was confirmed by molecular characterization of the resistance plasmids. After transfer of the R-factors of K. pneumoniae 1 (R-FK 1) and Serratia marcescens 2 (R-FK2) into E. coli K-12, plasmid DNA was labelled with (methyl-3H) thymidine, and isolated by isopycnic centrifugation in cesiumchlorid-ethidium-bromide. Analysis of plasmid DNA then was carried out by sedimentation in a 5-20% neutral sucrose gradient together with reference plasmids of known molecular weights and sedimentation constants. The analysis revealed that R-FK1 had a molecular weight of 54 X 10(6) and R-FK2 of 50 X 10(6) daltons. The values were confirmed by contour length measurements of open circular forms with an electron microscope. A comparison of the sedimentation profile of labelled plasmid DNA from strain 1 and 14C-labelled DNA of E. coli K-12 (R-FK1) showed that the wild-type strain contained, besides the large resistance plasmid, at least two smaller "cryptic" plasmids. These smaller plasmid molecules were also found in antibiotic susceptible variants of strain 1, which did not contain the 54 X 10(6) dalton plasmid molecule, responsible for the resistant phenotype. The number of copies of R-FK1 in E. coli K-12 was determined to be 2, indicating stringent control of replication. It is discussed that the growing number of isolations of strains of Escherichia, Klebsiella, Serratia, Proteus, Providencia and Pseudomonas, exhibiting the same resistance phenotype, results from the spread of the R-factor described above among the hospital bacterial flora.     

A male child with the rumpshaker mutation, X-linked spastic paraplegia/Pelizaeus-Merzbacher disease and lysinuria

Of 52 antibiotic-resistant Bordetella bronchiseptica isolates from cats, ten carried plasmids. Only two of these plasmids, pLV1400 and pLV1401, were self-transmissible to Escherichia coli K12; both plasmids encoded resistance to ampicillin, tetracycline, sulphonamides, streptomycin and mercuric chloride, and were of incompatibility group P (IncP). Transferable tetracycline resistance has not been reported in B. bronchiseptica previously. The plasmids were identical in size (c.51 kb), restriction endonuclease digestion pattern and gene sequences (trfA and korA) within the IncP replicon. The trfA and korA sequences differed from those of the archetypal IncP plasmids RP4 and R751. Although the two B. bronchiseptica isolates were from epidemiologically and geographically separated cats, pulsed-field gel electrophoresis of their XbaI- or DraI-digested chromosomal DNA indicated that they were genotypically identical. The plasmid-encoded ampicillin resistance was mediated by a penicillinase of molecular weight 49,000, and pI 8.45 which was inhibited by clavulanate (IC50 = 0.1 mg/L) and tazobactam (IC50 = 0.42 mg/L) but not by parachloromercuribenzoate or EDTA. The high-level tetracycline resistance was mediated by a class C efflux mechanism that has not been described previously in this genus. The presence of transferable multi-drug resistance on a promiscuous plasmid may limit options for therapy of respiratory tract infection in companion and farm animals.     

The molecular basis of partial penetrance of splicing mutations in cystic fibrosis

Biotin-labelled DNA probes and restriction-endonuclease digestion (RED) with HindIII were used to study the diversity of resistance plasmids (R-plasmids) from 414 Escherichia coli isolates: 168 from children living in close contact with antibiotic-fed poultry and 246 from the chickens. Full sensitivity to all 10 antimicrobials tested was more common in the isolates from poultry than in those from the children (36.2% v. 9.5%; P < 0.001). Multi-drug resistance, to at least two of the antimicrobials, was relatively common in the isolates from the children (85.5% v. 26.00%; P < 0.001). Overall, 31% of the poultry isolates were resistant to tetracycline alone. Resistance to amoxycillin was due to production of TEM-1 (89%) and TEM-2 (11%). In > 71% of the isolates from children and 79% of those from poultry, resistance was encoded on a 100-110-kb transferable plasmid belonging to incompatibility group FII. However, RED patterns of R-plasmids from the two groups of isolates were highly diverse and not indicative of any close relatedness. This difference in patterns and in the levels of multi-drug resistance indicate that the isolates from the children and those from the poultry represent two distinct pools of resistance plasmids.     

Plasmid-mediated gentamicin resistance of Pseudomonas aeruginosa and its lack of expression in Escherichia coli

We isolated 11 nonconjugative plasmids mediating resistance to aminoglycoside antibiotics, including gentamicin, from Pseudomonas aeruginosa strains. Their genetic properties were investigated in both P. aeruginosa and Escherichia coli transformants. The plasmid molecular weights ranged from 11 x 10(6) to 24 x 10(6). A low level or complete absence of gentamicin resistance was observed when these plasmids were introduced into E. coli, but gentamicin resistance was restored when the plasmids were transferred back to P. aeruginosa from E. coli. Aminoglycoside-modifying enzyme activity was detected in P. aeruginosa harboring these plasmids, but was absent or greatly reduced in E. coli strains. This lack of expression may explain the observed decrease in aminoglycoside resistance.     

Anomalous patterns of callosal connections develop in visual cortex of monocularly enucleated hamsters

Six multiply resistant isolates of Salmonella typhimurium var. copenhagen with high-level resistance to fluoroquinolones (e.g., MIC of ciprofloxacin: 32 micrograms/ml) were isolated from human patients (n = 3) and from cattle (n = 3). The isolates were examined by complementation tests using a set of broad-host-range plasmids, which carry either the gyrA+ or the gyrB+ genes or a combination of both from Escherichia coli K-12. The results indicated a combination of gyrA and gyrB mutations in all isolates. Subsequent direct sequencing of PCR-generated internal DNA fragments of gyrA revealed an identical double mutation in all six isolates (Ser-83-->Ala and Asp-87-->Asn). In addition, the results of phenotypic (i.e., phagetype, biotype, serotype) and genotypic characterization [i.e., ribotyping and polymerase chain reaction fingerprinting (PCR-fingerprinting)] were identical for all six isolates and were distinguishable from a quinolone-susceptible strain of the same serovar and an unrelated isolate of S. typhimurium. These data indicate the clonal identity of the fluoroquinolone-resistant strains of S. typhimurium isolated from men and cattle in Germany.     

pRJ5: a naturally occurring Staphylococcus aureus plasmid expressing constitutive macrolide-lincosamide-streptogramin B resistance contains a tandem duplication in the leader region of the ermC gene

The 2.55 kb Staphylococcus aureus plasmid, pRJ5, confers constitutive resistance to macrolide-lincosamide-streptogramin B (MLS) antibiotics. pRJ5 is nearly identical to the inducible MLS resistance plasmid pT48, and has homology with the S. aureus plasmids pE194 and pSN2. The HindIII-C and/or Hind-B fragments were required for stable maintenance of the plasmid and probably carry palA. Plasmids pRJ5 and pT48 were shown to belong to the same incompatibility group, Inc12 (L). DNA sequencing showed that pRJ5 contains a 28 bp direct tandem duplication in the leader/attenuator region of ermC. This is likely to change the secondary structure of the methylase mRNA, allowing constitutive expression of ermC. The type of mutation found on plasmid pRJ5 is different from those observed in similar 2.5 kb constitutive MLS-resistance plasmids isolated from other Gram-positive bacteria, including staphylococci.     

Antimicrobial resistance and epidemiological study of Haemophilus influenzae strains isolated in Portugal. The Multicentre Study Group

In the course of a multicentric surveillance study, nine laboratories sent 375 isolates of Haemophilus influenzae to the Sector de Resistência aos Antibióticos (SRA) from the National Institute of Health in Lisbon, between 1 January and 31 December 1992. The majority of the H. influenzae isolates were from the respiratory tract (84.8%); only 5.1% were of invasive origin. Overall resistance for ampicillin was 11.7%, tetracycline 3.7%, and chloramphenicol 2.4%. All isolates tested were fully susceptible to cefotaxime, ceftriaxone, ciprofloxacin and rifampicin. Multiresistance was rare, occurring only in 2.4% of the isolates, although 50% of the ampicillin resistant strains had at least one additional resistance marker. Forty two isolates (11.2%) produced a TEM-1 type beta-lactamase, as shown by isoelectric focusing. beta-lactamase production was not detected in two of the ampicillin resistant strains. Fifteen of the 42 beta-lactamase producing strains (35.7%) contained detectable DNA plasmid: nine harboured large plasmids with an apparent molecular mass of 45 or 54 kb depending on their resistance phenotype and six harboured a small plasmid of 5 kb. In order to study transfer of resistance in both ampicillin and multiresistant strains conjugation experiments were performed for 14 isolates, seven of which harboured a large plasmid and seven had no detectable plasmid DNA. All 14 transferred their resistance phenotype but only a single large plasmid could be demonstrated in ten transconjugants. Restriction endonuclease analysis of plasmids from six representative transconjugants, isolated in different hospitals, revealed that there was no dissemination of a single R plasmid, which suggests an independent process of acquisition of resistance genes.     

Ciprofloxacin resistance in gram negative bacilli. Epidemiologic aspects

BACKGROUND: The aim of the present was to study the resistance to ciprofloxacin (CIP) in gram negative bacilli (April 1990-March 1992) and to determine the temporary distribution of the resistant strains by species, samples and departments. METHODS: Seven thousand four hundred seventy-eight samples were studied. The identification and determination of the sensitivity was performed by the PASCO (Difco) system. Haemophilus spp. and Campylobacter spp. were excluded from the study. The microorganisms with CIM 2 mg/l were considered as resistant (CIPr). RESULTS: Four hundred eighty-one CIPr isolations were identified (6.4%). With regard to the percentage of resistant strains, the species with the highest were: Providencia stuartii (50%); Pseudomonas cepacia (44.4%); Xanthomonas maltophilia (26.9%); Acinetobacter baumannii (25.8%); Pseudomonas aeruginosa (16%); Citrobacter freundii (12.5%); Serratia marcescens (8.4%); Enterobacter cloacae (5.8%) and Escherichia coli (4.3%). With respect to the absolute number of resistant strains, the most frequent resistant strains were: P. aeruginosa (205), E. coli (144), and A. baumannii (41). Isolation of E. coli and A. baumannii CIPr increased over the study period. Forty-four point five percent of the E. coli CIPr were of extrahospitalary origin; most of the A. baumannii (92.9%) and P. aeruginosa (77.6%) in contrast were of intrahospitalary origin. CONCLUSIONS: P. aeruginosa, E. coli, and A. baumannii are the most frequently resistant species. The frequency of isolation of resistant strains of E. coli and A. baumannii significantly increased (p < 0.001) over the two years of the study.     

Micronucleus assays using cytochalasin-blocked MCL-5 cells, a proprietary human cell line expressing five human cytochromes P-450 and microsomal epoxide hydrolase

A total of 100 S. hyicus strains isolated from healthy piglets and piglets with exudative epidermitis originating from 100 different herds was examined for drug-resistance and prevalence of plasmids. Resistance to macrolide/linosamide antibiotics could be related to plasmids in 55 (93%) of the 59 resistant strains: A plasmid of 2.4 kb mediating resistance to macrolides and lincosamides was observed in 25 strains, and a plasmid of 11.5 kb mediating resistance to both macrolides/lincosamides and tetracycline was observed in 30 strains. A plasmid with a molecular weight of 4.5 kb was shown by curing experiments to be associated with resistance to tetracycline in 12 strains. All together, 47 strains were resistant to tetracycline. In 42 (89%) of these strains tetracycline-resistance was found to be encoded by plasmids. Fifty six strains were resistant to streptomycin, and resistance was associated with the presence of a 4.4 kb plasmid in 17 strains studied. Resistance to penicillin, observed in 44 strains, and resistance to kanamycin, observed in 15 strains, could not be related to plasmids in any of these strains. The 11.5 kb plasmid was observed in 39% of the strains isolated from piglets with EE, and in 7% of the strains isolated from healthy piglets. Despite its higher prevalence in strains from piglets with EE, the 11.5 kb plasmid could not be shown to encode production of capsule or exfoliative substances: factors which might play a role in the development of exudative epidermitis in piglets.     

Variations in hospitalization rates for asthma among black and minority ethnic communities

The European Suspension Test was used to assess the relative resistance of 19 individual Listeria monocytogenes genotypes, isolated from the poultry processing environment, to three commercially used disinfectants employed in the plant at the time of their isolation. To establish the relative resistance between the strains, the concentration of each disinfectant was reduced until inter-strain variation became apparent. For Darasan 214 and 7058, variation was detected at 0.1% and 0.5% v/v, respectively, while Daraclean 7361 had to be reduced to only 2.5% v/v. At these concentrations, the mean microbiocidal effect (ME) of each disinfectant ranged between 4.3 and 3.1 log10 reduction in cfu ml-1. Significant differences between the strains were obtained with respect to their resistance to the disinfectants employed (P < 0.01), but the overall log10 reduction for genotypes 'A1' and 'A2', which were found to persist in the poultry processing environment, were not found to be significantly different from the genotypes which had been isolated on a more sporadic basis (P > 0.05). The L. monocytogenes strains fell into four groups with respect to incidence and size of plasmids isolated. The first group contained strains which carried two plasmids (5 and 40 MDa) and the other three (groups 2, 3 and 4) comprised strains which carried a single plasmid (14, 47 and 52 MDa, respectively). There was no correlation between persistent and sporadic strains with respect to incidence and size of plasmids isolated. Moreover, the strains which carried no plasmids were found to be as resistant to the disinfectants as those which did carry plasmids, suggesting that the plasmids isolated did not confer resistance of L. monocytogenes planktonic cells to the disinfectants tested. Therefore, it is unlikely that the strains which had been found to persist in the poultry processing environment did so by means of plasmid-mediated resistance to the commercial disinfectants used.     

Post-transfusion hyperhaemolysis in a patient with sickle cell disease: use of steroids and intravenous immunoglobulin to prevent further red cell destruction

Forty nine multiple drug resistant strains of E. coli isolated from UTI were serotyped. The pattern was found to be 057 (eight strains); 0109 (four strains); 020, 038, 068, 0106, 0148. Rough (three each). 012, 054, 0101, 0160 (two each) and 02, 032, 046, 053, 060, 065, 090, 091, 0117, 0118, untypable (one each). The resistance pattern of all E. coli were identified and matted with recepient strain in penassay broth and in human urine. In a penassay broth transfer of resistance was demonstrated in 38 strains (77.5%) while in human urine transfer was demonstrated only in 14 strains (28.57%).     

Characterization of plasmids which mediate resistance to multiple antibiotics in gram-negative bacteria of nosocomial origin

BACKGROUND: The genetic and molecular mechanisms involved in antimicrobial resistance of 10 strains of gramnegative bacilli (1 Serratia marcescens; 2 Escherichia coli; 1 Proteus mirabilis; 4 Klebsiella pneumoniae; 1 Enterobacter cloacae y 1 Alcaligenes faecalis), isolated from adult patients with nosocomial pulmonary infection at the in-patient facilities of the University Hospital of Los Andes, Mérida, Venezuela, have been studied. METHODS: The antimicrobial susceptibility was determined by minimum inhibitory concentrations using the dilution method in agar. The study of extrachromosomal genes was carried out by conjugation, bacterial infection with the bacteriophage M13 and curing of plasmid by acridine orange. The plasmids were isolated by alkaline lysis and analysis of restriction endonuclease digestion was carried out separately using the enzymes EcoRI and HindIII. A DNA probe, derived from the region which encodes the TEM-1 beta-lactamase of the plasmid pBR322 was used for dot-blot hybridization tests. RESULTS: All of the gramnegative bacilli showed resistance to ampicillin, carbenicillin and cephalothin (> 128 micrograms/ml) and 3 strains also showed resistance to gentamicin (> 64 micrograms/ml). Genetic and molecular procedures showed the presence of conjugative plasmids of approximately 54 kb in all the 10 strains. The restriction patterns obtained by using EcoRI and HindIII indicated common DNA fragments in most of the plasmids studied. The dot-blot hybridization tests confirmed homology between the plasmids and the DNA probe used (TEM-1 beta-lactamase). CONCLUSIONS: In this study, the gramnegative bacteria of nosocomial origin harbored self-transferable plasmids of approximately 54 kb, which mediate resistance to gentamicin and encode a beta-lactamase of the TEM group.     

An analysis of excessive running in the development of activity anorexia

Several plasmids that were isolated from complement-resistant Pasteurella multocida or Escherichia coli were evaluated for phenotypic markers. Plasmid p2267, isolated from a tetracycline-resistant, complement-resistant fowl cholera field isolate of P. multocida (PM2267), was used to transform a K-12 E. coli (C600); this resulted in increased complement resistance, which was eliminated by curing. Either of two plasmids (p1870 or p70-1, isolated from P. multocida and E. coli, respectively) conferred an increase in complement resistance and invasiveness to turkey epithelial cells when expressed in the Clemson University (CU) vaccine strain of P. multocida. Additionally, the CU strain containing p1870 was more virulent in turkey challenge, and the plasmid appeared amplified in vivo. No detectable differences in major outer-membrane proteins, capsule, or carbohydrate fermentation were found to be associated with the acquisition of these plasmids.     

Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies

Within the species Escherichia coli, there are commensal strains and a variety of pathogenic strains, including enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and urinary tract infection (UTI) strains. The pathogenic strains are identified by serotype and by possession of specific virulence determinants (toxins and adhesions, etc.) encoded by either monocistronic genes, plasmids, or pathogenicity islands. Although there are studies on the relationships between selected pathogenic strains, the relatedness among the majority of the pathogenic forms to each other, to commensal E. coli, and to the genus Shigella (which has often been suggested to be part of E. coli) has not been determined. We used multilocus enzyme electrophoresis (MLEE) at 10 enzyme loci and the sequence of the mdh housekeeping gene to study the genetic relationships of pathogenic E. coli strains (including Shigella clones), namely, 5 EPEC strains (serotypes O111 and O55), 3 EHEC strains (serotype O157), 6 ETEC strains (serotypes O78, O159, and O148), 5 EIEC strains (serotypes O124, O28, and O112), and 13 Shigella strains representing clones Flexneri, Dysenteriae, Boydii, and Sonnei, to commensal E. coli strains. Both the MLEE and mdh sequence trees reveal that EPEC, EHEC, ETEC, EIEC, and UTI strains are distributed among the ECOR set groups, with no overall clustering of EPEC, ETEC, EIEC, or UTI strains. The genus Shigella is shown to comprise a group of closely related pathogenic E. coli strains. Six pathogenic strains, i.e., M502 (EIEC; O112ac:NM), M503 (EPEC; O111:H12), M526 (ETEC; O159:H4), M522 (EPEC; O111ac:H12), M524 (ETEC; O78:H11), and M506 (ETEC; O78:H11), were found to have mdh sequences identical to those of five ECOR group A strains (ECOR5, ECOR10, ECOR14, ECOR6, and K-12). All 11 strains are closely related by MLEE. The results indicate that pathogenic strains of E. coli do not have a single evolutionary origin within E. coli but have arisen many times. The results also suggest the possibility that any E. coli strain acquiring the appropriate virulence factors may give rise to a pathogenic form.     

Plasmids isolated from marine sediment microbial communities contain replication and incompatibility regions unrelated to those of known plasmid groups

Two hundred ninety-seven bacteria carrying plasmids that range in size from 5 to 250 kb were identified from more than 1,000 aerobic heterotrophic bacteria isolated from coastal California marine sediments. While some isolates contained numerous (three to five) small (5- to 10-kb) plasmids, the majority of the natural isolates typically contained one large (40- to 100-kb) plasmid. By the method of plasmid isolation used in this study, the frequency of plasmid incidence ranged from 24 to 28% depending on the samples examined. Diversity of the plasmids occurring in the marine sediment bacterial populations was examined at the molecular level by hybridization with 14 different DNA probes specific for the incompatibility and replication (inc/rep) regions of a number of well-characterized plasmid incompatibility groups (repB/O, FIA, FII, FIB, HI1, HI2, I1, L/M, X, N, P, Q, W, and U). Interestingly, we found no DNA homology between the plasmids isolated from the culturable bacterial population of marine sediments and the replicon probes specific for numerous incompatibility groups developed by Couturier et al. (M. F. Couturier, F. Bex, P. L. Bergquist, and W. K. Maas, Microbiol. Rev. 52:375-395, 1988). Our findings suggest that plasmids in marine sediment microbial communities contain novel, as-yet-uncharacterized, incompatibility and replication regions and that the present replicon typing system, based primarily on plasmids derived from clinical isolates, may not be representative of the plasmid diversity occurring in some marine environments. Since the vast majority of marine bacteria are not culturable under laboratory conditions, we also screened microbial community DNA for the presence of broad- and narrow-host-range plasmid replication sequences. Although the replication origin of the conjugally promiscuous broad-host-range plasmid RK2 (incP) was not detectable in any of the plasmid-containing culturable marine isolates, DNA extracted from the microbial community and amplified by PCR yielded a positive signal for RK2 oriV replication sequences. The strength of the signal suggests the presence of a low level of the incP replicon within the marine microbial community. In contrast, replication sequences specific for the narrow-host-range plasmid F were not detectable in DNA extracted from marine sediment microbial communities. With the possible exception of mercuric chloride, phenotypic analysis of the 297 plasmid-bearing isolates did not demonstrate a correlation between plasmid content and antibiotic or heavy metal resistance traits.     

A fundamental study of 99mTc-HMPAO double injection method and clinical application to stress scintigraphy in orthostatic hypotension

Enterococcus faecium, which was highly resistant to vancomycin (MIC 256 mg/liter), but susceptible to teicoplanin (MIC 2 mg/liter), caused two distinct episodes of infection on a renal unit in the United Kingdom. Pulsed field gel electrophoresis (PFGE) indicated that a single strain caused the first episode, while the second episode, which occurred 1 year later, involved multiple strains, all of which were distinct from the original strain. Vancomycin resistance in all but one of these strains was mediated by transferable plasmids that carried the vanB glycopeptide resistance gene. Transfer either of resistance plasmids or the vanB resistance determinant itself to different strains occurred during the second episode. Plasmid-mediated vanB resistance has not been widely documented. A retrospective study of a reference collection revealed two other vanB-encoding plasmids from an E. faecalis and an E. faecium referred from two further UK centers. Although restriction analysis indicated no similarity between the plasmids from the three different centers, all contained a 2.1-kb EcoRV fragment that hybridized with a probe for the vanB gene. This suggests that there has been dissemination of a conserved glycopeptide resistance determinant, of which vanB is a part.     

Pneumococcal resistance to antimicrobial agents in the province of Québec, Canada

The serogroup/serotypes (SGTs) and antimicrobial susceptibilities to 10 antimicrobial agents of 110 clinical strains of Streptococcus pneumoniae were determined. Strains intermediately resistant or highly resistant to penicillin G (80 of 110) belonged predominantly to SGTs 23 (45.0%), 19 (13.7%), 6 (10.0%), 9 (6.2%), and 14 (3.7%). The MICs of all cephalosporins, tetracycline, trimethoprim-sulfamethoxazole, and chloramphenicol increased along with the MICs of penicillin G. However, erythromycin resistance and clindamycin resistance were observed more frequently among the intermediately penicillin-resistant strains. Multiple resistance was observed for 32 strains, of which 25 were highly resistant to penicillin G and belong to SGT 23F. All strains were susceptible to vancomycin.     

Inhibition of bacteriophage lambda, T1, and T7 development by R plasmids of the H incompatibility group

R plasmids from chloramphenicol-resistant salmonella from Ontario are shown to belong to the H(2) incompatibility subgroup and to mediate a broad-spectrum, phage inhibition function.