Enterobacter plasmids: molecular epidemiology.
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DNA, Bacterial↗
Biomedical subjects
J F John
Publications and source records attributed to J F John.
In vivo R-plasmid transfer in a patient with a mixed infection of shigella dysentery.
Transfer of shigella R-plasmids in vivo has seldom been demonstrated. Strains of Shigella dysenteriae type 1 and Shigella flexneri type 5b were isolated from a Bulgarian traveller who visited Vietnam and developed dysentery, which was treated with trimethoprim/sulfamethoxazole (TMP/SMZ) for a short time. Both species of shigellae are unusual in Bulgaria where strains of S. sonnei predominate. Both shigella strains were multiresistant to the same antimicrobial agents. Each strain contained a 48-kilobase plasmid that conferred the entire resistance phenotype to a susceptible Escherichia coli. Restriction endonuclease patterns of plasmid DNA from the respective strains were identical. Transmissible plasmids of the same resistance phenotypes and restriction patterns were isolated from the patient's colonic E. coli. Transconjugants hybridized to a dihydrofolate reductase type I-DNA probe. These studies support the hypothesis that R-plasmid transfer may occur between non-pathogenic, faecal strains and pathogenic shigellae, a process that may have been facilitated by inadequate treatment with TMP/SMZ at the onset of the illness.
Aged↗
Radiographic determination of the anatomic hip joint center. A cadaver study.
We studied 31 human pelvic cadaver specimens with 57 intact hip joints. The teardrop, which represents the inferior portion of the acetabular notch, was identified as also was the center of rotation in the articulating femoral head. The pelvic height and the horizontal and vertical distances between the acetabular notch and the hip joint center were measured. We found the center of rotation of the hip joint to be 13 percent of the pelvic height lateral and 7 percent of the pelvic height superior to the teardrop. No sex difference was found.
Aged↗
Respiratory syncytial virus infection in a cardiac surgery intensive care unit.
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Aged↗
Methicillin-resistant Staphylococcus aureus: a consensus review of the microbiology, pathogenesis, and epidemiology with implications for prevention and management.
Methicillin-resistant Staphylococcus aureus (MRSA) has become a major nosocomial pathogen in community hospitals, long-term-care facilities, and tertiary care hospitals. The basic mechanism of resistance is alteration in penicillin-binding proteins of the organism. Methods for isolation by culture and typing of the organism are reviewed. MRSA colonization precedes infection. A major reservoir is the anterior nares. MRSA is usually introduced into an institution by a colonized or infected patient or health care worker. The principal mode of transmission is via the transiently colonized hands of hospital personnel. Indications for antibiotic therapy for eradication of colonization and treatment of infection are reviewed. Infection control guidelines and discharge policy are presented in detail for acute-care hospitals, intensive care and burn units, outpatient settings, and long-term-care facilities. Recommendations for handling an outbreak, surveillance, and culturing of patients are presented based on the known epidemiology.
Anti-Bacterial Agents↗
Increased frequency of large R-plasmids in Klebsiella pneumoniae colonizing patients with spinal cord injury.
From 1978 to 1988 strains of gentamicin-susceptible (Gms) and gentamicin-resistant (Gmr) Klebsiella pneumoniae were saved from annual surveillance cultures of the perineal region of patients with spinal cord injury (SCI). Of 38 strains selected for further study (24 Gms and 14 Gmr), there were 23 different serotypes (two nontypable). Fourteen Gms as well as 14 Gmr strains displayed no common plasmid patterns, but all contained a large plasmid of 168-208 kb. Among the 14 Gmr strains, nine had large conjugative plasmids of approximately the same size (166-193 kb), which conferred to a susceptible Escherichia coli host an identical resistance pattern: ampicillin, chloramphenicol, gentamicin, piperacillin, trimethoprim-sulfamethoxazole, tetracycline, and tobramycin. Of the nine transconjugants, eight contained a single plasmid. One transconjugant contained a 168- and 80-kb plasmid. Restriction endonuclease digestion patterns of the R-plasmids revealed minimal similarity. We conclude that, during a 10-year period, different large R-plasmids have spread among multiple serotypes of K. pneumoniae in spinal cord injury (SCI) patients in one rehabilitation hospital. We hypothesize that other genes located on large, R-, and non-R-plasmids may confer an additional advantage for colonization by K. pneumoniae in SCI patients.
Conjugation, Genetic↗
Widespread colonization of personnel at a Veterans Affairs medical center by methicillin-resistant, coagulase-negative Staphylococcus.
A serial prospective survey of nasal colonization of hospital personnel by methicillin-resistant coagulase-negative staphylococci (MRCNS) was conducted at a Veterans Affairs medical center on three occasions over a 16-month period. The epidemiological typing systems used to assess relatedness included antimicrobial susceptibility profiles; biotyping; phage typing; plasmid profiles; restriction fragment length polymorphism (RFLP) analysis with ribosomal RNA; and plasmid hybridization with a 1.68-MD plasmid as the DNA probe. Forty-three percent of all personnel and 62% of all nurses were colonized with MRCNS. Nurses on the wards (72%) and in the intensive care unit (73%) were significantly more likely to be colonized with MRCNS than nurses who had less contact with patients or those who worked in the operating room. The molecular epidemiological typing systems indicated some degree of relatedness among the strains. Specifically, riboprobe analysis revealed a Dice coefficient of > 90%. However, each typing system detected dissimilarity among strains. Further studies are needed to determine the role that such human reservoirs of MRCNS serve in horizontal transmission to and subsequent infection of hospitalized patients.
Bacteriophage Typing↗
Randomized double-blinded trial of rifampin with either novobiocin or trimethoprim-sulfamethoxazole against methicillin-resistant Staphylococcus aureus colonization: prevention of antimicrobial resistance and effect of host factors on outcome.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen in hospitals. Current antimicrobial regimens for eradicating colonizing strains are not well defined and are often complicated by the emergence of resistance. The combination of novobiocin plus rifampin in vitro and in vivo was found to prevent the emergence of resistant populations of initially susceptible strains of MRSA, particularly resistance to rifampin. We therefore studied, in a randomized, double-blind, multicenter comparative trial, the combination of novobiocin plus rifampin versus trimethoprim-sulfamethoxazole (T/S) plus rifampin in order to determine the efficacy of each regimen in eradicating MRSA colonization and to further characterize the host factors involved in the response to this antimicrobial therapy. Among the 126 individuals enrolled in the study, 94 (80 patients; 14 hospital personnel) were evaluable. Among the 94 evaluable subjects, no significant demographic or medical differences existed between the two treatment groups. Successful clearance of the colonizing MRSA strains was achieved in 30 of 45 (67%) subjects receiving novobiocin plus rifampin, whereas successful clearance was achieved in 26 of 49 (53%) subjects treated with T/S plus rifampin (P = 0.18). The emergence of resistance to rifampin developed more frequently in 14% (7 of 49) of subjects treated with T/S plus rifampin than in 2% (1 of 45) of subjects treated with novobiocin plus rifampin (P = 0.04). Restriction endonuclease studies of large plasmid DNA demonstrated that the same strain was present at pretherapy and posttherapy in most refractory cases (24 of 29 [83%] subjects). Among the 56 successfully treated subjects, clearance of MRSA was age dependent: 29 of 36 (80%) subjects in the 18- to 49-year-old age group, 19 of 35 (54%) subjects in the 50- to 69-year-old age group, and 8 of 23 (35%) in the 70- to 94-year-old age group (P < 0.01). Clearance was also site dependent; culture-positive samples from wounds were related to a successful outcome in only 22 (48%) of 46 subjects, whereas culture-positive samples from sites other than wounds (e.g., nares, rectum, and sputum) were associated with a success rate of 34 of 48 (71%) subjects (P = 0.02). Foreign bodies in wounds did not prevent the eradication of MRSA by either regimen. T/S plus rifampin was less effective in clearing both pressure and other wounds, whereas novobiocin plus rifampin was equally effective in clearing both pressure and other wounds. There were no significant differences in toxicity between the two regimens. Thus, the combination of novobiocin plus rifampin, in comparison with T/S plus rifampin, was more effective in preventing the emergence of resistance to rifampin and demonstrated a trend toward greater activity in clearing the MRSA carrier state. The response to either combination depended on host factors, particularly age and the site of MRSA colonization.
Adolescent↗
Relatedness of strains of methicillin-resistant coagulase-negative Staphylococcus colonizing hospital personnel and producing bacteremias in a neonatal intensive care unit.
The emergence of methicillin-resistant coagulase-negative Staphylococcus as a major bacterial pathogen in neonatal intensive care units has stimulated interest in the epidemiology of spread of the organism. During a 12-month "epidemic" of bacteremias with methicillin-resistant coagulase-negative Staphylococcus we compared the characteristics of bacteremic and personnel nasally-carried strains by traditional and biomolecular methods. Sixty-two percent of neonatal intensive care unit nurses were colonized with methicillin-resistant coagulase-negative Staphylococcus with similar speciation to bacteremic strains. Inspection of plasmid profiles revealed a moderate degree of similarity between bacteremic and colonizing strains although genomic DNA restriction patterns showed diversity. Ribotype patterns were highly conserved (90%) in personnel strains. A 2.6-kilobase plasmid DNA probe hybridized to similarly sized plasmids and larger plasmids in one-half of the strains. We hypothesize that related methicillin-resistant strains may be transferred among personnel and neonates in the neonatal intensive care unit. Epidemiologic studies of coagulase-negative staphylococci should consider multiple molecular techniques to relate strains.
Bacteremia↗
Activities of newer fluoroquinolones against Shigella sonnei.
The activities of six fluoroquinolones were determined for 117 separate strains of Shigella sonnei. The order of increasing activity (MICs for 90% of strains tested) was enoxacin (0.25 micrograms/ml), temafloxacin (0.032 micrograms/ml), sparfloxacin (0.016 micrograms/ml), CI-960 (0.008 micrograms/ml), ciprofloxacin (0.008 micrograms/ml), and PD-131628-2 (0.008 micrograms/ml). These data, along with results of killing and mutational rate studies, showed that all six fluoroquinolones were highly inhibitory against S. sonnei and five fluoroquinolones were rapidly and persistently bactericidal.
Anti-Infective Agents↗
Molecular epidemiology of trimethoprim-resistant Shigella boydii serotype 2 strains from Bulgaria.
In 1990 an increased number of strains of Shigella boydii serotype 2 were isolated from different regions of Bulgaria. Strains were reported as sporadic, although they showed identical phenotypic characteristics, including resistance to ampicillin, carbenicillin, streptomycin, sulfonamide, tetracycline, ticarcillin, and trimethoprim. The objective of this study was to determine the genetic relatedness of the strains and the mechanism of their antimicrobial resistance. Plasmid fingerprinting showed an identical pattern for 23 of 25 of the selected strains. All 25 strains tested transferred their resistances en bloc to an Escherichia coli recipient. Transconjugants contained a 112-kb R plasmid which carried all the resistance genes, including that conferring type I dihydrofolate reductase-mediated trimethoprim resistance (MIC greater than 2,000 micrograms/ml). Riboprobe analysis showed identical restriction length fragment polymorphisms, suggesting a highly conserved genome. All findings indicate that strains of S. boydii serotype 2 isolated in 1990 from different regions of Bulgaria were highly related genetically and can be considered representatives of a single bacterial clone. The presence of an R plasmid and selection pressure because of the usage of antimicrobial agents, particularly trimethoprim, have likely facilitated the spread of the clone throughout the country.
Bulgaria↗
Antibiotic resistance in long-term care facilities.
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Aged↗
Bactericidal action of modulated ultraviolet light on six groups of Salmonella.
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Colony Count, Microbial↗
Isomeric DNA ladder formation of a plasmid encoding tobramycin resistance from Escherichia coli.
During a 20-month survey of resistance to three aminoglycosides (gentamicin, tobramycin, and amikacin) in Escherichia coli at a university hospital, six tobramycin-, kanamycin-resistant isolates containing a 50 kilobase conjugative R-plasmid which encoded an aminoglycoside phosphotransferase (APH-(3')) were isolated. The APH-(3') conferred resistance to kanamycin (MIC = 100 mg/L) but not to tobramycin (MIC = 20 mg/L). In both the original isolates and transconjugants the six R-plasmids demonstrated an isomeric ladder in the range of 50-112 kb, which was enhanced by exposure of the bacterial cultures to tobramycin. pJFJ2522 is the prototype for this group of plasmids. Bacterial DNA gyrase reversed the isomeric DNA ladder in pJFJ25222 by increasing the supercoiling of the plasmid DNA. Regardless of the level of supercoiling, the plasmids produced indistinguishable restriction endonuclease fragment patterns. The clinical isolates containing these plasmids demonstrated different restriction fragment length polymorphism (RFLP) of their EcoRI digested genomic DNA using E. coli rRNA as a probe. Ladder formation was plasmid specific since other tobramycin R-plasmids did not form a ladder, but it was not host specific. pJFJ25222 formed a ladder in a recA- host and displayed the same restriction pattern in a recA- as in a recA+ environment. In conclusion, pJFJ2522 contains a new tobramycin resistance gene whose mechanism of resistance is not known and whose product probably influences the isomerization of the plasmid.
Conjugation, Genetic↗
Methicillin-resistant Staphylococcus aureus outbreak at a Veterans' Affairs Medical Center: importance of carriage of the organism by hospital personnel.
The reported prevalence of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) by hospital personnel averages 2.5%. From August 1985 to September 1987, 155 patients at our Veterans' Affairs Medical Center (VAMC) were colonized or infected with MRSA. In December 1986, only two (2.1%) of 94 healthcare workers were identified as nasal carriers. Prompted by a sharp increase in the number of patients with MRSA in early 1987, contact tracing identified 450 employees, of whom 36 (8%) were nasal carriers. Thirty-five percent of surgical residents (7 of 20) were nasal carriers. Prior to being identified as a nasal carrier, one surgical resident was associated with the inter-hospital spread of the VAMC MRSA strain to the burn unit of the affiliated university hospital. Three family members of two employee carriers were also found to harbor the epidemic strain. All 36 carriers were decolonized with various antimicrobial combinations. Vigorous infection control measures were effective in controlling the epidemic. The frequency of MRSA carriage by hospital personnel at our medical center during the epidemic proved higher than previously appreciated. Thus, healthcare workers may comprise a sizable MRSA reservoir. During an MRSA epidemic, infection control should attempt to identify and decolonize this hospital reservoir, as these individuals can disseminate MRSA both within the hospital as well as into the community.
Cross Infection↗
Epidemic methicillin-gentamicin-resistant Staphylococcus aureus in a neonatal intensive care unit.
Between October 1985 and August 1986, 49 isolates of methicillin-resistant Staphylococcus aureus (MRSA) were obtained from 26 neonates in the neonatal intensive care unit (NICU) at the Medical University Hospital, Charleston, SC. Sites of MRSA isolation were the respiratory tract (33%); nasopharynx (12%); gastrointestinal tract (12%); eye (8%); blood (6%); and catheter tips, wounds, or umbilicus (29%). Very low birth weight was a significant risk factor for MRSA acquisition. All isolates had the same phage type (47/54/75/83A), antibiogram, and whole-cell protein profile. Agarose gel electrophoresis of all 49 isolates disclosed a plasmid level of approximately 45 X 106 daltons (45 megadaltons) in ten different isolates and no plasmid DNA in 39 isolates. Cultures of NICU personnel failed to disclose MRSA carriers and environmental cultures for MRSA were negative. Ten selected isolates showed lower minimal bactericidal concentrations for hexachlorophene than for chlorhexidine. Standard infection-control measures such as contact isolation, hand washing with chlorhexidine, and cohorting (when possible) failed to contain the epidemic. Ultimately, eradication of MRSA from the NICU was associated with the institution of hexachlorophene hand washing.
Cross Infection↗
Dissemination of trimethoprim-resistant clones of Shigella sonnei in Bulgaria.
Bacterial dysentery due to Shigella sonnei remains a serious public health problem in developed countries, including Bulgaria. At the National Shigella Reference Laboratory in Sofia, 17,126 strains of S. sonnei from epidemics and sporadic cases collected from 1973 to 1987 were studied. Antibiotic susceptibility testing, phage typing, colicin typing, and biotyping were performed for all strains to allow intraspecies differentiation and to track any clonal distribution. Of all strains, 84.3% were resistant to one or more antimicrobials, the most frequent being tetracycline (Tet), streptomycin (Str), sulfonamide (Sul), chloramphenicol (Chl), ampicillin (Amp), and trimethoprim (Tmp). Resistance patterns most prevalent for successive 5-y periods included Tet, StrSulTet, and AmpChlStrSulTet, respectively. For the final 5 y, a new pattern (AmpKanStrSulTetTm [Kan = kanamycin]) was spread throughout the country by two trimethoprim-resistant clones. High-level resistance to trimethoprim (MIC greater than 1500 micrograms/mL) in both clones was determined by dihydrofolate reductase type I. The genes for trimethoprim resistance were located on a conjugative R-plasmid of approximately 145 kilobases which cotransferred all other antimicrobial resistances. A similar-sized R-plasmid had been found in earlier isolates of Bulgarian S. sonnei, suggesting that new antimicrobial resistance genes had been sequentially added to an ancestral R-plasmid. Controlling the expression of these new as well as older antimicrobial resistances, particularly for enteric pathogens, must involve reduction in usage of generic antimicrobial agents.
Ampicillin Resistance↗