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Biomedical subjects

R R Facklam

Publications and source records attributed to R R Facklam.

At least 55 records · Page 3Linked to original sources

Aerococcus urinae: intraspecies genetic and phenotypic relatedness.

A number of Aerococcus-like organisms were recently recognized as human pathogens. Five Aerococcus-like strains were proposed as members of the new species Aerococcus urinae (with type strain E2 [= NCTC 12142]) on the basis of the results of a 16S rRNA sequence analysis. The intraspecies phenotypic and genetic relatedness of 22 selected A. urinae strains was investigated, and a hitherto unrecognized esculin hydrolysis-positive biotype was identified. A total of 14 of the 15 more common esculin-negative strains exhibited very high DNA relatedness as determined by the hydroxyapatite method (the levels of relatedness were greater than 90% in 55 and 70 degrees C reactions, with 1.5% or less divergence in related sequences). The DNA relatedness among the six esculin-positive strains was more heterogeneous, and two DNA hybridization subgroups were formed. Our results are compatible with the hypothesis that both biotypes are members of the single species A. urinae, which contains two or more genetic subspecies. The putative subspecies have not been formally proposed since they cannot be definitively differentiated. The inclusion of A. urinae in the genus Aerococcus is supported by the results of 16S rRNA sequencing. The rRNA sequence data also is compatible with placing both biotypes in a single species.

Bacteriological Techniques↗

Phenotypic and genotypic characterization of Vagococcus fluvialis, including strains isolated from human sources.

This study presents phenotypic and genotypic data for seven isolates of Vagococcus fluvialis, including four strains recovered from human clinical sources, one strain isolated from an environmental source, and two strains isolated from pigs. On the basis of phenotypic characteristics, most isolates were initially classified as "unidentified enterococci," because they resembled atypical arginine-negative enterococcal species. All seven strains as well as the type strain of V. fluvialis reacted with the AccuProbe Enterococcus genetic probe. The seven isolates had virtually indistinguishable whole-cell protein profiles that were similar to that of the V. fluvialis type strain and distinct from those of Enterococcus and Lactococcus species. DNA-DNA reassociation experiments confirmed that the strains were V. fluvialis. They were 71% or more related to the V. fluvialis type strain under optimum and stringent conditions, with 2.5% or less divergence within related sequences. All strains were susceptible to ampicillin, cefotaxime, trimethoprim-sulfamethoxazole, and vancomycin and were resistant to clindamycin, lomefloxacin, and ofloxacin. Strain-to-strain variation was observed in relation to susceptibilities to 18 other antimicrobial agents. Chromosomal DNA was analyzed by pulsed-field gel electrophoresis (PFGE) after digestion with SmaI. Distinctive PFGE patterns were generated, suggesting the nonclonal nature of V. fluvialis strains. Although the number of strains was small, this report provides molecular characterization of V. fluvialis and the first evidence of a possible connection of this species with human infections.

Animals↗

Bacteremia with Streptococcus pneumoniae. Implications for therapy and prevention. Franklin County Pneumonia Study Group.

OBJECTIVE: To determine the incidence and mortality rates of patients with bacteremic infections with Streptococcus pneumoniae, and to determine the serotypes and antimicrobial susceptibilities of the pneumococcal isolates. DESIGN: Prospective case ascertainment and procurement of S pneumoniae isolates between January 1991 and April 1994. SETTING: Ten adult care hospitals in Franklin County, Ohio. PATIENTS: Patients (N = 590) in whom S pneumoniae was isolated from blood cultures. MEASUREMENTS: Demographic data from patients with pneumococcal bacteremia were obtained by chart review. Pneumococcal serotyping and antimicrobial susceptibility testing were performed on 499 bacteremic isolates. RESULTS: Among residents of Franklin County, the annual incidence of pneumococcal bacteremia was higher in patients at least 65 years old (83.0 per 100,000 population) compared with younger adults (9.6 per 100,000 population) (odds ratio [OR], 8.74; 95% confidence interval [CI], 7.19 to 10.62) and more common among African Americans than whites (OR, 1.36; 95% CI, 1.06 to 1.75). The relative risk of pneumococcal bacteremia among persons infected with the human immunodeficiency virus was 41.8 times (CI, 19.0 to 92.0) that of county residents 18 to 64 years of age. The overall mortality rate was 19% and was age-dependent, reaching 38% in patients at least 85 years old. The distribution of pneumococcal serotypes causing bacteremia was remarkably consistent over time. The incidence of drug-resistant strains increased during the study period; by 1994 14% were resistant to penicillin, 12% to ceftazidime, and 24% to trimethoprim-sulfamethoxazole. The resistant strains included several serotypes of S pneumoniae. Most serotypes (89.8%) of S pneumoniae causing bacteremia are contained in the pneumococcal vaccine. CONCLUSIONS: Increased use of pneumococcal vaccine and recognition of antimicrobial resistance patterns may assist physicians in treating patients with S pneumoniae bacteremia. Educational programs to discourage unnecessary antimicrobial drug use should be developed for patients and physicians.

Adult↗

Risk factors for carriage of drug-resistant Streptococcus pneumoniae among children in Memphis, Tennessee.

OBJECTIVES: To determine risk factors for carriage of drug-resistant Streptococcus pneumoniae to understand better the factors promoting spread of these isolates. STUDY DESIGN: We obtained medical and demographic information and nasopharyngeal swab specimens from 216 children less than 6 years old with upper respiratory tract infections, seeking medical care at five Memphis, Tenn, study sites. We evaluated risk factors for carriage of penicillin-nonsusceptible S. pneumoniae (NSSP) among 100 children with S. pneumoniae isolates. Patterns of antimicrobial prescription were recorded for enrolled children. RESULTS: Independent risk factors for carriage of NSSP included an increased number of antimicrobial treatment courses during the previous 3 months and white race. Day care attendance approached statistical significance (p = 0.07). Most children with upper respiratory tract infection received a prescription for antimicrobial drugs. These prescriptions were more common for white children than for black children. CONCLUSIONS: Increased use of antimicrobial drugs enhances the risk of carriage of NSSP. This may contribute to the higher risk among white children of NSSP infection; however, after control for antimicrobial use, white children were still at an increased risk of infection with NSSP, possibly through greater exposure to resistant strains.

Anti-Bacterial Agents↗

Invasive group B streptococcal disease: the emergence of serotype V.

Group B streptococci (GBS) cause invasive disease in neonates, pregnant adults, and nonpregnant adults with underlying or chronic disease. Previous studies found capsular serotypes Ia, Ib, II, and III cause invasive disease. Prospective population-based surveillance of invasive GBS disease was done from June 1992 to June 1993 in metropolitan Atlanta: 279 patients had invasive disease. Of these, 43% were < or = 6 months old, and 57% were adults. The incidence among all adults was 7.7/100,000/year, 33% higher than in 1989-1990 (P < .01). The incidence in nonpregnant adults was 5.9/100,000/year, 37% higher than in 1989-1990 (P < .02). Serotyping of 178 patient isolates revealed that 34% had GBS serotype Ia or Ia/c, 8% had Ib/c, 6% had II or II/c, 29% had III, 0% had IV, 21% had V, and 2% were nontypeable. Serotype V was recovered from all groups and was the most common serotype from nonpregnant adults. Serotype V isolates appeared to be highly related genetically. The increasing incidence of GBS disease in adults, the changing distribution of serotypes, and the emergence of serotype V will impact vaccine strategies.

Adolescent↗

Spread of multidrug-resistant Streptococcus pneumoniae among hospitalized children in Slovakia.

A multidrug-resistant serotype 14 strain of Streptococcus pneumoniae was isolated from sterile-site specimens and nasopharyngeal secretions from > 200 children in Slovakia between 1985 and 1990. Nasopharyngeal culture surveys were done to determine the extent of spread and means of transmission of this strain. The resistant strain was isolated from cultures of 8 (33.0%) of 24 children at hospital A and from 1 (0.8%) of 130 children attending outpatient clinics or day care centers (P < .001). One-quarter of the initially uncolonized children at hospital A acquired the resistant strain during hospitalization. Among hospitalized children, frequent antimicrobial drug use (P < .01), prior hospitalization (P < .005), and length of hospital stay (P < .001) were associated with infection with the resistant strain. These findings support limiting broad-spectrum antimicrobial drug use and nonessential hospitalizations in settings were drug-resistant pneumococci are prevalent. Development of a pneumococcal vaccine that is immunogenic in young children is urgently needed.

Adolescent↗

The continued emergence of drug-resistant Streptococcus pneumoniae in the United States: an update from the Centers for Disease Control and Prevention's Pneumococcal Sentinel Surveillance System.

As part of ongoing national surveillance, serotyping and antimicrobial susceptibility testing were done on all pneumococcal isolates recovered from normally sterile body sites of patients at 12 hospitals in 11 states during 1993-1994. Of 740 isolates, 14.1% were penicillin-nonsusceptible Streptococcus pneumoniae (PNSP; MIC > or = 0.1 microgram/mL), 3.2% were penicillin-resistant (MIC > or = 2.0 micrograms/mL), and 25.5% were nonsusceptible to more than one antimicrobial agent. PNSP were more prevalent among children < 6 years old (18.4%) than patients > or = 18 years old (11.7%) and among white persons (16.2%) than black persons (12.1%). PNSP represented 15 serotypes, but 89% of PNSP were serotypes in the 23-valent pneumococcal vaccine. The proportion of isolates with reduced susceptibility and the number of serotypes of nonsusceptible strains are increasing in the United States. Improved local surveillance for PNSP infections, judicious use of antibiotics, and development and use of effective pneumococcal vaccines will be required to treat and prevent disease caused by these strains.

Adolescent↗

Risk factors for nasopharyngeal carriage of resistant Streptococcus pneumoniae and detection of a multiply resistant clone among children living in the Yukon-Kuskokwim Delta region of Alaska.

BACKGROUND: Children < 2 years old living in the Yukon-Kuskokwim Delta (YKD) region of Alaska have one of the highest pneumococcal bacteremia rates of in the world. METHODS: To determine the prevalence of and risk factors for infection with intermediate or resistant Streptococcus pneumoniae in the YKD, we cultured nasopharyngeal secretions of healthy children < or = 5 years old, reviewed their hospital records and administered questionnaires to accompanying parents. RESULTS: Of 185 children evaluated we obtained 95 pneumococcal isolates; drug susceptibility patterns and serotyping results were available for 92. Of these, 33 (36%) were intermediate or resistant to at least one drug class tested; 27 isolates were intermediate (minimum inhibitory concentration 0.1 to 1.0 mg/l) and none were resistant to penicillin. Compared with other isolates, capsular serotype 6B isolates were more likely to be intermediate or resistant to at least one drug (relative risk, 5.3; P < 0.001) and to more than one drug (relative risk, 17.0; P < 0.001). The majority of 6B isolates had identical pneumococcal surface protein A patterns. Carriage of intermediate or resistant pneumococcus was associated with age < 2 years (relative risk, 3.0; P < 0.001) but not with antibiotic use or other evaluated risk factors. CONCLUSIONS: Young age but not antibiotic use was associated with carriage of intermediate or resistant S. pneumoniae in the YKD region of Alaska. Much of the intermediate or resistant pneumococcus in the YKD may have resulted from the proliferation of a single capsular serotype 6B clone.

Alaska↗

Phenotypic and genotypic characterization of atypical Lactococcus garvieae strains isolated from water buffalos with subclinical mastitis and confirmation of L. garvieae as a senior subjective synonym of Enterococcus seriolicida.

During a survey of bacterial agents that cause subclinical mastitis in water buffalos, we isolated several strains of gram-positive cocci that appeared to be enterococci except that they grew very slowly at 45 degrees C and grew slowly in broth containing 6.5% NaCl. On the basis of the results of conventional physiologic tests, these strains were identified as Enterococcus durans. However, none of the strains reacted with the AccuProbe Enterococcus genetic probe. The whole-cell protein profiles of these organisms were compared with the profiles of Enterococcus and Lactococcus reference strains. apart from minor quantitative differences, the mastitis isolates had indistinguishable protein profiles that were similar to the profiles of the Lactococcus garvieae and Enterococcus seriolicida type strains. The results of DNA relatedness studies performed by using the hydroxyapatite method at 55 and 70 degrees C indicated that all of the mastitis isolates were related to the type strain of L. garvieae at the species level, despite the fact that they exhibited several uncommon phenotypic characteristics (growth at 45 degrees C, growth in broth containing 6.5% NaCl, and failure to produce acid from mannitol and sucrose). The high levels of DNA relatedness between strains of L. garvieae is a senior synonym of E. seriolicida, L. garvieae should be retained as the species name and strain ATCC 43921 should remain the type strain of this species.

Animals↗

Atmospheric growth requirements for Alloiococcus species and related gram-positive cocci.

The growth of Alloiococcus otitis under different atmospheres and nutritional conditions was studied. The growth rates of 25 strains of gram-positive cocci representing five genera on heart infusion agar plates containing 5% rabbit blood and on brucella agar plates with and without sheep blood under aerobic, increased CO2, and anaerobic atmospheres were compared. Eight strains of alloiococci plated on heart infusion agar with rabbit blood and on brucella sheep blood agar grew under aerobic and candle jar atmospheres. Two of these strains showed poor anaerobic growth after 7 days. Strains of Aerococcus viridans, Aerococcus urinae, Helcococci kunzi, Dolosigranulum pigrum, Gemella haemolysans, and Gemella morbillorum grew well under all three atmospheres and on the three types of media and in thioglycolate broth. These results confirm all the aerobic atmospheric requirements for Alloiococcus strains and show that aerobic growth characteristics help distinguish the alloiococci from the other gram-positive cocci that are facultatively anaerobic.

Aerobiosis↗

Antimicrobial susceptibilities of Lactococcus lactis and Lactococcus garvieae and a proposed method to discriminate between them.

The MICs of antimicrobial agents contained in the SCEPTOR Streptococcus MIC panels (Becton Dickinson Microbiology Systems) were determined for Lactococcus lactis, L. garvieae, and unknown Lactococcus species. Several isolates had reduced susceptibilities to many of the antimicrobial agents contained in the panel. For L. garvieae, the MICs of penicillin and, possibly, cephalothin were higher than for L. lactis, and unlike L. lactis, L. garvieae was resistant to clindamycin, indicating that knowledge of the Lactococcus species causing an infection might influence the choice of antimicrobial therapy. Susceptibility to clindamycin can also be used to differentiate between L. lactis and L. garvieae.

Bacteriological Techniques↗

The prevalence of drug-resistant Streptococcus pneumoniae in Atlanta.

BACKGROUND: Streptococcus pneumoniae is a major cause of illness, and the emergence of drug-resistant strains threatens to complicate the management of pneumococcal infections. We conducted a laboratory-based surveillance for drug-resistant S. pneumoniae among patients with invasive pneumococcal infections in Atlanta. METHODS: From January through October 1994, pneumococcal isolates from 431 patients with invasive disease in metropolitan Atlanta were serotyped and tested to determine their susceptibility to various antimicrobial agents. Susceptibility to the antimicrobial agents was defined according to guidelines established by the National Committee for Clinical Laboratory Standards. RESULTS: The annual incidence of invasive pneumococcal infection was 30 cases per 100,000 population. Isolates from 25 percent of the patients were resistant to penicillin (7 percent were highly resistant), and isolates from 26 percent were resistant to trimethoprim-sulfamethoxazole (7 percent highly resistant). Fifteen percent of the isolates were resistant to erythromycin, 9 percent to cefotaxime (4 percent were highly resistant), and 25 percent to multiple drugs. Drug-resistant pneumococci were found in both children and adults. Children under six years of age were more likely than older children and adults to have isolates resistant to multiple drugs or cefotaxime. Whites were more likely than blacks to have invasive pneumococcal infections caused by drug-resistant organisms. Among white children younger than six years, 41 percent of the S. pneumoniae isolates were resistant to penicillin. CONCLUSIONS: Drug-resistant strains of S. pneumoniae are common among both children and adults in Atlanta. Although blacks had a higher incidence of invasive pneumococcal infections than whites, whites were more likely to be infected with a drug-resistant isolate. Control of drug-resistant pneumococci will require more judicious use of antimicrobial agents and wider use of the pneumococcal polysaccharide vaccine.

Adolescent↗

Discrimination of Streptococcus pneumoniae from other upper respiratory tract streptococci by arbitrarily primed PCR.

OBJECTIVES: In the clinical laboratory, identification of Streptococcus pneumoniae can be confused with other streptococci. Conventional biochemical tests such as optochin sensitivity and bile solubility can give inconsistent results. This report presents a method to distinguish true S. pneumoniae from other upper respiratory tract streptococci when conventional tests fail. DESIGN AND METHODS: We used arbitrarily primed polymerase chain reaction with the single primer M13 universal as a method to distinguish S. pneumoniae from other upper respiratory tract streptococci. RESULTS: The fingerprint pattern of S. pneumoniae was established by amplifying DNA of S. pneumoniae type strains 1-48 and of other common upper respiratory tract streptococci at three different DNA concentrations with the single primer M13 universal. From these type strains, a common arbitrarily primed-polymerase chain reaction pattern was identified characterized by two predominant bands of equal intensity at 800 base pairs and at 1100 base pairs. Fingerprint patterns of viridans streptococci were easily distinguishable from those of S. pneumoniae. Many of the clinical isolates used in this study were equivocal by conventional tests but were distinguishable by their fingerprint patterns. CONCLUSIONS: Our results indicated that the fingerprint pattern of S. pneumoniae is species specific and distinguishes true S. pneumoniae of clinical isolates from other streptococci when conventional biochemical tests are unclear.

Bacterial Typing Techniques↗

Serotype distribution of Streptococcus pneumoniae infections among preschool children in the United States, 1978-1994: implications for development of a conjugate vaccine.

Conjugation of pneumococcal polysaccharide antigens to a protein carrier may improve protective immunity after vaccination of young children, an age group with high incidence of Streptococcus pneumoniae infection and poor immune responses to polysaccharide vaccines. To identify serotypes most commonly associated with infection in young children, pneumococcal isolates were serotyped from 3884 children < 6 years old (including 3007 < 2 years old) with pneumococcal bacteremia (n = 3169), meningitis (n = 401), or otitis media (n = 314). The isolates were submitted as part of a national surveillance during 1978-1994. Seven serotypes (14, 6B, 19F, 18C, 23F, 4, and 9V) accounted for 3045 isolates (78%). A conjugate pneumococcal vaccine protecting against these seven serotypes and serologically cross-reactive serotypes could potentially prevent 86% of bacteremia and 83% of meningitis but only 65% of otitis media cases. The proportion of isolates covered by such a vaccine increased from 78% to 87% during 1978-1994. Surveillance for pneumococcal serotypes causing infection is needed to detect shifts in serotype distribution over time.

Bacterial Vaccines↗

Multiple antimicrobial resistance of pneumococci in children with otitis media, bacteremia, and meningitis in Slovakia.

Penicillin-resistant pneumococci have been isolated from middle ear fluid, blood, cerebrospinal fluid, and nasopharyngeal secretions of several hundred children in Slovakia since 1985; 116 of these isolates were serotyped and tested for susceptibility to antimicrobial drugs at the Centers for Disease Control and Prevention. To define the prevalence of drug-resistant pneumococci and identify risk factors for infection, laboratory and medical records were reviewed. Nearly all (96%) of the resistant strains tested were serotype 14. Of these, all were resistant to penicillin (MIC, 4-16 micrograms/mL); most were resistant to cefaclor, erythromycin, tetracycline, and chloramphenicol; and many had decreased susceptibility to trimethoprim-sulfamethoxazole and ceftriaxone. Frequent antibiotic use, prior hospitalization, and length of hospital stay (P < .001 for all 3) were associated with infection with resistant strains. These findings suggest the need for routine screening of pneumococcal isolates for penicillin resistance and highlight the importance of controlling globally the spread of resistant pneumococci.

Bacteremia↗