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Biomedical subjects

J M Janda

Publications and source records attributed to J M Janda.

At least 19 recordsLinked to original sources

Association of Neisseria cinerea with ocular infections in paediatric patients.

Twenty-two strains of Neisseria cinerea were recovered from paediatric patients over a 7-year period and forwarded to the Microbial Diseases Laboratory for biochemical identification and/or confirmation. Eighteen of these 22 strains (82%) were recovered from the eyes of very young children (< or = 1 year), > 50% occurring during the neonatal period. The majority of eye isolates were involved in a variety of ocular infections including orbital cellulitis, conjunctivitis, and eye discharge (most common); in four of the 13 instances (31%) where laboratory data was available, Neisseria cinerea was recovered in pure culture. Neisseria cinerea isolates were often submitted to the Microbial Diseases Laboratory as possible 'N. gonorrhoeae' or 'Neisseria species' due to problems resulting from the use of commercial assays or unfamiliarity with the organism. These observations indicate that N. cinerea can produce eye infections in very young children, who presumably acquire this organism vertically from the mother during birth. Accurate identification of N. cinerea in such infants can preclude the social trauma and possible legal ramifications which can initially result from its misidentification as N. gonorrhoeae.

Eye

Biochemical and pathogenic properties of Shewanella alga and Shewanella putrefaciens.

We characterized 49 strains of Shewanella spp. from clinical (n = 31) and nonhuman (n = 18) sources. Most Shewanella alga organisms (Gilardi biovar 2; Centers for Disease Control and Prevention [CDC] biotype 2) originated from clinical material (92%), failed to produce acid from carbohydrates other than D-ribose, and were biochemically and enzymatically fairly homogeneous. In contrast, Shewanella putrefaciens organisms (Gilardi biovars 1 and 3; CDC biotype 1) were more often associated with nonhuman sources (70%), were able to utilize a number of sugars (sucrose, L-arabinose, and maltose), and were found to exhibit wider variations in biochemical characteristics; three biotypes within S. putrefaciens were detected. Notable differences between the two species in enzymatic activity, determined with the API-ZYM system (bioMérieux, Hazelwood, Mo.), and cellular fatty acid profiles, determined by the MIDI system (Microbial ID Inc., Newark, Del.), were also detected. Pathogenicity studies of mice indicate that S. alga appears to be the more virulent species, possibly due to the production of a hemolytic substance.

Animals

Overview of the etiology of wound infections with particular emphasis on community-acquired illnesses.

Wound cultures represent a general catchall category for a group of extremely diverse anatomic samples that range from superficial specimens of cutaneous structures (folliculitis, cellulitis) to specimens revealing invasive infections involving deep fascial planes and muscle (myonecrosis). Because of the complex nature of these infective processes, the terminology associated with such infections is often imprecise and confusing. Wounds are the result of trauma, either intentionally or accidentally induced. Nosocomial wound infections result primarily from surgical procedures, the development of pressure sores, or catheterization. Community-acquired wound infections are often preceded by injuries resulting from occupational exposure or recreational activities and are associated with a greater diversity of microorganisms due to the exposure of open wounds to inhabitants of the microbial biosphere. This review provides a general overview of the categories of wound infections and describes their acquisition and clinical significance. Particular emphasis is placed on selected community-acquired wound infections and the etiologic agents associated with such conditions.

Animals

Four additional cases of Burkholderia gladioli infection with microbiological correlates and review.

Burkholderia gladioli has only recently been reported to be a human pathogen. Four cases of B. gladioli infection (including bacteremia, pneumonia, and cervical adenitis) in two adults and two young children are reported. Three of these four patients were severely immunocompromised. Commercial systems were frequently unable to identify this bacterium correctly. Antimicrobial susceptibility patterns indicated that B. gladioli strains were susceptible to the quinolones, aminoglycosides, and imipenem. In vitro laboratory investigations demonstrated that B. gladioli strains were susceptible to complement-mediated lysis of pooled human serum, thus implying that healthy individuals should be immune to infection. These four cases together with three previously reported cases suggest that B. gladioli primarily causes disease in severely immunocompromised individuals. The lack of mortality associated with infection, coupled with susceptibility to serum and lack of recognizable virulence-associated factors, suggests that this species has a low pathogenic potential.

Adult

Enterobacter cancerogenus ("Enterobacter taylorae") infections associated with severe trauma or crush injuries.

Five cases of Enterobacter cancerogenus infections (wound, n = 4; bacteremia, n = 1) in adults are described. All infections seemed to be community acquired and occurred after precipitating events such as multiple trauma to the head or severe crush injuries. All five strains of E cancerogenus were recovered in pure culture, and three of these were isolated on multiple occasions. The results indicate that E cancerogenus can cause wound infections and septicemia in persons environmentally exposed to these organisms during traumatic events.

Adolescent

Multicenter evaluation of the MicroScan Rapid Gram-Negative Identification Type 3 Panel.

The accuracy and performance of the revised MicroScan Rapid Gram-Negative Identification Type 3 Panel (Dade MicroScan Inc., West Sacramento, Calif.) were examined in a multicenter evaluation. The revised panel database includes data for 119 taxa covering a total of 150 species, with data for 12 new species added. Testing was performed in three phases: the efficacy, challenge, and reproducibility testing phases. A total of 405 fresh and stock gram-negative isolates comprising 54 species were tested in the efficacy phase; 96.8% of these species were identified correctly in comparison to the identification obtained either with the API 20E system (bioMérieux Vitek, Hazelwood, Mo.) or by the conventional tube method. The number of correctly identified isolates in the challenge phase, including new species added to the database, was 221 of 247, or 89.5%, in comparison to the number correctly identified by the conventional tube method. A total of 465 isolates were examined for intra- and interlaboratory identification reproducibility and gave an agreement of 464 of 465, or 99.8%. The overall reproducibility of each individual identification test or substrate was 14,373 of 14,384, or 99.9%. The new Rapid Gram-Negative Identification Type 3 Panel gave accurate and highly reproducible results in this multiple-laboratory evaluation.

Evaluation Studies as Topic

Analysis of the thermostable direct hemolysin (tdh) gene and the tdh-related hemolysin (trh) genes in urease-positive strains of Vibrio parahaemolyticus isolated on the West Coast of the United States.

Urease-positive (Ure+) and urease-negative (Ure-) strains of Vibrio parahaemolyticus isolated from patients on the West Coast of the United States between 1979 and 1995 were analyzed for the thermostable direct hemolysin (tdh) gene and the tdh-related hemolysin (trh) genes (trh1 and trh2). The DNA colony hybridization method with the polynucleotide probes was used to determine the distribution of the genes. Of 60 Ure+ strains, 59 strains (98%) had the trh (either trh1 or trh2) gene and 54 strains (90%) carried the tdh gene. The absence of the trh gene or a related sequence in an exceptional Ure+ strain was confirmed by Southern blot analyses. The stronger correlation with the trh gene than with the tdh gene was mostly attributable to strains possessing only the trh2 gene. Of 25 Ure- strains, 20 strains (80%) had the tdh gene but none had the trh gene. These results indicate a very strong correlation between the Ure+ phenotype and the trh gene and are consistent with those reported for strains isolated in Asia. The Ure+ strains carrying the trh genes were not restricted to a unique group of the strains. The O4:K12 strains carrying the trh1 gene have predominantly been isolated since 1979. However, strains of various non-O4:K12 serovars carrying either the trh1 or the trh2 gene became predominant after 1992. In addition, analysis by the arbitrarily primed PCR method revealed two subgroups within the selected Ure+ O4:K12 strains. Hybridization tests with oligonucleotide probes demonstrated that the trh1 sequences of the West Coast strains differ to some extent from those of Asian strains. Nevertheless, a PCR method previously established to detect both the trh1 and the trh2 genes in Asian strains could detect 98% of those genes in the West Coast strains.

Bacterial Proteins

Iron utilization studies in Citrobacter species.

Seventy-one strains of Citrobacter were screened for iron scavenging mechanisms by biologic and chemical assays. Essentially all citrobacteria (70/71) were found to elaborate enterobactin-like siderophores by both biologic and chemical assays, however only c. koseri (C. diversus) was found to produce aerobactin. The concentration of ethylenediamine di(o-hydroxyphenylacetic acid) (EDDA) required to inhibit the growth of individual Citrobacter strains by depleting free iron ranged from 250 micrograms/ml to 100 micrograms/ml. Iron utilization studies of selected citrobacter isolates indicated that hemin and hematin could reverse the effects of iron limitation on growth under iron-stressed conditions (1000 micrograms/ml of EDDA). Two C. koseri strains grown under iron-restricted conditions showed similar changes in their whole cell protein profiles including induction of high molecular mass proteins (72-83 kDa) which may play a role in iron acquisition under iron-stressed conditions. The collective results support an additional virulence-associated mechanism for C. koseri strains which may help explain the greater pathogenic potential this group has for causing serious extraintestinal disease in humans.

Bacterial Proteins

Characterization of Aeromonas hydrophila strains of clinical, animal, and environmental origin expressing the O:34 antigen.

A collection of Aeromonas strains of different origins were characterized for isolates expressing the O:34 somatic antigen. Of over 200 strains tested, approximately 14% belonged to serogroup O:34 with >85% of these strains identified as A. hydrophila regardless of source. A subset of 14 A. hydrophila O:34 strains were further analyzed for a number of structural and pathogenic features. Most O:34 strains expressed similar whole-cell protein profiles with regards to minor bands, but major band differences were noted in outer membrane proteins (OMPs) migrating between the 31K and 58K region. OMP profiles could be subdivided into three distinct patterns. All O:34 strains expressed a heterogeneous O polysaccharide side chain profile in their lipopolysaccharide (LPS), although some variation in the electrophoretic migration of lower and higher molecular weight LPS bands was noted. Polyclonal antisera raised against a 45-K OMP-associated protein of one O:34 strain (AH-195) reacted in immunoblot assays with a major 43 to 46-K OMP in 11 of 14 (79%) O:34 strains tested. Most O:34 strains (69%) were found to be pathogenic in mice with LD-50 values (i.p.) of <1.0 x 10(7) CFU; pathogenicity appeared to correlate best with elevated protease activity. The collective results suggest significant differences in both structural and pathogenic properties between some members of the O:34 group originating from human and nonhuman (fish, water) sources.

Aeromonas hydrophila

An initial appraisal of the clinical significance of Roseomonas species associated with human infections.

We reviewed laboratory, clinical, and epidemiologic data on 35 patients from whom organisms belonging to the genus Roseomonas, a pink-pigmented gram-negative coccobacillus, were isolated over a 22-year period (1972-1994). Roseomonas strains were most commonly isolated from middle-aged women with one of several underlying conditions, including cancer and diabetes. Roseomonas was most commonly isolated from the blood, in association with clinical signs of sepsis. Approximately 60% of all isolates were judged to be of possible clinical significance, either as primary or secondary pathogens; 75% of all strains were recovered in pure culture. Roseomonas gilardii was the most frequently isolated species and was significantly associated with septicemia and underlying immunocompromised conditions; the species of 29% of all Roseomonas isolates could not be unequivocally identified with presently available differential tests. Genomospecies 5, currently an unnamed taxon within the genus Roseomonas, was primarily recovered as a commensal from young adults attending a sexually transmitted diseases clinic. The findings suggest that although this genus appears to have an overall low pathogenic potential for humans, Roseomonas species-in particular, R. gilardii-may be significant pathogens in persons with underlying medical complications.

Adolescent

The thermostable direct hemolysin gene (tdh) of Vibrio hollisae is dissimilar in prevalence to and phylogenetically distant from the tdh genes of other vibrios: implications in the horizontal transfer of the tdh gene.

Vibrio hollisae strains isolated recently from patients in various locations were examined for the presence of the thermostable direct hemolysin gene (tdh) using nucleic acid hybridization and polymerase chain reaction assays. The results were consistent with the previous finding that all strains of V. hollisae carry the tdh gene. In contrast, the tdh gene has been detected in a minority of strains for other Vibrio species (V. parahaemolyticus, V. cholerae non-O1, and V. mimicus). Detailed phylogenetic analysis showed that the tdh genes of the non-V. hollisae species were very closely related to each other and that the tdh gene of V. hollisae was distantly related to the tdh genes of the non-V. hollisae species. These results and the proposed insertion sequence-mediated tdh transfer mechanism suggest that the tdh gene may have been maintained stably in V. hollisae and that the tdh genes of the non-V. hollisae species may have been involved in recent horizontal transfer.

Bacterial Proteins

Biochemical investigations of biogroups and subspecies of Morganella morganii.

We determined the subspecies and biogroup designations for 73 strains of Morganella morganii principally recovered from routine clinical specimens. On the basis of trehalose fermentation, 90% of all strains were identified as M. morganii subsp. morganii (trehalose negative), while the remaining 10% were designated M. morganii subsp. sibonii (trehalose positive). Using three tests (ornithine decarboxylase [ODC] and lysine decarboxylase [LDC] activities and susceptibility to tetracycline), we determined the biogroup designations for these 73 strains. Four of the seven recognized biogroups within the genus Morganella were found in the study, with biogroup A (ODC positive [ODC+], LDC negative [LDC-]) predominating (78%); all M. morganii subsp. sibonii strains were found to belong to biogroup G (ODC+, LDC-). Rapid glycerol fermentation (24 h) was linked to nonmotility and biogroup B strains (ODC+, LDC+). LDC activity but not tetracycline resistance appeared to be associated with the possession of a 40- to 45-MDa plasmid. The use of three commercial systems (API ZYM, API 50 CH, and Biolog GN) failed to detect any new biochemical tests useful for subspecies identification, with the possible exception of L-phenylalanine utilization as a sole carbon source in the Biolog GN system. No Morganella strain was found to invade either HEp-2 or Vero cell lines, but four of seven M. morganii subsp. morganii strains were cytotoxic on sheets of both cells. This cytotoxic activity appeared to correlate with the rapid expression of beta-hemolytic activity.

Animals

Further studies on biochemical characteristics and serologic properties of the genus Aeromonas.

We characterized a collection of 268 Aeromonas isolates from diverse sources (clinical, animal, and environmental sources) for their species and serogroup designations. Overall, 97% of these strains could be identified to the genomospecies level by using an expanded battery of biochemical tests. Members of the Aeromonas hydrophila complex (A. hydrophila, HG2, and A. salmonicida), a group that has previously been difficult to separate biochemically, could easily be distinguished from one another by using a number of recently described phenotypic properties which included utilization of DL-lactate and urocanic acid. Differences in species distributions on the basis of the source of isolation were noted. Serogroup analysis of these 268 isolates plus a number of reference cultures indicated that (i) each genomospecies is serologically heterogeneous and individual serogroups can be found in more than one species, (ii) most type or reference strains for each hybridization group are not serologically representative of the genomospecies at large, (iii) serogroups O:11, O:34, and O:16 predominate clinically (48%), supporting previous studies indicating their importance in human infections, and (iv) most A. trota strains do not express the O139 antigen of Vibrio cholerae. The collective results suggest that both species and serogroup designations are important factors in establishing which isolates can cause human infections when they are acquired from nonclinical sources (foods, animals, and the environment).

Aeromonas

Difficulties encountered in identification of a nutritionally deficient streptococcus on the basis of its failure to revert to streptococcal morphology.

Difficulties were encountered in the recognition of a nutritionally deficient streptococcus which continued to display aberrant morphologic forms (especially bulbous swellings and filament formation) despite provision of growth factors. With isolates displaying diverse morphologic entities not characteristic of a given species, e.g., Streptobacillus moniliformis or Erysipelothrix rhusiopathiae, nutritionally deficient streptococci should be considered.

Bacteremia

Photochemical inactivation of pathogenic bacteria in human platelet concentrates.

Platelet concentrates (PC) may be infrequently contaminated with low levels of bacteria that can cause septicemia and death in patients receiving transfusion therapy. We evaluated the efficacy of a photochemical decontamination (PCD) technique using 8-methoxypsoralen (8-MOP) and long wavelength UV light (UVA) to inactivate bacteria in standard therapeutic PC. Twelve phylogenetically distinct pathogenic bacteria, 5 gram-positive and 7 gram-negative organisms, were seeded into PC to a final challenge dose ranging from 10(5) to 10(7) colony-forming units (CFU)/mL. Contaminated PC were treated with 8-MOP (5 micrograms/mL) and 5 J/cm2 of UVA, a PCD treatment regimen found to adequately preserve in vitro platelet function. Greater than 10(5) CFU/mL of all 5 gram-positive (Staphylococcus aureus, Streptococcus epidermidis, Streptococcus pyogenes, Listeria monocytogenes, and Corynebacterium minutissimum) and 2 of the gram-negative (Escherichia coli and Yersinia enterocolitica) organisms were inactivated. The remaining 5 gram-negative organisms were more resistant, with less than 10(1) to 10(3.7) CFU/mL inactivated under these conditions. The inactivation efficiency for this resistant group of gram-negative organisms was improved when PC were resuspended in a synthetic storage medium with reduced plasma protein concentration (15%) and an increased 8-MOP concentration (23.4 micrograms/mL). Illumination with 3 J/cm2 of UVA in this system inactivated greater than 10(5) CFU/mL of 4 resistant gram-negative organisms (Salmonella choleraesuis, Enterobacter cloacae, Serratia marcescens, and Klebsiella pneumoniae) and 10(4.1) CFU/mL of the most resistant gram-negative organism (Pseudomonas aeruginosa). This level of PCD treatment did not adversely affect in vitro platelet function. These results demonstrate that PCD using 8-MOP (5 to 23.4 micrograms/mL) effectively inactivated high levels of pathogenic bacteria in PC with adequate preservation of in vitro platelet properties.

Adult

Severe gastroenteritis associated with Vibrio hollisae infection: report of two cases and review.

Vibrio hollisae, one of the more recently described halophilic Vibrio species, is infrequently associated with gastrointestinal disease and only rarely recovered from individuals presenting with gram-negative sepsis. In this report we describe two cases of severe gastrointestinal disease associated with V. hollisae in otherwise healthy individuals. In one of these individuals, severe epigastric pain was apparently associated with signs of pseudoappendicitis, necessitating exploratory surgery. In both individuals, infection was associated with the ingestion of raw shellfish. These cases are discussed in light of previous reports on the epidemiology, pathogenesis, and spectrum of disease caused by this unusual pathogen.

Adult

Aeromonas species in septicemia: laboratory characteristics and clinical observations.

We retrospectively analyzed clinical and epidemiological data on and laboratory characteristics of 53 cases of aeromonas septicemia. Only four Aeromonas genomospecies (species defined by DNA relatedness) were associated with the 53 cases, with Aeromonas hydrophila (sensu stricto) predominating (47%). Nearly 60% of all Aeromonas isolates from blood fell into one of four somatic groups: serogroups O:11, O:16, O:18, and O:34. Unlike Aeromonas-associated gastroenteritis, septicemia did not peak in frequency during the warmer months but rather was most common in January through March, when approximately 40% of cases occurred. In vitro tests of the pathogenicity of 20 selected blood isolates of Aeromonas indicated that resistance to complement-mediated lysis, elevated levels of protease and hemolysin activity, and the ability to elaborate siderophores correlated with higher virulence. Species and serogroup designations also correlated with the degree of virulence. Susceptibility studies of 50 strains indicated that A. hydrophila was the most drug-resistant species and that Aeromonas veronii was the most susceptible. Susceptibility to first- and second-generation cephalosporins and carbenicillin was species-associated.

Adolescent