Jill Clarridge

Clostridium difficile (CD) disease remains a costly and important hospital associated infection. Although nontoxigenic CD is detected by some CD testing methods, can interfere with some detection algorithms and has been suggested as a treatment for CD disease, little is known about the relative occurrence of toxigenic and nontoxigenic CD in a single institution. We used both chromogenic and selective agar medium to recover Clostridium difficile (CD) isolates and a molecular method to detect the toxin B gene from over 2400 fresh unformed stool specimens with isolates further tested for the toxin B gene. We recovered 74 nontoxigenic and 306 toxigenic CD isolates for which a collection site could be assigned. The frequency of recovery of toxigenic and nontoxigenic CD for each hospital location and the ratio of toxigenic to nontoxigenic CD were calculated. Although the overall prevalence of toxigenic and nontoxigenic CD was 12.7% and 3.1%, respectively, on some wards 48% of all CD were ...

We determined the frequency distribution of Actinomyces spp. recovered in a routine clinical laboratory and investigated the clinical significance of accurate identification to the species level. We identified 92 clinical strains of Actinomyces, including 13 strains in the related Arcanobacterium-Actinobaculum taxon, by 16S rRNA gene sequence analysis and recorded their biotypes, sources, and disease associations. The clinical isolates clustered into

Mupirocin is a topical antimicrobial used to decolonize patients who carry methicillin resistant Staphylococcus aureus (MRSA), and the topical agent retapamulin may be a potential alternative therapy. The goal of this study was to determine the in vitro activity of retapamulin in a naïve population along with a panel of fifteen antimicrobial agents, including mupirocin, for 403 MRSA isolates collected longitudinally at the Veterans Affairs Puget Sound Health Care System. The minimum inhibitory concentration (MIC) for retapamulin had a unimodal distribution ranging from 0.008-0.5 μg/ml. One isolate had an MIC of >16 μg/ml, was also resistant to clindamycin and erythromycin, and was recovered from the nares of a patient receiving hemodialysis. 24 (6%) and 11 (3%) isolates, respectively, demonstrated low level resistance (8-64 μg/ml) and high level resistance (≥512 μg/ml) to mupirocin. Isolates were recovered from 10 patients both pre- and post- mupirocin therapy. Of those, 2 patien...

We have isolated two phenotypically distinct nonfastidious Francisella strains (Fx1 and Fx2) from the blood of compromised patients with pneumonia and compared them with eight other Francisella strains, including Francisella tularensis biovar tularensis, F. tularensis biovar novicida, and F. philomiragia. Our isolates grew well on sheep blood agar, chocolate agar, modified Thayer-Martin agar, and Trypticase soy agar. Fx1 and Fx2 were determined to be within the Francisella genus by cellular fatty acid analysis and by the utilization of glucose, production of H2S and catalase, and lack of motility, oxidase, nitrate reductase, and gelatinase. They were additionally shown to belong to the species F. tularensis by sequencing of two variable regions comprising approximately 500 nucleotides of the 16S rRNA gene. Also, RNA probe hybridization confirmed their belonging to the species F. tularensis. However, the new strains, which are not identical, are distinguished from other F. tularensis...

Previous studies have demonstrated that the four subspecies of the human pathogen Francisella tularensis, despite showing marked variations in their virulence for mammals and originating from different regions in the Northern Hemisphere, display a very close phylogenetic relationship. This property has hampered the development of generally applicable typing methods. To overcome this problem, we evaluated the use of PCR for discrimination of the subspecies using various forms of long arbitrary primers or primers specific for repetitive extragenic palindromic sequences (REP) or enterobacterial repetitive intragenic consensus (ERIC) sequences. Patterns generated by use of REP, ERIC, or long arbitrary primers allowed differentiation at the species level and of the four subspecies of F. tularensis. With each of these three methods, similar or identical clustering of strains was found, and groups of strains of different geographical origins or differing in virulence showed distinct patter...

The recent observation that Branhamella catarrhalis may cause a variety of infections in humans has stimulated interest in human host defenses against this organism. We encountered 21 patients with B. catarrhalis infection: seven with pneumonia, 13 with a purulent exacerbation of chronic bronchitis, and one with purulent sinusitis. Normal human serum (NHS) demonstrated no bactericidal activity against 20 of the 21 isolates. In contrast, 7 of 19 acute and 18 of 20 convalescent sera demonstrated significant bactericidal effects against the corresponding B. catarrhalis isolate. Heating convalescent sera to 56 C for 30 min abolished bactericidal activity. This activity was restored by NHS but not by complement-rich guinea pig serum. Selective blockage of the classic complement pathway eliminated bactericidal activity, whereas selective blockage of the alternative pathway did not. IgG isolated from convalescent serum plus NHS was bactericidal for the corresponding B. catarrhalis isolate. These results suggest that most patients with pulmonary infections due to B. catarrhalis develop a convalescent IgG antibody response that mediates serum bactericidal activity by the classic complement pathway.

Bordetella avium is thought to be strictly an avian pathogen. However, 16S rRNA gene sequencing identified 2 isolates from 2 humans with respiratory disease as B. avium and a novel B. avium-like strain. Thus, B. avium and B. avium-like organisms are rare opportunistic human pathogens.

Streptococcus dysgalactiae subspecies equisimilis (SDSE) are isolated from the throat of patients with pharyngitis, although the clinical significance remains debated. We sought to determine the incidence and association with pharyngitis of SDSE in an adult veteran population. Organisms were phenotypically identified to subspecies and Lancefield group, with selective 16S rRNA gene sequencing. From 833 throat cultures, the overall frequency of SDSE was 3.4% (64% group C and 36% group G) as compared to 8.6% for S. pyogenes (GAS). SDSE was described as a large colony in only 29% of the original culture evaluations by bench technologists, and clinical symptoms were similar for GAS and SDSE. Laboratory algorithms that are limited to identification of only GAS or are based on Lancefield group or visual identification of "large-colony type" β hemolytic Lancefield group C and G streptococci may be missing or misidentifying SDSE along with Anginosus group streptococci.

A rapidly progressing panophthalmitis due to Bacillus cereus developed in three patients. Infection was associated with intravenous drug abuse in two patients and was traced to contaminated injection paraphernalia in one. In the third patient, infection was associated with a foreign-body injury to the eye. Anterior chamber aspiration revealed the organism on Gram's stain in one case and isolation of the bacteria in all three. Despite intravenous and intraocular antibiotic therapy, the infection progressed rapidly and resulted in enucleation in all cases. Bacillus cereus isolates were sensitive to clindamycin hydrochloride hydrate and aminoglycosides but resistant to penicillins and cephalosporins.

To investigate an outbreak of Burkholderia (formerly Pseudomonas) cepacia respiratory tract colonization and infection in mechanically ventilated patients. A retrospective case-control and bacteriologic study. Veterans Affairs medical center. 42 mechanically ventilated patients who developed respiratory tract colonization or infection with B. cepacia and 135 ventilator-dependent controls who were not colonized and did not develop infections. Clinical and demographic data; benzalkonium chloride concentrations and pH levels in albuterol sulfate solutions; repetitive-element polymerase chain reaction (PCR)-mediated molecular fingerprinting on eight patient isolates and three environmental B. cepacia isolates that were available for study. 42 patients had B. cepacia respiratory tract colonization or infection. Observation of intensive care unit and respiratory care personnel showed faulty infection control procedures (for example, the same multiple-dose bottle of albuterol was used for many mechanically ventilated patients). More case patients (39 [92.9%]) than controls (95 [70.4%]; P = 0.006) received nebulized albuterol, and case patients (67.5 treatments) received more treatments than controls (18 treatments; P < 0.001). In-use albuterol solutions had pH values that were unstable, and benzalkonium chloride concentrations declined over time to levels capable of supporting bacterial growth. Medication nebulizers and in-use bottles of albuterol harbored B. cepacia. Molecular fingerprints of patient isolates and environmental B. cepacia isolates were identical using repetitive-element PCR. No further isolates of B. cepacia were identified after institution of appropriate infection control procedures. Multiple-dose medications and reliance on benzalkonium chloride as a medication preservative provide a mechanism for nosocomial spread of microorganisms, particularly if infection control procedures are not carefully followed. Repetitive-element PCR is a useful fingerprinting technique for molecular epidemiologic studies of B. cepacia.

Thoracic actinomycosis can resemble bronchogenic carcinoma in its clinical presentation and radiographic appearance. We report a case of pulmonary actinomycosis caused by Actinomyces meyeri in which hematogenous dissemination caused multiple brain abscesses resembling metastatic lung cancer. The correct diagnosis was made by thin-needle aspiration of a pleura-based lung mass. The pathogen isolated was further identified with the use of 16S rDNA sequencing. Antibiotic therapy resulted in rapid improvement of the lung lesion; however, the brain lesions required surgical drainage. Antibiotics were continued for more than a year before magnetic resonance images showed complete resolution of the cerebral abscesses.