Four-year prospective evaluation of community-acquired bacteremia: epidemiology, microbiology, and patient outcome.

The objectives of this study were to (1) describe the epidemiology and microbiology of community-acquired bacteremia; (2) determine the crude mortality associated with such infections; and (3) identify independent predictors of mortality. All patients with clinically significant community-acquired bacteremia admitted to a university-affiliated Veterans Affairs medical center from January 1994 through December 1997 were evaluated. During the study period, 387 bacteremic episodes occurred in 334 patients. Staphylococcus aureus, Escherichia coli, and coagulase-negative staphylococci were the most commonly isolated organisms; the most frequent sources were the urinary tract and intravascular catheters. Approximately 14% of patients died. Patient characteristics independently associated with increased mortality included shock (OR 3.7, p = 0.02) and renal failure (OR 4.0, p = 0.003). The risk of death was also higher in those whose source was pneumonia (OR 6.3, p = 0.03) or an intra-abdominal site (OR 10.7, p = 0.02), or if multiple sources were identified (OR 13.4, p = 0.003). Community-acquired bacteremia is often device-related and may be preventable. Strategies that have been successful in preventing nosocomial device-related bacteremia should be adapted to the outpatient setting.

Four year prospective evaluation of nosocomial bacteremia: epidemiology, microbiology, and patient outcome.

A prospective study of all patients with clinically significant nosocomial bacteremia at one institution from 1994 to 1997 was performed to: (1) describe the epidemiology and microbiology of nosocomial bacteremias; (2) determine the crude mortality associated with such infections; and (3) identify independent predictors of mortality. Four hundred four episodes of bacteremia occurred in 322 patients; the crude in-hospital mortality was 31%. Coagulase-negative staphylococci, Staphylococcus aureus, and enterococci were the leading pathogens, and intravascular catheters were the most frequently identified source. The highest mortality occurred in patients with candidemia (67%). Independent predictors of mortality included evidence of shock at the time of infection, acquisition of bacteremia in an intensive care unit, a "Do Not Attempt Resuscitation" order, and the presence of certain comorbid conditions (e.g., malignancy, HIV infection). Because many of these infections may be preventable, education of health care providers and strict adherence to established infection control practices are critical.

Catheter-directed thrombolysis in the treatment of phlegmasia cerulea dolens.

Phlegmasia cerulea dolens is a potentially devastating complication of extensive deep venous thrombosis for which there is currently no consensus for treatment. Heparin anticoagulation, surgical thrombectomy, thrombolytic therapy, fasciotomy, and amputation have each been advocated. We present two cases of phlegmasia cerulea dolens successfully treated with catheter-directed venous thrombolytic therapy.

Clinical application of the Mycobacterium tuberculosis direct test: case report, literature review, and proposed clinical algorithm.

The relatively new Mycobacterium tuberculosis direct test (MTDT) enzymatically amplifies M tuberculosis complex 16s ribosomal RNA. The sensitivity of the test ranges from 75 to 100%, with specificity of 95 to 100%, positive predictive value between 78% and 100%, and negative predictive value between 95% and 100%. Similar test characteristics have been documented in nonrespiratory specimens and in specimens that ultimately grow nontuberculous mycobacterium (NTM). This test allows for rapid identification of M tuberculosis in the smear-positive patient and may greatly improve sensitivity over acid-fast bacilli smear alone. A negative test result with a positive smear suggests infection with NTM or Mycobacterium avium complex. We present a case that illustrates the value of MTDT for analysis of tissue specimens in immunocompromised patients with suspected mycobacterial disease and review the rapidly developing literature about this test. We propose an algorithm using MTDT, acid-fast smear, and mycobacterial culture for the diagnosis and treatment of the immunocompromised patient with suspected mycobacterial infection.

Quantitative analysis of respiratory, motor, and sensory function after supraclavicular block.

UNLABELLED: The incidence and clinical significance of hemidiaphragmatic paresis after supraclavicular block of the brachial plexus is unknown. Eight healthy volunteers received a supraclavicular block with a standard technique using 30 mL of 1.5% lidocaine. Respiratory function was assessed with ultrasound of the diaphragm, respiratory inductive plethysmography (RIP), and pulmonary function tests (PFT) every 20 min. Sensory block was assessed with pinprick and motor block with isometric force dynamometry every 20 min. Four of eight subjects demonstrated hemidiaphragmatic paresis on both ultrasound and RIP. No subject experienced changes in PFT values or subjective symptoms of respiratory difficulty. Motor and sensory blockade outlasted hemidiaphragmatic paresis. These results are contrasted to the often symptomatic, 100% incidence of hemidiaphragmatic paresis seen after interscalene block. In this study of healthy volunteers, supraclavicular block was associated with a 50% incidence (95% confidence interval 14-86) of hemidiaphragmatic paresis that was not accompanied by clinical evidence of respiratory compromise. IMPLICATIONS: Interscalene block is always associated with diaphragmatic paralysis and respiratory compromise. The significance of these side effects after supraclavicular block is unknown. Using sensitive measures of respiratory function, we determined that diaphragmatic paralysis occurs less often with the supraclavicular approach and is not associated with respiratory difficulties in healthy subjects.

Detection of immunoglobulin G (IgG) and IgM antibodies to Toxoplasma gondii: evaluation of four commercial immunoassay systems.

A comparative evaluation of the following commercial immunoassays for the determination of antibodies to Toxoplasma gondii was performed: Behring Diagnostics OPUS Toxo G and Toxo M, Abbott Diagnostics IMX Toxo-IgG 2.0 and Toxo-IgM, Sanofi Diagnostics Pasteur Platelia Toxo IgG and Toxo IgM, and bioMérieux Vitek VIDAS Toxo IgG and IgM. Of 676 specimens that were tested for Toxoplasma-specific immunoglobulin G (IgG) antibodies, 26% were reactive by all methods while 8% displayed some discrepancy. Of 718 specimens that were tested for Toxoplasma-specific IgM antibodies, 3% were reactive by all methods while 10% displayed some discrepancy. Analysis of discrepant specimens revealed performance shortcomings with all IgM-specific assays. The impact of such shortcomings is magnified in a population with a low prevalence of toxoplasmosis.

Transverse colon volvulus and associated Chilaiditi's syndrome: case report and literature review.

Chilaiditi's sign is a radiographic term used when the hepatic flexure of the colon is seen interposed between the liver and right hemidiaphragm. When symptomatic, this is Chilaiditi's syndrome. We report a case of Chilaiditi's syndrome associated with transverse colon volvulus. A 64-yr-old male presented with abdominal pain, vomiting, and constipation. Barium enema demonstrated complete colonic obstruction, leading to operative decompression and right hemicolectomy. This is the second case in the English literature of transverse colon volvulus associated with Chilaiditi's syndrome. Colonic elongation and laxity of colonic and hepatic suspensory ligaments are the principal predisposing factors to these two entities.

Use of the fluorochrome calcofluor white in the screening of stool specimens for spores of microsporidia.

Microsporidia are obligate intracellular protozoal pathogens associated with chronic diarrhea in individuals infected with HIV. Direct detection methods for microsporidial spores in stool include chromotrope-based, fluorochrome, and immunofluorescent stains. The authors compared the ability to detect microsporidial spores in 168 stool specimens using two stains: a chromotrope-based modified trichrome stain and a fluorochrome stain, calcofluor white (Cellufluor, Polysciences, Warrington, PA). In addition to being faster and easier to perform, the calcofluor white stain was found to be more sensitive than the chromotrope-based stain, as 6 of 24 specimens positive by calcofluor white were negative by the chromotrope-based stain on initial smear evaluation. Repeat examination confirmed these six as being positive. To evaluate the specificity of the calcofluor white stain, 20 formalin-fixed stool specimens (5 positive and 15 negative for microsporidial spores) were evaluated in blinded fashion by two affiliated clinical laboratories using their own formulations of calcofluor white. A single discrepant result (falsely positive) was reported from one laboratory. The use of the calcofluor white stain is recommended as a simple and highly sensitive screening procedure for the detection of microsporidial spores in stool specimens.

Rapid horizontal gaze movement in the monkey.

1. We studied horizontal eye and head movements in three monkeys that were trained to direct their gaze (eye position in space) toward jumping targets while their heads were both fixed and free to rotate about a vertical axis. We considered all gaze movements that traveled > or = 80% of the distance to the new visual target. 2. The relative contributions and metrics of eye and head movements to the gaze shift varied considerably from animal to animal and even within animals. Head movements could be initiated early or late and could be large or small. The eye movements of some monkeys showed a consistent decrease in velocity as the head accelerated, whereas others did not. Although all gaze shifts were hypometric, they were more hypometric in some monkeys than in others. Nevertheless, certain features of the gaze shift were identifiable in all monkeys. To identify those we analyzed gaze, eye in head position, and head position, and their velocities at three points in time during the gaze shift: 1) when the eye had completed its initial rotation toward the target, 2) when the initial gaze shift had landed, and 3) when the head movement was finished. 3. For small gaze shifts (< 20 degrees) the initial gaze movement consisted entirely of an eye movement because the head did not move. As gaze shifts became larger, the eye movement contribution saturated at approximately 30 degrees and the head movement contributed increasingly to the initial gaze movement. For the largest gaze shifts, the eye usually began counterrolling or remained stable in the orbit before gaze landed. During the interval between eye and gaze end, the head alone carried gaze to completion. Finally, when the head movement landed, it was almost aimed at the target and the eye had returned to within 10 +/- 7 degrees, mean +/- SD, of straight ahead. Between the end of the gaze shift and the end of the head movement, gaze remained stable in space or a small correction saccade occurred. 4. Gaze movements < 20 degrees landed accurately on target whether the head was fixed or free. For larger target movements, both head-free and head-fixed gaze shifts became increasingly hypometric. Head-free gaze shifts were more accurate, on average, but also more variable. This suggests that gaze is controlled in a different way with the head free. For target amplitudes < 60 degrees, head position was hypometric but the error was rather constant at approximately 10 degrees.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of volume and periodicity on blood cultures.

Blood specimens collected for culture by using the high-volume resin-based BACTEC system over an 18-month period at the Seattle Veterans Administration Center were examined in this study. Of 7,783 cultures obtained, 624 were classified as true positives. Patients in this group had between 20 and 60 ml of blood drawn per culture and separated into 10-ml aliquots for incubation. Analysis of the results stratified by cultured volume and time interval between specimen collection accorded yield advantage to culture volume at the maximal amounts tested. No advantage was observed with any particular interval of collection. Increasing cultured volume from 20 to 40 ml increased yield by 19%. Increasing cultured volume from 40 to 60 ml increased yield by an additional 10%. The same effect was seen whether cultures were drawn simultaneously or serially within 24 h. These observations support other reports demonstrating increased yield with increased cultured blood volume. However, they demonstrate increases in yield at volumes much higher than previously considered. In conclusion, this study demonstrates that high-volume blood cultures drawn serially or simultaneously return the best yields.

Invasive Haemophilus influenzae infections in older children and adults in Seattle.

We recently saw two unusual manifestations of Haemophilus influenzae infection in adults in the Seattle area: fulminant sepsis in an otherwise-healthy man and three episodes of bacteremia in a woman with chronic liver disease. We retrospectively identified 79 bacteremic and 40 non-bacteremic cases of invasive H. influenzae infection developing in patients > or = 9 years of age between 1 January 1980 and 31 December 1990. The most common clinical presentations among patients with bacteremia included pneumonia (52%), septicemia (27%), meningitis (8%), gynecologic infection (5%), and epiglottitis (5%). Underlying illnesses were common in these patients, and overall mortality was 35.5%. Factors associated with mortality included underlying neurological disease, polymicrobial bacteremia, and advanced age. The clinical presentations of the 40 patients without bacteremia included soft-tissue abscesses (45%), lung abscesses (18%), peritonitis (13%), meningitis (8%), gynecologic infection (8%), epididymitis (5%), mastoiditis (3%), and osteomyelitis (3%). Thus H. influenzae disease has a variety of presentations and is associated with significant mortality in older children and adults. Further study is required to determine whether widespread administration of H. influenzae type b conjugate vaccine to infants will alter the development of subsequent disease in later life.

A survey of clinical isolates of Enterobacteriaceae using a series of DNA probes for aminoglycoside resistance genes.

DNA probes specific for the aminoglycoside adenylyltransferase ANT(2"), the aminoglycoside phosphotransferases APH(3')-I and APH(3')-II, and the aminoglycoside acetyltransferases AAC(3)-I, AAC(3)-V and AAC(6')-I were used to screen 151 clinical isolates for the presence of these resistance determinants. The two most common resistance genes in the study isolates were those encoding ANT(2") and APH(3')-I. The phenotypic pattern of aminoglycoside resistance, as determined by a modified disk diffusion test correlated well with the genotypes of the organisms as established using DNA probes, although strains carrying the ANT(2") gene could express one of several phenotypes.

Evaluation of four mycobacterial blood culture media: BACTEC 13A, Isolator/BACTEC 12B, Isolator/Middlebrook agar, and a biphasic medium.

Four commercially available mycobacterial blood culture systems were compared for sensitivity and time to detection of growth. A 5-ml volume of SPS-anticoagulated blood was cultured in a BACTEC 13A vial and a modified M7H11/BHI biphasic medium. In addition, two aliquots of Isolator concentrates, each derived from 5 ml of blood, were inoculated into a BACTEC 12B vial and onto a pair of Middlebrook 7H11 agar plates (M7H11). Mycobacteria were recovered from 32 of 180 cultured specimens (17.8%). Growth was detected in 30 (93.7%) of the 13A vials, 27 (84.4%) of the M7H11 agar plates, 26 (81.2%) of the 12B vials, and 14 (43.8%) of the biphasic bottles. The mean times to growth detection in the 13A vial (14.2 days) and the 12B vial (13.7 days) were shorter than in either the M7H11 plates (20.8 days) or the biphasic medium (24.1 days). When the Isolator/12B vial-and-M7H11 plates were evaluated as a single system, 29 cultures (90.6%) had a mean time to growth detection of 13.5 days. Colony-forming units per ml were inversely associated with time to growth detection. Delay in transport (greater than 24 h) appeared to reduce viability. The direct inoculation feature makes the 13A vial very suitable for mycobacterial blood cultures.

Automated antibiotic susceptibility testing: comparative evaluation of four commercial systems and present state.

Automation of AST has come quite some way and is here to stay. In particular, fully automated, "hands off" instruments have great appeal to laboratories with a limited number of well-trained and experienced clinical microbiology personnel. None of the evaluated instruments is perfect, but then neither are the standard or reference techniques. Overnight incubation has been the yardstick since the early days of in vitro AST. Given the usually shorter therapeutic intervals of 4- to 12-hour dosage schedules, it is quite possible that shorter incubation times for in vitro tests will become more of a standard. Until that time, newer, including automatic, techniques need to be evaluated against the more traditional standard methods. Quality control is critical, and since no systematic approach aside from individual manufacturers' suggestions exists, it should be developed by the NCCLS or similar agencies. Quality control might include standards for the evaluation of such equipment and systems because the development of new technology in this area will continue. Overall, reproducibility and accuracy of the instruments and methods evaluated were quite promising and should encourage well-designed studies of clinical correlation and relevance. The AMS equipment has been in use for routine AST in the clinical laboratories of the Seattle Veterans Administration Medical Center and the University of Washington Hospital. Because of its simplicity and flexibility, the Kirby-Bauer method continues to be an alternate technique for certain important clinical isolates, for instance, blood cultures in both laboratories. Finally, it should be remembered that the most critical function of all such equipment is the reliable detection of resistance.

Development of a DNA probe from the deoxyribonucleotide sequence of a 3-N-aminoglycoside acetyltransferase [AAC(3)-I] resistance gene.

The aacC1 gene encoding the 3-N-aminoglycoside acetyltransferase [AAC(3)-I] was cloned from enteric plasmid pJR88, and its deoxyribonucleotide sequence was determined. Significant nucleotide homology was noted in the region extending from the proposed -35 sequences through the first 59 base pairs of the aacC1 gene open reading frame (ORF) and the upstream flanking regions and ORFs of several other antibiotic resistance genes. Sequences were noted to be homologous with the 6'-N-aminoglycoside acetyltransferase [AAC(6')-I], 2''-O-aminoglycoside adenylyltransferase [AAD(2'')], and 3''-O-aminoglycoside adenylyltransferase [AAD(3'')] resistance genes; the OXA-1, OXA-2, and PSE-2 beta-lactamase genes; and several dihydrofolate reductase genes. Small regions of homology were noted in the 3'-flanking regions of these resistance genes as well. A DNA probe for the aacC1 gene was selected from the nucleotide sequence information and was tested against a series of genetically and enzymatically defined strains. The probe, which proved specific for the aacC1 gene, was then tested against a series of 58 gentamicin-susceptible and 219 gentamicin-resistant gram-negative bacilli isolated from patients at the Seattle Veterans Administration Medical Center. Only six clinical isolates were noted to carry the aacC1 gene. Each was resistant to gentamicin but susceptible to kanamycin, tobramycin, and amikacin. The presence of homologous regions of DNA at both the 3' and 5' ends of the aacC1 gene reinforces the importance of choosing probes from within the ORFs of genes and of avoiding flanking sequences. When the homology with other sequences extends into the ORF, as it does with the aacC1 gene, development of a specific probe may require determination of the nucleotide sequence.

Cloning and sequencing of a gene encoding an aminoglycoside 6'-N-acetyltransferase from an R factor of Citrobacter diversus.

The aacA1 gene, which encodes a 6'-N-acetyltransferase [AAC(6')-I] mediating resistance to kanamycin, tobramycin, and amikacin, was cloned from the Citrobacter diversus R plasmid pBWH100 into the Escherichia coli vector pBR322. The complete nucleotide sequence of the gene and flanking regions was determined. A protein of approximately 21 kilodaltons was identified when the chimeric plasmid encoding the aacA1 gene was introduced into E. coli maxicells. This value is consistent with the size predicted for a protein translated from the open reading frame of the gene.