Identification and characterization of breakthrough contaminants associated with the conventional isolation of Mycobacterium tuberculosis.

Breakthrough contamination of tuberculosis (TB) cultures is a problem in that it allows the overgrowth of another bacterium present in the sputum specimen, which can potentially mask the presence of Mycobacterium tuberculosis. The aim of this study was to isolate and characterize the bacterial organisms responsible for such overgrowth and contamination, and to examine their susceptibility to (i) various chemical selective decontamination steps and (ii) antibiotics in liquid culture media, in an attempt to develop a method to help alleviate contamination problems associated with the conventional isolation of M. tuberculosis from routine patient sputum specimens. Bacterial contaminants from 102 routine sputum cultures were identified molecularly by 16S rRNA gene PCR and direct sequencing from contaminated Löwenstein-Jensen (LJ) slopes and BacT/Alert liquid medium. It was found that the contaminants from LJ slopes belonged to 11 different genera and were composed largely of Gram-negative organisms (84.9 %; 45/53), whereas the liquid culture contaminants belonged to 13 different genera, with 37/66 isolates (56.1 %) being Gram-negative. Pseudomonas aeruginosa was the dominant contaminant in both media. The effect of six different selective decontamination protocols was examined. Four of the six methods were effective at eliminating all culturable organisms present; these were 5 % oxalic acid, 5 % oxalic acid/2 % NaOH, 5 % oxalic acid/4 % NaOH and 1 % chlorhexidine. NaOH at a concentration of 2 or 4 % was less effective as it was unable to eliminate all organisms of each species tested, with the exception of P. aeruginosa. In conclusion, breakthrough contamination of TB cultures is due to a diverse range of at least 17 different bacterial genera, with P. aeruginosa and Staphylococcus epidermidis accounting for the dominant contaminating flora. Employment of chemical decontaminating protocols solely involving NaOH may lead to higher rates of contamination. Where such contamination is encountered, TB laboratories should consider the reprocessing of such sputum samples with an alternative decontamination method such as 1 % chlorhexidine.

Characterization of urease-positive thermophilic Campylobacter subspecies by multilocus enzyme electrophoresis typing.

Thirty-one urease-positive thermophilic Campylobacter (UPTC) isolates, including three reference strains (NCTC12892, NCTC12895 and NCTC12896), and three Campylobacter lari isolates, which were isolated from several countries and sources, were compared genotypically by using multilocus enzyme electrophoresis (MLEE). We examined allelic variation around seven enzyme loci, including the adenylate kinase, alkaline phosphatase, catalase, fumarase, malic enzyme, malate dehydrogenase, and L-phenylalanyl-L-leucine peptidase loci. MLEE typing revealed the presence of 23 different electrophoretic types (ETs) among the 31 UPTC isolates, and 14 isolates shared six electrophoretic profiles. Three different ETs were identified for the three C. lari isolates examined, and no ETs were shared by UPTC and C. lari isolates. Quantitative analyses were subsequently performed by using allelic variation data, and the results demonstrated that the mean genetic diversity was 0.655. In conclusion, MLEE demonstrated that the UPTC isolates examined are genetically hypervariable and form a cluster separate from the C. lari cluster.

Multilocus enzyme electrophoresis: a practical guide.

Multilocus enzyme electrophoresis (MEE) uses the relative electrophoretic mobilities of intracellular enzymes to characterize and differentiate organisms by generating an electromorph type (ET). This article presents the chemical conditions that may be useful, a guide to the successful practice of the electrophoretic technique, and analysis of the results.

Preliminary evaluation of the Arctic Sun temperature-controlling system during off-pump coronary artery bypass surgery.

BACKGROUND: Maintaining normothermia during off-pump coronary artery bypass (OPCAB) surgery is a challenge not met by currently available medical devices and strategies. The purpose of this study was to determine the efficacy of a new thermoregulatory device, the Arctic Sun temperature-controlling circulating fluid adhesive pad system, in preventing hypothermia during OPCAB surgery. METHODS: Thirteen consenting patients undergoing OPCAB had their temperature managed using the Arctic Sun system. They were matched with 23 consenting control OPCAB patients whose temperature was maintained with standardized, conventional therapy (elevated ambient operating room temperature, warmed intravenous fluids, and a convective forced air warming system placed under the surgical drapes). Nasopharyngeal temperature (recorded at 1-minute intervals) was compared between the two groups by analysis of both the time and area under the curve for a temperature less than 36 degrees C. RESULTS: Multivariate linear regression analysis revealed that the average amount of hypothermia in the Arctic Sun group was significantly less than in the control group, both for time spent less than 36 degrees C (20.7 vs 121.3 minutes, p = 0.0004) and for area under the curve less than 36 degrees C (11.8 degrees C vs 78.1 degrees C x minutes, p = 0.0001). CONCLUSIONS: The Arctic Sun temperature-controlling system is more effective than conventional warming methods in preventing hypothermia during OPCAB surgery.

The impact of postoperative atrial fibrillation on neurocognitive outcome after coronary artery bypass graft surgery.

UNLABELLED: Neurocognitive decline is a continuing source of morbidity after cardiac surgery. Atrial fibrillation occurs often after cardiac surgery and has been linked to adverse neurologic events. We sought to determine whether postoperative atrial fibrillation was associated with postoperative cognitive dysfunction. Four-hundred-eleven patients were enrolled to receive a battery of neurocognitive tests both preoperatively and 6 wk after elective coronary artery bypass graft surgery. Neurocognitive test scores were separated into four cognitive domains, with a composite cognitive index (the mean of the four domain scores) determined for each patient at every testing period. Multivariable analysis controlling for age, years of education, diabetes mellitus, left ventricular ejection fraction, and preoperative atrial fibrillation compared the presence of postoperative atrial fibrillation with change in cognitive function. Three-hundred-eight patients completed both pre- and postoperative cognitive testing; 69 patients (22%) had postoperative atrial fibrillation. Those who developed atrial fibrillation showed more cognitive decline than those who did not develop postoperative atrial fibrillation (P = 0.036). Atrial fibrillation was associated with poorer cognitive function 6 wk after surgery. Although the mechanism of this association is yet to be determined, prevention of atrial fibrillation may result in improved neurocognitive function. IMPLICATIONS: Neurocognitive dysfunction is common after coronary artery bypass graft surgery. The relationship between atrial fibrillation and neurocognitive dysfunction has not been examined. Our study shows that postoperative atrial fibrillation is associated with neurocognitive decline.

The rewarming rate and increased peak temperature alter neurocognitive outcome after cardiac surgery.

UNLABELLED: Neurocognitive dysfunction is a common complication after cardiac surgery. We evaluated in this prospective study the effect of rewarming rate on neurocognitive outcome after hypothermic cardiopulmonary bypass (CPB). After IRB approval and informed consent, 165 coronary artery bypass graft surgery patients were studied. Patients received similar surgical and anesthetic management until rewarming from hypothermic (28 degrees -32 degrees C) CPB. Group 1 (control; n = 100) was warmed in a conventional manner (4 degrees -6 degrees C gradient between nasopharyngeal and CPB perfusate temperature) whereas Group 2 (slow rewarm; n = 65) was warmed at a slower rate, maintaining no more than 2 degrees C difference between nasopharyngeal and CPB perfusate temperature. Neurocognitive function was assessed at baseline and 6 wk after coronary artery bypass graft surgery. Univariable analysis revealed no significant differences between the Control and Slow Rewarming groups in the stroke rate. Multivariable linear regression analysis, examining treatment group, diabetes, baseline cognitive function, and cross-clamp time revealed a significant association between change in cognitive function and rate of rewarming (P = 0.05). IMPLICATIONS: Slower rewarming during cardiopulmonary bypass (CPB) was associated with better cognitive performance at 6 wk. These results suggest that a slower rewarming rate with lower peak temperatures during CPB may be an important factor in the prevention of neurocognitive decline after hypothermic CPB.