Comparison of BD Bactec Plus Aerobic/F medium to VersaTREK Redox 1 blood culture medium for detection of Candida spp. in seeded blood culture specimens containing therapeutic levels of antifungal agents

Recovery of Candida spp. using the BD Bactec FX blood culture (BC) system (Bactec Plus Aerobic/F medium) and the VersaTREK system (aerobic Redox medium) was evaluated using seeded BC bottles with and without the addition of commonly used antifungal agents. BC bottles (n = 1,442) were each inoculated with 10 ml human whole blood and 0.1 ml of suspensions of Candida spp., with or without antifungal agents. BC bottles were incubated in the corresponding system for a maximum of 5 days. In the absence of antifungal agents, Bactec FX recovered 97.4% of Candida spp., and VersaTREK recovered 99.1% (P = 0.154). With regard to length of time to detection (LTD) and overall recovery, both systems had various levels of effectiveness in recovering C. glabrata. In bottles containing antifungal agents, Bactec FX recovered 83.1% of isolates, whereas VersaTREK recovered 50.7% of Candida spp. (P < 0.001). For BC bottles without the addition of antifungal agents, the median LTD for VersaTREK was 2.2 h faster than that of Bactec FX (P < 0.001). In the presence of antifungal agents, the Bactec FX recovery time was significantly faster than that of VersaTREK (median difference of 10.8 h, P < 0.001). We conclude that both systems have comparable abilities to recover Candida spp. from seeded blood cultures in the absence of antifungal agents. In the presence of therapeutic levels of commonly used antifungal agents, the Bactec FX system demonstrated a significantly greater recovery of various Candida spp., as well as a shorter LTD.     

Superior Sensitivity and Decreased Time to Detection with the Bactec Peds Plus/F System Compared to the BacT/Alert Pediatric FAN Blood Culture System

Here, we compare the sensitivities and times to detection (TTD) of BacT/Alert Pediatric FAN (PF) and Bactec Peds Plus blood culture bottles. Test bottles were inoculated with 2 ml of banked whole blood, 1-ml aliquots of antibiotic suspension, and organisms diluted to simulate a bacteremia level of 10 to 100 CFU/ml. The control bottles were inoculated with 3 ml of banked blood and organism suspensions only. The organism-drug combinations were Staphylococcus epidermidis and vancomycin, methicillin-resistant Staphylococcus aureus and vancomycin, Streptococcus pneumoniae, vancomycin, and ceftriaxone, Streptococcus agalactiae, ampicillin, and cefotaxime, Escherichia coli, cefotaxime, and cefepime, Pseudomonas aeruginosa, piperacillin-tazobactam, cefepime, and gentamicin, Neisseria meningitidis and ceftriaxone, and Haemophilus influenzae and ceftriaxone. The control and test bottle combinations were tested in duplicate. The bottles were incubated for 5 days; 32 control and 104 test bottles were incubated. Overall, the bacterial recovery rates for the PF and Peds Plus bottles were 37% and 62%, 94% and 100% in the controls, 19% and 50% in the test bottles, and 33% and 92% in the bottles with vancomycin, respectively. No bacteria were recovered from the bottles with S. pneumoniae, S. agalactiae, E. coli, N. meningitidis, or H. influenzae in combination with cefotaxime or ceftriaxone. The Peds Plus system detected P. aeruginosa in bottles with cefepime and piperacillin-tazobactam, but the PF system recovered bacteria only in bottles with trough levels of piperacillin-tazobactam. The mean TTD were shorter in the Peds Plus system controls (14.2 versus 18.0 h; P = 0.001) and the test bottles (14.3 versus 17.8 h; P = 0.008) than in the PF bottles. Overall, we demonstrated superior sensitivity, TTD, and antibiotic neutralization in the Bactec Peds Plus system compared to those in the Pediatric FAN system.     

Sonication of Explanted Prosthesis Combined with Incubation in BD Bactec Bottles for Pathogen-Based Diagnosis of Prosthetic Joint Infection

Prosthetic joint infection (PJI) is a rare but refractory complication of arthroplasty. Accurate identification of pathogens is a key step for successful treatment of PJI, which remains a challenge for clinicians and laboratory workers. We designed a combined culture method with sonication of implants and incubation in a BD Bactec system to improve the effectiveness of pathogen diagnosis in PJI. The aims of this study were to investigate the diagnostic accuracy of sonicate fluid cultures in the BD Bactec system and to compare the results with those of synovial fluid cultures in the BD Bactec system. The prosthetic components removed were sonicated in Ringer''s solution, and then sonicate fluid was incubated in Bactec bottles for 5 days. Synovial fluid was incubated in Bactec bottles for 5 days as a control. Synovial fluid cultures with Bactec bottles and sonicate fluid cultures with Bactec bottles showed sensitivities of 64% and 88%, respectively (P = 0.009), with specificities of 98% and 87% (P = 0.032), respectively. Sonicate fluid cultures with Bactec bottles were more sensitive than synovial fluid cultures with Bactec bottles regardless of whether antimicrobial agents were used within 14 days before surgery (81% versus 52%; P = 0.031) or not (93% versus 72%; P = 0.031). Sonication of explanted prostheses followed by incubation of the resulting sonicate fluid in Bactec bottles detected many more pathogens than did synovial fluid cultures with Bactec bottles. This method is also effective in cases with antibiotic treatment before surgery.     

Use of the BacT/Alert Blood Culture System for Culture of Sterile Body Fluids Other than Blood

Studies have demonstrated that large-volume culture methods for sterile body fluids other than blood increase recovery compared to traditional plated-medium methods. BacT/Alert is a fully automated blood culture system for detecting bacteremia and fungemia. In this study, we compared culture in BacT/Alert standard aerobic and anaerobic bottles, BacT/Alert FAN aerobic and FAN anaerobic bottles, and culture on routine media for six specimen types, i.e., continuous ambulatory peritoneal dialysate (CAPD), peritoneal, amniotic, pericardial, synovial, and pleural fluids. Specimen volumes were divided equally among the three arms of the study. A total of 1,157 specimens were tested, with 227 significant isolates recovered from 193 specimens. Recovery by method was as follows: standard bottles, 186 of 227 (82%); FAN bottles, 217 of 227 (96%); and routine culture, 184 of 227 (81%). The FAN bottles recovered significantly more gram-positive cocci (P < 0.001), Staphylococcus aureus (P = 0.003), coagulase-negative staphylococci (P = 0.008), gram-negative bacilli (P < 0.001), Enterobacteriaceae (P = 0.005), and total organisms (P < 0.001) than the routine culture. There were no significant differences in recovery between the standard bottles and the routine culture. The FAN aerobic bottle recovered significantly more gram-positive cocci (P < 0.001), S. aureus isolates (P < 0.001), coagulase-negative staphyococci (P = 0.003), and total organisms (P < 0.001) than the standard aerobic bottle, while the FAN anaerobic bottle recovered significantly more gram-positive cocci (P < 0.001), S. aureus isolates (P < 0.001), Enterobacteriaceae (P = 0.03), and total organisms (P < 0.001) than the standard anaerobic bottle. For specific specimen types, significantly more isolates were recovered from the FAN bottles compared to the routine culture for synovial (P < 0.001) and CAPD (P = 0.004) fluids. Overall, the FAN bottles were superior in performance to both the standard bottles and the routine culture for detection of microorganisms from the types of sterile body fluids included in this study.     

Clinical Evaluation of BacT/Alert FA Plus and FN Plus Bottles Compared with Standard Bottles

The performance of the BacT/Alert FA Plus and FN Plus resin bottles was evaluated in comparison with that of standard aerobic (SA) and standard anaerobic (SN) bottles. Twenty milliliters of blood from adult patients was equally distributed into four types of bottles: FA Plus, FN Plus, SA, and SN. The detection of clinically significant organisms and the time to detection (TTD) were monitored for each bottle. Among the 3,103 blood culture sets that were requested, the blood volume of each bottle was over 4 ml in 1,481 sets (47.7%). Among these 1,481 sets, 158 cultures grew in the FA Plus and SA bottles, and 136 grew in the FN Plus and SN bottles. Growth in only one type of bottle was more commonly observed for the FA Plus (n = 38) than for the SA (n = 14) (P = 0.001) bottles and for the FN Plus (n = 27) than for the SN (n = 10) (P = 0.008) bottles. Gram-negative bacilli were more frequently isolated in the resin bottles (P < 0.05). The skin contamination rate was 1.2% in the resin bottles and the standard bottles. The mean TTD was 11.1 h in the FA Plus bottles versus 13.1 h in the SA bottles (P < 0.001) and 12.0 h in the FN Plus bottles versus 12.8 h in the SN bottles (P = 0.083). Clinically significant bacteria, including Gram-negative bacilli, were isolated more frequently from the resin bottles than from the standard bottles. Clinically significant bacteria were detected faster using the aerobic resin bottles than using the standard aerobic bottles. This finding might not be applicable to the standard-practice 10-ml protocol for each bottle because the results from using a smaller volume (5 ml) might be less pronounced.     

Isolation of Kingella kingae from Synovial Fluids Using Four Commercial Blood Culture Bottles

 According to the literature, Kingella kingae may be an underdiagnosed cause of joint and bone infections in children. The use of the Bactec blood culture system for culture of joint fluids has dramatically improved the isolation of this fastidious bacterium. The aim of this study was to test the recovery rate and detection time of four commercial blood culture systems: three different BacT/Alert (Organon Teknika, USA) bottles and one Bactec (Becton Dickinson Microbiology Systems, USA) bottle, all inoculated with Kingella kingae strains mixed with pooled synovial fluids. For each strain the same inoculum and volume of synovial fluid was distributed into each of the four bottles. All 24 strains tested grew in the BacT/Alert Aerobic (100%) and the BacT/Alert Pedi-BacT (100%) bottles. Twenty-one strains grew in the BacT/Alert FAN aerobic (88%) bottle, and 15 strains grew in the Bactec Plus Aerobic F (63%) bottle, in both systems within 12 days (P<0.01). The Kingella kingae strains were first detected in the BacT/Alert Pedi-BacT bottles (P<0.001). The results were reproducible. The BacT/Alert blood culture bottles were superior to previously described blood culture systems in isolating Kingella kingae from synovial fluid, even with small inoculums and small volumes of synovial fluid.     

A Comprehensive Evaluation of the Bruker Biotyper MS and Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Yeasts,Part of the National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) Study, 2012 to 2013

Among the 2,683 yeast isolates representing 41 different species (25 Candida and Candida-related species and 16 non-Candida yeast species) collected in the National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) program (2012 to 2013), the Bruker Biotyper MS matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system exhibited significantly higher accuracy rates than the Vitek MS system for identification of all yeast isolates (98.8% versus 95.4%, P <0.001 by Pearson''s chi-square test) and for all Candida and Candida-related species isolates (99.4% versus 95.5%, P < 0.001).     

Multicenter Evaluation of Candida QuickFISH BC for Identification of Candida Species Directly from Blood Culture Bottles

Candida species are common causes of bloodstream infections (BSI), with high mortality. Four species cause >90% of Candida BSI: C. albicans, C. glabrata, C. parapsilosis, and C. tropicalis. Differentiation of Candida spp. is important because of differences in virulence and antimicrobial susceptibility. Candida QuickFISH BC, a multicolor, qualitative nucleic acid hybridization assay for the identification of C. albicans (green fluorescence), C. glabrata (red fluorescence), and C. parapsilosis (yellow fluorescence), was tested on Bactec and BacT/Alert blood culture bottles which signaled positive on automated blood culture devices and were positive for yeast by Gram stain at seven study sites. The results were compared to conventional identification. A total of 419 yeast-positive blood culture bottles were studied, consisting of 258 clinical samples (89 C. glabrata, 79 C. albicans, 23 C. parapsilosis, 18 C. tropicalis, and 49 other species) and 161 contrived samples inoculated with clinical isolates (40 C. glabrata, 46 C. albicans, 36 C. parapsilosis, 19 C. tropicalis, and 20 other species). A total of 415 samples contained a single fungal species, with C. glabrata (n = 129; 30.8%) being the most common isolate, followed by C. albicans (n = 125; 29.8%), C. parapsilosis (n = 59; 14.1%), C. tropicalis (n = 37; 8.8%), and C. krusei (n = 17; 4.1%). The overall agreement (with range for the three major Candida species) between the two methods was 99.3% (98.3 to 100%), with a sensitivity of 99.7% (98.3 to 100%) and a specificity of 98.0% (99.4 to 100%). This study showed that Candida QuickFISH BC is a rapid and accurate method for identifying C. albicans, C. glabrata, and C. parapsilosis, the three most common Candida species causing BSI, directly from blood culture bottles.     

Rapid Antifungal Susceptibility Determination for Yeast Isolates by Use of Etest Performed Directly on Blood Samples from Patients with Fungemia

We prospectively determined the antifungal susceptibility of yeast isolates causing fungemia using the Etest on direct blood samples (195 prospectively collected and 133 laboratory prepared). We compared the Etest direct (24 h of incubation) with CLSI M27-A3 and the standard Etest methodologies for fluconazole, voriconazole, posaconazole, isavuconazole, caspofungin, and amphotericin B. Strains were classified as susceptible, resistant, or nonsusceptible using CLSI breakpoints (voriconazole breakpoints were used for posaconazole and isavuconazole). Categorical errors between Etest direct and CLSI M27-A3 for azoles were mostly minor. No errors were detected for caspofungin, and high percentages of major errors were detected for amphotericin B. For the azoles, false susceptibility (very major errors) was found in only two (0.6%) isolates (Candida tropicalis and C. glabrata). False resistance (major errors) was detected in 46 (14%) isolates for the three azoles (in 23 [7%] after excluding posaconazole). Etest direct of posaconazole yielded a higher number of major errors than the remaining azoles, especially for C. glabrata, Candida spp., and other yeasts. Excluding C. glabrata, Candida spp., and other yeasts, the remaining species did not yield major errors. Etest direct for fluconazole, voriconazole, isavuconazole, and caspofungin shows potential as an alternative to the CLSI M27-A3 procedure for performing rapid antifungal susceptibility tests on yeast isolates from patients with fungemia. Etest direct is a useful tool to screen for the presence of azole-resistant and caspofungin-nonsusceptible strains.The incidence of fungemia continues to rise in many institutions throughout the world, and Candida is one of the leading pathogens isolated from blood (15, 28). Amphotericin B and fluconazole have been widely used for the treatment of fungemia. However, newly licensed antifungal agents (voriconazole, posaconazole, and the echinocandins) and other azoles currently under investigation (isavuconazole) have expanded the antifungal armamentarium.The mortality rate of fungemia remains high (30%) and is clearly correlated with delayed initiation of effective antifungal therapy (11, 17). Antifungal therapy is considered inappropriate when it is omitted, when the agent administered has no antifungal activity against the infecting organism, or when its serum concentrations are subtherapeutic.A growing proportion of Candida isolates obtained from blood samples have reduced antifungal susceptibility to fluconazole and other antifungal agents (14, 25). Patients with candidemia caused by Candida strains with high MICs for fluconazole or voriconazole and echinocandins can have a worse prognosis (22-24). Consequently, systematic use of empirical antifungal agents with broad-spectrum in vitro activity has led to considerable increases in the number of adverse events and in the cost of treatment (2).The combination of an increasing number of antifungal-resistant isolates and the cost of the new antifungal agents makes antifungal susceptibility testing a necessity. The reference antifungal susceptibility testing method for yeasts is the Clinical and Laboratory Standards Institute (CLSI; formerly NCCLS) testing standard M27-A3. However, this method requires pure-culture isolates, and results of antifungal susceptibility testing are not available until 48 to 72 h after the isolation of fungi in blood.The Etest performed directly on blood samples may expedite antifungal testing and provide results in 24 h. Our group has previously demonstrated that the Etest performed directly on samples from the lower respiratory tract is a rapid and accurate procedure for antimicrobial susceptibility testing of bacteria in patients with ventilator-associated pneumonia (4, 5). We compared the results of the Etest performed directly on positive blood cultures with yeasts grown in Bactec blood bottles with the results of CLSI M27-A3 in isolates from patients with fungemia and blood samples generated from previously characterized isolates.(This study was partially presented at the 20th Conference of the European Congress of Clinical Microbiology and Infectious Diseases [ECCMID] in Vienna, Austria, 2010 [abstract no. P-838] [13a].)     

Comparison of Difco ESP and Organon Teknika BacT/Alert continuous-monitoring blood culture systems.

The Difco ESP and Organon Teknika BacT/Alert (BTA) systems were evaluated in a clinical study of 5,421 aerobic and 5,035 anaerobic blood cultures. Of 405 clinically significant positive cultures evaluated, 272 grew in both systems, 86 grew in ESP only, and 47 grew in BTA only (P < 0.005). Of 320 organisms detected in aerobic bottles, 208 grew in both systems, 68 grew in ESP only and 45 grew in BTA only (P < 0.05), with Staphylococcus aureus the only organism showing a statistically significant difference. The ESP anaerobic bottle also detected more anaerobes (16 of 17 versus 4 of 17, P < 0.005) and more organisms overall (57 versus 34, P < 0.05). However, with the exception of patients with anaerobic bacteremia (12 of 13 for ESP and 4 of 13 for BTA, P < 0.05), there was no statistical difference in the detection of patient episodes. Average detection time of matched aerobic bottles was 18.3 h for ESP and 22.0 h for BTA (P < 0.001). For matched pairs of anaerobic bottles, the average detection time was faster in the BTA bottles (P < 0.001), because of the growth of facultative organisms. To explore the differences in anaerobic detection more fully, 20 sets of anaerobic bottles were seeded with 12 anaerobic species mixed with human blood. ESP grew more organisms (17 of 20 versus 10 of 20, P < 0.025), and the average time to detection for the 10 paired positive cultures was 21.6 h for ESP and 50.8 h for BTA (P < 0.05). Times for loading and unloading bottles were similar for both systems.     

Controlled Clinical Comparison of BacT/Alert FA Plus and FN Plus Blood Culture Media with BacT/Alert FA and FN Blood Culture Media

New blood culture media containing antibiotic-binding polymeric beads have been developed for the BacT/Alert (bioMérieux, Inc., Durham, NC) blood culture system. To assess the performance of these new media, we compared the new BacT/Alert aerobic medium (FA Plus) with resins to BacT/Alert FA medium with activated charcoal and the new BacT/Alert anaerobic medium (FN Plus) to BacT/Alert FN medium at 3 tertiary care medical centers. Study bottle pairs were inoculated with a target volume of 6 to 10 ml of blood from adults and incubated in the BacT/Alert 3D blood culture instrument. In the FA Plus versus FA comparison, there were 1,507 study pairs. Among 170 isolates causing true bloodstream infections (BSIs), significantly more Staphylococcus aureus (P < 0.001) and total microorganisms (P < 0.01) grew in the FA Plus bottle than in the FA bottle. Fewer coagulase-negative staphylococcal (CoNS) contaminants grew in the FA Plus bottle than in the FA bottle (10 versus 22; P = 0.05). In addition, growth was detected earlier in the FA Plus bottle than in the FA bottle (P < 0.001). In the FN Plus versus FN comparison, there were 2,386 study pairs. Among 201 isolates causing true BSIs, significantly more S. aureus (P < 0.001), CoNS (P < 0.005), and total microorganisms (P < 0.001) grew in the FN Plus bottle than in the FN bottle. Also, significantly more CoNS contaminants grew in the FN Plus bottle than in the FN bottle (P < 0.001). Overall, microorganisms were detected earlier in the FN Plus than in the FN bottle (P < 0.001). Medical technologists at all 3 study sites preferred the new media for Gram stain interpretation. We conclude that the FA Plus and FN Plus media provide improved and earlier detection of microorganisms compared with the FA and FN media and are preferable for Gram stain interpretation as well.     

Comparison of the two blood culture systems, Bactec 9240 and BacT/Alert 3D, in the detection of Candida spp. and bacteria with polymicrobial sepsis

The purpose of this investigation was to evaluate the performance of the Bactec 9240 and BacT/Alert 3D blood culture systems in the detection of Candida spp. and bacteria in simulated polymicrobial sepsis models. A total of 28 clinical isolates of Escherichia coli, Staphylococcus aureus, Candida albicans, and Candida glabrata were studied. Five polymicrobial models of C. albicans + S. aureus, C. albicans + E. coli, C. glabrata + S. aureus, C. glabrata + E. coli, and C. albicans + C. glabrata were prepared. Each combination was inoculated in five different blood culture vials. The two systems were compared for culture positivity and time to detection (TTD). Twenty-four mixed cultures with a yeast and a bacteria were tested. Bactec Mycosis vials could detect yeasts in all 24 cultures. The aerobic vials from both Bactec and BacT/Alert could detect both yeasts and bacteria in 22/24 (91.66?%) cultures. Bactec Plus Anaerobic/F and BacT/Alert FN vials could detect both microorganisms in 19/24 (79.16?%) and 4/24 (16.67?%) vials, respectively. Seven polymicrobial sepsis models with C. albicans + C. glabrata were also tested. Mycosis vials could detect both yeasts in 7/7 mixed cultures. The aerobic vials from Bactec and BacT/Alert could detect both yeasts in 3/7 and 2/7 mixed cultures, respectively. Bactec Plus Aerobic/F had a shorter TTD compared to BacT/Alert FA and Bactec Plus Anaerobic/F vials (p?<?0.0001 and p?<?0.01, respectively). The present study shows that the Bactec and BacT/Alert systems have different characteristics in the detection of yeasts and bacteria with polymicrobial sepsis.     

Randomised controlled trial of intensive multifactorial treatment for cardiovascular risk in patients with screen-detected type 2 diabetes: 1-year data from the ADDITION Netherlands study

Background

A growing body of evidence suggests that earlier diagnosis and treatment of diabetes may be beneficial; however, definitive evidence is lacking.

Aim

To evaluate the effectiveness of an intensified multifactorial treatment on cardiovascular risk factors in patients with screen-detected type 2 diabetes.

Design of study

Randomised controlled trial.

Setting

Seventy-nine general practices in the southwestern region of the Netherlands.

Method

In this randomised trial, patients diagnosed with diabetes by screen-detection were assigned to intensified (n = 255) or routine treatment (n = 243), and followed over 1 year. Intensified treatment consisted of pharmacological treatment combined with lifestyle education to achieve haemoglobin A1c (HbA1c) <7.0%, blood pressure <135/85 mmHg, and cholesterol <5.0 mmol/l (4.5 mmol/l if cardiovascular disease was present). Health-related quality of life (HRQoL) was assessed using the Short Form (SF)-36. Analyses were performed using generalised estimating equations models.

Results

Changes in body mass index were 0.2 (routine care) versus −1.4 kg/m² (intensified treatment), P<0.001; systolic blood pressure −19 versus −33 mmHg, P<0.001; diastolic blood pressure −7 versus −12 mmHg, P<0.001; HbA1c −0.9% versus −1.1%, P = 0.03; cholesterol −0.5 versus −1.2 mmol/l, P<0.001; high-density lipoprotein cholesterol 0.1 versus 0.1 mmol/l, P = 0.26; low-density lipoprotein cholesterol −0.5 versus −1.0 mmol/l, P<0.001; triglycerides −0.3 versus −0.4 mmol/l, P = 0.71. No difference in HRQoL between the two groups was reported.

Conclusion

Intensified multifactorial treatment of patients with screen-detected diabetes in general practice reduces cardiovascular risk factor levels significantly without worsening HRQoL.     

Direct comparison of the BACTEC 9240 and BacT/ALERT 3D automated blood culture systems for candida growth detection

A direct comparison of two automated blood culture systems was conducted to compare their ability to detect Candida growth. The systems evaluated were the BACTEC 9240 (Bactec) and BacT/ALERT 3D (BacT). The aerobic, anaerobic, and mycology media for each system were evaluated: Bactec Plus Aerobic/F, Plus Anaerobic/F, and Myco/F Lytic bottles, respectively, and BacT FA, SN, and MB bottles, respectively. Each blood culture bottle was inoculated with fresh blood from healthy donors. Fifty isolates of Candida spp. were used. The six different blood culture bottles were each inoculated with 1000 yeasts per bottle and then incubated in the corresponding automated system. The BacT detected growth of 90% (135 of 150) of Candida pathogens, while Bactec detected 66% (100 of 150). Growth was detected in all BacT and Bactec mycology bottles, all BacT aerobic bottles, and by terminal subculture of all bottles. Sixty-five of 300 (22%) bottles had no growth detected; 50 from the Bactec (5 aerobic and 45 anaerobic) and 15 from the BacT (all anaerobic). Terminal subculture of "negative" bottles demonstrated viable yeast growth from all 65 bottles, representing 65 false-negatives. The mean time to growth detection in the BacT system was 25.62 h while the Bactec was 27.30 h (P < 0.01). Both automated blood culture systems detected all episodes of simulated candidemia when specialized mycology media were used. However, when only standard aerobic and anaerobic media were used, the BacT performed better than the Bactec in overall growth detection, time to growth detection, and number of false-negatives.     

Impact of Blood Volume,Tube Shaking,and Incubation Time on Reproducibility of QuantiFERON-TB Gold In-Tube Assay

Gamma interferon (IFN-γ) release assays (IGRAs) are functional assays used serially to measure the efficacy of novel tuberculosis (TB) vaccines and to screen health care workers for latent tuberculosis infection (LTBI). However, studies have shown nonreproducible IGRA results. In this study, we investigated the effects of blood volume (0.8, 1.0, and 1.2 ml), tube shaking (gentle versus vigorous), and incubation duration (16, 20, and 24 h) on the reproducibility of QuantiFERON-TB Gold In-Tube (QFT-GIT) results for 50 subjects (33 uninfected and 17 infected). The median IFN-γ TB response (TB antigen [Ag] minus nil value) was significantly higher with 0.8 ml blood (1.04 IU/ml) than with 1.0 ml (0.85 IU/ml; P = 0.002) or 1.2 ml (0.49 IU/ml; P < 0.001) for subjects with LTBI. Compared with 0.8 ml (11.8%), there were larger proportions of false-negative results with 1.0 ml (29.4%; P = 0.2) and 1.2 ml (41.2%; P = 0.05) of blood for infected subjects. Blood volume did not significantly change the proportions of positive results in uninfected controls. Compared with gentle shaking, vigorous shaking increased the median IFN-γ response in nil (0.04 versus 0.06 IU/ml; P < 0.001) and TB Ag (0.12 versus 0.24 IU/ml; P = 0.004) tubes and increased TB responses (TB Ag_vigorous minus nil_gentle) (0.02 versus 0.08 IU/ml; P = 0.004). The duration of incubation did not have a significant impact on the proportion of positive results in uninfected or infected subjects. This study identified blood volume and tube shaking as novel preanalytical sources of variability which require further standardization in order to improve the quality and reproducibility of QFT-GIT results.     

Controlled Comparative Evaluation of BacT/Alert FAN and ESP 80A Aerobic Media as Means for Detecting Bacteremia and Fungemia

During a one-year period, a total of 6,305 blood cultures were processed in a tertiary-care teaching hospital; 6 to 12 ml of blood was inoculated into both a BacT/Alert Fan aerobic bottle and an ESP 80A aerobic bottle. The FAN aerobic bottle contains an antimicrobial-absorbing material; the 80A aerobic bottle does not. Bottles were processed on their respective continuous-monitoring blood culture instruments for up to five days of incubation. Four hundred thirty-three cultures (6.9%) representing 301 septic episodes in 235 different patients yielded 490 bacteria or yeasts thought to be clinically significant. Two hundred seventy-five of the 433 presumed clinically significant positive cultures (63.5%) representing 195 septic episodes and yielding 301 isolates were positive in both FAN and 80A bottles. One hundred nine significant positive cultures (25.2%) (i.e., cultures positive with an organism judged to be of probable clinical significance) from 70 septic episodes yielded 126 isolates only in FAN bottles. Conversely, the 80A bottle was exclusively positive in 49 instances (11.3%), representing 36 septic episodes and yielding 63 isolates. The higher rates of significant positive blood cultures, numbers of septic episodes documented, and numbers of isolates recovered in FAN bottles versus 80A bottles were all statistically significant (P < 0.05). Enhanced rates of detection of presumed clinically significant isolates in FAN bottles were largely accounted for by Staphylococcus aureus, members of the Enterobacteriaceae, and non-Pseudomonas aeruginosa miscellaneous gram-negative bacilli from patients receiving antimicrobial therapy at the time blood cultures were obtained. Enhanced recovery of one organism group, the β-hemolytic streptococci, occurred in 80A. With one exception, detection times were essentially equivalent in the two systems. The single exception pertained to streptococci and enterococci, which were recovered significantly faster in 80A bottles. Three hundred thirty-eight of the 6,305 blood cultures evaluated in this study (5.4%) were judged likely to be contaminated. The percentages of probable contaminated cultures were as follows: 26.6% FAN and 80A; 42.3% FAN only; 31.1% 80A only (P < 0.05). Finally, the instrument false-positive rates for the two systems were 0.7% with FAN and 3.0% with 80A (P < 0.05). We conclude that while contamination rates were slightly higher with FAN than with 80A, use of FAN aerobic bottles in conjunction with the BacT/Alert system will yield significantly higher numbers of clinically significant blood culture isolates than 80A bottles and the ESP system. Furthermore, this enhanced detection is most conspicuous in patients receiving antimicrobial therapy at the time blood cultures are performed, probably due to the presence of an antimicrobial-absorbing material in FAN aerobic bottles.     

Nationwide Study of Candidemia,Antifungal Use,and Antifungal Drug Resistance in Iceland, 2000 to 2011

Candidemia is often a life-threatening infection, with highly variable incidence among countries. We conducted a nationwide study of candidemia in Iceland from 2000 to 2011, in order to determine recent trends in incidence rates, fungal species distribution, antifungal susceptibility patterns, and concurrent antifungal consumption. A total of 208 infection episodes in 199 patients were identified. The average incidence during the 12 years was 5.7 cases/100,000 population/year, which was significantly higher than that from 1990 to 1999 (4.3/100,000/year; P = 0.02). A significant reduction in the use of blood cultures was noted in the last 3 years of the study, coinciding with the economic crisis in the country (P < 0.001). Age-specific incidence rates were highest among patients at the extremes of age, 20.7/100,000 for <1 year of age and 18.1/100,000 for >60 years, and varied by gender. Age-specific incidence among males >80 years old was 28.6/100,000/year, and it was 8.3/100,000/year for females in this age group (P = 0.028). The 30-day survival rate among adult patients remained unchanged compared to that from 1990 to 1999 (70.4% versus 69.5%, P = 0.97). Candida albicans was the predominant species (56%), followed by C. glabrata (16%) and C. tropicalis (13%). The species distribution remained stable compared to that from previous decades. Fluconazole use increased 2.4-fold from 2000 to 2011, with no increase in resistance. In summary, the incidence of candidemia in Iceland has continued to increase but may have reached a steady state, and no increase in antifungal drug resistance has been noted. Decreased use of blood cultures toward the end of the study may have influenced detection rates.     

Prevalence of virulent Candida spp. in complicated urinary tract infection of nephrolithiatic patients from surgical units of tertiary care hospitals Islamabad

Candida species are the commensal organisms of human mucosa and opportunistically cause the diseases in susceptible persons. This study aimed to determine the prevalence and virulence of different Candida spp. among nephrolithiatic patients and their association with complicated UTI (cUTI). A total of 164 urine samples were collected from surgical units of two tertiary care hospitals (Poly Clinic and Pakistan Institute of Medical Sciences Hospital, Islamabad). From 74 kidney stone patients, 77 isolates of Candida spp. were confirmed through standard microbiological and molecular characterization. C. albicans was the predominant species with 51 isolates (66.2%) followed by 26 (33.8%) of C. non-albicans. The nephrolithiatic patients suffering from cUTI were more prone to be infected with Candida (P = 0.047). Among all isolates, 83% (64) of the Candida isolates were biofilm formers, 80% (60) showed the esterase production and 64.9% (50) showed phospholipase production. Candida isolates positive for various virulence factors were more prevalently isolated from both catheterized and recurrent UTI patients. Among Candida spp., 16.9% (13) isolates showed resistance to fluconazole and 19.5% (15) against voriconazole and 11 isolates were resistant for both tested antifungals. Candida isolated from cUTI cases showed comparatively enhanced virulence attributes and antifungal resistance, suggesting that these factors might have role in development of cUTI in nephrolithiatic patients. Hence, this work highlights the high prevalence of both C. albicans and non albicans spp. in nephrolithiatic patients. So, there is need to administer evidence based antifungal therapy rather than empirical therapy to reduce the cUTI in nephrolithiatic patients.     

Role of the Mycobiome in Human Acute Graft-versus-Host Disease

A role for gut bacteria in the pathogenesis of graft-versus-host disease (GVHD) has been firmly established; however, the role of Candida spp, which form part of the mycobiome, remains unknown. In a homogenous group of patients who underwent allogeneic stem cell transplantation (SCT), we found a significant impact of Candida colonization on the occurrence of acute GVHD. Patients colonized with Candida spp developed significantly more grade II-IV acute GVHD compared with noncolonized patients (50% vs 32%; P = .03), as well as more gastrointestinal (GI)-GVHD (33% vs 19%; P = .05). Colonization with Candida spp was more frequent in patients bearing the loss-of-function polymorphism Y238X, which results in dectin-1 dysfunction, compared with patients with the wild-type allele (73% vs 31%; P = .002). There was no direct effect of dectin-1 dysfunction on acute GVHD, although it did influence the occurrence of GVHD indirectly through Candida colonization. The exact mechanism of GVHD induction by Candida spp colonization of the mucosa is unknown, but the link might prove to be the induction of Th 17/IL-23 responses through activation of pattern recognition receptors by fungal motifs, including β-d-glucan and mannans. These data indicate a role for the mycobiome in the pathogenesis of GVHD and suggest that altering the mycobiome by antifungal drugs can help ameliorate GI-GVHD. In addition, given that the genetic constitution of patients affects susceptibility to both Candida colonization and GVHD, whether identifying gene polymorphisms will facilitate personalized treatment of SCT recipients remains to be determined.