Evaluation of blood culture bottles seeded with X-V factors for the detection of Neisseria meningitidis and Haemophilus influenzae.

The purpose of this in vitro study was to determine the ability of seeded and not-seeded commercial pediatric blood culture bottles to support the growth of the most frequently responsible microorganisms for bacterial meningitides (Neisseria meningitidis, and Haemophilus influenzae). Tests have been carried out with an automated colorimetric pediatric blood culture system, BacTAlert, Organon Teknika. Bottles were inoculated with X-V factors and serial dilutions of the each bacterium in six times (10(1)-10(6) colony forming unit [CFU]/ml). The bottles, which were supplemented with X-V factors, proved to be effective and time to detection (TTD) was shorter than the un-seeded bottles (p0.05). Time difference between seeded and not-seeded bottles was getting greater at high dilutions of both bacteria. We consider that in presence of a few bacteria, the seeding of bottles with X-V factors is very critical obtaining N. meningitidis, and H. influenzae as the causative agents of meningitidis. The recovery rate of the microorganisms, which were isolated from cerebrospinal fluid by using the X-V factor-seeded blood culture bottles, is therefore higher than with the conventional culture methods.

Different mechanisms of Ca2(+)-handling following nicotinic acetylcholine receptor stimulation, P2U-purinoceptor stimulation and K(+)-induced depolarization in C2C12 myotubes.

1. The increase in intracellular CA2+ on nicotinic acetylcholine receptor (nAChR) stimulation, P2U-purinoceptor stimulation and K(+)-induced depolarization was investigated in mouse C2C12 myotubes by use of fura-2 fluorescence to characterize the intracellular organisation of Ca2+ releasing stores and Ca(2+)-entry process. 2. Stimulation of nAChRs with carbachol induced a rapid rise in internal Ca2+ (EC50 = 0.85 +/- 0.09 microM), followed by a sustained phase. The Ca2+ response evoked by carbachol (10 microM) was completely blocked by the nAChR antagonist, pancuronium (3 microM), but was not affected by the muscarinic antagonist, atropine (3 microM), or under conditions when Ca2+ entry was blocked by La3+ (50 microM) or diltiazem (10 microM). Addition of pancuronium (3 microM) during the sustained phase of the carbachol-evoked response did not affect this phase. 3. Stimulation of P2U purinoceptors with ATP (1 mM) induced a somewhat higher biphasic Ca2+ response (EC50 of the rapid phase: 8.72 +/- 0.08 microM) than with carbachol. Pretreatment with La3+ abolished the sustained phase of the ATP-induced Ca2+ response, while the response was unaffected by diltiazem or pancuronium. 4. Stimulation of the cells with high K+ (60 mM), producing the same depolarization as with carbachol (10 microM), induced a rapid monophasic Ca2+ response, insensitive to diltiazem, pancuronium or La3+. 5. Under Ca(2+)-free conditions, the sustained phase of the carbachol- and ATP-evoked responses were abolished. Pre-emptying of depolarization-sensitive stores by high K+ under Ca(2+)-free conditions did not affect the carbachol- or ATP-evoked Ca2+ mobilization and vice versa. Preincubation of the cells with ATP in the absence of extracellular Ca2+ decreased the amplitude of the subsequent carbachol-induced Ca2+ response to 11%, while in the reverse procedure the ATP-induced response was decreased to 65%. Ca2+ mobilization evoked by simultaneous addition of optimal concentrations of carbachol and ATP was increased compared to levels obtained with either agonist. 6. Preincubation with high K+ under normal conditions abolished the sustained phase of the ATP-evoked Ca2+ response. The carbachol response consisted only of the sustained phase in the presence of high K+. 7. The carbachol-induced Ca2+ response was completely abolished under low Na+/Ca(2+)-free conditions, while under low Na+ conditions only a sustained Ca2+ response was observed. The ATP- and K(+)-induced responses were changed compared to Ca(2+)-free conditions. 8. ATP (300 microM) induced the formation of Ins(1,4,5)P3 under Ca(2+)-free conditions with a comparable time course to that found for the rise in internal Ca2+. In contrast to ATP, carbachol (10 microM) did not affect Ins(1,4,5)P3 levels under Ca(2+)-free conditions. 9. It is concluded that the Ca2+ release from discrete stores of C2C12 myotubes is induced by stimulation of nAChRs, P2U-purinoceptors and by high K+. Only the P2U-purinoceptor and nAChR activated stores show considerable overlap in releasable Ca2+. Sustained Ca(2+)-entry is activated by stimulation of nAChRs and P2U-purinoceptors via separate ion-channels, which are different from the skeletal muscle nAChR-coupled cation-channel.

Evaluation of the automated Bact-Alert system for pediatric blood culturing.

The Organon Teknika BacT/Alert (Organon Teknika, Durham, NC), using the Pedi-BacT 20 mL aerobic bottle (BPBCS) was compared to the Wampole Isolator (WI) 1.5 Microbial tube (Wampole Laboratories, Cranbury, NJ), for detection and recovery of pediatric pathogens. The BPBCS continuously monitors culture bottles for changes in CO2 concentrations, while WI cultures are examined twice daily for appearance of colonial growth on agar media. Of 5,175 paired blood cultures, 383 pathogens were recovered from 606 positive cultures. There were 272 pathogens recovered by both systems, 64 from BPBCS only, and 47 from WI only. Overall recovery rates were 88% for BPBCS and 83% for WI. There was no significant difference between the two systems in detection or times to positivity of staphylococci, Enterobacteriaceae, or pseudomonads. Trends toward better recovery of streptococci (20 vs. 10) and fastidious microaerophiles (3 vs. 0) were found with BPBCS, whereas more slowly growing pathogens (Rochalimaea henselae [1], Mycobacterium avium-intracellulare [1]) were recovered by WI only, but because of their lower frequency did not achieve statistical significance. Detection of Haemophilus influenzae (14.9 hours in WI vs. 45.4 hours in BPBCS) was faster with WI. False positive plus contaminant cultures were detected in 5.9% BPBCS versus 1.5% WI. BPBCS offers detection of bacteremia at a rate comparable to WI with advantages of automation.

Use of simulated blood cultures for time to detection comparison between BacT/ALERT and BACTEC 9240 blood culture systems.

In order to avoid the influence of pre-analytical steps, the following study was performed by using sterile blood spiked with defined loads of microorganisms as inoculum. Time-to-Detection (TTD) was evaluated for the most frequently encountered bacteria and fungi in septicemia, comparing two commercially available blood culture systems, BACTEC 9240 (Becton Dickinson) and BacT/ALERT (Organon Teknika). A specific medium, Bactec Mycosis IC/F (Becton Dickinson), was compared with the Bactec Plus Aerobic (Becton Dickinson) and FAN Aerobic (Organon Teknika) media for recovery of fungi in general and in case of mixed bacterial/fungal septicemia. The results show that the BACTEC system detects nearly all enrolled microorganisms significantly faster than the BacT/ALERT; the anaerobic vial contributes to the detection of anaerobes and facultative anaerobes and, in the case of BACTEC, shortens TTD; the Bactec Mycosis IC/F bottle shortens TTD of fungi.

Evaluation of the MB/BacT system for recovery of mycobacteria from clinical specimens in comparison to Lowenstein-Jensen medium.

OBJECTIVE: To evaluate the performance of the MB/BacT system (Organon Teknika) in comparison to Lowenstein-Jensen (LJ) solid medium for recovery of mycobacteria from clinical specimens. METHODS: Two thousand three hundred and ten specimens (1626 respiratory, 593 urine, 60 body fluids, five tissue and 26 others) were inoculated in MB/BacT (0.5 mL) and on two LJ slants (0.25 mL each). N-acetyl-l-cysteine-NaOH (final concentration 2%) was used for decontamination. RESULTS: Two hundred and fifty-one (10.8%) mycobacterial isolates [190 Mycobacterium tuberculosis complex (MTBC) and 61 non-tuberculous mycobacteria (NTM)] were detected. Of these 251 isolates, 234 (93.2%; 181 MTBC and 53 NTM) were detected in MB/BacT and 169 (67.3%; 154 MTBC and 15 NTM) on LJ. The mean (median) times to detection of MTBC by MB/BacT and LJ were 13.8 (13) and 22.1 (20) days, respectively, while overall contamination rates were 7.7% and 8.1%, respectively. CONCLUSIONS: Sensitivity and time to detection were significantly better with MB/BacT than with solid LJ medium.

Screening of Salmonella in naturally contaminated feeds with rapid methods.

Five commercially available screening methods, the Oxoid MSRV, Merck SALMOSYST-RAMBACH AGAR combination, Organon Teknika SALMONELLA-TEKTM, Dynal DYNABEADS ANTI-SALMONELLA and Foss Electric EIAFOSS, were compared to the conventional culture procedure for the detection of Salmonella in naturally contaminated feed samples. A total of 217 feed samples from animal as well as from vegetable origin were examined. Twenty one samples were found to be positive for Salmonella by all methods combined. The conventional culture method detected 17 (81,0%), MSRV 19 (90,5%), SALMOSYST-RAMBACH 8 (38,1%), SALMONELLA-TEK 19 (90,5%), DYNABEADS ANTI-SALMONELLA 7 (33,3%) and EIAFOSS 21 (100%) of the 21 total Salmonella contaminated samples.

Identification of Listeria monocytogenes from unpasteurized apple juice using rapid test kits.

A microbiological survey of 50 retail juices was conducted in the fall of 1996. These juices were analyzed for Listeria monocytogenes, Escherichia coli O157:H7, Salmonella, coliforms, fecal coliforms, and pH. Two unpasteurized juices were positive for L. monocytogenes: an apple juice and an apple raspberry blend with a pH of 3.78 and 3.75, respectively. Three L. monocytogenes isolates were characterized. The colonies were typical for Listeria sp. on Oxford and lithium chloride-phenylethanol-moxalactam agars and were beta-hemolytic on sheep blood agar. The isolates required 5 days of incubation at 35 degrees C to produce a positive rhamnose reaction in a phenol red carbohydrate broth. This slow rhamnose utilization resulted in these isolates not being identified using the Micro-ID test strip (Organon Technika). However, the isolates were positive for L. monocytogenes using the API Listeria strip (BioMerieux) and a multiplex polymerase chain reaction for detection of the hemolysis (hyla) and invasion-associated protein (iap) genes.

Use of simulated blood cultures for antibiotic effect on time to detection of the two blood culture systems BacT/ALERT and BACTEC 9240.

To avoid the influence of pre-analytical steps, this study was performed using sterile blood spiked with defined loads of microorganisms as inoculum. Time-to-Detection (TTD) was evaluated for the most frequently encountered bacteria comparing two commercially available blood culture systems, BD BACTEC 9240 (Becton Dickinson) and BacT/ALERT (Organon Teknika). The effect of the most widely used antibiotics on TTD was evaluated on both systems. TTD was measured with antibiotics at their trough and at increasing concentrations. The results show that the BACTEC PLUS system recovers more pathogens with shorter time to detection than the BacT/ALERT FAN system when beta-lactam antibiotics (Ampicillin, Cefotaxime) are present at their respective trough concentration corresponding to parenteral therapy. The two systems seem to be equally efficient when Gentamicin, Ciprofloxacin and Trimethoprim/sulfamethoxazole are used; in the case of Vancomycin, BACTEC seems more effective than BacT/ALERT.

[Blood culture update]. / Actualités sur l'hémoculture.

BACKGROUND: Blood culture is one of the most important bacteriological examinations with important clinical and therapeutic consequences. Blood cultures should be ordered in all patients with signs suggesting septicemia, endocarditis or severe infection (pneumococcal pneumonia, bacterial meningitis with bloodstream dissemination). Blood culture methods have evolved considerably over the last twenty years. After using manual methods for many years, read by non-standardized visual methods, the development of media with defined compositions and supplemented to allow growth of bacteria difficult to culture has been associated with the development of automatic blood culture devices. AUTOMATIC DEVICES: These devices have undergone rapid improvement. Semi-automatic devices (Bactec NR-660) were rapidly followed by completely automatic techniques, including four devices currently available: since 1989 Bio-Argos (Rio-Rad) and Bact/Alert (Organon-Teknika) and in 1993, Bactec 9240 (Becton-Dickinson) and Vital (BioMérieux). All these devices allow automatic detection of CO2 produced during bacterial growth. Automatic reading systems provide continuous output avoiding the need for invasive methods and thus the risk of contamination in addition to saving time. Potential application to achieve quantitative blood cultures for intensive care units is in the development stage. CONSEQUENCES: The reliability of these devices is well recognized and their contribution to severe bacterial infection is undeniable. There are certain limitations however related to material cost and the non-identification of the pathogen involved. Molecular biology techniques open new perspectives in this field. The evolution of techniques, definitions, and pathogenic approach to septicemia must be revisited as new infectious situations have been identified at the same time as new investigation tools resulting from considerable technological progress. New methods of blood culture have largely contributed to this progress.

[Comparative evaluation of BACTEC MGIT 960 system with MB/BacT and egg-based media for recovery of mycobacteria].

Fully automated, nonradiometric mycobacteria culture systems, BACTEC MGIT 960 (Becton Dickinson Microbiology Systems, Sparks, Md, U.S.A.) and MB/BacT (Organon Teknika, Durham, NC, U.S.A.) were evaluated in comparison with three different eggbased media (3% Ogawa, Ogawa K, and Vite) for the ability to detect mycobacteria in clinical sputum specimens. Sputum specimens were processed by semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) for the automated systems, and by cetylpyridium chloride-succinic acid-NaCl for the egg-based media. A total of 954 sputum specimens were processed, and the recovery of mycobacteria by the BACTEC MGIT 960 was performed in a commercial laboratory. Overall, the frequency of breakthrough contamination was <1% for the three egg-based media, ranging from 0. 42% to 0.63%. Whereas, the frequency of false positives due to breakthrough contamination was 1.89% for MB/BacT and 20.1% for BACTEC MGIT 960. A total of 237 isolates of Mycobacterium tuberculosis complex and 167 isolates of nontuberculous mycobacteria (NTM) were recovered. The highest recovery ratio was obtained by MB/BacT (95.8%), followed by the egg-based media, Vite (74.3%), Ogawa K (65.8%), and 3% Ogawa (58.9%). The recovery ratio by BACTEC MGIT 960 was the lowest, and estimated at 43.1%, mainly due to a high frequency of breakthrough contamination. However, even after omission of these false positives reported by BACTEC MGIT 960, the recovery ratio by this system was comparable to that of 3% Ogawa media. The time to detection of 50% of positive cultures of M. tuberculosis complex by BACTEC MGIT 960 and MB/BacT was 20 days and 17 days, respectively.>From these results, it may be concluded that MB/BacT is superior to BACTEC MGIT 960 and egg-based media for the recovery of mycobacteria from sputum specimens. Furthermore, based on the outcome of this study, we think that considerable improvements are necessary for the clinical application of BACTEC MGIT 960. These improvements should particularly be focused on reducing the false positive ratio caused by contamination, and culture media, which effectively support mycobacterial growth.

[Application of DNA-DNA hybridization, DDH MYCOBACTERIA 'Kyokuto' to species-identification of mycobacteria grown in Middlebrook 7H9 broth].

A colorimetric DNA-DNA hybridization for the genetic identification of mycobacteria, DDH MYCOBACTERIA 'KYOKUTO' (Kyokuto Pharmaceuticals, Tokyo) was evaluated for the clinical isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT (Organon Teknika, Durham, NC, U.S.A.). When the MB/BacT gave the positive interpretation, 10 ml of Middlebrook 7H9 broth was collected from the bottle. After centrifugation at 3,000 rpm for 10 min, two drops of acetone were added to the pellet, then let it stand for one hour at the room temperature. The air-dried pellet was resuspended in a small volume of distilled water, and proceeded to the identification described. Of 136 clinical isolates of mycobacteria, comprising of 76 M. tuberculosis complex and 60 nontuberculous mycobacteria (NTM), ninety-five (70%) were correctly identified when compared to the reference identification. Thirty (22%) isolates resulted in the unidentified due to negative reaction throughout the test wells, and the remaining 11 (8%) were also unidentified due to low likelihoods. According to the package insert, the DDH MYCOBACTERIA may not be applicable to the isolates grown in Middlebrook broth. However, our test procedure using acetone prior to the extraction of bacterial DNA enables us to directly identify the isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT.

[Evaluation of a fully automated mycobacteria culture system, MB/BacT using a newly developed digestion-decontamination procedure, semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) method].

A fully automated non-radiometric mycobacteria culture system, MB/BacT (Organon Teknika, Durham, NC, U.S.A.), was recently introduced in Japan and evaluated for its ability to detect mycobacteria in clinical sputum specimens. A previous study yielded nearly a 40% contamination ratio from sputa treated with the standard N-acetyl-L-cysteine (NALC)-NaOH method. This study employed a mucolytic agent (semi-alkaline protease; SAP) in which the sputa were processed twice for digestion followed by decontamination at twice the standard volume of NALC-NaOH. The concentrated sediments were resuspended in phosphate buffer (0.067 M, pH 6.8), and inoculated into the MB/BacT Process Bottles supplemented with antibiotics. The bottles were incubated at 37 degrees C and monitored for up to fifty-six days. Recovery of mycobacteria was compared in three different egg-based Ogawa media in addition to a non-selective Middlebrook 7H10 agar. A total of 1, 124 clinical sputum specimens have been evaluated. Of these, 464 were positive for growth of mycobacteria, of which 447 (96.3%) were positive by the MB/BacT. False-positive alarms due to break through contamination, mainly by Pseudomonas aeruginosa and Candida spp., were observed in twenty-one specimens (1.9%). The three Ogawa media could detect only 283 (60.5%) to 353 (75.4%) positives, and Middlebrook 7H10 agar only 424 (90.6%) positives. The time to detect positive cultures of Mycobacterium tuberculosis complex by the MB/BacT ranged from 2.2 days to 52.3 days, and 50% of positive cultures were detected within 16.7 days of incubation. It can be concluded that the combination of SAP-NALC-NaOH digestion-decontamination procedure and the MB/BacT is particularly useful for the isolation of mycobacteria and has a faster time to detect than conventional methods. MB/BacT is a suitable alternative method for the detection of mycobacteria in Japan, where the radiometric Bactec System is not available.

[Comparative evaluation of two different formulae of Middlebrook 7H9 broth in a fully automated mycobacteria culture system, MB/BacT; the effect of Tween 80].

Two different formulae of Middlebrook 7H9 broth, one containing Tween 80 [Tween (+) broth] and the other containing vancomycin but not Tween 80 [Tween (-) broth], were evaluated in parallel for a fully automated mycobacteria culture system, MB/BacT(Organon Teknika, Durham, NC, U.S.A.). A total of 586 clinical sputum specimens were digested and decontaminated by the semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH). Each part of sample treated was inoculated into the MB/BacT Process Bottle containing the respective Middlebrook 7H9 broth. Culture bottles were incubated in the MB/BacT at 37 degrees C for up to 56 days. Of 586 samples, 110 isolates of Mycobacterium tuberculosis complex and 77 of nontuberculous mycobacteria (NTM) were isolated. The occurrence of false alarm due to breakthrough contamination was 3.2 in Tween (+) broth and 2.9% in Tween (-) broth. Also, the positivities of mycobacteria by the respective culture media were comparable. However, Tween (-) broth could detect positive cultures for mycobacteria, particularly for M. tuberculosis complex at the earlier incubation cycle when compared to Tween (+) broth. The time to detect 50% positive cultures for M. tuberculosis complex was 20.5 days for Tween (-) broth and 34.3 days for Tween (+) broth, respectively. With the results, it was concluded that; Tween (+) broth produced homogeneous mycobacterial growth in culture media, and thus, it was easy to prepare the inoculum directly adjusted to McFarland turbidity to the susceptibility test. However, the present formula of Middlebrook 7H9 broth supplemented with Tween 80 was not enough suitable for the rapid detection of positive cultures and needs some revisions to improve.

[Effects of medicinal insect, Catharsius molossus on biosynthesis of polysaccharides from Ganoderma lucidum in submerged culture].

We studied the effects of several medicinal insects on biosynthesis of polysaccharides from Ganoderma lucidum in submerged culture. The results showed that the medicinal insect, Catharsius molossus at 5 g/L significantly promoted the biosynthesis of intracellular polysaccharides (IPS) and extracellular polysaccharides (EPS) of G. lucidum, and compared with control, IPS and EPS yields markedly enhanced from (1.93 +/- 0.09) g/L to (2.41 +/- 0.12) g/L and (520.3 +/- 20.2) mg/L to (608.9 +/- 20.2) mg/L, respectively (P < 0.05). Both IPS and EPS consisted of five kinds of components, and IPS-1 and EPS-1 were the major components of IPS and EPS, respectively. Further separation studies showed that IPS-1 was made up of three single compounds, while EPS-1 was made up of two single compounds. There were no new components in both IPS and EPS obtained from G lucidum in submerged culture by the addition of the insect, C. molossus, suggesting the biosynthetic pathways of the major components of IPS and EPS had not been changed.

[Effects of medicinal insect Catharsius molossus on submerged fermentation and in vivo anti-hepatoma activity of Ganoderma lucidum].

We studied the effects of Catharsius molossus (a Chinese medicinal insect) on the cell growth, fermentation kinetics of key bioactive substances and anti-cancer activity of Ganoderma lucidum in submerged fermentation. The results showed that C. molossus at all the tested concentrations had no stimulatory effect on the cell growth. However, addition of C. molossus at 5 g/L lead to significant effects on the fermentation kinetics of polysaccharides and triterpenoids of G lucidum, and at 7th day in fermentation process, the yields of polysaccharides and triterpenoids reached 2.81 g/L and 539.0 mg/L, respectively, while they were 2.25 g/L and 428.2 mg/L in control. In vivo anti-cancer studies showed that the inhibitory rates of control fermented G lucidum (CFG) and a combination of water extract from C. molossus and CFG on the developed tumor (Heps) in mice were 41.61% and 42.24%, respectively. Moreover, the inhibitory rate of the G lucidum fermented with C. molossus (GFC) reached 57.21%, which was enhanced 37.49%, compared to the inhibitory rate of the control fermented G lucidum. These results suggest that supplementation of C. molossus in submerged fermentation of G lucidum lead to a significant enhancement of the anti-cancer activity of cultured G lucidum.

[Effect of musk soluble components on the growth, the differentiation and the transfection efficiency of rat neural stem cells (NSC) in vitro].

AIM: To investigate the effect of musk soluble components on the growth, the differentiation and the transfection efficiency of rat neural stem cell (NSC) in vitro. METHODS: The growth and the differentiation of rat NSC were observed when musk soluble components were added into the culture medium of NSC. Meanwhile, the pEGFP-C1, which expressed the enhanced GFP protein, was transfected into the NSC by method of electro- transfection. RESULTS: When NSC was treated with musk soluble components, the neurites were outgrowth around NSC and attached to the plate, and the neural spheres were disassociated. The glia-like cells appeared at the concentration of 0.3 per thousand. When the concentration of musk soluble components was lower than 3 per thousand, the transformative cells could recover. Furthermore, the efficiency of transfection pEGFP into NSC was remarkably increased after the treatment with musk. CONCLUSION: After the treatment of NSC with musk soluble components, the neural spheres were disassociated, and then attached to the plate. Musk soluble components could induce NSC differentiation into glia-like cells and improve the transfection efficiency of pEGFP-C1 in vitro.

Diagnosis of candidemia by polymerase chain reaction and blood culture: prospective study in a high-risk population and identification of variables associated with development of candidemia.

The diagnosis of candidemia is important for prompt initiation of antifungal therapy. Two hundred twenty-five patients at high risk for candidemia who had blood cultures drawn and were hospitalized for more than 15 days were followed-up prospectively over a 2-year period. Polymerase chain reaction (PCR) and whole-blood cultures monitored by the automated BactAlert system (Organon Teknika, Durham, NC, USA) were used to detect candidemia in all patients hospitalized in high-risk areas for more than 15 days. DNA was extracted and amplified using ITS5 and ITS4 base pair primers, and the PCR products were sequenced for identification of Candida spp. A blood culture positive for Candida was considered the gold standard for diagnosis of candidemia. Variables associated with the development of candidemia diagnosed by positive blood culture were also evaluated in the patients. The overall mortality rate was 26.1%. Mortality in candidemic patients was 41.9% and in noncandidemic patients 22.5% (p = 0.009). PCR sensitivity and specificity were 72.1 and 91.2%, respectively. Positive and negative predictive values were 65.9 and 93.2%, respectively. The logistic regression of the multivariate analysis showed that parenteral nutrition (p < 0.0001), fever (p = 0.01), neutropenia (p = 0.04), and an indwelling urinary catheter (p = 0.02) were significant variables associated with the development of candidemia. The PCR technique in conjunction with DNA sequencing was a helpful tool in the diagnosis of candidemia.

Detection of Campylobacter upsaliensis from a blood culture by using the BacT/Alert system.

Campylobacter upsaliensis was isolated from the blood of a 60-year-old female with hairy cell leukemia. This spiral-shaped organism was detected in the aerobic BacT/Alert bottle (Organon Teknika, Durham, N.C.) by acridine orange staining and was recovered only on chocolate agar in a microaerophilic atmosphere at 35 degrees C.

Assessment of routine use of an anaerobic bottle in a three-component, high-volume blood culture system.

The relative value of routine anaerobic blood culture for recovery of organisms and identification of episodes of bloodstream infection was assessed in a three-component, high-volume blood culture system which employs aerobic and anaerobic bottles of BacT/Alert (Organon-Teknika, Durham, N.C.) and aerobic cultures of Isolator (Wampole Laboratories, Cranbury, N.J.). The results of 5,595 blood culture sets from patients with suspected bloodstream infection were analyzed. Compared with either the aerobic BacT/Alert bottle or aerobic culture of Isolator, the BacT/Alert anaerobic bottle recovered significantly fewer isolates (242 versus 294, P < 0.05; 242 versus 298, P < 0.05) but did not detect significantly fewer episodes of bloodstream infection (141 versus 157, P > 0.05; 141 versus 147, P > 0.05). The BacT/Alert anaerobic bottle recovered significantly more isolates of obligately anaerobic bacteria (16 versus 4, P < 0.05; 16 versus 0, P < 0.05) and detected significantly more episodes of bloodstream infection caused by obligately anaerobic bacteria (10 versus 3, P < 0.05; 10 versus 0, P < 0.05) than either the aerobic bottle of BacT/Alert or the aerobic culture of Isolator. The combination of the BacT/Alert anaerobic bottle and the aerobic culture of Isolator recovered as may isolates (374 versus 377) and detected as many episodes of bloodstream infection (194 versus 191) as the combination of the aerobic bottle of BacT/Alert and the aerobic culture of Isolator, and both of these combinations identified at least 8% more isolates and detected at least 3% more bloodstream infections than the combination of the BacT/Alert aerobic and anaerobic bottles. Further analysis of the data revealed that the utility of the BacT/Alert anaerobic bottle, especially when combined with the aerobic culture of Isolator, resulted from not only enhanced recovery of obligately anaerobic bacteria but also effective recovery of facultatively anaerobic bacteria. These results demonstrate the utility of the anaerobic BacT/Alert bottle for detecting bloodstream infection caused by either facultatively anaerobic bacteria or obligately anaerobic bacteria and support the routine inclusion of anaerobic blood culture in the three-component blood culture system used in our hospital.