Growth and detection of filamentous fungi in the BacT/Alert blood culture system.

Little is known about the behaviour of filamentous fungi in most blood culture systems, despite their increasingly recognised role in infections of immunocompromised hosts. The ability of the BacT/Alert system (Organon Teknika, Durham, North Carolina, USA) to detect the growth of 19 such fungi was examined. Eleven species grew and were detected rapidly; two species did not grow. Six species grew slowly, and were generally only recovered with terminal subculture after prolonged incubation. The CO2 production graph for some of these fungi showed a slow but steady rise, insufficient to cause the apparatus to signal positive. These results show that the BacT/Alert system may miss some fungi, either because of no growth in the medium or undetected slow growth. The latter problem could be overcome by prolonged incubation and terminal subculture when fungal infection is considered likely. Alteration of the signalling mechanism might permit earlier detection of some slow growing fungi.

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.

Controlled comparison of original vented aerobic fan medium with new nonvented BacT/ALERT FA medium for culturing blood.

To evaluate the performance of BacT/ALERT FA (FA) medium, a new aerobic BacT/ALERT FAN (FAN) medium (Organon Teknika Corporation, Durham, N.C.) that does not require the added cost and inconvenience of a venting unit, we inoculated blood specimens from adult patients with suspected sepsis into an original FAN aerobic culture bottle and an FA bottle. Of 7,745 blood culture sets containing both bottles, 5,256 (68%) met the criteria for adequacy of filling. A total of 466 isolates judged to represent the causes of true infections were recovered from 276 patients; 271 isolates were recovered from both bottles, 82 were recovered from the FAN bottle only, and 113 were recovered from the FA bottle only (P < 0.05). More Burkholderia cepacia isolates (P < 0.01), Candida albicans isolates (P < 0.001), Cryptococcus neoformans isolates (P < 0.01), yeasts overall (P < 0.001), and total microorganisms (P < 0.05) were recovered from FA bottles. Of cultures found to be positive within the first 72 h of incubation, the mean times to detection were almost identical for FAN (20.4 h) and FA (20.7 h) bottles. Of 263 isolates that caused monomicrobic episodes of bloodstream infections, 180 were detected in both bottles, 32 were detected in FAN bottles only, and 51 were detected in FA bottles only (P < 0.05). Of 186 isolates considered to be contaminants, 63 were detected in both media, 64 were detected in FAN bottles only, and 59 were detected in FA bottles only (P was not significant). The number of false-positive results were comparable: 69 (1.3%) in FAN bottles and 56 (1.1%) in FA bottles. However, there were 14 isolates with false-negative results (6 yeasts, 6 nonfermenters, and 1 isolate each of Propionibacterium acnes and coagulase-negative staphylococci) in FAN bottles, whereas there were none in FA bottles. On the basis of these results, we conclude that the new nonvented FA bottle is superior to the original vented FAN medium for the recovery of B. cepacia and yeasts, especially C. albicans and C. neoformans, and is comparable to FAN medium for other microorganisms.

Clinical importance of increased sensitivity of BacT/Alert FAN aerobic and anaerobic blood culture bottles.

Two recent multicenter blood culture studies found that BacT/Alert FAN (FAN) bottles (Organon Teknika, Durham, N.C.) had increased yields in detecting bacteremia and fungemia compared with standard BacT/Alert (STD) bottles. Because the clinical importance of this increase in microbial recovery is unknown, we performed a retrospective analysis to determine the frequency with which FAN bottles were the sole means of detecting an episode of bacteremia. There were 1,047 positive blood cultures in which both study bottles were adequately filled and the organism isolated was judged to be the cause of sepsis: 240 (23%) were positive only in FAN bottles and 73 (7%) were positive only in STD bottles. Of a total of 664 episodes of bacteremia, 126 (19%) were identified only by FAN bottles and 43 (7%) were identified only by STD bottles (P < 0.0001). Episodes detected only by FAN bottles more often were recurrent events (23 of 126, or 18%) than episodes detected only by STD bottles (2 of 43, or 5%) (P < 0.05) and more commonly occurred in patients receiving theoretically effective antibiotic therapy (33 of 126 [26%] versus 4 of 43 [9%]) (P < 0.05). The medical records for patients with 127 of these episodes (92 FAN bottles only; 35 STD bottles only) were available for review. More than half of both FAN bottle-only (60 of 92, or 65%) and STD bottle-only (20 of 35, or 57%) episodes were judged to be clinically important. We conclude that FAN bottles improve the detection of bacteremia and that the majority of the additional episodes detected are clinically important. The benefits of the greater yield in specific patient populations must be balanced against the higher costs of FAN bottles.

BacT/Alert: an automated colorimetric microbial detection system.

BacT/Alert (Organon Teknika Corp., Durham, N.C.) is an automated microbial detection system based on the colorimetric detection of CO2 produced by growing microorganisms. Results of an evaluation of the media, sensor, detection system, and detection algorithm indicate that the system reliably grows and detects a wide variety of bacteria and fungi. Results of a limited pilot clinical trial with a prototype research instrument indicate that the system is comparable to the radiometric BACTEC 460 system in its ability to grow and detect microorganisms in blood. On the basis of these initial findings, large-scale clinical trials comparing BacT/Alert with other commercial microbial detection systems appear warranted.

Controlled evaluation of 5 versus 10 milliliters of blood cultured in aerobic BacT/Alert blood culture bottles.

Bottles developed for use in the BacT/Alert automated blood culture system (Organon Teknika Corp., Durham, N.C.) can accept up to 10 ml of blood without falling below a 1:5 ratio of blood to broth. We compared the yield and speed of detection of microorganisms in 13,128 adequately filled, paired, aerobic bottles inoculated with 5 versus 10 ml of blood at three university hospitals. A total of 798 microorganisms causing sepsis grew in one or both bottles. The overall recovery of microorganisms from 10-ml samples exceeded that from 5-ml samples (P < 0.001); the increased yield attributed to the additional 5 ml in the samples was 7.2%. The increased yield from 10-ml inocula was most marked for Escherichia coli (P < 0.01) and other members of the family Enterobacteriaceae (P < 0.001). Ten-milliliter samples did not yield more gram-positive bacteria, nonfermentative gram-negative rods, or yeasts. When both bottles were positive, the bottles inoculated with 10 ml of blood showed growth sooner (P < 0.001). Earlier detection with 10-ml inocula was especially notable for coagulase-negative staphylococci (P < 0.001), streptococci (P < 0.001), E. coli (P < 0.025), and other members of the family Enterobacteriaceae (P < 0.025). We conclude that an increase in the volume of blood inoculated into BacT/Alert aerobic blood culture bottles from 5 to 10 ml will increase the overall yield and the speed of detection of clinically important blood pathogens.

Characterization of new biotypes of P157 strain of metarhizium anisopliae var. anisopliae, got by treatment with gamma radiation

Conidios de la cepa silvestre P157 de metarhizium anisopliae var. anisopliae (metsch.) sorokin, fueron expuestos a la radiación gama para la obtención de nuevos biotipos. En la dosis de 390Gy se obtubo 36 colonias (MaP). Cinco colonias (MaP 03, MaP 17, MaP 25, MaP 27), expresaron alteración morfológica para el color en el medio completo. En este, 21 colonias presentaron crecimiento micelial significativamente distinto del testigo silvestre, después de doce días de la inoculación. La colonia MaP 25, presentó mayor valor de crecimiento micelial y la colonia MaP 11, el menor valor. Entre las colonias obtenidas, solamente la colonia MaP28 presentó auxotrofia, identificada como una deficiencia en la síntesis del ácido paraminobenzóico. Los conidios de 29 colonias, presentaron un promedio de porcentaje de germinación significativamente distinto del respectivo aislamiento silvestre, después de doce horas de incubación en medio mínimo, a 25ºC. Las colonias MaP 02, MaP 21 y MaP 36, presentaron precocidad en la germinación de los conidios en relación al patrón silvestre.