Comparative evaluation of supplemented peptone broth with sodium polyanetholesulfonate and trypticase soy broth with sodium amylosulfate for detection of septicemia.

We compared the yield and speed of detection of clinically important microorganisms from 10,156 paired 5-ml samples of blood cultured in supplemented peptone broth (SPB) with 0.03% sodium polyanetholesulfonate (SPS) or Trypticase soy broth (TSB) with 0.5% sodium amylosulfate (SAS). The atmosphere of incubation (open venting units) and ratio of blood to broth (1:10) were the same for both samples. Only cultures with adequate blood samples (greater than or equal to 80% of stated volume) were compared statistically. Overall, SPB/SPS outperformed TSB/SAS. Bacteroidaceae and Eubacterium were found more often (P less than 0.05) and viridans streptococci were found sooner (P less than 10(-4)) in SPB/SPS than in TSB/SAS. Most importantly, staphylococci were found both more often (P less than 0.03) and sooner (P less than 10(-7)) in SPB/SPS than in TSB/SAS. In a separate experiment, SAS slowed the growth of a clinical strain of Staphylococcus aureus in TSB. Unless important advantages can be confirmed for SAS in controlled clinical trials, SAS cannot be recommended for routine use as an anticoagulant in blood culture media.     

Controlled evaluation of hypertonic sucrose medium for detection of bacteremia and fungemia in supplemented peptone broth.

Because the value of hypertonic media in detection of bacteremia and fungemia is controversial, we evaluated supplemented peptone broth (SPB) with 0.03% sodium polyanetholsulfonate with and without 10% sucrose in 5,439 paired blood cultures from adult patients. The aerobic atmosphere, 1:10 ratio of blood to broth, and methods for processing blood cultures were identical. Only cultures with adequate blood samples (greater than or equal to 4 ml) were compared statistically. More clinically important bacteria were recovered from SPB with sucrose (P less than or equal to 0.001), including Staphylococcus epidermidis, Enterobacteriaceae, and Bacteroidaceae. However, only one of nine isolates of Neisseria gonorrhoeae grew in SPB with sucrose. Staphylococci (P less than 0.001), Enterobacteriaceae (P less than 0.01), Pseudomonas aeruginosa (P less than 0.01), and yeasts (P less than 0.05) were detected 1 or more days earlier in SPB with sucrose. The effect of sucrose on blood cultures appears to be medium dependent, based on comparisons of our results with those of published reports.     

Controlled evaluation of the effect of atmosphere of incubation on detection of bacteremia and fungemia in supplemented peptone broth.

To evaluate the role of atmosphere of incubation in the detection of clinically important bacteremia and fungemia in adults, we compared the yield of microorganisms from 10,541 paired 5-ml samples of blood incubated aerobically and anaerobically. The medium, supplemented peptone broth (SPB) with 0.03% sodium polyanetholesulfonate, and the ratio of blood to broth (1:10) were the same for all cultures. Only cultures with adequate blood samples (greater than or equal to 80% of stated volume) were compared statistically. More fungi (P less than 10(-7) ) grew in continuously vented bottles of SPB. Aerobic incubation also favored (P less than 0.01) isolation of Neisseria gonorrhoeae and Eubacterium; more than 80% of these bacterial organisms were detected only in vented bottles. Anaerobic incubation (plugged venting units) did not significantly favor the isolation of any genus of microorganisms, although an estimated 11% more Bacteroidaceae grew in the unvented bottle of SPB. By comparison of our data with published results for other media, we conclude that the need for both aerobic and anaerobic incubation of blood cultures is dependent upon the medium used and the microorganisms likely to be encountered. Vented incubation of blood cultured in SPB is crucial for detection of fungi and some bacteria. Routine use of an unvented bottle of SPB may not be worthwhile for patients in whom Bacteroidaceae cause bacteremia infrequently. However, when Bacteroidaceae are suspected as the cause of sepsis, use of an unvented bottle of SPB is prudent.     

Controlled evaluation of hypertonic sucrose medium at a 1:5 ratio of blood to broth for detection of bacteremia and fungemia in supplemented peptone broth.

The value of hypertonic media in the detection of bacteremia and fungemia is controversial, since prior clinical trials have yielded conflicting results with different media. Earlier, we showed that the addition of 10% sucrose to supplemented peptone broth at a 1:10 ratio of blood to broth yielded better recovery of Staphylococcus epidermidis, the Enterobacteriaceae, Pseudomonas aeruginosa, and yeasts. To evaluate the effect of 10% sucrose on blood cultured at a 1:5 ratio, we compared the yield and speed of detection of clinically important microorganisms from adult patients in 5,839 blood samples cultured in supplemented peptone broth with 0.03% sodium polyanetholesulfonate with and without 10% sucrose. The atmosphere of incubation (open venting units), 1:5 ratio of blood to broth, and methods of processing were the same for both bottles. Recovery of facultative gram-positive (P less than 0.02) and gram-negative (P less than 0.02) bacteria was improved, but the recovery of anaerobic gram-negative bacteria was both reduced (P less than 0.01) and delayed (P less than 0.02) by sucrose. The total yield of microorganisms including fungi, however, was increased with sucrose. The effect of sucrose on blood cultures appears to depend on the ratio of blood to broth as well as on the medium used and strains of microorganisms encountered.     

Comparative evaluation of radiometric tryptic soy broth versus radiometric tryptic soy broth with 10% sucrose for detection of bacteremia and fungemia in pediatric patients.

We compared BACTEC radiometric blood culture media with (8B) and without (6B) 10% sucrose for the detection of bacteremia and fungemia in pediatric patients at four university teaching hospitals that used identical methods for obtaining and processing specimens. Overall, the yields of microorganisms from 5,714 blood culture sets were no different in the two media, although a trend was noted favoring 6B for the detection of pneumococci. Speed of detection of positive results was faster in the 6B than in the 8B medium (P less than 0.05), largely due to the faster detection of Staphylococcus aureus in the 6B medium. We conclude that, overall, with pediatric patients the hypertonic 8B radiometric medium has no advantage and that it possibly has a modest disadvantage, compared with isotonic 6B radiometric medium.     

Controlled evaluation of supplemented peptone and Bactec blood culture broths for the detection of bacteremia and fungemia.

The 1-min leukocyte esterase (LE)-nitrite test (Chemstrip 9; Biodynamics, Division of Boehringer Mannheim Biochemicals, Indianapolis, Ind.) and a bioluminescence assay (Monolight centrifugation method; Analytical Luminescence Laboratory, Inc., San Diego, Calif.) were tested for their efficacy as urine screens among 453 patients at a tertiary-care teaching hospital. Both methods had the capacity to exclude significant bacteriuria (greater than or equal to 10(5) CFU/ml) when compared with the results of conventional culture methods, with predictive values of 99 and 93%, respectively, for a negative test. Bioluminescence was the more accurate nonculture method used. Sensitivity and specificity values were 97 and 71%, respectively, for bioluminescence, 82 and 60%, respectively, for LE with nitrite, and 72 and 64%, respectively, for LE without nitrite. At reduced levels of bacteriuria less than 10(5) CFU/ml), the sensitivities of LE-nitrite and bioluminescence were decreased but comparable. The addition of protein and blood test results in the Chemstrip 9, along with LE-nitrite as bacteriuria indicators, were unsatisfactory because of the large numbers of false-positive results attributed to protein and blood determinations. LE activity as detected by the LE test was a poor predictor of significant bacteriuria in both male and female patients. The sensitivity (71%) and specificity (57%) of the LE test in male patients were significantly lower than those previously reported and varied with the patient population studied.     

Controlled evaluation of modified radiometric blood culture medium supplemented with gelatin for detection of bacteremia and fungemia.

Although the addition of 1.2% gelatin to broth blood culture media containing sodium polyanetholesulfonate has been shown to enhance detection of certain bacteria, including Neisseria meningitidis, N. gonorrhoeae, Peptostreptococcus anaerobius, and Gardnerella vaginalis, the effect of such supplementation on the detection of other microorganisms causing bacteremia and fungemia is not known. Therefore, we studied BACTEC 6B medium with and without gelatin in 6,833 paired comparisons to examine the effects of supplementation on both the yield and the speed of detection of sepsis. More aerobic and facultative bacteria grew in the 6B than in the 6B-gelatin medium (P less than 0.001), especially staphylococci (P less than 0.01), Escherichia coli (P less than 0.01), other members of the family Enterobacteriaceae (P less than 0.05), and Acinetobacter spp. (P less than 0.05). When microorganisms grew in both bottles, they did so earlier in 6B than in 6B-gelatin (P less than 0.001). We conclude that the 6B medium in its present formulation is superior to 6B medium supplemented with 1.2% gelatin.     

Clinical comparison of an agar slide blood culture bottle with tryptic soy broth and a conventional blood culture bottle with supplemented peptone broth.

The Roche Septi-Chek biphasic blood culture system with tryptic soy broth was compared with a conventional blood culture bottle with supplemented peptone broth in 6,956 paired blood cultures from adult patients. Both systems were inoculated with equal volumes of blood (5 ml) and incubated aerobically (vented) for 2 weeks. More clinically important bacteria and fungi, including Staphylococcus aureus, S. epidermidis, Escherichia coli and other Enterobacteriaceae, Pseudomonas aeruginosa, and Candida albicans and C. tropicalis were recovered from the biphasic system (P less than 0.001). In contrast, more clinically important anaerobic bacteria (P less than 0.001) and Gardnerella vaginalis (P less than 0.05) were recovered in conventional supplemented peptone broth. Staphylococci (P less than 0.01), Enterobacteriaceae other than E. coli (P less than 0.05), and fungi (P less than 0.001) were detected 1 or more days earlier in the biphasic system, whereas streptococci (P less than 0.001) were detected earlier in the conventional bottle. The overall superiority of the agar slide blood culture system compared with conventional blood culture bottles was confirmed by this evaluation. For optimal detection of anaerobic bacteremia, however, the agar slide bottle should be paired with an anaerobic bottle.     

Tryptophan and hydrogen sulfide reaction from modified trypticase soy agar.

The use of Trypticase soy agar modified by the addition of 0.05% ferric ammonium citrate and 0.01% sodium thiosulfate for the determination of indole, phenylpyruvic acid, anthranilate, kynurenine, and hydrogen sulfide reactions is described.     

Comparison of tryptic soy broth with tryptic soy broth supplemented with sucrose in the septi-chek blood culture system.

The performance of tryptic soy broth (TSB) was compared with TSB supplemented with 10% sucrose (TSB-S) in the Septi-Chek blood culture system (Roche Diagnostics, Div. Hoffmann-La Roche Inc.). A total of 2,292 sets of blood cultures were evaluated. Although slightly more isolates were recovered in the TSB bottle in comparison with the TSB-S bottle, there was no significant difference between the two medium formulations in the recovery of either gram-positive or gram-negative bacteria. Significantly more isolates of fungi were recovered in the TSB bottles. At least 94% of all isolates were detected by day 3 of incubation in both the TSB and TSB-S bottles. Thus, the addition of sucrose to TSB did not result in greater total recovery or faster recovery of the isolates detected in this study.     

Update on detection of bacteremia and fungemia.

The presence of microorganisms in a patient's blood is a critical determinant of the severity of the patient's illness. Equally important, the laboratory isolation and identification of a microorganism present in blood determine the etiologic agent of infection, especially when the site of infection is localized and difficult to access. This review addresses the pathophysiology and clinical characteristics of bacteremia, fungemia, and sepsis; diagnostic strategies and critical factors in the detection of positive blood cultures; characteristics of manual and instrument approaches to bacteremia detection; approaches for isolating specific microorganisms associated with positive blood cultures; and rapid methods for the identification of microorganisms in blood cultures.     

Presumptive identification of Clostridium difficile by detection of p-cresol in prepared peptone yeast glucose broth supplemented with p-hydroxyphenylacetic acid.

Prereduced, anaerobically sterilized peptone yeast glucose broth was supplemented with p-hydroxyphenylacetic acid and used for the presumptive identification of Clostridium difficile. Two hundred eighty-two organisms were grown in this medium for 18 h and tested for p-cresol production by gas-liquid chromatography. All 49 stock and reference strains of C. difficile and 19 organisms confirmed as C. difficile produced p-cresol. p-Cresol was not produced by 53 negative control or 161 test organisms. The system was convenient and effective.     

Comparison of the Oxoid Signal blood culture system with supplemented peptone broth in a pediatric hospital.

We compared the Oxoid Signal bottle (Oxoid, U.S.A.) with supplemented peptone broth (SPB) tubes (B-D Vacutainer; Becton Dickinson Vacutainer Systems) for performing blood cultures in a pediatric hospital. Blood from 3,066 samples was divided equally between the two systems. Of 131 probable pathogens isolated, 121 were detected in the Signal bottle and 111 were detected in the SPB tubes (P greater than 0.05). Of 167 probable contaminants, 122 grew in the Signal bottle and 109 grew in the SPB tubes (P greater than 0.05). The recovery of staphylococci, both probable pathogens and probable contaminants, was increased in the Signal bottle. The recoveries of other organisms, including streptococci, members of the family Enterobacteriaceae, and yeasts, were similar in the two systems. However, the Signal bottle failed to detect three isolates of Haemophilus influenzae, and the time to availability of isolated colonies of other isolates of H. influenzae was delayed. Overall, the Signal bottle was easy and convenient to use, and its innovative detection system should facilitate the early recognition of positive cultures. If its ability to recover H. influenzae can be improved, the Signal bottle could be a useful alternative to existing systems for use in a pediatric setting.     

Comparison of a slide blood culture system with a supplemented peptone broth culture method.

A slide blood culture system (Roche Diagnostics, Div. Hoffman-La Roche, Inc., Montreal, Canada; Roche BCB) was compared with a supplemented peptone broth Vacutainer method (Becton, Dickinson & Co., Rutherford, N.J.) on blood samples taken from the same 1,209 patients. Significantly more clinically important isolates were isolates with the Roche BCB system, and technical processing time was reduced. However, significantly more contaminants were isolated with the Roche BCB system, and it could not readily be adapted for anaerobic culture. Contamination was reduced by careful tightening of the slide to the bottle top to prevent any leakage. Overall, the BCB system is a satisfactory method for aerobic blood culture, markedly reducing technical processing time relative to most other blood culture methods.     

Controlled evaluation of the volume of blood cultured in detection of bacteremia and fungemia.

To evaluate the role of the volume of blood cultured in the detection of clinically important bacteremia and fungemia in adults, we evaluated the yield and speed of detection of microorganisms from 5,317 paired 2- and 5-ml samples of blood. The same kind of medium (supplemented peptone broth with 0.03% sodium polyanetholsulfonate) and atmosphere of incubation (open venting units) were used for all blood cultures. Only adequately filled (less than or equal to 80% of stated volume) sets (20-ml tube and 50-ml bottle) were compared statistically. Significantly more bacteria (p less than 0.01), Pseudomonas spp. In particular (P less than 0.05), were isolated from the 5-ml samples of blood. We conclude that the volume of blood cultured is a critical factor in the detection of septicemia. Consequently, valid evaluation of other factors influencing the detection of septicemia must be based on comparisons in which equal volumes of blood are cultured.     

Controlled evaluation of the agar-slide and radiometric blood culture systems for the detection of bacteremia and fungemia.

A commercially available agar-slide blood culture bottle (Septi-Chek; Roche Diagnostics, Div. Hoffman-La Roche, Inc., Nutley, N.J.) was compared with the radiometric blood culture system (BACTEC; Johnston Laboratories, Inc., Towson, Md.) in 8,544 paired blood cultures from adult patients. The systems were inoculated with equal volumes (10 ml) of blood. Overall, there was no statistically significant difference between the two systems in the recovery of clinically important microorganisms, but significantly more members of the family Enterobacteriaceae other than Escherichia coli were detected by the agar-slide system (P less than 0.005). The agar-slide system detected more fungi, and the BACTEC detected more anaerobic bacteria; however, small numbers of recovered organisms precluded statistical significance. When microorganisms grew in both systems, their presence was detected one or more days earlier in the BACTEC (P less than 0.001). More contaminants grew in the agar-slide system (P less than 0.001). Both systems performed well, and either system should provide high yield and prompt detection of positive blood cultures in patients with bacteremia and fungemia if used in an optimal way as recommended by the respective manufacturers.     

Evaluation of StrepB carrot broth versus Lim broth for detection of group B Streptococcus colonization status of near-term pregnant women

The performance of StrepB Carrot Broth (SCB) versus group B Lim broth (LIM) for detection of group B streptococcus (GBS) colonization status in near-term pregnant women (35 to 37 weeks of gestation) was evaluated. Dually collected vaginal/rectal swabs from 279 women enrolled from a single large maternity clinic were analyzed. Fifty (18%) women were colonized by GBS according to both methods. SCB had excellent diagnostic performance compared to LIM, with sensitivity, specificity, positive predictive value, and negative predictive value of 92%, 100%, 100%, and 98.3%, respectively. Improved diagnostic efficiency due to direct reporting of GBS cases based on an orange color change in the SCB decreased overall labor and material costs.     

Comparative evaluation of Oxoid Signal and BACTEC radiometric blood culture systems for the detection of bacteremia and fungemia.

The Oxoid Signal (Oxoid U.S.A., Inc., Columbia, Md.) blood culture system is a newly described, innovative method for visually detecting growth of microorganisms (D. Sawney, S. Hinder, D. Swaine, and E.Y. Bridson, J. Clin. Pathol. 39:1259-1263, 1986). We did 5,999 paired comparisons of equal volumes (10 ml) of blood in the Oxoid Signal and BACTEC (Johnson Laboratories, Towson, Md.) radiometric blood culture systems at two university hospitals that use identical methods of obtaining and processing specimens. Overall, more microorganisms were detected in the BACTEC system (P less than 0.001), in particular, streptococci (P less than 0.01), fungi (P less than 0.001), and nonfermentative gram-negative rods, especially Acinetobacter species (P less than 0.001). Trends favoring the BACTEC system for detection of Pseudomonas aeruginosa, Haemophilus species, and Neisseria species were noted. There were no differences in the yield of staphylococci, members of the family Enterobacteriaceae, and anaerobic bacteria. When both systems detected sepsis, the BACTEC did so earlier (P less than 0.001). This advantage was most notable at 24 h (70% of BACTEC positives detected versus 48% of Oxoid positives). The proportion of positives detected after 48 h, however, was similar (BACTEC, 84%; Oxoid, 78%). Revisions in the Oxoid Signal system itself or in the processing of Oxoid bottles appear to be necessary to improve its performance in detecting certain microorganism groups, especially fungi.