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[Blood culture results from 1967 to 1975 and drug sensitivity of bacteria isolated by blood culture in the last three years (1974-1976) (author's transl)].

The blood culture results of patients of S. M. Nuova Hospital of Florence (Italy) have been examined in a period of nine years (1967-1976). We can attest from this results that in the last years of this period we have had an increase of the number of Enterobacteriaceae from blood culture. In the meanwhile we have had a decrease of positive results that is not due to an increase of the number of blood culture; in fact it has been doubled, but this decrease is mostly due to a larger number of examinated patients that to an increase of blood culture for each patient. Introduction in the routine of a diphasic culture system (Castañeda) commercially available has allowed contaminants' decrease. Antibiotic resistance of most important bacteria isolated in the last three years (1974-1976) were carry out.

Anti-Bacterial Agents↗
PubMed1979

Evaluation of ten anaerobic blood culture media.

Selection of an anaerobic blood culture based upon clinical findings that have compared the isolation rates of bacteremic agents from different blood culture media. No agreement has been reached as to which of the commercially available blood culture media is optimal for detection of bacteremia. The purpose of this study was to determine the rates of recovery of anaerobic microorganisms from various anaerobic blood culture media. The blood culture media were inoculated with a small inoculum of microorganisms in the presence or absence of an erythrocyte-serum mixture. The results demonstrated that the type of medium and the erythrocyte-serum mixture influenced the ability of blood culture media to support the growth of microorganisms. The majority of the media failed to support the growth of 87% or more of the microorganisms within four days after inoculation. Pre-reduced brain-heart infusion broth supported the growth of a larger proportion of microorganisms than the other types of blood culture media.

Anaerobiosis↗
PubMed1978

[Further study of the L forms of Streptococci in blood cultures from the patients with rheumatic fever depending on the course of the disease and the methods of cultivation].

Cultures of blood from patients with rheumatism during the active and inactive phases of the disease, treated and untreated with antibiotics were studied. The following results were obtained in blood cultures (on Klodnisky's medium with a 0.01% cystine) of 105 patients with rheumatism: L-forms of streptococcus were isolated in the 88 cases, irrespective of the severity of the disease or the treatment, streptococcus--in 8, and the result proved to be negative in 9 cases. Hemoculture on broth with a 1% glucose solution proved to be positive (streptococcus) in one patient only. The constancy with which the L-forms of streptococcus were isolated in hemocultures from the patients with rheumatism and also the peculiarity of their pathogenic properties modelling the rheumatism under experimental conditions pointed to their role in the pathogenesis of rheumatism. A method of isolation of hemocultures on Klodnitsky's medium with a 0.01% cystine can be recommended for the diagnosis of a septic condition caused by the L-forms of streptococcus in patients suffering from rheumatism.

Adolescent↗
PubMed2025

Whole-genome sequencing-based pathogen characterization for streptococcal infection directly from positive blood culture samples.

Clinical laboratories are increasingly using diagnostic tests directly on positive blood cultures, which may lead to fewer attempts to recover bacterial isolates. Consequently, public health laboratories can benefit from assays that directly process blood culture samples without requiring submission of clinical isolates to determine additional pathogen features not identified by clinical tests, such as vaccine serotype and bacterial genomic relatedness, for surveillance and outbreak response purposes. In partnership with the Minnesota Active Bacterial Core surveillance (ABCs) site, we identified blood culture samples positive for ABCs streptococcal pathogens and characterized them by a direct whole-genome sequencing from blood culture (dWGS) assay. The dWGS results were compared with the results of a reference method (WGS of isolates from the same cultures) to evaluate concordance in pathogen features and genome assemblies. Of the 97 eligible blood culture samples, 83 (86%) passed dWGS quality control criteria and were subjected to a total of 655 dWGS-based tests, which yielded 651 (99.3%) evaluable results. The percent agreement with reference results was 100% (83/83) for M protein gene (emm)/capsular types and 100% (81/81) for multilocus sequencing types. For genotypic antimicrobial susceptibility testing prediction, the percent prediction agreement was 100% (487/487), false resistant prediction rate was 0% (0/417), and the false susceptible prediction rate was 0% (0/66). Assemblies of pathogen genomes from the same patient differed by 1.08 ± 1.68 (mean ± SD) sites per genome. The dWGS assay can extract high-quality, important streptococcal strain characteristics directly from positive blood culture samples to support evolving public health needs.IMPORTANCEWhole-genome sequencing (WGS) technologies have emerged as a transformative toolkit used by public health microbiology laboratories to detect and characterize pathogens. The surveillance of bacterial diseases often relies on clinical laboratories to submit pathogen isolates to regional or national public health laboratories, which have the capacity to routinely conduct WGS-based strain characterization. Clinical laboratories are increasingly using diagnostic tests directly on positive blood cultures, which may lead to fewer attempts to recover bacterial isolates. The study evaluated a direct whole-genome sequencing from blood culture (dWGS) assay that directly processes blood culture samples. The dWGS assay recovered high quality, important streptococcal strain characteristics, including vaccine serotypes and whole-genome assemblies, without requiring submission of clinical isolates. Thus, the dWGS assay represents a promising tool for addressing the evolving needs of public health laboratories in the metagenomics era.

Humans↗
PubMed1979

Counterimmunoelectrophoresis for rapid identification of blood-culture isolates.

A total of 303 blood cultures that were positive by examination of Gram-stained smears were tested immediately by counterimmunoelectrophoresis for detection of bacterial antigens. Antigen was detected in all 82 blood cultures containing Streptococcus pneumoniae and 11 of 22 with Klebsiella pneumoniae, two of two with Haemophilus influenzae, and one of one with Neisseria meningiditis. False-positive cross-reactions in 265 tests occurred only with pneumococcal Omniserum in two cases of nongroupable streptococcal bacteremia and with Klebsiella antiserum in one case of Escherichia coli bacteremia (1.1%). A specific identification of the microorganisms at least 24 hours earlier than by subculture technics was accomplished in 91% of the cultures containing the aforementioned bacteria. The procedure was not useful for detecting antigen in blood cultures containing Staphylococcus aureus.

Antigens, Bacterial↗
PubMed1979

Radiometric detection of Haemophilus in simulated blood cultures.

The radiometric detection of various Haemophilus species was studied in simulated blood cultures and in blood culture media without added blood, but supplemented with nicotinamide-adenine dinucleotide (NAD) or hemin, or both. Haemophilus aphrophilus was the only species studied that was radiometrically detectable without added blood or NAD. All other species studied (Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, and Haemophilus parahaemolyticus) required either added NAD, whole blood, or washed blood cells for growth and radiometric detection. The results of this study and extensive experience with clinical specimens show that the radiometric system is an effective means of rapidly detecting Haemophilus in blood cultures, but it is essential that it be used in conjunction with a subculture three to five days after inoculation.

Bacteriological Techniques↗
PubMed1977

A single-bottle blood culture system: evaluation and comparison with two other systems.

A commercially available single-bottle blood culture system was evaluated at Ben Taub General Hospital, a Harris County District Hospital. Blood cultures from 1010 patients were examined with the Lederle Diagnostics one-bottle blood culture medium-SPS, Columbia broth (E-Vac, Pfizer), and an in-house-prepared brain heart infusion broth with p-aminobenzoic acid (PABA) and 0.1% agar. Of the 1010 patients examined, blood cultures from 211 (20.8%) were positive, yielding a total of 23 different species of microorganisms. Comparison of the results during clinical evaluation, as well as those from simulated blood cultures, showed that the Lederle Diagnostics blood culture bottle was as effective as the in-house-prepared brain heart infusion and commercially available Columbia broths for isolation of aerobes as well as anaerobes. The techniques used in the evaluation and the advantages of a single-bottle culture system are discussed.

Aerobiosis↗
PubMed1977

Development of a lysis-filtration blood culture technique.

A lysed-blood culture system that quickly lyses patients' blood near neutrality and is relatively noninjurious to more delicate pathogens such as Haemophilus influenzae and Bacteroides fragilis is reported. The lysing solution includes culture medium, 0.004 M sodium carbonate and bicarbonate, 0.04% Triton X-100,and 0.6% Rhozyme (a mixture of proteases). Most of the pathogens tested multiplied in the lysing solution. The lysed blood normally is immediately filtered. The membrane is transferred to culture broth. The greatest advantage realized from this blood culture technique is separation of pathogens from antibiotics, bactericidal antibodies, complement, opsonins, and phagocytic systems. Another advantage is the concentration of organisms into a small volume of clear medium for faster growth and visualization of growth. It was observed that both gram-negative and -positive organisms were attracted during filtration to the filter material and were not removed from it by backwashing with buffer. Thus, filter membranes with porosities much larger than would nominally be expected to retain bacteria retained all or part of light and heavy Escherichia coli and Staphylococcus aureus suspensions. Advantage may be taken of this phenomenon to use filters with larger pore sizes and avoid filter clogging by poorly lysed specimens. Porr lysis may result from addition of too much blood to the lysing solution, blood with elevated numbers of erythrocytes or leukocytes, or blood from some people whose blood is naturally more resistant to lysis.

Bacteriological Techniques↗
PubMed1978

Disseminated Nocardia caviae with positive blood cultures.

Disseminated Nocardia caviae infection with multiple positive blood cultures occurred in a bone marrow transplant recipient. Positive blood cultures are unusual in disseminated Nocardia infections and N caviea is an unusual species of Nocardia to cause infections in man, although its virulence in laboratory animals is similar to N asteroides. Multiple positive blood cultures in this case suggest a continuous or recurrent bacteremia rather than a transient bacteremia as previously has been thought to occur in disseminated Nocardia infections. The marked immunosuppressed state of the patient and an indwelling venous line could also have accounted for the recurrent bacteremia.

Adult↗
PubMed1976

A gram stain for smears of blood cultures, body fluids and tissues.

A new Gram stain method for blood cultures was adapted from a method for tissue sections. This was accomplished by comparing various modifications of the latter method on positive smears prepared from a 24-hour negative blood culture seeded with Staphylococcus aureus and Escherichia coli. A blind study was then conducted using the new method and Hucker's method on 24-hour and 72-hour blood cultures as well as a limited number of other body fluids and tissues. Of the 35 positive blood cultures detected by the new method, the Hucker method failed to detect 14 positive specimens (40 per cent) upon initial examination. Ten additional cultures were negative using both stains, but were positive on subculture at three days. The 18 additional cultures positive after ten days were detected using only the Hucker stain.

Acinetobacter↗
PubMed1978

Isolation of Mycoplasma hominis from blood cultures in patients with postpartum fever.

Eight women with postpartum fever are presented in whom Mycoplasma hominis was isolated from cultures of their blood. Clinical disease consisted of a mild although often prolonged febrile illness, and all but one recovered without appropriate antimicrobial therapy. Five patients demonstrated elevated convalescent titers of mycoplasmacidal antibodies. The isolates of M. hominis were recovered from routine blood cultures in the diagnostic bacteriology laboratory using blind subcultures to blood agar plates. These cases lend support to the concept that endometritis with M. hominis is a cause of postpartum fever and suggest that these organisms may be recovered with increased frequency if minor changes in standard bacteriologic technique are introduced.

Bacteriological Techniques↗
PubMed1975

Comparison of recovery rates of various organisms from clinical hypertonic blood cultures by using various concentrations of sodium polyanethol sulfonate.

By using parallel culture techniques, the recovery rates of a wide spectrum of organisms encountered in hypertonic clinical blood cultures was determined from four different blood culture bottles. Each bottle was identical except for the amount of sodium polyanethol sulfonate (SPS) present. Flasks A, B, C, and D contained SPS in final concentrations of 0.025, 0.05, 0.075, and 0%, respectively. Of 144 patients found to have clinically relevant organisms in their blood cultures, 127 had positive A flasks, 144 had positive B flasks, 140 had positive C flasks, and 110 had positive D flasks. There was no significant difference in the time required to obtain organism recovery from the A, B, or C flasks; however, the time required to obtain organism recovery from the D flask was considerably longer, ranging up to 5 days in many cases. Of the various organisms recovered, 3 of 7 strains of anaerobic streptococci and 1 of 28 strains of Streptococcus pneumoniae appeared to be inhibited by SPS when the concentration was 0.075%. In no case was an organism recovered from either the A or D flask but not from the B flask, indicating that a concentration of 0.05% SPS in hypertonic media does not inhibit the growth of a wide spectrum of organisms in clinical blood cultures.

Bacteria↗
PubMed1977

Microcalorimetry as a tool for evaluation of blood culture media.

Evaluation of optimal compositions of blood culture media has called for extensive and laborious work in comparative studies of large series of clincal specimens. Bacterial growth is accompanied by heat production, and calorimetry provides an analytical tool for its detection and quantification. A twin microcalorimeter of the heat conduction type was used to register heat effects in experimentally infected blood cultures. When studying Escherichia coli and Staphylococcus aureus, larger heat effects were produced with 0.05% sodium polyanetholsulfonate than with 600 IU of heparin per ml, which was also the case when using 10% sucrose. The addition of IsoVitaleX (BBL) increased the heat effects produced by the two species mentioned, whereas it had the opposite effect in cultures of Neisseria meningitidis. The present study indicates that microcalorimetry is a valuable and time-saving tool for the evaluation of optimal compositions of bacterial culture media.

Blood↗
PubMed1979

[Importance of skin contamination in blood culture readings].

The importance of the skin contamination for the results of blood cultures was emphasized by model examinations. In the method of blood taking without previous desinfection of the skin the quota of positive blood cultures increased by the twofold to threefold per culture and test person (5.7 to 18.8% and 11.3 to 26.3%, respectively). In large-volume blood takings the contamination rate becomes smaller with increasing blood volume. The rejecting of a first blood sample is to be recommended, when the possibility is given. With an increased quantity o blood per taking by blood bactericidia a decreased contamination rate is to be expected. By the results of the examinations the necessity of a consequent desinfection of the skin is to be emphasized, also when closed systems of blood cultures are used.

Bacteria↗
PubMed2026

Microbial signal profiles and organism-level concordance between plasma metagenomic sequencing and blood culture in suspected bloodstream infection.

Plasma metagenomic next-generation sequencing (mNGS) and blood culture detect different components of the microbial signal and frequently produce discordant organism reports. We characterized microbial signal class, report-derived burden, organism-level concordance, and independent clinical attribution in a retrospective, single-center, episode-level cohort. Among 329 episodes with evaluable plasma mNGS reports, 315 had blood culture performed; 232 were mNGS positive/culture negative and 53 were positive by both methods. In the 232 discordant episodes, the recorded routine-care diagnosis classified 124 as bloodstream infection (BSI) and 108 as non-BSI. Nonviral signals were present in 78.2% and 42.6%, respectively (P&#x2009;<&#x2009;0.001), and median maximum report-derived sequence counts were 98.5 and 11.5 (P&#x2009;<&#x2009;0.001). Two laboratory physicians then independently reviewed source records using structured criteria while masked to the recorded BSI label and mNGS organism and sequence-count information. Initial agreement for the five-category BSI assessment was 97.6% (Cohen's kappa, 0.960). Within the mNGS-positive/culture-negative subgroup, adjudicated BSI likelihood showed a modest ordinal association with report burden (Spearman rho&#x2009;=&#x2009;0.190; P&#x2009;=&#x2009;0.004), while mNGS organisms were considered supported in 1 episode, plausible in 158, unlikely or contaminant in 72, and unresolved in 1. Among 53 dual-positive episodes, 33 (62.3%) shared at least one species, but only 5 (9.4%) had complete species-set concordance. Plasma mNGS and blood culture therefore frequently generated non-equivalent organism sets. Signal class and report burden contributed graded contextual evidence, but organism-level attribution required clinical review and orthogonal microbiology rather than binary positivity alone.

Humans↗
PubMed1979

Standardization of direct susceptibility test for blood cultures.

Insufficient data are available to establish the reliability of direct disk diffusion susceptibility tests performed utilizing positive blood culture broth as inoculum. When Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853 were used, 0.03 ml of turbid overnight blood culture broth was found to produce zone diameters closely approximating the size of diameters obtained by a standardized method. Results of direct (0.03 ml of inoculum) and standardized susceptibility tests were then compared for 116 positive blood cultures (1,069 individual disk comparisons). There were 1,011 test agreements (94.6%). There were also 48 (4.5%) minor discrepancies (change between sensitive and intermediate or between intermediate and resistant) and 10 (0.9%) major discrepancies (change between sensitive and resistant). The major discrepancies were randomly distributed among several organisms and antibiotics. Discrepancies occurred most frequently in the more clinically acceptable direction; i.e., in 79.3% the direct test indicted greater resistance than the standardized test. These data establish that 0.03 ml of turbid overnight blood culture broth produces results which compare closely to those obtained with standard methods, and in practice yield direct susceptibility results with a clinically acceptable level of reliability.

Anti-Bacterial Agents↗
PubMed1979

Recovery of Haemophilus influenzae from twenty-three blood culture media.

Haemophilus influenzae is an important agent of bacteremia and has fastidious growth requirements. The purpose of this investigation was to determine the ability of commercial blood culture media to support the growth of this fastidious microorganism. Twenty-three types of blood culture media were inoculated with individual suspensions of eight strains of H. influenzae in the presence or absence of an erythrocyte-serum mixture. The rates of recovery of the H. influenzae strains from the various types of blood culture media were compared. The results demonstrated that the type of medium, the manufacturer, the erythrocyte-serum mixture, and the strain of H. influenzae influenced the recovery rates of H. influenzae. Optimal recovery of the strains of H. influenzae was obtained from brain heart infustion blood culture medium (GIBCO). Trypic soy broth (GIBCO) and supplemental peptone of Becton, Dickinson and Co. also were found to be superior to the remaining types of media tested for the recovery of H. influenzae.

Blood↗