Sample records for blood culture bottles

  1. Comparison of the BacT/Alert PF Pediatric FAN Blood Culture Bottle with the Standard Pediatric Blood Culture Bottle, the Pedi-BacT

    Krisher, Karen K.; Gibb, Patrick; Corbett, Sandra; Church, Deidre


    The performance of the BacT/Alert PF (Organon-Teknika Corp., Durham, N.C.), a new nonvented pediatric FAN blood culture bottle, was compared to that of the original pediatric bottle, the Pedi-BacT, with matched aerobic cultures obtained from two separate facilities. A total of 244 clinically significant isolates were recovered from 4,015 compliant pairs. Among the positive cultures, 170 (70%) isolates were detected in both the BacT/Alert PF and the Pedi-BacT bottles, while 47 (19%) isolates w...

  2. Value of extended agitation and subculture of BACTEC NR 660 aerobic resin blood culture bottles for clinical yeast isolates.

    Prevost-Smith, E; Hutton, N.


    From 10,351 blood cultures, we prospectively studied 1,000 BACTEC NR 660 aerobic resin blood culture bottles (26+ and Peds Plus) for patients suspected of having yeast septicemia to determine whether extended agitation and subculturing would increase the recovery of yeasts. Aerobic bottles were agitated continuously for 144 h. On day 7, 1,000 culture-negative aerobic bottles which had fungal blood culture requests were agitated for an additional 14 days. During this time they were subcultured...

  3. Improved Diagnosis of Prosthetic Joint Infection by Culturing Periprosthetic Tissue Specimens in Blood Culture Bottles

    Peel, Trisha N.; Dylla, Brenda L.; Hughes, John G.; Lynch, David T.; Greenwood-Quaintance, Kerryl E.; Cheng, Allen C.; Mandrekar, Jayawant N.


    ABSTRACT Despite known low sensitivity, culture of periprosthetic tissue specimens on agars and in broths is routine. Culture of periprosthetic tissue samples in blood culture bottles (BCBs) is potentially more convenient, but it has been evaluated in a limited way and has not been widely adopted. The aim of this study was to compare the sensitivity and specificity of inoculation of periprosthetic tissue specimens into blood culture bottles with standard agar and thioglycolate broth culture, applying Bayesian latent class modeling (LCM) in addition to applying the Infectious Diseases Society of America (IDSA) criteria for prosthetic joint infection. This prospective cohort study was conducted over a 9-month period (August 2013 to April 2014) at the Mayo Clinic, Rochester, MN, and included all consecutive patients undergoing revision arthroplasty. Overall, 369 subjects were studied; 117 (32%) met IDSA criteria for prosthetic joint infection, and 82% had late chronic infection. Applying LCM, inoculation of tissues into BCBs was associated with a 47% improvement in sensitivity compared to the sensitivity of conventional agar and broth cultures (92.1 versus 62.6%, respectively); this magnitude of change was similar when IDSA criteria were applied (60.7 versus 44.4%, respectively; P = 0.003). The time to microorganism detection was shorter with BCBs than with standard media (P < 0.0001), with aerobic and anaerobic BCBs yielding positive results within a median of 21 and 23 h, respectively. Results of our study demonstrate that the semiautomated method of periprosthetic tissue culture in blood culture bottles is more sensitive than and as specific as agar and thioglycolate broth cultures and yields results faster. PMID:26733067

  4. Validity of direct identification and antibiotic susceptibility of microrganisms from bottles of blood culture

    Carmela Mazzone


    Full Text Available The blood culture is a very important laboratory test: if bacteremia or sepsis are suspected, the diagnosis of the pathogen and antibiotic therapy may be achieved making use of it. Identification and antibiotic susceptibility test carried out directly from the bottle may give important information in a shorter time. The introduction of the automatic instrumentation has improved the discovering of pathogens in the blood, however the elapsing time between the positive detection and the microbiological report is still along.The aim of our work was to verify the validity of the direct use of blood culture broth in which growth of microorganisms has been detected, which could reduce the response time of the bacteremia diagnosis. During the period February - July 2009, a total of 150 blood cultures were analysed:we compared the results obtained both by direct method and by reference method. 20 Gram positive microrganisms and 13 Gram negative microrganisms were respectively isolated and identified. The identification of Gram-negative and Gram-positive microrganisms showed an agreement of 100% between the direct and the reference method. For antibiotic susceptibility tests, among the Gram positive has reported 1.3% very major error, 2.9% major error and 1.4% minor error, while the Gram negative, respectivety 0.3%, 1.4%, 0%. The use of direct identification and susceptibility testing from positive blood cultures, can improve the response time and better efficiency in diagnostic procedures.


    A new fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for identification of Candida albicans directly from yeast-positive blood culture bottles is described. The test (C. albicans PNA FISH) is based on a fluorescein-labeled PNA probe targeting C. albicans 26...

  6. Improved Diagnosis of Orthopedic Implant-Associated Infection by Inoculation of Sonication Fluid into Blood Culture Bottles

    Portillo, María Eugenia; Salvadó, Margarita; Trampuz, Andrej; Siverio, Ana; Alier, Albert; Sorli, Lluisa; Martínez, Santos; Pérez-Prieto, Daniel; Horcajada, Juan P.; Puig-Verdie, Lluis


    Sonication improved the diagnosis of orthopedic implant-associated infections (OIAI). We investigated the diagnostic performance of sonication fluid inoculated into blood culture bottles in comparison with that of intraoperative tissue and sonication fluid cultures. Consecutive patients with removed orthopedic hardware were prospectively included and classified as having OIAI or aseptic failure (AF) according to standardized criteria. The diagnostic procedure included the collection of five i...

  7. Improved diagnosis of orthopedic implant-associated infection by inoculation of sonication fluid into blood culture bottles

    Portillo, Mar??a Eugenia; Salvad??, Margarita; Trampuz, Andrej; Siverio, Ana; Alier, Albert; Sorli Red??, M. Luisa; Mart??nez, Santos; P??rez, Daniel; Horcajada Gallego, Juan Pablo; Puig Verdi??, Lu??s


    Sonication improved the diagnosis of orthopedic implant-associated infections (OIAI). We investigated the diagnostic performance of sonication fluid inoculated into blood culture bottles in comparison with that of intraoperative tissue and sonication fluid cultures. Consecutive patients with removed orthopedic hardware were prospectively included and classified as having OIAI or aseptic failure (AF) according to standardized criteria. The diagnostic procedure included the collection of five i...

  8. Blood Culture Test

    ... difficult to grow in culture, and additional blood cultures using special nutrient media may be done to try to grow and identify the pathogen . Viruses cannot be detected using blood culture bottles designed to grow bacteria. If the health ...

  9. Improved method of isolating bacteria from joint fluids by the use of blood culture bottles.

    von Essen, R; Hölttä, A


    An analysis of 47 episodes of bacterial arthritis showed that applying a blood culture procedure to the culture of joint fluids gave positive results in 10 cases (21%) that were negative by conventional methods. When follow up samples taken during antibiotic treatment were considered the proportion of false negatives eliminated rose to 40%. The respective advantages of using a large volume of inoculum and a large volume of medium are that very low numbers of viable bacteria in infected fluid ...

  10. Rapid Detection of blaKPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles

    Hindiyeh, Musa; Smollan, Gill; Grossman, Zehava; Ram, Daniela; Robinov, Jana; Belausov, Natasha; Ben-David, Debbie; Tal, Ilana; Davidson, Yehudit; Shamiss, Ari; Mendelson, Ella; Keller, Nathan


    Rapid detection of drug-resistant bacteria in clinical samples plays an instrumental role in patients' infection management and in implementing effective infection control policies. In the study described in this report, we validated a multiplex TaqMan real-time quantitative PCR (qPCR) assay for the detection of blaKPC genes and the human RNase P gene in Bactec blood culture bottles. The MagNA Pure LC (version 2.0) instrument was utilized to extract nucleic acids from the inoculated broth, wh...

  11. Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry.

    Bernard La Scola

    Full Text Available BACKGROUND: With long delays observed between sampling and availability of results, the usefulness of blood cultures in the context of emergency infectious diseases has recently been questioned. Among methods that allow quicker bacterial identification from growing colonies, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF mass spectrometry was demonstrated to accurately identify bacteria routinely isolated in a clinical biology laboratory. In order to speed up the identification process, in the present work we attempted bacterial identification directly from blood culture bottles detected positive by the automate. METHODOLOGY/PRINCIPAL FINDINGS: We prospectively analysed routine MALDI-TOF identification of bacteria detected in blood culture by two different protocols involving successive centrifugations and then lysis by trifluoroacetic acid or formic acid. Of the 562 blood culture broths detected as positive by the automate and containing one bacterial species, 370 (66% were correctly identified. Changing the protocol from trifluoroacetic acid to formic acid improved identification of Staphylococci, and overall correct identification increased from 59% to 76%. Lack of identification was observed mostly with viridans streptococci, and only one false positive was observed. In the 22 positive blood culture broths that contained two or more different species, only one of the species was identified in 18 samples, no species were identified in two samples and false species identifications were obtained in two cases. The positive predictive value of bacterial identification using this procedure was 99.2%. CONCLUSIONS/SIGNIFICANCE: MALDI-TOF MS is an efficient method for direct routine identification of bacterial isolates in blood culture, with the exception of polymicrobial samples and viridans streptococci. It may replace routine identification performed on colonies, provided improvement for the specificity of blood culture

  12. High medical impact of implementing the new polymeric bead-based BacT/ALERT® FAPlus and FNPlus blood culture bottles in standard care.

    Amarsy-Guerle, R; Mougari, F; Jacquier, H; Oliary, J; Benmansour, H; Riahi, J; Berçot, B; Raskine, L; Cambau, E


    Blood culture (BC) efficiency is critical for the diagnosis of bloodstream infection (BSI). We evaluated the impact on standard care of implementing the new BacT/ALERT® FAPlus and FNPlus BC bottles containing antibiotic-binding polymeric beads. We measured positivity rates and time to detection (TTD) during the first 10 months of implementation (PF) and during the previous 10-month period (PS) during which we were using standard aerobic (SA) or standard anaerobic (SN) BC bottles. For each period, the same number of consecutive patients (n = 3,918) was included. Per patient, a median of 1 BC set (1 aerobic and 1 anaerobic bottles) has been sampled. A higher positivity rate was measured during PF than PS when counting per BC bottle (7.0 % vs 5.8 % with 1,456 and 1,237 positive bottles respectively, P < 0.0001) and per BC set (9.6 % vs 7.8 % with 995 and 832 positive BC sets respectively, P < 0.0001). In PF, an increased number of cases due to staphylococci (P < 0.0001) and to Gram-negative bacilli (P < 0.005) was observed, whereas the contamination rate was similar during the two periods (2.4 % of BC sets in PF and 2.3 % in PS). Although antibiotic consumption and medical activity were similar during the two periods, BSI case detection increased from 2.2 to 2.6 per 1,000 hospital-days, especially in intensive care units (ICU; 35.1 to 55.7). Mean TTD for pathogenic microorganisms was significantly shorter in PF than in PS (15.5 h vs 18.0 h, P < 0.01). In conclusion, the use of the new FAPlus/FNPlus BC bottles improved the diagnosis of bacteremia in our hospital, especially in ICU patients. PMID:25648261

  13. Development of a rapid diagnostic method for identification of Staphylococcus aureus and antimicrobial resistance in positive blood culture bottles using a PCR-DNA-chromatography method.

    Ohshiro, Takeya; Miyagi, Chihiro; Tamaki, Yoshikazu; Mizuno, Takuya; Ezaki, Takayuki


    Blood culturing and the rapid reporting of results are essential for infectious disease clinics to obtain bacterial information that can affect patient prognosis. When gram-positive coccoid cells are observed in blood culture bottles, it is important to determine whether the strain is Staphylococcus aureus and whether the strain has resistance genes, such as mecA and blaZ, for proper antibiotic selection. Previous work led to the development of a PCR method that is useful for rapid identification of bacterial species and antimicrobial susceptibility. However, that method has not yet been adopted in community hospitals due to the high cost and methodological complexity. We report here the development of a quick PCR and DNA-chromatography test, based on single-tag hybridization chromatography, that permits detection of S. aureus and the mecA and blaZ genes; results can be obtained within 1 h for positive blood culture bottles. We evaluated this method using 42 clinical isolates. Detection of S. aureus and the resistance genes by the PCR-DNA-chromatography method was compared with that obtained via the conventional identification method and actual antimicrobial susceptibility testing. Our method had a sensitivity of 97.0% and a specificity of 100% for the identification of the bacterial species. For the detection of the mecA gene of S. aureus, the sensitivity was 100% and the specificity was 95.2%. For the detection of the blaZ gene of S. aureus, the sensitivity was 100% and the specificity was 88.9%. The speed and simplicity of this PCR-DNA-chromatography method suggest that our method will facilitate rapid diagnoses. PMID:27056092

  14. Blood culture

    Culture - blood ... A blood sample is needed . The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

  15. Comparative Analysis of Two Broad-Range PCR Assays for Pathogen Detection in Positive-Blood-Culture Bottles: PCR–High-Resolution Melting Analysis versus PCR-Mass Spectrometry

    Jeng, Kevin; Gaydos, Charlotte A; Blyn, Lawrence B.; Yang, Samuel; Won, Helen; Matthews, Heather; Toleno, Donna; Hsieh, Yu-Hsiang; Carroll, Karen C.; Hardick, Justin; Masek, Billy; Kecojevic, Alexander; Sampath, Rangarajan; Peterson, Stephen; Rothman, Richard E.


    Detection of pathogens in bloodstream infections is important for directing antimicrobial treatment, but current culture-based approaches can be problematic. Broad-range PCR assays which target conserved genomic motifs for postamplification amplicon analysis permit detection of sepsis-causing pathogens. Comparison of different broad-range assays is important for informing future implementation strategies. In this study, we compared positive-blood-culture bottles processed by PCR coupled to hi...

  16. A cost-saving algorithm for rapid diagnosis of Staphylococcus aureus and susceptibility to oxacillin directly from positive blood culture bottles by combined testing with BinaxNOW® S. aureus and Xpert MRSA/SA Assay.

    Yossepowitch, Orit; Dan, Michael; Kutchinsky, Anuta; Gottesman, Tamar; Schwartz-Harari, Orna


    We studied an algorithm combining 2 rapid methods to detect Staphylococcus aureus and its susceptibility to oxacillin directly from positive blood cultures; our goal was to reduce the cost of the procedure, while maintaining accuracy and a short turnaround time. A total of 581 blood cultures containing gram-positive cocci in clusters were tested by BinaxNOW® Staphylococcus aureus Test. Positive samples were further assessed by the Xpert MRSA/SA BC Assay. Phenotypic methods have identified coagulase-negative staphylococci in 505 samples and S. aureus in 76 samples, of which 51 were oxacillin sensitive and 25 were oxacillin resistant. Sensitivity and specificity of the BinaxNOW® Test were 92% and 99%, respectively, compared to the phenotypic method. The Xpert MRSA/SA BC Assay showed complete concordance with phenotypic identification and antimicrobial susceptibility results. The combined rapid assays produced results within 2 hours and reduced the cost by 75% compared with the Xpert MRSA/SA BC Assay if used alone on all blood bottles. PMID:24503507

  17. Routine aerobic terminal subculturing of blood cultures in a cancer hospital.

    Kiehn, T E; Wong, B; Edwards, F F; Armstrong, D.


    Routine terminal aerobic subcultures of macroscopically negative blood culture bottles were evaluated during a 15-month period when 30,000 blood cultures were processed. Each blood culture set consisted of a vented and an unvented 50-ml broth bottle. Forty-eight pathogens and 47 contaminants were isolated only from terminal subcultures. Twenty-two of the significant isolates were yeasts (usually recovered from vented bottles), and 10 were Pseudomonas aeruginosa (usually recovered from unvente...

  18. Evaluation of blood culture media for isolation of pyridoxal-dependent Streptococcus mitior (mitis).

    Gross, K. C.; Houghton, M P; Roberts, R. B.


    Nutritional variant streptococci identified as pyridoxal-dependent Streptococcus mitior (mitis) account for 5 to 6% of streptococcal endocarditis and may be a cause of "culture-negative" endocarditis. Hence, growth of three variant strains in 11 commercial blood culture broths was compared to that in fresh heart infusion broth. For simulation of clinical specimens, culture bottles were injected with 5 ml of human blood, inoculated with approximately 500 colony-forming units (CFU) per bottle, ...

  19. Blood Culture (For Parents)

    ... KidsHealth in the Classroom What Other Parents Are Reading Upsetting News Reports? What to Say Vaccines: Which ... BMP) Blood Test: Complete Blood Count Basic Blood Chemistry Tests Getting a Blood Test (Video) Blood Test: ...

  20. Detection of bacteremia by Difco ESP blood culture system.

    Morello, J A; Leitch, C; Nitz, S.; Dyke, J W; Andruszewski, M; Maier, G.; Landau, W.; Beard, M. A.


    In a multicenter study, the Difco ESP blood culture system (Difco Laboratories, Detroit, Mich.) was compared with the BACTEC NR660 system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.). The ESP system monitors each blood culture bottle every 12 to 24 min to detect changes in oxygen consumption and gas production by microbes. Equal volumes of blood were inoculated into aerobic ESP-80A and BACTEC 6A, 16A, or PEDS Plus broths and anaerobic ESP-80N and BACTEC 7A or 17A broths and w...

  1. Clinical laboratory comparison of the 10-ml isolator blood culture system with BACTEC radiometric blood culture media.

    Kellogg, J A; Manzella, J P; McConville, J H


    The efficiency of the 10-ml Isolator (E. I. du Pont de Nemours & Co., Inc.) for recovery of pathogens from blood was compared with that of BACTEC 6B and 7C media (Johnston Laboratories) by using 4,195 cultures from 1,662 patients. During the first phase of the study, BACTEC bottles were inoculated with 3 ml of blood; in the second phase, bottles were inoculated with 5 ml. The objectives were to compare results with similar blood volumes used for the detection of anaerobes as well as similar overall volumes and to determine the relative sensitivity of BACTEC media inoculated with the minimum and maximum volumes suggested by the manufacturer. From 180 patients, 391 significant isolates were recovered, 354 (91%) with the Isolator and 304 (78%) with the bottles. Isolators recovered 31 (15%) and 19 (18%) more pathogens overall than did the two-bottle system inoculated with 3 and 5 ml of blood, respectively, including 30 (36%) and 10 (34%) more Enterobacteriaceae. Recovery of anaerobes was greater in the BACTEC anaerobic medium, but only when its inoculum was increased to 5 ml. No significant differences existed between the two systems in pathogen detection times or detection of polymicrobic bacteremia. The Isolator contamination rate (8.3%) was approximately 4 times that of the bottles. The number of CFU of pathogen per milliliter of blood, blood volume sampled, and number of Isolators collected were more important than antimicrobial agent pretreatment in contributing to patient bacteremia of fungemia undetected by the Isolator. The Isolator appeared to be a practical alternative for recovery of aerobic and facultatively anaerobic pathogens from the blood. PMID:6386871

  2. Classification of positive blood cultures

    Gradel, Kim Oren; Knudsen, Jenny Dahl; Arpi, Magnus;


    ABSTRACT: BACKGROUND: Information from blood cultures is utilized for infection control, public health surveillance, and clinical outcome research. This information can be enriched by physicians assessments of positive blood cultures, which are, however, often available from selected patient groups...... or pathogens only. The aim of this work was to determine whether patients with positive blood cultures can be classified effectively for outcome research in epidemiological studies by the use of administrative data and computer algorithms, taking physicians assessments as reference. METHODS: Physicians...... assessments of positive blood cultures were routinely recorded at two Danish hospitals from 2006 through 2008. The physicians assessments classified positive blood cultures as: a) contamination or bloodstream infection; b) bloodstream infection as mono- or polymicrobial; c) bloodstream infection as community...

  3. Blood cultures in ambulatory outpatients

    Laupland Kevin B


    Full Text Available Abstract Background Blood cultures are a gold standard specific test for diagnosing many infections. However, the low yield may limit their usefulness, particularly in low-risk populations. This study was conducted to assess the utility of blood cultures drawn from ambulatory outpatients. Methods Blood cultures drawn at community-based collection sites in the Calgary Health Region (population 1 million in 2001 and 2002 were included in this study. These patients were analyzed by linkages to acute care health care databases for utilization of acute care facilities within 2 weeks of blood culture draw. Results 3102 sets of cultures were drawn from 1732 ambulatory outpatients (annual rate = 89.4 per 100,000 population. Significant isolates were identified from 73 (2.4% sets of cultures from 51 patients, including Escherichia coli in 18 (35% and seven (14% each of Staphylococcus aureus and Streptococcus pneumoniae. Compared to patients with negative cultures, those with positive cultures were older (mean 49.6 vs. 40.1 years, p Conclusion Blood cultures drawn in outpatient settings are uncommonly positive, but may define patients for increased intensity of therapy. Strategies to reduce utilization without excluding patients with positive cultures need to be developed for this patient population.

  4. Survey on Heterotrophic Bacterial Contamination in Bottled Mineral Water by Culture Method

    Essmaeel Ghorbanalinezhad


    Full Text Available Background and Aim: This project focuses on the level of heterotrophic baceria in bottled mineral water which could be a health concern for the elderly, infants, pregnant women and immuno-compromised patients. Materials and Methods: Different brands of bottled water samples were selected randomly and evaluated for their bacteriological quality, using different specific culture media and biochemical tests. Water samples were analyzed within 24 hours of their purchase/collection. Samples were filtered with 0.45 micron and filters were plated in different media. Then media were incubated at 37˚C for 24-48 hours. Results: Morphological study and biochemical tests revealed a number of bacteria in different   brands of  bottled water. Heterotrophic bacteria(Gram positive cocci, Spore forming gram positive bacilli, non spore forming gram positive bacilli, gram negative bacilli, and gram negative coccobacilli; Pseudomonas and Stenotrophomonas counted in 70% of bottled water samples. There were no cases of fecal contamination or the presence of E.coli. Conclusions: Bottled water is not sterile and contains trace amounts of bacteria naturally present or introduced during processing. Testing drinking water for all possible pathogens is complex, time-consuming, and expensive. If only total coliform bacteria are detected in drinking water, the source is probably environmental. Since the significance of non-pathogenic heterotrophic bacteria in relation to health and diseases is not understood, there is an urgent need to establish a maximum limit for the heterotrophic count in the bottled mineral water. Growth conditions play a critical role in the recovery of heterotrophic bacteria in bottled drinking water.

  5. Rapid organism identification from Bactec NR blood culture media in a diagnostic microbiology laboratory.

    Claxton, P M; Masterton, R G


    AIMS--To evaluate rapid organism identification on positive blood culture Bactec NR media (phial types 26, 27, 42 and 17), and to assess the usefulness of these procedures in a diagnostic microbiology laboratory. METHODS--Two hundred and sixty, first positive, blood culture bottles from individual patients were tested by rapid identification methods selected on the basis of Gram film organism morphology. Tube coagulase and latex agglutination were applied to presumptive staphylococci; latex a...

  6. Simple spinner bottle with rotating basket packed with carriers for hybridoma cell culture.

    Chen, Y; Wang, G; Zhang, W; Freedman, D


    r-69B is a mouse-mouse hybridoma cell line, producing monoclonal antibody IgG against human r-IFN. It was cultured for 21 days in the 1.0-L spinner bottle which was assembled with a rotating basket packed with the 8.0-g Fibra-Cel carriers. The agitation was 100 r/min. The results showed that 53.5% of the cells can be trapped within the carriers in the basket and the cell concentration and MAb was about double those in the suspension culture. The spinner bottle could be assembled simply and used in general laboratories. It also could be used for different kinds of cells, including anchorage-dependent and independent cells. PMID:9093764

  7. Use of BBL CHROMagar MRSA Medium for Identification of Methicillin-Resistant Staphylococcus aureus Directly from Blood Cultures

    Pape, John; Wadlin, Jill; Nachamkin, Irving


    We evaluated the ability of BBL CHROMagar MRSA medium (Becton Dickinson, Sparks, MD) to identify methicillin-resistant Staphylococcus aureus (MRSA) directly upon subculture from positive blood culture bottles. There were 124 MRSA isolates recovered from blood cultures in the study. BBL CHROMagar MRSA medium was highly sensitive (97.6% [121/124] at 18 to 24 h of incubation and 100% [124/124] at 48 h) and 99.9% specific for identifying MRSA from positive blood cultures.

  8. Rapid identification of Staphylococcus aureus and Streptococcus pneumoniae from blood cultures.

    Knight, R G; Shlaes, D M


    Simultaneous application of the lysostaphin sensitivity test for identification of Staphylococcus aureus and the deoxycholate test for the identification of Streptococcus pneumoniae was evaluated for reliability in rapid identification (1 h) of these organisms from blood cultures by using BACTEC 6B and 7C bottles. The procedure was applied to 127 cultures, 74 lysostaphin tests and 53 deoxycholate tests. Lysostaphin-tested organisms included 23 S. aureus, 40 Staphylococcus epidermidis, 1 Liste...

  9. Acridine orange staining and radiometric detection of microorganisms in blood cultures

    To determine whether acridine orange (AO) staining of blood cultures could be used as a substitute for blind subculture when used in conjunction with the BACTEC system (Johnston Laboratories, Inc., Towson, Md.), the two methods were compared on all BACTEC-negative specimens. Since blind subcultures were routinely performed in our laboratory on days 2 and 6 of incubation, AO staining was also performed on these days. Cultures which were BACTEC positive on day 1 of incubation were not included in the study. Of the 2,395 bottles tested after 2 days of incubation, 106 were subculture positive. Of these, 96 (90.6%) were also AO positive and BACTEC positive, 3 (2.8%) were AO positive and BACTEC negative, and 7 (6.6%) were AO negative and BACTEC positive. Of the 3,487 bottles tested on day 6 of incubation, 14 were subculture positive; 7 (50%) of these were AO positive and BACTEC positive, and seven were AO positive and BACTEC negative. Of the total of 10 culture-positive bottles missed by BACTEC, all were positive, and all 10 companion aerobic bottles were BACTEC positive. In both phases of the experiment, there was a total of only four false-positive AO stains. As a result of this investigation, we have substituted AO staining for blind subculturing of BACTEC-negative bottles

  10. Use of MALDI-TOF MS technique for rapid identification of bacteria from positive blood cultures

    Sung Kuk Hong


    Full Text Available We evaluated the feasibility of same-day routine aerobic bacterial identification using the following procedures: Picking colonies from 4 and 6 h incubated subculture from positive blood culture bottle and analyzing them by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS. The matched identification rate of this procedure at the species level was 80.6% (141/175 for the 4-h cultures compared with overnight cultures and 90.9% (159/175 for the 6-h cultures. Thus, our technique provides an easy and rapid method for identification of aerobic bacteria in routine clinical microbiology laboratories.

  11. Evaluation of Verigene Blood Culture Test Systems for Rapid Identification of Positive Blood Cultures

    Jae-Seok Kim; Go-Eun Kang; Han-Sung Kim; Hyun Soo Kim; Wonkeun Song; Kyu Man Lee


    The performance of molecular tests using the Verigene Gram-Positive and Gram-Negative Blood Culture nucleic acid tests (BC-GP and BC-GN, resp.; Naosphere, Northbrook, IL, USA) was evaluated for the identification of microorganisms detected from blood cultures. Ninety-nine blood cultures containing Gram-positive bacteria and 150 containing Gram-negative bacteria were analyzed using the BC-GP and BC-GN assays, respectively. Blood cultures were performed using the Bactec blood culture system (BD...

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

    Reimer, L G; McDaniel, J D; Mirrett, S.; Reller, L B; Wang, W L


    Comparison of conventional blood culture media with newer formulations of Bactec media for radiometric detection are lacking. Therefore, we compared the yield and speed of detection of clinically important microorganisms with supplemented peptone broth (SPB) and Bactec aerobic (6B) and anaerobic (7C or 7D) broths in 7,627 blood samples from adult patients. Acridine orange stains from SPB, radiometric readings from Bactec, and routine subcultures from all bottles were done at the same time int...

  13. Advantage of combining resin with lytic BACTEC blood culture media.

    Rohner, P; Pepey, B; Auckenthaler, R


    The BACTEC 9240 (Becton Dickinson, Sparks, Md.) automated blood culture system is based on the continuous monitoring of CO2 production by means of a fluorescent sensor attached to the bottom of a culture vial. We compared two media for this system, resin-containing Plus aerobic/F and Lytic anaerobic/F. Sets of Plus aerobic/F and Lytic anaerobic/F vials inoculated with similar volumes (9 +/- 2.5 ml) were evaluated. In the laboratory, the vials were introduced into the system in accordance with the recommendations of the manufacturer and incubated at 35 degrees C for 5 days. A total of 10,914 sets consisting of two bottles each were obtained from 3,674 patients (2.97 cultures per patient). Of these, 1,233 (11%) were culture positive, including 1,074 (10%) yielding at least one pathogen, and 178 (2%) were contaminated. A total of 1,135 isolates were considered clinically relevant in 624 septic episodes; we isolated 894 from Plus aerobic/F and 852 from Lytic anaerobic/F (P = 0.06 [not significant]). More S. aureus isolates (P = 0.05), Pseudomonas spp. (P aerobic/F medium, but more streptococci (P aerobic/F vials (n = 112 versus 52, respectively). Significantly more (P aerobic/F vials (n = 210; 1.9%) than Lytic anaerobic/F vials (n = 42; 0.4%) were unconfirmed positives. Plus aerobic/F and Lytic anaerobic/F proved to be a valuable pair of blood culture media. Plus aerobic/F performs better for patients under antibiotic treatment, due to the antimicrobial-neutralizing effect of resins. For patients without antibiotic therapy, more microorganisms could be isolated from Lytic anaerobic/F due to cell lysis. PMID:9316921

  14. Impact of systemic antifungal therapy on the detection of Candida species in blood cultures in clinical cases of candidemia.

    Bailly, S; Garnaud, C; Cornet, M; Pavese, P; Hamidfar-Roy, R; Foroni, L; Boisset, S; Timsit, J-F; Maubon, D


    The diagnosis and follow-up of candidemia still rely on blood cultures (BCs). In vitro studies show that antifungals can significantly modify the result of blood culture not containing adsorbing agents. We aimed to evaluate, under clinical conditions, the impact on BC yeast detection of systemic antifungal therapy (SAT). Patients (n = 125) experiencing candidemia at Grenoble University Hospital (France) were included in a 4-year retrospective study. The Plus Aerobic/F (Aerobic) and Plus Anaerobic/F (Anaerobic) bottles, which both contain adsorbing resins and the non-resin selective Mycosis IC/F (Mycosis) bottles, were compared using multivariate hierarchical models adjusted for clinical characteristics. The positivity rate (PR) is decreased in patients with SAT (p SAT reduces PR by a factor of 0.16 (95 % CI: [0.08; 0.32]) and increases the time to positivity (TTP) by a factor of 1.76 ([1.30; 2.40]; p SAT, TTP is higher in non-resin bottles (Mycosis) than in resin bottles (RR = 1.76, [1.30; 2.40]); however, the TTP in nonresin and resin bottles remains comparable. Although discordant results are observed with and without SAT (37 and 58 % respectively), we showed that the presence of SAT decreases significantly the agreement rate by a factor of 0.29 (CI: [0.12; 0.68]). The combination of Anaerobic and Mycosis bottles allowed a 100 % positivity rate for C. glabrata. SAT significantly affects BC results. Because they provide additional and complementary results, this study supports the concomitant use of resin and selective bottles, especially in patients receiving SAT. PMID:27039341

  15. Reliability of direct sensitivity determination of blood cultures

    The aim of this study was to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. All blood culture samples received at Microbiology Laboratory from 1st July 2006 to 31st August 2006 were ncluded in the study. All samples were inoculated in automated blood culture system BACTEC 9240 which contained enriched Soybean-Casein Digest broth with CO/sub 2/. Once positive, bottles were removed from system; gram staining of the positive broths was done. Susceptibility test was performed from positive broth, on MHA (Mueller-Hinton Agar), with antibiotics panel according to gram stain result. All positive broths were also sub-cultured on blood agar, chocolate agar and McConkey agar for only gram-negative rods. Next day, the zone sizes of all antibiotics were recorded using measuring scale and at the same time susceptibility test was repeated from isolated colonies from subcultures, with inoculums prepared of McFarland 0.5 standard 0.2 Staphylococcus aureus (ATCC 29213); E.coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853) were included as quality control strain. Zone sizes were interpreted as sensitive (S), resistant (R) and intermediate (I) according to CLSI recommendation. Two results were compared and recorded. Out of a total 1083 combinations, zone diameters by standard method were either equal or greater than direct zone diameter (never smaller). Most of the discrepancies were in b-lactam/b-lactamase combinations and aminoglycosides. While reporting these groups of antibiotics with direct sensitivity test, one should be cautious. These are the major antibiotic used for life-threatening infections. In case of being heavy/lighter standard inoculums or marginal zones, repeating with standard method should be preferred to minimize the chances of error. (author)

  16. Evaluation of Verigene Blood Culture Test Systems for Rapid Identification of Positive Blood Cultures

    Jae-Seok Kim


    Full Text Available The performance of molecular tests using the Verigene Gram-Positive and Gram-Negative Blood Culture nucleic acid tests (BC-GP and BC-GN, resp.; Naosphere, Northbrook, IL, USA was evaluated for the identification of microorganisms detected from blood cultures. Ninety-nine blood cultures containing Gram-positive bacteria and 150 containing Gram-negative bacteria were analyzed using the BC-GP and BC-GN assays, respectively. Blood cultures were performed using the Bactec blood culture system (BD Diagnostic Systems, Franklin Lakes, NJ, USA and conventional identification and antibiotic-susceptibility tests were performed using a MicroScan system (Siemens, West Sacramento, CA, USA. When a single strain of bacteria was isolated from the blood culture, Verigene assays correctly identified 97.9% (94/96 of Gram-positive bacteria and 93.8% (137/146 of Gram-negative bacteria. Resistance genes mecA and vanA were correctly detected by the BC-GP assay, while the extended-spectrum β-lactamase CTX-M and the carbapenemase OXA resistance gene were detected from 30 cases cultures by the BC-GN assay. The BC-GP and BC-GN assays showed high agreement with conventional identification and susceptibility tests. These tests are useful for rapid identification of microorganisms and the detection of clinically important resistance genes from positive Bactec blood cultures.

  17. Utilization of blood cultures in Danish hospitals

    Gubbels, S; Nielsen, J; Voldstedlund, M;


    This national population-based study was conducted as part of the development of a national automated surveillance system for hospital-acquired bacteraemia and ascertains the utilization of blood cultures (BCs). A primary objective was to understand how local differences may affect interpretation...

  18. Sepsis: a more faster report from blood culture

    Raffaella Ledonne


    Full Text Available Introduction: Sepsis is a leading cause of morbidity and mortality. Several studies demonstrated that the earliness of the intervention therapy, including antimicrobial treatment active on the specific pathogen, is associated with a reduction of mortality. In order to permit the use of a quicker appropriate treatment in respect to using the conventional method,we evaluated both the accuracy of strain identification and in vitro antimicrobial susceptibilities directly from the haemocultural bottles. Methods:We examined 112 samples of positive blood cultures through traditional technique of subculture, identification and susceptibility testing, and a diagnostic alternative method in the following way: 5-6 ml from positive blood cultures (BACTEC 9640 were transferred into a test tube fitted with a sterile gel separator and centrifuged at 3200 rpm for 15 minutes. After the removal of supernatant, the microbial pellet was suspended in a saline buffer to obtain an 0.5 McFarland inoculum; this turbidity was needed to set up the direct identification and the susceptibility testing (VITEK 2. Results: The correlation between the two procedures has demonstrated a correlation of 98% for the strains identification; the correlation was of 100% for MRSA and ESBL recognition. Conclusions: The results showed that the use of the direct test for both Gram positive and Gram negative bacteria can give a complete report regarding the identification and susceptibility testing within 24 hours from when the sample becomes positive instead of 48h required using the conventional method.The procedure has demonstrated a high reliability both in terms of strains identification and of antibioticsusceptibility. We therefore can suggest, in the diagnostic of sepsis, the introduction of direct method in normal laboratory practice.

  19. Evaluation of the Verigene® Blood Culture Nucleic Acid test for rapid identification of gram positive pathogens from positive blood cultures

    Agnese Cellini


    Full Text Available Background. The rapid identification of the etiology and the evaluation of the antimicrobial susceptibility of the bacteria causing bacteremia is of outmost relevance to set up an adequate treatment of sepsis. In this study we evaluated the microarray based method, Verigene Gram-positive blood cultures (BC-GP nucleic acid test (Nanosphere Inc., Northbrook, IL, USA for the identification of Gram positive pathogens from positive blood cultures. The panel BC-GP is capable to identify 13 germs and 3 genes associated with antimicrobial resistance. Materials and Methods. In this study a total of 100 positive, non replicated and monomicrobic blood cultures have been evaluated. For testing on the Verigene platform using the BC-GP assay, 350 L of blood culture media from a positive the blood culture bottle.Results. A total of 100 positive blood cultures were tested by the Verigene BC-GP assay: out of these a total of 100 Gram-positive cocci were identified. The most frequent bacteria identified included staphylococci, streptococci and enterococci. Among staphylococci, Staphylococcus aureus accounted for 25% (15/60, with 38% of S. epidermidis 37% (23/60 and 37% (22/60 other CoNS. All the S. aureus isolates were correctly identified by BC-GP whereas in 2/45 cases (4% BC-GP misidentified CoNS. In the case of enterococci 7/10 were E. faecalis and 3 E. faecium, all of these were correctly identified.Conclusions. The overall agreement with the results obtained by standard procedure is quite elevated (88% and as a consequence the BC-GP panel could be used as a rapid diagnostic tool to give a faster response in the case of bacteremia associated with sepsis.

  20. Direct Testing of Blood Cultures for Detection of Streptococcal Antigens

    Wetkowski, Maryellen A.; Peterson, Ellena M.; de la Maza, Luis M.


    A direct, rapid, and simple method for the detection of streptococcal antigens of Lancefield groups A, B, C, D, and G from blood cultures was developed by using a coagglutination test. Fifty-five clinical specimens and 117 simulated blood cultures containing gram-positive cocci were tested. Out of 6,261 clinical blood cultures screened, 55 cultures from 53 patients were positive, with organisms resembling streptococci, by Gram stain. Of these cultures, 78% (43 of 55) were pure cultures of str...

  1. Large-scale clinical comparison of the lysis-centrifugation and radiometric systems for blood culture

    The Isolator 10 lysis-centrifugation blood culture system (E. I. du Pont de Nemours and Co., Inc., Wilmington, Del.) was compared with the BACTEC radiometric method (Johnston Laboratories, Inc., Towson, Md.) with 6B and 7D broth media for the recovery of bacteria and yeasts. From 11,000 blood cultures, 1,174 clinically significant organisms were isolated. The Isolator system recovered significantly more total organisms, members of the family Enterobacteriaceae, Staphylococcus spp., and yeasts. The BACTEC system recovered significantly more Pseudomonas spp., Streptococcus spp., and anaerobes. Of the Isolator colony counts, 87% measured less than 11 CFU/ml of blood. Organisms, on an average, were detected the same day from each of the two culture systems. Only 13 of the 975 BACTEC isolates (0.01%) were recovered by subculture of growth-index-negative bottles, and 12 of the 13 were detected in another broth blood culture taken within 24 h. Contaminants were recovered from 4.8% of the Isolator 10 and 2.3% of the BACTEC cultures

  2. Staphylococcus species and their Methicillin-Resistance in 7424 Blood Cultures for Suspected Bloodstream Infections

    Ariana ALMAŞ


    Full Text Available Objectives: The aim of this study was to evaluate the distribution of Staphylococcus species in bloodstream infections and to assess their susceptibility to methicillin. Material and Methods: Between January 1st 2008 - December 31st 2010, 7424 blood culture sets were submitted to the Laboratory Department of the Hospital for Clinical Infectious Diseases in Cluj-Napoca, Romania. The blood cultures were performed using BacT/Alert until January 2010 and BacT/Alert 3D automated system (bioMérieux after that date. The blood culture bottles were incubated at 37°C in a continuously monitoring system for up to 7 days. The strain identifications were performed by conventional methods, ApiStaph galleries and Vitek 2 Compact system. Susceptibility to methicillin was determined by disk diffusion method with cefoxitin disk and by using Vitek 2 Compact system. Results: From the total number of performed blood cultures, 568 were positive with Staphylococcus species. From 168 bacteriemic episodes 103 were with Staphylococcus aureus. Among 65 coagulase-negative staphylococci isolates, Staphylococcus epidermidis was the most frequently isolated species (34, followed by Staphylococcus hominis (15, Staphylococcus haemolyticus (8, Staphylococcus saprophyticus (3, Staphylococcus cohnii (1, Staphylococcus auricularis (1, and 3 strains that were not identified at species level. Methicillin resistance was encountered in 53.40% of Staphylococcus aureus strains and in 80% of coagulase-negative staphylococci. Conclusions: An important percentage of blood cultures were contaminated with Staphylococcus species. The main species identified in true bacteriemia cases were Staphylococcus aureus and Staphylococcus epidermidis. The percentage of methicillin-resistance, proved to be high not only for coagulase-negative staphylococci but also for Staphylococcus aureus.

  3. Bloodstream Infection in Neutropenic Cancer Patients Related to Short-Term Nontunnelled Catheters Determined by Quantitative Blood Cultures, Differential Time to Positivity, and Molecular Epidemiological Typing with Pulsed-Field Gel Electrophoresis

    Seifert, Harald; Cornely, Oliver; Seggewiss, Kerstin; Decker, Mathias; Stefanik, Danuta; Wisplinghoff, Hilmar; Fätkenheuer, Gerd


    To determine the rate of catheter-related bloodstream infection (CRBSI) among cases of primary bloodstream infection (BSI) in febrile neutropenic cancer patients with short-term nontunnelled catheters, quantitative paired blood cultures (Isolator) from the central venous catheter (CVC) and peripheral vein were obtained between November 1999 and January 2001. Bactec blood culture bottles were obtained to determine the differential time to positivity (DTP). CRBSI was defined as a quantitative b...

  4. New method for rapid Susceptibility Testing on blood culture with HB&L system: preliminary data

    Vincenzo Rondinelli


    Full Text Available Blood culture, although represents the gold standard in detecting the ethiological agent of sepsis, is rather rarely required in relation to the real diagnostic importance. The result of this test depends in fact on many factors (sample volume, time of collection, accuracy, antibiotic therapy, contamination, number of drawings, drawing site, interpretation difficulties, etc. that are often considered by many clinicians so limited as to doubt about their actual value. The disadvantages are therefore represented by the lack of standardization but also by the low sensitivity and above all by the technical times too long for the clinical needs. Blood culture begins with the drawing of samples from the “septic” patient followed incubation of the bottles in automatic thermostated systems. In case of positive result (36 hours, the culture is Gram stained and streaked on solid media in order to obtain isolated colonies for the identification and the susceptibility testing (48 hours from positive result. The long time required for pathogen identification and susceptibility testing involves empirical broad spectrum antibiotic therapy that can promote the increase of bacterial resistance but also patient management costs. A clinically useful report should be available on short notice in order to guide the clinician to choose the most appropriate antibiotic. The microbiologist has therefore the hard work of reviewing the organization and the management of the procedures.We have therefore started to consider the possibility of treating the blood as an biological liquid in order to quickly determine the susceptibility of bacteria to antibiotics.

  5. Many Bottles for Many Flies: Managing Conflict over Indigenous Peoples’ Cultural Heritage in Western Australia

    David Ritter


    This article critically considers the legal regulation of Indigenous people's cultural heritage in Western Australia and its operation within the framework of Australia's federal system of government. The article also sets out the different ways in which Indigenous cultural heritage is conceptualised, including as a public good analogous to property of the crown, an incidental right arising from group native title and as the subject of private contract. The article explores the various notion...

  6. Many Bottles for Many Flies: Managing Conflict over Indigenous Peoples’ Cultural Heritage in Western Australia

    David Ritter


    Full Text Available This article critically considers the legal regulation of Indigenous people's cultural heritage in Western Australia and its operation within the framework of Australia's federal system of government. The article also sets out the different ways in which Indigenous cultural heritage is conceptualised, including as a public good analogous to property of the crown, an incidental right arising from group native title and as the subject of private contract. The article explores the various notions of 'Indigenous cultural heritage' that exist under Western Australian public law and the significant role of private contractual arrangements. Particular attention is devoted to the uneasy nexus between the laws of native title and heritage in Western Australia.

  7. Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture

    Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.


    Background Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4–90.1%) and 62.5% (95% CI 24.5–91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as

  8. Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system

    In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia. To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory

  9. Identification of Brucella by MALDI-TOF Mass Spectrometry. Fast and Reliable Identification from Agar Plates and Blood Cultures

    Ferreira, Laura; Vega Castaño, Silvia; Sánchez-Juanes, Fernando; González-Cabrero, Sandra; Menegotto, Fabiola; Orduña-Domingo, Antonio


    Background MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. Methodology/Principal Findings We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. Conclusions/Significance MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles. PMID:21151913

  10. Bottled Water and Fluoride

    ... Fluoridation Journal Articles for Community Water Fluoridation Bottled Water Recommend on Facebook Tweet Share Compartir Consumers drink ... questions about bottled water and fluoride. Does bottled water contain fluoride? Bottled water products may contain fluoride, ...

  11. Recovery of clinically important microorganisms from the BacT/Alert blood culture system does not require testing for seven days.

    Wilson, M L; Mirrett, S; Reller, L B; Weinstein, M P; Reimer, L G


    Recently, we published a comparison of the BacT/Alert blood culture system with the BACTEC 660/730 nonradiometric blood culture system using blood inocula of 5 ml per bottle. By reanalyzing data collected during that study, we found that, for true-positive isolates causing bacteremia or fungemia, 363 (97.6%) of 376 and 341 (97.7%) of 349 isolates were recovered by the end of day 5 of testing, and 364 (97.9%) of 376 and 343 (98.3%) of 349 isolates were recovered by the end of day 6 of testing for aerobic and anaerobic bottles, respectively. Most isolates recovered on days 6 (24 of 27) and 7 (20 of 25) of testing were either contaminants or indeterminate as a cause of sepsis. When used as recommended by the manufacturer, only six (1.3%) of 464 clinically important isolates recovered on test days 6-7 would have gone undetected had testing been limited to 5 days and four (0.9%) of 464 had testing been limited to 6 days. We conclude that BacT/Alert bottles can be tested for as few as 5 days and then discarded with minimal loss of true-positive isolates and maximal reduction of contaminants. PMID:8425375

  12. Shortened Time to Identify Staphylococcus Species from Blood Cultures and Methicillin Resistance Testing Using CHROMAgar

    Shingo Chihara


    Full Text Available The ability to rapidly differentiate coagulase-negative staphylococcus (CoNS from Staphylococcus aureus and to determine methicillin resistance is important as it affects the decision to treat empiric antibiotic selection. The objective of this study was to evaluate CHROMagar S. aureus and CHROMagar MRSA (Becton Dickinson for rapid identification of Staphylococcus spp. directly from blood cultures. Consecutive blood culture bottles (BacT Alert 3D SA and SN, bioMérieux growing gram-positive cocci in clusters were evaluated. An aliquot was plated onto CHROMagar MRSA (C-MRSA and CHROMagar S. aureus (C-SA plates, which were read at 12 to 16 hours. C-SA correctly identified 147/147 S. aureus (100% sensitivity; 2 CoNS were misidentified as S. aureus (98% specificity. C-MRSA correctly identified 74/77 MRSA (96% sensitivity. None of the MSSA isolates grew on C-MRSA (100% specificity. In conclusion, CHROMagar is a rapid and sensitive method to distinguish MRSA, MSSA, and coagulase-negative Staphylococcus and may decrease time of reporting positive results.

  13. Evaluation of Fastidious Anaerobe Broth as a blood culture medium.

    Ganguli, L. A.; Turton, L J; Tillotson, G S


    Three commercial blood culture media were compared with a freshly prepared cooked meat medium in tests to stimulate the recovery of small inocula of anaerobic and aerobic bacteria in routine blood cultures. The cooked meat medium gave the most reliable recovery and supported continued viability, whilst Fastidious Anaerobe Broth (LAB M) was a good alternative. Results with Southern Group thioglycollate and Difco Thiol were less satisfactory as delays in recovery and loss of viability occurred ...

  14. Rapid detection of Pseudomonas aeruginosa from positive blood cultures by quantitative PCR

    Cattoir Vincent


    Full Text Available Abstract Background Pseudomonas aeruginosa is responsible for numerous bloodstream infections associated with severe adverse outcomes in case of inappropriate initial antimicrobial therapy. The present study was aimed to develop a novel quantitative PCR (qPCR assay, using ecfX as the specific target gene, for the rapid and accurate identification of P. aeruginosa from positive blood cultures (BCs. Methods Over the period August 2008 to June 2009, 100 BC bottles positive for gram-negative bacilli were tested in order to evaluate performances of the qPCR technique with conventional methods as gold standard (i.e. culture and phenotypic identification. Results Thirty-three strains of P. aeruginosa, 53 strains of Enterobactericaeae, nine strains of Stenotrophomonas maltophilia and two other gram-negative species were isolated while 3 BCs were polymicrobial including one mixture containing P. aeruginosa. All P. aeruginosa clinical isolates were detected by qPCR except a single strain in mixed culture. Performances of the qPCR technique were: specificity, 100%; positive predictive value, 100%; negative predictive value, 98.5%; and sensitivity, 97%. Conclusions This reliable technique may offer a rapid (

  15. Direct MALDI-TOF Mass Spectrometry Assay of Blood Culture Broths for Rapid Identification of Candida Species Causing Bloodstream Infections: an Observational Study in Two Large Microbiology Laboratories

    Spanu, Teresa; Posteraro, Brunella; Fiori, Barbara; D'Inzeo, Tiziana; Campoli, Serena; Ruggeri, Alberto; Tumbarello, Mario; Canu, Giulia; Trecarichi, Enrico Maria; Parisi, Gabriella; Tronci, Mirella; Sanguinetti, Maurizio; Fadda, Giovanni


    We evaluated the reliability of the Bruker Daltonik's MALDI Biotyper system in species-level identification of yeasts directly from blood culture bottles. Identification results were concordant with those of the conventional culture-based method for 95.9% of Candida albicans (187/195) and 86.5% of non-albicans Candida species (128/148). Results were available in 30 min (median), suggesting that this approach is a reliable, time-saving tool for routine identification of Candida species causing...

  16. Rapid Detection of Coliforms Using Cell Culture Bottle%用细胞培养瓶法快速检测大肠菌群

    段学辉; 魏斌; 傅奇; 郭炳其; 贾奎艳


    研究了一种快速检测大肠菌群的细胞培养瓶剂法,即以国标法中的乳糖胆盐发酵培养基为基本营养成分,经浓缩、干燥后制成快速检测瓶剂,待检样品滴入细胞培养瓶内,培养24h,依据培养瓶盖上产气指示试纸颜色变化及瓶内溶液颜色变化,确定可疑阳性样品,并对其进行氧化酶实验,验证大肠菌群的存在.实验对细胞培养瓶法进行了大肠菌群的重复性及实际样品检测,并与国标法进行比较.结果显示:细胞培养瓶法与国标法所测结果符合率达96%,样品检测报告时间只需24h,比国标法减少48 h,2种方法所测结果在统计学上无显著性差异(P>0.05).细胞培养瓶法简单、快速、准确,利于基层监测机构和生产企业对食品中大肠菌群的快速检测.%A cell culture bottle method was developed for rapid detection of coliforms in food samples. In this study, lactose bile salt medium were concentrated and dried for bottle testing agent. According to the color change of the gas test strip and the solution in cell culture bottle, suspected positive samples could be determined, and then be verified the presence of coliforms in suspected samples through oxidize and microscopic examination. The results of repetitive tests and actual samples detection were compared with national standard method. A high agreement (96% ) was found between the cell culture bottle method and national standard method on 50 samples. The overall assay time of cell culture bottle method only needed 24 h, which reduced 48 h than that of national standard method. The statistical analysis showed that there werent significant difference (P >0. 05) between the examination results for these methods. As a simple,rapid and accurate detection method,cell culture bottle could meet the needs of the primary monitoring agency and manufacturers for rapid detection of coliforms in food.

  17. Nurses' competency in drawing blood cultures and educational intervention to reduce the contamination rate.

    Al-Hamad, Arif; Al-Ibrahim, Maha; Alhajhouj, Eman; Al-Alshaikh Jaffer, Waseelah; Altowaileb, Jaffar; Alfaraj, Hassan


    Compared with truly negative cultures, false positive blood cultures (BCs) not only increase laboratory work but also prolong the lengths of patient stays, which are likely to increase patient morbidity and costs. The present study aimed to evaluate the effectiveness of a hospital-wide educational intervention on BC contamination rates. Nurses performed all phlebotomies; therefore, educational workshops were offered to all nurses twice a week over a 3-month period. The workshops consisted of a questionnaire, PowerPoint presentation, video show, demonstration of the different materials used to collect BCs, and question session. Data from the questionnaires and laboratory culture results were compared between the 6-month pre- and post-intervention periods. Of the 503 eligible nurses, 216 (42.9%) attended the workshops. The survey identified areas for improvement, which included time of disinfectant application, volume of blood to be cultured, and disinfection of BC bottle tops. Of the 9903 BC sets that were drawn from 3649 patients during the study period, 676 (6.8%) were contaminated. The monthly BC contamination rates for the 6-month pre- and post-intervention periods were 8.1% and 5.2%, respectively, representing a 36% reduction (P=0.008). Only three wards had an acceptable contamination rate of ≤3% before the intervention, compared with eight wards after the intervention. While contamination of BCs can never be completely eliminated, there is evidence that adherence to best practice BC collection techniques can minimize BC contamination, which might be best achieved with a dedicated phlebotomy team. PMID:26166815

  18. Radiometric detection of yeasts in blood cultures of cancer patients

    During a 12-month period, 19,457 blood cultures were collected. Yeasts were isolated from 193 cultures derived from 76 cancer patients. Candida albicans or Candida tropicalis accounted for 79% of isolates. Of the three methods compared, the radiometric method required 2.9 days to become positive, blind subculture required 2.6 days, and Gram stains required 1 day. However, the radiometric method was clearly superior in detecting positive cultures, since 73% of all cultures were first detected radiometrically, 22% were detected by subculture, and only 5% were detected by Gram stain. Although 93% of the isolates were detected by aerobic culture, five (7%) isolates were obtained only from anaerobic cultures. Seven days of incubation appear to be sufficient for the radiometric detection of yeasts

  19. Liberty Bottle or Liability Bottle?

    Livingstone, Verity


    The number of mothers initiating breastfeeding has increased dramatically over the last decade but the percentage of mothers who terminate breastfeeding prematurely has remained constant. When mothers experience difficulty with breastfeeding, many physicians fail to diagnose and manage the problem effectively. Some physicians assume that mothers dislike breastfeeding and, in a misguided attempt to help, recommend the introduction of a bottle to solve the problem. They do not explain to the mo...

  20. Modified mouse peripheral blood lymphocyte culture system for cytogenetic analysis

    A detailed methodology is presented for culturing mouse peripheral blood lymphocytes isolated on density gradients and stimulated to divide using either phytohemagglutinin, concanavalin A, or lipopolysaccharide. The techniques described yield more than sufficient numbers of mitotic cells for analyzing sister chromatid exchange, chromosome, aberrations, and micronuclei following in vitro or in vivo exposure to chemicals or radiation

  1. Survival and function of phagocytes in blood culture media

    Fischer, T K; Prag, J; Kharazmi, A


    The survival and function of human phagocytes in sterile aerobic and anaerobic blood culture media were investigated using neutrophil morphology, white blood cell count in a haemoanalyser, flow cytometry, oxidative burst response, and bactericidal effect in Colorbact and Septi-Chek blood culture...... media and Bact/Alert. When comparing agitation to stationary incubation no difference in phagocytic activity was found. The methods showed the same trends demonstrating that the phagocytes' viability and activity were prolonged by oxygen and shortened by anaerobic conditions and sodium polyethanol...... sulfonate (SPS). Best preserved activity and viability were found in the aerobic media containing less than 0.5 g/l SPS, in which significant phagocyte oxidative burst and bactericidal activity were found up to 4 days after inoculation. Considering that the majority of bacteremias are due to aerobic or...

  2. Trypanosoma cruzi. Surface antigens of blood and culture forms

    The surface polypeptides of both cultured and blood forms of Trypanosoma cruzi were iodinated by the glucose oxidase-lactoperoxidase technique. Blood-form trypomastigotes (BFT) isolated form infected mice displayed a major 90,000-Mr component. In contrast, both epimastigotes and trypomastigotes obtained form acellular cultures expressed a smaller 75,000-Mr peptide. Both major surface components were presumably glycoproteins in terms of their binding to concanavalin A-Sepharose 4B. Within a 3-h period, both blood and culture forms synthesized their respective surface glycoproteins (90,000 Mr and 75,000 Mr, respectively in vitro. [/sub 35/S]methionine-labeled surface peptides were immunoprecipitated with immune sera of both human and murine origin. A panel of sera form patients with chronic Chagas' disease and hyperimmunized mice recognized similar surface peptides. These immunogens were the same components as the major iodinated species. The major BFT surface peptide was readily removed by trypsin treatment of the parasites, although the procedure did not affect the 75,000-Mr peptide from the culture forms. Two-dimensional polyacrylamide gel electrophoresis revealed that the 90,000-Mr peptide found on BFT was an acidic protein of isoelectric point (pI) 5.0, whereas, the 75,000-Mr peptide form culture-form trypomastigotes has a pI of 7.2. The 90,000-Mr component is thought to be responsible for the anti-phagocytic properties of the BFT

  3. [Refinement of presumptive antimicrobial therapy based on initial microbiological information on positive blood culture].

    Aoki, Yosuke


    Positive blood culture represents either true bacteremia or contaminants of the normal skin flora. The number of positive bottles, rapidity with which blood culture turns positive, and appropriate interpretation of Gram-stain findings usually assist physicians or technologists in deciding whether it reflects true-positive results or contamination. In the case of true bacteremia, two aspects of the Gram-stain findings, Gram-positive or negative, cocci or rod, are important initial findings that safely guide physicians to select appropriate antimicrobial agents. Gram-positive cocci in clusters strongly suggest Staphylococci, and "in-chains" indicates Streptococci or Enterococci. Although distinction between the latter two organisms is occasionally difficult, glycopeptide should be the first choice, especially in critically ill patients. Gram-positive rods, when first reported, also require the empiric administration of glycopeptides, and sometimes their false Gram-negative staining could result in errors of pathogen identification, resulting in the inappropriate choice of antibiotics. The detection of gas production by Gram-negative rods, which indicates Enterobacteriaceae, is helpful initial information to start cephalosporin antibiotics, whereas the absence of gas would suggest nonfirmentative rod bacteremia, for which the administration of anti-pseudomonal agents is strongly warranted. Gram-negative cocci, such as Moraxella or Acinetobacter sp., may initially be reported as Gram-positive, so empiric antimicrobial drugs should be carefully selected taking into account these pitfalls and patients' conditions, and the situation regarding the development of diseases (community-acquired vs. nosocomial). The rapid and appropriate treatment of bacteremia thus requires careful interepretation of Gram-stain findings as described above, and should always be integrated with pathognomonic features of individual patients. PMID:20560459

  4. Should blood cultures be performed in terminally Ill cancer patients?

    Nobuhiro Asai


    Full Text Available Background: No evidence-based guidelines or protocols to treat the infection-related symptoms in cancer patients with terminal stages have been established. Materials and Methods: We retrospectively analyzed all the patients with terminal stage cancer who died between April 2009 and March 2010. The patients' background, the prevalence of infection and clinical outcomes, pathogens isolated, antibiotics used, and whether blood cultures and some of examinations were performed or not were evaluated. Results: A total of 62 (44 males and 18 females patients were included in this study. The median age was 73 years (35-98 years. The most common cancer was that of the lung (n =59, 95.2%. A total of 32 patients were diagnosed with the following infections: Infection of respiratory tract in 27 (84.4%, of urinary tract in 4 (12.5%, and cholangitis in 1 (3.1%. Two cases (6.3% had pneumonia complicated with urinary tract infection. Blood cultures and antibiotic therapies were performed in 28 and 30 cases, respectively. Four (14.3% positive cultures were isolated from the blood obtained from 28 individual patients. As for clinical course, 3 (10% of them experienced improved symptoms after antibiotic therapy. Twenty-seven (90% patients were not confirmed as having any symptom improvement. Conclusions: Blood cultures and antibiotic therapy were limited, and might not be effective in terminally ill cancer patients with lung cancer. We suggest that administering an antibiotic therapy without performing a blood culture would be one of choices in those with respiratory tract infections if patients' life expectancy is short.

  5. Evaluation of PNA-FISH yeast traffic light for rapid identification of yeast directly from positive blood cultures and assessment of clinical impact.

    Stone, N R H; Gorton, R L; Barker, K; Ramnarain, P; Kibbler, C C


    The PNA-FISH Yeast Traffic Light assay was performed on 54 clinical isolates of yeasts inoculated into blood culture bottles. The assay showed high sensitivity (Candida albicans/C. parapsilosis, 100%; C. glabrata/C. krusei, 92.3%; C. tropicalis, 100%) and specificity (C. albicans/C. parapsilosis, 100%; C. glabrata/C. krusei, 94.8%; C. tropicalis, 100%). Case note review estimated a change in therapy in 29% of cases had the PNA-FISH result been available to the clinician. PMID:23390280

  6. Blood culture cross contamination associated with a radiometric analyzer

    During a 9-day period in August 1980 in a New Jersey hospital, three pairs of consecutively numbered blood cultures from different patients were identified as positive for the same organism, for each pair, both cultures were positive in the same atmosphere, both organisms had the same sensitivities, and the second of each pair grew at least 2 days after the first and was the only positive blood culture obtained from the patient. When the hospital laboratory discontinued use of its radiometric culture analyzer for 15 days, no more consecutive pairs of positive cultures occurred. Subsequent use of the machine for 9 days with a new power unit but the original circuit boards resulted in one more similar consecutive pair (Staphylococcus epidermidis). After replacement of the entire power unit, there were no further such pairs. Examination of the machine by the manufacturer revealed a defective circuit board which resulted in inadequate needle sterilization. Laboratories which utilize radiometric analyzers should be aware of the potential for cross contamination. Recognition of such events requires alert microbiologists and infection control practitioners and a record system in the bacteriology laboratory designed to identify such clusters

  7. Staphylococci with markers of antibiotic resistance collected from blood cultures

    Vittorio Focarelli


    Full Text Available Introduction: Blood culture is still the gold standard for the detection of the causative agent of sepsis. Especially in intensive care patients and those with vascular catheters, the most common organisms isolated are coagulase-negative staphylococci (CoNS and Staphylococcus aureus, both characterized by multidrug resistance. Purposes of our work are the study of the incidence of markers of resistance in staphylococci and evaluation of potential changes over the years. Materials and methods: In the period January 2008-June 2011 5239 blood cultures were analyzed.They were mainly obtained from the departments of Intensive Care, Cardiology, Hematology, General Medicine, Emergency Medicine, Infectious Diseases, Oncology, Pulmonology and Pediatric Hematoncology. The vials containing the blood were incubated in the BACTEC 9120 automated tool of Becton Dickinson and susceptibility testing performed with the Phoenix instrument of the same company. Results:Within a total of 5239 blood cultures, 3967 (75.7% were negative and 1272 (24.3% positive. Fungi were isolated in 6.2% (79 of the positive ones, Gram-negative bacteria in 24.6% (313 and Gram-positive bacteria in 69.2% (880. Within the latter, 187 (21.2% were not staphylococcal isolates, 693 (78.8% were stafiloccocci mainly represented by S. epidermidis, S. aureus, S. hominis, S. haemolyticus and S. saprophyticus. Of the 693 staphylococcal isolates, 436 (62.9% were b lactamase producers, and between them 336 (77.1% were methicillin resistant, while only 3 of 436 (0.69% were S. aureus resistant to vancomycin as well.The incidence of markers of resistance was very high, especially in patients in intensive care and cardiac surgery, who are usually subjected to combined antibiotic therapy. In the three years studied there were no statistically significant differences in the resistance of staphylococci. Conclusions: The data show an alarming high number of multi-resistant staphylococci, which is often a

  8. A change of culture: reducing blood culture contamination rates in an Emergency Department.

    Bentley, James; Thakore, Shobhan; Muir, L; Baird, Alastair; Lee, Jennifer


    Blood cultures are an important investigation to help tailor effective management for patients with severe sepsis. Frequent contaminated samples increase laboratory workload and can delay or cause incorrect changes to patient management. This can prolong patient hospitalisation, increase the risk of harm and increase cost to health boards. Current guidelines advocate a contamination rate of 2-3%. From January 2013 to November 2014 inclusive, the contamination rate was 4.74% in our Emergency Department, responsible for initial management and investigation of over 40 cases of sepsis per month. A Quality Improvement team was created to try to reduce contamination rates to the recommended target. An initial baseline survey of local staff showed good understanding of when to obtain a blood culture but there was variability in the methods and equipment used. A project was then conducted which focused on rationalising and standardising equipment and technique for blood culture sampling along with staff education to support this change. A simple department target of 30 days free from a contaminated blood culture was created which, if achieved, would ensure a contamination rate of less than 3%. This was supported by ongoing surveillance of contamination rates and investigation of contaminated sample cases. We were able to then identify high risk patients and factors which increased the chance of blood culture contamination. This allowed us to formulate solutions to help reduce the risks of contamination. Department achievements and learning points to help prevent further contamination were fed back positively to all staff. This project operated for 12-months and successfully reduced local contamination rates to 2.0%. PMID:27335646

  9. Bacterial culture of perfusion blood after open-heart surgery.

    Freeman, R; Hjersing, N


    The results of routine culture of 595 consecutive specimens of perfusion blood are presented. Ten per cent of the specimens yielded bacteria overall, but it was found that the isolation rate was increased to 17.7% when the prophylactic antibodies being given during the bypass were specifically neutralised. Coagulase-negative staphylococci and diphtheroids formed the majority of organisms isolated, but Gram negative bacilli or "coliform" type were also occasionally found. A comparison of the relative findings in patients receiving prophylactic flucloxacillin or cephradine showed that the isolation rates of coagulase-negative staphylococci and diphtheroids were lower in the group receiving flucloxacillin. The origin of the bacteria isolated from perfusion blood remains uncertain but speciation of coagulase-negative staphylococci from perfusion blood and similar organisms isolated subsequently from catheter tips in the same patients revealed no evidence that the two sources of organisms were linked. Although organisms are easily and commonly found in perfusion blood, the relevance of this phenomenon to post-operative endocarditis is not clear. PMID:7008242

  10. Faster Identification of Pathogens in Positive Blood Cultures by Fluorescence In Situ Hybridization in Routine Practice

    Peters, Remco P. H.; Savelkoul, Paul H. M.; Simoons-Smit, Alberdina M.; Danner, Sven A.; Vandenbroucke-Grauls, Christina M. J. E.; van Agtmael, Michiel A.


    Rapid identification of microorganisms in blood cultures is required to optimize empirical treatment at an early stage. Fluorescence in situ hybridization (FISH) can reduce the time to identification of microorganisms in growth-positive blood cultures. In this study, we evaluated the performance, time to identification, and potential clinical benefits of FISH compared to those of conventional culture methods in routine practice. After Gram staining, blood culture fluids were simultaneously fu...


    A. Sobhani


    Full Text Available Bacteremia is an important infectious disease which may lead to death. Common bacteria and pattern of antibiotic resistance in different communities are different and understanding these differences is important. In the present study, relative frequency and pattern of drug resistance have been examined in bacteria isolated from blood cultures in Razi Hospital laboratory. The method of the study was descriptive. Data collection was carried out retrospectively. Total sample consisted of 311 positive blood cultures from 1999 to 2001. Variables under study were bacterial strains, antibiotics examined in antibiogram, microbial resistance, and patients' age and sex. The most common isolated bacteria were Salmonella typhi (22.2% and the least common ones were Citrobacter (1.6%. The highest antibiotic resistance was seen against amoxicillin (88.4%. The proportion of males to females was1: 1/1 and the most common age group was 15-44 (47.3%. Common bacteria and pattern of antibiotic resistance were different in some areas and this subject requires further studies in the future.

  12. Rapid Identification of Pathogens in Positive Blood Culture of Patients with Sepsis: Review and Meta-Analysis of the Performance of the Sepsityper Kit

    Morgenthaler, Nils G.; Kostrzewa, Markus


    Sepsis is one of the leading causes of deaths, and rapid identification (ID) of blood stream infection is mandatory to perform adequate antibiotic therapy. The advent of MALDI-TOF Mass Spectrometry for the rapid ID of pathogens was a major breakthrough in microbiology. Recently, this method was combined with extraction methods for pathogens directly from positive blood cultures. This review summarizes the results obtained so far with the commercial Sepsityper sample preparation kit, which is now approved for in vitro diagnostic use. Summarizing data from 21 reports, the Sepsityper kit allowed a reliable ID on the species level of 80% of 3320 positive blood culture bottles. Gram negative bacteria resulted consistently in higher ID rates (90%) compared to Gram positive bacteria (76%) or yeast (66%). No relevant misidentifications on the genus level were reported at a log(score)cut-off of 1.6. The Sepsityper kit is a simple and reproducible method which extends the MALDI-TOF technology to positive blood culture specimens and shortens the time to result by several hours or even days. In combination with antibiotic stewardship programs, this rapid ID allows a much faster optimization of antibiotic therapy in patients with sepsis compared to conventional workflows. PMID:26000017

  13. Rapid method for identification of group B streptococci in neonatal blood cultures.

    Holmes, R. L.; Harada, W A


    A rapid technique used for the identification of Streptococcus agalactiae, Lancefield group B, from the blood cultures of two neonatal infants is reported. The method utilized the Phadebact Streptococcus Test System (Pharmacia Diagnostics, Piscataway, N.J.) and the supernatant from 13- and 14-h blood cultures. Additional studies with simulated neonatal blood cultures revealed that this method was reproducible. Additional studies also revealed that some non-specific agglutination did occur, wh...

  14. 在静态和动态培养条件下杂交瘤细胞的生长和代谢%Growth and Metabolism of Hybridoma Cells Cultured in Flask and Spinner Bottles

    张立; 沈红; 张元兴


    The growth and metabolism of WuT3 hybridoma cells were very different when cells were cultured in flask and spinner bottles. In the flask bottles, a longer culture duration and the higher cell density were met, whereas in the spinner bottles the more vigorous cell metabolism took place, resulting in that either the specific consumption rates of glucose and amino acids or the specific production rates of lactate, ammonia and alanine were higher.%通过对WuT3杂交瘤细胞在静态和动态培养条件下的比较,发现细胞生长和代谢有很大不同。在静态培养条件下,细胞培养周期较长,细胞密度较高;但在动态培养条件下,细胞代谢更加旺盛,葡萄糖、氨基酸等营养物质的消耗加快,乳酸、氨、丙氨酸等代谢产物的比生成速率较大。

  15. Evaluation of an automated rapid diagnostic assay for detection of Gram-negative bacteria and their drug-resistance genes in positive blood cultures.

    Masayoshi Tojo

    Full Text Available We evaluated the performance of the Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN; Nanosphere, Northbrook, IL, USA, an automated multiplex assay for rapid identification of positive blood cultures caused by 9 Gram-negative bacteria (GNB and for detection of 9 genes associated with β-lactam resistance. The BC-GN assay can be performed directly from positive blood cultures with 5 minutes of hands-on and 2 hours of run time per sample. A total of 397 GNB positive blood cultures were analyzed using the BC-GN assay. Of the 397 samples, 295 were simulated samples prepared by inoculating GNB into blood culture bottles, and the remaining were clinical samples from 102 patients with positive blood cultures. Aliquots of the positive blood cultures were tested by the BC-GN assay. The results of bacterial identification between the BC-GN assay and standard laboratory methods were as follows: Acinetobacter spp. (39 isolates for the BC-GN assay/39 for the standard methods, Citrobacter spp. (7/7, Escherichia coli (87/87, Klebsiella oxytoca (13/13, and Proteus spp. (11/11; Enterobacter spp. (29/30; Klebsiella pneumoniae (62/72; Pseudomonas aeruginosa (124/125; and Serratia marcescens (18/21; respectively. From the 102 clinical samples, 104 bacterial species were identified with the BC-GN assay, whereas 110 were identified with the standard methods. The BC-GN assay also detected all β-lactam resistance genes tested (233 genes, including 54 bla(CTX-M, 119 bla(IMP, 8 bla(KPC, 16 bla(NDM, 24 bla(OXA-23, 1 bla(OXA-24/40, 1 bla(OXA-48, 4 bla(OXA-58, and 6 blaVIM. The data shows that the BC-GN assay provides rapid detection of GNB and β-lactam resistance genes in positive blood cultures and has the potential to contributing to optimal patient management by earlier detection of major antimicrobial resistance genes.

  16. Comparison of BD Bactec Plus Blood Culture Media to VersaTREK Redox Blood Culture Media for Detection of Bacterial Pathogens in Simulated Adult Blood Cultures Containing Therapeutic Concentrations of Antibiotics▿

    Miller, Nancy S.; Rogan, Dagmar; Orr, Beverley L.; Whitney, Dana


    Antibiotic neutralization in blood culture media from two automated systems was evaluated by measuring the recovery of organisms and times to detection in simulated cultures. Overall, BD Bactec Plus media (Bactec FX system) outperformed TREK 80 ml Redox media (VersaTREK system), although results suggest a relative rather than an absolute increased rate of recovery for the Bactec media.

  17. Bloodstream Infections Caused by Pseudomonas spp.: How To Detect Carbapenemase Producers Directly from Blood Cultures

    Dortet, Laurent; Boulanger, Anne; Poirel , Laurent; Nordmann, Patrice


    The Carba NP test has been evaluated to detect carbapenemase-producing Pseudomonas spp. directly from blood cultures. This rapid and cost-effective test permits an early identification of carbapenemase-producing Pseudomonas spp. directly from blood cultures with excellent sensitivity and specificity. Results may be useful in particular for guiding the first-line therapy and epidemiological purposes.

  18. 血培养实验室污染菌群分布与阳性报警时间的判断%Laboratory contamination bacterial distribution of blood culture and judgment of positive alarm time

    答嵘; 吴友伟; 王伟; 雷金娥


    . Results This research contained 17 941 bottles of blood culture ,including 7 193 sets of negative and 837 sets of posi‐tive ,the complete set rate was 89 .5% .The positive rate of blood culture was 10 .4% ,of which the contamination rate was 1 .6% .The main contaminated bacteria were coagulase negative staphylococcus ,and average alarm time of con‐taminated bacteria in aerobic and anaerobic bottles was 1 .34 ,1 .29 d ,respectively .The pathogenic bacteria and oppor‐tunistic pathogenic bacteria were common in the culture results of two bottles with the inconsistent culture results . Conclusion Blood culture contaminating bacteria are mainly skin resident bacteria .To reduce the contamination rate of blood culture ,it is vital to strictly execute the standard blood sampling procedure ,and emphasize the sterilization of blood‐collecting environment in hospitalized patients .

  19. Homocysteine induces production of monocyte chemoattractant protein-1 and interleukin-8 in cultured human whole blood

    Xiao-kun ZENG; Daniel G REMICK; Xian WANG


    AIM: To investigate whether increased plasma L-homocysteine (Hcy) level could promote monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in cultured whole blood. METHODS: Human whole blood or different type of peripheral blood cells from health volunteers were incubated with Hcy and/or the inhibitors. MCP- 1 and IL-8 level were measured by ELISA assay. RESULTS: Hcy 10-1000 μmol/L induced production of MCP-1 and IL-8 in cultured human whole blood (P<0.05). The major cellular source of these chemokines comed from monocytes.Meanwhile,Hcy also promoted the upregulation of MPO level even at the 10 μmol/L in the cultured whole blood.secretion in cultured human whole blood, especially in monocytes via oxidative stress mechanism.

  20. The effects of Saccharum officinarium (sugar cane) molasses on cytokine secretion by human blood cultures.

    Rahiman, Farzana; Pool, Edmund John


    This study investigated the effects of sugar cane molasses on the immune system, using cytokines as biomarkers. Whole blood cultures, stimulated in vitro with endotoxin or PHA, were incubated with various concentrations of molasses. No cell death occurred in whole blood cultures incubated with molasses samples. The addition of molasses (800 microg/mL) to unstimulated whole blood cultures resulted in increased levels of the biomarker of inflammation, Interleukin-6 (P Molasses addition (800 microg/mL) to unstimulated whole blood cultures has no effect on the cell mediated immunity biomarker, Interferon gamma secretion. Molasses has no effect on Interleukin-6, Interleukin-10 and Interferon gamma secretion in stimulated whole blood cultures. Immunostimulation by molasses requires further investigation as it may have potential health impacts. PMID:20391026

  1. A Comparative Study of Blood Culture Sampling from Umbilical Catheter Line versus Peripheral Site

    Abdolkarim Hamedi


    Full Text Available Neonatal sepsis is an important cause of death and morbidity in newborns and is diagnosed by isolation of organism in blood culture. In several reports,reliablity of blood cultures were done from umbi lical catheters,have been demonstrated. The objective of the present study was to determine,wether an inde welling umbilical catheter, could be an alternative site for blood culture. In a prospective study over 6 months during 2006,141 paired blood cultures from 134 infant,were done simultaneously from peripheral site and umbilical catheter (mostly U. V. C,during the first four days of life. Majority of these infants were preterm and admitted to NICU for special care. these infants had indwelling umbilical line and had indication of sepsis workup. A total of 141 pairs of blood cultures were obtained from 134 infants. In 16 infants blood culture pairs were positive for one organism in both peripheral vein and umbilical site. 71. 6% of total cultures (n=11pairs were negative in boths site. A total of 22 pairs were positive in one site only,with 5 positive from peripheral vein only and the other 17 from umblical site. Two pairs were positve in boths site with two different organism. In over all 16 infant (11%of blood were considered to be contaminated. Contamination rate were 2. 4% and 9. 2% for peripheral and umbilical catheter site. Contamination rate increased after 48 hours of age in umbilical catheter. The result showed that after 2 days contamination rate for blood culture taken from catheter line increased and specifity decreased. We recommended that blood culture via umblical catheter in first 2 days in sick neonates with indwelling catheter can be a alternate site of blood culture sampelling.

  2. Two Cases of Bacterial Pneumonia in Bottle-nosed Dolphins (Tursiops Gillii at the Seoul Zoo, Korea

    Kyung-Yeon Eo and Oh-Deog Kwon


    Full Text Available Two bacterial pneumonia cases of bottle-nosed dolphins (Tursiops gillii were found in Seoul Zoo. Firstly, severe fibrous pleuropneumonia was revealed in a 14-year-old male bottle-nosed dolphin on its necropsy. The clinical signs included anorexia, listing posture, and vertical submerging after the initial signs of low activity and avoiding trainers. White blood cell count increased up to 31.80 × 103/µl. Despite aggressive antibiotics therapy of intramuscular injection and nebulization it died two months later. At early time of illness Staphylococcus aureus and Proteus mirabilis were isolated. Lately Pseudomonas aeruginosa was isolated from the exhaled air culture and direct sample of lung parenchyma on necropsy. Secondly, a 6-year-old male bottle-nosed dolphin with anorexia and tendency to avoid trainers was completely recovered after prompt treatment with antibiotics and NSAIDs for six days prior to diagnosis. The diagnosis was based on history, clinical signs, blood cell count, and mucous culture from blowhole. The count of white blood cell dramatically decreased from 25.36 × 103/µl to 7.28 × 103/ µl after four days of antibiotics therapy. Staphylococcus aureus was isolated from the culture.

  3. Standardization and Assessment of Cell Culture Media Quantities in Roller Poly Ethylene Terephthalate Bottles Employed in the Industrial Rabies Viral Vaccine Production

    S. Jagannathan; S. Chaansha; K. Rajesh; T. Santhiya; C Charles; K.N. Venkataramana


    Vero cells are utilized for production of rabies vaccine. This study deals with the optimize quantity media require for the rabies vaccine production in the smooth roller surface. The rabies virus (Pasteur vaccine strain) is infected to monolayer of the various experimented bottles. To analyze the optimal quantity of media for the production of rabies viral harvest during the process of Vero cell derived rabies vaccine. The trials are started from 200 to 400 mL (PTARV-1, PTARV-2, PTARV-...

  4. Improved Amplification of Microbial DNA from Blood Cultures by Removal of the PCR Inhibitor Sodium Polyanetholesulfonate

    Fredricks, David N.; Relman, David A


    Molecular methods are increasingly used to identify microbes in clinical samples. A common technical problem with PCR is failed amplification due to the presence of PCR inhibitors. Initial attempts at amplification of the bacterial 16S rRNA gene from inoculated blood culture media failed for this reason. The inhibitor persisted, despite numerous attempts to purify the DNA, and was identified as sodium polyanetholesulfonate (SPS), a common additive to blood culture media. Like DNA, SPS is a hi...

  5. Time to positivity in blood cultures of adults with Streptococcus pneumoniae bacteremia

    Ansorena Luis; Garrido Jose; Rodríguez-Lera María; Peralta Galo; Roiz María


    Abstract Background previous studies have established that bacterial blood concentration is related with clinical outcome. Time to positivity of blood cultures (TTP) has relationship with bacterial blood concentration and could be related with prognosis. As there is scarce information about the usefulness of TTP, we study the relationship of TTP with clinical parameters in patients with Streptococcus pneumoniae bacteremia. Methods TTP of all cases of Streptococcus pneumoniae bacteremia, detec...

  6. Blood-group-related carbohydrates are expressed in organotypic cultures of human skin and oral mucosa

    Grøn, B; Andersson, A; Dabelsteen, Erik


    the function of cell-surface carbohydrates, we established organotypic cultures of skin and buccal mucosa. In these cultures, keratinocytes are grown at the air-liquid interface on a supporting matrix consisting of homologous fibroblasts embedded in a collagen type I gel. We examined the expression of blood-group...

  7. Time to positivity in blood cultures of adults with Streptococcus pneumoniae bacteremia

    Ansorena Luis


    Full Text Available Abstract Background previous studies have established that bacterial blood concentration is related with clinical outcome. Time to positivity of blood cultures (TTP has relationship with bacterial blood concentration and could be related with prognosis. As there is scarce information about the usefulness of TTP, we study the relationship of TTP with clinical parameters in patients with Streptococcus pneumoniae bacteremia. Methods TTP of all cases of Streptococcus pneumoniae bacteremia, detected between January 1995 and December 2004 using the BacT/Alert automated blood culture system in a teaching community hospital was analyzed. When multiple cultures were positive only the shortest TTP was selected for the analysis. Results in the study period 105 patients with Streptococcus pneumoniae bacteremia were detected. Median TTP was 14.1 hours (range 1.2 h to 127 h. Immunosuppressed patients (n = 5, patients with confusion (n = 19, severe sepsis or shock at the time of blood culture extraction (n = 12, those with a diagnosis of meningitis (n = 7 and those admitted to the ICU (n = 14 had lower TTP. Patients with TTP in the first quartile were more frequently hospitalized, admitted to the ICU, had meningitis, a non-pneumonic origin of the bacteremia, and a higher number of positive blood cultures than patients with TTP in the fourth quartile. None of the patients with TTP in the 90th decile had any of these factors associated with shorter TTP, and eight out of ten patients with TTP in the 10th decile had at least one of these factors. The number of positive blood cultures had an inverse correlation with TTP, suggesting a relationship of TTP with bacterial blood concentration. Conclusion Our data support the relationship of TTP with several clinical parameters in patients with Streptococcus pneumoniae bacteremia, and its potential usefulness as a surrogate marker of outcome.

  8. Evaluation of osteogenic cell culture and osteogenic/peripheral blood mononuclear human cell co-culture on modified titanium surfaces

    This study aimed to determine the effect of a bioactive ceramic coating on titanium in the nanothickness range on human osteogenic cells, peripheral blood mononuclear cells (PBMC) and on osteogenic cells co-cultured with PBMC without exogenous stimuli. Cell viability, proliferation, adhesion, cytokine release (IL1β, TGFβ1, IL10 and IL17) and intracellular stain for osteopontin and alkaline phosphatase were assessed. Morphologic evaluation showed smaller and less spread cell aspects in co-culture relative to osteogenic cell culture. Cell viability, proliferation and adhesion kinetics were differently influenced by surface texture/chemistry in culture versus co-culture. Cytokine release was also influenced by the interaction between mononuclear and osteogenic cells (mediators released by mononuclear cells acted on osteogenic cells and vice versa). In general, ‘multi-cell type’ interactions played a more remarkable role than the surface roughness or chemistry utilized on the in vitro cellular events related to initial stages of bone formation. (paper)

  9. Recovery of Histoplasma capsulatum from blood in a commercial radiometric Mycobacterium medium.

    Merz, W G; Kodsy, S; Merz, C S


    We report the recovery of Histoplasma capsulatum from blood specimens cultured for Mycobacterium sp. in BACTEC 13A radiometric medium. H. capsulatum was recovered from six of eight blood specimens submitted for mycobacterial cultures from five human immunodeficiency virus-positive individuals. Initial positive metabolic signals occurred at a mean of 11 days, but no organisms were detected with acid-fast stains. The bottles remained positive, and after an additional incubation (mean, 8 days), ...

  10. Homocysteine induces production of monocyte chemoattractant protein-1 and interleukin-8 in cultured human whole blood

    Xiao-kunZENG; DanielGREMICK; XianWANG


    AIM: To investigate whether increased plasma L-homocysteine (Hcy) level could promote monocyte chemoattract antprotein-1 (MCP-1) and interleukin-8 (IL-8) in cultured whole blood. METHODS: Human whole blood or differenttype of peripheral blood cells from health volunteers were incubated with Hcy and/or the inhibitors. MCP-1 and IL-8 level were measured by ELISA assay. RESULTS: Hcy 10-1000 μmol/L induced production of MCP-1and IL-8 in cultured human whole blood (P<0.05). The major cellular source of these chemokines comed from monocytes. Meanwhile,Hcy also promoted the upregulation of MPO level even at the 10 μmol/L in the cultured whole blood.The intracellular ROS, particular the OH radicals, play extremely important role in the Hcy-induced MCP-1 and IL-8 production. CONCLUSION: Increased Hcy level in plasma (hyperhomocysteinemia) induced MCP-1 and IL-8secretion in cultured human whole blood, especially in monocytes via oxidative stress mechanism,

  11. Philadelphia chromosome detection in chronic myeloid leukemia: Utility of phytohemagglutinin-stimulated peripheral blood culture

    Man Updesh Singh Sachdeva


    Full Text Available Background: The conventional cytogenetic approach to demonstrate Philadelphia (Ph chromosome at times does not yield enough number of metaphases or are of suboptimal quality. Further, the rapid molecular tests have completely pushed this simple technique into disrepute. Aims: This study aimed to evaluate usefulness of phytohemagglutinin (PHA-stimulated peripheral blood culture for detection of Ph chromosome in chronic myeloid leukemia (CML patients. Materials and Methods: Fifty-six patients, including 11 newly diagnosed cases of CML and 45 patients of CML on imatinib therapy showing the presence of Ph chromosome in unstimulated samples, were included in the study. Cytogenetic analysis was done on unstimulated samples, i.e. bone marrow aspirate, 24- and 48-h peripheral blood culture, and compared with PHA-stimulated 72-h peripheral blood culture. Results: The preparations from PHA-stimulated peripheral blood culture samples in all 56 patients yielded high number of good-quality metaphases. All the 11 (100% newly diagnosed patients and 39/45 (87% of the patients on imatinib therapy showed the presence of Ph chromosome in PHA-stimulated samples. Addition of PHA-stimulated 72-h peripheral blood culture preparation can be of use for increasing the diagnostic yield in cases of CML with suboptimal results on conventional cytogenetics from bone marrow aspirate sample.

  12. Blood Culture Contamination in Children’s Medical Center of Tehran from April to July 2004

    F Shhcheraghi


    Full Text Available Background: Blood culture is the criterion standard for identifying children with bacteremia. However, elevated false-positive rates are common and are associated with substantial health care costs. The aims of this prospective study were to: 1 determine the rate of blood culture contamination 2 determine variety and frequency of contaminant bacteria 3 compare the duration of hospital stay and antibiotic administration in patients with true bacteremia vs those have false positive blood culture. Materials and Methods: Cross-sectional study conducted April through July 2004 among patients aged 14 years or younger who were admitted at Doctor Garib Children Medical Center of Tehran and had a blood culture obtained as part of their care. Bacterial isolates were identified to species level and medical records were reviewed in all cases with a positive blood culture. A number of clinical and laboratory criteria were used to deciding whether a blood isolate is a pathogen or a contaminant. These include the identify of the micro-organism itself, clinical features such as fever and leukocytosis; the proportion of blood culture sets positive as a function of the number of sets obtained and to have an indwelling vascular catheter or prosthetic device. Results: During the study period, 2877 sets of blood culture were evaluated and the rates of positive blood cultures associated with significant bacteremia and contamination were 1.04% and 5.4% respectively. Among the positive blood cultures, over the 84% of isolates were due to contamination and only 15.95% of isolated strains associated with true infection. The frequency of isolated bacteria with respect to true infection and contamination are as following: S. Aureus (infect: 9.0%, contam: 0.0%, S. Epidemidis (infec: 0.0%, contam: 13.3%, Micrococcus sp. (infec: 0.0%, contam: 4.3%, pseudomonas and related species other than P. aeruginosa (infec: 2.1%, contam: 60.6%, viridans group of streptococci (infec: 1

  13. Direct identification and susceptibility testing of gram-negative bacilli from BACTEC bottles by use of the MS-2 system with updated bacterial identification software.

    Dipersio, J R; Ficorilli, S M; Varga, F J


    The Abbott MS-2 system (Abbott Laboratories, Diagnostic Division, Irving, Tex.), equipped with updated bacterial identification software (version 03.02), was used to perform both direct identification and susceptibility tests on gram-negative bacilli from positive BACTEC blood culture bottles. Ninety-eight of 101 Enterobacteriaceae strains, one strain of Acinetobacter calcoaceticus, and two strains of Pseudomonas aeruginosa were correctly identified by following a direct inoculation procedure...


    Kaur Satbir


    Full Text Available Bottle gourd is one of the excellent fruits gifted by nature to human beings having composition of all the essential constituents that are required for good health and quality human life. Lagenaria siceraria (Cucurbitaceae, popularly known as bottle gourd, lauki or ghiya, is a climbing plant, which bears hard-shelled and bottle-shaped gourds as fruits. It forms an excellent diet being rich in vitamins, iron and minerals. The fruit is reported to contain the triterepenoide cucurbitacins B, D, G, H, two sterols viz., fucosterol and campesterol, aerpene byonolic acid (an allergic compound, flavone-C glycosides (a ribosome inactivating protein and lagenin. Extract of the ghiya seeds show antibiotic activity. The fruit juice is helpful in constipation, premature graying hair, urinary disorders and insomnia. Lauki has the highest content of choline among all the vegetables known to man till date, which serves as the precursor of neurotransmitter acetylcholine, which in turn is crucial for retaining and enhancing memory. Furthermore, Lagenaria siceraria is a vegetable useful in the management of many diseases like cardiac disorders, hepatic diseases and ulcer. Bottle gourd juice helps to regulate blood pressure of hypertensive patients, because of its high potassium content. It helps in losing weight quickly, because of its high dietary fiber and low fat and cholesterol content. In the light of above facts, the authors have made a humble attempt to compile an up-to-date review article on Lagenaria siceraria covering its phytochemistry, pharmacological actions and folk medicinal uses.

  15. Commodification of nature : social construction of bottled spring water


    This thesis aims to investigate the social processes that are involved in constructing a commodity and a consumer need in a commercial society. In Norway bottled spring water is a symbol of nature while representing the power of marketing and consumer culture. The study explores how spring water has been transformed into a commodity without loosing its symbolic value. Ringnes launched Imsdal in 1994, and is regarded as the first successful brand within bottled spring water in this country. I ...

  16. The Study of Blood Culture for Prevalent Bacteria and Antibiotic Resistance on Hospitalized Patients

    H Alaodolei


    Full Text Available Background: Bacteremia means invasion of bacteria to coronary- arthery system. One third of these cases lead to septicemia and in 40-50% cases, it causes patient’s death. Therefore information about resistance and prevalent of bacteria isolated from blood culture is important for deciding about suitable therapeutic management. Methods: This retrospective study was done on all positive blood cultures for typing and detecting of antibiotic resistance during 2001- 2005. Data was analyzed by statistical procedure. Results: In 252 (4.35% of studied blood cultures, the most prevalent bacteries were Staph. epidermidis (35.2% and E. Coli (18.5%. The greatest and the least resistance antibiotics were βLactam (75.2% and glycopeptide (7.8% groups, respectively. Conclusion: With regard to antibiotic resistance increased during these years, awaring of the last changes about it in every therapeutic center is necessary.

  17. Mortality and prognostic factors of patients who have blood cultures performed in the emergency department

    Lindvig, Katrine P; Nielsen, Stig Lønberg; Henriksen, Daniel P; Jensen, Thøger G; Kolmos, Hans Jørn; Pedersen, Court; Vinholt, Pernille J; Lassen, Annmarie T


    ratio (HR) 4.6 (95% CI 3.6-6.0)], at least two organ failure [HR 3.6 (2.9-4.5)], bacteraemia [HR 1.4 (1.1-1.8)], Charlson Comorbidity Index of at least 2 h [HR 1.7 (1.3-2.0)], SIRS [HR 1.5 (1.2-1.7)], a history of alcohol dependency [HR 1.7 (1.3-2.3)] and late drawing of blood cultures 24-48 h after...... failure, bacteraemia, Charlson Comorbidity Index of at least 2, SIRS, a history of alcohol dependency and late drawing of blood cultures....

  18. Comparative usefulness of inflammatory markers to indicate bacterial infection-analyzed according to blood culture results and related clinical factors.

    Nishikawa, Hirokazu; Shirano, Michinori; Kasamatsu, Yu; Morimura, Ayumi; Iida, Ko; Kishi, Tomomi; Goto, Tetsushi; Okamoto, Saki; Ehara, Eiji


    To assess relationships of inflammatory markers and 2 related clinical factors with blood culture results, we retrospectively investigated inpatients' blood culture and blood chemistry findings that were recorded from January to December 2014 using electronic medical records and analyzed the data of 852 subjects (426 culture-positive and 426 culture-negative). Results suggested that the risk of positive blood culture statistically increased as inflammatory marker levels and the number of related factors increased. Concerning the effectiveness of inflammatory markers, when the outcome definition was also changed for C-reactive protein (CRP), the odds ratio had a similar value, whereas when the outcome definition of blood culture positivity was used for procalcitonin (PCT), the greatest effectiveness of that was detected. Therefore, the current results suggest that PCT is more useful than CRP as an auxiliary indication of bacterial infection. PMID:26525643

  19. Measurement of microbial DNA polymerase activity enables detection and growth monitoring of microbes from clinical blood cultures.

    Daniel R Zweitzig

    Full Text Available Surveillance of bloodstream infections (BSI is a high priority within the hospital setting. Broth-based blood cultures are the current gold standard for detecting BSI, however they can require lengthy incubation periods prior to detection of positive samples. We set out to demonstrate the feasibility of using enzymatic template generation and amplification (ETGA-mediated measurement of DNA polymerase activity to detect microbes from clinical blood cultures. In addition to routine-collected hospital blood cultures, one parallel aerobic blood culture was collected and immediately refrigerated until being transported for ETGA analysis. After refrigeration holding and transport, parallel-collected cultures were placed into a BACTEC incubator and ETGA time-course analysis was performed. Of the 308 clinical blood cultures received, 22 were BACTEC positive, and thus were initially selected for ETGA time course analysis. The ETGA assay detected microbial growth in all 22 parallel-positive blood cultures in less time than a BACTEC incubator and also yielded genomic DNA for qPCR-based organism identification. In summary, feasibility of detecting microbes from clinical blood culture samples using the ETGA blood culture assay was demonstrated. Additional studies are being considered towards development of clinically beneficial versions of this methodology.

  20. Holographic optical bottle beams

    Alpmann, Christina; Rose, Patrick; Denz, Cornelia


    We present a convolution approach for the generation of optical bottle beams that combines established techniques of holographic optical trapping with hollow intensity distributions in order to manipulate absorbing particles. The versatility of our method is demonstrated by the simultaneous stable trapping of multiple particles at defined positions. Furthermore, the presented phase shaping technique allows for the dynamic manipulation of absorbing particles along arbitrary paths.

  1. Clonal population of flucytosine-resistant Candida tropicalis from blood cultures, Paris, France

    Desnos-Ollivier, Marie; Bretagne, Stéphane; Bernède, Claire; Robert, Vincent; Raoux, Dorothée; Chachaty, Elisabeth; Forget, Elisabeth; Lacroix, Claire; Dromer, Françoise


    Candida tropicalis is a diploid ascomycetes yeast responsible for 4%-24% of candidemia. Resistance to flucytosine is rarely described for this species but was observed for 45 (35%) of 130 C. tropicalis isolates recovered from blood cultures in the Paris area in a 4-year survey. The aims of this stud

  2. Clinical manifestation and laboratory findings in positive blood culture in neonatal septicemia

    Gholamreza Khademi


    Full Text Available Background/objective: Neonatal septicemia is one of the major causes of mortality in newborns. The aim of this study is to evaluate the clinical manifestations and laboratory findings in positive blood culture in neonatal septicemia. Methods: In this retrospective study, we allocated 100 records positive blood culture of neonates suffering from septicemia. A questionnaire was completed for each patient consisting the age at admission, gender, weight at birth, admission time, type of delivery, pre- or post-term delivery and the clinical symptoms. Types of organism causing sepsis, and their resistance to antibiotics were evaluated and method for empirical treatment was recommended. Results: Respiratory distress, cyanosis and lethargy were more common in the patients. The antibiogram showed Ampicillin resistance in 86% and Gentamycin resistance in 66% of studied records. Also, 36% cases of positive blood culture with gram-negative and 64% with gram-positive bacteria were observed. The most common bacteria in blood cultures were negative-coagulase Staphylococcus (%35, Staphylococcus Aureus (%24, Klebsiella (%18, respectively. Other bacteria were Enterobacter, Escherichia coli and Enterococcus (%5, Acinetobacter (%3, Pseudomonas aeruginosa and Negative-Gram Bacilli (%2 and Ceratia (%1. The most common effective antibiotics against bacterial growth in Antibiograms were Vancomycin, Cephalosporin, Amikacin, Co-trimoxazole and Gentamycin. Conclusion: Since the most common bacteria in neonatal septicemia cases were negative-coagulase Staphylococcus, Staphylococcus Aureus, Klebsiella, the pediatricians must select the regiments that cover gram-negative bacteria for empirical antibiotic treatments.

  3. Antimicrobial Susceptibility and Microorganisms Isolated from Blood Cultures of Hospitalized Patients in Intensive Care Units

    Emine Küçükateş


    Full Text Available Aim: The aim of this study was to evaluate microorganism growth in blood cultures of hospitalized patients in our intensive care units and to determine appropriate antimicrobial agents for treatment. Methods: We retrospectively investigated the blood cultures obtained from the patients hospitalized in the Coronary and Surgical Intensive Care Units at the Institute of Cardiology, İstanbul University, between July 2013 and December 2014. All microorganisms were identified using the conventional methods. Results: A total of 1034 blood cultures were obtained from 324 patients. Microbial growth was detected in 174 (16.8% blood cultures of 68 patients. Among all microbial growth, 113 (58.55% were gram-positive bacteria, 69 (35.75% were gram-negative rods and 11 (5.7% were fungi. Staphylococcus aureus was the most frequent microorganism (48; 24.87%, followed by coagulasenegative Staphylococci (35; 18,13%, Enterococcus spp. (30; 15,54%, Stenotrophomonas maltophilia, Escherichia coli, and Pseudomonas spp. 60.4% of Staphylococcus aureus were methicillin-resistant and 65.7% of coagulase-negative Staphylococci were also methicillin-resistant. All Staphylococci and Enterococci were not resistant to vancomycin, teicoplanin and tigecycline. All the gram-negative rods were susceptible to colistin and tigecycline, followed by imipenem (71.6% and meropenem (70.7%. Conclusions: We assume that infection control measures must be increased due to high antibiotic resistance and besides, antibiotic policies should be improved.

  4. Western Culture in Japanese Film: Kurosawa's "Throne of Blood" and "Ran."

    Kane, Peter E.

    Akira Kurosawa, the most popular Asian film maker with audiences in the United States, has found in William Shakespeare's plays themes and plots that resonate within Japanese culture. While the translations of "Macbeth" into "Throne of Blood" and "King Lear" into "Ran" are quite direct and literal with only minor changes in plot and emphasis, in…

  5. Recent Progress in the Diagnosis of Pathogenic Candida Species in Blood Culture.

    Phoompoung, Pakpoom; Chayakulkeeree, Methee


    Candidemia has become an emerging invasive fungal disease. Prompt treatment with appropriate antifungal agent is crucial to reduce the mortality of candidemia. The conventional blood culture method, which is considered the gold standard for candidemia diagnosis, has a low sensitivity and is time-consuming to perform. Recently, several novel advanced diagnostic methods that have a higher sensitivity and a shorter turnaround time than the conventional blood culture method have been developed for the early detection of Candida in blood samples or in blood culture broth. Most of these newer methods were developed using various molecular techniques, such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, peptide nucleic acid fluorescence in situ hybridization, and a number of DNA-based techniques including in-house and commercial polymerase chain reactions. In this article, we review and summarize the novel molecular methods that have been recently used for the detection and identification of Candida organisms in blood specimens. PMID:27003437

  6. Ginseng, a Potentially Therapeutic Drug in γ-Irradiated Whole Blood Culture

    Ginseng is commonly used as herbal medicines with wide range of beneficial effects. It is also approved as effective agent against radiation hazards, through its immunomodulating role in irradiated experimental animals. This experiment aimed to assess cytogenetic and biochemical changes of ginseng at a working dose (100µg/ ml) in suppressing radiation effects of human peripheral blood. The treatment times were 6, 48 and 72h after γ-irradiation at dose of 4 Gy (the last two treatment periods were done through blood culture). Triple blood cultures for each blood sample were set up. Cytogenetic investigations were evaluated using chromosome aberration (CA) analysis and cytokinesis-block micronucleus assay (CBMN). The levels of malondialdhyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO) were estimated in blood plasma and in 48 and 72h blood cultures. In addition, immune function response was assessed by estimation of the levels of (Immunoglobulin G) IgG and IgM for the same treatment periods. Ionizing irradiation induced significant decrease of SOD activity. While, CA (dicentric, ring, breaks and polyploidy), micronucleus (MN), MDA level and NO concentration were significantly increased. For IgG and IgM, there is temporary over-production of them after 6h of irradiation, and then their levels decreased in the periods 48 and 72h within the cultures. Ginseng post-irradiation treatment exhibits increase in the level of IgG and IgM production, and improvement in NO concentration. In addition, there is reduction in the levels of lipid peroxidation (LPO), CA and MN frequencies. Results concluded that ginseng has antimutagenic effect and benefit against oxidative stress involved by irradiation

  7. Factors Associated with Blood Culture Contamination in the Emergency Department: Critical Illness, End-Stage Renal Disease, and Old Age.

    Chih-Jan Chang

    Full Text Available Blood culture contamination in emergency departments (ED that experience a high volume of patients has negative impacts on optimal patient care. It is therefore important to identify risk factors associated with blood culture contamination in EDs.A prospectively observational study in a university-affiliated hospital were conducted between August 2011 and December 2012. Positive monomicrobial and negative blood cultures drawn from adult patients in the ED were analyzed to evaluate the possible risk factors for contamination. A total of 1,148 positive monomicrobial cases, 391 contamination cases, and 13,689 cases of negative blood culture were identified. Compared to patients with negative blood cultures, patients in triage levels 1 and 2 (Incidence Rate Ratio, IRR = 2.24, patients with end-stage renal disease (ESRD (IRR = 2.05, and older patients (IRR: 1.02 per year were more likely to be associated with ED blood culture contamination.Critical patients (triage levels 1 and 2, ESRD patients, and older patients were more commonly associated with blood culture contamination in the ED. Further studies to evaluate whether the characteristics of skin commensals contribute to blood culture contamination is warranted, especially in hospitals populated with high-risk patients.

  8. Ease-of-use protocol for the rapid detection of third-generation cephalosporin resistance in Enterobacteriaceae isolated from blood cultures using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

    Foschi, C; Compri, M; Smirnova, V; Denicolò, A; Nardini, P; Tamburini, M V; Lombardo, D; Landini, M P; Ambretti, S


    An ease-of-use protocol for the identification of resistance against third-generation cephalosporins in Enterobacteriaceae isolated from blood culture bottles was evaluated using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. A cefotaxime hydrolysis assay from chocolate agar subcultures using antibiotic discs and without inoculum standardization was developed for routine work flow, with minimal hands-on time. This assay showed good performance in distinguishing between cefotaxime-susceptible and cefotaxime-resistant strains, with excellent results for Escherichia coli (sensitivity 94.7%, specificity 100%). However, cefotaxime resistance was not detected reliably in Enterobacteriaceae expressing AmpC genes or carbapenemase-producing Klebsiella pneumoniae. PMID:27105753

  9. Do Big Bottles Kickstart Infant Weight Issues?

    ... Do Big Bottles Kickstart Infant Weight Issues? Smaller baby bottles ... 2016 (HealthDay News) -- Feeding babies formula from a big bottle might put them at higher risk for ...

  10. The Utility of Blood Culture Fluid for the Molecular Diagnosis of Leptospira: A Prospective Evaluation.

    Dittrich, Sabine; Rudgard, William E; Woods, Kate L; Silisouk, Joy; Phuklia, Weerawat; Davong, Viengmon; Vongsouvath, Manivanh; Phommasone, Koukeo; Rattanavong, Sayaphet; Knappik, Michael; Craig, Scott B; Weier, Steven L; Tulsiani, Suhella M; Dance, David A B; Newton, Paul N


    Leptospirosis is an important zoonosis worldwide, with infections occurring after exposure to contaminated water. Despite being a global problem, laboratory diagnosis remains difficult with culture results taking up to 3 months, serology being retrospective by nature, and polymerase chain reaction showing limited sensitivity. Leptospira have been shown to survive and multiply in blood culture media, and we hypothesized that extracting DNA from incubated blood culture fluid (BCF), followed by quantitative real-time polymerase chain reaction (qPCR) could improve the accuracy and speed of leptospira diagnosis. We assessed this retrospectively, using preincubated BCF ofLeptospiraspp. positive (N= 109) and negative (N= 63) febrile patients in Vientiane, Lao PDR. The final method showed promising sensitivities of 66% (95% confidence interval [CI]: 55-76) and 59% (95% CI: 49-68) compared with direct or direct and indirect testing combined, as the respective reference standards (specificities > 95%). Despite these promising diagnostic parameters, a subsequent prospective evaluation in a Lao hospital population (N= 352) showed that the sensitivity was very low (∼30%) compared with qPCR on venous blood samples. The disappointingly low sensitivity does suggest that venous blood samples are preferable for the clinical microbiology laboratory, although BCF might be an alternative if leptospirosis is only suspected postadmission after antibiotics have been used. PMID:26880775

  11. Association of different types of milk feeding with blood culture positive neonatal sepsis

    To ascertain and compare microbial growth pattern in blood culture of septic neonates who were either totally breast or formula fed. Study Design: Cross sectional study. Place and Duration of Study: The Children's Hospital Lahore, Pakistan from Feb 2012 to Dec 2012. Methodology: All clinically septic neonates, who were either exclusively breast fed or formula fed, were enrolled in the study. They were divided into two groups and studied for the type of organisms grown on blood culture. Group-A were breast fed and group-B were formula fed. Neonates who were blood culture negative or had growth of multiple organisms or had incomplete data or who died / left against medical advice before completing the required data or babies receiving milk feeding from multiple sources or no feeding at all were excluded. BACTEC technique was used for obtaining bacterial growth. SPSS version 19 was used for statistical analysis. Results: A total of 380 clinically septic neonates were enrolled. Each group consisted of 190 subjects. Incidence of culture positive sepsis in breast fed and in formula fed was 6.7% and 15.7% respectively (p-value = 0.0001). Overall, gram-negative organisms constituted the majority (16.1%). Thirty seven percent cultures grew coagulase negative Staphylococcus (CoNS) followed by Klebsiella spp (23.4%). In group A, gram-negative and gram-positive organisms were equally distributed whilst in group-B, gram-negative organisms were three times more frequent than gram-positive organisms. Predominant pattern of organisms was also different in the two groups. In group-A, CoNS was predominant while in group-B, Klebsiella spp. was most frequent. Conclusion: Culture positive sepsis is more than two times greater in formula fed babies and is caused predominantly by gram-negative organisms whilst in breast fed babies, CoNS is the commonest organism. (author)

  12. Introduction of Bottle Gourd DNA into Watermelon by Soaking Embryo for Resistance to Wilt Disease


    In order to utilize wilt disease resistance of bottle gourd, total DNA of bottle gourd was introduced into watermelon through the method of soaking embryo. The DNA-introduced variant offsprings were cultured in contaminated soil to elect the wilt disease resistance for more than 3 generations. 2 high- resistant and 2 middle-resistant watermelon materials were obtained.

  13. Continuous and semi-continuous cell culture for production of blood clotting factors.

    Desai, Sunil G


    Recombinant clotting factors are important biotherapeutics that Pfizer has produced and marketed for over fifteen years. Owing to the complexity of the structure and function of these blood factors, it can be challenging to achieve the required product quality and manufacturing productivity. The article highlights the semi-continuous and continuous cell culture processes employed by Pfizer for the production of BeneFIX and ReFacto AF. The benefits of such processes, the challenges of maintaining an aseptic production culture for extended periods, and batch definition are discussed in this article. PMID:25738489

  14. Beacon-based (bbFISH®) technology for rapid pathogens identification in blood cultures

    Sakarikou, Christina; Parisato, Martina; Cascio, Giuliana Lo; Fontana, Carla


    Background Diagnosis and treatment of bloodstream infections (BSI) are often hampered by the delay in obtaining the final results of blood cultures. Rapid identification of pathogens involved in BSI is of great importance in order to improve survival of septic patients. Beacon-based fluorescent in situ hybridization (hemoFISH® Gram positive and hemoFISH® Gram negative test kits, miacom diagnostics GmbH Düsseldorf, Germany) accelerates the identification of most frequent bacterial pathogens of...

  15. Human whole-blood culture system for ex vivo characterization of designer-cell function.

    Schukur, Lina; Geering, Barbara; Fussenegger, Martin


    Encapsulated designer cells implanted into mice are currently used to validate the efficacy of therapeutic gene networks for the diagnosis and treatment of various human diseases in preclinical research. Because many human conditions cannot be adequately replicated by animal models, complementary and alternative procedures to test future treatment strategies are required. Here we describe a novel approach utilizing an ex vivo human whole-blood culture system to validate synthetic biology-inspired designer cell-based treatment strategies. The viability and functionality of transgenic mammalian designer cells co-cultured with primary human immune cells were characterized. We demonstrated that transgenic mammalian designer cells required adequate insulation from the human blood microenvironment to maintain viability and functionality. The biomaterial alginate-(poly-l-lysine)-alginate used to encapsulate the transgenic designer cells did neither affect the viability of primary granulocytes and lymphocytes nor the functionality of lymphocytes. Additionally, alginate-encapsulated transgenic designer cells remained responsive to the release of the pro-inflammatory cytokine tumor necrosis factor (TNF) from the whole-blood culture upon exposure to bacterial lipopolysaccharide (LPS). TNF diffused into the alginate capsules, bound to the specific TNF receptors on the transgenic designer cells' surface and triggered the expression of the reporter gene SEAP (human placental secreted alkaline phosphatase) that was rewired to the TNF-specific signaling cascade. Human whole-blood culture systems can therefore be considered as valuable complementary assays to animal models for the validation of synthetic circuits in genetically modified mammalian cells and may speed up preclinical research in a world of personalized medicine. PMID:26348251

  16. Bridging the gap between detection and confirmation of B. anthracis in blood cultures

    Hawkey, Suzanna


    The spore forming bacterium, Bacillus anthracis is the aetiological agent of anthrax. The 2001 US anthrax letter attacks and the 2009‐2010 outbreak of injectional anthrax in the UK highlighted the importance of early detection and confirmation of this agent, both for patient outcome and forensic investigations. A reliable and consistent method was used in this study to safely simulate blood cultures with B. anthracis and used to determine the time to positive detection. This was performed...

  17. Coagulase-negative staphylococci strains resistant to oxacillin isolated from neonatal blood cultures

    Valeria Cataneli Pereira


    Full Text Available Coagulase-negative staphylococci (CoNS are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.

  18. Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture

    P. Pranke


    Full Text Available Identification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO, FLT-3 ligand (FL and kit ligand (KL; or stem cell factor in different combinations: TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9% CD34+ cells, 2.6 ± 2.1% of which were CD38-negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25%. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29% of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype.

  19. Classification of Blood Culture Isolates Into Contaminants and Pathogens on the Basis of Clinical and Laboratory Data.

    Hossain, Belal; Weber, Martin W; Hamer, Davidson H; Hibberd, Patricia L; Ahmed, A S M Nawshad Uddin; Marzan, Mahfuza; Islam, Maksuda; Connor, Nicholas E; Islam, Mohammad Shahidul; Zaidi, Anita K; Baqui, Abdullah H; Bhutta, Zulfiqar A; Qureshi, Shahida M; Rafiqullah, Iftekhar; McGee, Lesley; Saha, Samir K


    The multisite community-based study, Aetiology of Neonatal Infection in South Asia (ANISA), uses blood culture as the gold standard for identifying the etiology of neonatal infection. Considering the importance of this age-old diagnostic tool and the risk of contamination, ANISA has employed rigorous measures to prevent contamination at all stages of blood collection, processing and culture. Because contamination may still occur, an independent expert group evaluates the routinely collected clinical and laboratory data to determine whether a blood culture isolate is a contaminant or a true pathogen. This article describes the methodology used by ANISA to determine whether a blood culture isolate is likely to be a true pathogen or a contaminant in neonatal sepsis. PMID:27070065

  20. Ocular injuries from flying bottle caps.

    Fonseka, C


    Three cases of serious eye injury are described from flying metal caps of carbonated drink bottles. The injuries occurred while attempting to open the bottle in an unconventional and dangerous way. Though injuries from flying bottle caps have been described before, they have occurred when the bottle exploded. This is the first report of eye injuries caused by bottle caps while opening and are similar to the injuries caused by champagne corks. PMID:8143337

  1. Effect of live yeast culture Saccharomyces cerevisiae on milk production and some blood parameters

    Judit Peter Szucs


    Full Text Available The aim of this study was to investigate the effect of live yeast culture (Saccharomyces cerevisiae Sc 47 on milk yield, milk composition and some blood parameters of dairy cows during their early lactation on farm conditions. The live yeast culture was given in the diet of heifers and cows (5 g day-1 solid Actisaf for 14 days before calving and exclusively for the treated cows 12 g day-1 dissolved in 500 ml of water, during 14 days after calving. The experiment took until 100th day of lactation on farm conditions. Yeast culture supplementation was the most effective for the performance of primiparous cows: It was advantageous for blod plasma parameters: decreased the beta-hydroxy butyrate (BHB content and free fatty acids (FFA which indicated the protection of the animals against ketosis or other metabolic disorders. Increased the daily milk production and the lactose /glucose content of the milk. The live yeast culture increased the lactose content of the milk and decreased the somatic cell count of multiparous cows. The listed parameters were not significant (P<0.05 compare to the results of positive control groups. The applied live yeast culture supplementation did not significant affect for other performance of the cows.

  2. Comparison of histopathological analysis, culture and polymerase chain reaction assays to detect mucormycosis in biopsy and blood specimens

    Parisa Badiee; Amir Arastefar; Hadis Jafarian


    Background and Objectives The aim of this study was to compare direct microscopic examination with culture and PCR for the diagnosis of Mucorales infection in blood and tissue specimens. Material and Methods Blood samples and tissue specimens were obtained from 28 patients (total 58 samples) with suspected invasive fungal infection and cultured on proper media. Direct smear of tissue samples was done with potassium hydroxide, hematoxylin and eosin, and methenamine silver staining. DNA extract...

  3. A recommendation to perform a blood culture before the administration of intravenous antibiotics increased the detection of Staphylococcus aureus bacteremia

    Jogenfors, A.; Stark, L.; Svefors, J.; Löfgren, S; Malmvall, B.-E.; Matussek, A.


    In 2004, the Surviving Sepsis Campaign was launched to increase awareness and improve the outcome of severe sepsis. Accordingly, in Jönköping County, Sweden, a strong recommendation to perform a blood culture before the start of intravenous antibiotic treatment was introduced in 2007. Moreover, a reminder was included in the laboratory report to consult an infectious disease specialist when Staphylococcus aureus was isolated from a blood culture. Retrospectively, patients with at least one bl...

  4. Rapid identification of pneumococci, enterococci, beta-haemolytic streptococci and S. aureus from positive blood cultures enabling early reports

    Larsson, Marie C; Karlsson, Ewa; Woksepp, Hanna; Frolander, Kerstin; Mårtensson, Agneta; Rashed, Foad; Annika, Wistedt; Schön, Thomas; Serrander, Lena


    BACKGROUND: The aim of this study was to evaluate diagnostic tests in order to introduce a diagnostic strategy to identify the most common gram-positive bacteria (pneumococci, enterococci, β-haemolytic streptococci and S. aureus) found in blood cultures within 6 hours after signalling growth. METHODS: The tube coagulase test was optimized and several latex agglutination tests were compared and evaluated before a validation period of 11 months was performed on consecutive positive blood cultur...

  5. Beers Straight from the Bottle

    Murphy, James Peter


    Consuming beer straight from the bottle a trend initiated by the US is now an accepted practice throughout Irish bars but that does not necessarily mean it is a good thing from either a hygiene or taste viewpoint.

  6. Evaluation of a direct blood culture disk diffusion antimicrobial susceptibility test.

    Doern, G V; Scott, D R; Rashad, A L; Kim, K S


    A total of 556 unique blood culture isolates of nonfastidious aerobic and facultatively anaerobic bacteria were examined by direct and standardized disk susceptibility test methods (4,234 antibiotic-organism comparisons). When discrepancies which could be accounted for by the variability inherent in disk diffusion susceptibility tests were excluded, the direct method demonstrated 96.8% overall agreement with the standardized method. A total of 1.6% minor, 1.5% major, and 0.1% very major discrepancies were noted. PMID:7325634

  7. Staphylococcus lugdunensis endocarditis--the hidden peril of coagulase-negative staphylococcus in blood cultures.

    Seenivasan, M H; Yu, V L


    Reported here is a successfully treated case of native mitral valve endocarditis caused by Staphylococcus lugdunensis and a review of 47 similar cases reported in the English literature. In the literature review, perineal skin flora appeared to be the source of the organism in patients with endocarditis. Staphylococcus lugdunensis is generally susceptible in vitro to beta-lactam agents. If speciation is not performed, these bacteria might be mistaken for Staphylococcus epidermidis, a relatively avirulent bacterium that is a common contaminant of cultures. Prompt speciation can lead to earlier recognition of endocarditis and possibly enable earlier surgical intervention with improved outcome for this high-mortality infection. Multiple positive blood cultures yielding coagulase-negative staphylococci should be identified to the species level; endocarditis or another intravascular source of infection should be sought. PMID:12845551

  8. Survival of Campylobacter jejuni in mineral bottled water according to difference in mineral content: Application of the Weibull model

    Guillou, Sandrine; Leguérinel, Ivan; Garrec, N.; Renard, M. A.; Cappelier, Jean-Michel; Federighi, Michel


    International audience The aim of the study was to examine the hypothesis proposed by Evans et al. [2003. Hazards of healthy living: bottled water and salad vegetables as risk factors for Campylobacter infection. Emerg. Infect. Dis. 9(10), 1219–1225] that mineral bottled water accidentally contaminated by Campylobacter jejuni would represent a risk factor for Campylobacter infection. Culturability of C. jejuni cells inoculated in low- and high-mineral bottled water during storage at 4 1C i...

  9. Importance of blood cultures from peripheral veins in pediatric patients with cancer and a central venous line

    Handrup, Mette Møller; Møller, Jens Kjølseth; Rutkjaer, Cecilie; Schrøder, Henrik


    When an infection is suspected in a child with cancer and a central venous line (CVL), cultures are often only obtained from the CVL and not from a peripheral vein (PV). This study was undertaken to evaluate the importance of concomitant blood cultures from the CVL and a PV....

  10. The Colorful Chemical Bottle Experiment Kit: From School Laboratory To Public Demonstration

    Limpanuparb, Taweetham


    The blue bottle experiment was first introduced to the chemical education literature as a simple demonstration on kinetics. Its original formulation contains only glucose, NaOH and small amount of methylene blue. The solution turns blue when shaken and fades to colorless upon standing. This bluing/de-bluing cycle may be repeated and may be compared to blood colors in animal's respiratory cycle. Inspired by the blue bottle experiment, the colorful chemical bottle experiment kit was commercially developed in 2006. The kit is a versatile pedagogical tool, not only for physical chemistry but also for analytical, biological and organic chemistry. It also helps teaching concepts in scientific method and laboratory safety. This manuscript contains four parts, brief review on literature relating to the blue bottle experiment, description of the colorful chemical bottle experiment kit, pedagogical discussion of the experiments and preliminary evaluation from students.

  11. Evaluating the role of low-speed centrifugation towards transfecting human peripheral blood mononuclear cell culture.

    Majumdar, M; Ratho, R; Chawla, Y; Singh, M P


    The conventional method of transfection of suspension cells by chemical has proven to be very difficult. We present a new transfection protocol, wherein, low-speed centrifugation of cell culture plates immediately after adding the lipid: DNA complex significantly enhances the transfection efficiency. Peripheral blood mononuclear cells (PBMCs) were transfected with BLOCK-iT™ Fluorescent Oligo (scrambled siRNA) and lipofectamine complex using conventional and low-speed centrifugation modified transfection protocols. The efficiency of transfection was determined using flowcytometer and cell viability was checked using MTT assay. Incorporation of low-speed centrifugation significantly enhances the transfection efficiency of BLOCK-iT™ in the suspension culture of PBMCs as compared to conventional transfection method (99.8% vs 28.3%; P saving and novel application without compromising the cell viability in the context of recently popular RNA interference in suspension cultures of PBMCs. This undemanding modification might be applicable to a wide variety of cell lines and solve crucial problem of researchers working with RNA interference in suspension cultures. PMID:24713904

  12. Evaluating the role of low-speed centrifugation towards transfecting human peripheral blood mononuclear cell culture

    M Majumdar


    Full Text Available The conventional method of transfection of suspension cells by chemical has proven to be very difficult. We present a new transfection protocol, wherein, low-speed centrifugation of cell culture plates immediately after adding the lipid: DNA complex significantly enhances the transfection efficiency. Peripheral blood mononuclear cells (PBMCs were transfected with BLOCK-iT™ Fluorescent Oligo (scrambled siRNA and lipofectamine complex using conventional and low-speed centrifugation modified transfection protocols. The efficiency of transfection was determined using flowcytometer and cell viability was checked using MTT assay. Incorporation of low-speed centrifugation significantly enhances the transfection efficiency of BLOCK-iT™ in the suspension culture of PBMCs as compared to conventional transfection method (99.8% vs 28.3%; P < 0.0001, even at a low concentration of 40 picomoles without affecting the cell viability. Centrifugation enhanced transfection (CET technique is simple, time-saving and novel application without compromising the cell viability in the context of recently popular RNA interference in suspension cultures of PBMCs. This undemanding modification might be applicable to a wide variety of cell lines and solve crucial problem of researchers working with RNA interference in suspension cultures.

  13. Combined education and skin antisepsis intervention for persistently high blood-culture contamination rates in neonatal intensive care.

    O'Connor, C; Philip, R K; Powell, J; Slevin, B; Quinn, C; Power, L; O'Connell, N H; Dunne, C P


    Contaminated blood cultures represent challenges regarding diagnosis, duration of hospitalization, antimicrobial use, pharmacy and laboratory costs. Facing problematic neonatal blood culture contamination (3.8%), we instigated a successful intervention combining skin antisepsis using sterile applicators with 2% chlorhexidine gluconate in 70% isopropanol prior to phlebotomy (replacing 70% isopropanol) and staff education. In the six months prior to intervention, 364 neonatal peripheral blood samples were collected. Fourteen (3.8%) were contaminated. In the post-intervention six months, 314 samples were collected. Three (0.96%) were contaminated, representing significant improvement (Fisher's exact test: P = 0.0259). No dermatological sequelae were observed. The improvement has been sustained. PMID:26944902

  14. Immunomodulatory effects of natural polysaccharides assessed in human whole blood culture and THP-1 cells show greater sensitivity of whole blood culture.

    Gill, Satbir Kaur; Islam, Nahidul; Shaw, Iain; Ribeiro, Andreia; Bradley, Benjamin; Brien, Timothy O'; Kilcoyne, Michelle; Ceredig, Rhodri; Joshi, Lokesh


    Immunomodulatory drugs are available to maintain immune homeostasis but some have undesirable side effects. Six oligo- and poly-saccharides were assessed for their pro- and anti-inflammatory responses in two in vitro model systems, the monocytic THP-1 cell line and human whole blood cultures (HWBC). The compounds were first characterised for their molecular mass and physical properties. Following incubation with lipopolysaccharide (LPS) or the compounds, cytokine and chemokine secretion was assayed in both models and intracellular TNF-α was measured by flow cytometry in HWBC cell sub-populations. LPS, inulin, galacturonan, heteroglycan and fucoidan demonstrated pro-inflammatory properties and intracellular TNF-α expression was increased in the monocytes of HWBC. Mannan and xyloglucan did not elicit any significant responses. Inulin induced maximum cytokine secretion and heteroglycan induced maximum chemokine secretion in HWBC. This study emphasises the potential of inulin and heteroglycan as potential immunomodulatory therapeutics and that HWBC had a greater and more varied response in comparison to THP-1 cells. PMID:27218669

  15. Comparative study of subculture, Gram staining and acridine orange staining for early detection of positive blood cultures.

    Mascart, G; Bertrand, F.; Mascart, P.


    In view of the importance of a rapid aetiological diagnosis in septicaemia, we compared the results of subculture, Gram staining and acridine orange staining in the detection of positive blood cultures. The study was based on 1013 blood cultures of which 138 were positive by culture. The three techniques were applied 12 h after the specimen was taken in 210 instances, at 24 h in 540 instances and after 48 h in 525. We were able to demonstrate the value of direct examination. Staining with acr...

  16. Study of Prevalence and Antimicrobial Susceptibility of Blood Culture Bacterial Isolates

    Ayobola, E. D.


    Full Text Available Bloodstream infections are associated with significant morbidity and mortality. Definitive diagnosis is by bacteriologic culture of blood samples to identify organisms and establish antibiotic susceptibility. Between July and September 2009, 249 blood samples collected from patients at the University of Benin Teaching Hospital were processed. Positive cultures which accounted for 48(19.3% of total samples screened, were purified and identified according to standard methods. Sensitivity of bacteria to different antibiotics was determined by Kirby-Bauer disk diffusion method. Microorganisms recovered were Staphylococcus aureus (14.6%, Providencia spp., Pseudomonas aeruginosa, Enterobacter spp., Klebsiella pneumoniae and Proteus mirabilis (12.5% respectively, Escherichia coli and Staphylococcus epidermidis (8.3% respectively and Citrobacter freundii (6.3% . The highest antibiotic activities against Gram positive isolates were observed for ofloxacin (90.9%, nitrofurantoin (81.8% and gentamicin (72.7%, while in Gram negative bacteria, ofloxacin (81.1% and nalidixic acid (45.9% were most effective. The possibility of drug resistance acquisition by bacteria makes continuous surveillance of antimicrobial susceptibility patterns of bacteria essential as this will enhance efforts to identify resistance and attempt to limit its spread.

  17. Bacteriologic Profile and Antibiogram of Blood Culture Isolates from a Children's Hospital in Kabul

    Objective: To identify the bacterial pathogens causing paediatric septicaemia in Kabul and to determine their antibiogram to improve empirical antibiotic therapy. Study Design: Cross-sectional study. Place and Duration of Study: Microbiology Laboratory of FMIC, Kabul, Afghanistan, from January 2010 to June 2012. Methodology: Blood cultures from suspected cases of sepsis were processed in BD (Becton Dickinson, USA) for culture BACTEC 9240 Blood Culture System. Positive growths were examined and isolates were identified by conventional biochemical tests. Bacteria were identified to the species level using various Analytical Profile Index (API) identification strips. Antibiotic susceptibility testing was performed by modified Kirby-Bauer disk diffusion method. Drug resistant strains were studied for extended spectrum beta lactamase (ESBL) production by combination disk method and for methicillin resistant Staphylococcus aureus (MRSA) by Cefoxitin disk diffusion method. Results: Out of a total 3360 blood cultures received from in-patients, 410 yielded monomicrobial growth; hence the frequency of positive blood culture was 12.2%. Out of a total 410 isolates, 212 (51.71%) were gram-negative bacilli and 184 (44.88%) were gram-positive cocci. In addition, 14 (3.41%) Candida species were also isolated. The frequently isolated species of gram-negative bacteria belonged to Enterobacteriaceae and included 66 Klebsiella (16.1%), 42 Enterobacter (10.2%), 35 Escherichia (E.) coli (8.5%) and 16 Serratia (3.9%) species. In addition, 21 (5.12%) Pseudomonas species were also isolated. Correspondingly, amongst gram-positive cocci, the most frequently isolated species were 108 coagulase-negative Staphylococci (26.34%) followed by 49 Staphylococcus aureus (11.95%) and 21 Streptococcus species (5.12%). Among gram-negative isolates, those that produced ESBL i.e., 110 out of 212 (51.9%) were found to be multidrug-resistant and showed high resistance to commonly used antibiotics namely

  18. Comparison of Blood Collected in Acid-Citrate-Dextrose and EDTA for Use in Human Immunodeficiency Virus Peripheral Blood Mononuclear Cell Cultures

    Fiscus, Susan A.; Chakraborty, Hrishikesh; Shepard, Robin; Goodman, Melissa


    Paired blood samples collected in acid-citrate-dextrose and EDTA were compared for human immunodeficiency virus (HIV) infectivity on the day of collection or after 1 day of storage at room temperature. No significant differences between the anticoagulants were observed. Culture positivity was significantly associated with HIV RNA viral loads for both anticoagulants.

  19. A Plastic Bottle in Rectosigmoid

    A. Derakhshanfar


    Full Text Available Introduction: Evaluation and treatment of foreign bodies in rectum involves careful history and physical examination. The cases of forced introduction of the objects most commonly are , sexual assault , self – introduced for anal eroticism and accidental insertion.Case Report: We describe a case of a patient with rectal impaction following self administration of a plastic bottle for anal sexual gratification. A 49 years old man was admitted in the emergency department with the history of self introduced a bottle into his rectum physical examination and abdominal X-Ray diagnosed the case as impacted foreign body in rectosigmoid. An attempt was made to deliver the bottle through the rectum but because of high lying big bottle in the sigmoid laporotomy was performed and the bottle was removed though a longitudinal incision on sigmoid colon.Conclusion: Retained rectosigmoid foreign bodies have been encountered more frequently and present a dilemma for management and rarely laporotomy for extraction of foreign bodies was performed.

  20. Induction of vascular endothelial phenotype and cellular proliferation from human cord blood stem cells cultured in simulated microgravity

    Chiu, Brian; Z-M Wan, Jim; Abley, Doris; Akabutu, John


    Recent studies have demonstrated that stem cells derived from adult hematopoietic tissues are capable of trans-differentiation into non-hematopoietic cells, and that the culture in microgravity ( μg) may modulate the proliferation and differentiation. We investigated the application of μg to human umbilical cord blood stem cells (CBSC) in the induction of vascular endothelial phenotype expression and cellular proliferation. CD34+ mononuclear cells were isolated from waste human umbilical cord blood samples and cultured in simulated μg for 14 days. The cells were seeded in rotary wall vessels (RWV) with or without microcarrier beads (MCB) and vascular endothelial growth factor was added during culture. Controls consisted of culture in 1 G. The cell cultures in RWV were examined by inverted microscopy. Cell counts, endothelial cell and leukocyte markers performed by flow-cytometry and FACS scan were assayed at days 1, 4, 7 and at the termination of the experiments. Culture in RWV revealed significantly increased cellular proliferation with three-dimensional (3D) tissue-like aggregates. At day 4, CD34+ cells cultured in RWV bioreactor without MCB developed vascular tubular assemblies and exhibited endothelial phenotypic markers. These data suggest that CD34+ human umbilical cord blood progenitors are capable of trans-differentiation into vascular endothelial cell phenotype and assemble into 3D tissue structures. Culture of CBSC in simulated μg may be potentially beneficial in the fields of stem cell biology and somatic cell therapy.

  1. Bottled Water Everywhere: Keeping it Safe

    ... regulations. These beverages are instead considered to be soft drinks. back to top It May Be Tap Water ... and Nutrient-Added Water Beverages Bottled Water/Carbonated Soft Drinks Guidance Documents & Regulatory Information How FDA regulates bottled ...

  2. 27 CFR 25.142 - Bottles.


    ... bottle, the nature of the product such as beer, ale, porter, stout, etc., and the place of production... 5100.31, when required by Part 7 of this chapter. (f) Short-fill bottles. A brewer may dispose...

  3. Lowest modes of a bottle

    Helmholtz's model predicts correctly the frequency of the lowest mode of a bottle. A simple generalization of Helmholtz's model correctly predicts this mode and also the next few modes: ''flute modes'': of flasks with long uniform necks but arbitrarily shaped bodies. Wine bottles have additional low-frequency ''cavity modes'' that require a further easy generalization. For a bottle with slowly varying cross section an additional generalization can be made that retains the one-dimensional (1-D) character of the previous models and gives results that are in good agreement with experiment for the lowest mode: the ''diametral mode'': of a hollow sphere. For higher modes of a sphere, the 1-D model is inadequate and must be discarded in favor of exact solutions of the 3-D wave equation

  4. 27 CFR 31.232 - Wine bottling.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Wine bottling. 31.232... OF THE TREASURY LIQUORS ALCOHOL BEVERAGE DEALERS Miscellaneous § 31.232 Wine bottling. Each person desiring to bottle, package, or repackage taxpaid wines must, before carrying on those operations,...

  5. Corrigendum - Effect of live yeast culture Saccharomyces cerevisiae on milk production and some blood parameters

    Judit Peter Szucs


    Full Text Available In the article Effect of live yeast culture Saccharomyces cerevisiae on milk production and some blood parameters, first published in 2013 in Scientific Papers: Animal Science and Biotechnologies, 46 (1, the first author neglected to ask any prior agreement and permission of Lesaffre Feed Additive firm the sponsor of the experiment and Bernhard Feix GmbH the leader of the experiment, for publishing the results, doing so she committed serious mistakes: Ethically: The first author published data for which she had no permission, and she did not indicate the name of the owners, neither did she refere to them. On the scientific basis: She was not able to evaluate properly the real effect of Saccharomices cerevisiae containing Actisaf additive since the introduced experiment was a basic large farm size /scale research work that could not be measured precisely without the planned and ongoing further experiments.   Acknowledgement The first author highly appreciates Lesaffre Feed Additive and Bernhard Feix GmbH firms for the possibility of taking part in the research work together with her students. She is most grateful for the generous promise of the leaders of both Lesaffre Feed Additive and Bernhard Feix GmbH for disregarding to take any legal steps. She gives thanks especially to the leader of the experiment Zoltan Pachinger for his wise and helpful contribution to this consensus. The first author apologises for this oversight.     This article corrects: Effect of live yeast culture Saccharomyces cerevisiae on milk production and some blood parameters, Vol. 46, Issue 1, p. 40-44. Article first published online: 30 May 2013

  6. Influence of normal blood serum on the uptake and organificition of iodine-131 by thyroid cell culture

    The effect of normal blood serum of cattle on the levels of uptake and organification of iodine-131 by the primary culture of thyroid cells of newborn porcine has been studied. The negative dose-dependent effect suggests the influence of biologically active substances of blood serum on the regulation of these processes both at the receptor and post receptor levels, but the mechanism of a such effect is yet unknown

  7. New method to differentiate human peripheral blood monocytes into insulin producing cells: Human hematosphere culture.

    Hur, Jin; Yang, Ji Min; Choi, Jae-Il; Yun, Ji-Yeon; Jang, Jae Hee; Kim, Joonoh; Kim, Ju-Young; Oh, Il-Young; Yoon, Chang-Hwan; Cho, Hyun-Jai; Park, Young-Bae; Kim, Hyo-Soo


    Strategy to differentiate stem cells into insulin producing cells (IPCs) in vitro has been a promising one to get cell source of β-cell replacement therapy for diabetes. It has been suggested that islets and neurons share features and nestin-positive cells could differentiate into IPCs. We have recently developed a three-dimensional culture system using human peripheral blood cells named as blood-born hematosphere (BBHS). Here we showed that most of BBHS were composed of nestin-positive cells. Under the four-stage differentiation protocol for IPCs, we plated nestin-positive BBHS onto fibronectin-coated dish. These cells form islet-like clusters and most of them expressed insulin. Pancreatic specific genes were turned on, such as transcription factors (Pdx-1, Ngn3 and Nkx6.1), genes related to endocrine function (Glut-2 and PC2) or β cell function (Kir6.2, SUR1). Furthermore islet differentiation was confirmed by dithizone (DTZ) staining to detect zinc ion which binds insulin protein within the cells. Finally, IPCs derived from BBHS showed capability to secrete insulin in response to glucose stimulation. Taken together, our novel protocol successfully induced islet-like human insulin producing cells out of BBHS. This strategy of ex vivo expansion of IPCs using BBHS provides an autologous therapeutic cell source for the treatment of diabetes. PMID:22310720

  8. Role of Microfluidics in Blood-Brain Barrier Permeability Cell Culture Modeling: Relevance to CNS Disorders.

    Rusanov, Alexander L; Luzgina, Natalia G; Barreto, George E; Aliev, Gjumrakch


    In vitro modeling of the human blood-brain barrier (BBB) is critical for pre-clinical evaluation and predicting the permeability of newly developed potentially neurotoxic and neurotrophic drugs. Here we summarize the specific structural and functional features of endothelial cells as a key component of the BBB and compare analysis of different cell culture models in reflecting these features. Particular attention is paid to cellular models of the BBB in microfluidic devices capable of circulating nutrient media to simulate the blood flow of the brain. In these conditions, it is possible to reproduce a number of factors affecting endothelial cells under physiological conditions, including shear stress. In comparison with static cell models, concentration gradients, which determine the velocity of transport of substances, reproduce more accurately conditions of nutrient medium flow, since they eliminate the accumulation of substances near the basal membrane of cells, not typical for the situation in vivo. Co-cultivation of different types of cells forming the BBB, in separate cell chambers connected by microchannels, allows to evaluate the mutual influences of cells under normal conditions and when exposed to the test substance. New experimental possibilities that can be achieved through modeling of BBB in microfluidic devices determine the feasibility of their use in the practice for pre-clinical studies of novel drugs against neurodegenerative diseases. PMID:26831260

  9. Differential Gene Expression of Primary Cultured Lymphatic and Blood Vascular Endothelial Cells

    Gregory M. Nelson


    Full Text Available Blood vascular endothelial cells (BECs and the developmentally related lymphatic endothelial cells (LECs create complementary, yet distinct vascular networks. Each endothelial cell type interacts with flowing fluid and circulating cells, yet each vascular system has evolved specialized gene expression programs and thus both cell types display different phenotypes. BECs and LECs express distinct genes that are unique to their specific vascular microenvironment. Tumors also take advantage of the molecules that are expressed in these vascular systems to enhance their metastatic potential. We completed transcriptome analyses on primary cultured LECs and BECs, where each comparative set was isolated from the same individual. Differences were resolved in the expression of several major categories, such as cell adhesion molecules (CAMs, cytokines, cytokine receptors. We have identified new molecules that are associated with BECs (e.g., claudin-9, CXCL11, neurexin-1, neurexin-2, the neuronal growth factor regulator-1 and LECs (e.g., claudin-7, CD58, hyaluronan and proteoglycan link protein 1 (HAPLN1, the poliovirus receptor-related 3 molecule that may lead to novel therapeutic treatments for diseases of lymphatic or blood vessels, including metastasis of cancer to lymph nodes or distant organs.

  10. Use of a multiplex PCR to detect and identify Mycobacterium avium and M. intracellulare in blood culture fluids of AIDS patients.

    Kulski, J K; Khinsoe, C; Pryce, T; K. Christiansen


    The presence of mycobacteria in blood culture fluids (BACTEC) of AIDS patients with positive growth indices (GIs, > 20 U) was investigated by using a multiplex PCR to detect and identify members of the genus Mycobacterium, M. avium, M. intracellulare, and M. tuberculosis. Three different methods of extracting mycobacterial DNA from blood culture fluid were compared for use with the multiplex PCR. Mycobacterial cells were pelleted from a small aliquot of blood culture fluid by centrifugation, ...

  11. Gamma-interferon bioassay for detection of bovine tuberculosis in cattle: kinetics of production and dose response in whole blood culture

    Stimulation with mycobacterium bovis PPD sensitised lymphocytes (whole blood or peripheral blood lymphocytes) results in release of gamma-interferon that can be detected by simple bioassay. The optimum concentration of bovine PPD was 20 μg ml and the optimum incubation period was 24 hr for maximum production of gamma-interferon in whole blood culture (128 units/ml) and peripheral blood culture (64 units/ml). (author)

  12. From Blogs to Bottle Caps

    Edinger, Ted


    There is a wonderful community of art educators connecting a once-isolated profession through blogging. Art educators around the world are sharing ideas and communicating with their peers through this amazing resource. In this article, the author describes the bottle cap mural at Tulip Grove Elementary School which was inspired by this exchange of…

  13. Clastogenic interactions of #betta# radiation and caffeine in human peripheral blood cultures

    In order to determine whether the micronucleus test could be used as a rapid assay for mutagenic interactions, we studied the effect of 50-800 R of #betta# radiation in combination with 10-6-10-3 M caffeine in cultured human lymphocytes, with two treatment protocols. In one protocol (T0), whole blood was irradiated with 50-800 R of #betta# radiation, then stimulated with PHA and cultured for 72, 96 or 120 h in the presence or absence of caffeine. Under these conditions, #betta# radiation produced micronuclei in proportion to dose but post-treatment with 1 mM caffein significantly decreased the number of micronuclei observed. The effect of caffeine was greater with the higher radiation doses and at earlier fixation times. Caffeine also decreased the mitotic index which, in turn, decreased the number of micronuclei observed; but caffeine post-treatment still had a significant effect even after mitotic activity was taken into account. In a second protocol (T48), PHA-stimulated (actively cycling) cultures were irradiated 48 h after innoculation, then treated with caffeine, and fixed at 72 h post-innoculation (PI). With this protocol #betta# radiation produced more micronuclei than at T0; this suggests that many of the cells damaged at T0 are either lost or repaired. At T48 1 mM caffeine significantly increased the number of micronuclei observed after #betta# radiation at all doses except 50 and 200 R. The mitotic index increased after 400-600 R, but only in the absence of caffeine. (orig./AJ)

  14. Cytokine profiles in peripheral blood and whole blood cell cultures associated with aggressive periodontitis, juvenile idiopathic arthritis, and rheumatoid arthritis

    Poulsen, Anne Havemose; Sørensen, Lars Korsbaek; Stoltze, Kaj; Bendtzen, Klaus; Holmstrup, Palle


    Cytokines play a key role in the pathogenesis of inflammatory diseases. An obvious question is whether patients with aggressive periodontitis, juvenile idiopathic arthritis, or rheumatoid arthritis share blood cytokine profiles distinguishing them from individuals free of disease....

  15. Qualitative Assessment Of Bottled Water In The Middle East

    Ahmad, S; Saleem, Abdul


    Many brands of bottled water are being produced in the Middle East including the Gulf Cooperation Council (GCC) countries. Over fourteen brands of bottled water could be found in the market of Doha. Use of bottled water has kept on increasing in this region. Reasons for the increase in use of bottled water for drinking have been discussed. The raw water source for the bottled water is groundwater. Most of the manufacturers of the bottled water claim bottled water as "Natural Mineral Water...

  16. Multiplex real-time PCR and blood culture for identification of bloodstream pathogens in patients with suspected sepsis

    Westh, H; Lisby, G; Breysse, F;


    Severe sepsis is increasingly a cause of death. Rapid and correct initial antimicrobial treatment reduces mortality. The aetiological agent(s) cannot always be found in blood cultures (BCs). A novel multiplex PCR test (SeptiFast (alpha version)) that allows identification of 20 bacterial and fung...

  17. Recovery of a catalase-negative Staphylococcus epidermidis strain in blood and urine cultures from a patient with pyelonephritis.

    Kallstrom, George; Chang, Tom; Albertson, Marc; Morilla, Daniel; Fisher, Mark A; Eberly, Bardwell


    This report describes a 60-year-old patient with bilateral nephrolithiasis. A catalase-negative Staphylococcus epidermidis strain was recovered from both urine and blood cultures. Although rare, isolates of catalase-negative Staphylococcus spp., including Staphylococcus aureus, have been reported. Here, we describe the first report of a catalase-negative S. epidermidis strain. PMID:21900516

  18. Recovery of a Catalase-Negative Staphylococcus epidermidis Strain in Blood and Urine Cultures from a Patient with Pyelonephritis ▿

    Kallstrom, George; Chang, Tom; Albertson, Marc; Morilla, Daniel; Fisher, Mark A.; Eberly, Bardwell


    This report describes a 60-year-old patient with bilateral nephrolithiasis. A catalase-negative Staphylococcus epidermidis strain was recovered from both urine and blood cultures. Although rare, isolates of catalase-negative Staphylococcus spp., including Staphylococcus aureus, have been reported. Here, we describe the first report of a catalase-negative S. epidermidis strain. PMID:21900516

  19. SNP may modify the effect of vitamin A supplementation at birth on cytokine production in a whole blood culture assay

    Jørgensen, Mathias Jul; Fisker, Ane Bærent; Erikstrup, Christian;


    Within a neonatal vitamin A supplementation (VAS) trial, we investigated the effect of VAS on TNF-a, IL-10, IL-5 and IL-13 production after lipopolysaccharide, purified protein derivative (PPD) of Mycobacterium tuberculosis and phytohaemagglutinin stimulation using a whole blood culture protocol....

  20. 16S rRNA gene sequencing in routine identification of anaerobic bacteria isolated from blood cultures

    Justesen, Ulrik Stenz; Skov, Marianne Nielsine; Knudsen, Elisa;


    A comparison between conventional identification and 16S rRNA gene sequencing of anaerobic bacteria isolated from blood cultures in a routine setting was performed (n = 127). With sequencing, 89% were identified to the species level, versus 52% with conventional identification. The times for...... identification were 1.5 days and 2.8 days, respectively....

  1. Fluoride and bacterial content of bottled drinking water versus municipal tap water

    Mythri H


    Full Text Available Background: Water is a divine gift. People quench their thirst without questioning the source of water. But, apprehension about contaminants in municipal water supplies along with increased fear of fluorosis made bottled drinking water as one of the important tradable commodities. Objectives: The objectives of the study were to determine and compare the fluoride and bacterial contents of commercially available bottled drinking water and municipal tap water in Davangere city, Karnataka. Materials and Methods: Fifty samples of 10 categories of bottled drinking water with different batch numbers were purchased and municipal water from different sources were collected. Fluoride levels were determined by an ion-selective electrode. Water was cultured quantitatively and levels of bacteria were calculated as colony-forming units (CFUs per milliliter. Results: Descriptive analysis of water samples for fluoride concentration was in the range of 0.07-0.33 for bottled drinking water, Bisleri showing the highest of 0.33. A comparison of the mean values of microbial count for bottled drinking water with that of municipal tap water showed no statistically significant difference, but was more than the standard levels along with the presence of fungus and maggots. Conclusion: The fluoride concentration was below the optimal level for both municipal tap water and bottled drinking water. CFUs were more than the recommended level in both municipal tap water and bottled drinking water.

  2. Culture of normal human blood cells in a diffusion chamber system II. Lymphocyte and plasma cell kinetics

    Normal human blood leukocytes were cultured in Millipore diffusion chambers implanted into the peritoneal cavities of irradiated mice. The evaluation of survival and proliferation kinetics of cells in lymphyocytic series suggested that the lymphoid cells are formed from transition of small and/or large lymphocytes, and the lymphoblasts from the lymphoid cells. There was also evidence indicating that some of the cells in these two compartments are formed by proliferation. The evaluation of plasmacytic series suggested that the plasma cells are formed from plasmacytoid-lymphocytes by transition, and the latter from the transition of lymphocytes. In addition, relatively a small fraction of cells in these two compartments are formed by proliferation. mature plasma cells do not and immature plasma cells do proliferate. Estimation of magnitude of plasma cells formed in the cultures at day 18 indicated that at least one plasma cell is formed for every 6 normal human blood lymphocytes introduced into the culture


    V. N. Zorina


    Full Text Available We had investigated levels of TTG, T4, TNFα, IL-6, IFNγ, and α2-MG in blood serum and supernates of short-term blood cultures in the patients with verified Graves disease before treatment and after reaching of euthyroid status, as compared with healthy controls. We have revealed that initial blood concentrations of free Т4 in the patients were increased, along with decrease in TSH, higher IL-6, IFNγ levels, as well as concentrations of α2-MG which participates in cytokine transport and synthesis. Thiamazole treatment normalized the hormonal profile and reduced blood levels of IL-6, IFNγ and α2-MG, however, without complete normalization, along with increase of serum TNFα contents. It was shown, that the patients before treatment had decreased in vitro response of cells to the mitogenic stimulation as shown by decreased induction of TNFα and IFNγ production, along with, increased spontaneous IFNγ levels. When reaching euthyroid state after Thiamazole administration, we observed an increased spontaneous IFNγ synthesis, decreased IL-6 production in resting cultures. In mitogen-stimulated cell cultures from the treated patients, IFNγ contents became normal, however, TNFα secretion remained lower than in controls. The α2-MG levels in supernates were stable and significantly lower, than in serum. We may presume that thyrotoxicosis treatment with Thiamazole causes stabilization of the endocrine state, however, being not sufficient for normalized production of cytokines, as well as α2-MG, with its regulatory and transporter functions, thus promoting recurrence of disease and reactivation of autoimmune events. 

  4. Diversity of bacteria cultured from the blood of lesser electric rays caught in the northern gulf of Mexico.

    Tao, Zhen; Bullard, Stephen A; Arias, Cova R


    The prevalence and taxonomic diversity of bacteria cultured from the blood of apparently healthy Lesser Electric Rays Narcine bancroftii captured from open beach habitat in the north-central Gulf of Mexico are reported herein. The blood of 9 out of 10 Lesser Electric Rays was positive for bacteria, and bacterial isolates (n = 83) were identified by 16S rRNA gene sequencing. The majority of the isolates belonged to the phylum Proteobacteria (91.5%). Vibrio spp. comprised 53% of all isolates and were recovered from all Lesser Electric Rays with culture-positive blood. Among them, V. harveyi (n = 14) and V. campbellii (n = 11) were most common, followed by a group of unidentified Vibrio sp. (n = 10) related to V. nigripulchritudo. Isolates representing other species of Proteobacteria included Pseudoalteromonas (n = 13), Shewanella (n = 5), Amphritea (n = 3), Nautella (n = 3), and Arenibacter (n = 1). Higher bacterial diversity was observed in blood cultured on marine agar relative to blood agar, but gram-positive bacteria were isolated from the latter only. The 16S rRNA gene sequences of bacterial isolates were compared phylogenetically to those from related type strains. Most isolates were identified to the level of species, but some clustered independently from reference strains, likely representing new species of Vibrio, Amphritea, Shewanella, and Tenacibaculum. The present study is the first record of any bacterium from this ray species and reveals a taxonomically and phylogenetically diverse microbiota associated with its blood. Moreover, these data document that the presence of bacteria in elasmobranch blood is not coincident with clinical signs of disease, thereby rejecting the paradigm of septicemia indicating a disease condition in aquatic vertebrates. PMID:25321403

  5. Mechanism of the clinical effects of uv-irradiated blood: stimulation of dna synthesis by human cells in culture

    This paper studies the DNA-synthetic activity of hyman embryonic cells (EC) cultured in the presence of supernatants from intact and irradiated cell fractions of blood or plasma. Human EC obtained from abortion material were incubated; after incubation, tritium-thymidine was added to the growth medium for 30 min. It is shown that stimulation of DNA synthesis in EC growing in the presence of supernatants from irradiated whole blood is not connected with photoactivation of growth factors in the blood plasma, but takes place as a result of their release from the cells. Donated blood, irradiated with UV light of the same wavelength and within the same dose range as are used under clinical conditions (up to 1200 J/m2), possesses growth-stimulating properties

  6. Determination of growth value thresholds for BACTEC PLUS aerobic blood culture vials.

    McGowan, J E; Metchock, B G


    Growth value thresholds used to identify positive blood culture vials can be defined by users for each BACTEC NR-660 bacteremia detection instrument. Growth values were compared with the recovery of organisms from vials flagged as positive during the testing of 3.056 high-volume vials containing aerobic (BACTEC PLUS 26) medium over a 2-month period. Results showed that optimal threshold values for our use of these vials varied from those recommended by the manufacturer; if the thresholds defined from these data had been used during the study period, total vials flagged as positive from which no organisms were recovered (false alarms) would have been reduced from 181 (5.9/100 vials tested) to 71 (2.3/100 vials tested), with a minimal decrease in the identification of vials containing usual or occasional pathogens (hits). Adjustments of growth value thresholds by the individual user can make the use of BACTEC instruments more efficient by decreasing further processing of vials from which no organisms are recovered. PMID:1572964

  7. Revisiting the IFN-γ release assay: Whole blood or PBMC cultures? - And other factors of influence.

    Hartmann, Sofie Bruun; Emnéus, Jenny; Wolff, Anders; Jungersen, Gregers


    The interferon-γ release assay (IGRA) is a widely used test for the presence of a cell-mediated immune (CMI) response in vitro. This measure is used to test for infection with intracellular pathogens or for validating vaccine efficacy, and it is a widely used test for both human as well as cattle. However, there is no consensus whether to use whole blood cultures or purified PBMCs for the assay, and both cell populations are being used and results compared. Therefore the aim of this study was to compare different culture settings using immune cells from previously vaccinated calves, and to shed light on external factors that could influence the read out in terms of IFN-γ levels. It was found that optimal culture conditions varied between individual animals; when polyclonal activated, cells from whole blood cultures were most responsive, but when activated specifically, the optimal cell concentration/population varied with whole blood, 10×10(6)cells/ml PBMC and 5×10(6)cells/ml PBMC being the highest performing conditions. A further investigation of the distribution of cell populations in PBMCs compared to whole blood was conducted, and a significant (psecreted IFN-γ in whole blood cultures from five calves. Six plates (a-f) were tested and no significant difference in absolute levels of IFN-γ was detected in the six plates when cells were polyclonal and specifically activated. However, we observed a significant (pexpression on plate b, and the relative-to-maximum level on this plate was significant (p<0.05) compared to plate a. Altogether these findings highlight the potential weaknesses of the IFN-γ release assay in terms of the many variables that can influence the results, including the cell culture population, the concentration of cells being cultured, and the plastic ware used for the in vitro culture. These findings stress the importance of documenting the precise assay conditions when publishing results of in vitro IFN-γ release assays. PMID:27073172


    Davidson, P.H.


    Improved mass separator charge bottles are described for containing a dense charge of a chemical compound of copper, nickel, lead or other useful substance which is to be vaporized, and to the method of utilizing such improvcd charge bottles so that the chemical compound is vaporized from the under surface of the charge and thus permits the non-volatile portion thereof to fall to the bottom of the charge bottle where it does not form an obstacle to further evaporation. The charge bottle comprises a vertically disposed cylindrical portion, an inner re-entrant cylindrical portion extending axially and downwardly into the same from the upper end thereof, and evaporative source material in the form of a chemical compound compacted within the upper annular pontion of the charge bottle formed by the re-entrant cylindrical portion, whereby vapor from the chemical compound will pass outwardly from the charge bottle through an apertured closure.

  9. Characterization of feline whole-blood cultures and determination of the frequency of radiation-induced dicentrics in human and feline lymphocytes

    The Ham's F-10 and PHA culture system was applied to whole feline blood, and cell-cycle characteristics such as DNA synthesis and mitotic indices were studied. The results are comparable to those obtained from human whole-blood cultures. The yields of dicentrics were also determined in lymphocytes from X-irradiated human and feline blood. The ratio between the experimental yields of dicentrics in human as compared to feline lymphocytes was 1:0.27. (author)

  10. Bottling process and closure choice influence oxygen levels in wine and wine post-bottling development

    Dimkou, Evdokia


    In the framework of several bottling trials replicating typical winery conditions, Riesling wines were bottled with different levels of dissolved and headspace oxygen and were sealed with co-extruded or screw cap closures in order to investigate the impact of oxygen at bottling as well as oxygen´s ingress through the closure on wine development. Using the luminescence technology, dissolved and headspace oxygen, as well as closure´s oxygen transfer rate were monitored during bottle storage and...

  11. Recent emergence of Staphylococcus aureus clonal complex 398 in human blood cultures.

    Erwin Verkade

    Full Text Available BACKGROUND: Recently, a clone of MRSA with clonal complex 398 (CC398 has emerged that is related to an extensive reservoir in animals, especially pigs and veal calves. It has been reported previously that methicillin-susceptible variants of CC398 circulate among humans at low frequency, and these have been isolated in a few cases of bloodstream infections (BSI. The purpose of this study was to determine the prevalence of S. aureus CC398 in blood cultures taken from patients in a geographic area with a high density of pigs. METHODOLOGY/PRINCIPAL FINDINGS: In total, 612 consecutive episodes of S. aureus BSI diagnosed before and during the emergence of CC398 were included. Three strains (2 MSSA and 1 MRSA that were isolated from bacteremic patients between 2010-2011 were positive in a CC398 specific PCR. There was a marked increase in prevalence of S. aureus CC398 BSI isolated between 2010-2011 compared to the combined collections that were isolated between 1996-1998 and 2002-2005 (3/157, 1.9% vs. 0/455, 0.0%; p = 0.017. CONCLUSIONS/SIGNIFICANCE: In conclusion, in an area with a relative high density of pigs, S. aureus CC398 was found as a cause of BSI in humans only recently. This indicates that S. aureus CC398 is able to cause invasive infections in humans and that the prevalence is rising. Careful monitoring of the evolution and epidemiology of S. aureus CC398 in animals and humans is therefore important.

  12. Inflammatory cytokine detection in adenotonsill and peripheral blood mononuclear cells- culture in adenotonsillectomy patients: a comparative study

    Farhadi M


    Full Text Available Background: Tonsils and adenoid hypertrophy is a major respiratory symptom in children which is partly due to recruitment of inflammatory cells in upper airway lymph nodes as a result of the effects of synthesis and release of different inflammatory cytokines. It seems that infections play role in concert with these cytokines leading to tonsilar hypertrophy and other pathologic consequences. It is proposed that cellular infiltrate of tonsils and adenoids may secrete different quantities of these cytokines compared with peripheral blood mononuclear cells (PBMC cultures.Methods: Among patients who were admitted for adenotonsillectomy to the ENT ward, 37 patients, under 1-12 years old patients with fulfill criteria selected to include the study. Excised adenoid and tonsils cultured and inflammatory cytokines Interferon-γ (INF-γ, Interlukine-1 (IL-1, IL-6, IL-8 and tumor necrosis factor-α (TNF-α measured in cellular culture supernatant. The same cytokines measured in PBMC cultures.Results: The data shows that there is a significant difference between IFN-γ and IL-8 amounts in adenoid tissue culture supernatant and PBMC culture of our patients. Furth-ermore, the amounts of IFN-γ, IL-1 and IL-8 showed considerable difference between tonsilar tissue culture supernatant and PBMC culture of these patients. Although there is a significant correlation between IL-6 amounts in tissue culture supernatant and PBMC culture (P=0.02, the respective data for TNF is only almost significant.Conclusion: Inflammatory cytokines may have significant role in the early provoke of inflammation occurred in hypertrophied tonsils and adenoid. The majority of these cyt-okines increase the expression of adhesion molecules on epithelial cells and influence the recruitment of leucocytes and inflamed tonsils. On the other hand lack of sufficient cytokine release may lead to persistent infections and may cause chronic inflammation and hypertrophied tissue.

  13. Preparation of mycobacterial DNA from blood culture fluids by simple alkali wash and heat lysis method for PCR detection.

    Kulski, J K; Pryce, T


    A sodium iodide-isopropanol (NI) method was compared with an alkali wash and heat lysis (AH) procedure for the preparation and extraction of DNA from BACTEC 13A blood culture fluid samples from AIDS patients for use in a PCR for the detection and identification of mycobacteria. The sensitivity and efficiency of the DNA extraction methods were assessed by a multiplex PCR which detected the members of the genus Mycobacterium and differentiated between M. intracellulare, M. tuberculosis, and M. ...

  14. Comparison of Bactec 9240 and Difco ESP blood culture systems for detection of organisms from vials whose entry was delayed.

    Chapin, K.; Lauderdale, T L


    A comparison of the Bactec 9240 (Becton-Dickinson, Sparks, Md.) and Difco ESP (Difco Laboratories, Detroit, Mich.) instruments for the detection of organism growth from vials whose entry was delayed was evaluated. The instruments' capabilities for organism recovery, time to detection, rates of false-positive results, and numbers of vials in which growth was not detected were made by using seeded blood culture vial pairs and controls with and without delayed entry. Bactec 9240 and Difco ESP ae...

  15. Quantitation of postantibiotic effect by measuring CO2 generation of bacteria with the BACTEC blood culture system.

    Gottfredsson, M; Erlendsdottir, H; Gudmundsson, S.


    The duration of the postantibiotic effect (PAE) determined by bacterial CO2 production measured by using the BACTEC NR 730 blood culture system was compared with PAEs determined by standard viability counting. PAEs for Staphylococcus aureus after exposure to dicloxacillin, vancomycin, rifampin, gentamicin, and ciprofloxacin and for Escherichia coli after exposure to ampicillin, gentamicin, and ciprofloxacin were quantitated by the two methods, and an excellent correlation (r = 0.93) was demon...

  16. Cell differentiation mediated by co-culture of human umbilical cord blood stem cells with murine hepatic cells.

    Stecklum, Maria; Wulf-Goldenberg, Annika; Purfürst, Bettina; Siegert, Antje; Keil, Marlen; Eckert, Klaus; Fichtner, Iduna


    In the present study, purified human cord blood stem cells were co-cultivated with murine hepatic alpha mouse liver 12 (AML12) cells to compare the effect on endodermal stem cell differentiation by either direct cell-cell interaction or by soluble factors in conditioned hepatic cell medium. With that approach, we want to mimic in vitro the situation of preclinical transplantation experiments using human cells in mice. Cord blood stem cells, cultivated with hepatic conditioned medium, showed a low endodermal differentiation but an increased connexin 32 (Cx32) and Cx43, and cytokeratin 8 (CK8) and CK19 expression was monitored by reverse transcription polymerase chain reaction (RT-PCR). Microarray profiling indicated that in cultivated cord blood cells, 604 genes were upregulated 2-fold, with the highest expression for epithelial CK19 and epithelial cadherin (E-cadherin). On ultrastructural level, there were no major changes in the cellular morphology, except a higher presence of phago(ly)some-like structures observed. Direct co-culture of AML12 cells with cord blood cells led to less incisive differentiation with increased sex-determining region Y-box 17 (SOX17), Cx32 and Cx43, as well as epithelial CK8 and CK19 expressions. On ultrastructural level, tight cell contacts along the plasma membranes were revealed. FACS analysis in co-cultivated cells quantified dye exchange on low level, as also proved by time relapse video-imaging of labelled cells. Modulators of gap junction formation influenced dye transfer between the co-cultured cells, whereby retinoic acid increased and 3-heptanol reduced the dye transfer. The study indicated that the cell-co-cultured model of human umbilical cord blood cells and murine AML12 cells may be a suitable approach to study some aspects of endodermal/hepatic cell differentiation induction. PMID:25270685

  17. Development of Syringe/Bottle Hybrids for Sampling Slurries

    A convenient and effective sample bottle system based on simple modifications of disposable plastic syringes and bottles has been devised and tested for slurry samples. Syringe/ bottle hybrids (hereafter referred to as syringe bottles) have the convenience of regular flat-bottom bottles with screw cap closures. In addition, the syringe imparts a sliding and adjustable bottom to the bottle that forces the entire contents from the bottle. The system was designed especially to collect samples for high temperature work-ups of DWPF slurry samples. The syringe bottles together with fixed-bottom sample vial inserts would provide the DWPF with convenient and reliable methods for dealing with slurry samples

  18. Growth capacity of thermotolerant campylobacters in culture media supplemented with pig and cow blood

    Álvaro Tresierra-Ayala; Manuel Navas; Josué Flores; Ramsés Perea; Juan Huanaquiri; María Bendayán; Heriberto Fernández


    In this work 60 thermotolerant Campylobacter strains (37 C. jejuni and 23 C. coli) isolated from the cows, pigs, chickens and ducks (15 strains of each type of animal) were used to establish their growth capacity on media containing cow or swine blood as potential substitutes of sheep or horse blood. The growth capacity was assessed by viable counts on cow and swine blood media, using the modified Miles and Misra method. Campylobacter strains showed better growth in the media supplemented wit...

  19. Interfacial properties of bottle-brush polyelectrolytes

    Claesson, P. M.; Naderi, A.; Iruthayaraj, J.; Pettersson, T.; Vareikis, A.; Makuska, R.; Iruthayaraj, Joseph

    This article is focused on interfacial properties of bottle brush polyelectrolytes, where side-chains are attached along a polymer backbone. This class of polymer has been much less studied than block copolymers, which is particularly true for bottle brush polyelectrolytes with a high graft density...




    Summary A primary rat choroidal epithelial cell culture system was developed to investigate mechanisms of heavy metal toxicity on the blood-cerebrospinal fluid (CSF) barrier. Epithelial cells were dissociated from choroidal tissue by pronase digestion and cultured in standard DMEM culture media supplemented with 10% fetal bovine serum and 10 ng epithelial growth factor per ml. The procedure yielded 2–5 × 104 cells from pooled plexuses of three to four rats, and a viability of 77–85%. The cultures displayed a dominant polygonal type of epithelial cells, with a population doubling time of 2–3 d. The cultures were of distinct choroidal epithelial origins. For example, immunocytochemical studies using monospecific rabbit anti-rat TTR polyclonal antibody revealed a strong positive stain of transthyretin (TTR), a thyroxine transport protein exclusively produced by the choroidal epithelia. Also, reverse-transcriptase polymerase chain reaction (PCR) confirmed the presence of specific TTR mRNA in the cultures. The cultures were further adapted to grow on a freely permeable membrane sandwiched between two culture chambers. The formation of an impermeable confluent monolayer occurred within 5 d after seeding and was verified by the presence of a steady electrical resistance across the membrane (80 ± 10 ohm per cm2). The epithelial barriers appeared to actively transport [125I]-thyroxine from the basal to apical chamber. These results suggest that this primary cell culture system possesses typical choroidal epithelial characteristics and appears to be a suitable model for in vitro mechanistic investigations of blood–CSF barrier. PMID:9542634

  1. Development of a xeno-free autologous culture system for endothelial progenitor cells derived from human umbilical cord blood.

    Sung-Hwan Moon

    Full Text Available Despite promising preclinical outcomes in animal models, a number of challenges remain for human clinical use. In particular, expanding a large number of endothelial progenitor cells (EPCs in vitro in the absence of animal-derived products is the most critical hurdle remaining to be overcome to ensure the safety and efficiency of human therapy. To develop in vitro culture conditions for EPCs derived from human cord blood (hCB-EPCs, we isolated extracts (UCE and collagen (UC-collagen from umbilical cord tissue to replace their animal-derived counterparts. UC-collagen and UCE efficiently supported the attachment and proliferation of hCB-EPCs in a manner comparable to that of animal-derived collagen in the conventional culture system. Our developed autologous culture system maintained the typical characteristics of hCB-EPCs, as represented by the expression of EPC-associated surface markers. In addition, the therapeutic potential of hCB-EPCs was confirmed when the transplantation of hCB-EPCs cultured in this autologous culture system promoted limb salvage in a mouse model of hindlimb ischemia and was shown to contribute to attenuating muscle degeneration and fibrosis. We suggest that the umbilical cord represents a source for autologous biomaterials for the in vitro culture of hCB-EPCs. The main characteristics and therapeutic potential of hCB-EPCs were not compromised in developed autologous culture system. The absence of animal-derived products in our newly developed in vitro culture removes concerns associated with secondary contamination. Thus, we hope that this culture system accelerates the realization of therapeutic applications of autologous hCB-EPCs for human vascular diseases.

  2. Two Cases of Bacterial Pneumonia in Bottle-nosed Dolphins (Tursiops Gillii) at the Seoul Zoo, Korea

    Kyung-Yeon Eo and Oh-Deog Kwon1*


    Two bacterial pneumonia cases of bottle-nosed dolphins (Tursiops gillii) were found in Seoul Zoo. Firstly, severe fibrous pleuropneumonia was revealed in a 14-year-old male bottle-nosed dolphin on its necropsy. The clinical signs included anorexia, listing posture, and vertical submerging after the initial signs of low activity and avoiding trainers. White blood cell count increased up to 31.80 × 103/µl. Despite aggressive antibiotics therapy of intramuscular injection and nebulization it die...

  3. The fluoride content of bottled drinking waters.

    Toumba, K J; Levy, S; Curzon, M E


    Sales of bottled drinking waters in the United Kingdom have tripled over the last 5 years. The fluoride content of 12 bottled waters purchased from two Leeds supermarkets was determined by both the direct and acid diffusion methods and found to vary from 0.10-0.80 mg/l fluoride (ie ppm fluoride). This article shows that bottled drinking waters contain differing concentrations of fluoride. There is no apparent difference between the direct and acid diffusion methods for the determination of fluoride concentrations of drinking waters. The manufacturers' labelling of fluoride concentrations are mainly inaccurate. Dentists should be aware of the fluoride concentrations of the drinking water of their child patients, be they municipal or bottled drinking water, when prescribing fluoride supplements. Also, some parents are using bottled waters to prepare baby milk formulations which themselves may contain high levels of fluoride and subject their children to the risk of dental fluorosis. PMID:8186036

  4. Variation in sister chromatid exchange frequencies between human and pig whole blood, plasma leukocyte, and mononuclear leukocyte cultures

    Sister chromatid exchange (SCE) induction by ultraviolet (UV) light was studied in both human and pig whole blood cultures (WBC) and plasma leukocyte cultures (PLC). No variation in SCE frequency was observed between pig WBC and PLC in control as well as in treated cells. Conversely, SCE frequencies of human PLC were consistently higher than those of WBC in control and UV-exposed cells. Thus, red blood cells (RBCs) do not influence the sensitivity of lymphocytes to UV LIGHT exposure, and there must be some different culture condition(s) in the inducation of SCEs between human WBC and PLC but not in swine lymphocyte cultures. Since the BrdUrd/lymphocyte ratio of WBC was halved in PLC, the effect of BrdUrd concentration in inducing the SCE baseline frequency of PLC may be ruled out. Neither the cell separation technique nor polymorphonuclear leukocytes had a significant role in the elevated SCE frequency of human PLC or MLC. Experiments where human RBCs were titrated into human PLC showed that the induction of an elevated SCE frequency of PLC was suppressed in a dose-dependent manner by the presence of RBCs in the culture medium. Since the incorporation of pig or human RBCs into human PLC as well as into MLC reduced the SCE frequency to that of WBC, a common component and/or function existing in these cells is suggested. Analysis of different RBC components showed that RBCs, specifically RBC ghosts, release a diffusible but not dialyzable corrective factor into culture medium that is able to reduce the SCE frequencies of PLC

  5. The IRIDICA BAC BSI Assay: Rapid, Sensitive and Culture-Independent Identification of Bacteria and Candida in Blood

    Rothman, Richard E.; Peterson, Stephen; Carroll, Karen C.; Zhang, Sean X.; Avornu, Gideon D.; Rounds, Megan A.; Carolan, Heather E.; Toleno, Donna M.; Moore, David; Hall, Thomas A.; Massire, Christian; Richmond, Gregory S.; Gutierrez, Jose R.; Sampath, Rangarajan; Ecker, David J.; Blyn, Lawrence B.


    Bloodstream infection (BSI) and sepsis are rising in incidence throughout the developed world. The spread of multi-drug resistant organisms presents increasing challenges to treatment. Surviving BSI is dependent on rapid and accurate identification of causal organisms, and timely application of appropriate antibiotics. Current culture-based methods used to detect and identify agents of BSI are often too slow to impact early therapy and may fail to detect relevant organisms in many positive cases. Existing methods for direct molecular detection of microbial DNA in blood are limited in either sensitivity (likely the result of small sample volumes) or in breadth of coverage, often because the PCR primers and probes used target only a few specific pathogens. There is a clear unmet need for a sensitive molecular assay capable of identifying the diverse bacteria and yeast associated with BSI directly from uncultured whole blood samples. We have developed a method of extracting DNA from larger volumes of whole blood (5 ml per sample), amplifying multiple widely conserved bacterial and fungal genes using a mismatch- and background-tolerant PCR chemistry, and identifying hundreds of diverse organisms from the amplified fragments on the basis of species-specific genetic signatures using electrospray ionization mass spectrometry (PCR/ESI-MS). We describe the analytical characteristics of the IRIDICA BAC BSI Assay and compare its pre-clinical performance to current standard-of-care methods in a collection of prospectively collected blood specimens from patients with symptoms of sepsis. The assay generated matching results in 80% of culture-positive cases (86% when common contaminants were excluded from the analysis), and twice the total number of positive detections. The described method is capable of providing organism identifications directly from uncultured blood in less than 8 hours. Disclaimer: The IRIDICA BAC BSI Assay is not available in the United States. PMID:27384540

  6. Molecular screening for Candida orthopsilosis and Candida metapsilosis among Danish Candida parapsilosis group blood culture isolates: proposal of a new RFLP profile for differentiation

    Mirhendi, Hossein; Bruun, Brita; Schønheyder, Henrik Carl;


    Candida orthopsilosis and Candida metapsilosis are recently described species phenotypically indistinguishable from Candida parapsilosis . We evaluated phenotyping and molecular methods for the detection of these species among 79 unique blood culture isolates of the C. parapsilosis group obtained...

  7. Reduced release of intact and cleaved urokinase receptor in stimulated whole-blood cultures from human immunodeficiency virus-1-infected patients

    Ostrowski, S R; Piironen, T; Høyer-Hansen, G;


    The blood levels of the soluble forms of the urokinase receptor (suPAR) are increased in human immunodeficiency virus (HIV)-1-infected patients. This study investigated whether the release of urokinase-type plasminogen activator receptor (uPAR) in whole-blood cultures was affected by HIV infection...... controls, whereas the correlation was weaker to leucocytes and nonexisting to circulating suPAR levels in HIV patients. These findings demonstrate that HIV infection affects stimulated and spontaneous uPAR release in whole-blood cultures. Given that high blood levels of suPAR in HIV patients are linked to....... The release of different uPAR forms in whole-blood cultures incubated 24 h with or without phytohemagglutinin and lipopolysaccharide was analysed in 47 HIV patients and 19 controls. suPAR was measured by enzyme-linked immunosorbent assay (ELISA) (bulk-suPAR) and three different time...

  8. Detection of Extended-Spectrum β-Lactamase and Klebsiella pneumoniae Carbapenemase Genes Directly from Blood Cultures by Use of a Nucleic Acid Microarray

    Fishbain, Joel T.; Sinyavskiy, Oleg; Riederer, Kathleen; Hujer, Andrea M.; Robert A Bonomo


    The growing crisis of multidrug-resistant (MDR) Gram-negative bacteria requires that current technologies permit the rapid detection of extended-spectrum β-lactamase (blaESBL) and Klebsiella pneumoniae carbapenemase (blaKPC) genes. In the present study, we assessed the performance characteristics of a commercially available nucleic acid microarray system for the detection of blaESBL and blaKPC genes directly from positive blood cultures. Using blood cultures (BCs) that contained Gram-negative...

  9. Establishment of long-term monocyte suspension cultures from normal human peripheral blood


    The long-term suspension growth of normal, immature myeloid cells from fresh human cord blood was recently reported and required cells separated on supplemented discontinuous Percoll gradients, growth in media containing hydrocortisone and vitamins D3, and gentle, continuous agitation (13). When normal adult bone marrow (six donors) or blood from Epstein-Barr virus (EBV)-seropositive donors (nine donors) was used as a source of fresh human leukocytes, only short-term proliferation of myeloid ...

  10. 27 CFR 27.208 - Liquor bottles denied entry.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Liquor bottles denied... BUREAU, DEPARTMENT OF THE TREASURY LIQUORS IMPORTATION OF DISTILLED SPIRITS, WINES, AND BEER Requirements for Liquor Bottles § 27.208 Liquor bottles denied entry. Filled liquor bottles, not conforming to...

  11. 27 CFR 26.318 - Liquor bottles denied entry.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Liquor bottles denied... BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LIQUORS AND ARTICLES FROM PUERTO RICO AND THE VIRGIN ISLANDS Requirements for Liquor Bottles § 26.318 Liquor bottles denied entry. Filled liquor bottles not conforming...

  12. 27 CFR 1.91 - Bottled distilled spirits.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bottled distilled spirits..., NONINDUSTRIAL USE OF DISTILLED SPIRITS AND WINE, BULK SALES AND BOTTLING OF DISTILLED SPIRITS Bulk Sales and Bottling of Distilled Spirits Warehouse Receipts § 1.91 Bottled distilled spirits. The provisions of...

  13. Efficient generation of multipotent mesenchymal stem cells from umbilical cord blood in stroma-free liquid culture.

    Rowayda Peters

    Full Text Available BACKGROUND: Haematopoiesis is sustained by haematopoietic (HSC and mesenchymal stem cells (MSC. HSC are the precursors for blood cells, whereas marrow, stroma, bone, cartilage, muscle and connective tissues derive from MSC. The generation of MSC from umbilical cord blood (UCB is possible, but with low and unpredictable success. Here we describe a novel, robust stroma-free dual cell culture system for long-term expansion of primitive UCB-derived MSC. METHODS AND FINDINGS: UCB-derived mononuclear cells (MNC or selected CD34(+ cells were grown in liquid culture in the presence of serum and cytokines. Out of 32 different culture conditions that have been tested for the efficient expansion of HSC, we identified one condition (DMEM, pooled human AB serum, Flt-3 ligand, SCF, MGDF and IL-6; further denoted as D7 which, besides supporting HSC expansion, successfully enabled long-term expansion of stromal/MSC from 8 out of 8 UCB units (5 MNC-derived and 3 CD34(+ selected cells. Expanded MSC displayed a fibroblast-like morphology, expressed several stromal/MSC-related antigens (CD105, CD73, CD29, CD44, CD133 and Nestin but were negative for haematopoietic cell markers (CD45, CD34 and CD14. MSC stemness phenotype and their differentiation capacity in vitro before and after high dilution were preserved throughout long-term culture. Even at passage 24 cells remained Nestin(+, CD133(+ and >95% were positive for CD105, CD73, CD29 and CD44 with the capacity to differentiate into mesodermal lineages. Similarly we show that UCB derived MSC express pluripotency stem cell markers despite differences in cell confluency and culture passages. Further, we generated MSC from peripheral blood (PB MNC of 8 healthy volunteers. In all cases, the resulting MSC expressed MSC-related antigens and showed the capacity to form CFU-F colonies. CONCLUSIONS: This novel stroma-free liquid culture overcomes the existing limitation in obtaining MSC from UCB and PB enabling so far unmet

  14. The assessment of genotoxicity of carbamazepine using cytokinesis-block (CB) micronucleus assay in cultured human blood lymphocytes.

    Celik, Ayla


    The genotoxic effect of CBZ has been investigated in few studies. There is little evidence linking carbamazepine (CBZ) with any genotoxic effects, particularly in vitro micronucleus test using cytogenesis-block technique. In this study, the genotoxicity of the antiepileptic drug, carbamazepine, was tested using cytokinesis-block (CB) micronucleus assay. In vitro analysis was performed in human blood lymphocytes from four healthy persons at five different concentrations of carbamazepine (6, 8, 10, 12, 14 microg/mL). Genotoxic potential and cytotoxic effects of carbamazepine were evaluated by using micronucleus assay and cytokinesis-block proliferation index (CBPI), called the parameter of cytotoxicity in human peripheral blood lymphocyte cultures, respectively. The results of this study indicate that CBZ caused the genotoxic effect under in vitro conditions, except at the dose of 6 microg/mL, and cytotoxic effects of carbamazepine were revealed by a decrease in the cytokinesis-block proliferation index at all the concentrations. PMID:16707330

  15. Normalising the Breast: Early Childhood Services Battling the Bottle and the Breast

    Duncan, Judith; Bartle, Carol


    Normalising practices as a tool for controlling the body and bodily processes have been well-documented using Foucault's theories, including debates around breastfeeding. In this article we explore how the ideas of "normalisation" of the bottle-feeding culture of infants in New Zealand early childhood settings has become the…

  16. Multiplex Real-Time PCR Assay for Rapid Detection of Methicillin-Resistant Staphylococci Directly from Positive Blood Cultures

    Wang, Hye-Young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok; Uh, Young; Lee, Hyeyoung


    Methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of mecA, S. aureus, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for Staphylo...

  17. Erratum: Evaluation of CHROMagar Candida, VITEK2 YST and VITEK® MS for identification of Candida strains isolated from blood cultures.

    Mutlu Sariguzel, Fatma; Berk, Elife; Nedret Koc, Ayse; Sav, Hafize; Aydemir, Gonca


    Erratum Following publication of the original article (Infez Med. volume 23, issue 4, pages 318-322, year 2015) we became aware of the following errors which we wish to correct. These corrections have no impact over the study results, their interpretation or conclusions. Title The correct title is the following: Evaluation of chromagenic agar, VITEK2 YST and VITEK® MS for identification of Candida strains isolated from blood cultures Text In the whole text CHROMOMagar Candida shoul be read as chromogenic agar. PMID:27031906

  18. Research on bottom detection in Intelligent Empty Bottle Inspection System

    Bin Huang; Sile Ma; Yufeng Lv; Hualong Zhang; Chunming Liu; Huajie Wang


    Intelligent empty bottle inspection system is an important inspection equipment of empty bottle before filling beer, and it is a blend of machine vision, precision machine and real-time control. They need to cooperate perfectly to achieve the desired effect. In the design of the empty bottle inspection system, one of the key technologies is the bottle bottom detection which affects the speed and accuracy of the system. It includes positioning and defect recognition of bottle bottom. For the...

  19. Influence of relative blood serum amount in culture medium as well as its previous inactivation with heating on nonspecific radioactive iodine binding in vitro studies

    Nonspecific binding of radioactive iodine with the culture medium is directly proportional to the relative blood serum content for concentrations ranging 1-15%. Preliminary inactivation of the serum with heating at 56 degree C for 30 minutes, which is frequently used in in vitro studies, does not influence significantly radioactive iodine binding with culture medium

  20. GLOBEC NEP Rosette Bottle Data (2002)

    National Oceanic and Atmospheric Administration, Department of Commerce — GLOBEC (GLOBal Ocean ECosystems Dynamics) NEP (Northeast Pacific) Rosette Bottle Data from New Horizon Cruise (NH0207: 1-19 August 2002). Notes: Physical data...

  1. Geochemical atlas of European groundwater : bottled water

    Reimann, Clemens; Birke, Manfred; Demetriades, Alecos; Johnson, Christopher


    Analysis of natural bottled mineral water (usually derived from untreated ground water) can provide a first impression of ground water chemistry at the European scale. For this study, 1785 bottled water samples were purchased from supermarkets, representing 1247 wells/springs/boreholes at 884 locations. These were analysed for 72 parameters by a variety of methods. A very strict quality control programme was followed to ensure results of high standard that are presented as a geochemical atlas...

  2. Characteristics of bacterial and fungal growth in plastic bottled beverages under a consuming condition model.

    Watanabe, Maiko; Ohnishi, Takahiro; Araki, Emiko; Kanda, Takashi; Tomita, Atsuko; Ozawa, Kazuhiro; Goto, Keiichi; Sugiyama, Kanji; Konuma, Hirotaka; Hara-Kudo, Yukiko


    Microbial contamination in unfinished beverages can occur when drinking directly from the bottle. Various microorganisms, including foodborne pathogens, are able to grow in these beverages at room temperature or in a refrigerator. In this study, we elucidated the characteristics of microorganism growth in bottled beverages under consuming condition models. Furthermore, we provide insight into the safety of partially consumed bottled beverages with respect to food hygiene. We inoculated microorganisms, including foodborne pathogens, into various plastic bottled beverages and analysed the dynamic growth of microorganisms as well as bacterial toxin production in the beverages. Eight bottled beverage types were tested in this study, namely green tea, apple juice drink, tomato juice, carbonated drink, sport drink, coffee with milk, isotonic water and mineral water, and in these beverages several microorganism types were used: nine bacteria including three toxin producers, three yeasts, and five moulds. Following inoculation, the bottles were incubated at 35°C for 48 h for bacteria, 25°C for 48 h for yeasts, and 25°C for 28 days for moulds. During the incubation period, the number of bacteria and yeasts and visible changes in mould-growth were determined over time. Our results indicated that combinations of the beverage types and microorganism species correlated with the degree of growth. Regarding factors that affect the growth and toxin-productivity of microorganisms in beverages, it is speculated that the pH, static/shaking culture, temperature, additives, or ingredients, such as carbon dioxide or organic matter (especially of plant origin), may be important for microorganism growth in beverages. Our results suggest that various types of unfinished beverages have microorganism growth and can include food borne pathogens and bacterial toxins. Therefore, our results indicate that in terms of food hygiene it is necessary to consume beverages immediately after opening

  3. Prevalence of extended-spectrum beta-lactamases among Enterobacteriaceae isolated from blood culture in a tertiary care hospital

    To determine the prevalence of extended spectrum beta-lactamase among Enterobacteriaceae isolated from blood culture in a tertiary care hospital. We carried out this study at the Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia during the period between January 2003 - December 2004. We tested a total of 601 isolates of the family Enterobacteriaceae from blood culture for the prevalence of extended spectrum beta-lactamase (ESBL) production by the standardized disc diffusion method and confirmed by the ESBL E test strips. Ninety-five (15.8%) of the isolates were ESBL producers. Among these, 48.4% were Klebsiella pneumoniae (K. pneumoniae) followed by15.8% of both Escherichia coli (E. coli) and Enterobacter cloacae (Ent. cloacae). Other isolates produced ESBL in low numbers. Klebsiella pneumoniae produced ESBL in significant numbers. Extended spectrum beta-lactamase gram-negative bacilli present significant diagnostic and therapeutic challenges to the management of infections due to these organisms. Microbiology laboratories should start reporting ESBL producing Enterobacteriaceae organism due to their importance in respect to antibiotic therapy and infection control aspects. (author)

  4. Nucleic Acid, Antibody, and Virus Culture Methods to Detect Xenotropic MLV-Related Virus in Human Blood Samples

    M. F. Kearney


    Full Text Available The MLV-related retrovirus, XMRV, was recently identified and reported to be associated with both prostate cancer and chronic fatigue syndrome. At the National Cancer Institute-Frederick, MD (NCI-Frederick, we developed highly sensitive methods to detect XMRV nucleic acids, antibodies, and replication competent virus. Analysis of XMRV-spiked samples and/or specimens from two pigtail macaques experimentally inoculated with 22Rv1 cell-derived XMRV confirmed the ability of the assays used to detect XMRV RNA and DNA, and culture isolatable virus when present, along with XMRV reactive antibody responses. Using these assays, we did not detect evidence of XMRV in blood samples ( or prostate specimens ( from two independent cohorts of patients with prostate cancer. Previous studies detected XMRV in prostate tissues. In the present study, we primarily investigated the levels of XMRV in blood plasma samples collected from patients with prostate cancer. These results demonstrate that while XMRV-related assays developed at the NCI-Frederick can readily measure XMRV nucleic acids, antibodies, and replication competent virus, no evidence of XMRV was found in the blood of patients with prostate cancer.

  5. Signal pathways underlying homocysteine-induced production of MCP-1 and IL-8 in cultured human whole blood

    Xiao-kun ZENG; You-fei GUAN; Daniel G REMICK; Xian WANG


    Aim: To elucidate the mechanisms underlying homocysteine (Hcy)-induced chemokine production. Methods: Human whole blood was pretreated with inhibitors of calmodulin (CaM), protein kinase C (PKC), protein tyrosine kinase(PTK), mitogen-activated protein kinase (MAPK), and NF-κB and activators of PPARγ for 60 min followed by incubation with Hcy 100 μmol/L for 32 h. The levels of mitogen chemokine protein (MCP)-1 and interleukin-8 (IL-8) were determined by enzyme-linked immunosorbant assay (ELISA). Results: Inhibitors of PKC (calphostin C, 50-500 nmol/L and RO-31-8220, 10-100 nmol/L), CaM(W7, 28-280 μmol/L), ERK1/2 MAPK (PD 98059, 2-20 μmol/L), p38 MAPK(SB 203580, 0.6-6 μmol/L), JNK MAPK (curcumin, 2-10 μmol/L), and NF-κB(PDTC, 10-100 nmol/L) markedly reduced Hcy 100 μmol/L-induced production of MCP-1 and IL-8 in human cultured whole blood, but the inhibitors of PTK(genistein, 2.6-26 μmol/L and tyrphostin, 0.5-5 μmol/L) had no obvious effect on MCP-1 and IL-8 production. PPARγ activators (ciglitazone 30 μmol/L and troglitazone 10 μmol/L) depressed the Hcy-induced MCP-1 production but not IL-8 production in the cultured whole blood. Conclusion: Hcy-induced MCP-1 and IL-8 production is mediated by activated signaling pathways such as PKC,CaM, MAPK, and NF-κB. Our results not only provide clues for the signal transduction pathways mediating Hcy-induced chemokine production, but also offer a plausible explanation for a pathogenic role of hyperhomocysteinemia in these diseases.

  6. Evaluation of a simple Theileria annulata culture protocol from experimentally infected bovine whole blood

    Gharbi M.


    Full Text Available We have evaluated a new simple technique using whole blood from experimentally infected cattle for the isolation and cultivation of Theileria annulata. The study was carried out on 20 Holstein-Frisian bovines that had been experimentally infected with a virulent lethal dose of Theileria annulata. This technique has been compared to the classical peripheral blood monocyte isolation with Ficoll carried out on 22 experimentally infected Holstein-Friesian calves. The effectiveness of the reference technique was estimated to 86.4%, whilst the effectiveness of the new technique was 100%. Moreover, this new technique leads to time and money saving estimated to € 3.06 per sample. It decreases the contamination risks by reducing the steps of sample manipulation.

  7. Evaluation of Capilia TB assay for rapid identification of Mycobacterium tuberculosis complex in BACTEC MGIT 960 and BACTEC 9120 blood cultures

    Muchwa Christopher


    Full Text Available Abstract Background Capilia TB is a simple immunochromatographic assay based on the detection of MPB64 antigen specifically secreted by the Mycobacterium tuberculosis complex (MTC. Capilia TB was evaluated for rapid identification of MTC from BACTEC MGIT 960 and BACTEC 9120 systems in Kampala, Uganda. Since most studies have mainly dealt with respiratory samples, the performance of Capilia TB on blood culture samples was also evaluated. Methods One thousand samples from pulmonary and disseminated tuberculosis (TB suspects admitted to the JCRC clinic and the TB wards at Old Mulago hospital in Kampala, Uganda, were cultured in automated BACTEC MGIT 960 and BACTEC 9120 blood culture systems. BACTEC-positive samples were screened for purity by sub-culturing on blood agar plates. Two hundred and fifty three (253 samples with Acid fast bacilli (AFB, 174 BACTEC MGIT 960 and 79 BACTEC 9120 blood cultures were analyzed for presence of MTC using Capilia TB and in-house PCR assays. Results The overall Sensitivity, Specificity, Positive and Negative Predictive values, and Kappa statistic for Capilia TB assay for identification of MTC were 98.4%, 97.6%, 97.7%, 98.4% and 0.96, respectively. Initially, the performance of in-house PCR on BACTEC 9120 blood cultures was poor (Sensitivity, Specificity, PPV, NPV and Kappa statistic of 100%, 29.3%,7%, 100% and 0.04, respectively but improved upon sub-culturing on solid medium (Middlebrook 7H10 to 100%, 95.6%, 98.2%, 100% and 0.98, respectively. In contrast, the Sensitivity and Specificity of Capilia TB assay was 98.4% and 97.9%, respectively, both with BACTEC blood cultures and Middlebrook 7H10 cultured samples, revealing that Capilia was better than in-house PCR for identification of MTC in blood cultures. Additionally, Capilia TB was cheaper than in-house PCR for individual samples ($2.03 vs. $12.59, respectively, and was easier to perform with a shorter turnaround time (20 min vs. 480 min, respectively

  8. Incidence of Bacterial Isolates from Blood Culture in the Neonatal Intensive Care Unit of Tertiary Care Hospital

    Afif Ahmed


    Full Text Available Background - Even with advancement in the care provided for patients in Neonatal Intensive Care Units (NICU and wide spread use of antibiotics, sepsis remains an important cause of high mortality and morbidity. This study was done to determine the Incidence of bacterial isolates. Objective - We aimed to investigate bacterial pathogens causing neonatal sepsis in the neonatal intensive care unit of Hamad Medical Corporation, Doha, Qatar. Materials and methods - Descriptive and retrospective study between August 2006 and June 2008, Neonatal Intensive Care Unit of Hamad Medical Corporation in Doha, Qatar. All neonates with culture-proven sepsis admitted to Neonatal Intensive Care Unit during study period. Results - Out of 2,851 blood culture sent to the laboratory 302 were positive. These cultures were obtained from 176 neonates resulting in sepsis incidence rates of 6.4 cases per 1,000 live births and case-fatality rates of 17%. Gram positive cocci, fungi, and gram negative bacilli made up 66%, 17.8%, and 16.2% of isolates respectively. Conclusion - Gram positive cocci are the major causes of neonatal sepsis in Doha. The high incidence rates of fungal sepsis are associated with increased mortality risk. Good infection control practice together with sensible antibiotic use and on-going surveillance would result in proper neonatal sepsis management, decrease in associated morbidity and mortality.


    Margaret A. Marshall


    In October and November of 1981 thirteen approaches-to-critical were performed on a remote split table machine (RSTM) in the Critical Mass Laboratory of Pacific Northwest Laboratory (PNL) in Richland, Washington using planar arrays of polyethylene bottles filled with plutonium (Pu) nitrate solution. Arrays of up to sixteen bottles were used to measure the critical number of bottles and critical array spacing with a tight fitting Plexiglas® reflector on all sides of the arrays except the top. Some experiments used Plexiglas shells fitted around each bottles to determine the effect of moderation on criticality. Each bottle contained approximately 2.4 L of Pu(NO3)4 solution with a Pu content of 105 g Pu/L and a free acid molarity H+ of 5.1. The plutonium was of low 240Pu (2.9 wt.%) content. These experiments were sponsored by Rockwell Hanford Operations because of the lack of experimental data on the criticality of arrays of bottles of Pu solution such as might be found in storage and handling at the Purex Facility at Hanford. The results of these experiments were used “to provide benchmark data to validate calculational codes used in criticality safety assessments of [the] plant configurations” (Ref. 1). Data for this evaluation were collected from the published report (Ref. 1), the approach to critical logbook, the experimenter’s logbook, and communication with the primary experimenter, B. Michael Durst. Of the 13 experiments preformed 10 were evaluated. One of the experiments was not evaluated because it had been thrown out by the experimenter, one was not evaluated because it was a repeat of another experiment and the third was not evaluated because it reported the critical number of bottles as being greater than 25. Seven of the thirteen evaluated experiments were determined to be acceptable benchmark experiments. A similar experiment using uranyl nitrate was benchmarked as U233-SOL-THERM-014.

  10. Incidence of thrombocytopenia and changes in various platelet parameters, in blood culture positive neonatal sepsis

    Sartaj Bhat; Suhail Naik; Wasim Rafiq; Syed Tariq A


    Abstract Objective: To assess the incidence of thrombocytopenia and changes in various platelet parameters, in culture positive neonatal sepsis. Methods: This was prospective study conducted over a period of one year from December 2009 to November 2010 in neonatal intensive care unit of DDUH Hospital, a tertiary care hospital in Delhi, North India. All babies who were admitted during this period were evaluated prospectively for evidence of sepsis. Results: sepsis was diagnosed in 560 neonates...

  11. In vitro model of the blood-brain barrier established by co-culture of primary cerebral microvascular endothelial and astrocyte cells

    Yan Wang


    Full Text Available Drugs for the treatment and prevention of nervous system diseases must permeate the blood-brain barrier to take effect. In vitro models of the blood-brain barrier are therefore important in the investigation of drug permeation mechanisms. However, to date, no unified method has been described for establishing a blood-brain barrier model. Here, we modified an in vitro model of the blood-brain barrier by seeding brain microvascular endothelial cells and astrocytes from newborn rats on a polyester Transwell cell culture membrane with 0.4-µm pores, and conducted transepithelial electrical resistance measurements, leakage tests and assays for specific blood-brain barrier enzymes. We show that the permeability of our model is as low as that of the blood-brain barrier in vivo. Our model will be a valuable tool in the study of the mechanisms of action of neuroprotective drugs.

  12. Blood / Money

    Strong, Thomas


    Marilyn Strathern has argued that "nature" in Euro-American culture has appeared as constraint; it has figured the givens of existence on which human artifice is seen to construct "society" or "culture."(5) Among those givens is the notion that human beings are naturally individuals. And blood, too, images individuality: "The very thought of blood, individual blood, touches the deepest feelings in man about life and death" ([RIchard Titmuss] 16.) Transfusion medicine, then, draws on a series ...

  13. Cytotoxicity Investigation on Cultured Human Blood Cells Treated with Single-Wall Carbon Nanotubes

    Maria Rosaria Scarfì


    Full Text Available The single-wall carbon nanotubes (SWCNTs are one of the new materials ofemerging technologies. They are becoming increasingly studied for the possibleapplications in electronics, optics and biology. In particular, very promising fields ofapplication are the development of optical biosensors and the intracellular drug delivery.Nevertheless, there is a paucity of information on their toxicological properties and onpotential human health risk. In the present study the SWCNTs were investigated for thepossible induction of toxicity in human blood cells. Cell growth, viability, apoptosis andmetabolic activity were evaluated in proliferating human peripheral blood lymphocytes. Inun-stimulated human leukocytes primary DNA damage was also evaluated. SWCNTsconcentrations ranging from 1 to 50 μg/ml were tested, and treatment duration varied from6 to 72 h, in accordance with the biological target investigated. A statistically significantdecrease in cell growth was found in cells treated with the highest concentrations (25 and50 μg/ml. Such decrease was not associated to cell death or apoptosis, but it wasdemonstrated to be related to a decrease in metabolic activity, as assessed by resazurinassay. Moreover, treatments of 6 h with SWCNTs concentrations of 1, 5 and 10 μg/mlfailed to induce primary DNA damage on the entire human leukocytes population.

  14. 16S rRNA Gene Sequence-Based Identification of Bacteria in Automatically Incubated Blood Culture Materials from Tropical Sub-Saharan Africa.

    Hagen Frickmann

    Full Text Available The quality of microbiological diagnostic procedures depends on pre-analytic conditions. We compared the results of 16S rRNA gene PCR and sequencing from automatically incubated blood culture materials from tropical Ghana with the results of cultural growth after automated incubation.Real-time 16S rRNA gene PCR and subsequent sequencing were applied to 1500 retained blood culture samples of Ghanaian patients admitted to a hospital with an unknown febrile illness after enrichment by automated culture.Out of all 1500 samples, 191 were culture-positive and 98 isolates were considered etiologically relevant. Out of the 191 culture-positive samples, 16S rRNA gene PCR and sequencing led to concordant results in 65 cases at species level and an additional 62 cases at genus level. PCR was positive in further 360 out of 1309 culture-negative samples, sequencing results of which suggested etiologically relevant pathogen detections in 62 instances, detections of uncertain relevance in 50 instances, and DNA contamination due to sample preparation in 248 instances. In two instances, PCR failed to detect contaminants from the skin flora that were culturally detectable. Pre-analytical errors caused many Enterobacteriaceae to be missed by culture.Potentially correctable pre-analytical conditions and not the fastidious nature of the bacteria caused most of the discrepancies. Although 16S rRNA gene PCR and sequencing in addition to culture led to an increase in detections of presumably etiologically relevant blood culture pathogens, the application of this procedure to samples from the tropics was hampered by a high contamination rate. Careful interpretation of diagnostic results is required.

  15. Age and dose related alteration of in vitro mixed lymphocyte culture response of blood lymphocytes from A-bomb survivors

    The responsiveness of peripheral blood lymphocytes to allogenic antigens in mixed lymphocyte culture (MLC) was measured in 139 atomic-bomb survivors. The study revealed a significant decrease in MLC response with increasing dose of previous radiation exposure. This decline was marked in the survivors who were older than 15 at the time of the bomb (ATB). The results suggest a possible relationship between the recovery of T-cell-related function and the thymic function which processes mature T cells for the immune system. Thus it may be that in the advanced age ATB group, the thymus function had started to involute, allowing less recovery of T-cell function compared to young survivors who had adequate processing T-cell activity

  16. Microbial resistance and frequency of extended-spectrum beta-lactamase (ESBL in isolated from blood cultures

    Ruan Carlos Gomes da Silva


    Full Text Available Introduction:The emergence and spread of isolated carriers of extended-spectrum beta-lactamase (ESBL have complicated the treatment of nosocomial infections, since its production is not easily identified by the sensitivity tests, routinely performed in clinical laboratories, leading to difficulties in the hospital control of resistant microorganisms and antibiotics misuse.Objective:The objective of this study was to analyze the resistance profile and the frequency of ESBL in Gram-negative bacteria isolated from blood cultures. A hundred bacterial samples from blood cultures of adult patients were analyzed, which were phenotypically identified by biochemical tests of carbohydrates fermentation and submitted to determination of the resistance profile by disc diffusion test and ESBL screening by disc approximation and disc replacement methods.Results:Among the bacterial samples tested, 30 were identified as Gram-negative bacteria, predominantly by Proteus mirabilis, Pantoea agglomerans, and Escherichia coli. Of these, 73.33% were positive for the detection of ESBL by phenotypic tests, and was found mainly in Pantoea agglomerans, Proteus mirabilis, and Enterobacter cloacae.Conclusion:The increase in the occurrence of ESBL in different Enterobacteriaceae shows the importance of the amplification of detection in other species than Escherichia coli or Klebsiella sp., so that the assistance to the patient is not restrained, since these resistant bacteria cannot be detected by the laboratories. Considering the frequency of ESBL in this study, we highlight the importance of its detection, aiming to its contribution to the development of improvements in the health care policies of hospitals.

  17. Examination of enzymes concentration in the blood of rats with sepsis caused by mixed and pure bacterial cultures

    Stojanović Dragica


    Full Text Available A clinical form of sepsis induced by cecal ligation and puncture (CLP was caused in order to monitor the concentration of enzymes (alanine aminotransferase - ALT, aspartate aminotransferase - AST, lactate dehydrogenase - LDH, amylase and creatine kinase - CK in the rat blood. Experiments were performed on male Wistar rats, weighing on average 215±25 g. The rats were divided into four groups. In the first three groups (n=28 per group, sepsis was induced by pure culture of Escherichia coli (EC or Staphylococcus aureus (SA and mixed culture (MK of caecum, while the fourth group included 20 control rats who underwent an abdominal incision. Blood was taken in time intervals of 12, 24, 72 and 120 hours. During the experimental protocol, we identified significant changes of all monitored enzymes in the serum of infected rats. After a period of 12 hours there was a significant increase in ALT (all rats with sepsis, AST and LDH (rats in the MK group levels, while a decrease was noted in the concentration of amylase (EC, SA. Similarly, 24 hours after the CLP procedure, a significant decrease of amylase (MK and AST (SA was recorded, while serum LDH level varied significantly from elevated (EC, SA to reduced (MC values. Finally, at the time intervals of 72 and 120 hours the concentration of nearly all monitored enzymes has shown a decline, while significance was noted in lowering of ALT (MK, AST (SA, EC and amylase (SA levels. Statistical significance could not be observed in the change of CK levels at any of examined time points. [Projekat Ministarstva nauke Republike Srbije, br. TR 31071 i br. TR 31079

  18. As a Bacterial Culture Medium, Citrated Sheep Blood Agar Is a Practical Alternative to Citrated Human Blood Agar in Laboratories of Developing Countries

    Russell, F. M.; Biribo, S. S. N.; Selvaraj, G.; Oppedisano, F; Warren, S.; Seduadua, A.; Mulholland, E. K.; Carapetis, J.R.


    Human blood agar (HuBA) is widely used in developing countries for the isolation of bacteria from clinical specimens. This study compared citrated sheep blood agar (CSBA) and HuBA with defibrinated horse blood agar and defibrinated sheep blood agar (DSBA) for the isolation and antibiotic susceptibility testing of reference and clinical strains of Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus. Reference and clinical strains of all organisms were diluted in brain h...

  19. Determination of intracellular cytokines IFN-γ and IL-4 in canine T lymphocytes by flow cytometry following whole-blood culture

    Papadogiannakis, Emmanouil I.; Kontos, Vasilios I.; Tamamidou, Maria; Roumeliotou, Anastasia


    This report describes a whole-blood flow cytometric method for the determination of intracellular cytokines IFN-γ and IL-4 in canine T lymphocyte subpopulations. Conjugated anti-cytokine antibodies and commercially available reagents for cell fixation and permeabilization were used. Canine peripheral blood was cultured with a combination of phorbol-12-myristate-13-acetate (PMA) and ionomycin to promote cytokine synthesis in each cell, along with monensin to increase the sensitivity of the met...

  20. The in vitro effects of artificial and natural sweeteners on the immune system using whole blood culture assays.

    Rahiman, F; Pool, E J


    This article investigates the effects of commercially available artificial (aspartame, saccharin, sucralose) and natural sweeteners (brown sugar, white sugar, molasses) on the immune system. Human whole blood cultures were incubated with various sweeteners and stimulated in vitro with either phytohemagglutinin or endotoxin. Harvested supernatants were screened for cytotoxicity and cytokine release. Results showed that none of the artificial or natural sweeteners proved to be cytotoxic, indicating that no cell death was induced in vitro. The natural sweetener, sugar cane molasses (10 ug/mL), enhanced levels of the inflammatory biomarker IL-6 while all artificial sweeteners (10 ug/mL) revealed a suppressive effect on IL-6 secretion (P < 0.001). Exposure of blood cells to sucralose-containing sweeteners under stimulatory conditions reduced levels of the biomarker of humoral immunity, Interleukin-10 (P < 0.001). The cumulative suppression of Interleukin-6 and Interleukin-10 levels induced by sucralose may contribute to the inability in mounting an effective humoral response when posed with an exogenous threat. PMID:24063614

  1. Potential Impact of a Microarray-Based Nucleic Acid Assay for Rapid Detection of Gram-Negative Bacteria and Resistance Markers in Positive Blood Cultures

    Mancini, Nicasio; Infurnari, Laura; Ghidoli, Nadia; Valzano, Grazia; Clementi, Nicola; Burioni, Roberto; Clementi, Massimo


    We evaluated the Verigene Gram-negative blood culture (BC-GN) test, a microarray that detects Gram-negative bacteria and several resistance genes. A total of 102 positive blood cultures were tested, and the BC-GN test correctly identified 97.9% of the isolates within its panel. Resistance genes (CTX-M, KPC, VIM, and OXA genes) were detected in 29.8% of the isolates, with positive predictive values of 95.8% (95% confidence interval [CI], 87.7% to 98.9%) in Enterobacteriaceae and 100% (95% CI, ...

  2. Human Peripheral Blood Mononuclear Cells Cultured in Normal and Hyperglycemic Media in Simulated Microgravity Using NASA Bioreactors

    Lawless, DeSales


    We sought answers to several questions this summer at NASA Johnson Space Center. Initial studies involved the in vitro culture of human peripheral blood mononuclear in cells in different conditioned culture media. Several human cancer clones were similarly studied to determine responses to aberrant glycosylation by the argon laser. The cells were grown at unit gravity in flasks and in simulated microgravity using NASA bioreactors. The cells in each instance were analyzed by flow cytometry. Cell cycle analysis was acquired by staining nuclear DNA with propidium iodide. Responses to the laser stimulation was measured by observing autofluorescence emitted in the green and red spectra after stimulation. Extent of glycosylation correlated with the intensity of the laser stimulated auto-fluorescence. Our particular study was to detect and monitor aberrant glycosylation and its role in etiopathogenesis. Comparisons were made between cells known to be neoplastic and normal cell controls using the same Laser Induced Autofluorescence technique. Studies were begun after extensive literature searches on using the antigen presenting potential of dendritic cells to induce proliferation of antigen specific cytotoxic T-cells. The Sendai virus served as the antigen. Our goal is to generate sufficient numbers of such cells in the simulated microgravity environment for use in autologous transplants of virally infected individuals including those positive for hepatitis and HIV.

  3. Apigenin ameliorates gamma radiation-induced cytogenetic alterations in cultured human blood lymphocytes.

    Begum, Naziya; Prasad, N Rajendra; Kanimozhi, G; Hasan, Annie Q


    The aim of the present study was to assess the protective effect of apigenin, a dietary flavone, against cytogenetic alterations in human peripheral blood lymphocytes (HPBL) induced by Cobalt-60 radiation (3Gy). Results of MTT [3-(4, 5-dimethyl-2-thiaozolyl)-2,5-diphenyl-2H tetrazolium bromide] assay revealed that 37.2μM of apigenin was found to be non-toxic in HPBL. At this dose (37.2μM) of apigenin, the LD(50) radiation dose of HPBL increased from 2.9Gy to 3.4Gy, which resulted in a DMF of 1.17. Apigenin (37.2μM) treatment 1h before irradiation significantly (p<0.05) reduced DNA damage in irradiated HPBL as measured by comet assay (% tail DNA, tail length, tail moment, and olive tail moment). Moreover, apigenin treatment significantly decreased the frequencies of dicentric (DC), acentric fragments (AF), and acentric rings (AR) in irradiated HPBL. Apigenin pretreatment also reduced the radiation-induced CBMN (cytokinesis blocked micronuclei) anomalies such as micronuclei (MNi), nucleoplasmic bridges (NPB) and nuclear buds (NBUD) in HPBL. These results also showed that there was a significant correlation between NPB and DC frequencies and MNi and AF+AR. Treatment with apigenin alone had no significant effect on DNA damage and chromosomal aberrations in HPBL. Thus, the current studies indicate that apigenin protects HPBL from radiation-induced cytogenetic alterations. PMID:22516036

  4. 27 CFR 26.319 - Used liquor bottles.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Used liquor bottles. 26..., DEPARTMENT OF THE TREASURY LIQUORS LIQUORS AND ARTICLES FROM PUERTO RICO AND THE VIRGIN ISLANDS Requirements for Liquor Bottles § 26.319 Used liquor bottles. The appropriate TTB officer may pursuant...

  5. 27 CFR 31.203 - Possession of used liquor bottles.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Possession of used liquor... BUREAU, DEPARTMENT OF THE TREASURY LIQUORS ALCOHOL BEVERAGE DEALERS Reuse and Possession of Used Liquor Bottles § 31.203 Possession of used liquor bottles. The possession of used liquor bottles by any...

  6. 27 CFR 19.633 - Distinctive liquor bottles.


    ... TTB officer if the bottles are found to— (1) Meet the requirements of 27 CFR part 5; (2) Be... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Distinctive liquor bottles..., DEPARTMENT OF THE TREASURY LIQUORS DISTILLED SPIRITS PLANTS Liquor Bottle and Label Requirements...

  7. 27 CFR 31.202 - Possession of refilled liquor bottles.


    ... liquor bottles. 31.202 Section 31.202 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS ALCOHOL BEVERAGE DEALERS Reuse and Possession of Used Liquor Bottles § 31.202 Possession of refilled liquor bottles. No person who sells, or offers for...

  8. 27 CFR 27.209 - Used liquor bottles.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Used liquor bottles. 27..., DEPARTMENT OF THE TREASURY LIQUORS IMPORTATION OF DISTILLED SPIRITS, WINES, AND BEER Requirements for Liquor Bottles § 27.209 Used liquor bottles. The appropriate TTB officer may pursuant to letterhead...

  9. 27 CFR 24.308 - Bottled or packed wine record.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bottled or packed wine record. 24.308 Section 24.308 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS WINE Records and Reports § 24.308 Bottled or packed wine record. A proprietor who bottles, packs,...

  10. 27 CFR 24.255 - Bottling or packing wine.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bottling or packing wine. 24.255 Section 24.255 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS WINE Storage, Treatment and Finishing of Wine Bottling, Packing, and Labeling of Wine § 24.255 Bottling...

  11. 27 CFR 24.256 - Bottle aging wine.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bottle aging wine. 24.256 Section 24.256 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS WINE Storage, Treatment and Finishing of Wine Bottling, Packing, and Labeling of Wine § 24.256 Bottle aging wine....

  12. 21 CFR 1250.42 - Water systems; constant temperature bottles.


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Water systems; constant temperature bottles. 1250...; constant temperature bottles. (a) The water system, whether of the pressure or gravity type, shall be... at all times as to prevent contamination of the water. (e) Constant temperature bottles and...

  13. 27 CFR 19.749 - Bottling and packaging record.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bottling and packaging record. 19.749 Section 19.749 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... Account § 19.749 Bottling and packaging record. The bottling and packaging record shall be prepared...

  14. Detection of Intracellular Adhesion (ica Gene and Biofilm Formation Staphylococcus aureus Isolates from Clinical Blood Cultures

    Mohsen Mirzaee


    Full Text Available Background: In fact the biofilms are composed of bacterial cells living inmulticellular structures such as tissues and organs embedded within a self-produced matrix of extracellular polymeric substance (EPS. Ability to attach and biofilm formation are the most important virulence factors Staphylococcus aureus isolates. The aims of this study were to detect intracellular adhesion (ica locus and its relation to the biofilm formation phenotype in clinical isolates of S. aureus isolated from bloodcultures.Methods: A total of 31 clinical S. aureus isolates were collected from Loghman Hospital of Tehran, Iran. In vitro biofilm formation ability was determined by microliter tissue culture plates. All clinical isolates were examined for determination the ica locus by using PCR method.Results: Twelve (38.7% of the isolates were strong biofilm producers. The results showed that 18(80.6% of the isolates carried icaD gene, whereas the prevalence of icaA, icaB and icaC were 51.6%, 45.1% and 77.4% respectively.Conclusions: S. aureus clinical isolates have different ability to form biofilm. This may be caused by the differences in the expression of biofilm related genes, genetic make-up and physiological conditions.

  15. Cross-Cultural Validation of the High Blood Pressure Health Literacy Scale in a Chinese Community.

    Qinghua Zhang

    Full Text Available Considering the importance of health literacy (HL for the maximum yield from the hypertension control programs, development of a reliable and valid instrument of hypertension-related HL is critical. This study aimed to translate and validate the High Blood Pressure-Health Literacy Scale (HBP-HLS into Chinese (C-HBP-HLS and evaluate its psychometric properties in Chinese context.Between June 2013 and January 2014, a cross-sectional study was conducted among recruited hypertensive patients belonging to the Han and Kazakh-Chinese communities in Urumqi, Xinjiang, China.A pilot sample (n = 242 was selected for the exploratory factor analysis of the translated and modified instrument. Another sample (n = 308 was recruited for the confirmatory factor analysis. C-HBP-HLS consisted of five dimensions (Print Health Literacy, Medication Label, Understanding Ability, Newest Vital Sign Test, and Avoiding Food Allergy containing 15 items, accounting for 77.7% of the total variance. The 5-factor model demonstrated a good overall fit. The scale-level content validity index was 0.85. Cronbach's alpha of the overall scale was 0.78 and test-retest reliability was 0.96. Education level had a strong positive correlation with the scores for items Q1, Q2, and Q3(r = 0.481, 0.492, 0.475, respectively. Health Literacy scores among Kazakh patients were significantly lower than Han (7.13±7.90 vs. 30.10±13.42, Z = -14.573, P<0.001.C-HBP-HLS demonstrated suitable factor structure and robust psychometric properties for measuring health literacy level among hypertensive patients in China.

  16. Cross-Cultural Validation of the High Blood Pressure Health Literacy Scale in a Chinese Community

    Zhang, Qinghua; Huang, Feifei; Liu, Zaoling; Zhang, Na; Mahapatra, Tanmay; Tang, Weiming; Lei, Yang; Dai, Yali; Tang, Songyuan; Zhang, Jingping


    Background Considering the importance of health literacy (HL) for the maximum yield from the hypertension control programs, development of a reliable and valid instrument of hypertension-related HL is critical. This study aimed to translate and validate the High Blood Pressure-Health Literacy Scale (HBP-HLS) into Chinese (C-HBP-HLS) and evaluate its psychometric properties in Chinese context. Method Between June 2013 and January 2014, a cross-sectional study was conducted among recruited hypertensive patients belonging to the Han and Kazakh-Chinese communities in Urumqi, Xinjiang, China. Results A pilot sample (n = 242) was selected for the exploratory factor analysis of the translated and modified instrument. Another sample (n = 308) was recruited for the confirmatory factor analysis. C-HBP-HLS consisted of five dimensions (Print Health Literacy, Medication Label, Understanding Ability, Newest Vital Sign Test, and Avoiding Food Allergy) containing 15 items, accounting for 77.7% of the total variance. The 5-factor model demonstrated a good overall fit. The scale-level content validity index was 0.85. Cronbach’s alpha of the overall scale was 0.78 and test-retest reliability was 0.96. Education level had a strong positive correlation with the scores for items Q1, Q2, and Q3(r = 0.481, 0.492, 0.475, respectively). Health Literacy scores among Kazakh patients were significantly lower than Han (7.13±7.90 vs. 30.10±13.42, Z = -14.573, P<0.001). Conclusion C-HBP-HLS demonstrated suitable factor structure and robust psychometric properties for measuring health literacy level among hypertensive patients in China. PMID:27116336

  17. Smart specialisation - Old wine in new bottles or new wine in old bottles?

    Bjorn Asheim


    The paper deals with the problematic addressed in the above subtitle; i.e. that ?smart specialization? indicates old wine (specialization) in new bottles (smart), but is in fact new wine (diversity) in old bottles (specialization). This confusion will obviously lead to problems with respect to the content, communication and implementation of the strategy, and, thus, potentially reduce its impact. Specializaiton has been the main industrial and regional development strategy since the days of M...

  18. Dr. von Braun Discusses 'Bottle Suit' Concept


    Dr. Wernher von Braun (center), then Chief of the Guided Missile Development Division at Redstone Arsenal, Alabama, discusses a 'bottle suit' model with Dr. Heinz Haber (left), an expert on aviation medicine, and Willey Ley, a science writer on rocketry and space exploration. The three men were at the Disney studios appearing in the motion picture, entitled 'Man in Space.'

  19. 27 CFR 4.26 - Estate bottled.


    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Estate bottled. 4.26 Section 4.26 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Standards of Identity for Wine § 4.26...

  20. 21 CFR 165.110 - Bottled water.


    ... regulations (40 CFR part 143) are as follows: Contaminant Concentration in milligrams per liter Aluminum 0.2....C. 552(a) and 1 CFR part 51, or (7) Method 505—“Analysis of Organohalide Pesticides and Commercial... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Bottled water. 165.110 Section 165.110 Food...

  1. Comparing clinical and microbiological methods for the diagnosis of true bacteraemia among patients with multiple blood cultures positive for coagulase-negative staphylococci.

    Al Wohoush, I; Rivera, J; Cairo, J; Hachem, R; Raad, I


    We assessed the accuracy of the Centers for Disease Control and Prevention (CDC) clinical criteria as well as other microbiological methods for the diagnosis of coagulase-negative staphylococci bacteraemia. The CDC clinical criteria had low accuracy, which can be improved by speciation, particularly if the patient had more than two positive blood cultures. PMID:20854425

  2. Molecular screening for Candida orthopsilosis and Candida metapsilosis among Danish Candida parapsilosis group blood culture isolates: proposal of a new RFLP profile for differentiation

    Mirhendi, Hossein; Bruun, Brita; Schønheyder, Henrik Carl;


    Candida orthopsilosis and Candida metapsilosis are recently described species phenotypically indistinguishable from Candida parapsilosis . We evaluated phenotyping and molecular methods for the detection of these species among 79 unique blood culture isolates of the C. parapsilosis group obtained...... number of invasive infections in Denmark....

  3. Impact of positive chest X-ray findings and blood cultures on adverse outcomes following hospitalized pneumococcal lower respiratory tract infection

    Skovgaard, Marlene; Schønheyder, Henrik C; Benfield, Thomas Lars Vibe;


    Little is known about the clinical presentation and outcome of pneumococcal lower respiratory tract infection (LRTI) without positive chest X-ray findings and blood cultures. We investigated the prognostic impact of a pulmonary infiltrate and bacteraemia on the clinical course of hospitalized...... patients with confirmed pneumococcal LRTI....

  4. Methods for evaluating hermetic closures for screw-capped bottles.

    Cilento, R D


    With the described method, a large number of sealed bottles can be tested to determine their effectiveness, integrity, and reliability in preserving an inert atmosphere. The method consists of placing milligram quantities of dry ice into the bottles, capping them, and periodically weighing them on an analytical balance; leaky bottles are detected by a loss in weight. This method can be applied to test a large number of bottles with a minimum of effort and manpower and without complicated instrumentation. The data presented are highly reproducible and correlate well with data from other methods and with the physical defects of the bottles. PMID:845797

  5. Growth of Scytalidium sp. in a counterfeit bevacizumab bottle

    Gerardo Garcia-Aguirre


    Full Text Available After drawing a dose from an closed bevacizumab (Avastin bottle, a fungus-like foreign body was observed inside. Samples from the vial were cultured in Sabouraud Emmons media. Growth of multiple light brown colonies with dark pigment was observed after 10 days. The species was identified as Scytalidium sp.Vial, analysis reported that the seal was lacking proper identification measures and that the label, batch number and expiry date did not correspond to a genuine product. Chemical analysis showed no protein, but 3% of polyethylene glycol, citrate and ethanol. Counterfeit bevacizumab is a real situation that poses a significant risk for ophthalmology and oncology patients. The medical community should be aware of this situation in order to enforce adequate preventive measures.

  6. Enrichment methodology to increase the positivity of cultures from body fluids

    Alessandra Valle Daur


    Full Text Available Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially-severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1 the conventional culture method (agar plating and (2 the enrichment culture technique, using the Bact/Alert® blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents.

  7. The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002

    Mirović Veljko


    Full Text Available The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002. The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures. The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%, during the follow-up period. None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2-72.2%. In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA fenotype. Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1% during the follow-up period. Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001. Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates. Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time. Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period.

  8. Revisiting the IFN-γ release assay: Whole blood or PBMC cultures? - And other factors of influence

    Hartmann, Sofie Bruun; Emnéus, Jenny; Wolff, Anders;


    The interferon-γ release assay (IGRA) is a widely used test for the presence of a cell-mediated immune (CMI) response in vitro. This measure is used to test for infection with intracellular pathogens or for validating vaccine efficacy, and it is a widely used test for both human as well as cattle...... choice of incubation plate would interfere with the level of secreted IFN-γ in whole blood cultures from five calves. Six plates (a–f) were tested and no significant difference in absolute levels of IFN-γ was detected in the six plates when cells were polyclonal and specifically activated. However, we...... observed a significant (p < 0.05) higher background level in a flat-bottom plate from Corning® (cat# 3595) (plate d) compared to two different flat-bottom plates from Corning® (cat# 3596) (plate b) and Nunc™ (cat# 167008) (plate a). Furthermore 4 out of 5 calves had maximum specific IFN-γ expression on...

  9. Clinical-scale cultures of cord blood CD34(+) cells to amplify committed progenitors and maintain stem cell activity.

    Ivanovic, Zoran; Duchez, Pascale; Chevaleyre, Jean; Vlaski, Marija; Lafarge, Xavier; Dazey, Bernard; Robert-Richard, Elodie; Mazurier, Frédéric; Boiron, Jean-Michel


    We developed a clinical-scale cord blood (CB) cell ex vivo procedure to enable an extensive expansion of committed progenitors--colony-forming cells (CFCs) without impairing very primitive hematopoietic stem cells (HSCs). CD34(++) cells, selected from previously cryopreserved and thawed CB units, were cultured in two steps (diluted 1:4 after 6 days) in the presence of stem cell factor (SCF), fms-related tyrosine kinase 3 ligand (Flt-3L), megakaryocyte growth and development factor (MGDF) (100 ng/ml each), granulocyte-colony stimulating factor (G-CSF) (10 ng/ml) in HP01 serum-free medium. HSC activity was evaluated in a serial transplantation assay, by detection of human cells (CD45, CD33, CD19 and CFC of human origin) in bone marrow (BM) of primary and secondary recipient NOD/SCID mice 6-8 weeks after transplantation. A wide amplification of total cells (∼350-fold), CD34(+) cells (∼100-fold), and CFC (∼130-fold) without impairing the HSC activity was obtained. The activity of a particular HSC subpopulation (SRC(CFC)) was even enhanced.Thus, an extensive ex vivo expansion of CFCs is feasible without impairing the activity of HSCs. This result was enabled by associating antioxidant power of medium with an appropriate cytokine cocktail (i.e., mimicking physiologic effects of a weak oxygenation in hematopoietic environment). PMID:21294956

  10. Lying behaviour and IgG-levels of newborn calves after feeding colostrum via tube and nipple bottle feeding.

    Bonk, Stephanie; Nadalin, Audrey; Heuwieser, Wolfgang; Veira, Douglas


    Oesophageal tube feeding colostrum is used to ensure sufficient colostrum intake in newborn calves but the impact of tube feeding on animal behaviour is unclear. Therefore the objective of this study was to compare lying behaviour of tube-fed or bottle-fed dairy calves. Calves (n = 37) in 3 groups were offered 3·5 l colostrum 2 h after birth. Calves of the bottle group were fed with a nipple bottle. Calves of the placebo tubing group were tubed for 4 min but no colostrum was given and they were then fed with a nipple bottle. Calves of the tubing group received 3·5 l colostrum via tube feeding. Consumed amount of bottle and placebo tubing calves was recorded. If they refused some of the offered 3·5 l the rest was offered in a second feeding 2 h later. Lying behaviour was measured by data loggers fitted to right hind leg for 3 d. Blood samples were taken 24 h after birth for determination of IgG concentration. The voluntary colostrum intake differed significantly between bottle-fed and placebo tubed calves at first feeding. Considering both colostrum feedings, bottle-fed calves consumed 3·44 ± 0·14 l and placebo tubed calves consumed 3·20 ± 0·38 l colostrum. ImmunoglobulinG intake (255·6 ± 77·5 g IgG), serum IgG concentration 24 h after birth (22·8 ± 6·7 g/l) and total serum protein concentration (6·1 ± 0·6 g/dl) did not differ between groups. None of the calves had a failure of passive transfer. There was no effect of tubing on lying behaviour. PMID:27600963